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WO2018187294A1 - Combinaisons de composés de pyrimido-pyridazinone, procédés, kits et formulations associées - Google Patents

Combinaisons de composés de pyrimido-pyridazinone, procédés, kits et formulations associées Download PDF

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Publication number
WO2018187294A1
WO2018187294A1 PCT/US2018/025841 US2018025841W WO2018187294A1 WO 2018187294 A1 WO2018187294 A1 WO 2018187294A1 US 2018025841 W US2018025841 W US 2018025841W WO 2018187294 A1 WO2018187294 A1 WO 2018187294A1
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Prior art keywords
pyridazin
amino
phenyl
oxo
piperidin
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PCT/US2018/025841
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English (en)
Inventor
Sanjeeva Reddy
Niranjan Rao
Louis Denis
Sandeep Gupta
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Asana Biosciences, Llc
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Publication of WO2018187294A1 publication Critical patent/WO2018187294A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • Protein kinases constitute a large family of structurally related enzymes that are responsible for the control of a variety of signal transduction processes within cells. Almost all kinases contain a similar 250 to 300 amino acid catalytic domain. The kinases can be categorized into families by the substrates they phosphorylate.
  • JAK (Janus kinase) is a family of intracellular non-receptor tyrosine kinases, which includes JAK1, JAK2, JAK3 and TYK2. JAK is expressed in hematopoietic cells and abundantly in primary leukemic cells from children with acute lymphoblastic leukemia.
  • the downstream substrates of JAK include the signal tranducer activator of transcription (STAT) proteins. STAT proteins function both as signaling molecules and transcription factors and ultimately bind to specific DNA sequences present in the promoters of cytokine-responsive genes.
  • JAK/STAT signaling has been implicated in the mediation of many abnormal immune responses such as allergies, asthma, autoimmune diseases such as transplant (allograft) rejection, rheumatoid arthritis, amyotrophic lateral sclerosis and multiple sclerosis, as well as in solid and hematologic malignancies such as leukemia and lymphomas.
  • Spleen tyrosine kinase is a member of the syk family of protein tyrosine kinases and plays a crucial role in inflammatory and allergic responses. Syk triggers IgE and IgG receptor mediated signaling in mast cells, basophils, and macrophages leading to
  • ITAM-mediated signaling has emerged as a primary event in signaling pathways responsible for human pathologies.
  • ITAM-mediated signaling is responsible for relaying activation signals initiated at classical immune receptors such as T-cell receptors, B-cell receptors, and Fc receptors in immune cells and at GPVI and FcyRIIa in platelets to downstream intracellular molecules such as Syk.
  • the binding of a ligand to an ITAM-containing receptor triggers signaling events which allows for the recruitment of proteins from a family of nonreceptor tyrosine kinases called the Src family. These kinases phosphorylate tyrosine residues within the ITAM sequence, a region with which the tandem SH2 domains on either Syk or ZAP-70 interact.
  • Syk with diphosphorylated ITAM sequences induces a conformation change in the kinases that allows for tyrosine phosphorylation of the kinase itself.
  • these kinases contribute to normal host defense, they also play roles in the pathogenesis of diseases. Many diseases are associated with abnormal cellular responses triggered by protein kinase-mediated events. These diseases include autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, neurological and
  • Immunotherapy has been used as a treatment for various diseases by inducing, enhancing or modulating an immune response.
  • cancer immunotherapy attempts to stimulate the immune system to destroy tumors, or inhibit immune system checkpoint molecules so that they no longer block proteins on cancer cells or immune cells (such as T cells) that respond to them.
  • immune system checkpoint molecules are designed to overcome the ability of cancer cells to mask themselves from the patient's immune system.
  • oncologic therapies e.g., which act by inhibition of specific biochemical pathways
  • immunotherapeutic agents for example immune checkpoint inhibitors or therapies with immunostimulatory potential
  • combination treatments may result in increases in toxicity and adverse side effects, for example by increasing cytokine production and other inflammatory reactions related to the non-tumor suppressive activities of the immunotherapeutic agents.
  • the present disclosure provides pharmaceutical combinations of a compound of Formula (I), wherein R 1 and R 2 are defined herein
  • methods of treating cancer include administering a compound of Formula (I) and at least one immunotherapeutic agent to a patient in need thereof.
  • methods of inhibiting tumor growth or metastasis include administering a compound of Formula (I) and at least one immunotherapeutic agent to a patient in need thereof.
  • dosing regimens include administering to a patient in need thereof a compound of Formula (I) and at least one immunotherapeutic agent to a patient in need thereof.
  • kits include a dosage form comprising a compound of Formula (I) and a dosage form comprising at least one immunotherapeutic agent and therapeutic instructions for administering the dosage forms.
  • uses of a medicament in treating cancer include administering to a patient in need thereof a multi-part pharmaceutical combination including a compound of Formula (I) and at least one immunotherapeutic agent to a patient in need thereof.
  • Figs. 1 and 2 provide comparative data illustrating the anti-inflammatory effects of methotrexate, a known anti-inflammatory, and a compound described herein which is encompassed by the compound of Formula (I), using the Collagen Induced Arthritis (CIA) model of human rheumatoid arthritis (RA) in female lewis rats.
  • CIA Collagen Induced Arthritis
  • RA rheumatoid arthritis
  • Figure 1 A illustrates anti- inflammatory effects as a function of the amount of edema (mL) vs. time (days).
  • the black diamonds ( ⁇ ) represent results for the control.
  • Figure 1 B illustrates anti-inflammatory effects as a function of the amount of edema (mL) vs. time (days).
  • the circles ( ⁇ ) represent results for the control.
  • the inverted triangles represent results for the compound of Example 62.
  • the astericks (*) represent results for the compound of Example 108.
  • the squares ( ⁇ ) represent results for the compound of Example 189.
  • the diamonds ( ⁇ ) represent results for the compound of Example 191.
  • Figure 2A illustrates anti-inflammatory effects as a function of arhtritic score (per rat) vs. time (days).
  • the black diamonds ( ⁇ ) represent results for the control.
  • the triangles represent results for the compound of Example 19.
  • the crosses (x) represent results for methotrexate.
  • Figure 2B illustrates anti-inflammatory effects as a function of arhtritic score (per rat) vs. time (days).
  • the circles ( ⁇ ) represent results for the control.
  • the triangles represent
  • Figure 3 illustrates percent decrease in inflammation biomarkers on Day 15 after initiation of Compoud A treatment with the compound of Example 189 ("COMPOUND A") (BID dosing).
  • the percent change shown in Figure 3 are average values of all patients in each cohort.
  • B2M p2-Microglobulin
  • CRP C-Reactive Protein
  • IL-18 Interleukin-18
  • ⁇ -1 ⁇ Macrophage Inflammatory Protein- 1 ⁇
  • TNFR2 Tumor necrosis factor receptor 2.
  • Figure 4 illustrates average tumor volume (mm 3 ) in mice treated with COMPOUND A alone or in combination with mouse anti-PD 1 antibody after different days of treatment initiation. Table shown in Figure 4 shows % TGI for various treatments on days 5 to 15 and statistical comparisons between groups on day 15.
  • Figure 5 illustrates average tumor volume (mm 3 ) in mice treated with COMPOUND A alone or in combination with epacadostat (IDO-1 inhibitor) after different days of treatment initiation. Table shown in Figure 5 shows % TGI for various treatments on days 5 to 15 and statistical comparisons between groups on day 15.
  • FIG. 6 illustrates changes in average body weight of mice treated with
  • COMPOUND A alone or in combination with mouse anti-PD 1 antibody after treatment on day 15.
  • FIG. 7 illustrates changes in average body weight of mice treated with
  • COMPOUND A alone or in combination with epacadostat after treatment on day 15.
  • Figure 8 illustrates percent change in body weight of mice treated with COMPOUND A alone or in combination with mouse anti-PD 1 antibody measured on different days.
  • Figure 9 illustrates percent change in body weight of mice treated with COMPOUND A alone or in combination with epacadostat measured on different days.
  • Figure 10 presents raw data for tumor volume and body weight measurement on day
  • Figure 11 presents raw data for tumor volume and body weight measurement on day
  • Figure 12 presents raw data for tumor volume and body weight measurement on day
  • Figure 13 presents raw data for tumor volume and body weight measurement on day 11.
  • Figure 14 presents raw data for tumor volume and body weight measurement on day
  • Figure 15 presents raw data for COMPOUND A concentrations in plasma analyzed 2 h after last dose on day 15.
  • Figure 16 presents raw data for COMPOUND A concentrations in tumor analyzed 2 h after last dose on day 15.
  • Figure 17 illustrates average tumor volume (mm 3 ) in mice treated with COMPOUND
  • Table shown in Figure 17 shows % TGI for various treatments on days 4 to 15 and statistical comparisons between groups on day 15.
  • Figure 18 illustrates average tumor volume (mm 3 ) in mice treated with COMPOUND
  • Table shown in Figure 18 shows % TGI for various treatments on days 4 to 15 and statistical comparisons between groups on day 15.
  • FIG 19 illustrates changes in average body weight of mice treated with
  • COMPOUND A alone or in combination with mouse anti-PDl antibody after treatment on day 15.
  • FIG. 20 illustrates changes in average body weight of mice treated with
  • COMPOUND A alone or in combination with epacadostat after treatment on day 15.
  • Figure 21 illustrates percent change in body weight of mice treated with
  • COMPOUND A alone or in combination with epacadostat measured on different days.
  • Figure 23 presents raw data for tumor volume and body weight measurement on day
  • Figure 24 presents raw data for tumor volume and body weight measurement on day
  • Figure 25 presents raw data for tumor volume and body weight measurement on day
  • Figure 26 presents raw data for tumor volume and body weight measurement on day 11. The shaded rows in Figure 26 indicate animal mortality.
  • Figure 27 presents raw data for tumor volume and body weight measurement on day 15. The shaded rows in Figure 27 indicate animal mortality.
  • Figure 28 presents raw data for COMPOUND A concentrations in plasma on day 15 two hours of last dose.
  • Figure 29 presents raw data for COMPOUND A concentration in tumor on day 15 two hours of last dose.
  • Figure 30 illustrates average tumor volume (mm 3 ) in mice treated with COMPOUND A alone or in combination with mouse anti-PD 1 antibody after different days of treatment initiation. Table shown in Figure 30 shows % TGI for various treatments on days 4 to 15 and statistical comparisons between groups on day 15 .
  • Figure 31 illustrates average tumor volume (mm 3 ) in mice treated with COMPOUND A alone or in combination with epacadostat after different days of treatment initiation. Table shown in Figure 31 shows % TGI for various treatments on days 4 to 15 and statistical comparisons between groups on day 15.
  • Figure 32 illustrates changes in average body weight of mice treated with
  • COMPOUND A alone or in combination with mouse anti-PDl antibody after treatment on day 15.
  • Figure 33 illustrates changes in average body weight of mice treated with
  • COMPOUND A alone or in combination with epacadostat after treatment on day 15.
  • Figure 34 illustrates percent change in body weight of mice treated with
  • COMPOUND A alone or in combination with epacadostat measured on different days.
  • Figure 36 presents raw data for tumor volume and body weight measurement on day
  • Figure 37 presents raw data for tumor volume and body weight measurement on day
  • Figure 38 presents raw data for tumor volume and body weight measurement on day
  • Figure 39 presents raw data for tumor volume and body weight measurement on day
  • Figure 40 presents raw data for tumor volume and body weight measurement on day
  • Figure 41 presents raw data for COMPOUND A concentrations in plasma analyzed 2 h after last dose on day 15.
  • Figure 42 presents raw data for COMPOUND A concentrations in tumor analyzed 2 h after last dose on day 15.
  • the disclosure provides pharmaceutical combinations comprising compounds and immunotherapeutic agents, which are capable of reducing or eliminating inflammation caused by tissue insult, injury, or pathology, and treating cancer and other diseases.
  • the compounds disclosed herein can function through a protein kinase inhibitory mechanism.
  • the present disclosure provides a pharmaceutical combination comprising a compound of Formula (I):
  • R 1 is NR 4 R 5 , optionally substituted C 1 to C 6 alkoxy, optionally substituted C 6 to C 14 aryl, optionally substituted heteroaryl, optionally substituted 3-10 membered monocyclic orbicyclic cycloalkyl, or optionally substituted 3-10 membered monocyclic or bicyclic heterocyclyl.
  • the 3-4 membered cycloalkyl and heterocyclyl are saturated.
  • the hydrogen atoms on the same carbon atom of the cycloalkyl or heterocyclyl are optionally replaced with an optionally substituted 3-6 membered cycloalkyl or heterocyclyl to form a spirocycloalkyl or spiroheterocyclyl.
  • the hydrogen atoms on the same atom of the cycloalkyl or heterocyclyl are optionally replaced with O to form an oxo substituent.
  • R 1 is N(C 1 to C 6 alkyl)(C 1 to C 6 alkyl) or C 1 to C 6 alkoxy. ii. In still a further embodiment, R 1 is N(CH(CH 3 ) 2 )2, N(CH 3 ) 2 , OCH 2 CH 3 , or OCH 3 . iii. In another embodiment, R 1 is optionally substituted phenyl.
  • R 1 is of the structure:
  • R 22 , R 23 , R 24 , R 25 , and R 26 are, independently, H, C(O)(C 1 to C 6 alkoxy), C(O)OH, 0(C 1 to C 3 perfluoroalkyl), 0(C 1 to C 6 perfluoroalkoxy), Ci to Ce alkoxy, halogen, (C 1 to C 6 alkyl)heterocyclyl, or (C 1 to C 6 alkyl)CN.
  • R 1 is:
  • R 1 is optionally substituted 5-9 membered saturated heterocyclyl.
  • R 1 is of the structure:
  • R 34 , R 35 , R 36 , and R 37 are, independently, H, C 1 to C 6 alkyl, or CN;
  • R 7 and R 8 are, independently, H, C 1 to C 6 alkyl, C(O)OH, (C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)C(O)OH, C(O)(C 1 to C 6 alkyl)CN, or CN; and
  • x is 0 to 2.
  • R is:
  • R 1 is of the structure:
  • R 1 is:
  • R 1 is a heteroaryl
  • R 1 is thiophene, benzooxole, or pyridine.
  • R 1 is a monocyclic C 3 to C 8 cycloalkyl.
  • R 1 is cycloheptyl or cyclohexyl, both optionally substituted with -N(C 1 to C 6 alkyl)(C 1 to C 6 alkyl).
  • R 1 is piperidine substituted with C(O)(C 1 to C 6 alkyl)CN.
  • xvi. In still a further embodiment, R is:
  • R 2 is phenyl substituted with C(O)NR 4 R 5 .
  • R 2 is phenyl substituted with
  • R 2 is phenyl substituted with NR 4 R 5 .
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 .
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6 alkyl)NR 4 R 5 and R 4 and R 5 are taken together with the nitrogen atom to which they are attached to form a 6-membered ring.
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6
  • R 10 , R 11 , R 12 , and R 13 are, independently, H or C 1 to C 6 alkyl;
  • R 14 is halogen, OH, C(O)OH, C 1 to C 6 alkoxy, (C 1 to C 6 alkyl)halogen, (C 1 to C 6 alkyl)C(O)OH, C 1 to C 6 hydroxyalkyl, C 3 to C 8 cycloalkyl, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O)NH(C 1 to C 6 hydroxyalkyl), (C 1 to C 6 alkyl)C(O)N(C 1 to C 6 hydroxyalkyl) 2 , (C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)heteroaryl, or heteroaryl; and R 18 is C 1 to C 6 hydroxyalkyl or (C 1 to C 6 alkyl)C(O)OH.
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6 alkyl)NR 4 R 5 and R 4 and R 5 are joined to form:
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6
  • R 10 , R 11 , R 12 , and R 13 are, independently, H or C 1 to C 6 alkyl; Y is O or NR 9 ; and R 9 is H, C 1 to C 6 alkyl, OH, C(O)OH, C 1 to C 6 hydroxyalkyl, (Ci to C 6 alkyl)NH 2 , (C 1 to C 6 alkyl)N(C 1 to C 6 alkyl)(C 1 to C 6 alkyl), (C 1 to C 6 alkyl)(C 1 to C 6 alkoxy), C(O)(C 1 to C 6 alkyl)NH 2 , (C 1 to C 6 alkyl)C(O)OH, C(O)(C 1 to C 6 hydroxyalkyl), C(O)(C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)halogen, or (C 1 to C 6 alkyl)0(C 1 to C 6 alkyl;
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6
  • R 2 is phenyl substituted with NR 4 R 5 or (C 1 to C 6
  • alk 4 R 5 and R 4 and R 5 are taken together to form:
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 .
