NZ720909B2 - 2-amino-6-fluoro-n-[5-fluoro-pyridin-3-yl]pyrazolo[1,5-a]pyrimidin-3-carboxamide compound useful as atr kinase inhibitor, its preparation, different solid forms and radiolabelled derivatives thereof - Google Patents

Abstract

The present invention relates to compounds useful as inhibitors of ATR protein kinase. The invention relates to pharmaceutically acceptable compositions comprising the compounds of this invention; methods of treating of various diseases, disorders, and conditions using the compounds of this invention; processes for preparing the compounds of this invention; intermediates for the preparation of the compounds of this invention; and solid forms of the compounds of this invention. The compounds of this invention have formula I-A or I-B: wherein the variables are as defined herein. n; processes for preparing the compounds of this invention; intermediates for the preparation of the compounds of this invention; and solid forms of the compounds of this invention. The compounds of this invention have formula I-A or I-B: wherein the variables are as defined herein.

Description

-AMINOFLUORO-N-[5-FLUORO-PYRIDINYL]PYRAZOLO[1,5-A]PYRIMIDINCARBOXAMIDE COMPOUND USEFUL AS ATR KINASE INHIBITOR, ITS PREPARATION, DIFFERENT SOLID FORMS AND RADIOLABELLED DERIVATIVES THEREOF BACKGROUND OF THE INVENTION ATR (“ATM and Rad3 related”) kinase is a protein kinase involved in cellular responses to DNA damage. ATR kinase acts with ATM (“ataxia telangiectasia mutated”) kinase and many other proteins to regulate a cell’s response to DNA damage, commonly referred to as the DNA Damage se (“DDR”). The DDR stimulates DNA repair, promotes al and stalls cell cycle progression by activating cell cycle checkpoints, which e time for repair. Without the DDR, cells are much more sensitive to DNA damage and readily die from DNA lesions induced by endogenous cellular processes such as DNA replication or exogenous DNA damaging agents commonly used in cancer therapy. y cells can rely on a host of different proteins for DNA repair ing the DDR kinase ATR. In some cases these proteins can compensate for one another by activating functionally redundant DNA repair processes. On the contrary, many cancer cells harbour s in some of their DNA repair processes, such as ATM signaling, and therefore display a greater reliance on their remaining intact DNA repair proteins which include ATR.

In addition, many cancer cells express activated oncogenes or lack key tumour suppressors, and this can make these cancer cells prone to dysregulated phases of DNA ation which in turn cause DNA damage. ATR has been implicated as a critical component of the DDR in response to disrupted DNA replication. As a result, these cancer cells are more ent on ATR activity for survival than healthy cells. Accordingly, ATR inhibitors may be useful for cancer treatment, either used alone or in combination with DNA damaging agents, because they shut down a DNA repair ism that is more important for cellular al in many cancer cells than in healthy normal cells.

In fact, disruption of ATR function (e.g. by gene deletion) has been shown to promote cancer cell death both in the absence and presence of DNA damaging agents. This suggests that ATR inhibitors may be effective both as single agents and as potent sensitizers to radiotherapy or genotoxic chemotherapy.

For all of these reasons, there is a need for the development of potent and selective ATR tors for the ent of cancer, either as single agents or as combination therapies with radiotherapy or genotoxic chemotherapy. rmore, it would be desirable to have a synthetic route to ATR inhibitors that is amenable to large-scale synthesis and improves upon currently known methods.

ATR peptide can be expressed and isolated using a y of methods known in the literature (@ e.g., Unsal—Kacmaz et al, PNAS 99: 10, pp6673—6678, May 14, 2002; see also Kumagai et al. Cill 124, pp943-955, March 10, 2006; Unsal-Kacmaz et al. Molecular and Cellular m,Feb 2004, p1292-1300; and Hall—Jackson et al. Oncogene 1999, 18, 6707—6713).

BRIEF DESCRIPTION OF THE FIGURES FIGURE 1a: XRPD nd I-I ethanol solvate FIGURE 2a: TGA nd I-1° ethanol solvate FIGURE 3a: DSC Compound I ethanol solvate FIGURE 4a: solid state 13C NMR spectrum (12.5 kHz ng) of Compound I-1° ethanol solvate FIGURE 5a: solid state 19F NMR spectrum (12.5 kHz spinning) of Compound I-1° l solvate FIGURE 1b: XRPD Compound I-1° e I FIGURE 2b: TGA Compound I-1° hydrate I FIGURE 3b: DSC Compound I hydrate I FIGURE 4b: XRPD Compound I-1° hydrate II FIGURE 5b: solid state 13C NMR spectrum (11 kHz spinning) of Compound I-1° hydrate II FIGURE 6b: solid state 19F NMR spectrum (11 kHz spinning) of Compound I-1° hydrate II FIGURE 1c: XRPD Compound I-I anhydrous form A FIGURE 2c: TGA Compound I-I anhydrous form A FIGURE 3c: DSC Compound I-I anhydrous form A FIGURE 4c: is a conformational plot of Compound I-1° anhydrous form A based on single crystal X—ray analysis.

FIGURE 5c: is a conformational plot showing the stacking order of Compound I-1° ous form FIGURE 6c: solid state 13C NMR spectrum (12.5 kHz spinning) of Compound I-1° anhydrous form FIGURE 7c: solid state 19F NMR spectrum (12.5 kHz spinning) of Compound I-1° anhydrous form FIGURE 1d: XRPD Compound I-1° anhydrous form B FIGURE 2d: TGA Compound I-1° anhydrous form B FIGURE 3d: DSC Compound I anhydrous form B FIGURE 4d: solid state 13 C NMR spectrum (12.5 kHz spinning) of nd I-1° anhydrous form FIGURE 5d: solid state 19F NMR spectrum (12.5 kHz spinning) of Compound I-1° anhydrous form FIGURE 1e: XRPD Compound I-1° anhydrous form C FIGURE 2e: TGA Compound I-1° anhydrous form C FIGURE 3e: DSC Compound I-1° anhydrous form C FIGURE 4e: solid state 13C NMR spectrum (12.5 kHz spinning) of Compound I-1° anhydrous form FIGURE 5e: solid state 19F NMR spectrum (12.5 kHz spinning) of Compound I-1° anhydrous form FIGURE 1f: XRPD Compound I-1° amorphous form FIGURE 2f: DSC Compound I-1° ous form FIGURE 3f: solid state 13C NMR spectrum (12.5 kHz spinning) of Compound I-1° ous FIGURE 4f: solid state 19F NMR um (12.5 kHz spinning) of nd I-1° amorphous FIGURE lg: XRPD Compound I-1° DMSO e FIGURE 2g: TGA Compound I-1° DMSO solvate FIGURE 3g: DSC Compound I-1° DMSO solvate FIGURE lh: XRPD Compound I-1° DMAC solvate FIGURE 2h: TGA Compound I-1° DMAC solvate FIGURE 3h: DSC Compound I-1° DMAC solvate FIGURE 1i: XRPD Compound I-1° acetone solvate FIGURE 2i: TGA Compound I-1° acetone solvate FIGURE 3i: DSC Compound I-1° acetone solvate FIGURE lj: XRPD Compound I-1° isopropanol e FIGURE 2j: TGA Compound I-1° isopropanol solvate FIGURE 3j: DSC Compound I-1° isopropanol solvate SUMMARY OF THE INVENTION The present invention relates to solid forms ofATR inhibitors, compositions including ATR inhibitors, as well as deuterated ATR inhibitors. The present invention also relates to processes and intermediates for preparing nds useful as inhibitors of ATR kinase, such as amino- pyrazolopyrimidine derivatives and related molecules. Amino-pyrazolopyrimidine derivatives are useful as ATR inhibitors and are also useful for ing ATR inhibitors.

One aspect of the invention es a process for preparing a compound of formula I-A: Another aspect comprises a process for preparing a compound of formula 1-1: 0 {FM r aspect of the present invention comprises a compound of formula I-B: or a pharmaceutically able salt or derivative thereof, wherein: each Y1 Y2 Y3 Y4 Y5 Y6 Y7 Y8 Y9 Y10 Y“ Y12 YB Y14 Y15 Yrs Y17 Y18 and Y19 is independently hydrogen or deuterium; provided at least one of Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10,Y11,Y12, Y”, Y14,Y15,Y16,Y17,Y18, and Y” is deuterium each X1, X2, X4, X5, X6, X7, X8, and X9 is independently selected from 12C or 13C; and X3 is independently selected from -12C(O)- or -13C(O)-.

Yet another aspect of the invention provides solid forms of a compound of formula I-1: I-1. [0011A] In a particular aspect, the present invention es a compound of a I-1 wherein the form is selected from the group consisting of Compound I-1 hydrate I, Compound I-1 ous form A, Compound I-1 anhydrous form B, nd I-1 anhydrous form C, Compound I-1 DMSO e, Compound I-1 DMAC solvate, Compound I-1 acetone solvate, or Compound I-1 isopropanol solvate, wherein Compound I-1 hydrate I is crystalline Compound I-1 hydrate I characterized by one or more peaks expressed in 2-theta ± 0.2 at 6.5, 12.5, 13.7, 18.8, and 26.0 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha ion, Compound I-1 anhydrous form A is lline Compound I-1 anhydrous form A characterized by one or more peaks expressed in 2-theta ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 s in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation, Compound I-1 anhydrous form B is crystalline Compound I-1 anhydrous form B characterized by two or more peaks expressed in 2-theta ± 0.2 at 7.2, 8.3, 12.9, 19.5, and 26.6 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation, Compound I-1 ous form C is crystalline Compound I-1 anhydrous form C characterized by two or more peaks expressed in 2-theta ± 0.2 at 6.8, 13.4, 15.9, 30.9, and 32.9 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation.

Some embodiments sed herein generally relate to a composition that can include an effective amount of Compound I-1 or polymorphic anhydrous form A of Compound I-1. (hereinafter “Form A”), or a pharmaceutically acceptable salt of the aforementioned compounds. [0012A] In a particular aspect, the t invention provides a composition comprising: (followed by page 5a) a) nd I-1, or a pharmaceutically acceptable salt thereof, wherein nd I-1 is represented by the following structural formula: ; and b) one or more excipients, wherein at least 90% by weight of Compound I-1 is crystalline Compound I-1•anhydrous form A characterized by one or more peaks expressed in 2-theta ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X-Ray powder diffraction n obtained using Cu K alpha radiation. [0012B] In another particular aspect, the present ion provides a crystal form of Compound I-1 having a monoclinic crystal system, a P21/c centrosymmetric space group, and the following unit cell parameters: a = 15.29(3)Å α = 90° b = 12.17(2)Å β = 107.22(3)° c = 14.48(3)Å γ = 90°, wherein nd I-1 is represented by the following structural formula: ] In yet another particular aspect, the present invention provides a process for preparing Compound I-1•anhydrous form A comprising ng a suspension containing Compound I- 1•ethanol solvate and tetrahydrofuran, wherein Compound I-1•anhydrous form A is crystalline Compound I-1•anhydrous form A characterized by one or more peaks expressed in 2-theta ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha ion, wherein Compound I-1 is represented by the following structural formula: 5a (followed by page 5b) [0012D] In a further particular aspect, the present invention provides a process for preparing Compound I-1•anhydrous form A comprising stirring a suspension containing Compound I- 1•amorphous, isopropanol, and water, wherein Compound I-1•anhydrous form A is crystalline Compound I-1•anhydrous form A terized by one or more peaks expressed in 2-theta ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation, wherein nd I-1 is represented by the following structural formula: Other embodiments sed herein generally relate to a method of preparing such compositions bed herein (for example, a composition that can include an effective amount of Compound I-1 or Form A, or a pharmaceutically acceptable salt of the entioned compounds).

Still other embodiments disclosed herein generally relate to a method of treating cancer using a composition described herein.

Some embodiments disclosed herein generally relate to the use of a composition bed herein (for example, a composition that includes an effective amount of Compound I-1 or Form A, or a pharmaceutically acceptable salt of the aforementioned compounds) in the cture of a medicament for treating cancer.

Other aspects of the invention are set forth herein. 5b (followed by page 5c) DETAILED DESCRIPTION OF THE INVENTION For purposes of this application, it will be understood that the terms embodiment,example, and aspect are used interchangeably.

Processes Another aspect of the present invention comprises a process for ing a compound of formula I-A: [FOLLOWED BY PAGE 6] 2014/068713 under suitable conditions to form an amide bond, wherein: R1 is independently selected from fluoro, chloro, or —C(J1)2CN; J1 is independently selected from H or C1_2alkyl; or two occurrences of J1 her with the carbon atom to which they are attached, form a 3—4 membered ally substituted carbocyclic ring; R2 is independently selected from H; halo; -CN; NHz; a C1_2alkyl optionally substituted with 0—3 occurrences of fluoro; or a C1_3aliphatic chain wherein up to two methylene units of the aliphatic chain are ally replaced with —O—, —NR—, —C(O)—, or —S(O)n; R3 is independently selected from H; halo; C1_4alkyl optionally substituted with 1-3 occurrences of halo; C3_4cycloalkyl; -CN; or a C1_3aliphatic chain n up to two methylene units of the aliphatic chain are optionally replaced with —O-, -NR—, —C(O)—, or — S(O)n; R4 is independently selected from Q1 or a C1_10aliphatic chain wherein up to four methylene units of the aliphatic chain are optionally replaced with —O—, —NR—, —C(O)—, or —S(O)n—; each R4 is optionally substituted with 0—5 occurrences of JQ; or R3 and R4, taken together with the atoms to which they are bound, form a 5—6 membered aromatic or non-aromatic ring haVing 0-2 heteroatoms ed from oxygen, nitrogen or sulfur; the ring formed by R3 and R4 is optionally substituted with 0—3 ences of JZ; Q1 is independently ed from a 3—7 membered fully saturated, partially unsaturated, or aromatic monocyclic ring, the 3-7 membered ring having 0—3 heteroatoms selected from oxygen, nitrogen or ; or an 7-12 membered fully saturated, partially rated, or aromatic bicyclic ring having 0-5 atoms selected from oxygen, nitrogen, or sulfur; JZ is independently selected from C1_6aliphatic, :0, halo, or ->O; JQ is independently selected from —CN; halo; =0; Q2; or a C1_galiphatic chain wherein up to three methylene units of the aliphatic chain are optionally replaced with —O-, -NR-, -C(O)- or —S(O)n—; each occurrence of JQ is ally substituted by 0—3 occurrences of JR; or two occurrences of J on the same atom, taken together with the atom to which they are joined, form a 3-6 membered ring haVing 0-2 heteroatoms ed from oxygen, nitrogen, Q is optionally substituted with or sulfur; wherein the ring formed by two occurrences of J 0—3 occurrences of JX; or two occurrences of JQ, together with Q, form a 6—10 membered ted or partially unsaturated bridged ring system; Q2 is independently selected from a 3—7 ed fully saturated, partially unsaturated, or aromatic monocyclic ring haVing 0-3 heteroatoms selected from , nitrogen, or sulfur; or an 7-12 membered fully saturated, partially unsaturated, or aromatic bicyclic ring haVing 0-5 heteroatoms ed from oxygen, nitrogen, or sulfur; JR is independently selected from —CN; halo; =0; ->O; Q3; or a C1_6aliphatic chain wherein up to three methylene units of the aliphatic chain are optionally replaced with —O-, -NR-, - C(O)—, or —S(O)n—; each JR is optionally substituted with 0—3 occurrences of JT; or two occurrences of J on the same atom, together with the atom to which they are joined, form a 3-6 membered ring haVing 0-2 heteroatoms selected from oxygen, nitrogen, or sulfur; wherein the ring formed by two occurrences of JR is optionally substituted with 0—3 occurrences of JX; or two occurrences of JR, together with Q2, form a 6—10 membered saturated or partially unsaturated bridged ring system; Q3 is a 3-7 membered fully saturated, partially unsaturated, or aromatic monocyclic ring haVing 0-3 heteroatoms selected from oxygen, nitrogen, or sulfur; or an 7-12 membered fully saturated, partially unsaturated, or aromatic bicyclic ring haVing 0-5 heteroatoms selected from oxygen, nitrogen, or sulfur; JX is independently selected from-CN; :0; halo; or a C1_4aliphatic chain wherein up to two methylene units of the tic chain are optionally replaced with —O-, -NR—, , or — S(O)n—; JT is ndently ed from halo, -CN; ->O; =0; —OH; a C1_6aliphatic chain wherein up to two methylene units of the tic chain are ally replaced with —O-, -NR-, - C(O)—, or —S(O)n—; or a 3—6 membered non—aromatic ring having 0—2 heteroatoms selected from oxygen, en, or sulfur; each occurrence of IT is optionally substituted with 0—3 occurrences of JM; or two occurrences of IT on the same atom, together with the atom to which they are joined, form a 3-6 membered ring having 0-2 heteroatoms selected from oxygen, nitrogen, or sulfur; or two occurrences of IT, together with Q3, form a 6—10 membered saturated or partially unsaturated bridged ring system; JM is independently selected from halo or C1_6aliphatic; J is H or Cl; n is 0, l or 2; and R is independently selected from H or C1_4aliphatic.

For purposes of this application, it will be tood that when two occurrences of IQ, together with Q, form a bridged ring system, the two occurrences of IQ are attached to separate atoms of Q1. Additionally, when two occurrences of JR, er with Q2, form a bridged ring system, the two occurrences of JR are attached to separate atoms of Q2. Moreover, when two ences of IT, together with Q3, form a bridged ring system, the two occurrence of IT are attached to separate atoms of Q3.

It will be understood by those skilled in the art that the arrow in ->O ents a dative bond. on Conditions In some examples, the suitable conditions for forming the amide bond comprises reacting the compound of formula 6 with a substituted 3-amino pyridine in an aprotic solvent under heat. In other es, the aprotic solvent is ed from NMP, optionally substituted pyridine, or DMF.

In another embodiment, the aprotic solvent is optionally substituted pyridine. In still other embodiments, the on temperature is at least 80°C. In another embodiment, the reaction temperature is at least 100°C.

In r embodiment, the process, described above, further comprises ing a compound of formula 6: by reacting a compound of formula 5: \MOH under suitable conditions to form an activated ester, wherein R1 and J are as defined .

In some embodiments, suitable conditions for forming the activated ester comprises reacting the compound of formula 5 with an amide coupling agent in the ce of an organic base.

In other embodiments, the organic base is an tic amine. In still other embodiments, the organic base is independently selected from triethylamine or DIPEA. In one or more embodiments, the amide coupling agent is independently selected from TBTU, TCTU, HATU, T3P, or COMU. In yet another embodiment, the amide coupling agent is ndently selected from TBTU or TCTU. In another embodiment, the amide ng agent is TCTU.

Another aspect of the invention comprises a process for preparing a compound of formula I-A: NH2 0 N / R2 "i / / N \ l N H R3 g\ //N R4 comprising reacting a compound of formula 5: NH2 0 \MOH under suitable conditions to form an amide bond, wherein R1, R2, R3, and R4 are as defined herein.

Yet another apect of the t invention comprises a process for ing a nd of formula 5: by reacting a compound of formula 4: NH2 0 "l / / O,AII under suitable hydrolytic conditions, wherein R1 is as defined herein.

In some embodiments, suitable hydrolytic conditions comprise reacting the compound of formula 4 with a silane in the presence of a metal st. In other embodiments, the silane is a phenylsilane. In another ment, the metal catalyst is a palladium catalyst. In yet another embodiment, the palladium catalyst is Pd(PPh3)4. In another embodiment suitable hydrolytic conditions comprise reacting the compound of formula 4 with ylbenzenesulf1nate in the presence of a metal catalyst.

In still other embodiments, suitable hydrolytic conditions comprise reacting the compound of formula 4 with an aqueous alkali. In some embodiments, the aqueous alkali is selected from LiOH, NaOH or KOH.

Another aspect of the present invention comprises a process for preparing a compound of formula 4: by reacting a compound of formula 3: NH2 0 N\/ / O,AII under suitable condensation conditions to form a pyrimidine ring.

In some embodiments, suitable sation conditions to form a pyrimidine ring se reacting the compound of formula 3 with a 1,3-dielectrophilic species in the presence of a solvent. In another embodiment, the 1,3—dielectrophilic species is ed from 1,3—dialdehyde or 3-(dialkylamino)-propenal. In still other ments, the solvent is selected from DMF or DMSO. In other ments, the 1,3—dielectrophilic species is generated in situ from a protected 1,3-dielectrophilic species. In another embodiment, the 1,3-dielectrophilic species is generated from a ketal in the presence of a sulfonic acid. In yet another embodiment, the ic acid is PTSA.

Another aspect of the present invention comprises a process for preparing the compound offormula3: NH2 0 "l / / O,AII by reacting a compound of formula 2: H2N col3 under suitable condensation conditions to form a pyrazole ring.

In some embodiments, suitable condensation conditions to form a pyrazole ring comprise reacting the compound of formula 2 with a hydrazine or hydrazine hydrate in the presence of an aprotic solvent under basic conditions. In another embodiment, the aprotic solvent is DMF. In yet another ment, the basic conditions comprise reacting the compound of formula 2 in the ce of potassium acetate or sodium acetate.

Yet another aspect of the present invention comprises a process for ing a compound of formula 2: NCfiO’AII H2N CCI3 by reacting a compound of formula 1: NC\)J\OAII under suitable anion condensation conditions.

In some embodiments, suitable anion condensation conditions comprise l) reacting the compound of formula 1 with a base, in the ce of a solvent, to generate the anion of the compound of formula 1; and 2) reacting the anion of the compound of formula 1 with trichloroacetonitrile. In still other embodiments, the base is potassium acetate. In yet another embodiment, the solvent is an alcohol. In other embodiments, the solvent is isopropylalcohol.

One embodiment of the t invention comprises a process for ing a compound of formula I-A: NH2 0 /N R2 "l / / N \ / N H R3 g\ R4 //N sing reacting a compound of formula 9: NH2 0 /N R2 N\ / / N \ / HN H R3 NH2 R4 under suitable condensation conditions to form a pyrimidine ring, wherein R1, R2, R3 and R4 are as defined .

