HK40074219A

HK40074219A - 6,7-dihydro-5h-pyrido[2,3-c]pyridazine derivatives and related compounds as bcl-xl protein inhibitors and pro-apoptotic agents for treating cancer

Info

Publication number: HK40074219A
Application number: HK62022063323.9A
Authority: HK
Inventors: Novak Tibor; Edward Paul Davidson James; Paczal Attila; -Benoit Starck Jérôme; Kotschy ANDRÁS; Brooke Murray James; Bedford Simon; Chanrion Maïa; Colland Frédéric; Philip Dodsworth Mark; HERNER András; Leticia Maragno Ana; SANDERS Emma; Pál Timári Mátyás; Webb Paul
Original assignee: Les Laboratoires Servier; Vernalis (R&D) Limited
Priority date: 2019-07-29
Filing date: 2020-07-28
Publication date: 2022-12-30

Description

6, 7-dihydro-5H-pyrido [2,3-C ] pyridazine derivatives and related compounds as BCL-XL protein inhibitors and pro-apoptotic agents for the treatment of cancer

Technical Field

The present invention relates to 6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl derivatives, pharmaceutical compositions containing them and their use as pro-apoptotic agents. The compounds of the invention inhibit the activity of the Bcl-xL protein and may be of interest in the treatment of cancer, immune and autoimmune diseases.

Background

Apoptosis (programmed cell death) is an evolutionarily conserved pathway that is critical for tissue homeostasis, development, and clearance of damaged cells. Dysregulation of apoptosis contributes to human diseases including malignancies, neurodegenerative disorders, immune system diseases and autoimmune diseases (Hanahan and Weinberg, cell.2011 Mar 4; 144(5): 646-74; Marsden and Strasser, Annu Rev Immunol.2003; 21: 71-105; Vaux and Flavell, Curr Opin Immunol.2000 Dec; 12(6): 719-24). Escape of apoptosis is considered a hallmark of cancer, involved in tumor development as well as sustained tumor expansion and resistance to anticancer therapy (Hanahan and Weinberg, cell.2000 Jan 7; 100(1): 57-70).

The Bcl-2 protein family comprises key regulators of cell survival that can either inhibit (e.g., Bcl-2, Bcl-xL, Mcl-1) or promote (e.g., Bad, Bax) apoptosis (Gross et al, Genes Dev.1999 Aug 1; 13(15): 1899-.

Whether a cell survives or undergoes apoptosis in the face of stress stimuli depends on the degree of pairing between the Bcl-2 family member that promotes cell death and the family member that promotes cell survival. To a large extent, these interactions involved the docking of the Bcl-2 homology 3(BH3) domain of a pro-apoptotic family member into a groove on the surface of a pro-survival member. The presence of the Bcl-2 homology (BH) domain defines the members of the Bcl-2 family, which are divided into three major groups based on the specific BH domain present in the protein. Survival-promoting members such as Bcl-2, Bcl-xL and Mcl-1 contain BH domain 1-4, while pro-apoptotic effectors of mitochondrial outer membrane permeabilization, Bax and Bak, contain BH domain 1-3 during apoptosis (Youle and Straser, Nat. Rev. mol. cell biol.2008 Jan; 9(1): 47-59).

Overexpression of viable members of the Bcl-2 family is a hallmark of cancer, and these proteins have been shown to play an important role in tumor development, maintenance, and resistance to anticancer therapies (Czabotar et al, nat. rev. mol. cell biol.2014 Jan; 15(1): 49-63). Bcl-xL (also known as BCL2L1, from BCL2-like 1) is frequently amplified in Cancer (Beroukhim et al, Nature 2010 Feb 18; 463(7283): 899-.

Furthermore, several studies using transgenic knockout mouse models and transgenic overexpression of Bcl-2 family members highlighted the importance of these proteins in immune system and autoimmune diseases (for a review see Merino et al, Apoptosis 2009 Apr; 14(4):570-83.doi:10.1007/s 10495-008-. Transgenic overexpression of Bcl-xL in the T cell compartment resulted in resistance to apoptosis induced by glucocorticoid, g-radiation, and CD3 cross-linking, suggesting that transgenic Bcl-xL overexpression may reduce apoptosis in resting and activated T cells (Droin et al, Biochim Biophys Acta 2004 Mar 1; 1644(2-3):179-88.doi:10.1016/j. bbamcr.2003.10.011.PMID: 14996502). In patient samples, sustained or high expression of anti-apoptotic Bcl-2 family proteins has been observed (Pope et al, Nat Rev Immunol.2002 Jul; 2(7):527-35.doi: 10.1038/nrix.846. PMID: 12094227). In particular, T cells isolated from the joints of rheumatoid arthritis patients exhibit increased expression of Bcl-xL and are resistant to spontaneous apoptosis (Saln et al, J Clin invest.1997 Feb 1; 99(3):439-46.doi:10.1172/JCI119178.PMID: 9022077). The use of BH3 mimetics has also shown benefit in preclinical models of immune system disease and autoimmune disease. Treatment with ABT-737(Bcl-2, Bcl-xL and Bcl-w inhibitors) resulted in effective inhibition of lymphocyte proliferation in vitro. Importantly, mice treated with ABT-737 in animal models of arthritis and lupus show a significant reduction in disease severity (Bardwell et al, J Clin invest.1997 Feb 1; 99(3):439-46.doi:10.1172/JCI119178.PMID: 9022077). Furthermore, ABT-737 has been shown to prevent allogeneic T-cell activation, proliferation and cytotoxicity in vitro, and to inhibit allogeneic T-cell and B-cell responses with high selectivity for lymphoid cells after skin transplantation (Cippa et al, Transpl int.2011 Jul; 24(7):722-32.doi:10.1111/j.1432-2277.2011.01272.x.Epub 2011 May 25.PMID: 21615547).

The above findings have prompted the discovery and development of a new class of drugs known as BH3 mimetics. These molecules are capable of disrupting the interaction between pro-apoptotic and anti-apoptotic members of the Bcl-2 family and are potent inducers of apoptosis. Such novel agents include inhibitors of Bcl-2, Bcl-xL, Bcl-w, and Mcl-1. The first BH3 mimetics described were ABT-737 and ABT-263, which target Bcl-2, Bcl-xL, and Bcl-w (Park et al, j.med.chem.2008 Nov 13; 51(21): 6902-15; Roberts et al, j.clin.oncol.2012 Feb 10; 30(5): 488-96). Thereafter, selective inhibitors of Bcl-2 (ABT-199 and S55746-Souers et al, Nat Med.2013 Feb; 19(2): 202-8; Casara et al, Oncotarget 2018 Apr 13; 9(28):20075-, nat. commun.2018 Dec 17; 9(1):5341). Selective Bcl-2 inhibitor ABT-199 is now approved for the treatment of CLL and AML patients in combination therapy, while other inhibitors are still in preclinical or clinical development. In preclinical models, ABT-263 has shown activity in several hematologic malignancies and solid tumors (Shoemaker et al, Clin. Cancer Res.2008Jun 1; 14(11): 3268-77; Ackler et al, Cancer Chemotherm. Pharmacol.2010 Oct; 66(5): 869-80; Chen et al, mol. Cancer Ther.2011 Dec; 10(12): 2340-9). In clinical studies, ABT-263 showed objective anti-tumor activity in lymphoid malignancies (Wilson et al, Lancet oncol.2010 Dec; 11(12): 1149-59; Roberts et al, j.clin.oncol.2012 Feb 10; 30(5):488-96), and its activity in solid tumors is being studied in combination with several therapies. Selective Bcl-xL inhibitors A-1155463 or A-1331852 showed in vivo activity in T-ALL (T-cell acute lymphoblastic leukemia) and preclinical models of different types of solid tumors (Tao et al, ACS Med Chem Lett.2014 Aug 26; 5(10): 1088-93; Leverson et al, Sci. Transl. Med.2015 Mar 18; 7(279):279ra 40). Mcl-1 selective inhibitors have shown promising in vivo activity in several types of hematological malignancies in preclinical models, and three of them (S64315, AMG176 and AZD5991) are currently being studied in clinical trials (Yang et al, eur.j.med.chem.2019 May 8; 177: 63-75). Thus, BH3 mimetics represent a highly attractive approach to the development of new therapies in the fields of oncology as well as immune and autoimmune diseases. In particular, there is a need for small molecules that selectively inhibit Bcl-xL proteins. The present invention satisfies this need.

Summary of The Invention

The present invention provides potent and selective Bcl-xL inhibitors of formula (I) as defined below. We have demonstrated that these compounds are able to induce apoptosis of cancer cells in vivo, thereby triggering tumor regression in mice. Based on their pro-apoptotic properties, the compounds of the invention are useful in the treatment of pathologies involving dysregulation of apoptosis, such as cancer, autoimmune diseases and diseases of the immune system. Furthermore, these compounds were well tolerated in mice with no clinically relevant weight loss after treatment with effective doses, suggesting that there is a possible therapeutic margin for using these Bcl-xL targeting small molecules in cancer therapy. Consistent with the previously described role of Bcl-xL in regulating platelet longevity (Zhang et al, Cell Death differ.2007 May; 14(5): 943-51; Mason et al, cell.2007 Mar 23; 128(6):1173-86), we observed a reduction in the number of circulating platelets after treatment of mice with these inhibitors, and recovery after treatment cessation. In view of this effect on platelet survival, the Bcl-xL inhibitors of the invention may also be useful in the treatment of diseases or disorders characterized by excessive or deregulated platelet activity, such as pre-thrombotic disorders.

Detailed Description

In a first embodiment (E1), the invention provides a compound of formula (I):

wherein:

het moieties represent fused aromatic or non-aromatic rings consisting of 5 to 7 ring members, which may contain, in addition to nitrogen, an additional heteroatom or group selected from oxygen, sulphur and C ═ O,

◆A₄and A₅Independently of one another, represent a carbon or nitrogen atom,

preferably A₄And A₅All represent a nitrogen atom, and are,

◆Z₁represents a bond, -N (R) -or-O-, wherein R represents hydrogen or a linear or branched C₁-C₆An alkyl group, a carboxyl group,

◆R₁represents a group selected from: hydrogen; optionally substituted by hydroxy or C₁-C₆Alkoxy-substituted straight or branched C₁-C₆An alkyl group; c₃-C₆A cycloalkyl group; a trifluoromethyl group; straight or branched C₁-C₆Alkylene-heterocycloalkyl, in which the heterocycloalkyl is optionally substituted by a straight-chain or branched C₁-C₆Alkyl substitution;

◆R₂represents hydrogen or methyl;

◆R₃represents a group selected from: hydrogen; straight or branched C₁-C₄An alkyl group; -X₁-NR_aR_b；-X₁-N⁺R_aR_bR_c；-X₁-O-R_c；-X₁-COOR_c；-X₁-PO(OH)₂；-X₁-SO₂(OH)；-X₁-N₃And

◆R_aand R_bIndependently of one another, represent a group selected from: hydrogen; a heterocycloalkyl group; -SO₂Phenyl, wherein the phenyl radical may be linear or branched C₁-C₆Alkyl substitution; straight or branched chain C optionally substituted by one or two hydroxy groups₁-C₆An alkyl group; c₁-C₆alkylene-SO₂OH；C₁-C₆alkylene-SO₂O^-；C₁-C₆alkylene-COOH; c₁-C₆alkylene-PO (OH) ₂；C₁-C₆alkylene-NR_dR_e；C₁-C₆alkylene-N⁺R_dR_eR_f；C₁-C₆Alkylene-phenyl, in which the phenyl radical may be substituted by C₁-C₆Alkoxy substitution;

the following groups:

or R_aAnd R_bForm a ring B with the nitrogen atom carrying them₁；

Or R_a、R_bAnd R_cForm a bridge C with the nitrogen atom carrying them₃-C₈A heterocycloalkyl group, a heterocyclic alkyl group,

◆R_c、R_d、R_e、R_findependently of one another, represents hydrogen or straight-chain or branched C₁-C₆An alkyl group, a carboxyl group,

or R_dAnd R_eForm a ring B with the nitrogen atom carrying them₂，

Or R_d、R_eAnd R_fForm a bridge C with the nitrogen atom carrying them₃-C₈A heterocycloalkyl group, a heterocyclic alkyl group,

◆Het₁represents a group selected from:

◆Het₂represents a group selected from:

◆A₁is-NH-, -N (C)₁-C₃Alkyl), O, S or Se,

◆A₂is N, CH or C (R)₅)，

G is selected from:

-C(O)OR_G3、-C(O)NR_G1R_G2、-C(O)R_G2、-NR_G1C(O)R_G2、-NR_G1C(O)NR_G1R_G2、-OC(O)NR_G1R_G2、-NR_G1C(O)OR_G3、-C(＝NOR_G1)NR_G1R_G2、-NR_G1C(＝NCN)NR_G1R_G2、-NR_G1S(O)₂NR_G1R_G2、-S(O)₂R_G3、-S(O)₂NR_G1R_G2、-NR_G1S(O)₂R_G2、-NR_G1C(＝NR_G2)NR_G1R_G2、-C(＝S)NR_G1R_G2、-C(＝NR_G1)NR_G1R_G2halogen, -NO₂and-CN, wherein:

-R_G1and R_G2Independently of one another at each occurrence, is selected from hydrogen, C optionally substituted by 1 to 3 halogen atoms₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₆Cycloalkyl, phenyl and- (CH)₂)_1-4-a phenyl group;

-R_G3selected from C optionally substituted by 1 to 3 halogen atoms₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₆Cycloalkyl, phenyl and- (CH)₂)_1-4-a phenyl group; or

R_G1And R_G2Together with the atoms to which they are simultaneously attached form C₃-C₈A heterocycloalkyl group; alternatively, G is selected from:

wherein R is_G4Selected from C optionally substituted by 1 to 3 halogen atoms₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C ₂-C₆Alkynyl and C₃-C₆A cycloalkyl group,

◆R₄represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group,

◆R₅represents a group selected from: c optionally substituted by 1 to 3 halogen atoms₁-C₆An alkyl group; c₂-C₆An alkenyl group; c₂-C₆An alkynyl group; a halogen or a group-CN,

◆R₆represents a group selected from:

hydrogen;

-C₂-C₆an alkenyl group;

-X₂-O-R₇；

-X₂-NSO₂-R₇；

-C＝C(R₉)-Y₁-O-R₇；

C₃-C₆a cycloalkyl group;

c optionally substituted by hydroxy₃-C₆A heterocycloalkyl group;

C₃-C₆cycloalkylene-Y₂-R₇；

C₃-C₆heterocycloalkylene-Y₂-R₇，

C optionally substituted by straight or branched chain₁-C₆Alkyl substituted heteroarylene-R₇，

◆R₇Represents a group selected from: straight or branched C₁-C₆An alkyl group; (C)₃-C₆) cycloalkylene-R₈(ii) a Or:

wherein Cy represents C₃-C₈A cycloalkyl group,

◆R₈represents a group selected from: hydrogen; straight or branched C₁-C₆Alkyl, -NR'_aR’_b；-NR’_a-CO-OR’_c；-NR’_a-CO-R’_c；-N⁺R’_aR’_bR’_c；-O-R’c；-NH-X’₂-N⁺R’_aR’_bR’_c；-O-X’₂-NR’_aR’_b、-X’₂-NR’_aR’_b、-NR’_c-X’₂-N₃And

◆R₉represents a group selected from: straight or branched C₁-C₆Alkyl, trifluoromethyl, hydroxy, halogen, C₁-C₆An alkoxy group,

◆R₁₀represents a group selected from: hydrogen, fluorine, chlorine, bromine, -CF₃And a methyl group,

◆R₁₁represents a group selected from: hydrogen, halogen, C₁-C₃alkylene-R₈、-O-C₁-C₃alkylene-R₈、-CO-NR_hR_iand-CH ═ CH-C₁-C₄alkylene-NR_hR_i、-CH＝CH-CHO、C₃-C₈cycloalkylene-CH₂-R₈、C₃-C₈heterocycloalkylene-CH₂-R₈，

◆R₁₂And R₁₃Independently of one another, represents a hydrogen atom or a methyl group,

◆R₁₄and R₁₅Independently of one another, represents hydrogen or methyl, or R₁₄And R ₁₅Form cyclohexyl with the carbon atom carrying them,. diamond-solid.R_hAnd R_iIndependently of one another, represents hydrogen or straight-chain or branched C₁-C₆An alkyl group, a carboxyl group,

◆X₁represents a straight or branched chain C₁-C₄Alkylene radical of anyOptionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C₁-C₆The substituent of the alkoxy group is replaced by the group,

◆X₂represents a straight or branched chain C₁-C₆Alkylene optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C₁-C₆The substituent of the alkoxy group is replaced by the group,

◆X’₂represents a straight or branched chain C₁-C₆An alkylene group or a substituted alkylene group,

◆R’_aand R'_bIndependently of one another, represent a group selected from: hydrogen; a heterocycloalkyl group; -SO₂Phenyl, wherein the phenyl radical may be linear or branched C₁-C₆Alkyl substitution; optionally substituted by one or two hydroxy groups or C₁-C₆Alkoxy-substituted straight or branched C₁-C₆An alkyl group; c₁-C₆alkylene-SO₂OH；C₁-C₆alkylene-SO₂O^-；C₁-C₆alkylene-COOH; c₁-C₆alkylene-PO (OH)₂；C₁-C₆alkylene-NR'_dR’_e；C₁-C₆alkylene-N⁺R’_dR’_eR’_f；C₁-C₆alkylene-O-C₁-C₆alkylene-OH; c₁-C₆Alkylene-phenyl in which the phenyl group may be substituted by hydroxy or C₁-C₆Alkoxy substitution;

the following groups:

or R'_aAnd R'_bForm a ring B with the nitrogen atom carrying them₃，

Or R'_a、R’_bAnd R'_cForm a bridge C with the nitrogen atom carrying them₃-C₈A heterocycloalkyl group, a heterocyclic alkyl group,

◆R’_c、R’_d、R’_e、R’_findependently of one another, represents hydrogen or straight-chain or branched C ₁-C₆An alkyl group, which is a radical of an alkyl group,

or R'_dAnd R'_eForm a ring B with the nitrogen atom carrying them₄，

Or R'_d、R’_eAnd R'_fForm a bridge C with the nitrogen atom carrying them₃-C₈A heterocycloalkyl group, a heterocyclic alkyl group,

◆Y₁represents a straight or branched chain C₁-C₄An alkylene group or a substituted alkylene group,

◆Y₂represents a bond, -O-CH₂-、-O-CO-、-O-SO₂-、-CH₂-、-CH₂-O、-CH₂-CO-、-CH₂-SO₂-、-C₂H₅-、-CO-、-CO-O-、-CO-CH₂-、-CO-NH-CH₂-、-SO₂-、-SO₂-CH₂-、-NH-CO-、-NH-SO₂-，

M is 0, 1 or 2,

p is 1, 2, 3 or 4,

◆B₁、B₂、B₃and B₄Independently of one another represent C₃-C₈Heterocycloalkyl, which group may be: (i) is a monocyclic or bicyclic group, wherein bicyclic group comprises a fused, bridged or spiro ring system, (ii) may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen in addition to the nitrogen atom, (iii) is substituted with one or two groups selected from: fluorine, bromine, chlorine, straight or branched C₁-C₆Alkyl, hydroxy, -NH₂An oxo group or a piperidyl group,

it should also be understood that:

- "aryl" means phenyl, naphthyl, biphenyl or indenyl,

- "heteroaryl" means any mono-or bicyclic group consisting of 5 to 10 ring members, having at least one aromatic moiety and containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen (including quaternary nitrogen),

- "cycloalkyl" means any mono-or bicyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, which may include fused, bridged or spiro ring systems,

- "heterocycloalkyl" means any mono-or bicyclic non-aromatic carbocyclic group consisting of 3 to 10 ring members containing 1 to 3 members selected from oxygen, sulfur, SO₂And nitrogen, it being understood that the bicyclic group may be fused or spiro,

heteroarylene, cycloalkylene, heterocycloalkylene refer to divalent heteroaryl, cycloalkyl and heterocycloalkyl groups, their enantiomers and diastereomers, and their addition salts with a pharmaceutically acceptable acid or base.

Among the pharmaceutically acceptable acids, mention may be made, without implying any limitation, of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphonic acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid, oxalic acid, methanesulfonic acid and camphoric acid.

Among the pharmaceutically acceptable bases, sodium hydroxide, potassium hydroxide, triethylamine and tert-butylamine may be mentioned, without implying any limitation.

Further exemplified embodiment (E) of the invention is described herein. It will be appreciated that the features specified in each embodiment may be combined with other specified features to provide further embodiments of the invention.

E2.A compound according to E1, which is a compound of formula (IA):

E3.A compound according to E1 or E2, wherein Z₁represents-NH-or-O-.

E4.A compound according to any one of E1 to E3, wherein R₃represents-X₁-NR_aR_bPreference is given to the group-C₂H₅-NH-CH₃。

E5.A compound according to E1 or E2, selected from:

E6.a compound according to E5, which is a compound of formula (IB):

E7.a compound according to E6, wherein Z₁Represents a bond and R₃Represents a hydrogen atom.

E8.A compound according to E1, which is a compound of formula (IC):

wherein A is₃Represents an oxygen or sulfur atom.

E9.A compound according to any one of E1 to E8, wherein R₁Represents a hydrogen atom, a methyl group or a cyclopropyl group, preferably a methyl group.

E10.A compound according to any one of E1 to E9, wherein Het is₁Represents:

E11.a compound according to any one of E1 to E10, wherein Het is₂Represents:

E12.a compound according to any one of E1 to E10, wherein Het is₂Represents:

E13.a compound according to E11, wherein R₆represents-X₂-O-R₇Wherein X is₂Is a propylene group.

E14.A compound according to E13, wherein R₇Represents the following groups:

E15.a compound according to E13, wherein R₇Represents the following groups:

E16.a compound according to E13, wherein R₇Represents the following groups:

E17.a compound according to any one of E14 to E16, wherein R₈Represents NR'_aR’_b。

E18.A compound according to any one of E14 to E16, wherein R₈Represents a group selected from: dimethylamino, diethylamino, diisopropylamino, diisobutylamino, methylamino, ethylamino, ethyl (methyl) amino, 4-methyl-piperazin-1-yl, pyrrolidin-1-yl, azetidin-1-yl, 1-piperidinyl, 4-morpholinyl, 4-difluoropiperidin-1-yl, 3-difluoropiperidin-1-yl, 3-hydroxy-1-piperidinyl, (1S,5R) -3-azabicyclo [3.1.0 ]Hex-3-yl, 4- (1-piperidinyl) -1-piperidinyl, 3-oxo-2, 8-diazaspiro [4.5 ]]Decan-8-yl, (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl, 2- (dimethylamino) ethylamino, 3-piperazin-1-yl, (3R, 5)S) -3, 5-dimethylpiperazin-1-yl, (but-3-yn-1-yl) amino, (but-3-yn-1-yl) (methyl) amino, (3-azidopropyl) (methyl) amino (3-aminopropyl) amino, (pent-4-yn-1-yl) amino, methyl (pent-4-yn-1-yl) amino, (prop-2-yn-1-yl) amino, (hex-5-yn-1-yl) amino, 3- [ (hex-5-yn-1-yl) (methyl) amino, (4-azidobutyl) (methyl) amino, di-or tri-amino, [2- (2-Hydroxyethoxy) ethyl group](methyl) amino and

E19.a compound according to any one of E14 to E16, wherein R₈Represents a group selected from: bis [ (3S) -3, 4-dihydroxybutyl)]Amino, [ (3S) -3, 4-dihydroxybutyl]Amino, [ (3R) -3, 4-dihydroxybutyl]Amino, acetyl (methyl) amino, 3-hydroxypropylamino.

E20.A compound according to E13, wherein R₇Represents:

wherein R is₁₁Selected from the group consisting of 3- (dimethylamino) propyl, 3- (methylamino) propyl, aminomethyl, 2- (dimethylamino) ethyl, 4- (dimethylamino) butyl, 2- (methylamino) ethyl, 4- (methylamino) butyl, 3- (azetidin-1-yl) propyl, 3- (4-methylpiperazin-1-yl) propyl, 3-pyrrolidin-1-ylpropyl, 3-morpholinopropyl, 3- (1-piperidinyl) propyl, 3- [ (1R,5S) -6, 6-difluoro-3-azabicyclo [3.1.0 ]Hex-3-yl and 3- (3-oxo-2, 8-diazaspiro [4.5 ]]Decan-8-yl) propyl.

E21.A compound according to E13, wherein R₇Represents a group selected from:

E22.a compound according to E12, wherein R₆Represents:

E23.a compound according to E22, wherein R₇Represents a group selected from:

wherein R is₈represents-O-X'₂-NR’_aR’_bor-X'₂-NR’_aR’_b。

E24.A compound according to E22, wherein R₇Represents a group selected from:

wherein R is₈Represents a group selected from: hydrogen, 2- (methylamino) ethoxy, 2- (dimethylamino) ethoxy, 2- [ (2-sulfoethyl) amino]Ethoxy, 2- [ methyl (2-sulfoethyl) amino]Ethoxy, 4-methylpiperazin-1-yl and

E25.a compound according to E22, wherein R₇Represents a group selected from:

wherein R is₈Represents a group selected from: 2-pyrrolidin-1-ylethoxy, 2- (4-methylpiperazin-1-yl) ethoxy, 2- [, [ solution of ] A(3R) -3, 4-Dihydroxybutyl]-methyl-amino]Ethoxy, 2- (4-hydroxybutylamino) ethoxy, 2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] ethyl]Amino group]Ethoxy, 2- [ bis (2-hydroxyethyl) amino]Ethoxy, 2- [ [ 2-hydroxy-1- (hydroxymethyl) ethyl]Amino group]Ethoxy, 2- [2- (2-hydroxyethoxy) ethylamino]Ethoxy, 2- [ bis (3-hydroxypropyl) amino]Ethoxy, 2- (3-hydroxypropylamino) ethoxy, 2- [ bis (4-hydroxybutyl) amino ]Ethoxy, 2-morpholinoethoxy, 2- (1-piperidinyl) ethoxy, 2-piperazin-1-ylethoxy, 2- (azepan-1-yl) ethoxy, 2- (4-isopropylpiperazin-1-yl) ethoxy, 2- [ (4-hydroxyphenyl) methylamino]Ethoxy, 2- [ 2-hydroxyethyl (methyl) amino]Ethoxy, 2- [ 3-methoxypropyl (methyl) amino]Ethoxy, 2- [ 4-hydroxybutyl (methyl) amino]Ethoxy, 3-pyrrolidin-1-ylpropyl, 3- (dimethylamino) propyl, 3- (4-methylpiperazin-1-yl) propyl, 3-morpholinopropyl, 3- (3-hydroxypropylamino) propyl, 3- (4-hydroxybutylamino) propyl, 3- [ [ (3S) -3, 4-dihydroxybutylamino ] propyl]Amino group]Propyl, 3-hydroxy-2- (hydroxymethyl) propyl]Amino group]Propyl, 3- [ 4-hydroxybutyl (methyl) amino]Propyl, 3- [ 3-hydroxypropyl (methyl) amino]Propyl, 3- [3- [ bis (3-hydroxypropyl) amino]Propyl, 3-piperazin-1-ylpropyl.

E26.A compound according to any one of E1, E2 and E6, wherein R₃represents-X₁-PO(OH)₂、-X₁-SO₂(OH)、-X₁-NR_aR_b；-X₁-N⁺R_aR_bR_cWherein R is_aOr R_bOr R is_aAnd R_bBoth of them represent a group selected from C₁-C₆alkylene-SO₂OH、C₁-C₆alkylene-SO₂O^-And C₁-C₆alkylene-PO (OH)₂A group of (1).

E27.A compound according to any one of E1, E2 and E6, wherein R₈represents-NR'_aR’_b；-N⁺R’_aR’_bR’_c；-NH-X’₂-N⁺R’_aR’_bR’_cWherein R'_aAnd R'_bOr both represent C ₁-C₆alkylene-SO₂OH and C₁-C₆alkylene-PO (OH)₂A group of (2).

E28.A compound according to E1, selected from:

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [3- (dimethylamino) prop-1-ynyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ] pyridazin-8-yl } -5- (3- { 2-fluoro-4- [3- (methylamino) prop-1-yn-1-yl ] phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid,

-2- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ] pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) propyl ] -2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) but-1-ynyl ] phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylprop-1-ynyl) phenoxy ] propyl ] thiazole-4-carboxylic acid,

-5- (3- {4- [3- (azetidin-1-yl) prop-1-yn-1-yl ] -2-fluorophenoxy } propyl) -2- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ] pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) prop-1-ynyl ] phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ] pyridazin-8-yl } -5- (3- {4- [3- (4, 4-difluoropiperidin-1-yl) prop-1-yn-1-yl ] -2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid,

-2- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ] pyridazin-8-yl } -5- (3- {4- [3- (3, 3-difluoropiperidin-1-yl) prop-1-yn-1-yl ] -2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- [3- (3-oxo-2, 8-diazaspiro [4.5] decan-8-yl) prop-1-ynyl ] phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [3- [ (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0] hex-3-yl ] prop-1-ynyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- (3-piperazin-1-ylprop-1-ynyl) phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [3- [ (3R,5S) -3, 5-dimethylpiperazin-1-yl ] prop-1-ynyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [3- (diethylamino) prop-1-ynyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [3- (diisopropylamino) prop-1-ynyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [3- [2- (dimethylamino) ethylamino ] prop-1-ynyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-6- [2- (methylamino) ethoxy ] -5H,6H,7H, 8H-pyrido [2,3-c ] pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl ] -2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [1- [ (dimethylamino) methyl ] -3-bicyclo [1.1.1] pentyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- [ 3-methyl-3- (methylamino) but-1-ynyl ] phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- [3- (prop-2-ynylamino) prop-1-ynyl ] phenoxy ] propyl ] thiazole-4-carboxylic acid,

-6- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-6, 7-dihydropyrido [2,3-c ] pyridazin-8 (5H) -yl } -3- [1- ({3- [2- (dimethylamino) ethoxy ] -5, 7-dimethyladamantan-1-yl } methyl) -5-methyl-1H-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-6, 7-dihydropyrido [2,3-c ] pyridazin-8 (5H) -yl } -3- [1- ({3, 5-dimethyl-7- [2- (methylamino) ethoxy ] adamantan-1-yl } methyl) -5-methyl-1H-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-2- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-6, 7-dihydropyrido [2,3-c ] pyridazin-8 (5H) -yl } -5- (3- {4- [3- (ethylamino) -3-methylbut-1-yn-1-yl ] -2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid,

-3- {1- [ (adamantan-1-yl) methyl ] -5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ] pyridazin-8-yl } pyridine-2-carboxylic acid,

Enantiomers and diastereomers thereof, and addition salts thereof with a pharmaceutically acceptable acid or base.

E29.A compound according to E1, selected from:

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3, 5-dimethyl-7- [2- (4-methylpiperazin-1-yl) ethoxy ] -1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- (3-hydroxypropylamino) ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- (4-hydroxybutylamino) ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-6, 7-dihydropyrido [2,3-c ] pyridazin-8 (5H) -yl } -3- (1- { [3- (2- { [ (3S) -3, 4-dihydroxybutyl ] amino } ethoxy) -5, 7-dimethyladamantan-1-yl ] methyl } -5-methyl-1H-pyrazol-4-yl) pyridine-2-carboxylic acid,

-6- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-6, 7-dihydropyrido [2,3-c ] pyridazin-8 (5H) -yl } -3- (1- { [3- (2- { [ (3R) -3, 4-dihydroxybutyl ] amino } ethoxy) -5, 7-dimethyladamantan-1-yl ] methyl } -5-methyl-1H-pyrazol-4-yl) pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ 2-hydroxyethyl (methyl) amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ 4-hydroxybutyl (methyl) amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ [ (3R) -3, 4-dihydroxybutyl ] -methyl-amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3, 5-dimethyl-7- (2-piperazin-1-ylethoxy) -1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3, 5-dimethyl-7- [2- (1-piperidinyl) ethoxy ] -1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-3- [1- [ [3- [2- (azepan-1-yl) ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] -6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- (4-isopropylpiperazin-1-yl) ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3, 5-dimethyl-7- (2-morpholinoethoxy) -1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ 3-methoxypropyl (methyl) amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [2- (2-hydroxyethoxy) ethylamino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ [ 2-hydroxy-1- (hydroxymethyl) ethyl ] amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ bis (2-hydroxyethyl) amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ bis (3-hydroxypropyl) amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ bis (4-hydroxybutyl) amino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-6- {3- [ (1, 3-benzothiazol-2-yl) amino ] -4-methyl-6, 7-dihydropyrido [2,3-c ] pyridazin-8 (5H) -yl } -3- {1- [ (3, 5-dimethyl-7- {2- [ (2-sulfoethyl) amino ] ethoxy } adamantan-1-yl) methyl ] -5-methyl-1H-pyrazol-4-yl } pyridine-2-carboxylic acid,

-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -3- [1- [ [3- [2- [ (4-hydroxyphenyl) methylamino ] ethoxy ] -5, 7-dimethyl-1-adamantyl ] methyl ] -5-methyl-pyrazol-4-yl ] pyridine-2-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [4- [3- [ [ (3S) -3, 4-dihydroxybutyl ] amino ] prop-1-ynyl ] -2-fluoro-phenoxy ] propyl ] thiazole-4-carboxylic acid,

-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] pyridazin-8-yl ] -5- [3- [ 2-fluoro-4- [3- (3-hydroxypropylamino) prop-1-ynyl ] phenoxy ] propyl ] thiazole-4-carboxylic acid,

E30.A process for the preparation of a compound of formula (I) according to E6, characterized in that a compound of formula (II) is used as starting material:

introducing a leaving group (preferably iodinated) to a compound of formula (II) to produce a compound of formula (III):

wherein L.G represents a leaving group (preferably a halogen atom, more preferably iodine),

further subjecting the compound of formula (III) to a coupling reaction with a compound of formula (IV) in water or an organic medium, preferably acetone, in the presence of a base, preferably cesium carbonate:

wherein G1 represents C ₁-C₆Alkyl or (4-methoxyphenyl) methyl andrepresents a protecting group (preferably tert-butoxycarbonyl) to produce a compound of the formula (V)Material (b):

deprotecting the amino group of the compound of formula (V) (preferably using 1,1,1,3,3, 3-hexafluoroisopropanol) to produce a compound of formula (VI):

the compound of formula (VI) is reacted in aqueous or organic medium in a palladium phosphine complex, preferably Pd (AtaPhos)₂Cl₂) Alkali (preferably Cs)₂CO₃) And a Suzuki coupling reaction in the presence of a compound of formula (VII):

wherein R is₇As defined in formula (I),

to produce a compound of formula (VIII):

the compound of formula (VIII) is further reacted in an aqueous or organic medium in a palladium phosphine complex, preferably Pd (AtaPhos)₂Cl₂) And at least one base (preferably Cs)₂CO₃And DIPEA) to produce a compound of formula (IX):

the compound of formula (IX) is reacted in an aqueous or organic medium over a palladium catalyst, preferably Pd₂(dba)₃) Buchwald reaction in the presence of a base (preferably DIPEA), a phosphine (preferably Xantphos) and a compound of formula (X):

wherein R is₄And m is as defined for formula (I),

to produce a compound of formula (XI):

hydrolysis of the ester function of the compound of formula (XI) (preferably using LiOH H)₂O or TFA) to produce a compound of formula (I),

the compounds of formula (I) can be purified according to conventional isolation techniques, can be converted into addition salts thereof with pharmaceutically acceptable acids or bases, and optionally isolated as isomers thereof according to conventional isolation techniques,

It will be appreciated that at any time deemed appropriate during the above-described methods, the hydroxyl, amino, carboxyl and phosphono groups of the reagents or intermediates of the synthesis may be protected and then deprotected as required for the synthesis.

E31.The process according to E30, wherein the radical R₇Selected from:

wherein R is₈、R₁₂And R₁₃As defined in formula (I).

E32.A process for the preparation of a compound of formula (I) according to E6, characterized in that a compound of formula (II) is used as starting material:

a compound of formula (II) is subjected to a Mitsunobu reaction in toluene in the presence of triphenylphosphine, a suitable coupling agent, preferably di-tert-butyl azodicarboxylate, and a compound of formula (XII-a) or (XII-b):

wherein A is₁、A₂And R₆As defined in formula (I), G1 represents C₁-C₆Alkyl or (4-methoxyphenyl) methyl andrepresents a protecting group (preferably tert-butoxycarbonyl),

to produce a compound of formula (XIII-a) or (XIII-b):

deprotecting the amino group of the compound (XIII-a) or (XIII-b) to produce a compound of formula (XIV-a) or (XIV-b):

(i) subjecting a compound of formula (XIV-a) to a base, preferably Cs₂CO₃) Further performing an intramolecular coupling reaction in an aqueous or organic medium in the presence of a catalyst to produce a compound of formula (XV-a),

or

(ii) The compound of formula (XIV-b) is further reacted in an aqueous or organic medium in a palladium phosphine complex, preferably Pd (AtaPhos) ₂Cl₂) And at least one base (preferably Cs)₂CO₃And DIPEA) to produce a compound of formula (XV-b):

the compound of formula (XV-a) or (XV-b) is catalyzed by palladium in an aqueous or organic mediumAgent (preferably Pd)₂(dba)₃) Buchwald reaction in the presence of a base (preferably DIPEA), a phosphine (preferably Xantphos) and a compound of formula (X):

to produce a compound of formula (XVI-a) or (XVI-b):

hydrolysis of the ester function of the compound of formula (XVI-a) or (XVI-b) (preferably using LiOH XH)₂O or TFA) to produce a compound of formula (I),

E33.A synthetic intermediate according to E30 or E31 selected from:

wherein R is₇As defined in formula (I) and G1 represents C₁-C₆Alkyl, preferably methyl, or (4-methoxyphenyl) methyl.

E34.A synthetic intermediate according to E32, selected from:

wherein R is₆As defined in formula (I) and G1 represents C ₁-C₆Alkyl, preferably methyl, or (4-methoxyphenyl) methyl.

E35.A compound according to E1, wherein R₄Represents a hydrogen, fluorine, chlorine or bromine atom, a methyl or methoxy group.

E36.A compound according to E1, wherein R₈Represents a group selected from: hydrogen; straight or branched C₁-C₆Alkyl, -NR'_aR’_b；-NR’_a-CO-OR’_c；-N⁺R’_aR’_bR’_c；-O-R’c；-NH-X’₂-N⁺R’_aR’_bR’_c；-O-X’₂-NR’_aR’_b、-NR’_c-X’₂-N₃And

E37.a compound according to E1, wherein R'_aAnd R'_bIndependently of one another, represent a group selected from: hydrogen; a heterocycloalkyl group; -SO₂Phenyl, wherein the phenyl radical may be linear or branched C₁-C₆Alkyl substitution; straight or branched chain C optionally substituted by one or two hydroxy groups₁-C₆An alkyl group; c₁-C₆alkylene-SO₂OH；C₁-C₆alkylene-SO₂O^-；C₁-C₆alkylene-COOH; c₁-C₆alkylene-PO (OH)₂；C₁-C₆alkylene-NR'_dR’_e；C₁-C₆alkylene-N⁺R’_dR’_eR’_f；C₁-C₆alkylene-O-C₁-C₆alkylene-OH; c₁-C₆Alkylene-phenyl, in which the phenyl radical may be substituted by C₁-C₆Alkoxy substitution;

the following groups:

or R'_aAnd R'_bForm a ring B with the nitrogen atom carrying them₃，

Or R'_a、R’_bAnd R'_cForm a bridge C with the nitrogen atom carrying them₃-C₈A heterocycloalkyl group.

E38.A compound according to any one of E1 to E27, wherein m ═ 1.

Pharmacological studies with the compounds of the invention show that they possess pro-apoptotic properties. The ability to reactivate apoptotic processes in cells is of great therapeutic interest in the treatment of cancer and immune and autoimmune diseases. In particular, the compounds according to the invention will be useful in the treatment of chemo-or radiation-resistant cancers.

In another embodiment, the compounds of the invention are useful in the treatment of diseases or conditions characterized by excessive or deregulated platelet activity, particularly pre-thrombotic conditions.

As used herein, the term "treatment" of any disease or condition refers in one embodiment to ameliorating the disease or condition (i.e., slowing or arresting or reducing the development of the disease or at least one clinical symptom thereof). In another embodiment, "treating" or "treatment" refers to reducing or improving at least one physical parameter, including those that may not be discernible by the patient. In yet another embodiment, "treating" or "treatment" refers to modulating the disease or disorder, either physically (e.g., stabilization of a discernible symptom), physiologically (e.g., stabilization of a physical parameter), or both.

Among the cancer treatments envisaged, mention may be made, without implying any limitation, of the treatment of haematological malignancies and solid tumours. Hematological malignancies include myelomas, particularly multiple myeloma, lymphomas, particularly non-hodgkin's lymphoma (NHL), more particularly diffuse large B-cell lymphoma (DLBCL), and leukemias, particularly Chronic Lymphocytic Leukemia (CLL), acute T-lymphocytic leukemia (T-ALL), acute B-lymphocytic leukemia (B-ALL), and Acute Myeloid Leukemia (AML). Solid tumors include bladder, brain, breast, uterus, duct and liver cancers, colorectal, kidney, melanoma, ovarian, prostate, pancreatic and lung cancers, especially non-small cell and small cell lung cancers.

In particular, T-ALL is caused by leukemic transformation of thymocyte precursors and their arrest at specific stages of differentiation. Despite recent insights into the molecular and cellular mechanisms that lead to the pathogenesis and progression of T-ALL, these knowledge have not yet translated into effective targeted therapies. Current clinical treatments include chemotherapy with or without hematopoietic stem cell transplantation, with survival rates in adult and pediatric cases remaining around 50% and 70%, respectively. In both pediatric and adult cases, relapse shows very poor prognosis, which enhances the need to discover new treatment options (Pasaro et al, Immunol Rev.2016 May; 271 (1): 156-72). Bcl-2/Bcl-xL dual inhibitors, such as ABT-263 and ABT-737, have been shown to have promising activity in T-ALL patient-derived xenograft models (Van Delft et al, Cancer Cell 2006; 10: 389-99; Suryani et al, Clin Cancer Res.2014, 20: 4520-31). Other studies reported different needs for mature and very immature (ETP subgroup) T-ALL survival for Bcl-xL or Bcl-2 (Chonghaile et al, Cancer Discov.2014; 4: 1074-87). The previously described selective Bcl-xL inhibitor A-1331852 has also been shown to have in vitro and in vivo activity in the mature T-ALL cell line xenograft model Molt-4 (Leverson et al, Sci Transl Med.2015 Mar 18; 7(279):279ra 40). In a specific embodiment, tumor growth inhibition is also observed in MOLT-4 xenograft models following treatment with a Bcl-xL inhibitor of the invention. These data support the use of the compounds of the invention in the treatment of T-ALL.

Among the treatments envisaged for autoimmune diseases, mention may be made, without implying any limitation, of the treatment of Rheumatoid Arthritis (RA) and Systemic Lupus Erythematosus (SLE).

The invention also relates to pharmaceutical compositions comprising at least one compound of formula (I) as active ingredient, together with one or more pharmaceutically acceptable excipients. In particular, the use of these pharmaceutical compositions as pro-apoptotic and/or anti-proliferative agents is of interest, particularly in the treatment of cancer and autoimmune and immune system diseases.

Suitable excipients according to the invention include diluents, lubricants, binders, disintegrants, stabilizers, preservatives, absorbents, colorants, sweeteners and flavoring agents.

As non-limiting examples, mention may be made of:

as diluents: lactose, glucose, sucrose, mannitol, sorbitol, cellulose, glycerol,

as lubricant: silicon dioxide, talc, stearic acid and its magnesium and calcium salts, polyethylene glycol,

as binder: magnesium aluminum silicate, starch, gelatin, tragacanth, methyl cellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone,

as disintegrant: agar, alginic acid and sodium salt thereof, and effervescent mixture.

Among the pharmaceutical compositions according to the invention, mention may be made more particularly of those suitable for oral, parenteral, nasal, transdermal or transdermal, rectal, lingual, ocular or respiratory administration, in particular tablets, dragees, sublingual tablets, capsules, glossettes, capsules, lozenges, injectable or drinkable preparations, aerosols, eye or nasal drops, suppositories, creams, ointments, dermal gels.

Actual dosage levels of the active ingredient in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration without being toxic to the patient. The selected dosage level will depend upon a variety of factors including the activity of the particular compound of the invention employed, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound employed, the rate and extent of absorption, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

Suitable daily dosages of the compounds of the invention will depend on the factors mentioned above and are in the range of from 0.01mg to 2.5g per day, administered in one or more doses.

In another aspect, the invention also relates to the combination of a compound of formula (I) with an anticancer agent selected from genotoxic agents, mitotic toxic agents, antimetabolites, proteasome inhibitors, kinase inhibitors and antibodies, and also to pharmaceutical compositions comprising such combinations and their use in the manufacture of medicaments for the treatment of cancer.

In another aspect, the compounds of the invention may be used in combination with radiation therapy for the treatment of cancer.

Alternatively, the compounds of the invention may be linked to monoclonal antibodies. Antibody Drug Conjugates (ADCs) represent a new class of therapeutic agents that are formed by chemically linking a cytotoxic drug to a monoclonal antibody through a linker. The monoclonal antibody of the ADC selectively binds to a target antigen of a cell (e.g., a cancer cell) and releases the drug into the cell. ADCs have therapeutic potential because they combine the specificity of antibodies and the cytotoxic potential of drugs. Nevertheless, development of ADCs as therapeutic agents has to date had only limited success due to factors such as adverse toxicity profiles, low efficacy and poor pharmacological parameters. Thus, there remains a need for new ADCs that overcome these problems and can selectively deliver Bcl-xL to target cancer cells.

In another aspect, the compounds of the invention may be linked to fragments of a monoclonal antibody or to a scaffold protein, which may or may not be associated with a monoclonal antibody. Antibody fragments must be understood as being fragments of Fv, scFv, Fab, F (ab ')2, F (ab'), scFv-Fc type or diabodies, which generally have the same binding specificity as the antibody from which they originate. According to the present invention, the antibody fragment of the present invention can be obtained by a method such as enzymatic digestion, e.g., pepsin or papain digestion, and/or cleavage of disulfide bonds by chemical reduction, starting from the antibody. In another mode, the antibody fragments encompassed by the present invention may be obtained by genetic recombination techniques also well known to those skilled in the art, or by peptide synthesis, such as those provided by automated peptide synthesizers such as Applied Biosystems, and the like.

A scaffold protein that may or may not be associated with a monoclonal antibody is understood to mean a protein that contains or does not contain immunoglobulin folds and that produces similar binding capacity as a monoclonal antibody. The person skilled in the art knows how to select a protein scaffold. More particularly, it is known that the selected scaffold should exhibit several characteristics (Skerra, j.mol.recogn.2000, 13, 167-: phylogenetically well conserved, robust structure with well-known three-dimensional molecular organization (e.g. crystallography or NMR), small size, no or only low degree of post-translational modification, ease of production, expression and purification. Such a protein scaffold may be, but is not limited to, a protein Z derivative selected from fibronectin, preferably the tenth fibronectin type III domain (FNfn10), lipocalins, anticalin (Skerra, j. biotechnol.2001,74,257-75), domain B of staphylococcal protein a, thioredoxin a or any protein with a repeat domain such as "ankyrin repeat" (Kohl et al PNAS 2003,100,1700-1705), "armadillo repeat", "leucine rich repeat" or "tetratriaconteptide (tetratricopeptide) repeat". Also mentioned are scaffold derivatives derived from toxins such as scorpion, insect, plant or mollusc toxins, or protein inhibitors of neuronal nitric oxide synthase (PIN).

The following examples illustrate the invention but do not limit it in any way. All intermediates used in the preparation of the examples are commercially available or can be obtained by one skilled in the art using conventional chemical reactions described in the literature.

General procedure

All reagents obtained from commercial sources were used without further purification. The anhydrous solvent was obtained from a commercial source and used without further drying.

Column chromatography

Use ofRf normal phase silica gel flash column (35-70 μm,)、RediSep Rfnormal phase silica gel high performance column (20-40 μm,)、rf reversed phase C18 column (40-63m,) Or RediSep RfThe reversed phase C18 high performance column (20-40m,) In ISCORf 200 orRf+Lumen^TMThe automated flash column chromatography was performed.

Thin Layer Chromatography (TLC)

Merck Type 60F for thin layer chromatography₂₅₄Silica gel coated 5X 10cm plates.

Microwave reaction

By CEMSP or single-wave microwave with Anton PaarThe wave reactor is heated by microwave.

Nuclear Magnetic Resonance (NMR)

DMSO-d on a Bruker Avance III 500MHz spectrometer, a Bruker Avance III 400MHz spectrometer or a Bruker DPX-400 spectrometer₆Or CDCl₃1H-NMR measurement was carried out as a solvent. 1H NMR data is in delta form, given in parts per million (ppm), using the residual peak of the solvent (DMSO-d)₆2.50ppm, CDCl ₃7.26ppm) as an internal standard. The split pattern is specified as: s (singlet), d (doublet), t (triplet), q (quartet), quant (quintet), sept (heptaplex), m (multiplet), br s (broad singlet), dd (doublet of doublets), td (triplet), dt (doublet of triplets), ddd (doublet of doublets).

Analytical LC-MS

Certain compounds of the invention were characterized by high performance liquid chromatography-mass spectrometry (HPLC-MS) on Agilent HP1200 operating with Agilent 6140 quadrupole LC/MS in positive or negative ion electrospray ionization mode. The molecular weight scan ranges from 100 to 1350. Parallel UV detection was performed at 210nm and 254 nm. The sample is treated with 1mM ACN solution or THF/H₂O (1:1) solution was provided as a 5 μ L loop injection. LCMS analysis was performed on two instruments, one operating with a basic eluent and the other operating with an acidic eluent.

Basic LCMS: Gemini-NX,3 μm, C18,50mM x 3.00mM i.d. column, flow rate 1 mL/min at 23 ℃, using 5mM ammonium bicarbonate (solvent A) and acetonitrile (solvent B), gradient starting from 100% solvent A and ending with 100% solvent B for different/specific durations.

Acidic LCMS: kinetex XB-C18-100A, 2.6m, 50mm x 2.1mm column, 40 ℃, flow rate 1 mL/min, gradient starting with 100% solvent a and ending with 100% solvent B over different/specified durations using 0.02% v/v formic acid in water (solvent a) and 0.02% v/v formic acid in acetonitrile (solvent B).

Certain other compounds of the invention were characterized by HPLC-MS using the following specific nomenclature. For all of these methods, UV detection was performed at 230, 254 and 270nm by diode array detectors. The sample injection volume was 1L, and the gradient elution was performed by defining the flow rate and the following percentage of the mixture of mobile phases, using HPLC grade solvents:

solvent A: 10mM aqueous ammonium formate + 0.04% (v/v) formic acid

Solvent B: acetonitrile + 5.3% (v/v) solvent A + 0.04% (v/v) formic acid.

The Retention Times (RT) of these named methods are reported in minutes. Ionization is recorded in positive, negative, or positive-negative switching mode. The following are specific details of each method.

LCMS-V-B method

Agilent 1200SL series instruments connected to a single quadrupole of Agilent MSD 6140 with ESI-APCI multimode source (methods LCMS-V-B1 and LCMS-V-B2) or Agilent 1290 Infinity II series instruments connected to Agilent TOF 6230 with ESI-jet source (method LCMS-V-B1); column: thermo Accucore 2.6m, C18, 50 mm. times.2.1 mm, 55 ℃. Details of gradient for methods LCMS-V-B1 and LCMS-V-B2:

LCMS-V-C process

Using an Agilent 1200SL series instrument connected to an Agilent MSD 6140 single quadrupole with ESI-APCI multimode source; column: agilent Zorbax Eclipse plus 3.5m, C18(2), 30mm x 2.1mm, 35 ℃. Gradient details of method LCMS-V-C:

Preparative HPLC

Certain compounds of the present invention were purified by High Performance Liquid Chromatography (HPLC) on an Armen Spot or Teledyne EZ system using Gemini-10μM C18,250mM × 50mM i.d. column, UV diode array detection (210- & 400nm), using 25mM NH₄HO₃Aqueous solution and MeCN or 0.1% TFA aqueous solution and MeCN as eluents.

Certain other compounds of the invention were purified by HPLC using the following specific nomenclature:

HPLC-V-A method

The method was performed on a Waters FractionLynx MS automated purification system using a product from Phenomenex5 μm C18(2),100mm × 20mm i.d. column, at 20cm³ min^-1Flow rate operation, UV diode array detection (210-400nm) and mass directed collection. The mass spectrometer was a Waters Micromass ZQ2000 mass spectrometer operating in either positive or negative ion electrospray ionization mode with a molecular weight scan range of 150-.

Method HPLC-V-a1(pH 4):

solvent A: 10mM aqueous ammonium acetate + 0.08% (v/v) formic acid; solvent B: acetonitrile + 5% (v/v) solvent A + 0.08% (v/v) formic acid

Method HPLC-V-a2(pH 9):

solvent A: 10mM ammonium acetate in water + 0.08% (v/v) concentrated ammonia; solvent B: acetonitrile + 5% (v/v) solvent A + 0.08% (v/v) concentrated ammonia water

HPLC-V-B method

Performed on an AccQPrep HP125(Teledyne ISCO) system using the reagent from PhenomenexNX 5. mu. m C18(2),150mm X21.2 mm i.d. column, at 20cm³min^-1Run with UV (214 and 254nm) and ELS detection.

Method HPLC-V-B1(pH 4):

solvent A: water + 0.08% (v/v) formic acid; solvent B: acetonitrile + 0.08% (v/v) formic acid.

Method HPLC-V-B2(pH 9):

solvent A: water + 0.08% (v/v) concentrated ammonia; solvent B: acetonitrile + 0.08% (v/v) concentrated ammonia.

Method HPLC-V-B3 (neutral):

solvent A: water; solvent B: and (3) acetonitrile.

Analytical GC-MS

The combination of gas chromatography and low resolution mass spectrometry (GC-MS) was performed on an Agilent 6850 gas chromatography and Agilent 5975C mass spectrometer using a 15 m.times.0.25 mm column, a 0.25 μm HP-5MS coating, with helium as the carrier gas. An ion source: EI +, 70eV, 230 ℃, quadrupole: 150 ℃, interface: at 300 ℃.

High resolution MS

High resolution mass spectra were obtained on an Agilent 6230 time-of-flight mass spectrometer equipped with a jet electrospray ion source in positive ion mode. Using an Agilent 1290 Infinity HPLC system, 0.5. mu.l of injection was introduced into the mass spectrometer at a flow rate of 1.5ml/min (5mM ammonium formate in water and acetonitrile gradient program). Jet parameters: drying gas (N2) flow and temperature: 8.0L/min and 325 ℃ respectively; nebulizer gas (N2) pressure: 30 psi; capillary voltage: 3000V; sheath airflow and temperature: 325 ℃ and 10.0L/min; TOFMS parameters: fragmentation voltage: 100V; potential of the skimmer: 60V; OCT 1 RF Vpp: 750V. Full scan mass spectra were acquired at an acquisition rate of 995.6 ms/spectrum in the m/z range 105-1700 and processed by the Agilent MassHunter B.04.00 software.

Chemical nomenclature

The IUPAC preferred name is the use of the Chemaxon's "Structure naming" (s2n) function in MarvinSketch or Jchem for Excel (Jchem version 16.6.13-18.22.3) or the use of the function consisting ofThe chemical naming function provided by Draw 4.2.

Abbreviations

Ahx 6-hexanoic acid monomer

AgOTf silver triflate

^tBuOH tert-butyl alcohol

cc. rich

CyOH cyclohexanol

dba (1E,4E) -1, 5-diphenylpenta-1, 4-dien-3-one, dibenzylideneacetone

DCM dichloromethane

DIPA N-isopropylpropan-2-amine, diisopropylamine

DIPEA N-ethyl-N-isopropyl-propan-2-amine, diisopropylethylamine

DMAP 4-dimethylaminopyridine

ee. enantiomeric filtration

eq. equivalent

EtOAc ethyl acetate

HF x Pyr pyridine hydrogen fluoride

hs intelligent

LDA lithium diisopropylamide

MeCN acetonitrile

MeOH methanol

NMP N-methyl-2-pyrrolidone

Pd(AtaPhos)₂Cl₂Bis (di-tert-butyl (4-dimethylaminophenyl) phosphine) palladium (II) dichloride

rt Room temperature

RT Retention time (minutes)

on overnight

TBAF tetrabutylammonium fluoride

TBAOH tetrabutylammonium hydroxide

TBDPS-Cl tert-butyl-chloro-diphenyl-silane

TBSCl tert-butyl-chloro-dimethyl-silane

TEA N, N-diethylethanamine

TFA 2,2, 2-trifluoroacetic acid

pTSA 4-methylbenzenesulfonic acid

THF tetrahydrofuran

TMP-MgCl 2,2,6, 6-tetramethyl piperidyl magnesium chloride lithium chloride complex solution

DIAD diisopropyl azodicarboxylate

Xantphos 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene

Brettphos 2- (dicyclohexylphosphine) -3, 6-dimethoxy-2 ', 4', 6 '-triisopropyl-1, 1' -biphenyl

Josiphos (2R) -1- [ (1R) -1- (dicyclohexylphosphine) ethyl ] -2- (diphenylphosphino) ferrocene

Josiphos Pd G3 { (R) -1- [ (Sp) -2- (dicyclohexylphosphine) ferrocenyl ] ethyldi-tert-butylphosphine } [2- (2 '-amino-1, 1' -biphenyl) ] palladium methanesulfonate (II)

Xantphos Pd G3 [ (4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene) -2- (2 '-amino-1, 1' -biphenyl) ] palladium methanesulfonate (II)

BINAP 2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl

rac-BINAP Pd G3 [ (2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl) -2- (2 '-amino-1, 1' -biphenyl) ] palladium (II) methanesulfonate

Pd(dppf)Cl₂.CH₂Cl₂[1, 1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (II)

Pd₂(dba)₃Tris (dibenzylideneacetone) dipalladium (0)

General method of naming

The following are representative experimental procedures, referred to by name in the preparations that follow.

Sonogashira general method

1 equivalent of aryl halide, 2 equivalents of acetylene, 0.05 equivalent of Pd (PPh)₃)₂Cl₂A mixture of 0.05 equivalents of CuI and DIPA (1mL/mmol) in THF (5mL/mmol) was maintained at 60 ℃. After reaching the appropriate conversion, the volatiles were removed under reduced pressure using heptaneThe crude intermediate was purified by flash chromatography using alkane/EtOAc as eluent.

General procedure for deprotection with HFIP

The substrate was stored in a pressure vial in HFIP (10mL/mmol) at 100 ℃ and 120 ℃. After the appropriate conversion was reached, the volatiles were removed under reduced pressure and the crude intermediate was purified by flash chromatography using heptane/EtOAc as eluent.

General procedure for deprotection and hydrolysis

A mixture of 1 equivalent of substrate and 100 equivalents of HFxPyr in MeCN (15mL/mmol) was stirred at 60 ℃. After the appropriate conversion was reached, the volatiles were removed under reduced pressure, the residue was suspended in a 1:1 mixture of THF-water (30mL/mmol), and 150 equivalents of LiOH x H were added ₂O, the mixture was stirred at room temperature. After reaching the appropriate conversion, the volatiles were removed under reduced pressure; the crude product was purified by flash chromatography using DCM and MeOH (containing 1.2% NH₃) As an eluent.

General alkylation process

1 equivalent of phenol/carbamate, 1 to 2 equivalents of alkyl iodide/bromine and 2 to 3 equivalents of Cs₂CO₃The mixture in acetone (5mL/mmol) was stirred at room temperature (for phenol) or at 55 deg.C (for carbamate). After the appropriate conversion was reached, the volatiles were removed under reduced pressure and the crude intermediate was purified by flash chromatography using heptane/EtOAc as eluent.

General procedure for alkylation with tosylates

In an oven-dried vial was equipped with a PTFE-coated magnetic stir bar, 1 equivalent of tosylate was added and 5 equivalents of the appropriate amine were suspended in MeCN (5 mL/mmol). The reaction mixture was then warmed to 50 ℃ and stirred at this temperature until no further conversion was observed. The reaction mixture was diluted with DCM and then poured onto a silica gel column pretreated with DCM. Then purified by flash chromatography using DCM and MeOH (1.2% NH₃) As an eluent.

MonosilaneGeneral process for alkylation of radical-protected phenols

A mixture of 1 equivalent of the silyl-protected phenol, 1 equivalent of the alkyl iodide, and 1.15 equivalents of TBAF (1M in THF) in THF (2mL/mmol) was stirred at room temperature. After the appropriate conversion was reached, the volatiles were removed under reduced pressure and the crude intermediate was purified by flash chromatography using heptane/EtOAc as eluent.

Buchwald general methodI

1 equivalent of chlorinated substrate, 2 equivalents of 1, 3-benzothiazole-2-amine and 0.1 equivalent of Pd₂(dba)₃A mixture of 0.2 equivalents XantPhos and 3 equivalents DIPEA in CyOH (5mL/mmol) was maintained at 140 ℃. After the appropriate conversion was reached, the reaction mixture was diluted with DCM (10mL/mmol), injected onto a pretreated silica gel column and purified by flash chromatography using heptane/EtOAc as eluent.

Buchwald general procedure II

A mixture of chloro compound suspended in 1, 4-dioxane (5mL/mmol), 2 equivalents of 1, 3-benzothiazol-2-amine, 10 mol% JosiPhos Pd (G3), and 3 equivalents of DIPE was stirred at reflux until no further conversion was observed. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography on a 120g silica gel column using heptane-EtOAc or DCM-MeOH (1.2% NH)₃) As an eluent.

Mitsunobu general method

To a mixture of 1 equivalent of aliphatic alcohol, 1 equivalent of carbamate/phenol and 1 equivalent of triphenylphosphine in toluene (5mL/mmol) was added 1 equivalent of di-tert-butyl azodicarboxylate. The mixture is stirred at 50 ℃ for carbamates or at room temperature for phenols. After the appropriate conversion was reached, the volatiles were removed under reduced pressure and the crude intermediate was purified by flash chromatography using heptane/EtOAc as eluent.

Finkelstein universal method

A mixture of 1 equivalent of alkyl chloride and 2 equivalents of NaI in acetone (5mL/mmol) was maintained at reflux. After the appropriate conversion was reached, the volatiles were removed under reduced pressure and the crude intermediate was purified by flash chromatography using heptane/EtOAc as eluent.

General procedure for quaternary salt formation

In an oven-dried vial was equipped with a PTFE-coated magnetic stir bar, 1 equivalent of tosylate was added and 20 equivalents of the appropriate amine were suspended in CyOH (5 mL/mmol). The reaction mixture was then warmed to 140 ℃ and stirred at this temperature until no further conversion was observed. The reaction mixture was diluted with DCM and then poured onto a silica gel column pretreated with DCM. Then purified by flash chromatography using DCM and MeOH (1.2% NH ₃) As an eluent.

Universal deprotection method for quaternary salts

To a solution of the appropriate quaternary salt in THF (5mL/mmol) was added 3 equivalents of TBAF followed by stirring at room temperature until no further conversion was observed. The reaction mixture was evaporated to dryness under reduced pressure. To a suspension of 1 equivalent of the desalted quaternary salt in anhydrous MeCN (15mL/mmol) was added 100 equivalents of HF x Pyr, followed by stirring at 60 ℃. After the appropriate conversion was reached, the volatiles were removed under reduced pressure, the residue was suspended in a 1:1 mixture of THF-water (30mL/mmol), and 150 equivalents of LiOH x H were added₂O, the mixture was stirred at room temperature. After the appropriate conversion was reached, the volatiles were removed under reduced pressure. The crude product was purified by flash chromatography using DCM and MeOH (containing 1.2% NH₃) As an eluent.

General process for the preparation of propynylamine

In an oven-dried vial was equipped with a PTFE-coated magnetic stir bar and 2 equivalents of PPh were added₃And 2 equivalents of imidazole, then DCM (5mL/mmol) was added. To the resulting mixture was added 2 equivalents of iodine in portions, followed by stirring at room temperature for 15 minutes. To the resulting mixture was added 1 equivalent of the appropriate alcohol dissolved in DCM and stirred at room temperature until no further conversion was observed. To the resulting iodo compound was added 20 equivalents of the appropriate amine, followed by stirring at room temperature for 30 minutes while complete conversion was observed. Adding diatomaceous earth to the reaction mixture Of the reaction mixture, volatiles were removed under reduced pressure. Then purified by flash chromatography using DCM and MeOH (1.2% NH₃) As an eluent.

General method for preparing propargylamine catalyzed by silver

In a 24ml vial was equipped with a stir bar and 1 equivalent of 2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] was added]Pyridazin-8-yl radicals]-5- [3- (4-ethynyl-2-fluoro-phenoxy) propyl]Thiazole-4-carboxylic acid, 20 equivalents paraformaldehyde/acetone and 20 equivalents of the appropriate amine were stirred in anhydrous ethanol (5ml/mmol) in the presence of 20 mol% silver tosylate at 80 ℃ until no further conversion was observed. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. Then purified by flash chromatography using DCM and MeOH (1.2% NH₃) As an eluent.

General method of hydrolysis

The appropriate methyl ester was suspended in a 1:1 mixture of THF-water (5mL/mmol) and 10 equivalents of LiOH x H were added₂O, the mixture was stirred at 50 ℃. After reaching a suitable conversion, the volatiles were removed under reduced pressure; the crude product was purified by flash chromatography using DCM and MeOH (containing 1.2% NH₃) As an eluent.

General procedure for amine substitution and hydrolysis

To a solution of any of the products of preparations 12, 13 and 14 in a 1:1 mixture (10ml/mmol) of acetonitrile and N-methyl-2-pyrrolidone was added the appropriate amine (3-10 equivalents), the reaction mixture was stirred at 50 ℃ for 2-24H, after purification of the substituted product by column chromatography (silica gel using DCM and MeOH as eluent), the product was dissolved in THF (10ml/mmol), water (2ml/mmol) and LiOH XH were added ₂O (3-5 equivalents). Then, the reaction mixture was stirred at 20-40 ℃ for 1-4 hours. The hydrolysate was purified by preparative HPLC (using acetonitrile and 5mM NH)₄HCO₃Aqueous solution as eluent) to afford the desired product.

Preparation examples

The following experimental details describe the preparation of synthetic intermediates.

Preparation example 1a2- (tert-butoxycarbonylamino) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid methyl ester

Step A2- (tert-Butoxycarbonylamino) -5-iodo-thiazole-4-carboxylic acid methyl ester

50.00g of methyl 2- (tert-butoxycarbonylamino) thiazole-4-carboxylate (193.55mmol,1eq) were suspended in 600mL of dry MeCN. 52.25g N-iodosuccinimide (232.30 mmol) was added and the resulting mixture was stirred at room temperature overnight.

The reaction mixture was diluted with saturated brine and then extracted with EtOAc. The combined organic layers were washed with 1M Na₂S₂O₃Extracted and then extracted again with brine. Then using Na₂SO₄Dried, filtered and the filtrate concentrated under reduced pressure. The crude product was purified by flash chromatography using heptane as eluent to give 60g of the desired product (156mmol, 80% yield).

¹H NMR(400MHz,DMSO-d₆):δppm 12.03/11.06(br s),3.78(s,3H),1.47(s,9H)；¹³C NMR(100MHz,DMSO-d₆)δppm 153.8,82.5,77.7,52.3,28.3；HRMS-ESI(m/z):[M+H]⁺C₁₀H₁₄IN₂O₄384.9713 as the calculated value of S; found 384.9708.

Step B2- (tert-Butoxycarbonylamino) -5- (3-hydroxyprop-1-ynyl) thiazole-4-carboxylic acid methyl ester

A500 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar and fitted with a reflux condenser. To this was added 9.6g of the product of step A (25mmol,1eq), 2.80g of prop-2-yn-1-ol (2.91mL,50mmol,2eq) and 36.10g of DIPA (50mL,356.8mmol,14.27eq), followed by 125mL of dry THF and the system was flushed with argon. After stirring for 5 minutes under an inert atmosphere 549mg Pd (PPh3) were added₂Cl₂(1.25mmol,0.05eq) and 238mg CuI (1.25mmol,0.05 eq). The resulting mixture was then warmed to 60 ℃ and stirred at that temperature until no further conversion was observed. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. Then purifying by flash chromatographyConversion using heptane and EtOAc as eluent gave 7.30g of the desired product as a yellow solid (23mmol, 93% yield).

¹H NMR(400MHz,DMSO-d₆):δppm 12.1(br s,1H),5.45(t,1H),4.36(d,2H),3.79(s,3H),1.48(s,9H)；¹³C NMR(100MHz,DMSO-d₆)δppm 12.1(br s,1H),5.45(t,1H),4.36(d,2H),3.79(s,3H),1.48(s,9H)；HRMS-ESI(m/z):[M+H]⁺C₁₃H₁₇N₂O₅Calculated value of S is 313.0852, found value is 313.0866.

Step C2- (tert-Butoxycarbonylamino) -5- (3-hydroxypropyl) thiazole-4-carboxylic acid methyl ester

A1L oven-dried pressurized bottle was equipped with a PTFE-coated magnetic stir bar to which was added 44.75g of the product of step B (143.3mmol,1eq), 7.62Pd/C (7.17mmol,0.05eq) in 340mL of ethanol, then placed under a nitrogen atmosphere using a hydrogenation system. Then 4 bar H ₂Filled and stirred at room temperature overnight. Complete conversion is observed, but only olefin product is formed. After filtering the catalyst through a pad of celite, the entire process was repeated with 5 mol% fresh catalyst. The resulting mixture was stirred overnight until complete conversion was achieved. Celite was added to the reaction mixture and the volatiles were removed under reduced pressure. It was then purified by flash column chromatography using heptane and EtOAc as eluent to give 31.9g of the desired product as pale yellow crystals (101mmol, 70.4% yield).

¹H NMR(500MHz,DMSO-d₆):δppm 11.61(br s,1H),4.54(t,1H),3.76(s,3H),3.43(m,2H),3.09(t,2H),1.74(m,2H),1.46(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.8,143.1,135.4,60.3,51.9,34.5,28.3,23.4；HRMS-ESI(m/z):[M+H]⁺C₁₃H₂₁N₂O₅Calculated value of S is 317.1165, found value is 317.1164(M + H).

Step D2- (tert-butoxycarbonylamino) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid methyl ester

In a 250mL oven-dried single-necked round bottom flask equipped with a PTFE-coated magnetic stir bar, 3.40g of 2-fluoro-4-iodo-phenol was added(14mmol,1eq), 5.00g of the product of step C (16mmol,1.1eq) and 4.10g of PPh dissolved in 71mL of dry toluene₃(16mmol,1.1 eq). After stirring for 5 minutes under nitrogen, 3.10mL of DIAD (3.20g,16mmol,1.1eq) was added in one portion while the reaction mixture was heated. The reaction mixture was then heated to 50 ℃ and stirred at this temperature for 30 minutes, the reaction giving complete conversion.

The reaction mixture was injected directly onto a pre-treated silica gel column, which was then purified by flash chromatography using heptane and EtOAc as eluents. The crude product was crystallized from MeOH to give 4.64g of the desired product (9.24mmol, 66% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 11.64(br s,1H),7.59(dd,1H),7.45(dd,1H),6.98(t,1H),4.06(t,2H),3.73(s,3H),3.22(t,2H),2.06(m,2H),1.46(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 134,124.9,117.6,68.2,51.9,30.5,28.3,23.2；HRMS-ESI(m/z):[M+H]⁺C₁₉H₂₃N₂O₅Calculated value of FSI 537.0350; found 537.0348.

Preparation example 1b2- (tert-butoxycarbonylamino) -5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A500 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar and fitted with a reflux condenser. To this were added 13.41g of preparation 1a (25mmol,1eq), 8.46g N-methyl-N-prop-2-ynyl-carbamic acid tert-butyl ester (50mmol,2eq) and 50mL of DIPA (36.10g,50mL,356.8mmol,14.27eq), followed by 125mL of dry THF and the system flushed with argon. After stirring for 5 minutes under an inert atmosphere 549mg Pd (PPh) were added₃)₂Cl₂(1.25mmol,0.05eq) and 238mg CuI (1.25mmol,0.05 eq). The resulting mixture was then warmed to 60 ℃ and stirred at that temperature until no further conversion was observed. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc as eluent to give 10.5g of the desired product (18.2mmol, 72.7% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 11.65(br s,1H),7.31(br d,1H),7.21(br d,1H),7.14(t,1H),4.23(s,2H),4.1(t,2H),3.73(s,3H),3.23(t,2H),2.86(s,3H),2.07(m,2H),1.46/1.41(s,18H)；¹³C NMR(125MHz,DMSO-d₆)δppm 129.1,119.2,115.4,68.1,51.9,38.6,33.8,30.5,23.2；HRMS-ESI(m/z):[M+H]⁺C₂₈H₃₇FN₃O₇578.2330 as the calculated value of S; found 578.2331.

Preparation example 1c2- (tert-butoxycarbonylamino) -5- [3- [4- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A250 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar and fitted with a reflux condenser. To this were added 5.36g of preparation 1a (10mmol,1eq), 1.66g N, N-dimethylprop-2-yn-1-amine (20mmol,2eq) and 20mL of DIPA (142.7mmol,14.27eq), followed by 50mL of dry THF and the system was flushed with argon. After stirring for 5 minutes under an inert atmosphere, 220mg of Pd (PPh) were added₃)₂Cl₂(0.5mmol,0.05eq) and 95CuI (0.5mmol,0.05 eq). The resulting mixture was then warmed to 60 ℃ and stirred at that temperature until no further conversion was observed. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using DCM and MeOH (1.2% NH₃) As eluent 4.5g of the desired product (7.8mmol, 78% yield) were obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 11.66(s,1H),7.29(dd,1H),7.19(m,1H),7.12(t,1H),4.09(t,2H),3.73(s,3H),3.44(s,2H),3.23(t,2H),2.24(s,6H),2.07(m,2H),1.45(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.8,147.3,129,119.2,115.4,84.3,68,51.9,48.1,44.2,30.6,28.3,23.2；HRMS-ESI(m/z):[M+H]⁺C₂₄H₃₁FN₃O₅492.1962 as the calculated value of S; found 492.1956(M + H).

Preparation example 1d2- { [ (tert-butoxy) carbonyl]Amino } -5- (3-iodopropyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 1a, step C (5g,15.8mmol,1eq) in diethyl ether (175mL) and acetonitrile (35mL) was added imidazole (1.57mL,23.71mmol,1.5eq), followed by triphenylphosphine (3.73g,14.22mmol,1.5eq) and iodine (6.02g,23.71mmol,1.5 eq). The mixture was stirred at room temperature for 1 hour. The reaction was partitioned between ethyl acetate (150mL) and 10% aqueous sodium thiosulfate (250mL) and the organic phase was washed successively with water (200mL) and brine (150mL), dried (magnesium sulfate) and concentrated in vacuo. The residue was dissolved in ether and aged at refrigerator temperature overnight. The crystals formed were removed by filtration and the filtrate was concentrated in vacuo. Purification by automated flash chromatography (Combiflash Rf, Silica 80g RediSep column) with a gradient elution of 0-50% ethyl acetate in isoheptane afforded the desired product as a white solid (5.77g,13.53mmol, 85%).

LC/MS(C₁₃H₁₉IN₂O₄S)427[M+H]⁺；RT 0.88(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ11.67(s,1H),3.79(s,3H),3.29(t,J＝6.8Hz,2H),3.20-3.12(m,2H),2.09(dq,J＝8.7,6.8Hz,2H),1.48(s,9H)。

Preparation example 2a3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propan-1-ol

Step A[ (pent-4-yn-1-yloxy) methyl group]Benzene and its derivatives

To an oven dried flask was added a solution of 4-pentyn-1-ol (11.1mL,119mmol,1eq) in THF (100mL) and the solution was cooled to 0 ℃. Sodium hydride (60% dispersion; 7.13g,178mmol,1.5eq) was added portionwise and the mixture was stirred at 0 ℃ for 30 minutes, then benzyl bromide (15.6mL,131mmol,1.1eq) was added dropwise. The mixture was warmed to room temperature and stirred for 16 hours, then cooled to 0 ℃, quenched with saturated aqueous ammonium chloride (30mL) and diluted with water (30 mL). The mixture was extracted with ethyl acetate (2 × 150mL) and the combined organic extracts were washed successively with dilute ammonium hydroxide solution (150mL) and brine (100mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,330 gReddiSep)^TMSilica gel cartridge) and gradient eluted with 0-10% ethyl acetate in isoheptane to give the desired product as a yellow liquid (19.5g,112mmol, 94%).

LC/MS(C₁₂H₁₄O)175[M+H]⁺；RT 1.28(LCMS-V-B1)

¹H NMR(400MHz,Chloroform-d)δ7.37-7.32(m,4H),7.31-7.27(m,1H),4.52(s,2H),3.58(t,J＝6.1Hz,2H),2.32(td,J＝7.1,2.6Hz,2H),1.95(t,J＝2.7Hz,1H),1.83(tt,J＝7.1,6.2Hz,2H)。

Step B[ (hex-4-yn-1-yloxy) methyl group]Benzene and its derivatives

To an oven dried flask, the product of step A (19.5g,112mmol,1eq) and tetrahydrofuran (200mL) were added and the solution was cooled to-78 ℃. N-butyllithium (66.9mL,135mmol,1.2eq) was added dropwise over 30 minutes and the reaction was stirred for 1 hour, then iodomethane (10.5mL,168mmol,1.5eq) was added dropwise and the mixture was warmed to 0 ℃ over 1 hour. The reaction was quenched by the addition of saturated aqueous ammonium chloride (40mL), diluted with water (40mL), extracted with ethyl acetate (3 × 100mL) and the combined organic extracts washed successively with 2M aqueous sodium thiosulfate (200mL) and brine (200mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,330g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-10% ethyl acetate in isoheptane to give the desired product as a yellow liquid (19.2g,0.1mol, 91%). LC/MS (C)₁₃H₁₆O)189[M+H]⁺；RT 1.34(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.41-7.23(m,5H),4.46(s,2H),3.48(t,J＝6.3Hz,2H),2.23-2.14(m,2H),1.72(s,3H),1.70-1.65(m,2H)。

Step C4- [3- (benzyloxy) propyl group]-3, 6-dichloro-5-methylpyridazine

A solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (5g,33.1mmol,1eq) and the product of step B (7.48g,39.8mmol,1.2eq) in tetrahydrofuran (30mL) was heated at 160 ℃ for 19 hours in a sealed flask. The reaction was cooled to room temperature and then concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as an orange oil (7.32g,23.5mmol, 71%).