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 and
  • R 4 and R 5 are (C 1 to C 6 hydroxyalkyl).
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 and NR 4 R 5 is:
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 and R 4 and R 5 are joined to form an optionally substituted 6-membered ring.
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 and NR 4 R 5 are joined to form the 6-membered ring:
  • R 14 is H, OH, C(O)OH, C 1 to C 6 alkyl, or (C 1 to C 6 alkyl)CN.
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 and NR 4 R 5 are joined to form the 6-membered ring: wherein, Y is O or NR 9 ; and R 9 is H, C 1 to C 6 alkyl, OH, C 1 to C 6 hydroxyalkyl, C(O)(C 1 to C 6 hydroxyalkyl), C(O)(C 1 to C 6 alkyl)CN, (Ci to C 6 alkyl)CN, (C 1 to C 6 alkyl)NH 2 , (C 1 to C 6 alkyl)halogen, C(O)(C 1 to C 6 alkyl)CN or (C 1 to C 6 alkyl)0(C 1 to C 6 alkyl)C(O)(C 1 to C 6 alkyl)NH 2 .
  • R 2 is phenyl substituted with (C 1 to C 6 alkyl)NR 4 R 5 and
  • NR 4 R 5 are joined to form the 6-membered ring
  • a, b, c, d, and e are, independently, absent, (CH 2 ), CH(R 3 ), or O; and R 3 is H or C(O)OH.
  • R 2 is a heteroaryl substituted with (C 1 to C 6 alkyl)NR 4 R 5 . v. In still another embodiment, R 2 is:
  • R 2 is a heteroaryl substituted with NR 4 R 5 .
  • R 2 is a heteroaryl substituted with NR 4 R 5 and the heteroaryl is pyridine.
  • R 2 is of the structure:
  • R 2 is of the structure:
  • R 80 is OH, -(C 1 to C 6 alkyl)CN, C 1 to C 6 hydroxyalkyl, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)heterocycle or -(C 1 to C 6 alkyl) C(O)OH.
  • R 80 is OH, -(C 1 to C 6 alkyl)CN, C 1 to C 6 hydroxyalkyl, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)heterocycle or -(C 1 to C 6 alkyl) C(O)OH.
  • p is 1 to 6; and R is H or C(O)OH.
  • R is of the structure:
  • R 90 is H, C 1 to C 6 alkyl, C(O)(C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)C(O)OH, or C(O)C 1 to C 6 hydroxyalkyl.
  • heterocyclyl optionally substituted heteroaryl, -O-(C 1 to C 6 alkyl)C(O)OH, -O-(Ci to C 6 alkyl)NR 4 R 5 , -0(optionally substituted heterocycle), -0(C 1 to C 6 alkyl)N(C 1 to C 6 alkyl)(C 1 to C 6 alkyl), -0(C 1 to C 6 alkyl)NH 2 , C 1 to C 6 hydroxyalkyl, -0(C 1 to C 6 hydroxyalkyl), -0(C 1 to C 6 alkyl)C(O)OH, -C 1 to C 6 alkoxy-C 1 to C 6 alkoxy, - 0(heterocycle)(C 1 to C 6 hydroxyalkyl), -S0 2 (C 1 to C 6 alkyl), or -(C 1 to C 6 alkyl)(Ci to Ce alkoxy)halogen.
  • R 2 is of the structure:
  • R 6 is H, (C 1 to C 6 alkyl)C(O)OH, or (C 1 to C 6 alkyl)CN. ee.
  • R 2 is of the structure:
  • z is 1 , 2, 3, 4, 5, or 6.
  • R 2 is of the structure:
  • R 6 is H or (C 1 to C 6 alkyl)C(O)OH.
  • R 2 is -0(C 1 to C 6 alkyl)NR 4 R 5 .
  • R 2 is of the structure:
  • y is 2 to 6; and R is H, OH, C 1 to C 6 alkyl, C 1 to C 6 hydroxyalkyl, or -(C 1 to C 6 alkyl)C(O)OH.
  • R 2 is of the structure:
  • R 2 is of the structure: wherein, r is 2 to 6; and R is H, C(O)OH or C 1 to C 6 hydroxyalkyl.
  • R 2 is:
  • R 2 is aryl substituted with -O-(C 1 to C 6 alkyl)-heterocycle. mm. In a further embodiment, R 2 is:
  • pharmaceutical combination comprising a compound of Formula (I) is provided, wherein R 1 is NR 4 R 5 , optionally substituted C 1 to C 6 alkoxy, optionally substituted C 6 to C 14 aryl, optionally substituted heteroaryl, optionally substituted 3-10 membered monocyclic or bicyclic cycloalkyl, or optionally substituted 3-10 membered monocyclic or bicyclic heterocyclyl.
  • R 1 is NR 4 R 5 , optionally substituted C 1 to C 6 alkoxy, optionally substituted C 6 to C 14 aryl, optionally substituted heteroaryl, optionally substituted 3-10 membered monocyclic or bicyclic cycloalkyl, or optionally substituted 3-10 membered monocyclic or bicyclic heterocyclyl.
  • R 1 is NR 4 R 5 , optionally substituted C 1 to C 6 alkoxy, optionally substituted C 6 to C 14 aryl, optionally substituted heteroaryl, optionally substituted 3-10 membered monocycl
  • Hydrogen atoms on the same carbon atom of the cycloalkyl or heterocyclyl are optionally replaced with an optionally substituted 3-6 membered cycloalkyl or heterocyclyl to form a spirocycloalkyl or spiroheterocyclyl.
  • hydrogen atoms on the same atom of the cycloalkyl or heterocyclyl are optionally replaced with O to form an oxo substituent.
  • R 2 is phenyl or 5-6 membered heteroaryl containing at least one N or NH in the backbone, wherein R 2 is optionally substituted with one or more R 19 and when R 2 is 4- pyridyl, the 4-pyridyl lacks a carbonyl substituent at the 2 nd position.
  • R 19 is NR 4 R 5 , (C 1 to C 6 alkyl)NR 4 R 5 , C 1 to C 6 alkyl, C(O)NR 4 R 5 , C 3 to C 8 cycloalkyl substituted with one or more R 21 , or heterocyclyl substituted with one or more R 21 .
  • R 21 is (C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)C(O)OH, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O)NHCH 2 CH 2 OH, or (C 1 to C 6 alkyl)C(O)N(CH 2 CH 2 OH) 2 .
  • R 4 and R 5 are independently selected from among H, C 1 to C 6 alkyl, C 1 to C 6 hydroxyalkyl, and (C 1 to C 6 alkyl)N(C 1 to C 6 alkyl)(C 1 to C 6 alkyl).
  • R 4 and R 5 are joined to form an optionally substituted 3-8 membered heterocyclyl optionally further containing one or more O, S(O) n , or NR 9 .
  • R 9 is H, OH, (Ci to C 6 alkyl)C(O)OH, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O)NHCH 2 CH 2 OH, (C 1 to C 6 alkyl)C(O)N(CH 2 CH 2 OH) 2 , C(O)(C 1 to C 6 alkyl)NH 2 , C(O)(C 1 to C 6 alkyl)OH, C 1 to C 6 hydroxyalkyl, or C 1 to C 6 alkyl and n is 0 to 2.
  • R 9 is CH 2 CH 2 OH.
  • Hydrogen atoms on the same carbon atom of the heterocyclyl are optionally replaced with a 3-6 membered cycloalkyl or heterocyclyl optionally substituted with one or more R to form a spirocycloalkyl or spiroheterocyclyl.
  • R 20 is C(O)0(C 1 to C 6 alkyl), C(O)OH, (C 1 to C 6 alkyl)C(O)OH, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O)NHCH 2 CH 2 OH, or (C 1 to C 6 alkyl)C(O)N(CH 2 CH 2 OH) 2 .
  • hydrogen atoms on the same atom of any of the heterocyclyls or cycloalkyls of R 9 are optionally replaced with O to form an oxo substituent; or a pharmaceutically acceptable salt or ester thereof.
  • R 1 is NR 4 R 5 , C 1 to C 6 alkoxy, optionally substituted phenyl, heteroaryl, optionally substituted 3-10 membered cycloalkyl, or optionally substituted 3-10 membered monocyclic or bicyclic heterocyclyl.
  • Hydrogen atoms on the same carbon atom of the cycloalkyl or heterocyclyl are optionally replaced with an optionally substituted 3-6 membered cycloalkyl or heterocyclyl to form a spirocycloalkyl or spiroheterocyclyl.
  • R 2 is phenyl or pyrazole, wherein R 2 is optionally substituted with one or more R 19 .
  • R 19 is NR 4 R 5 , (C 1 to C 6 alkyl) R 4 R 5 , Q to Ce alkyl, C(O)NR 4 R 5 , C 3 to C 8 cycloalkyl substituted with one or more R 21 , or heterocyclyl substituted with one or more R 21 .
  • R 21 is (C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)C(O)OH, (Ci to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O) HCH 2 CH 2 OH, (C 1 to C 6
  • R 4 and R 5 are (a) independently selected from among H, Ci to C 6 alkyl, C 1 to C 6 hydroxyalkyl, and (C 1 to C 6 alkyl)N(C 1 to C 6 alkyl)(C 1 to C 6 alkyl) or (b) joined to form an optionally substituted 3-8 membered heterocyclyl optionally further containing one or more O, S(O) impart, or NR 9 .
  • R 9 is H, OH, C 1 to C 6 hydroxyalkyl, (C 1 to C 6 alkyl)C(O)OH, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O)NHCH 2 CH 2 OH, (C 1 to C 6 alkyl)C(O)N(CH 2 CH 2 OH) 2 , C(O)(C 1 to C 6 alkyl)NH 2 , C(O)(C 1 to C 6 alkyl)OH, or C 1 to C 6 alkyl and n is 0 to 2.
  • R 9 is CH 2 CH 2 OH.
  • R 20 is C(O)0(C 1 to C 6 alkyl), C(O)OH, (C 1 to C 6 alkyl)C(O)OH, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O)NHCH 2 CH 2 OH, or (C 1 to C 6
  • R 1 is NR 4 R 5 , C 1 to C 6 alkoxy, phenyl optionally substituted with C(O)0(C 1 to C 6 alkyl), C(O)OH, 0(C 1 to C 3 perfluoroalkyl), C 1 to C 6 alkoxy, halogen, CH 2 - heterocyclyl, or CH 2 CN, 5-8 membered cycloalkyl, heteroaryl, or 3-10 membered monocyclic or bicyclic heterocyclyl optionally substituted with (C 1 to C 6 alkyl)C(O)OH, Ci to C 6 alkyl, CN, C(O)OH, or (C 1 to C 6 alkyl)CN.
  • Hydrogen atoms on the same carbon atom of the cycloalkyl or heterocyclyl are optionally replaced with an optionally substituted 3-6 membered cycloalkyl or heterocyclyl to form a spirocycloalkyl or spiroheterocyclyl.
  • hydrogen atoms on the same atom of the cycloalkyl or heterocyclyl are optionally replaced with O to form an oxo substituent.
  • R 2 is phenyl or pyrazole, wherein
  • R 2 is optionally substituted with one R 19 .
  • R 19 is NR 4 R 5 , (C 1 to C 6 alkyl)NR 4 R 5 , C 1 to C 6 alkyl, C(O)NR 4 R 5 , C 3 to C 8 cycloalkyl substituted with one or more R 21 , or heterocyclyl substituted with one or more R 21 .
  • R 21 is (C 1 to C 6 alkyl)CN, (C 1 to C 6 alkyl)C(O)OH, (Ci to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O) HCH 2 CH 2 OH, or (C 1 to C 6
  • R 4 and R 5 are (a) independently selected from among H, Q to
  • R 4 and R 5 may also be (b) joined to form a 5-8 membered heterocyclyl optionally further containing one or two O, S(O)n, or NR 9 .
  • R 9 is H, OH, C 1 to C 6 hydroxyalkyl (C 1 to C 6 alkyl)C(O)OH, C(O)(C 1 to C 6 alkyl)NH 2 , C(O)(C 1 to C 6 alkyl)OH, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6 alkyl)C(O)NHCH 2 CH 2 OH, (C 1 to C 6 alkyl)C(O)N(CH 2 CH 2 OH) 2 , or C 1 to C 6 alkyl and n is 0 to 2.
  • R 9 is CH 2 CH 2 OH.
  • R 20 is C(O)0(C 1 to C 6 alkyl), C(O)OH, (C 1 to C 6 alkyl)C(O)OH, (C 1 to C 6 alkyl)C(O)NH 2 , (C 1 to C 6
  • the disclosure includes each possible combination of chiral centers within a compound, as well as all possible enantiomeric and diastereomeric mixtures thereof. All chiral, diastereomeric, and racemic forms of a structure are intended, unless the specific stereochemistry or isomeric form is specifically indicated. It is well known in the art how to prepare optically active forms, such as by resolution of racemic forms or by synthesis from optically active starting materials.
  • arylalkyloxycabonyl refers to the group (C6-C14 aryl)-(C 1 -C 6 alkyl)-O-C(O)-.
  • Terms not defined herein have the meaning commonly attributed to them by those skilled in the art.
  • Alkyl refers to a hydrocarbon chain that may be a straight chain or branched chain, containing the indicated number of carbon atoms, for example, a C1-Q2 alkyl group may have from 1 to 12 (inclusive) carbon atoms in it.
  • Examples of C 1 -C 6 alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert- butyl, isopentyl, neopentyl, and isohexyl.
  • C 1 -C 8 alkyl groups include, but are not limited to, methyl, propyl, pentyl, hexyl, heptyl, 3-methylhex-l-yl, 2,3-dimethylpent-2-yl, 3- ethylpent-l-yl, octyl, 2-methylhept-2-yl, 2,3-dimethylhex-l-yl, and 2,3,3-trimethylpent-l-yl.
  • An alkyl group can be unsubstituted or substituted with one or more of halogen, NH 2 ,
  • alkyl NH, (alkyl)(alkyl)N-, -N(alkyl)C(O)(alkyl), -NHC(O)(alkyl), -NHC(O)H, -C(O)NH 2 , - C(O)NH(alkyl), -C(O)N(alkyl)(alkyl), CN, OH, alkoxy, alkyl, C(O)OH, -C(O)0(alkyl), - C(O)(alkyl), aryl, heteroaryl, heterocyclyl, cycloalkyl, haloalkyl, aminoalkyl-, -OC(O)(alkyl), carboxyamidoalkyl-, and N0 2 .
  • Alkoxy refers to the group R-O- where R is an alkyl group, as defined above.
  • Exemplary C 1 -C 6 alkoxy groups include but are not limited to methoxy, ethoxy, n-propoxy, 1-propoxy, n-butoxy and t-butoxy.
  • An alkoxy group can be unsubstituted or substituted with one or more of halogen, OH, alkoxy, NH 2 , (alkyl)amino-, di(alkyl)amino-, (alkyl)C(O)N(Ci- C 3 alkyl)-, (alkyl)carboxyamido-, HC(O)NH-, H 2 NC(O)-, (alkyl)NHC(O)-, di(alkyl)NC(O)-, CN, C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)-, aryl, heteroaryl, cycloalkyl, haloalkyl, amino(C 1 -C 6 alkyl)-, (alkyl
  • Aryl refers to an aromatic 6 to 14 membered hydrocarbon group.
  • Ce- Ci4 aryl group include, but are not limited to, phenyl, a-naphthyl, ⁇ -naphthyl, biphenyl, anthryl, tetrahydronaphthyl, fluorenyl, indanyl, biphenylenyl, and acenanaphthyl.
  • Examples of a C6-Cioaryl group include, but are not limited to, phenyl, a-naphthyl, ⁇ -naphthyl, biphenyl, and tetrahydronaphthyl.