In some embodiments, suitable condensation conditions to form a pyrimidine ring comprise reacting the compound of formula 9 with a 1,3-dielectrophilic species in the presence of a solvent. In another embodiment, the 1,3—dielectrophilic species is selected from 1,3—dialdehyde or 3-(dialkylamino)-prop-2—enal. In still other embodiments, the solvent is selected from DMF or DMSO in water. In other embodiments, the electrophilic species is generated in situ from a protected 1,3—dielectrophilic species. In another embodiment, the 1,3—dielectrophilic species is generated from a ketal in the presence of a sulfonic acid. In yet another embodiment, the sulfonic acid is PTSA. r embodiment of the present invention comprises a process for ing a compound of formula 9: by reacting a compound of formula 8: NMH R3 under suitable condensation conditions to form a le ring.

In some ments, suitable sation conditions to form a pyrazole ring comprise l) reacting the compound of formula 8 with a base, in the presence of a t, to generate the anion of the compound of formula 8; 2) reacting the anion with trichloroacetonitrile; and 3) reacting the product from 2) with a hydrazine or hydrazine hydrate in the ce of an aprotic solvent. In another embodiment, the aprotic solvent is NMP or DMF. In some embodiments, the base is selected from sodium acetate or potassium acetate.

Yet another ment comprises a process for preparing a compound of formula 8: O N 2 / R NC\AH/g/l/ by reacting a compound of formula 7: NC\/[( under le conditions to form an amide bond.

In some examples, the suitable conditions for forming the amide bond ses reacting the compound of formula 7 with a substituted 3-amino ne with an amide ng agent in the presence of an c solvent and an organic base. In other examples, the aprotic solvent is selected from NMP or DMF. In another embodiment, the organic base is an aliphatic amine. In still other embodiments, the organic base is independently selected from triethylamine or DIPEA. In yet another embodiment, the amide coupling agent is independently selected from TBTU or TCTU.

Synthesis of Compound 1-1 Another aspect of the present invention provides a process of preparing a compound of formula 1-1: comprising the step of reacting the compound of formula 30 NH2 0 /N N N <\ /N .HCl O OH with a compound of formula 25: 2014/068713 under suitable conditions to form an amide bond.

Still other embodiments of the present invention comprise provides a process for preparing the compound of formula 30: %QNH2 0 / 209IN by ng the compound of formula 28: under suitable deprotection conditions to form the carboxylic acid.

Another embodiment provides a process for preparing a compound of formula 28: NHZO /| N\// N\ F N N [>\//N o o by reacting the compound of formula 621*: NH2 0 N:N N / 0’ MN CI with a compound of formula 27: H2N F under suitable conditions to form an amide bond.

In some embodiments, le conditions for forming the amide bond comprise reacting the compound of formula 30 with the compound of formula 25 in the presence of an amide coupling partner, an aprotic solvent, and a base. In other embodiments, the aprotic solvent is independently selected from NMP, DMF, or tetrahydrofuran. In still other embodiments, the aprotic t is tetrahydrofuran. In another embodiment, the base is an aliphatic amine. In yet another embodiment, the base is DIPEA. In some embodiments, the amide coupling partner is independently ed from CDI, TBTU or TCTU. In one or more embodiments, the amide ng partner is TCTU. In yet another embodiment, the amide coupling partner is CD1.

In other embodiments, suitable deprotection ions comprise reacting the compound of formula 28 with an acid in the presence of a solvent. In some embodiments, the acid is HCl. In another embodiment, the solvent is l,4-dioxane.

In yet another embodiment, le conditions for forming the amide bond comprise reacting the compound of formula 621* with the nd of formula 27 in an aprotic solvent under heat. In still other embodiments, the aprotic solvent is independently ed from NMP, pyridine, or DMF. In another ment, the aprotic solvent is pyridine. In some embodiments, the reaction is carried out at a temperature of at least 80°C.

Another aspect of the present invention provides a process of preparing a compound of formula 27: F/NH2 comprising the step of reacting a compound of formula 26: F /Br 09.x under le conditions to form an amine.

In some embodiments, suitable conditions to form an amine comprise reacting the compound of formula 27 under Buchwald-Hartwig amination conditions, known to those skilled in the art.

Yet another embodiment provides a process for ing a compound of formula 26: F Br 0 0k by 1) reacting a nd of formula 18: F Br under suitable halogen exchange ions to generate the compound of formula 32 F Br 32 and 2) reacting the compound of formula 32: F Br with a compound of formula 22: 0 OX under suitable displacement conditions.

In some embodiments, suitable halogen exchange conditions comprise reacting the compound of formula 18 with potassium fluoride in the presence of an aprotic t and a phase er catalyst. In other embodiments, the aprotic solvent is independently selected from DMSO, DMF, or sulfolane. In still other embodiments, the phase transfer catalyst is Me4NCl. In still other embodiments, le displacement conditions comprise reacting the compound of formula 32 with a nd of a 22 in the presence of a base. In another embodiment, the base is an aliphatic amine. In some embodiments, the aliphatic amine is DIPEA.

Other embodiments of the present invention provides a process for preparing a nd of formula 18: by reacting the compound of formula 31: under suitable halogenation conditions.

In some embodiments, suitable halogenation conditions comprise l) reacting the compound of formula 31 with a base to generate an anion; and 2) reacting the anion with a chlorinating agent. In yet r embodiment, the base is LDA. In another embodiment, the chlorinating agent is l, l , l,2,2,2-hexachloroethane.

Some embodiments of the present invention provides a process for ing a compound of formula I-1: 3%;NH2 0 / ONTO comprising the step of reacting the compound of formula 33: with a compound of formula 25: under suitable conditions to form an amide bond.

In some embodiments, suitable conditions for forming the amide bond comprise reacting the compound of formula 33 with the compound of formula 25 in the presence of an amide ng partner, an aprotic solvent, and a base. In other ments, the aprotic solvent is ndently selected from NMP, DMF, or tetrahydrofuran. In still other embodiments, the aprotic solvent is tetrahydrofuran. In another embodiment, the base is an tic amine. In yet another embodiment, the base is DIPEA. In some embodiments, the amide coupling partner is independently selected from TBTU or TCTU. In one or more embodiments, the amide coupling partner is TCTU.

Yet another embodiment es a process for preparing a compound of formula 33: comprising the step of reacting the compound of formula 28: NHZO /| M V F N N [>\ //N O O under suitable deprotecting conditions.

In some embodiments, le ecting conditions for ng the tert-butyl ester comprise reacting the compound of formula 28 with an acid in the presence of a solvent. In one embodment, the acid is selected from, but not limited to, methanesulphonic acid (preferred), PTSA, TFA, or HCl. In still other embodiments, the solvent is selected from, but is not limited to, 1,4- dioxane or acetonitrile. In another embodiment, the t is acetonitrile.

Another embodiment provides a process for preparing a compound of formula 43: comprising the steps of: a) reacting a compound of formula 35: RMOWCLR" ,O O\ R° R° wherein R0 is C1_6aliphatic, under acidic conditions to form a compound of formula 36: HOVH b) reacting a compound of formula 36 with an electrophilic fluorinating agent to form a compound of a 38: HOVH c) reacting a compound of formula 38 with a compound of formula 3: o—\: under suitable condensation conditions to form the compound of formula 43.

In some embodiments, R0 is independently selected from methyl, ethyl, propyl, isopropyl, butyl, and pentyl. In still other embodiments, RO is independently selected from methyl or ethyl.

In r embodiment, the electrophilic fluorinating agent is l-(Chloromethyl)fluoro- l,4-diazoniabicyclo[2.2.2]octane ditetrafluoroborate. In other embodiments, the electrophilic fluorinating agent is fluorine gas.

In yet another embodiment, the suitable sation conditions comprise reacting the compound of a 38 With the compound of formula 3 in the presence of a solvent and heat. In some embodiments, the solvent is selected from DMF or DMSO.

Yet another embodiment provides a process for preparing a compound of formula I-1: gig? ONTO sing the steps of: a) ng the compound of formula 63*: NHZO NZN W/ O <> /,N CI with a compound of formula 27: O 0% under suitable amide bond formation conditions to form a compound of formula 28: NHZO /| NH V F N N S\ //N b) purifying the compound of formula 28 using a suitable palladium tering agent; c) reacting the compound of formula 28 under suitable ection conditions to form a compound of formula 30 NHZQ /NI ”3/ n F N N <\ /N .HCl O OH ; and d) reacting the compound of formula 30 with a compound of formula 25: under suitable amide bond formation conditions to form the compound of formula 1-1.

In some embodiments, the suitable palladium sequestering agent is independently selected from propane-1,2-diamine; ethane-1,2-diamine; ethane-1,2-diamine; propane-1,3-diamine; tetramethylethelenediamine; ethylene ; l,3-bis(diphenylphosphanyl)propane; 1,4- bis(diphenylphosphanyl)butane; and s(diphenylphosphanyl)ethane/Pr-1,2-diamine. In still other embodiments, the suitable palladium sequestering agent is propane-1,2-diamine.

Another embodiment provides a process for preparing a nd of formula 28: NHZQ /N| N’/ V F N N [>\ //N o o comprising the steps of: a) ng the compound of formula Sa NH2 0 N OH under suitable halogenation conditions to form a compound of formula 34: NH2 0 N/ X wherein X is halogen; b) reacting the compound of formula 34 with a nd of formula 27: under suitable amide bond ion conditions to form a nd of formula 28.

In some embodiments X is independently selected from fluoro or chloro. In another embodiment, X is chloro. In some ments, the suitable halogenation conditions comprise reacting the compound of formula 521 with a halogenating agent and a base in the presence of a solvent. In yet another embodiment the halogenating agent is SOClz. In some embodiments, the base is triethylamine. In still other embodiments, the solvent is DCM.

Yet another aspect of the present invention provides a process for preparing a nd of formula I-l: gig? ONTO comprising the steps of: a) reacting the compound of formula Sa NH2 0 N OH 2014/068713 under suitable halogenation conditions to form a compound of formula 34: NH2 0 N/ x wherein X is halogen; b) reacting the compound of formula 34 with a compound of formula 27: O OJ< under suitable amide bond formation conditions to form a compound of formula 28: NHZO /| NH V F N N S\ //N c) reacting the compound of formula 28 under le deprotection conditions to form a compound of formula 30 NHZQ /N| N/ N F \ / H N N <\ /N .HCl 0 OH d) reacting the compound of formula 30 with a compound of formula 25: under suitable amide bond formation conditions to form the nd of formula I-1.

Deuterated Compounds In r embodiment, Isotopes can be introduced into compound 1-1 by selecting building blocks that contain the isotopic atoms (either commercially available or that can be prepared according to processes known to those skilled in the art) and engaging them into a sequence similar to the one reported for the unlabelled material.

Another aspect of the present invention provides a nd of Formula I-B: or a pharmaceutically acceptable salt thereof, wherein: each Y1 Y2 Y3 Y4 Y5 Y6 Y7 Y8 Y9 Y10 Y“ Y12 YB Y14 Y15 Yrs Y17 Y18 and Y19 is independently en or deuterium; provided at least one of Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10,Y11,Y12, Y”, Y14,Y15,Y16,Y17,Y18, and Y” is ium each X1, X2, X4, X5, X6, X7, X8, and X9 is independently selected from 12C or 13C; and X3 is independently selected from —12C(O)— or —13C(O)—.

The following labeled ng blocks, which can be used in the tic route for preparing the compound of formula I-B, are all commercially ble: 0 2,2,3,3,5,5,6,6—octadeuteropiperazine; 0 2,3,5,6-tetra-13C-piperazine; 0 2,2,3,3,4,5,5,6,6—nonadeuteropiperidine—4—carboxylic acid; 0 l, 2—Di13Ccyanoacetic acid; o 1-13Ccyano(13C)acetic acid ethyl ester; and o 2—13C—2—cyano(13C)acetic acid ethyl ester.

Other labeled building blocks, which may be utilized in the synthetic route for preparing a compound of formula I-B, are known to those skilled in the art. These may include, but are not limited to, the following labeled building blocks: 0 2—13C-oxetan—3—one; 0 3—13C—oxetan—3—one; 0 2,2,3 ,3 -tetradeuteropiperazine; 0 2,2,5,5-tetradeuteropiperazine; 0 4—deuteropiperidinecarboxylic acid ethyl ester; 0 2—cyano(13C)acetic acid; 0 1—13C—2—cyanoacetic acid; 0 2-13Ccyanoacetic acid; and 0 l-deutero-3 -(diethylamino)-2—fluoroacrylaldehyde; In one or more ments, Y”, Y13 and Y19 are deuterium, and , Y”, Y”, Y16, Y”, Y”, Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10, and Y11 are independently selected from hydrogen or deuterium. In another embodiment, Y”, Y”, Y”, Y”, Y16, Y”, Y”, and Y19 are deuterium, and Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y“), and Y“ are en.

In yet another embodiment, X1, X2, X4, X5, X6, X7, X8, and Xgare 12C; and X3 is -12C(O)-. In still other ments, X1, X4, X5, X6, X7, X8, and X9 are 12C; X3 is -13C(O)-; and X2 is 13 C.

In some embodiments, Y“, Y”, Y”, Y”, Y”, Y16, Y”, Y”, and Y19 are deuterium, and Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, and Y10 are independently selected from hydrogen or deuterium.

In other embodiments, Y“, Y”, Y”, Y”, Y”, Y16, Y17, Y18, and Y19 are deuterium, and Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, and Y10 are hydrogen.

In yet another embodiment, Y2, Y”, Y”, Y”, Y”, Y16, Y17, Y18, and Y19 are ium, and Y1, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10, and Y11 are ndently selected from hydrogen or deuterium. In another aspect of the ion, Y2, Y”, Y13 and Y19 are , Y”, Y”, Y16, Y”, Y”, deuterium, and Y1, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10, and Y11 are hydrogen.

In some embodiments, Y”, Y13 and Y19 are deuterium, and Y1, Y2, Y3, Y4, Y5, Y6, , Y18, Y7, Y8, Y9, Ylo, Y“, Y”, Y”, Y16, and Y17 are hydrogen or deuterium. In still other embodiments, Y”, Y”, Y”, and Y” are deuterium, and Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y”, Y“, Y”, Y”, Y”, and Y17 are hydrogen.

In one or more embodiments, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10, and Y11 are deuterium, and Y1, Y2, Y”, Y13 , Y”, Y”, Y”, Y”, Y”, and Y19 are independently selected from deuterium or hydrogen. In another embodiment, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10, and Y11 are deuterium, and Y1, Y2, Y”, Y”, Y”, Y”, Y”, Y”, Y”, and Y” are hydrogen.

In yet another embodiment, Y2 and Y11 are deuterium, and Y1, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Ylo, Y”, Y”, Y”, Y”, Y16, Y”, Y18, and Y19 are deuterium or hydrogen. In other embodiments, Y2 and Y11 are deuterium, and Y1, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y”, Y”, Y”, Y”, Y”, Y”, Y”, Y”, and Y19 are hydrogen.

In some embodiments, Y2 is deuterium, and Y1, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y“), Y“, Y”, Y13 and Y19 are deuterium or hydrogen. In another embodiment, Y2 is , Y”, Y”, Y16, Y”, Y”, deuterium, and Y1, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y”, Y“, Y”, Y”, Y”, Y”, Y”, Y”, Y”, and Y” are In still other embodiments, X1, X2, X4, X5, X6, X7, and X8 are 12C; X3 is -12C(O)-; and X9 is 13C. In another embodiment, X1, X2, X8, and Xgare 12C; X3 is -12C(O)-; and X4, X5, X6, and X7 are 13C. In yet another embodiment, X2, X4, X5, X6, X7, X8, and X9 are 12C; X3 is -12C(O)-; and X1 is 13C. In other embodiments, X2, X4, X5, X6, X7, and X9 are 12C; X3 is -13C(O)-; and X1 and X8 are 13C.

In some embodiments, Y11 is deuterium, and Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y“), Y”, Y13 and Y19 are ndently selected from hydrogen or deuterium. In , Y”, Y”, Y”, Y”, Y”, r embodiment, Y11 is deuterium, and Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Ylo, Y”, Y13 , Y”, Y”, Y16, Y”, Y”, and Y19 are hydrogen.

In yet another ment, X1, X4, X5, X6, X7, X8, and X9 are 12C; X3 is -12C(O)-; and X2 is ”C.

In another example, the compounds of formula I-B of this invention are represented in Table 1. It will be appreciated by those skilled in the art that the compounds of the t invention 2014/068713 may be represented in varying tautomeric forms.

Table 1 1-2 1-3 1-4 1-5 1-6 1-7 1-8 1-9 1-10 NH2 NH NH 13I O 2 /N A II0 2 /N O 4C 1.3C /l /N N / N /13C / /13 A x N C N/ N \ N \ N \ / H N—/( H H F F N—/< F <\ N N < N N < N N / \ / \ / F D F F DD D o o o K/N“W WD Dig/N D K/N”V b Do b o o b0 I-11 I-12 I-13 136; :9 /N N” 130‘ x / N \ / N H 1-14 Solid Forms Another aspect of the present invention provides a solid form of a compound of formula I- 3thNH2 0 / wherein the form is selected from the group consisting of Compound I-1° ethanol solvate, Compound I-1° e 1, nd I-1° hydrate 11, Compound I-1° anhydrous form A, Compound I-1° anhydrous form B, Compound I-1° anhydrous form C, Compound I-1° DMSO solvate, Compound I-1° DMAC e, Compound I-1° acetone solvate, and Compound I panol solvate. nd I-I° ethanol solvate In some aspects of the present inventions, the solid form is Compound I ethanol e. In another aspect of the present invention, the solid form is lline Compound I-1° ethanol solvate. In still other embodiments, crystalline Compound I-1° ethanol solvate has a Compound I-1 to ethanol ratio of about 1:0.72. In another aspect of the present invention, the crystalline Compound I-1° ethanol solvate is characterized by a weight loss of from about 5.76% in a temperature range of from about 166°C to about 219°C. In yet another aspect of the present invention, the crystalline Compound I-1° ethanol solvate is characterized by one or more peaks expressed in 2—theta :: 0.2 at about 17.2, 19.7, 23.8, 24.4, and 29.0 s in an X—Ray powder diffraction n obtained using Cu K alpha radiation. In other embodiments, the lline Compound I-1° l solvate is terized as having an X-ray powder diffraction pattern substantially the same as that shown in Figure 1a. In still other embodiments, the crystalline Compound I-1°ethanol solvate is characterized as having one or more peaks corresponding to 175.4 :: 0.3 ppm, 138.0 :: 0.3 ppm, 123.1 :: 0.3 ppm, 57.8 :: 0.3 ppm, 44.0 :: 0.3 ppm, and 19.5 :: 0.3 ppm in a C13 ssNMR spectrum. In yet another embodiment, the crystalline Compound I-1°ethanol solvate is characterized as having one or more peaks corresponding to —136.0 :: 0.3 ppm and —151.6 :: 0.3 ppm in an F19 ssNMR spectrum.

Compound I-I° hydrate I In some aspects of the present invention, the solid form is Compound I hydrate I. In another aspect of the present invention, the solid form is crystalline Compound I-1° hydrate I. In still other embodiments, the crystalline Compound I-1° hydrate I has a compound I-1 to H20 ratio of about 1:45. In yet another embodiment, crystalline Compound I-1° hydrate I is characterized by a weight loss of from about 14.56% in a temperature range of from about 25°C to about 100°C. In other embodiments, crystalline Compound I-1° e I is characterized by one or more peaks expressed in 2—theta :: 0.2 at about 6.5, 12.5, 13.7, 18.8, and 26.0 degrees in an X—Ray powder diffraction pattern ed using Cu K alpha radiation. In another embodiment, crystalline Compound I-1° hydrate I is characterized as having an X-ray powder diffraction pattern substantially the same as that shown in Figure 1b.

Compound I-I °hydrate II In some aspects of the present invention, the solid form is Compound I hydrate II. In another aspect of the present invention, the solid form is crystalline nd I-1° hydrate II. In other ments, crystalline Compound I-1° hydrate II is characterized by one or more peaks sed in 2-theta :: 0.2 at about 10.1, 11.3, 11.9, 20.2, and 25.1 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation. In still other embodiments, the crystalline nd Ihydrate II is characterized as having one or more peaks ponding to 177.0 :: 0.3 ppm, 158.2 0.3 ppm, 142.9 0.3 ppm, 85.1: 0.3 ppm, 58.9 0.3 ppm, and 31.9 0.3 ppm in a C13 ssNMR spectrum. In yet another ment, the lline Compound I-1°hydrate II is characterized as having one or more peaks ponding to —138.0 :: 0.3 ppm and —152.7 :: 0.3 ppm in an F19 ssNMR spectrum.

Compound I-I° anhydrousform A In one embodiment, the solid form is Compound I-1° anhydrous form A. In r embodiment, the solid form is crystalline Compound I-1° anhydrous form A. In still other embodiments, crystalline Compound I-1° anhydrous form A is characterized by a weight loss of from about 0.96 % in a temperature range of from about 25°C to about 265°C. In other embodiments, crystalline Compound I-1° anhydrous form A is characterized by one or more peaks expressed in 2—theta :: 0.2 at about 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X—Ray powder diffraction pattern obtained using Cu K alpha ion. In yet another embodiment, the lline nd I-1° anhydrous form A is characterized as having an X-ray powder diffraction pattern substantially the same as that shown in Figure 1c. In still other embodiments, the crystalline Compound Ianhydrous form A is characterized as having one or more peaks corresponding to 175.9 0.3 ppm, 138.9 0.3 ppm, 74.1 0.3 ppm, 42.8 0.3 ppm, and 31.5: 0.3 ppm in a c13 ssNMR spectrum. In yet another embodiment, the crystalline Compound I-1°anhydrous form A is characterized as having one or more peaks corresponding to —136.8 :: 0.3 ppm and —155.7 :: 0.3 ppm in an F19 ssNMR spectrum. One embodiment describes a process for preparing Compound I- 1°anhydrous form A comprising stirring a sion ning Compound I-1°ethanol solvate and a suitable organic solvent. In another embodiment, the suitable organic solvent is tetrahydrofuran.

Another aspect of the invention describes a process for preparing Compound I-1°anhydrous form A comprising stirring a suspension containing Compound I-1°amorphous, isopropanol, and water. In some embodiments, the suspension is heated to between about 65°C and about 80°C. In yet another embodiment, the suspension is heated to between about 70°C and about 75°C. In other embodiments, Compound I-1°anhydrous form A is characterized as a crystal form of Compound I-1 2014/068713 having a monoclinic crystal system, a P21/c centrosymmetric space group, and the following unit cell parameters: a = 15.29(3)A 0t = 900 b = 12.17(2)A [3: 107.22(3)O c = 14.48(3)A y = 90°.