LC/MS(C₁₅H₁₆Cl₂N₂O)311[M+H]⁺；RT 1.35(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.45-7.18(m,5H),4.48(s,2H),3.53(t,J＝5.9Hz,2H),2.96-2.83(m,2H),2.42(s,3H),1.88-1.69(m,2H)。

Step D3- (3, 6-dichloro-5-methylpyridazin-4-yl) propan-1-ol

To a cooled solution of the product of step C (7.32g,23.5mmol,1eq) in dichloromethane (100mL) was added dropwise a solution of boron trichloride (1M in dichloromethane; 58.8mL,58.8mmol,2.5eq) and the mixture was stirred at room temperature for 1 hour. The reaction was stopped by adding methanol and concentrated in vacuo. The residue was partitioned between dichloromethane (100mL) and saturated aqueous sodium bicarbonate (150mL) and the organic phase was washed with brine (150mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-80% ethyl acetate in isoheptane to give the desired product as a yellow oil (4.19g,19mmol, 81%).

LC/MS(C₈H₁₀Cl₂N₂O)221[M+H]⁺；RT 0.84(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ4.67(t,J＝5.1Hz,1H),3.49(td,J＝6.0,5.1Hz,2H),2.91-2.80(m,2H),2.43(s,3H),1.72-1.59(m,2H)。

Preparation example 2b2- [ tert-butyl (diphenyl) silyl group]Oxy-3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propan-1-ol (enantiomerically pure, enantiomer 2 from step A)

Step A3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) -2-hydroxy-propionic acid ethyl ester

To a solution of 3, 6-dichloro-4, 5-dimethyl-pyridazine (26.5g,150mmol) in dry THF (375mL) at-78 ℃ TMP-MgCl LiCl (165mL,165mmol,1.1eq.) was added dropwise, and the resulting mixture was stirred at 0 ℃ for 2 hours. The resulting magnesium salt was transferred to a solution of ethyl 2-oxoacetate (45.9g,225mmol,1.5eq.) in dry THF (375mL) at 0 ℃ and then stirred at 0 ℃ for 30 minutes. With saturated NH₄After the reaction was quenched with aqueous Cl and extracted with EtOAc, the combined organic layers were dried, filtered, concentrated and purified by flash silica gel chromatography,using heptane and EtOAc as eluent gave 11g (26.3%) of the desired compound.

¹H NMR(500MHz,DMSO-d₆)δppm 5.85(d,1H),4.33(m,1H),4.12(q,2H),3.19(d,2H),2.45(s,3H),1.17(t,3H)；¹³C NMR(125MHz,DMSO-d₆)δppm 172.8,157.6,157.2,141.4,139.3,68.8,61.2,35.2,17.3,14.4.HRMS-ESI(m/z):[M+H]+C₁₀H₁₃Cl₂N₂O₃279.0303, found 279.0301.

The enantiomers of the desired product were separated on an AS-V chiral column (100 x 500mm,20 μm) using 10:90 EtOH-heptane AS eluent to give enantiomer 1 in 99.6% ee (eluted first) and enantiomer 2 in 99.1% ee (eluted later).

And B, step B:2- [ tert-butyl (diphenyl) silyl ] silane]Oxy-3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propionic acid ethyl ester

To a solution of enantiomer 2(4500mg,16mmol) of step a, imidazole (2200mg,2.0eq.) in THF (81mL) was added TBDPS-Cl (8900mg,2.0eq.) dropwise, followed by stirring at room temperature for 18 hours. The product was purified by flash chromatography using heptane and EtOAc as eluents to give the desired product (6200mg, 74%).

¹H NMR(400MHz,DMSO-d₆)δppm 7.52-7.27(m,10H),4.46(dd,1H),3.83(m,2H),3.35(dd,1H),3.19(dd,1H),2.34(s,3H),0.93(t,3H),0.87(s,9H)。

Step C2- [ tert-butyl (diphenyl) silyl group]Oxy-3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propan-1-ol

To a solution of the enantiomerically pure product of step B (3600mg,6.95mmol) in MeOH (35mL) at 0 ℃ in 5 min was added NaBH in portions₄(2.63g,10eq.) and stirred at this temperature for 30 minutes. By adding saturated NH₄After the reaction was quenched with aqueous Cl, extracted twice with EtOAc. The combined organic layers were dried, filtered, concentrated and purified by flash silica gel chromatography using heptane and EtOAc as eluent to give the desired product (1.6g, 48%).

¹H NMR(500MHz,DMSO-d₆)δppm 7.57-7.3(m,10H),4.9(brs,1H),4.05(m,1H),3.38/3.32(dd+dd,2H),3.13/3.11(dd+dd,2H),2.3(s,3H),0.8(s,9H)；¹³C NMR(125MHz,DMSO-d₆)ppm 72.7,65.5,35.5,26.9,17.2；HRMS-ESI(m/z):[M+H]⁺C₂₄H₂₉Cl₂N₂O₂Calculated Si value 475.1369, found 475.1362.

Preparation example 2c2- (3, 6-dichloro-5-methyl-pyridazin-4-yl) ethanol

Step A3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propane-1, 2-diol

To a solution of 700mg (2.5mmol) of the product of preparation 2b, step A in 3mL of methanol at 0 deg.C was added 285mg (3eq.) of NaBH ₄And the mixture was stirred at 0 ℃ for 0.5 hour. With saturated NH₄After the reaction was stopped with Cl solution, the crude product was purified by flash chromatography on silica gel with DCM and MeOH (1.2% NH₃) As eluent 500mg (84%) of the desired compound were obtained.

¹H NMR(400MHz,DMSO-d₆)δppm 4.90(bd,1H),4.83(bs,1H),3.75(m,1H),3.47(dd,1H),3.38(m,1H),3.00(dd,1H),2.87(dd,1H),2.45(s,3H)。

Step B2- (3, 6-dichloro-5-methyl-pyridazin-4-yl) acetaldehyde

237mg of the product of step A (1mmol.) in 5mL of acetone/H₂The O (4:1) solution was cooled to 0 ℃ and 427mg of sodium periodate (2mmol,2eq.) were added portionwise. After stirring at room temperature for 2 h, the mixture was purified by flash chromatography using heptane-EtOAc as eluent to give 200mg of the desired product (97%).

¹H NMR(400MHz,DMSO-d₆)δppm 9.71(s,1H),4.27(s,1H),2.35(s,3H)。

Step C2- (3, 6-dichloro-5-methyl-pyridazin-4-yl) ethanol

To a solution of 200mg of the product of step B (0.97mmol) in 3mL of methanol at 0 deg.C were added in small portions 110mg (2.92mmol,3eq.) of sodium borohydride. After stirring for 15 minutes, the reaction mixture was taken up with saturated NH₄Aqueous Cl was diluted and extracted with EtOAc. The combined organic layers were dried, filtered, concentrated and purified by flash chromatography using heptane-EtOAc as eluentThe dose gave 180mg (89%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 4.9(t,1H),3.65(m,2H),3(t,2H),2.45(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δppm 157.5,157.2,140.9,140.7,59.1,34,17.1；HRMS-ESI(m/z):[M+H]+C₇H₉Cl₂N₂Calculated value of O207.0086, found value 207.0083.

Preparation example 2e3- (3, 6-dichloro-5-methylpyridazin-4-yl) propanal

To an oven dried flask was added dimethyl sulfoxide (3.08mL,43.4mmol,2.4eq) and dichloromethane (100mL) and the solution was cooled to-78 ℃. Oxalyl chloride (2M in dichloromethane; 13.6mL,27.1mmol,1.5eq) was added dropwise and the reaction was stirred for 1 hour. The product of preparation 2a (4g,18.1mmol,1eq) was then added dropwise in dichloromethane (20mL) and the mixture was stirred for 1 h. Triethylamine (15.1mL,109mmol,6eq) was added and the reaction was warmed to 0 ℃ over 1 hour. The reaction was quenched with water (50mL), then partitioned between saturated sodium bicarbonate (50mL) and dichloromethane (200mL), the aqueous phase was extracted with dichloromethane (200mL), the combined organic extracts were washed with brine (100mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to afford the desired product as an off-white solid (2.27g,10.4mmol, 57%).

LC/MS(C₈H₈Cl₂N₂O)219[M+H]⁺；RT 0.87(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ9.71(s,1H),3.03(dd,J＝8.7,7.0Hz,2H),2.86-2.69(m,2H),2.44(s,3H)。

Preparation example 3a2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid methyl ester

Step A2- { [ (tert-butoxy) carbonyl][3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl group]Amino } -5- [3- (2-fluoro-4-iodophenoxy) propyl]-1, 3-thiazole-4-carboxylic acid methyl ester

Using the general method of Mitsunobu, starting with 4.85g of preparation 1a (9.04mmol,1eq) as the appropriate carbamate and 2g of preparation 2a (9.04mmol,1eq) as the appropriate alcohol, 4.6g of the desired product was obtained (69% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.56(dd,1H),7.44(dm,1H),7.08(m,2H),6.96(t,1H),4.05(t,2H),3.75(s,3H),3.21(t,2H),2.82(m,2H),2.4(s,3H),2.06(m,2H),1.88(m,2H),1.48(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.7,157.6,156.7,156.5/153.2,152.2,147,142.1,139.8,134,124.9,117.6,84,82.4,68.1,52.1,46.1,30.4,28.1,27.5,25.8,23.1,16.4；HRMS-ESI(m/z):[M+H]⁺C₂₇H₃₁Cl₂FIN₄O₅Calculated value of S is 739.0415, found value is 739.0395.

Step B2- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino group]-5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for deprotection with HFIPA, starting from the product of step A as the appropriate carbamate, 3.70g of the desired product was obtained (97% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.71(t,1H),7.59(dd,1H),7.44(dm,1H),6.96(t,1H),4.03(t,2H),3.7(s,3H),3.29(m,2H),3.11(t,2H),2.84(m,2H),2.39(s,3H),2(m,2H),1.76(m,2H)；¹³C NMR(125MHz,DMSO-d₆)δppm 164.6,163,152.3,147.1,134.1,124.8,117.6,82.4,68.1,51.9,44,30.7,28,26.9,23.3,16.4；HRMS-ESI(m/z):[M+H]⁺C₂₂H₂₃Cl₂FIN₄O₃Calculated value of S is 638.9891, found value is 638.9888.

Step C2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl ]Thiazole-4-carboxylic acid methyl ester

A suspension of 3g of the product of step B (4.69mmol,1eq) and 1.81g of cesium carbonate (9.3853mmol,2eq.) in 25mL of dry 1, 4-dioxane was stirred at 80 ℃ for 3 hours to reach complete conversion. The reaction mixture was directly evaporated onto celite and then purified by flash chromatography using DCM-MeOH as eluent to give 2.67g of the title compound (94% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.57(dd,1H),7.43(dm,1H),6.97(t,1H),4.23(t,2H),4.08(t,2H),3.77(s,3H),3.22(t,2H),2.86(t,2H),2.29(s,3H),2.08(m,2H),2.03(m,2H)；

¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,155.4,152.2,151.6,151.2,147,142.5,136,134.8,134,128.9,124.9,117.6,82.3,68.4,51.9,46.3,30.7,24.2,23,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₂₂H₂₂ClFIN₄O₃Calculated value of S603.0124, found value 603.0108.

Preparation example 3b5- (3-hydroxypropyl) -2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Step A2- (tert-butoxycarbonylamino) -5- [3- [ tert-butyl (dimethyl) silyl]Oxyprop-1-ynyl]Thiazole-4-carboxylic acid methyl ester

In a 1L oven dried single neck round bottom flask equipped with a PTFE-coated magnetic stir bar, 20g of preparation 1a, step a (52.05mmol,1.0eq.) was added to it, 17.73g of tert-butyl-dimethyl-prop-2-ynyloxy-silane (21mL,104.1mmol,2.0eq.) dissolved in 250mL of dry THF/25mL of DIPA, then placed under nitrogen by passing nitrogen through the gas inlet. To this solution was added 572mg Pd (PPh)₃)₂Cl₂(1.30mmol,0.025eq.) and 247mg of CuI (1.30mmol,0.025 eq.). The reaction mixture was then warmed to reflux and stirred at that temperature until no further conversion was observed. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography in two portions using heptane and EtOAc as eluent to give 18.00g of the desired product (81% yield).

¹H NMR(400MHz,DMSO-d₆)δppm 12.13(br.,1H),4.62(s,2H),3.79(s,3H),1.48(s,9H),0.89(s,9H),0.13(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 161.2,52.4,52.4,28.3,26.2,-4.6；HRMS-ESI(m/z):[M+H]⁺C₁₉H₃₁N₂O₅Calculated value of SSi 427.1717, found value 427.1711.

Step B2- (tert-butoxycarbonylamino) -5- [3- [ tert-butyl (dimethyl) silyl]Oxopropyl radical]Thiazole-4-carboxylic acid methyl ester

13g of the product of step A (30.42mmol,1.0eq.) was dissolved in 150mL EtOH and 3.23g Pd/C (3.04mmol,0.1eq.) was added. A 250mL oven-dried autoclave was equipped with a PTFE-coated magnetic stir bar, to which the solution was added, and then placed under a nitrogen atmosphere using a hydrogenation system. Then it was treated with 10 bar H₂And (6) filling. After stirring at room temperature for 2 hours, the reaction was completely converted. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc as eluent to give 9.95g of the desired product (78% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 11.62(br.,1H),3.76(s,3H),3.62(t,2H),3.12(t,2H),1.78(quint.,2H),1.46(s,9H),0.86(s,9H),0.03(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm162.8,62,51.9,34.3,28.3,26.3,23.3,-4.9；HRMS-ESI(m/z):[M+H]⁺C₁₉H₃₅N₂O₅Calculated value of SSi 431.2030, found value 431.2025.

Step C2- [ tert-Butoxycarbonyl- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propyl ] oxy]Amino group]-5- [3- [ tert-butyl (dimethyl) silyl]Oxopropyl radical]Thiazole-4-carboxylic acid methyl ester

Using the Mitsunobu general procedure, starting with 9.91g of the product of step B (23.0mmol,1eq.) as the appropriate carbamate and 5.1g of preparation 2a (23.0mmol,1eq) as the appropriate alcohol, 13.02g of the desired product was obtained (89% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 4.09(t,2H),3.77(s,3H),3.61(t,2H),3.12(t,2H),2.82(t,2H),2.41(s,3H),1.88(qn,2H),1.79(qn,2H),1.39(s,9H),0.85(s,9H),0.02(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.8,157.7,156.3,156.1,152.8,144.5,142.1,139.9,135.3,79.4,62.1,52.1,46.1,34.1,28.6,27.5,26.3,25.9,23.2,18.4,16.4,-4.9；HRMS-ESI(m/z):[M+H]⁺C₂₇H₄₃Cl₂N₄O₅Calculated value of SSi 633.2095, found value of 633.2091.

Step D5- [3- [ tert-butyl (dimethyl) silyl ] group]Oxopropyl radical]-2- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for deprotection with HFIPA, starting from the product of step C as the appropriate carbamate, 10.4g of the desired product was obtained (95% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.69(t,1H),3.71(s,3H),3.60(t,2H),3.30(q,2H),3.01(t,2H),2.85(t,2H),2.41(s,3H),1.78(qn,2H),1.71(qn,2H),0.86(s,9H),0.02(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 164.3,163.1,157.7,156.9,142.5,140.0,137.6,136.5,62.0,51.7,44.1,34.4,28.0,26.9,26.3,23.4,18.5,16.5,-4.9；HRMS-ESI(m/z):[M+H]⁺C₂₂H₃₅Cl₂N₄O₃Calculated value of SSi 533.1570, found value of 533.1566.

Step E5- [3- [ tert-butyl (dimethyl) silyl group]Oxopropyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

A250 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar, to which was added 10.4g of the product of step D (19.57mmol,1.0eq.), 12.75g Cs₂CO₃(39.13mmol,2.0eq.) and 100mL dry 1, 4-dioxane. The reaction mixture was then warmed to reflux temperature and stirred at that temperature for 8 hours. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc as eluent to give 6.40g of the desired product (66% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 4.26(t,2H),3.79(s,3H),3.65(t,2H),3.14(t,2H),2.89(t,2H),2.32(s,3H),2.04(m,2H),1.82(m,2H),0.87(s,9H),0.04(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,155.3,151.8,151.3,143.4,136.1,134.6,129.0,62.1,52.0,46.3,34.4,26.3,24.2,23.1,19.7,15.7,-4.8；HRMS-ESI(m/z):[M+H]⁺C₂₂H₃₄ClN₄O₃Calculated value of SSi 497.1804, found value 497.1796.

Step F2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] ]Pyridazin-8-yl radicals]-5- [3- [ tert-butyl (dimethyl) silyl]Oxopropyl radical]Thiazole-4-carboxylic acid methyl ester

A250 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar and fitted with a reflux condenser. To this were added 6.43g of the product of step E (12.94mmol,1.0eq.), 3.88g of 1, 3-benzothiazol-2-amine (25.87mmol,2.0eq) and 6.75mL of DIPEA (38.81mmol,3.0eq.), followed by 65mL of CyOH. The system was then flushed with argon. Stirring for 5 minutes under an inert atmosphere and then adding 1.18g Pd₂(dba)₃(1.29mmol,0.1eq.) and 1.49g XantPhos (2.587mmol,0.2 eq.). The resulting mixture was then warmed to 140 ℃ and stirred at this temperature for 1 hour to achieve complete conversion. The reaction mixture was diluted with DCM and injected directly onto a pre-treated silica gel column, which was then purified by flash chromatography using heptane and EtOAc as eluent to give 6.85g of the desired product (87% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.82(br.,1H),7.52(br.,1H),7.37(t,1H),7.19(t,1H),4.25(t,2H),3.80(s,3H),3.66(t,2H),3.16(t,2H),2.87(t,2H),2.33(s,3H),2.04(m,2H),1.84(m,2H),0.92(s,9H),0.07(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.2,155.6,148.8,148.6,142.3,134.5,127.6,126.5,122.5,122,62.0,51.9,46.3,34.4,26.4,23.9,22.9,20.3,12.8,-4.8；HRMS-ESI(m/z):[M+H]⁺C₂₉H₃₉N₆O₃S₂Calculated Si value 611.2288, found 611.2284.

Step G5- [3- [ tert-butyl (dimethyl) silyl group]Oxopropyl radical]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

5.00g of the product of step F (8.18mmol,1.0eq.) was dissolved in 50mL of dry DCM and 50mg of DMAP (0.41mmol,0.05eq.) and 2.85mL of DIPEA (16.37mmol,2.0eq.) were added at 0 ℃. Then 2.24mL 2- (chloromethoxy) ethyl-trimethyl-silane (12.69mmol,1.5eq.) were added over 5 minutes at 0 ℃ and the resulting mixture was cooled in a refrigerator overnight while complete conversion was observed. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc as eluent to give 3.85g of the desired product (63% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.6-7.15(m,4H),5.83(s,2H),4.42(t,2H),3.92(s,3H),3.74(t,2H),3.73(t,2H),3.24(t,2H),2.86(t,2H),2.37(s,3H),2.12(m,2H),1.97(m,2H),0.96(t,2H),0.95(s,9H),0.1(s,6H),-0.07(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.6,157.7,156.4,154.7,148.5,143.7,137.6,134.1,132.6,126.1,125.6,73.2,66.9,62.5,51.9,46,34.3,26.1,24.2,23.4,20.6,18.0,12.9,-1.4,-5.2；HRMS-ESI(m/z):[M+H]⁺C₃₅H₅₃N₆O₄S₂Si₂741.3102, found 741.3098.

Step H5- (3-hydroxypropyl) -2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

3.85G of the product of step G (5.19mmol,1.0eq.) and 362mg camphorsulfonic acid (1.56mmol,0.3eq.) were dissolved in 40mL DCM/MeOH (2: 1). The reaction mixture was then warmed to 50 ℃ and stirred at this temperature overnight. The reaction was completely converted. The reaction mixture was cooled to room temperature and purified by addition of saturated NaHCO₃The reaction was quenched with aqueous solution and then extracted twice with EtOAc. Celite was added to the combined organic layers and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc as eluent to give 2.50g of the title compound (76% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.83(dm,1H),7.44(dm,1H),7.42(m,1H),7.23(m,1H),5.84(s,2H),4.57(brs,1H),4.26(t,2H),3.80(s,3H),3.72(m,2H),3.48(t,2H),3.14(m,2H),2.86(t,2H),2.36(s,3H),2.04(m,2H),1.81(m,2H),0.91(m,2H),-0.11(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 127.1,123.3,123.2,111.9,72.9,66.7,60.6,51.9,46.4,35.0,23.8,23.2,20.4,17.8,13,-1.0；HRMS-ESI(m/z):[M+H]+C₂₉H₃₉N₆O₄S₂Calculated Si 627.2237, found 627.2236.

Preparation example 3c2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- (4-ethynyl-2-fluoro-phenoxy) propyl]Thiazole-4-carboxylic acid

Step A2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- [ 2-fluoro-4- (2-trimethylsilylethynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A250 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar and fitted with a reflux condenser. To this was added 5g of preparation 3a (8.29mmol,1eq.), 2.34mL of ethynyl (trimethyl) silane (16.58mmol,2eq.) and 10mL of DIPEA, followed by addition of 40mL of dry THF and flushing of the system with argon. After stirring for 5 minutes under an inert atmosphere, 182mg Pd (PPh) was added₃)₂Cl₂(0.41mmol,0.05eq.) and 79mg (0.41mmol,0.05 eq.). The resulting mixture was then warmed to 60 ℃ and stirred at this temperature for 2 hours to achieve complete conversion. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane-EtOAc as eluent to give 4.26g of the desired product (89% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.31(dd,1H),7.23(dn,1H),7.13(t,1H),4.25(t,2H),4.12(t,2H),3.77(s,3H),3.24(t,2H),2.87(t,2H),2.31(s,3H),2.1(m,2H),2.03(m,2H),0.21(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.0,155.3,151.7,151.3,136.1,129.4,129.0,119.4,115.3,104.6,93.7,68.2,51.9,46.3,30.7,24.1,23.0,19.7,15.7,0.4；HRMS-ESI(m/z):[M]+C₂₇H₃₀ClFN₄O₃Calculated value of SSi 572.1481, found value 572.1480.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (2-trimethylsilylethynyl) phenoxy group]Propyl radical]Thiazole-4-carboxylic acid methyl ester

In a 100mL oven dried single neck round bottom flask was fitted a PTFE-coated magnetic stir bar to which were added 4.25g of the product of step a (7.4mmol,1.0eq.), 2.23g of 1, 3-benzothiazol-2-amine (14.8mmol,2.0eq.) and 3.87mL of DIPEA (2.87mg,22.2mmol,3.0eq.), then 40mL of cyclohexanol was added and the system was flushed with argon. Stirring for 5 minutes under inert atmosphere and then adding 679mg Pd₂(dba)₃(0.74mmol,0.10eq.) and 858mg XantPhos (1.48mmol,0.20 eq.). The resulting mixture was then warmed to 140 ℃ and stirred at this temperature for 30 minutes to achieve complete conversion. The reaction mixture was diluted with DCM and injected directly onto a pre-treated silica gel column, which was then purified by flash chromatography using heptane and EtOAc as eluent. The pure fractions were combined and concentrated under reduced pressure to give 3.90g of the desired product (77% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 12.27/10.91(brs,1H),8.1-7.1(brm,4H),7.34(dd,1H),7.24(dm,1H),7.16(t,1H),4.25(t,2H),4.15(t,2H),3.78(s,3H),3.28(t,2H),2.87(t,2H),2.34(s,3H),2.13(m,2H),2.04(m,2H),0.19(s,9H)；HRMS-ESI(m/z):[M+H]+C₃₄H₃₆FN₆O₃S₂Calculated Si value 687.2038, found 687.2020.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals ]-5- [3- (4-ethynyl-2-fluoro-phenoxy) propyl]Thiazole-4-carboxylic acid

A10 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar and fitted with a reflux condenser. Thereto was added a solution of 2.5mL THF/H₂343mg of O (4:1) product of step B (0.5mmol,1.0 eq.). Then 105mg of LiOH x H were added₂O (2.50mmol,5.0eq.) and the resulting mixture was heated to 60 ℃ and stirred at this temperature for 4 hours. The reaction was completely converted. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. Then passing it through the tunnelPurification by chromatography using DCM and MeOH (1.2% NH)₃) As eluent, 200mg of the title compound (66% yield) were obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.88(d,1H),7.49(br.,1H),7.37(t,1H),7.36(dd,1H),7.25(dm,1H),7.19(t,1H),7.16(t,1H),4.27(t,2H),4.15(t,2H),4.11(s,1H),3.27(t,2H),2.87(t,2H),2.33(s,3H),2.14(m,2H),2.04(m,2H)；¹³C NMR(125MHz,DMSO-d₆)δppm 164.2,151.5,147.9,129.4,126.5,122.5,122.3,119.5,115.5,114.5,82.9,80.5,68.5,46.2,31.0,23.9,23.1,20.3,12.9；HRMS-ESI(m/z):[M+H]+C₃₀H₂₆FN₆O₃S₂601.1486, found 601.1498.

Preparation example 3d2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-hydroxyprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Step A5- [3- [4- [3- [ tert-butyl (dimethyl) silyl group]Oxyprop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

Using the Sonogashira general procedure, starting with 4.00g of preparation 3a (6.63mmol,1.0eq.) and 2.26g of tert-butyl-dimethyl-prop-2-ynyloxy-silane (13.27mmol,2eq.) as the appropriate alkyne, 2.80g of the desired product was obtained (65% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.27(dd,1H),7.19(dd,1H),7.14(t,1H),4.51(s,1H),4.25(m,2H),4.12(t,2H),3.77(s,3H),3.24(t,2H),2.87(t,2H),2.3(s,3H),2.1(quint.,2H),2.03(m,2H),0.88(s,9H),0.12(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.0,128.9,119.1,115.5,68.3,52.1,51.9,46.3,30.7,26.2,24.2,23.0,19.7,15.7,-4.6；HRMS-ESI(m/z):[M+H]+C₃₁H₃₉ClFN₄O₄Calculated value of SSi 645.2128, found value of 645.2120.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ tert-butyl (dimethyl)Radical) silane radical]Oxyprop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure II, starting with 2.8g of the product of step A (4.34mmol,1.0eq.) and 1.30g of 1, 3-benzothiazol-2-amine (8.67mmol,2.0eq.) gives 2.1g of the desired product (64% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 12.25/10.91(brs 1H),7.88(br,1H),7.51(br,1H),7.37(t,1H),7.29(dd,1H),7.2(t,1H),7.2(dd,1H),7.17(t,1H),4.49(s,2H),4.25(t,2H),4.14(t,2H),3.77(s,3H),3.27(t,2H),2.86(t,2H),2.32(s,3H),2.13(qn,2H),2.04(qn,2H),0.87(s,9H),0.1(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.2,155.7,151.6,148.5,147.6,141.5,128.9,127.6,126.5,122.5,122.3,119.1,116.9,115.5,114.8,88.2,84,68.4,52.1,51.9,46.4,31,26.2,24,23.1,20.4,12.9,-4.6；HRMS-ESI(m/z):[M+H]+C₃₈H₄₄FN₆O₄S₂Calculated Si value 759.2613, found 759.2609.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-hydroxyprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A PTFE-coated magnetic stir bar was fitted into a 100mL oven-dried single-necked round bottom flask and a reflux condenser was installed. To this was added 2.10g of the product of step B (2.76mmol,1.0eq.) dissolved in 15mL of THF. Then 3.32mL of TBAF (3.32mmol,1.2eq, 1M in THF) was added dropwise via syringe over 2 minutes and stirred at this temperature for 30 minutes. The reaction mixture was washed with saturated NH₄The reaction was quenched with Cl, then evaporated directly onto celite and purified by flash chromatography using heptane-EtOAc as eluent to give 1.6g of the desired product (90% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 11.14(brs,1H),7.83(brd,1H),7.49(brs,1H),7.36(m,1H),7.24(dd,1H),7.19(m,1H),7.18(dm,1H),7.15(t,1H),5.08(t,1H),4.28(m,2H),4.27(d,2H),4.17(t,2H),3.8(s,3H),3.29(m,2H),2.89(m,2H),2.35(s,3H),2.15(m,2H),2.07(m,2H)；HRMS-ESI(m/z):[M+H]+C₃₂H₃₀FN₆O₄S₂645.1748, found 645.1738.

Preparation example 3e2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-hydroxypropyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Step A2- (tert-butoxycarbonylamino) -5- [3- [4- [3- [ tert-butyl (dimethyl) silyl]Oxyprop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the Sonogashira general procedure, starting with 4.00g of preparation 1a (7.45mmol,1.0eq.) and 2.54g of tert-butyl-dimethyl-prop-2-ynyloxy-silane (14.90mmol,2.0eq.) as the appropriate alkyne, 1.70g of the desired product was obtained (39% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 11.64(s,1H),7.27(dd,1H),7.19(dm,1H),7.14(t,1H),4.51(s,2H),4.1(t,2H),3.73(s,3H),3.23(t,2H),2.07(m,2H),1.46(s,9H),0.89(s,9H),0.12(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 88.2,83.8。

Step B2- (tert-butoxycarbonylamino) -5- [3- [4- [3- [ tert-butyl (dimethyl) silyl]Oxopropyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A PTFE-coated magnetic stir bar was assembled in a 50mL oven-dried autoclave. To this were added 1.70g of the product of step A (2.9mmol,1.0eq.), 310mg Pd/C (0.29mmol,0.10eq.) and 15mL of ethanol, then inertized with vacuum and nitrogen and finally filled with hydrogen at 10 bar pressure. The mixture was then stirred at room temperature for 3 hours to reach complete conversion. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc as eluent to give 1.2g of the desired product (70% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 11.64(br.,1H),7.02(t,1H),7.01(d,1H),6.89(d,1H),4.02(t,2H),3.74(s,3H),3.54(t,2H),3.22(t,2H),2.54(t,2H),2.04(quint.,2H),1.70(quint.,2H),1.45(s,9H),0.85(s,9H),0(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.8,156.2/153.5,152.0,144.7,141.9,135.8,135.5,124.6,116.2,115.5,68.1,62.0,51.9,34.3,30.8,30.8,28.3,26.2,23.2,-4.9。

Step C2- [ tert-Butoxycarbonyl- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propyl ] oxy]Amino group]-5- [3- [4- [3- [ tert-butyl (dimethyl) silyl group]Oxopropyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the Mitsunobu general procedure, starting with 1.16g of the product of step B (2.0mmol,1.0eq.) as the appropriate carbamate and 484mg of preparation 2a (2.2mmol,1.1eq.) as the appropriate alcohol, 1.2g of the desired product was obtained (77% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.02(m,1H),6.99(d,1H),6.89(m,1H),4.08(t,2H),4.02(t,2H),3.75(s,3H),3.54(t,2H),3.22(t,2H),2.81(t,2H),2.53(t,2H),2.40(s,3H),2.05(quint.,2H),1.87(m,2H),1.70(quint.,2H),1.48(s,9H),0.85(s,9H),0.00(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.7,156.4/153,152.0,144.7,143.6,142/139.8,141.9,135.5,124.6,116.2,115.4,68.1,62.0,52.0,46.1,34.2,30.8,30.7,28.0,27.5,26.2,25.8,23.2,16.4,-4.9；

Step D5- [3- [4- [3- [ tert-butyl (dimethyl) silyl group]Oxopropyl radical]-2-fluoro-phenoxy]Propyl radical]-2- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for deprotection with HFIPA, starting with 1.2g of the product of step C as the appropriate carbamate, 790mg of the desired product was obtained (75% yield).

Step E5- [3- [4- [3- [ tert-butyl (dimethyl) silyl group]Oxopropyl radical]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

A25 mL oven-dried pressurized bottle was equipped with a PTFE-coated magnetic stir bar to which were added 1.2g of the product of step D (1.75mmol,1.0eq) and 680mg of cesium carbonate (3.50mmol,2.0eq) suspended in 10mL of 1, 4-dioxane. The reaction mixture was then warmed to 80 ℃ and stirred at this temperature for 3 hours, at which time the reaction reached full conversion. Adding to the reaction mixture Celite and the volatiles were removed under reduced pressure. It was then purified by flash chromatography using DCM and MeOH (with 1.2% NH)₃) As eluent, 1.0g of the desired product was obtained (88% yield).

Step F2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ tert-butyl (dimethyl) silyl group]Oxopropyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure II, using 630mg of the product of step E (0.97mmol,1.0eq.) and 291mg of 1, 3-benzothiazol-2-amine (1.94mmol,2.0eq.) as starting material, 600mg of the desired product (81%) was obtained.

Step G2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-hydroxypropyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A PTFE-coated magnetic stir bar was fitted into a 250mL oven-dried round bottom flask. 600mg of the product of step F (0.78mmol,1.0eq.) dissolved in 10mL of THF was added to it, followed by dropwise addition of 936uL of TBAF (0.963mmol,1.2 eq.). Complete conversion was observed after 1 hour of stirring. The reaction mixture was then washed with saturated NH₄The reaction was terminated with aqueous Cl, celite was added to the reaction mixture and the volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc and MeOH (1.2% NH) ₃) As eluent 450mg of the desired product was obtained (89% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.87(br,1H),7.49(br,1H),7.37(t,1H),7.19(t,1H),7.06(m,1H),7.05(d,1H),6.92(dd,1H),4.44(br,1H),4.25(t,2H),4.08(t,2H),3.78(s,3H),3.36(t,2H),3.27(t,2H),2.85(t,2H),2.52(t,2H),2.32(s,3H),2.1(qn,2H),2.04(qn,2H),1.65(qn,2H)；¹³C NMR(500MHz,dmso-d6)δppm 163.2,155.6,152.0,148.5,144.7,141.7,135.9,134.8,127.6,126.5,124.7,122.5,122.3,116.3,116.0,115.6,68.6,60.4,52.0,46.4,34.6,31.2,31.0,23.9,23.2,20.4,12.9。

Preparation example 3f2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazol-4-yl radicals-Carboxylic acid ethyl ester

Step A2- [ (hex-4-yn-1-yl) amino]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of ethyl 2-bromo-1, 3-thiazole-4-carboxylate (1.17g,4.97mmol,1eq) in acetonitrile (16mL) was added hex-4-yn-1-amine (725mg,7.46mmol,1.5eq) and triethylamine (1.04mL,7.46mmol,1.5eq) and the mixture was heated under microwave irradiation at 150 ℃ for 4 hours. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to afford the desired product as a beige solid (741mg,2.94mmol, 59%).

LC/MS(C₁₂H₁₆N₂O₂S)253[M+H]⁺；RT 2.32(LCMS-V-C)

Step B2- { 3-chloro-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (443mg,2.94mmol,1eq) in tetrahydrofuran (15mL) was added the product of step A (741mg,2.94mmol,1eq) and the mixture was heated in a sealed tube at 110 ℃ overnight. The reaction was concentrated in vacuo and the residue triturated with methanol, filtered and dried in vacuo to afford the desired product as a beige solid (607mg,1.79mmol, 61%).

LC/MS(C₁₄H₁₅ClN₄O₂S)339[M+H]⁺；RT 2.41(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.06(s,1H),4.38-4.25(m,4H),2.92(t,J＝6.3Hz,2H),2.34(s,3H),2.14-2.01(m,2H),1.31(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step B (607mg,1.79mmol,1eq), 2-aminobenzothiazole (404mg,2.69mmol,1.5eq)), XantPhos (207mg,0.36mmol,0.2eq), cesium carbonate (1.17g,3.58mmol,2eq) and 1, 4-dioxane (36mL) and the mixture was stirredThe vessel was evacuated and flushed with nitrogen, then tris (dibenzylideneacetone) dipalladium (0) (164mg,0.18mmol,0.1eq) was added and the mixture was purged with nitrogen (10min) and then heated at 150 ℃ for 4 hours under microwave irradiation. The reaction was diluted with ethyl acetate and filtered through celite, then washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give a solid which was triturated with ether, filtered and dried in vacuo to give the desired product as a yellow solid (329mg,0.73mmol, 41%).

LC/MS(C₂₁H₂₀N₆O₂S₂)453[M+H]⁺；RT 2.73(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.99(br s+s,2H),7.65(br s,1H),7.43-7.31(m,1H),7.28-7.15(m,1H),4.35-4.25(m,4H),2.96-2.85(m,2H),2.36(s,3H),2.15-2.00(m,2H),1.32(t,J＝7.1Hz,3H)。

Preparation example 3g5- (3-hydroxypropyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene ]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

Step A2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of preparation 3f (11.7g,25.8mmol,1eq) in dimethylformamide (700mL) was added N, N-diisopropylethylamine (13.5mL,77.4mmol,3 eq). After 5 min the mixture was cooled to 0 ℃ and 4- (dimethylamino) pyridine (630mg,5.16mmol,0.2eq) and 2- (trimethylsilyl) ethoxymethyl chloride (13.6mL,77.4mmol,3eq) were added and the mixture was stirred at room temperature overnight. The reaction was concentrated in vacuo, then partitioned between dichloromethane and brine, the organic phase dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,330g RediSep)^TMSilica gel cartridge) purification with 0-40% ethyl acetate in isoheptaneElution afforded the desired product as a yellow solid (9.61g,16.5mmol, 64%).

LC/MS(C₂₇H₃₄N₆O₃SiS₂)583[M+H]⁺；RT 2.90(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.99(s,1H),7.82(dd,J＝7.7,1.1Hz,1H),7.49-7.38(m,2H),7.28-7.19(m,1H),5.86(s,2H),4.38-4.23(m,4H),3.77-3.67(m,2H),2.89(t,J＝6.2Hz,2H),2.38(s,3H),2.13-2.01(m,2H),1.31(t,J＝7.1Hz,3H),0.91(dd,J＝8.5,7.4Hz,2H),-0.11(s,9H)。

Step B5-bromo-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c) ]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (9.61g,16.5mmol,1eq) in dichloromethane (400mL) was added N-bromosuccinimide (3.52g,19.8mmol,1.2eq) and the mixture was stirred at room temperature overnight. The reaction was partitioned between dichloromethane and water and the organic phase was washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to afford the desired product as a yellow solid (9.66g,14.6mmol, 89%).

LC/MS(C₂₇H₃₃BrN₆O₃SiS₂)663[M+H]⁺；RT 3.13(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.84(dd,J＝7.5,1.1Hz,1H),7.59-7.38(m,2H),7.24(ddd,J＝8.3,6.7,1.7Hz,1H),5.85(s,2H),4.37-4.23(m,4H),3.72(dd,J＝8.5,7.4Hz,2H),2.87(t,J＝6.2Hz,2H),2.38(s,3H),2.13-2.00(m,2H),1.32(t,3H),0.95-0.81(m,2H),-0.12(s,9H)。

Step C5- [ (1E) -3- [ (tert-butyldimethylsilyl) oxy group]Prop-1-en-1-yl]-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried sealed flask was added the product of step B (9.66g,14.6mmol,1eq), (E) -3- (tert-butyldimethylsilyloxy) propen-1-yl-boronic acid pinacol ester (5.74mL,17.5mmol,1.2eq), potassium carbonate (6.05g,43.8mmol,3eq), [1,1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride (1.19g,1.46mmol,0.1eq), tetrahydrofuran (360mL) and water (120mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ for 2 h. The reaction was partitioned between ethyl acetate and water, and the organic layer was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a yellow solid (6.46g,8.58mmol, 59%).

LC/MS(C₃₆H₅₂N₆O₄Si₂S₂)753[M+H]⁺；RT 1.62(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.80(dd,J＝7.6,1.0Hz,1H),7.51-7.38(m,3H),7.24(ddd,J＝8.3,6.8,1.8Hz,1H),6.28(dt,J＝16.0,4.3Hz,1H),5.85(s,2H),4.37(dd,J＝4.4,2.1Hz,2H),4.35-4.25(m,4H),3.72(dd,J＝8.5,7.4Hz,2H),2.88(t,J＝6.3Hz,2H),2.37(s,3H),2.09-1.99(m,2H),1.31(t,J＝7.1Hz,3H),0.93(s,9H),0.92-0.83(m,2H),0.11((s,6H),-0.11(s,9H)。

Step D5- {3- [ (tert-butyldimethylsilyl) oxy ] group]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step C (6.46g,8.58mmol,1eq) in ethyl acetate (300mL) under nitrogen was added platinum (IV) oxide (390mg,1.72mmol,0.2 eq). The vessel was evacuated and backfilled with nitrogen (x3), then evacuated, placed under a hydrogen atmosphere and shaken at room temperature for 3 days. The reaction was filtered through celite, eluting with ethyl acetate and concentrated in vacuo to afford the desired product as a brown gum (6.72g,8.9mmol, > 100%).

LC/MS(C₃₆H₅₄N₆O₄Si₂S₂)755[M+H]⁺；RT 1.67(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.76(d,1H),7.48-7.35(m,2H),7.24(ddd,J＝8.2,6.5,1.9Hz,1H),5.84(s,2H),4.33-4.22(m,4H),3.76-3.62(m,4H),3.15(t,J＝7.5Hz,2H),2.87(t,J＝6.4Hz,2H),2.37(s,3H),2.10-1.98(m,3H),1.91-1.79(m,2H),1.31(t,J＝7.1Hz,3H),0.95-0.85(m,11H),0.06(s,6H),-0.12(s,9H)。

Step E5- (3-hydroxypropyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step D (6.72g,8.9mmol,1eq) in 1, 4-dioxane (400mL) was added hydrochloric acid (4M in dioxane; 67mL,267mmol,30eq) and the mixture was stirred at room temperature for 1 h. The reaction was cooled to 0 ℃ and neutralized with 1N aqueous sodium hydroxide (300mL) then partitioned between ethyl acetate and water, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-80% ethyl acetate in isoheptane to give a solid which was triturated with ether, filtered and dried in vacuo to give the desired product as a white solid (3.87g,6.04mmol, 68%).

LC/MS(C₃₀H₄₀N₆O₄SiS₂)641[M+H]⁺；RT 2.80(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.83(dd,J＝7.6,1.1Hz,1H),7.48-7.37(m,2H),7.23(ddd,J＝8.3,6.7,1.8Hz,1H),5.85(s,2H),4.56(t,J＝5.1Hz,1H),4.33-4.22(m,4H),3.72(dd,J＝8.6,7.3Hz,2H),3.48(td,J＝6.3,5.1Hz,2H),3.17-3.08(m,2H),2.88(t,J＝6.4Hz,2H),2.38(s,3H),2.11-1.99(m,2H),1.87-1.75(m,2H),1.31(t,J＝7.1Hz,3H),0.96-0.86(m,2H),-0.11(s,9H)。

Preparation example 4a4- [1- [ (dimethylamino) methyl group]-3-bicyclo [1.1.1]Pentyl radical]-2-fluoro-phenol

Step ATricyclic [1.1.1.0 ]¹,³]Pentane (pentane)

A1L 3-necked flask equipped with a stir bar was equipped with a distillation head connected to a condenser and a 250mL collection flask with a Schlenk tap, a 250mL dropping funnel and a thermometer [ all glassware was heat-soaked assembled, then connected to a vacuum line and cooled under a stream of nitrogen]. A solution of 1, 1-dibromo-2, 2-bis (chloromethyl) cyclopropane (59.4g,200mmol,1eq) in diethyl ether (200mL) was cooled to-45 ℃ and phenyllithium (1.9M solution in n-butyl ether; 211mL,400mmol,2eq) was added over 25 minutes via the dropping funnel. After the addition was complete, the mixture was allowed to warm to 0 ℃ and stirred for 2 hours. Thereafter, the receiving flask was cooled to-78 ℃, the connection to the manifold was briefly closed and replaced with a vacuum pump attachment (pressure equalization inlet connected to nitrogen manifold). The dropping funnel and thermometer were replaced by a pre-greased glass stopper before the pump was turned on. The pump was brought to a pressure of 200 mbar and the connection was then opened. The pressure was gradually reduced to 120 mbar within 3 minutes and then the reaction vessel was allowed to warm to ambient temperature. The pressure was then carefully reduced to 45 mbar and maintained for 45 minutes. Thereafter, the vacuum was released with nitrogen and the resulting clear colorless distillate was stored at-20 ℃. By passing ¹H NMR determined the concentration of the desired product to be 0.45M.

¹H NMR (400MHz, chloroform-d) delta 2.04(s, 6H).

Step BBromine (3-fluoro-4-methoxyphenyl) magnesium

A3-neck 50mL flask was equipped with a stir bar and condenser, to which was added magnesium (681mg,28mmol,1.4eq) and the apparatus was heated vigorously (. about.500 ℃) with a heat gun for 5 minutes under vigorous stirring, then cooled to room temperature under nitrogen. Diethyl ether (5mL) was added followed by 1, 2-dibromoethane (172. mu.L, 2mmol,0.1 eq). The mixture was heated to reflux 4-5 times over 5 minutes and then allowed to stand for 10 minutes, after which a gentle reflux was observed. The mixture was heated by hand to steady reflux and then slow stirring was started. At this point a solution of 4-bromo-2-fluoroanisole (4.1g,20mmol,1eq) in ether (10mL) was added at a rate sufficient to maintain stable reflux and the stirring rate (300rpm) was increased. The addition was complete after 15 minutes. The mixture was stirred at room temperature for 0.5 h, then a clear two-phase system was obtained. The lower, dark straw layer (10.15mL) was transferred by syringe through a 0.2um PTFE filter into a dry Schlenk flask. The concentration of the solution was calculated to be 1.38M by titration of a solution of iodine in anhydrous tetrahydrofuran. The product solution was used in the next step without further characterization.

Step C3- (3-fluoro-4-methoxyphenyl) bicyclo [1.1.1]Pentane-1-carboxylic acid ethyl ester

A stir bar was equipped in an oven-dried 50mL ACE pressurized container to which the product of step B (1.38M in ether; 4.83mL,6.67mmol,1eq) was added, then the product of step A (0.45M in ether, 14.8mL,6.67mmol,1eq) was added and the container was sealed with a Teflon screw cap fitted with a front O-ring and then placed in a preheated heating block at 105 ℃ behind an explosion proof hood for 3 hours. The mixture was cooled at room temperature for 20 minutes and then in ice water for 10 minutes. The teflon screw cap was replaced with subaseal connected to a nitrogen line and the reaction solution was cooled to-78 ℃. Ethyl chloroformate (5.1mL,53.3mmol,4eq) was added and the mixture was warmed to room temperature for 1.5 h. The reaction was partitioned between saturated aqueous ammonium chloride and diethyl ether and the aqueous phase was extracted with diethyl ether. The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% ethyl acetate in isoheptane to give the desired product as a colorless liquid as a mixture of the desired product and by-products. The product was further subjected to automatic flash column chromatography (CombiFlash Torrent,200g RediSep) ^TMSilica gel cartridge) and eluted with a gradient of 0-80% dichloromethane in heptane to give the desired product (640mg,3.78mmol, 56%).

¹H NMR(500MHz,DMSO-d6)δppm 7.09(t,1H),7.09(dd,1H),6.98(dm,1H),4.08(q,2H),3.8(s,3H),2.22(s,6H),1.2(t,3H)。13C NMR(500MHz,dmso-d6)δppm 169.8,151.8,146.6,133.0,122.8,114.2,114.2,60.6,56.5,53.2,41.0,36.9,14.6。

HRMS-EI(m/z):M+C₁₅H₁₇FO₃264.1162, found 264.1156.

Step D1- (3-fluoro-4-methoxy-phenyl) bicyclo [1.1.1]Pentane-3-carboxylic acid

200mg of the product of step C (0.76mmol,1eq.) and 159mg of LiOH XH₂O (3.78mmol,5eq.) was mixed in 1, 4-dioxane (2mL/mmol) and water (2mL/mmol) and then stirred at room temperature for 1 hour to observe complete conversion. The reaction mixture was basified with 1:1HCl solution, then the precipitate was filtered and washed with water, then dried under vacuum overnight. 170mg (95%) of the desired product are isolated as a white solid.

¹H NMR(500MHz,DMSO-d₆)δppm 12.41(s,1H),7.09(m,1H),7.09(m,1H),6.97(dm,1H),3.80(s,3H),2.18(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 171.7,151.8,146.6,133.3,122.7,114.2,114.1,56.5,53.1,40.8,37.0；GC-MS-EI(m/z):[M]⁺C₁₃H₁₃FO₃236.0849, found 236.0840.

Step E1- (3-fluoro-4-methoxy-phenyl) -N, N-dimethyl-bicyclo [1.1.1]Pentane-3-carboxamide

164mg of the product of step D (1.04mmol,1eq.) and 278mg of N, N-diethylethylamine (1.39mmol,2eq.) were mixed in EtOAc (3mL/mmol) and 663mg of 2,4, 6-tripropyl-1, 3,5, 2. lambda. were added in one portion⁵,4λ⁵,6λ⁵Trioxatriphosphane 2,4, 6-trioxide (50 w% EtOAc solution, 1.04mmol,1.5eq.) and then stirred at room temperature for 40 min. After the reaction time was reached, 0.52mL of N-methyl methylamine (2M in MeOH, 1.04mmol,1.5eq.) was added and stirred at room temperature until complete conversion was observed (60 min). The reaction mixture was then diluted with DCM and concentrated NaHCO ₃Washing, then washing the organic phase with concentrated NaCl, MgSO₄Drying, filtration, concentration and vacuum drying gave 187mg (quantitative) of the desired product as a pink solid.

¹H NMR(500MHz,DMSO-d₆)δppm 7.14(m,2H),6.86(m,2H),3.72(s,3H),3.08(s,3H),2.81(s,3H),2.26(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 168.9,158.6,132.5,127.6,114.1,55.5,54.2,42.0,39.0,37.4,35.9；HRMS-ESI(m/z):[M+H]⁺C₁₅H₁₉FNO₂264.1394, found 264.1389.

Step F1- [3- (3-fluoro-4-methoxy-phenyl) -1-bicyclo [1.1.1]Pentyl radical]-N, N-dimethyl-methylamine

182mg of the product of step E (0.69mmol,1eq.) was dissolved in THF (5mL/mmol) and then 1.38mL of LiAlH was added under nitrogen at room temperature₄(1M solution in THF, 1.38mmol,2eq.) and then stirred until complete conversion is achieved (about 1 hour). The mixture was cooled to 0 ℃ and then concentrated NH was used₄The reaction was stopped with Cl. After the reaction was terminated, 5mL of water and 10mL of EtOAc were added and shaken well. 2M HCl was added and the (acidic) aqueous phase was separated off, and the organic phase was then further extracted with 2M HCl. The combined aqueous phases were basified with 2M NaOH and extracted with DCM. The combined organic phases were washed with brine, dried over magnesium sulfate and concentrated, dried in vacuo. 119mg (69%) of the desired product are obtained as a viscous oil.

¹H NMR(500MHz,DMSO-d₆)δppm 7.07(t,1H),7.01(dd,1H),6.93(dm,1H),3.79(s,3H),2.35(s,2H),2.16(s,6H),1.90(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 151.8,146.2,134.5,122.5,114.1,114.0,60.7,56.5,52.9,46.6,41.7,38.0；HRMS-ESI(m/z):[M+H]⁺C₁₅H₂₁Calculated FNO 250.1602, found 250.1596.

Step G4- [1- [ (dimethylamino) methyl group]-3-bicyclo [1.1.1]Pentyl radical]-2-fluoro-phenol

113mg of the product of step F (0.45mmol,1eq.) was dissolved in DCM (5mL/mmol) and 1.36mL of BBr was added at 0 deg.C under nitrogen ₃(1M solution in DCM, 1.36mmol,3eq.) and then stirred at 0 ℃ for 15 minutes and then at room temperature until complete conversion is achieved (about 45 min). DCM was added and then NaHCO was poured in₃In solution, stirred for a few minutes, then concentrated NH is used₄And (4) neutralizing with Cl. Separated and washed with brine, dried over magnesium sulfate and concentrated, dried under vacuum. 47mg (quantitative) of the crude desired product are obtained as a viscous oil.

¹H NMR(400MHz,CDCl₃)δppm 6.95(t,1H),6.90(dd,1H),6.85(dm,1H),3.84(s,2H),3.17(s,6H),2.24(s,6H)；¹³C NMR(100MHz,CDCl₃)δppm 122.4,117.4,113.4,59.5,54.8,46.0,43.8,34.8；HRMS-ESI(m/z):[M+H]⁺C₁₄H₁₉Calculated value of FNO 236.1445,236.1445.

Preparation example 4b4- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenol

Using the Sonogashira general procedure, starting with 10.00g of 2-fluoro-4-iodo-phenol (42.0mmol,1eq.) as the appropriate phenol and 5.24g N, N-dimethylprop-2-yn-1-amine (63mmol,1.5eq.) as the alkyne, 7.30g (90%) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.20(dd,1H),7.07(dm,1H),6.91(m,1H),3.39(m,2H),2.21(m,3H)；¹³C NMR(125MHz,DMSO-d₆)δppm 150.9,146.2,128.9,119.5,118.4,113.6,84.5,84.2,48.2,44.3；HRMS-ESI(m/z):[M+H]⁺C₁₁H₁₃Calculated FNO 194.0976, found 194.0981.

Preparation example 4cN- [3- (3-fluoro-4-hydroxy-phenyl) prop-2-ynyl]-N-methyl-carbamic acid tert-butyl ester

Using the Sonogashira general procedure, starting with 10.00g of 2-fluoro-4-iodo-phenol (42.0mmol,1eq.) as the appropriate phenol and 10.67g N-methyl-N-prop-2-ynyl-carbamic acid tert-butyl ester (63.1mmol,1.5eq.) as the alkyne, 10.8g (92%) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 10.32(s,1H),7.22(brd,1H),7.08(dm,1H),6.92(dd,1H),4.21(s,2H),2.85(s,3H),1.41(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 150.8,146.4,129.0,119.6,118.4,113.2,84.4,82.7,38.5,33.8,28.5；HRMS-ESI(m/z):[M-C₄H₈+H]⁺C₁₁H₁₁FNO₃224.0717, found 224.0720.

Preparation example 4d4- [3- (dimethylamino) propyl group]-2-fluorophenol

To a solution of the product of preparation 4b (1.5g,7.76mmol,1eq) in ethyl acetate (54mL) and ethanol (18mL) under nitrogen was added platinum (IV) oxide hydrate (353mg,1.55mmol,0.2 eq).The vessel was evacuated and backfilled with nitrogen (x3), then evacuated, hydrogenated and shaken at room temperature overnight. The reaction was filtered through celite, eluted with ethyl acetate and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% 1N methanolic ammonia in dichloromethane afforded the desired product as an off white solid (652mg,3.31mmol, 42%).

LC/MS(C₁₁H₁₆FNO)198[M+H]⁺；RT 0.44(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ9.52(s,1H),6.96(dd,J＝12.5,1.9Hz,1H),6.88-6.76(m,2H),2.47(dd,J＝8.5,6.8Hz,2H),2.20-2.13(m,2H),2.11(s,6H),1.69-1.57(m,2H)。

Preparation example 4e4- [2- (dimethylamino) ethoxy]Phenol and its preparation

Step A4- (methoxymethoxy) phenol

To a solution of hydroquinone (0.76mL,9.08mmol,1eq) in acetone (30mL) was added potassium carbonate (2.51g,18.2mmol,2eq) and chloromethyl methyl ether (0.69mL,9.08mmol,1eq) and the mixture was stirred at room temperature overnight. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-20% ethyl acetate in isoheptane to give the desired product as a brown oil (601mg,3.9mmol, 43%).

¹H NMR(400MHz,DMSO-d6)δ9.03(s,1H),6.91-6.80(m,2H),6.72-6.62(m,2H),5.05(s,2H),3.36(s,3H)。

Step B{2- [4- (methoxymethoxy) phenoxy group]Ethyl dimethyl amine

To a solution of the product of step A (400mg,2.59mmol,1eq) in tetrahydrofuran (20mL) were added N, N-dimethylethanolamine (526. mu.L, 5.19mmol,2eq), di-tert-butyl azodicarboxylate (1.19g,5.19mmol,2eq) and triphenylphosphine (1.36g,5.19mmol,2eq) and the mixture was heated at 50 ℃ for 3 hours. The reaction was concentrated in vacuo, partitioned between dichloromethane and saturated aqueous sodium bicarbonate, and the organic phase was separated (PTFE phase separation)Vessel) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-7% methanol in dichloromethane afforded the desired product as a brown oil (383mg,1.7mmol, 66%).

LC/MS(C₁₂H₁₉NO₃)226[M+H]⁺；RT 0.88(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ6.99-6.90(m,2H),6.94-6.82(m,2H),5.10(s,2H),3.98(t,J＝5.8Hz,2H),3.36(s,3H),2.59(t,J＝5.9Hz,2H),2.20(s,6H)。

Step C4- [2- (dimethylamino) ethoxy]Phenol and its preparation

A solution of the product of step B (383mg,1.7mmol,1eq) in hydrochloric acid (4M in 1, 4-dioxane; 5mL,20mmol,11.7eq) was stirred at room temperature for 1 h. The reaction was concentrated in vacuo and then dissolved in methanol, applied to an SCX cartridge (10g) wetted with methanol, washed with methanol and eluted with 1.75N methanolic ammonia and concentrated in vacuo to afford the desired product as a brown solid (249mg,1.37mmol, 812%).

LC/MS(C₁₀H₁₅NO₂)182[M+H]⁺；RT 0.24(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.91(s,1H),6.79-6.70(m,2H),6.70-6.62(m,2H),3.92(t,J＝5.9Hz,2H),2.57(t,J＝5.9Hz,2H),2.20(s,6H)。

Preparation example 4f4- [2- (pyrrolidin-1-yl) ethoxy ]Phenol as the starting material

Step A1- {2- [4- (methoxymethoxy) phenoxy group]Ethyl pyrrolidine

To a solution of the product of preparation 4e, step A (525mg,3.41mmol,1eq) in tetrahydrofuran (20mL) was added 1- (2-hydroxyethyl) pyrrolidine (0.8mL,6.81mmol,2eq), di-tert-butyl azodicarboxylate (1.57g,6.81mmol,2eq) and triphenylphosphine (1.79g,6.81mmol,2eq) and the mixture was heated at 50 ℃ overnight. The reaction was concentrated in vacuo and partitioned between dichloromethane and saturated aqueous sodium bicarbonate, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) purification with 0-7% methanolA gradient elution of methyl chloride solution afforded the desired product as a brown oil (556mg,2.21mmol, 65%).

LC/MS(C₁₄H₂₁NO₃)252[M+H]⁺；RT 1.09(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ6.98-6.91(m,2H),6.91-6.82(m,2H),5.10(s,2H),4.00(t,J＝5.9Hz,2H),3.36(d,J＝6.0Hz,3H),2.75(t,J＝6.0Hz,2H),2.50-2.42(m,4H),1.74-1.61(m,4H)。

Step B4- [2- (pyrrolidin-1-yl) ethoxy]Phenol and its preparation

A solution of the product of step A (556mg,2.21mmol,1eq) in hydrochloric acid (4M in 1, 4-dioxane; 7mL,28mmol,12.7eq) was stirred at room temperature for 30 minutes. The reaction was concentrated in vacuo, then dissolved in methanol, loaded onto an SCX cartridge (10g) wetted with methanol, washed with methanol, eluted with 1.75N methanolic ammonia and concentrated in vacuo to afford the desired product as a brown solid (453mg,2.19mmol, 99%).

LC/MS(C₁₂H₁₇NO₂)208[M+H]⁺；RT 0.28(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.90(s,1H),6.79-6.70(m,2H),6.70-6.62(m,2H),3.94(t,J＝6.0Hz,2H),3.17(d,J＝4.3Hz,2H),2.73(t,J＝6.0Hz,2H),2.49(dt,J＝4.1,1.4Hz,2H),1.74-1.60(m,4H)。

Preparation example 4g4- [2- (dimethylamino) ethyl group]-2-fluorophenol

Step A2-fluoro-1-methoxy-4- [ (E) -2-nitrovinyl]Benzene and its derivatives

To a solution of 3-fluoro-4-methoxybenzaldehyde (400mg,2.6mmol,1eq) and nitromethane (339. mu.L, 6.23mmol,2.4eq) in methanol (50mL) cooled to 0 ℃ was added dropwise a 1M aqueous solution of sodium hydroxide (20mL,20mmol,7.71eq) and the resulting mixture was stirred at 0 ℃ for 1 hour. The mixture was added portionwise to 8M aqueous hydrochloric acid (12mL,96mmol,37eq) cooled to 0 ℃ and the resulting suspension was warmed to room temperature and stirred for 30 min. The precipitate was collected by filtration, washed with water and dried in vacuo to give the desired product as a yellow solid (393mg,1.99mmol, 76%).

¹H NMR(400MHz,DMSO-d6)δ8.20(d,J＝13.6Hz,1H),8.10(dd,J＝13.5,1.0Hz,1H),7.88(dd,J＝12.6,2.1Hz,1H),7.70(dt,J＝8.6,1.5Hz,1H),7.29(t,J＝8.8Hz,1H),3.92(s,3H)。

Step B2- (3-fluoro-4-methoxyphenyl) ethan-1-amine

To a solution of the product of step A (393mg,1.99mmol,1eq) in tetrahydrofuran (12mL) was added lithium aluminum hydride (1M in tetrahydrofuran; 5.98mL,5.98mmol,3eq) and the mixture was heated at 40 ℃ overnight. The reaction was quenched with water (1.2mL) and concentrated in vacuo. The residue was dissolved in 2N aqueous hydrochloric acid (20mL) and washed with ethyl acetate (× 2). Tartaric acid (2.1g) was added to the aqueous phase and the pH was adjusted to pH 11 with concentrated ammonium hydroxide. The mixture was extracted with dichloromethane (× 3), the combined organic extracts were separated (PTFE phase separator) and concentrated in vacuo to give the desired product as a yellow oil (252mg,1.49mmol, 75%).

LC/MS(C₉H₁₂FNO)170[M+H]⁺；RT 0.14(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.16-7.03(m,3H),3.80(s,3H),3.38-3.30(m,2H),2.79-2.69(m,2H),2.62-2.54(m,2H)。

Step C[2- (3-fluoro-4-methoxyphenyl) ethyl group]Dimethylamine

To a solution of the product of step B (252mg,1.49mmol,1eq) in methanol (5mL) was added formaldehyde (13.4M in water; 123. mu.L, 4.47mmol,3eq) followed by sodium triacetoxyborohydride (947mg,4.47mmol,3eq) and glacial acetic acid (0.05mL) and the mixture was stirred at room temperature for 1 h. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMCartridge silica) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow oil (92mg,0.47mmol, 31%).

LC/MS(C₁₁H₁₆FNO)198[M+H]⁺；RT 0.82(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.16-6.94(m,3H),3.80(s,3H),2.64(dd,J＝8.5,6.7Hz,2H),2.41(dd,J＝8.5,6.7Hz,2H),2.16(s,6H)。

Step D4- [2- (dimethylamino) ethyl group]-2-fluorophenol

To a solution of the product of step C (92mg,0.47mmol,1eq) in dichloromethane (3.5mL) cooled to 0 ℃ was added boron tribromide (1M in dichloromethane, 1.4mL,1.4mmol,3eq) and the mixture was stirred at room temperature overnight. The reaction was cooled to 0 ℃ and quenched with methanol, then concentrated in vacuo. The residue was dissolved in methanol and loaded onto an SCX cartridge (5g) wetted with methanol, washed with methanol, eluted with 1.75N methanolic ammonia and concentrated in vacuo to afford the desired product as a brown oil (41mg,0.22mmol, 48%).

LC/MS(C₁₀H₁₄FNO)184[M+H]⁺；RT 0.36(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ9.55(s,1H),7.03-6.95(m,1H),6.88-6.78(m,2H),2.59(dd,J＝8.7,6.7Hz,2H),2.38(dd,J＝8.6,6.7Hz,2H),2.15(s,6H)。

Preparation example 4h3- [ (dimethylamino) methyl group]-5-fluoro-1-methyl-1H-indol-6-ol

Step A6- (benzyloxy) -5-fluoro-1H-indole-2-carboxylic acid

To a stirring solution of methyl 6-benzyloxy-5-fluoro-1H-indole-2-carboxylate (2.5g,8.35mmol,1eq) in a mixture of tetrahydrofuran (25mL) and methanol (25mL) was added a solution of sodium hydroxide (4g,100mmol,12eq) in water (30mL) and the mixture was stirred for 2.5 hours. The reaction was cooled in ice water and acidified by slowly adding 2N aqueous hydrochloric acid (60mL) with stirring to form a precipitate. Water (80mL) was added and the mixture was stirred for 45 min, then the solid was collected by filtration, washed with water and dried in vacuo to give the desired product as an off-white solid (2.25g,7.89mmol, 94%).

LC/MS(C₁₆H₁₂FNO₃)284[M-H]^-；RT 1.16(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ12.85(s,1H),11.71(d,J＝2.3Hz,1H),7.54-7.39(m,5H),7.39-7.32(m,1H),7.10(dd,J＝7.4,0.8Hz,1H),7.01(dd,J＝2.2,0.8Hz,1H),5.19(s,2H)。

Step B6- (benzyloxy) -5-fluoro-1H-indole

A mixture of the product of step A (1.25g,4.38mmol,1eq) and diphenyl ether (60mL) was heated at 290 deg.C (external temperature) for 45 min. The reaction was cooled to room temperature, then diluted with heptane (180mL) and loaded under vacuum onto a pre-packed silica column (80g) wetted with hexane. Purification by automated flash chromatography (CombiFlash Rf, silica gel 80g RediSep column) with a gradient elution of 0-80% ethyl acetate in hexanes afforded the desired product as a beige solid (372mg,1.54mmol, 35%).

LC/MS(C₁₅H₁₂FNO)242[M+H]⁺；RT 1.26(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ11.00(s,1H),7.52-7.45(m,2H),7.49-7.37(m,2H),7.41-7.30(m,2H),7.25(t,J＝2.8Hz,1H),7.13(dd,J＝7.3,0.8Hz,1H),6.34(ddd,J＝3.0,2.0,0.8Hz,1H),5.18(s,2H)。

Step C6- (benzyloxy) -5-fluoro-1-methyl-1H-indole

To a stirred solution of the product of step B (365mg,1.51mmol,1eq) in dimethylformamide (10mL) cooled in an ice-water bath was added sodium hydride (60% dispersion; 72.6mg,3.03mmol,2eq) and the mixture was stirred for 15 min. Methyl iodide (0.11mL,1.82mmol,1.2eq) was added and the mixture was then warmed to room temperature and stirred for 1 hour. The reaction solution was cooled in ice, and then quenched by dropwise addition of a saturated aqueous ammonium chloride solution and slowly poured into stirring ice water (40mL) to form a precipitate. Ice (20mL) was added and stirred for 30 min, then the solid was collected by filtration, washed successively with ice-cold water (2x30mL) and hexane (2x10mL) and dried in vacuo to give the desired product as a beige solid (259mg,1.01mmol, 67%).

LC/MS(C₁₆H₁₄FNO)256[M+H]⁺；RT 1.36(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.55-7.48(m,2H),7.47-7.38(m,2H),7.40-7.29(m,3H),7.25(d,J＝3.1Hz,1H),6.33(dd,J＝3.1,0.8Hz,1H),5.21(s,2H),3.76(s,3H)。

Step D{ [6- (benzyloxy) -5-fluoro-1-methyl-1H-indol-3-yl]Methyl dimethyl amine

To a stirred mixture of 1, 4-dioxane (5mL) and glacial acetic acid (5mL) was added aqueous formaldehyde (37 wt%; 1.18mL,14.54mmol,14.6eq) followed by aqueous dimethylamine (40 wt%; 1.42mL,12.6mmol,12.6 eq). This solution (1.6mL) was added to a stirred solution of the product of step C (255mg,1mmol,1eq) in 1, 4-dioxane (1mL) and the mixture was stirred at room temperature for 4 h. The reaction was concentrated in vacuo, then 2N aqueous sodium hydroxide (4mL) was added and the resulting thick suspension was diluted with water (10mL), stirred and cooled in ice water for 15 min, then filtered, washed with water (× 3) and dried in vacuo to give the desired product as a cream solid (290mg,0.93mmol, 93%).

LC/MS(C₁₉H₂₁FN₂O)268[M+H-NHMe₂]⁺；RT 0.99(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.55-7.47(m,2H),7.47-7.38(m,2H),7.40-7.31(m,2H),7.28(d,J＝7.3Hz,1H),7.13(s,1H),5.20(s,2H),3.71(s,3H),3.44(s,2H),2.12(s,6H)。

Step E3- [ (dimethylamino) methyl group]-5-fluoro-1-methyl-1H-indol-6-ol

10% Pd/C (50mg,0.05eq) was added to the flask, followed by vacuum and flushing with nitrogen (× 2). A solution of the product of step D (285mg,0.91mmol,1eq) in ethanol (20mL) was added, the flask was evacuated and flushed with nitrogen (× 3), then evacuated and flushed with hydrogen (× 3), then hydrogenated and shaken at room temperature for 4 h. The reaction was filtered through a short column of HM-N, eluted with ethanol and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C1850 g Gold redissep column) eluting with a gradient of 10-100% acetonitrile in water afforded the desired product as an off white solid (69.8mg,0.27mmol, 29%) (hydrochloride salt).

LC/MS(C₁₂H₁₅FN₂O)178[M+H-NHMe₂]⁺；RT 0.36(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ10.08(s,1H),9.66(s,1H),7.60(d,J＝11.8Hz,1H),7.40(s,1H),6.96(d,J＝7.5Hz,1H),4.29(s,2H),3.71(s,3H),2.66(s,6H),1.23(d,J＝6.5Hz,1H)。

Preparation example 4i4- [4- (dimethylamino) butyl]-2-fluorophenol

Step A[3- (1, 3-dioxo-2, 3-dihydro-1H-isoindol-2-yl) propyl ] ester]Triphenylphosphonium bromide

N- (3-bromopropyl) phthalimide (2.75g,10.26mmol,1eq) and triphenylphosphine (2.69g,10.3mmol,1eq) were stirred in toluene (25mL) and heated at reflux overnight. The reaction was cooled to room temperature, the solid collected by filtration and dried in vacuo to afford the desired product as a white solid (2.55g,4.81mmol, 47%).

¹H NMR(400MHz,DMSO-d6)δ7.95-7.85(m,7H),7.82-7.71(m,12H),3.80-3.64(m,4H),1.99-1.90(m,2H)。

Step B 2- [ (3E) -4- (3-fluoro-4-methoxyphenyl) but-3-en-1-yl]-2, 3-dihydro-1H-isoindole-1, 3-dione

To a solution of the product of step A (2.55g,4.81mmol,1eq) in toluene (25mL) was added 3-fluoro-4-methoxybenzaldehyde (741mg,4.81mmol,1eq) followed by 18-crown-6 (108. mu.L, 0.48mmol,0.1eq) and the mixture was stirred at room temperature overnight. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to afford the desired product as a white solid (1.48g,4.55mmol, 95%).

LC/MS(C₁₉H₁₆FNO₃)302[OTHER]；RT 2.25(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.91-7.79(m,4H),7.13-6.98(m,3H),6.43-6.33(m,1H),5.61(dt,J＝11.7,7.4Hz,1H),3.82(s,3H),3.70(t,2H),2.64(qd,J＝7.2,1.8Hz,2H)。

Step C(3E) -4- (3-fluoro-4-methoxyphenyl) but-3-en-1-amine

To a solution of the product of step B (1.48g,4.55mmol,1eq) in ethanol (60mL) was added methylamine (2M in methanol; 24mL,665mmol,146eq) and the mixture was heated at reflux overnight. The reaction was cooled to room temperature and concentrated in vacuo. The residue was triturated with ether, filtered and dried in vacuo. The crude solid was dissolved in ethyl acetate and extracted with 1N aqueous hydrochloric acid (3 × 100 mL). The combined aqueous extracts were basified with 4M aqueous potassium hydroxide, extracted with ethyl acetate (× 2), the combined organic extracts dried (magnesium sulfate) and concentrated in vacuo to give the desired product as a pink gum (395mg,2.02mmol, 45%).

LC/MS(C₁₁H₁₄FNO)196[M+H]⁺；RT 1.25(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.24-7.04(m,4H),6.44-6.30(m,1H),5.63(dt,J＝11.6,7.2Hz,1H),2.65(t,2H),2.37(qd,J＝7.1,2.0Hz,2H)。

Step D4- (3-fluoro-4-methoxyphenyl) butan-1-amine

To a solution of the product of step C (395mg,2.02mmol,1eq) in methanol (10mL) under nitrogen was added platinum (IV) oxide (45.9mg,0.2mmol,0.1 eq). The vessel was evacuated and backfilled with nitrogen (x3), evacuated, then placed under a hydrogen atmosphere and shaken at room temperature overnight. The reaction was filtered through celite, eluted with methanol and concentrated in vacuo. The residue was dissolved in methanol and loaded onto an SCX cartridge (5g) wetted with methanol, washed with methanol, eluted with 1.75N methanolic ammonia and concentrated in vacuo to afford the desired product as a pink gum (219mg,1.11mmol, 55%).

LC/MS(C₁₁H₁₆FNO)198[M+H]⁺；RT 1.18(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.10-7.01(m,2H),6.99-6.90(m,1H),3.80(s,3H),2.57-2.44(m,2H),1.61-1.42(m,4H),1.39-1.26(m,2H)。

Step E[4- (3-fluoro-4-methoxyphenyl) butyl group]Dimethylamine

To a solution of the product of step D (219mg,1.11mmol,1eq) in methanol (5mL) was added aqueous formaldehyde (37 wt%; 91.8. mu.L, 13.4M,3.33mmol,3eq), sodium triacetoxyborohydride (706mg,3.33mmol,3eq), and glacial acetic acid (6.36. mu.L, 0.11mmol,0.1eq) and the mixture was stirred at room temperature overnight. The reaction was concentrated in vacuo, then partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMSilica gel cartridge) purePurification, eluting with a gradient of 0-10% methanol in dichloromethane, gave the desired product as a clear oil (163mg,0.72mmol, 65%).

LC/MS(C₁₃H₂₀FNO)226[M+H]⁺；RT 1.30(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.10-7.00(m,2H),6.98-6.90(m,1H),3.80(s,3H),2.56-2.47(m,2H),2.23-2.15(m,2H),2.09(s,6H),1.59-1.47(m,2H),1.43-1.31(m,2H)。

Step F4- [4- (dimethylamino) butyl]-2-fluorophenol

To a solution of the product of step E (163mg,0.72mmol,1eq) in dichloromethane (5mL) cooled to 0 deg.C was added boron tribromide (1M in dichloromethane; 2.17mL,2.17mmol,3eq) and the mixture was stirred at room temperature overnight. The reaction was cooled to 0 ℃, quenched with methanol and concentrated in vacuo. The residue was dissolved in methanol and loaded onto an SCX cartridge (5g) wetted with methanol, washed with methanol, eluted with 1.75N methanolic ammonia and concentrated in vacuo to afford the desired product as a yellow oil (110mg,0.52mmol, 72%).

LC/MS(C₁₂H₁₈FNO)212[M+H]⁺；RT 0.96(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ9.54(s,1H),6.94(dd,J＝12.5,2.0Hz,1H),6.88-6.75(m,2H),2.47(t,J＝7.6Hz,2H),2.21-2.13(m,2H),2.08(s,6H),1.56-1.44(m,2H),1.42-1.30(m,2H)。

Preparation example 4jN- [2- (3-fluoro-4-hydroxyphenyl) ethyl]-N-methylcarbamic acid tert-butyl ester

Step AN- [2- (3-fluoro-4-methoxyphenyl) ethyl]Urethane composition

To a solution of 2- (3-fluoro-4-methoxyphenyl) ethan-1-amine (263mg,1.55mmol,1eq) in dichloromethane (10mL) was added triethylamine (315mg,3.11mmol,2eq) and the mixture was cooled to 0 ℃, then ethyl chloroformate (149 μ L,1.55mmol,1eq) was added and the mixture was stirred at room temperature overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel tubePurification, eluting with a gradient of 0-1% methanol in dichloromethane, gave the desired product as a white wax (245mg,1.02mmol, 65%).

LC/MS(C₁₂H₁₆FNO₃)242[M+H]⁺；RT 1.81(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.13(t,1H),7.11-7.01(m,2H),6.98-6.92(m,1H),3.96(q,J＝7.1Hz,2H),3.80(s,3H),3.16(td,J＝7.3,5.8Hz,2H),2.64(t,J＝7.3Hz,2H),1.13(t,J＝7.1Hz,3H)。

Step B[2- (3-fluoro-4-methoxyphenyl) ethyl group](methyl) amine

A solution of the product of step A (245mg,1.02mmol,1eq) in tetrahydrofuran (3mL) was cooled to 0 ℃. Lithium aluminum hydride (1M in tetrahydrofuran, 2.54mL,2.54mmol,2.5eq) was added and the mixture was heated to reflux overnight. The reaction was cooled to 0 ℃ and water (96. mu.L) was added, followed by 15% aqueous sodium hydroxide (96. mu.L) and then water (288. mu.L). Tetrahydrofuran was added to aid stirring and the mixture was stirred at room temperature for 30 minutes. Magnesium sulfate was added, then ethyl acetate was added, and the mixture was stirred for 15 minutes, then filtered through celite and eluted with ethyl acetate. The solvent was removed in vacuo and purified by automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a clear oil (89mg,0.49mmol, 48%).

LC/MS(C₁₀H₁₄FNO)184[M+H]⁺；RT 0.75(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.15-7.01(m,2H),6.96(ddd,J＝8.3,2.0,1.0Hz,1H),3.80(s,3H),2.69-2.57(m,4H),2.27(s,3H)。

Step C2-fluoro-4- [2- (methylamino) ethyl]Phenol and its preparation

To a solution of the product of step B (89mg,0.49mmol,1eq) in dichloromethane (4mL) cooled to 0 ℃ was added boron tribromide (1M in dichloromethane, 1.46mL,1.46mmol,3eq) and the mixture was stirred at room temperature for 4 hours. The reaction was cooled to 0 ℃ and quenched with methanol, then concentrated in vacuo. The residue was dissolved in methanol and loaded onto an SCX cartridge (5g) wetted with methanol, washed with methanol, eluted with 1.75N methanolic ammonia and concentrated in vacuo to afford the desired product as a brown gum (62mg,0.37mmol, 75%).

LC/MS(C₉H₁₂FNO)170[M+H]⁺；RT 0.24(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.02-6.93(m,1H),6.88-6.76(m,2H),2.67-2.53(m,4H),2.27(s,3H)。

Step DN- [2- (3-fluoro-4-hydroxyphenyl) ethyl]-N-methylcarbamic acid tert-butyl ester

To a solution of the product of step C (62mg,0.37mmol,1eq) in dichloromethane (5mL) was added triethylamine (153. mu.L, 1.1mmol,3eq) and 4- (dimethylamino) pyridine (4.48mg,0.04mmol,0.1eq), followed by di-tert-butyl dicarbonate (0.09mL,0.44mmol,1.2eq) and the mixture was stirred at room temperature for 3 hours. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-85% ethyl acetate in isoheptane to give the desired product as a clear oil (48mg,0.18mmol, 49%).