  • An aryl group can be unsubstituted or substituted with one or more of alkyl, halogen, haloalkyl, alkoxy, haloalkoxy, OH, hydroxyalkyl, -O- (hydroxyalkyl), -O-(alkyl)-C(O)OH, -(alkyl)-(alkoxy)-halogen, NH 2 , aminoalkyl-, dialkylamino-, C(O)OH, -C(O)0-(alkyl), -OC(O)(alkyl), -O-(alkyl)-N(alkyl)(alkyl) ,N- alkylamido-, -C(O)NH 2 , (alkyl)amido-, N0 2 , (aryl)alkyl, alkoxy, aryloxy, heteroaryloxy, (aryl)amino, (alkoxy)carbonyl-, (alkyl)amido-, (
  • bicycle or "bicyclic” as used herein refers to a molecule that features two fused rings, which rings are a cycloalkyl, heterocyclyl, or heteroaryl.
  • the rings are fused across a bond between two atoms.
  • the bicyclic moiety formed therefrom shares a bond between the rings.
  • the bicyclic moiety is formed by the fusion of two rings across a sequence of atoms of the rings to form a bridgehead.
  • a "bridge” is an unbranched chain of one or more atoms connecting two bridgeheads in a polycyclic compound.
  • the bicyclic molecule is a "spiro" or “spirocyclic” moiety.
  • the spirocyclic group is a carbocyclic or heterocyclic ring which bound through a single carbon atom of the spirocyclic moiety to a single carbon atom of a carbocyclic or heterocyclic moiety.
  • the spirocyclic group is a cycloalkyl and is bound to another cycloalkyl.
  • the spirocyclic group is a cycloalkyl and is bound to a heterocyclyl.
  • the spirocyclic group is a heterocyclyl and is bound to another heterocyclyl. In still another embodiment, the spirocyclic group is a heterocyclyl and is bound to a cycloalkyl.
  • (Aryl)alkyl refers to an alkyl group, as defined above, wherein one or more of the alkyl group's hydrogen atoms has been replaced with an aryl group as defined above.
  • (C6-C14 aryl)alkyl- moieties include benzyl, benzhydryl, 1-phenylethyl, 2-phenylethyl, 3- phenylpropyl, 2-phenylpropyl, 1-naphthylmethyl, 2-naphthylmethyl and the like.
  • An (aryl)alkyl group can be unsubstituted or substituted with one or more of of halogen, CN, NH 2 , OH, (alkyl)amino-, di(alkyl)amino-, (alkyl)C(O)N(alkyl)-, (alkyl)carboxyamido-, HC(O)NH-, H 2 NC(O)-, (alkyl)NHC(O)-, di(alkyl)NC(O)-, CN, OH, alkoxy, alkyl, C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)-, aryl, heteroaryl, cycloalkyl, haloalkyl, amino(alkyl)-, (alkyl)carboxyl-, carboxyamidoalkyl-, or N0 2 .
  • (Alkoxy)carbonyl- refers to the group alkyl-O-C(O)-.
  • Exemplary (C 1 -C 6 alkoxy)carbonyl- groups include but are not limited to methoxy, ethoxy, n-propoxy, 1- propoxy, n-butoxy and t-butoxy.
  • An (alkoxy)carbonyl group can be unsubstituted or substituted with one or more of halogen, OH, NH 2 , (alkyl)amino-, di(alkyl)amino-,
  • alkyl)C(O)N(alkyl)- (alkyl)carboxyamido-, HC(O)NH-, H 2 NC(O)-, (alkyl)NHC(O)-, di(alkyl)NC(O)-, CN, alkoxy, C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)-, aryl, heteroaryl, cycloalkyl, haloalkyl, amino(alkyl)-, (alkyl)carboxyl-, carboxyamidoalkyl-, orN0 2 .
  • (Alkyl)amido- refers to a -C(O)NH- group in which the nitrogen atom of said group is attached to a C 1 -C 6 alkyl group, as defined above.
  • Representative examples of a (C 1 -C 6 alkyl)amido- group include, but are not limited to, -C(O)NHCH 3 , -C(O)NHCH 2 CH 3 , -C(O)NHCH 2 CH 2 CH 3 , -C(O)NHCH 2 CH 2 CH 2 CH 3 , -C(O)NHCH 2 CH 2 CH 2 CH 2 CH 3 , -C(O)NHCH(CH 3 ) 2 , -C(O)NHCH 2 CH(CH 3 ) 2 , -C(O)NHCH(CH 3 )CH 2 CH 3 , -C(O)NH- C(CH 3 ) 3 and -C(O)NHCH 2 C(CH 3 ) 3 .
  • (Alkyl)amino- refers to an -NH group, the nitrogen atom of said group being attached to a alkyl group, as defined above.
  • Representative examples of an (C 1 -C 6 alkyl)amino- group include, but are not limited to CH 3 NH-, CH 3 CH 2 NH-, CH 3 CH 2 CH 2 NH-, CH 3 CH 2 CH 2 CH 2 NH-, (CH 3 ) 2 CHNH-, (CH 3 ) 2 CHCH 2 NH-, CH 3 CH 2 CH(CH 3 )NH- and (CH 3 ) 3 CNH-.
  • An (alkyl)amino group can be unsubstituted or substituted on the alkyl moiety with one or more of halogen, NH 2 , (alkyl)amino-, di(alkyl)amino-, (alkyl)C(O)N(alkyl)-, (alkyl)carboxyamido-, HC(O)NH-, H 2 NC(O)-, (alkyl)NHC(O)-, di(alkyl)NC(O)-, CN, OH, alkoxy, alkyl, C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)-, aryl, heteroaryl, cycloalkyl, haloalkyl, amino(alkyl)-, (alkyl)carboxyl-, carboxyamidoalkyl-, or N0 2 .
  • halogen NH 2 , (alkyl)amino-, di(al
  • Aminoalkyl- refers to an alkyl group, as defined above, wherein one or more of the alkyl group's hydrogen atoms has been replaced with -NH 2 ; one or both H of the NH 2 may be replaced by a substituent.
  • Alkylcarboxyl- refers to an alkyl group, defined above that is attached to the parent structure through the oxygen atom of a carboxyl (C(O)-O-) functionality. Examples of (C 1 -C 6 alkyl)carboxyl- include acetoxy, propionoxy, propylcarboxyl, and isopentylcarboxyl.
  • (Alkyl)carboxyamido- refers to a -NHC(O)- group in which the carbonyl carbon atom of said group is attached to a C 1 -C 6 alkyl group, as defined above.
  • Representative examples of a (C 1 -C 6 alkyl)carboxyamido- group include, but are not limited to,
  • (Aryl)amino refers to a radical of formula (aryl)-NH-, wherein aryl is as defined above.
  • (Aryl)oxy refers to the group Ar-O- where Ar is an aryl group, as defined above.
  • Cycloalkyl refers to a non-aromatic, saturated, partially saturated, monocyclic, bicyclic or polycyclic hydrocarbon 3 to 12 membered ring system.
  • Representative examples of a C 3 -Ci 2 cycloalkyl include, but are not limited to, cyclopropyl, cyclopentyl, cycloheptyl, cyclooctyl, decahydronaphthalen-l-yl, octahydro-lH-inden-2-yl, decahydro-lH- benzo[7]annulen-2-yl, and dodecahydros-indacen-4-yl.
  • C 3 -Cio cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, decahydronaphthalen-l-yl, and octahydro-lH-inden-2-yl.
  • C 3 -C 8 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and octahydropentalen-2-yl.
  • a cycloalkyl can be unsubstituted or substituted with one or more of halogen, NH 2 , (alkyl)NH, (alkyl)(alkyl)N-, -N(alkyl)C(O)(alkyl), -NHC(O)(alkyl), -NHC(O)H, -C(O)NH 2 ,
  • Halo or "halogen” refers to -F, -CI, -Br and -I.
  • C 1 -C 6 haloalkyl refers to a C 1 -C 6 alkyl group, as defined above, wherein one or more of the C 1 -C 6 alkyl group's hydrogen atoms has been replaced with F, CI, Br, or I. Each substitution can be independently selected from F, CI, Br, or I.
  • an C 1 -C 6 haloalkyl- group include, but are not limited to, -CH 2 F, -CC1 3 , -CF 3 , CH 2 CF 3 , -CH 2 C1, -CH 2 CH 2 Br, -CH 2 CH 2 I, -CH 2 CH 2 CH 2 F, -CH 2 CH 2 CH 2 C1, -CH 2 CH 2 CH 2 CH 2 Br, -CH2CH2CH2I, -CH 2 CH2CH2CH 2 CH2Br, -CH2CH2CH2CH2I, -CH 2 CH(Br)CH 3 , -CH 2 CH(C1)CH 2 CH3, -CH(F)CH 2 CH 3 and -C(CH 3 )2(CH 2 C1).
  • Heteroaryl refers to a monocyclic, bicyclic, or polycyclic aromatic ring system containing at least one ring atom selected from the heteroatoms oxygen, sulfur and nitrogen.
  • C1-C9 heteroaryl groups include furan, thiophene, indole, azaindole, oxazole, thiazole, isoxazole, isothiazole, imidazole, N-methylimidazole, pyridine, pyrimidine, pyrazine, pyrrole, N-methylpyrrole, pyrazole, N-methylpyrazole, 1,3,4-oxadiazole, 1,2,4- triazole, l-methyl-l,2,4-triazole, lH-tetrazole, 1 -methyltetrazole, benzoxazole,
  • Bicyclic C1-C9 hetroaryl groups include those where a phenyl, pyridine, pyrimidine or pyridazine ring is fused to a 5 or 6-membered monocyclic heteroaryl ring having one or two nitrogen atoms in the ring, one nitrogen atom together with either one oxygen or one sulfur atom in the ring, or one O or S ring atom.
  • Examples of monocyclic C1-C4 heteroaryl groups include 2H-tetrazole, 3H-l,2,4-triazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, imidazole, and pyrrole.
  • a heteroaryl group can be unsubstituted or substituted with one or more of C 1 -C 6 alkyl, halogen, haloalkyl, OH, CN, hydroxyalkyl, N3 ⁇ 4, aminoalkyl-, dialkylamino-, C(O)OH, -C(O)0-(alkyl), -OC(O)(alkyl), N-alkylamido-, -C(O)NH 2 , (alkyl)amido-, -N0 2 , (aryl)alkyl, alkoxy, aryloxy, heteroaryloxy, (aryl)amino, (alkoxy)carbonyl-, (alkyl)amido-, (alkyl)amino, aminoalkyl-, alkylcarboxyl-, (alkyl)carboxyamido-, (aryl)alkyl-, (aryl)amino-, cycloalkenyl, di(alkyl
  • Heterocycle refers to monocyclic, bicyclic, polycyclic, or bridged head molecules in which at least one ring atom is a heteroatom.
  • a heterocycle maybe saturated or partially saturated.
  • Exemplary C1-C9 heterocyclyl groups include but are not limited to aziridine, oxirane, oxirene, thiirane, pyrroline, pyrrolidine, dihydrofuran, tetrahydro furan, dihydrothiophene, tetrahydrothiophene, dithiolane, piperidine, 1,2,3,6- tetrahydropyridine-l-yl, tetrahydropyran, pyran, thiane, thiine, piperazine, azepane, diazepane, oxazine, 5,6-dihydro-4H-l,3-oxazin-2-yl, 2,5-diazabicyclo[2.2.1]heptane,
  • Ci heterocyclyl radicals would include but are not limited to oxaziranyl, diaziridinyl, and diazirinyl
  • C 2 heterocyclyl radicals include but are not limited to aziridinyl, oxiranyl, and diazetidinyl
  • C heterocyclyl radicals include but are not limited to azecanyl, tetrahydroquinolinyl, and perhydroisoquinolinyl.
  • a heterocyclyl group can be unsubstituted or substituted with one or more of alkyl, halogen, alkoxy, haloalkyl, OH, hydroxyalkyl, -C(O)-(hydroxyalkyl), NH 2 , aminoalkyl-, dialkylamino-, C(O)OH, -C(O)0- (alkyl), -OC(O)(alkyl), N-alkylamido-, -C(O)NH 2 , (alkyl)amido-, -C(O)-(alkyl)-CN, (alkyl)- CN, or N0 2 .
  • Heterocyclyl(alkyl)- refers to an alkyl group, as defined above, wherein one or more of the alkyl group's hydrogen atoms has been replaced with a heterocycle group as defined above.
  • Heterocyclyl( C 1 -C 6 alkyl)- moieties include 1 -piperazinylethyl, 4-morpholinylpropyl, 6-piperazinylhexyl, and the like.
  • a heterocyclyl(alkyl) group can be unsubstituted or substituted with one or more of halogen, NH 2 , (alkyl)amino-, di(alkyl)amino-,
  • alkyl)C(O)N(alkyl)- (alkyl)carboxyamido-, HC(O)NH-, H 2 NC(O)-, (alkyl)NHC(O)-, di(alkyl)NC(O)-, CN, OH, alkoxy, alkyl, C(O)OH, (alkoxy)carbonyl-, (alkyl)C(O)-, 4- to 7- membered monocyclic heterocycle, aryl, heteroaryl, or cycloalkyl.
  • Heteroaryl(alkyl) refers to a heteroaryl which is attached to an alkyl group and the heteroaryl is defined above.
  • Hydroalkyl refers to a alkyl group, as defined above, wherein one or more of the alkyl group's hydrogen atoms has been replaced with OH groups.
  • C 1 -C 6 hydroxyalkyl moieties include, for example, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, -CH 2 CH(OH)CH 2 OH, -CH 2 CH(OH)CH 3 , -CH(CH 3 )CH 2 OH and higher homologs.
  • Perfluoroalkyl- refers to alkyl group, defined above, having two or more fluorine atoms. Examples of a C 1 -C 6 perfluoroalkyl- group include CF3, CH 2 CF3, CF 2 CF3 and CH(CF 3 ) 2 .
  • a "subject” is a mammal, e.g. , a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or non-human primate, such as a monkey, chimpanzee, baboon or gorilla.
  • salts include but are not limited to, e.g., water-soluble and water-insoluble salts, such as the acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, bromide, butyrate, calcium, chloride, choline, citrate, edisylate (camphorsulfonate), fumarate, gluconate, glucuronate, glutamate, hydrobromide, hydrochloride, lauryl sulfate, malate, maleate, mandelate, mesylate, palmitate, pantothenate, phosphate, potassium, propionate, p-toluenesulfonate, salicylate, sodium, stearate, succinate, and sulfate salts.
  • water-soluble and water-insoluble salts such as the acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, bromide,
  • ACN is acetonitrile
  • DMSO dimethylsulfoxide
  • DMF is N,N-dimethylformamide
  • DMF.DMA dimethylformamide dimethylacetal
  • TFA trifluroroacetic acid
  • mCPBA meta- chloroperbenzoic acid
  • RT room temperature
  • THF is tetrahydrofuran
  • NMP N- methyl pyrrolidinone
  • Scheme 1 provides the synthesis of compounds of Formula (I).
  • Ethyl acetoacetate 1 is converted to the corresponding bis(methylthio)methylene derivative 2 using carbon disulfide, an organic or inorganic base such as K2CO3 and a alkylating agent.
  • the alkylating agent is an alkyl iodide, alkyl triflate, or alkyl sulfonate.
  • the alkylating agent is a methylating agent.
  • the alkylating agent is methyl iodide. Reaction of 2 with an R 1 -optionally substituted amidine hydrochloride in the presence of a base results in pyrimidine 3.
  • the base utilized to form pyrimidine 3 is EtsN or Hiinig's base.
  • the alkyl group on the pyrimidine group of compound 3 is then oxidized using an oxidizing agent. In one embodiment, the oxidation is performed using Se0 2 .
  • the resulting pyrimidine aldehyde 4 is converted to pyrimido-pyridazinone 5 using hydrazine hydrate or hydrazine hydrochloride.
  • the methyl thio group in compound 5 is oxidized to a methane sulfonyl using meta-chloroperoxybenzoic acid (mCPBA) or hydrogen peroxide/acetic acid.
  • mCPBA meta-chloroperoxybenzoic acid
  • the methane sulfonyl group of compound 6 is replaced with an R 2 -substituted aniline to provide compound (I).
  • the R 2 -substituted aniline is an aryl or heteroaryl substituted aniline.
  • Scheme 2 provides the synthesis of compound IB which are encompassed by the structure of Formula (I).