Compound I-I° anhydrousform B As used herein, “anhydrous form B” refers to the THF solvate form of Compound I-1. In some embodiments, the solid form is Compound I anhydrous form B. In another embodiment, the solid form is crystalline Compound I-1° anhydrous form B. In yet another embodiment crystalline Compound I-1° anhydrous form B is characterized by a weight loss of from about 2.5 % in a temperature range of from about 25°C to about 175°C. In other embodiments, Compound I-1° anhydrous form B is characterized by one or more peaks expressed in 2-theta :: 0.2 at about 7.2, 8.3, 12.9, 19.5, and 26.6 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation. In still other embodiments, crystalline Compound I-1° anhydrous form B is characterized as haVing an X-ray powder diffraction pattern substantially the same as that shown in Figure Id. In still other embodiments, the crystalline Compound I-1°anhydrous form B is characterized as haVing one or more peaks corresponding to 173.4 :: 0.3 ppm, 164.5 :: 0.3 ppm, 133.5 :: 0.3 ppm, 130.8 :: 0.3 ppm, 67.7 :: 0.3 ppm, 45.3 :: 0.3 ppm, and 25.9 :: 0.3 ppm in a C13 ssNMR spectrum. In yet r embodiment, the crystalline Compound I-1°anhydrous form B is characterized as haVing one or more peaks corresponding to —138.0 :: 0.3 ppm and —153.5 :: 0.3 ppm in an F19 ssNMR spectrum.

Compound I-I° anhydrousform C In some embodiments, the solid form is Compound I ous form C.

In another embodiment, the solid form is crystalline Compound I-1° anhydrous form C. In other ments, crystalline Compound I-1° ous form C is characterized by one or more peaks expressed in 2—theta :: 0.2 at about 6.8, 13.4, 15.9, 30.9, and 32.9 degrees in an X—Ray powder ction pattern obtained using Cu K alpha radiation. In still other embodiments, crystalline Compound I-1° anhydrous form C is characterized as haVing an X-ray powder diffraction pattern substantially the same as that shown in Figure 1e. In still other embodiments, the crystalline Compound I-1°anhydrous form C is terized as haVing one or more peaks corresponding to 175.2 :: 0.3 ppm, 142.5 :: 0.3 ppm, 129.6 :: 0.3 ppm, 73.5: 0.3 ppm, 54.0 :: 0.3 ppm, and 46.7 :: 0.3 ppm in a C13 ssNMR spectrum. In yet another embodiment, the crystalline Compound I- drous form C is characterized as having one or more peaks corresponding to —131.2 :: 0.3 ppm and —150.7 :: 0.3 ppm in an F19 ssNMR spectrum.

Compound I-I °amorphous In some ments, the solid form is Compound I amorphous. In another embodiment, the solid form is crystalline Compound I-1° amorphous. In still other embodiments, the crystalline Compound orphous is characterized as having one or more peaks corresponding to 173.8 :: 0.3 ppm, 144.2 :: 0.3 ppm, 87.5 :: 0.3 ppm, 45.6 :: 0.3 ppm, and 29.5 :: 0.3 ppm in a C13 ssNMR spectrum. In yet another embodiment, the crystalline Compound I- 1-amorphous is characterized as having one or more peaks ponding to —137.7 :: 0.3 ppm and — 153.1 :: 0.3 ppm in an F19 ssNMR spectrum.

Compound I-I° DMSO e In one ment, the solid form is Compound I DMSO solvate. In another embodiment, the solid form is crystalline Compound I-1° DMSO solvate. In still other embodiments, the crystalline Compound I-1° DMSO solvate has a compound I-1° to DMSO ratio of about 1:1. In yet another embodiment, crystalline Compound I-1° DMSO solvate is terized by a weight loss of from about 12.44% in a temperature range of from about 146°C to about 156°C. In some embodiments, crystalline Compound I-1° DMSO solvate characterized by one or more peaks expressed in 2-theta :: 0.2 at about 8.9, 14.8, 16.5, 18.6, 20.9, 22.2, and 23.4 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha ion. In other ments, compound I- DMSO solvate is characterized as having an X-ray powder diffraction n substantially the same as that shown in Figure 1g.

Compound I-I° DMAC solvate In some embodiments, the solid form is Compound I DMAC solvate. In another embodiment, the solid form is crystalline Compound I-1° DMAC solvate. In other embodiments, the crystalline Compound I-1° DMAC solvate has a compound I-1 to DMAC ratio of about 1:13. In yet another embodiment, crystalline compound I-1° DMAC solvate is characterized by a weight loss of from about 17.76% in a temperature range of from about 85°C to about 100°C. In still other embodiments, compound I-1° DMAC solvate is characterized by one or more peaks expressed in 2— theta :: 0.2 at about 6.0, 15.5, 17.7, 18.1, 20.4, and 26.6 degrees in an X-Ray powder ction pattern obtained using Cu K alpha radiation. In some embodiments, compound I-1° DMAC solvate is characterized as having an X-ray powder diffraction pattern substantially the same as that shown in Figure 1h.

WO 85132 Compound I-I° acetone solvate In one or more emodiments, the solid form is Compound I acetone solvate. In another embodiment, the solid form is crystalline Compound I-1° acetone e. In yet another ment, the crystalline Compound I-1° e solvate has a compound I-1 to acetone ratio of about 1:0.44. In still other embodiment, Compound I-1° acetone solvate is characterized by a weight loss of from about 4.55% in a temperature range of from about 124°C to about 151°C. In some ments, Compound I-1° acetone solvate is characterized by one or more peaks expressed in 2— theta :: 0.2 at about 8.9, 15.5, 15.8, 16.7, 22.3, 25.7, and 29.0 degrees in an X-Ray powder ction n obtained using Cu K alpha radiation. In other embodiments, Compound I-1° acetone solvate is characterized as haVing an X-ray powder diffraction pattern substantially the same as that shown in Figure 1i.

Compound I-I° isopropanol solvate In one embodiment, the solid form is Compound I isopropanol solvate. In another embodiment, the solid form is crystalline nd I-1° isopropanol solvate. In still other ments, crystalline nd I-1° isopropanol solvate has a Compound I-1 to isopropanol ratio of about 1:0.35. In yet another embodiment, Compound I-1° isopropanol solvate is characterized by a weight loss of from about 3.76% in a temperature range of from about 136°C to about 180°C. In some embodiments, Compound I-1° isopropanol solvate is characterized by one or more peaks expressed in 2—theta :: 0.2 at about 6.9, 17.1, 17.2, 19.1, 19.6, 23.7, 24.4, and 28.9 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation. In another embodiment, Compound I-1° isopropanol solvate is characterized as haVing an X-ray powder diffraction pattern substantially the same as that shown in Figure 1j.

Formulation Some embodiments sed herein generally relate to a ition that can include an effective amount of Compound I-1, or a pharmaceutically acceptable salt f; and one or more excipients. Compound I-1 is believed to be an ATR inhibitor, and described in , which is hereby incorporated by reference in its entirety.

Compound I-1 and Form A can exist in free form or as a salt. Those salts that are pharmaceutically acceptable can be useful in administering Compound I-1 or Form A for medical purposes. Salts that are not pharmaceutically acceptable can be useful for manufacturing, isolating, purifying and/or separating stereoisomeric forms of Compound I-1, Form A and/or one or more intermediates thereof As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound, which are, within the scope of sound medical judgment, suitable for use in humans and lower animals without undue side effects, such as, toxicity, irritation, allergic se and the like, and are commensurate with a reasonable benefit/risk ratio. Various pharmaceutically acceptable salts can be used. For example, those salts sed in S. M. Berge et al., J. Pharmaceutical Sciences, 1977, 66, l-l9, which is hereby incorporated by reference. Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable nic and organic acids and bases. A salt of a compound described herein (for e, Compound I-l) can be ed in situ during the final isolation and purification of the compound.

As bed above, Compound I-1 can exist in different polymorphic forms (i.e., “solid ). Polymorphism is the ability of a compound to exist as more than one distinct crystalline or "polymorphic" species, wherein each species has a different ement of its molecules in the crystal lattice. Each distinct crystalline species is a “polymorph.” Each polymorph has the same chemical formula, however, can be display different physical property(ies) as a result of its ent arrangement in the crystal lattice. Polymorphs can be characterized by analytical methods such as X— ray powder diffraction (XRPD) pattern, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), melting point, and/or other ques known in the art.

Form A, described herein, can be in pure form or in a mixture with other materials.

Examples of other materials include, for example, other forms of Compound I-1 (such as amorphous forms, other polymorphic forms, es and hydrates); other diastereomers of Compound I-l; and/or other materials besides Compound LL Thus, in some embodiments, a composition can include an effective amount of pure Form A. As used herein, “pure” Form A is over 95% (w/w) (wherein w/w is weight of Form A/weight of Compound I-1 (wherein weight of Compound I-1 is weight of Form A + weight of all other forms of Compound I-1)), for e, over 98% (w/w), over 99% (w/w %), over 99.5% (w/w %), or over 99.9% (w/w %). In some embodiments, a composition can include an ive amount of Form A in an amount at least 95% (w/w %), at least 97% (w/w %) or at least 99% (w/w %) free of any other diastereomers of Compound I-1. In some embodiments, a composition can e an effective amount of Form A in an amount at least 95% (w/w %), at least 97% (w/w %) or at least 99% (w/w %) free of any other polymorphs and amorphous forms of Compound LL In some embodiments, a composition can include Form A with one or more other forms of Compound I-1. Other forms of Compound I-1 include, for example, hydrates, solvates, amorphous forms, other polymorphic forms, or combinations thereof.

WO 85132 In some embodiments, a composition can include an amount of Compound I-1 or Form A (or a pharmaceutically able salt of the aforementioned nds) in the range of a trace amount (0.1%) up to 100% (w/w %) relative to the total weight of the ition. In some embodiments, a composition can include less than about 50% of nd I-1 or Form A relative to the total weight of the composition (wherein the total weight includes the weight of Compound I-1 or Form A). For example, a composition can e an amount of Compound I-1 or Form A in a range selected from 0.1% — 0.5%, 0.1% — 1%, 0.1% — 2%, 0.1% — 5%, 0.1% — 10%, 0.1% — 20%, 0.1% — 30%, 0.1% — 40%, and 0.1% — <50% (w/w %) relative to the total weight of the composition (wherein the total weight includes the weight of Compound I-1 or Form A). In other embodiments, a composition can include equal to or greater than about 50% of Compound I-1 or Form A relative to the total weight of the composition (wherein the total weight includes the weight of Compound I-1 or Form A). For example, a composition can include at least 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, 99.5% or 99.9% (w/w) of Compound I-1 or Form A relative to the total weight of the ition (wherein the total weight includes the weight of Compound I-1 or Form A). In some embodiments, a composition can include an amount of Compound I-1 or Form A in the range of about 1 wt% to about 50 wt%; about 5 wt% to about 40 wt%, about 5 wt% to about 25 wt% or about wt% to about 15 wt% of Compound I-1 or Form A relative to the total weight of the composition (wherein the total weight includes the weight of Compound I-1 or Form A).

] As used herein, an “excipient” is used herein in its ordinary sense as understood by those skilled in the art, and includes one or more inert substances that are included in a composition to provide, t limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability etc., to the ition. Examples of excipients include fillers, binders, disintegrants, wetting agents, lubricants, glidants, humectants and absorbants.

In some embodiments, a ition can include Compound I-1 or Form A and one or more other components selected from one or more fillers, one or more binders, one or more disintegrants, one or more wetting agents and one or more lubricants. In some embodiments, a composition can include an amount of one or more fillers in the range of about 10 wt% to about 95 wt%; about 25 wt% to about 90 wt%; about 50 wt% to about 90 wt%; or about 70 wt% to about 90 wt% of the filler(s) by total weight of the ition (wherein the total weight includes the weight of one or more fillers). In some embodiments, a composition can include an amount of one or more lubricants in the range of about 0.1 wt% to about 10 wt%, about 0.5 wt% to about 7 wt%, or about 1 wt% to about 5 wt% of the lubricant(s) by total weight of the composition (wherein the total weight includes the weight of one or more lubricants). In some embodiments, a composition can include an amount of one or more disintegrants in the range of about 1 wt% to about 15 wt%, about 1 wt% to about 10 wt%, or about 1 wt% to about 7 wt% of the disintegrant(s) by total weight of the composition (wherein the total weight includes the weight of one or more disintegrants).

The wetting agents, binders, egrants, lubricants and fillers suitable for inclusion can be compatible with the ingredients of the compositions, for e, they do not substantially reduce the chemical stability of the active pharmaceutical ingredient(s).

The term “wetting agent” is used herein in its ordinary sense as understood by those skilled in the art, and es surfactants, such as non-ionic tants and anionic surfactants.

Wetting agents can enhance the solubility of the composition. Exemplary surfactants include sodium lauryl sulfate (SLS), polyoxyethylene sorbitan fatty acids (e. g., TWEENTM), sorbitan fatty acid esters (e.g., ), sodium dodecylbenzene sulfonate (SDBS), dioctyl sodium sulfosuccinate (Docusate), dioxycholic acid sodium salt (DOSS), sorbitan monostearate, sorbitan tristearate, sodium N- lauroylsarcosine, sodium oleate, sodium myristate, sodium stearate, sodium palmitate, Gelucire 44/ 14, ethylenediamine tetraacetic acid , Vitamin E d-alpha tocopheryl polyethylene glycol 1000 succinate , Lecithin, MW 677—692, Glutanic acid monosodium monohydrate, Labrasol, PEG 8 caprylic/capric glycerides, Transcutol, diethylene glycol monoethyl ether, Solutol HS-15, polyethylene /hydroxystearate, Taurocholic Acid, copolymers of polyoxypropylene and polyoxyethylene (e. g., poloxamers also known and commercially available under Pluronics®, such as, Pluronic® L61, Pluronic® F68, Pluronic® F108, and Pluronic® F127), saturated polyglycolized glycerides irs®), docusate sodium, polyoxyethylene an fatty acid esters, polyoxyethylene 20 stearyl ethers, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, pegylated hydrogenated castor oils, sorbitan esters of fatty acids, Vitamin E or tocol derivatives, vitamin E TPGS, eryl esters, lecithin, phospholipids and their tives, stearic acid, oleic acid, oleic alcohol, cetyl alcohol, mono and diglycerides, propylene glycol esters of fatty acids, glycerol esters of fatty acids, ethylene glycol palmitostearate, polyoxylglycerides, propylene glycol monocaprylate, propylene glycol monolaurate, yceryl oleate and any ations thereof. Sodium lauryl e is an anionic tant; and copolymers of ypropylene and polyoxyethylene are non-ionic tants. Specific examples of copolymers of polyoxypropylene and polyoxyethylene include poloxamers, such as a poloxamer with a polyoxypropylene molecular mass of 1,800 g/mol and a 80% polyoxyethylene content (e. g., poloxamer 188).

The term “binder” is used herein in its ordinary sense as understood by those skilled in the art, and includes agents used while making granules of the active ingredient (for example, Compound I-1 or Form A), wherein a binder holds the active ingredient together with one or more inactive agents. Exemplary binders include polyvinyl pyrrolidones (PVPs), pregelatinized starch, starch, microcrystalline cellulose, modifled cellulose (e. g., hydroxyl propyl methyl cellulose (HPMC), ypropyl cellulose (HPC) and hydroxy ethyl ose (HEC)), and any combination f. PVP’s are commonly characterized by the “K-value,” which is a measurement of the polymeric composition's viscosity. PVPs can be commercially sed (e. g., Tokyo Chemical Industry Co., Ltd.) under the trade name of Povidone® K12, Povidone® K17, Povidone® K25, Povidone® K30, Povidone® K60, and Povidone® K90. Specific examples of PVPs include soluble spray dried PVP. PVPs can have an e molecular weight of 3,000 daltons to 4,000 daltons, such as Povidone® K12 having an average lar weight of 4,000 daltons. PVP can be used in either a wet or a dry state.

The term “filler” (or “diluent”) is used herein in its ordinary sense as understood by those skilled in the art, and includes microcrystalline celluloses (e.g., Avicel® PH 101), lactoses, ols, celluoses, calcium phosphates, starches, sugars (e.g., mannitol, sucrose, or the like), dextrose, maltodextrin, sorbitol, xylitol, powdered cellulose, silicifled microcrystalline cellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, methylhydroxyethylcellulose, pregelatinized starch, dibasic calcium phosphate, calcium sulfate, calcium carbonate and any combination thereof.

Specific examples of fillers include microcrystalline celluloses and lactoses. Specific examples of microcrystalline celluloses include commercially available Avicel® series, such as microcrystalline celluloses having a particle size of 200 mesh over 70% and a particle size of 65 mesh less than 10% (e.g., Avicel® PH 101). A specific e of a lactose is lactose monohydrate.

The term “disintegrant” is used herein in its ordinary sense as understood by those skilled in the art, and can enhance the dispersal of a composition. Examples of disintegrants include croscarmellose , starch (e.g., corn starch, potato starch), sodium starch ate, crospovidone, microcrystalline ose, sodium alginate, calcium alginate, alginic acid, pregelatinized starch, cellulose and its derivatives, carboxymethylcellulose calcium, carboxymethylcellulose sodium, soy polysaccharide, guar gum, ion exchange , an effervescent system based on food acids and an alkaline carbonate ent, sodium onate and any combinations thereof. Specific examples of disintegrants include croscarmellose sodium (e. g., Ac— Di-Sol®) and sodium starch glycolate.

The term “lubricant” is used herein in its ordinary sense as understood by those skilled in the art, and can e the compression and ejection of a composition, e. g., through a die press.

Exemplary lubricants include magnesium stearate, stearic acid (stearin), hydrogenated oils, sodium stearyl te, sodium lauryl sulfate, talc, fatty acid, calcium stearate, sodium stearate, glyceryl monostearate, fatty alcohol, fatty acid ester, glyceryl behenate, mineral oil, vegetable oil, leucine, sodium te and any combination thereof. A ic example of a lubricant is sodium stearyl fumarate.

Those skilled in the art understand that a specific compound described as a wetting agent, binder, filler, disintegrant and lubricant can serve one or more purpose. For example, microcrystalline cellulose can be used as a egrant and filler.

] In some embodiments, a composition can include an amount of Compound 1-1 or Form A in the range of about 5 wt% to about 50 wt% by the total weight of the composition; and an amount of one or more fillers in the range of about 10 wt% to about 90 wt% by the total weight of the composition. In other embodiments, a composition can include an amount of Compound 1-1 or Form A in the range of about 5 wt% to about 50 wt% by the total weight of the composition; an amount of one or more fillers in the range of about 10 wt% to about 90 wt% by the total weight of the composition; and an amount of one or more egrants in the range of about 1 wt% to about 15 wt% by the total weight of the composition. In still other ments, a composition can include an amount of Compound 1-1 or Form A in the range of about 5 wt% to about 50 wt% by the total weight of the composition; an amount of one or more fillers in the range of about 10 wt% to about 90 wt% by the total weight of the ition; an amount of one or more disintegrants in the range of about 1 wt% to about 15 wt% by the total weight of the composition; and an amount of one or more lubricants in the range of about 0.1 wt% to about 10 wt% by the total weight of the composition.

In some embodiments, a ition can include an amount of Compound 1-1 or Form A in the range of about 5 wt% to about 20 wt% by the total weight of the composition; an amount of one or more lubricants in the range of about 1 wt% to about 5 wt% by the total weight of the composition; an amount of one or more egrants in the range of about 1 wt% to about 10 wt% by the total weight of the composition; and an amount of one or more fillers in the range of about 70 wt% to about 90 wt% by the total weight of the composition. In other embodiments, a composition can include an amount of Compound 1-1 or Form A in the range of about 5 wt% to about 15 wt% by the total weight of the composition; an amount of one or more lubricants in the range of about 1 wt% to about 5 wt% by the total weight of the composition; an amount of one or more disintegrants in the range of about 1 wt% to about 5 wt% by the total weight of the composition; and an amount of one or more fillers in the range of about 70 wt% to about 90 wt% by the total weight of the composition.

In some embodiments, a composition can include an amount of Compound 1-1 or Form A of about 10 wt% by the total weight of the composition, an amount of lactose monohydrate of about 28 wt% by the total weight of the composition, an amount of AVicel PH-101 crystalline cellulose) of about 55 wt% by the total weight of the composition, an amount of Ac—Di—Sol (croscarmellose sodium) of about 5 wt% by the total weight of the composition, and an amount of sodium stearyl fumarate of about 3 wt% by the total weight of the composition.

In some ments, a composition can further e one or more glidants (or “flow aids”). A glidant enhances the flow properties of a composition by reducing interparticle friction and cohesion. Exemplary glidants include colloidal silicon dioxide, talc, and any combination f. A specific example of glidant is amorphous, colloidal silicon dioxide having an average le size in 0.2 — 0.3 microns, such as Cab—O—Sil® MSP. The amount of a glidant can vary. For example, the amount of glidant(s) can be in the range of about 0.1 wt% to about 3 wt%, or about 0.1 wt% to about 1 wt% by total weight of the composition (wherein the total weight es the weight of one or more glidants).

In some embodiments, a composition bed herein can further include a g.

] In some embodiments, a composition described herein can be in a solid dosage form, for example, a tablet.

Some embodiments described herein relate to a method of preparing a composition described herein. In some embodiments, a method can include providing a mixture that includes Compound 1-1 or Form A and one or more fillers to form a ition. In other embodiments, a method can include providing a mixture that includes Compound 1-1 or Form A, a lubricant, a egrant, and a filler to form a composition. Examples, including specific examples, of lubricants, disintegrants, and fillers are each and independently bed herein.

In some embodiments, a method can include combining Compound 1-1 or Form A and one or more first excipients to form a mixture; and combining the mixture (that includes Compound 1-1 or Form A and one or more first excipients) with one or more second excipients. In some embodiments, the first excipients can include one or more of the following: one or more fillers, one or more disintegrants, and one or more lubricants. In some embodiments, the second excipients can include one or more of the following: one or more disintegrants and one or more ants.