LC/MS(C₁₄H₂₀FNO₃)170[M-Boc+H]⁺；RT 2.54(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ9.60(s,1H),6.95(d,J＝12.3Hz,1H),6.89-6.72(m,2H),3.34-3.27(m,2H),2.73(s,3H),2.64(t,J＝7.1Hz,2H),1.28(s,9H)。

Preparation example 4kN- [4- (3-fluoro-4-hydroxyphenyl) butyl]-N-methylcarbamic acid tert-butyl ester

Step AN- [ (3E) -4- (3-fluoro-4-methoxyphenyl) but-3-en-1-yl]Urethane composition

To a solution of the product of preparation 4i, step C (397mg,2.03mmol,1eq) in dichloromethane (20mL) was added triethylamine (0.57mL,4.07mmol,2eq) and the mixture was cooled to 0 ℃. Ethyl chloroformate (194. mu.L, 2.03mmol,1eq) was added and the mixture was warmed to room temperature and stirred overnight. The reaction solution was partitioned between dichloromethane and saturated aqueous sodium bicarbonate solution, and the organic phase was separated Separate (PTFE phase separator) and concentrate in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-25% ethyl acetate in isoheptane to give the desired product as a clear oil (310mg,1.16mmol, 57%).

LC/MS(C₁₄H₁₈FNO₃)268[M+H]⁺；RT 2.06(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.24-7.05(m,3H),6.42-6.33(m,1H),5.57(dt,J＝11.7,7.2Hz,1H),4.00(dq,J＝26.6,7.1Hz,2H),3.84(s,3H),3.08(q,J＝6.8Hz,2H),2.42(qd,J＝7.1,1.9Hz,2H),1.16(t,3H)。

Step BN- [4- (3-fluoro-4-methoxyphenyl) butyl]Urethane composition

To a solution of the product of step A (310mg,1.16mmol,1eq) in methanol (12mL) under nitrogen was added platinum (IV) oxide (26.3mg,0.12mmol,0.1 eq). The vessel was evacuated and backfilled with nitrogen (x3), evacuated, placed under a hydrogen atmosphere and shaken at room temperature overnight. The reaction was filtered through celite, eluted with methanol and concentrated in vacuo to afford the desired product as a clear oil (275mg,1.02mmol, 88%).

LC/MS(C₁₄H₂₀FNO₃)270[M+H]⁺；RT 2.07(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.12-7.00(m,3H),6.99-6.91(m,1H),3.96(q,J＝7.1Hz,2H),3.80(s,3H),2.97(q,J＝6.7Hz,2H),2.52-2.45(m,2H),1.51(p,J＝7.8,7.4Hz,2H),1.37(p,J＝7.2Hz,2H),1.14(t,J＝7.1Hz,3H)。

Step C[4- (3-fluoro-4-methoxyphenyl) butyl group](methyl) amine

To a solution of the product of step B (417mg,1.55mmol,1eq) in tetrahydrofuran (5mL) cooled to 0 deg.C was added lithium aluminum hydride (1M in tetrahydrofuran; 3.87mL,3.87mmol,2.5eq) and the mixture was heated at reflux overnight. The reaction was cooled to 0 ℃ and water (150. mu.L) was added, followed by 15% aqueous sodium hydroxide (150. mu.L) and water (450. mu.L). The mixture was diluted with tetrahydrofuran and stirred for 30 minutes. Magnesium sulfate and ethyl acetate were added and the mixture was filtered through celite and concentrated in vacuo. By automation Flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-15% methanol in dichloromethane afforded the desired product as a clear oil (222mg,1.05mmol, 68%).

LC/MS(C₁₂H₁₈FNO)212[M+H]⁺；RT 1.28(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.09-6.99(m,2H),6.94(dd,1H),3.80(s,3H),2.51-2.48(m,2H),2.44(t,2H),2.24(s,3H),1.61-1.47(m,2H),1.47-1.31(m,2H)。

Step D2-fluoro-4- [4- (methylamino) butyl]Phenol and its preparation

To a solution of the product of step C (222mg,1.05mmol,1eq) in dichloromethane (10mL) cooled to 0 ℃ was added boron tribromide (1M in dichloromethane, 3.15mL,3.15mmol,3eq) and the mixture was stirred at room temperature for 3 hours. The reaction was cooled to 0 ℃, quenched with methanol and concentrated in vacuo. The residue was dissolved in methanol and loaded onto an SCX cartridge (5g) wetted with methanol, washed with methanol, eluted with 1.4N methanolic ammonia and concentrated in vacuo to afford the desired product as a brown gum (63mg,0.32mmol, 30%).

LC/MS(C₁₁H₁₆FNO)198[M+H]⁺；RT 1.01(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.00-6.91(m,1H),6.88-6.75(m,2H),2.49-2.40(m,4H),2.24(s,3H),1.58-1.45(m,2H),1.43-1.33(m,2H)。

Step EN- [4- (3-fluoro-4-hydroxyphenyl) butyl]-N-methylcarbamic acid tert-butyl ester

To a solution of the product of step D (63mg,0.32mmol,1eq) in dichloromethane (5mL) was added triethylamine (133. mu.L, 0.96mmol,3eq) and 4- (dimethylamino) pyridine (3.9mg,0.03mmol,0.1eq), the mixture was cooled to 0 ℃ and di-tert-butyl dicarbonate (66. mu.L, 0.29mmol,0.9eq) was added and the mixture was stirred at room temperature for 1 hour. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMSilica gel cartridge), gradient eluting with 0-35% ethyl acetate in isoheptane to obtain the desired productProduct (33mg,0.11mmol, 35%).

LC/MS(C₁₆H₂₄FNO₃)198[M-Boc+H]⁺；RT 2.18(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ9.54(s,1H),6.95(dd,J＝12.4,2.0Hz,1H),6.88-6.74(m,2H),3.21-3.11(m,2H),2.74(s,3H),2.49-2.41(m,2H),1.53-1.40(m,4H),1.37(s,9H)。

Preparation example 5a1- (1-adamantylmethyl) -5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazole

Step A1- (1-adamantylmethyl) -4-iodo-pyrazoles

A mixture of 35.9g of 1-adamantylmethanol (216mmol), 73.48g of triphenylphosphine (280mmol,1.3eq.), 54.25g of 4-iodo-1H-pyrazole (280mmol,1.3eq.) and 64.4g N- (tert-butoxycarbonyliminomethylene) carbamic acid tert-butyl ester (266mmol.1.3eq.) in 1078mL of THF was stirred at room temperature for 48H. After addition of 10.94g of 4-iodo-1H-pyrazole (56mmol,0.26eq.), 12.81g N tert-butyl (tert-butoxycarbonyliminomethylene) carbamate (53mmol,0.26eq.) and 14.69g of triphenylphosphine (56mmol,0.26eq.), the reaction mixture was stirred at room temperature for 24H, then concentrated, purified by flash column chromatography using DCM as eluent, triturated with cold MeOH and filtered off to give 53.6g (73%) of the desired product as a white powder.

Step B1- (1-adamantylmethyl) -4-iodo-5-methyl-pyrazole

To a solution of 9.8mL of diisopropylamine (69.5mmol,1.1eq.) in 180mL of THF at-78 ℃ was added dropwise a solution of 33.4mL of 2.5m butyllithium (84mmol,1.3eq.) and the mixture was stirred at-78 ℃ for 0.5 h, treated with 22.0g of the product of step a (64.28mmol,1eq.) in 90mL of THF, stirred at-78 ℃ for 1h, treated with 4.67mL of iodomethane (73.3mmol,1.14eq.) and stirred at-78 ℃ for 18 h. With concentrated NH ₄The reaction was quenched with EtOAc and the combined organic phases were washed with brine, dried, concentrated, triturated with MeOH and filtered to give 21g (92%) of the desired product.

Step C1- (1-adamantylmethyl) -5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazole

To a solution of 21g of the product of step B (58.95mmol,1eq.) in 300mL THF at-78 deg.C was added 28.3mL of a 2.5M solution of butyllithium (70.8mmol,1.2eq) and the mixture was stirred at-78 deg.C for 0.5 h, treated with 16.4g of 2-isopropoxy-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan (88.1mmol,1.5eq.) added in portions over 40 min and held at-78 deg.C for 24 h. With concentrated NH at room temperature₄The reaction was quenched with EtOAc and the combined organic phases were washed with brine, dried, concentrated, triturated with MeOH and filtered to give 19.7g (94%) of the desired product as off-white crystals.

¹H NMR(500MHz,DMSO-d₆)δppm 7.45(s,1H),3.69(s,2H),2.36(s,3H),1.91(m,1H),1.64/1.54(m,6H),1.50(m,6H),1.24(s,12H)；¹³C NMR(500MHz,,DMSO-d₆)δppm 146.9,144.1,104.6,59.7,40.6,36.8,35.4,28.1,25.1,12.1；HRMS-ESI(m/z):[M+H]+C₂₁H₃₄BN₂O₂357.2713, found 357.2704.

Preparation example 5b1- { [1- (3-methoxypropyl) cyclooctyl radical]Methyl } -5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole

Step A1- (3-methoxypropyl) cyclooctanecarboxylic acid methyl ester

To a solution of 4.74g (1.14eq.) of diisopropylamine in 90mL of tetrahydrofuran was added 18.8mL (1.14eq.) of a 2.5M solution of butyllithium at-78 ℃ and after holding at-78 ℃ for 0.5 hour, a solution of 7.0g (41.1mmol) of methyl cyclooctanecarboxylate in 40mL of tetrahydrofuran was added over 1 hour. After 1 hour at-78 ℃ 7.2g (1.14eq.) of 1-bromo-3-methoxy-propane were added and the mixture was stirred for 18 hours. Adding saturated NH ₄After the reaction was quenched with Cl solution, the mixture was extracted with EtOAc and the organic phase was MgSO₄Drying and concentration gave 8.0g (80%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 3.66(s,3H),3.33(t,2H),3.31(s,3H),2.03-1.94(m,2H),1.64-1.38(m,16H)。

Step B1- (3-methoxypropyl) cyclooctyl]Methanol

To a solution of 9.0g (37.13mmol) of the product of step A in 93mL of diethyl ether at 0 ℃ 1.76g (1.25eq.) of lithium aluminium hydride are added in portions. After stirring at room temperature for 2 hours, the reaction was quenched by the addition of ice water and EtOAc and 10% NaOH solution were added. The mixture was extracted with EtOAc, dried and concentrated to give 7.4g (93%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 3.37(t,2H),3.34(s,3H),3.30(s,2H),1.61-1.23(m,18H)。

Step C4-iodo-1- [ [1- (3-methoxypropyl) cyclooctyl group]Methyl radical]-1H-pyrazoles

To a solution of 1.39g (6.5mmol) of the product of step B and 1.64g (1.3eq.) of 4-iodo-1H-pyrazole in 33mL of tetrahydrofuran were added 2.22g (1.3eq.) of triphenylphosphine and 1.95g (1.3eq.) of di-tert-butyl azodicarboxylate and the mixture was stirred at room temperature for 67 hours. 278mg of 4-iodo-1H-pyrazole, 444mg of triphenylphosphine and 390mg of di-tert-butyl azodicarboxylate were added to the mixture and stirred at room temperature for 24 hours. The addition of the reagent was repeated and stirred at room temperature for 24 hours (115 hours total stirring), then the mixture was concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluent to give 1.24g (49%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 7.47(s,1H),7.42(s,1H),3.93(s,2H),3.37(t,2H),3.36(s,3H),1.68-1.18(m,18H)。

Step D4-iodo-1- [ [1- (3-methoxypropyl) cyclooctyl group]Methyl radical]-5-methyl-1H-pyrazoles

To a solution of 1.2g (3.07mmol) of the product of step C in 5mL of tetrahydrofuran was added 3.7mL (1.2eq.) of a 1M LDA solution at-78 ℃. After holding at-78 ℃ for 0.6 h, 0.5mL (1.14eq.) of iodomethane was added dropwise to the mixture and the temperature was raised to room temperature over 20 h. With saturated NH₄The reaction was quenched with Cl solution and extracted with EtOAc. The combined organic phases were dried, concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluent to give 0.79g (64%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 7.43(s,1H),3.85(s,2H),3.38(t,2H),3.35(s,3H),2.29(s,3H),1.69-1.24(m,18H)。

Step E1- [ [1- (3-methoxypropyl) cyclooctyl group]Methyl radical]-5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole

To a solution of 0.81g (2mmol) of the product of step D in 15mL of tetrahydrofuran is added dropwise a solution of 0.96mL (1.2eq.) of 2.5M butyllithium at-78 ℃. After 0.5 h, 0.5mL (1.2eq.) of 2-isopropoxy-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolane was added over 20 minutes and the mixture was held at-78 ℃ for 6 hours and then at room temperature for 6 hours. With saturated NH₄The reaction was quenched with Cl solution and extracted with EtOAc, and the combined organic phases were washed with brine, dried and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluent to give 0.33g (34%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 7.46(s,1H),3.75(s,2H),3.27(t,2H),3.21(s,3H),2.36(s,3H),1.66-1.1(m,14H),1.57(m,2H),1.24(s,12H),1.24(m,2H)。¹³C NMR(500MHz,dmso-d6)δppm 147.3,144.5,104.5,73.2,58.2,54.4,40.5,33.2,25.1,23.6,11.8.IR:2922,1556,1246,1144,1055.HRMS-ESI(m/z):[M+H]+C₂₃H₄₂N₂O₃Calculated value of B405.3289, found value 405.3329.

Preparation example 5c1- { [1- (3-methoxypropyl) cyclohexyl]Methyl } -5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole

Step A1- (3-methoxypropyl) cyclohexanecarboxylic acid methyl ester

To a solution of 6.84g (1.09eq.) of diisopropylamine in 130mL of tetrahydrofuran was added 27mL (1.09eq.) of a 2.5M solution of butyllithium at-78 ℃ and held at-78 ℃ for 0.5 hour, and over 1 hour a solution of 8.8g of methyl cyclohexanecarboxylate in 50mL of tetrahydrofuran. After 1 hour at-78 ℃, 10.7g (1.13eq.) of 1-bromo-3-methoxy-propane was added and the mixture was stirred for 18 hours. By adding saturated NH₄The reaction was quenched with Cl solution, the mixture was extracted with EtOAc and the organic phase was MgSO₄Drying and concentration gave 12g (92%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 3.67(s,3H),3.35(d,1H),3.32(d,1H),3.31(s,3H),2.11-2.03(m,2H),1.60-1.16(m,12H)。

Step B1- (3-methoxypropyl) cyclohexyl]Methanol

To a solution of 12g (56.41mmol) of the product of step A in 140mL of diethyl ether at 0 ℃ 2.68g (1.25eq.) of lithium aluminium hydride are added in portions. After stirring at room temperature for 2 hours, the reaction was quenched by the addition of ice water and EtOAc and 10% NaOH solution were added. The mixture was extracted with EtOAc, dried and concentrated to give 9.37g (89%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 3.41(s,2H),3.38(t,2H),3.35(s,3H),1.56-1.27(m,14H)。

Step C4-iodo-1- [ [1- (3-methoxypropyl) cyclohexyl ] carbonyl ]Methyl radical]Pyrazoles

To a solution of 1.21g (6.5mmol) of the product of step B and 2.58g (2.05eq.) of 4-iodo-1H-pyrazole in 33mL of tetrahydrofuran were added 3.5g (2.05eq.) of triphenylphosphine and 3.07g (2.05eq.) of di-tert-butyl azodicarboxylate and the mixture was stirred at room temperature for 2 hours. To the mixture were added 140mg of 4-iodo-1H-pyrazole, 230mg of triphenylphosphine and 200mg of di-tert-butyl azodicarboxylate and stirred at room temperature for 24 hours. The steps of adding the reagent and stirring at room temperature for 24 hours were repeated twice (96 hours stirring total), then the mixture was concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluent to give 1.4g (59.5%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 7.47(s,1H),7.41(s,1H),4.00(s,2H),3.36(t,2H),3.35(s,3H),1.62-1.21(m,14H)。

Step D4-iodo-1- [ [1- (3-methoxypropyl) cyclohexyl ] carbonyl]Methyl radical]-5-methyl-pyrazole

To a solution of 3.7g (10.21mmol) of the product of step C in 15mL of tetrahydrofuran was added 12.3mL (1.2eq.) of a 1M solution of LDA in tetrahydrofuran at-78 ℃. After 0.6 h at-78 ℃, 0.73mL (1.14eq.) of methyl iodide was added dropwise to the mixture and warmed to room temperature over 20 h. With saturated NH₄The reaction was quenched with Cl solution and extracted with EtOAc. The combined organic phases were dried, concentrated and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluent to give 2.85g (74%) of the desired product.

¹H NMR(400MHz,CDCl₃)δppm 7.44(s,1H),3.92(s,2H),3.38(t,2H),3.35(s,3H),2.29(s,3H),1.58-1.13(m,14H)。

Step E1- [ [1- (3-methoxypropyl) cyclohexyl ] carbonyl]Methyl radical]-5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazole

To a solution of 5.0g (13.3mmol) of the product of step D in 71mL of tetrahydrofuran was added dropwise 6.38mL (1.2eq.) of a 2.5 molar solution of butyllithium at-78 ℃. After 0.5 hour, 4.1mL (1.5eq.) of 2-isopropoxy-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolane was added over 40 minutes and the mixture was held at-78 ℃ for 6 hours and then at room temperature for 6 hours. With saturated NH₄After the reaction was quenched with Cl solution and extracted with EtOAc, the combined organic phases were washed with brine, dried and purified by flash column chromatography (silica gel) using heptane and EtOAc as eluent to give 2.3g (46%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 7.47(s,1H),3.84(s,2H),3.27(t,2H),3.2(s,3H),2.37(s,3H),1.54-1.07(m,10H),1.46(m,2H),1.32(m,2H),1.24(s,12H)。¹³C NMR(500MHz,dmso-d6)δppm 147.3,144.4,104.6,73.1,58.2,55.7,37.9,30.6,25.1,23.1,12.0.IR:2927,1556,1257,1144,1053.HRMS-ESI(m/z):[M+H]Calculated value C of +₂₁H₃₈N₂O₃376.2897, found 376.3019.

Preparation example 6a5-bromo-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

Step A2- [ (pent-3-yn-1-yl) amino group]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of ethyl 2-bromo-1, 3-thiazole-4-carboxylate (538mg,2.28mmol,1eq) in acetonitrile (10mL) was added pent-3-yn-1-amine hydrochloride (300mg,2.51mmol,1.1eq) and triethylamine (0.7mL,5.02mmol,2.2eq) and the mixture was heated under microwave irradiation at 150 ℃ for 3 hours. The reaction was partitioned between ethyl acetate and brine, the organic phase was dried (magnesium sulfate) and And (4) concentrating in vacuum. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to afford the desired product as a beige solid (221mg,0.93mmol, 41%). LC/MS (C)₁₁H₁₄N₂O₂S)239[M+H]⁺；RT 2.22(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.93(t,J＝5.7Hz,1H),7.52(s,1H),4.22(q,J＝7.1Hz,2H),3.38-3.28(m,2H),2.44-2.33(m,2H),1.75(t,J＝2.5Hz,3H),1.27(t,J＝7.1Hz,3H)。

Step B2- { 3-chloro-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (140mg,0.93mmol,1eq) in tetrahydrofuran was added the product of step A (221mg,0.93mmol,1eq) and the mixture was heated under reflux overnight. The reaction was concentrated in vacuo and the residue triturated with dichloromethane, filtered and dried in vacuo to afford the desired product as an off-white solid (148mg,0.46mmol, 49%).

LC/MS(C₁₃H₁₃ClN₄O₂S)325[M+H]⁺；RT 2.32(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.11(s,1H),4.41(dd,J＝8.8,7.7Hz,2H),4.30(q,J＝7.1Hz,2H),3.34-3.24(m,2H),2.29(d,J＝1.1Hz,3H),1.31(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step B (148mg,0.46mmol,1eq), 2-aminobenzothiazole (103mg,0.68mmol,1.5eq), XantPhos (52.7mg,0.09mmol,0.2eq), cesium carbonate (297mg,0.91mmol,2eq) and 1, 4-dioxane (20mL) and the vessel was evacuated and flushed with nitrogen, then tris (dibenzylideneacetone) dipalladium (0) (41.7mg,0.05mmol,0.1eq) was added and the mixture was purged with nitrogen (10min) and then heated under microwave irradiation at 150 ℃ for 2 hours. The reaction was diluted with ethyl acetate, filtered through celite, washed with brine, dried (magnesium sulfate) and concentrated in vacuo And (4) shrinking. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow solid (103mg,0.23mmol, 52%).

LC/MS(C₂₀H₁₈N₆O₂S₂)439[M+H]⁺；RT 2.67(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ10.94(br s,1H),8.06(s,1H),7.95(br s,1H),7.66(br s,1H),7.44-7.33(m,1H),7.28-7.15(m,1H),4.42-4.34(m,2H),4.30(q,2H),3.32-3.28(m,2H),2.34(s,3H),1.32(t,J＝7.1,2.4Hz,3H)。

Step D2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a cooled solution of the product of step C (103mg,0.23mmol,1eq) in tetrahydrofuran (15mL) and dimethylformamide (5mL) was added N, N-diisopropylethylamine (81.8. mu.L, 0.47mmol,2 eq). After 5 min 4-dimethylaminopyridine (5.74mg,0.05mmol,0.2eq) and [2- (chloromethoxy) ethyl ] were added]Trimethylsilane (103 μ L,0.59mmol,2.5eq) and the mixture was stirred at room temperature overnight. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to afford the desired product as an off white solid (98mg,0.17mmol, 73%).

LC/MS(C₂₆H₃₂N₆O₃SiS₂) Not ionized; RT 3.08(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.14(s,1H),7.91(d,J＝7.6Hz,1H),7.60-7.51(m,2H),7.39-7.30(m,1H),5.96(s,2H),4.48(t,J＝8.1Hz,2H),4.40(q,J＝7.1Hz,2H),3.87-3.78(m,2H),3.48-3.36(m,2H),2.44(s,3H),1.42(t,J＝7.1Hz,3H),1.07-0.98(m,2H),0.00(s,9H)。

Step E 5-bromo-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl radical} -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step D (98mg,0.17mmol,1eq) in dichloromethane (15mL) was added N-bromosuccinimide (39.9mg,0.22mmol,1.3eq) and the mixture was stirred at room temperature for 3 hours. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to afford the desired product as an off white solid (98mg,0.15mmol, 88%).

LC/MS(C₂₆H₃₁BrN₆O₃SiS₂) Not ionized; RT 3.22(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.81(dd,J＝7.6,1.1Hz,1H),7.50-7.39(m,2H),7.28-7.21(m,1H),5.85(s,2H),4.40-4.24(m,4H),3.68-3.58(m,2H),3.27(t,J＝8.0Hz,2H),2.32(s,3H),1.31(t,J＝7.1Hz,3H),1.02(dd,J＝8.5,7.4Hz,2H),-0.12(s,9H)。

Preparation example 7Tert-butyl-diphenyl- [2- [ [3, 5-dimethyl-7- [ [ 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl]Methyl radical]-1-adamantyl]Oxy radical]Ethoxy radical]Silane

Step A3-bromo-5, 7-dimethyladamantane-1-carboxylic acid

Iron (6.7g,120mmol) was stirred in bromine (30.7mL,600mmol,5eq) at 0 ℃ for 1 h, then 3, 5-dimethyladamantane-1-carboxylic acid (25g,1eq) was added and the reaction mixture was stirred at room temperature for 2 days. After addition of EtOAc, the reaction mixture was carefully treated with saturated sodium thiosulfate solution at 0 ℃ and stirred for 15 minutes. After filtration through a pad of celite and washing with EtOAc, the organic phase was separated, washed with saturated sodium thiosulfate solution and brine, dried, and concentrated to give the desired product (34.28g, 74.6%) which was used without further purification.

¹H NMR(400MHz,DMSO-d₆):δppm 12.33(br.,1H),2.21(s,2H),1.96/1.91(d+d,4H),1.50/1.43(d+d,4H),1.21/1.14(dm+dm,2H),0.86(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 176.8,66.8,54.0,48.7,48.5,45.7,43.3,35.5,29.4；HRMS-ESI(m/z):[M-H]-C₁₃H₁₈BrO₂285.0496; found 285.0498.

And B:3-bromo-5, 7-dimethyl-1-adamantyl-methanol

To the product of step A (34.3g,119mmol) in THF (77.6mL) was slowly added 1M BH₃A solution of THF (358mL,3eq) and the reaction mixture stirred for 18 hours. After addition of methanol and stirring for 30 min, purification by column chromatography (silica gel, heptane and MTBE as eluent) gave the desired product (16.19g, 49.6%).

¹H NMR(400MHz,DMSO-d₆):δppm 4.51(t,1H),3.05(d,2H),1.91(s,2H),1.91(s,4H),1.19/1.09(d+d,2H),1.19/1.05(d+d,4H),0.85(s,6H)¹³C NMR(100MHz,DMSO-d₆)δppm70.4,68.9,54.9,49.8,49.3,43.8,41.4,35.7,29.7；HRMS-ESI(m/z):[M-Br]-C₁₃H₂₁Calculated value of O193.1598; found 193.1589.

Step C1- [ 3-bromo-5, 7-dimethyl-1-adamantyl]Methyl radical]Pyrazoles

To a solution of the product of step B (16.19g,59.26mmol) and 1H-pyrazole (4.841g,1.2eq) in toluene (178mL) was added cyanomethylenetributylphosphine (18.64mL,1.2eq) in one portion and the reaction mixture was stirred at 90 ℃ for 2H. Purification by column chromatography (silica gel, heptane and MTBE as eluent) gave the desired product (17.88g, 93%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.63(d,1H),7.43(d,1H),6.23(t,1H),3.90(s,2H),1.92-1.02(m,12H),0.83(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.0,131.8,105.2,67.7,61.4,54.4/48.8/44.6,50.4,35.7,29.6；HRMS-ESI(m/z):[M]+C₁₆H₂₃BrN₂322.1045 found value of 322.1014.

Step D:5-methyl-1- [ [ -3-bromo-5, 7-dimethyl-1-adamantyl]Methyl radical]Pyrazoles

To the product of step C (17.88g,55.3mmol) in THF (277mL) was added butyllithium (2.5M in THF, 66mL,3eq) at-78 deg.C and after 1 hour methyl iodide (17.2mL,5 eq). After 10 minutes, the reaction was mixed Saturated NH for compounds₄The Cl solution was quenched, extracted with EtOAc and the combined organic layers were dried and concentrated to give the desired product (18.7g, 100%) which was used in the next step without further purification.

¹H NMR(400MHz,DMSO-d₆):δppm 7.31(d,1H),6.00(d,1H),3.79(s,2H),2.23(s,3H),2.01(s,2H),1.89/1.85(d+d,4H),1.23/1.15(d+d,4H),1.16/1.05(d+d,2H),0.83(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.2,138.0,105.2,67.8,57.8,54.4,50.6,48.8,44.8,41.5,35.7,29.6,11.8；HRMS-ESI(m/z):[M+H]+C₁₇H₂₆BrN₂337.1279 found value of 337.1289.

Step E:2- [ [ -3, 5-dimethyl-7- [ (5-methylpyrazol-1-yl) methyl ] methyl]-1-adamantyl]Oxy radical]Ethanol

A mixture of the product of step D (18.7g,55.3mmol), ethylene glycol (123mL,40eq) and DIPEA (48.2mL,5eq) was stirred at 120 ℃ for 6 h. After the reaction mixture was diluted with water and extracted with EtOAc, the combined organic layers were dried and concentrated to give the desired product (18.5g, 105%) which was used in the next step without further purification.

¹H NMR(400MHz,DMSO-d₆):δppm 7.29(d,1H),5.99(d,1H),4.45(t,1H),3.78(s,2H),3.39(q,2H),3.32(t,2H),2.23(s,3H),1.34(s,2H),1.27/1.21(d+d,4H),1.13/1.07(d+d,4H),1.04/0.97(d+d,2H),0.84(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.0,137.8,105.1,74.0,62.1,61.5,58.5,50.1,47.0,46.1,43.3,39.7,33.5,30.2,11.9；HRMS-ESI(m/z):[M+H]+C₁₉H₃₁N₂O₂319.2386 found value of 319.2387.

Step FTert-butyl-diphenyl- [2- [ [ -3, 5-dimethyl-7- [ (5-methylpyrazol-1-yl) methyl ] methyl]-1-adamantyl]Oxy radical]Ethoxy radical]Silane

To a mixture of the product of step E (17.6g,55.3mmol) and imidazole (5.65g,1.5eq) in DCM (150ml) was added tert-butyl-chloro-diphenyl-silane (18.6g,1.2eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, heptane and MTBE as eluent) gave the desired product (27.0g, 87.8%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.72-7.34(m,10H),7.29(d,1H),5.99(br.,1H),3.78(s,2H),3.67(t,2H),3.44(t,2H),2.21(s,3H),1.33(s,2H),1.26/1.18(d+d,4H),1.12/1.06(d+d,4H),1.03/0.96(d+d,2H),0.98(s,9H),0.82(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.0,137.8,105.1,74.2,64.4,61.7,58.5,50.0,46.9,46.0,43.4,39.6,33.5,30.1,27.1,19.3,11.9；HRMS-ESI(m/z):[M+H]+C₃₅H₄₉N₂O₂Calculated value of Si 557.3563 found value is 557.3564.

G:tert-butyl-diphenyl- [2- [ [3- [ (4-iodo-5-methyl-pyrazol-1-yl) methyl ] methyl]-5, 7-dimethyl-1-adamantyl]Oxy radical]Ethoxy radical]Silane

To the product of step F (27.0g,48.56mmol) in DMF (243mL) was added N-iodosuccinimide (13.6g,1.25eq) and the reaction mixture was stirred for 2 h. After dilution with water, the mixture was extracted with DCM. The combined organic layers were washed with saturated sodium thiosulfate solution and brine, dried and concentrated to give the desired product (30.1g, 90%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.68-7.37(m,10H),7.45(s,1H),3.89(s,2H),3.67(t,2H),3.44(t,2H),2.23(s,3H),1.30(s,2H),1.26/1.17(d+d,4H),1.12/1.05(d+d,4H),1.00/0.96(d+d,2H),0.98(s,9H),0.82(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 142.5,140.8,133.7,64.4,61.7,60.3,59.9,49.9,46.8,45.9,43.2,39.7,33.5,30.1,27.1,19.3,12.2；HRMS-ESI(m/z):[M+H]+C₃₅H₄₈IN₂O₂Calculated value of Si 683.2530 found value is 683.2533.

Step H:tert-butyl-diphenyl- [2- [ [3, 5-dimethyl-7- [ [ 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl]Methyl radical]-1-adamantyl]Oxy radical]Ethoxy radical]Silane

To the product of step G (17.5G,25.6mmol) in THF (128mL) was added chloro (isopropyl) magnesium-LiCl (1.3M in THF, 24mL,1.2eq) at 0 deg.C, stirred for 40 min, treated with 2-isopropoxy-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (15.7mL,3eq) and the reaction was runThe mixture was stirred for 10 minutes. With saturated NH₄After dilution with Cl solution and extraction with EtOAc, the combined organic phases were concentrated and purified by column chromatography (silica gel, heptane and MTBE as eluent) to give the desired product (15.2g, 86.9%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.65(dm,4H),7.47(s,1H),7.45(tm,2H),7.40(tm,4H),3.80(s,2H),3.66(t,2H),3.44(t,2H),2.35(s,3H),1.35-0.94(m,12H),1.24(s,12H),0.97(s,9H),0.83(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 146.9,144.3,135.6,130.2,128.2,104.7,83.0,74.2,64.4,61.7,58.4,30.1,27.1,25.2,19.3,12.0；HRMS-ESI(m/z):[M+H]+C₄₁H₆₀BN₂O₄Calculated Si 683.4415 found 683.4423.

Preparation example 8:tert-butyl- [3- [3, 5-dimethyl-7- [ [ 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl]Methyl radical]-1-adamantyl]Propoxy group]-diphenyl-silane

Step A1- [ [ 3-allyl-5, 7-dimethyl-1-adamantyl group]Methyl radical]-5-methyl-pyrazole

To the product of preparation 7, step D (15.66g,46.43mmol) and AgOTf (597Mg,0.05eq) in THF (232mL) was added a 2M solution of allyl-Mg-Cl in THF (46.4mL,2eq) and the reaction mixture was stirred for 0.5 h. With saturated NH₄After the reaction was quenched with Cl solution and extracted with EtOAc, the combined organic phases were concentrated and purified by column chromatography (silica gel, heptane and MTBE as eluent) to give the desired product (11.32g, 81.7%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.27(d,1H),5.98(m,1H),5.76(m,1H),5.01/4.96(dm+dm,2H),3.73(s,2H),2.22(s,3H),1.83(d,2H),1.15-0.93(m,12H),0.78(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.0,137.7,135.0,117.7,105.0,59.0,47.8,44.2,35.0,31.8,30.6,11.9；HRMS-ESI(m/z):[M+H]+C₂₀H₃₁N₂299.2487 found value of 299.2485.

And B:3- [3, 5-dimethyl-7- [ (5-methylpyrazol-1-yl) methyl group]-1-adamantyl]Propan-1-ol

To a solution of the product of step A (10.2g,34.17mmol) in THF (85mL) was added 1M BH₃THF solution of THF (85.4mL,2eq) and the reaction mixture was stirred for 1 hour. After treatment with 10M NaOH solution (24mL,7eq) and 33% hydrogen peroxide solution (73mL,25eq) at 0 ℃ the reaction mixture was stirred at room temperature for 1 hour. The reaction was then quenched with aqueous HCl, extracted with EtOAc and purified by column chromatography (silica gel, heptane and MTBE as eluent) to give the desired product (9.75g, 90%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.28(d,1H),5.98(m,1H),4.33(t,1H),3.73(s,2H),3.32(m,2H),2.22(brs,3H),1.32(m,2H),1.12-0.92(m,12H),1.06(m,2H),0.78(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 137.7,105.0,62.1,59.1,39.7,30.7,26.5,11.9,HRMS-ESI(m/z):[M+H]+C₂₀H₃₃N₂Calculated value of O317.2593 found value of 317.2590

And C:tert-butyl- [3- [3, 5-dimethyl-7- [ (5-methylpyrazol-1-yl) methyl group]-1-adamantyl]Propoxy group]-diphenyl-silane

To a solution of the product of step B (9.75g,30.8mmol) and imidazole (3.1g,1.5eq) in DCM (92mL) was added tert-butyl-chloro-diphenyl-silane (9.45mL,1.2eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, heptane and MTBE as eluent) gave the desired product (12.5g, 73%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.63-7.39(m,10H),7.27(d,1H),5.98(d,1H),3.72(s,2H),3.59(t,2H),2.21(s,3H),1.42(m,2H),1.1-0.92(br.,12H),1.09(m,2H),0.98(s,9H),0.77(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 137.7,105.0,64.8,59.1,39.3,38.0,34.2,31.8,30.6,27.2,26.1,19.2,11.9；HRMS-ESI(m/z):[M+H]+C₃₆H₅₁N₂Calculated OSi 555.3771 found 555.3770.

Step DTert-butyl- [3- [3- [ (4-iodo-5-methyl-pyrazol-1-yl) methyl group]-5, 7-dimethyl-1-adamantyl]Propoxy group]-diphenyl-silane

To a solution of the product of step C (12.5g,22.54mmol) in DMF (112mL) was added N-iodosuccinimide (6.34g,1.25eq) and the reaction mixture was stirred for 2 h. After quenching the reaction with saturated sodium thiosulfate solution and extraction with DCM, the combined organic phases were washed with saturated sodium thiosulfate and brine, dried and evaporated to give the desired product (16.3g, 105%).

LC/MS(C₃₆H₅₀IN₂OSi)681[M+H]⁺。

Step E:tert-butyl- [3- [3, 5-dimethyl-7- [ [ 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl]Methyl radical]-1-adamantyl ]Propoxy group]-diphenyl-silane

To the product of step D (16.25g,23.9mmol) in THF (119mL) was added chloro (isopropyl) magnesium-LiCl (1.3M in THF, 22mL,1.2eq.) at 0 deg.C, the mixture was stirred for 40 minutes, treated with 2-isopropoxy-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (14.6mL,3 eq.) and stirred for 10 minutes. With saturated NH₄After dilution with Cl solution and extraction with EtOAc, the combined organic phases were concentrated and purified by column chromatography (silica gel, heptane and MTBE as eluent) to give the desired product (11.4g, 70%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.59(d,4H),7.46(s,1H),7.45(t,2H),7.43(t,4H),3.74(s,2H),3.59(t,2H),2.35(s,3H),1.41(qn,2H),1.24(s,12H),1.09(m,2H),1.08(s,4H),1.05(s,2H),0.98(s,9H),0.98(s,2H),0.94(s,4H),0.78(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 146.9,144.2,135.5,133.8,130.3,128.3,104.6,83.0,64.7,64.7,59.0,50.6,48.2,46.5,44.1,39.2,37.9,31.8,30.7,27.2,26.1,25.2,19.2,12.0；HRMS-ESI(m/z):[M+H]⁺C₄₂H₆₂BN₂O₃Calculated value of Si 681.4623 found value is 681.4631.

Preparation example 9:tert-butyl- [2- [ [3- [ [ 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl]Methyl radical]-1-adamantyl]Oxy radical]Ethoxy radical]-diphenyl-silane

Step A(3-bromo-1-adamantyl) methanol

To a solution of 3-bromoadamantane-1-carboxylic acid (10.0g,38.6mmol) in THF (25mL) was added slowly a solution of 1M BH3-THF in THF (115mL,3eq) and the mixture was stirred for 48 h. After addition of methanol and stirring for 30 min, purification by column chromatography (silica gel, heptane and MTBE as eluent) gave the desired product (8.37g, 88%).

¹H NMR(400MHz,DMSO-d₆):δppm 4.50(t,1H),3.02(d,2H),2.28/2.21(dm+dm,4H),2.11(m,2H),2.07(s,2H),1.66/1.56(dm+dm,2H),1.48/1.39(dm+dm,4H)；¹³C NMR(100MHz,DMSO-d₆)δppm 70.9,69.3,51.3,49.0,40.6,37.3,35.1,32.3。

And B:1- [ (3-bromo-1-adamantyl) methyl group]Pyrazoles

To a solution of the product of step A (8.37g,34.1mmol), 1H-pyrazole (2.79g,1.2eq) in toluene (100mL) was added (cyanomethylene) tributylphosphine (10.7mL,1.2eq) and the reaction mixture was stirred at 90 ℃ for 2H. Purification by column chromatography (silica gel, heptane and MTBE as eluent) gave the desired product (8.50g, 84%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.63(dd,1H),7.43(dd,1H),6.23(t,1H),3.87(s,2H),2.24/2.13(m+m,4H),2.1(m,2H),2.07(s,2H),1.63/1.50(m+m,2H),1.47/1.43(m+m,4H)；¹³C NMR(100MHz,DMSO-d₆)δppm 138.9,131.7,105.1,68.0,61.8,51.8,48.5,39.8,38.3,34.6,32.1；HRMS-ESI(m/z):[M+H]+C₁₄H₂₀BrN₂295.0810 found 295.0804.

Step C1- [ (3-bromo-1-adamantyl) methyl group]-5-methyl-pyrazole

To the product of step B (1.70g,5.76mmol) in THF (30mL) was added butyllithium (2.5M in THF, 12mL,5 eq.) at-78 ℃. Methyl iodide (7.2mL,5eq) was added to the mixture after 1 hour. After 10 minutes, the reaction mixture was washed with saturated NH₄The Cl solution was quenched, extracted with EtOAc and the combined organic layers were dried and concentrated to give the desired product (2.0g, 112%) which was used in the next step without further purification.

¹H NMR(400MHz,DMSO-d₆):δppm 7.31(d,1H),6.01(d,1H),3.76(s,2H),2.25/2.15(d+d,4H),2.24(s,3H),2.16(s,2H),2.10(m,2H),1.63/1.52(d+d,2H),1.52/1.49(d+d,4H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.2,138.0,105.2,68.2,58.3,52.1,48.5,40.5,38.4,34.5,32.2,11.8；HRMS-ESI(m/z):[M+H]+C₁₅H₂₂BrN₂309.0966 found value of 309.0962.

Step D2- [ [3- [ (5-methylpyrazol-1-yl) methyl ] methyl]-1-adamantyl]Oxy radical]Ethanol

A mixture of the product of step C (2.00g,6.47mmol), ethylene glycol (14.4mL,40eq) and DIPEA (5.6mL,5eq) was stirred at 120 ℃ for 6 h. After dilution with water and extraction with EtOAc, the combined organic phases were purified by column chromatography (silica gel, heptane and MTBE as eluent) to give the desired product (1.62g, 86.6%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.28(d,1H),5.99(m,1H),4.46(t,1H),3.75(s,2H),3.40(m,2H),3.32(m,2H),2.23(brs,3H),2.13(m,2H),1.61/1.52(m+m,4H),1.47/1.43(m+m,2H),1.45(s,2H),1.44-1.35(m,4H)；¹³C NMR(100MHz,DMSO-d₆)δppm 137.8,105.1,61.8,61.5,59.0,44.6,40.8,39.6,35.7,30.0,11.9；HRMS-ESI(m/z):[M+H]+C₁₇H₂₇N₂O₂291.2073 found value of 291.2069.

Step E:tert-butyl- [2- [ [3- [ (5-methylpyrazol-1-yl) methyl ] methyl]-1-adamantyl]Oxy radical]Ethoxy radical]-diphenyl-silane

To a solution of the product of step D (6.52g,22.5mmol) and imidazole (2.29g,1.5eq) in DCM (67mL) was added tert-butyl-chloro-diphenyl-silane (6.9mL,1.2eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, heptane and MTBE as eluent) gave the desired product (11.0g, 92.7%). LC/MS (C) ₃₃H₄₅N₂O₂Si)529[M+H]⁺。

Step FTert-butyl- [2- [ [3- [ (4-iodo-5-methyl-pyrazol-1-yl) methyl ] methyl]-1-adamantyl]Oxy radical]Ethoxy radical]-diphenyl-silane

To the product of step E (11.0g,20.8mmol) in DMF (105mL) was added N-iodosuccinimide (5.85g,1.25eq.) and the reaction mixture was stirred for 3 hours. After the reaction mixture was diluted with water and extracted with DCM, the combined organic phases were washed with saturated sodium thiosulfate and brine, dried and evaporated to give the desired product (11.0g, 81%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.70-7.36(m,10H),7.44(s,1H),3.86(s,2H),3.67(t,2H),3.45(t,2H),2.24(s,3H),2.12(m,2H),1.66-1.32(m,12H),0.98(s,9H)¹³C NMR(100MHz,DMSO-d₆)δppm 142.4,140.9,64.4,61.4,60.4,60.3,30.0,27.1,12.2；HRMS-ESI(m/z):[M+H]+C₃₃H₄₄IN₂O₂Calculated value of Si 655.2217 found value is 655.2217.

Step GTert-butyl- [2- [ [3- [ [ 5-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl]Methyl radical]-1-adamantyl]Oxy radical]Ethoxy radical]-diphenyl-silane

To the product of step F (11.0g,16.8mmol) in THF (84mL) was added chloro (isopropyl) magnesium-LiCl (1.3M in THF, 17mL,1.2eq) at 0 deg.C and the reaction mixture was stirred for 40 minutes, treated with 2-isopropoxy-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (10.3mL,3eq) and stirred for 10 minutes. With saturated NH₄After dilution with Cl solution and extraction with EtOAc, the combined organic phases were concentrated and purified by column chromatography (silica gel, heptane and MTBE as eluent) to give the desired product (9.0g, 82%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.66(d,4H),7.47(s,1H),7.45(t,2H),7.40(t,4H),3.77(s,2H),3.67(t,2H),3.44(t,2H),2.36(s,3H),2.11(br,2H),1.60/1.48(d+d,4H),1.44(d,2H),1.44(s,2H),1.40(d,4H),1.23(s,12H),0.97(s,9H)；¹³C NMR(100MHz,DMSO-d₆)δppm146.9,144.2,133.8,130.2,128.3,125.7,104.6,83.0,72.5,64.4,61.4,58.9,44.6,40.7,39.6,38.7,35.6,30.0,27.1,25.2,19.3,12.1；HRMS-ESI(m/z):[M+H]+C₃₉H₅₆BN₂O₄Calculated Si 655.4102 found 655.4108.

Preparation example 103-bromo-6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-l) propylamino group]Pyridine-2-carboxylic acid methyl ester

Step A6- [ bis (tert-butoxycarbonyl) amino group]-3-bromo-pyridine-2-carboxylic acid methyl ester

To a solution of 6-amino-3-bromo-pyridine-2-carboxylic acid methyl ester (25.0g,108.2mmol) and DMAP (1.3g,0.1eq) in DCM (541mL) at 0 deg.C was added Boc₂O (59.0g,2.5eq) and the reaction mixture was stirred for 2.5 hours. Adding saturated NaHCO₃After the solution was extracted with DCM, the combined organic phases were dried and concentrated to give the desired product (45.0g, 72.3%). LC/MS (C)₁₇H₂₃BrN₂O₆Na)453[M+Na]⁺。

And B:3-bromo-6- (tert-butoxycarbonylamino) pyridine-2-carboxylic acid methyl ester

To a solution of the product of step A (42.7g,74.34mmol) in DCM (370mL) was added TFA (17.1mL,3eq) at 0 deg.C and the reaction mixture was stirred for 18 h. With saturated NaHCO₃After washing the solution with brine, the combined organic phases were dried, concentrated and purified by column chromatography (silica gel, heptane and EtOAc as eluent) to give the desired product (28.3g, 115.2%).

¹H NMR(400MHz,DMSO-d₆):δppm 10.29(s,1H),8.11(d,1H),7.88(d,1H),3.87(s,3H),1.46(s,9H)¹³C NMR(100MHz,DMSO-d₆)δppm 165.6,153.1,151.8/148.3,143.5,116.3,109.2,53.2,28.4.LC/MS(C₁₂H₁₅BrN₂O₄Na)353[M+Na]⁺。

Step C3-bromo-6- [ tert-butoxycarbonyl- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propyl ] methyl-l-butanamido]Amino group]Pyridine-2-carboxylic acid methyl ester

To a solution of the product of step B (10.0g,30.1967mmol) in acetone (150mL) was added Cs ₂CO₃(29.5g,3eq) and 3, 6-dichloro-4- (3-iodopropyl) -5-methyl-pyridazine (9.9g,1eq) and the reaction mixture was stirred for 18 h. After dilution with water and extraction with EtOAc, the combined organic phases were washed with brine, dried and concentrated to give the desired product (17.5g, 108%).

¹H NMR(400MHz,DMSO-d₆):δppm 8.13(d,1H),7.78(d,1H),3.91(t,2H),3.89(s,3H),2.79(m,2H),2.38(s,3H),1.82(m,2H),1.46(s,9H)；¹³C NMR(100MHz,DMSO-d₆)δppm 165.3,157.6,156.6,153.2,152.9,147.2,143.1,142.2,139.7,122.6,111.8,82.2,53.3,46.4,28.1,27.7,26.5,16.3；HRMS-ESI(m/z):[M+Na]⁺C₂₀H₂₃BrCl₂N₄NaO₄555.0177; found 555.0172.

Step D:3-bromo-6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Pyridine-2-carboxylic acid methyl ester

A solution of the product of step C (17.5g,32.7mmol) in 1,1,1,3,3, 3-hexafluoroisopropanol (330mL) was stirred at 110 ℃ for 18 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluent) afforded the desired product (9.9g, 70%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.63(d,1H),7.22(t,1H),6.57(d,1H),3.83(s,3H),3.30(m,2H),2.83(m,2H),2.37(s,3H),1.74(m,2H)¹³C NMR(100MHz,DMSO-d₆)δppm 166.5,141.5,112.6,52.9,40.9,28.0,27.0,16.4。

Preparation example 11(4-methoxyphenyl) methyl 3-bromo-6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Pyridine-2-carboxylic acid esters

Step A:3-bromo-6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Pyridine-2-carboxylic acid

The product of preparation 10 (35.39g,81.52mmol) and LiOH XH₂A mixture of O (13.68g,4eq) in 1, 4-dioxane (408mL) and water (82mL) was stirred at 60 ℃ for 1 hour. After quenching the reaction with 1M HCl solution and extraction with EtOAc, the combined organic phases were dried, concentrated, and purified by flash chromatography (silica gel, using DCM and MeOH as eluents) to give the desired product (27.74g, 81%).

LC/MS(C₁₄H₁₄BrCl₂N₄O₂)421[M+H]⁺。

And B, step B:(4-methoxyphenyl) methyl 3-bromo-6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]2-Pyridinecarboxylic acid ester

To the product of step A (27.7g,65.9mmol), (4-methoxyphenyl) methanol (16.4mL,2eq) and PPh₃(34.6g,2eq) in toluene (660mL) and THF (20mL) was added dropwise diisopropyl azodicarboxylate (26mL,2eq) and the reaction mixture was stirred at 50 ℃ for 1 hour.Purification by flash chromatography (silica gel, using heptane and EtOAc as eluent) afforded the desired product (23.65g, 66.4%).

¹H NMR(500MHz,dmso-d6)δppm 7.62(d,1H),7.37(dn,2H),7.21(t,1H),6.91(dm,2H),6.56(d,1H),5.25(s,2H),3.74(s,3H),3.30(q,2H),2.81(m,2H),2.33(s,3H),1.73(m,2H)；¹³C NMR(500MHz,dmso-d6)δppm 165.9,159.7,157.6,157.5,156.8,148.0,142.7,141.5,139.7,130.6,127.8,114.3,112.6,101.6,67.0,55.6,40.9,28.0,27.1,16.4；HRMS-ESI(m/z):[M+H]+C₂₂H₂₂BrCl₂N₄O₃539.0252; found 539.0246.

Preparation example 126- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- [2- (p-tolylsulfonyloxy) ethoxy ] ethoxy]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

Step A6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino group]-3- [ 5-methyl-1- [ [3- [2- [ tert-butyl (diphenyl) silyl ] silane]Oxoethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

The product of preparation 10 (15.0g,34.55mmol), the product of preparation 7 (30.7g,1.3eq), Cs₂CO₃(33.8g,3.0eq) and Pd (AtaPhos) ₂Cl₂(1.53g,0.1eq) in 1, 4-dioxane (207mL) and H₂The mixture in O (34.5mL) was stirred at 80 ℃ for 1.5 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluent) afforded the desired product (18.5g, 58%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.69-7.37(m,10H),7.32(d,1H),7.23(s,1H),6.98(t,1H),6.63(d,1H),3.82(s,2H),3.67(t,2H),3.58(s,3H),3.46(t,2H),3.35(m,2H),2.86(m,2H),2.40(s,3H),2.06(s,3H),1.78(m,2H),1.35(s,2H),1.27/1.2(m+m,4H),1.15/1.09(m+m,4H),1.05/0.97(m+m,2H),0.97(s,9H),0.84(s,6H)；HRMS-ESI(m/z):[M+H]+C₅₀H₆₃Cl₂N₆O₄Calculated value of Si 909.4057; found 909.4053.

Step B6- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -3- [ 5-methyl-1- [ [ [2- [ tert-butyl (diphenyl) silyl ] silane]Oxoethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

The product of step A (18.5g,20.3mmol), Cs₂CO₃(13.2g,2eq), DIPEA (7.1mL,2eq), and Pd (Atapos)₂Cl₂A mixture of (900mg,0.1eq) in 1, 4-dioxane (102mL) was stirred at 110 ℃ for 18 h. After filtration and concentration, the residue was dissolved in DCM, washed with water and purified by column chromatography (silica gel, DCM and EtOAc as eluent) to give the desired product (12.6g, 71%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.85(d,1H),7.69(d,1H),7.66(dm,4H),7.47-7.36(m,6H),7.38(s,1H),3.97(t,2H),3.87(s,2H),3.68(t,2H),3.66(s,3H),3.47(t,2H),2.87(t,2H),2.30(s,3H),2.14(s,3H),1.99(br.,2H),1.38(s,2H),1.32-0.96(br.,10H),0.98(s,9H),0.85(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.9,137.6,120.5,64.4,61.7,58.9,52.3,46.0,43.4,30.2,27.1,24.6,21.0,15.5,10.9；HRMS-ESI(m/z):[M+H]+C₅₀H₆₂ClN₆O₄Calculated value of Si 873.4290; found 873.4291.

Step C6- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -3- [1- [ [3- (2-hydroxyethoxy) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

To the product of step B (8.46g,9.68mmol) in THF (95mL) was added a 1M solution of TBAF in THF (10.6mL,1.1eq) at 0 deg.C and the reaction mixture was stirred for 2 h. With saturated NH ₄After quenching the reaction with Cl solution and extraction with EtOAc, the combined organic phases were washed with brine, dried and purified by column chromatography (silica gel, DCM and MeOH as eluent) to give the desired product (5.38g, 88%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.86(d,1H),7.71(d,1H),7.38(s,1H),4.46(t,1H),3.97(t,2H),3.87(s,2H),3.70(s,3H),3.40(m,2H),3.35(t,2H),2.87(t,2H),2.30(s,3H),2.15(s,3H),1.99(m,2H),1.42-0.95(m,12H),0.87(s,6H)；HRMS-ESI(m/z):[M+H]+C₃₄H₄₄ClN₆O₄635.3113; found 635.3112.

Step D6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-3- [1- [ [3- (2-hydroxyethoxy) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

Using Buchwald general procedure I at 130 ℃ for 1 hour, 3.7g of the product from step C (5.78mmol) and 1.74g of 1, 3-benzothiazol-2-amine (2eq) were used as starting materials to give 3.1g of the desired product (72% yield).

¹H NMR(400MHz,DMSO-d₆):δppm 7.96(d,1H),7.82(br.,1H),7.70(d,1H),7.50(br.,1H),7.38(s,1H),7.35(t,1H),7.17(t,1H),4.46(br.,1H),4.00(t,2H),3.88(s,2H),3.70(s,3H),3.40(brt.,2H),3.35(t,2H),2.86(t,2H),2.32(s,3H),2.16(s,3H),2.03-1.94(m,2H),1.42-0.96(m,12H),0.87(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.8,137.5,126.4,122.4,122.1,119.0,62.1,61.5,59.0,52.6,45.4,30.2,24.3,21.7,12.6,10.9；HRMS-ESI(m/z):[M+H]+C₄₁H₄₉N₈O₄749.3597 as the calculated value of S; found 749.3595.

Step E6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- [2- (p-tolylsulfonyloxy) ethoxy ] ethoxy]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

To a solution of the product of step D (3.85g,5.14mmol) and triethylamine (2.15mL,3eq) in DCM (50mL) was added p-tolylsulfonyl 4-methylbenzenesulfonate (2.51g,1.5eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, heptane and EtOAc as eluent) afforded the desired product (3.2g, 69%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.96(d,1H),7.81(br.,1H),7.77(d,2H),7.70(d,1H),7.50(br.,1H),7.46(d,2H),7.39(s,1H),7.35(t,1H),7.17(t,1H),4.06(t,2H),4.00(t,2H),3.85(s,2H),3.69(s,3H),3.49(t,2H),2.86(t,2H),2.40(s,3H),2.32(s,3H),2.15(s,3H),1.99(m,2H),1.32-0.93(m,12H),0.84(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.8,137.6,130.6,128.1,126.4,122.4,122.1,119,71.5,58.8,58.4,52.6,45.4,30.1,24.3,21.7,21.6,12.6,10.9；HRMS-ESI(m/z):[M+H]+C₄₈H₅₅N₈O₆S₂903.3686 found 903.3685.

Preparation example 13:(4-methoxyphenyl) methyl 6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- [3- (p-tolylsulfonyloxy) propyl ] group]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid esters

Step A(4-methoxyphenyl) methyl 3- [1- [ [3- [3- [ tert-butyl (diphenyl) silyl)]Oxopropyl radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Pyridine-2-carboxylic acid esters

The product of preparation 11 (3.67g,6.79mmol), the product of preparation 8 (5.09g,1.1eq), Pd (AtaPhos)₂Cl₂(301mg,0.1eq) and Cs₂CO₃(6.64g,3eq) in 1, 4-dioxane (41mL) and H₂The mixture in O (6.8mL) was stirred at 80 ℃ for 18 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluent) afforded the desired product (4.43g, 64%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.62-7.38(m,10H),7.32(d,1H),7.26(s,1H),7.10(m,2H),6.98(t,1H),6.83(m,2H),6.63(d,1H),4.98(s,2H),3.74(s,2H),3.70(s,3H),3.58(t,2H),3.35(m,2H),2.84(m,2H),2.34(s,3H),2.02(s,3H),1.77(m,2H),1.43(m,2H),1.18-0.85(m,12H),1.09(t,2H),0.97(s,9H),0.77(s,6H)；HRMS-ESI(m/z):[M+H]+C₅₈H₇₁Cl₂N₆O₄Calculated value of Si 1013.4683; measured value 1013.4683;

step B(4-methoxyphenyl) methyl 3- [1- [ [3- [3- [ tert-butyl (diphenyl) silyl)]Oxopropyl radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c) ]Pyridazin-8-yl) pyridines2-formic acid ester

The product of step A (4.43g,4.37mmol), Cs₂CO₃(2.84g,2eq), DIPEA (1.5mL,2eq), and Pd (Atapos)₂Cl₂A mixture of (193mg,0.1eq) in 1, 4-dioxane (22mL) was stirred at 110 ℃ for 18 h. After quenching the reaction with water and extraction with EtOAc, the combined organic phases were dried, concentrated and purified by column chromatography (silica gel, DCM and EtOAc as eluent) to give the desired product (2.83g, 66%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.84(d,1H),7.68(d,1H),7.59(d,4H),7.44(t,2H),7.42(t,4H),7.38(s,1H),7.14(d,2H),6.87(d,2H),5.07(s,2H),3.96(t,2H),3.78(s,2H),3.71(s,3H),3.59(t,2H),2.86(t,2H),2.29(s,3H),2.08(s,3H),1.97(qn,2H),1.43(qn,2H),1.12(s,4H),1.10(s,2H),1.09(t,2H),0.97(s,9H),0.95(s,2H),0.94/0.91(d+d,4H),0.78(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 166.9,159.6,156.3,153.6,150.8,147.7,140.1,137.5,137.3,136.0,135.5,133.8,130.3,130.1,129.1,128.3,127.6,123.1,120.5,115.5,114.3,66.8,64.8,64.8,59.6,55.6,50.5,48.1,46.4,46.0,44.2,39.3,38.1,31.7,30.6,27.2,26.1,24.6,21.0,19.3,15.5,10.9；

HRMS-ESI(m/z):[M+H]+C₅₈H₇₀ClN₆O₄Calculated value of Si 977.4916; found 977.4915.

And C:(4-methoxyphenyl) methyl 6- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -3- [1- [ [3- (3-hydroxypropyl) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid esters

To the product of step B (2.83g,2.89mmol) in THF (95mL) was added a 1M solution of TBAF in THF (3.2mL,1.1eq) at 0 deg.C and the reaction mixture was stirred for 2 h. With saturated NH₄After quenching the reaction with Cl solution and extraction with EtOAc, the combined organic phases were washed with brine, dried, concentrated and purified by column chromatography (silica gel, DCM and MeOH as eluent) to give the desired product (2.21g, 103%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.85(d,1H),7.70(d,1H),7.39(s,1H),7.17(d,2H),6.90(d,2H),5.09(s,2H),4.34(t,1H),3.96(t,2H),3.79(s,2H),3.74(s,3H),3.32(q,2H),2.86(t,2H),2.29(s,3H),2.09(s,3H),1.98(qn,2H),1.34(qn,2H),1.13(s,2H),1.13(s,4H),1.06(t,2H),0.99/0.95(d+d,4H),0.97(s,2H),0.78(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm166.9,159.7,156.4,153.6,150.8,147.7,140.2,137.5,137.3,136.0,130.2,129.1,127.6,123.1,120.4,115.5,114.3,66.8,66.8,62.1,59.7,55.6,50.6,48.2,46.5,46.0,44.3,39.7,38.1,31.8,30.6,26.5,24.6,21.0,15.5,10.9；HRMS-ESI(m/z):[M+H]+C₄₂H₅₂ClN₆O₄739.3739; found 739.3739.

Step D(4-methoxyphenyl) methyl 6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] ]Pyridazin-8-yl radicals]-3- [1- [ [3- (3-hydroxypropyl) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]2-Pyridinecarboxylic acid ester

The product of step C (1.71g,2.31mmol), 1, 3-benzothiazol-2-amine (695mg,2eq), Pd₂dba₃A mixture of (212mg,0.1eq), XantPhos (268mg,0.2eq) and DIPEA (1.2mL,3eq) in cyclohexanol (14mL) was stirred at 130 ℃ for 1 hour. Purification by column chromatography (silica gel, heptane, DCM and MeCN as eluents) gave the desired product (1.25g, 63%).

¹H NMR(400MHz,DMSO-d₆):δppm 12.08/10.87(brs/brs,1H),7.95(d,1H),7.81(br,1H),7.68(d,1H),7.50(br,1H),7.39(s,1H),7.35(t,1H),7.18(d,2H),7.17(t,1H),6.90(d,2H),5.10(s,2H),4.34(t,1H),3.99(t,2H),3.79(s,2H),3.74(s,3H),3.33(q,2H),2.85(t,2H),2.32(s,3H),2.11(s,3H),1.98(qn,2H),1.34(qn,2H),1.14(s,4H),1.14(s,2H),1.07(t,2H),1.00/0.95(d+d,2H),0.99/0.95(d+d,4H),0.79(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 140.0,137.6,130.2,126.4,122.4,122.0,119.0,114.3,66.7,62.1,59.6,55.6,50.6,48.2,46.5,45.4,44.3,39.7,30.6,26.5,24.3,21.7,12.6,11.0；HRMS-ESI(m/z):[M+H]+C₄₉H₅₇N₈O₄853.4223 as the calculated value of S; found 853.4229.

Step E:(4-methoxyphenyl) methyl 6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-67-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- [3- (p-tolylsulfonyloxy) propyl ] group]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid esters

To a solution of the product of step D (1.25g,1.47mmol) and triethylamine (0.61mL,3eq) in DCM (15mL) was added p-tolylsulfonyl 4-methylbenzenesulfonate (717mg,1.5eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, heptane and EtOAc as eluent) afforded 800mg (54%) of the desired product.

¹H NMR(400MHz,DMSO-d₆):δppm 7.95(d,1H),7.88(brs,1H),7.77(m,2H),7.68(d,1H),7.62(brs,1H),7.47(m,2H),7.39(s,1H),7.35(brs,1H),7.17(brs,1H),7.10(m,2H),6.90(m,2H),5.09(s,2H),4.00(m,2H),3.98(t,2H),3.77(s,2H),3.74(s,3H),2.85(t,2H),2.40(s,3H),2.32(s,3H),2.09(s,3H),1.98(m,2H),1.45(m,2H),1.17-0.8(m,12H),0.98(m,2H),0.77(s,6H)；HRMS-ESI(m/z):[M+H]+C₅₆H₆₃N₈O₆S₂1007.4312; found 1007.4318.

Preparation example 14(4-methoxyphenyl) methyl 6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-3- [ 5-methyl-1- [ [3- [2- (p-tolylsulfonyloxy) ethoxy ] ethoxy]-1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid esters

Step A(4-methoxyphenyl) methyl 3- [1- [ [3- [2- [ tert-butyl (diphenyl) silyl)]Oxoethoxy radical]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Pyridine-2-carboxylic acid esters

The product of preparation 11 (3.67g,6.79mmol), the product of preparation 9 (4.89g,1.1eq), Pd (AtaPhos)₂Cl₂(301mg,0.1eq) and Cs₂CO₃(6.64g,3eq) in 1, 4-dioxane (41mL) and H₂The mixture in O (6.8mL) was stirred at 80 ℃ for 12 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluent) afforded the desired product (3.0g, 45%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.69-7.37(m,10H),7.31(d,1H),7.24(s,1H),7.12(m,2H),6.98(t,1H),6.83(m,2H),6.62(d,1H),4.99(s,2H),3.76(s,2H),3.70(s,3H),3.66(t,2H),3.45(t,2H),3.35(m,2H),2.85(m,2H),2.34(s,3H),2.12(m,2H),2.02(s,3H),1.77(m,2H),1.65-1.33(m,12H),0.97(s,9H)；HRMS-ESI(m/z):[M+H]+C₅₅H₆₅Cl₂N₆O₅Calculated value of Si 987.4163; found 987.4158.

Step B(4-methoxyphenyl) methyl 3- [1- [ [3- [2- [ tert-butyl (diphenyl) silyl)]Oxoethoxy radical]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) pyridine-2-carboxylic acid esters

The product of step A (3.00g,3.00mmol), Cs ₂CO₃(1.95g,2eq), DIPEA (1.0mL,2eq), and Pd (Atapos)₂Cl₂A mixture of (212mg,0.1eq) in 1, 4-dioxane (15mL) was stirred at 110 ℃ for 18 h. Purification by column chromatography (silica gel, DCM and MeOH as eluents) gave the desired product (1.74g, 60%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.84(d,1H),7.68(d,1H),7.68-7.37(m,10H),7.36(s,1H),7.16(m,2H),6.87(m,2H),5.08(s,2H),3.96(m,2H),3.81(s,2H),3.72(s,3H),3.67(t,2H),3.46(t,2H),2.87(t,2H),2.29(s,3H),2.13(m,2H),2.09(s,3H),1.98(m,2H),1.65-1.37(m,12H),0.97(s,9H)；HRMS-ESI(m/z):[M+H]+C₅₅H₆₄ClN₆O₅Calculated value of Si 951.4396; found 951.4397.

Step C(4-methoxyphenyl) methyl 6- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -3- [1- [ [3- (2-hydroxyethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid esters

To the product of step B (1.73g,1.82mmol) in THF (20mL) was added 1M TBAF in THF (2.0mL,1.1eq) at 0 deg.C and the reaction mixture was stirred for 2 h. Purification by column chromatography (silica gel, DCM and MeOH as eluent) gave the desired product (1.06g, 82%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.85(d,1H),7.71(d,1H),7.36(s,1H),7.19(m,2H),6.90(m,2H),5.10(s,2H),4.47(t,1H),3.96(m,2H),3.81(s,2H),3.75(s,3H),3.40(m,2H),3.34(t,2H),2.87(t,2H),2.29(s,3H),2.14(m,2H),2.10(s,3H),1.98(m,2H),1.67-1.36(m,12H)；HRMS-ESI(m/z):[M+H]+C₃₉H₄₆ClN₆O₅713.3218; found 713.3217.

Step D(4-methoxyphenyl) methyl 6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-3- [1- [ [3- (2-hydroxyethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid esters

The product of step C (1.00g,1.40mmol), 1, 3-benzothiazol-2-amine (421mg,2eq), Pd₂dba₃A mixture of (128mg,0.1eq), XantPhos (162mg,0.2eq) and DIPEA (0.72mL,3eq) in cyclohexanol (10mL) was stirred at 130 ℃ for 1 hour. Purification by column chromatography (silica gel, heptane, then DCM and MeOH as eluent) gave the desired product (600mg, 53%).

¹H NMR(400MHz,DMSO-d₆):δppm 12.18/10.84(brs/brs,1H),7.94(d,1H),7.83(br,1H),7.69(d,1H),7.57(br,1H),7.36(s,1H),7.35(brt,1H),7.20(d,2H),7.17(brt,1H),6.91(d,2H),5.11(s,2H),4.47(brt,1H),4.00(t,2H),3.81(s,2H),3.75(s,3H),3.41(brq,2H),3.35(t,2H),2.85(t,2H),2.32(s,3H),2.14(m,2H),2.12(s,3H),1.99(qn,2H),1.62/1.53(d+d,4H),1.53(s,2H),1.49/1.44(d+d,2H),1.44(s,4H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.9,137.6,130.1,126.4,122.4,122.0,118.9,114.2,66.7,61.9,61.5,59.5,55.6,45.4,44.7,40.8,39.5,35.6,30.1,24.3,21.7,12.6,10.8；HRMS-ESI(m/z):[M+H]+C₄₆H₅₁N₈O₅827.3703 is the calculated value of S; found 827.3709.

Step E(4-methoxyphenyl) methyl 6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-3- [ 5-methyl-1- [ [3- [2- (p-tolylsulfonyloxy) ethoxy ] group]-1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid esters

To the product of step D (600mg,0.726mmol) and N, N-diethylethylamine (0.31mL,3eq) in dichloromethane (7mL) was added p-tolylSulfonyl 4-methylbenzenesulfonate (357mg,1.5eq) and the reaction mixture was stirred for 18 hours. Purification by flash chromatography (silica gel using DCM and MeOH as eluent) afforded 354mg (50%) of the desired product.¹H NMR(500MHz,dmso-d6)δppm 12.22/10.85(brs/brs,1H),7.94(d,1H),7.81(br,1H),7.77(d,2H),7.70(d,1H),7.52(br,1H),7.45(d,2H),7.37(s,1H),7.35(t,1H),7.19(d,2H),7.17(t,1H),6.89(d,2H),5.10(s,2H),4.05(t,2H),4.00(t,2H),3.79(s,2H),3.74(s,3H),3.49(t,2H),2.86(t,2H),2.40(s,3H),2.32(s,3H),2.11(m,2H),2.11(s,3H),1.99(qn,2H),1.55-1.36(m,12H)；¹³C NMR(500MHz,dmso-d6)δppm 139.9,137.6,130.5,130.3,128.1,126.4,122.4,122.0,118.9,114.2,71.4,66.8,59.4,58.2,55.6,45.4,30.0,24.2,21.6,21.6,12.6,10.9；HRMS-ESI(m/z):[M+H]+C₅₃H₅₇N₈O₇S₂981.3792; found 981.3795.

Preparation example 15:2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid ethyl ester

Step A2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid

The product of preparation 3a (35.39g,81.52mmol) and LiOH XH₂A mixture of O (4eq) in 1, 4-dioxane (408mL) and water (82mL) was stirred at 60 ℃ for 1 hour. After quenching the reaction with 1M HCl solution and extraction with EtOAc, the combined organic phases were dried, concentrated, and purified by flash chromatography (silica gel, using DCM and MeOH as eluents) to give the desired product (27.7g, 81%).

¹H NMR(500MHz,dmso-d6)δppm 7.56(dd,1H),7.43(brd.,1H),6.96(t,1H),4.18(t,2H),4.05(t,2H),3.28(t,2H),2.84(t,2H),2.29(s,3H),2.07(m,2H),1.97(m,2H)；¹³C NMR(500MHz,dmso-d6)δppm 166.4,154.8,152.1,151.8,151.1,147.1,143.9,135.7,134.0,133.8,129.0,124.9,117.6,82.3,68.8,46.3,31.0,24.0,22.5,19.8,15.7；HRMS-ESI(m/z):[M+H]+C₂₁H₂₀ClFIN₄O₃588.9973 is the calculated value of S; found 588.9969.

Step B2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid ethyl ester

To the product of step A (27.7g,65.9mmol), ethanol (2eq) and PPh₃(2eq) to a mixture of toluene (660mL) and THF (20mL) was added dropwise diisopropyl azodicarboxylate (2eq) and the reaction was stirred at 50 ℃ for 1 hour. Purification by flash chromatography (silica gel, using heptane and EtOAc as eluent) afforded the desired product (23.65g, 66.4%).

¹H NMR(500MHz,dmso-d6)δppm 7.59(dd,1H),7.44(dm,1H),6.98(t,1H),4.29(m,2H),4.25(q,2H),4.08(t,2H),3.24(t,2H),2.89(t,2H),2.32(s,3H),2.09(m,2H),2.04(m,2H),1.28(t,3H)；¹³C NMR(500MHz,dmso-d6)δppm 162.6,155.4,152.2,151.7,151.3,147.0,134.0,124.9,117.6,82.4,68.3,60.7,46.3,30.8,24.1,23.1,19.7,15.7,14.6；HRMS-ESI(m/z):[M+H]⁺C₂₃H₂₄ClFIN₄O₃Calculated value of S is 617.0286, found value is 617.0282.

Example 12- {6- [ (1, 3-benzothiazol-2-yl) amino]-1,2,3, 4-tetrahydroquinolin-1-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- (6-bromo-1, 2,3, 4-tetrahydroquinolin-1-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of benzoyl isothiocyanate (380 μ L,2.83mmol,1.2eq) in acetone (10mL) was added 6-bromo-1, 2,3, 4-tetrahydroquinoline (500mg,2.36mmol,1eq) and the mixture was heated at reflux for 1 hour. The mixture was poured onto ice water and the precipitate was filtered, washed with water and dried to give a light yellow solid. The solid was added to 1N aqueous sodium hydroxide (10mL) and the suspension was heated at 80 ℃ for 30 minutes, cooled to room temperature and poured onto cold 1N aqueous hydrochloric acid. The pH was adjusted to pH 8 with saturated aqueous sodium carbonate solution, the solid was collected by filtration and washed with water to give a yellow solid. The solid was combined with ethyl bromopyruvate (296. mu.L, 2.36mmol,1eq) in ethyl acetate The alcohol (10mL) suspension was heated at reflux for 2 hours. The mixture was cooled to room temperature and then partitioned between ethyl acetate and water. The aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% ethyl acetate in isoheptane to give the desired product as a yellow gum (232mg,0.63mmol, 27%).

LC/MS(C₁₅H₁₅BrN₂O₂S)367[M+H]⁺；RT 1.42(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.99(d,1H),7.87(s,1H),7.53-7.48(m,1H),7.44-7.38(m,1H),4.27(q,2H),3.83(t,2H),2.83-2.73(m,2H),2.00-1.90(m,2H),1.29(t,3H)。

Step B2- {6- [ (1, 3-benzothiazol-2-yl) amino]-1,2,3, 4-tetrahydroquinolin-1-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step A (93.8mg,0.26mmol,1eq), 2-aminobenzothiazole (46.0mg,0.31mmol,1.2eq), cesium carbonate (166mg,0.51mmol,2eq) and 1, 4-dioxane (4mL) and the mixture was purged with nitrogen (10min), then Brettphos (13.7mg,0.03mmol,0.1eq) and tris (dibenzylideneacetone) dipalladium (0) (23.4mg,0.03mmol,0.1eq) were added and heated at 120 ℃ for 2 hours under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMCartridge silica) and gradient eluted with 0-20% ethyl acetate in isoheptane to give the desired product as a yellow gum (85.1mg,0.19mmol, 76%).

LC/MS(C₂₂H₂₀N₄O₂S₂)437[M+H]⁺；RT 1.40(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.87(d,1H),7.83-7.78(m,1H),7.69-7.65(m,1H),7.63-7.57(m,2H),7.36-7.29(m,1H),7.18-7.13(m,1H),7.03-6.98(m,1H),4.28(q,J＝7.08Hz,2H),3.92-3.84(m,2H),2.80(t,J＝6.30Hz,2H),2.00-1.89(m,2H),1.30(t,J＝7.09Hz,2H)。

Step C2- {6- [ (1, 3-benzothiazol-2-yl) amino]-1,2,3, 4-tetrahydroquinolin-1-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (85.1mg,0.19mmol,1eq) in tetrahydrofuran (2mL) and methanol (1mL) was added 1N aqueous sodium hydroxide (0.39mL,0.39mmol,2eq) and the mixture was heated at 50 ℃ for 3 h. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 30mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-6% methanol in dichloromethane afforded the product which was further purified by preparative HPLC (HPLC-V-a2) to afford the desired product as a cream solid (1.2mg, 1.5%).

HRMS-ESI(m/z)[M+H]+C₂₀H₁₇N₄O₂S₂409.0793, found 409.0830

Example 22- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-indol-1-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- (5-bromo-1H-indol-1-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a cooled solution of 5-bromoindole (150mg,0.77mmol,1eq) in dimethylformamide (2mL) was added sodium hydride (60% dispersion; 36.7mg,1.53mmol,2eq) portionwise and the mixture was stirred at 0 ℃ for 30 minutes, then methyl 2-chloro-4-thiazolecarboxylate (272mg,1.53mmol,2eq) was added, then warmed to room temperature and stirred overnight. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3x30mL), the combined organic extracts were washed with brine (3x30mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel tube) Purification, eluting with a gradient of 0-20% ethyl acetate in isoheptane afforded the desired product as a yellow solid (72.8mg,0.22mmol, 28%).

LC/MS(C₁₃H₉BrN₂O₂S)339[M+H]⁺；RT 1.35(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.41-8.38(m,1H),8.37(s,1H),8.02(d,J＝3.5Hz,1H),7.93(d,J＝2.0Hz,1H),7.57(dd,J＝8.8,2.0Hz,1H),6.86(dd,J＝3.5,0.8Hz,1H),3.89(s,3H)。

Step B2- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-indol-1-yl } -1, 3-thiazole-4-carboxylic acid methyl ester

To an oven dried microwave vial was added the product of step A (72.8mg,0.22mmol,1eq), 2-aminobenzothiazole (38.9mg,0.26mmol,1.2eq), cesium carbonate (141mg,0.43mmol,2eq) and 1, 4-dioxane (2mL) and the mixture was purged with nitrogen (10min), followed by Brettphos (11.6mg,0.02mmol,0.1eq) and tris (dibenzylideneacetone) dipalladium (0) (19.8mg,0.02mmol,0.1eq) and heated at 120 ℃ for 2 hours under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 30mL), the combined organic extracts were washed with brine (30mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-20% ethyl acetate in isoheptane to afford the desired product as a pale yellow solid (10.5mg,0.03mmol, 12%).

LC/MS(C₂₀H₁₄N₄O₂S₂)407[M+H]⁺；RT 1.35(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.39(d,J＝8.9Hz,1H),8.34(d,J＝4.4Hz,1H),7.96(d,J＝3.6Hz,1H),7.84-7.78(m,1H),7.67-7.57(m,2H),7.39-7.30(m,2H),7.20-7.12(m,1H),6.92(d,J＝3.7Hz,1H),3.90(s,3H)。

Step C2- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-indol-1-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (10.5mg,0mol,1eq) in tetrahydrofuran (2mL) and methanol (1mL) was added 1N aqueous sodium hydroxide (0.05mL,0.05mmol,2eq) and the mixture was heated at 50 ℃ for 2 h. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 30mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. Purification by preparative HPLC (HPLC-V-A2) gave the desired product as a cream-colored solid (3.5mg,0.01mmol, 35%).

HRMS-ESI(m/z)[M+H]+C₁₉H₁₃N₄O₂S₂393.0480, found 393.0503.

Example 32- {5- [ (1, 3-benzothiazol-2-yl) amino]-2, 3-dihydro-1H-indol-1-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- (5-bromo-2, 3-dihydro-1H-indol-1-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of benzoyl isothiocyanate (0.33mL,2.42mmol,1.2eq) in acetone (10mL) was added 5-bromoindoline (400mg,2.02mmol,1eq) and the mixture was heated at reflux for 1 hour. The reaction was poured onto ice water and the precipitate was filtered, washed with water and dried to give a pale yellow solid. The solid was added to 1N aqueous sodium hydroxide (10mL) and the suspension was heated at 80 ℃ for 30 minutes, cooled to room temperature and poured into cold aqueous 1N hydrochloric acid. The pH was adjusted to pH 8 with saturated aqueous sodium carbonate solution, the solid was collected by filtration and washed with water to give a yellow solid. A suspension of the solid and ethyl bromopyruvate (253. mu.L, 2.02mmol,1eq) in ethanol (10mL) was heated under reflux for 2 hours and then cooled to room temperature. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-20% ethyl acetate in isoheptane to give the desired product as a yellow solid (377mg,1.07mmol, 53%).