  • ethyl acetoacetate 1 is converted to the corresponding bis(methylthio)methylene derivative 2 using carbon disulfide, K2CO3 and methyl iodide.
  • Reaction of 2 with an R-substituted benzamidine hydrochloride in the presence of Et3N results inpyrimidine 3a.
  • the methyl group bound to the C-atom of pyrimidine 3a is then oxidized using Se0 2 .
  • the resulting pyrimidine aldehyde 4a is converted to pyrimido-pyridazinone 5a using hydrazine hydrate or hydrazine hydrochloride.
  • Scheme 3 provides the synthesis of compounds IC which are encompassed by the structure of Formula (I).
  • Treatment of ethyl chloroformate 7 with ammonium thiocyanate results in the production of ethyl thiocyanato formate 8 which upon treatment with ethyl 3- amino crotanoate results in compound 9.
  • Compound 9 is cyclized to compound 10 by treatment with an organic or inorganic base.
  • the organic or inorganic base is a strong base.
  • the strong base is a tertiary organic base.
  • the strong base is aqueous Et3N.
  • the dichloro pyrimidine 11 is obtained by treating compound 10 with a chlorinating agent.
  • the chlorinating agent is POCI3.
  • this transformation can also be carried out by using other chlorinating agents such as PCI5, SOCl 2 in the presence of an organic base such as TEA, tributyl amine, and ⁇ , ⁇ -dimethylaniline.
  • the 4-position of dichloropyrimidine 11 is then substituted by reaction with an optionally substituted (R 2 ) aniline to afford compound 12.
  • the 2-position of pyrimidine 12 is then R ⁇ -substituted using coupling agents such as boronic acids or boronic ester reagents to provide compound 13.
  • the methyl group at position 4 of pyrimidine 13 is then oxidized using an oxidizing agent such as Se0 2 to provide compound 14.
  • the resulting pyrimidine aldehyde 14 is converted to pyrimido-pyridazinone IC using hydrazine hydrate.
  • Scheme 4 provides the synthesis of compounds ID which are encompassed by the structure of Formula (I).
  • Treatment of ethyl chloroformate 7 with ammonium thiocyanate results in ethyl thiocyanato formate 8 which upon treatment with ethyl 3-amino crotanoate results in compound 9.
  • Compound 9 is cyclized to compound 10 as described in Scheme 3, i.e., by treatment with aqueous Et3N.
  • the dichloro pyrimidine compound 11 is obtained by treating compound 10 with POCI3.
  • the 4-position of dichloropyrimidine 11 is then substituted by reaction with an optionally R-substituted aniline to afford compound 12a.
  • the 2-position of pyrimidine compound 12a is then substituted with an R'-substituted aryl or heteroaryl group using a boronic acid or an boronic ester reagent to provide compound 13a.
  • the boronic acid is (R'-aryl)-B(OH)2 or (R'-heteroaryl)-B(OH)2.
  • the methyl at position-4 on pyrimidine 13a is then oxidized using Se0 2 .
  • the resulting pyrimidine aldehyde 14a is converted to pyrimido-pyridazinone ID using hydrazine hydrate.
  • Scheme 5 provides the synthesis of compound IE which are encompassed by the structure of Formula (I).
  • Compound 12 is reacted with an optionally substituted amine (NHR 4 R 5 ) to provide compound 15.
  • the methyl group at the 4-position of compound 15 is reacted with DMF.DMA to provide compound 16.
  • Compound 17 is obtained by oxidative cleavage of the olefin of compound 16. In one embodiment, oxidative cleavage is performed using NaI0 4 .
  • pyrimido-pyridazinone IE is obtained by cyclizing the aldehyde 17. In one embodiment, the cyclizaton is performed using hydrazine, hydrazine hydrate or hydrazine hydrochloride, as described previously.
  • Scheme 6 provides the synthesis of compounds IF which are encompassed by the structure of Formula (I).
  • Compound 12a is reacted with optionally substituted amines to result in 15a.
  • the methyl group of 15a is reacted with DMF.DMA to give compound 16a.
  • the aldehyde 17a is obtained by the oxidative cleavage of the olefin in 16a. In one embodiment, the oxidative cleavage is performed with NaI0 4 . Finally, the pyrimido- pyridazinone IF is obtained by cyclizing aldehyde 17a. In one embodiment, cyclization is performed using with hydrazine.
  • Scheme 7 provides the synthesis of compound 1G which are encompassed by the structure of Formula (I).
  • Compound 10 is reacted with an alkylating agent to the S-methyl compound 18.
  • the reacted is performed under basic conditions.
  • the alkylating agent is methyl iodide, ethyl iodide, propyl iodide, dimethylsulfate, among others.
  • Compound 19 is obtained by chlorinating compound 18.
  • compound 18 is chlorinated using POCI3.
  • Compound 19 is then NR 4 R 5 substituted with an optionally substituted amine to provide compound 20 [using NHR 4 R 5 ?].
  • the methyl group of compound 20 is reacted with DMF.DMA to give compound 21.
  • the aldehyde 22 is obtained by the oxidative cleavage of the olefin group in compound 21.
  • the oxidative cleavage is performed with NaI0 4 .
  • compound 20 may directly converted to the pyrimidine aldehyde 22 by oxidizing the methyl group using Se0 2 or a combination of C0 2 , t-butyl hydroperoxide, and an alcohol such as Ci to Ce alkyl)H 2 OH.
  • the resulting pyrimidine aldehyde 22 is converted to pyrimido-pyridazinone 23 using hydrazine hydrate or hydrazine hydrochloride.
  • the methyl thio group in compound 23 is oxidized to a methane sulfonyl group.
  • compound 23 is reacted with meta-chloroperoxybenzoic acid (mCPBA) or hydrogen peroxide/acetic acid.
  • mCPBA meta-chloroperoxybenzoic acid
  • the methane sulfonyl group of compound 24 is replaced with suitably substituted aniline to provide compound 1G.
  • Scheme 8 provides the synthesis of compound 1H which is encompassed by the structure of Formula (I).
  • compound 19 is coupled with an optionally substituted boronic acid or boronic ester to give compound 25.
  • the coupling is performed in the presence of a coupling agent such as Pd(PPh3) 4 or PdCl 2 (PPh3) 2 .
  • the methyl group in compound 25 is then oxidized to the corresponding aldehyde.
  • the oxidation is performed using Se0 2 or a combination of C0 2 , t-butyl hydroperoxide, and an alcohol such as C 1 to C 6 alkyl)H 2 OH to give compound 26.
  • Compound 26 is converted to pyrimido-pyridazinone 27 using hydrazine hydrate or hydrazine hydrochloride.
  • the methyl thio group in compound 27 is then oxidized to a methane sulfonyl group.
  • the oxidation is performed using mCPBA or hydrogen peroxide/acetic acid.
  • the methane sulfonyl group of compound 28 is replaced with suitably substituted aniline to provide compound 1H.
  • Pharmaceutical combinations of the disclosure comprise a compound of Formula (I) optionally with other pharmaceutically inert or inactive ingredients.
  • the pharmaceutically inert or inactive ingredient is one or more pharmaceutically acceptable carrier or excipient.
  • the present disclosure also contemplates combining the compound of Formula (I) with one or more therapeutic agents, i.e., active ingredients, as described below.
  • a compound of Formula (I) is combined with one or more inert/inactive ingredients and one or more therapeutic agents.
  • the pharmaceutical combinations of the disclosure contain an amount of a compound of Formula (I) that is effective for treating inflammation in a subject.
  • the dosage of the compound of Formula (I) to achieve a therapeutic effect will depend on factors such as the formulation, pharmacological potency of the drug, age, weight and sex of the patient, condition being treated, severity of the patient's symptoms, specific compound of Formula
  • the treatment and dosage of the compound of Formula (I) maybe administered in unit dosage form and that one skilled in the art would adjust the unit dosage form accordingly to reflect the relative level of activity.
  • the decision as to the particular dosage to be employed (and the number of times to be administered per day) is within the discretion of the ordinarily-skilled physician, and maybe varied by titration of the dosage to the particular circumstances to produce the desired therapeutic effect.
  • the therapeutically effective amount is about 0.0001% to about 25% w/w. In another embodiment, the therapeutically effective amount is less than about 20% w/w, about 15% w/w, about 10% w/w, about 5% w/w, or about 1% w/w. In another embodiment, the therapeutically effective amount is about 0.0001% to about 10% w/w. In a further embodiment, the therapeutically effective amount is about 0.005 to about 5% w/w. In yet another embodiment, the therapeutically effective amount is about 0.01 to about 5% w/w.
  • the therapeutically effective amount is about 0.01% w/w, about 0.05% w/w, about 0.1 % w/w, about 0.2 % w/w, about 0.3% w/w, about 0.4% w/w, about
  • the therapeutically effective amounts maybe provided on regular schedule, i.e., on a less than daily, weekly, monthly, or yearly basis or on an irregular schedule with varying administration days, weeks, months, etc.
  • the therapeutically effective amount to be administered may vary.
  • the therapeutically effective amount for the first dose is higher than the therapeutically effective amount for one or more of the subsequent doses.
  • the therapeutically effective amount for the first dose is lower than the therapeutically effective amount for one or more of the subsequent doses.
  • Equivalent dosages may be administered over various time periods including, but not limited to, about every 2 hours, about every 6 hours, about every 8 hours, about every 12 hours, about every 24 hours, about every 36 hours, about every 48 hours, about every 72 hours, about every week, about every 2 weeks, about every 3 weeks, about every month, about every 2 months, about every 3 months and about every 6 months.
  • the number and frequency of dosages corresponding to a completed course of therapy will be determined according to the judgment of a health-care practitioner.
  • the therapeutically effective amounts described herein refer to total amounts administered for a given time period; that is, if more than one compound of Formula (I) is administered, the therapeutically effective amounts correspond to the total amount administered.
  • the compound of Formula (I) maybe administered by any route, taking into consideration the specific condition for which it has been selected.
  • the compounds of Formula (I) may be delivered orally, by injection (including intravascularly, e.g ,
  • the compound of Formula (I) may be administered by injection, transdermally or topically.
  • the compound of Formula (I) maybe administered topically to the eye, e.g., as solutions, suspensions or ointments.
  • ophthalmically compatible carriers which may be used include, without limitation, an aqueous solution, such as saline solution, oil solution or ointments containing ophthalmically compatible preservatives, surfactants, buffers, and viscosity regulators. These compositions may also contain stabilizing agents, antibacterial agents, and maybe manufactured in different dosage units, suitable for ocular administration. Drug inserts, either soluble or insoluble, may also be used.
  • the compound of Formula (I) maybe administered by injection.
  • Solutions for injection or infusion may be prepared as aqueous solutions.
  • the compound of Formula (I) is present in a concentration of about 0.001 ⁇ g mL to 1 mg/mL, or this amount maybe adjusted higher or lower as needed.
  • These solutions may also contain stabilizing agents, antibacterial agents, buffers and maybe manufactured in different dosage unit ampoules or bottles.
  • the compound of Formula (I) maybe administered rectally.
  • Dosage units for rectal administration may be prepared in the form of ointments or suppositories, which contain the compound of Formula (I) in a mixture with a neutral fat base, or they may be prepared in the form of gelatin-rectal capsules that contain the compound of Formula (I) in a mixture with, e.g., a vegetable oil or paraffin oil.
  • Ointments, suppositories or creams containing at least one compound of Formula (I) are useful for the treatment of hemorrhoids.
  • the compound of Formula (I) maybe administered orally.
  • Dosage units for oral administration include, without limitation, tablets, caplets, capsules, powders, suspensions, microcapsules, dispersible powder, granules, suspensions, syrups,. elixirs, and aerosols, which contain the compound of Formula (I) optionally with one or more excipient.
  • the compositions are compressed into a tablet or caplet.
  • the tablet or caplet maybe administered to the subject.
  • the tablet or caplet may be added to a capsule.
  • the composition containing the compound of Formula (I) is added directly to a capsule.
  • the capsule includes hydroxypropyl methylcellulose, hypromellose capsule, or a hard shell gelatin capsule.
  • the tablets or caplets are optionally film-coated using film-coatings known to those of skill in the art.
  • the film-coating is selected from among polymers such as, without limitation,
  • hydroxypropylmethylcellulose ethyl cellulose
  • polyvinyl alcohols and combinations thereof.
  • the compound of Formula (I) may also be administered in the presence of one or more pharmaceutical carriers that are physiologically compatible.
  • the amount of the pharmaceutical carrier(s) is determined by the solubility and chemical nature of the compound of Formula (I), chosen route of administration, and standard pharmacological practice.
  • the carriers may be in dry (solid) or liquid form and must be pharmaceutically acceptable.
  • Liquid pharmaceutical compositions are typically sterile solutions or suspensions. When liquid carriers are utilized for parenteral administration, they are desirably sterile liquids. Liquid carriers are typically utilized in preparing solutions, suspensions, emulsions, syrups and elixirs. A variety of suitable liquid carriers is known and may be readily selected by one of skill in the art.
  • Such carriers may include, .e.g., dimethylsulfoxide (DMSO), saline, buffered saline, cyclodextrin,
  • the compound of Formula (I) is dissolved a liquid carrier.
  • the compound of Formula (I) is suspended in a liquid carrier.
  • a suitable liquid carrier depending on the route of administration.
  • the compound of Formula (I) may alternatively be formulated in a solid carrier of which a variety of solid carriers and excipients are known to those of skill in the art.
  • the composition maybe compacted into a unit dose form, i.e., tablet or cap let.
  • the composition maybe added to unit dose form, i.e., a capsule.
  • the composition maybe formulated for administration as a powder.
  • the solid carrier may perform a variety of functions, i.e., may perform the functions of two or more of the excipients described below.
  • a solid carrier may also act as a flavoring agent, lubricant, solubilizer, suspending agent, filler, glidant, compression aid, binder, disintegrant, or encapsulating material.
  • compositions may also be sub-divided to contain appropriate quantities of the compound of Formula (I).
  • the unit dosage can be packaged compositions, e.g., packeted powders, vials, ampoules, prefilled syringes or sachets containing liquids.
  • excipients which may be combined with one or more compound of
  • Formula (I) include, without limitation, adjuvants, antioxidants, binders, buffers, coatings, coloring agents, compression aids, diluents, disintegrants, emulsifiers (e g. , polyoxyethylene fatty acid esters), emollients, encapsulating materials, fillers, flavoring agents, glidants, granulating agents, lubricants, metal chelators, osmo-regulators, pH adjustors (e.g., sodium hydroxide), preservatives, solubilizers, sorbents, stabilizing agents, sweeteners (such as saccharin), surfactants, suspending agents, syrups, thickening agents (e.g.,
  • penetration enhancers e.g., hydroxypolyethoxydodecane, DMSO, DMAC, DDM, etc
  • viscosity regulators such as polymers to increase viscosity
  • compositions maybe utilized as inhalants.
  • compositions maybe prepared as fluid unit doses using a compound of Formula (I) and a vehicle for delivery by an atomizing spray pump or by dry powder for insufflation.
  • compositions maybe utilized as aerosols, i.e., oral or intranasal.
  • aerosols i.e., oral or intranasal.
  • the compositions are formulated for use in a pressurized aerosol container together with a gaseous or liquefied propellant, e.g. ,
  • dichlorodifluoromethane carbon dioxide, nitrogen, propane, and the like. Also provided is the delivery of a metered dose in one or more actuations.
  • compositions may be administered by a modified-release delivery device.
  • Modified-release refers to delivery of a compound of Formula (I) which is controlled, for example over a period of at least about 8 hours (e.g., extended delivery) to at least about 12 hours (e.g., sustained delivery). Such devices may also permit immediate release (e.g., therapeutic levels achieved in under about 1 hour, or in less than about 2 hours).
  • suitable modified-release delivery devices For use in such modified-release delivery devices, the compound of Formula (I) is formulated as described herein.
  • the compounds of Formula (I) are combined with other medications or therapeutic agents in a single composition.
  • the present disclosure is not so limited.
  • the compounds of Formula (I) may be administered in one or more separate formulations from other compounds of Formula (I), or other medications or therapeutic agents as described below.
  • agents typically used to treat inflammation may be used in conjunction with a combination of the disclosure in the methods, compositions, and kits described herein.
  • agents include, but are not limited to, non-steroidal anti-inflammatory drugs.