In other ments, a method of preparing a composition described herein can include: i) combining Compound 1-1 or Form A with one or more first excipients that can include one or more fillers, one or more disintegrants and one or more lubricants, and ii) combining the mixture from i) with one or more second excipients that can include one more disintegrants and one or more lubricants to form a composition. In some embodiments, the one or more first excipients can include an amount of one or more fillers in the range of about 70 wt% to about 90 wt%, an amount of one or more disintegrants in the range of about 1 wt% to about 15 wt%, and an amount of one or more lubricants in the range of about 1 wt% to about 5 wt% each by the total weight of the ition, and the second excipients can include an amount of one or more lubricants in the range of about 0.5 wt% to about 5 wt% and an amount of one or more disintegrants in the range of about 0.5 wt% to about 5 wt% each by the total weight of the composition.

WO 85132 In some embodiments, a method of preparing a composition described herein can include: i) providing granules of Compound 1-1 or Form A by combining nd 1-1 or Form A with first ents that may include one or more fillers, one or more disintegrants, and one or more lubricants; and ii) mixing the granules of Compound 1-1 or Form A obtained from i) with second ents that may include one or more disintegrants and one or more lubricants and optionally one or more flllers to form a composition. In some embodiments, the first excipients can include an amount of one or more fillers in the range of about 70 wt% to about 90 wt%, an amount of one or more disintegrants in the range of about 0.5 wt% to about 5 wt%, and an amount of a first lubricant in the range of about 1% to about 5% each by the total weight of the composition; and the second ents can include an amount of one or more second lubricants in the range of about 0.5 wt% to about 5 wt% and an amount of one or more disintegrants in the range of about 0.5 wt% to about 5 wt% each by the total weight of the composition. Examples, including specific examples, of suitable ants, disintegrants, and fillers are described herein.

In some embodiments, a method of preparing a composition described herein can include passing Compound 1-1 or Form A through a sieve; mixing granules of Compound 1-1 or Form A with one or more flllers, one or more disintegrants, and one or more lubricants; and blending the resulting granules with one or more disintegrants and one or more lubricants.

In some embodiments, a method of preparing a composition described herein can e compressing granules that include Compound 1-1 or Form A through a tablet compression machine to form a tablet that includes Compound 1-1 or Form A.

In some embodiments, a tablet that can include Compound 1-1 or Form A (for example, the tablets obtained after tablet compression) can be film coated.

The itions described herein may further include one or more pharmaceutically acceptable carriers other than those described previously. As used herein, “pharmaceutically acceptable” means being inert without unduly inhibiting the ical activity of the compounds.

The pharmaceutically acceptable carriers should be biocompatible, e.g., non—toxic, non— inflammatory, non—immunogenic or devoid of other undesired reactions or side—effects upon the administration to a t. r, standard pharmaceutical ation techniques can be employed for integrating the aforementioned one or more ceutically acceptable carriers.

Some examples of materials which can serve as pharmaceutically able carriers include, but are not limited to, ion exchangers; alumina; aluminum stearate; in; serum proteins (such as human serum albumin); buffer substances (such as phosphates or glycine); partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, or zinc salts); WO 85132 colloidal silica; magnesium trisilicate; polyacrylates; waxes; polyethylene-polyoxypropylene—block polymers; methylcellulose; ypropyl methylcellulose; wool fat; sugars such as e; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols; such a propylene glycol or hylene ; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; other xic compatible lubricants; coloring agents; releasing agents; sweetening; flavoring agents; ing agents; preservatives; sorbents and antioxidants can also be present in the composition, according to the judgment of the formulator.

Some embodiments bed herein relate to a method of inhibiting or reducing the activity ofATR in a subject that can include stering to the subject a composition described herein that contains an effective amount of Compound 1-1 or Form A, or a ceutically acceptable salt the aforementioned compounds.

] Other embodiments described herein relate to a method of treating cancer in a subject that can include administering to the subject a ition described herein that contains an effective amount of Compound 1-1 or Form A, or a pharmaceutically acceptable salt the aforementioned compounds.

Yet still other embodiments described herein relate to an use of a composition described herein that contains an effective amount of nd 1 or Form A, or a pharmaceutically acceptable salt the aforementioned compounds, in the manufacture of a medicament for treating cancer.

In some embodiments, substantially all by weight of Compound 1-1 in a composition described herein can be Form A.

In some embodiments, at least 90% by weight of Compound 1-1 in a composition described herein can be Form A.

In some embodiments, at least 95% by weight of Compound 1-1 in a composition bed herein can be Form A.

In some embodiments, at least 98% by weight of Compound 1-1 in a composition described herein can be Form A.

In some ments, at least 99% by weight of Compound 1-1 in a composition described herein can be Form A.

The compositions described herein can be administered to humans and other animals WO 85132 orally, rectally, parenterally, intracistemally, intravaginally, eritoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray, or the like. The term "parenteral" as used herein includes, but is not limited to, subcutaneous, intravenous, intramuscular, intra— articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In some embodiments, a composition described herein can be administered orally, intraperitoneally and/or intravenously.

Any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous sions or solutions. In the case of tablets, suitable carriers used include, but are not d to, e and corn starch. Lubricating agents, such as magnesium stearate, and/or wetting agents can be added. When aqueous suspensions are used, the active ingredient can be combined with emulsifying and/or suspending agents. If desired, sweetening, flavoring, coloring agents and/or perfuming agents can be included.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.

In addition to the active compound, the liquid dosage forms may contain inert excipients, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl l, benzyl te, propylene glycol, 1,3— butylene glycol, dimethylformamide, oils (such as, cottonseed, groundnut, corn, germ, olive, , and sesame oils), glycerol, ydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

] Solid dosage forms for oral administration include capsules (for example, soft and hard- filled gelatin capsules), tablets, pills, s, and granules. In such solid dosage forms, the active compound can be mixed with at least one inert, ceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers such as starches, lactose, milk sugar, sucrose, glucose, ol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar, calcium ate, potato or a starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as nary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) ents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form can also include a buffering agent.

The solid dosage forms of s, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other gs known in the pharmaceutical formulating art. They may optionally n opacifying agents and can also be of a composition that can release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active nd(s) can be in a microencapsulated form with one or more excipients.

Sterile injectable forms may be aqueous or oleaginous suspension. Injectable preparations may be formulated ing to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable ation may be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally able diluent or solvent, for example, as a solution in propylene . Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils can be employed as a solvent or ding . For this purpose any bland f1xed oil can be employed including synthetic mono— or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of inj ectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain l diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions.

Inj ectable formulations can be sterilized, for example, by filtration through a bacterialretaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

Dosage forms for topical or transdermal administration include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and patches. The active component can be d under sterile conditions with a pharmaceutically acceptable carrier, and any preservatives and/or buffers may be included. Ophthalmic formulation, ps, and eye drops can be formulated. Such dosage forms can be made by ving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be lled by either providing a rate controlling membrane or by sing the compound in a polymer matrix or gel.

] Alternatively, the active compounds and pharmaceutically acceptable compositions thereof may also be administered by nasal aerosol or inhalation. Such compositions are ed according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in , employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.

] Surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers can be included in a solid, liquid and other dosage forms described herein.

The compositions described herein can be formulated in an unit dosage form. The term “unit dosage form” refers to physically discrete units suitable as unitary dosage for subjects undergoing treatment, with each unit containing a predetermined quantity of active al calculated to e the d eutic effect, optionally in association with a le pharmaceutical carrier. The unit dosage form can be for a single daily dose or one of multiple daily doses (e. g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage form can be the same or ent for each dose. The amount of the active compound in a unit dosage form will vary depending upon, for example, the host treated, and the particular mode of administration, for example, from 0.01 mg/kg body weight/dose to 100 mg/kg body weight/dose.

In some embodiments, a compositions described herein can be in the form of a solid dosage form. In some embodiments, a composition described herein can be in the form of a tablet.

In still other embodiments, the composition may be in the form of a 100 mg tablet, or a 500 mg tablet.

It will be appreciated that the amount of the active compound (for example, Compound I- 1 or Form A) required for use in treatment will vary not only with the particular compound ed but also with the route of administration, the nature of the condition for which treatment is required and the age and condition of the subject and will be ultimately at the discretion of the attendant physician or veterinarian. In general, however, a suitable dose will be in the range of from about 0.1 to about 100 mg/kg of body weight per dose, for example, in the range of 0.5 to 50 mg/kg/dose, or, for example, in the range of l to 10 mg/kg/dose.

In some embodiments, a composition described herein can be administered in an amount in the range of about 5 mg to about 100 mg of Compound 1-1 or Form A, or a pharmaceutically acceptable salt the entioned compounds, per dose.

In some ments, a composition described herein can be stered: a) in an amount of about 5 mg Compound 1-1 or Form A, or a pharmaceutically acceptable salt the aforementioned compounds, per dose; b) in an amount of about 10 mg Compound 1-1 or Form A, or a pharmaceutically acceptable salt the aforementioned compounds, per dose; c) in an amount of about 20 mg Compound 1-1 or Form A, or a pharmaceutically acceptable salt the entioned compounds, per dose; d) in an amount of about 30 mg Compound 1-1 or Form A, or a pharmaceutically acceptable salt the aforementioned compounds, per dose; e) in an amount of about 50 mg Compound 1-1 or Form A, or a pharmaceutically able salt the aforementioned compounds, per dose; f) in an amount of about 60 mg Compound 1-1 or Form A, or a pharmaceutically acceptable salt the aforementioned compounds, per dose; g) in an amount of about 80 mg Compound 1-1 or Form A, or a pharmaceutically able salt the aforementioned compounds, per dose; or h) in an amount of about 100 mg Compound 1-1 or Form A, or a pharmaceutically acceptable salt the aforementioned compounds, per dose.

] In some embodiments, a composition bed herein can be administered in a fasted state (for example, the subject has not eaten food or liquids, except for water, for at least 8 hours). In other embodiments, a composition described herein can be administered in a fed state (for example, with food or within 1 hour of eating food).

Comp_ound Uses ] One aspect of this invention provides compounds or compositions that are inhibitors of ATR kinase, and thus are useful for treating or lessening the severity of a disease, condition, or disorder in a subject or patient where ATR is implicated in the disease, condition, or disorder.

] Another aspect of this invention provides compounds or itions that are useful for the treatment of diseases, disorders, and conditions characterized by ive or abnormal cell proliferation. Such diseases include a proliferative or hyperproliferative disease. Examples of proliferative and hyperproliferative diseases include, without limitation, cancer and myeloproliferative disorders.

In some embodiments, said compounds are ed from nd 1-1 or Form A. In other embodiments, said compositions include compound 1-1 or Form A. The term “cancer” includes, but is not limited to the following cancers. Oral: buccal cavity, lip, tongue, mouth, pharynx; Cardiac: sarcoma (angiosarcoma, arcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; mg: all cell, bronchogenic carcinoma (squamous cell or epidermoid, undifferentiated small cell, erentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous oma, mesothelioma; intestinal: esophagus (squamous cell carcinoma, larynx, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, WO 85132 2014/068713 carcinoid tumors, vipoma), small bowel or small intestines (adenocarcinoma, lymphoma, carcinoid tumors, Karposi's sarcoma, leiomyoma, hemangioma, lipoma, neurof1broma, f1broma), large bowel or large intestines (adenocarcinoma, tubular a, villous adenoma, hamartoma, oma), colon, colon—rectum, colorectal; rectum, Genitourinam tract: kidney (adenocarcinoma, Wilm's tumor [nephroblastoma], lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, ma, embryonal oma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell oma, f1broma, f1broadenoma, adenomatoid tumors, lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma, biliary passages; Bone: osteogenic a (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, osarcoma, Ewing's sarcoma, malignant lymphoma ulum cell a), multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell ; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma [pinealoma], glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital ), spinal cord neurof1broma, meningioma, glioma, sarcoma); logical/Female: uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervical sia), ovaries (ovarian carcinoma [serous enocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosa-thecal cell , Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial oma, adenocarcinoma, f1brosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma), breast; Hematologic: blood (myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic cytic ia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignant lymphoma] hairy cell; lymphoid disorders; &: ant melanoma, basal cell carcinoma, squamous cell carcinoma, Karposi's sarcoma, keratoacanthoma, moles dysplastic nevi, lipoma, a, dermatof1broma, keloids, psoriasis, Thyroid gland: papillary thyroid carcinoma, follicular thyroid carcinoma, undifferentiated thyroid cancer, ary thyroid carcinoma, multiple endocrine neoplasia type 2A, multiple endocrine neoplasia type 2B, familial medullary thyroid cancer, pheochromocytoma, paraganglioma; and Adrenal glands: neuroblastoma.

In some embodiments, the cancer is selected from a cancer of the lung or the pancreas. In other embodiments, the cancer is ed from lung cancer, head and neck cancer, pancreatic cancer, gastric cancer, or brain cancer. In yet other embodiments, the cancer is selected from non-small cell lung cancer, small cell lung cancer, pancreatic cancer, biliary tract cancer, head and neck cancer, r cancer, colorectal cancer, glioblastoma, esophageal cancer, breast , hepatocellular oma, or ovarian cancer.

In some embodiments, the cancer is lung cancer. In other embodiments, the lung cancer is all cell lung cancer or small cell lung cancer. In r embodiment, the cancer is non-small cell lung . In yet another embodiment, the non-small cell lung cancer is squamous non-small cell lung .

Thus, the term "cancerous cell" as provided herein, includes a cell afflicted by any one of the above—identified conditions. In some embodiments, the cancer is selected from colorectal, thyroid, or breast cancer. In other embodiments, the cancer is triple negative breast cancer.

The term “myeloproliferative disorders”, includes disorders such as polycythemia vera, ocythemia, myeloid metaplasia with myelofibrosis, hypereosinophilic syndrome, juvenile myelomonocytic leukemia, systemic mast cell disease, and hematopoietic disorders, in particular, acute—myelogenous leukemia (AML), chronic—myelogenous leukemia (CML), acute—promyelocytic leukemia (APL), and acute cytic leukemia (ALL).

Combination ies Another aspect of this invention is directed towards a method of treating cancer in a subject in need thereof, comprising administration of a compound or composition of this invention or a pharmaceutically able salt thereof, and an additional therapeutic agent. In some embodiments, said method comprises the sequential or co—administration of the compound or composition (or a pharmaceutically acceptable salt thereof), and the onal therapeutic agent.

As used herein, the term “in combination” or “co-administration” can be used interchangeably to refer to the use of more than one therapy (e. g., one or more eutic agents).

The use of the term does not restrict the order in which therapies (e. g., eutic agents) are administered to a subject or the dosing schedule of each therapeutic agent.

In some embodiments, said additional therapeutic agent is an ancer agent. In other embodiments, said additional therapeutic agent is a DNA-damaging agent. In yet other embodiments, said additional therapeutic agent is selected from radiation therapy, chemotherapy, or other agents typically used in combination with radiation therapy or chemotherapy, such as radiosensitizers and chemosensitizers. In yet other embodiments, said onal therapeutic agent is ionizing radiation.

As would be known by one of skill in the art, radiosensitizers are agents that can be used in ation with radiation therapy. Radiosensitizers work in various different ways, including, but not limited to, making cancer cells more sensitive to radiation therapy, working in synergy with radiation therapy to e an improved synergistic effect, acting additively with radiation therapy, or protecting surrounding healthy cells from damage caused by ion therapy. Likewise chemosensitizers are agents that can be used in combination with chemotherapy. Similarly, chemosensitizers work in various different ways, including, but not limited to, making cancer cells more sensitive to chemotherapy, working in synergy with chemotherapy to provide an improved istic effect, acting additively to chemotherapy, or protecting surrounding healthy cells from damage caused by chemotherapy.

Examples of DNA-damaging agents that may be used in combination with compounds or compositions of this invention e, but are not limited to Platinating agents, such as Cisplatin, Carboplatin, Nedaplatin, latin and other derivatives; Topo I inhibitors, such as Topotecan, irinotecan/SN3 8, rubitecan and other derivatives; Antimetabolites, such as Folic family (Methotrexate, exed and ves); Purine antagonists and Pyrimidine antagonists (Thioguanine, Fludarabine, Cladribine, Cytarabine, Gemcitabine, 6-Mercaptopurine, 5-Fluorouracil (SFU) and relatives); Alkylating agents, such as Nitrogen mustards (Cyclophosphamide, Melphalan, Chlorambucil, mechlorethamine, mide and relatives); nitrosoureas (eg Carmustine); Triazenes (Dacarbazine, lomide); Alkyl sulphonates (eg Busulfan); Procarbazine and Aziridines; Antibiotics, such as Hydroxyurea, Anthracyclines (doxorubicin, daunorubicin, epirubicin and other derivatives); cenediones (Mitoxantrone and relatives); Streptomyces family (Bleomycin, Mitomycin C, actinomycin); and Ultraviolet light.

In some embodiments, the additional eutic agent is ionizing radiation. In other embodiments, the additional therapeutic agent is Cisplatin or Carboplatin. In yet other ments, the additional therapeutic agent is Etoposide. In yet other embodiments, the additional therapeutic agent is Temozolomide. In still other embodiments, the additional therapeutic agent is ecan/SN3 8.

In certain embodiments, the additional therapeutic agent is selected from one or more of the following: Cisplatin, Carboplatin, irinotecan/SN3 8, gemcitabine, Etoposide, lomide, or ionizing radiation.

Other therapies or anticancer agents that may be used in combination with the inventive compounds and compositions of the present invention include surgery, herapy (in but a few examples, gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive es, to name a few), endocrine therapy, biologic response modifiers (interferons, interleukins, and tumor necrosis factor (TNF) to name a few), hyperthermia and cryotherapy, agents to attenuate any adverse effects (e.g., antiemetics), and other approved chemotherapeutic drugs, including, but not limited to, the DNA damaging agents listed herein, e poisons (Vinblastine, Vincristine, Vinorelbine, Paclitaxel), podophyllotoxins (Etoposide, Irinotecan, Topotecan), nitrosoureas (Carmustine, Lomustine), inorganic ions (Cisplatin, Carboplatin), enzymes (Asparaginase), and hormones (Tamoxifen, lide, ide, and Megestrol), GleevecTM, ycin, dexamethasone, and cyclophosphamide.

A compound or composition of the instant invention may also be useful for treating cancer in combination with any of the following therapeutic : abarelix (Plenaxis depot®); aldesleukin (Prokine®); Aldesleukin ukin®); Alemtuzumabb (Campath®); alitretinoin (Panretin®); allopurinol rim®); altretamine (Hexalen®); amifostine (Ethyol®); anastrozole dex®); arsenic trioxide (Trisenox®); asparaginase (Elspar®); azacitidine (Vidaza®); zimab (Avastin®); bexarotene capsules (Targretin®); bexarotene gel etin®); bleomycin xane®); bortezomib (Velcade®); busulfan intravenous (Busulfex®); busulfan oral (Myleran®); calusterone (Methosarb®); capecitabine (Xeloda®); carboplatin (Paraplatin®); carmustine , BiCNU®); carmustine (Gliadel®); carmustine with Polifeprosan 20 Implant (Gliadel Wafer®); celecoxib (Celebrex®); cetuximab (Erbitux®); chlorambucil (Leukeran®); cisplatin (Platinol®); cladribine (Leustatin®, 2-CdA®); clofarabine (Clolar®); hosphamide (Cytoxan®, Neosar®); cyclophosphamide (Cytoxan Inj ection®); cyclophosphamide (Cytoxan Tablet®); cytarabine (Cytosar-U®); bine liposomal (DepoCyt®); dacarbazine (DTIC— Dome®); dactinomycin, actinomycin D (Cosmegen®); Darbepoetin alfa (Aranesp®); daunorubicin liposomal (DanuoXome®); daunorubicin, daunomycin rubicin®); daunorubicin, daunomycin (Cerubidine®); Denileukin diftitox (Ontak®); oxane (Zinecard®); docetaxel (Taxotere®); doxorubicin mycin PFS®); doxorubicin (Adriamycin®, ); doxorubicin (Adriamycin PFS Injection®); doxorubicin liposomal (Doxil®); dromostanolone propionate (dromostanolone®); dromostanolone nate (masterone injection®); Elliott's B Solution (Elliott's B Solution®); epirubicin (Ellence®); n alfa (epogen®); erlotinib (Tarceva®); estramustine (Emcyt®); etoposide ate (Etopophos®); etoposide, VP—l6 (Vepesid®); exemestane (Aromasin®); Filgrastim (Neupogen®); floxuridine (intraarterial) (FUDR®); fludarabine (Fludara®); fluorouracil, —FU (Adrucil®); fulvestrant (Faslodex®); gef1tinib (Iressa®); gemcitabine (Gemzar®); gemtuzumab ozogamicin arg®); goserelin acetate (Zoladex Implant®); goserelin acetate (Zoladex®); histrelin acetate (Histrelin implant®); hydroxyurea (Hydrea®); momab Tiuxetan (Zevalin®); idarubicin (Idamycin®); ifosfamide (IFEX®); imatinib mesylate (Gleevec®); eron alfa 2a (Roferon A®); Interferon alfa-2b (Intron A®); irinotecan (Camptosar®); lenalidomide (Revlimid®); letrozole (Femara®); leucovorin (Wellcovorin®, Leucovorin®); Leuprolide Acetate (Eligard®); levamisole (Ergamisol®); lomustine, CCNU ®); ethamine, nitrogen mustard (Mustargen®); megestrol acetate (Megace®); melphalan, L-PAM (Alkeran®); mercaptopurine, 6-MP (Purinethol®); mesna (Mesnex®); mesna (Mesnex ; methotrexate trexate®); methoxsalen (Uvadex®); mitomycin C (Mutamycin®); mitotane (Lysodren®); mitoxantrone (Novantrone®); nandrolone phenpropionate (Durabolin-50®); nelarabine (Arranon®); Nofetumomab ma®); Oprelvekin (Neumega®); oxaliplatin (Eloxatin®); paclitaxel (Paxene®); paclitaxel (Taxol®); paclitaxel n-bound particles (Abraxane®); palifermin (Kepivance®); pamidronate a®); pegademase (Adagen (Pegademase Bovine)®); argase (Oncaspar®); Pegfilgrastim (Neulasta®); pemetrexed disodium (Alimta®); pentostatin (Nipent®); pipobroman (Vercyte®); ycin, mithramycin (Mithracin®); porfimer sodium (Photofrin®); procarbazine (Matulane®); quinacrine (Atabrine®); Rasburicase (Elitek®); Rituximab (Rituxan®); sargramostim (Leukine®); Sargramostim (Prokine®); sorafenib ar®); streptozocin (Zanosar®); sunitinib maleate (Sutent®); talc (Sclerosol®); tamoxifen (Nolvadex®); temozolomide (Temodar®); teniposide, VM—26 (Vumon®); testolactone (Teslac®); thioguanine, 6-TG (Thioguanine®); thiotepa (Thioplex®); topotecan (Hycamtin®); toremifene (Fareston®); Tositumomab (Bexxar®); Tositumomab/I-l3l momab (Bexxar®); Trastuzumab (Herceptin®); tretinoin, ATRA (Vesanoid®); Uracil d (Uracil Mustard es®); valrubicin (Valstar®); vinblastine (Velban®); vincristine (Oncovin®); vinorelbine (Navelbine®); zoledronate (Zometa®) and vorinostat (Zolinza®).