LC/MS(C₁₄H₁₃BrN₂O₂S)353[M+H]⁺；RT 1.39(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.00(d,J＝8.4Hz,1H),7.92(s,1H),7.57-7.53(m,1H),7.47-7.44(m,1H),4.30(q,J＝7.1Hz,2H),4.08(t,2H),3.34-3.26(m,2H),1.31(t,3H)。

Step B2- {5- [ (1, 3-benzothiazol-2-yl) amino]-2, 3-dihydro-1H-indol-1-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step A (100mg,0.28mmol,1eq), 2-aminobenzothiazole (51.0mg,0.34mmol,1.2eq), cesium carbonate (129mg,0.4mmol,2eq) and 1, 4-dioxane (2mL) and the mixture was purged with nitrogen (10min), followed by Brettphos (10.6mg,0.02mmol,0.1eq) and tris (dibenzylideneacetone) dipalladium (0) (18.2mg,0.02mmol,0.1eq) and then heated at 120 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a brown gum (29.8mg,0.07mmol, 36%).

LC/MS(C₂₁H₁₈N₄O₂S₂)423[M+H]⁺；RT 1.38(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.01(d,1H),7.87(s,1H),7.83-7.78(m,1H),7.61-7.57(m,1H),7.45(s,1H),7.33-7.28(m,1H),7.18-7.10(m,1H),7.00(td,J＝1.24,7.55Hz,1H),4.31(q,J＝7.12Hz,2H),4.13-4.06(m,2H),3.43-3.34(m,2H),1.33(t,J＝7.12Hz,3H)。

Step C2- {5- [ (1, 3-benzothiazol-2-yl) amino]-2, 3-dihydro-1H-indol-1-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (29.8mg,0.07mmol,1eq) in tetrahydrofuran (2mL) and methanol (1mL) was added 1N aqueous sodium hydroxide (0.14mL,0.14mmol,2eq) and the mixture was heated at 50 ℃ for 1 h. Placing the reaction solution in a reaction solution BPartition between ethyl acetate and water, extract the aqueous phase with ethyl acetate (3 × 30mL), wash the combined organic extracts with brine (50mL), dry (magnesium sulfate) and concentrate in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-15% methanol in dichloromethane afforded the product which was further purified by preparative HPLC (HPLC-V-a2) to afford the desired product as a cream solid (3.7mg,0.01mmol, 13%).

HRMS-ESI(m/z)[M+H]+C₁₉H₁₅N₄O₂S₂395.0636, found 395.0659.

Example 42- {7- [ (1, 3-benzothiazol-2-yl) amino]-3, 4-dihydro-2H-1, 4-benzoxazin-4-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- (7-bromo-3, 4-dihydro-2H-1, 4-benzoxazin-4-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of benzoyl isothiocyanate (301. mu.L, 2.24mmol,1.2eq) in acetone (10mL) was added 7-bromo-3, 4-dihydro-2H-benzo [ b][1,4]Oxazine (400mg,1.87mmol,1eq) and the mixture was heated to reflux for 1 hour. The reaction was poured onto ice water and the precipitate was filtered, washed with water and dried to give a pale yellow solid. The solid was added to 1N sodium hydroxide solution (10mL) and the suspension was heated at 80 ℃ for 30 minutes, cooled to room temperature and poured onto cold 1N aqueous hydrochloric acid. The pH was adjusted to pH 8 with saturated aqueous sodium carbonate solution, the solid was collected by filtration and washed with water to give a yellow solid. A suspension of the solid and ethyl bromopyruvate (235. mu.L, 1.87mmol,1eq) in ethanol (10mL) was heated at reflux for 2 hours and then cooled to room temperature. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) purification with 0-20% ethyl acetate in isoheptaneGradient elution of solution afforded the desired product as a yellow solid (264mg,0.71mmol, 38%).

LC/MS(C₁₄H₁₃BrN₂O₃S)369[M+H]⁺；RT 1.36(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.14(d,J＝8.76Hz,1H),7.94(s,1H),7.26-7.11(m,2H),4.37-4.21(m,4H),4.07-3.99(m,2H),1.29(t,3H)。

Step B2- {7- [ (1, 3-benzothiazol-2-yl) amino]-3, 4-dihydro-2H-1, 4-benzoxazin-4-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step A (173mg,0.47mmol,1eq), 2-aminobenzothiazole (105mg,0.7mmol,1.5eq), potassium tert-butoxide (105mg,0.94mmol,2eq) and 1, 4-dioxane (5mL) and the mixture was purged with nitrogen (10min) then Brettphos (37.7mg,0.07mmol,0.15eq) and tris (dibenzylideneacetone) dipalladium (0) (42.9mg,0.05mmol,0.1eq) were added and heated at 140 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a pale yellow solid (91.4mg,0.21mmol, 45%) which was used in the next step without further purification.

LC/MS(C₂₁H₁₈N₄O₃S₂)439[M+H]⁺；RT 1.35(LCMS-V-B1)

Step C2- {7- [ (1, 3-benzothiazol-2-yl) amino]-3, 4-dihydro-2H-1, 4-benzoxazin-4-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (91.4mg,0.21mmol,1eq) in tetrahydrofuran (3mL) and methanol (1mL) was added 1N aqueous sodium hydroxide (0.42mL,0.42mmol,2eq) and the mixture was heated at 50 ℃ for 2 h. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 30mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. Purification by preparative HPLC (HPLC-V-a1) gave the desired product as a cream solid (1.5mg, 2%).

LC/MS(C₁₉H₁₄N₄O₃S₂)411[M+H]⁺；RT 1.18(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ10.51(s,1H),8.02(d,J＝8.9Hz,1H),7.81(dd,J＝7.8,1.2Hz,1H),7.69-7.59(m,3H),7.33(td,J＝7.7,1.3Hz,1H),7.24(dd,J＝8.9,2.5Hz,1H),7.16(td,J＝7.6,1.2Hz,1H),4.30(t,J＝4.4Hz,2H),4.03(t,J＝4.5Hz,2H)。

HRMS-ESI(m/z)[M+H]+C₁₉H₁₅N₄O₃S₂411.0586, found 411.0610.

Example 56- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-indol-1-yl } pyridine-2-carboxylic acid

Step A6- (5-iodo-1H-indol-1-yl) pyridine-2-carboxylic acid

To a stirring solution of 5-iodoindole (170mg,0.7mmol,1eq) in 1, 4-dioxane (5 mL)/dimethylformamide (1mL) was added sodium hydride (60% dispersion; 20.1mg,0.84mmol,1.2eq) portionwise over 20 minutes, then the mixture was stirred for 30 minutes, then ethyl 6-chloropicolinate (143mg,0.77mmol,1.1eq) was added and stirred at 70 ℃ overnight. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3x30mL), the combined organic extracts were washed with brine (3x30mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a yellow solid (38.3mg,0.11mmol, 15%).

LC/MS(C₁₄H₉IN₂O₂)365[M+H]⁺；RT 1.22(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ13.47(s,1H),8.64(d,J＝8.8Hz,1H),8.20-8.17(m,1H),8.16-8.13(m,1H),8.08-8.02(m,2H),7.93(dd,J＝7.5,0.8Hz,1H),7.59(dd,J＝1.8Hz,1H),6.78(dd,J＝3.6,0.7Hz,1H)。

Step B6- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-indol-1-yl } pyridine-2-carboxylic acid

To an oven dried microwave vial was added the product of step A (38.3mg,0.11mmol,1eq), 2-aminobenzothiazole (19mg,0.13mmol,1.2eq), sodium tert-butoxide (20.2mg,0.21mmol,2eq) and 1, 4-dioxane (2mL) and the mixture was purged with nitrogen (10min) then Brettphos (5.65mg,0.01mmol,0.1eq) and tris (dibenzylideneacetone) dipalladium (0) (9.63mg,0.01mmol,0.1eq) were added and heated under microwave irradiation at 140 ℃ for 4 hours. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. Purification by preparative HPLC (HPLC-V-a2) gave the desired product as a cream solid (0.8mg, 2%).

HRMS-ESI(m/z)[M+H]+C₂₁H₁₅N₄O₂Calculated value of S is 387.0916, found value is 387.0943.

Example 62- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-pyrrolo [2,3-b]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- { 5-bromo-1H-pyrrolo [2,3-b ] ]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of 5-bromo-1H-pyrrolo [2,3-b ] pyridine (250mg,1.27mmol,1eq) in 1, 4-dioxane (5mL) and dimethylformamide (2mL) was added sodium hydride (60% dispersion; 36.5mg,1.52mmol,1.2eq) portionwise over 20 minutes and the mixture was stirred at room temperature for 30 minutes before ethyl 2-bromo-1, 3-thiazole-4-carboxylate (449mg,1.9mmol,1.5eq) was added. The mixture was heated to reflux for 2 hours then cooled to room temperature and the precipitate formed was collected by filtration and dried in vacuo to give the desired product as a cream solid (300mg,0.85mmol, 67%).

LC/MS(C₁₃H₁₀BrN₃O₂S)352[M+H]⁺；RT 1.41(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.61(d,J＝2.2Hz,1H),8.48(d,J＝2.2Hz,1H),8.38(s,1H),8.33(d,J＝3.9Hz,1H),6.88(d,J＝3.8Hz,1H),4.34(q,J＝7.1Hz,2H),1.34(t,J＝7.1Hz,3H)。

Step B2- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-pyrrolo [2,3-b]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step A (200mg,0.57mmol,1eq), 2-aminobenzothiazole (128mg,0.85mmol,1.5eq), cesium carbonate (370mg,1.14mmol,2eq) and 1, 4-dioxane (3mL) and the mixture was purged with nitrogen (10min) then tris (dibenzylideneacetone) dipalladium (0) (52mg,0.06mmol,0.1eq) and Xantphos (64.8mg,0.12mmol,0.2eq) were added and heated at 120 ℃ for 6 hours under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 30mL), the combined organic extracts were washed with brine (30mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to give the crude desired product as a yellow gum which was used directly in the next step without further purification.

LC/MS(C₂₀H₁₅N₅O₂S₂)422[M+H]⁺；RT 1.37(LCMS-V-B1)

Step C2- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-pyrrolo [2,3-b]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (61.2mg,0.15mmol,1eq) in tetrahydrofuran (3mL) and methanol (1mL) was added 1N aqueous sodium hydroxide (0.29mL,0.29mmol,2eq) and the mixture was heated at 50 ℃ for 2 h. The reaction was concentrated in vacuo, the residue was suspended in water and acidified to pH 6 with 1N aqueous hydrochloric acid. The mixture was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 30mL), the combined organic extracts were washed with brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, c 185.5 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water afforded the desired product as a white solid (2.5mg,0.01mmol, 4%).

HRMS-ESI(m/z)[M+H]+C₁₈H₁₂N₅O₂S₂394.0432, found 394.0459.

Example 72- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-pyrrolo [2,3-c]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- { 5-chloro-1H-pyrrolo [2,3-c ]]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of 5-chloro-1H-pyrrolo [2,3-c ] pyridine (300mg,1.97mmol,1eq) in 1, 4-dioxane (5mL) and dimethylformamide (2mL) was added sodium hydride (60% dispersion; 56.6mg,2.36mmol,1.2eq) portionwise over 20 minutes and the mixture was stirred at room temperature for 30 minutes, then ethyl 2-bromo-1, 3-thiazole-4-carboxylate (696mg,2.95mmol,1.5eq) was added and heated under reflux for 2 hours. The reaction was cooled to room temperature and the precipitate formed was collected by filtration and dried in vacuo to afford the desired product as a light brown solid (518mg,1.68mmol, 86%).

LC/MS(C₁₃H₁₀ClN₃O₂S)308[M+H]⁺；RT 1.19(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ9.48(s,1H),8.39(s,1H),8.33(d,J＝3.5Hz,1H),7.85(d,J＝0.9Hz,1H),6.93(dd,J＝3.5,0.8Hz,1H),4.37(q,J＝7.1Hz,2H),1.36(t,J＝7.1Hz,3H)。

Step B2- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-pyrrolo [2,3-c]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

Add the product of step A (300mg,0.97 mm) to an oven dried microwave vialol,1eq), 2-aminobenzothiazole (220mg,1.46mmol,1.5eq), cesium carbonate (635mg,1.95mmol,2eq) and 1, 4-dioxane (7mL) and the mixture was purged with nitrogen (10min), followed by the addition of tris (dibenzylideneacetone) dipalladium (0) (89.3mg,0.1mmol,0.1eq) and Xantphos (113mg,0.19mmol,0.2eq) and heating at 130 ℃ for 8 hours under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), the combined organic extracts were washed with brine (30mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to afford the desired product as a yellow solid (17.3mg,0.04mmol, 4%).

LC/MS(C₂₀H₁₅N₅O₂S₂)422[M+H]⁺；RT 1.34(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ11.52(s,1H),9.53(t,J＝0.9Hz,1H),8.34(s,1H),8.22(d,J＝3.5Hz,1H),7.87(d,1H),7.61(d,J＝8.1Hz,1H),7.54-7.49(m,1H),7.40-7.27(m,1H),7.21-7.14(m,1H),6.94(dd,J＝3.5,0.7Hz,1H),4.40(q,J＝7.1Hz,2H),1.38(t,J＝7.1Hz,3H)。

Step C2- {5- [ (1, 3-benzothiazol-2-yl) amino]-1H-pyrrolo [2,3-c]Pyridin-1-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (17.3mg,0.04mmol,1eq) in tetrahydrofuran (3mL) and methanol (1mL) was added 1N aqueous sodium hydroxide (0.08mL,0.08mmol,2eq) and the mixture was heated at 50 ℃ for 2 h. The reaction was concentrated in vacuo, the residue was suspended in water and acidified to pH 7 with 1N aqueous hydrochloric acid. The solid was collected by filtration, washed with methanol, then ether and dried under vacuum to give the desired product as a cream solid (9.1mg,0.02mmol, 56%).

HRMS-ESI(m/z)[M+H]+C₁₈H₁₂N₅O₂S₂394.0432, found 394.0452.

Example 82- {3- [ (1, 3-benzothiazol-2-yl) amino]-7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- { 3-chloro-7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To 3-chloro-7H-pyrrolo [2, 3-c)]To a solution of pyridazine (285mg,1.86mmol,1eq) in 1, 4-dioxane (5mL) and dimethylformamide (2mL) was added sodium hydride (60% dispersion; 53.4mg,2.23mmol,1.2eq) in portions over 20 minutes, and the mixture was stirred at room temperature for 30 minutes, then ethyl 2-chlorothiazole-4-carboxylate (533mg,2.78mmol,1.5eq) was added and heated under reflux for 2 hours. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 40mL), the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a pink solid (388mg,1.26mmol, 68%).

LC/MS(C₁₂H₉ClN₄O₂S)309[M+H]⁺；RT 1.14(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.69(d,J＝3.8Hz,1H),8.46(s,1H),8.30(s,1H),6.97(d,J＝3.8Hz,1H),4.36(q,J＝7.1Hz,2H),1.35(t,J＝7.1Hz,3H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step A (388mg,1.26mmol,1eq), 2-aminobenzothiazole (283mg,1.89mmol,1.5eq), cesium carbonate (819mg,2.51mmol,2eq) and 1, 4-dioxane (10mL) and the mixture was purged with nitrogen (10min), followed by tris (dibenzylideneacetone) dipalladium (0) (115mg,0.13mmol,0.1eq) and Xantphos (145mg,0.25mmol,0.2eq) and heated at 130 ℃ for 6 hours under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 50mL), and the combined organic extracts were washed with brine (30)mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to afford the desired product as a brown solid (112mg,0.27mmol, 21%).

LC/MS(C₁₉H₁₄N₆O₂S₂)423[M+H]⁺；RT 1.29(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ11.84(s,1H),8.58(d,J＝3.9Hz,1H),8.41(s,1H),7.95(d,J＝7.8Hz,1H),7.91(s,1H),7.71-7.63(m,1H),7.40(ddd,J＝8.2,7.2,1.3Hz,1H),7.24(td,J＝7.6,1.1Hz,1H),6.99(d,J＝3.9Hz,1H),4.36(q,J＝7.1Hz,3H),1.35(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-7H-pyrrolo [2,3-c ]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (112mg,0.27mmol,1eq) in tetrahydrofuran (5mL) and methanol (1.5mL) was added 1N aqueous sodium hydroxide (0.53mL,0.53mmol,2eq) and the mixture was heated at 50 ℃ for 2 h. The reaction was concentrated in vacuo, the residue was suspended in water and the solid was collected by filtration. Purification by reverse phase automated flash chromatography (CombiFlash Rf, c 185.5 g RediSep column) eluting with a gradient of 5-95% acetonitrile in water afforded the desired product as a pale yellow solid (4.9mg,0.01mmol, 5%).

HRMS-ESI(m/z)[M+H]+C₁₇H₁₁N₆O₂S₂395.0385, found 395.0406.

Example 92- {3- [ (1, 3-benzothiazol-2-yl) amino]-7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- [3- (2-fluorophenoxy) propyl]-1, 3-thiazole-4-carboxylic acid

Step A5-bromo-2-acetylamino-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of ethyl 2-amino-5-bromothiazole-4-carboxylate (4g,15.9mmol,1eq) in dichloromethane (70mL) was added acetic anhydride (1.65mL,17.5mmol,1.1eq) and 4-dimethylaminopyridine (2.24g,18.3mmol,1.15eq) and the mixture was stirred at room temperature overnight. The reaction was cooled to room temperature, washed with water, then brine, dried (magnesium sulfate) and concentrated in vacuo. The solid formed was triturated with ether, filtered and dried in vacuo to give the desired product as an off-white solid (4.15g,14.15mmol, 89%).

LC/MS(C₈H₉BrN₂O₃S)294[M+H]⁺；RT 0.82(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ12.80(s,1H),4.28(q,J＝7.1Hz,2H),2.15(s,3H),1.30(t,J＝7.1Hz,3H)。

Step B2-acetylamino-5- (3-hydroxyprop-1-yn-1-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

Tetrakis (triphenylphosphine) palladium (0) (813mg,0.7mmol,0.05eq) was added to a solution of the product of step a (4.13g,14.1mmol,1eq), propargyl alcohol (1.64mL,28.2mmol,2eq), triethylamine (5.87mL,42.2mmol,3eq) and copper (I) iodide (0.27g,1.41mmol,0.1eq) in dimethylformamide (60mL) under a nitrogen atmosphere and the mixture was heated at 100 ℃ for 3 h. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a cream solid (3g,11.2mmol, 79%).

LC/MS(C₁₁H₁₂N₂O₄S)269[M+H]⁺；RT 0.63(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ12.80(s,1H),5.45(t,J＝6.0Hz,1H),4.37(d,J＝6.0Hz,2H),4.27(q,J＝7.1Hz,2H),2.16(s,3H),1.30(t,J＝7.1Hz,3H)。

Step C2-acetylamino-5- (3-hydroxypropyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

Ethyl acetate (30mL) and methanol (30mL) were added under nitrogen to a flask containing the product of step B (3g,11.2mmol,1eq) and hydrated platinum (IV) oxide (508mg,2.23mmol,0.2 eq). The vessel was evacuated and backfilled with nitrogen (x3), then evacuated and placed under a hydrogen atmosphere at room temperatureShake for 24 hours. The reaction was filtered through celite (10g), eluting with methanol and the solvent removed in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a brown solid (1.89g,6.94mmol, 62%).

LC/MS(C₁₁H₁₆N₂O₄S)273[M+H]⁺；RT 0.61(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ12.38(s,1H),4.54(t,J＝5.1Hz,1H),4.25(q,J＝7.1Hz,2H),3.44(q,J＝6.1Hz,2H),3.20-3.08(m,2H),2.12(s,3H),1.82-1.68(m,2H),1.29(t,J＝7.1Hz,3H)。

Step D2-amino-5- (3-hydroxypropyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step C (500mg,1.84mmol,1eq) in ethanol (15mL) was added hydrochloric acid (4M in 1, 4-dioxane; 4.59mL,4M,18.4mmol,10eq) and the mixture was heated at 60 ℃ overnight. The reaction was cooled to room temperature and then concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a beige solid (422mg,1.83mmol, 100%).

LC/MS(C₉H₁₄N₂O₃S)231[M+H]⁺；RT 0.50(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.04(br s,2H),4.26(q,J＝7.1Hz,2H),3.44(t,J＝6.3Hz,2H),3.05-2.96(m,2H),1.76-1.64(m,2H),1.29(t,J＝7.1Hz,3H)。

Step E2-bromo-5- (3-hydroxypropyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

Tert-butyl nitrate (0.26mL,2.2mmol,1.2eq) was added dropwise to a stirred solution of copper (II) bromide (491mg,2.2mmol,1.2eq) in acetonitrile (6mL) and the mixture was heated to 60 ℃ before a suspension of the product of step D (422mg,1.83mmol,1eq) in acetonitrile (8mL) was added slowly. The mixture was kept at 60 ℃ for 2 hours, then cooled to room temperature and the reaction was quenched by addition of 2N aqueous sodium hydroxide solution, then extracted with ethyl acetate. Will be provided withThe organic extracts were washed with water, brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a colorless oil (271mg,0.92mmol, 50%). LC/MS (C)₉H₁₂BrNO₃S)296[M+H]⁺；RT 0.76(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ4.59(t,J＝5.1Hz,1H),4.29(q,J＝7.1Hz,2H),3.44(td,J＝6.3,5.1Hz,2H),3.24-3.15(m,2H),1.81-1.69(m,2H),1.31(t,J＝7.1Hz,3H)。

Step F2-bromo-5- [3- (2-fluorophenoxy) propyl]-1, 3-thiazole-4-carboxylic acid ethyl ester

A solution of the product of step E (271mg,0.92mmol,1eq), 2-fluorophenol (0.12mL,1.38mmol,1.5eq) and triphenylphosphine (362mg,1.38mmol,1.5eq) in tetrahydrofuran (10mL) was cooled in an ice bath, then diisopropyl azodicarboxylate (0.27mL,1.38mmol,1.5eq) was added slowly and the mixture was stirred at 0 ℃ for 30 min and then at room temperature for 3 h. The reaction was partitioned between ethyl acetate and water, the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and eluted with a gradient of 0-60% ethyl acetate in isoheptane to give the desired product as an orange oil (302mg,0.78mmol, 85%).

LC/MS(C₁₅H₁₅BrFNO₃S)390[M+H]⁺；RT 1.23(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.26-7.07(m,3H),7.01-6.88(m,1H),4.27(q,J＝7.1Hz,2H),4.09(t,J＝6.1Hz,2H),3.39-3.29(m,2H),2.16-2.03(m,2H),1.28(t,J＝7.1Hz,3H)。

Step G2- { 3-chloro-7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } -5- [3- (2-fluorophenoxy) propyl]-1, 3-thiazole-4-carboxylic acid ethyl ester

To the stirring 3-chloro-7H-pyrrolo [2,3-c ]]To a solution of pyridazine (179mg,1.17mmol,1.5eq) in 1, 4-dioxane (10mL) and dimethylformamide (3mL) was added sodium hydride (60% dispersion; 22.4mg,0.93 mm) portionwise over 20 minutes ol,1.2eq) and the mixture is stirred for 30 minutes, then the product of step F (302mg,0.78mmol,1eq) is added, stirred at room temperature for 2 hours and at reflux overnight. The reaction was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 30mL), the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to afford the desired product as a pale yellow solid (122mg,0.27mmol, 34%).

LC/MS(C₂₁H₁₈ClFN₄O₃S)461[M+H]⁺；RT 1.41(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.63(d,J＝3.8Hz,1H),8.28(s,1H),7.26-7.07(m,3H),6.99-6.87(m,2H),4.32(q,J＝7.1Hz,2H),4.15(t,J＝6.1Hz,2H),3.48-3.37(m,2H),2.28-2.14(m,2H),1.32(t,J＝7.1Hz,3H)。

Step H2- {3- [ (1, 3-benzothiazol-2-yl) amino]-7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- [3- (2-fluorophenoxy) propyl]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a microwave vial was added the product of step G (122mg,0.27mmol,1eq), 2-aminobenzothiazole (59.7mg,0.4mmol,1.5eq), cesium carbonate (173mg,0.53mmol,2eq), tris (dibenzylideneacetone) dipalladium (0) (24.3mg,0.03mmol,0.1eq), Xantphos (15.3mg,0.03mmol,0.1eq) and 1, 4-dioxane (7.5mL) and the mixture was heated at 120 ℃ for 6 hours under microwave irradiation. The mixture was partitioned between ethyl acetate and water, the aqueous phase was extracted with ethyl acetate (3 × 40mL), washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to afford the desired product as a yellow solid (42.8mg,0.07mmol, 28%).

LC/MS(C₂₈H₂₃FN₆O₃S₂)575[M+H]⁺；RT 1.47(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ11.80(br s,1H),8.53(d,J＝3.8Hz,1H),7.95(d,J＝7.7Hz,1H),7.88(s,1H),7.69-7.63(m,1H),7.40(ddd,J＝8.2,7.3,1.3Hz,1H),7.26-7.16(m,3H),7.15-7.11(m,1H),6.98-6.91(m,2H),4.32(q,J＝7.1Hz,2H),4.17(t,J＝6.1Hz,2H),3.48-3.39(m,2H),2.28-2.17(m,2H),1.33(t,J＝7.1Hz,3H)。

Step I2- {3- [ (1, 3-benzothiazol-2-yl) amino]-7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- [3- (2-fluorophenoxy) propyl]-1, 3-thiazole-4-carboxylic acid

To a solution of the product of step H (42.8mg,0.07mmol,1eq) in 1, 4-dioxane (2mL) was added 1.25M aqueous lithium hydroxide (0.12mL,0.15mmol,2eq) and the mixture was heated at reflux for 2H. The reaction was concentrated in vacuo and purified by preparative HPLC (HPLC-V-a2) to afford the desired product as a yellow solid (2.3mg, 6%).

HRMS-ESI(m/z)[M+H]+C₂₆H₂₀FN₆O₃S₂547.1022, found 547.1010.

Example 102- {3- [ (1, 3-benzothiazol-2-yl) amino]-5H,6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

Step A3-chloro-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazine-7-carboxylic acid tert-butyl ester

To a solution of N- (but-3-yn-1-yl) -6-chloro-1, 2,4, 5-tetrazin-3-amine (381mg,2.08mmol,1eq) in tetrahydrofuran (15mL) were added di-tert-butyl dicarbonate (1.36g,6.23mmol,3eq) and 4-dimethylaminopyridine (12.7mg,0.1mmol,0.05eq) and the mixture was stirred at room temperature overnight. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to give the desired product as a red solid (89mg,0.35mmol, 17%).

LC/MS(C₁₁H₁₄ClN₃O₂)256[M+H]⁺；RT 2.06(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.62(t,J＝1.6Hz,1H),3.97(dd,J＝8.9,7.9Hz,2H),3.10(ddd,J＝9.4,7.8,1.6Hz,2H),1.51(s,9H)。

Step B3-chloro-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazines

To a solution of the product of step A (89mg,0.35mmol,1eq) in dichloromethane (3mL) was added trifluoroacetic acid (1.5mL) and the mixture was stirred at room temperature for 1 h. The reaction was concentrated in vacuo and then applied to a short column of SCX washed with methanol (5g), washed with methanol and then eluted with 1.4N methanolic ammonia to afford the desired product as a beige solid (51mg,0.33mmol, 94%).

LC/MS(C₆H₆ClN₃)156[M+H]⁺；RT 0.37(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.27(br s,1H),7.24-7.20(m,1H),3.55(td,J＝8.2,1.1Hz,2H),3.06(ddd,J＝9.7,7.8,1.7Hz,1H)。

Step C2- { 3-chloro-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step B (51mg,0.33mmol,1eq), ethyl 2-bromo-1, 3-thiazole-4-carboxylate (92.9mg,0.39mmol,1.2eq), trans-N, N' -dimethylcyclohexane-1, 2-diamine (10.3. mu.L, 0.07mmol,0.2eq), copper (I) iodide (6.24mg,0.03mmol,0.1eq), tripotassium phosphate (139mg,0.66mmol,2eq), and 1, 4-dioxane (3mL), the vessel was evacuated and flushed with nitrogen, then heated under microwave irradiation at 150 ℃ for 1 hour. The reaction was diluted with ethyl acetate, filtered through celite, then washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-3% methanol in dichloromethane afforded the crude desired product as a beige solid (19mg,0.06mmol, 19%) which was used directly in the next step without further purification.

LC/MS(C₁₂H₁₁ClN₄O₂S)311[M+H]⁺；RT 2.24(LCMS-V-C)

Step D2- {3- [ (1, 3-benzothiazol-2-yl) amino]-5H,6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step C (19mg,0.06mmol,1eq), 2-aminobenzothiazole (13.8mg,0.09mmol,1.5eq), Xantphos (7.08mg,0.01mmol,0.2eq), cesium carbonate (39.8mg,0.12mmol,2eq) and 1, 4-dioxane (3mL), the vessel was evacuated and flushed with nitrogen, then tris (dibenzylideneacetone) dipalladium (0) (5.6mg,0.01mmol,0.1eq) was added and the mixture was purged with nitrogen (10min) and then heated under microwave irradiation at 150 ℃ for 1 hour. The reaction was diluted with ethyl acetate and filtered through celite, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a brown solid (9mg,0.02mmol, 35%).

LC/MS(C₁₉H₁₆N₆O₂S₂)425[M+H]⁺；RT 2.53(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.95-7.89(m,1H),7.69-7.58(m,2H),7.41-7.35(m,1H),7.33-7.29(m,1H),7.25-7.15(m,1H),4.40-4.26(m,4H),1.33(t,3H)。

Step E2- {3- [ (1, 3-benzothiazol-2-yl) amino ]-5H,6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step D (9mg,0.02mmol,1eq) in 1, 4-dioxane (2mL) was added lithium hydroxide monohydrate (3.56mg,0.08mmol,4eq) and the mixture was heated at reflux for 6 h. The reaction was concentrated in vacuo, then dissolved in methanol and loaded onto a short column of PE-AX (5g) washed with methanol, eluted with 10:1 dichloromethane/formic acid and concentrated in vacuo. The crude material was triturated with dichloromethane, filtered and dried in vacuo to give the desired product as a beige solid (2.42mg,0.01mmol, 29%) as the formate salt.

HRMS-ESI(m/z)[M+H]+C₁₇H₁₃N₆O₂S₂397.0541, found 397.0529.

Example 112- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

Step A3-chloro-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazine

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (600mg,3.97mmol,1eq) in tetrahydrofuran (16mL) was added pent-3-yn-1-amine hydrochloride (475mg,3.97mmol,1eq) and triethylamine (553. mu.L, 3.97mmol,1eq) and the mixture was heated in a sealed tube at 110 ℃ for 8 hours. The reaction was diluted with methanol, filtered through a pad of celite and the filtrate was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, the aqueous phase was extracted with dichloromethane, the combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMCartridge silica gel) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a beige solid (96mg,0.57mmol, 14%).

LC/MS(C₇H₈ClN₃)170[M+H]⁺；RT 0.54(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.12(s,1H),3.56(td,J＝8.4,1.2Hz,2H),3.04(ddd,J＝9.2,7.9,1.3Hz,2H),2.13(d,J＝1.3Hz,3H)。

To an oven dried microwave vial was added the product of step A (96mg,0.57mmol,1eq), ethyl 2-bromo-1, 3-thiazole-4-carboxylate (187mg,0.79mmol,1.4eq), trans-N, N' -dimethylcyclohexane-1, 2-diamine (17.9 μ L,0.11mmol,0.2eq), copper (I) iodide (10.8mg,0.06mmol,0.1eq), tripotassium phosphate (240mg,1.13mmol,2eq), and 1, 4-dioxane (8mL), the vessel was evacuated and flushed with nitrogen, then heated under microwave irradiation at 150 ℃ for 1 hour. The reaction was diluted with ethyl acetate, filtered through celite, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) purification with 0-100% ethyl acetateGradient elution of the ester in isoheptane afforded the desired product as a beige solid (18mg,0.06mmol, 10%).

LC/MS(C₁₃H₁₃ClN₄O₂S)325[M+H]⁺；RT 2.32(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.11(s,1H),4.41(dd,J＝8.8,7.6Hz,2H),4.30(q,J＝7.1Hz,2H),3.34-3.27(m,2H),2.29(d,J＝1.2Hz,3H),1.31(t,J＝7.1Hz,3H)。

To an oven dried microwave vial was added the product of step B (27mg,0.08mmol,1eq), 2-aminobenzothiazole (18.7mg,0.12mmol,1.5eq), Xantphos (9.62mg,0.02mmol,0.2eq), cesium carbonate (54.2mg,0.17mmol,2eq) and 1, 4-dioxane (4mL), the vessel was evacuated and flushed with nitrogen, then tris (dibenzylideneacetone) dipalladium (0) (7.61mg,0.01mmol,0.1eq) was added and the mixture was purged with nitrogen (10min) and then heated under microwave irradiation at 150 ℃ for 1 hour. The reaction was diluted with ethyl acetate and filtered through celite, then washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a yellow solid (15mg,0.03mmol, 41%).

LC/MS(C₂₀H₁₈N₆O₂S₂)439[M+H]⁺；RT 2.67(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.06(s,1H),7.88(s,1H),7.53(br s,1H),7.38(t,J＝7.5Hz,1H),7.20(t,J＝7.6Hz,1H),4.38(t,J＝8.0Hz,2H),4.31(q,J＝7.1Hz,2H),3.32-3.21(m,2H),2.33(s,3H),1.32(t,J＝7.1Hz,3H)。

Step D2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step C (15mg,0.03mmol,1eq) in 1, 4-dioxane (2mL) was added lithium hydroxide monohydrate (5.74mg,0.14mmol,4eq) and the mixture was heated at reflux for 3 h. The reaction was concentrated in vacuo, dissolved in methanol, applied to a short column of PE-AX (5g) washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane, filtered and dried in vacuo to give the desired product as a cream solid (9.03mg,0.02mmol, 64%).

HRMS-ESI(m/z)[M+H]+C₁₈H₁₅N₆O₂S₂411.0698, found 411.0701.

Example 122- {3- [ (1, 3-benzothiazol-2-yl) amino]-5H,6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

Step APent-4-yn-1-yl methanesulfonate

To a solution of 4-pentyn-1-ol (3.32mL,35.7mmol,1eq) in dichloromethane (60mL) was added triethylamine (6.45mL,46.4mmol,1.3eq) and the mixture was cooled to 0 ℃, then methanesulfonyl chloride (3.31mL,42.8mmol,1.2eq) was added dropwise and stirred at room temperature overnight. The reaction was partitioned between dichloromethane and water and the organic phase was washed successively with saturated sodium bicarbonate and brine, dried (magnesium sulfate) and concentrated in vacuo to afford the desired product as an amber oil (5.8g,35.8mmol, 100%).

¹H NMR(400MHz,DMSO-d6)δ4.26(t,J＝6.2Hz,2H),3.19(s,3H),2.88(t,J＝2.7Hz,1H),2.29(td,J＝7.1,2.7Hz,2H),1.91-1.80(m,2H)。

Step B5-azidopent-1-yne

To a solution of the product of step A (5.8g,35.8mmol,1eq) in dimethylformamide (30mL) was added sodium azide (5.81g,89.4mmol,2.5eq) and the mixture was heated at 70 ℃ for 3 hours. The reaction was diluted with water, the aqueous phase was extracted with ether (× 3), the combined organic phases were dried (magnesium sulphate) and concentrated in vacuo to give the desired product as a yellow oil (5.65g,51.8mmol, > 100%).

¹H NMR(400MHz,DMSO-d6)δ3.42(t,J＝6.7Hz,2H),2.85(t,J＝2.7Hz,1H),2.25(td,J＝7.0,2.7Hz,2H),1.75-1.64(m,2H)。

Step CPent-4-yne-1-amines

To a solution of the product of step B (3.9g,35.7mmol,1eq) cooled to 0 ℃ in diethyl ether (40mL) was added triphenylphosphine (14.1g,53.6mmol,1.5eq) and the reaction was stirred at 0 ℃ for 6 h. The reaction was quenched by the addition of water (5mL) and stirred at room temperature overnight. The mixture was poured onto 4N aqueous hydrochloric acid (300mL) and extracted with ether (× 3). The aqueous phase was basified with sodium hydroxide added portionwise and further extracted with diethyl ether (× 2). The combined organic extracts were dried (magnesium sulfate) and concentrated in vacuo to afford the desired product as a yellow oil (1.51g,18.16mmol, 51%).

¹H NMR(400MHz,DMSO-d6)δ2.73(t,1H),2.58(t,J＝6.7Hz,2H),2.19(td,J＝7.2,2.7Hz,2H),1.55-1.44(m,2H)。

Step D2- [ (pent-4-yn-1-yl) amino group]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of ethyl 2-bromo-1, 3-thiazole-4-carboxylate (750mg,3.18mmol,1eq) in acetonitrile (15mL) was added the product of step C (396mg,4.77mmol,1.5eq) and triethylamine (0.66mL,4.77mmol,1.5eq) and the mixture was heated at 150 ℃ for 10 h under microwave irradiation. The reaction was partitioned between ethyl acetate and brine and the organic phase was dried (MgSO ₄) And concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a colorless solid (263mg,1.1mmol, 35%).

LC/MS(C₁₁H₁₄N₂O₂S)239[M+H]⁺；RT 2.20(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.84(t,J＝5.4Hz,1H),7.51(s,1H),4.22(q,J＝7.1Hz,2H),3.28(td,J＝6.9,5.4Hz,2H),2.82(t,J＝2.6Hz,1H),2.25(td,J＝7.1,2.7Hz,2H),1.73(p,J＝7.0Hz,2H),1.26(t,J＝7.1Hz,3H)。

Step E2- { 3-chloro-5H, 6H,7H, 8H-pyrido [2,3-c ]]Pyridazin-8-yl } -1, 3-thia-inesAzole-4-carboxylic acid ethyl ester

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (103mg,0.68mmol,1eq) in tetrahydrofuran (12mL) was added the product of step D (163mg,0.68mmol,1eq) and the mixture was heated at 90 ℃ overnight. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to afford the desired product as an off white solid (141mg,0.43mmol, 64%).

LC/MS(C₁₃H₁₃ClN₄O₂S)325[M+H]⁺；RT 2.42(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.08(s,1H),7.77-7.71(m,1H),4.40-4.33(m,2H),4.30(q,2H),2.94(t,J＝6.1Hz,2H),2.11-2.00(m,2H),1.32(t,J＝7.1Hz,3H)。

Step F2- {3- [ (1, 3-benzothiazol-2-yl) amino]-5H,6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step E (141mg,0.43mmol,1eq), 2-aminobenzothiazole (97.8mg,0.65mmol,1.5eq), Xantphos (50.2mg,0.09mmol,0.2eq), cesium carbonate (283mg,0.87mmol,2eq) and 1, 4-dioxane (15mL), the vessel was evacuated and flushed with nitrogen, then tris (dibenzylideneacetone) dipalladium (0) (39.8mg,0.04mmol,0.1eq) was added and the mixture was purged with nitrogen (10min) and then heated under microwave irradiation at 150 ℃ for 2 hours. The reaction was diluted with ethyl acetate and filtered through celite, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to afford the desired product as a yellow solid (29mg,0.07mmol, 15%).

LC/MS(C₂₀H₁₈N₆O₂S₂)439[M+H]⁺；RT 2.64(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.67(s,1H),8.02(s,1H),7.98(d,1H),7.66(d,J＝7.9Hz,1H),7.42(dt,J＝15.0,7.2Hz,1H),7.35(s,1H),7.23(t,J＝7.5Hz,1H),4.41-4.24(m,4H),2.96(t,2H),2.12-2.02(m,2H),1.32(t,J＝7.1Hz,3H)。

Step G2- {3- [ (1, 3-benzothiazol-2-yl) amino]-5H,6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step F (29mg,0.07mmol,1eq) in 1, 4-dioxane (6mL) was added lithium hydroxide monohydrate (13.9mg,0.33mmol,5eq) and the mixture was heated at reflux for 5 h. The reaction was concentrated in vacuo, dissolved in methanol and then applied to a short column of PE-AX (5g) washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane, filtered and dried in vacuo to give the desired product as a cream solid (9.86mg,0.02mmol, 36%) as formate.

HRMS-ESI(m/z)[M+H]+C₁₈H₁₅N₆O₂S₂411.0698, found 411.0722.

Example 135- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

Step A5- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy ]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven-dried microwave vial were added the product of preparation 6a (98mg,0.15mmol,1eq), the product of preparation 5a (64.7mg,0.18mmol,1.2eq), potassium carbonate (62.7mg,0.45mmol,3eq), [1,1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride (11.1mg,0.02mmol,0.1eq), tetrahydrofuran (3mL) and water (1mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the organic layer was washed with brine, dried (magnesium sulfate) and vacuumAnd (4) concentrating. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a cream solid (57mg,0.07mmol, 47%).

¹H NMR(400MHz,DMSO-d6)δ7.78(d,1H),7.56(s,1H),7.48-7.38(m,2H),7.27-7.20(m,1H),5.85(s,2H),4.37(t,J＝8.1Hz,2H),4.17(q,J＝7.1Hz,2H),3.79(s,2H),3.76-3.67(m,2H),3.45-3.36(m,2H),2.34(s,3H),2.23(s,3H),2.02-1.90(m,3H),1.73-1.52(m,12H),1.16(t,3H),0.96-0.87(m,2H),-0.11(s,9H)。

Step B5- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a cooled solution of the product of step A (57mg,0.07mmol,1eq) in dichloromethane (6mL) was added trifluoroacetic acid (0.6mL) and after 10 minutes the mixture was warmed to room temperature and stirred overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a yellow solid (17mg,0.03mmol, 36%).

LC/MS(C₃₅H₃₈N₈O₂S₂)667[M+H]⁺；RT 1.55(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.85(d,J＝7.6Hz,1H),7.62-7.44(m,2H),7.42-7.31(m,1H),7.20(t,J＝7.6Hz,1H),4.37(t,J＝8.1Hz,2H),4.18(q,J＝7.1Hz,2H),3.80(s,2H),3.34-3.24(m,2H),2.34(d,J＝3.4Hz,3H),2.24(s,3H),2.02-1.93(m,3H),1.74-1.51(m,12H),1.18(t,3H)。

Step C5- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (17mg,0.03mmol,1eq) in 1, 4-dioxane (6mL) was added lithium hydroxide monohydrate (10.7mg,0.25mmol,10eq) and the mixture was heated at reflux for 5 h. The reaction was concentrated in vacuo, dissolved in methanol, applied to a short column of PE-AX (5g) washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with ether and acetonitrile, filtered and dried in vacuo to give the desired product as a beige solid (2.4mg,3.7 μmol, 15%).

HRMS-ESI(m/z)[M+H]+C₃₃H₃₅N₈O₂S₂639.2324, found 639.2310.

Example 142- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (1- { [1- (3-methoxypropyl) cyclooctyl radical]Methyl } -5-methyl-1H-pyrazol-4-yl) -1, 3-thiazole-4-carboxylic acid

Step A5- (1- { [1- (3-methoxypropyl) cyclooctyl group]Methyl } -5-methyl-1H-pyrazol-4-yl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy ]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of preparation 6a (34mg,0.05mmol,1eq), the product of preparation 5b (25.5mg,0.06mmol,1.2eq), potassium carbonate (21.8mg,0.16mmol,3eq), [1,1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride (3.84mg,0.01mmol,0.1eq), tetrahydrofuran (3mL) and water (1mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between ethyl acetate and water, and the organic layer was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a white solid (29mg,0.03mmol, 65%).

LC/MS(C₄₃H₆₀N₈O₄SiS₂)845[M+H]⁺；RT 1.79(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.77(d,1H),7.58(s,1H),7.49-7.38(m,2H),7.27-7.19(m,1H),5.85(s,2H),4.37(t,J＝8.2Hz,2H),4.17(q,J＝7.1Hz,2H),3.85(s,2H),3.77-3.66(m,2H),3.45-3.34(m,2H),3.31-3.26(m,4H),3.23(s,3H),2.33(s,3H),2.22(s,3H),1.74-1.48(m,8H),1.47-1.20(m,8H),1.18(t,3H),0.96-0.87(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (1- { [1- (3-methoxypropyl) cyclooctyl radical]Methyl } -5-methyl-1H-pyrazol-4-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a cooled solution of the product of step A (29mg,0.03mmol,1eq) in dichloromethane (5mL) was added trifluoroacetic acid (0.9mL) and after 10 minutes the mixture was warmed to room temperature and stirred overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, dried (PTFE phase separator) and concentrated in vacuo to afford the desired product as a yellow solid (13mg,0.02mmol, 54%).

LC/MS(C₃₇H₄₆N₈O₃S₂)715[M+H]⁺；RT 1.59(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.86(s,1H),7.59(br s+s,2H),7.37(t,1H),7.20(t,J＝7.6Hz,1H),4.37(t,J＝8.1Hz,2H),4.19(q,J＝7.0Hz,2H),3.87(s,2H),3.34-3.26(m,6H),3.25(s,3H),2.34(s,3H),2.23(s,3H),1.73-1.49(m,8H),1.48-1.21(m,8H),1.18(t,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (1- { [1- (3-methoxypropyl) cyclooctyl radical]Methyl } -5-methyl-1H-pyrazol-4-yl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (13mg,0.02mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (11.5mg,0.27mmol,15eq) and the mixture was heated at reflux for 5 h. The reaction was concentrated in vacuo, the residue triturated with water, filtered and dried in vacuo to afford the desired product as a yellow solid (5.64mg,0.01mmol, 45%) as the lithium salt.

HRMS-ESI(m/z)[M+H]+C₃₅H₄₃N₈O₃S₂Calculated value of (2) 687.2900, found value of 687.2932

Example 152- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of preparation 3f (24mg,0.05mmol,1eq) in 1, 4-dioxane (6mL) was added lithium hydroxide monohydrate (33.4mg,0.8mmol,15eq) and the mixture was heated at reflux for 7 h. The reaction was concentrated in vacuo and then dissolved in methanol, applied to a short column of PE-AX (5g) wetted with methanol, washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane, filtered and dried in vacuo to give the desired product as a beige solid (13.5mg,0.03mmol, 60%) as the formate salt.

HRMS-ESI(m/z)[M+H]+C₁₉H₁₇N₆O₂S₂425.0854, found 425.0845.

Example 162- {3- [ (1, 3-benzothiazol-2-yl) amino]-6-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- [ (pent-4-yn-2-yl) amino group]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of ethyl 2-bromo-1, 3-thiazole-4-carboxylate (1.87g,7.93mmol,1eq) in acetonitrile (18mL) was added pent-4-yn-2-amine (989mg,11.9mmol,1.5eq) and triethylamine (1.66mL,11.9mmol,1.5eq) and the mixture was sealed in a sealing testThe tube was heated at 170 ℃ overnight. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a yellow oil (555mg,2.33mmol, 29%).

LC/MS(C₁₁H₁₄N₂O₂S)239[M+H]⁺；RT 2.21(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.85(d,J＝7.4Hz,1H),7.51(s,1H),4.27(q,2H),3.91-3.79(m,1H),2.89(t,J＝2.6Hz,1H),2.51-2.45(m,1H),2.44-2.41(m,1H),1.30-1.21(m,6H)。

Step B2- { 3-chloro-6-methyl-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (352mg,2.33mmol,1eq) in tetrahydrofuran (15mL) was added the product of step A (555mg,2.33mmol,1eq) and the mixture was heated at reflux overnight. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to afford the desired product as a red solid (124mg,0.38mmol, 16%).

LC/MS(C₁₃H₁₃ClN₄O₂S)325[M+H]⁺；RT 2.39(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.12(s,1H),7.71(t,J＝1.6Hz,1H),5.11-4.97(m,1H),4.31(q,J＝7.1,1.4Hz,2H),3.65-3.53(m,1H),3.00-2.88(m,1H),1.50(d,J＝6.3Hz,3H),1.31(t,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-6-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step B (124mg,0.38mmol,1eq), 2-aminobenzothiazole (86mg,0.57mmol,1.5eq), Xantphos (44.2mg,0.08mmol,0.2eq), cesium carbonate (249mg,0.76mmol,2eq), 1, 4-dioxane (4mL) and tris (dibenzylideneacetone) dipalladium (0) (35mg,0.04 mmol)0.1eq) and the mixture was purged with nitrogen (10min) and then heated under microwave irradiation at 150 ℃ for 2 hours. The reaction was diluted with ethyl acetate and filtered through celite, then washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give a solid which was triturated with ether, filtered and dried in vacuo to give the desired product as a beige solid (37mg,0.08mmol, 22%).

LC/MS(C₂₀H₁₈N₆O₂S₂)439[M+H]⁺；RT 2.62(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.69(s,1H),8.07(s,1H),7.95(d,J＝7.9Hz,1H),7.66(d,J＝8.1Hz,1H),7.44-7.37(m,1H),7.36(s,1H),7.23(td,J＝7.6,1.1Hz,1H),5.07-4.95(m,1H),4.31(q,2H),3.65-3.52(m,1H),3.03-2.93(m,1H),1.49(d,J＝6.3Hz,3H),1.32(t,J＝7.1Hz,3H)。

Step D2- {3- [ (1, 3-benzothiazol-2-yl) amino]-6-methyl-5H, 6H, 7H-pyrrolo [2,3-c ]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To the product of step C (37mg,0.08mmol,1eq) in 1, 4-dioxane (8mL) was added lithium hydroxide monohydrate (53.1mg,1.27mmol,15eq) and the mixture heated to reflux for 7 h. The reaction was concentrated in vacuo, dissolved in methanol, then applied to a short column of PE-AX (10g) wetted with methanol, washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane, filtered and dried in vacuo to give the desired product as an off-white solid (24.8mg,0.06mmol, 72%) as the formate salt.

HRMS-ESI(m/z)[M+H]+C₁₈H₁₅N₆O₂S₂411.0698, found 411.0695.

Example 172- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (1- { [1- (3-methoxypropyl) cyclohexyl]Methyl } -5-methyl-1H-pyrazol-4-yl) -1, 3-thiazole-4-carboxylic acid

Step A5- (1- { [1- (3-methoxypropyl) cyclohexyl]Methyl } -5-methyl-1H-pyrazol-4-yl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of preparation 6a (37mg,0.06mmol,1eq), the product of preparation 5c (25.8mg,0.07mmol,1.2eq), potassium carbonate (23.7mg,0.17mmol,3eq), [1,1' -bis (diphenylphosphino) ferrocene ]Palladium (II) dichloride (4.18mg,0.01mmol,0.1eq), tetrahydrofuran (3mL) and water (1mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to afford the desired product as a white solid (22mg,0.03mmol, 47%).

LC/MS(C₄₁H₅₆N₈O₄SiS₂)818[M+H]⁺；RT 3.45(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.79-7.74(m,1H),7.58(s,1H),7.48-7.38(m,2H),7.27-7.20(m,1H),5.85(s,2H),4.37(t,J＝8.1Hz,2H),4.17(q,J＝7.1Hz,2H),3.93(s,2H),3.76-3.66(m,2H),3.44-3.36(m,2H),3.34-3.25(m,2H),3.23(s,3H),2.33(s,3H),2.23(s,3H),1.60-1.27(m,14H),1.18(t,J＝7.1Hz,3H),0.98-0.85(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (1- { [1- (3-methoxypropyl) cyclohexyl]Methyl } -5-methyl-1H-pyrazol-4-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of cooling step A (22mg,0mol,1eq) in dichloromethane (5mL) was added trifluoroacetic acid (1.5mL) and after 10 minutes the mixture was warmed to room temperatureAnd stirred overnight. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to afford the desired product as a yellow solid (10mg,0.01mmol, 54%).

LC/MS(C₃₅H₄₂N₈O₃S₂)688[M+H]⁺；RT 3.02(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.91-7.79(m,1H),7.59(br s+s,2H),7.38(t,J＝7.7Hz,1H),7.25-7.13(m,1H),4.38(t,J＝8.2Hz,2H),4.19(q,J＝7.0Hz,2H),3.95(s,2H),3.34-3.27(m,4H),3.25(s,3H),2.34(s,3H),2.24(s,3H),1.60-1.45(m,6H),1.44-1.29(m,6H),1.28-1.22(m,2H),1.18(t,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (1- { [1- (3-methoxypropyl) cyclohexyl)]Methyl } -5-methyl-1H-pyrazol-4-yl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (10mg,0.01mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (12.2mg,0.29mmol,20eq) and the mixture was heated at reflux for 5 h. The reaction was concentrated in vacuo, triturated with water, filtered and dried in vacuo to afford the desired product as a yellow solid (5.71mg,0.01mmol, 60%).

HRMS-ESI(m/z)[M+H]+C₃₃H₃₉N₈O₃S₂659.2587, found 659.2577.

Example 182- { 4-methyl-3- [ (1, 3-thiazol-2-yl) amino group]-5H,6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- { 4-methyl-3- [ (1, 3-thiazol-2-yl) amino group]-5H,6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of preparation 6a, step B (100mg,0.31mmol,1eq), 2-aminothiazole (46.3mg,0.46mmol,1.5eq), Xantphos (35.6mg,0.06mmol,0.2eq), cesium carbonate (201mg,0.62mmol,2eq), 1, 4-dioxane (4mL) and tris (dibenzylideneacetone) dipalladium (0) (28.2mg,0.03mmol,0.1eq) and the mixture was purged with nitrogen (10min) and then heated at 150 ℃ for 1 hour under microwave irradiation. The reaction was diluted with ethyl acetate, filtered through celite, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded a solid which was triturated with acetonitrile, filtered and dried in vacuo to afford the desired product as a yellow solid (23mg,0.06mmol, 19%).

LC/MS(C₁₆H₁₆N₆O₂S₂)389[M+H]⁺；RT 2.29(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ10.57(br s,1H),8.04(s,1H),7.44(br s,1H),7.06(br s,1H),4.40-4.32(m,2H),4.29(q,2H),3.30-3.22(m,2H),2.31(s,3H),1.32(t,3H)。

Step B2- { 4-methyl-3- [ (1, 3-thiazol-2-yl) amino group]-5H,6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step A (23mg,0.06mmol,1eq) in 1, 4-dioxane (6mL) was added lithium hydroxide monohydrate (37.3mg,0.89mmol,15eq) and the mixture was heated at reflux for 5 h. The reaction was concentrated in vacuo, dissolved in methanol, applied to a short column of PE-AX (5g) wetted with methanol, washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane and methanol, filtered and dried in vacuo to give the desired product as a beige solid (8.84mg,0.02mmol, 41%).

HRMS-ESI(m/z)[M+H]+C₁₄H₁₃N₆O₂S₂361.0541, found 361.0531.

Example 192- {3- [ (4, 5-dimethyl-1, 3-thiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thia-inesAzole-4-carboxylic acid

Step A2- {3- [ (4, 5-dimethyl-1, 3-thiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of preparation 6a, step B (100mg,0.31mmol,1eq), 4, 5-dimethyl-1, 3-thiazol-2-amine (59.2mg,0.46mmol,1.5eq), Xantphos (35.6mg,0.06mmol,0.2eq), cesium carbonate (201mg,0.62mmol,2eq), 1, 4-dioxane (3mL) and tris (dibenzylideneacetone) dipalladium (0) (28.2mg,0.03mmol,0.1eq) and the mixture was purged with nitrogen (10min) and then heated at 150 ℃ for 1 hour under microwave irradiation. The reaction was diluted with ethyl acetate and filtered through celite, then washed with brine, dried (magnesium sulfate) and concentrated in vacuo. The residue was triturated with methanol, filtered and dried in vacuo to give the desired product as a yellow solid (64mg,0.15mmol, 50%).

LC/MS(C₁₈H₂₀N₆O₂S₂)417[M+H]⁺；RT 2.42(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.03(s,1H),4.39-4.21(m,4H),3.25(t,J＝8.0Hz,2H),2.27(s,3H),2.23(s,3H),2.16(s,3H),1.31(t,J＝7.1Hz,3H)。

Step B2- {3- [ (4, 5-dimethyl-1, 3-thiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step A (64mg,0.15mmol,1eq) in 1, 4-dioxane (15mL) was added lithium hydroxide monohydrate (96.7mg,2.3mmol,15eq) and the mixture was heated at reflux for 5 h. The reaction was concentrated in vacuo, dissolved in methanol and then applied to a short column of PE-AX (10g) wetted with methanol, washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with methanol, filtered and dried in vacuo to give the desired product as a beige solid (17.9mg,0.05mmol, 30%).

HRMS-ESI(m/z)[M+H]+C₁₆H₁₇N₆O₂S₂389.0854, found 389.0847.

Example 206- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } pyridine-2-carboxylic acid

Step A6- [ (pent-3-yn-1-yl) amino group]Pyridine-2-carboxylic acid tert-butyl ester

To a solution of tert-butyl 6-fluoropyridine-2-carboxylate (219mg,1.11mmol,1eq) in dimethylacetamide (5mL) was added pent-3-yn-1-amine hydrochloride (133mg,1.11mmol,1eq) and N, N-diisopropylethylamine (0.39mL,2.22mmol,2eq) and the mixture was heated in a sealed tube at 120 ℃ overnight. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a clear oil (48mg,0.18mmol, 17%).

LC/MS(C₁₅H₂₀N₂O₂)261[M+H]⁺；RT 2.42(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.49(dd,J＝8.4,7.2Hz,1H),7.11(dd,J＝7.3,0.8Hz,1H),6.89(t,J＝5.7Hz,1H),6.66(dd,J＝8.5,0.8Hz,1H),3.45-3.29(m,2H),2.44-2.35(m,2H),1.75(t,J＝2.6Hz,3H),1.53(s,9H)。

Step B6- { 3-chloro-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } pyridine-2-carboxylic acid tert-butyl ester

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (27.8mg,0.18mmol,1eq) in tetrahydrofuran (3mL) was added the product of step a (48mg,0.18mmol,1eq) and the mixture was heated under microwave irradiation at 110 ℃ for 1 hour. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMSilica gel cartridge), gradient eluting with 0-30% ethyl acetate in isoheptane solution to obtainDesired product was obtained as a pink solid (12mg,0.03mmol, 19%).

LC/MS(C₁₇H₁₉ClN₄O₂)347[M+H]⁺；RT 2.67(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.84(dd,J＝8.6,0.8Hz,1H),8.01(dd,J＝8.6,7.4Hz,1H),7.65(dd,J＝7.4,0.8Hz,1H),4.36(dd,J＝8.9,7.8Hz,2H),3.22(t,J＝8.3Hz,2H),2.27(s,3H),1.57(s,9H)。

Step C6- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } pyridine-2-carboxylic acid tert-butyl ester

To an oven dried microwave vial was added the product of step B (26mg,0.07mmol,1eq), 2-aminothiazole (16.9mg,0.11mmol,1.5eq), Xantphos (8.68mg,0.01mmol,0.2eq), cesium carbonate (48.9mg,0.15mmol,2eq), 1, 4-dioxane (4mL) and tris (dibenzylideneacetone) dipalladium (0) (6.87mg,0.01mmol,0.1eq) and the mixture was purged with nitrogen (10min) and then heated at 150 ℃ for 1 hour under microwave irradiation. The reaction was diluted with ethyl acetate, filtered through celite, washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give a solid which was triturated with ether, filtered and dried in vacuo to give the desired product as a yellow solid (7mg,0.02mmol, 20%).

LC/MS(C₂₄H₂₄N₆O₂S)461[M+H]⁺；RT 2.69(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.98(d,J＝8.7Hz,1H),8.01(t,J＝8.0Hz,1H),7.88(s,1H),7.68-7.57(m,2H),7.38(t,J＝7.5Hz,1H),7.28-7.14(m,1H),4.32(t,J＝8.2Hz,2H),3.20(t,J＝8.1Hz,2H),2.33(s,3H),1.58(s,8H)。

Step D6- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c ]Pyridazin-7-yl } pyridine-2-carboxylic acid

To a solution of the product of step C (7mg,0.02mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (9.57mg,0.23mmol,15eq) and the mixture was heated at reflux for 6 h. The reaction was concentrated in vacuo and then dissolved in methanol, applied to a short column of PE-AX (5g) wetted with methanol, washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane and methanol, filtered and dried in vacuo to give the desired product as a yellow solid (2.0mg, 32%).

HRMS-ESI(m/z)[M+H]+C₂₀H₁₇N₆O₂Calculated value of S405.1134, found value 405.1122.

Example 216- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } pyridine-2-carboxylic acid

Step A6- { [3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl]Amino } pyridine-2-carboxylic acid ethyl ester

To a solution of the product of preparation 2e (500mg,2.28mmol,1eq) and ethyl 6-aminopyralid (455mg,2.74mmol,1.2eq) in methanol (18mL) and acetic acid (6mL) was added sodium triacetoxyborohydride (968mg,4.56mmol,2eq) and the mixture was stirred at room temperature for 16 h. The reaction was quenched by the addition of 1N aqueous sodium hydroxide (50mL), extracted with ethyl acetate (3 × 50mL) and the combined organic extracts were washed successively with saturated aqueous sodium bicarbonate and brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to give the desired product as a white solid (400mg,1.08mmol, 47%).

LC/MS(C₁₆H₁₈Cl₂N₄O₂)369[M+H]⁺；RT 1.19(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.52(dd,J＝8.4,7.2Hz,1H),7.18(dd,1H),6.99(t,J＝5.7Hz,1H),6.69(dd,J＝8.5,0.8Hz,1H),4.27(q,J＝7.1Hz,2H),3.42-3.35(m,2H),2.92-2.82(m,2H),2.41(s,3H),1.87-1.74(m,2H),1.29(t,J＝7.1Hz,3H)。

Step B6- { 3-chloro-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]]Pyridazin-8-yl } pyridine-2-carboxylic acid methyl ester

To a solution of the product of step a (170mg,0.46mmol,1eq) in α, α, α -trifluorotoluene (4mL) was added cesium carbonate (300mg,0.92mmol,2eq) and the mixture was heated in a sealed tube at 160 ℃ for 5 days. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a cream solid (74mg,0.23mmol, 50%).

LC/MS(C₁₅H₁₅ClN₄O₂)319[M+H]⁺；RT 1.10(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.97-7.84(m,2H),7.75(dd,J＝7.1,1.2Hz,1H),4.05-3.96(m,2H),3.89(s,3H),2.92-2.81(m,2H),2.31(s,3H),2.05-1.92(m,2H)。

Step C6- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } pyridine-2-carboxylic acid methyl ester

To a solution of the product of step B (74mg,0.23mmol,1eq), 2-aminobenzothiazole (52.3mg,0.35mmol,1.5eq) and N, N-diisopropylethylamine (0.12mL,0.7mmol,3eq) in 1, 4-dioxane (5mL) was added Xantphos (13.4mg,0.02mmol,0.1eq) and tris (dibenzylideneacetone) dipalladium (0) (10.6mg,0.01mmol,0.05eq) and the mixture was heated in a sealed tube at 150 ℃ for 20 h. The reaction was cooled to room temperature, then partitioned between ethyl acetate (20mL) and brine (25mL), the organic phase dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a yellow solid (20mg,0.05mmol, 20%).

LC/MS(C₂₂H₂₀N₆O₂S)433[M+H]⁺；RT 1.15(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.03(dd,J＝8.5,0.9Hz,1H),7.87(dd,J＝8.5,7.3Hz,1H),7.83(br s,1H),7.68(d,J＝7.3Hz,1H),7.53(br s,1H),7.36(t,J＝7.6Hz,1H),7.17(t,J＝7.6Hz,1H),4.11-3.99(m,2H),3.90(s,3H),2.86(t,J＝6.5Hz,2H),2.33(s,3H),2.05-1.94(m,2H)。

Step D6- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } pyridine-2-carboxylic acid

To a solution of the product of step C (15mg,0.03mmol,1eq) in 1, 4-dioxane (2mL) was added lithium hydroxide monohydrate (2.91mg,0.07mmol,2eq) and the mixture was heated at reflux for 1 h. The reaction was cooled to room temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, c184.3g RediSep column) with a gradient of 5-95% acetonitrile in water afforded the desired product as a cream solid (10mg,0.02mmol, 69%).

HRMS-ESI(m/z)[M+H]+C₂₁H₁₉N₆O₂Calculated value of S is 419.1290, found value is 419.1287.

Example 222- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- [ (1E) -3- [ (tert-butyldimethylsilyl) oxy group]Prop-1-en-1-yl]-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene ]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried sealed flask was added the product of preparation 6a (3g,4.63mmol,1eq), (E) -3- (tert-butyldimethylsilyloxy) propen-1-yl-boronic acid pinacol ester (1.82mL,5.56mmol,1.2eq), potassium carbonate (1.92g,13.9mmol,3eq), [1,1' -bis (diphenylphosphine) ferrocene]Palladium (II) dichloride (339mg,0.46mmol,0.1eq), tetrahydrofuran (150mL) and water (50mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ for 1.5 h. The reaction solution was cooled to room temperatureThen partitioned between ethyl acetate and water and the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to give the desired product as a cream solid (1.86g,2.52mmol, 54%).

LC/MS(C₃₅H₅₀N₆O₄Si₂S₂)739[M+H]⁺；RT 3.69(Shortneg2)

¹H NMR(400MHz,DMSO-d6)δ7.80(d,J＝7.6Hz,1H),7.55-7.38(m,3H),7.30-7.20(m,1H),6.30(dt,J＝15.9,4.3Hz,1H),5.85(s,2H),4.41-4.26(m,4H),3.77-3.67(m,2H),3.45-3.20(m,4H),2.32(s,3H),1.32(t,J＝7.1Hz,3H),0.93(s,9H),0.92-0.86(m,2H),0.11(s,6H),-0.11(s,9H)。

Step B5- {3- [ (tert-butyldimethylsilyl) oxy group]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (900mg,1.22mmol,1eq) in ethyl acetate (600mL) under nitrogen was added a catalytic amount of platinum (IV) oxide. The mixture was evacuated and backfilled with nitrogen (x3), then evacuated and backfilled with hydrogen and shaken at room temperature under a hydrogen atmosphere for 3 days. The reaction was filtered through celite, eluted with ethyl acetate and evaporated under reduced pressure to give the desired product as a beige solid (950mg,1.28mmol, > 100%%).

LC/MS(C₃₅H₅₂N₆O₄Si₂S₂)741[M+H]⁺；RT 1.88(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.77(d,J＝7.6Hz,1H),7.50-7.39(m,2H),7.28-7.18(m,1H),5.85(s,2H),4.37-4.22(m,4H),3.77-3.62(m,4H),3.31-3.14(m,4H),2.32(s,3H),1.93-1.80(m,2H),1.31(t,J＝7.1Hz,3H),0.96-0.80(m,11H),0.06(s,6H),-0.11(s,9H)。

Step C5- (3-hydroxypropyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethyl esterOxy radical]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step B (950mg,1.28mmol,1eq) in 1, 4-dioxane (150mL) was added hydrochloric acid (4M in dioxane; 50mL,200mmol,156eq) and the mixture was stirred at room temperature for 1 h. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-80% ethyl acetate in isoheptane to afford the desired product as an off white solid (577mg,0.92mmol, 72%).

LC/MS(C₂₉H₃₈N₆O₄SiS₂)627[M+H]⁺；RT 2.68(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.80(d,J＝7.5Hz,1H),7.49-7.38(m,2H),7.28-7.19(m,1H),5.85(s,2H),4.57(t,J＝5.2Hz,1H),4.37-4.22(m,4H),3.76-3.67(m,2H),3.53-3.44(m,2H),3.30-3.13(m,4H),2.32(s,3H),1.86-1.77(m,2H),1.31(t,J＝7.1Hz,3H),0.96-0.86(m,2H),-0.11(s,9H)。

Step D5- (3-chloropropyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

The product of step C (577mg,0.92mmol,1eq) was dissolved in thionyl chloride (30mL) and stirred at room temperature for 5 hours. The reaction was concentrated in vacuo, then partitioned between dichloromethane and brine, the organic phase dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a beige solid (341mg,0.53mmol, 57%).

LC/MS(C₂₉H₃₇ClN₆O₃SiS₂)645[M+H]⁺；RT 2.91(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.80(d,1H),7.50-7.39(m,2H),7.29-7.18(m,1H),5.85(s,2H),4.38-4.22(m,4H),3.78-3.67(m,4H),3.30-3.21(m,2H),2.32(s,3H),2.20-2.06(m,2H),1.31(t,J＝7.1Hz,2H),0.97-0.86(m,2H),-0.11(s,9H)。

Step E5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H, 7H-pyrrolo [2, 3-c)]Pyridazin-7-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried sealed flask was added a solution of the product of preparation 4b (133mg,0.69mmol,1.3eq) in dimethylformamide (70 mL). Sodium hydride (60% dispersion; 52.8mg,1.32mmol,2.5eq) was added to the solution and the mixture was stirred for 2 minutes. A solution of the product of step D (341mg,0.53mmol,1eq) in dimethylformamide (30mL) was added and the mixture was purged with nitrogen (10min) and heated at 100 ℃ for 1 h. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-7% methanol in dichloromethane afforded the desired product as a beige solid (203mg,0.25mmol, 48%).

LC/MS(C₄₀H₄₈FN₇O₄SiS₂)802[M+H]⁺；RT 2.59(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.79(d,1H),7.47-7.38(m,2H),7.31(dd,1H),7.27-7.11(m,3H),5.84(s,2H),4.36-4.20(m,4H),4.13(t,2H),3.75-3.66(m,2H),3.39(s,2H),3.31-3.19(m,4H),2.30(s,3H),2.19(s,6H),2.18-2.09(m,2H),1.28(t,3H),0.95-0.84(m,2H),-0.11(s,9H)。

Step F 2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl-)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step E (203mg,0.25mmol,1eq) in dichloromethane (10mL) was added trifluoroacetic acid (5.0mL,65.8mmol,260eq) and the mixture was stirred at room temperature for 6 h. Using dichloro chloride to the reaction solutionThe methane was diluted, cooled to 0 ℃ and neutralized by addition of 2M aqueous sodium hydroxide. The organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-5% methanol in dichloromethane afforded the desired product as a yellow solid (114mg,0.17mmol, 67%).

LC/MS(C₃₄H₃₄FN₇O₃S₂)672[M+H]⁺；RT 2.04(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.36(br s,1H),7.89(d,J＝7.9Hz,1H),7.58-7.47(m,1H),7.43-7.28(m,2H),7.26-7.12(m,3H),4.33(t,2H),4.27(q,2H),4.15(t,J＝6.1Hz,2H),3.40(s,2H),3.34-3.22(m,4H),2.33(s,3H),2.21(s,6H),2.16(t,2H),1.29(t,J＝7.1Hz,3H)。

Step G2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step F (114mg,0.17mmol,1eq) in 1, 4-dioxane (15mL) was added lithium hydroxide monohydrate (71.2mg,1.7mmol,10eq) and the mixture was heated at reflux for 7 h. The reaction was concentrated in vacuo and the residue triturated with water and acetonitrile, filtered and dried in vacuo to afford the desired product as a yellow solid (64.7mg,0.1mmol, 59%) as a lithium salt.

HRMS-ESI(m/z)[M+H]+C₃₂H₃₁FN₇O₃S₂644.1914, found 644.1908.

Example 233- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-7H-pyrrolo [2,3-c]Pyridazin-7-yl } pyridine-2-carboxylic acid

Step AN- (pent-3-yn-1-yl) carbamic acid tert-butyl ester

To a solution of pent-3-yn-1-amine hydrochloride (5g,41.8mmol,1eq) in tetrahydrofuran (130mL) and water (130mL) was added sodium bicarbonate (10.5g,125mmol,3eq) followed by di-tert-butyl dicarbonate (9.12g,41.8mmol,1eq) and the mixture was stirred at room temperature overnight. The reaction was diluted with ethyl acetate, washed successively with saturated aqueous sodium bicarbonate and brine, dried (magnesium sulfate) and concentrated in vacuo to afford the desired product as a yellow oil (8.2g, > 44.8mmol, > 100%).

¹H NMR(400MHz,DMSO-d6)δ6.89(t,J＝5.9Hz,1H),2.99(td,J＝7.3,5.9Hz,2H),2.25-2.15(m,2H),1.73(t,J＝2.6Hz,3H),1.38(s,9H)。

Step B3-bromo-6- { [ (tert-butoxy) carbonyl](pent-3-yn-1-yl) amino } pyridine-2-carboxylic acid ethyl ester

To an oven dried, sealed flask was added the product of step A (8.2g,44.8mmol,1eq), ethyl 3, 6-dibromopicolinate (13.8g,44.8mmol,1eq), Xantphos (2.59g,4.47mmol,0.1eq), cesium carbonate (29.2g,89.5mmol,2eq) and 1, 4-dioxane (180 mL). The vessel was evacuated and flushed with nitrogen, then tris (dibenzylideneacetone) dipalladium (0) (2.05g,2.24mmol,0.05eq) was added and the mixture purged with nitrogen (10min) and then heated at 130 ℃ for 1 hour. The reaction was diluted with ethyl acetate and filtered through celite, then washed sequentially with water and brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,330g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-8% ethyl acetate in isoheptane to give the desired product as a yellow oil (9.95g,24.2mmol, 54%).

LC/MS(C₁₈H₂₃BrN₂O₄)357[M-^tBu]⁺；RT 2.59(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.14(d,J＝8.9Hz,1H),7.71(d,J＝8.9Hz,1H),4.38(q,J＝7.1Hz,2H),3.96-3.87(m,2H),2.49-2.38(m,2H),1.66(t,J＝2.5Hz,3H),1.48(s,9H),1.33(t,J＝7.1Hz,3H)。

Step C3-bromo-6- [ (pent-3-yn-1-yl) amino]Pyridine-2-carboxylic acid ethyl ester

To the di-product of step B (9.95g,24.2mmol,1eq)To a solution of methyl chloride (120mL) was added trifluoroacetic acid (19.9mL,260mmol,10.8eq) and the mixture was stirred at room temperature overnight. The reaction was diluted with dichloromethane, cooled to 0 ℃ and neutralized by addition of 4M aqueous sodium hydroxide. The organic phase was dried (phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a yellow oil (6.68g,21.5mmol, 89%).

LC/MS(C₁₃H₁₅BrN₂O₂)313[M+H]⁺；RT 2.12(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.63(d,J＝9.0Hz,1H),7.20(t,J＝5.8Hz,1H),6.58(d,J＝9.0Hz,1H),4.32(q,J＝7.1Hz,2H),3.29(td,J＝7.1,5.7Hz,2H),2.41-2.29(m,2H),1.74(t,J＝2.6Hz,3H),1.30(t,J＝7.1Hz,3H)。

Step D3-bromo-6- { 3-chloro-4-methyl-7H-pyrrolo [2,3-c]Pyridazin-7-yl } pyridine-2-carboxylic acid ethyl ester

To a solution of the product of step C (6.68g,21.5mmol,1eq) in 1, 4-dioxane (220mL) was added 3, 6-dichloro-1, 2,4, 5-tetrazine (6.48g,42.9mmol,2eq) and the mixture was heated at 120 ℃ for 72 hours in a sealed flask. The reaction was diluted with methanol, filtered through a phase separator and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep) ^TMCartridge silica) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a red solid (1.49g,3.77mmol, 18%).

LC/MS(C₁₅H₁₂BrClN₄O₂)397[M+H]⁺；RT 2.36(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.93(d,J＝8.9Hz,1H),8.63(d,J＝3.9Hz,1H),8.53(d,J＝8.9Hz,1H),7.11(d,J＝3.9Hz,1H),4.45(q,J＝7.1Hz,2H),2.64(s,3H),1.37(t,3H)。

Step E3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- { 3-chloro-4-methyl-7H-pyrrolo [2, 3-c)]Pyridazin-7-yl } pyridine-2-carboxylic acid ethyl ester

To the product of step D (1.49g,3.77mmol,1eq) in tetrahydrofuran (5mL) and water (15mL) were added the product of preparation 5a (1.48g,4.14mmol,1.1eq) and potassium carbonate (1.56g,11.3mmol,3 eq). The vessel was evacuated and flushed with nitrogen, then Pd (dppf) Cl was added₂.CH₂Cl₂(308mg,0.38mmol,0.1eq) and the mixture was purged with nitrogen (10min) and then heated at 90 ℃ overnight in a sealed flask. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to afford the desired product as a white solid (1.08g,1.98mmol, 53%).

LC/MS(C₃₀H₃₃ClN₆O₂)545[M+H]⁺；RT 2.72(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.97(d,J＝8.6Hz,1H),8.69(d,J＝3.9Hz,1H),8.17(d,J＝8.6Hz,1H),7.43(s,1H),7.10(d,J＝3.9Hz,1H),4.22(q,J＝7.1Hz,2H),3.80(s,2H),2.65(s,3H),2.23(s,3H),2.03-1.89(m,3H),1.73-1.50(m,12H),1.16(t,3H)。

Step F3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-7H-pyrrolo [2,3-c]Pyridazin-7-yl } pyridine-2-carboxylic acid ethyl ester

To an oven dried sealed flask was added the product of step E (1.08mg,1.98mmol,1eq), 2-aminothiazole (594mg,3.96mmol,2eq), N-diisopropylethylamine (1.03mL,5.93mmol,3eq) and 1, 4-dioxane (80 mL). The vessel was evacuated and flushed with nitrogen, then JosiPhos (183mg,0.2mmol,0.1eq) was added and the mixture was purged with nitrogen (10min) and then heated at 150 ℃ for 2 days. The reaction was diluted with ethyl acetate, washed successively with water and brine, the organic phase dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to afford the desired product as an orange solid (510mg,0.77mmol, 39%).

LC/MS(C₃₇H₃₈N₈O₂S)659[M+H]⁺；RT 2.9(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ9.13(d,J＝8.6Hz,1H),8.61(d,J＝3.9Hz,1H),8.16(d,J＝8.6Hz,1H),7.91(br s,1H),7.62(br s,1H),7.44(s,1H),7.41-7.32(m,1H),7.27-7.11(m,1H),6.99(d,J＝3.9Hz,1H),4.22(q,J＝7.1Hz,2H),3.80(s,2H),2.65(s,3H),2.23(s,3H),2.03-1.89(m,3H),1.76-1.52(m,12H),1.16(t,3H)。

Step G3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-7H-pyrrolo [2,3-c]Pyridazin-7-yl } pyridine-2-carboxylic acid

To a solution of the product of step F (400mg,0.61mmol,1eq) in 1, 4-dioxane (15mL) was added lithium hydroxide monohydrate (255mg,6.07mmol,10eq) and the mixture was heated at reflux overnight. The reaction was concentrated in vacuo and the residue triturated with water, filtered and dried in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-14% methanol in dichloromethane afforded a light yellow solid which was triturated with methanol, filtered and dried in vacuo to afford the desired product as a yellow solid (154mg,0.24mmol, 40%).

HRMS-ESI(m/z)[M+H]+C₃₅H₃₅N₈O₂Calculated value of S631.2604, found value 631.2600.

Example 243- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } pyridine-2-carboxylic acid

Step A3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6-aminopyridine-2-carboxylic acid ethyl ester

A biphasic solution of ethyl 6-amino-3-bromopicolinate (5.14g,21mmol,1eq), the product of preparation 5a (7.47g,21mmol,1eq) and potassium carbonate (8.7g,62.9mmol,3eq) in tetrahydrofuran (100mL) and water (20mL) under nitrogenMix vigorously under a gas jet (10 min). Adding Pd (dppf) Cl₂.CH₂Cl₂(2.57g,3.15mmol,0.15eq) and the mixture was heated at reflux for 16 h. The reaction was cooled to room temperature and filtered through celite. The filtrate was diluted with ethyl acetate (200mL), washed with water (100mL) and the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-90% ethyl acetate in isoheptane to give the desired product as a cream solid (5.28g,13.4mmol, 64%).