  • the compound of Formula (I) maybe combined with glucose or dextrose when utilized for infusion or as a regional analgesic or anti-pruritic.
  • the compound of Formula (I) maybe combined with thickening agents to form a jelly, or may also contain penetration enhancers, for use in topical or dermal applications such as for urogenital topical procedures.
  • the compound of Formula (I) maybe formulated as an ointment for administration to accessible mucous membranes.
  • kits or packages of pharmaceutical formulations containing the compounds of Formula (I) or compositions described herein are also provided herein.
  • the kits maybe organized to indicate a single formulation or combination of formulations to be taken at each desired time.
  • the kit contains packaging or a container with the compound of Formula (I) formulated for the desired delivery route.
  • the kit contains instructions on dosing and an insert regarding the compound of Formula (I).
  • the kit may further contain instructions for monitoring local or circulating levels of product and materials for performing such assays including, e.g., reagents, well plates, containers, markers or labels, and the like.
  • Such kits are readily packaged in a manner suitable for treatment of a desired indication.
  • the kit may also contain instructions for use of an oral dosage form such as a pill, capsule, patch, spray pump or other delivery device.
  • suitable components to include in such kits will be readily apparent to one of skill in the art, taking into consideration the desired indication and the delivery route.
  • a package or kit can include the compound of Formula (I) in each dosage unit (e.g., solution, lotion, tablet, pill, drug-eluting patch or other unit described above or utilized in drug delivery), and optionally instructions for administering the doses less-than-daily, daily, weekly, or monthly, for a predetermined length of time or as prescribed.
  • a package or kit can include placebos during periods when the compound of Formula (I) is not delivered.
  • a package or kit may contain a sequence of dosage units which provide the desired variability.
  • the package has indicators for each period.
  • the package is a foil or blister package, labeled ampoule, vial or bottle.
  • the packaging means of a kit may itself be geared for administration, such as an inhalant, syringe, pipette, eye dropper, or other such apparatus, from which the formulation may be applied to an affected area of the body, such as the lungs, injected into a subject, or even applied to and mixed with the other components of the kit.
  • kits also may be provided in dried or lyophilized forms.
  • reagents or components are provided as a dried form, reconstitution generally is by the addition of a suitable solvent. It is envisioned that the solvent also may be provided in another package.
  • kits of the present disclosure also will typically include a means for containing the vials or other suitable packaging means in close confinement for commercial sale such as, e.g. , injection or blow-molded plastic containers into which the desired vials are retained.
  • a means for containing the vials or other suitable packaging means in close confinement for commercial sale such as, e.g. , injection or blow-molded plastic containers into which the desired vials are retained.
  • the kits also may include, or be packaged with a separate instrument for assisting with the injection/administration or placement of the composition within the body of an animal.
  • a separate instrument maybe an inhalant, syringe, pipette, forceps, measuring spoon, eye dropper or any such medically approved delivery means.
  • a kit in one embodiment, contains a compound of Formula (I).
  • the compound of Formula (I) may be in the presence or absence of one or more of the carriers or excipients described above.
  • the kit may optionally contain instructions for administering the compound of Formula (I) to a subject having inflammation.
  • a kit in a further embodiment, contains a compound of Formula (I) in a second dosage unit, and one or more of the carriers or excipients described above in a third dosage unit.
  • the kit may optionally contain instructions for administering the compound of
  • Formula (I) to a subject having inflammation.
  • inflammation as used herein includes all types of inflammation.
  • the inflammation may be acute or chronic.
  • the inflammation may be nociceptive,
  • the inflammation maybe from a migraine, gynecological condition, pre-labor or labor, stroke, surgery, neuralgia, sickle cell, interstitial cystitis, urological condition (such as urethritis), dental work/injury, or headache, among other.
  • Inflammation may also occur in patients with cancer, which may be due to multiple causes, such as nerve compression and mechanical forces resulting from tissue distension as a consequence of invasion by a tumor and tumor metastasis into bone or other tissues.
  • inflammation results from neuropathy, such as post-herpetic neuralgia.
  • the inflammation results from a surgery or procedure.
  • the inflammation results from an infection, cancer, colitis, cystitis, irritable bowel syndrome, or idiopathic neuropathy
  • Somatic inflammation includes inflammation in bone, joint, muscle, skin, or connective tissue.
  • Central inflammation includes inflammation arising as a consequence of brain trauma, stroke, or spinal cord injury.
  • Viceral inflammation includes inflammation in visceral organs, such as the respiratory or gastrointestinal tract and pancreas, the urinary tract and reproductive organs. In one embodiment, visceral inflammation results from tumor involvement of the organ capsule. In another embodiment, visceral inflammation from obstruction of hollow viscus.
  • Idiopathic inflammation refers to inflammation which has no underlying cause or refers to inflammation caused by condition which remains undiagnosed.
  • Disfunctional inflammation refers to inflammation which occurs in the absence of a noxious stimulus, tissue damage or a lesion to the nervous system.
  • dysfunctional inflammation results from rheumatologic conditions such as arthritis and fibromyalgia, tension type headache, irritable bowel disorders and erythermalgia.
  • Nociceptive inflammation includes inflammation caused by noxious stimuli that threaten to or actually injure body tissues.
  • nociceptive inflammation results from a cut, bruise, bone fracture, crush injury, burn, trauma, surgery, labor, sprain, bump, injection, dental procedure, skin biopsy, or obstruction.
  • nociceptive inflammation is located in the skin, musculoskeletal system, or internal organs.
  • Neurotoxic inflammation is inflammation due to abnormal processing of sensory input by the peripheral or central nervous system consequent on a lesion to these systems.
  • neuropathic inflammation is chronic and non-malignant.
  • neuropathic inflammation is due to trauma, surgery, herniation of an intervertebral disk, spinal cord injury, diabetes, infection with herpes zoster (shingles), HIV/AIDS, late-stage cancer, amputation (such as mastectomy), carpal tunnel syndrome, chronic alcohol use, exposure to radiation, and as an unintended side-effect of neurotoxic treatment agents, such as certain anti-HIV and chemotherapeutic drugs.
  • Procedural inflammation includes refers to inflammation arising from a medical procedure.
  • the medical procedure may include any type of medical, dental or surgical procedure.
  • the procedural inflammation is postoperative.
  • the inflammation is associated with an injection, draining an abscess, surgery, dermatological, dental procedure, ophthalmic procedure, arthroscopy and use of other medical instrumentation, and/or cosmetic surgery.
  • a “migraine” is a type of headache, typically defined clinically as being caused by activation of sensory fibers innervating the meninges of the brain.
  • treat means to include therapy utilized to remedy a health problem or condition in a patient or subject.
  • the health problem or condition may be eliminated permanently or for a short period of time.
  • the health problem or condition may be prevented.
  • the severity of the health problem or condition, or of one or more symptoms characteristic of the health problem or condition may be lessened permanently, or for a short period of time.
  • the effectiveness of a treatment of inflammation can be determined using any standard inflammation index, such as those described herein, or can be determined based on the patient's subjective inflammation assessment. A patient is considered “treated” if there is a reported reduction in inflammation, or a reduced reaction to stimuli that should cause inflammation.
  • the treatment methods described herein include administering a compound of Formula (I) to a patient. Additional, optional agents, such as those described above for use in the combination, may be administered to the patient prior to, concurrently with, or subsequent to the compound of Formula (I).
  • Ethyl 2-hydroxy-4-mercapto-6-methylpyrimidine-5-carboxylate (1.4 g, 6.54 mmol) was taken up in POCI3 (13.67 mL) at 0°C, tri-n-butylamine was added and the mixture heated to 90°C for 5 hours. The mixture was cooled, poured slowly on to crushed ice and extracted with dichloromethane. The organic layer was washed with water, dried over Na 2 S0 4 and concentrated under vacuum.
  • Example 340 Inhibition of enzymatic Syk kinase activity
  • the objective of this assay was to examine by radiometric method the ability of compounds to inhibit Syk kinase enzyme.
  • Spleen tyrosine kinase is a cytosolic protein tyrosine kinase that plays a crucial role in inflammatory and allergic responses. Syk triggers IgE and IgG receptor mediated signaling in mast cells, basophils, and macrophages leading to degranulation and cytokine release. Abnormal function of Syk has also been implicated in several instances of hematopoietic malignancies.
  • Syk is capable of phosphorylating substrates such as VAV, LAT, SLP-76, which in turn activate MAPK, PLCy signaling pathways. Crystallization studies of the Syk catalytic domain (360-635) showed more activity compared to the full length Syk enzyme.
  • This in vitro assay tests the ability of syk to phosphorylate a substrate peptide in the presence of ATP. By using a radio-labeled form of ATP, it is possible to measure the amount of phosphorylation of the substrate.
  • the enzyme transfers a radio-labeled phosphate group from ⁇ 32 P labeled ATP to pG4T. Briefly the enzyme was incubated with substrate, radio-labeled
  • the reaction mixture was transferred on to a Multiscreen filter plate and unreacted ⁇ 32 P ATP was washed off.
  • the filter plate was dried and the radioactivity was measured on a scintillation counter to estimate the incorporated radioactivity on the substrate. The percent inhibition of activity of the enzyme was calculated by comparing counts in the presence and absence of compounds.
  • the fragment utilized included a C'-terminal tag of 4 amino acids and a stretch of 15 amino acids N-terminal to the kinase domain, starting at amino acid 356.
  • the assay squares were washed 3 times for 5 minutes each in ortho-phosphoric acid (0.5%) and once in acetone. Assay squares were dried for 15 minutes in a 30°C oven and transferred to 96 well optiplate. Microscint-O® reagent (100 ⁇ L, Perkin Elmer) was added to each well, the plate was sealed with Topseal®-A microplates and incubated for 10 minutes at room temperature at very low speed on rocker and the plate was read in the Topcount® NXL instrument.
  • Fold induction radioactivity counts (uncorrected values) in positive control/substrate control.
  • TR-FRET Time-resolved fluorescence resonance energy transfer
  • JAK (Janus kinase 2) is a family of intracellular non-receptor tyrosine kinases that transduce cytokine-mediated signals via the JAK-STAT pathway. These kinases have apparent molecular weight of about 130 Kda. They were initially named “just another kinase” 1 & 2 (since they were just two of a large number of discoveries in a PCR-based screen of kinases), but were ultimately published as "Janus kinase”. JAKs possess two near-identical phosphate-transferring domains. One domain exhibits the kinase activity while the other negatively regulates the kinase activity of the first. They are crucial signal transducers for a variety of cytokines, growth factors and interferons.
  • TR-FRET assays are homogeneous proximity assays where Eu-labeled antiphosphotyrosine antibody binds to the phosphorylated substrates labeled with Ulight fluorescence acceptor. Eu can transfer energy to Ulight accepter in the complex and the interaction of two dye-labeled binding partners is detected by the energy transfer between a donor and an acceptor dye, and the subsequent light emission by the acceptor dye. The intensity of the light emission is proportional to the level of Ulight peptide phosphorylation.
  • the objective of this assay was to examine by Fluorescence method the effect of compounds on ⁇ -hexosaminidase release during immune complex mediated degranulation in RBL2H3 cells.
  • Protocol A 24 well format
  • RBL2H3 cells were maintained in MEM complete media containing 10% FBS at 70% - 80% confluence in a mammalian cell culture C0 2 incubator with 5% C0 2 at 37°C. 2 x 10 5 cells/well were plated in 1 mL of complete media and incubated for 5 hours for cell attachment. Complete media was replaced with 1 mL of serum free MEM media containing 1.2 ⁇ g/mL of anti-DNP rat IgE as sensitizing agent and further incubated overnight with 5% C0 2 at 37°C. The following day, cells were washed with serum free media and further treated with various concentrations of test compounds (in 0.1% DMSO) for 45 minutes at 37°C and 5% C0 2 .
  • test compounds in 0.1% DMSO
  • % release of ⁇ -Hexosaminidase for the test compound was calculated using the following formula:
  • Protocol B 96 well format
  • RBL2H3 cells were maintained in MEM complete media containing 10% FBS at 70-80% confluence in a mammalian cell culture C0 2 incubator with 5% C0 2 at 37°C.
  • 5 x 10 4 cells/well were plated in 200 uL of complete media containing 0.3 ⁇ g/mL of anti-DNP rat IgE as sensitizing agent for 24 hours at 37°C & 5% C0 2 .
  • the following day, cells were washed twice with PIPES buffer for 10 minutes at 37°C and replenished with serum free MEM media. Cells were treated with various concentrations of test compounds (in 0.5% DMSO) for 15 minutes at 37°C and 5% C0 2 .
  • the cells were further stimulated with 0.1 ⁇ g/mL of DNP-BSA for 45 minutes.
  • the plates were spun for 5 minutes at 2000 rpm and 25 ⁇ L of culture supernatant was then transferred from each assay well into a 96 well black coated plate.
  • Fifth ⁇ L ⁇ -NAG substrate was added and incubated at RT for 30 minutes. After incubation with substrate, 150 ⁇ L of stop solution was added and fluorescence was monitored. (Excitation 355 nm; Emission 460 nm). See, Yamamoto, JPET, 306(3):1174- 1181 (2003) and Taylor, MCB, 15(8): 4149-4157 (1995), which are herein incorporated by reference.
  • Example 343 In vivo assay - Chronic study
  • CIA Collagen Induced Arthritis
  • RA rheumatoid arthritis
  • ell type II collagen
  • CIA exhibits several features of human RA such as severe swelling inflammation of joints, synovial hyperplasia, cartilage destruction and bone erosion.
  • Pathophysiology of CIA consists of T cell component, as evidenced by increased infiltration of T-cells in joint synovium and also, by attenuation of CIA in T-cell deficient mice.
  • CIA development involves B cell component too, as is evidenced by circulating ell antibody in disease animals and also, failure to develop the disease in xid mice/B cell deficient mice/CXCR5 null mice. Recently, a significant role of macrophages has also been suggested in the pathogenesis of CIA as well as human RA. See, Pine, "Inflammation and bone erosion are suppressed in models of rheumatoid arthritis following treatment with a novel Syk inhibitor", Clin. Immunol, 2007, 124(3 ):244-57; Xiong cha, "Suppression of the onset and progression of collagen-induced arthritis in rats by QFGJS, a preparation from an anti-arthritic Chinese herbal formula", J.
  • mice Female Lewis rats (8 per group, 6-8 weeks old) were immunized on day 1 with type II collagen (Immunization grade Bovine type II; Chondrex; Cat #20021) emulsified with Complete Freund's Adjuvant (Sigma; Cat# F5881) at a final concentration of 1.2 mg/mL).
  • type II collagen Immunization grade Bovine type II; Chondrex; Cat #20021
  • Complete Freund's Adjuvant Sigma; Cat# F5881
  • a booster injection of the same type II collagen emulsified with Incomplete Freund's Adjuvant (Sigma, Cat #F5506) (0.25 mL/rat) was given to the animals on day 8 at the base of the tail (100 ⁇ g).
  • the final ell concentration in the booster was 0.4 ⁇ g/mL.
  • Animals with an arthritic score of > 1 were grouped and dosing with test compound (30 mg/kg bid) or methotrexate (0.5 mg/kg) started between about day 12 to day 14, with daily dosing of their respective compounds continuing for 10 days.
  • Edema Paw volumes are measured by Plethysmometry for the animals before induction of CIA (Basal readings) and on Day 1, 3, 6 and 9 of dosing period. Both hind paw volumes were measured and edema was calculated by subtracting from the basal mean.
  • Example 344 In vivo assay - Acute study
  • Type III hypersensitivity reactions are immune complex-mediated, and involve the deposition of antigen/antibody complexes mainly in the vascular walls, serosa (pleura, pericardium, synovium), and glomeruli. This involves formation of antigen antibody complexes after the intradermal injection of an antibody. If the animal was previously injected with antigen and dye (has circulating antigen), an Arthus reaction occurs. This manifests as local vasculitis due to deposition of immune complexes in dermal blood vessels. Activation of complement and recruitment of PMNs ensue, resulting in an inflammatory response and extravasation of dye to the skin. Compounds which can inhibit this complex process can have therapeutic implications in wide range of inflammatory and auto-immune disorders. See, Pine,
  • mice Female c57BL/6 mice were given an antigen injection in which the antigen was 0.1% Ovalbumin (OVA) in PBS containing 1% Evans blue (EB) at the concentration of 10 mL/kg intravenously under Isoflurane anesthesia [2.5 mg/mouse with a body weight of 25 g].