For a comprehensive discussion of updated cancer therapies see, http://www.nci.nih.gov/, a list of the FDA approved oncology drugs at http://wwwfda.gov/cder/cancer/druglistframe.htm, and The Merck Manual, Seventeenth Ed. 1999, the entire ts of which are hereby incorporated by reference.

] Another embodiment provides administering a compound or ition of this invention with an onal therapeutic agent that inhibits or modulates a base excision repair protein. In some embodiments, the base excision repair protein is selected from UNG, SMUGl, MBD4, TDG, OGGl, MYH,NTH1, MPG, NEILl, NEIL2, NEIL3 (DNA glycosylases); APEl, APEXZ (AP endonucleases); LIGl, LIG3 (DNA ligases I and III); XRCCl (LIG3 accessory); PNK, PNKP ucleotide kinase and phosphatase); PARPl, PARP2 (Poly(ADP-Ribose) Polymerases); PolB, PolG (polymerases); FENl (endonuclease) or Aprataxin. In other ments, the base excision repair protein is selected from PARP l, PARPZ, or PolB. In yet other embodiments, the base excision repair protein is selected from PARPl or PARPZ. In some embodiments, the agent is selected from Olaparib (also known as AZD2281 or 9436), ib (also known as BSI-201 or SAR240550), Veliparib (also known as ABT-888), Rucaparib (also known as PF—01367338), CEP— 9722, INO—lOOl, MK—4827, E7016, BMN673, or AZD2461.

Methods of Treatment One aspect of the invention relates to a method of inhibiting ATR kinase activity in a patient, which method comprises stering to the patient a compound described herein, or a composition comprising said compound. In some embodiments, said method is used to treat or prevent a condition selected from proliferative and hyperproliferative diseases, such as cancer.

In some embodiments, the cancer is selected from the s described herein. In some ments, said cancer is lung cancer, head and neck cancer, pancreatic cancer, gastric cancer, or brain cancer. In other ments, the cancer is ed from a cancer of the lung or the pancreas.

In yet other embodiments, the cancer is selected from non-small cell lung cancer, small cell lung cancer, pancreatic cancer, biliary tract cancer, head and neck cancer, bladder cancer, colorectal cancer, glioblastoma, esophageal cancer, breast cancer, hepatocellular carcinoma, or ovarian cancer.

In some embodiments, the lung cancer is small cell lung cancer and the onal therapeutic agents are cisplatin and etoposide. In other es, the lung cancer is non-small cell lung cancer and the additional therapeutic agents are gemcitabine and cisplatin. In yet other embodiments, the all cell lung cancer is squamous non-small cell lung cancer. In another embodiment, the cancer is breast cancer and the onal eutic agent is cisplatin. In other embodiments, the cancer is triple negative breast cancer.

In certain embodiments, an "effective amount" of the compound or pharmaceutically acceptable composition is that amount effective in order to treat said disease. The compounds and compositions, according to the method of the present invention, may be administered using any amount and any route of administration effective for treating or lessening the severity of said disease.

One aspect provides a method for inhibiting ATR in a t comprising administering a nd or composition as described herein. Another embodiment provides a method of treating cancer comprising administering to a t a compound or composition described herein, n the variables are as defined .

Another ment provides methods for treating pancreatic cancer by administering a compound or composition described herein in combination with another known pancreatic cancer treatment. One aspect of the invention includes administering a nd or composition bed herein in combination with gemcitabine. In some ments, the pancreatic cancer comprises one of the following cell lines: PSN—l, MiaPaCa-2 or . According to another aspect, the cancer comprises one of the following primary tumor lines: Panc-M or MRCS.

Another aspect of the invention includes administering a compound or composition described herein in combination with radiation therapy. Yet another aspect provides a method of abolishing radiation-induced G2/M checkpoint by stering a compound or composition described herein in combination with radiation treatment.

Another aspect provides a method of treating pancreatic cancer by administering to pancreatic cancer cells a compound or composition described herein in combination with one or more cancer therapies. In some embodiments, the compound or ition is combined with adiation, chemotherapy, and/or radiation therapy. As would be understood by one of skill in the art, radiation” refers to a treatment regime that includes both chemotherapy (such as gemcitabine) and radiation. In some ments, the chemotherapy is gemcitabine.

Yet another aspect provides a method of increasing the sensitivity of pancreatic cancer cells to a cancer therapy selected from gemcitabine or ion therapy by administering a compound or composition described herein in combination with the cancer therapy.

In some embodiments, the cancer therapy is gemcitabine. In other embodiments, the cancer therapy is radiation therapy. In yet another embodiment the cancer therapy is chemoradiation.

Another aspect provides a method of inhibiting orylation of Chkl (Ser 345) in a pancreatic cancer cell comprising administering a compound or composition described herein after treatment with gemcitabine (100 nM) and/or radiation (6 Gy) to a pancreatic cancer cell.

Another aspect provides a method of disrupting damage—induced cell cycle checkpoints by administering a compound or composition described herein in combination with radiation therapy to a cancer cell.

] Another aspect provides a method of inhibiting repair of DNA damage by homologous recombination in a cancer cell by administering a compound or composition described herein in combination with one or more of the following treatments: chemoradiation, chemotherapy, and radiation therapy.

In some embodiments, the chemotherapy is gemcitabine.

Another aspect provides a method of inhibiting repair of DNA damage by homologous recombination in a cancer cell by administering a compound or composition bed herein in combination with gemcitabine and radiation therapy.

Another aspect of the invention provides a method of treating non-small cell lung cancer sing administering to a patient a compound or ition described herein in ation with one or more of the following additional therapeutic agents: tin or Carboplatin, Etoposide, and ionizing radiation. Some embodiments comprise administering to a patient a compound described herein in combination with Cisplatin or latin, Etoposide, and ng radiation. In some embodiments the combination is Cisplatin, Etoposide, and ionizing radiation. In other embodiments the combination is Carboplatin, Etoposide, and ionizing radiation.

] Another embodiment provides a method of promoting cell death in cancer cells comprising administering to a patient a compound described herein, a composition comprising , or said compound.

Yet another embodiment provides a method of preventing cell repair of DNA damage in cancer cells comprising administering to a patient a compound described herein, or a composition comprising said compound. Yet another embodiment provides a method of ting cell repair caused by of DNA damage in cancer cells comprising administering to a patient a nd of the present invention, or composition comprising said compound.

Another embodiment provides a method of sensitizing cells to DNA ng agents comprising administering to a patient a compound described herein, or a composition comprising said compound.

In some embodiments, the method is used on a cancer cell having s in the ATM signaling cascade. In some embodiments, said defect is altered expression or ty of one or more ofthe following: ATM, p53, CHK2, MREl l, RAD50, NBSl, 53BP1, MDCl, H2AX, MCPHl/BRITl, CTIP, or SMCl. In other embodiments, said defect is altered expression or activity of one or more ofthe following: ATM, p53, CHK2, MREl l, RAD50, NBSl, 53BP1, MDCl or H2AX. According to another embodiment, the method is used on a cancer, cancer cell, or cell expressing DNA ng oncogenes.

In another embodiment, the cell is a cancer cell expressing DNA damaging oncogenes. In some embodiments, said cancer cell has altered expression or activity of one or more of the following: K-Ras, N-Ras, H-Ras, Raf, Myc, Mos, E2F, , CDC4, CDK2, Cyclin E, Cyclin A and Rb.

According to another embodiment, the method is used on a cancer, cancer cell, or cell has a defect in a n involved in base excision repair (“base excision repair protein”). There are many s known in the art for determining whether a tumor has a defect in base excision repair. For example, sequencing of either the genomic DNA or mRNA products of each base excision repair gene (e.g., UNG, PARPl, or LIGl) can be med on a sample of the tumor to establish r mutations expected to modulate the function or expression of the gene product are present (Wang et al., Cancer Research 52:4824 (1992)). In addition to the mutational inactivation, tumor cells can modulate a DNA repair gene by hypermethylating its er region, leading to reduced gene expression. This is most commonly assessed using methylation-specific rase chain reaction (PCR) to quantify methylation levels on the promoters of base excision repair genes of interest.

Analysis of base excision repair gene promoter methylation is available commercially (http://www.sabiosciences.com/dna_methylation_product/HTML/MEAH—421A.html).

Finally, the expression levels of base excision repair genes can be ed by directly quantifying levels of the mRNA and protein products of each gene using standard techniques such as quantitative reverse transcriptase-coupled polymerase chain reaction (RT-PCR) and immunhohistochemistry (IHC), respectively (Shinmura et al., Carcinogenesis 25: 2311 (2004); Shinmura et al., Journal of Pathology 225:414 (2011)).

] In some embodiments, the base excision repair protein is UNG, SMUGl, MBD4, TDG, OGGl, MYH, NTH1, MPG, NEIL1, NEIL2, NEIL3 (DNA ylases); APE1, APEX2 (AP endonucleases); LIGl, LIG3 (DNA s I and III); XRCCl (LIG3 accessory); PNK, PNKP (polynucleotide kinase and phosphatase); PARPl, PARP2 (Poly(ADP-Ribose) Polymerases); PolB, PolG (polymerases); FENl (endonuclease) or Aprataxin.

In sorme ments, the base excision repair protein is PARPl, PARP2, or PolB. In other embodiments, the base excision repair protein is PARPl or PARP2.

The methods described above (gene sequence, promoter methylation and mRNA expression) may also be used to characterize the status (e. g., expression or mutation) of other genes or ns of interesting, such DNA—damaging oncogenes expressed bv a tumor or defects in the ATM signaling cascade of a cell.

Yet another embodiment provides use of a nd or composition described herein as a radio-sensitizer or a chemo-sensitizer.

Yet other embodiment provides use of a compound or composition described herein as a single agent (monotherapy) for treating cancer. In some ments, the compounds or compositions described herein are used for treating patients having cancer with a DNA-damage response (DDR) defect. In other ments, said defect is a mutation or loss of ATM, p53, CHK2, MREl l, RAD50, NBSl, 53BP1, MDCl, or H2AX.

Terminology The terms "subject," “host,” or “patient” includes an animal and a human (e. g., male or female, for example, a child, an adolescent, or an adult). Preferably, the "subj ect," “host,” or “patient” is a human.

Compounds of this invention include those bed generally herein, and are further illustrated by the s, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this invention, the al elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Additionally, general principles of organic chemistry are described in ic Chemistry”, Thomas Sorrell, sity Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5th Ed., Ed.: Smith, MB. and March, J., John Wiley & Sons, New York: 2001, the entire ts of which are hereby incorporated by reference.

As described herein, a specified number range of atoms es any integer therein. For example, a group having from 1-4 atoms could have 1, 2, 3, or 4 atoms.

As described herein, compounds of the ion may optionally be substituted with one or more substituents, such as are illustrated generally herein, or as exemplified by particular classes, subclasses, and s of the invention. It will be appreciated that the phrase “optionally substituted” is used interchangeably with the phrase “substituted or unsubstituted.” In general, the term “substituted”, whether preceded by the term “optionally” or not, refers to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Unless ise indicated, an optionally substituted group may have a tuent at each substitutable position of the group, and when more than one on in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds.

Unless ise indicated, a substituent connected by a bond drawn from the center of a ring means that the substituent can be bonded to any position in the ring. In example i below, for instance, JW can be bonded to any position on the pyridyl ring. For bicyclic rings, a bond drawn through both rings indicates that the substituent can be bonded from any position of the bicyclic ring.

In example ii below, for instance, JW can be bonded to the 5-membered ring (on the nitrogen atom, for ce), and to the ered ring. / §_<,Z\n/\\ —|—<JW)o.5 ' (JW)0-5 N H WO 85132 1 11 The term “stable”, as used , refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that is not substantially d when kept at a temperature of 40°C or less, in the absence of moisture or other chemically reactive conditions, for at least a week.

The term “dative bond”, as used herein, is defined as the coordination bond formed upon interaction between molecular species, one of which serves as a donor and the other as an acceptor of the on pair to be shared in the complex .

The term “aliphatic” or “aliphatic , as used herein, means a straight-chain (i.e., unbranched), branched, or cyclic, substituted or tituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation that has a single point of attachment to the rest of the molecule.

Unless otherwise specified, aliphatic groups contain 1-20 aliphatic carbon atoms. In some embodiments, aliphatic groups contain l-lO aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-8 tic carbon atoms. In still other embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. Aliphatic groups may be linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups. Specific examples e, but are not limited to, methyl, ethyl, isopropyl, n-propyl, sec-butyl, vinyl, nyl, ethynyl, and tert-butyl. tic groups may also be , or have a combination of linear or branched and cyclic . Examples of such types of aliphatic groups include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, —CH2— cyclopropyl, CHzCHzCH(CH3)—cyclohexyl.

The term “cycloaliphatic” (or “carbocycle” or “carbocyclyl”) refers to a monocyclic C3—C8 hydrocarbon or bicyclic C8-C12 hydrocarbon that is tely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule wherein any individual ring in said bicyclic ring system has 3-7 members. Examples of cycloaliphatic groups e, but are not limited to, lkyl and cycloalkenyl groups. Specific examples include, but are not limited to, cyclohexyl, cyclopropyl, and cyclobutyl.

The term “heterocycle”, “heterocyclyl”, or “heterocyclic” as used herein means non— aromatic, monocyclic, bicyclic, or tricyclic ring systems in which one or more ring members are an independently selected heteroatom. In some embodiments, the “heterocycle”, “heterocyclyl”, or “heterocyclic” group has three to fourteen ring s in which one or more ring members is a heteroatom independently selected from oxygen, sulfur, nitrogen, or phosphorus, and each ring in the system contains 3 to 7 ring members.

] Examples of heterocycles include, but are not limited to, 3—lH-benzimidazol-2—one, 3-(1- alkyl)-benzimidazolone, 2-tetrahydrofuranyl, 3-tetrahydrofuranyl, ahydrothiophenyl, 3- tetrahydrothiophenyl, holino, 3-morpholino, 4-morpholino, 2-thiomorpholino, 3- thiomorpholino, 4-thiomorpholino, l-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, ltetrahydropiperazinyl , 2-tetrahydropiperazinyl, 3-tetrahydropiperazinyl, l-piperidinyl, 2-piperidinyl, 3-piperidinyl, l-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl, l-piperidinyl, 2-piperidinyl, 3—piperidinyl, 4—piperidinyl, 2—thiazolidinyl, 3—thiazolidinyl, zolidinyl, l—imidazolidinyl, 2— imidazolidinyl, azolidinyl, 5-imidazolidinyl, indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane, benzodithiane, and 1,3-dihydro-imidazolone.

Cyclic groups, (e. g. cycloaliphatic and heterocycles), can be linearly fused, bridged, or yclic.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4- dihydro-2H—pyrrolyl), NH (as in pyrrolidinyl) or NR+ (as in N—substituted pyrrolidinyl)).

The term “unsaturated”, as used , means that a moiety has one or more units of ration. As would be known by one of skill in the art, unsaturated groups can be partially unsaturated or fully unsaturated. Examples of partially rated groups include, but are not limited to, butene, cyclohexene, and tetrahydropyridine. Fully unsaturated groups can be aromatic, anti-aromatic, or non-aromatic. Examples of fully unsaturated groups include, but are not limited to, phenyl, cyclooctatetraene, pyridyl, thienyl, and l-methylpyridin-2(lH)-one.

] The term “alkoxy”, or “thioalkyl”, as used herein, refers to an alkyl group, as previously defined, attached through an oxygen (“alkoxy”) or sulfur (“thioalkyl”) atom.

The terms “haloalkyl”, “haloalkenyl”, “haloaliphatic”, and lkoxy” mean alkyl, alkenyl or alkoxy, as the case may be, substituted with one or more halogen atoms. This term includes perfluorinated alkyl groups, such as —CF3 and —CF2CF3.

] The terms “halogen”, “halo”, and “hal” mean F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in “arylalkyl”, “arylalkoxy”, or “aryloxyalkyl”, refers to monocyclic, bicyclic, and tricyclic ring systems haVing a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term “aryl” may be used interchangeably with the term “aryl ring”.

WO 85132 The term “heteroaryl”, used alone or as part of a larger moiety as in “heteroarylalkyl” or “heteroarylalkoxy”, refers to monocyclic, bicyclic, and lic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and wherein each ring in the system ns 3 to 7 ring members. The term “heteroaryl” may be used interchangeably with the term oaryl ring” or the term “heteroaromatic”. Examples of heteroaryl rings include, but are not limited to, 2-furanyl, 3- furanyl, N—imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, benzimidazolyl, 3-isoxazolyl, 4- isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N—pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2- pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e. g., 3- pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e. g., 5-tetrazolyl), triazolyl (e.g., 2- triazolyl and 5-triazolyl), nyl, nyl, benzofuryl, benzothiophenyl, indolyl (e. g., 2-indolyl), pyrazolyl (e.g., zolyl), azolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, purinyl, pyrazinyl, 1,3,5- triazinyl, quinolinyl (e. g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., l- isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl).

It shall be understood that the term “heteroaryl” includes certain types of heteroaryl rings that exist in equilibrium between two different forms. More specifically, for example, species such hydropyridine and pyridinone (and likewise ypyrimidine and pyrimidinone) are meant to be assed within the definition of “heteroaryl.” |\ |\ /N‘_ NH OH 0 The term “protecting group” and ctive group” as used herein, are interchangeable and refer to an agent used to temporarily block one or more desired functional groups in a compound with multiple reactive sites. In certain embodiments, a protecting group has one or more, or preferably all, of the following characteristics: a) is added selectively to a functional group in good yield to give a protected ate that is b) stable to reactions ing at one or more of the other reactive sites; and c) is selectively removable in good yield by reagents that do not attack the rated, deprotected onal group. As would be understood by one skilled in the art, in some cases, the reagents do not attack other reactive groups in the compound. In other cases, the reagents may also react with other reactive groups in the compound. Examples of protecting groups are detailed in Greene, T.W., Wuts, P. G in “Protective Groups in Organic Synthesis”, Third Edition, John Wiley & Sons, New York: 1999 (and other editions of the book), the entire contents of which are hereby incorporated by reference. The term gen protecting group”, as used herein, refers to an agent used to temporarily block one or more desired nitrogen reactive sites in a multifunctional compound. Preferred nitrogen protecting groups also possess the characteristics exemplified for a protecting group above, and certain exemplary nitrogen protecting groups are also detailed in Chapter 7 in Greene, T.W., Wuts, P. G in “Protective Groups in Organic Synthesis”, Third Edition, John Wiley & Sons, New York: 1999, the entire contents of which are hereby incorporated by reference.

In some embodiments, a methylene unit of an alkyl or tic chain is optionally replaced with another atom or group. Examples of such atoms or groups include, but are not d to, nitrogen, oxygen, , —C(O)—, —C(=N—CN)—, )—, —C(=NOR)—, —SO—, and —SOz—. These atoms or groups can be ed to form larger groups. es of such larger groups e, but are not d to, —OC(O)—, —C(O)CO—, -COz-, —C(O)NR—, —C(=N—CN), —NRCO—, —NRC(O)O— , —, —NRSOz—, —NRC(O)NR—, —OC(O)NR—, and —NRSOzNR-, wherein R is, for e, H or C1_6aliphatic. It should be understood that these groups can be bonded to the methylene units of the aliphatic chain via single, double, or triple bonds. An example of an optional replacement (nitrogen atom in this case) that is bonded to the aliphatic chain via a double bond would be — CHZCH=N—CH3. In some cases, especially on the terminal end, an optional ement can be bonded to the tic group via a triple bond. One example of this would be CHZCHZCHZCEN. It should be understood that in this situation, the al nitrogen is not bonded to another atom.

It should also be understood that, the term “methylene unit” can also refer to branched or substituted methylene units. For example, in an isopropyl moiety [-CH(CH3)2], a nitrogen atom (e. g.

NR) replacing the first recited “methylene unit” would result in dimethylamine [-N(CH3)2]. In instances such as these, one of skill in the art would understand that the nitrogen atom will not have any additional atoms bonded to it, and the “R” from “NR” would be absent in this case.

Unless otherwise ted, the optional replacements form a chemically stable compound.

Optional replacements can occur both within the chain and/or at either end of the chain; i.e. both at the point of attachment and/or also at the terminal end. Two optional replacements can also be adjacent to each other within a chain so long as it results in a chemically stable compound. For example, a C3 aliphatic can be optionally replaced by 2 nitrogen atoms to form —C—NEN. The optional replacements can also completely replace all of the carbon atoms in a chain. For example, a C3 aliphatic can be optionally replaced by —NR-, -C(O)—, and —NR— to form —NRC(O)NR— (a urea).

Unless otherwise indicated, if the replacement occurs at the terminal end, the replacement atom is bound to a hydrogen atom on the terminal end. For example, if a methylene unit of — CH3 were optionally replaced with —O—, the resulting compound could be — OCH2CH3, —CHZOCH3, or —CH2CHZOH. It should be understood that if the terminal atom does not contain any free e electrons, then a hydrogen atom is not required at the terminal end (e.g., —CH2CH2CH=O or —CH2CH2CEN).