LC/MS(C₂₃H₃₀N₄O₂)395[M+H]⁺；RT 1.32(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7032(d,1H),7.21(s,1H),6.58(d,1H),6.24(s,2H),4.05(q,2H),3.72(s,2H),2.10(s,3H),1.98-1.88(m,3H),1.71-1.47(m,12H),1.10(t,J＝7.1Hz,3H)。

Step B3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- { [3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl]Amino } pyridine-2-carboxylic acid ethyl ester

To a solution of the product of preparation 2e (1.92g,8.76mmol,1eq) and the product of step A (3.8g,9.64mmol,1.1eq) in methanol (40mL) was added acetic acid (15mL) and sodium cyanoborohydride (2.75g,43.8mmol,5eq) in portions and the mixture was heated at reflux for 1 h. The reaction was cooled to room temperature, then poured onto 1N aqueous sodium hydroxide (50mL) and extracted with ethyl acetate (3 × 50 mL). The organic phase was washed successively with saturated aqueous sodium bicarbonate (100mL) and brine (100mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to afford the desired product as a white solid (3.99g,6.68mmol, 76%).

LC/MS(C₃₁H₃₈Cl₂N₆O₂)597[M+H]⁺；RT 1.53(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.34(d,J＝8.6Hz,1H),7.22(s,1H),6.95(t,J＝5.7Hz,1H),6.63(d,J＝8.6Hz,1H),4.06(q,J＝7.1Hz,2H),3.72(s,2H),3.41-3.31(m,2H),2.92-2.83(m,2H),2.42(s,3H),2.10(s,3H),1.98-1.89(m,3H),1.86-1.74(m,2H),1.71-1.48(m,12H),1.08(t,J＝7.1Hz,3H)。

Step C3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- { 3-chloro-4-methyl-5H, 6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl } pyridine-2-carboxylic acid ethyl ester

To a solution of the product of step B (3.99g,6.68mmol,1eq) in α, α, α -trifluorotoluene (150mL) was added cesium carbonate (4.35g,13.4mmol,2eq) and the mixture was heated in a sealed tube at 160 ℃ for 3 days. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to afford the desired product as a white solid (0.95g,1.69mmol, 25%).

LC/MS(C₃₁H₃₇ClN₆O₂)561[M+H]⁺；RT 1.55(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.85(d,J＝8.6Hz,1H),7.71(d,J＝8.6Hz,1H),7.35(s,1H),4.15(q,J＝7.1Hz,2H),4.01-3.92(m,2H),3.77(s,2H),2.88(t,J＝6.6Hz,2H),2.31(s,3H),2.19(s,3H),2.04-1.88(m,5H),1.73-1.50(m,12H),1.13(t,J＝7.1Hz,3H)。

Step D3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } pyridine-2-carboxylic acid ethyl ester

To a solution of the product of step C (946mg,1.69mmol,1eq), 2-aminobenzothiazole (380mg,2.53mmol,1.5eq) and N, N-diisopropylethylamine (0.88mL,5.06mmol,3eq) in 1, 4-dioxane (30mL) was added Xantphos (97.6mg,0.17mmol,0.1eq) and tris (dibenzylideneacetone) dipalladium (0) (77.2mg,0.08mmol,0.05eq) and the mixture was heated in a sealed flask at 160 ℃ for 60 h. The reaction was cooled to room temperature and diluted with ethyl acetate (20mL), washed with brine (25mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80 gReddiSep) ^TMSilica gel cartridge) and gradient eluted with 0-75% ethyl acetate in isoheptane to giveDesired product as an orange solid (606mg,0.9mmol, 53%).

LC/MS(C₃₈H₄₂N₈O₂S)675[M+H]⁺；RT 1.61(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.95(d,J＝9.6Hz,1H),7.83(br s,1H),7.69(d,1H),7.57(br s,1H),7.40-7.31(m,2H),7.18(t,J＝7.5Hz,1H),4.15(q,J＝7.1Hz,2H),4.05-3.97(m,2H),3.77(s,2H),2.86(t,J＝6.5Hz,2H),2.33(s,3H),2.20(s,3H),2.04-1.89(m,5H),1.72-1.50(m,12H),1.13(t,3H)。

Step E3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } pyridine-2-carboxylic acid

To a solution of the product of step D (600mg,0.89mmol,1eq) in 1, 4-dioxane (10mL) was added lithium hydroxide monohydrate (74.6mg,1.78mmol,2eq) and the mixture was heated at reflux for 1 h. The reaction was cooled to room temperature and concentrated in vacuo. The residue was dissolved in water, acidified to pH 4 and the solid collected by filtration, washed with water and dried. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow solid (93mg,0.14mmol, 16%).

HRMS-ESI(m/z)[M+H]+C₃₆H₃₉N₈O₂Calculated value of S is 647.2917, found value is 647.2913.

Example 252- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- { [ (tert-butoxy) carbonyl](hept-5-yn-1-yl) amino } -1, 3-thiazole-4-carboxylic acid ethyl ester

To the 2- { [ (tert-butoxy) carbonyl group ]To a solution of ethyl amino } -1, 3-thiazole-4-carboxylate (1.62g,5.94mmol,1eq) in tetrahydrofuran (50mL) were added hept-5-yn-1-ol (1g,8.92mmol,1.5eq) and triphenylphosphine (2.34g,8.92mmol,1.5eq), then diethyl azodicarboxylate (1.62mL,8.92mmol,1.5eq) was added dropwise and the mixture was stirred at room temperature for 16 h. The reaction was partitioned between dichloromethane and brine, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMCartridge silica) and gradient eluted with 0-15% ethyl acetate in isoheptane to give the desired product as a yellow oil (1.92g,5.24mmol, 88%).

LC/MS(C₁₈H₂₆N₂O₄S)367[M+H]⁺；RT 2.53(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.07(s,1H),4.27(q,J＝7.1Hz,2H),4.11-4.00(m,2H),2.22-2.13(m,2H),1.79-1.68(m,5H),1.55(s,9H),1.48-1.34(m,2H),1.30(t,J＝7.1Hz,2H)。

Step B2- [ (hept-5-yn-1-yl) amino]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (1.97g,5.38mmol,1eq) in dichloromethane (50mL) was added trifluoroacetic acid (4.94mL,64.5mmol,12eq) and the mixture was stirred at room temperature overnight. The reaction was cooled to 0 ℃ and diluted with dichloromethane, basified with 2N aqueous sodium hydroxide, the organic phase dried (PTFE phase separator) and concentrated in vacuo to afford the desired product as a yellow oil (1.46g,5.48mmol, > 100%).

LC/MS(C₁₃H₁₈N₂O₂S)267[M+H]⁺；RT 1.91(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.81(t,J＝5.4Hz,1H),7.49(s,1H),5.77(s,1H),4.21(q,J＝7.1Hz,2H),3.21(td,J＝6.9,5.3Hz,2H),2.19-2.07(m,2H),1.73(t,J＝2.6Hz,3H),1.67-1.55(m,2H),1.54-1.41(m,2H),1.26(t,J＝7.1Hz,3H)。

Step C2- { 3-chloro-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b ] ]Aza derivatives-9-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (827mg,5.48mmol,1eq) in tetrahydrofuran (20mL) was added the product of step B (1.46g,5.48mmol,1eq) and the mixture was heated in a sealed flask at 120 ℃ for 48 h. The reaction was concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to afford the desired product as a pink solid (0.92g,2.61mmol, 48%).

LC/MS(C₁₅H₁₇ClN₄O₂S)353[M+H]⁺；RT 2.00(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.85(s,1H),4.28(q,J＝7.1Hz,3H),4.22-4.13(m,2H),2.95-2.88(m,2H),2.40(s,3H),2.02-1.89(m,2H),1.84-1.75(m,2H),1.30(t,J＝7.1Hz,3H)。

Step D2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step C (920mg,2.61mmol,1eq), 2-aminothiazole (470mg,3.13mmol,1.2eq), Xantphos (151mg,0.26mmol,0.1eq), N-diisopropylethylamine (1.36mL,7.82mmol,3eq) and 1, 4-dioxane (15 mL). The vessel was evacuated and flushed with nitrogen, then tris (dibenzylideneacetone) dipalladium (0) (119mg,0.13mmol,0.05eq) was added and the mixture was purged with nitrogen (10min) and then heated under microwave irradiation at 150 ℃ for 8 h. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give a solid which was triturated in acetonitrile, filtered and dried in vacuo to give the desired product as a yellow solid (0.81g,1.74mmol, 67%).

LC/MS(C₂₂H₂₂N₆O₂S₂)467[M+H]⁺；RT 2.20(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.88(br s,1H),7.75(s,1H),7.49(br s,1H),7.44-7.34(m,1H),7.27-7.16(m,1H),4.28(q,J＝7.1Hz,2H),4.14-4.01(s,2H),2.90-2.78(m,2H),2.42(s,3H),1.96-1.83(m,2H),1.82-1.70(m,2H),1.31(t,J＝7.1Hz,3H)。

Step E2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step D (30mg,0.06mmol,1eq) in 1, 4-dioxane (6mL) was added lithium hydroxide monohydrate (27mg,0.64mmol,10eq) and the mixture was heated at reflux for 6 h. The reaction was concentrated in vacuo, then dissolved in MeOH, applied to a short column of PE-AX (5g) wetted with methanol, washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane and methanol, filtered and dried in vacuo to give the desired product as an off-white solid (22.4mg,0.05mmol, 79%).

HRMS-ESI(m/z)[M+H]+C₂₀H₁₉N₆O₂S₂439.1011, found 439.1003.

Example 263- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl benzoic acids

Step A3- { [3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl]Amino } benzoic acid ethyl ester

To a solution of the product of preparation 2e (100mg,0.46mmol,1eq) and ethyl 3-aminobenzoate (79.2mg,0.48mmol,1.05eq) in methanol (6mL) was added acetic acid (2mL) and sodium cyanoborohydride (57.4mg,0.91mmol,2eq) in portions and the mixture was stirred overnight . The reaction was quenched by addition of 1N aqueous sodium hydroxide and extracted with ethyl acetate (3 × 50mL), and the combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a colorless oil (88mg,0.24mmol, 52%).

LC/MS(C₁₇H₁₉Cl₂N₃O₂)332[M-HCl+H]⁺；RT 1.34(LCMS-V-B1)

¹H NMR(400MHz,Chloroform-d)δ7.40(dt,J＝7.6,1.3Hz,1H),7.28(dd,J＝2.6,1.5Hz,1H),7.23(t,J＝7.9Hz,1H),6.78(ddd,J＝8.1,2.6,1.0Hz,1H),4.36(q,J＝7.2Hz,2H),3.32(t,J＝6.7Hz,2H),2.98-2.86(m,2H),2.40(s,3H),1.96-1.83(m,2H),1.38(t,J＝7.1Hz,3H)。

Step B3- { 3-chloro-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]]Pyridazin-8-yl benzoic acid ethyl ester

To a solution of the product of step A (88mg,0.24mmol,1eq) in dichloromethane (3mL) was added trifluoroacetic acid (0.1mL) and the mixture was stirred at room temperature overnight. The reaction was neutralized by addition of 1N aqueous sodium hydroxide solution and the mixture was extracted with dichloromethane (2 × 20 mL). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo to afford the desired product as a yellow solid (48mg,0.14mmol, 61%).

LC/MS(C₁₇H₁₈ClN₃O₂)332[M+H]⁺；RT 1.27(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.90(t,J＝1.9Hz,1H),7.80(dt,J＝7.6,1.5Hz,1H),7.66-7.60(m,1H),7.56(t,J＝7.7Hz,1H),4.33(q,J＝7.1Hz,2H),3.76-3.70(m,2H),2.87(t,J＝6.5Hz,2H),2.25(s,3H),2.09-1.97(m,2H),1.32(t,J＝7.1Hz,3H)。

Step C3- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl benzoic acid ethyl ester

To a solution of the product of step B (80mg,0.24mmol,1eq), 2-aminobenzothiazole (43.5mg,0.29mmol,1.2eq), N-diisopropylethylamine (0.13mL,0.72mmol,3eq), and 1, 4-dioxane (5mL) was added Xantphos (14mg,0.02mmol,0.1eq) and tris (dibenzylideneacetone) dipalladium (0) (11mg,0.01mmol,0.05eq) and the mixture was heated in a sealed tube at 160 ℃ for 24 h. The reaction was cooled to room temperature, then partitioned between ethyl acetate (20mL) and brine (25mL), the organic phase dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automatic flash chromatography (CombiFlash Rf, c 184.3 g RediSep column) with a gradient elution with 5-95% acetonitrile in water afforded the crude yellow glassy desired product (18mg,0.04mmol, 17%) which was used directly in the subsequent step without further purification.

LC/MS(C₂₄H₂₃N₅O₂S)446[M+H]⁺；RT 1.31(LCMS-V-B1)

Step D3- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl benzoic acids

To a solution of the product of step C (18mg,0.04mmol,1eq) in 1, 4-dioxane (5mL) was added lithium hydroxide monohydrate (3.39mg,0.08mmol,2eq) and the mixture was heated at reflux for 1 h. The reaction was cooled to room temperature and concentrated in vacuo. The residue was dissolved in water, acidified with 1N aqueous hydrochloric acid and concentrated in vacuo. Purification by preparative HPLC (HPLC-V-A1) gave the desired product as a yellow solid (10.6mg,0.03mmol, 63%).

HRMS-ESI(m/z)[M+H]+C₂₂H₂₀N₅O₂Calculated value of S is 418.1338, found value is 418.1334.

Example 272- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

General procedure for the preparation of propynylamine was used, starting from preparation 3d and dimethylamine as the appropriate amine. The desired product is then obtained using the general method of hydrolysis using the appropriate methyl ester as starting material.

HRMS-ESI(m/z):[M+H]+C₃₄H₃₅FN₇O₃S₂672.2221, found 672.2205.

Example 282- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-cyclopropyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

Step A2- { [ (tert-butoxy) carbonyl](4-iodobut-3-yn-1-yl) amino } -1, 3-thiazole-4-carboxylic acid ethyl ester

To the 2- { [ (tert-butoxy) carbonyl group]To a solution of ethyl amino } -1, 3-thiazole-4-carboxylate (3.16g,11.6mmol,1eq) in tetrahydrofuran (150mL) were added 4-iodobut-3-yn-1-ol (3.41g,17.4mmol,1.5eq) and triphenylphosphine (4.56g,17.4mmol,1.5eq), then diethyl azodicarboxylate (2.74mL,17.4mmol,1.5eq) was added dropwise and the mixture was stirred at room temperature for 16 hours. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-15% ethyl acetate in isoheptane to afford the desired product as a white solid (3.75g,8.33mmol, 72%).

LC/MS(C₁₅H₁₉IN₂O₄S)451[M+H]⁺；RT 2.45(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.09(s,1H),4.28(q,J＝7.1Hz,2H),4.16(t,J＝6.7Hz,2H),2.77(t,J＝6.7Hz,2H),1.56(s,9H),1.30(t,J＝7.1Hz,3H)。

Step B2- [ (4-iodobut-3-yn-1-yl) amino]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (3.75g,8.33mmol,1eq) in dichloromethane (50mL) was added trifluoroacetic acid (15.3mL,200mmol,24eq) and the mixture was stirred at room temperature for 1 h. The reaction was cooled to 0 ℃, diluted with dichloromethane, basified with 2N aqueous sodium hydroxide, the organic phase dried (PTFE phase separator) and concentrated in vacuo to afford the desired product as a white solid (2.74g,7.82mmol, 94%).

LC/MS(C₁₀H₁₁IN₂O₂S)351[M+H]⁺；RT 1.84(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.98(t,J＝5.7Hz,1H),7.53(s,1H),4.22(q,J＝7.1Hz,2H),3.40-3.30(m,2H),2.61(t,J＝6.8Hz,2H),1.26(t,J＝7.1Hz,3H)。

Step C2- { 3-chloro-4-iodo-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (1.18g,7.82mmol,1eq) in tetrahydrofuran (80mL) was added the product of step B (2.74g,7.82mmol,1eq) and the mixture was heated to reflux overnight. The reaction was cooled to room temperature and the precipitate was collected by filtration, washed with tetrahydrofuran and dried in vacuo to give the desired product as an off-white solid (1.06g,2.43mmol, 31%).

LC/MS(C₁₂H₁₀ClIN₄O₂S)437[M+H]⁺；RT 1.99(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.17(s,1H),4.45(t,J＝8.1Hz,2H),4.31(q,J＝7.1Hz,2H),3.46-3.33(m,2H),1.31(t,J＝7.1Hz,3H)。

Step D2- { 3-chloro-4-cyclopropyl-5H, 6H, 7H-pyrrolo [2,3-c ]]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a sealed tube was added the product of step C (120mg,0.27mmol,1eq), potassium cyclopropyltrifluoroborate (102mg,0.69mmol,2.5eq), potassium carbonate (114mg,0.82mmol,3eq), tetrahydrofuran (16mL) and water (4 mL). The vessel was evacuated and flushed with nitrogen, then Pd (dppf) Cl was added₂.CH₂Cl₂(44.9mg,0.05mmol,0.2eq) and the mixture was purged with nitrogen (10min) and then heated under microwave irradiation at 150 ℃ for 40 hours. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) purification with 0-90% ethyl acetate Gradient elution with isoheptane solution gave the desired product as a white solid (40mg,0.11mmol, 42%).

LC/MS(C₁₅H₁₅ClN₄O₂S)351[M+H]⁺；RT 2.00(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.12(d,J＝9.9Hz,1H),4.43-4.23(m,4H),3.42-3.29(m,2H),2.03-1.94(m,1H),1.30(t,3H),1.14-1.04(m,2H),0.98-0.84(m,2H)。

Step E2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-cyclopropyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried microwave vial was added the product of step D (40mg,0.11mmol,1eq), 2-aminothiazole (25.7mg,0.17mmol,1.5eq), N-diisopropylethylamine (59.6. mu.L, 0.34mmol,3eq) and 1, 4-dioxane (3 mL). The vessel was evacuated and flushed with nitrogen, then JosiPhos (10.5mg,0.01mmol,0.1eq) was added and the mixture was purged with nitrogen (10min) and then heated at 150 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to afford the desired product as a yellow solid (18mg,0.04mmol, 34%).

LC/MS(C₂₂H₂₀N₆O₂S₂)464[M+H]⁺；RT 2.31(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ10.58(br s,1H),8.06(s,1H),8.02-7.89(m,1H),7.76-7.62(m,1H),7.47-7.32(m,1H),7.30-7.13(m,1H),4.42-4.25(m,4H),3.46-3.37(m,2H),2.20-2.01(m,1H),1.32(t,J＝7.1Hz,3H),1.21-1.06(m,2H),0.93-0.74(m,2H)。

Step F2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-cyclopropyl-5H, 6H, 7H-pyrrolo [2,3-c]Pyridazin-7-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step E (18mg,0.04mmol,1eq) in 1, 4-dioxane (6mL) was added lithium hydroxide monohydrate (16.3mg,0.39mmol,10eq) and the mixture was heated at reflux for 8 h. The reaction was concentrated in vacuo, dissolved in methanol and then applied to a short column of PE-AX (10g) wetted with methanol, washed with methanol, eluted with 9:1 dichloromethane/formic acid and concentrated in vacuo. The residue was triturated with dichloromethane and water sequentially, filtered and dried in vacuo to give the desired product as a beige solid (2.44mg,0.01mmol, 14.43%) as the formate salt.

HRMS-ESI(m/z)[M+H]+C₂₀H₁₇N₆O₂S₂437.0854, found 437.0853.

Example 293- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } pyridine-2-carboxylic acid

Step A[ (hex-5-yn-1-yloxy) methyl group]Benzene and its derivatives

To a stirred solution of 5-hexyn-1-ol (5.36g,54.6mmol,1eq) in tetrahydrofuran (35mL) cooled to 0 deg.C was added sodium hydride (60% dispersion; 3.28g,81.9mmol,1.5eq) in portions and the mixture was stirred for 30 min. Benzyl bromide (6.49mL,54.6mmol,1eq) was added dropwise and the mixture was warmed to room temperature and stirred for 90 h. The reaction was cooled to 0 ℃ and quenched by the addition of saturated aqueous ammonium chloride (30mL) and then diluted with water (30 mL). The mixture was extracted with ethyl acetate (2 × 150mL), the combined organic extracts were washed with brine (100mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% ethyl acetate in isoheptane to give the desired product as a yellow oil (10.2g,54.2mmol, 99%).

LC/MS(C₁₃H₁₆O)189[M+H]⁺；RT 2.21(LCMS-V-C)

¹H NMR(400MHz,Chloroform-d)δ7.40-7.24(m,5H),4.45(s,2H),3.44(t,J＝6.3Hz,2H),2.77(t,J＝2.7Hz,1H),2.17(td,J＝7.0,2.6Hz,2H),1.70-1.57(m,2H),1.56-1.37(m,2H)。

Step B[ (hept-5-yn-1-yloxy) methyl group]Benzene and its derivatives

The product of step A (10.2g,54.2mmol,1eq) in tetrahydrofuran (90mL) was cooled to-78 deg.C and n-butyllithium (2.5M in hexane; 26mL,65mmol,1.2eq) was added dropwise over 30 minutes. After stirring for 1 hour, iodomethane (4.05mL,65mmol,1.2eq) was added dropwise and the mixture was warmed to 0 ℃ over 1 hour. The reaction was quenched with saturated aqueous ammonium chloride (40mL), diluted with water (40mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic extracts were washed successively with 2N aqueous sodium thiosulfate (200mL) and brine (200mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep) ^TMCartridge silica gel) and gradient eluted with 0-6% ethyl acetate in isoheptane to give the desired product as a clear oil (10.4g,51.3mmol, 95%).

LC/MS(C₁₄H₁₈O)203[M+H]⁺；RT 2.37(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.40-7.24(m,5H),4.45(s,2H),3.43(t,J＝6.4Hz,2H),2.17-2.08(m,2H),1.73(t,J＝2.6Hz,3H),1.67-1.55(m,2H),1.54-1.41(m,2H)。

Step C4- [4- (benzyloxy) butyl]-3, 6-dichloro-5-methylpyridazine

A solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (3.23g,21.4mmol,1eq) and the product of step B (5.2g,25.7mmol,1.2eq) in toluene (40mL) was heated in a sealed flask at 130 ℃ overnight. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a red oil (3.27g,10.1mmol, 47%).

LC/MS(C₁₆H₁₈Cl₂N₂O)325[M+H]⁺；RT 2.32(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.40-7.24(m,5H),4.46(s,2H),3.49(t,J＝6.1Hz,2H),2.87-2.78(m,2H),2.41(s,3H),1.74-1.62(m,2H),1.61-1.51(m,2H)。

Step D4- (3, 6-dichloro-5-methylpyridazin-4-yl) butan-1-ol

To a solution of the product of step C (3.27g,10.1mmol,1eq) in dichloromethane (50mL) cooled in an ice-water bath was added boron trichloride (1M in dichloromethane; 50.3mL,50.3mmol,5eq) dropwise and the mixture was warmed to room temperature and stirred for 1 hour. The reaction was cooled to 0 ℃, quenched by addition of methanol and concentrated in vacuo. The residue was partitioned between dichloromethane (100mL) and saturated aqueous sodium bicarbonate (150mL) and the organic phase was washed with brine (150mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-80% ethyl acetate in isoheptane to give the desired product as a yellow oil (2.21g,9.4mmol, 94%).

LC/MS(C₉H₁₂Cl₂N₂O)235[M+H]⁺；RT 1.36(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ4.44(t,J＝5.1Hz,1H),3.45(dd,J＝6.0,5.0Hz,2H),2.87-2.76(m,2H),2.43(s,3H),1.62-1.48(m,4H)。

Step E4- (3, 6-dichloro-5-methylpyridazin-4-yl) butanal

To an oven dried flask was added dimethyl sulfoxide (1.6mL,22.6mmol,2.4eq) and dichloromethane (60mL) and the mixture was cooled to-78 ℃. Oxalyl chloride (2M in dichloromethane; 7.05mL,14.1mmol,1.5eq) was added dropwise and the mixture was stirred for 1 hour. A solution of the product of step D (2.21g,9.4mmol,1eq) in dichloromethane (20mL) was added dropwise and the mixture was stirred for 1 hour. Triethylamine (7.84mL,56.4mmol,6eq) was added and the mixture was warmed to 0 ℃ over 1 hour. The reaction was quenched with water (50mL), diluted with saturated aqueous sodium bicarbonate (50mL) and extracted with dichloromethane (2x200 mL). The combined organic extracts were washed with brine (100mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to giveDesired product as yellow oil (6.58g,6.78mmol, 72%).

LC/MS(C₉H₁₀Cl₂N₂O)233[M+H]⁺；RT 1.51(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ9.70(t,J＝1.1Hz,1H),2.86-2.76(m,2H),2.63(td,J＝7.0,1.1Hz,2H),2.45(s,3H),1.81-1.68(m,2H)。

Step F3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- { [4- (3, 6-dichloro-5-methylpyridazin-4-yl) butyl ]Amino } pyridine-2-carboxylic acid ethyl ester

To a solution of the product of step E (1.04g,4.45mmol,1eq) and the product of example 24, step A (1.93g,4.89mmol,1.1eq) in methanol (30mL) and acetic acid (10mL) was added sodium cyanoborohydride (559mg,8.89mmol,2eq) and the mixture was stirred at room temperature overnight. The reaction was quenched with 1N aqueous sodium hydroxide (50mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic extracts were washed successively with saturated aqueous sodium bicarbonate and brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to give the desired product as a white gum (1.71g,2.8mmol, 63%).

LC/MS(C₃₂H₄₀Cl₂N₆O₂)611[M+H]⁺；RT 2.65(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.32(d,J＝8.6Hz,1H),7.21(s,1H),6.81(t,J＝5.4Hz,1H),6.60(d,J＝8.6Hz,1H),4.05(q,J＝7.1Hz,2H),3.72(s,2H),3.32-3.23(m,2H),2.90-2.81(m,2H),2.43(s,3H),2.10(s,3H),1.94(s,3H),1.74-1.48(m,16H),1.07(t,J＝7.1Hz,3H)。

Step G3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- { 3-chloro-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } pyridine-2-carboxylic acid ethyl ester

To the product of step F (646mg,1.06mmol,1eq) was added α, αCesium carbonate (1.03G,3.17mmol,3eq) and XantPhos Pd G3(50.1mg,0.05mmol,0.05eq) were added to a solution of α -trifluorotoluene (6mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ overnight. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to afford the desired product as an off white solid (119mg,0.21mmol, 20%).

LC/MS(C₃₂H₃₉ClN₆O₂)575[M+H]⁺；RT 2.66(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.47(d,J＝8.8Hz,1H),7.29(s,1H),6.80(d,J＝8.7Hz,1H),4.22-4.16(m,2H),4.12(q,2H),3.73(s,2H),2.89-2.81(m,2H),2.45(s,3H),2.14(s,3H),2.00-1.89(m,3H),1.80-1.49(m,16H),1.10(t,3H)。

Step H3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } pyridine-2-carboxylic acid ethyl ester

To a solution of the product of step G (119mg,0.21mmol,1eq), 2-aminobenzothiazole (62.2mg,0.41mmol,2eq) and N, N-diisopropylethylamine (108 μ L,0.62mmol,3eq) in 1, 4-dioxane (8mL) was added JosiPhos (19.2mg,0.02mmol,0.1eq) and the mixture was heated in a sealed tube at 150 ℃ for 72 h. The reaction was cooled to room temperature, then diluted with ethyl acetate (20mL), washed with brine (25mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the crude desired product as a yellow gum (49mg,0.07mmol, 34%) which was used directly in the next step without further purification.

LC/MS(C₃₉H₄₄N₈O₂S)690[M+H]⁺；RT 2.81(LCMS-V-C)

Step I3- {1- [ (adamantan-1-yl) methyl group]-5-methyl-1H-pyrazol-4-yl } -6- {3- [ (1, 3-benzothiazol-2-yl) amino group ]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b ]]Aza derivatives-9-yl } pyridine-2-carboxylic acid

To a solution of the product of step H (49mg,0.07mmol,1eq) in 1, 4-dioxane (15mL) was added lithium hydroxide monohydrate (44.8mg,1.07mmol,15eq) and the mixture was heated at reflux for 1H. The reaction was cooled to room temperature and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, c184.3g RediSep column) with a gradient of 5-95% acetonitrile in water afforded the desired product as a yellow solid (6.24mg,0.01mmol, 13%).

HRMS-ESI(m/z)[M+H]+C₃₇H₄₁N₈O₂Calculated value of S661.3073, found value of 661.3097

Example 302- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-cyclopropyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step AT-butyl [ (5-cyclopropylpent-4-yn-1-yl) oxy ] group]Dimethylsilane

To a solution of cyclopropylacetylene (8mL,94.5mmol,1.1eq) in tetrahydrofuran (200mL) cooled to-78 deg.C was added n-butyllithium (2.0M in hexane; 47.3mL,94.5mmol,1.1eq) and the mixture was stirred at that temperature for 2.5 h. 1, 3-dimethyl-3, 4,5, 6-tetrahydro-2 (1H) -pyrimidinone (12mL,98.8mmol,1.15eq) was added and after 15 min (3-bromopropoxy) -tert-butyldimethylsilane (15mL,85.9mmol,1eq) was added dropwise and the mixture was warmed to room temperature and stirred overnight. The reaction was partitioned between ethyl acetate and saturated aqueous ammonium chloride, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic quick-acting Rapid column chromatography (Combiflash Rf,330g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-6% ethyl acetate in isoheptane to give crude desired product as a clear oil (8.62g,36.2mmol, 42%) which was used directly in the next step without further purification.

Step B5-cyclopropylpent-4-yn-1-ol

To a solution of the product of step A (8.62g,36.2mmol,1eq) in tetrahydrofuran (150mL) was added tetrabutylammonium fluoride (1M in tetrahydrofuran; 39.8mL,39.8mmol,1.1eq) and the mixture was stirred at room temperature for 1 hour. The reaction was concentrated in vacuo, partitioned between ethyl acetate and water, the organic phase dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a clear oil (2.14g,17.2mmol, 48%).

¹H NMR(400MHz,DMSO-d6)δ4.43(t,J＝5.2Hz,1H),3.46-3.38(m,2H),2.12(td,J＝7.1,2.0Hz,2H),1.58-1.47(m,2H),1.29-1.16(m,1H),0.74-0.65(m,2H),0.54-0.46(m,2H)。

Step C2- { [ (tert-butoxy) carbonyl](5-Cyclopropylpent-4-yn-1-yl) amino } -1, 3-thiazole-4-carboxylic acid ethyl ester

To 2- [ (tert-butoxycarbonyl) amino group]To a solution of ethyl-1, 3-thiazole-4-carboxylate (3.13g,11.5mmol,1eq) and the product of step B (2.14g,17.2mmol,1.5eq) in tetrahydrofuran (80mL) was added polymer-supported triphenylphosphine (4.52g,17.23mmol,1.5eq), the mixture was cooled to 0 ℃ and diethyl azodicarboxylate (2.73mL,17.2mmol,1.5eq) was added dropwise, then the mixture was warmed to room temperature and stirred overnight. The reaction was partitioned between dichloromethane and water, separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-15% ethyl acetate in isoheptane to give the desired product as a colorless solid (3.86g,10.2mmol, 89%).

LC/MS(C₁₉H₂₆N₂O₄S)379[M+H]⁺；RT 2.60(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ8.07(s,1H),4.28(q,J＝7.1Hz,2H),4.11(t,J＝7.0Hz,2H),2.21-2.13(m,2H),1.84-1.75(m,2H),1.58(s,9H),1.55(s,4H),1.29(t,3H),1.23-1.14(m,1H),0.73-0.61(m,2H),0.56-0.45(m,2H)。

Step D2- { 3-chloro-4-cyclopropyl-5H, 6H,7H, 8H-pyrido [2,3-c ]]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step C (3.86g,10.2mmol,1eq) in toluene (120mL) was added 3, 6-dichloro-1, 2,4, 5-tetrazine (1.54g,10.2mmol,1eq) and the mixture was heated in a sealed flask at 130 ℃ for 24 h. The reaction was concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to give the desired product (677mg,1.86mmol, 18%).

LC/MS(C₁₆H₁₇ClN₄O₂S)365[M+H]⁺；RT 2.18(LCMS-V-C)

¹H NMR(400MHz,TFA added/DMSO-d6)δ8.04(s,1H),4.36-4.22(m,4H),3.07(t,J＝6.2Hz,2H),2.11-2.00(m,2H),1.92-1.81(m,1H),1.31(t,J＝7.1Hz,3H),1.20-1.06(m,2H),0.76-0.67(m,2H)。

Step E2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-cyclopropyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried sealed tube was added the product of step D (627mg,1.72mmol,1eq), 2-aminobenzothiazole (387mg,2.58mmol,1.5eq), N-diisopropylethylamine (0.9mL,5.16mmol,3eq) and 1, 4-dioxane (22mL) and the mixture was purged with nitrogen (10min) then Josiphos Pd G3(162mg,0.17mmol,0.1eq) was added and the mixture was heated at 150 ℃ for 20 h. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a beige solid (261mg,0.55mmol, 32%). LC/MS (C)₂₃H₂₂N₆O₂S₂)479[M+H]⁺；RT 2.44(LCMS-V-C)

¹H NMR(400MHz,TFA added/DMSO-d6)δ10.45(s,1H),8.07-7.92(m,2H),7.75-7.60(m,1H),7.40(t,1H),7.30-7.16(m,1H),4.38-4.22(m,4H),3.08(t,J＝6.1Hz,2H),2.15-2.01(m,2H),1.98-1.85(m,1H),1.32(t,J＝7.1Hz,3H),1.27-1.12(m,2H),0.74-0.53(m,2H)。

Step F2- {3- [ (1, 3-benzothiazol-2-yl) ({ [2- (trimethylsilyl) ethoxy)]Methyl }) amino]-4-cyclopropyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step E (1.47g,3.07mmol,1eq) in dimethylformamide (240mL) was added N, N-diisopropylethylamine (1.61mL,9.21mmol,3eq), after 5 min the mixture was cooled to 0 ℃ and 4- (dimethylamino) pyridine (75.1mg,0.61mmol,0.2eq) and 2- (trimethylsilyl) ethoxymethyl chloride (1.62mL,9.21mmol,3eq) were added and the mixture was warmed to room temperature and stirred overnight. The reaction was concentrated in vacuo, partitioned between dichloromethane and brine, separated (PTFE phase separator) and the organic phase concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a yellow gum (1.36g,2.23mmol, 73%).

LC/MS(C₂₉H₃₆N₆O₃SiS₂)609[M+H]⁺；RT 2.96(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.97(s,1H),7.80(d,1H),7.48-7.37(m,2H),7.22(ddd,J＝8.3,7.0,1.5Hz,1H),5.82(s,2H),4.36-4.21(m,4H),3.76-3.66(m,2H),3.06(t,J＝6.2Hz,2H),2.13-2.02(m,2H),1.95-1.84(m,1H),1.31(t,J＝7.1Hz,3H),1.23-1.15(m,2H),1.13-1.05(m,2H),0.93-0.84(m,2H),-0.10(s,9H)。

Step G2- {3- [ (1, 3-benzothiazol-2-yl) ({ [2- (trimethylsilyl) ethoxy) ]Methyl }) amino]-4-cyclopropyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5-bromo-1, 3-thiazole-4-carboxylic acid ethyl ester

To the product of step F (1.36g,2.23mmol,1eq) in dichloromethane (40mL) was dissolvedN-bromosuccinimide (596mg,3.35mmol,1.5eq) was added to the solution and the mixture was stirred at room temperature overnight. The reaction was partitioned between dichloromethane and brine, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to afford the desired product as a yellow solid (1.43g,2.08mmol, 93%).

LC/MS(C₂₉H₃₅BrN₆O₃SiS₂)689[M+H]⁺；RT 3.17(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.86-7.81(m,1H),7.50-7.39(m,2H),7.27-7.20(m,1H),5.82(s,2H),4.32(q,2H),4.29-4.21(m,2H),3.75-3.66(m,2H),2.13-2.01(m,2H),1.98-1.86(m,1H),1.32(t,3H),1.28-1.16(m,2H),1.15-1.05(m,2H),0.94-0.83(m,2H),-0.10(s,9H)。

Step H5- [ (1E) -3- [ (tert-butyldimethylsilyl) oxy group]Prop-1-en-1-yl]-2- (4-cyclopropyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried sealed flask was added the product of step G (1.43G,2.08mmol,1eq), (E) -3- (tert-butyldimethylsilyloxy) propen-1-yl-boronic acid pinacol ester (0.82mL,2.5mmol,1.2eq), potassium carbonate (862mg,6.24mmol,3eq), [1,1' -bis (diphenylphosphino) ferrocene ]Palladium (II) dichloride (170mg,0.21mmol,0.1eq), tetrahydrofuran (60mL) and water (20mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ for 1.5 h. The reaction was partitioned between ethyl acetate and water, the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to afford the desired product as a yellow solid (1.05g,1.35mmol, 65%).

LC/MS(C₃₈H₅₄N₆O₄Si₂S₂)779[M+H]⁺；RT 1.66(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.79(d,1H),7.51-7.37(m,3H),7.27-7.18(m,1H),6.27(dt,J＝16.0,4.3Hz,1H),5.82(s,2H),4.40-4.34(m,2H),4.34-4.23(m,4H),3.75-3.66(m,2H),3.06(t,J＝6.1Hz,2H),2.13-2.01(m,2H),1.96-1.84(m,1H),1.31(t,J＝7.1Hz,3H),1.22-1.14(m,2H),1.13-1.04(m,2H),0.93(s,9H),0.89-0.82(m,2H),0.11(s,6H),-0.10(s,9H)。

Step I5- {3- [ (tert-butyldimethylsilyl) oxy group]Propyl } -2- (4-cyclopropyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step H (1.05g,1.35mmol,1eq) in ethyl acetate (60mL) under nitrogen was added platinum (IV) oxide (91.8mg,0.4mmol,0.3 eq). The vessel was evacuated and backfilled with nitrogen (x3), then evacuated, placed under a hydrogen atmosphere and shaken at room temperature for 2 days. The reaction was filtered through celite, eluted with ethyl acetate and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMCartridge silica) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a cream gum (913mg,1.17mmol, 87%).

LC/MS(C₃₈H₅₆N₆O₄Si₂S₂)781[M+H]⁺；RT 1.71(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.74(d,1H),7.49-7.36(m,2H),7.27-7.20(m,1H),5.82(s,2H),4.34-4.21(m,4H),3.75-3.62(m,4H),3.15(t,J＝7.5Hz,2H),3.05(t,J＝6.3Hz,2H),2.12-2.00(m,2H),1.96-1.78(m,3H),1.31(t,3H),1.21-1.16(m,2H),1.13-1.05(m,2H),0.91(s,9H),0.87-0.81(m,2H),0.06(s,6H),-0.10(m,9H)。

Step J2- (4-cyclopropyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- (3-hydroxypropyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To the product of step I (819mg,1.05mmol,1eq) in 1, 4-dioxane (18 mL)) Tetrabutylammonium fluoride (1M solution in tetrahydrofuran; 1.15mL,1.15mmol,1.1eq) and the mixture was stirred at room temperature for 2 hours. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in dichloromethane afforded the desired product as a yellow gum (650mg,0.97mmol, 93%).

LC/MS(C₃₂H₄₂N₆O₄SiS₂)667[M+H]⁺；RT 2.85(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.81(dd,J＝7.5,1.0Hz,1H),7.48-7.36(m,2H),7.27-7.17(m,1H),5.81(s,2H),4.56(t,J＝5.1Hz,1H),4.34-4.21(m,4H),4.12(q,J＝7.1Hz,3H),3.75-3.66(m,2H),3.48(td,J＝6.3,5.1Hz,2H),3.17-3.08(m,2H),3.05(t,J＝6.3Hz,2H),1.96-1.84(m,1H),1.83-1.74(m,2H),1.31(t,J＝7.1Hz,3H),1.20-1.15(m,2H),1.13-1.04(m,2H),0.93-0.85(m,2H),-0.10(s,9H)。

Step K5- (3-chloropropyl) -2- (4-cyclopropyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester the product of step J (291mg,0.44mmol,1eq) was dissolved in thionyl chloride (10mL) and stirred at room temperature for 8 h. The reaction was concentrated in vacuo, then partitioned between dichloromethane and brine, the organic phase dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMCartridge silica) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as an orange gum (147mg,0.21mmol, 49%).

LC/MS(C₃₂H₄₁ClN₆O₃SiS₂)685[M+H]⁺；RT 3.15(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.80(dd,J＝7.4,1.0Hz,1H),7.48-7.36(m,2H),7.27-7.18(m,1H),5.81(s,2H),4.34-4.21(m,4H),3.78-3.64(m,4H),3.27-3.18(m,2H),3.05(t,J＝6.2Hz,2H),2.16-1.96(m,4H),1.98-1.81(m,1H),1.32(t,J＝7.1Hz,3H),1.22-1.14(m,2H),1.13-1.01(m,2H),0.95-0.82(m,2H),-0.11(s,9H)。

Step L2- (4-cyclopropyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of preparation 4b (53.9mg,0.28mmol,1.3eq) in dimethylformamide (30mL) was added sodium hydride (60% dispersion; 21.5mg,0.54mmol,2.5eq) and the mixture was stirred for 2 min. A solution of the product of step K (147mg,0.21mmol,1eq) in dimethylformamide (10mL) was added and the mixture was heated at 100 ℃ for 1.5 h. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-5% methanol in dichloromethane afforded the desired product as a yellow gum (145mg,0.17mmol, 80%).

LC/MS(C₄₃H₅₂FN₇O₄SiS₂)842[M+H]⁺；RT 2.76(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.79(dd,J＝7.5,1.1Hz,1H),7.48-7.36(m,2H),7.34-7.27(m,1H),7.26-7.12(m,3H),5.81(s,2H),4.30-4.20(m,4H),4.14(t,J＝6.1Hz,2H),3.75-3.66(m,2H),3.38(s,2H),3.27(t,J＝6.3Hz,2H),3.05(t,2H),2.19(s,6H),2.17-2.09(m,2H),2.08-1.99(m,2H),1.94-1.83(m,1H),1.29(t,3H),1.21-1.14(m,2H),1.13-1.05(m,2H),0.94-0.82(m,2H),-010(s,9H)。

Step M2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-cyclopropyl-5H, 6H,7H, 8H-pyrido [2,3-c ]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl-)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester to a solution of the product of step L (175mg,0.21mmol,1eq) in dichloromethane (6mL) was added trifluoroacetic acid (6mL) and the mixture was stirred at room temperature for 7.5 h. The reaction solution was diluted with dichloromethane, cooled to 0 ℃ and neutralized with 2N aqueous sodium hydroxide solution. The organic phase is dried (PTFE phase separation)Vessel) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-7% methanol in dichloromethane afforded the desired product as a yellow gum (16mg,0.02mmol, 11%).

LC/MS(C₃₇H₃₈FN₇O₃S₂)712[M+H]⁺；RT 2.18(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.94(d,J＝7.7Hz,1H),7.60(d,J＝8.0Hz,1H),7.43-7.34(m,1H),7.30(dd,J＝11.9,2.0Hz,1H),7.26-7.10(m,3H),4.30-4.20(m,4H),4.14(t,J＝6.1Hz,2H),3.36(s,2H),3.30-3.23(m,2H),3.10-3.00(m,2H),2.19(s,6H),2.16-2.09(m,2H),2.08-1.99(m,2H),1.95-1.86(m,1H),1.28(t,J＝7.1Hz,3H),1.25-1.21(m,2H),1.21-1.13(m,2H)。

Step N2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-cyclopropyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step M (16mg,0.02mmol,1eq) in 1, 4-dioxane (5mL) was added lithium hydroxide monohydrate (9.43mg,0.22mmol,10eq) and the mixture was heated at reflux overnight. The reaction was concentrated in vacuo and the residue triturated with water then diethyl ether, filtered and dried in vacuo to give the desired product as a yellow solid (10.4mg,0.02mmol, 68%) as the lithium salt.

HRMS-ESI(m/z)[M+H]+C₃₅H₃₅FN₇O₃S₂684.2227, found 684.2223.

Example 312- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H,8H, 9H-pyridazino [3, 4-b)]Aza derivatives-9-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of example 25(4.97g,10.7mmol,1eq) in dimethylformamide (180mL) was added N, N-diisopropylethylamine (5.57mL,32mmol,3 eq). After 5 min the mixture was cooled to 0 ℃,4- (dimethylamino) pyridine (260mg,2.13mmol,0.2eq) and 2- (trimethylsilyl) ethoxymethyl chloride (5.61mL,32mmol,3eq) were added and the mixture was stirred at room temperature overnight. The reaction was concentrated in vacuo, partitioned between ethyl acetate and water, the organic phase dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a yellow gum (5.21g,8.73mmol, 82%).

LC/MS(C₂₈H₃₆N₆O₃SiS₂)597[M+H]⁺；RT 2.87(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dd,J＝7.7,1.1Hz,1H),7.74(s,1H),7.52-7.39(m,2H),7.25(td,J＝7.5,1.4Hz,1H),5.88(s,2H),4.27(q,J＝7.1Hz,2H),4.15-4.01(m,2H),3.72(dd,J＝8.4,7.4Hz,2H),2.89-2.81(m,2H),2.43(s,3H),1.95-1.83(m,2H),1.82-1.69(m,2H),1.29(t,J＝7.1Hz,3H),0.91(dd,J＝8.5,7.4Hz,2H),-0.11(s,9H)。

Step B5-bromo-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H,8H, 9H-pyridazino [3, 4-b)]Aza derivatives-9-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To the product of step A (5.21g,8.73mmol,1eq) in dichloromethane (100mL) was added N-bromosuccinimide (1.71g,9.6mmol,1.1eq) and the mixture was stirred at room temperature overnight. The reaction was partitioned between dichloromethane and 10% aqueous sodium thiosulfate, the organic phase was washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a yellow gum (5.23g,7.74mmol, 89%).

LC/MS(C₂₈H₃₅BrN₆O₃SiS₂)677[M+H]⁺；RT 3.08(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.84(dd,J＝7.9,1.1Hz,1H),7.52-7.38(m,2H),7.31-7.20(m,1H),5.88(s,2H),4.30(q,J＝7.1Hz,2H),4.18(dd,J＝6.8,4.6Hz,2H),3.78-3.66(m,2H),2.95-2.85(m,2H),2.44(s,3H),2.01-1.88(m,2H),1.86-1.72(m,2H),1.31(t,J＝7.1Hz,3H),0.96-0.87(m,2H),-0.10(s,9H)。

Step C5- [ (1E) -3- [ (tert-butyldimethylsilyl) oxy group]Prop-1-en-1-yl]-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H,8H, 9H-pyridazino [3, 4-b)]Aza derivatives-9-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To an oven dried sealed flask was added the product of step B (5.23g,7.74mmol,1eq), (E) -3- (tert-butyldimethylsilyloxy) propen-1-yl-boronic acid pinacol ester (3.04mL,9.29mmol,1.2eq), potassium carbonate (3.21g,23.2mmol,3eq), [1,1' -bis (diphenylphosphino) ferrocene ]Palladium (II) dichloride (632mg,0.77mmol,0.1eq), tetrahydrofuran (150mL) and water (50mL) and the mixture was purged with nitrogen (10min) and then heated at 120 ℃ for 1 hour. The reaction was partitioned between ethyl acetate and water, the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give orangeDesired product as a colored oil (5.17g,6.74mmol, 87%).

LC/MS(C₃₇H₅₄N₆O₄Si₂S₂)767[M+H]⁺；RT 1.60(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.82(dd,J＝7.8,1.2Hz,1H),7.53-7.35(m,3H),7.26(ddd,J＝8.3,7.2,1.3Hz,1H),6.01(dt,J＝15.9,4.3Hz,1H),5.88(s,2H),4.36-4.235(m,4H),4.12-3.99(m,2H),3.78-3.67(m,2H),2.89-2.78(m,2H),2.43(s,3H),1.95-1.82(m,2H),1.81-1.68(m,2H),1.30(t,J＝7.1Hz,3H),0.97-0.84(m,11H),0.07(s,6H),-0.12(s,9H)。

Step D5- {3- [ (tert-butyldimethylsilyl) oxy group]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H,8H, 9H-pyridazino [3, 4-b)]Aza derivatives-9-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step C (5.17g,6.74mmol,1eq) in ethyl acetate (120mL) under nitrogen was added platinum (IV) oxide (459mg,2.02mmol,0.3 eq). The vessel was evacuated and backfilled with nitrogen (x3), then evacuated, placed under a hydrogen atmosphere and shaken at room temperature for 2 days. The reaction was filtered through celite, eluted with ethyl acetate and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep) ^TMCartridge silica gel) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as an orange oil (4.46g,5.8mmol, 86%).

LC/MS(C₃₇H₅₆N₆O₄Si₂S₂)769[M+H]⁺；RT 1.62(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.79(dd,J＝7.9,1.2Hz,1H),7.53-7.40(m,2H),7.31-7.21(m,1H),5.87(s,2H),4.26(q,J＝7.1Hz,2H),4.10-3.97(m,2H),3.77-3.66(m,2H),3.61(t,J＝6.1Hz,2H),3.60(t,J＝7.6Hz,2H),3.11-2.99(m,2H),2.88-2.76(m,2H),2.44(s,3H),1.96-1.81(m,2H),1.81-1.67(m,4H),1.29(t,3H),0.91-0.81(m,11H),0.01(s,6H),-0.11(s,9H)。

Step E5- (3-hydroxypropyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H,8H, 9H-pyridazino [3, 4-b)]Aza derivatives-9-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step D (4.46g,5.8mmol,1eq) in tetrahydrofuran (150mL) was added tetrabutylammonium fluoride (1M in tetrahydrofuran, 8.7mL,8.7mmol,1.5eq) and the mixture was stirred at room temperature for 1.5 h. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in dichloromethane afforded the desired product as a beige gum (2.63g,4.02mmol, 69%).

LC/MS(C₃₁H₄₂N₆O₄SiS₂)655[M+H]⁺；RT 2.77(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.83(dd,J＝7.9,1.2Hz,1H),7.52-7.39(m,2H),7.25(ddd,J＝8.3,7.1,1.3Hz,1H),5.88(s,2H),4.50(t,J＝5.1Hz,1H),4.26(q,J＝7.1Hz,2H),4.07-3.98(m,2H),3.72(t,J＝7.9Hz,2H),3.42(td,J＝6.3,5.1Hz,2H),3.04(dd,J＝8.9,6.4Hz,2H),2.90-2.77(m,2H),2.43(s,3H),1.95-1.82(m,2H),1.81-1.65(m,4H),1.29(t,J＝7.1Hz,3H),0.91(dd,J＝8.4,7.4Hz,2H),-0.12(s,9H)。

Step F5- (3-chloropropyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H,8H, 9H-pyridazino [3, 4-b)]Aza derivatives-9-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

The product of step E (1.35g,2.06mmol,1eq) was dissolved in thionyl chloride (20mL) and stirred at room temperature for 5 h. The reaction solution was concentrated in vacuo, The organic phase was then dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as a yellow gum (989mg,1.47mmol, 71%).

LC/MS(C₃₁H₄₁ClN₆O₃SiS₂)673[M+H]⁺；RT 3.02(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dd,J＝7.8,1.2Hz,1H),7.52-7.39(m,2H),7.30-7.21(m,1H),5.87(s,2H),4.27(q,J＝7.1Hz,2H),4.11-3.98(m,2H),3.72(t,2H),3.67(t,2H),3.19-3.09(m,2H),2.88-2.80(m,2H),2.44(s,3H),2.07-1.96(m,2H),1.94-1.83(m,2H),1.82-1.69(m,2H),1.31(t,J＝7.1Hz,3H),0.91(dd,J＝8.4,7.4Hz,2H),-0.11(s,9H)。

Step G5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H,8H, 9H-pyridazino [3, 4-b)]Aza derivatives-9-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of preparation 4b (369mg,1.91mmol,1.3eq) in dimethylformamide (150mL) was added sodium hydride (147mg,3.67mmol,2.5eq) and the mixture was stirred for 2 min. A solution of the product of step F (989mg,1.47mmol,1eq) in dimethylformamide (50mL) was added and the mixture was heated at 100 ℃ for 1.5 h. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-5% methanol in dichloromethane afforded the desired product as a yellow oil (728mg,0.88mmol, 60%).

LC/MS(C₄₂H₅₂FN₇O₄SiS₂)830[M+H]⁺；RT 2.61(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dd,J＝7.7,1.1Hz,1H),7.52-7.39(m,2H),7.31-7.22(m,2H),7.21-7.15(m,1H),7.12(t,J＝8.7Hz,1H),5.88(s,2H),4.24(q,J＝7.1Hz,2H),4.10(t,2H),4.08-3.99(m,2H),3.77-3.68(m,2H),3.38(s,2H),3.18(t,J＝7.6Hz,2H),2.86-2.77(m,2H),2.43(s,3H),2.20(s,6H),2.11-1.98(m,2H),1.94-1.83(m,2H),1.82-1.69(m,2H),1.27(t,J＝7.1Hz,3H),0.91(dd,J＝8.4,7.4Hz,2H),-0.12(s,9H)。

Step H2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step G (728mg,0.88mmol,1eq) in tetrahydrofuran (18mL) was added ethylenediamine (176. mu.L, 2.63mmol,3eq) and tetrabutylammonium fluoride (1M solution in tetrahydrofuran; 2.64mL,2.63mmol,3eq) and the mixture was heated at 60 ℃ for 24 h. The reaction was partitioned between ethyl acetate and water, the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-7% methanol in dichloromethane afforded the desired product as a yellow gum (224mg,0.32mmol, 37%).

LC/MS(C₃₆H₃₈FN₇O₃S₂)700[M+H]⁺；RT 2.05(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.67(br s,1H),7.87(d,1H),7.51(d,J＝8.0Hz,1H),7.38(ddd,J＝8.2,7.3,1.3Hz,1H),7.26(dd,J＝12.0,2.0Hz,1H),7.23-7.16(m,2H),7.12(t,J＝8.7Hz,1H),4.24(q,J＝7.1Hz,2H),4.09(t,2H),4.05-3.97(m,2H),3.38(s,2H),3.17(t,J＝7.6Hz,2H),2.86-2.77(m,2H),2.40(s,3H),2.20(s,6H),2.11-1.96(m,2H),1.92-1.82(m,2H),1.80-1.69(m,2H),1.27(t,J＝7.1Hz,3H)。

Step I2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H,8H, 9H-pyridazino [3,4-b]Aza derivatives-9-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step H (224mg,0.32mmol,1eq) in 1, 4-dioxane (15mL) was added lithium hydroxide monohydrate (134mg,3.2mmol,10eq) and the mixture was heated at reflux overnight. The reaction was concentrated in vacuo, the residue triturated with water, filtered and dried in vacuo to afford the desired product as a yellow solid (202mg,0.3mmol, 94%) as the lithium salt.

HRMS-ESI(m/z)[M+H]+C₃₄H₃₅FN₇O₃S₂672.2227, found 672.225.

Example 32:2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [3- (methylamino) prop-1-yn-1-yl)]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- {3- [4- (3- { [ (tert-butoxy) carbonyl](methyl) amino } prop-1-yn-1-yl) -2-fluorophenoxy]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of preparation 3g of the product (500mg,0.78mmol,1eq) in toluene (15mL) was added the product of preparation 4c (327mg,1.17mmol,1.5eq), followed by triphenylphosphine (307mg,1.17mmol,1.5eq) and diisopropyl azodicarboxylate (230. mu.L, 1.17mmol,1.5eq) and the mixture was heated to reflux overnight. The reaction was partitioned between dichloromethane and water, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to give the desired product as an off-white foam (715mg,0.79 mmo) l,>100％)。

LC/MS(C₄₅H₅₆FN₇O₆SiS₂)902[M+H]⁺；RT 1.46(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.82(dt,J＝7.6,0.9Hz,1H),7.48-7.37(m,2H),7.33(d,J＝11.6Hz,1H),7.28-7.13(m,3H),5.84(s,2H),4.32-4.17(m,6H),4.15(t,J＝6.1Hz,2H),3.72(dd,J＝8.5,7.4Hz,2H),3.27(d,J＝15.4Hz,2H),2.93-2.75(m,5H),2.36(s,3H),2.19-2.10(m,2H),2.10-1.98(m,2H),1.40(s,9H),1.28(t,3H),0.96-0.89(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [3- (methylamino) prop-1-yn-1-yl-)]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (1.67g,1.85mmol,1eq) in acetonitrile (17mL) was added hydrogen fluoride-pyridine (3.22mL,37mmol,20eq) and the mixture was heated at 60 ℃ for 2 h. The reaction was partitioned between 3:1 dichloromethane/isopropanol and 2N aqueous sodium hydroxide, the organic phase was washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-7% methanol in dichloromethane afforded the desired product as a yellow solid (1.02g,1.52mmol, 82%).

LC/MS(C₃₄H₃₄FN₇O₃S₂)672[M+H]⁺；RT 2.06(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.89(dd,J＝7.8,1.2Hz,1H),7.50(d,J＝8.1Hz,1H),7.38(ddd,J＝8.2,7.3,1.2Hz,1H),7.32-7.25(m,1H),7.23-7.12(m,3H),4.32-4.21(m,4H),4.15(t,J＝6.1Hz,2H),3.45(s,2H),3.32-3.23(m,2H),2.89(t,J＝6.4Hz,2H),2.35(s,3H),2.31(s,3H),2.20-2.10(m,2H),2.09-1.97(m,2H),1.30(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [3- (methylamino) prop-1-yn-1-yl)]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (1.02g,1.52mmol,1eq) in 1, 4-dioxane (50mL) was added lithium hydroxide monohydrate (637mg,15.2mmol,10eq) and the mixture was heated at 110 ℃ overnight. By automatic flash column chromatography (CombiFlash Rf,80g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-70% 0.7N methanolic ammonia in dichloromethane afforded a solid which was triturated with acetonitrile, filtered and dried in vacuo to afford the desired product as a yellow solid (657mg,1.02mmol, 67%).

HRMS-ESI(m/z)[M+H]+C₃₂H₃₁FN₇O₃S₂644.1914, found 644.1930.

Example 332- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-6-hydroxy-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl radicals]-1, 3-thiazole-4-carboxylic acid

Step A(4S) -3- [2- (benzyloxy) acetyl group]-4- (prop-2-yl) -1, 3-oxazolidin-2-one

To a solution of (S) -4-isopropyl-2-oxazolidinone (10g,77.4mmol,1eq) in tetrahydrofuran (200mL) was slowly added sodium hydride (60% dispersion; 3.72g,92.9mmol,1.2eq) at 0 ℃. After 1 h benzyloxyacetyl chloride (12.8mL,81.3mmol,1.05eq) was added dropwise and the mixture was stirred for 1 h. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride (20mL) at 0 ℃ and extracted with ethyl acetate (250 mL). The organic extracts were washed sequentially with water (250mL) and brine (2 × 250mL), dried (magnesium sulfate) and concentrated in vacuo. The solid was suspended in heptane (250mL) and stirred vigorously for 1 h, then filtered, washed with heptane (2X100mL) and dried in vacuo to give the desired product as a white powder (19.9g,71.7mmol, 93%).

LC/MS(C₁₅H₁₉NO₄)278[M+H]⁺；RT 1.19(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.42-7.35(m,4H),7.34-7.28(m,1H),4.64(d,2H),4.57(d,J＝0.8Hz,2H),4.42-4.29(m,3H),2.26-2.15(m,1H),0.85(dd,J＝20.8,6.9Hz,6H)。

Step B(4S) -3- [ (2R) -2- (benzyloxy) hex-4-ynoyl]-4- (prop-2-yl) -1, 3-oxazolidin-2-one

To a solution of the product of step A (8.7g,31.4mmol,1eq) cooled to-78 deg.C in tetrahydrofuran (350mL) was added dropwise sodium bis (trimethylsilyl) amide (1M in tetrahydrofuran; 47.1mL,47.1mmol,1.5eq) and the mixture was stirred at this temperature for 1 hour. A solution of 1-iodobut-2-yne (16.9g,94.1mmol,3eq) in tetrahydrofuran (30mL) was added dropwise and the mixture was warmed to-40 ℃ and stirred for 3 h. The reaction was quenched with saturated aqueous ammonium chloride (200mL), partitioned between ethyl acetate and water, the organic phase was washed successively with water and brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-25% ethyl acetate in isoheptane to give the desired product as a yellow gum (4.17g,12.7mmol, 40%).

LC/MS(C₁₉H₂₃NO₄)330[M+H]⁺；RT 1.31(LCMS-V-B1PsNeg)

¹H NMR(400MHz,DMSO-d6)δ7.41-7.34(m,4H),7.34-7.27(m,1H),5.13(t,J＝5.7Hz,1H),4.57(d,J＝11.9Hz,1H),4.55-4.40(m,2H),4.39-4.26(m,2H),2.68-2.56(m,2H),2.20-2.07(m,1H),1.71(t,J＝2.6Hz,3H),0.84(dd,6H)。

Step C(2R) -2- (benzyloxy) hex-4-yn-1-ol

To a solution of the product of cooling step B (4.17g,12.7mmol,1eq) in tetrahydrofuran (45mL) was added a solution of sodium borohydride (623mg,16.5mmol,1.3eq) in water (12mL) and the mixture was stirred at room temperature for 3 h. The reaction was quenched with saturated aqueous ammonium chloride (100mL) and partitioned between ethyl acetate and water. The organic phase was washed successively with water and brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a colorless oil (2.01g,9.84mmol, 78%).

¹H NMR(400MHz,DMSO-d6)δ7.41-7.31(m,4H),7.31-7.23(m,1H),4.70(t,J＝5.5Hz,1H),4.59(d,J＝1.9Hz,2H),3.57-3.40(m,3H),2.46-2.27(m,2H),1.75(t,J＝2.6Hz,3H)。

Step D{ [ (2R) -2- (benzyloxy) hex-4-yn-1-yl]Oxy } (tert-butyl) diphenylsilane

To a cooled solution of the product of step C (2.01g,9.84mmol,1eq) in dichloromethane (50mL) was added imidazole (1.34g,19.7mmol,2eq) and tert-butyl (chloro) diphenylsilane (5.12mL,19.7mmol,2eq) dropwise and the mixture was warmed to room temperature and stirred for 4 hours. The reaction was quenched with 2M aqueous ammonium chloride and partitioned between dichloromethane and water. The organic phase was washed successively with water and brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-8% ethyl acetate in isoheptane to give the desired product as a colorless oil (3.66g,8.27mmol, 84%).

LC/MS(C₂₉H₃₄O₂Si) weak ionization; RT 1.73(LCMS-V-B1P0sNeg)

¹H NMR(400MHz,DMSO-d6)δ7.68-7.61(m,4H),7.51-7.38(m,6H),7.37-7.32(m,4H),7.32-7.25(m,1H),4.65-4.52(m,2H),3.83-3.68(m,2H),3.67-3.56(m,1H),2.49-2.41(m,2H),1.71(t,J＝2.5Hz,3H),1.00(s,9H)。

Step E4- [ (2R) -2- (benzyloxy) -3- [ (tert-butyldiphenylsilyl) oxy]Propyl radical]-3, 6-dichloro-5-methylpyridazine

A solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (4.99g,33.1mmol,4eq) and the product of step D (3.66g,8.27mmol,1eq) in toluene (50mL) was heated in a sealed flask at 150 ℃ overnight. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,80g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-15% ethyl acetate in isoheptane to give the desired product as a bright orange oil (3.76g,6.65mmol, 80%).

LC/MS(C₃₁H₃₄Cl₂N₂O₂Si)566[M+H]⁺；RT 1.72(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.72-7.62(m,4H),7.56-7.40(m,6H),7.27-7.11(m,3H),6.93-6.84(m,2H),4.44(d,J＝12.1Hz,1H),4.10(d,J＝12.1Hz,1H),3.91-3.78(m,2H),3.76-3.66(m,1H),3.15(dd,J＝13.8,10.0Hz,1H),3.00(dd,J＝13.8,3.6Hz,1H),2.27(s,3H),1.04(s,9H)。

Step F(2R) -2- (benzyloxy) -3- (3, 6-dichloro-5-methylpyridazin-4-yl) propan-1-ol

To a solution of the product of step E (3.76g,6.65mmol,1eq) in tetrahydrofuran (25mL) was added tetrabutylammonium fluoride (1M in tetrahydrofuran; 7.31mL,7.31mmol,1.1eq) and the mixture was stirred at room temperature for 1 hour. The reaction was diluted with ethyl acetate (100mL), washed successively with water (150mL), saturated aqueous sodium bicarbonate (150mL) and brine (2 × 100mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-80% ethyl acetate in isoheptane to afford the desired product as an orange solid (1.68g,5.14mmol, 77%).

LC/MS(C₁₅H₁₆Cl₂N₂O₂)327[M+H]⁺；RT 1.06(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.25-7.12(m,3H),7.01-6.93(m,2H),4.99(t,J＝5.5Hz,1H),4.53(d,J＝12.1Hz,1H),4.22(d,J＝12.0Hz,1H),3.69-3.51(m,3H),3.08-2.91(m,2H),2.29(s,3H)。

Step G2- { [ (2R) -2- (benzyloxy) -3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl]Amino } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step F (1.68g,5.14mmol,1eq) in tetrahydrofuran (55mL) was added triphenylphosphine (2.7g,10.3mmol,2eq) and 2- [ (tert-butoxycarbonyl) amino]Ethyl 1, 3-thiazole-4-carboxylate (1.68g,6.16mmol,1.2eq), then diisopropyl azodicarboxylate (2.02mL,10.3mmol,2eq) was added and the mixture was stirred at room temperature for 2 hours. The reaction was partitioned between ethyl acetate and water, the organic phase was washed successively with water and brine (2 × 50mL), dried (magnesium sulfate) and concentrated in vacuo. The product was dissolved in dichloromethane (30mL), trifluoroacetic acid (7.87mL,103mmol,20eq) was added and the mixture was stirred at room temperature overnight. The reaction solution was diluted and neutralized with a saturated aqueous sodium bicarbonate solution, The layers were separated and the aqueous phase was extracted with dichloromethane (3x100 mL). The combined organic extracts were washed with brine (150mL), dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-80% ethyl acetate in isoheptane to afford the desired product as a yellow solid (1.94g,4.02mmol, 78%).

LC/MS(C₂₁H₂₂Cl₂N₄O₃S)482[M+H]⁺；RT 1.28(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.14(t,J＝5.9Hz,1H),7.57(s,1H),7.24-7.06(m,3H),7.01-6.93(m,2H),4.54(d,J＝11.9Hz,1H),4.31-4.15(m,3H),3.97-3.86(m,1H),3.70-3.59(m,1H),3.52-3.41(m,1H),3.06(dd,J＝13.9,10.2Hz,1H),2.92(dd,J＝13.9,3.2Hz,1H),2.24(s,3H),1.27(t,J＝7.1Hz,3H)。

Step H2- [ (6R) -6- (benzyloxy) -3-chloro-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step G (2.09G,4.34mmol,1eq) in acetonitrile (130mL) was added potassium carbonate (1.2G,8.68mmol,2eq) and copper (I) iodide (827mg,4.34mmol,1eq) and the mixture was heated at reflux for 10 h. The reaction was diluted with water and extracted with ethyl acetate (3 × 60 mL). The combined organic extracts were washed with brine (2 × 50mL), dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automated flash chromatography (CombiFlash Rf, C18130 g redissep column) eluting with a gradient of 5-95% acetonitrile in water afforded the desired product as a light brown solid (0.7g,1.58mmol, 36%).

LC/MS(C₂₁H₂₁ClN₄O₃S)446[M+H]⁺；RT 1.36(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ8.05(s,1H),7.32-7.17(m,5H),4.99-4.86(m,1H),4.59(s,2H),4.39-4.23(m,3H),4.07(dd,J＝13.6,2.1Hz,1H),3.29-3.02(m,2H),2.32(s,3H),1.32(t,J＝7.1Hz,3H)。

Step I2- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-6- (benzyloxy) -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]Pyridazin-8-yl radicals]-1, 3-thiazoles-4-carboxylic acid ethyl ester

To an oven dried, sealed flask was added the product of step H (700mg,1.57mmol,1eq), 2-aminobenzothiazole (473mg,3.15mmol,2eq), N-diisopropylethylamine (0.82mL,4.72mmol,3eq), JosiPhos Pd G3(291mg,0.31mmol,0.2eq) and 1, 4-dioxane (17.5mL) and the mixture was purged with nitrogen (10min) and then heated at 100 ℃ for 3 days. The reaction was partitioned between saturated aqueous sodium bicarbonate and ethyl acetate, the aqueous phase was extracted with ethyl acetate (3 × 80mL), the combined organic extracts were washed with brine (70mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to afford the desired product as a yellow solid (548mg,0.98mmol, 62%).

LC/MS(C₂₈H₂₆N₆O₃S₂)559[M+H]⁺；RT 1.03(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ8.00(s,1H),7.89(d,J＝7.8Hz,1H),7.50(br s,1H),7.38(td,J＝7.7,1.2Hz,1H),7.33-7.16(m,6H),4.96-4.85(m,1H),4.61(s,2H),4.38-4.25(m,3H),4.15-4.05(m,1H),3.25-3.14(m,1H),3.13-3.02(m,1H),2.35(s,3H),1.33(t,J＝7.1Hz,3H)。

Step J2- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-6-hydroxy-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl radicals]-1, 3-thiazole-4-carboxylic acid ethyl ester

To a cooled solution of the product of step I (548mg,0.98mmol,1eq) in dichloromethane (30mL) was added boron trichloride (1M in dichloromethane; 4.9mL,4.9mmol,5eq) dropwise and the mixture was stirred at room temperature for 2 h. The reaction was cooled to 0-5 ℃ and quenched with methanol (5 mL). Methanol (5mL) was added and the mixture heated to reflux for 1.5 h. The reaction was concentrated in vacuo and purified by reverse phase automated flash chromatography (CombiFlash Rf, C1843 g redissep column) eluting with a gradient of 5-95% acetonitrile in water to afford the desired product as a yellow solid (328mg,0.7mmol, 71%).

LC/MS(C₂₁H₂₀N₆O₃S₂)469[M+H]⁺；RT 0.71(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ8.01(d,J＝9.3Hz,1H),7.89(br s,1H),7.51(br s,1H),7.38(t,J＝7.5Hz,1H),7.20(t,J＝7.4Hz,1H),5.38(d,J＝3.2Hz,1H),4.47(dd,J＝12.8,4.8Hz,1H),4.41-4.34(m,1H),4.30(q,J＝7.1Hz,2H),4.19(dd,J＝12.9,2.5Hz,1H),3.08-2.88(m,2H),2.35(s,3H),1.33(t,J＝7.1Hz,3H)。

Step K2- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-6-hydroxy-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl radicals]-1, 3-thiazole-4-carboxylic acid

To a solution of the product of step J (60mg,0.13mmol,1eq) in 1, 4-dioxane (10mL) was added a solution of lithium hydroxide monohydrate (10.8mg,0.26mmol,2eq) in water (2mL) and the mixture was heated at reflux for 2 h. Purification by reverse phase automatic flash chromatography (CombiFlash Rf, C1813 g redissep column) with a gradient elution of 5-95% acetonitrile in water gives a yellow solid. The solid was dissolved in methanol and then loaded onto a short column of PE-AX (10g) wetted with methanol, washed sequentially with methanol and dichloromethane, eluted with 10% formic acid in dichloromethane and concentrated in vacuo to afford the desired product as a yellow solid (11.4mg,0.03mmol, 20%) as the formate salt.

HRMS-ESI(m/z)[M+H]+C₁₉H₁₇N₆O₃S₂441.0804, found 441.0827.

Example 342- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [3- (methylamino) propyl group]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- {3- [4- (3- { [ (tert-butoxy) carbonyl](methyl) amino } propyl) -2-fluorophenoxy]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy ]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of example 32, step A (447mg,0.5mmol,1eq) in 1, 4-dioxane (11mL) under nitrogen was added platinum (IV) oxide (22.5mg,0.1mmol,0.2 eq). The vessel was evacuated and backfilled with nitrogen (x3) and then evacuated, placed under a hydrogen atmosphere and shaken at room temperature for 24 hours. The reaction was filtered through celite, eluted with ethyl acetate and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and elution with a gradient of isoheptane to 50% ethyl acetate in isoheptane afforded the desired product as a yellow solid (424mg,0.47mmol, 94%).

LC/MS(C₄₅H₆₀FN₇O₆SiS₂)906[M+H]⁺；RT 1.45(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.82(d,1H),7.48-7.38(m,2H),7.28-7.20(m,1H),7.12-7.02(m,2H),6.98-6.91(m,1H),5.84(s,2H),4.33-4.20(m,4H),4.08(t,J＝6.2Hz,2H),3.77-3.67(m,2H),3.27(t,J＝7.6Hz,2H),3.18-3.04(m,2H),2.88(t,J＝6.3Hz,2H),2.72(s,3H),2.45(t,J＝7.3Hz,2H),2.37(s,3H),2.17-1.99(m,4H),1.78-1.62(m,2H),1.29(t,3H),1.24(s,9H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [3- (methylamino) propyl group]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (424mg,0.47mmol,1eq) in acetonitrile (6mL) was added hydrogen fluoride-pyridine (0.81mL,9.36mmol,20eq) and the mixture was heated at 60 ℃ for 3 h. The reaction was partitioned between dichloromethane and 2N aqueous sodium hydroxide, the organic phase was washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMCartridge silica) and gradient eluted with 0-15% methanol in dichloromethane afforded the desired product as a yellow solid (147mg,0.22mmol, 47%).

LC/MS(C₃₄H₃₈FN₇O₃S₂)676[M+H]⁺；RT 2.06(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.87(dd,J＝7.9,1.1Hz,1H),7.53-7.45(m,1H),7.41-7.32(m,1H),7.18(td,J＝7.6,1.2Hz,1H),7.12-7.01(m,2H),6.97-6.89(m,1H),4.34-4.19(m,4H),4.09(t,J＝6.2Hz,2H),3.32-3.21(m,4H),2.88(t,J＝6.4Hz,2H),2.41(t,2H),2.34(s,3H),2.24(s,3H),2.17-1.99(m,4H),1.69-1.57(m,2H),1.30(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [3- (methylamino) propyl group]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (147mg,0.22mmol,1eq) in 1, 4-dioxane (15mL) was added lithium hydroxide monohydrate (91.3mg,2.18mmol,10eq) and the mixture was heated at reflux overnight. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-25% 7N methanolic ammonia in dichloromethane afforded a solid which was triturated with acetonitrile, filtered and dried in vacuo to afford the desired product as a yellow solid (83.8mg,0.13mmol, 60%). HRMS-ESI (M/z) [ M + H ]]+C₃₂H₃₅FN₇O₃S₂648.2227, found 648.2246.

Example 355- {3- [4- (aminomethyl) -2-fluorophenoxy group]Propyl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

Step A5- [3- (4-cyano-2-fluorophenoxy) propyl]-2- (4-methyl-3- { [ (2Z) -3- {2- [2- (trimethylsilyl) ethoxy ]Ethyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of preparation 3g of the product (200mg,0.31mmol,1eq) in toluene (6mL) was added 3-fluoro-4-hydroxybenzonitrile (64.2mg,0.47mmol,1.5eq) followed by triphenylphosphine (123mg,0.47mmol,1.5eq) and diisopropyl azodicarboxylate (92.2. mu.L, 0.47mmol,1.5eq) and the mixture was heated to refluxOvernight. The reaction was partitioned between dichloromethane and water, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-50% ethyl acetate in isoheptane to afford the desired product as a beige solid (207mg,0.27mmol, 87%).

LC/MS(C₃₇H₄₂FN₇O₄SiS₂)760[M+H]⁺；RT 3.00(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.89-7.78(m,2H),7.70-7.63(m,1H),7.49-7.38(m,2H),7.36(t,J＝8.6Hz,1H),7.28-7.19(m,1H),5.85(s,2H),4.33-4.18(m,6H),3.77-3.68(m,2H),3.32-3.23(m,2H),2.88(t,J＝6.2Hz,2H),2.37(s,3H),2.22-2.11(m,2H),2.10-1.99(m,2H),1.28(t,J＝7.1Hz,3H),0.96-0.86(m,2H),-0.11(s,9H)。

Step B5- {3- [4- (aminomethyl) -2-fluorophenoxy group]Propyl } -2- (4-methyl-3- { [ (2Z) -3- {2- [2- (trimethylsilyl) ethoxy group]Ethyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

A solution of the product of step A (336mg,0.44mmol,1eq) in ethyl acetate (60mL) was taken upPro (ThalesNano) (110 ℃,85 bar) hydrogenation. The mixture was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMCartridge silica) and gradient eluted with 0-8% methanol in dichloromethane afforded the desired product as a yellow gum (282mg,0.37mmol, 84%).

LC/MS(C₃₇H₄₆FN₇O₄SiS₂)764[M+H]⁺；RT 1.31(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.84-7.78(m,1H),7.48-7.37(m,2H),7.28-7.13(m,2H),7.12-7.00(m,2H),5.84(s,2H),5.87(s,2H),4.33-4.20(m,4H),4.09(td,J＝6.2,1.9Hz,2H),3.78-3.66(m,2H),3.31-3.21(m,4H),2.86(t,J＝6.3Hz,2H),2.36(s,3H),2.19-1.97(m,4H),1.30(t,J＝7.1Hz,3H),0.95-0.84(m,2H),-0.12(s,9H)。

Step C5- {3- [4- (aminomethyl) -2-fluorophenoxy group]Propyl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step B (94mg,0.12mmol,1eq) in tetrahydrofuran (5mL) were added tetrabutylammonium fluoride (1M solution in tetrahydrofuran, 0.74mL,0.74mmol,6eq) and ethylenediamine (49.3. mu.L, 0.74mmol,6eq) and the mixture was heated at 100 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow solid (29mg,0.05mmol, 37%).

LC/MS(C₃₁H₃₂FN₇O₃S₂)634[M+H]⁺；RT 1.96(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.84(d,J＝7.6Hz,1H),7.46(d,J＝7.9Hz,1H),7.38-7.30(m,1H),7.26-7.00(m,4H),4.32-4.21(m,4H),4.09(t,J＝6.2Hz,2H),3.32-3.21(m,4H),2.87(t,J＝6.5Hz,2H),2.33(s,3H),2.17-1.98(m,4H),1.29(t,J＝7.3Hz,3H)。

Step D5- {3- [4- (aminomethyl) -2-fluorophenoxy group]Propyl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step C (29mg,0.05mmol,1eq) in 1, 4-dioxane (5mL) was added lithium hydroxide monohydrate (19.2mg,0.46mmol,10eq) and the mixture was heated at reflux overnight. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-25% 7N methanolic ammonia in dichloromethane to give a solid which was triturated with ether, filtered and dried in vacuo to give the desired product as a yellow solid (3.0mg,4.89 μmol, 28%).