  • Ovalbumin Ovalbumin
  • EB Evans blue
  • PBS phosphate buffered saline
  • mice were euthanized by cervical dislocation 4 hours after antigen (Ovalbumin) challenge. Skin tissue was assessed for edema by tracing the edema area on to a transparent plastic sheet. Punch biopsies of the injection sites were collected.
  • Edema area was measured manually by scale. Two diameters were taken and averaged for each animal.
  • This example illustrates that the compounds described herein may be utilized in treating inflammation.
  • combination treatments for treating cancers involving targeted oncologic therapies e.g., inhibition of specific biochemical pathways
  • immunotherapeutic agents would be expected to decrease treatment efficacy and/or increase adverse side effects, because targeted therapies with immuno stimulatory potential can also display immunosuppressive activities.
  • the activity of targeted therapies may therefore cancel out, and even inhibit, therapeutic mechanisms of imunomodulators.
  • the combination of dual inhibitors of Syk/JAK family kinases according to the disclosure and immunotherapeutic agents for the treatment of cancers would be expected to result in a decrease in efficacy.
  • such combination treatments can result in increases in toxicity and adverse side effects, for example by increasing cytokine production and other inflammatory reactions related to the non-tumor suppressive activities of the immunotherapeutic agents.
  • compositions comprising a compound of Formula (I), as described herein, and one or more immunotherapeutic agents can be used to treat cancers, where the combination has a reduced adverse side effect profile and where the activity of the immunotherapeutic agent is not suppressed.
  • the anticancer or antitumor activity of pharmaceutical combinations of the present disclosure may be substantially the same as, or may be increased, as compared to administering a compound of Formula (I) alone.
  • the pharmaceutical combinations of the present disclosure can reduce negative side effects in patients, for example by inhibiting or suppressing chemokine and/or cytokine levels.
  • a pharmaceutical combination comprising the combination comprising a compound of Formula (I), as described herein, and at least one immunotherapeutic agent.
  • the at least one immunotherapeutic agent can comprise one or more immunomodulators.
  • the immunomodulators can target one or more components of the immune system, including (without limitation) PD-1, PD-L1, CTLA-4, 4-1BB, OX40, LAG3, GITR, TIM3, VISTA, KIR, ICOS, BTLA, CD244, CD80, CD86, PD-L2, IDO-1, IDO-2, and B7-H3, and can produce a therapeutic effect.
  • the one or more immunomodulators can comprise (without limitation) inhibitors of immune system components, activators of immune system components, and/or immune checkpoint inhibitors, and can comprise biologic or small molecule therapeutics.
  • TCR agents can include (without limitation) chimeric antigen receptor (CAR) T cells, and TCR agonist or antagonist peptides.
  • CAR T cells can comprise (without limitation) CTL019 and JCAR015 etc.
  • the vaccines can include (without limitation) sipuleucel-T (Provenge ® ) and Talimogene laherparepvec (Imlygic ® ), aslo known as T-Vec.
  • checkpoint inhibitors can include (without limitation) co- inhibitory molecules such as cytotoxic T-lymphocyte-as so dated protein 4 (CTLA-4), PD-1, lymphocyte-activation gene 3, and T-cell immunoglobulin mucin-3, and co-stimulatory molecules such as glucocorticoid-induced tumor necrosis factor receptor and OX40 (CD134, TNFRSF4, tumor necrosis factor receptor superfamily member 4).
  • co- inhibitory molecules such as cytotoxic T-lymphocyte-as so dated protein 4 (CTLA-4), PD-1, lymphocyte-activation gene 3, and T-cell immunoglobulin mucin-3
  • co-stimulatory molecules such as glucocorticoid-induced tumor necrosis factor receptor and OX40 (CD134, TNFRSF4, tumor necrosis factor receptor superfamily member 4).
  • the immune checkpoint inhibitors can include (without limitation), anti-PD-1 antibody, anti-PD- Ll antibody, anti-CTLA-4 antibody, anti-4-lBB antibody, anti-OX40 antibody, anti-LAG3 antibody, anti-GITR antibody, anti-TIM3 antibody, anti- VISTA antibody, anti-KIR antibody, anti-ICOS antibody, anti-BTLA antibody, anti-CD244 antibody, anti-CD80 antibody, anti- CD86 antibody, anti-PD-L2 antibody, anti-IDO-1 antibody, anti-IDO-2 antibody, anti-B7-H3 antibody, and small molecule inhibitors of any of these antibody targets, for example an IDO inhibitor drug.
  • the immune checkpoint inhibitor is anti-PD-1 antibody.
  • the immune checkpoint inhibitor is small molecule IDO-1 inhibitor.
  • the pharmaceutical combination can further comprise at least one pharmaceutically acceptable carrier.
  • the disclosure also provides methods of treating cancer, the methods comprising administering to a patient in need thereof a multi-part pharmaceutical combination comprising a compound of Formula (I), as described herein, and at least one immunotherapeutic agent, as described herein.
  • the compounds of Formula (I) can be administered intravenously or orally, or by any other suitable manner.
  • the immunotherapeutic agent can be administered intravenously, subcutaneously, intramuscularly, or orally, or by any other suitable manner.
  • administration of the compound is intravenous and the administration of the at least one immunotherapeutic agent is intravenous, subcutaneous or intra-muscular.
  • administration of the compound is oral and the administration of the at least one immunotherapeutic agent is oral.
  • administration of the compound is intravenous and the administration of the at least one immunotherapeutic agent is oral. In yet another embodiment, administration of the compound is oral and the administration of the at least one immunotherapeutic agent is intravenous, subcutaneous or intra-muscular.
  • the compound of Formula (I) and the immunotherapeutic agent can be administered by any suitable dosing regimen.
  • the compound of Formula (I) is administered once or twice daily.
  • the compound and the at least one immunotherapeutic agent can be administered simultaneously or sequentially.
  • the compound and the at least one immunotherapeutic agent are administered sequentially.
  • the at least one immunotherapeutic agent is administered intermittently every four to thirty days.
  • the compound of Formula (I) can be administered to a patient at a dosage of about 5 mg/kg to about 70 mg/kg, preferably from about 10 mg/kg to about 60 mg kg. In an embodiment, the compound of Formula (I) is administered at a dosage of about 10 mg/kg, about 30 mg/kg, or about 60 mg/kg. In other embodiments, the compound of Formula (I) can be administered to a patient at a dosage of about 10 mg, 20 mg, 30 mg, 40 mg, 40 m g, 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 90 mg, 100 mg or 120 mg once or twice per day.
  • cancers can be treated using the pharmaceutical combinations provided here, including (without limitation) the following cancers: prostate, head, neck, eye, mouth, throat, esophagus, bronchus, larynx, pharynx, chest, bone, lung, colon, rectum, stomach, bladder, uterus, cervix, breast, ovaries, vagina, testicles, skin, thyroid, blood, lymph nodes, kidney, liver, intestines, pancreas, brain, central nervous system, adrenal gland, skin or a leukemia and/or lymphoma.
  • methods of the present disclosure include ⁇ identifying an indication or patient population for administering the pharmaceutical combinations of the present disclosure.
  • the disclosure provides methods of inhibiting tumor growth or metastasis in a subject, the methods comprising administering to the subject a multi-part pharmaceutical combination comprising a compound of Formula (I), as described herein, and at least one immunotherapeutic agent, as described herein, wherein the administration of the pharmaceutical combination inhibits tumor growth or metastasis.
  • Inhibition of tumor growth or metastasis means reducing the number of cancer cells or causing the amount of cancer cells to remain substantially the same, reducing tumor size or causing the tumor size to remain substantially the same, inhibiting metastasis (including inhibition of tumor cell migration and/or invasion), inhibiting tumor growth and/or ameliorating one or more of the symptoms of the cancer.
  • TGI tumor growth or metastasis
  • a therapeutically effective amount of a pharmaceutical combination when used for the treatment of cancer is an amount which may inhibit tumor growth or metastasis.
  • administration of the pharmaceutical combinations described herein can achieve TGI in an amount of 0% to 100%, preferably an amount of above about 50%, for example about 50% to 90%, or about 60% to 80%.
  • the disclosure provides dosing regimens comprising administering to a patient in need a multi-part pharmaceutical combination comprising a compound of Formula (I), as described herein, and at least one immunotherapeutic agent, as described herein. Administration of the compound of Formula (I) and the immunotherapeutic agent can be performed as described herein.
  • kits comprising at least one first dosage form comprising a compound of Formula (I), as described herein, and at least one second dosage form comprising at least one immunotherapeutic agent, as described herein, and administering to a patient in need the compound of Formula (I) and the immunotherapeutic agent.
  • Administration of the compound of Formula (I) and the immunotherapeutic agent can be performed as described herein.
  • the disclosure provides uses of a medicament in treating cancer, wherein the medicament is administered by a dosing regimen comprising administering to a patient in need a compound of Formula (I), as described herein, and at least one immunotherapeutic agent, as described herein.
  • Administration and the dosing regimen of the compound of Formula (I) and the immunotherapeutic agent can be performed as described herein.
  • Example 345 demonstrates that the pharmaceutical combinations described herein can suppress cytokines and can prevent cytokine release syndrome. Suppression of cytokines can result in a decrease in adverse side effects and toxicity related to cytokine production in patients.
  • EXAMPLE 345 Evaluation of inflammation biomarkers in cancer patients treated with COMPOUND A
  • Example 189 2-(l-(4-((4-(4-hydroxypiperidin-l-yl)phenyl)amino)-5-oxo-5,6- dihydropyimido[4,5-d]pyridazin-2-yl)piperidm-4-yl)acetonitrile, was tested as described below.
  • the compound of Example 189 is sometimes referred to herein as "COMPOUND A.”
  • Serum samples were collected from patients before the treatment with COMPOUND A on Day 1 and after treatment with COMPOUND A on day 15. The samples were then analyzed for the levels of a panel of 45 inflammation biomarkers. The percent change in the serum levels of each biomarker was calculated by comparing the protein levels before and after treatment. There was a significant inhibition of several inflammation biomarkers on Day 15 at all doses evaluated. A few key inflammation biomarkers that showed a marked inhibition with COMPOUND A treatment were C-Reactive Protein (CRP), p2-microglobulin
  • C-reactive protein is an acute-phase protein, the levels which are elevated in response to inflammation. The levels of this protein are also elevated in patients with various types of cancers and this is considered as a potential predictor of cancer risk and/or survival.
  • p2-microglobulin is a component of MHC class I molecules and is encoded by the gene, B2M. ⁇ 2 -microglobulin is found to be elevated in lymphoma and multiple myeloma.
  • IL-18 is a proinflammatory cytokine involved in several inflammatory disorders.
  • ⁇ - ⁇ also known as CCL4
  • CCR5 chemokine receptor 5
  • the levels of all these inflammation biomarkers have been significantly inhibited in patient serum after treatement with COMPOUND A. Because of its ability to suppress some of the important cytokines such as IL-18, COMPOUND A also has a potential to prevent the cytokine release syndrome when combined with immunotherapies (see also Fig. 3).
  • COMPOUND A was also evaluated for its ability to inhibit signaling pathways (JAK/STAT) in peripheral blood monocular cells stimulated with varuious cytokines such as IL-2, IL-4, IL-6, IL-12, IL-23 etc. In these studies, COMPOUND A potently inhibited the
  • HPC Hydroxypropylcellulose LOQ: Limit of quantification
  • TGI Tumor growth inhibition
  • IDO-1 Indoleamine 2,3-dioxygenase
  • PD-1 Programmed cell Death- 1
  • the inventors have unexpectedly discovered that administration of the pharmaceutical combinations comprising a compound of Formula (I) and an immunotherapeutic agent, as described herein, can produce substantially the same, or an increase in, anti-tumor efficacy compared to administering the compound without an immunotherapeutic agent.
  • Example 346 demonstrates the anti-tumor efficacy of COMPOUND A alone and in combination with anti-PDl antibody or IDO-1 inhibitor (epacadostat) in a 4T1 - Balb/c mouse syngeneic allograft tumor model.
  • the mice bearing 4T1 allografts in the untreated control group showed a physiological body weight gain of 19 %.
  • COMPOUND A, anti PD-1 antibody, epacadostat, or their combinations were tolerated well with a minimal, yet acceptable, loss of body weight (-2 %).
  • Plasma COMPOUND A concentrations were found to be in the range of 8 to 1813 ng ml; however in tumor samples, it ranged from 72 to 2495 ng/g for mice administered with 10 to 60 mg/kg
  • COMPOUND A There was an 8-fold increase in exposure in tumor vs. plasma samples for single agent COMPOUND A at 10 mg/kg; and a 3-fold increase was obtained when COMPOUND A at 10 mg/kg combined with either anti PD-1 antibody or epacadostat. A lower COMPOUND A concentration in tumor vs. plasma was found at 30 - 60 mg/kg COMPOUND A when combined with anti PD1 antibody. Similarly, COMPOUND A-60 mg kg treatment combined with epacadostat resulted in lower concentration of COMPOUND A in tumor than plasma. No treatment related morbidity or mortality was seen during the study.
  • Epacadostat an inhibitor of IDO-1 has shown efficacy in inhibiting tumor growth in multiple immunocompetant tumor models.
  • PD-1 Programmed cell Death- 1 expression by tumor- infiltrating lymphocytes is shown to be important in anti-inflammatory and tissue injury protection that further mediates tumor-induced immune suppression/immune escape. Since tumor growth inhibitory effect of both anti-PDl antibody and epacadostat have already been established in this model, a single dose of these agents was used in combination with different doses of COMPOUND A to demonstrate the potential benefit to tumor growth inhibition demonstrated by COMPOUND A.
  • mice obtained from Envigo, Netherlands.
  • the animals were housed in individually ventilated cages (maximum of 5 animals/cage) with 12 hour dark, 12 hour light conditions.
  • the animals were fed food and water ad libitum.
  • Temperature and relative humidity were maintained at 20 ⁇ 2°C and 65%, respectively.
  • COMPOUND A (Lot # C15K081007A) was formulated in the vehicle containing 20 mg ml hydroxypropylcellulose (HPC, Klucel LF). The formulation was stirred for 30 min to obtain a homogenous suspension to which polysorbate-80 was added at 1 mg/ml. This formulation was continuously stirred throughout dosing.
  • Epacadostat (Cat # PBLJ9203; CAS # 1204669-58-8, purchased from PharmaBlock): was formulated as a homogenous suspension in the vehicle containing 0.5 % methyl cellulose (MC). The formulation was stirred for 30 min to obtain a homogenous suspension to which polysorbate-80 was added at 1 mg/ml. This formulation was continuously stirred throughout the dosing.
  • Anti-mouse PD1 antibody 100 ⁇ g in 200 ⁇ antibody was administered (i.p.) per mouse.
  • Antibody was diluted with 1 X sterile PBS (calcium and magnesium-free).
  • COMPOUND A and epacadostat were administered orally by gavage at a dose volume of 10 mL/kg.
  • Mouse breast carcinoma 4T1 cells were sourced from American Type Culture Collection (ATCC), USA. Cells were grown in RPMI-1640 medium (Sigma, Cat # R6504) supplemented with 10% FBS (Invitrogen, Cat # 10438-026), and 1% penicillin streptomycin (Thermo Fisher Scientific, Cat # 15140-122). To establish allografts, 1 million 4T1 cells were suspended in 50 ⁇ of serum-free medium and mixed at 1:1 ratio with matrigel before implanting subcutaneously into the right flank of mice.
  • ATCC American Type Culture Collection
  • Tumor dimensions were measured for all animals on the first treatment day (Day 1) and then two times per week, including the termination day of the study. Tumor volumes were calculated using the formula: tumor length x (tumor width) 2 / 2. Tumor growth inhibition was calculated after normalizing the tumor volume on a given day to that on day 1.
  • Treatment was initiated 9 days after subcutaneous injection of tumor cells when the average tumor volume reached approximately 90 mm 3 .
  • the animals were randomized based on tumor volumes into 12 groups of eight animals each.
  • the dosing schedule of COMPOUND A and epacadostat were twice daily for 15 days by p.o. and anti-PDl antibody was administered once in 4 days (total 5 doses) by i.p.