Unless otherwise indicated, structures depicted herein are also meant to include all isomeric (e. g., enantiomeric, diastereomeric, ric, conformational, and rotational) forms of the ure. For example, the R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) mational isomers are included in this invention. As would be understood to one skilled in the art, a substituent can freely rotate around any rotatable bonds. For \I N/l example, a substituent drawn as also represents \ Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, geometric, mational, and rotational mixtures of the present compounds are within the scope of the ion.

Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention.

] In the compounds of this invention any atom not specifically ated as a particular isotope is meant to represent any stable isotope of that atom. Unless ise stated, when a position is designated ically as “H” or “hydrogen”, the position is tood to have hydrogen at its natural abundance isotopic composition. Also unless otherwise stated, when a on is designated specifically as “D” or “deuterium”, the position is understood to have deuterium at an abundance that is at least 3340 times greater than the natural abundance of deuterium, which is 0.015% (i.e., at least 50.1% incorporation of deuterium).

“D” and “d” both refer to deuterium.

Additionally, unless otherwise indicated, structures depicted herein are also meant to include compounds that differ only in the ce of one or more isotopically ed atoms. For example, compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13C- or 14C—enriched carbon are within the scope of this invention. Such nds are useful, for example, as analytical tools or probes in biological assays.

As used herein “crystalline” refers to a solid that has a specific arrangement and/or conformation of the molecules in the crystal lattice.

As used herein the term “amorphous” refers to solid forms that consist of disordered arrangements of molecules and do not possess a distinguishable crystal lattice.

As used , the term “solvate” refers to a lline solid adduct containing either stoichiometric or ichiometric amounts of a solvent incorporated within the crystal structure. If the incorporated solvent is water, such adduct is refered to as a “hydrate”.

Abbreviations The following abbreviations are used: DMSO dimethyl sulfoxide DCM dichloromethane ATP adenosine triphosphate TFA trifluoroacetic acid 1HNMR proton nuclear magnetic resonance HPLC high performance liquid chromatography LCMS liquid chromatography-mass spectrometry Rt retention time MTBE Methyl tert—butyl ether XRPD X-Ray Powder Diffraction DSC Differential scanning calorimetry TGA Thermogravimetric is RT room temperature NMP N—methylpyrrolidone Bp boiling point DMF dimethylformamide PTSA p—Toluenesulfonic acid DIPEA N,N—diisopropylethylamine HOBT hydroxybenzotriazole HATU l dimethylamino)methylene]- lH- 1,2,3 -triazolo [4,5-b]pyridinium 3 -oxid hexafluorophosphate TBTU 2-(1 H-benzotriazole- l -yl)- l , 1,3 ,3 -tetramethyluronium tetrafluoroborate T3P phosphonic anhydride COMU l— [( l -(Cyanoethoxyoxoethylideneaminooxy)-dimethylamino- morpholino)]uroniumhexafluorophosphate TCTU [(6-chlorobenzotriazol- l -yl)oxy-(dimethylamino)methylene]-dimethyl-ammonium tetrafluoroborate HBTU O-Benzotriazole-N,N,N’ ,N’ -tetramethyl-uronium-hexafluoro-phosphate ECDI l-Ethyl-3 -(3 hylaminopropyl)carbodiimide LDA Lithium diisopropylamide CD1 1, l'-Carbonyldiimidazole Processes Processes and compounds described herein are useful for producing ATR inhibitors that contain an aminopyrazolopyrimidine core. The l synthetic procedures shown in s herein are useful for generating a wide array of chemical species Which can be used in the manufacture of ceutical compounds.

SCHEMEA NH2 0 O NH2 0 / o anion NC ,AII pyrazole pyrimidine N\ / O’All NCQL condensation 0 formation N// O,AII formation N OAII —’ —>HN _’ \ N H2N CCI3 / 1 R1 2 3 4a-c amide bond NH2 NH 0 2 o N=N NH2 0 . /N R2 / activated ester / N N ,NI amide-bond / O / deprotection / OH formation N / / formation Ni / \ —> —>N —> \ N M R3 <> \ //N g\ //N \J <\ /N R4 5a-c 6a-c l-A Compounds of this invention can be synthesised ing to methods similar to the one depicted in Scheme A.

The anion of commercially available allyl cyanoacetate 1 can react with, e.g., oroacetonitrile to provide intermediate 2. In the anion condensation step, the anion of commercially available allyl cyanoacetate 1 can be generated with a base such as potassium acetate in an appropriate solvent such as an alcohol (e.g., isopropylalcohol). The anion then reacts with trichloroacetonitrile at room temperature.

Step 2 Intermediate 2 then reacts with hydrazine to form the diaminopyrazole 3. In the pyrazole formation step, intermediate 2 is reacted with hydrazine (or its hydrate) in an aprotic solvent, such as 2014/068713 DMF, to provide the diaminopyrazole 3. The reaction occurs under basic conditions (e.g., in the ce of potassium acetate or AcONa) with heating (e. g.,2 110°C) to ensure complete cyclisation.

Step 3 Intermediate 3 can further be condensed with a dielectrophilic coupling partner to form the dine 4a-c. In the pyrimidine formation step, intermediate 3 is d with a 1,3- dielectrophilic species (e. g., a l,3—dialdehyde or a 3-(dialkylamino)—prop—2—enal) in various types of solvents (e. g., DMF or DMSO/water) to furnish the bicyclic cores 4a-c. When one or two of the electrophilic centers is protected/masked (e.g., aldehyde masked as a ketal), introduction of a sulfonic acid (e. g., PTSA) is required to liberate the reactive functional group.

Step 4 Deprotection, e. g, via hydrolysis, of the allyl ester leads to the carboxylic acids Sa-c. In the deprotection step, compound 4a-c is subjected to hydrolytic conditions that are known to those d in the art. For example, treatment of 4a-c with phenylsilane or 4-methylbenzenesulf1nate in the presence of a catalytic amount of palladium (e. g., Pd(PPh3)4) leads to the formation of the corresponding ylic acid 5a-c. Alternatively, compounds 4a-c could be treated with s alkali (e.g., NaOH, LiOH, or KOH) to produce acids 5a-c.

Step 5 In the activated ester formation step, the carboxylic acids Sa-c are reacted with amide coupling agents known to those d in the art. Suitable amide coupling partners include, but are not limited to TBTU, TCTU, HATU, T3P, and COMU. When the coupling agent is chosen appropriately, the reactions can proceed rapidly (~lhr.) at room temperature in the presence of an organic base such as an aliphatic amine (e.g., triethylamine, DIPEA) to provide the activated esters 6a-c. For example, when the amide coupling agents TBTU [J=H] or TCTU[J=Cl] are used, compounds 6a-c are obtained readily by tion of the reaction mixture.

Formation of the activated esters 6a-c prior to the amide bond formation to prepare LA is generally preferred, although a direct conversion of Sa-c into the compounds of formula I-A of this invention is also possible. Alternative activated esters can also be utilised (isolated or formed in situ) and will be known to those skilled in the art (e. g., using CDI, TBTU, TCTU, HATU, T3P, COMU ng agents).

Step_ 6 In the amide bond ion step, activated esters 6a-c can react with a substituted or unsubstituted 3—aminopyridine to provide compounds of formula I-A of this invention. The reaction conditions for the amide coupling are generally in an aprotic solvent (e. g., NMP, pyridine, DMF, etc ...) with heating (e. g., 2 90°C). The 3-aminopyridine may be r funtionalized following amide bond formation.

Alternatively, the two steps described above can be combined: ylic acids 5a-c can be used as starting points for the amide bond formation, the activated esters being generated in situ, using the same amide couplings agents as those described above. Compounds I-A of this invention are isolated in a similar manner to the one described above.

Selle—meB pyrazole NH 'dine O amide bond 0 2 N N 2 o N / / pyrImII R2 0 R ’ R2 formation ion formation ”OJ—’ NC\/[LN\/ —>N/N\// N / I —>\/N\ OH N H H R3 HN H R3 R4 R4 €\ //N R4 ] Alternatively, compounds of the present disclosure can be prepared according to methods similar to the one depicted in Scheme B.

Step 1 The amide 8 can readily be prepared from cially available cetic acid 7. In the amide bond formation step, cyanoacetic acid 7 can react with a substituted 3-aminopyridine to provide compounds 8 of this invention. The reaction conditions for the amide coupling are generally in an aprotic solvent (e.g., DCM, NMP, DMF, etc), in the presence of an organic base, such as an aliphatic amine, (e. g., triethylamine or DIPEA) and an amide ng agent known to those skilled in the art: for example EDCI, TBTU, COMU, T3P, etc....

Step 2 In the pyrazole formation step, the anion of cyanoamide 8 can be generated with a base (such as potassium or sodium acetate) in an appropriate solvent such as an alcohol (e.g., ethanol).

The anion then reacts with trichloroacetonitrile at room temperature. The resulting solid, which can be collected by filtration, is then reacted with hydrazine (or its hydrate) in an aprotic solvent, such as DMF or NMP, to e the diaminopyrazole 9, the latter being r condensed with a trophilic coupling partner to form the pyrimidine portion of the compounds of formula I-A of this invention.

Step 3 In the pyrimidine formation step, intermediate 9 is reacted with a l,3-dielectrophilic species (e. g., a l,3—dialdehyde or a 3—(dialkylamino)—prop—2—enal) in various types of solvents (e.g., iPrOH/water, DMF, or DMSO/water) to furnish the desired products I-A. When one or two of the electrophilic centers is protected/masked (e.g., de masked as a ketal), introduction of a sulfonic acid (e. g., PTSA) is required to liberate the reactive functional group.

PREPARATIONS AND EXAMPLES All commercially available solvents and reagents were used as received. ave reactions were d out using a CEM Discovery microwave. Flash Chromatography, e.g., was carried out on an ISCO© CombiflashR CompanionTM system eluting with a 0 to 100% EtOAc/petroleum ether nt. Other methods known in the art were also utilized to perform Flash Chromotography. Samples were applied pre—absorbed on silica. Where stated, supercritical fluid chromatography (SFC) was performed on a Berger Minigram SFC machine. All 1H NMR spectra were recorded using a Bruker Avance III 500 instrument at 500 MHz. MS s were analyzed on a Waters SQD mass spectrometer with electrospray ionization operating in positive and negative ion mode. Samples were introduced into the mass spectrometer using chromatography. All final products had a purity 295%, unless ied otherwise in the experimental s. HPLC purity was measured on a Waters Acquity UPLC system with a Waters SQD MS instrument equipped with a Waters UPLC BEH C8 1.7 um, 2.1 x 50 mm column and a rd BEH C8 1.7 um, 2.1 x 5 mm guard column.

As used herein, the term n)” refers to the HPLC retention time, in minutes, associated with the compound. Unless otherwise indicated, the HPLC methods utilized to obtain the reported retention times are as described below: HPLC Method Instrument: Waters Acquity UPLC—MS; : Waters UPLC BEH C8 1.7 um, 2.1 x 50 mm with Vanguard BEH C8 1.7 um, 2.1 x 5 mm guard column; Column temperature: 45°C; Mobile Phase A: lOmM ammonium formate in water:acetonitrile 95:5, pH 9; Mobile Phase B: acetonitrile; Detection: 210—400 nm; Gradient: 0-0.40 min: 2% B, 0.40-4.85 min: 2% B to 98% B, 4.85-4.90 min: 98% B to 2% B, 4.90- .00 min: hold at 2% B; Flow rate: 0.6 mL/minute.

Preparation 1: Allyl 3,5-diamino-lH-pyrazolecarboxylate NH2 0 H‘Ni/MO”N ’ / Step I: allyl 3-amin0-4, 4, 4-trichl0r0cyan0but—2-en0ate 2 To a solution of KOAc (589.4 g, 6.006 mol) in isopropanol (3 L) was added allyl cyanoacetate (429.4 g, 403.2 mL, 3.432 mol) and the reaction e was cooled to 5°C.

Trichloroacetonitrile (495.5 g, 3.432 mol) was added in 50 mL portions, maintaining temperature below 15°C. The reaction mixture was then allowed to warm to 20°C and stirred for 3 hr. Water (~4 L) was added to dissolve the inorganic materials and precipitate out the desired product. The mixture was stirred for 20 s and the solid was isolated by filtration under vacuum. This solid was ed, washed with water (2 x 0.5 L) and dried in a vacuum oven overnight at 40°C to afford allyl 3—amino—4,4,4—trichloro—2—cyanobut—2—enoate 2 as an off—white powder (787 g, 85%).

Step 2: Allyl 3, 5-diamin0-IH-pyrazolecarb0xylate 3 To a suspension of allyl 3-amino-4,4,4-trichlorocyano-butenoate 2 (619 g, 2.297 mol) and KOAc (676.3 g, 6.891 mol) in DMF (2.476 L) at 0°C was slowly added hydrazine hydrate (172.5 g, 167.6 mL, 3.446 mol) over 15 min. The reaction mixture was then stirred at ambient temperature for 2 hr., at which stage 1H NMR shows complete consumption of the starting material. on mixture was then heated ght at 110°C before being allowed to cool to ambient and stirred for another 48hr. The e was filtered through a ed glass funnel to remove the precipitated solid and the filtrate was evaporated under reduced re to give a thick liquid. DCM (approx. 2 L) was added, and the mixture filtered again to remove additional solids that have precipitated. The filtrate was purified through a 1 kg silica gel plug (gradient of DCM/MeOH as an eluent), and the solvent was removed to afford an orange solid which was suspended in acetonitrile and heated at about 70°C until all the solid went into solution, at which point the solution was allowed to cool to ambient temperature, then to 2°C. The precipitate that formed was ed by filtration under vacuum, washed with chilled MeCN (~50 mL) and dried to constant mass in a vacuum oven to furnish the title compound as an off-white powder (171.2 g, 41%).

Preparation 2a: 1H-benzo[d] [1,2,3]triazol—1-yl 2-aminofluoropyrazolo[1,5—a]pyrimidine—3- ylate Step I: allyl 2-amin0fluor0-pyrazolo[1,5-a]pyrimidine-S-carboxylate 421 To a suspension of allyl 3,5-diamino-1H-pyrazolecarboxylate 3 (42.72 g, 234.5 mmol) in DMSO (270.8 mL) / Water (270.8 mL), was added p—TsOH hydrate (46.72 g, 245.6 mmol) and 3— (diisopropylamino)—2—fluoro—prop—2—enal (described in Tetrahedron Letters, 33(3), 357—60; 1992) (38.69 g, 223.3 mmol). The reaction mixture was heated to 100°C for 3hr. during which time a solid slowly precipitated out of solution. The orange suspension was allowed to cool down to RT overnight. The solid was filtered, washed with water and dried under vacuum to give allyl 2-amino- 6—fluoro—pyrazolo[1,5—a]pyrimidine—3 —carboxylate 421 as a sand solid (45.05 g, 85% yield).

In an alternative method, allyl 2-aminofluoro-pyrazolo[1,5-a]pyrimidinecarboxylate 421 may be sized via generic Scheme C—1, below.

Scheme C-1 -- NH O electrophlllc 2 RO 0 _, acid fluorinating agent 3 N O —> HO \ H —> HO \ O O H Pyrimidine fi/ F Formation O—\__ 36 38 4a 2014/068713 Reaction 1 Bisacetal protected malonaldehyde (Intermediate 35) may be deprotected under acidic conditions to form ediate 36. In some embodiments, the acidic conditions may be generated by utilizing an acid independently selected from HCl, H2804, MeSOzH, TFA, HBF4, or pTSA in a suitable solvent, e. g., water. ably, the acid used in the reaction is selected from pTSA or MeSOzH. RO is preferably a C1_6aliphatic group. In some embodiments, RO is selected from methyl, ethyl, propyl, isopropyl, butyl or pentyl. In still other embodiments, RO is selected from methyl or ethyl.

Reaction 2 Intermediate 36 may be reacted with an electrophilic fluorinating agent to form intermediate 38. In some embodiments, the electrophilic fluorinating agent is independently selected from l-(Chloromethyl)—4-fluoro-l,4-diazoniabicyclo[2.2.2]octane ditetrafluoroborate (Selectfluor®), Accufluor®, obenzenesulfonamide, substituted l—fluoropyridinium salts, or fluorine gas.

Reaction 3 Intermediate 38 may be reacted with intermediate 3 under suitable condensation ions to form intermediate 43. In some ments, the le condensation conditions may include reacting intermediate 38 with intermediate 3 in the presence of a solvent and heat to furnish the bicyclic core of 4a. The reaction may take place in in various types of solvents, e. g., water, DMSO/water, or DMF.

] In one example, intermediate 4a is formed using the methodology described in Scheme C— Scheme C-2 +/—C| HI}! \ £1 N\ ' O ,N ZBF4 NH _\: NH2 pTSA O F 2 0 o o N, o / \ water V 37 3 [\j / HO H /0 0\ “O \ H Aq.DMSO o H | F 80°C /N \_— 35a 36 38 43 l,l,3,3-tetramethoxypropane 35a (20 g, 121.8 mmol) was dissolved in water (200ml). p— Toluenesulphonic acid monohydrate (23. 17g, 121.8mmol) was added and the mixture stirred at 19- °C for 90 minutes. 1-(Chloromethyl)fluoro-1,4-diazoniabicyclo[2.2.2]octane ditetrafluoroborate 37 (Selectfluor, 1.4 eqv, 60.4g, 170.5 mmol) was added portionwise. The addition was endothermic (201°C to 194°C) however the temperature began to rise slowly once the addition was complete (temp increased to 254°C over 45 minutes). The selectfluor dissolved over 1hr. The mixture was allowed to stir at t temperature for 18hrs. The e was homogeneous after this time.

DMSO (150ml) was added slowly over 5 minutes. The addition was exothermic- the temperature increased from 204°C to 342°C during the addition. The mixture then began to cool. The resulting mixture was stirred for 45 s. Compound 3 (21.4g, 115.7 mmol) was then added portionwise.

The addition was not exothermic. The e was heated to 85°C for 4hrs (LC/Ms profile was identical at 2hr and 4hr time points). The stirred mixture was then allowed to cool to ambient temperature overnight. The ing reaction mixture was a slurry. Water (150ml) was added slowly to the resulting slurry. The temperature increased from 204°C to 215°C. The slurry was stirred for 2 hrs, and then the product was isolated by filtration. The cake was washed with water and dried on the sinter to a beige solid (15.5 g). The product was further dried in a vac oven at 40°C for 20hrs. This gave compound 421 as a beige solid (13.5 g, 50% yield). HPLC purity 97.7 % area; 1H NMR (500 MHz, DMSO—d6) 5 4.83 (2H, d),5.29(1H, d), 5.49(1H, d), .14(1H, m), 6.57 (2H, brs), 8.80 (1H, m), 9.40 (1H, m); 19F NMR (500 MHz, DMSO—d6) 5 .

Step 2: 2-amin0fluor0-pyrazolofl,5-a]pyrimidine-S-carboxylic acid 521 ] To a suspension of allyl 2-aminofluoro-pyrazolo[1,5-a]pyrimidinecarboxylate 4a (45 g, 190.5 mmol) in DCM (1.35 L) was added phenylsilane (41.23 g, 46.96 mL, 381.0 mmol), followed by Pd(PPh3)4 (8.805 g, 7.620 mmol). The reaction was stirred at room temperature for 2hr. 30min. The reaction mixture was filtered and the solid was washed with DCM to give a light yellow solid (43.2g). This solid was triturated further in DCM (225 mL) at RT for 45 min, then filtered and dried overnight under vacuum to provide 2—amino—6—fluoro—pyrazolo[1,5—a]pyrimidine—3—carboxylic acid 521 as a light yellow solid (3 7.77g, 100% yield).

] In an alternative method, sodiummethylbenzenesulfinate (anhydrous, 1.2 eqv, 22.6g, 127mmol) was suspended in dry DMSO (20 vol, 500ml). The stirred mixture was warmed to 30°C under a nitrogen atmosphere. Upon complete dissolution Pd(PPh3)4 (2 mol%, 2.4g, 2.1 mmol) was added. The mixture was stirred for 10 min at 25-30°C after which time a turbid yellow solution was present. Allyl 2-aminofluoro-pyrazolo[1,5-a]pyrimidine-3 -carboxylate 421 (25g, 105.8mmol) was added portionwise, maintaining the ature at 25—30°C. Once addition was complete the cloudy solution was stirred until the reaction was complete by HPLC (2-3 hrs). A heavy precipitate formed after 15 s post addition of the substrate. The mixture became thicker as the on proceeded. The reaction mixture was diluted with water (125 ml) and 2M HCl (66 ml) was added slowly, maintaining the temperature at C. The slurry was stirred for 30 minutes, then filtered.

The filtration was slow (2hrs). The resulting solid was washed with water, then dried on the sinter.

The solid was slurried in DCM (8 vol) for 1hr. The solid was filtered (rapid filtration) and washed with DCM. The solid was re-slurried in chloroform (8 vol) for 1 hr. The acid was filtered and dried on the sinter. It was further dried in a vacuum oven at 50°C for 24 hrs. The product 521 was ed as an off—white solid (18.6g, 85%); 1H NMR (500 MHz, DMSO—d6) 5 12.14 (1H, brs), 9.31 (1H, dd), 8.69 (1H, m), 6.47 (2H, brS); 19F NMR (500 MHz, DMSO—d6) 5 —153.65; MS (ES+) 197.1.

Step 3 .‘ IH-benzo[d][1, 2, 3]triazol—I-yl 2-amin0-6—flu0r0pyrazolo[I, 5-a]pyrimidine-S-carboxylate To a sion of 2—aminofluoro-pyrazolo[1,5-a]pyrimidinecarboxylic acid 521 (20 g, 102.0 mmol) in chloroform (300 mL) was added Et3N (11.35 g, 15.63 mL, 112.2 mmol). The suspension was stirred for ~ 5mins and then (benzotriazol-l-yloxy-dimethylamino-methylene)- dimethyl-ammonium Boron Tetrafluoride was added (32.75 g, 102.0 mmol). The suspension was heated to 60°C for 1hr. before the thick suspension was allowed to cool down to RT. The resulting suspension was filtered, washed with chloroform (200 mL) and dried under vacuum ght to afford the title compound 63 as a light yellow powder (32.5g, 88%).