HRMS-ESI(m/z)[M+H]+C₂₉H₂₉FN₇O₃S₂Calculated value of (2): 606.1757, found 606.1782.

Example 362- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of preparation 3g of the product (1g,1.56mmol,1eq) in toluene (30mL) was added the product of preparation 4b (452mg,2.34mmol,1.5eq), followed by triphenylphosphine (614mg,2.34mmol,1.5eq) and diisopropyl azodicarboxylate (461 uL, 2.34mmol,1.5eq) and the mixture was heated to reflux overnight. The reaction was partitioned between dichloromethane and water, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a brown gum (1.16g,1.42mmol, 91%).

LC/MS(C₄₁H₅₀FN₇O₄SiS₂)816[M+H]⁺；RT 2.70(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.84-7.77(m,1H),7.47-7.37(m,2H),7.31(dd,J＝12.0,1.9Hz,1H),7.27-7.12(m,3H),5.83(s,2H),4.32-4.20(m,4H),4.14(t,J＝6.1Hz,2H),3.76-3.67(m,2H),3.37(s,2H),3.31-3.21(m,2H),2.86(t,J＝6.3Hz,2H),2.36(s,3H),2.19(s,6H),2.17-2.09(m,2H),2.07-2.00(m,2H),1.28(t,J＝7.1Hz,3H),0.90(dd,J＝8.5,7.4Hz,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) acetic acidYl) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (262mg,0.32mmol,1eq) in 1, 4-dioxane (10mL) was added ethylenediamine (21.4. mu.L, 0.32mmol,1eq) and tetrabutylammonium fluoride (1M in tetrahydrofuran, 0.96mL,0.96mmol,3eq) and the mixture was heated at 70 ℃ overnight. The reaction was partitioned between ethyl acetate and water, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-6% methanol in dichloromethane afforded the desired product as a yellow gum (114mg,0.17mmol, 52%).

LC/MS(C₃₅H₃₆FN₇O₃S₂)686[M+H]⁺；RT 2.08(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.43(br s,1H),7.88(d,J＝7.6Hz,1H),7.50(d,J＝8.0Hz,1H),7.42-7.35(m,1H),7.31(dd,1H),7.26-7.12(m,3H),4.32-4.22(m,4H),4.15(t,J＝6.1Hz,2H),3.40(s,2H),3.28(dd,J＝8.6,6.7Hz,2H),2.88(t,J＝6.3Hz,2H),2.35(s,3H),2.21(s,6H),2.20-2.10(m,2H),2.07-2.02(m,2H),1.30(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step B (119mg,0.17mmol,1eq) in ethyl acetate (40mL) was added platinum (IV) oxide (7.88mg,0.03mmol,0.2eq) under a nitrogen atmosphere, the vessel was evacuated and back-filled with nitrogen (x3), then evacuated, placed under a hydrogen atmosphere and shaken at room temperature for 3 days. The reaction was filtered through celite, eluted with ethyl acetate and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMCartridge silica) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow gum (55mg,0.08mmol, 46%).

LC/MS(C₃₅H₄₀FN₇O₃S₂)690[M+H]⁺；RT 2.10(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.44(br s,1H),7.88(dd,J＝7.5,1.1Hz,1H),7.50(d,J＝8.1Hz,1H),7.38(td,J＝7.7,1.3Hz,1H),7.20(td,J＝7.5,1.1Hz,1H),7.12-7.02(m,2H),6.97-6.89(m,1H),4.32-4.21(m,4H),4.09(t,2H),3.33-3.22(m,4H),2.89(t,J＝6.3Hz,2H),2.35(s,3H),2.19-2.11(m,4H),2.09(s,6H),2.07-2.01(m,2H),1.63(p,J＝7.4Hz,2H),1.30(t,J＝7.1Hz,3H)。

Step D2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step C (55mg,0.08mmol,1eq) in 1, 4-dioxane (5mL) was added lithium hydroxide monohydrate (33.5mg,0.8mmol,10eq) and the mixture was heated at reflux overnight. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) gradient eluted with 0-20% 7N methanolic ammonia in dichloromethane afforded a solid which was triturated with acetonitrile, filtered and dried in vacuo to afford the desired product as a yellow solid (32.8mg,0.05mmol, 62%).

HRMS-ESI(m/z)[M+H]+C₃₃H₃₇FN₇O₃S₂662.2383, found 662.2402.

Example 372- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-5, 6-dihydropyrrolo [2,3-c ]]Pyridazin-7-yl radicals]-5- [3- [ 2-fluoro-4- [3- (methylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [ tert-Butoxycarbonyl- [2- (3, 6-dichloro-5-methyl-pyridazin-4-yl) ethyl ] methyl]Amino group]-5- [3- (2-fluoro-4-iodo-phenoxy) propyl ]Thiazole-4-carboxylic acid methyl ester

Using the Mitsunobu general procedure, starting with 5.18g of preparation 1a (step D) (9.6mmol,1.0eq.) as the appropriate carbamate and 2.0g of preparation 2c (9.6mmol,1.0eq.) as the appropriate alcohol, 5.6g of the desired product was obtained (80% yield).

Step B2- [2- (3, 6-dichloro-5-methyl-pyridazin-4-yl) ethylamino]-5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for deprotection with HFIP, starting with 5.65g of the product of step A as the appropriate carbamate, 2.9g of the desired product (60%) are obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.76(t,1H),7.59(dd,1H),7.45(dm,1H),6.97(t,1H),4.02(t,2H),3.71(s,3H),3.48(m,2H),3.13(t,2H),3.1(m,2H),2.44(s,3H),1.99(m,2H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,134.2,125,117.7,82.4,67.9,51.8,41.6,30.4,30.4,23.2,16.8；HRMS-ESI(m/z):[M+H]+C₂₁H₂₁Cl₂FIN₄O₃Calculated value of S is 624.9734, found value is 624.9740.

Step C2- (3-chloro-4-methyl-5, 6-dihydropyrrolo [2, 3-c)]Pyridazin-7-yl) -5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid methyl ester

A suspension of 3.0g of the product of step B (4.79mmol,1eq.) and 1.85g of cesium carbonate (9.59mmol,2eq.) in 25mL of dry 1, 4-dioxane was stirred at 80 ℃ for 3 hours and complete conversion was observed. The reaction mixture was directly evaporated onto celite and then purified by flash chromatography using DCM-MeOH as eluent to give 2.64g of the desired product (93% yield).¹H NMR(400MHz,DMSO-d₆)δppm 7.6(dd,1H),7.46(dm,1H),6.99(t,1H),4.35(t,2H),4.1(t,2H),3.79(s,3H),3.3(m,2H),3.3(m,2H),2.28(s,3H),2.12(m,2H)；¹³C NMR(100MHz,DMSO-d₆)δppm 134.2,125.0,117.7,68.1,52.2,49.8,30.7,24.6,23.5,16.0；HRMS-ESI(m/z):[M+H]+C₂₁H₂₀ClFIN₄O₃Calculated value of S is 588.9967, found value is 588.9959.

Step D5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group ]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-5, 6-dihydropyrrolo [2, 3-c)]Pyridazin-7-yl) thiazole-4-carboxylic acid methyl ester

A100 mL oven-dried single-necked round bottom flask was equipped with a PTFE-coated magnetic stir bar and placed inAnd (4) a reflux condenser is arranged. To this was added 2.6g of the product of step C (4.48mmol,1eq.), 1.51g N-methyl-N-prop-2-ynyl-carbamic acid tert-butyl ester (8.96mmol,2eq.) and 4mL DIPEA, followed by 16mL of dry THF and flushing the system with argon. After stirring for 5 minutes under an inert atmosphere, 42mg of Pd (PPh) was added₃)₂Cl₂(0.224mmol,0.05eq.) and 79mg (0.224mmol,0.05 eq.). The resulting mixture was then warmed to 60 ℃ and stirred at this temperature for 2 hours to achieve complete conversion. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane-EtOAc as eluent to give 1.88g of the desired product (67% yield).

Step E2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-5, 6-dihydropyrrolo [2,3-c ]]Pyridazin-7-yl radicals]-5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A4 mL oven-dried vial was equipped with a PTFE-coated magnetic stir bar to which were added 80mg of the product of step D (0.127mmol,1.0eq.), 28mg of 1, 3-benzothiazol-2-amine (0.19mmol,1.5eq.), and 113uL of DIPEA (0.635mmol, 5eq.) suspended in 0.5mL dry 1, 4-dioxane. The resulting mixture was flushed with nitrogen and 11mg Pd was added ₂(dba)₃(0.012mmol,0.1eq.) and 14mg XantPhos (0.024mmol,0.2 eq.). The reaction mixture was warmed to 120 ℃ and stirred at this temperature for 2 hours at which time the reaction gave complete conversion. The reaction mixture was directly evaporated onto celite and then purified by flash chromatography using heptane-EtOAc as eluent to give 65mg of the desired product (68% yield).

Step F2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-5, 6-dihydropyrrolo [2,3-c ]]Pyridazin-7-yl radicals]-5- [3- [ 2-fluoro-4- [3- (methylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure of deprotection and hydrolysis, starting from the product of step E as the appropriate methyl ester, the desired product is obtained.

HRMS-ESI(m/z):[M+H]+C₃₁H₂₉FN₇O₃S₂630.1752, foundValue 630.1755.

Example 382- [3- (1, 3-benzothiazol-2-ylamino) -6-hydroxy-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (methylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid (enantiomerically pure, enantiomer 2 from preparation 2b, step A)

Step A2- [ tert-butoxycarbonyl- [2- [ tert-butyl (diphenyl) silyl ] group]Oxy-3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propyl]Amino group]-5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group ]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester (enantiomerically pure, enantiomer 2 from preparation 2b, step A)

Using the Mitsunobu general procedure, starting with 1.91g of preparation 1b (3.4mmol,1.0eq.) as the appropriate carbamate and 1.6g of preparation 2b (3.4mmol,1.0eq.) as the appropriate alcohol, 2.2g of the desired product was obtained (63% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.54-7.26(m,10H),7.29(m,1H),7.18(d,1H),7.08(t,1H),4.76(m,1H),4.34/3.84(m,2H),4.06(t,2H),3.75(s,3H),3.23(m,2H),2.96/2.89(m,2H),2.86(bs,3H),2.06(m,2H),2.03(bs,3H),1.4(s,18H),1.22(m,2H),0.81(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 129.0,119.2,115.4,68.8,67.9,52.0,51.1,36.1,33.9,30.4,28.0,26.9,22.9,22.7,19.1,16.8；HRMS-ESI(m/z):[M+H]+C₅₂H₆₃Cl₂FN₅O₈Calculated value of SSi 1034.3522, found value 1034.3519.

Step B5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [ [2- [ tert-butyl (diphenyl) silyl ] group]Oxy-3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propyl]Amino group]Thiazole-4-carboxylic acid methyl ester (enantiomerically pure, enantiomer 2 from preparation 2b, step A)

Using the general procedure for deprotection with HFIPA, starting from the product of step A as the appropriate carbamate, 1.6g of the desired product was obtained (81% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.76(t,1H),7.56-7.26(m,10H),7.31(d,1H),7.21(d,1H),7.12(t,1H),4.46(m,1H),4.23(br.,2H),4.06(t,2H),3.68(s,3H),3.38/3.25(m+m,2H),3.11(t,2H),3.05(m,2H),2.86(br.,3H),2.11(s,3H),1.98(quint.,2H),1.41(s,9H),0.85(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.9,157.4,157.3,154.9,151.5,140.6,139.8,129.1,119.3,115.3,69.9,67.9,51.7,50.1,38.5,36.2,33.8,30.5,28.5,27.1,23.2,17.0；HRMS-ESI(m/z):[M+H]+C₄₇H₅₅Cl₂FN₅O₆Calculated value of SSi 934.2997, found value 934.2994.

Step C5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [6- [ tert-butyl (diphenyl) silyl]Oxy-3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester (enantiomerically pure, enantiomer 2 from preparation 2b, step A)

A40 mL oven dried vial was equipped with a PTFE-coated magnetic stir bar to which was added 1.50g of the product of step B (1.6mmol,1.0eq) dissolved in 8mL 1, 4-dioxane followed by 620mg cesium carbonate (3.2mmol,2.0eq) and 560uL DIPEA (410mg,3.2mmol,2.0eq) and placed under an inert atmosphere. 110mg Pd (AtaPhos) was added₂Cl₂After (0.16mmol,0.10eq.) the reaction mixture was warmed to 80 ℃ and stirred at this temperature for 30 minutes at which time the reaction gave complete conversion. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using heptane and EtOAc as eluent to give 550mg of the desired product (38% yield).

¹H NMR(500MHz,DMSO-d₆)δppm 7.47(dm,4H),7.47/7.43(tm+tm,2H),7.38/7.33(tm+tm,4H),7.3(dm,1H),7.18(m,1H),7.11(t,1H),4.72/3.86(m+m,2H),4.63(m,1H),4.22(s,2H),4.11(t,2H),3.79(s,3H),3.27(m,2H),2.95/2.87(m+m,2H),2.85(s,3H),2.14(s,3H),2.12(m,2H),1.41(s,9H),0.8(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.0,156.0,151.9,151.6,151.5,151.4,147.5,142.2,137.0,135.6/135.5,135.0,133.2/133.1,130.6/130.5,129.1,128.4/128.3,126.8,119.4,115.4,114.8,85.3,82.5,79.8,68.3,63.1,52.0,51.8,38.5,33.8,32.6,30.7,28.5,26.8,23.1,19.1,15.5；HRMS-ESI(m/z):[M+H]+C₄₇H₅₄ClFN₅O₆Calculated value of SSi 898.3231, found value 898.3238.

Step D2- [3- (1, 3-benzothiazol-2-ylamino) -6- [ tert-butyl (diphenyl) silyl]Oxy-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester (enantiomerically pure, enantiomer 2 from preparation 2b, step A)

A 4mL oven dried vial was equipped with a PTFE-coated magnetic stir bar to which were added 179mg of the product of step C (0.2mmol,1.0eq.), 60mg of 1, 3-benzothiazol-2-amine (0.4mmol,2.0eq.), and 104uL of DIPEA (0.6mmol,3eq) dissolved in 1mL dry DMF. The resulting mixture was flushed with nitrogen and 18mg Pd was added ₂(dba)₃(0.02mmol,0.1eq.) and 23mg XantPhos (0.04mmol,0.2 eq.). The reaction mixture was warmed to 140 ℃ and stirred at this temperature for 30 minutes at which time the reaction gave complete conversion. The reaction mixture was injected directly onto a pre-treated silica gel column, which was then purified by flash chromatography using heptane and EtOAc as eluent to give 50mg of the desired product (24% yield).

Step E2- [3- (1, 3-benzothiazol-2-ylamino) -6-hydroxy-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (methylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid (enantiomerically pure, enantiomer 2 from preparation 2b, step A)

Using the general procedure of deprotection and hydrolysis, starting from the product of step D as the appropriate methyl ester, the desired product is obtained.

HRMS-ESI(m/z):[M+H]+C₃₂H₃₁FN₇O₄S₂660.1857, found 660.1847.

Example 392- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-4-A6- { [2- (methylamino) ethyl radical]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A(4R) -4- (2-hydroxyethyl) -2, 2-dimethyl-1, 3-oxazolidine-3-carboxylic acid tert-butyl ester

To N- [ (2R) -1, 4-dihydroxybutan-2-yl]To a solution of tert-butyl carbamate (10.5g,51.2mmol,1eq) in dichloromethane (110mL) was added 2, 2-dimethoxypropane (12.5mL,102mmol,2eq) and p-toluenesulfonic acid monohydrate (0.51mL,5.12mmol,0.1eq) and the mixture was stirred at room temperature overnight. The reaction was quenched with 0.2N aqueous sodium hydroxide (50mL), and the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to afford the desired product as a white solid (5.13g,20.9mmol, 41%).

¹H NMR(400MHz,DMSO-d6)δ4.49-4.40(m,1H),3.92-3.82(m,2H),3.79(d,J＝7.8Hz,1H),3.48-3.39(m,2H),1.87-1.69(m,1H),1.66-1.50(m,1H),1.47(s,3H),1.42(s,12H)。

Step B(4R) -2, 2-dimethyl-4- (2-oxoethyl) -1, 3-oxazolidine-3-carboxylic acid tert-butyl ester

A solution of the product of step A (5.13g,20.9mmol,1eq) in dichloromethane (100mL) was cooled to 0 ℃. Dess-Martin periodinane (9.31g,22mmol,1.05eq) was added and the mixture was stirred at room temperature for 2.5 h. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,220g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a clear oil (3.23g,13.3mmol, 64%).

¹H NMR(400MHz,DMSO-d6)δ9.65(d,J＝6.7Hz,2H),4.22(t,J＝6.0Hz,2H),4.09-3.98(m,1H),3.71(t,J＝9.0Hz,1H),2.76-2.61(m,2H),1.48(s,3H),1.40(d,J＝8.1Hz,12H)。

Step C(4R) -4- (3, 3-dibromoprop-2-en-1-yl) -2, 2-dimethyl-1, 3-oxazolidine-3-carboxylic acid tert-butyl ester

To a solution of tetrabromomethane (4.04mL,39.8mmol,3eq) in dichloromethane (65mL) was added a solution of triphenylphosphine (20.9g,79.7mmol,6eq) in dichloromethane (100mL) and the mixture was stirred for 10 min. To the solution was added a solution of the product of step B (3.23g,13.3mmol,1eq) in dichloromethane (60mL) and triethylamine (16.6mL,120mmol,9eq) and the mixture was cooled to 0 ℃. The first solution was added in portions and the resulting mixture was stirred at 0 ℃ for 2 hours. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate, the organic phase was washed with brine, dried (PTFE phase separator) and concentrated in vacuo. The residue was triturated with ice-cold ether, the solid collected by filtration and washed with ether. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-15% ethyl acetate in isoheptane to give the desired product as a clear oil (1.77g,4.43mmol, 33%).

¹H NMR(400MHz,DMSO-d6)δ6.61(t,J＝7.5Hz,1H),4.08-3.87(m,2H),3.83-3.70(m,2H),2.45-2.28(m,2H),1.50(s,3H),1.41(d,12H)。

Step D(4R) -4- (but-2-yn-1-yl) -2, 2-dimethyl-1, 3-oxazolidine-3-carboxylic acid tert-butyl ester

To a solution of the product of step C (1.77g,4.43mmol,1eq) cooled to-78 deg.C in tetrahydrofuran (40mL) was added n-butyllithium (2.5M in hexane; 5.32mL,13.3mmol,3eq) and the mixture was stirred at this temperature for 1 hour. Methyl iodide (1.38mL,22.2mmol,5eq) was added and the mixture was warmed to room temperature and stirred overnight. The reaction was partitioned between ethyl acetate and saturated aqueous ammonium chloride, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMCartridge silica gel) and gradient eluted with 0-15% ethyl acetate in isoheptane to give the desired product as a clear oil (1.04g,4.11mmol, 93%).

¹H NMR(400MHz,DMSO-d6)δ4.02-3.92(m,1H),3.90-3.75(m,2H),2.46(d,J＝15.8Hz,1H),2.37-2.20(m,1H),1.75(t,J＝2.6Hz,3H),1.48(s,3H),1.41(dd,J＝8.2,3.8Hz,12H)。

Step EN- [ (2R) -1-hydroxyhex-4-yn-2-yl]Carbamic acid tert-butyl ester

To a solution of the product of step D (1.04g,4.11mmol,1eq) in methanol (30mL) was added p-toluenesulfonic acid monohydrate (0.08mL,0.82mmol,0.2eq) and the mixture was stirred at room temperature overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-60% ethyl acetate in isoheptane to give the desired product as a clear oil (724mg,3.39mmol, 83%).

¹H NMR(400MHz,DMSO-d6)δ6.54(d,J＝8.3Hz,1H),4.67(t,J＝5.7Hz,1H),3.53-3.40(m,1H),3.39-3.26(m,2H),2.38-2.09(m,2H),1.73(t,J＝2.6Hz,3H),1.39(s,9H)。

Step FN- [ (2R) -1- [ (tert-butyldiphenylsilyl) oxy]Hex-4-yn-2-yl]Carbamic acid tert-butyl ester

To a solution of the product of step E (724mg,3.39mmol,1eq) in dichloromethane (30mL) was added imidazole (0.45mL,6.79mmol,2eq) and the mixture was cooled to 0 ℃, then tert-butyl (chloro) diphenylsilane (0.93mL,3.56mmol,1.05eq) was added and the mixture was stirred at room temperature overnight. The reaction was partitioned between dichloromethane and saturated aqueous ammonium chloride, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMCartridge silica) and gradient eluted with 0-8% ethyl acetate in isoheptane to give the desired product as a clear oil (1.67g,3.7mmol, 109%).

LC/MS(C₂₇H₃₇NO₃Si)352[M-Boc]⁺；RT 1.28(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.73-7.59(m,4H),7.52-7.35(m,6H),3.67-3.57(m,2H),2.46-2.14(m,2H),1.70(t,J＝2.5Hz,3H),1.39(s,9H),1.37-1.29(m,1H),1.00(s,9H)。

Step G{ [ (2R) -2-aminohex-4-yn-1-yl]Oxy } (tert-butyl) diphenylsilane

To a solution of the product of step F (1.67g,3.7mmol,1eq) cooled to 0 ℃ in dichloromethane (35mL) was added trifluoroacetic acid (8.07mL,105mmol,28.5eq) and the mixture was stirred at room temperature for 1 hour. The reaction was partitioned between dichloromethane and 1N aqueous sodium hydroxide (120mL), the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to give the desired product as a clear oil (997mg,2.84mmol, 77%).

¹H NMR(400MHz,DMSO-d6)δ7.68-7.61(m,4H),7.53-7.40(m,6H),3.54(d,J＝5.7Hz,2H),2.92-2.81(m,1H),2.38-2.14(m,2H),1.73(t,J＝2.6Hz,3H),1.62-1.44(m,2H),1.01(s,9H)。

Step HN- (2- { [ (2R) -1- [ (tert-butyldiphenylsilyl) oxy]Hex-4-yn-2-yl]Amino } ethyl) -N-methylcarbamic acid benzyl ester

To a solution of the product of step G (997mg,2.84mmol,1eq) in tetrahydrofuran (24mL) was added a solution of benzyl N-methyl-N- (2-oxoethyl) carbamate (647mg,3.12mmol,1.1eq) in tetrahydrofuran (11.9mL) and the mixture was cooled to 0 ℃. Sodium triacetoxyborohydride (1.8g,8.51mmol,3eq) was added and the mixture was warmed to room temperature and stirred for 4 h. The reaction was partitioned between ethyl acetate and 1N aqueous sodium hydroxide, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the desired product as a clear oil (1.46g,2.69mmol, 95%).

LC/MS(C₃₃H₄₂N₂O₃Si)543[M+H]⁺；RT 2.34(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.63(d,J＝6.7Hz,4H),7.52-7.39(m,6H),7.37-7.24(m,5H),5.03(d,J＝2.6Hz,2H),3.64-3.52(m,2H),3.28(t,J＝6.5Hz,2H),2.86(d,J＝14.2Hz,3H),2.74-2.60(m,2H),2.35-2.23(m,2H),1.68(t,3H),1.63-1.50(m,1H),0.99(s,9H)。

Step IN- (2- { [ (benzyloxy) carbonyl](methyl) amino } ethyl) -N- [ (2R) -1- [ (tert-butyldiphenylsilyl) oxy]Hex-4-yn-2-yl]Carbamic acid benzyl ester

To the product of step H (1.46g,2.69mmol,1eq) in ethyl acetate (30mL) was added a solution of sodium bicarbonate (497mg,5.92mmol,2.2eq) in water (8.5mL) and the mixture was cooled to 0 deg.C, then benzyl chloroformate (0.46mL,3.23mmol,1.2eq) was added and the mixture was warmed to room temperature and stirred overnight. The reaction was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a clear oil (1.75g,2.59mmol, 96%).

LC/MS(C₄₁H₄₈N₂O₅Si)677[M+H]⁺；RT 1.32(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.67-7.50(m,4H),7.49-7.22(m,16H),5.16-4.90(m,4H),4.19-4.07(m,1H),3.79-3.51(m,2H),3.44-3.34(m,2H),3.28-3.15(m,2H),2.92-2.71(m,3H),2.45-2.17(m,2H),1.64(s,3H),0.95(d,J＝4.4Hz,9H)。

Step JN- (2- { [ (benzyloxy) carbonyl](methyl) amino } ethyl) -N- [ (2R) -1- [ (tert-butyldiphenylsilyl) oxy]-3- (3, 6-dichloro-5-methylpyridazin-4-yl) propan-2-yl]Carbamic acid benzyl ester

To a solution of the product of step I (1.75g,2.59mmol,1eq) in toluene (35mL) was added 3, 6-dichloro-1, 2,4, 5-tetrazine (1.56g,10.3mmol,4eq) and the mixture was heated in a sealed flask at 150 ℃ for 3 days. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMCartridge silica) and gradient eluted with 0-40% ethyl acetate in isoheptane to give the crude desired product as a red gum (1.2g,1.5mmol, 58%) which was used directly in the next step without further purification.

LC/MS(C₄₃H₄₈N₄O₅SiCl₂)799[M+H]⁺；RT 1.32(LCMS-V-B2)

Step KN- (2- { [ (benzyloxy) carbonyl](methyl) amino } ethyl) -N- [ (2R) -1- (3, 6-dichloro-5-methylpyridazin-4-yl) -3-hydroxypropan-2-yl]Benzyl carbamate

To a solution of the product of step J (1.2g,1.5mmol,1eq) in methanol (55mL) was added acetyl chloride (0.54mL,7.5mmol,5eq) and the mixture was stirred at room temperature for 3 days. The reaction was concentrated in vacuo, partitioned between dichloromethane and water, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the crude desired product as an orange gum (846mg,1.51mmol, 100%) which was used directly in the next step without further purification.

LC/MS(C₂₇H₃₀Cl₂N₄O₅)561[M+H]⁺；RT 0.80(LCMS-V-B2)

Step L2- { [ (tert-butoxy) carbonyl]Amino } -5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 4d (652mg,3.31mmol,1.2eq) in dimethylformamide (15mL) cooled to 0 deg.C was added sodium hydride (60% dispersion; 264mg,6.61mmol,2.4eq) and the mixture was stirred for 30 min. A solution of the product of preparation 1d (1.17g,2.75mmol,1eq) in dimethylformamide (15mL) was added and the mixture was stirred at room temperature for 1 h. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMCartridge silica) and gradient eluted with 0-16% methanol in dichloromethane afforded the desired product as a clear gum (431mg,0.87mmol, 32%).

LC/MS(C₂₄H₃₄FN₃O₅S)496[M+H]⁺；RT 1.81(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.64(br s,1H),7.13-7.00(m,2H),6.97-6.89(m,1H),4.04(t,J＝6.2Hz,2H),3.75(s,3H),3.28-3.15(m,2H),2.58-2.50(m,2H),2.18(t,J＝7.2Hz,2H),2.12(s,6H),2.12-2.00(m,2H),1.71-1.59(m,2H),1.47(s,9H)。

Step M2- { [ (2R) -2- { [ (benzyloxy) carbonyl](2- { [ (benzyloxy) carbonyl](methyl) amino } ethyl) amino } -3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl][ (tert-butoxy) carbonyl]Amino } -5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step K (422mg,0.75mmol,1eq) in toluene (25mL) was added the product of step L (431mg,0.87mmol,1.16eq), followed by triphenylphosphine (394mg,1.5mmol,2eq) and di-tert-butyl azodicarboxylate (346mg,1.5mmol,2eq) and the mixture was stirred at room temperature for 3 hours. The reaction was partitioned between dichloromethane and water, the organic phase was washed with brine, separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as an orange gum (517mg,0.5mmol, 66%).

LC/MS(C₅₁H₆₂Cl₂FN₇O₉S)1040[M+H]⁺；RT 2.48(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.31(s,27H),7.24(s,2H),7.11-6.95(m,8H),6.90(d,J＝8.6Hz,3H),5.76(s,3H),5.22(s,1H),5.05(s,1H),4.97(s,4H),4.78(s,1H),4.50(d,J＝12.9Hz,1H),4.00(s,7H),3.69(s,6H),2.94(s,3H),2.82(s,3H),2.76(s,1H),2.37(s,1H),2.30(s,1H),2.20(s,7H),2.13(s,17H),2.09(s,1H),2.05(s,3H),1.96(s,5H),1.64(d,J＝8.1Hz,8H),1.50(s,5H),1.43(d,J＝10.5Hz,6H)。

Step N2- { [ (2R) -2- { [ (benzyloxy) carbonyl](2- { [ (benzyloxy) carbonyl](methyl) amino } ethyl) amino } -3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl ]Amino } -5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step M (517mg,0.5mmol,1eq) in dichloromethane (10mL) was added trifluoroacetic acid (4mL,52.2mmol,105eq) andthe mixture was stirred at room temperature for 1 hour. The reaction was diluted with dichloromethane and cooled to 0 ℃, the reaction was quenched with 1N aqueous sodium hydroxide (60mL) and the organic phase was washed with brine, separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-8% 1N methanolic ammonia in dichloromethane afforded the desired product as an orange gum (461mg,0.49mmol, 99%).

LC/MS(C₄₆H₅₄Cl₂FN₇O₇S)939[M+H]⁺；RT 2.16(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.88(s,1H),7.48-7.18(m,10H),7.11-6.88(m,3H),5.12-4.82(m,4H),4.06-3.90(m,2H),3.76-3.58(m,4H),3.53-3.36(m,2H),3.29-3.21(m,2H),3.17-3.03(m,4H),3.01-2.81(m,3H),2.79-2.69(m,2H),2.36-2.24(m,4H),2.18(t,2H),2.11(s,6H),2.00-1.93(m,3H),1.64(p,J＝7.3Hz,2H)。

Step O2- [ (6R) -6- { [ (benzyloxy) carbonyl](2- { [ (benzyloxy) carbonyl](methyl) amino } ethyl) amino } -3-chloro-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl radicals]-5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step N (461mg,0.49mmol,1eq) in 1, 4-dioxane (10mL) was added cesium carbonate (320mg,0.98mmol,2eq) and N, N-diisopropylethylamine (171. mu.L, 0.98mmol,2 eq). The mixture was purged with nitrogen (10min), then bis (di-tert-butyl (4-dimethylaminophenyl) phosphine) palladium (II) dichloride (34.8mg,0.05mmol,0.1eq) was added and the mixture was heated at 80 ℃ for 3 h under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-12% methanol in dichloromethane afforded the desired product as an orange gum (285mg,0.32mmol, 64%).

LC/MS(C₄₆H₅₃ClFN₇O₇S)902[M+H]⁺；RT 2.32(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.52-7.21(m,9H),7.15-7.00(m,4H),5.21-4.91(m,4H),4.84-4.63(m,1H),4.39-4.22(m,1H),4.07(t,J＝6.2Hz,2H),4.05-3.91(m,1H),3.77(s,3H),3.56-3.43(m,2H),3.41-3.36(m,2H),3.31(s,2H),3.30-3.23(m,2H),3.01-2.75(m,4H),2.28(s,2H),2.18(d,J＝6.9Hz,4H),2.11(s,6H),1.64(p,J＝7.4Hz,2H)。

Step P2- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-6- { [ (benzyloxy) carbonyl](2- { [ (benzyloxy) carbonyl](methyl) amino } ethyl) amino } -4-methyl-5H, 6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step O (117mg,0.13mmol,1eq) in 1, 4-dioxane (5mL) were added 2-aminobenzothiazole (39mg,0.26mmol,2eq), N-diisopropylethylamine (67.9. mu.L, 0.39mmol,3eq) and Josiphos Pd G3(24mg,0.03mmol,0.2eq) and the mixture was purged with nitrogen (10min) and then heated at 140 ℃ for 3 hours under microwave irradiation. The reaction was partitioned between ethyl acetate and water, the organic phase washed with brine, dried (magnesium sulfate) and concentrated in vacuo. Purification by reverse phase automatic flash chromatography (CombiFlash Rf, C1813 g redissep column) eluting with a gradient of 5-95% acetonitrile in water afforded the crude desired product as a yellow gum (23mg,0.02mmol, 17%) which was used directly in the subsequent step without further purification.

LC/MS(C₅₃H₅₈FN₉O₇S₂)1017[M+H]⁺；RT 2.43(LCMS-V-C)

Step Q2- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6- { [2- (methylamino) ethyl]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- (3- {4- [3- (dimethylamino) propyl group]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step P (23mg,0.02mmol,1eq) in 1, 4-dioxane (0.5mL) was added concentrated hydrochloric acid (1.5mL) and the mixture was stirred at room temperature for 3 days. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% 7N methanolic ammonia in dichloromethaneA solid was obtained which was triturated with ether, filtered and dried in vacuo to give the desired solid as a yellow solid (9.5mg,0.01mmol, 57%).

HRMS-ESI(m/z)[M+H]+C₃₆H₄₅FN₉O₃S₂734.3071, found 734.3096.

Example 402- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2- (trifluoromethyl) phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- {3- [ 4-bromo-2- (trifluoromethyl) phenoxy group]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene ]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

Diisopropyl azodicarboxylate (0.09mL,0.47mmol,1.5eq) was added dropwise to a solution of preparation 3g of the product (200mg,0.31mmol,1eq), 4-bromo-2- (trifluoromethyl) phenol (100mg,0.41mmol,1.33eq) and triphenylphosphine (123mg,0.47mmol,1.5eq) in toluene (10 mL). The mixture was stirred at room temperature for 18 hours, then concentrated in vacuo. Purification by flash chromatography (20g silica gel) eluting with a gradient of 0-25% ethyl acetate in isoheptane afforded the desired product as a yellow solid (265mg,0.26mmol, 84%).

LC/MS(C₃₇H₄₂BrF₃N₆O₄SiS₂)863[M+H]⁺；RT 1.91(LCMS-V-B1)

¹H NMR(400MHz,CDCl₃)δ7.71-7.64(m,1H),7.62-7.52(m,2H),7.39-7.34(m,2H),7.24-7.17(m,1H),6.88(d,J＝8.9Hz,1H),5.84(s,2H),4.50-4.30(m,4H),4.12(t,J＝6.3Hz,2H),3.82-3.68(m,2H),3.39-3.30(m,2H),2.87(t,J＝6.4Hz,2H),2.38(s,3H),2.33-2.22(m,2H),2.18-2.07(m,2H),1.40(t,J＝7.1Hz,3H),1.01-0.90(m,2H),-0.08(s,9H)。

Step B5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2- (trifluoromethyl) phenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (265mg,0.26mmol,1eq) in 2-methyltetrahydrofuran (10mL) was added copper (I) iodide (9.93mg,0.05mmol,0.2eq) and tetrakis (triphenylphosphine) palladium (0) (30.1mg,0.03mmol,0.1eq) and dimethyl (prop-2-yn-1-yl) amine (0.1mL,0.93mmol,3.5 eq). N, N-diisopropylethylamine (0.14ml,0.78mmol,3eq) was added and the mixture was heated at 75 ℃ for 96 h. The reaction was cooled to room temperature and concentrated in vacuo. Purification by flash chromatography (20g of silica gel) eluting with a gradient of 0-100% ethyl acetate in isoheptane afforded the desired product as a dark yellow gum (170mg,0.2mmol, 75%).

LC/MS(C₄₂H₅₀F₃N₇O₄SiS₂)866[M+H]⁺；RT 1.57(LCMS-V-B1)

¹H NMR(400MHz,CDCl₃)δ7.66(d,J＝2.3Hz,1H),7.59(dt,J＝7.7,0.9Hz,1H),7.54-7.50(m,1H),7.40-7.31(m,2H),7.23-7.17(m,1H),6.92(d,J＝8.6Hz,1H),5.84(s,2H),4.49-4.32(m,4H),4.18-4.12(m,2H),3.80-3.66(m,2H),3.42(s,2H),3.35(dd,J＝8.5,6.6Hz,2H),2.87(t,J＝6.4Hz,2H),2.38(s,3H),2.34(s,6H),2.31-2.25(m,2H),2.17-2.08(m,2H),1.40(t,J＝7.1Hz,3H),0.99-0.90(m,2H),-0.07(s,9H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2- (trifluoromethyl) phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (160mg,0.18mmol,1eq) in 1, 4-dioxane (4.5mL) at 0 deg.C was added trifluoroacetic acid (0.5mL,6.47mmol,35eq) and the mixture was stirred for 36 h. The reaction was diluted with dichloromethane (50mL) and washed sequentially with 1M aqueous ammonia (20mL), water (20mL) and brine (20mL), dried (magnesium sulfate) and concentrated in vacuo. To a suspension of the crude product in a mixture of water (1.5mL) and tetrahydrofuran (1.5mL) was added lithium hydroxide monohydrate (23.3mg,0.55mmol,3eq) and the mixture was stirred at room temperature for 72 h. Water (5mL) was added and the suspension was neutralized with acetic acid. The solid was collected by filtration, washed with water (10mL) and dried in vacuo to give the desired product as a yellow solid (75mg,0.11mmol, 57%).

HRMS-ESI(m/z)[M+H]+C₃₄H₃₃F₃N₇O₃S₂708.2038, found 708.2058.

EXAMPLE 412- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A PTFE-coated magnetic stir bar was assembled in a 24mL oven-dried vial and 250mg of 1-methylpiperazine (2.5mmol,5.0eq.) dissolved in 2.5mL dry THF was added to it. 133mg of 3-bromobut-1-yne (1.0mmol,2.0eq) are then added dropwise via syringe over 5 minutes and stirred at this temperature for 30 minutes. To the resulting mixture was added 301mg of preparation 3a (0.50mmol,1.0eq.), 18.15mg Pd (PPh)₃)₂Cl₂(0.025mmol,0.05eq.) and 4.76CuI (0.025mmol,0.05eq.), then heated to 60 ℃ and stirred at that temperature for 2 hours. The reaction was completely converted. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using DCM and MeOH (1.2% NH₃) As eluent 300mg (95% yield) of the desired product was obtained.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-Carboxylic acid methyl ester

Using Buchwald general procedure II, starting with 300mg of the product of step A (0.47mmol,1.0eq.) and 140mg of 1, 3-benzothiazol-2-amine (0.94mmol,2.0eq.) gives 150mg (42%) of the desired product.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure for hydrolysis, the starting material used the product of step B as the appropriate methyl ester to give the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.87(d,1H),7.49(d,1H),7.36(t,1H),7.26(dd,1H),7.2(t,1H),7.16(dd,1H),7.13(t,1H),4.27(t,2H),4.12(t,2H),3.65(q,1H),3.27(t,2H),2.87(t,2H),2.62-2.21(brm,8H),2.14(s,3H),2.13(qn,2H),2.04(qn,2H),1.33(s,3H),1.25(d,3H)；¹³C NMR(125MHz,DMSO-d₆)δppm 164.3,155.4,151.5,151.4,148.6,147.2,145.1,140.2,136.3,130.2,129.0,129.0,127.6,126.5,122.5,122.3,119.2,116.4,115.5,115.4,88.4,84.1,68.5,51.7,46.3,46.1,31,23.9,23.0,20.3,19.6,12.9；

HRMS-ESI(m/z)[M+H]⁺C₃₇H₄₀FN₈O₃S₂Calculated value of (2) 727.2649, found value of 727.2630

Example 422- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using propynylamine preparation general procedure using 258mg of preparation 3d (0.40mmol,1eq.) as the appropriate propargyl alcohol and pyrrolidine (20eq,670mg) gave 120mg of the desired product (43%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure for hydrolysis, the starting material uses the product of step a as the appropriate methyl ester to give the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.88(d,1H),7.49(d,1H),7.37(t,1H),7.29(dd,1H),7.2(dd,1H),7.19(t,1H),7.14(t,1H),4.27(t,2H),4.14(t,2H),3.52(s,2H),3.27(t,2H),2.88(t,2H),2.52(t,4H),2.34(s,3H),2.13(qn,2H),2.04(qn,2H),1.69(t,4H)；¹³C NMR(125MHz,DMSO-d₆)δppm 151.5,151.4,148.6,147.3,145.1,140.1,136.7,130.2,129.0,129.0,127.5,126.5,122.5,122.3,119.2,116.5,115.5,115.4,85.9,83.3,68.6,52.3,46.3,43.3,31.1,23.8,23.8,23.0,20.4,12.9；HRMS-ESI(m/z):[M+H]+C₃₅H₃₅FN₇O₃S₂684.2221, found 684.2209.

Example 432- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylbut-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylbut-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A PTFE-coated magnetic stir bar was assembled in a 24mL oven dried vial and 177mg pyrrolidine (2.500mmol,5.0eq.) dissolved in 2.5mL dry THF was added to it. 133mg of 3-bromobut-1-yne (1.0mmol,2.0eq) are then added dropwise via syringe over 5 minutes and stirred at this temperature for 30 minutes. Into the mixture formed301mg of preparation 3a (0.50mmol,1.0eq.) and 18.15mg of Pd (PPh) were added₃)₂Cl₂(0.025mmol,0.05eq.) and 4.76CuI (0.025mmol,0.05eq.) which were then heated to 60 ℃ and stirred at that temperature for 2 hours. The reaction was completely converted. Celite was added to the reaction mixture and volatiles were removed under reduced pressure. It was then purified by flash chromatography using DCM and MeOH (1.2% NH ₃) As eluent, 220mg (73% yield) of the desired product were obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.41(d,1H),7.28(d,1H),7.18(t,1H),4.51(br.,1H),4.26(m,2H),4.13(t,2H),3.77(s,3H),3.5-2.97(br.,4H),3.25(t,2H),2.88(t,2H),2.32(s,3H),2.11(quint.,2H),2.04(m,2H),1.89(br.,4H),1.51(brd.,3H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,129.4,119.6,115.4,68.3,52.4/50.2,52.0,51.7,46.3,30.7,24.2,23.6,23.0,19.7,19.3,15.7。

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylbut-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure II, starting with 220mg of the product of step A (0.47mmol,1.0eq.) and 100mg1, 3-benzothiazol-2-amine (0.668mmol,2.0eq.) gives 150mg (63% yield) of the desired product.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylbut-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₇FN₇O₃S₂698.2377, found 698.2368.

Example 442- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (dimethylamino) ethoxy]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- (3- {4- [2- (dimethylamino) ethoxy group]Phenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy ]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of preparation 3g of the product of preparation (100mg,0.16mmol,1eq) and preparation 4e (56.6mg,0.31mmol,2eq) in tetrahydrofuran (5mL) were added triphenylphosphine (81.9mg,0.31mmol,2eq) and di-tert-butyl azodicarboxylate (71.9mg,0.31mmol,2eq) and the mixture was heated at 50 ℃ for 8 h. The reaction was partitioned between ethyl acetate and brine, and the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-11% methanol in dichloromethane afforded the desired product as a brown gum (35mg,0.04mmol, 28%).

LC/MS(C₄₀H₅₃N₇O₅SiS₂)804[M+H]⁺；RT 2.72(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dt,J＝7.7,0.9Hz,1H),7.48-7.38(m,2H),7.28-7.20(m,1H),6.92-6.83(m,4H),5.85(s,2H),4.33-4.22(m,4H),4.03-3.89(m,4H),3.76-3.66(m,2H),3.30-3.19(m,2H),2.88(t,J＝6.2Hz,2H),2.54(dt,J＝11.9,5.9Hz,2H),2.37(s,3H),2.16(s,6H),2.14-2.00(m,4H),1.30(t,J＝7.1Hz,3H),0.96-0.86(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (dimethylamino) ethoxy]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of the product of step A (35mg,0.04mmol,1eq) in dichloromethane (2.7mL) cooled to 0 deg.C was added trifluoroacetic acid (0.3mL,3.92mmol,90eq) and the mixture was warmed to room temperature and stirred overnight. The reaction solution was diluted with dichloromethane, Cooled to 0 ℃ and neutralized with aqueous ammonia, then the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow solid (21mg,0.03mmol, 72%).

LC/MS(C₃₄H₃₉N₇O₄S₂)674[M+H]⁺；RT 2.12(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.48(br s,1H),7.88(d,J＝7.9Hz,1H),7.50(d,J＝8.0Hz,1H),7.38(td,J＝8.1,7.7,1.3Hz,1H),7.24-7.15(m,1H),6.93-6.82(m,4H),5.77(s,1H),4.33-4.23(m,4H),4.02-3.90(m,4H),3.31-3.23(m,2H),2.89(t,J＝6.3Hz,2H),2.58-2.51(m,2H),2.35(s,3H),2.17(s,6H),2.13-2.01(m,4H),1.31(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (dimethylamino) ethoxy]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (21mg,0.03mmol,1eq) in 1, 4-dioxane (2mL) was added lithium hydroxide monohydrate (13.1mg,0.31mmol,10eq) and the mixture was heated at reflux for 15 h. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-20% 7N methanolic ammonia in dichloromethane afforded the desired product as a white solid (14.6mg,0.02mmol, 73%).

HRMS-ESI(m/z)[M+H]+C₃₂H₃₆N₇O₄S₂646.2270, found 646.2292.

Example 455- (3- {4- [3- (azetidin-1-yl) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

Step A5- [3- [4- [3- (azetidin-1-yl) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using propynylamine preparation general procedure, starting with 258mg of preparation 3d (0.40mmol,1eq.) as the appropriate propargyl alcohol and azetidine (456.8mg,20eq.) gives 36mg of the desired product (36%).

Step B5- (3- {4- [3- (azetidin-1-yl) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -1, 3-thiazole-4-carboxylic acid

Using the general procedure for hydrolysis, the starting material used the product of step A as the appropriate methyl ester to give the desired product.

HRMS-ESI(m/z):[M+H]+C₃₄H₃₃FN₇O₃S₂670.2064, found 670.2065.

Example 462- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (1-piperidinyl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (1-piperidinyl) prop-1-ynyl ]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for propynylamine, starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and piperidine (264.2mg,20eq.) gives 55mg of the desired product (50%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (1-piperidinyl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

The general procedure for hydrolysis was used with the starting material using the product of step a as the appropriate methyl ester to give the desired product.

HRMS-ESI(m/z):[M+H]+C₃₆H₃₇FN₇O₃S₂698.2377, found 698.2373.

Example 472- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-morpholinoprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-morpholinoprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and morpholine (270.3mg,20eq.) to give 191mg of the desired product (86%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-morpholinoprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₅H₃₅FN₇O₄S₂700.2170, found 700.2163.

Example 482- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and using 1-methylpiperazine (310.7mg,20eq.) to give 150mg of the desired product (79%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) prop-1-ynyl]Phenoxy radical ]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₈FN₈O₃S₂713.2486, found 713.2474.

Example 492- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (4, 4-difluoropiperidin-1-yl) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (4, 4-difluoro-1-piperidinyl) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and 4, 4-difluoropiperidine (20eq.) to give 120mg of the desired product (72%).

Step B2- {3- [ (1, 3-benzo ]Thiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (4, 4-difluoropiperidin-1-yl) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₅F₃N₇O₃S₂734.2189, found 734.2185.

Example 502- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (3, 3-difluoropiperidin-1-yl) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (3, 3-difluoro-1-piperidinyl) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for preparation of propynylamine, starting from 100mg of preparation 3d (0.155mmol,1eq.) as appropriate propargyl alcohol and using 3, 3-difluoropiperidine hydrochloride (1:1) (488.9mg,20eq.) gives 30mg of the desired product (26%).

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (3, 3-difluoropiperidin-1-yl) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₅F₃N₇O₃S₂734.2189, found 734.2186.

Example 51:2- {3- [ (1, 3-benzothiazol-2-yl) aminoBase of]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]Pyridazin-8-yl } -5- (3- { 2-chloro-4- [3- (dimethylamino) prop-1-yn-1-yl-)]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- [3- (2-chloro-4-iodophenoxy) propyl group]-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (225mg,0.32mmol,1eq) in toluene (10mL) was added 2-chloro-4-iodophenol (100mg,0.39mmol,1.22eq), followed by triphenylphosphine (127mg,0.48mmol,1.5eq) and diisopropyl azodicarboxylate (0.1mL,0.48mmol,1.5eq) and the mixture was stirred at room temperature for 3 hours. Purification by flash chromatography (20g silica gel) eluting with a gradient of 0-30% ethyl acetate in isoheptane afforded the desired product as a yellow solid (235mg,0.23mmol, 72%).

LC/MS(C₃₅H₄₀ClIN₆O₄SiS₂)863[M+H]⁺；RT 1.48(LCMS-V-B2)

¹H NMR(400MHz,CDCl₃)δ7.65(d,J＝2.1Hz,1H),7.59(dt,J＝7.7,0.9Hz,1H),7.47(dd,J＝8.6,2.2Hz,1H),7.39-7.35(m,2H),7.24-7.17(m,1H),6.67(d,J＝8.6Hz,1H),5.84(s,2H),4.47-4.35(m,2H),4.13-4.06(m,2H),3.90(s,3H),3.80-3.70(m,2H),3.43-3.34(m,2H),2.87(t,J＝6.3Hz,2H),2.38(s,3H),2.34-2.22(m,2H),2.19-2.06(m,2H),1.02-0.91(m,2H),-0.07(s,9H)。

Step B5- (3- { 2-chloro-4- [3- (dimethylamino) prop-1-yn-1-yl)]Phenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

Dimethyl (prop-2-yn-1-yl) amine (0.1mL,0.93mmol,4.0eq) was added to a solution of the product of step a (235mg,0.23mmol,1eq), copper (I) iodide (8.81mg,0.05mmol,0.2eq) and tetrakis (triphenylphosphine) palladium (0) (26.7mg,0.02mmol,0.1eq) in 2-methyltetrahydrofuran (10mL), followed by N, N-diisopropylethylamine (0.15mL,0.69mmol,3eq) and the mixture was heated at 75 ℃ for 18 h. The reaction was cooled to room temperature and purified by flash column chromatography (20g silica gel) eluting with a gradient of 0-100% ethyl acetate in isoheptane to give the desired product as a brown gum (75mg,0.09mmol, 40%).

LC/MS(C₄₀H₄₈ClN₇O₄SiS₂)818[M+H]⁺；RT 1.56(LCMS-V-B1)

¹H NMR(400MHz,CDCl₃)δ7.70-7.63(m,1H),7.59(dt,J＝7.6,0.9Hz,1H),7.48-7.44(m,1H),7.40-7.35(m,2H),7.23-7.16(m,1H),6.84(d,J＝8.6Hz,1H),5.84(s,2H),4.46-4.36(m,2H),4.18-4.05(m,2H),3.90(s,3H),3.79-3.69(m,2H),3.49-3.34(m,4H),2.87(t,J＝6.5Hz,2H),2.39(s,3H),2.35(s,6H),2.32-2.25(m,2H),2.18-2.08(m,2H),1.01-0.93(m,2H),-0.07(s,9H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-chloro-4- [3- (dimethylamino) prop-1-yn-1-yl-)]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (75mg,0.09mmol,1eq) cooled to 0 ℃ in dichloromethane (1.8mL) was added trifluoroacetic acid (0.2mL,2.75mmol,30eq) and the mixture was stirred for 36 h. The reaction was diluted with dichloromethane (20mL), washed sequentially with 1M aqueous ammonia (10mL), water (10mL) and brine (10mL), dried (magnesium sulfate) and concentrated in vacuo. The crude product was suspended in a mixture of water (1.5mL) and methanol (0.5mL), lithium hydroxide monohydrate (11.5mg,0.27mmol,3eq) was added and the suspension was heated at 80 ℃ for 18 h. The reaction was neutralized with acetic acid and the solid was collected by filtration and washed with water (20 mL). Purification by preparative HPLC (HPLC-V-A2) gave the desired product as a pale yellow solid (7mg,0.01mmol, 11%).

HRMS-ESI(m/z)[M+H]+C₃₃H₃₃ClN₇O₃S₂674.1775, found 674.1796.

Examples52:2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (pyrrolidin-1-yl) ethoxy]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy ]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- (3- {4- [2- (pyrrolidin-1-yl) ethoxy]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To a solution of preparation 3g of the product of preparation (150mg,0.23mmol,1eq) and preparation 4f of the product of preparation (97mg,0.47mmol,2eq) in toluene (6mL) were added triphenylphosphine (123mg,0.47mmol,2eq) and di-tert-butyl azodicarboxylate (108mg,0.47mmol,2eq) and the mixture was heated at 50 ℃ overnight. The reaction was partitioned between dichloromethane and brine, the organic phase was dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-12% methanol in dichloromethane afforded the desired product as a yellow gum (165mg,0.2mmol, 85%).

LC/MS(C₄₂H₅₅N₇O₅SiS₂)831[M+H]⁺；RT 2.75(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(d,1H),7.48-7.39(m,2H),7.28-7.20(m,1H),6.92-6.82(m,4H),5.84(s,2H),4.34-4.21(m,4H),4.02-3.91(m,4H),3.76-3.66(m,2H),3.26(t,2H),2.87(t,J＝6.2Hz,2H),2.71(t,1H),2.48-2.42(m,4H),2.37(s,3H),2.13-2.00(m,4H),1.71-1.60(m,4H),1.30(t,J＝7.1Hz,3H),0.95-0.86(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (pyrrolidin-1-yl) ethoxy]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid ethyl ester

To the product of step A (165mg,0.2mmol,1eq) cooled to 0 ℃ was added dichloroTo a solution of methane (8.1mL) was added trifluoroacetic acid (0.91mL,11.9mmol,60eq) and the mixture was stirred at room temperature overnight. The reaction was diluted with dichloromethane, cooled to 0 ℃, neutralized with aqueous ammonia, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMCartridge silica) and gradient eluted with 0-14% methanol in dichloromethane afforded the desired product as a yellow solid (84mg,0.12mmol, 60%).

LC/MS(C₃₆H₄₁N₇O₄S₂)700[M+H]⁺；RT 2.14(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.48(br s,1H),7.89(d,J＝7.7Hz,1H),7.50(d,J＝7.9Hz,1H),7.42-7.34(m,1H),7.20(td,J＝7.5,1.2Hz,1H),6.94-6.83(m,4H),4.34-4.21(m,4H),3.98(t,J＝6.2Hz,4H),3.32-3.23(m,2H),2.89(t,J＝6.4Hz,2H),2.85-2.74(m,2H),2.65-2.53(m,4H),2.35(s,3H),2.16-1.99(m,4H),1.70(s,4H),1.31(t,J＝7.1Hz,3H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (pyrrolidin-1-yl) ethoxy]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (84mg,0.12mmol,1eq) in 1, 4-dioxane (4mL) was added lithium hydroxide monohydrate (50.4mg,1.2mmol,10eq) and the mixture was heated at reflux overnight. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-15% 7N methanolic ammonia in dichloromethane afforded a solid which was triturated with ether, filtered and dried in vacuo to afford the desired product as a yellow solid (52.9mg,0.08mmol, 66%).

LC/MS(C₃₄H₃₇N₇O₄S₂)672[M+H]⁺；RT 1.93(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.89(dd,J＝7.9,1.2Hz,1H),7.50(d,J＝8.1Hz,1H),7.38(td,J＝7.7,1.3Hz,1H),7.20(td,J＝7.6,1.2Hz,1H),6.94-6.81(m,4H),4.28(dd,J＝7.3,4.3Hz,2H),4.05-3.91(m,4H),3.35-3.17(m,6H),2.89(t,J＝6.2Hz,2H),2.76(t,J＝5.9Hz,2H),2.35(s,3H),2.18-1.96(m,4H),1.73-1.62(m,4H)。

HRMS-ESI(m/z)[M+H]+C₃₄H₃₈N₇O₄S₂Calculated value of (2) 672.2427, found value of 672.2449

Example 532- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (dimethylamino) ethyl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- (3- {4- [2- (dimethylamino) ethyl]-2-fluorophenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy ]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (95mg,0.15mmol,1eq) and preparation 4g (41mg,0.22mmol,1.48eq) in toluene (5mL) was added di-tert-butyl azodicarboxylate (69.8mg,0.3mmol,2eq) and triphenylphosphine (79.5mg,0.3mmol,2eq) and the mixture was heated at 50 ℃ for 20 h. The reaction was partitioned between dichloromethane and water, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-5% methanol in dichloromethane afforded the desired product as a yellow oil (83mg,0.1mmol, 69%).

LC/MS(C₃₉H₅₀FN₇O₄SiS₂)792[M+H]⁺；RT 2.69(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dd,J＝7.3,1.1Hz,1H),7.48-7.38(m,2H),7.28-7.20(m,1H),7.14-7.02(m,2H),6.98-6.92(m,1H),5.85(s,2H),4.26(dd,J＝7.2,4.4Hz,2H),4.08(t,J＝6.2Hz,2H),3.79(s,3H),3.75-3.67(m,2H),3.31-3.23(m,2H),2.88(t,J＝6.4Hz,2H),2.65-2.56(m,2H),2.46-2.37(m,2H),2.37(s,3H),2.12(s,6H),2.11-2.07(m,2H),2.07-2.00(m,2H),0.96-0.88(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (dimethylamino) ethyl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step A (83mg,0.1mmol,1eq) in dichloromethane (5mL) was added trifluoroacetic acid (1mL,12.99mmol,124eq) and the mixture was stirred at room temperature overnight. The reaction was diluted with dichloromethane, cooled to 0 ℃, neutralized with aqueous ammonia, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep) ^TMCartridge silica) and gradient eluted with 0-8% methanol in dichloromethane afforded the desired product as a yellow solid (50mg,0.08mmol, 72%).

LC/MS(C₃₃H₃₆FN₇O₃S₂)662[M+H]⁺；RT 2.04(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.48(br s,1H),7.88(d,J＝7.9Hz,1H),7.49(d,J＝8.0Hz,1H),7.40-7.34(m,1H),7.19(td,J＝7.6,1.2Hz,1H),7.13-7.02(m,2H),6.98-6.92(m,1H),4.30-4.21(m,2H),4.08(t,2H),3.78(s,3H),3.32-3.24(m,2H),2.87(t,J＝6.3Hz,2H),2.62(dd,J＝8.6,6.6Hz,2H),2.46-2.38(m,2H),2.34(s,3H),2.14(s,6H),2.13-2.09(m,2H),2.08-2.01(m,2H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [2- (dimethylamino) ethyl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (50mg,0.08mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (31.7mg,0.76mmol,10eq) and the mixture was heated at 70 ℃ overnight. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-15% 7N methanolic ammonia in dichloromethane afforded the desired product as a yellow solid (24.68mg,0.04mmol, 50%).

LC/MS(C₃₂H₃₄FN₇O₃S₂)648[M+H]⁺；RT 1.87(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.89(d,J＝7.8Hz,1H),7.50(d,J＝8.1Hz,1H),7.38(td,J＝8.1,7.7,1.3Hz,1H),7.20(td,J＝7.5,1.2Hz,1H),7.14-7.02(m,2H),6.95(dt,J＝8.4,1.4Hz,1H),4.32-4.25(m,2H),4.09(t,J＝6.3Hz,2H),3.31-3.23(m,2H),2.89(t,J＝6.3Hz,2H),2.63(t,J＝7.6Hz,2H),2.44(dd,J＝8.5,6.6Hz,2H),2.35(s,3H),2.17(s,6H),2.15-2.08(m,2H),2.08-2.00(m,2H)

HRMS-ESI(m/z)[M+H]+C₃₂H₃₅FN₇O₃S₂648.2227, found 648.2269.

Example 543- [2- [3- [4- [3- [2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-4-carboxy-thiazol-5-yl]Propoxy group]-3-fluoro-phenyl]Prop-2-ynylamino]Ethyl-dimethyl-ammonium radical]Propane-1-sulfonic acid salt

Step A3- [2- [ tert-butoxycarbonyl- [3- [4- [3- [2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] ]Pyridazin-8-yl) -4-methoxycarbonyl-thiazol-5-yl]Propoxy group]-3-fluoro-phenyl]Prop-2-ynyl]Amino group]Ethyl-dimethyl-ammonium radical]Propane-1-sulfonic acid salt

A mixture of 384mg of example 76 (step A) (0.55mmol,1eq.) and 1944mg of oxathiolane 2, 2-dioxide (15.92mmol,30eq.) in acetonitrile (4mL/mmol) and DMF (1mL/mmol) is stirred at room temperature. After reaching the appropriate conversion, the volatiles were removed under reduced pressure and purified by flash column chromatography (SiO)₂,EtOAc:0.6M NH₃MeOH solution) to yield 94mg (29%) of the desired product.

LC-MS-ESI(m/z):[M+H]⁺C₃₇H₄₉ClFN₆O₈S₂823.3, found 823.2.

Step B3- [2- [3- [4- [3- [2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-4-methoxycarbonyl-thiazol-5-yl]Propoxy group]-3-fluoro-phenyl]Prop-2-ynyl-tert-butoxycarbonyl-amino]Ethyl-dimethyl-ammonium radical]Propane-1-sulfonic acid salt

Using Buchwald general procedure II, starting from the product of step A and 1, 3-benzothiazol-2-amine, the desired product was obtained.¹H NMR(500MHz,DMSO-d₆)δppm 7.88(d,1H),7.49(d,1H),7.37(t,1H),7.35(d,1H),7.25(dd,1H),7.20(t,1H),7.18(t,1H),4.27(s,2H),4.26(t,2H),4.15(t,2H),3.77(s,3H),3.69(t,2H),3.46(t,2H),3.45(t,2H),3.28(t,2H),3.07(s,6H),2.88(t,2H),2.46(t,2H),2.34(s,3H),2.13(qn,2H),2.04(qn,2H),2.02(qn,2H),1.44(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 129.2,126.5,122.6,122.4,119.3,116.9,115.5,85.4,82.5,68.4,63.3,60.1,52.0,50.8,48.0,46.4,40.8,37.9,31.0,28.4,23.9,23.2,20.4,19.5,13.0；HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₄FN₈O₈S₃937.3205, found 937.3209.

Step C3- [2- [3- [4- [3- [2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-4-carboxy-thiazol-5-yl]Propoxy group]-3-fluoro-phenyl ]Prop-2-ynylamino]Ethyl-dimethyl-ammonium radical]Propane-1-sulfonic acid salt

Using the general procedure of deprotection and hydrolysis, followed by purification again by reverse phase preparative chromatography (C18,25mM NH)₄HCO₃MeCN) using the product of step B as starting material to obtain the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₈H₄₄FN₈O₆S₃823.2524, found 823.2523.

Example 552- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazine-8-

Yl ] -5- [3- [ 2-fluoro-4- [3- (3-hydroxy-1-piperidinyl) prop-1-ynyl ] phenoxy ] propyl ] thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-hydroxy-1-piperidinyl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting from 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and using piperidin-3-ol (313.8mg,20eq.) to give 70mg of the desired product (62%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-hydroxy-1-piperidinyl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₇FN₇O₄S₂714.2327, found 714.2323.

Example 565- [3- [4- [3- [ (1S,5R) -3-azabicyclo [3.1.0]]Hex-3-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

Step A5- [3- [4- [3- [ (1R,5S) -3-azabicyclo [3.1.0]]Hex-3-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for preparation of propynylamine, starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and using (1R,5S) -3-azabicyclo [3.1.0] hexane (20eq.) gives 150mg of the desired product (81%).

Step B5- [3- [4- [3- [ (1S,5R) -3-azabicyclo [3.1.0]]Hex-3-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₅FN₇O₃S₂696.2221, found 696.2227.

Example 572- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] ]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- [4- (1-piperidinyl) -1-piperidinyl]Prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- [4- (1-piperidinyl) -1-piperidinyl]Prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for preparation of propynylamine, starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and using 1- (4-piperidinyl) piperidine hydrochloride (1:2) (748.3mg,20eq.) gives 100mg of the desired product (81%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- [4- (1-piperidinyl) -1-piperidinyl]Prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₄₁H₄₆FN₈O₃S₂781.3112, found 781.3112.

Example 582- [3- (1, 3-benzothiazole-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-oxo-2, 8-diazaspiro [4.5 ]]Dec-8-yl) prop-1-ynyl]Phenoxy radical]Propyl radical ]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-oxo-2, 8-diazaspiro [4.5]]Dec-8-yl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and 2, 8-diazaspiro [4.5] decan-3-one (478.4mg,20eq.) to give 125mg of the desired product (82%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-oxo-2, 8-diazaspiro [4.5]]Dec-8-yl) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₉H₄₀FN₈O₄S₂767.2592, found 767.2588.

Example 592- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-bromo-4- [3- (dimethylamino) prop-1-yn-1-yl-)]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- [3- (2-bromo-4-iodophenoxy) propyl]-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy group ]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino group } -5H, and a pharmaceutically acceptable salt thereof,6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (350mg,0.5mmol,1eq) and 2-bromo-4-iodophenol (200mg,0.67mmol,1.33eq) in toluene (10mL) was added triphenylphosphine (198mg,0.75mmol,1.5eq) and diisopropyl azodicarboxylate (0.15mL,0.75mmol,1.5eq) and the mixture was stirred at room temperature for 18 h. The reaction was concentrated in vacuo and purified by flash column chromatography (20g silica gel) with a gradient elution of 0-30% ethyl acetate in isoheptane to afford the desired product as a yellow solid (555mg,0.49mmol, 97%).

LC/MS(C₃₅H₄₀BrIN₆O₄SiS₂)907[M+H]⁺；RT 1.50(LCMS-V-B2)

¹H NMR(400MHz,CDCl₃)δ7.82(d,J＝2.1Hz,1H),7.59(dt,J＝7.6,0.9Hz,1H),7.50(dd,J＝8.6,2.1Hz,1H),7.39-7.34(m,2H),7.24-7.16(m,1H),6.65(d,J＝8.7Hz,1H),5.84(s,2H),4.47-4.36(m,2H),4.17-4.05(m,2H),3.90(s,3H),3.80-3.69(m,2H),3.39(dd,J＝8.4,6.6Hz,2H),2.87(t,J＝6.4Hz,2H),2.38(s,3H),2.34-2.23(m,2H),2.18-2.07(m,2H),1.00-0.92(m,2H),-0.07(s,9H)。

Step B5- (3- { 2-bromo-4- [3- (dimethylamino) prop-1-yn-1-yl)]Phenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step A (550mg,0.52mmol,1eq), copper (I) iodide (18.5mg,0.1mmol,0.2eq) and tetrakis (triphenylphosphine) palladium (0) (56mg,0.05mmol,0.1eq) in 2-methyltetrahydrofuran (10mL) was added dimethyl (prop-2-yn-1-yl) amine (0.3mL,2.79mmol,5.75eq) followed by N, N-diisopropylethylamine (0.3mL,1.45mmol,3eq) and the mixture was heated at 75 ℃ for 3 hours. The reaction was cooled to room temperature and concentrated in vacuo. Purification by flash chromatography (20g of silica gel) eluting with a gradient of 0-100% ethyl acetate in isoheptane afforded the desired product as a dark orange gum (220mg,0.25mmol, 53%).

LC/MS(C₄₀H₄₈BrN₇O₄SiS₂)862[M+H]⁺；RT 1.54(LCMS-V-B2)

¹H NMR(400MHz,CDCl₃)δ7.64(d,J＝2.0Hz,1H),7.60(dt,J＝7.7,0.9Hz,1H),7.38-7.37(m,2H),7.34-7.30(m,1H),7.23-7.16(m,1H),6.81(d,J＝8.6Hz,1H),5.84(s,2H),4.48-4.35(m,2H),4.16-4.07(m,2H),3.90(s,3H),3.78-3.69(m,2H),3.50-3.35(m,4H),2.87(t,J＝6.3Hz,2H),2.38(s,3H),2.35(s,6H),2.33-2.26(m,2H),2.18-2.07(m,2H),1.01-0.88(m,2H),-0.07(s,9H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-bromo-4- [3- (dimethylamino) prop-1-yn-1-yl-)]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (210mg,0.24mmol,1eq) in dichloromethane (4.5mL) cooled to 0 deg.C was added trifluoroacetic acid (0.5mL,6.08mmol,25eq) and the mixture was stirred at room temperature for 24 h. The reaction was diluted with dichloromethane (40mL), washed sequentially with 1M aqueous ammonia (20mL), water (2 × 20mL) and brine (20mL), dried (magnesium sulfate) and concentrated in vacuo. To a solution of the crude product in methanol (1mL) was added water (2mL) and lithium hydroxide monohydrate (30.6mg,0.73mmol,3eq) and the suspension was heated at 75 ℃ for 72 h. The reaction was cooled to room temperature and then neutralized with acetic acid, the solid was collected by filtration and washed with water (2 × 10 mL). Purification by preparative HPLC (HPLC-V-a2) gave the desired product as a green solid (15mg,0.02mmol, 9%).

LC/MS(C₃₃H₃₂BrN₇O₃S₂)718[M+H]⁺；RT 1.18(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.87(d,J＝7.8Hz,1H),7.63(d,J＝2.0Hz,1H),7.49(d,J＝8.0Hz,1H),7.42-7.33(m,2H),7.20(td,J＝7.6,1.1Hz,1H),7.09(d,J＝8.6Hz,1H),4.26(t,J＝5.5Hz,2H),4.13(t,J＝6.1Hz,2H),3.34-3.25(m,2H),2.86(t,J＝6.1Hz,2H),2.54(s,2H),2.33(s,3H),2.17(s,6H),2.16-2.09(m,2H),2.08-1.95(m,2H)。

Example 602- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ 2-fluoro-4- (4-methylpiperazin-1-yl) phenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Step A5- [3- (2-fluoro-4-iodophenoxy) propyl ]-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (200mg,0.32mmol,1eq) and 2-fluoro-4-iodophenol (152mg,0.64mmol,2eq) in toluene (6mL) was added triphenylphosphine (167mg,0.64mmol,2eq) and diisopropyl azodicarboxylate (147mg,0.64mmol,2eq) and the mixture was stirred at 50 ℃ for 17 h. The reaction was partitioned between dichloromethane and water, the organic phase was separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a yellow gum (161mg,0.19mmol, 60%).

LC/MS(C₃₅H₄₀FIN₆O₄SiS₂)847[M+H]⁺；RT 3.33(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dt,J＝7.6,0.9Hz,1H),7.61(dd,J＝10.8,2.1Hz,1H),7.49-7.39(m,3H),7.24(ddd,J＝8.3,6.6,1.9Hz,1H),7.01(t,J＝8.8Hz,1H),5.85(s,2H),4.26(t,J＝5.7Hz,2H),4.11(t,J＝6.2Hz,2H),3.78(s,3H),3.76-3.67(m,2H),3.27(t,2H),2.88(t,J＝6.3Hz,2H),2.37(s,3H),2.18-2.09(m,2H),2.08-2.01(m,2H),0.95-0.86(m,2H),-0.12(s,9H)。

Step B5- {3- [ 2-fluoro-4- (4-methylpiperazin-1-yl) phenoxy group]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

1-Methylpiperazine (14.5. mu.L, 0.13mmol,1.5eq) was added to the product of step A (74mg,0.09mmol,1eq), copper (I) iodide (1.66mg,0.01mmol,0 1eq), tripotassium phosphate (37.1mg,0.17mmol,2eq) and [ (2, 6-dimethylphenyl) carbamoyl]Formic acid (3.38mg,0.02mmol,0.2eq) in DMSO (2mL) and the mixture was heated under microwave irradiation at 120 ℃ for 2 hours. The reaction was cooled to room temperature, partitioned between ethyl acetate and water, the organic phase was washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMCartridge silica) and gradient eluted with 0-8% methanol in dichloromethane afforded the desired product as a brown gum (30mg,0.04mmol, 42%).

LC/MS(C₄₀H₅₁FN₈O₄SiS₂)819[M+H]⁺；RT 2.71(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(d,1H),7.48-7.38(m,2H),7.27-7.20(m,1H),7.03(t,1H),6.84(dd,1H),6.67-6.61(m,1H),5.84(s,2H),4.26(t,J＝5.7Hz,2H),4.02(t,J＝6.2Hz,2H),3.79(s,3H),3.76-3.66(m,2H),3.26(t,2H),3.04-2.95(m,4H),2.92-2.82(m,2H),2.42-2.32(m,7H),2.18(s,3H),2.12-1.99(m,4H),0.95-0.86(m,2H),-0.11(s,9H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ 2-fluoro-4- (4-methylpiperazin-1-yl) phenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step B (116mg,0.14mmol,1eq) cooled to 0 ℃ in dichloromethane (7.5mL) was added trifluoroacetic acid (1.52mL,19.8mmol,140eq) and the mixture was stirred at room temperature for 24 h. The reaction was diluted with dichloromethane (40mL), washed with aqueous ammonia, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow gum (72mg,0.1mmol, 74%).

LC/MS(C₃₄H₃₇FN₈O₃S₂)689[M+H]⁺；RT 2.01(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ11.46(br s,1H),7.89(d,J＝7.8Hz,1H),7.56-7.44(m,1H),7.38(ddd,J＝8.3,7.3,1.3Hz,1H),7.24-7.16(m,1H),7.04(dd,J＝9.9,9.0Hz,1H),6.85(dd,J＝14.6,2.8Hz,1H),6.69-6.61(m,1H),4.27(t,J＝5.8Hz,2H),4.08-3.99(m,2H),3.80(s,3H),3.31-3.23(m,2H),3.05-2.96(m,4H),2.89(t,J＝6.3Hz,2H),2.42-2.36(m,4H),2.35(s,3H),2.19(s,3H),2.13-2.00(m,4H)。

Step D2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ 2-fluoro-4- (4-methylpiperazin-1-yl) phenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step C (72mg,0.1mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (43.9mg,1.05mmol,10eq) and the mixture was heated at reflux for 3 h. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-15% 7N methanolic ammonia in dichloromethane to give a solid which was triturated with ether, filtered and dried in vacuo to give the desired product as a yellow solid (35mg,0.05mmol, 50%).

HRMS-ESI(m/z)[M+H]+C₃₃H₃₆FN₈O₃S₂675.2336, found 675.2364

Example 612- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- [3- ({3- [ (dimethylamino) methyl group]-5-fluoro-1-methyl-1H-indol-6-yl } oxy) propyl]-1, 3-thiazole-4-carboxylic acid

Step A5- [3- ({3- [ (dimethylamino) methyl group)]-5-fluoro-1-methyl-1H-indol-6-yl } oxy) propyl]-2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c) ]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

Dissolve the product of preparation 3b (120mg,0.19mmol,1eq) in toluene (15mL) and add the product of preparation 4h (67mg,0.26mmol,1.35eq), then triphenylphosphine (100mg,0.38mmol,2eq) and occasionallyDi-tert-butyl azodicarboxylate (88.2mg,0.38mmol,2eq) and the mixture was stirred at 50 ℃ overnight. The reaction was partitioned between dichloromethane and water, the organic phase was washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a beige solid (18mg,0.02mmol, 11%).

LC/MS(C₄₁H₅₁FN₈O₄SiS₂)831[M+H]⁺；RT 2.73(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.81(d,J＝7.7Hz,1H),7.47-7.43(m,2H),7.40-7.33(m,1H),7.28-7.21(m,1H),7.15(d,J＝11.6Hz,1H),6.84(d,J＝7.4Hz,1H),5.85(s,2H),4.28(t,2H),4.16(t,J＝6.3Hz,2H),3.79(s,3H),3.76-3.71(m,2H),3.69(s,2H),3.63(s,3H),3.48-3.41(m,4H),3.88(t,2H),2.38(s,3H),2.08-2.00(m,2H),1.46(s,6H),1.35 0.95-0.86(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- [3- ({3- [ (dimethylamino) methyl group]-5-fluoro-1-methyl-1H-indol-6-yl } oxy) propyl]-1, 3-thiazole-4-carboxylic acid

A solution of the product of step A (18mg,0.02mmol,1eq) in dichloromethane (1mL) was cooled to 0 deg.C, trifluoroacetic acid (1mL,13mmol,600eq) was added and the mixture was stirred at room temperature for 4 hours. Dichloromethane (10mL) was added and the solution was cooled to 0 ℃, washed with aqueous ammonia, dried (PTFE phase separator) and concentrated in vacuo. The residue was suspended in 1, 4-dioxane (2mL), lithium hydroxide monohydrate (9.1mg,0.22mmol,10eq) was added and the mixture was heated at 70 ℃ overnight. Purification by preparative HPLC (HPLC-V-B1) gave the desired product (4.5mg,0.01mmol, 30%) as a yellow solid as the formate salt.

HRMS-ESI(m/z)[M+H]+C₃₄H₃₆FN₈O₃S₂687.2336, found 687.2362.

Example 622- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazine-8-5- (3- {4- [4- (dimethylamino) butyl } -phenyl) -methyl- ] -methyl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- (3- {4- [4- (dimethylamino) butyl]-2-fluorophenoxy } propyl) -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (120mg,0.19mmol,1eq) in toluene (5mL) were added the product of preparation 4i (107mg,0.51mmol,2.65eq), di-tert-butyl azodicarboxylate (88mg,0.38mmol,2eq) and triphenylphosphine (100mg,0.38mmol,2eq) and the mixture was heated at 50 ℃ for 20 h. The reaction was partitioned between dichloromethane and water, the organic phase was dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-8% methanol in dichloromethane afforded the desired product as a yellow oil (133mg,0.16mmol, 85%).

LC/MS(C₁₄H₅₄FN₇O₄SiS₂)821[M+H]⁺；RT 2.74(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.85-7.78(m,1H),7.48-7.38(m,2H),7.27-7.20(m,1H),7.11-7.01(m,2H),6.95-6.89(m,1H),5.84(s,2H),4.26(t,J＝6.0Hz,2H),4.07(t,2H),3.78(s,3H),3.71(dd,2H),3.28(dd,J＝15.9,8.4Hz,2H),2.87(t,J＝6.2Hz,2H),2.37(s,3H),2.21-2.08(m,4H),2.08-2.01(m,8H),1.55-1.42(m,2H),1.41-1.27(m,2H),0.95-0.87(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino ]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [4- (dimethylamino) butyl-)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

A solution of the product of step A (133mg,0.16mmol,1eq) in dichloromethane (6mL) was cooled to 0 deg.C and trifluoroacetic acid (1.2) was added4mL,16.2mmol,100eq) and the mixture was stirred at room temperature for 24 hours. Dichloromethane (40mL) was added and the solution was washed with saturated aqueous ammonium chloride, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-14% methanol in dichloromethane afforded the desired product as a yellow gum (81mg,0.12mmol, 72%).

LC/MS(C₃₅H₄₀FN₇O₃S₂)690[M+H]⁺；RT 2.14(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.87(dd,J＝7.8,1.2Hz,1H),7.49(d,J＝8.0Hz,1H),7.37(ddd,J＝8.2,7.2,1.3Hz,1H),7.19(td,J＝7.5,1.2Hz,1H),7.09-7.01(m,2H),6.94-6.87(m,1H),4.25(dd,J＝7.3,4.5Hz,2H),4.07(t,J＝6.2Hz,2H),3.78(s,3H),3.27(t,J＝7.7Hz,2H),2.87(t,J＝6.3Hz,2H),2.48-2.43(m,2H),2.33(s,3H),2.18-2.07(m,4H),2.08-1.98(m,8H),1.55-1.41(m,2H),1.40-1.26(m,2H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [4- (dimethylamino) butyl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (81mg,0.12mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (49.3mg,1.17mmol,10eq) and the mixture was heated at 70 ℃ overnight. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-20% 7N methanolic ammonia in dichloromethane afforded the desired product as a yellow solid (48.3mg,0.07mmol, 61%).