  • the treatment period was 15 days after which the overall efficacy and tolerability was evaluated based on tumor volume and body weight changes observed during the treatment period.
  • Tumor volumes were analyzed using two-way ANOVA with Bonferonni' s multiple comparisons test for comparison of treatment versus control group.
  • %TGI percent tumor growth inhibition
  • the anti-tumor effect of COMPOUND A was evaluated in tumor allograft bearing Balb/c mice.
  • the anti-tumor efficacy of single agent anti-PDl antibody, epacadostat and COMPOUND A were compared to vehicle control, whereas combination groups, COMPOUND A + anti-PDl antibody, and COMPOUND A + epadacostat were compared with single agent groups and vehicle control.
  • mice treated with COMPOUND A (10, 30, 60 mg kg), anti-PDl antibody with untreated vehicle control indicated a statistically significant decrease ( ⁇ .0001) on day 15 of treatment.
  • the average tumor volume for mice in vehicle control group was 976 ⁇ 86 mm 3 and that for 10, 30, 60 mg/kg COMPOUND A, anti-PDl antibody 0.1 mg/mouse, and epacadostat 30 mg kg treated groups were found to be 549 ⁇ 51 (48 % TGI), 447 ⁇ 38 (60 % TGI), 282 ⁇ 66 (79 % TGI), 530 ⁇ 58 (51 % TGI), and 567 ⁇ 48 mm 3 (46 % TGI), respectively ( Figure 4 and 5; Table 18).
  • mice bearing 4T1 allografts in the untreated control group showed a physiological body weight gain of 19 % (Table 20).
  • Mice treated with COMPOUND A - 10, 30 and 60 mg/kg equally gained 17 % body weight on day 15, not significantly different from controls (p>0.05, Table 19; Figures 6-9).
  • Anti-PDl antibody combinations with COMPOUND A - 10, 30, and 60 mg kg had a similar gain of 18 %, 17 %, and 16 % body weights, respectively ( Figures 6 and 8; Tables 19 and 20).
  • Table 19 Average body weight in mice after different days of treatment initiation
  • COMPOUND A anti PD-1 antibody, and epacadostat were tolerated well with a minimal, yet acceptable, loss of body weight. No treatment related morbidity or mortality was seen during the study. Plasma and tumor concentrations of COMPOUND A
  • Plasma and tumor samples from six mice/group were collected for analyzing COMPOUND A by LC-MS.
  • Plasma COMPOUND A concentrations were found to be in the range of 8 to 1813 ng/ml (Table 21); however in tumor samples, COMPOUND A concentrations ranged from 72 to 2495 ng/g for mice administered with 10 to 60 mg/kg COMPOUND A (Table 22).
  • concentration of COMPOUND A in plasma ranged from 41 to 1889 ng/ml and in tumor it ranged from 126 to 1624 ng/g (Tables 21, 22).
  • concentration of COMPOUND A in plasma ranged from 48 to 4114 ng/ml and in tumor it ranged from 172 to 2517 ng/g in mice administered with 10 to 60 mg/kg COMPOUND A (Tables 21, 22).
  • COMPOUND A-10 mg/kg There was an 8-folds increase in exposure in tumor than plasma samples for single agent COMPOUND A-10 mg/kg; and a 3-fold increase was obtained when COMPOUND A-10 mg/kg combined with either anti PD-1 antibody or epacadostat.
  • a lower COMPOUND A concentration in tumor than plasma was found at 30 - 60 mg/kg COMPOUND A when combined with anti PD 1 antibody.
  • COMPOUND A-60 mg kg treatment combined with epacadostat resulted in lower concentration of COMPOUND A in tumor than plasma.
  • COMPOUND A at 10, 30 and 60 mg/kg doses resulted in a significant and dose- dependent tumor growth inhibition of 48 %, 60 %, and 79 %.
  • a tumor growth inhibition of 51 % and 46 % was obtained for anti-PDl antibody (0.1 mg/mouse) and epacadostat (30 mg kg), respectively. All treatments were tolerated well and no treatment related morbidity or mortality was seen during the study.
  • Combination of COMPOUND A 10 mg kg with anti- PDl antibody showed substantially the same TGI as that of single agent COMPOUND A.
  • There was an enhancement in the tumor growth inhibition when epacadostat combined with COMPOUND A although it was not statistically significant (p>0.05).
  • COMPOUND A- 10 mg/kg A 3 -folds increase was also obtained when COMPOUND A- 10 mg/kg combined with either anti PD-1 antibody or epacadostat.
  • Example 347 demonstrates the anti-tumor efficacy of COMPOUND A alone and in combination with anti-PDl antibody or IDO-1 inhibitor (epacadostat) in B16-F10 - C57/BL6 syngeneic allograft tumor model.
  • the objective of this study was to test efficacy of COMPOUND A in combination with anti-PDl antibody or IDO-1 inhibitor (epacadostat) using B16-F10 - C57/BL6 syngeneic allograft tumor model.
  • the average tumor volume for mice in vehicle control group was 2529 ⁇ 110 mm 3 and that for 10, 20, 60 mg/kg COMPOUND A, anti-PDl antibody 0.1 mg/mouse, and epacadostat 30 mg/kg treated groups were found to be 1323 ⁇ 121 (49 % TGI), 1041 ⁇ 98 (60 % TGI), 923 ⁇ 58 (65 % TGI), 1117 ⁇ 82 (57 % TGI), and 1394 ⁇ 90 mm 3 (46 % TGI), respectively.
  • Co-administration of either anti-PDl antibody or epacadostat with COMPOUND A showed substantially the same tumor growth reduction in B16-F10 melanoma cancer syngeneic model.
  • COMPOUND A (10 - 60 mg/kg), anti-PDl antibody and epacadostat were tolerated well without loss of body weight.
  • Combinations of COMPOUND A with anti-PDl antibody or epacadostat were tolerated well without loss of body weight.
  • a widely used B16-F10 tumor was employed in C57/BL6 mouse model to assess the immunomodulatory effect of known immunotherapeutic agents, anti-PDl antibody and IDO- 1 inhibitor (epacadostat) in combination with COMPOUND A. Since tumor growth inhibitory effect of both anti-PDl antibody and epacadostat have already been established in this model, a dose that has resulted in 40-50% inhibition was selected as a single agent and combined with different doses of COMPOUND A to understand any potential benefit to tumor growth inhibition demonstrated by COMPOUND A.
  • the animals were housed in individually ventilated cages (maximum of 5 animals/cage) with 12 hour dark, 12 hour light conditions. The animals were fed food and water ad libitum. Temperature and relative humidity were maintained at 20 ⁇ 2°C and 65%, respectively.
  • COMPOUND A (Lot # C15K081007A) was formulated in the vehicle containing 20 mg/ml hydroxypropylcellulose (HPC, Klucel LF). The formulation was stirred for 30 min to obtain a homogenous suspension to which polysorbate-80 was added at 1 mg/ml. This formulation was continuously stirred throughout dosing.
  • Epacadostat (Cat # PBLJ9203; CAS # 1204669-58-8, purchased from PharmaBlock): was formulated as a homogenous suspension in the vehicle containing 0.5 % methyl cellulose (MC). The formulation was stirred for 30 min to obtain a homogenous suspension to which polysorbate-80 was added at 1 mg/ml. This formulation was continuously stirred throughout the dosing.
  • Anti-mouse PD1 antibody 100 ⁇ g in 200 ⁇ antibody was administered (i.p.) per mouse. Antibody was diluted with 1 X sterile PBS (calcium and magnesium-free). COMPOUND A and epacadostat were administered orally by gavage at a dose volume of 10 mL/kg.
  • Cell line and tumor model 100 ⁇ g in 200 ⁇ antibody was administered (i.p.) per mouse. Antibody was diluted with 1 X sterile PBS (calcium and magnesium-free). COMPOUND A and epacadostat were administered orally by gavage at a dose volume of 10 mL/kg.
  • Cell line and tumor model 100 ⁇ g in 200 ⁇ antibody was administered (i.p.) per mouse. Antibody was diluted with 1 X sterile PBS (calcium and magnesium-free). COMPOUND A and epacadostat were administered orally by gavage at a dose volume of 10 mL/kg.
  • Cell line and tumor model 100 ⁇ g in
  • Mouse melanoma B16-F10 cells were sourced from American Type Culture Collection (ATCC), USA. Cells were grown in Dulbecco's Modified Eagel Medium (DMEM) (Gibco, Cat # 31600-034) supplemented with 10% FBS (Invitrogen, Cat # 10438- 026), and 1% penicillin streptomycin (Thermo Fisher Scientific, Cat # 15140-122). To establish allografts, 0.1 million B16-F10 mouse melanoma cells were suspended in 50 ⁇ of serum-free medium and mixed at 1 : 1 ratio with matrigel before implanting subcutaneously into the right flank of mice.
  • DMEM Dulbecco's Modified Eagel Medium
  • FBS Invitrogen, Cat # 10438- 026
  • penicillin streptomycin Thermo Fisher Scientific, Cat # 15140-122
  • Tumor dimensions were measured for all animals on the first treatment day (Day 1) and then two times per week, including the termination day of the study. Tumor volumes were calculated using the formula: tumor length x (tumor width) 2 / 2. Tumor growth inhibition was calculated after normalizing the tumor volume on a given day to that on day 1.
  • Drug Treatments Treatment was initiated 6 days after subcutaneous injection of tumor cells when the average tumor volume had reached approximately 50 mm 3 .
  • the animals were randomized based on tumor volumes into 12 groups of ten animals each.
  • the dosing schedule of COMPOUND A and epacadostat were twice daily for 15 days by p.o. and anti-PDl antibody was administered once in 4 days (total 5 doses) by i.p.
  • the treatment period was 15 days after which the overall efficacy and tolerability was evaluated based on tumor volume and body weight changes observed during the treatment period.
  • Tumor volumes were analyzed using two-way ANOVA with Bonferonni' s multiple comparisons test for comparison of treatment versus control group.
  • %TGI percent tumor growth inhibition
  • % TGI [1 -(Treatment TVpinai - Treatment TV ln i t i_i) / (Control TVpinai - Control
  • the anti-tumor effect of COMPOUND A was evaluated in tumor allograft bearing C57/BL6 mice.
  • the anti-tumor efficacy of single agents, anti-PDl antibody, epacadostat and COMPOUND A were compared to vehicle control, whereas combination groups, COMPOUND A + anti-PDl antibody, and COMPOUND A + epadacostat were compared with single agent groups and vehicle control.
  • mice in vehicle control group was 2529 ⁇ 110 mm 3 and that for 10, 20, 60 mg/kg COMPOUND A, anti-PDl antibody 0.1 mg/mouse, and epacadostat 30 mg/kg treated groups were found to be, respectively 1323 ⁇ 121 (49 % TGI), 1041 ⁇ 98 (60 % TGI), 923 ⁇ 58 (65 % TGI), 1117 ⁇ 82 (57 % TGI), and 1394 ⁇ 90 mm 3 (46 % TGI) ( Figure 17 and 18; Table 26).
  • Co-administration of anti-PDl antibody showed substantially the same tumor growth inhibition obtained for COMPOUND A in B16-F10 tumor syngeneic model (53 %, 62% and 74% TGI for combinations of anti PD-1 antibody and COMPOUND A vs. 49 %, 60% and 65% TGI for single agent COMPOUND A at 10, 20 and 60 mg kg, respectively) ( Figure 17 and 18; Table 26).
  • there was substantially the same benefit of coadministration of epacadostat with COMPOUND A 52 %, 64% and 70% TGI for combinations of epacadostat and COMPOUND A vs. 49 %, 60% and 65% TGI for single agent COMPOUND A at 10, 30 and 60 mg kg, respectively; Figure 17 and 18; Table 26).
  • Table 26 Average tumor volume (mm ) in mice after different days of treatment initiation
  • mice bearing B16-F10 allografts in the untreated control group maintained body weight with negligible change of -0.6 % on day 15 (Table 28).
  • COMPOUND A - 10, 30, and 60 mg/kg, anti-PDl antibody and epacadostat were tolerated well without loss of body weight ( Figures 19 and 21 ; Tables 27 and 28).
  • the highest increase of 17.3 % in body weight was observed in mice treated with mouse anti-PDl antibody (group 5).
  • Table 27 Average body weight of mice after different days of treatment initiation
  • Table 29 Animal mortality was random across all experimental groups and the mortality did not appear to be due to treatment, but could be because of spontaneous metastatic nature of B16F10 cells in the syngeneic C57-BL6 mouse model. Table 29: Animal mortality after different days of treatment initiation
  • Plasma COMPOUND A concentrations were found to be in the range of 32.8 to 935 ng/ml (Table 30), however in tumor samples, COMPOUND A concentrations increased from 883 to 7568 ng/g for mice administered with 10 to 60 mg/kg COMPOUND A (Table 31).
  • concentration of COMPOUND A in plasma ranged from 25 to 1023 ng/ml (Table 30) and in tumor it ranged from 1024 to 9439 ng/g (Table 31).
  • CONCLUSION COMPOUND A at 10, 30 and 60 mg/kg doses resulted in a significant and dose- dependent tumor growth inhibition of 49, 60 and 65%.
  • a tumor growth inhibition of 57 % and 46 % was obtained for anti-PDl antibody (0.1 mg/mouse) and epacadostat (30 mg/kg), respectively.
  • Combination of either anti-PDl antibody or epacadostat showed substantially the same TGI than that obtained from single agent COMPOUND A.
  • Example 348 demonstrates the anti-tumor efficacy of COMPOUND A alone and in combination with anti-PDl antibody or IDO-1 inhibitor (epacadostat) in CT26 - Balb/c syngeneic allograft tumor model
  • the objective of this study was to demonstrate the anti-tumor acitivity of COMPOUND A in combination with anti-PDl antibody or IDO-1 inhibitor (epacadostat) using CT26 - Balb/c syngeneic allograft tumor model.
  • the mice bearing CT26 allografts in the untreated control group showed a physiological body weight gain of 18 %.
  • COMPOUND A - 30 and 60 mg/kg treatment has significantly decreased the physiological gain body weight on day 15 compared to controls (p ⁇ 0.05), however there was no loss of body weight compared to their day 1 initial weights (p ⁇ 0.05).
  • Combination of COMPOUND A with anti-PDl antibody or epacadostat had a significant but acceptable loss of body weight compared to controls (p ⁇ 0.0001).
  • Anti PD-1 antibody or epacadostat co-administration decreased the COMPOUND A concentrations both in plasma and tumor. There was a 2-folds increase in exposure in tumor than plasma samples for single agent COMPOUND A-10 mg/kg, a similar increase was also obtained when combined with either anti PD-1 antibody or epacadostat.COMPOUND A, anti PD-1 antibody, and epacadostat were tolerated with acceptable loss of body weight. An increase in the loss of body weight, still within an acceptable limit of 10 % was observed when combined with anti PD-1 antibody or epacadostat.
  • CT26-Balb/c model was used to assess the immunomodulatory effect of known immunomodulatory agents, anti-PDl antibody and IDO-1 inhibitor (epacadostat) in combination with COMPOUND A. Since tumor growth inhibitory effect of both anti-PDl antibody and epacadostat have already been established in this model, a single dose of these agents was used in combination with different doses of COMPOUND A to understand any potential benefit to tumor growth inhibition demonstrated by COMPOUND A.
  • the animals were housed in individually ventilated cages (maximum of 5 animals/cage) with 12 hour dark, 12 hour light conditions. The animals were fed food and water ad libitum. Temperature and relative humidity were maintained at 20 ⁇ 2°C and 65%, respectively.
  • COMPOUND A (Lot # C15K081007A) was formulated in the vehicle containing 20 mg ml hydroxypropylcellulose (HPC, Klucel LF). The formulation was stirred for 30 min to obtain a homogenous suspension to which polysorbate-80 was added at 1 mg/ml. This formulation was continuously stirred throughout dosing.
  • Epacadostat (Cat # PBLJ9203; CAS # 1204669-58-8, purchased from PharmaBlock): was formulated as a homogenous suspension in the vehicle containing 0.5 % methyl cellulose (MC). The formulation was stirred for 30 min to obtain a homogenous suspension to which polysorbate-80 was added at 1 mg/ml. This formulation was continuously stirred throughout the dosing.