Preparation 2b: or0benzotriazol—1-yl)aminofluoro-pyrazolo[1,5-a]pyrimidine carboxylate 621* In a 2.5 L three—necked flask equipped with stirrer bar, condenser, nitrogen line and Hanna temperature probe was charged 2-aminofluoro-pyrazolo[1,5-a]pyrimidinecarboxylic acid 521 (60 g, 305.9 mmol), chloroform (900.0 mL) and triethylamine (32.44 g, 44.68 mL, 320.6 mmol). [(6— chlorobenzotriazolyl)oxy-(dimethylamino)methylene]-dimethyl-ammonium (Boron Tetrafluoride Ion (1)) (87.00 g, 244.7 mmol) was added portionwise over 5 mins (internal dropped from 22.7 to WO 85132 21.5°C on complete addition). Mixture heated at 60°C (internal temp) for 2hr., still a cream suspension. e cooled to room temperature then solid collected by filtration, washed well with chloroform (until filtrate runs essentially colourless) and dried by suction to leave product 621* as a cream solid (82.2g, 77% yield). 1H NMR (500 MHz, DMSO-d6) 5 9.55 (dd, 1H), 8.91 (d, 1H), 8.22 (dd, 1H), 8.09 (dd, 1H), 7.57 (dd, 1H) and 6.87 (s, 2H). MS (ES+) 348.1.

In an alternative method, 2-Aminofluoropyrazolo[1,5-a]pyrimidine—3-carboxylic acid 521 (30g, 153 mmol) was slurried in acetonitrile (540ml). Triethylamine (22.5ml, 153mmol) was added, followed by [(6—chlorobenzotriazol-1yl)oxy-(dimethylamino)methylene]-dimethylammonium tetrafluoroborate (TCTU, 54.4g, l). The mixture was stirred at room ature for 2 hrs.

The product was isolated by filtration- the filter cake was washed with acetonitrile l). The product was obtained as a brown solid (49.3 g, 93%); 1H NMR (500 MHz, DMSO—dg) 5 9.55 (dd, 1H), 8.91 (d, 1H), 8.22 (dd, 1H), 8.09 (dd, 1H), 7.57 (dd, 1H) and 6.87 (s, 2H); 19F NMR (500 MHz, 6) 5 —150.1; MS (ES+) 348.1.

Preparation 3: 1H-benzo[d] [1,2,3]triazol—1-yl 2-amin0-6—chloropyrazolo[1,5-a]pyrimidine—3- carboxylate NH2 0 ,N=N Ntil/(0 0,N g\ //N Step I: 2-amin0-6—chlor0-pyrazolo[1,5-a]pyrimidine-S-carboxylate 4b To a suspension of allyl amino-1H-pyrazole—4-carboxylate 3 (1 g, 5.489 mmol) in DMF (5 mL) was added (Z)—2-chlorodimethylamino-propenylidene]-dimethyl-ammonium hexafluorophosphate (1.683 g, 5.489 mmol), followed by triethylamine (722.1 mg, 994.6 uL, 7.136 mmol). The reaction mixture was heated to 60°C for 4hr. during which time a solid slowly precipitated out of solution. The brown suspension was allowed to cool down to RT. The solid was filtered, washed with water and dried under vacuum to give allyl 2-aminochloro-pyrazolo[1,5- a]pyrimidinecarboxylate 4b as a brown solid (1.092 g, 72% yield).

Step 2: 2-amin0-6—chlor0-pyrazolo[1,5-a]pyrimidine-S-carboxylic acid 5b 2014/068713 To a suspension of allyl 2-aminochloro-pyrazolo[1,5-a]pyrimidine-3 -carboxylate 4b (1 g, 3.96 mmol) in DCM (15 mL) was added phenylsilane (856.6 mg, 0.9756 mL, 7.916 mmol), followed by Pd(PPh3)4 (182.9 mg, 0.1583 mmol). The reaction was stirred at room temperature for 7hr. The reaction mixture was filtered and the solid was washed with DCM to give a light yellow solid ). This solid was triturated further in DCM (225 mL) at RT for 45 min, then filtered and dried overnight under vacuum to provide 2-aminochloro-pyrazolo[1,5-a]pyrimidine-3 -carboxylic acid 5b as a yellow solid (791m, 94% yield).

Step 3 .‘ IH-benzo[d][1,2, 3]triazol—I-yl 2-amin0chlor0pyrazolo[1,5-a]pyrimidinecarb0xylate To a solution of 2-aminochloro-pyrazolo[1,5-a]pyrimidinecarboxylic acid 5b (1.51 g, 7.103 mmol) in chloroform (15.1 mL) was added TBTU boron tetrafluoride (2.737 g, 8.524 mmol) and TEA (862.5 mg, 1.188 mL, 8.524 mmol). The reaction mixture was d at 50°C for one hour.

The resulting suspension was filtered, and the solid triturated in ethyl acetate to afford the title compound 6b as a yellow solid (2.05 g, 88%).

Preparation 4: zo[d] [1,2,3]triazol—1-yl 2-amino(cyanomethyl)pyrazolo[1,5- a]pyrimidinecarboxylate NH2 0 IN:N N‘Nj/MO 0,N E\ //N Step I: allyl 2-amin0(cyan0methyl)-pyrazolo[1,5-a]pyrimidine-S-carboxylate 4c To a sion of allyl 3,5-diamino-1H-pyrazole—4-carboxylate 3 (63.49 g, 348.5 mmol) in a mixture of DMSO (340 mL) and water (340 mL), was added ethoxymethyl)-4,4- dimethoxy-butanenitrile (prepared according to Preparation 5, below) (85 g, 418.2 mmol), followed by para—toluene Sulfonic acid e (1) (11.27 g, 59.24 mmol). The reaction mixture was heated to 85°C and stirred overnight. The reaction mixture was cooled with an ice bath. The mixture was diluted with EtOAc (680 mL) and a saturated aqueous solution ofNaHCO3 (1.36 L). The precipitate was filtered and rinsed with water, then with a mixture of water and EtOAc. The brown solid was dried under vacuum to give allyl 2-amino(cyanomethyl)-pyrazolo[1,5-a]pyrimidinecarboxylate 4c as a brown solid (55.94 g, 62% yield).

Step 2: 2-amin0-6—(cyanomethyD-pyrazolofl,5-a]pyrimidine-S-carboxylic acid SC ] To a suspension of allyl 2-amino(cyanomethyl)-pyrazolo[1,5-a]pyrimidine carboxylate 4c (10.2 g, 39.65 mmol) in DCM (350 mL) was added phenylsilane (8.581g, 9.773 mL, 79.3 mmol), followed by Pd(PPh3)4 (1.5 g, 1.298 mmol). The reaction was stirred at room temperature for 2hr. The reaction mixture was filtered and the solid was washed with DCM and dried under vacuum to provide 2-amino(cyanomethyl)-pyrazolo[1,5-a]pyrimidinecarboxylic acid 5c as a yellow solid (8.61g, 100% yield).

Step 3 .‘ IH-benzo[d][1,2, 3]triazol—I-yl 2-amin0(cyanomethybpyrazolofl,5-a]pyrimidine carboxylate 6c To a solution of 2-amino(cyanomethyl)-pyrazolo[1,5-a]pyrimidinecarboxylic acid 5c (5.11 g, 23.53 mmol) in DCM (51 mL) was added TBTU boron tetrafluoride (9.067 g, 28.24 mmol) and TEA (2.858 g, 3.937 mL, 28.24 mmol). The on e was stirred at room temperature for one hour. The resulting sion was filtered, and the solid triturated in hot chloroform to afford the title compound 6c as a beige solid (6.59 g, 84%).

\ N N NH2 I ’ / Pyridine.

|)TFA/DCM (1/3). O .

N=N / N\// N \ / triethylsilane ’ H N N// N \ / N 2 F 90°C,12h / o—N N H F RT, 12h N H N N + —, \ ,N \ /N N s\ N // 56% 96% F F F CI .TFA o OtBu 0 0 0,3” OH 27 28 29 i)TCTU.1.1eq RT, 30mins “Hz 0 N n) / NH2 NH N/ O /N 2 O /N HNflN-CO / 1,4 eq \ / N \ 4M HCI 1.2 sq \_z N H N , F \ / N \ / NMP N / N \ / 25 N N N \ / H RT, 20mins N F H \ ,N N \ /N N RT,30 F 0 mins 100%) F F —> O -TFA I» O .HCI 647 0 ° OH OH LN 1-1 Step I: tert—butyl I-[3-[(2-amin0-6—fluor0-pyrazolofl,5-a]pyrimidine-S-carbonybamin0]flu0r0- 4-pyridyl]pmeridine-él-carboxylate 28 A mixture of (6-chlorobenzotriazolyl) 2-aminofluoro-pyrazolo[1,5-a]pyrimidine carboxylate 621* (44.02 g, 126.6 mmol) and tert—butyl 1-(3-aminofluoropyridyl)piperidine ylate 27 (prepared according to Preparation 7b) (34 g, 115.1 mmol) in pyridine (510.0 mL) was heated at 95°C internally overnight (18hr.). Mixture was cooled to room temperature (product precipitated) then added ethanol (340.0 mL) and stirred at room temperature for 10 mins. Collected yellow solid by filtration, washed well with ethanol, dried by suction, then on high vac line for 1hr. to leave t 28 as a yellow solid, (32.5g 56% yield). 1H NMR (500 MHz, DMSO—d6) 5 10.45 (s, 1H), 9.58 (s, 1H), 9.51 (dd, 1H), 8.72 (dd, 1H), 8.25 (d, 1H), 6.81 (s, 2H), 3.15 — 2.93 (m, 4H), 2.55 — 2.47 (masked , 1H), 2.02 — 1.91 (m, 4H), 1.47 (s, 9H). MS (ES+) 474.2.

Step 2: I-[3-[(2-amin0flu0r0-pyrazolofl,5-a]pyrimidine-S-carbonyl)amin0]flu0r0 pyridyUpiperidinecarb0xylic acid trifluorocetate 29 To a suspension of tert—butyl 1—[3—[(2—aminofluoro-pyrazolo[1,5—a]pyrimidine—3— carbonyl)amino]fluoropyridyl]piperidinecarboxylate 28 (69.7 g, 147.2 mmol) in DCM (348.5 mL) and triethylsilane (18.83 g, 25.87 mL, 161.9 mmol) was added TFA (151.1 g, 102.1 mL, 1.325 mol) (mixture sets solid on l addition of TFA then goes into solution after complete addition). ing orange solution was stirred at room temperature overnight. onal TFA (16.78 g, 11.34 mL, 147.2 mmol) was added and the mixture stirred at room temperature for 2hr.

Mixture then heated at 40°C for 20 mins to force reaction to completion. Mixture was trated in vacuo, chloroform (300 mL) was added and mixture again concentrated in vacuo to leave an orange solid suspension. Mixture triturated in DCM (approx. 200 mL), stirred for 20 mins then solid collected by filtration, washed with l DCM and dried by suction to leave a yellow solid.

Filtrate was concentrated in vacuo, residue re-slurried in DCM (approx. 50 mL), stirred for 20 mins then solid collected by filtration, washed with minimal DCM and dried by n to leave a yellow solid which was combined with first crop of solid. Solid dried under vacuum overnight to leave desired product 29 as a yellow solid (82.8g, 96%). 1H NMR (500 MHz, DMSO—d6) 5 10.44 (s, 1H), 9.59 (s, 1H), 9.50 (dd, 1H), 8.84 (dd, 1H), 8.33 (d, 1H), 3.13 — 3.10 (m, 4H), 2.57 — 2.47 (masked , 1H) and 2.08 — 1.93 (m, 4H). MS (ES+) 418.1.

Step 3: I—[3-[(2-amin0-6—flu0r0-pyrazolofl,5-a]pyrimidinecarb0nyl)amin0]flu0r0 pyridyUpzperidinecarb0xylic acid hydrochloride 30 To a solution of 1-[3-[(2-aminofluoro-pyrazolo[1,5-a]pyrimidine-3 -carbonyl)amino] fluoropyridyl]piperidinecarboxylic acid (Trifluoroacetic Acid) 29 (73 g, 124.7 mmol) in NMP (662.7 mL) was added hydrogen chloride (4M in 1,4—dioxane) (37.40 mL of 4 M, 149.6 mmol). After a few seconds a yellow precipitate formed. Mixture stirred at room temperature for 20 mins, then solid collected by filtration, washed with minimal NMP then MTBE, and dried by suction to leave pure t 30 as a light yellow solid, (59.7g, quantitative yield). MS (ES+) 418.1.

Step 4: 2-amin0-6—flu0r0-N-[5-flu0r0-4—[4-[4—(oxetan-S-priperazine-I—carbonyU—I—piperidyU-S- pyridyUpyrazolofl,5-a]pyrimidine—S-carboxamide (Compound I-1) To a yellow suspension of 1—[3—[(2—amino—6—fluoro—pyrazolo[1,5—a]pyrimidine—3— carbonyl)amino]fluoropyridyl]piperidinecarboxylic acid (Hydrochloric Acid) 30 (59.7 g, 131.5 mmol) in NMP (477.6 mL) was added DIPEA (50.99 g, 68.72 mL, 394.5 mmol) then [(6— chlorobenzotriazolyl)oxy-(dimethylamino)methylene]-dimethyl-ammonium (Boron Tetrafluoride Ion (1)) (51.44 g, 144.7 mmol). A yellow sion forms after a few minutes. The mixture was sirred for 30 mins at room temperature then 1-(oxetanyl)piperazine 25 red according to Preparation 8, below) (26.18 g, 184.1 mmol) was added. The cream/tan suspension turns to an orange solution (exotherms from 23.9 to 294°C). The flask was placed on ice/water bath until internal temperature was at 24°C, then ice bath was removed and al temperature steady at 24°C thereafter.

WO 85132 The solution was stirred for 30 mins at room temperature then cooled on an ice/salt/water bath to 10°C before the slow addition of water (1.015 L) in 100 mL portions. Prior to adding the next 100mL of water, waited for exotherm to between 17°C and 20°C (internal) then allow to cool to between 10 and 15°C. Repeated until all water added. Once exotherm had ceased, lt/water bath removed and mixture stirred at ambient temperature for 20 mins (thick /cream suspension . Solid collected by filtration through a sinter funnel, washed well with water then dried by suction for 10 mins. Vacuum d and solid slurried in water on sinter funnel, then vacuum reapplied and solid dried by suction overnight then dried in vacuum oven for 24 h at 40°C <10 mBar.

Solid (54.5 g) suspended in l (545 mL, 10 vol eq.) and heated under reflux for 2hr. then cooled to room temperature over 2h. Solid collected by filtration, washed with minimum ethanol and dried by suction for 1h to leave product as a pale yellow solid. Solid placed in vacuum oven at 23.5°C and <10mBar overnight to leave the ethanol solvate solid form of LI as a pale yellow solid, (51g, 64% yield). 1H NMR (500 MHz, 6) 5 10.64 (s, 1H), 9.67 (s, 1H), 9.48 (dd, 1H), 9.26 (dd, 1H), 8.26 (d, 1H), 6.79 (s, 2H), 4.55 (t, 2H), 4.47 (t, 2H), 4.34 (t, 0.7H), 3.61 (dt, 4H), 3.48 — 3.41 (m, 25H), 3.22 — 3.17 (m, 2H), 3.05 — 3.03 (m, 2H), 3.99 — 2.93 (m, 1H), 2.28 (dt, 4H), 2.17 — 2.10 (m, 2H), 1.74 (d, 2H), 1.07 (t, 2H). MS (ES+) 542.3.

S} E 1 NH2 0 / N d N NH / NH NH2 0 N=N 2 O 2 O / O O/i< N/ NQF HCI I O I / \ N/ \ ,N 27 \ / H dioxane/ N F N’ / N F N O N N water \ H 25 \ / H N / —» —>N N —> N N N ° \ / >—’ />\ N TCTU Pyndme,9oc \ IN CI // -HC' DlPEA \ /N F THF F O O 4\ 0 OH 0 N/fi 6a* \C\O 28 30 [.1 Step I: tert—butyl I-(3-(Z-amino-6—fluor0pyrazolofl,5-a]pyrimidine-S-carboxamido) fluoropyridin-4—yl)pzperidinecarb0xylate 28 6-chloro-1H—benzo[d][1,2,3]triazolyl 2-aminofluoropyrazolo[1,5-a]pyrimidine-3 - carboxylate 621* (45g, 129.4mmol) and tert—butyl 1-(3-aminofluoropyridinyl)piperidine carboxylate 27 (prepared according to Preparation 7b, described below) (40.1g, 135.9mmol) were slurried in pyridine (675ml). The mixture was heated at 95°C under nitrogen until the reaction was complete (determined by HPLC analysis). The mixture was cooled and ethanol (450ml) was added dropwise. The e was filtered and the filter cake washed with ethanol l). The damp cake was dried to give the product 28 as a yellow crystalline solid , 78%); 1H NMR (500 MHz, DMSO—d6) 5 10.45 (s, 1H), 9.58 (s, 1H), 9.51 (dd, 1H), 8.72 (dd, 1H), 8.25 (d, 1H), 6.81 (s, 2H), 3.15 — 2.93 (m, 4H), 2.55 — 2.47 (masked signal, 1H), 2.02 — 1.91 (m, 4H), 1.47 (s, 9H); 19F NMR (500 MHz, DMSO—d6) 5 — 153.5, —136.3; MS (ES+) 474.2.

In an alternative embodiment, intermediate 28 may be purified prior to performing step 2 by using a procedure similar to the following: Step Ia: Purification -butyl (2-amino-6—fluoro-pyrazolo[1,5-a]pyrimidine carbonyl)amino]fluoropyridyl]pzperidinecarboxylate 28 tert—Butyl 1—[3-[(2-aminofiuoro-pyrazolo[1,5-a]pyrimidinecarbonyl)amino] fiuoropyridyl]piperidinecarboxylate 28 (530g; 1.12moles) was added to a mixture ofNMP (5.3L) and 1,2-diaminopropane (249g; 3.3 6moles) and the resulting thin suspension was stirred at —25°C for 15 hours. Ethanol (10.4L) was added to the suspension and the suspension was stirred for 4 hours at 20-25°C. The crystalline golden solid was collected by filtration, washed with ethanol (2 x 2.6L), dried by suction then dried in a vacuum oven for 24 hours at 35—40°C to give 28 as a crystalline golden solid (479g; 90%). 1H NMR (500 MHz, DMSO—d6) 5 10.45 (s, 1H), 9.57 (s, 1H), 9.49 (dd, 1H), 8.71 (d, 1H), 8.24 (d, 1H), 6.79 (s, 2H), 3.44 — 3.33 (m, 1H), 3.34 — 3.20 (m, 4H), 3.07 (dt, 4H), 2.01 — 1.89 (m, 4H), 1.46 (s, 9H). 191: NMR (500 MHz, DMSO—d6) 5 -136.3, —153.4.

Step 2: I-(3-(2-amino-6—fluoropyrazolo[1,5-a]pyrimidinecarboxamido)fluoropyridin yl)pzperidinecarboxylic acid hydrochloride 30 Tert—butyl 1—(3-(2-aminofiuoropyrazolo[1,5-a]pyrimidinecarboxamido) yridinyl)piperidinecarboxylate 28 (36g, 76mmol) was suspended in a solution of HCl in 1,4—dioxane (4M, 670ml). Water (36ml) was added dropwise to the rapidly stirred slurry. The mixture was stirred under nitrogen until the on was complete mined by HPLC is).

The mixture was diluted with 1,4-dioxane (180ml) and filtered. The filter cake was washed with TBME (2x72ml). The damp cake was dried to give a pale brown solid (hydrochloride salt, 32.7g, 95%); 1H NMR (500 MHz, DMSO—d6) 5 10.34 (s, 1H), 9.53—9.49 (m, 2H), 8.82 (m, 1H), 8.50 (m, 1H), 3.13 — 3.22 (m, 4H), 2.57 — 2.47 (masked signal, 1H) and 2.08 — 1.93 (m, 4H); 19F NMR (500 MHz, DMSO—d6) 5 — 152.9, —133.8; MS (ES+) 418.1.

Step 3 .‘ 0-6—flu0r0-N-[5-flu0r0-4—[4-[4—(oxetan-S-priperazine-I—carbonyU—I—piperidyU-S- pyridyljpyrazolofl,5-a]pyrimidine-S-carboxamide (Compound Iamorphous) To a solution of 1-(oxetanyl)piperazine 25 (525mg, 3.69mmol) in THF (12ml) was added DIPEA (1.72ml, 9.91mmol), followed by 1-(3-(2-aminofluoropyrazolo[1,5-a]pyrimidine carboxamido)fluoropyridinyl)piperidinecarboxylic acid (hydrochloride salt, 1.5g, 3.3mmol). [(6-chlorobenzotriazolyl)oxy-(dimethylamino)methylene]-dimethyl-ammonium tetrafluoroborate (TCTU, 1.29g, 3.64mmol) was added and the mixture d under nitrogen until reaction completion (determined by HPLC analysis). The mixture was cooled and water (24ml) was added dropwise. The mixture was allowed to warm to ambient and stirred for 3 hrs, then filtered. The filter cake was washed with (3x3 ml). The damp cake was dried under vacuum (with a nitrogen bleed) at 40°C. An amorphous form of compound I-1 was obtained. (1.54g, 86%); 1H NMR (500 MHz, DMSO—d6) 5 10.64 (s, 1H), 9.67 (s, 1H), 9.48 (dd, 1H), 9.26 (dd, 1H), 8.26 (d, 1H), 6.79 (s, 2H), 4.55 (t, 2H), 4.47 (t, 2H), 4.34 (t, 0.7H), 3.61 (dt, 4H), 3.48 — 3.41 (m, 2.5H), 3.22 — 3.17 (m, 2H), 3.05 — 3.03 (m, 2H), 3.99 — 2.93 (m, 1H), 2.28 (dt, 4H), 2.17 — 2.10 (m, 2H), 1.74 (d, 2H), 1.07 (t, 2H); 19F NMR (500 MHz, DMSO—d6) 5 — 152.8, ; MS (ES+) 542.3.

Compound I-1°amorphous may be ed using an alternative method from Example 2, Step 3, above.