HRMS-ESI(m/z)[M+H]+C₃₄H₃₉FN₇O₃S₂676.2540, found 676.2569.

Example 632- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for preparation of propynylamine, starting with 100mg of preparation 3d (0.155mmol,1eq.) as the appropriate propargyl alcohol and using (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0] hexane (20eq.) gives 61mg of the desired product (52%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₃F₃N₇O₃S₂732.2033, found 732.2023.

Example 642- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [2- (methylamino) ethyl group]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- {3- [4- (2- { [ (tert-butoxy) carbonyl](methyl) amino } ethyl) -2-fluorophenoxy]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (80mg,0.13mmol,1eq) in toluene (5mL) was addedPrepared product of example 4j (48mg,0.18mmol,1.4eq), di-tert-butyl azodicarboxylate (58.8mg,0.26mmol,2eq) and triphenylphosphine (67mg,0.26mmol,2eq) and the mixture was heated at 50 ℃ overnight. The reaction was partitioned between dichloromethane and water, separated (phase separator) and the organic phase concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a colourless gum (77mg,0.09mmol, 69%).

LC/MS(C₄₃H₅₆FN₇O₆SiS₂)878[M+H]⁺；RT 3.28(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dt,J＝7.6,0.9Hz,1H),7.48-7.38(m,2H),7.24(ddd,J＝8.3,6.7,1.8Hz,1H),7.13-7.01(m,2H),6.96-6.86(m,1H),5.84(s,2H),4.27(t,J＝5.7Hz,2H),4.08(t,2H),3.78(s,2H),3.75-3.67(m,2H),3.31-3.22(m,4H),2.88(t,J＝6.2Hz,2H),2.71(s,3H),2.69-2.61(m,2H),2.37(s,3H),2.17-2.06(m,2H),2.05-2.00(m,2H),1.24(s,9H),0.95-0.86(m,2H),-0.12(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [2- (methylamino) ethyl ] ethyl]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

A solution of the product of step A (77mg,0.09mmol,1eq) in dichloromethane (4mL) was cooled to 0 deg.C, trifluoroacetic acid (0.81mL,10.5mmol,120eq) was added and the mixture was stirred at room temperature for 24 h. The reaction was diluted with dichloromethane, washed with aqueous ammonia, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-18% methanol in dichloromethane afforded the desired product as a yellow solid (39mg,0.06mmol, 69%).

LC/MS(C₃₂H₃₄FN₇O₃S₂)648[M+H]⁺；RT 2.02(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.91-7.84(m,1H),7.49(d,J＝8.0Hz,1H),7.41-7.33(m,1H),7.19(td,J＝7.5,1.2Hz,1H),7.12-7.03(m,2H),6.94(dd,J＝8.8,1.9Hz,1H),4.27(dd,J＝7.1,4.5Hz,2H),4.09(t,J＝6.2Hz,2H),3.79(s,3H),3.31-3.21(m,2H),2.88(t,J＝6.2Hz,2H),2.68-2.57(m,4H),2.35(s,3H),2.25(s,3H),2.17-2.01(m,4H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [2- (methylamino) ethyl group]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (39mg,0.06mmol,1eq) in 1, 4-dioxane (2mL) was added lithium hydroxide monohydrate (25.3mg,0.6mmol,10eq) and the mixture was heated at reflux for 4 h. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) was purified, and the solid was obtained by gradient elution with 0-25% 7N methanolic ammonia in dichloromethane, suspended in ethyl acetate (2mL) and added with hydrochloric acid (4M in 1, 4-dioxane; 68.9. mu.L, 0.28mmol,4.58 eq). The mixture was stirred for 10 min, then the solid was collected by filtration and dried in vacuo to give the desired product as a yellow solid (17.2mg,0.03mmol, 45%) as the hydrochloride salt.

HRMS-ESI(m/z)[M+H]+C₃₁H₃₃FN₇O₃S₂634.2070, found 634.2093.

Example 65:2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6- [2- (methylamino) ethoxy]-5H,6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A4-methylmorpholin-3-one

A solution of 2- (methylamino) ethanol (5.32mL,66.6mmol,1eq) in ethanol (100mL) and 35% aqueous sodium hydroxide (6.25mL) was cooled to 15-20 deg.C and chloroacetyl chloride (13.3mL,166mmol,2.5eq) and 35% aqueous sodium hydroxide (22mL) were added simultaneously over 1 hour with vigorous stirring. The mixture was stirred for 20 minutes and then acidified with hydrochloric acidThe solution was neutralized and extracted with dichloromethane (3x100 mL). The combined organic extracts were washed with water, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a colorless oil (4.4g,38.2mmol, 58%).

¹H NMR(400MHz,DMSO-d6)δ4.00(s,2H),3.84-3.78(m,2H),3.36-3.29(m,2H),2.86(s,3H)。

Step B2- (but-2-yn-1-yl) -4-methylmorpholin-3-one

To a solution of diisopropylamine (6.45mL,45.9mmol,1.2eq) in tetrahydrofuran (130mL) cooled to-78 deg.C was added n-butyllithium (2.06M in hexane; 20.4mL,42mmol,1.1eq) dropwise. After 1 min a solution of the product of step A (4.4g,38.2mmol,1eq) in tetrahydrofuran (30mL) was added dropwise. After 15 min, a solution of 1-bromo-2-butyne (4.02mL,45.9mmol,1.2eq) in tetrahydrofuran (15mL) was added dropwise and the mixture was stirred at-78 deg.C for 1 h, then warmed to room temperature. Saturated aqueous ammonium chloride was added and the mixture was extracted with ethyl acetate (× 3), the combined organic extracts were dried (magnesium sulphate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80g RediSep) ^TMCartridge silica) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a yellow oil (5.15g,30.8mmol, 81%).

¹H NMR(400MHz,DMSO-d6)δ4.09(dd,J＝7.6,3.5Hz,1H),4.01-3.94(m,1H),3.76(ddd,J＝11.9,10.0,3.6Hz,1H),3.52-3.41(m,1H),3.26-3.18(m,1H),2.86(s,3H),2.67-2.58(m,1H),2.57-2.44(m,1H),1.73(t,J＝2.6Hz,3H)。

Step C2- [2- (methylamino) ethoxy group]Hex-4-ynoic acid

To a solution of the product of step B (3.25g,19.4mmol,1eq) in methanol (110mL) was added 1M aqueous lithium hydroxide (60.3mL,60.3mmol,3.1eq) and the mixture was heated at reflux overnight. The reaction was concentrated in vacuo to afford the desired product as an orange gum (5.15g,27.8mmol, 100%) which was used directly in the next step without further characterization.

Step D:2-[2- ({ [ (9H-fluoren-9-yl) methoxy]Carbonyl } (methyl) amino) ethoxy]Hex-4-ynoic acid

To a solution of the product of step C (5.15g,27.8mmol,1eq) in 1, 4-dioxane (45mL) and water (160mL) at 0 deg.C was added potassium carbonate (15.4g,111mmol,4eq) followed by 9H-fluoren-9-yl-methylchloroformate (7.19g,27.8mmol,1eq), and the mixture was warmed to room temperature and stirred for 2 hours. The reaction was partitioned between water and ethyl acetate, the aqueous phase was acidified to pH 2-3 with aqueous hydrochloric acid and extracted with ethyl acetate (3x300 mL). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep) ^TMCartridge silica) and gradient eluted with 0-20% methanol in dichloromethane afforded the desired product as a dark yellow gum (7.06g,17.3mmol, 62%).

LC/MS(C₂₄H₂₅NO₅)408[M+H]⁺；RT 0.74(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.90(t,J＝6.8Hz,2H),7.65(dd,J＝7.5,1.1Hz,2H),7.42(td,J＝7.4,3.0Hz,2H),7.34(td,J＝7.4,1.3Hz,2H),4.43-4.22(m,3H),3.50-3.42(m,1H),3.39-3.28(m,1H),3.26-3.15(m,3H),2.90-2.82(m,3H),2.51-2.44(m,2H),1.71(dt,J＝13.8,2.5Hz,3H)。

Step E(9H-fluoren-9-yl) methyl N- {2- [ (1-hydroxyhex-4-yn-2-yl) oxy ] oxy]Ethyl } -N-methylcarbamate

A solution of the product of step D (7.06g,17.33mmol,1eq) in tetrahydrofuran (120mL) was cooled to-10 deg.C, then a solution of triethylamine (2.65mL,19.1mmol,1.1eq) and isobutyl chloroformate (2.7mL,20.8mmol,1.2eq) in THF (40mL) was added dropwise. The precipitate was removed by filtration and the solution was cooled to-10 ℃. A solution of sodium borohydride (2.62g,69.3mmol,4eq) in water (40mL) was added dropwise and the mixture was stirred at-10 ℃ for 1 h. The pH of the solution was adjusted to pH 5 with 1N aqueous hydrochloric acid and then to pH 10 with saturated aqueous sodium bicarbonate. The layers were separated and the organic phase was washed successively with water (100mL) and brine (50mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,80 gReddiSep)^TMSilica gel cartridge) purification with a gradient of 0-100% ethyl acetate in isoheptaneElution afforded the desired product as a colorless gum (4.64g,11.8mmol, 68%).

LC/MS(C₂₄H₂₇NO₄)394[M+H]⁺；RT 0.77(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.90(d,J＝7.5Hz,2H),7.65(dt,J＝7.4,0.9Hz,2H),7.43(t,J＝7.4Hz,2H),7.35(td,J＝7.4,1.2Hz,2H),4.68-4.60(m,1H),4.39(d,J＝6.0Hz,1H),4.34(d,J＝6.7Hz,1H),4.28(t,J＝6.4Hz,1H),3.60-3.51(m,1H),3.46-3.36(m,2H),3.34-3.28(m,2H),3.19(dd,J＝16.6,5.5Hz,2H),2.84(d,J＝10.8Hz,3H),2.38-2.15(m,2H),1.71(t,J＝2.5Hz,3H)。

Step F(9H-fluoren-9-yl) methyl N- [2- ({1- [ (tert-butyldiphenylsilyl) oxy ]Hex-4-yn-2-yl } oxy) ethyl]-N-methylcarbamate

To a cooled solution of the product of step E (4.64g,11.8mmol,1eq) and imidazole (1.56mL,23.6mmol,2eq) in dichloromethane (200mL) was added tert-butyl (chloro) diphenylsilane (6.13mL,23.6mmol,2eq) dropwise and the mixture was warmed to room temperature and stirred overnight. The reaction was quenched with 2M aqueous ammonium chloride solution and the mixture was extracted with dichloromethane (3 × 200 mL). The combined organic extracts were washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-25% ethyl acetate in isoheptane to give the desired product as a colorless gum (5.86g,9.27mmol, 79%).

LC/MS(C₄₀H₄₅NO₄Si)632[M+H]⁺；RT 1.38(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.87(dd,J＝20.0,7.5Hz,2H),7.67-7.56(m,6H),7.53-7.39(m,7H),7.39-7.22(m,3H),4.38(t,J＝4.8Hz,1H),4.31(s,1H),4.24(t,J＝5.7Hz,1H),3.73-3.61(m,1H),3.60-3.44(m,2H),3.34-3.29(m,2H),3.29-3.18(m,1H),3.16-3.06(m,1H),2.81(d,J＝14.1Hz,3H),2.43-2.26(m,2H),1.69(t,J＝2.4Hz,3H),0.98(s,9H)。

Step G(9H-fluoren-9-yl) methyl N- [2- ({1- [ (tert-butyldiphenylsilyl) oxy]-3- (3, 6-dichloro-5-methylpyridazin-4-yl) propan-2-yl } oxy) ethyl]-N-methylCarbamic acid ester

The product of step F (5.86g,9.27mmol,1eq) and a solution of 3, 6-dichloro-1, 2,4, 5-tetrazine (5.6g,37.1mmol,4eq) in toluene (130mL) were heated in a sealed flask at 150 ℃ overnight. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,120g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-30% ethyl acetate in isoheptane to give the desired product as a pink foam (2.99g,3.97mmol, 43%).

LC/MS(C₄₂H₄₅Cl₂N₃O₄Si)754[M+H]⁺；RT 1.37(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.90(d,J＝7.7Hz,1H),7.78(d,J＝7.4Hz,1H),7.68-7.59(m,5H),7.57-7.50(m,1H),7.47-7.41(m,6H),7.45-7.37(m,1H),7.36-7.28(m,2H),7.23(t,J＝7.5Hz,1H),4.30(d,J＝5.7Hz,1H),4.27-4.11(m,2H),3.81-3.60(m,3H),3.55-3.45(m 1H),3.20-2.98(m,4H),2.89-2.77(m,1H),2.58(d,J＝23.0Hz,3H),2.39(d,J＝13.1Hz,3H),1.01(s,9H)。

Step H4- {3- [ (tert-butyldiphenylsilyl) oxy group]-2- [2- (methylamino) ethoxy]Propyl } -3, 6-dichloro-5-methylpyridazine

A solution of the product of step G (2.79G,3.7mmol,1eq) and diethylamine (0.77mL,7.39mmol,2eq) in acetonitrile (60mL) was stirred at room temperature overnight. Water was added and the mixture was extracted with ethyl acetate (3 × 70 mL). The combined organic extracts were washed with brine (100mL), dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-16% methanol in dichloromethane afforded the desired product as an orange/pink gum (1.9g,3.57mmol, 96%).

LC/MS(C₂₇H₃₅Cl₂N₃O₂Si)532[M+H]⁺；RT 0.84(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.69-7.62(m,4H),7.54-7.41(m,6H),3.83-3.60(m,3H),3.42-3.36(m,1H),3.16-2.97(m,3H),2.45(s,3H),2.39-2.23(m,2H),2.06(s,3H),1.02(s,9H)。

Step IN- [2- ({1- [ (tert-butyldiphenylsilyl) oxy group)]-3- (3, 6-dichloro-5-methylpyridazin-4-yl) propan-2-yl } oxy) ethyl]-N-methylcarbamic acid tert-butyl ester

To a solution of the product of step H (1.9g,3.57mmol,1eq) in dichloromethane (100mL) was added di-tert-butyl dicarbonate (1.53mL,7.14mmol,2eq) followed by triethylamine (1.99mL,14.3mmol,4eq) and the mixture was stirred at room temperature for 4 hours. The reaction was partitioned between dichloromethane and water, the aqueous phase was acidified to pH 4 and extracted with dichloromethane (3 × 80 mL). The combined organic extracts were washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,40g RediSep) ^TMSilica gel cartridge) and gradient eluted with 0-25% ethyl acetate in isoheptane to give the desired product as a colorless gum (1.83g,2.9mmol, 81%).

LC/MS(C₃₂H₄₃Cl₂N₃O₄Si)532[M-Boc+H]⁺；RT 1.33(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.69-7.62(m,4H),7.54-7.41(m,6H),3.76(qd,J＝10.7,4.7Hz,2H),3.66(d,J＝5.5Hz,1H),3.44(q,J＝7.9,6.3Hz,1H),3.20-3.10(m,3H),3.04(dd,J＝14.0,4.1Hz,2H),2.58(s,3H),2.44(s,3H),1.31(d,J＝22.6Hz,9H),1.02(s,9H)。

Step JN- (2- { [1- (3, 6-dichloro-5-methylpyridazin-4-yl) -3-hydroxypropan-2-yl]Oxy } ethyl) -N-methylcarbamic acid tert-butyl ester

A solution of the product of step I (1.83g,2.9mmol,1eq) in tetrahydrofuran (75mL) was cooled to 0 deg.C, then tetrabutylammonium fluoride (1M solution in tetrahydrofuran; 2.9mL,2.9mmol,1eq) was added and stirred at 0 deg.C for 30 minutes, then at room temperature for 1 hour. The reaction was partitioned between dichloromethane and water and the aqueous phase was extracted with dichloromethane (× 2). The combined organic extracts were washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,24g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a pale orange gum (0.73g,1.86mmol, 64%).

¹H NMR(400MHz,DMSO-d6)δ4.93(t,J＝5.5Hz,1H),3.62-3.44(m,4H),3.23(dt,J＝9.6,6.0Hz,1H),3.11(d,J＝23.9Hz,2H),3.02(dd,J＝6.5,2.0Hz,2H),2.60(d,J＝8.1Hz,3H),2.45(s,3H),1.35(d,J＝13.0Hz,9H)。

Step K2- { [ (tert-butoxy) carbonyl][2- (2- { [ (tert-butoxy) carbonyl ] carbonyl](methyl) amino } ethoxy) -3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl]Amino } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step J (125mg,0.32mmol,1eq) in toluene (20mL) was added the product of preparation 1c (171mg,0.35mmol,1.1eq), di-tert-butyl azodicarboxylate (146mg,0.63mmol,2eq) and triphenylphosphine (166mg,0.63mmol,2eq) and the mixture was stirred at 50 ℃ for 1 h. The reaction was partitioned between dichloromethane and water and the aqueous phase was extracted with dichloromethane (× 2), the combined organic extracts were washed with brine, dried (magnesium sulphate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-100% ethyl acetate in isoheptane to give the desired product as a pale yellow gum (282mg,0.32mmol, 102%).

LC/MS(C₄₀H₅₃Cl₂FN₆O₈S)867[M+H]⁺；RT 0.97(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.30(dd,1H),7.23-7.17(m,1H),7.12(t,1H),4.29(dd,J＝13.9,5.7Hz,1H),4.10(t,J＝6.0Hz,2H),3.96-3.87(m,1H),3.74(s,3H),3.61-3.48(m,1H),3.42(s,3H),3.32(s,2H),3.25(dt,J＝7.1,3.9Hz,3H),3.16-2.99(m,2H),2.97-2.89(m,1H),2.58(d,J＝11.6Hz,2H),2.45(s,3H),2.23(s,6H),2.10(t,J＝6.9Hz,2H),1.52(s,9H),1.31(d,J＝39.6Hz,9H)。

Step L2- { [2- (2- { [ (tert-butoxy) carbonyl](methyl) amino } ethoxy) -3- (3, 6-dichloro-5-methylpyridazin-4-yl) propyl]Amino } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

The product of step K (275mg,0.32mmol,1eq) was purified by 1,1,1,3,3, 3-hexafluoro-2-propanol (2.5mL,23.7 mmol)l,74.7eq) solution was heated for 60 minutes at 100 ℃ under microwave irradiation. The reaction solution was concentrated in vacuo and passed through an automatic flash column chromatography (CombiFlash Rf,12g RediSep) ^TMCartridge silica) and gradient eluted with 0-7% methanol in dichloromethane afforded the desired product as a white solid (154mg,0.2mmol, 63%).

LC/MS(C₃₅H₄₅Cl₂FN₆O₆S)767[M+H]⁺；RT 0.70(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.83(br s,1H),7.30(dd,J＝11.9,2.0Hz,1H),7.24-7.17(m,1H),7.12(t,J＝8.7Hz,1H),4.08(t,J＝6.1Hz,2H),3.82(dt,J＝9.0,4.5Hz,1H),3.70(s,3H),3.60-3.49(m,1H),3.46-3.39(m,4H),3.33(s,2H),3.29-3.18(m,1H),3.14(t,2H),3.10-3.02(m,2H),2.98(dd,J＝13.9,3.8Hz,1H),2.64-2.53(m,2H),2.44(s,3H),2.23(s,6H),2.07-1.95(m,2H),1.32(d,J＝30.8Hz,9H)。

Step M2- [6- (2- { [ (tert-butoxy) carbonyl](methyl) amino } ethoxy) -3-chloro-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl radicals]-5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step L (154mg,0.2mmol,1eq) in 1, 4-dioxane (14mL) were added cesium carbonate (131mg,0.4mmol,2eq), N-diisopropylethylamine (0.07mL,0.4mmol,2eq), and bis (di-tert-butyl (4-dimethylaminophenyl) phosphine) palladium (II) dichloride (14.2mg,0.02mmol,0.1eq) and the mixture was heated at 80 ℃ for 45 min. The reaction was partitioned between dichloromethane and water and the aqueous phase was extracted with dichloromethane (× 2). The combined organic extracts were washed with brine, dried (magnesium sulfate) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,12g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-8% methanol in dichloromethane afforded the desired product as a cream solid (136mg,0.19mmol, 93%).

LC/MS(C₃₅H₄₄ClFN₆O₆S)731[M+H]⁺；RT 0.75(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ7.31(dt,J＝12.0,1.9Hz,1H),7.25-7.19(m,1H),7.14(t,1H),4.86(dd,1H),4.25(s,1H),4.13(t,J＝6.2Hz,2H),3.93(d,J＝13.5Hz,1H),3.78(s,3H),3.56(t,J＝5.6Hz,2H),3.42(s,3H),3.32(s,2H),3.30-3.23(m,2H),3.21-3.09(m,2H),3.08-3.00(m,1H),2.58-2.52(m,1H),2.34(s,3H),2.23(s,6H),2.12(p,J＝6.7Hz,2H),1.27(d,J＝28.5Hz,9H)。

Step N2- {3- [ (1, 3-benzothiazol-2-yl) amino]-6- (2- { [ (tert-butoxy) carbonyl ](methyl) amino } ethoxy) -4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step M (136mg,0.19mmol,1eq) in cyclohexanol (4.5mL) was added 2-aminobenzothiazole (55.7mg,0.37mmol,2eq) and N, N-diisopropylethylamine (0.1mL,0.56mmol,3eq) and the mixture was purged with nitrogen (10 min). Xantphos (21.5mg,0.04mmol,0.2eq) and tris (dibenzylideneacetone) dipalladium (0) (17mg,0.02mmol,0.1eq) were added and the mixture was heated at 140 ℃ for 1 hour under microwave irradiation. The reaction was partitioned between dichloromethane and water and the aqueous phase was extracted with dichloromethane (3 × 40 mL). The combined organic extracts were washed with brine, dried (PTFE phase separator) and concentrated in vacuo. Purification by reverse phase automatic flash chromatography (CombiFlash Rf, c 1815.5 g Gold redissep column) eluting with a gradient of 5-95% acetonitrile in water afforded the desired product as a yellow solid (70.8mg,0.08mmol, 45%).

LC/MS(C₄₂H₄₉FN₈O₆S₂)845[M+H]⁺；RT 0.86(LCMS-V-B2)

¹H NMR(400MHz,DMSO-d6)δ11.52(br s,1H),7.88(d,J＝7.8Hz,1H),7.49(d,J＝8.1Hz,1H),7.37(ddd,J＝8.2,7.3,1.3Hz,1H),7.31(dd,J＝11.9,1.9Hz,1H),7.24-7.12(m,3H),4.80(dd,1H),4.22(s,1H),4.15(t,J＝6.2Hz,2H),3.94(d,J＝13.4Hz,1H),3.78(s,3H),3.56(t,J＝5.7Hz,2H),3.44-3.37(m,1H),3.31(s,2H),3.28(d,1H),3.24-3.14(m,2H),3.12-2.97(m,2H),2.58(d,J＝12.3Hz,3H),2.33(s,3H),2.19(s,6H),2.14(q,J＝7.0Hz,2H),1.27(d,9H)。

Step O2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6- [2- (methylamino) ethoxy]-5H,6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl) ]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of step N (70.8mg,0.08mmol,1eq) in dichloromethane (5mL) was added trifluoroacetic acid (1mL) slowly and the mixture was stirred at room temperature for 1 h. The reaction was partitioned between dichloromethane and saturated aqueous sodium bicarbonate and the aqueous phase was extracted with dichloromethane (3 × 30 mL). The combined organic extracts were washed with brine, dried (PTFE phase separator) and concentrated in vacuo to afford the desired product as a bright yellow solid (59.8mg,0.08mmol, 96%).

LC/MS(C₃₇H₄₁FN₈O₄S₂)745[M+H]⁺；RT 1.07(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.88(dd,J＝7.8,1.2Hz,1H),7.49(d,J＝8.1Hz,1H),7.37(ddd,J＝8.2,7.2,1.3Hz,1H),7.32(dd,J＝11.9,1.9Hz,1H),7.24-7.12(m,3H),4.79-4.69(m,1H),4.26-4.19(m,1H),4.15(t,J＝6.2Hz,2H),4.03(dd,J＝13.5,2.4Hz,1H),3.78(s,3H),3.60(t,J＝5.5Hz,2H),3.39(s,2H),3.32-3.27(m,2H),3.15(d,J＝14.6Hz,1H),3.08-2.99(m,1H),2.70(t,J＝5.5Hz,2H),2.38(s,3H),2.29(s,3H),2.22(s,6H),2.17-2.08(m,2H)。

Step P2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6- [2- (methylamino) ethoxy]-5H,6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl)]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step O (59.8mg,0.08mmol,1eq) in 1, 4-dioxane (2mL) was added 1M aqueous lithium hydroxide (0.24mL,0.24mmol,3eq) and the mixture was heated at 50 ℃ for 2 h. The solid was collected by filtration and dried in vacuo to give the desired product as a bright yellow solid (43mg,0.06mmol, 73%) as a lithium salt.

HRMS-ESI(m/z)[M+H]+C₃₆H₄₀FN₈O₄S₂731.2598, found 731.2623.

Example 66:2- {3- [ (1, 3-benzothiazol-2-yl) amino ]-4-methyl-5H, 6H,7H,8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ (6-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) oxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Step A5- {3- [ (6-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) oxy]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of 6-fluoro-2-methyl-3, 4-dihydro-1H-isoquinolin-7-ol (52mg,0.29mmol,1.8eq) and the product of preparation 3b (100mg,0.16mmol,1eq) in toluene (5mL) was added triphenylphosphine (83.7mg,0.32mmol,2eq) and di-tert-butyl azodicarboxylate (73.5mg,0.32mmol,2eq) and the mixture was stirred at 50 ℃ overnight. The reaction was partitioned between dichloromethane and water, the organic phase washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow solid (61mg,0.08mmol, 48%).

LC/MS(C₃₉H₄₈FN₇O₄SiS₂)790[M+H]⁺；RT 2.68(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(d,1H),7.48-7.37(m,2H),7.28-7.19(m,1H),6.94(d,J＝12.1Hz,1H),6.82(d,J＝8.7Hz,1H),5.84(s,2H),4.27(t,J＝5.8Hz,2H),4.05(t,J＝6.2Hz,2H),3.79(s,3H),3.76-3.68(m,2H),3.42-3.34(m,4H),3.27(t,2H),2.88(t,J＝6.2Hz,2H),2.69(d,J＝6.0Hz,2H),2.38(s,3H),2.26(s,3H),2.17-1.99(m,4H),0.95-0.87(m,2H),-.011(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c ]Pyridazin-8-yl } -5- {3- [ (6-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) oxy]Propyl } -1, 3-thiazole-4-carboxylic acid methyl ester

Will be described in detailA solution of the product of A (67mg,0.08mmol,1eq) in dichloromethane (4mL) was cooled to 0 deg.C, trifluoroacetic acid (1.95mL,25.4mmol,300eq) was added and the mixture was stirred at room temperature for 6 h. Dichloromethane (10mL) was added and the solution was cooled to 0 ℃, washed with aqueous ammonia, the organic phase was separated (phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-10% methanol in dichloromethane afforded the desired product as a yellow solid (39mg,0.06mmol, 70%).

LC/MS(C₃₃H₃₄FN₇O₃S₂)660[M+H]⁺；RT 2.01(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.89(s,1H),7.51(br s,1H),7.38(t,J＝7.5Hz,1H),7.20(t,J＝7.5Hz,1H),6.95(d,J＝12.1Hz,1H),6.83(d,J＝8.7Hz,1H),4.27(t,J＝5.7Hz,2H),4.06(t,J＝6.3Hz,2H),3.80(s,3H),3.43-3.36(m,4H),3.29(t,2H),2.89(t,J＝6.3Hz,2H),2.73-2.66(m,2H),2.35(s,3H),2.27(s,3H),2.17-2.10(m,2H),2.09-2.00(m,2H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ (6-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) oxy]Propyl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (39mg,0.06mmol,1eq) in 1, 4-dioxane (2mL) was added lithium hydroxide monohydrate (24.8mg,0.59mmol,10eq) and the mixture was heated at reflux overnight. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-20% 7N methanolic ammonia in dichloromethane afforded the desired product as an off white solid (19.4mg,0.03mmol, 51%).

HRMS-ESI(m/z)[M+H]+C₃₂H₃₃FN₇O₃S₂Calculated value of (2) 646.2070, found value of 646.2094

Example 675- [3- [4- [3- (azetidin-1-yl) propyl ] amide]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

Step A5- [3- [4- [3- (azetidin-1-yl) propyl ] amide]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 50mg of preparation 3e (0.077mmol,1.0eq.) as the appropriate alcohol and azetidine (88.00mg,20eq.) to give 35mg of the desired product (75%).

Step B5- [3- [4- [3- (azetidin-1-yl) propyl ] amide]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₄H₃₆FN₇O₃S₂674.2377, found 674.2386.

Example 682- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) propyl ] methyl ]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) propyl ] methyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 50mg of preparation 3e (0.077mmol,1.0eq.) as the appropriate alcohol and using 1-methylpiperazine (154.4mg,20eq.) to give 46mg of the desired product (73%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (4-methylpiperazin-1-yl) propyl ] methyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₄₂FN₈O₃S₂717.2799, found 717.2808.

Example 692- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylpropyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylpropyl) phenoxy ]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 50mg of preparation 3e (0.077mmol,1.0eq.) as the appropriate alcohol and pyrrolidine (109.6mg,20eq.) to give 53mg of the desired product (98%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-pyrrolidin-1-ylpropyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₅H₃₉FN₇O₃S₂688.2534, found 688.2533.

Example 702- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-morpholinopropyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-morpholinopropyl) phenoxy ] phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for preparation of propynylamine, starting with 50mg of preparation 3e (0.077mmol,1.0eq.) as the appropriate alcohol and morpholine (134.3mg,20eq.) gives 46mg of the desired product (83%).

Step B 2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-morpholinopropyl) phenoxy ] phenoxy]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₅H₃₉FN₇O₄S₂704.2483, found 704.2471.

Example 712- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (1-piperidinyl) propyl group]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (1-piperidinyl) propyl group]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 50mg of preparation 3e (0.077mmol,1.0eq.) as the appropriate alcohol and piperidine (131.2mg,20eq.) to give 43mg of the desired product (83%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (1-piperidinyl) propyl group]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₄₁FN₇O₃S₂702.2690, found 702.2703.

Example 722- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (1R,5S) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl]Propyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl]Propyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for preparation of propynylamine, starting with 50mg of preparation 3e (0.077mmol,1.0eq.) as the appropriate alcohol and using (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0] hexane (1.54mmol,20eq.) gives 24mg of the desired product (41%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (1R,5S) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl]Propyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₆H₃₇F₃N7O₃S₂736.2345, found 736.2340.

Example 732- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-oxo-2, 8-diazaspiro [4.5]]Decan-8-yl) propyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-oxo-2, 8-diazaspiro [4.5]]Decan-8-yl) propyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

The general procedure was prepared using propynylamine, starting with 50mg of preparation 3e (0.077mmol,1.0eq.) as the appropriate alcohol and using 2, 8-diazaspiro [4.5] decan-3-one (237.7mg,20eq.) to give 35mg of the desired product (58%).

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-oxo-2, 8-diazaspiro [4.5]]Decan-8-yl) propyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₃₉H₄₄FN₈O₄S₂771.2905, found 771.2922.

Example 742- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ (7-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) oxy ]Propyl } -1, 3-thiazole-4-carboxylic acid

Step A5- {3- [ (7-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) oxy]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethyl esterOxy radical]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazin-8-yl) -1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (120mg,0.19mmol,1eq) in toluene (5mL) was added 7-fluoro-2-methyl-3, 4-dihydro-1H-isoquinolin-6-ol (69.4mg,0.38mmol,2eq), triphenylphosphine (100mg,0.38mmol,2eq) and di-tert-butyl azodicarboxylate (88.2mg,0.38mmol,2 eq). The mixture was stirred at 50 ℃ overnight. The reaction was partitioned between dichloromethane and water, the organic phase washed with brine, dried (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-14% methanol in dichloromethane afforded the desired product as a yellow gum (111mg,0.14mmol, 73%).

LC/MS(C₃₉H₄₈FN₇O₄SiS₂)790[M+H]⁺；RT 2.64(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(dd,J＝7.6,1.2Hz,1H),7.48-7.38(m,2H),7.28-7.20(m,1H),6.94-6.84(m,2H),5.85(s,2H),4.27(t,J＝5.8Hz,2H),4.06(q,J＝7.4,6.8Hz,2H),3.79(s,3H),3.76-3.68(m,2H),3.34-3.22(m,4H),2.88(t,J＝6.3Hz,2H),2.73(t,J＝6.0Hz,2H),2.38(s,3H),2.27(s,3H),2.17-1.99(m,4H),0.97-0.86(m,2H),-0.12(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ (7-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) oxy]Propyl } -1, 3-thiazole-4-carboxylic acid methyl ester

A solution of the product of step A (111mg,0.14mmol,1eq) in dichloromethane (5mL) was cooled to 0 deg.C, trifluoroacetic acid (1.02mL,13.4mmol,95eq) was added and the mixture was stirred at room temperature overnight. Dichloromethane (10mL) was added and the solution was cooled to 0 ℃, washed with aqueous ammonia, the organic phase was separated (phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-7% methanol in dichloromethane afforded the desired product as a yellow gum (79mg,0.12mmol, 85%).

LC/MS(C₃₃H₃₄FN₇O₃S₂)660[M+H]⁺；RT 2.01(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.88(d,J＝7.8Hz,1H),7.50(br s,1H),7.42-7.33(m,1H),7.20(t,J＝7.7Hz,1H),6.95-6.85(m,2H),4.27(t,J＝5.7Hz,2H),4.08(t,J＝6.2Hz,2H),3.79(s,3H),3.34-3.22(m,6H),2.89(t,J＝6.3Hz,2H),2.74(t,J＝5.9Hz,2H),2.35(s,3H),2.28(s,3H),2.17-2.03(m,4H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [ (7-fluoro-2-methyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) oxy]Propyl } -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (79mg,0.12mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (50.2mg,1.2mmol,10eq) and the mixture was heated at reflux for 4.5 h. The reaction was cooled to room temperature and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-20% 7N methanolic ammonia in dichloromethane to give a solid which was triturated with ether, filtered, washed with ether and dried in vacuo to give the desired product as a yellow solid (44.4mg,0.07mmol, 57%).

HRMS-ESI(m/z)[M+H]+C₃₂H₃₃FN₇O₃S₂646.2070, found 646.2103.

Example 75:2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [4- (methylamino) butyl ] amino]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- {3- [4- (4- { [ (tert-butoxy) carbonyl](methyl) amino } butyl) -2-fluorophenoxy]Propyl } -2- (4-methyl-3- { [ (2Z) -3- { [2- (trimethylsilyl) ethoxy]Methyl } -2, 3-dihydro-1, 3-benzothiazol-2-ylidene]Amino } -5H,6H,7H, 8H-pyrido [2, 3-c)]Pyridazine-8-1, 3-thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 3b (50mg,0.08mmol,1eq) in toluene (5mL) was added the product of preparation 4k (32.7mg,0.11mmol,1.38eq), di-tert-butyl azodicarboxylate (36.7mg,0.16mmol,2eq) and triphenylphosphine (41.8mg,0.16mmol,2eq) and the mixture was heated at 50 ℃ overnight. The reaction was partitioned between dichloromethane and water, separated (PTFE phase separator) and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-70% ethyl acetate in isoheptane to give the desired product as a clear gum (45mg,0.05mmol, 62%).

LC/MS(C₄₅H₆₀FN₇O₆SiS₂)906[M+H]⁺；RT 3.51(LCMS-V-C)

¹H NMR(400MHz,DMSO-d6)δ7.82(d,1H),7.48-7.38(m,2H),7.24(ddd,J＝8.3,6.8,1.8Hz,1H),7.11-7.01(m,2H),6.91(d,J＝8.2Hz,1H),5.85(s,2H),4.27(t,J＝5.8Hz,2H),4.07(t,2H),3.78(s,3H),3.76-3.68(m,2H),3.32-23(m,4H),3.16-3.05(m,2H),2.88(t,J＝6.3Hz,2H),2.70(s,3H),2.38(s,3H),2.17-2.01(m,4H),1.51-1.39(m,4H),1.34(d,J＝16.8Hz,9H),0.95-0.86(m,2H),-0.11(s,9H)。

Step B2- {3- [ (1, 3-benzothiazol-2-yl) amino ]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [4- (methylamino) butyl]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid methyl ester

A solution of the product of step A (45mg,0.05mmol,1eq) in dichloromethane (3mL) was cooled to 0 deg.C, trifluoroacetic acid (0.61mL,7.95mmol,160eq) was added and the mixture was stirred at room temperature for 24 h. Dichloromethane (40mL) was added and the solution was washed with aqueous ammonia and concentrated in vacuo. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) and gradient eluted with 0-20% methanol in dichloromethane afforded the desired product as a yellow solid (22mg,0.03mmol, 66%).

LC/MS(C₃₄H₃₈FN₇O₃S₂)676[M+H]⁺；RT 1.12(LCMS-V-B1)

¹H NMR(400MHz,DMSO-d6)δ7.85(d,J＝7.9Hz,1H),7.47(d,J＝8.0Hz,1H),7.35(t,J＝7.5Hz,1H),7.16(t,J＝7.5Hz,1H),7.11-7.02(m,2H),6.93(d,J＝8.8Hz,1H),4.26(dd,J＝6.9,4.3Hz,2H),4.09(t,J＝6.1Hz,2H),3.79(s,3H),3.32-3.24(m,4H),2.88(t,J＝6.3Hz,2H),2.42(t,2H),2.34(s,3H),2.23(s,3H),2.19-2.09(m,2H),2.08-1.99(m,2H),1.60-1.47(m,2H),1.41-1.30(m,2H)。

Step C2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-5H, 6H,7H, 8H-pyrido [2,3-c]Pyridazin-8-yl } -5- (3- { 2-fluoro-4- [4- (methylamino) butyl]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

To a solution of the product of step B (22mg,0.03mmol,1eq) in 1, 4-dioxane (3mL) was added lithium hydroxide monohydrate (13.7mg,0.33mmol,10eq) and the mixture was heated at reflux overnight. By automatic flash column chromatography (CombiFlash Rf,4g RediSep)^TMSilica gel cartridge) was purified, eluting with a gradient of 0-25% 7N methanolic ammonia in dichloromethane to give a solid, which was suspended in ethyl acetate (1.5mL) and hydrochloric acid (4M in 1, 4-dioxane; 54.5. mu.L, 0.22mmol,6.7 eq). The mixture was stirred for 10 min, then the solid was collected by filtration and dried in vacuo to give the desired product as a yellow solid (11.4mg,0.02mmol, 53%) as the hydrochloride salt.

HRMS-ESI(m/z)[M+H]+C₃₃H₃₇FN₇O₃S₂662.2383, found 662.2414

Example 762- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [2- (dimethylamino) ethylamino ] amino]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A5- [3- [4- [3- [ tert-butoxycarbonyl- [2- (dimethylamino) ethyl ] amino]Amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

Using the Sonogashira general procedure starting from 1.00g of preparation 3a (1.66mmol,1eq.) and using 413mg of N- [2- (dimethylamino) ethyl ] -N-prop-2-ynyl-carbamic acid tert-butyl ester (1.83mmol,1.1eq.) as the appropriate alkyne, the desired product is isolated as a yellow solid.

¹H NMR(500MHz,DMSO-d₆)δppm 7.30(d,1H),7.21(d,1H),7.15(t,1H),4.27(brt,2H),4.26(t,2H),4.12(t,2H),3.77(s,3H),3.47(brt,2H),3.26(t,2H),2.89(t,2H),2.82(brs,2H),2.45(brs,6H),2.32(s,3H),2.11(qn,2H),2.04(qn,2H),1.43(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,155.4,151.8,151.4,151.4,147.5,142.4,136.2,135,129.1,129.1,119.2,115.5,114.8,82.3,80.3,68.3,56.3,52.0,46.4,46.4,44.6,43.1,30.7,28.5,24.2,23,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₃₄H₄₃ClFN₆O₅Calculated value of S is 701.2683, found value is 701.2678.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ tert-butoxycarbonyl- [2- (dimethylamino) ethyl ] ethyl]Amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure II, starting from the product of step A and 1, 3-benzothiazol-2-amine, the desired product was obtained. LC-MS-ESI (M/z) [ M + H ] ]⁺C₄₁H₄₈FN₈O₅S₂815.3, found 815.4.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [2- (dimethylamino) ethylamino ] amino]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure of deprotection and hydrolysis, followed by purification again by reverse phase preparative chromatography (C18,25mM NH)₄HCO₃MeCN) to obtain the desired product using the product of step B as a starting material.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₈FN₈O₃S₂Calculated value of (8): 701.2487, found 701.2483.

Example 772- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid

Step A5- [3- (2-fluoro-4-iodo-phenoxy) propyl]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the Mitsunobu general procedure, starting with 2.00g of preparation 3b (3.19mmol,1eq.) and 835mg of 2-fluoro-4-iodo-phenol (3.51mmol,1.1eq.) as the appropriate phenol, 2.31g (85% yield) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.81(dm,1H),7.6(dd,1H),7.45(dm,1H),7.43(dm,1H),7.41(m,1H),7.23(m,1H),7(t,1H),5.83(s,2H),4.25(t,2H),4.1(t,2H),3.77(s,3H),3.71(m,2H),3.26(t,2H),2.84(t,2H),2.34(s,3H),2.11(m,2H),2.03(m,2H),0.9(m,2H),-0.11(s,9H)；HRMS-ESI(m/z):[M+H]⁺C₃₅H₄₁FIN₆O₄S₂Calculated Si value 847.1423, found 847.1396.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- (2-fluoro-4-iodo-phenoxy) propyl]Thiazole-4-carboxylic acid

Using the general procedure of deprotection and hydrolysis, starting from the product of step a as the appropriate carbamate, the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₂₈H₂₅FIN₆O₃S₂703.0452, found 703.0427.

Example 782- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5-[3-[5-[3-(dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A5- [3- (2-fluoro-5-iodo-phenoxy) propyl]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the Mitsunobu general procedure, starting with 390mg of preparation 3b (0.622mmol,1eq.) and using 177mg 2-fluoro-5-iodo-phenol (0.746mmol,1.2eq.) as the appropriate phenol, 416mg (79% yield) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.81(dm,1H),7.46(dd,1H),7.43(dm,1H),7.41(m,1H),7.27(m,1H),7.23(m,1H),7.05(dd,1H),5.83(s,2H),4.26(t,2H),4.14(t,2H),3.78(s,3H),3.71(m,2H),3.26(t,2H),2.85(t,2H),2.34(s,3H),2.11(m,2H),2.04(m,2H),0.91(m,2H),-0.11(s,9H)；HRMS-ESI(m/z):[M+H]⁺C₂₈H₂₅FIN₆O₃S₂847.1423, found 847.1416.

Step B5- [3- [5- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene ]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the Sonogashira general procedure starting with 310mg of the product of step a (0.366mmol,1.0eq.) and 91mg of N, N-dimethylprop-2-yn-1-amine (1.10mmol,3eq.) as the appropriate alkyne gives 251mg (85% yield) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.81(dm,1H),7.43(dm,1H),7.41(m,1H),7.23(m,1H),7.23(m,1H),7.22(dd,1H),7.03(m,1H),5.82(s,2H),4.25(t,2H),4.15(t,2H),3.78(s,3H),3.71(m,2H),3.50(s,2H),3.27(t,2H),2.84(t,2H),2.33(s,3H),2.28(s,6H),2.12(m,2H),2.03(m,2H),0.9(m,2H),-0.11(s,9H)；HRMS-ESI(m/z):[M+H]⁺C₄₀H₄₉FN₇O₄S₂Calculated Si 802.3035, found 802.3028.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [5- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure of deprotection and hydrolysis, starting from the product of step B as the appropriate carbamate, the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₃₃H₃₃FN₇O₃S₂658.2064, found 658.2045.

Example 792- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-piperazin-1-ylprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (4-tert-butoxycarbonylpiperazin-1-yl) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical ]Thiazole-4-carboxylic acid

The general procedure for the preparation of propargylamine catalyzed by silver was used, starting from preparation 3c with paraformaldehyde as the aldehyde and piperazine-1-carboxylic acid tert-butyl ester as the appropriate secondary amine, to give the desired product.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-piperazin-1-ylprop-1-ynyl) phenoxy]Propyl radical]Thiazole-4-carboxylic acid

A mixture of the product of step a (207mg,0.25mmol) and HFxPyr (2.5mmol,10eq.) in acetonitrile (4.3mL) was stirred at 60 ℃ for 2.5 h. The product was purified by flash chromatography (24g silica gel column) using DCM and MeOH (NH)₃) As an eluent143mg (79%) of the desired product are obtained.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₆FN₈O₃S₂699.2330, found 699.2322.

Example 802- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (methylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A5- [3- (4-iodophenoxy) propyl group]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the general procedure of Mitsunobu, starting with 313mg of preparation 3b (0.50mmol,1.0eq.) and 110mg of 4-iodo-phenol (0.50mmol,1.0eq.) as the appropriate phenol, 328mg (63% yield) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.81(d,1H),7.58(d,2H),7.43(d,1H),7.42(t,1H),7.24(t,1H),6.81(d,2H),5.83(s,2H),4.25(t,2H),4.02(t,2H),3.78(s,3H),3.71(t,2H),3.26(t,2H),2.85(t,2H),2.35(s,3H),2.1(qn,2H),2.04(qn,2H),0.9(t,2H),-0.11(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 138.5,127.1,123.3,123.1,117.8,111.8,73,67.3,66.7,52.0,46.4,31.1,23.8,23.2,20.4,17.8,13.0,-0.9；HRMS-ESI(m/z):[M+H]⁺C₃₅H₄₂IN₆O₄S₂Calculated Si 829.1517, found 829.1517.

Step B5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group]Prop-1-ynyl]Phenoxy radical]Propyl radical]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the Sonogashira general procedure, starting with 3304mg of the product of step a (0.294mmol,1.0eq.) and using 100mg of N-methyl-N-prop-2-ynyl-carbamic acid tert-butyl ester (0.588mmol,2eq.) as the appropriate alkyne, 172mg (67% yield) of the desired product is obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.81(d,1H),7.44(d,1H),7.42(t,1H),7.36(d,2H),7.24(t,1H),6.96(d,2H),5.84(s,2H),4.26(t,2H),4.2(brs,2H),4.06(t,2H),3.78(s,3H),3.72(t,2H),3.28(t,2H),2.86(t,2H),2.84(brs,3H),2.36(s,3H),2.11(qn,2H),2.04(qn,2H),1.41(s,9H),0.91(t,2H),-0.11(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 133.5,127.1,123.3,123.1,115.3,111.8,72.9,67.3,66.7,52.0,46.3,38.6,33.7,31.0,28.5,23.8,23.2,20.3,17.8,13.0,-0.9；HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₆N₇O₆S₂Calculated Si value 870.3497, found 870.349.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (methylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

HRMS-ESI(m/z):[M+H]⁺C₃₂H₃₂N₇O₃S₂626.2002, found 626.2004.

Example 812- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- (3-methyl-3-pyrrolidin-1-yl-but-1-ynyl) phenoxy ]Propyl radical]Thiazole-4-carboxylic acid

The general procedure for the preparation of propargylamine catalyzed by silver was used, starting from preparation 3c, using acetone as the ketone and pyrrolidine as the appropriate secondary amine, to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₇H₃₉FN₇O₃S₂712.2534, found 712.2522.

Example 825- [3- [4- [3- (dimethylamino) prop-1-ynyl group]-2-fluoro-phenoxy]Propyl radical]-2- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

Step A2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- [4- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure of Sonogashira, starting with 500mg of preparation 3a (0.80mmol,1.0eq.) and using 100mg of N, N-dimethylprop-2-yn-1-amine (1.2mmol,1.5eq.) as the appropriate alkyne, 254mg (50% yield) of the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₂₇H₃₀ClFN₅O₃Calculated value of S is 558.1736, found value is 558.1729.

Step B5- [3- [4- [3- (dimethylamino) prop-1-ynyl group]-2-fluoro-phenoxy]Propyl radical]-2- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure II, using 254mg of the product of step A (0.45mmol,1.0eq.) and 153mg 7-fluoro-1, 3-benzothiazol-2-amine (0.91mmol,2.0eq.) as starting materials, 161mg (51% yield) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 11.59(brs,1H),7.41(dd,1H),7.4(t,1H),7.31(dd,1H),7.21(dd,1H),7.15(t,1H),7.08(t,1H),4.26(t,2H),4.13(t,2H),3.78(s,3H),3.49(s,2H),3.28(t,2H),2.87(t,2H),2.34(s,3H),2.27(s,6H),2.13(qn,2H),2.04(qn,2H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.2,157.0,155.7,151.6,150.4,149.1,148.8,147.5,141.6,134.9,129.0,128.3,128.0,127.9,119.3,117.2,115.5,115.0,113.6,108.4,84.8,84.3,68.3,51.8,48.0,46.4,43.9,30.8,23.9,22.9,20.2,12.8；HRMS-ESI(m/z):[M+H]+C₃₄H₃₄F₂N₇O₃S₂690.2127, found 690.2110.

Step C5- [3- [4- [3- (dimethylamino) prop-1-ynyl group]-2-fluoro-phenoxy]Propyl radical]-2- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

Using the general procedure of deprotection and hydrolysis, starting from the product of step B as the appropriate methyl ester, the desired product is obtained.

HRMS-ESI(m/z):[M+H]+C₃₂H₃₁FN₇O₃S₂676.1970, found 676.1958.

Example 832- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (dimethylamino) -3-methyl-but-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

The general procedure for the preparation of propargylamine catalyzed by silver was used, starting from preparation 3c, using acetone as the ketone and dimethylamine as the appropriate secondary amine, to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₇FN₇O₃S₂686.2377, found 686.2361.

Example 842- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [2- [1- (dimethylamino) cyclohexyl group]Ethynyl group]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

General procedure for the preparation of propargylamine using silver catalysis, starting from preparation 3c, using cyclohexanone as the ketone and dimethylamine as the appropriate secondary amine, gave the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₈H₄₁FN₇O₃S₂726.2690, found 726.2676.

Example 852- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (diethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

General procedure for the preparation of propargylamine using silver catalysis, starting from preparation 3c, using paraformaldehyde as the aldehyde and diethylamine as the appropriate secondary amine, gave the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₇FN₇O₃S₂686.2377, found 686.2386.

Example 862- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (diisopropylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

The general procedure for the preparation of propargylamine catalyzed by silver was used, starting from preparation 3c, using paraformaldehyde as the aldehyde and diisopropylamine as the appropriate secondary amine to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₇H₄₁FN₇O₃S₂714.2690, found 714.2681.

Example 87:2-[3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- [3- [4- [3- (diisobutylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

General procedure for the preparation of propargylamine catalyzed by silver using preparation 3c, paraformaldehyde as the aldehyde and N-isobutyl-2-methyl-propan-1-amine as the appropriate secondary amine, gives the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₉H₄₅FN₇O₃S₂742.3003, found 742.3001.

Example 882- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ ethyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

General procedure for the preparation of propargylamine using silver catalysis, starting from preparation 3c, using paraformaldehyde as the aldehyde and N-methylethylamine as the appropriate secondary amine, gives the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₄H₃₅FN₇O₃S₂672.2221, found 672.2206.

Example 892- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (3R,5S) -3, 5-dimethylpiperazin-1-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (3R,5S) -4-tert-butoxycarbonyl-3, 5-dimethyl-piperazin-1-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure for silver catalyzed propargylamine preparation, starting from preparation 3c, using paraformaldehyde as the aldehyde and tert-butyl (2R,6S) -2, 6-dimethylpiperazine-1-carboxylate as the appropriate secondary amine, 215mg (62% yield) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.88(dm,1H),7.49(brs,1H),7.37(m,1H),7.32(dd,1H),7.2(dm,1H),7.19(m,1H),7.15(t,1H),4.27(t,2H),4.14(t,2H),3.98(m,1H),3.49(s,2H),3.27(t,2H),2.88(t,2H),2.62/2.25(dd+dd,4H),2.34(s,3H),2.13(m,2H),2.04(m,2H),1.4(s,9H),1.19(d,6H)；HRMS-ESI(m/z):[M+H]⁺C₄₂H₄₈FN₈O₅S₂827.3167, found 827.3186.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ (3R,5S) -3, 5-dimethylpiperazin-1-yl]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

General procedure Using deprotection and hydrolysis (without LiOH x H)₂O hydrolysis), starting with the product of step a as the appropriate carbamate to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₇H₄₀FN₈O₃S₂727.2643, found 727.2641.

Example 902- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [1- [ (dimethylamino) methyl ] methyl]-3-bicyclo [1.1.1]Pentyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A5- [3- [4- [1- [ (dimethylamino) methyl group]-3-bicyclo [1.1.1]Pentyl radical]-2-fluoro-phenoxy]Propyl radical]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the general procedure of Mitsunobu, using preparation 4a as starting material and preparation 3b as appropriate alcohol, the crude desired product was isolated and transferred to the next step without further purification.

HRMS-ESI(m/z):[M+H]⁺C₄₃H₅₅FN₇O₄S₂Calculated Si value 844.3505, found 844.3485.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [1- [ (dimethylamino) methyl ] methyl]-3-bicyclo [1.1.1]Pentyl radical]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure of deprotection and hydrolysis, followed by purification again by reverse phase preparative chromatography (C18,25mM NH)₄HCO₃MeCN) to obtain the desired product using the product of step A as a starting material.

HRMS-ESI(m/z):[M+H]⁺C₃₆H₃₉FN₇O₃S₂700.2534, found 700.2515.

Example 912- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [ 3-methyl-3- (methylamino) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- [ 2-fluoro-4- [ 3-methyl-3- (methylamino) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure of Sonogashira, starting from preparation 3a and N, 2-dimethylbut-3-yn-2-amine, 417mg of the desired product are obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.23(dd,1H),7.16(dd,1H),7.12(t,1H),4.26(t,2H),4.11(t,2H),3.77(s,3H),3.25(t,2H),2.89(t,2H),2.37(s,3H),2.32(s,3H),2.1(m,2H),2.04(m,2H),1.34(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,151.3,136.2,129.1,128.9,119.1,115.4,93.7,81.5,68.2,52,51,46.4,30.7,30.4,29,24.2,23,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₂₈H₃₂ClFN₅O₃572.1898 as the calculated value of S; found 572.1888.

Step B2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ] ]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [ 3-methyl-3- (methylamino) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure II, starting from the product of step A and 1, 3-benzothiazol-2-amine, 77mg of the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₇FN₇O₃S₂686.2383; found value 686.2380

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [ 3-methyl-3- (methylamino) but-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure for hydrolysis, the product of step B was used as starting material to give 22mg of the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₄H₃₅FN₇O₃S₂672.2227; found 672.2224.

Example 926- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- (1- { [3- (2- { [ (3S) -3, 4-dihydroxybutyl)]Amino } ethoxy) -5, 7-dimethyladamantan-1-yl]Methyl } -5-methyl-1H-pyrazol-4-yl) pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using 2- [ (4S) -2, 2-dimethyl-1, 3-dioxolan-4-yl ] ethanamine as the appropriate amine, the compound having the dihydroxy-protected amine is obtained. Hydrolysis with 10% HCl solution (room temperature, 1 h) and purification by preparative HPLC using acetonitrile and 5mM aqueous ammonium bicarbonate as eluent gave the desired product.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₅N₉O₅822.4125, found 822.4120.

Example 936- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- (1- { [3- (2- { [ (3R) -3, 4-dihydroxybutyl)]Amino } ethoxy) -5, 7-dimethyladamantan-1-yl]Methyl } -5-methyl-1H-pyrazol-4-yl) pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using 2- [ (4R) -2, 2-dimethyl-1, 3-dioxolan-4-yl ] ethanamine as the appropriate amine, the compound having the dihydroxy-protected amine is obtained. Hydrolysis with 10% HCl solution (room temperature, 1 h) and purification by preparative HPLC using acetonitrile and 5mM aqueous ammonium bicarbonate as eluent gave the desired product.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₆N₉O₅Calculated value of S is 822.4125, found value is 822.4124.

Example 946- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- [1- ({3, 5-dimethyl-7- [2- (methylamino) ethoxy)]Adamantan-1-yl } methyl) -5-methyl-1H-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 12 and methylamine as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₁H₅₀N₉O₃Calculated value of S is 748.3757, found value is 748.3746.

Example 956- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- [1- ({3- [2- (dimethylamino) ethoxy)]-5, 7-dimethyladamantan-1-yl } methyl) -5-methyl-1H-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 12 and dimethylamine as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₂H₅₂N₉O₃Calculated value of S is 762.3914, found value is 762.3912.

Example 966- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- {1- [ (3, 5-dimethyl-7- {2- [ (2-sulfoethyl) amino group]Ethoxy } adamantan-1-yl) methyl]-5-methyl-1H-pyrazol-4-yl } pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation example 12 and taurine as suitable amine were used as starting materials and K was used in the substitution step₂CO₃(10eq) as base to give the desired product.

HRMS-ESI(m/z):[M+H]+C₄₂H₅₂N₉O₆S₂842.3482, found 842.3487.

Example 976- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- {1- [ (3, 5-dimethyl-7- {2- [ methyl (2-sulfoethyl) amino group]Ethoxy } adamantan-1-yl) methyl ]-5-methyl-1H-pyrazol-4-yl } pyridine-2-carboxylic acid

Example 982- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- {3- [4- (3- { [ (but-3-yn-1-yl) amino]Methyl bicyclo [1.1.1]Pentane-1-yl) -2-fluorophenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Example 996- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- [1- ({3- [2- (dimethylamino) ethoxy)]Adamantan-1-yl } methyl) -5-methyl-1H-pyrazol-4-yl]Pyridine-2-carboxylic acid

Example 1006- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -3- (1- { [3, 5-dimethyl-7- (4-methylpiperazin-1-yl) adamantan-1-yl)]Methyl } -5-methyl-1H-pyrazol-4-yl) pyridine-2-carboxylic acid

Example 1015- {3- [4- (3- { [ (3-azidopropyl) amino group]Methyl bicyclo [1.1.1]Pentane-1-yl) -2-fluorophenoxy]Propyl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1022- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- (3- {4- [3- (ethylamino) -3-methylbut-1-yn-1-yl-) ]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Step A5- [3- [4- [3- (tert-butoxycarbonylamino) -3-methyl-but-1-ynyl group]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

Using the general procedure of Sonogashira, starting from 1.00g of the product from preparation 3a (1.66mmol) and using 330mg (1.1eq) of tert-butyl N- (1, 1-dimethylprop-2-ynyl) carbamate as appropriate alkyne, 742mg (68%) of the desired product are obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.17(dd,1H),7.12(t,1H),7.11(dd,1H),7.07(brs,1H),4.26(t,2H),4.11(t,2H),3.77(s,3H),3.25(t,2H),2.89(t,2H),2.32(s,3H),2.1(qn,2H),2.04(qn,2H),1.50(s,6H),1.40(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,155.4,151.7,151.5,151.3,147.1,142.5,136.2,134.9,129.1,128.7,118.9,115.7,115.4,94.2,79.3,78.7,68.2,52.0,47.1,46.4,30.7,29.8,28.7,24.2,23.1,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₃₂H₃₈ClFN₅O₅Calculated value of S658.2266, found value of 658.2245

Step B5- [3- [4- (3-amino-3-methyl-but-1-ynyl) -2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

The product of step A (740mg,1.12 mmo)l) and HFxPyr (3eq) in acetonitrile (5mL/mmol) were stirred at 50 ℃ for 1 hour. After removal of volatiles, purification by column chromatography (silica gel, using EtOAc and MeOH (NH)₃) As eluent) to yield 560mg (89%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.18(dd,1H),7.11(m,1H),7.11(m,1H),4.25(m,2H),4.10(t,2H),3.77(s,3H),3.25(t,2H),2.88(t,2H),2.31(s,3H),2.10(m,2H),2.04(m,2H),1.35(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,128.6,118.9,115.3,98.1,78.4,68.2,52.0,46.3,46.3,32.3,30.7,24.2,23.1,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₂₇H₃₀ClFN₅O₃Calculated value of S is 558.1742, found value is 558.1730.

Step C2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- [4- [3- (ethylamino) -3-methyl-but-1-ynyl ]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

A mixture of the product of step B (550mg,0.98mmol), N-ethyl-N-isopropyl-propan-2-amine (0.52mL,3eq) and iodoethane (0.12mL,1.5eq) in N, N-dimethylformamide (5mL/mmol) was stirred at room temperature for 3 hours. After removal of volatiles, the crude intermediate was purified by column chromatography (silica gel, using EtOAc and MeOH (NH)₃) As eluent) to yield 570mg (99%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 9.07(brm,2H),7.41(dd,1H),7.29(dd,1H),7.20(t,1H),4.27(t,2H),4.15(t,2H),3.78(s,3H),3.27(t,2H),3.17(m,2H),2.90(t,2H),2.33(s,3H),2.12(qn,2H),2.05(qn,2H),1.64(s,6H),1.26(t,3H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,155.4,151.7,151.4,151.3,148.2,142.4,136.3,135.0,129.4,129.1,119.5,115.5,113.2,86.8,85.3,68.3,53.8,52.0,46.4,38.2,30.7,26.5,24.2,23.1,19.8,15.7,12.2；HRMS-ESI(m/z):[M+H]⁺C₂₉H₃₄ClFN₅O₃Calculated value of S is 586.2055, found value is 586.2048.

Step D2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (ethylamino) -3-methyl-but-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure I, starting from 570mg of the product of step C (0.98mmol) and 292mg (2eq) of 1, 3-benzothiazol-2-amine, 420mg (61%) of the desired product are obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 8.98(m,2H),7.88(brs,1H),7.53(brs,1H),7.42(dd,1H),7.38(m,1H),7.29(dm,1H),7.22(t,1H),7.20(m,1H),4.26(t,2H),4.16(t,2H),3.77(s,3H),3.29(t,2H),3.14(m,2H),2.88(t,2H),2.34(s,3H),2.15(m,2H),2.04(m,2H),1.60(s,6H),1.23(t,3H)；HRMS-ESI(m/z):[M+H]⁺C₃₆H₃₉FN₇O₃S₂700.2540, found 700.2532.

Step E2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (ethylamino) -3-methyl-but-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

To a mixture of the product of step D (420mg,0.60mmol) in a 2:1 mixture of 1, 4-dioxane and water (7.5mL/mmol) was added 50mg (2eq) of LiOH x H ₂O and the mixture was stirred at room temperature for 3 hours. After removal of volatiles, purification by reverse phase preparative chromatography (C18, 0.1% aqueous TFA: MeCN) afforded 33mg (8%) of the desired compound.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₇FN₇O₃S₂686.2383, found 686.2378.

Example 1032- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- (3- { 2-fluoro-4- [ 3-methyl-3- (piperazin-1-yl) but-1-yn-1-yl]Phenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Example 1045- [3- (4- {3- [ (3-azidopropyl) amino group]Prop-1-yn-1-yl } -2-fluoroPhenoxy) propyl group]-2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1055- [3- (4- {3- [ (3-aminopropyl) amino group)]Prop-1-yn-1-yl } -2-fluorophenoxy) propyl]-2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1062- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- [3- (2-fluoro-4- {3- [ (pent-4-yn-1-yl) amino]Prop-1-yn-1-yl } phenoxy) propyl]-1, 3-thiazole-4-carboxylic acid

Example 107 2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- {3- [ 2-fluoro-4- (3- { [2- (2-hydroxyethoxy) ethyl](methyl) amino } prop-1-yn-1-yl) phenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Example 1082- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- [3- (2-fluoro-4- { 3-methyl-3- [ (pent-4-yn-1-yl) amino]But-1-yn-1-yl } phenoxy) propyl]-1, 3-thiazole-4-carboxylic acid

Example 1092- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- {3- [ 2-fluoro-4- (3- { [ (prop-2-yn-1-yl) amino]Methyl bicyclo [1.1.1]Pentan-1-yl) phenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Example 1102- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- [3- (2-fluoro-4- {3- [ (hex-5-yn-1-yl) amino]Prop-1-yn-1-yl } phenoxy) propyl]-1, 3-thiazole-4-carboxylic acid

Example 1115- [3- (4- {3- [ (4-azidobutyl) amino group]Prop-1-yn-1-yl } -2-fluorophenoxy) propyl]-2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1125- [3- (4- {3- [ (4-azidobutyl) (methyl) amino group]Prop-1-yn-1-yl } -2-fluorophenoxy) propyl]-2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1132- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- [3- (2-fluoro-4- {3- [ (hex-5-yn-1-yl) (methyl) amino]Prop-1-yn-1-yl } phenoxy) propyl]-1, 3-thiazole-4-carboxylic acid

Example 1142- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- [3- (2-fluoro-4- {3- [ methyl (pent-4-yn-1-yl) amino]Prop-1-yn-1-yl } phenoxy) propyl]-1, 3-thiazole-4-carboxylic acid

Example 1155- [3- (4- {3- [ (3-azidopropyl) (methyl) amino group]Prop-1-yn-1-yl } -2-fluorophenoxy) propyl]-2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1162- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- {3- [ 2-fluoro-4- (3- { [ methyl (pent-4-yn-1-yl) amino]Methyl bicyclo [1.1.1]Pentan-1-yl) phenoxy ]Propyl } -1, 3-thiazole-4-carboxylic acid

Example 1175- {3- [4- (3- { [ (4-azidobutyl) (methyl) amino)]Methyl bicyclo [1.1.1]Pentane-1-yl) -2-fluorophenoxy]Propyl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1182- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- {3- [4- (3- { [ (but-3-yn-1-yl) (methyl) amino]Methyl bicyclo [1.1.1]Pentane-1-yl) -2-fluorophenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Example 1195- {3- [4- (3- { [ (3-azidopropyl) (methyl) amino]Methyl bicyclo [1.1.1]Pentane-1-yl) -2-fluorophenoxy]Propyl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1205- {3- [4- (3- { [ (4-azidobutyl) amino)]Methyl bicyclo [1.1.1]Pentane-1-yl) -2-fluorophenoxy]Propyl } -2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -1, 3-thiazole-4-carboxylic acid

Example 1212- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydropyrido [2,3-c ]]Pyridazin-8 (5H) -yl } -5- {3- [ 2-fluoro-4- (3- { [ (pent-4-yn-1-yl) amino ]Methyl } bicyclo [1.1.1]Pentan-1-yl) phenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Practice ofExample 1222- [ (6R) -3- [ (1, 3-benzothiazol-2-yl) amino]-6- (2-hydroxyethyl) -4-methyl-6, 7-dihydropyrido [2,3-c]Pyridazin-8 (5H) -yl]-5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Example 1232- [ (6S) -3- [ (1, 3-benzothiazol-2-yl) amino]-6- (2-hydroxyethyl) -4-methyl-6, 7-dihydropyrido [2,3-c]Pyridazin-8 (5H) -yl]-5- (3- {4- [3- (dimethylamino) prop-1-yn-1-yl]-2-fluorophenoxy } propyl) -1, 3-thiazole-4-carboxylic acid

Example 1242- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (prop-2-ynylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A5- [3- [ 2-fluoro-4- [3- (prop-2-ynylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]-2- [ 4-methyl-3- [ (Z) - [3- (2-trimethylsilylethoxymethyl) -1, 3-benzothiazol-2-ylidene]Amino group]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using the Sonogashira general procedure, starting from the product of example 77, step a (2.30g,2.71mmol,1.0eq.) and using 1.26g N-prop-2-ynylprop-2-yn-1-amine (13.58mmol,5eq.) as the appropriate alkyne, 793mg (36%) of the desired product was obtained.

LC/MS(C₄₁H₄₇FN₇O₄S₂Si)812[M+H]⁺。

Step B2- [3- (1, 3-benzothiazol-2-yl)Amino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (prop-2-ynylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Using the general procedure for deprotection and hydrolysis, the product of step A (900mg,1.10mmol) was used as starting material to give 222mg (30%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.88(d,1H),7.48(br.,1H),7.37(t,1H),7.28(dd,1H),7.19(d,1H),7.18(t,1H),7.14(t,1H),4.27(br.,2H),4.14(t,2H),3.55(s,2H),3.39(d,2H),3.27(t,2H),3.09(t,1H),2.87(t,2H),2.33(s,3H),2.13(m,2H),2.03(m,2H)；¹³C NMR(125MHz,DMSO-d₆)δppm 128.9,126.5,122.5,122.3,119.2,115.5,87.8,82.8,82.2,74.5,68.5,46.3,37.2,36.6,31.0,23.9,23.1,20.3,12.9；HRMS-ESI(m/z):[M+H]⁺C₃₄H₃₁FN₇O₃S₂668.1914, found 668.1907.

Example 125:2- {3- [ (1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H, -pyrido [2,3-c]Pyridazin-8-yl } -5- {3- [4- (3- { bis [ (3S) -3, 4-dihydroxybutyl) S]Amino } prop-1-yn-1-yl) -2-fluorophenoxy]Propyl } -1, 3-thiazole-4-carboxylic acid

Step A5- [3- [4- [3- (tert-butoxycarbonylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

Using the Sonogashira general procedure, starting with 3.00g of preparation 3a (5.0mmol) and using tert-butyl 1.55g N-prop-2-ynylcarbamate (2eq.) as the appropriate alkyne, 2.79g of the desired product are obtained (89%).

¹H NMR(500MHz,dmso-d6)δppm 7.34(brt,1H),7.26(dd,1H),7.17(dm,1H),7.13(t,1H),4.25(t,2H),4.11(t,2H),3.95(brd,2H),3.77(s,3H),3.25(t,2H),2.88(m,2H),2.31(s,3H),2.10(m,2H),2.03(m,2H),1.39(s,9H)；¹³C NMR(500MHz,dmso-d6)δppm 128.9,119.1,115.4,68.2,51.9,46.3,30.7,30.5,28.7,24.1,23.0,19.7,15.7；HRMS-ESI(m/z):[M+H]+C₃₀H₃₄ClFN₅O₅Calculated value of S is 630.1953, found value is 630.1945.

Step B5- [3- [4- (3-aminoprop-1-ynyl) -2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c) ]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

A mixture of the product of step A (2.19g,3.47mmol), pyridine and hydrogen fluoride (1:1) (3.44g,10.0eq) in MeCN (17.3mL) was stirred at 60 ℃ for 1.5 h. Purification by flash chromatography (silica gel, DCM and MeOH (1.2% NH)₃) As eluent) gave the desired product (1.81g, 98.5%).

¹H NMR(500MHz,dmso-d6)δppm 7.26(dd,1H),7.18(dd,1H),7.14(t,1H),5.36(NH3+,br.,3H),4.25(m,2H),4.12(t,2H),3.77(s,3H),3.61(s,2H),3.25(t,2H),2.88(t,2H),2.31(s,3H),2.10(m,2H),2.04(m,2H)；¹³C NMR(500MHz,dmso-d6)δppm 163.1,155.3,151.7,151.5,151.3,147.4,142.5,136.1,136.1,134.9,128.9,119.1,115.5,115.2,89.2,81.9,68.2,51.9,46.3,31.1,30.7,24.2,23.0,19.7,15.7；HRMS-ESI(m/z):[M+H]+C₂₅H₂₆ClFN₅O₃Calculated value of S is 530.1429, found value is 530.1410.

Step C(4S) -4- (2-iodoethyl) -2, 2-dimethyl-1, 3-dioxolane

To PPh₃(11.84g,2.2eq), imidazole (3.07g,2.2eq) and 2- [ (4S) -2, 2-dimethyl-1, 3-dioxolan-4-yl]A mixture of ethanol (2.92mL,20.52mmol) in dichloromethane (103mL) was added in portions to iodine (11.46g,2.2eq) at 0 deg.C, then stirred at room temperature for 18 h. Then using 100mL of Na₂S₂O₃The reaction was quenched and the phases separated, the organic phase washed with brine, dried and purified by flash chromatography (silica gel, heptane and heptane-MTBE as eluents) to give the desired compound (2.90g, 55%).

¹H NMR(500MHz,dmso-d6)δppm 4.06(m,1H),4.01/3.45(dd+dd,2H),3.28/3.21(dd+dd,2H),2.00/1.97(m+m,2H),1.31(s,3H),1.26(s,3H)；¹³C NMR(500MHz,dmso-d6)δppm108.7,75.8,68.3,37.9,27.3,26.0,3.5；GC-MS(EI,M+):255.79。

Step D5- [3- [4- [3- [ bis [2- [ (4S) -2, 2-dimethyl-1, 3-dioxolyl-4-yl)]Ethyl radical]Amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

A mixture of the product of step B (500mg,0.94mmol), the product of step C (483.2mg,2.0eq) and N-ethyl-N-isopropyl-propan-2-amine (1.0mL,6eq) in N, N-dimethylformamide (4.7mL) was stirred at room temperature for 6 hours. 10mL of a 2M dimethylamine solution was added and the reaction mixture was stirred for an additional 1 hour. The mixture was washed with water and saturated NaHCO₃The solution was diluted and extracted with EtOAc. The combined organic phases were washed with brine, dried, concentrated and purified by preparative HPLC (MeCN, NH)₄HCO₃) The desired compound (100mg, 13%) was obtained.

¹H NMR(500MHz,dmso-d6)δppm 7.28(dd,1H),7.18(dm,1H),7.14(t,1H),4.27(t,2H),4.11(t,2H),4.07(m,2H),3.99/3.46(dd+dd,4H),3.76(s,3H),3.56(s,2H),2.89(t,2H),2.51(m,4H),2.33(s,3H),2.32(t,2H),2.11(m,2H),2.04(m,2H),1.64(m,4H),1.30(s,6H),1.24(s,6H)；HRMS-ESI(m/z):[M+H]+C₃₉H₅₀ClFN₅O₇Calculated value of S is 786.3104, found value is 786.3111.

Step E2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ bis [2- [ (4S) -2, 2-dimethyl-1, 3-dioxol-4-yl]Ethyl radical]Amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure I, the starting material was used 100mg of the product of step D (0.127mmol) and purified by preparative HPLC (MeCN, NH)₄HCO₃) 90mg (78%) of the desired product are obtained.

¹H NMR(500MHz,dmso-d6)δppm 7.90(brs,1H),7.60(brs,1H),7.37(brm,1H),7.29(dd,1H),7.20(brm,1H),7.19(dm,1H),7.16(t,1H),4.45/3.98(dd+dd,4H),4.26(t,2H),4.14(t,2H),4.05(dd,2H),3.78(s,3H),3.53(s,2H),3.28(t,2H),2.88(t,2H),2.5(m,4H),2.35(s,3H),2.14(m,2H),2.05(m,2H),1.63(m,4H),1.3(s,6H),1.24(s,6H)；HRMS-ESI(m/z):[M+H]+C₄₆H₅₅FN₇O₇S₂900.3588, found 900.3591.

Step F2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals ]-5- [3- [4- [3- [ bis [ (3S) -3, 4-dihydroxybutyl ] butyl]Amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

The product of step E (90mg,0.1mmol) and LiOH x H₂A mixture of O (95mg,22.6eq) in 1, 4-dioxane (1mL) and water (1mL) was stirred at room temperature for 1 hour and at 50 ℃ for 2 hours. Treated with hydrogen chloride (8mmol) and stirred at room temperature for 4 h, then saturated NaHCO was added₃A 1:1 mixture of solution and water and brine and the desired product was filtered off (33mg, 40%).

HRMS-ESI(m/z)[M+H]⁺C₃₉H₄₅FN₇O₇S₂806.2806, found 806.2803.

Example 126:5- [3- [4- (3-aminoprop-1-ynyl) -2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Step A:2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- [3- [4- [3- (tert-butoxycarbonylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure I, starting from 4.60g of example 125, step A and 2.20g (2eq) of 2-aminobenzothiazole, and purification by column chromatography (silica gel using heptane, EtOAc and MeOH (1.2% NH3) as eluent), 4.02g (74%) of the desired product were obtained.

¹H NMR(500MHz,dmso-d6)δppm 7.90(br.,1H),7.61(br.,1H),7.37(brt.,1H),7.27(dd,1H),7.19(br.,1H),7.19(dd,1H),7.15(t,1H),4.25(t,2H),4.14(t,2H),3.94(d,2H),3.77(s,3H),3.27(t,2H),2.86(t,2H),2.33(s,3H),2.13(m,2H),2.04(m,2H),1.39(s,9H)；¹³C NMR(500MHz,dmso-d6)δppm 163.2,155.7,129.0,126.4,122.5,122.2,119.2,115.5,68.4,51.9,46.3,31.0,30.5,28.7,23.9,23.1,20.3,12.9；HRMS-ESI(m/z):[M+H]+C₃₇H₃₉FN₇O₅S₂744.2438, found 744.2425.

And B:5- [3- [4- (3-aminoprop-1-ynyl) -2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

A mixture of the product of step A (4.00g,5.38mmol), pyridine and hydrogen fluoride (1:1) (5.33g,10eq) in MeCN (27mL) was stirred at 60 ℃ for 16 h. Purification by column chromatography (silica gel, DCM and MeOH (NH)₃) As eluent) to yield the desired compound.

HRMS-ESI(m/z):[M+H]+C₃₂H₃₁FN₇O₃S₂644.1914, found 644.1913.

Example 127:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 12 and pyrrolidine as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₄N₉O₃Calculated value of S is 788,4070, found value is 788.4068.

Example 128:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl)Radical-7- [2- (4-methylpiperazin-1-yl) ethoxy ]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and 1-methylpiperazine as the appropriate amine, the desired product is obtained.

HRMS-ESI(m/z):[M+2H]²⁺C₄₅H₅₈N₁₀O₃Calculated value of S is 409.2207, found value is 409.2208.

Example 129:2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ [ (3S) -3, 4-dihydroxybutyl)]Amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

A mixture of the product of preparation 3c (400mg,0.67mmol), paraformaldehyde (400mg,20eq), (2S) -4-aminobutane-1, 2-diol hydrochloride (1:1) (754.3mg,8eq), triethylamine (2.3mL,25eq), CuI (127mg,1eq) and molecular sieve (0.5g) in ethanol (3.3mL) was held in an Anton-Paar microwave reactor at 120 ℃ for 1 hour. Purification by column chromatography (silica gel using heptane, EtOAc and MeOH/NH3(0.6N) as eluent) and RF HPLC (Gemini using water and 0.1% TFA and acetonitrile as eluents) gave 15.3mg (3%) of the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₇FN₇O₅S₂718.2282, found 718.2266.

Example 130:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c) ]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ [ (3R) -3, 4-dihydroxybutyl)]-methyl-amino]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using 2- [ (4R) -2, 2-dimethyl-1, 3-dioxolan-4-yl ] -N-methyl-ethylamine as the appropriate amine, the compound having the dihydroxy protected amine was obtained. Hydrolysis with 10% HCl solution (room temperature, 1 h) and purification by preparative HPLC using acetonitrile and 5mM aqueous ammonium bicarbonate as eluent gave the desired product.

HRMS-ESI(m/z):[M+2H]²⁺C₄₅H₅₉N₉O₅Calculated value of S is 418.7180, found value is 418.7167.