  • Anti-mouse PD1 antibody 100 ⁇ g in 200 ⁇ antibody was administered (i.p.) per mouse.
  • Antibody was diluted with 1 X sterile PBS (calcium and magnesium-free).
  • COMPOUND A and epacadostat were administered orally by gavage at a dose volume of 10 mL/kg.
  • Mouse colon carcinoma CT26 cells were sourced from American Type Culture Collection (ATCC), USA. Cells were grown in RPMI-1640 medium (Sigma, Cat # R6504) supplemented with 10% FBS (Invitrogen, Cat # 10438-026), and 1% penicillin streptomycin (Thermo Fisher Scientific, Cat # 15140-122). To establish allografts, 1 million CT26 cells were suspended in 50 ⁇ of serum-free medium and mixed at 1:1 ratio with matrigel before implanting subcutaneously into the right flank of mice.
  • ATCC American Type Culture Collection
  • Tumor dimensions were measured for all animals on the first treatment day (Day 1) and then two times per week, including the termination day of the study. Tumor volumes were calculated using the formula: tumor length x (tumor width) 1 12. Tumor growth inhibition was calculated after normalizing the tumor volume on a given day to that on day 1.
  • Treatment was initiated 9 days after subcutaneous injection of tumor cells when the average tumor volume reached approximately 90 mm 3 .
  • the animals were randomized based on tumor volumes into 12 groups of eight animals each.
  • the dosing schedule of COMPOUND A and epacadostat were twice daily for 15 days by p.o. and anti-PDl antibody was administered once in 4 days (total 5 doses) by i.p.
  • the treatment period was 15 days after which the overall efficacy and tolerability was evaluated based on tumor volume and body weight changes observed during the treatment period.
  • Tumor volumes were analyzed using two-way ANOVA with Bonferonni' s multiple comparisons test for comparison of treatment versus control group.
  • %TGI percent tumor growth inhibition
  • % TGI [1 -(Treatment TVFinai - Treatment TV ln iti_i) / (Control TVFinai - Control
  • the average tumor volume for mice in vehicle control group was 2835 ⁇ 500 mm 3 and that for 10, 30, 60 mg/kg COMPOUND A, anti-PDl antibody 0.1 mg/mouse, and epacadostat 30 mg/kg treated groups were found to be 1929 ⁇ 407 (33 % TGI), 965 ⁇ 135 (68 % TGI), 771 ⁇ 90 (76 % TGI), 1264 ⁇ 143 (57 % TGI), and 1901 ⁇ 396 mm 3 (34 % TGI), respectively ( Figure 30 and 31 ; Table 35).
  • Co-administration of COMPOUND A- 10 mg/kg and anti-PDl antibody in CT26 tumor syngeneic model significantly increased tumor growth inhibition (p ⁇ 0.0 ⁇ , 63 % TGI in combination vs.
  • Table 35 Average tumor volume (mm 3 ) in mice after different days of treatment initiation
  • Table 36 Average body weight in mice after different days of treatment initiation
  • COMPOUND A anti PD-1 antibody, and epacadostat were tolerated with acceptable loss of body weight. An increase in the loss of body weight, still within an acceptable limit of 10 % was observed when combined with anti PD-1 antibody or epacadostat.
  • Plasma and tumor samples from six mice/group were collected for analyzing COMPOUND A by LC-MS.
  • Plasma COMPOUND A concentrations were found to be in the range of 208 to 3071 ng/ml (Table 38), however in tumor samples, COMPOUND A concentrations ranged from 382 to 1286 ng/g for mice administered with 10 to 60 mg/kg COMPOUND A (Table 39).
  • concentration of COMPOUND A in plasma ranged from 80 to 711 ng/ml and in tumor it ranged from 147 to 805 ng/g (Tables 38, 39).
  • concentration of COMPOUND A in plasma ranged from 51 to 1774 ng/ml and in tumor it ranged from 151 to 937 ng/g in mice administered with 10 to 60 mg/kg COMPOUND A (Tables 38, 39).
  • COMPOUND A- 10 mg/kg There was a 2-fold increase in exposure in tumor than plasma samples for single agent COMPOUND A- 10 mg/kg, a similar increase was also obtained when combined with either anti PD-1 antibody or epacadostat.
  • COMPOUND A at 10, 30 and 60 mg/kg doses resulted in a significant and dose- dependent tumor growth inhibition of 33, 68 and 79%.
  • a tumor growth inhibition of 57 % and 34 % was obtained for anti-PDl antibody (0.1 mg/mouse) and epacadostat (30 mg/kg), respectively.
  • Combination of COMPOUND A 10 mg/kg with anti-PDl antibody significantly enhanced TGI of single agent COMPOUND A; however, higher concentrations of COMPOUND A did not produce this response.
  • TGI was substantially the same for COMPOUND A and epacadostat combination compared to single agent COMPOUND A.
  • Example 349 demonstrates the clinical activity, safety and tolerabilityof
  • COMPOUND A a dual SYK/JAK inhibitor, in patients with Non-Hodgkins lymphoma (NHL) and solid tumors, including dose-limiting toxicities (DLTs) and determines the maximum tolderated dose (MTD).
  • NDL Non-Hodgkins lymphoma
  • DLTs dose-limiting toxicities
  • MTD maximum tolderated dose
  • COMPOUND A has low nM IC50s against SYK and JAK, decreases proliferation in ibrutinib-resistant cell lines, and suppresses tumor growth in rodent xenograft models of DLBCL.
  • Example 349 demonstrates that COMPOUND A, at dose levels associated with clinical and biomarker activity, is safe and tolerable for development as single agent and in combination with other treatments.
  • COMPOUND A The safety profile of COMPOUND A differentiates favorably from approved JAK inhibitors ruxolitinib and tofacitinib.
  • checkpoint inhibitors PD-1 / PD-1 ligand directed
  • Checkpoint inhibitors can include co-inhibitory molecules such as cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), PD-1, lymphocyte-activation gene 3, and T-cell immunoglobulin mucin-3, and co-stimulatory molecules such as:
  • glucocorticoid-induced tumor necrosis factor receptor and OX40 CD134, TNFRSF4, tumor necrosis factor receptor superfamily member 4
  • Other immune modulators that can be combined with a compound of Formula (I) include indoleamine (2,3)-dioxygenase (IDO) inhibitors, vaccines and agents that target T-cell receptors (TCR agents).
  • TCR agents can include (without limitation) chimeric antigen receptor (CAR) T cells, and TCR agonist or antagonist peptides.
  • the vaccines can include (without limitation) sipuleucel-T (Provenge®) and Talimogene laherparepvec (Imlygic®), aslo known as T-Vec.
  • COMPOUND A can be thus be clinically developed in combination with checkpoint inhibitors, IDO inhibitors, vaccines (for example, sipuleucel-T (Provenge®) and/or Talimogene laherparepvec
  • T-Vec TCR agents
  • CAR T cells and/or TCR agonist or antagonist peptides TCR agents
  • other immune-modulators to improve outcome of treatment.
  • IDO inhibitors vaccines
  • vaccines for example, sipuleucel-T (Provenge®) and/or Talimogene laherparepvec (Imlygic®), aslo known as T-Vec
  • TCR agents for example CAR T cells and/or TCR agonist or antagonist peptides
  • COMPOUND A could also improve immune mediated toxicities associated with administration of of such treatments.
  • Example 349 demonstrates the safety, tolerability, and preliminary efficacy of COMPOUND A in subjects with relapsed/refractory diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL) and mantle cell lymphoma (MCL).
  • Example 349 also demonstrates pharmacokinetic (PK) profile of COMPOUND A after single and multiple doses and the effects of COMPOUND A on Phospho-STAT3, Phospho-S6, Phospho-SYK
  • p2-microglobulin 525/526, p2-microglobulin, ⁇ , VCAM-1, TNFR2, C - reactive protein (CRP), and IL- 18.
  • Rule-based design (3+3) is used in the dose escalation component (Part A of Example 349). This method is commonly used in Phase 1 oncology clinical trials for cytotoxic agents and molecularly targeted agents. In Part A of the study, lymphomas and solid tumor patients were evaluated. The PK properties of COMPOUND A were evaluated after single and multiple dose administrations at different dose levels. Both twice and once-daily administration schedules were evaluated.
  • Expansion cohorts include patients with Diffuse Large B-Cell Lymphoma (DLBCL), Follicular cell Lymphoma FL and Mantle Cell Lymphoma (MCL) treated daily at a dose regimen to identified in Part A. Patients diagnosed with other hematologic malignancies may also be included such as T-cell lymphomas, myelofibrosis, chronic lymphocytic leukemia. Cohort expansion after determination of the MTD is commonly used in Phase 1 oncology study designs.
  • DLBCL Diffuse Large B-Cell Lymphoma
  • MCL Mantle Cell Lymphoma
  • Example 349 Additional studies for Example 349 may be conducted to characterize safety and efficacy of COMPOUND A in additional hematologic malignancies where JAK or SYK signal inhibition could provide clinical benefit. Additional studies to characterize the ADME and metabolism of COMPOUND A maybe conducted.
  • Example 349 demonstrates a Phase 1/2 clinical trial in patients with solid tumors and hematologic malignancies evaluates QD and BID oral COMPOUND A at escalating doses of 10, 20, 30, 40, 50 and 75 mg BID and 80 and 120 mg QD mg (NCT02440685).
  • Phase 1 allows patients with solid tumors or hematologic malignancies;
  • Phase 2 allows only patients with diffuse large B-Cell lymphoma (DLBCL), follicular lymphoma (FL) or mantle cell lymphoma (MCL).
  • Endpoints include safety, tolerability, pharmacokinetics, serum markers of inflammation, and response using RECIST or Lugano Classification System. Table 40 below shows clinical safety data for ongoing Example 349 (up to
  • Stable disease in a patient with primary peritoneal cancer, about 50% reduction in target lesions at 3 months in a FL patient (Lugano, 6 prior lines) and stable disease and reduction of pruritus in a peripheral T-Cell lymphoma patient after 2 months (Lugano, 2 prior lines) of treatment have been observed.
  • COMPOUND A is safe and well tolerated and does not appear to be associated with the significant thrombocytopenia, anemia and neutropenia as reported for the JAK inhibitor ruxolitinib and which should be managed by dose reduction, or interruption, or red blood cell transfusion.
  • Clinical trials report thrombocytopenia, anemia and neutropenia in 70%, 96% and 19% of patients treated with ruxolitinib.
  • COMPOUND A-related diarrhea has been reported, while the BTK signal inhibitor ibrutinib has been associated with diarrhea in over 50% of patients treated in clinical trials and severe/fatal bleeding events in up to 6%.
  • COMPOUND A is also does not appear to be associated with lipid elevation as reported for JAK inhibitor tofacitinib.
  • Clinical trials LDL elevations were reported in 15-19% of patients treated with tofacitinib.
  • Checkpoint inhibitors may serve to increase a baseline T-cell-specific immune response that turns the immune system against the tumor.
  • a disruption in the functioning of immune checkpoint molecules can lead to imbalances in immunologic tolerance that result in an unchecked immune response. This may clinically manifest with autoimmune-like/ inflammatory side-effects, which cause collateral damage to
  • immune-related adverse events are termed immune-related adverse events and are also thought to be principally T-cell mediated.
  • Other immune cells may play a role in the development of immune-related adverse events, including B cells that secrete antibodies that may mediate toxicity, granulocytes that secrete inflammatory mediators, and cytokines.
  • Patients treated with the CTLA-4 PD-1 / PD-L1 directed treatment can experience the following adverse reactions; fatigue, pruritus, diarrhea, decreased appetite, rash, pyrexia, cough, dyspnea, musculoskeletal pain, constipation, and nausea. Warnings and precautions also include immune-mediated pneumonitis, colitis, and other immune mediated adverse reactions.
  • CAR T cells have shown to be associated with similar immune-related adverse reactions and the use of CAR T cells is limited by potentially severe and fatal toxicities. CAR T cells can potentially damage normal tissues by specifically targeting a tumor-associated antigen that is also expressed on those tissues.
  • CRS cytokine release syndrome
  • CRS a systemic inflammatory response caused by cytokines released by infused CAR T cells can lead to widespread reversible organ dysfunction. CRS is the most common type of toxicity caused by CAR T cells.
  • COMPOUND A does not appear to overlap with checkpoint inhibitors, small molecule inhibitors, TCR agents (for example, CAR T cells and/or TCR agonist or antagonist peptides), vaccines (for example, sipuleucel-T (Provenge®) and/or
  • Example 349 demonstrates that COMPOUND A is safe and well tolerated. Encouraging preliminary evidence of efficacy in NHL patients has been observed. MTD was not reached.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne de nouvelles combinaisons de composés pyrimido-pyridazinone et des procédés de préparation et d'utilisation de ces combinaisons. Ces composés sont utiles pour traiter le cancer, la croissance de tumeurs ou les métastases chez des patients par administration d'une ou de plusieurs des combinaisons à un patient. Selon un mode de réalisation, la nouvelle combinaison comprend un composé de formule (I) et R1 et R2 sont définis dans la description ainsi qu'un agent immunothérapeutique.
PCT/US2018/025841 2017-04-03 2018-04-03 Combinaisons de composés de pyrimido-pyridazinone, procédés, kits et formulations associées WO2018187294A1 (fr)

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CN112043696A (zh) * 2020-08-26 2020-12-08 深圳市第二人民医院(深圳市转化医学研究院) Ido-1抑制剂在制备治疗骨性关节炎药物中的应用
US20220152065A1 (en) * 2019-08-14 2022-05-19 Rigel Pharmaceuticals, Inc. Method of blocking or ameliorating cytokine release syndrome
WO2023122662A1 (fr) * 2021-12-22 2023-06-29 The Regents Of The University Of California Inhibiteurs de liaison covalente des mutants g12s, g12d et/ou g12e de k-ras gtpase
WO2023164024A1 (fr) * 2022-02-23 2023-08-31 Teva Pharmaceuticals International Gmbh Formes à l'état solide de gusacitinib
WO2025020171A1 (fr) * 2023-07-27 2025-01-30 Libertas Bio, Inc. Formes cristallines d'un inhibiteur de jak/syk

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US20130023523A1 (en) * 2011-07-19 2013-01-24 Abbott Laboratories Pyridazino[4,5-d]pyrimidin-5(6h)-one inhibitors of kinases
US20130053346A1 (en) * 2011-08-23 2013-02-28 Endo Pharmaceuticals Inc Pyrimido-pyridazinone compounds and methods of use thereof
WO2016024228A1 (fr) * 2014-08-11 2016-02-18 Acerta Pharma B.V. Combinaisons thérapeutiques associant un inhibiteur de btk, un inhibiteur de pi3k, un inhibiteur de jak-2, un inhibiteur de pd-1 et/ou un inhibiteur de pd-l1

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US20110117073A1 (en) * 2009-09-16 2011-05-19 Avila Therapeutics, Inc. Protein Kinase Conjugates and Inhibitors
US20130023523A1 (en) * 2011-07-19 2013-01-24 Abbott Laboratories Pyridazino[4,5-d]pyrimidin-5(6h)-one inhibitors of kinases
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WO2016024228A1 (fr) * 2014-08-11 2016-02-18 Acerta Pharma B.V. Combinaisons thérapeutiques associant un inhibiteur de btk, un inhibiteur de pi3k, un inhibiteur de jak-2, un inhibiteur de pd-1 et/ou un inhibiteur de pd-l1

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20220152065A1 (en) * 2019-08-14 2022-05-19 Rigel Pharmaceuticals, Inc. Method of blocking or ameliorating cytokine release syndrome
CN112043696A (zh) * 2020-08-26 2020-12-08 深圳市第二人民医院(深圳市转化医学研究院) Ido-1抑制剂在制备治疗骨性关节炎药物中的应用
WO2023122662A1 (fr) * 2021-12-22 2023-06-29 The Regents Of The University Of California Inhibiteurs de liaison covalente des mutants g12s, g12d et/ou g12e de k-ras gtpase
WO2023164024A1 (fr) * 2022-02-23 2023-08-31 Teva Pharmaceuticals International Gmbh Formes à l'état solide de gusacitinib
WO2025020171A1 (fr) * 2023-07-27 2025-01-30 Libertas Bio, Inc. Formes cristallines d'un inhibiteur de jak/syk

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