Example 131:2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ [ (3R) -3, 4-dihydroxybutyl)]Amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

A mixture of the product of preparation 3c (200mg,0.33mmol), paraformaldehyde (200mg,20eq), (2R) -4-aminobutane-1, 2-diol hydrochloride (1:1) (471mg,10eq), triethylamine (1.2mL,25eq), CuI (64mg,1eq) and molecular sieve (0.25g) in ethanol (1.6mL) was held in an Anton-Paar microwave reactor at 120 ℃ for 1 hour. Purification by column chromatography (silica gel, using heptane, EtOAc and MeOH/NH) ₃(0.6N) as eluent) and RF HPLC purification (Gemini, using water and 0.1% TFA and acetonitrile as eluents) gave 43mg (18%) of the desired product.

HRMS-ESI(m/z):[M+H]⁺C₃₅H₃₇FN₇O₅S₂718.2282, found 718.2281.

Example 132:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3-[1-[[3-[2-(4-Hydroxybutylamino) ethoxy]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using 4-aminobutan-1-ol as the appropriate amine, the desired product is obtained.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₆N₉O₄Calculated value of S is 806.4176, found value is 806.4174.

Example 133:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] group]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using (2, 2-dimethyl-1, 3-dioxan-5-yl) methylamine as the appropriate amine, the compound having the dihydroxy-protected amine is obtained. Hydrolysis with 10% HCl solution (room temperature, 1 h) and purification by preparative HPLC using acetonitrile and 5mM aqueous ammonium bicarbonate as eluent gave the desired product.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₆N₉O₅Calculated value of S822,4125, found value 822.4099.

Example 134:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ bis (2-hydroxyethyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 12 was used starting with 2- (2-hydroxyethylamino) ethanol as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₆N₉O₅Calculated value of S is 822,4125, found value is 822.4123.

Example 135:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ [ 2-hydroxy-1- (hydroxymethyl) ethyl ] ethyl]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using 2-aminopropane-1, 3-diol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₃H₅₄N₉O₅Calculated value of S is 808.3969, found value is 808.3965.

Example 136:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [2- (2-hydroxyethoxy) ethylamino ] amino ]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 12 was used starting with 2- (2-aminoethoxy) ethanol as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₆N₉O₅Calculated value of S is 822.4125, found value is 822.4116.

Example 137:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ bis (3-hydroxypropyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using 3- (3-hydroxypropylamino) propan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]+C₄₆H₆₀N₉O₅Calculated value of S is 850.4438, found value is 850.4436.

Example 138:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- (3-hydroxypropylamino) ethoxy ] ethyl]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and using 3-aminopropan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₃H₅₄N₉O₄Calculated value of S792.4019, found value 792.4012.

Example 139:5- [3- [4- [3- [ acetyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

Step A5- [3- [4- [3- [ tert-butoxycarbonyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

Using the general procedure of Sonogashira starting from 4.00g of preparation 3a (6.63mmol) and using 2.24g N-methyl-N-prop-2-ynyl-carbamic acid tert-butyl ester (13.3mmol,2eq) as the appropriate alkyne, 2.40g (55%) of the desired product are obtained.

¹H NMR(500MHz,dmso-d6)δppm 7.30(dd,1H),7.20(dm,1H),7.13(t,1H),4.24(m,2H),4.23(brs,2H),4.11(t,2H),3.77(s,3H),3.24(t,2H),2.87(m,2H),2.86(s,3H),2.30(s,3H),2.10(m,2H),2.03(m,2H),1.41(s,9H)；¹³C NMR(500MHz,dmso-d6)δppm 129.1,119.3,115.4,85.2,82.4,68.2,51.9,46.3,38.6,33.8,30.7,28.5,24.1,23.0,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₃₁H₃₆ClFN₅O₅Calculated value of S is 644.2110, found value is 644.2094.

Step B2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- [3- [ 2-fluoro-4- [3- (methylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

To a solution of 322mg of the product of step A (0.5mmol) in 2.5mL of acetonitrile was added 0.9mL of hydrogen fluoride in pyridine (20 eq). The reaction mixture was stirred at 60 ℃ until no further conversion was observed. Purification by flash chromatography (silica gel, using DCM and MeOH (1.2% NH) ₃) As eluent) gave 258mg (95%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 7.25(dd,1H),7.17(dd,1H),7.12(t,1H),4.25(t,2H),4.11(t,2H),3.77(s,3H),3.46(s,2H),3.25(t,2H),2.88(t,2H),2.32(s,3H),2.31(s,3H),2.10(qn,2H),2.03(qn,2H),1.99(brs,1H)；¹³C NMR(500MHz,dmso-d6)δppm 163.1,155.4,151.7,151.6,151.3,147.2,142.5,136.2,134.9,129.0,128.8,119.1,115.7,115.4,88.7,82.1,68.3,52.0,46.4,40.5,35.4,30.8,24.2,23.1,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₂₆H₂₈ClFN₅O₃Calculated value of S544.1585, found value 544.1570.

Step C5- [3- [4- [3- [ acetyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

To a solution of 220mg of the product of step B (0.41mmol) and 0.085mL of TEA (1.5eq) in 2mL of dichloromethane was added 0.031mL of acetyl chloride (1.1 eq). The reaction mixture was stirred until no further conversion was observed. Purification by flash chromatography (silica gel, using DCM and MeOH (1.2% NH)₃) As eluent) gave 174mg (73%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 7.34/7.31(dd/dd,1H),7.23/7.20(brd/brd.,1H),7.14/7.13(t/t,1H),4.38/4.34(s/s,2H),4.25(m,2H),4.12(t,2H),3.77(s,3H),3.25(t,2H),3.05/2.88(s/s,3H),2.88(t,2H),2.31(s,3H),2.10(m,2H),2.09/2.02(s/s,3H),2.03(m,2H)；¹³C NMR(500MHz,dmso-d6)δppm 170.2/170.1,163.0,155.4,151.3,142.4,134.9,129.2/129.1,119.4/119.3,115.4,85.3/84.7,82.9/81.9,68.2,51.9,46.3,40.6/36.3,35.4/33.1,30.7,24.1,23.0,21.9/21.8,19.7,15.7.HRMS-ESI(m/z):[M+H]⁺C₂₈H₃₀ClFN₅O₃Calculated value of S is 586.1691, found value is 586.1690.

Step D5- [3- [4- [3- [ acetyl (methyl) amino group]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure I, starting from 170mg (0.29mmol) of the product from step C and 87mg (2eq) of 1, 3-benzothiazol-2-amine, 220mg (98%) of the desired product are obtained.

HRMS-ESI(m/z):[M+H]+C₃₅H₃₅FN₇O₄S₂700.2176, found 700.2180.

Step E5- [3- [4- [3- [ acetyl (methyl) amino group ]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

The general procedure for hydrolysis was used with the starting material using the product of step D as the appropriate methyl ester to give the desired product.

HRMS-ESI(m/z):[M+H]+C₃₄H₃₃FN₇O₄S₂686.2023, found 686.2019.

Example 140:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ bis (4-hydroxybutyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 12 was used starting with 4- (4-hydroxybutylamino) butan-1-ol as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]+C₄₈H₆₄N₉O₅Calculated value of S is 878.4751, found value is 878.4752.

Example 141:2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- [3- [4- [3- (dimethylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A4- [3- (dimethylamino) prop-1-ynyl]Phenol and its preparation

Using the general procedure of Sonogashira, starting with 10.0g of 4-iodophenol (45.45mmol) and 4.91g (1.3eq) of N, N-dimethylprop-2-yn-1-amine, 3.29g (41%) of the desired product were obtained.

¹H NMR(400MHz,DMSO-d₆)δppm 9.83(brs,1H),7.25(d,2H),6.74(d,2H),3.44(s,2H),2.26(s,6H)；LC/MS(C₁₁H₁₄NO)176[M+H]⁺。

Step B:2-(tert-Butoxycarbonylamino) -5- [3- [ tert-butyl (diphenyl) silyl]Oxopropyl radical]Thiazole-4-carboxylic acid methyl ester

To a solution of the product of preparation 1a, step C (77.0g,243.7mmol), imidazole (33.14g,2eq) and DMAP (1.49g,0.05eq) in DMF (973mL) was added tert-butyl (chloro) diphenylsilane (93.5mL,1.5eq) dropwise and the reaction mixture was stirred at room temperature for 16 h. After removal of volatiles, purification by column chromatography (silica gel using heptane and EtOAc as eluent) gave 13.56g (99%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 11.63(s,1H),7.60(d,4H),7.45(t,2H),7.42(t,4H),3.74(s,3H),3.67(t,2H),3.20(t,2H),1.87(qn,2H),1.47(s,9H),0.99(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 162.8,156.0,142.6,135.6,135.5,133.5,130.3,128.3,81.8,62.9,51.9,34.0,28.3,27.1,23.2,19.2；HRMS-ESI(m/z):[M+H]⁺C₂₉H₃₉N₂O₅Calculated value of SSi 555.2349, found value 555.2336.

Step C2- [ tert-Butoxycarbonyl- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propyl ] oxy]Amino group]-5- [3- [ tert-butyl (diphenyl) silyl]Oxopropyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for alkylation starting from 34.95g (63mmol) of the product from step B and using 25.0g (1.2eq) of 3, 6-dichloro-4- (3-iodopropyl) -5-methyl-pyridazine as the appropriate iodide compound, 51.0g (quantitative yield) of the desired product was obtained.

¹H NMR(500MHz,DMSO-d₆)δppm 7.63-7.37(m,10H),4.09(t,2H),3.75(s,3H),3.67(t,2H),3.20(t,2H),2.82(m,2H),2.40(s,3H),1.87(m,2H),1.87(m,2H),1.50(s,9H),0.97(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 62.9,52.0,46.1,33.9,28.1,27.5,27.1,25.9,23.8,16.4；HRMS-ESI(m/z):[M+H]⁺C₃₇H₄₇C_l2N₄O₅Calculated value of SSi 757.2413, found value 757.2395.

Step D5- [3- [ tert-butyl (diphenyl) silyl]Oxopropyl radical]-2- [3- (3, 6-dichloro-5-methyl-pyridazin-4-yl) propylamino]Thiazole-4-carboxylic acid methyl ester

Using the general procedure for deprotection with HFIP, 51.70g of the product of step C (68mmol) were used as starting material to give 36.32g (81%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.71(t,1H),7.63-7.37(m,10H),3.69(s,3H),3.67(t,2H),3.30(m,2H),3.10(t,2H),2.85(m,2H),2.83(s,3H),1.79(m,2H),1.78(m,2H),0.98(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 62.9,51.7,44.1,34.2,28.0,27.1,27.0,23.4,16.4；HRMS-ESI(m/z):[M+H]⁺C₃₂H₃₉Cl₂N₄O₃Calculated value of SSi 657.1889, found value 657.1875.

Step E5- [3- [ tert-butyl (diphenyl) silyl]Oxopropyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid methyl ester

36.0g (54.7mmol) of the product of step D and 35.7g (2eq) of Cs₂CO₃The mixture in 1, 4-dioxane (383mL) was stirred at 90 ℃ for 18 h. After dilution with water, the precipitated solid was filtered off, washed with diethyl ether and dried to yield 34.0g (99%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.61(d,4H),7.43(t,2H),7.42(t,4H),4.26(t,2H),3.77(s,3H),3.70(t,2H),3.23(t,2H),2.90(t,2H),2.33(s,3H),2.04(qn,2H),1.90(qn,2H),1.00(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,155.3,151.8,151.4,143.2,136.2,135.5,134.7,133.6,130.3,129.0,128.3,63.1,51.9,46.3,34.1,27.1,24.2,23.1,19.8,19.2,15.7；HRMS-ESI(m/z):[M+H]⁺C₃₂H₃₈ClN₄O₃Calculated value of SSi 621.2122, found value 621.2097.

Step F2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) -5- (3-hydroxypropyl) thiazole-4-carboxylic acid methyl ester

A mixture of 23.36g (37.6mmol) of the product of step E and 45mL (1.2eq.) of a 1M solution of TBAF in THF (5mL/mmol) was stirred at room temperature for 2 h. After removal of volatiles, purification by column chromatography (silica gel, using EtOAc and MeOH/NH)₃As eluent) to obtainTo 12.88g (89%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 4.54(br.,1H),4.25(m,2H),3.80(s,3H),3.45(t,2H),3.11(m,2H),2.88(t,2H),2.31(s,3H),2.04(m,2H),1.77(m,2H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,155.2,151.2,143.8,136.1,134.5,129.0,60.5,52.0,46.3,34.6,24.2,23.2,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₁₆H₂₀ClN₄O₃Calculated value of S is 383.0945, found value is 383.0937.

Step G2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c) ]Pyridazin-8-yl) -5- [3- [4- [3- (dimethylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using the general method of Mitsunobu, starting with 0.65g (1.2eq) of the product of step F and 250mg (1.43mmol) of 4- [3- (dimethylamino) prop-1-ynyl ] phenol in THF (9mL/mmol) gives 0.28g (37%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.34(d,2H),6.91(d,2H),4.26(t,2H),4.03(t,2H),3.78(s,3H),3.40(s,2H),3.25(t,2H),2.88(t,2H),2.31(s,3H),2.22(s,6H),2.08(qn,2H),2.03(qn,2H)；¹³C NMR(125MHz,DMSO-d₆)δppm 163.1,158.9,155.3,151.7,151.3,142.7,136.2,134.9,133.3,129.0,115.2,115.0,85.2,84.1,67.1,52.0,48.3,46.3,44.3,30.8,24.1,23.1,19.7,15.7；HRMS-ESI(m/z):[M+H]⁺C₂₇H₃₁ClN₅O₃Calculated value of S is 540.1836, found value is 540.1834.

Step H2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (dimethylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid methyl ester

Using Buchwald general procedure I, 0.27G of the product of step G (0.5mmol) was used as starting material to give 0.29G (89%) of the desired product.

¹H NMR(500MHz,DMSO-d₆)δppm 7.83(dm,1H),7.50(dm,1H),7.36(m,1H),7.35(m,2H),7.18(m,1H),6.94(m,2H),4.28(m,2H),4.09(t,2H),3.80(s,3H),3.39(s,2H),3.29(t,2H),2.88(t,2H),2.35(s,3H),2.23(s,6H),2.13(m,2H),2.07(m,2H)；HRMS-ESI(m/z):[M+H]⁺C₃₄H₃₆N₇O₃S₂654.2321, found 654.2322.

Step I2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- (dimethylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

To a solution of the product of step H (280mg,0.43mmol) in a 1:1 mixture of THF and water (10mL/mmol) was added 90mg (5eq) LiOHxH₂O and the reaction mixture was stirred at 50 ℃ for 18 hours. After removal of volatiles, purification by reverse phase preparative chromatography (C18, 0.1% TFA in water and MeCN as eluent) gave 132mg (48%) of the desired compound.

HRMS-ESI(m/z):[M+H]⁺C₃₃H₃₄N₇O₃S₂640.2165, found 640.2160.

Example 142:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- (2-morpholinoethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and morpholine as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₄N₉O₄Calculated value of S is 804.4019, found value is 804.4012.

Example 143:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- [2- (1-piperidinyl) ethoxy]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 12 was used as the starting material and piperidine was used as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₄₅H₅₆N₉O₃Calculated value of S is 802.4227, found value is 802.4223.

Example 144:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- (2-piperazin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 12 and piperazine as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₅N₁₀O₃Calculated value of S803.4179, found value 803.4177.

Example 145:3- [1- [ [3- [2- (azepan-1-yl) ethoxy ] ethanol]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation example 12 and azepane as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₆H₅₈N₉O₃Calculated value of S is 816.4383, found value is 816.4379.

Example 146:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- (4-isopropylpiperazin-1-yl) ethoxy]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 12 and 1-isopropylpiperazine as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₇H₆₁N₁₀O₃Calculated value of S is 845.4649, found value is 845.4646.

Example 147:2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-hydroxypropylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

Step A3- [ tert-butyl (dimethyl) silyl group]oxy-N-prop-2-ynyl-prop-1-amine

A mixture of 0.70mL (3.0mmol) 3-bromopropoxy-tert-butyl-dimethyl-silane, 1.9mL (10eq) propargylamine and 1.6mL (3eq) DIPEA in acetonitrile (15mL) was stirred at 50 ℃ until no further conversion was observed. The reaction mixture was concentrated, diluted with DCM and saturated NaHCO₃And brine extraction. The combined organic layers were dried and concentrated to give the desired product in quantitative yield.

¹H NMR(500MHz,dmso-d6)δppm 3.62(t,2H),3.27(d,2H),3.02(t,1H),2.59(t,2H),2.19(brs,1H),1.57(m,2H),0.86(s,9H),0.02(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 73.9,61.5,45.2,37.9,32.7,26.3,-4.8；HRMS(EI)(m/z):[M-CH3]+C₁₁H₂₂Calculated NOSi is 212.1471, found 212.1467.

Step B5- [3- [4- [3- [ tert-butyl (dimethyl) silyl group]Oxopropylamino radical]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl) thiazole-4-carboxylic acid ethyl ester

Using the Sonogashira general procedure starting from 1.0g (1.64mmol) of the product of preparation 15 and 737mg (2eq.) of the product of step A as the appropriate alkyne gives 1.16g (96%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 45.2(t,2H),7.24(dd,1H),7.17(dd,1H),7.14(t,1H),4.27(br.,2H),4.25(q,2H),4.12(t,2H),3.65(t,2H),3.6(s,2H),3.25(t,2H),2.89(t,2H),2.32(s,3H),2.11(m,2H),2.04(m,2H),1.63(m,2H),1.28(t,3H),0.84(s,9H),0.02(s,6H)；

¹³C NMR(500MHz,dmso-d6)δppm 128.8,119.1,115.4,68.3,61.3,60.7,46.3,45.2,38.4,32.4,30.8,26.3,24.2,23.1,19.7,15.7,14.6,-4.8；HRMS-ESI(m/z):[M+H]+C₃₅H₄₈ClFN₅O₄Calculated value of SSi 716.2869, found value 716.2868.

Step C2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [3- [ tert-butyl (dimethyl) silyl group]Oxopropylamino radical ]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid ethyl ester

Using Buchwald general procedure I, starting from 1.16g (1.57mmol) of the product from step B and 730mg (2eq) of 1, 3-benzothiazol-2-amine, 598mg (45%) of the desired product are obtained.

¹H NMR(500MHz,dmso-d6)δppm 7.87(d,1H),7.49(d,1H),7.37(td,1H),7.25(dd,1H),7.19(t,1H),7.17(t,1H),7.17(m,1H),4.26(br.,2H),4.25(q,2H),4.14(t,2H),3.63(t,2H),3.57(s,2H),3.27(t,2H),2.87(t,2H),2.69(t,2H),2.34(s,3H),2.13(m,2H),2.04(m,2H),1.61(m,2H),1.28(t,3H),0.84(s,9H),0.02(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 128.9,126.5,122.5,122.3,119.1,116.3,115.5,68.4,61.3,60.6,46.3,45.2,38.4,32.4,31.1,26.3,23.9,23.2,20.3,14.6,12.9,-4.9；HRMS-ESI(m/z):[M+H]+C₄₂H₅₃FN₇O₄S₂Calculated Si value 830.3354, found 830.3347.

Step D2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [ 2-fluoro-4- [3- (3-hydroxypropylamino) prop-1-ynyl]Phenoxy radical]Propyl radical]Thiazole-4-carboxylic acid

590mg (0.71mmol) of the product of step C and 298mg of LiOHxH₂A mixture of O (10eq) in 7mL THF/water (1:1) was stirred at 60 deg.C until no further conversion was observed. The reaction mixture was treated with 0.71mL (12eq) of concentrated hydrogen chloride (pH 2-3) at 0 ℃ and stirred until no further conversion was observed. After the reaction mixture was concentrated to remove THF and lyophilized, the solid was dissolved in 6N NH₃Is purified by reverse phase chromatography (using 25mM NH)₄HCO₃And MeCN as eluent) to yield 100mg (21%) of the desired product.

HRMS-ESI(m/z):[M+H]+C₃₄H₃₅FN₇O₄S₂688.2176, found 688.2179.

Example 148:2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals ]-5- [3- [4- [3- [ [ (3S) -3, 4-dihydroxybutyl)]-methyl-amino]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

Step A2- [ (4S) -2, 2-dimethyl-1, 3-dioxopentyl-4-yl]Ethyl 4-methylbenzenesulfonate

To a solution of 1.0g (6.8mmol)2- [ (4S) -2, 2-dimethyl-1, 3-dioxolan-4-yl ] ethanol and 3.8mL (4eq) triethylamine in 34mL DCM at 0 deg.C was added 4.5g (2eq) p-toluenesulfonyl 4-methylbenzenesulfonate. The reaction mixture was stirred until no further conversion was observed, concentrated and treated with diisopropyl ether. The precipitated hydrochloride salt was then filtered off, the mother liquor was concentrated and purified by flash chromatography (silica gel using heptane and EtOAc as eluent) to yield 1.6g (81%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 7.79(dm,2H),7.49(dm,2H),4.08(m,2H),4.00(m,1H),3.91/3.44(dd+dd,2H),2.42(s,3H),1.83/1.77(m+m,2H),1.24/1.20(s+s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 132.7,132.7,130.7,128.1,108.6,72.3,68.7,68.4,32.9,27.2/25.9,21.6；HRMS-ESI(m/z):[M+H]+ C14H21O5S calculated 301.1110, found 301.1107.

Step BN- [2- [ (4S) -2, 2-dimethyl-1, 3-dioxopentyl-4-yl]Ethyl radical]Prop-2-yne-1-amines

A mixture of the product of step A (7.6g,25.3mmol), prop-2-yn-1-amine (16mL,10eq) and DIPEA (13.22mL,3eq) in 127mL MeCN was stirred at 50 ℃ for 16 h. After concentration, dissolve in DCM and use concentrated NaHCO₃The solution and brine were extracted and the combined organic layers were dried and concentrated to give 5.0g (107%) of the desired product, which was used without any further purification.

¹H NMR(500MHz,dmso-d6)δppm 4.07(m,1H),3.98/3.43(dd+t,2H),3.28(m,2H),3.05(t,1H),2.62/2.55(m+m,2H),2.23(brs,1H),1.63/1.59(m+m,2H),1.30(s,3H),1.25(s,3H)；¹³C NMR(500MHz,dmso-d6)δppm 108.2,83.4,74.6,74.1,69.2,45.1,37.8,33.6,27.3,26.2；HRMS(EI)(m/z):[M]⁺C₁₀H₁₇NO₂183.1259, found 183.1260.

Step CN- [2- [ (4S) -2, 2-dimethyl-1, 3-dioxopentyl-4-yl]Ethyl radical]-N-methyl-prop-2-yn-1-amine

To a solution of the product of step B (500mg,2.73mmol) in N, N-dimethylformamide (14mL) was added sodium hydride (120mg,1.1eq) portionwise at 0 ℃. After stirring at 0 ℃ for 0.5 h, the mixture was treated with iodomethane (0.17mL,1eq) and stirred at room temperature for 18 h. With saturated NH₄The reaction was quenched with water and the mixture was then treated with Et₂And (4) extracting. The combined organic phases were dried and concentrated to give the desired product (362mg, 67%). GC/MS (C)₁₁H₁₉NO₂)197[M⁺]。

Step D2- (3-chloro-4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazine-8-yl) -5- [3- [4- [3- [2- [ (4S) -2, 2-dimethyl-1, 3-dioxolan-4-yl]Ethyl-methyl-amino]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid ethyl ester

Using the Sonogashira general procedure starting with 0.548g (0.89mmol) of the product of preparation 15 and 350mg (2eq) of the product of step C as the appropriate alkyne gives 510mg (82%) of the desired product.

LC/MS(C₃₄H₄₂ClFN₅O₅S)686[M+H]⁺。

Step E2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [2- [ (4S) -2, 2-dimethyl-1, 3-dioxopentyl-4-yl ]Ethyl-methyl-amino]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid ethyl ester

Using Buchwald general procedure I, starting from 510mg (0.52mmol) of the product from step D and 234mg (3eq) of 1, 3-benzothiazol-2-amine, 200mg (48%) of the desired product are obtained.

¹H NMR(500MHz,dmso-d6)δppm 7.88(dm,1H),7.49(brd,1H),7.37(m,1H),7.3(dd,1H),7.20(dm,1H),7.19(m,1H),7.16(t,1H),4.26(m,2H),4.25(q,2H),4.14(t,2H),4.04(m,1H),3.98/3.45(dd+dd,2H),3.46(s,2H),3.28(m,2H),2.87(t,2H),2.45/2.39(m+m,2H),2.34(s,3H),2.21(s,3H),2.13(m,2H),2.04(m,2H),1.63(m,2H),1.29(t,3H),1.29(s,3H),1.24(s,3H)；HRMS(ESI)(m/z):[M+H]+C₄₁H₄₇FN₇O₅S₂800.3064, found 800.3064.

Step F2- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]]Pyridazin-8-yl radicals]-5- [3- [4- [3- [ [ (3S) -3, 4-dihydroxybutyl)]-methyl-amino]Prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]Thiazole-4-carboxylic acid

200mg (0.25mmol) of the product of step E and 53mg of LiOHxH₂A mixture of O (5eq) in 5mL THF/water (1:1) was stirred at 60 ℃ for 18 h. The reaction mixture was treated with 0.125mL (6eq) of concentrated hydrogen chloride at 0 ℃ (pH 2-3) and stirred at room temperature, then at 60 ℃ for 0.5 h. The reaction mixture was concentrated to remove THF and lyophilized, then the solid was taken upDissolved in 6N NH₃Is purified by reverse phase chromatography (using 5mM NH)₄HCO₃And MeCN as eluent) to yield 47mg (25%) of the desired product.

HRMS(ESI)(m/z):[M+H]+C₃₆H₃₉FN₇O₅S₂732.2438, found 732.2441.

Example 149:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c) ]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ (4-hydroxyphenyl) methylamino ] amino]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 12 and 4- (aminomethyl) phenol as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₇H₅₄N₉O₄Calculated value of S is 840.4019, found value is 840.4016.

Example 150:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ 2-hydroxyethyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 12 and 2- (methylamino) ethanol as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₃H₅₄N₉O₄Calculated value of S is 792.4019, found value is 792.4019.

Example 151:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ 3-methoxypropyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

The desired product was obtained using the general procedure for amine substitution and hydrolysis using preparation 12 and 3-methoxy-N-methyl-propan-1-amine as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₅H₅₈N₉O₄Calculated value of S is 820.4332, found value is 820.4328.

Example 1523- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-hydroxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Step A5- [ tert-butyl (dimethyl) silyl group]Oxy-1, 3-benzothiazol-2-amines

To a mixture of 2-amino-1, 3-benzothiazol-5-ol (750mg,4.51mmol), DMAP (110mg,0.2eq) and imidazole (399mg,1.3eq) in DMF (23mL) was added tert-butyl (chloro) diphenylsilane (816mg,1.2eq) and the reaction mixture was stirred for 18 h. After quenching the reaction with water and extraction with EtOAc, the combined organic phases were dried, concentrated and purified by column chromatography (silica gel, heptane and EtOAc as eluent) to give the desired product (1.07g, 84.5%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.46(d,1H),7.44(s,2H),6.78(d,1H),6.53(dd,1H),0.95(s,9H),0.17(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 168.1,154.5,154.1,124.0,121.5,114.0,109.6,26.1,18.4,-4.0；¹⁵N NMR(100MHz,DMSO-d₆)δppm 237,79。

Step B6- [3- [ [5- [ tert-butyl (dimethyl) silyl ] group]Oxy-1, 3-benzothiazol-2-yl]Amino group]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- (2-hydroxyethoxy) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

Using Buchwald general procedure I at 130 ℃ for 1 hour, 1.0g (1.57mmol) of the product of preparation 12, step C and 883mg (2eq) of the product of step A as starting material gave 1.1g (80%) of the desired product.

¹H NMR(400MHz,DMSO-d₆):δppm 7.95(d,1H),7.7(d,1H),7.65(br,1H),7.38(s,1H),6.95(br,1H),6.71(brd,1H),4.45(t,1H),4.00(t,2H),3.88(s,2H),3.70(s,3H),3.41(q,2H),3.35(t,2H),2.85(t,2H),2.31(s,3H),2.16(s,3H),1.98(qn,2H),1.39(s,2H),1.32/1.25(d+d,4H),1.18/1.12(d+d,4H),1.08/1.00(d+d,4H),0.97(s,9H),0.87(s,6H),0.21(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.9,137.5,122.3,119.1,115.3,62.1,61.5,58.9,52.6,50.1,47.0,46.1,45.4,43.3,30.2,26.1,24.3,21.7,12.6,10.9,-4.0；HRMS-ESI(m/z):[M+H]+C₄₇H₆₃N₈O₅Calculated value of SSi 879.4411, found value 879.4412.

Step C6- [3- [ [5- [ tert-butyl (dimethyl) silyl ] group]Oxy-1, 3-benzothiazol-2-yl]Amino group]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- [2- (p-tolylsulfonyloxy) ethoxy ] ethoxy]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

To a solution of the product of step B (1.1g,1.26mmol) and triethylamine (0.53mL,3eq) in DCM (13mL) was added p-toluenesulfonyl 4-methylbenzenesulfonate (618mg,1.5eq) and the reaction mixture was stirred for 2 hours. Purification by column chromatography (silica gel, DCM and EtOAc as eluent) afforded the desired product (590mg, 45%).

¹H NMR(400MHz,DMSO-d₆):δppm 7.96(d,1H),7.90-6.40(brs,3H),7.70(d,10H),7.70(m,2H),7.46(m,2H),7.38(s,1H),4.07(m,2H),4.00(m,2H),3.85(s,2H),3.69(s,3H),3.49(m,2H),2.85(t,2H),2.40(s,3H),2.31(s,3H),2.15(s,3H),1.98(m,2H),1.33-0.91(m,12H),0.97(s,9H),0.84(s,6H),0.21(s,6H)；HRMS-ESI(m/z):[M+H]+C₅₄H₆₉N₈O₇S₂Calculated Si value 1033.4500, found 1033.4504.

Step D6- [3- [ [5- [ tert-butyl (dimethyl) silyl ] group]Oxy-1, 3-benzothiazol-2-yl]Amino group]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

To a solution of the product of step C (180mg,0.17mmol) in MeCN (1.7mL) and NMP (1.0mL) was added pyrrolidine (0.10mL,7eq) and the reaction mixture was stirred at 60 ℃ for 18 h. Purification by column chromatography (silica gel, DCM and 0.6M NH) ₃MeOH solution as eluent) to give the desired product (144mg, 89%).

¹H NMR(400MHz,DMSO-d₆):δppm 11.24(brs,1H),7.95(d,1H),7.69(d,1H),7.63(d,1H),7.37(s,1H),6.92(br,1H),6.70(dd,1H),4.00(t,2H),3.87(s,2H),3.70(s,3H),3.43(t,2H),2.85(t,2H),2.46(t,2H),2.41(t,4H),2.31(s,3H),2.16(s,3H),1.98(qn,2H),1.63(t,4H),1.38(s,2H),1.30/1.25(d+d,4H),1.19/1.12(d+d,4H),1.08/0.99(d+d,2H),0.97(s,9H),0.86(s,6H),0.21(s,6H)；¹³C NMR(100MHz,DMSO-d₆)δppm 139.8,137.5,122.6,119.0,115.3,59.5,58.9,56.6,54.5,52.6,50.1,47.0,46.0,46.0,43.3,30.2,26.0,24.2,23.6,21.7,12.6,10.9,-4.0；HRMS-ESI(m/z):[M+H]+C₅₁H₇₀N₉O₄Calculated value of SSi 932.5041, found value 932.5014.

Step E3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-hydroxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

To the product of step D (70mg,0.075mmol) in THF (1.2mL) and water (0.30mL) was added LiOH XH₂O (25.2mg,8eq) and the reaction mixture was stirred at 60 ℃ for 1.5 h. Purification by preparative reverse phase HPLC (C18,5mM NH)₄HCO₃(aqueous solution) andIPA as eluent) to give the desired product (45mg, 74%).

HRMS-ESI(m/z):[M+H]+C₄₄H₅₄N₉O₄Calculated value of S is 804,4019, found value is 804.4019.

Example 153:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ 4-hydroxybutyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 12 was used starting with 4- (methylamino) butan-1-ol as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]+C₄₅H₅₈N₉O₄Calculated value of S is 820.4332, found value is 820.4339.

Example 154:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- (dimethylamino) ethoxy ] group]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 14 and dimethylamine as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₀H₄₈N₉O₃Calculated value of S is 734.3601, found value is 734.3589.

Example 155:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [ 5-methyl-1- [ [3- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 14 and pyrrolidine as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₀N₉O₃Calculated value of S is 760.3757, found value is 760.3730.

Example 156:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [ 5-methyl-1- [ [3- [2- (4-methylpiperazin-1-yl) ethoxy ] ethyl]-1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 14 and 1-methylpiperazine as the appropriate amine, the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₃H₅₃N₁₀O₃Calculated value of S789.4017, found value 789.4023.

Example 157:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [ 5-methyl-1- [ [3- (2-morpholinoethoxy) -1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 14 and morpholine as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₀N₉O₄Calculated value of S is 776.3706, found value is 776.3697.

Example 158:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- (3-hydroxypropylamino) ethoxy ] ethyl]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 14 and 3-aminopropan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₁H₅₀N₉O₄Calculated value of S is 764.3706, found value is 764.3700.

Example 159:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- (4-hydroxybutylamino) ethoxy ] ethyl]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 14 and 4-aminobutan-1-ol as the appropriate amine, the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₂N₉O₄Calculated value of S778.3863, found value 778.3859.

Example 160:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ [ (3S) -3, 4-dihydroxybutyl)]Amino group]Ethoxy radical]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 14 and using 2- [ (4S) -2, 2-dimethyl-1, 3-dioxolan-4-yl ] ethanamine as the appropriate amine, the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₂N₉O₅Calculated value of S is 794.3812, found value is 794.3807.

Example 161:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] group]Amino group]Ethoxy radical]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 14 was used starting with 2- (aminomethyl) propane-1, 3-diol as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₂N₉O₅Calculated value of S is 794.3812, found value is 794.3808.

Example 162:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ 4-hydroxybutyl (methyl) amino group ]Ethoxy radical]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 14 was used starting with 4- (methylamino) butan-1-ol as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₄₃H₅₄N₉O₄Calculated value of S is 792.4019, found value is 792.4020.

Example 163:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ 3-hydroxypropyl (methyl) amino group]Ethoxy radical]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 14 and 3- (methylamino) propan-1-ol as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₂N₉O₄Calculated value of S is 778.3863, found value is 778.3858.

Example 164:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ bis (3-hydroxypropyl) amino group]Ethoxy radical]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 14 and using 3- (3-hydroxypropylamino) propan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₆N₉O₅Calculated value of S is 822.4125, found value is 822.4121.

Example 165:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [ 5-methyl-1- [ [3- (2-piperazin-1-ylethoxy) -1-adamantyl]Methyl radical]Pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 14 and piperazine as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₆N₉O₅Calculated value of S is 775.3866, found value is 775.3859.

Example 166:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- (3-pyrrolidin-1-ylpropyl) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 13 and pyrrolidine as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₅H₅₆N₉O₂Calculated value of S is 786.4278, found value is 786.4273.

Example 167:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- (dimethylamino) propyl ] amino]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 13 and dimethylamine as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₃H₅₄N₉O₂Calculated value of S760.4115, found value 760.4121.

Example 168:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- [3- (4-methylpiperazin-1-yl) propyl ] methyl]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 13 and 1-methylpiperazine as the appropriate amine, the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₆H₅₉N₁₀O₂Calculated value of S is 815.4543, found value is 815.4534.

Example 169:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- (3-morpholinopropyl) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 13 and morpholine as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₅H₅₆N₉O₃Calculated value of S is 802.4227, found value is 802.4221.

Example 170:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- (3-hydroxypropylamino) propyl ] amino]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 13 and using 3-aminopropan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₆N₉O₃Calculated value of S is 790.4227, found value is 790.4220.

Example 171:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- (4-hydroxybutylamino) propyl ] amino]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 13 and using 4- (amino) butan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₅H₅₈N₉O₃Calculated value of S is 804.4383, found value is 804.4377.

Example 172:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- [ [ (3S) -3, 4-dihydroxybutyl)]Amino group]Propyl radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 13 and using 2- [ (4S) -2, 2-dimethyl-1, 3-dioxolan-4-yl ] ethanamine as the appropriate amine, the desired product is obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₂N₉O₅Calculated value of S is 820.4332, found value is 820.4328.

Example 173:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] group]Amino group]Propyl radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, preparation 13 was used as the starting material and 2- (aminomethyl) propane-1, 3-diol was used as the appropriate amine to give the desired product.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₂N₉O₅Calculated value of S is 820.4332, found value is 820.4329.

Example 174:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- [ 4-hydroxybutyl (methyl) amino group]Propyl radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 13 and using 4- (methylamino) butan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₃H₅₄N₉O₄Calculated value of S is 818.4540, found value is 818.4536.

Example 175:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- [ 3-hydroxypropyl (methyl) amino group]Propyl radical]-5, 7-dimethyl-1-adamantyl ]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 13 and 3- (methylamino) propan-1-ol as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₂N₉O₄Calculated value of S804.4383, found value 804.4380.

Example 176:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3- [3- [ bis (3-hydroxypropyl) amino group]Propyl radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, starting from preparation 13 and using 3- (3-hydroxypropylamino) propan-1-ol as the appropriate amine, the desired product was obtained.

HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₆N₉O₅Calculated value of S is 848.4645, found value is 848.4645.

Example 177:6- [3- (1, 3-benzothiazol-2-ylamino) -4-methyl-6, 7-dihydro-5H-pyrido [2, 3-c)]Pyridazin-8-yl radicals]-3- [1- [ [3, 5-dimethyl-7- (3-piperazin-1-ylpropyl) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Using the general procedure for amine substitution and hydrolysis, the desired product was obtained using preparation 13 and piperazine as the appropriate amine.

HRMS-ESI(m/z):[M+H]⁺C₄₄H₅₆N₉O₅Calculated value of S is 801.4387, found value is 801.4370.

Example 178:3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Step A:6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- (2-hydroxyethoxy) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid methyl ester

Using Buchwald general procedure I at 130 ℃ for 2 h, using 130mg (0.2mmol) of the product of preparation 12, step C and 52mg (1.5eq) of 7-fluoro-1, 3-benzothiazol-2-amine as starting materials, 139mg (88%) of the desired product were obtained.

¹H NMR(500MHz,dmso-d6)δppm 7.95(d,1H),7.71(d,1H),7.45-7.35(m,1H),7.45-7.35(br.,1H),7.38(s,1H),7.05(m,1H),4.46(br.,1H),4(t,2H),3.88(s,2H),3.71(s,3H),3.41(q,2H),3.35(t,2H),2.87(t,2H),2.33(s,3H),2.16(s,3H),1.99(m,2H),1.39(s,2H),1.30/1.25(d+d,4H),1.18/1.12(d+d,4H),1.07/1.00(d+d,2H),0.87(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 157.1,140.0,137.5,127.7,119.3,108.3,62.1,61.5,59.0,52.7,50.1,47.0,46.0,45.5,43.3,30.2,24.3,21.6,12.5,10.9；HRMS-ESI(m/z):[M+H]⁺C₄₁H₄₈FN₈O₄Calculated value of S is 767.3482, found value is 767.3503.

And B:3- [1- [ [3, 5-dimethyl-7- [2- (p-tolylsulfonyloxy) ethoxy ] ethoxy]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid methyl ester

To a solution of the product of step A (130mg,0.17mmol) and triethylamine (0.071mL,3eq) in DCM (2mL) was added p-toluenesulfonyl 4-methylbenzenesulfonate (83mg,1.5eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, DCM and EtOAc as eluent) gave the desired product (54mg, 34%).

¹H NMR(500MHz,dmso-d6)δppm 7.96(d,1H),7.77(d,2H),7.71(d,1H),7.63-7.26(br.,1H),7.46(d,2H),7.40(br.,1H),7.39(s,1H),7.05(br.,1H),4.06(m,2H),4.00(t,2H),3.85(s,2H),3.69(s,3H),3.49(m,2H),2.87(t,2H),2.41(s,3H),2.33(s,3H),2.15(s,3H),1.99(m,2H),1.28(s,2H),1.20-1.06(m,4H),1.20-1.06(m,4H),1.02/0.97(d+d,2H),0.84(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 140.0,137.6,130.6,128.1,127.6,119.3,108.3,71.5,58.9,58.4,52.6,49.9,46.6,45.9,45.5,43.0,30.1,24.3,21.6,21.6,12.5,10.9；HRMS-ESI(m/z):[M+H]⁺C₄₁H₄₈FN₈O₄Calculated value of S921.3592, found value 921.3567.

Step C:3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

The desired product is obtained by the general procedure of amine substitution and hydrolysis starting from the product of step B and pyrrolidine as the appropriate amine.

HRMS-ESI(m/z):[M+H]+C₄₄H₅₃FN₉O₃Calculated value of S is 806.3976, found value is 806.3974.

Example 179:3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Step A:3- [1- [ [3- (2-hydroxyethoxy) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid methyl ester

Using Buchwald general procedure I at 130 ℃ for 1.5 h, 140mg (0.22mmol) of the product of preparation 12, step C and 54.3mg (1.5eq) of 5-methyl-1, 3-benzothiazol-2-amine were used as starting materials to give 126mg (75%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 12.08/10.89(brs/brs,1H),7.95(d,1H),7.69(d,1H),7.67(br,1H),7.38(s,1H),7.30(br,1H),7.00(d,1H),4.46(brs,1H),4.00(t,2H),3.88(s,2H),3.70(s,3H),3.41(t,2H),3.35(t,2H),2.85(t,2H),2.39(s,3H),2.32(s,3H),2.16(s,3H),1.98(qn,2H),1.39(s,2H),1.30/1.25(d+d,4H),1.18/1.12(d+d,4H),1.08/1.02(d+d,2H),0.87(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 139.8,137.5,123.6,121.6,119.0,62.1,61.5,59.0,52.7,50.1,47.0,46.0,45.4,43.3,30.2,24.3,21.7,21.6,12.6,10.9；HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₁N₈O₄Calculated value of S763.3760, found value 763.3754.

And B, step B:3- [1- [ [3, 5-dimethyl-7- [2- (p-tolylsulfonyloxy) ethoxy ] ethoxy]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid methyl ester

To a solution of the product of step A (119mg,0.16mmol) and triethylamine (0.066mL,3eq) in DCM (2mL) was added p-toluenesulfonyl 4-methylbenzenesulfonate (76mg,1.5eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, DCM and EtOAc as eluent) gave the desired product (93mg, 65%).

¹H NMR(500MHz,dmso-d6)δppm 12.17/10.83(brs/brs,1H),7.95(d,1H),7.77(d,2H),7.7(d,1H),7.69(br,1H),7.46(d,2H),7.42(br,1H),7.39(s,1H),7.00(d,1H),4.07(t,2H),4(t,2H),3.96(s,3H),3.85(s,2H),3.49(t,2H),2.85(t,2H),2.40(s,3H),2.39(s,3H),2.32(s,3H),2.15(s,3H),1.99(qn,2H),1.29(s,2H),1.17/1.1(d+d,4H),1.12/1.1(d+d,4H),1.02/0.97(d+d,2H),0.84(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 139.8,137.6,130.6,128.1,123.6,119.0,71.5,58.8,58.4,52.7,49.9,46.6,45.9,45.4,43.0,30.1,24.3,21.6,21.6,21.6,12.6,10.9；HRMS-ESI(m/z):[M+H]⁺C₄₉H₅₇N₈O₆S₂917.3842, found 917.3840.

And C:3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazole-4-Base of]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₄₅H₅₆N₉O₃Calculated value of S is 802.4227, found value is 802.4220.

Example 180:3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl ]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Step A:3- [1- [ [3- (2-hydroxyethoxy) -5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid methyl ester

Using Buchwald general procedure I at 130 ℃ for 2.5 h, 140mg (0.22mmol) of the product of preparation 12, step C and 60mg (1.5eq) of 5-methyl-1, 3-benzothiazol-2-amine were used as starting materials to give 129mg (75%) of the desired product.

¹H NMR(500MHz,dmso-d6)δppm 7.95(d,1H),7.69(d,1H),7.67(br.,1H),7.38(s,1H),7.02(br.,1H),6.80(dd,1H),4.46(br.,1H),4.00(t,2H),3.88(s,2H),3.80(s,3H),3.70(s,3H),3.41(t,2H),3.35(t,2H),2.85(t,2H),2.32(s,3H),2.16(s,3H),1.98(m,2H),1.39(s,2H),1.30/1.25(d+d,4H),1.18/1.12(d+d,4H),1.08/1(d+d,2H),0.87(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 139.8,137.5,122.6,119.0,110.5,62.1,61.5,58.9,55.8,52.6,50.1,47.0,46.0,45.4,43.3,30.2,24.3,21.7,12.6,10.9；HRMS-ESI(m/z):[M+H]⁺C₄₂H₅₁N₈O₅Calculated value of S is 779.3703, found value is 779.3687.

And B:3- [1- [ [3, 5-dimethyl-7- [2- (p-tolylsulfonyloxy) ethoxy ] ethoxy]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid methyl ester

To a solution of the product of step A (122mg,0.16mmol) and triethylamine (0.066mL,3eq) in DCM (2mL) was added p-tolylsulfonyl 4-methylbenzenesulfonate (77mg,1.5eq) and the reaction mixture was stirred for 1 hour. Purification by column chromatography (silica gel, DCM and EtOAc as eluent) gave the desired product (79mg, 54%).

¹H NMR(500MHz,dmso-d6)δppm 12.17/10.83(brs/brs,1H),7.95(d,1H),7.77(d,2H),7.72(d,1H),7.67(brd,1H),7.46(d,2H),7.39(s,1H),7.02(br,1H),6.80(d,1H),4.07(t,2H),4.00(t,2H),3.86(s,2H),3.80(s,3H),3.69(s,3H),3.49(t,2H),2.86(t,2H),2.41(s,3H),2.33(s,3H),2.15(s,3H),1.99(qn,2H),1.29(s,2H),1.17/1.1(d+d,4H),1.12/1.10(d+d,4H),1.02/0.97(d+d,2H),0.84(s,6H)；¹³C NMR(500MHz,dmso-d6)δppm 139.9,137.6,130.6,128.1,119.0,110.6,71.5,58.8,58.4,55.9,52.6,49.9,46.6,45.9,45.8,43.0,30.1,24.3,21.6,21.6,12.7,10.9；HRMS-ESI(m/z):[M+H]⁺C₄₉H₅₇N₈O₇S₂933.3792, found 933.3794.

Step C:3- [1- [ [3, 5-dimethyl-7- (2-pyrrolidin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

HRMS-ESI(m/z):[M+H]+C₄₅H₅₇N₉O₄Calculated value of S is 818.4176, found value is 818.4172.

The compounds of example 181-219 below were synthesized using the general procedure for amine substitution and hydrolysis starting from a benzothiazole derivative and the appropriate amine as one of preparation examples 12, 13, 14 or similar.

Example 181:3- [1- [ [3, 5-dimethyl-7- [2- (methylamino) ethoxy ] ethanol]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 182:3- [1- [ [3- [2- (dimethylamino) ethoxy ] ethoxy]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c ]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 183:3- [1- [ [3, 5-dimethyl-7- [2- (4-methylpiperazin-1-yl) ethoxy ] ethyl]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 184:6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- (3-hydroxypropylamino) ethoxy ] ethyl]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Example 185:6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl group-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- (4-hydroxybutylamino) ethoxy ] ethyl]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Example 186:3- [1- [ [3- [2- [ [ (3R) -3, 4-dihydroxybutyl)]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 187:3- [1- [ [3- [2- [ [ (3S) -3, 4-dihydroxybutyl)]Amino group ]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 188:3- [1- [ [3, 5-dimethyl-7- (2-piperazin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 189:3- [1- [ [3, 5-dimethyl-7- (2-morpholinoethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 190:6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ 3-hydroxypropyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Example 191:6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ 4-hydroxybutyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl ]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Example 192:6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]-3- [1- [ [3- [2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] group]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]Pyridine-2-carboxylic acid

Example 193:3- [1- [ [3- [2- [ bis (3-hydroxypropyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (7-fluoro-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 194:3- [1- [ [3, 5-dimethyl-7- [2- (methylamino) ethoxy ] ethanol]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 195:3- [1- [ [3- [2- (dimethylamino) ethoxy ] ethoxy]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 196:3- [1- [ [3, 5-dimethyl-7- [2- (4-methylpiperazin-1-yl) ethoxy ] ethyl ]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 197:3- [1- [ [3- [2- (3-hydroxypropylamino) ethoxy ] ethyl]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 198:3- [1- [ [3- [2- (4-hydroxybutylamino) ethoxy ] ethyl]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 199:3- [1- [ [3- [2- [ [ (3R) -3, 4-dihydroxybutyl)]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 200:3- [1- [ [3- [2- [ [ (3S) -3, 4-dihydroxybutyl)]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino ]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 201:3- [1- [ [3, 5-dimethyl-7- (2-piperazin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 202:3- [1- [ [3, 5-dimethyl-7- (2-morpholinoethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl group-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 203:3- [1- [ [3- [2- [ 3-hydroxypropyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 204:3- [1- [ [3- [2- [ 4-hydroxybutyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 205:3- [1- [ [3- [2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] group ]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 206:3- [1- [ [3- [2- [ bis (3-hydroxypropyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-formic acid

Example 207:3- [1- [ [3, 5-dimethyl-7- [2- (methylamino) ethoxy ] ethanol]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 208:3- [1- [ [3- [2- (dimethylamino) ethoxy ] ethoxy]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 209:3- [1- [ [3, 5-dimethyl-7- [2- (4-methylpiperazin-1-yl) ethoxy ] ethyl]-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino ]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 210:3- [1- [ [3- [2- (3-hydroxypropylamino) ethoxy ] ethyl]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 211:3- [1- [ [3- [2- (4-hydroxybutylamino) ethoxy ] ethyl]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 212:3- [1- [ [3- [2- [ [ (3R) -3, 4-dihydroxybutyl)]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 213:3- [1- [ [3- [2- [ [ (3S) -3, 4-dihydroxybutyl)]Amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals ]Pyridine-2-carboxylic acid

Example 214:3- [1- [ [3, 5-dimethyl-7- (2-piperazin-1-ylethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 215:3- [1- [ [3, 5-dimethyl-7- (2-morpholinoethoxy) -1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 216:3- [1- [ [3- [2- [ 3-hydroxypropyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 217:3- [1- [ [3- [2- [ 4-hydroxybutyl (methyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 218:3- [1- [ [3- [2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] group]Amino group]Ethoxy radical ]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 219:3- [1- [ [3- [2- [ bis (3-hydroxypropyl) amino group]Ethoxy radical]-5, 7-dimethyl-1-adamantyl]Methyl radical]-5-methyl-pyrazol-4-yl]-6- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Pyridine-2-carboxylic acid

Example 220:5- [3- [4- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [ 4-methyl-3- [ (5-methyl-1, 3-benzothiazol-2-yl) amino]-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

Example 221:5- [3- [4- [3- (dimethylamino) prop-1-ynyl]-2-fluoro-phenoxy]Propyl radical]-2- [3- [ (5-methoxy-1, 3-benzothiazol-2-yl) amino]-4-methyl-6, 7-dihydro-5H-pyrido [2,3-c]Pyridazin-8-yl radicals]Thiazole-4-carboxylic acid

Pharmacological study

Example A: fluorescence polarization analysis data

Fluorescence polarization measures the rotation of a fluorescent substance in solution, the larger the molecule, the more polarized the fluorescence emission.

Fluorescence-based PUMA from Biopeptides (B)Probe Fluorescein-beta Ala-Ahx-AREIGAQLRRMADDLNAQY-OH of primary accession number Q9BXH1-SEQ ID:01) with a probe having the amino acid sequence (SEQ ID: 02): MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQG WQATFGGGDHPPKSDLIEGRGIPEFEFSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEMETPSAINGNPSWHLADSPAVNGATGHSSSLDAREVIPMAAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVNWGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQER](GST Primary accession numbers P08515 and BCLXLGST (1-218) - (FACTOR _ XA) -hsBCLXL(2-209) 2-209) the bonding results in an increase in anisotropy. If a compound that competitively binds to the same site as the probe to release the probe is added, anisotropy may decrease due to the increase in the amount of free probe.

An 11-point serial dilution of each compound was prepared in DMSO, with final buffer conditions of 10mM 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid [ HEPES]150mM NaCl, 0.05% Tween 20, pH 7.4 and 5% DMSO. The final protein concentration in the assay was 20nM and the fluorescent probe was present at 10 nM. The experiment was incubated at 23 ℃ for 2 hours and then fluorescence polarization (excitation 485nm, emission 525nm, parallel and vertical readings) was measured on a Biotek synergy neo plate reader. Dose response curves were plotted using a 4 parameter Logistic model (sigmoidal dose response model) with XL-Fit software and the Inhibitory Concentration (IC) that increased the fluorescence intensity by 50% (IC)₅₀). According to Cer et al, Nucleic Acids Res,2009, Jul 1; 37(WebServer, Inc.: W441-W445, Slave IC₅₀Value determination K_IThe value is obtained.

The results are summarized in table 1. They show that the compounds of the present invention inhibit the interaction between the Bcl-xL protein and the fluorescent peptide described above.

TABLE 1

Example B: quenching analytical data

Fluorescence quenching analysis measurement of C-terminal Cy 5-labeled BCL-xL protein His-His- (EK) -hsBCLXL(2–197)[N197C](Primary accession number Q07817-1) change in fluorescence intensity upon binding to a C-terminal tag peptide derived from PUMA (UniProtKB primary accession number Q9BXH1), wherein the His-His- (EK) -hsBCLXL(2–197)[N197C]Has an amino acid sequence (SEQ ID:03):

[ MHHHHHHHHGATGSTAGSGTAGSTGASGASTGGTGATHHHHHHHHDDDDKSPMGSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEMETPSAINGNPSWHLADSPAVNGATGHSSSLDAREVIPMAAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVNWGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYG ] which is linked in the C-terminal region to the amino acid X corresponding to the cysteine marked in sulphur with Sulfo-Cyanine5 from Lumiprobe GmbH catalog No. 13380, said PUMA-derived C-terminal marker peptide having the amino acid sequence (SEQ ID:04): [ QWAREIGAQLRRMADDLNAQY ] which is linked in the C-terminal region to the amino acid X ', where X' is the cysteine marked in sulphur with TQ5WS from AAT Bioquest catalog No. 2079.

Addition of a compound that competes for binding to the same site as the peptide will result in an increase in the fluorescence intensity of the protein due to displacement of the fluorescence quencher.

11-Point Serial dilutions of each compound were prepared in DMSO, with final buffer conditions of 10mM 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid [ HEPES]150mM NaCl, 0.05% Tween 20, pH 7.4 and 5% DMSO. The final protein concentration in the assay was 1nM and the peptide was present at 400 nM. The experiment was incubated at 23 ℃ for 2 hours and then the fluorescence intensity was measured on a Biotek SynergyNeo plate reader (excitation 620nm, emission 680 nm). Dose response curves were drawn using a 4 parameter Logistic model (sigmoidal dose response model) with XL-Fit software and measured to increase fluorescence intensityAdding 50% Inhibitory Concentration (IC)₅₀). According to Cer et al, Nucleic Acids Res,2009, Jul 1; 37(WebServer, Inc.: W441-W445, Slave IC₅₀Value determination K_IThe value is obtained.

The results are summarized in table 2.

TABLE 2

The results of tables 1 and 2 show that most compounds of the invention are potent inhibitors of the Bcl-xL protein.

Example C: MTT assay for the Effect of Bcl-xL inhibitors on MOLT-4 or H146 cell viability

The MTT colorimetric method is based on the reduction of tetrazolium salts by living cell mitochondria. Number of viable cells and AProduction of salt is proportional toThe salt can be read spectrophotometrically at 540 nm.

MOLT-4 and H146 cells were purchased from ATCC and cultured in RPMI 1640 supplemented with 10% heat-inactivated fetal bovine serum, penicillin (100IU/ml), streptomycin (100. mu.g/ml) and L-glutamine (2 mM). Cells were incubated at 37 ℃ with 5% CO ₂Is cultured in a humid atmosphere. Cells were seeded in 96-microwell plates (150 μ Ι _ per well) and exposed to compound for 48 hours (3, 16-fold serial dilutions; 9 concentrations each in triplicate). At the end of the incubation period, 15. mu.L of MTT solution (5mg/ml) was added to each well and the cells were incubated for an additional 4 hours. Then, 100. mu.L of 10% Sodium Dodecyl Sulfate (SDS)/HCl 10mM was added to each well, the plate was incubated overnight, and then the optical density was measured at 540 nm.IC₅₀Standard four parameter curve fit calculations were used. IC (integrated circuit)₅₀Defined as the concentration of compound at which the MTT signal decreased to 50% of the control measurement. The results represent the average of at least 2 independent experiments and are given in table 3 below.

TABLE 3

These data indicate that most of these compounds are active in cells and can induce a dose-dependent reduction in the viability of H146 and Molt-4 cell lines.

Example D: pharmacodynamics and tumor regression studies

The in vivo therapeutic and pharmacodynamic effects of Bcl-xL targeted small molecules were determined in MOLT-4 acute T-lymphoblastic leukemia (T-ALL) models administered Intravenously (IV) or orally (PO).

Materials and methods

MOLT-4 cells (ATCC accession number CRL-1582) were cultured in RPMI supplemented with 10% FBS. Cells were resuspended in 50% matrigel (BD Biosciences) and 0.1mL contained 5X 10 ⁶The suspension of individual cells was inoculated subcutaneously into the right flank of female NOD SCID mice (Charles River). For efficacy studies, mice were randomized into groups (7 animals per group) using Easy STAT software when tumors reached the appropriate volume. IV injectionControl vehicle (HPBCD/HCl) or example 24(2.5, 5 or 7.5mg/kg) (twice weekly for 3 weeks-Q3D 6). Mouse body weight was monitored three times a week and tumor size was measured using electronic calipers. Tumor volume was estimated by measuring minimum and maximum tumor diameters using the following formula: (minimum diameter)²(maximum diameter)/2. On the last day (day 31) when at least half of the control animals were still present in the study, tumor growth inhibition was calculated using the formula:

responses were evaluated as follows: if the tumor size is less than or equal to 25mm in at least three consecutive measurements³And is CR (complete response) if the tumor size is measured at least three times consecutively at 25mm³Between half the starting size, PR (partial response). Tumor volume in excess of 2000mm was measured in the first instance³Mice were sacrificed at time or first signs of deterioration of animal health.

For pharmacodynamic studies, when tumors reached the appropriate volume, mice were randomized (3 animals per group) using Easy STAT software. Example 24(7.5mg/kg) was dosed IV in HPBCD/HCl (once daily-QD) or PO in PEG300/EtOH/Phosal (30/10/60) (once daily-QD). Tumor samples were collected 6 hours after dosing and lysed (10mM HEPES pH 7.4, 142.5mM KCl, 5mM MgCl) ₂1mM EDTA, 1% NP40, protease and phosphatase inhibitor cocktail-Calbiochem). Cleared lysates were prepared for immunodetection of cleaved PARP and Caspase 3 by using MSD apoptosis panel whole cell lysis kit (MSD) in 96-well plates according to manufacturer's instructions and analyzed on QuickPlex SQ 120. Whole blood samples were analyzed on a Hematology Analyzer Coulter Ac. T diff (Beckman Coulter).

All experiments were performed according to the current legislation in france after approval by the ethical committee of the Servier institute (IdRS). NOD SCID mice were maintained according to institutional guidelines.

Results

The efficacy of example 24 on MOLT-4 xenografts is shown in FIG. 1. Treatment was started 12 days after tumor cell inoculation (mean size: 235 mm)³). Vehicle (HPBCD/HCl) or example 24(2.5, 5 and 7.5mg/kg) was administered intravenously every 3 days for 6 administrations.

On day 31 after the start of treatment, the tumor growth inhibition (% TGI) induced in example 24 was 77.1% at 2.5mg/kg, 78.7% at 5mg/kg, and 94.7% at 7.5mg/kg (p < 0.05), as shown in FIG. 1 and Table 4. Partial Regression (PR) was achieved in 28.6%, 71.4% and 100% of cases, respectively. Complete Regression (CR) was observed in 14.3% of the highest dose treated animals.

No clinically relevant weight loss due to treatment was observed (fig. 2).

Table 4: MOLT-4 tumor growth inhibition following treatment with example 24(2.5, 5, and 7.5mg/kg, IV dosing, Q3D 6).

Group (A)	Dosage (mg/kg)	％TGI(d31)	％CR	％PR
					Example 24	2.5	77.1*	0	28.6
Example 24	5	78.7*	0	71.4
					Example 24	7.5	94.7*	14.3	100

P value <0.05 compared to control.

Example 24 effects on apoptosis induction and circulating platelet counts of MOLT-4 tumor cells are shown in Table 5. Treatment was started 18 days after tumor cell inoculation (mean size: 461 mm)³). Example 24(7.5mg/kg) was administered once orally or intravenously and samples were collected after 6 hours. This compound showed induction of cleavage of the apoptosis markers, cleaved PARP (24.1-38.6 times untreated control) and cleaved Caspase 3(7.2-18.9 times untreated control), regardless of the route of administration. Furthermore, in view of the well-described role of Bcl-xL in regulating platelet life, it also caused a significant decrease in platelet number (down to 1% of control values).

Table 5: cleaved PARP and cleaved Caspase 3 activation and platelet loss in tumor cells in MOLT-4 transplanted female NOD SCID mice 6 hours after treatment (oral or intravenous) with 7.5mg/kg of example 24.

In summary, we demonstrate herein that the Bcl-xL targeted small molecules described in example 24 are active in vivo after intravenous or oral administration. We observed a strong reduction in tumor regression, apoptosis induction in tumor cells and circulating platelets, consistent with the previously described role of Bcl-xL in apoptosis control and regulation of platelet life (Youle and Straser, Nat Rev. mol Cell biol.2008 Jan; 9 (1): 47-59; Zhang et al, Cell Death Differ.2007 for May.14 (5): 943-51; Mason et al, Cell 2007 Mar 23; 128(6): 1173-86). Furthermore, no clinically relevant weight loss was observed upon treatment with the effective dose, and platelet loss recovered after treatment was discontinued (data not shown). Taken together, these data indicate that there is a possible therapeutic margin using these Bcl-xL targeting small molecules in cancer therapy.

Example E: in vivo pharmacodynamic Properties of Compounds of formula (I)

The pharmacokinetic properties of the compounds of formula (I) are evaluated in rodents (mice, rats) after the PO and/or IV pathway. The formulation is selected based on the physicochemical properties of the drug under test and the route of administration. Single doses of drug prepared as modified formulations (< 5mg/kg) were administered to animals (3 animals per route) via the IV (bolus or 10 minute infusion) or PO (gavage) routes. Blood samples were collected from each animal (up to 6 samples per animal) 24 hours after dosing, and plasma concentrations of test compounds were determined by liquid chromatography combined with tandem mass spectrometry (LC/MS-MS) after extraction.

In some cases, the following protocol was used to determine the pharmacokinetic properties of the compounds of the invention in Wistar rats:

the drug was prepared as a formulation consisting of a mixture of polyethylene glycol 300/absolute ethanol/NaCl 0.9% (40/10/50 v/v/v). The formulation was administered to male Wistar rats (3 animals) by the IV route at a dose of 0.75mg/kg (10 min infusion, 5 mL/kg). Blood samples were taken from each animal at the following time points: end of infusion (10 min), 0.5 hours, 1 hour, 3 hours, 6 hours and 24 hours post-dose. After extraction, plasma concentrations of test compounds were determined by liquid chromatography combined with tandem mass spectrometry detection (LC/MS-MS).

The lower limit of quantitation was 2.5 ng/mL.

The results allow the compounds of the invention to be ranked based on their plasma exposure, elimination rate constant, clearance rate and volume of distribution in order to assess the therapeutic range of the compounds in animal models.

Claims

1. A compound of formula (IA):

wherein:

n is 0, 1 or 2,

◆represents a single bond or a double bond,

◆Z₁represents a bond,-N (R) -or-O-, wherein R represents hydrogen or linear or branched C₁-C₆An alkyl group, a carboxyl group,

◆R₁represents a group selected from: hydrogen; optionally substituted by hydroxy or C ₁-C₆Alkoxy-substituted straight or branched C₁-C₆An alkyl group; c₃-C₆A cycloalkyl group; a trifluoromethyl group; straight or branched C₁-C₆Alkylene-heterocycloalkyl, in which the heterocycloalkyl is optionally substituted by a straight-chain or branched C₁-C₆Alkyl substitution;

◆R₂represents hydrogen or methyl;

◆R_aand R_bIndependently of one another, represent a group selected from: hydrogen; a heterocycloalkyl group; -SO₂Phenyl, wherein the phenyl radical may be linear or branched C₁-C₆Alkyl substitution; straight or branched chain C optionally substituted by one or two hydroxy groups₁-C₆An alkyl group; c₁-C₆alkylene-SO₂OH；C₁-C₆alkylene-SO₂O^-；C₁-C₆alkylene-COOH; c₁-C₆alkylene-PO (OH)₂；C₁-C₆alkylene-NR_dR_e；C₁-C₆alkylene-N⁺R_dR_eR_f；C₁-C₆Alkylene-phenyl, in which the phenyl radical may be substituted by C₁-C₆Alkoxy substitution;

the following groups:

or R_aAnd R_bForm a ring B with the nitrogen atom carrying them₁；

or R_dAnd R_eForm a ring B with the nitrogen atom carrying them₂，

◆Het₁represents a group selected from:

◆Het₂represents a group selected from:

◆A₁is-NH-, -N (C) ₁-C₃Alkyl), O, S or Se, or a salt thereof,

◆A₂is N, CH or C (R)₅)，

G is selected from:

wherein R is_G4Selected from C optionally substituted by 1 to 3 halogen atoms₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₃-C₆A cycloalkyl group,

◆R₅represents a group selected from: optionally substituted by 1 to 3 halogensAtom-substituted C₁-C₆An alkyl group; c₂-C₆An alkenyl group; c₂-C₆An alkynyl group; a halogen or a group-CN,

◆R₆represents a group selected from:

hydrogen;

-C₂-C₆an alkenyl group;

-X₂-O-R₇；

-X₂-NSO₂-R₇；

-C＝C(R₉)-Y₁-O-R₇；

C₃-C₆a cycloalkyl group;

c optionally substituted by hydroxy₃-C₆A heterocycloalkyl group;

C₃-C₆cycloalkylene-Y₂-R₇；

C₃-C₆heterocycloalkylene-Y₂-R₇，

◆R₇Represents a group selected from: straight or branched C ₁-C₆An alkyl group; (C)₃-C₆) cycloalkylene-R₈(ii) a Or:

wherein Cy represents C₃-C₈A cycloalkyl group,

◆R₁₄and R₁₅Independently of one another, represents hydrogen or methyl, or R₁₄And R₁₅Form a cyclohexyl group with the carbon atom bearing them,

◆R_hand R_iIndependently of one another, represents hydrogen or straight-chain or branched C₁-C₆An alkyl group, a carboxyl group,

◆X₁represents a straight or branched chain C₁-C₄Alkylene optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C₁-C₆The substituent of the alkoxy group is replaced by the group,

◆X₂represents a straight chain or a branchC of the chain₁-C₆Alkylene optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C₁-C₆The substituent of the alkoxy group is replaced by the group,

◆R’_aand R'_bIndependently of one another, represent a group selected from: hydrogen; a heterocycloalkyl group; -SO ₂-phenyl, wherein the phenyl group may be linear or branched C₁-C₆Alkyl substitution; optionally substituted by one or two hydroxy groups or C₁-C₆Alkoxy-substituted straight or branched C₁-C₆An alkyl group; c₁-C₆alkylene-SO₂OH；C₁-C₆alkylene-SO₂O^-；C₁-C₆alkylene-COOH; c₁-C₆alkylene-PO (OH)₂；C₁-C₆alkylene-NR'_dR’_e；C₁-C₆alkylene-N⁺R’_dR’_eR’_f；C₁-C₆alkylene-O-C₁-C₆alkylene-OH; c₁-C₆Alkylene-phenyl in which the phenyl group may be substituted by hydroxy or C₁-C₆Alkoxy substitution; the following groups:

or R'_aAnd R'_bForm a ring B with the nitrogen atom carrying them₃，

◆R’_c、R’_d、R’_e、R’_findependently of one another, represents hydrogen or straight-chain or branched C₁-C₆An alkyl group, a carboxyl group,

or R'_dAnd R'_eForm a ring B with the nitrogen atom carrying them₄，

M is 0, 1 or 2,

p is 1, 2, 3 or 4,

◆B₁、B₂、B₃and B₄Independently of one another represent C₃-C₈Heterocycloalkyl, which group may be: (i) is a monocyclic or bicyclic group, wherein bicyclic group comprises a fused, bridged or spiro ring system, (ii) may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen in addition to the nitrogen atom, (iii) is substituted with one or two groups selected from: fluorine, bromine, chlorine, straight or branched C ₁-C₆Alkyl, hydroxy, -NH₂An oxo group or a piperidyl group,

2. A compound according to claim 1, wherein a₄And A₅All represent nitrogen atoms.

3. A compound according to claim 1 or 2, wherein Z is₁represents-NH-or-O-.

4. A compound according to any one of claims 1 to 3, wherein R₃represents-X₁-NR_aR_bPreference is given to the group-C₂H₅-NH-CH₃。

5. A compound according to claim 1 selected from:

6. a compound according to claim 5, which is a compound of formula (IB):

7. a compound according to any one of claims 1 to 6, wherein R₁Represents a hydrogen atom, a methyl group or a cyclopropyl group, preferably a methyl group.

8. A compound according to any one of claims 1 to 7, wherein Het₁Represents:

9. a compound according to any one of claims 1 to 8, wherein Het₂Represents:

10. a compound according to any one of claims 1 to 8, wherein Het₂Represents:

11. a compound according to claim 9, wherein R₆represents-X₂-O-R₇Wherein X is₂Is a propylene group.

12. A compound according to claim 11, wherein R₇Represents the following groups:

13. a compound according to claim 11, wherein R₇Represents the following groups:

14. A compound according to claim 12 or 13, wherein R₈Represents a group selected from: dimethylamino, diethylamino, diisopropylamino, diisobutylamino, methylamino, ethylamino, ethyl (methyl) amino, 4-methyl-piperazin-1-yl, pyrrolidin-1-yl, azetidin-1-yl, 1-piperidinyl, 4-morpholinyl, 4-difluoropiperidin-1-yl, 3-difluoropiperidin-1-yl, 3-hydroxy-1-piperidinyl, (1S,5R) -3-azabicyclo [3.1.0]Hex-3-yl, 4- (1-piperidinyl) -1-piperidinyl, 3-oxo-2, 8-diazaspiro [4.5]Decan-8-yl, (1S,5R) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl, 2- (dimethylamino) ethylamino, 3-piperazin-1-yl, (3R,5S) -3, 5-dimethylpiperazin-1-yl, (but-3-yn-1-yl) amino, (but-3-yn-1-yl) (methyl) amino, (3-azidopropyl) (methyl) amino (3-aminopropyl) amino, (pent-4-yn-1-yl) amino, methyl (pent-4-yn-1-yl) amino, (prop-2-yn-1-yl) amino, (hex-5-yn-1-yl) amino, di-or tri-methyl,3- [ (hex-5-yn-1-yl) (methyl) amino, (4-azidobutyl) (methyl) amino, [2- (2-hydroxyethoxy) ethyl ](methyl) amino and

15. a compound according to claim 12 or 13, wherein R₈Represents a group selected from: bis [ (3S) -3, 4-dihydroxybutyl)]Amino, [ (3S) -3, 4-dihydroxybutyl]Amino, [ (3R) -3, 4-dihydroxybutyl]Amino, acetyl (methyl) amino, 3-hydroxypropylamino.

16. A compound according to claim 11, wherein R₇Represents:

wherein R is₁₁Selected from the group consisting of 3- (dimethylamino) propyl, 3- (methylamino) propyl, aminomethyl, 2- (dimethylamino) ethyl, 4- (dimethylamino) butyl, 2- (methylamino) ethyl, 4- (methylamino) butyl, 3- (azetidin-1-yl) propyl, 3- (4-methylpiperazin-1-yl) propyl, 3-pyrrolidin-1-ylpropyl, 3-morpholinopropyl, 3- (1-piperidinyl) propyl, 3- [ (1R,5S) -6, 6-difluoro-3-azabicyclo [3.1.0]Hex-3-yl and 3- (3-oxo-2, 8-diazaspiro [4.5 ]]Decan-8-yl) propyl.

17. A compound according to claim 11, wherein R₇Represents a group selected from:

18. the compound according to claim 10In which R is₆Represents:

19. a compound according to claim 18, wherein R₇Represents a group selected from:

wherein R is₈Represents a group selected from: hydrogen, 2- (methylamino) ethoxy, 2- (dimethylamino) ethoxy, 2- [ (2-sulfoethyl) amino ]Ethoxy, 2- [ methyl (2-sulfoethyl) amino]Ethoxy, 4-methylpiperazin-1-yl and

20. a compound according to claim 18, wherein R₇Represents a group selected from:

wherein R is₈Represents a group selected from: 2-pyrrolidin-1-ylethoxy, 2- (4-methylpiperazin-1-yl) ethoxy, 2- [ [ (3R) -3, 4-dihydroxybutyl]-methyl-amino]Ethoxy, 2- (4-hydroxybutylamino) ethoxy, 2- [ [ 3-hydroxy-2- (hydroxymethyl) propyl ] ethyl]Amino group]Ethoxy, 2- [ bis (2-hydroxyethyl) amino]Ethoxy, 2- [ [ 2-hydroxy-1- (hydroxymethyl) ethyl]Amino group]Ethoxy, 2- [2- (2-hydroxyethoxy) ethylamino]Ethoxy, 2- [ bis (3-hydroxypropyl) amino]Ethoxy, 2- (3-hydroxypropylamino) ethoxy, 2- [ bis (4-hydroxybutyl) amino]Ethoxy, 2-morpholineSubstituted ethoxy, 2- (1-piperidinyl) ethoxy, 2-piperazin-1-ylethoxy, 2- (azepan-1-yl) ethoxy, 2- (4-isopropylpiperazin-1-yl) ethoxy, 2- [ (4-hydroxyphenyl) methylamino]Ethoxy, 2- [ 2-hydroxyethyl (methyl) amino]Ethoxy, 2- [ 3-methoxypropyl (methyl) amino]Ethoxy, 2- [ 4-hydroxybutyl (methyl) amino]Ethoxy, 3-pyrrolidin-1-ylpropyl, 3- (dimethylamino) propyl, 3- (4-methylpiperazin-1-yl) propyl, 3-morpholinopropyl, 3- (3-hydroxypropylamino) propyl, 3- (4-hydroxybutylamino) propyl, 3- [ [ (3S) -3, 4-dihydroxybutylamino ] propyl ]Amino group]Propyl, 3-hydroxy-2- (hydroxymethyl) propyl]Amino group]Propyl, 3- [ 4-hydroxybutyl (methyl) amino]Propyl, 3- [ 3-hydroxypropyl (methyl) amino]Propyl, 3- [3- [ bis (3-hydroxypropyl) amino]Propyl, 3-piperazin-1-ylpropyl.

21. A compound according to any one of claims 1, 2 and 6, wherein R₃represents-X₁-PO(OH)₂、-X₁-SO₂(OH)、-X₁-NR_aR_b；-X₁-N⁺R_aR_bR_cWherein R is_aOr R_bOr R is_aAnd R_bBoth of them represent a group selected from C₁-C₆alkylene-SO₂OH、C₁-C₆alkylene-SO₂O^-And C₁-C₆alkylene-PO (OH)₂A group of (1).

22. A compound according to any one of claims 1, 2 and 6, wherein R₈represents-NR'_aR’_b；-N⁺R’_aR’_bR’_c；-NH-X’₂-N⁺R’_aR’_bR’_cWherein R'_aAnd R'_bOr both represent C₁-C₆alkylene-SO₂OH and C₁-C₆alkylene-PO (OH)₂A group of (1).

23. A compound according to claim 1 selected from:

24. A compound according to claim 1 selected from:

25. A pharmaceutical composition comprising a compound according to any one of claims 1 to 24, or an addition salt thereof with a pharmaceutically acceptable acid or base, and one or more pharmaceutically acceptable excipients.

26. The pharmaceutical composition of claim 25 for use as a pro-apoptotic agent.

27. The pharmaceutical composition of claim 25, for use in the treatment of cancer, autoimmune diseases or immune system diseases.

28. The pharmaceutical composition of claim 27, wherein the cancer is a hematologic malignancy or a solid tumor.

29. The pharmaceutical composition according to claim 28, wherein the hematological malignancy is myeloma, in particular multiple myeloma, lymphoma, in particular non-hodgkin's lymphoma (NHL), or leukemia, in particular Chronic Lymphocytic Leukemia (CLL), acute T-lymphocytic leukemia (T-ALL), acute B-lymphocytic leukemia (B-ALL), and Acute Myeloid Leukemia (AML).

30. The pharmaceutical composition of claim 28, wherein said solid tumor is selected from the group consisting of bladder, brain, breast, uterine, esophageal, and liver cancers, colorectal, renal, melanoma, ovarian, prostate, pancreatic, and lung cancers.

31. A compound according to any one of claims 1 to 24, or an addition salt thereof with a pharmaceutically acceptable acid or base, for use in the treatment of a cancer selected from: myeloma, in particular multiple myeloma, lymphoma, in particular non-hodgkin's lymphoma (NHL), or leukemia, in particular Chronic Lymphocytic Leukemia (CLL), acute T-lymphocytic leukemia (T-ALL), acute B-lymphocytic leukemia (B-ALL) and Acute Myeloid Leukemia (AML), bladder cancer, brain cancer, breast cancer, uterine cancer, esophageal cancer and liver cancer, colorectal cancer, kidney cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer and lung cancer, in particular non-small cell lung cancer and small cell lung cancer.

32. A combination of a compound according to any one of claims 1 to 24 and an anticancer agent selected from genotoxic agents, mitotic toxic agents, antimetabolites, proteasome inhibitors, kinase inhibitors and antibodies.

33. A pharmaceutical composition comprising the combination of claim 32 and one or more pharmaceutically acceptable excipients.

34. The combination according to claim 32 for use in the treatment of cancer.

35. A compound according to any one of claims 1 to 24 for use in the treatment of cancer in need of radiotherapy.

36. The pharmaceutical composition according to claim 25, for use in the treatment of a disease or condition characterized by excessive or deregulated platelet activity, in particular a pro-thrombotic condition.

37. A synthetic intermediate selected from the group consisting of:

wherein R is₇As defined in claim 1 and G1 represents C₁-C₆Alkyl, preferably methyl, or (4-methoxyphenyl) methyl.

38. A synthetic intermediate selected from the group consisting of:

wherein R is₆As defined in claim 1 and G1 represents C₁-C₆Alkyl, preferably methyl, or (4-methoxyphenyl) methyl.