In another example, nd Iamorphous was prepared by adding N,N— Diisopropylethylamine (461uL; 342mg; 2.64mmol) to a suspension of 1-[3-[(2-amino—6—fluoro— pyrazolo[1,5 -a]pyrimidine-3 -carbonyl)amino] -5 -fluoropyridinyl]piperidinecarboxylic acid hydrochloride 30 (1.00g; 2.20mmol; LR) in THF (20mL). 1,1'-Carbonyldiimidazole (CD1) (430mg; 2.65mmol) was added and the mixture was heated at 40-50°C. Additional charges of 1,1'— Carbonyldiimidazole (CD1) (213mg total; 1.31mmol) were made and the e heated until reaction completion (determined by HPLC analysis). 1-(oxetan-3 -yl)piperazine 25 (375mg; 2.64mmol) was added and the mixture was heated at 55-60°C until reaction completion mined by HPLC analysis). The reaction was cooled to C. Water (40mL) and 2M NaOH(aq) (551uL) were added and the suspension was d for 5—10 minutes. The solids were collected by filtration, washed with water (2 x 5mL), dried by suction then dried in a vacuum oven at 45—50°C for 16 hours to give 1—1 as a yellow solid (869mg; 73%).

Pre aration 5: Alternative a roach to s nthesis of ut l 1- 3- 2-amino fluoro razolo 1 5—a rimidine—3-carboxamido fluoro ridin l i eridine—4- carboxylate (Compound 281 / m N OH SOCIZ, Et3N / 27 N CI N ‘N ’ —.' ‘N / f” \ x” CHZCIZ, 40 C \ /N \ /N Pyrldlne, 90°C F F 53 34 28 Step I: 2-amin0-6—fluor0-pyrazolofl,5-a]pyrimidine-S-carbonyl chloride 34 To a suspension of 2—amino-6—fluoro—pyrazolo[1,5—a]pyrimidine—3—carboxylic acid 521 (500mg, 2.55mmol) in dichloromethane (7.5mL) was added triethylamine (409uL, 297mg, 2.93mmol). Thionyl chloride (205uL, 334mg, 2.80mmol) was added and the mixture heated at 35- 40°C for 2 hours. The mixture was cooled to ambient temperature and d at ambient temperature until reaction completion (monitored by HPLC). The solid was collected by ion, washed with dichloromethane (2 x 1mL) and dried by suction to give the product 34 as a beige solid (465mg, 85%); 1H NMR (500 MHz, DMSO—d6) 5 9.30 (dd, J= 4.9, 2.7 Hz, 1H), 8.68 (d, J= 2.7 Hz, 1H); 19F NMR (500 MHz, DMSO—d6) 5 —154. 1.

Step 2: tert—bulyl I-(3-(2-amin0-6—fluor0pyrazolofl,5-a]pyrimidinecarb0xamid0)flu0r0pyridin- 4-yl)pzperidine—4-carb0xylate 28 2-aminofluoro-pyrazolo[1,5-a]pyrimidinecarbonyl chloride 34 (100mg, 0.466mmol) and tert—butyl minofluoropyridinyl)piperidinecarboxylate 27 (138mg, 0.466mmol) were slurried in pyridine (1.5mL). The mixture was heated to 90-100°C for 16 hours. The mixture was cooled and ethanol (3mL) was added. The e was d for 1—2 hours, filtered and the filter cake washed with ethanol (0.5mL). The solids were dried by suction to give the product 28 , 73%). 1H NMR (400 MHz, DMSO—d6) 5 10.45 (s, 1H), 9.57 (s, 1H), 9.50 (dd, J= 4.8, 2.5 Hz, 1H), 8.71 (d, J=2.5 Hz, 1H), 8.24 (d, J= 2.5 Hz, 1H), 6.80 (s, 2H), 3.07 (dd, J= 6.5, 3.3 Hz, 4H), 2.11 — 1.80 (m, 4H), 1.46 (s, 9H); 19F NMR (500 MHz, 6) 5 —136.8, —153.9; MS (ES+) 474.2. 1C0mpound I-lz HZN F H N E J ” N 5N OHF/NI o 6 NH 2/ NH2 O / NH2 0 N \ MsOH I MeCN N O,N N F / N H N —> N‘fi/k N \>_//N HPyndine 90°C FSN:N 339:: THF $_//N F .MSOHO O N/fi 6a* \C\O 28 33 1.1 Step I: tert—bulyl I-(3-(2-amin0-6—fluor0pyrazolofl,5-a]pyrimidinecarb0xamid0)flu0r0pyridin- 4-yl)pzperidine—4-carb0xylate 28 ro-1H—benzo[d][1,2,3]triazolyl 2-aminofluoropyrazolo[1,5-a]pyrimidine-3 - carboxylate 621* (45g, 129.4mmol) and tert—butyl 1-(3-aminofluoropyridinyl)piperidine carboxylate 27 (prepared accoding to Preparation 7b, described below) (40.1g, 135.9mmol) were slurried in pyridine (675ml). The mixture was heated at 95°C under nitrogen until the reaction was complete (determined by HPLC analysis). The mixture was cooled and ethanol (450ml) was added dropwise. The mixture was filtered and the filter cake washed with ethanol (2x70ml). The damp cake was dried to give the product 28 as a yellow crystalline solid (47.7g, 78%); 1H NMR (500 MHz, DMSO—dg) 5 10.45 (s, 1H), 9.58 (s, 1H), 9.51 (dd, 1H), 8.72 (dd, 1H), 8.25 (d, 1H), 6.81 (s, 2H), 3.15 - 2.93 (m, 4H), 2.55 — 2.47 (masked signal, 1H), 2.02 — 1.91 (m, 4H), 1.47 (s, 9H); 19F NMR (500 MHz, DMSO—d6) 5 — 153.5, —136.3; MS (ES+) 474.2.

Step 2: I-[3-[(2-amin0flu0r0-pyrazolofl,5-a]pyrimidine-S-carbonyl)amin0]flu0r0 pyridyl]piperidinecarb0xylic acid mesylate 33 Methanesulphonic acid (274uL; 406mg; 4.22mmol) was added to a sion of tert— butyl 1-[3-[(2-aminofluoro-pyrazolo[1,5-a]pyrimidine-3 -carbonyl)amino]fluoro pyridyl]piperidinecarboxylate 28 (1.00g; 2.11mmol) in acetonitrile (15mL) and the e was heated to 75—80°C for 16 hours. The solids were collected by ion, washed with acetonitrile (2 x 2mL) and dried under vacuum to give 1-[3-[(2-aminofluoro-pyrazolo[1,5-a]pyrimidine carbonyl)amino]fluoropyridyl]piperidine—4—carboxylic acid mesylate 33 (0.94g; 87%). 1H NMR (500 MHz, DMSO—d6) 5 10.43 (s, 1H), 9.58 (s, 1H), 9.49 (dd, 1H), 8.83 (d, 1H), 8.32 (d, 1H), 6.85 (bs, 2H), 3.11 (dt, 4H), 2.31 (s, 3H), 1.99 (m, 4H); 19F NMR (500 MHz, 6) 5 —135.5, — 153.1; MS (ES+) 418.1.

Step 3 .‘ 2-amin0-6—fla0r0-N-[5-fla0r0[4-[4-(oxetan-S-priperazine-I-carbonyU—I-piperidyl] pyridyljpyrazolofl,5-a]pyrimidine—S-carboxamide (Compound I-1 amorphous) MN—Diisopropylethylamine (51uL; 38mg; 0.29mmol) was added to a suspension of 1—[3— [(2-aminofluoro-pyrazolo[1,5-a]pyrimidinecarbonyl)amino]fluoropyridyl]piperidine ylic acid mesylate (50mg; 0.097mmol) and 1-(oxetanyl)piperazine (15mg; 0.11mmol) in THF (1.00mL). [(6-chlorobenzotriazolyl)oxy-(dimethylamino)methylene]-dimethyl-ammonium tetrafluoroborate (TCTU, 36.3mg; ol) was added and the mixture stirred under nitrogen until reaction tion (determined by HPLC analysis). Water (2mL) was added to the suspension and stirred for 5 hours. The solids were collected by filtration, washed with water (2 x 200uL), dried by suction then dried in a vacuum oven for 24 hours at 45—50°C to give I-1 as a pale yellow solid (3 1mg; 59%).

Pre n 6: Pre aration of butanenitrile intermediates OMe OMe OMe OMe functional MeO OMe MeO OMe M90 O'V'e M60 OMe group alkylation ’ + transformation OMe OMe R8 OMe R88 OMe OH ON ON CN 11 12a (R8=Me) 13a (R8=Me) 12b (R8=Et) 13b ) Step I: 3-(dimeth0xymethyD-4,4-dimeth0xybatanenitrile 11 2—(dimethoxymethyl)—3,3—dimethoxy—propan—1—ol 10 al 0fthe American Chemical y (1973), 95(26), 8741) (92 g, 473.7 mmol) was dissolved in dry THF (920 mL) and the mixture was cooled down with an ice bath. Triethylamine (143.8g, 198.1 mL, 1.421 mol) was added at once, followed by dropwise addition of methane sulfonyl chloride (59.69g, 40.33mL, 521.1 mmol), over 1hr. and keeping the internal temperature below 5°C. The reaction mixture was stirred for 1 hr. and then allowed to warm to room temperature. The mixture was diluted with ethyl acetate (920mL) and water (920mL). The layers were separated and the organic layer was isolated, washed with a saturated solution ofNaHCO3, then brine. The organics were dried over MgSO4, ed and ated to give [2-(dimethoxymethyl)-3,3-dimethoxypropyl]methanesulfonate as an orange oil (125.31g, 97%) which was used directly without further purification.

Tetraethylammonium cyanide (142.3g, 910.8mmol) was added portionwise over 10 minutes to a solution of methoxymethyl)-3,3-dimethoxypropyl]methanesulfonate (124g, 455.4mmol) in MeCN (1.24L). The reaction mixture was stirred at room temperature for 72hr., then ned between ethyl acetate (1 .24L) and water (1 .24L). The layers were separated and the organic layer was isolated, washed with brine. The organics were dried over MgSO4, ed and evaporated to give 3-(dimethoxymethyl)-4,4-dimethoxybutanenitrile 11 as a dark brown oil (86. lg).

Step 2: 3-(dimeth0xymethyD-4,4-dimeth0xymethylbutanenitrile 12a and 3-(dimeth0xymethyD-4,4- dimethoxy-Z,2-dimethylbutanenitrile 13a To a solution of 3-(dimethoxymethyl)-4,4-dimethoxy-butanenitrile 11 (250 mg, 1.205 mmol) in THF (3 mL) at —75°C was added a solution of iodomethane (513.1 mg, 225.0 uL, 3.615 mmol) in THF (1 ml). A THF solution of (bis(trimethylsilyl)amino)sodium (1.808 mL of 2M, 3.615 mmol) was then added, keeping the temperature below -60°C. After addition, the reaction mixture was stirred at —75°C for 2hrs and then slowly quenched with s saturated NH4C1 solution (5ml).

The mixture d with water and ether and layers separated. The organic layer was washed with brine, dried 4) and concentrated in vacuo to afford a yellow oil which was purified by chromatography on silica gel, eluting with a petroleum ether:EtOAc gradient of 100:0 to 80:20.

Solvents were concentrated in vacuo to afford a clear oil (194mg). NMR proved this oil to be a mixture of 80% mono methyl compound 12a with and 20% bis methyl compound 13a. This mixture was used directly in subsequent steps.

Step 3: 3-(dimethoxymethyl)ethyl—4,4-dimeth0xybutanenitrile 12b and 3-(dimeth0xymethyD diethyl—4, 4-dimeth0xybutanenitrile 13b When ethyl iodide was used instead of methyl iodide in a r procedure to Preparation 6, step 2, above, a e of monosubsituted compound 12b and disubstituted compound 13b was isolated and used directly in subsequent steps.

Pre aration 7a: S nthesis of tert—but 11- 3-amino-5—fluoro rid l i eridinecarbox late i) PhZC=NH o OtBu N\ Pd2(dba)3, xantphos N\ NaZC03 ) | C32003, dioxane I N\ / / cyclohexanol 8.5 vol eq Br F 95°C 12h H2N F | 120°C, 24h / N ii) 2M HCI/THF, rt N Br F 0' 49% 83% .HCI o OtBu o OtBu 26 27 Step I: tert—butyl I-(3-br0m0fluor0pyridyl)piperidinecarb0xylate 26 A 3L flange flask equipped with a thermometer, condensor, en line and overhead stirrer was heated at 40°C (external) then charged with cyclohexanol (750 mL), disodium carbonate (129.8 g, 1.225 mol), 3-bromo-4—chloro—5—fluoro—pyridine (Hydrochloric Acid 18) (137.5 g, 556.8 mmol) and utyl piperidinecarboxylate (123.8 g, 668.2 mmol) rinsed in with cyclohexanol (350 mL). Mixture was heated to 120°C internal temperature overnight (18hr.). Reaction mixture was removed from hotplate and allowed to cool to room temperature. Water (687.5 mL) and EtOAc (687.5 mL) were added, stirred for 10 mins then transferred to separating funnel. Additional EtOAc (1.238 L) was added, mixed and aqueous phase was removed. Organic phase was further washed with water (687 mL), aqueous phase removed, c layer collected. Aqueous phases were combined and back extracted with EtOAc (687.5 mL), aqueous layer removed and organic phase combined with other organics. Organics concentrated in vacuo (water bath temp = 60°C, vacuum down to 2 mBar) leaving a viscous brown oil.

Oil was dissolved in 25% EtOAc/petrol then passed through a short silica pad, g with % EtOAc/petrol until no more product came off. Filtrate was concentrated in vacuo to leave a brown oil, . Product re—purified by ISCO companion (1.5Kg , loaded in DCM, eluting 0 to 20% EtOAc/petrol), product fractions combined and trated in vacuo to leave d product 26 as a pale yellow to cream solid, (98g, 49% yield). 1H NMR (500 MHz, DMSO—d6) 5 8.47 (s, 1H), 8.41 (d, 1H), 3.39 — 3.36 (m, 2H), 3.12 (tt, 2H), 2.49 —2.43 (m, 1H), 1.91 — 1.87 (m, 2H), 1.71 — 1.64 (m, 2H) and 1.43 (s, 9H). MS (ES+) 361.0.

Step 2: tert—butyl I-(S-aminofluor0pyridyl)piperidinecarb0xylate 27 To a solution of tert—butyl l—(3—bromofluoropyridyl)piperidinecarboxylate 26 (98 g, 272.8 mmol), diphenylmethanimine (59.34 g, 54.94 mL, 327.4 mmol) and CszCO3 (177.8 g, 545.6 mmol) in 1,4-dioxane (1.274 L) was added Xantphos (15.78 g, 27.28 mmol) and Pd2(dba)3 (12.49 g, 13.64 mmol). The mixture was stirred under nitrogen at 95°C overnight. The e was cooled to room temperature then partitioned between EtOAc (1000 mL, 10 vol eq.) and water (490 mL, 5 vol eq.), mixed and c layer separated. Organics washed further with water (1 x 250 mL), brine (250 mL), dried (MgSO4), filtered and concentrated in vacuo to leave crude product as a dark red viscous oil, .

The obtained product oil g) was dissolved in THF (882.0 mL) and HCl (545.5 mL of 2 M, 1.091 mol) was added. The resulting mixture was stirred at room ature for 20 mins.

THF was removed in vacuo then additional (HCl (2M) (588.0 mL) was added. The aqueous was washed twice with EtOAc (294.0 mL). A large amount of a yellow itate formed during extraction in both organic and aqueous phase, the solid from both the organic and aqueous phase was collected by filtration and dried by suction. The mixed organic and aqueous filtrate was added to separating funnel, extracted with 2M HCl (2 x 200 mL). All aqueous phases plus solid collected on sinter (product) were combined to give a suspension. The pH was adjusted to 6 using 2M NaOH and extracted with DCM (3 x 600 mL). The organics were combined, dried (MgSO4), filtered and concentrated in vacuo to leave a pale orange waxy solid, 112.2g. This solid was slurried in MeCN (200 mL), stirred for 10 mins then solid collected by filtration, washed with minimal MeCN and dried by suction to leave product 27 as a white solid (66.8 g, 83% yield). 1H NMR (500 MHz, DMSO—d6) 5 7.82 (d, 1H), 7.63 (d, 1H), 5.22 (s, 2H), 3.11 — 3.00 (m, 2H), 2.91 (tt, 2H), 2.36 (tt, 1H), 1.88 — 1.83 (m, 2H), 1.79 — 1.71 (m, 2H), 1.43 (s, 9H). MS (ES+) 297.1.

Scheme 7b: ative a roach to s nthesize tert-but l 1- 3-aminofluoro-4— rid l i eridinecarbox late DIPA thCNH (CCI3)2 Br dba)s N N KF DMSO \ xanphos F|/ I \ I Me4NCI / / 4M HCI F Br F Br 100c Br DIPEA 2MeTHF room temp 31 18 32 26 270k Step I: 3-br0m0chlorofluor0pyridine hydrochloride 18 A solution of diisopropylamine (101.2 g, 140.2 mL, 1.000 mol) in tetrahydrofuran (1.148 L) was cooled to between —25°C and —20°C. ithium (2.5M in hexanes) (400 mL of 2.5 M, 1.000 mol) was added at such a rate as to maintain the reaction temperature below -20°C (addition 20 minutes). The mixture was then allowed to warm to 4°C over 1 hour, then re—cooled to —78°C. 3— bromo—5—fiuoro—pyridine (153.0 g, 869.6 mmol) in tetrahydrofuran (3 82.5 mL) was added over 40 minutes. The mixture was stirred for 90 minutes, then a solution of 1,1,1,2,2,2-hexachloroethane (205.9 g, 869.6 mmol) in tetrahydrofuran (350.0 mL) was added dropwise over 40 minutes. Once the on was complete the mixture was allowed to warm to ambient overnight. The mixture was cooled to 0°C then transferred into cold water (2 L), stirred for 20 mins then MTBE (2.5 L) added and stirred vigorously for 30 mins then transferred to separating funnel and organic layer separated.

Aqueous was transferred back to reaction vessel and further extracted with MTBE (2.5 L), stirred for mins vigorously then erred to separating funnel and organic layer separated. Organics were combined, dried (MgSO4), ed and concentrated to a brown oil. The oil was dissolved in pentane (500 ml) and ether (300ml). HCl (2M in ether) (434.8 mL of 2 M, 869.6 mmol) was added slowly with stirring. On complete addition the mixture was stirred for 20 mins then solid collected by tion, washed with ether and dried under vacuum for 1hr. to leave product 18 as a beige solid g, 69%); 1H NMR (500 MHz, DMSO—d6) 5 8.77 (2H, s); 19F NMR (500 MHz, DMSO—d6) 5 — 124.8; MS 210.8.

Step 2: tert—butyl I-(3-br0m0fluor0pyridinyl)piperidinecarb0xylate 26 3—bromo—4—chloro—5—fluoro—pyridine hydrochloride 18 (62 g, 251.1 mmol) was ded in DCM (600 mL) and stirred. The mixture was cooled in an ice bath and sodium hydroxide (276.2 mL of 1 M, 276.2 mmol) was added slowly. The resulting mixture was stirred for 1 hour. The mixture was phase-separated. More DCM/water was added to aid phase tion. Some tarry particulates remained in the aqueous phase. The organic phase was washed with brine, dried (MgSO4), filtered and concentrated. The residue was triturated with heptane. The e solution was filtered through a florsil pad, eluting with heptane. The filtrate was concentrated to an oil which solidified. This gave 41g of free base.

A thoroughly stirred mixture of o—4—chloro—5—fluoropyridine free base (55 g, 0.26 mol), potassium fluoride (31 g, 0.53 mol) and Me4NCl (5.8 g, 53 mmol) in DMSO (400 mL) was heated to 130°C for 2 hours. The reaction e was cooled to room temperature and tert—butyl piperidinecarboxylate hydrochloride 22 (66 g, 0.30 mol) and DIPEA (65 g, 0.50 mol) were added.

The reaction mixture was stirred at room temperature overnight. The solvent was evaporated in vacuo. The e was portioned between DCM/water. The organic layer was washed with water (3x), dried over , and filtered over silica gel using DCM as eluent. The filtrated was evaporated to give tert—butyl 1—(3 —bromo-5 -fluoropyridinyl)piperidine—4-carboxylate 26 (61 g, 65%) as a light yellow solid; 1H NMR (500 MHz, DMSO-d6) 5 8.47 (s, 1H), 8.41 (d, 1H), 3.39 — 3.36 (m, 2H), 3.12 (tt, 2H), 2.49 — 2.43 (m, 1H), 1.91 — 1.87 (m, 2H), 1.71 — 1.64 (m, 2H) and 1.43 (s, 9H); 19F NMR (500 MHz, DMSO—d6) 5 —135.2; MS (ES+) 361.0.

Step 3: utyl I-(3-aminofluor0pyridinyl)pmeridine-él-carboxylate 27 Tert—butyl romo—5—fluoropyridinyl)piperidinecarboxylate 26 (800g, 2.23mol) was dissolved in 1,4-dioxane (7.5L). Diphenylmethanimine (484g, 2.67mol) was added in one portion followed by cesium carbonate (1.45kg, 4.45mol), Xantphos (129g, 223mmol) and Pd2(dba)3 (102g, 111mmol). Additional 1,4—dioxane (2.9L) was added and the mixture heated to 95°C under nitrogen until the reaction was complete (determined by HPLC analysis). The e was cooled to °C and ethyl acetate (8L) and water (4L) were added. The organic phase was isolated and washed with water (4L) and brine (3.5L) and dried over magnesium sulphate and d. The filtrate was concentrated to a brown oil (1 .3Kg). The oil was dissolved in 2-methyltetrahydrofuran (7.2L) and 2M HCl was added at 20°C and the mixture d for 30 minutes. The aqueous layer was isolated and the organic layer extracted with 2M HCl (1.2L). The combined aqueous was neutralised with 2M NaOH (5.4L, pH 8—9). The product was extracted into 2-methyltetrahydrofuran (14L then 2x5L). The combined extracts were washed with water (1.6L) and the organic solution concentrated. The residue was slurried in acetonitrile (2L), filtered and dried. This gave the product 27 as a white solid (568.7g, 86.5%); 1H NMR (500 MHz, DMSO—dg) 5 7.82 (d, 1H), 7.63 (d, 1H), 5.22 (s, 2H), 3.11 — 3.00 (m, 2H), 2.91 (tt, 2H), 2.36 (tt, 1H), 1.88 — 1.83 (m, 2H), 1.79 — 1.71 (m, 2H), 1.43 (s, 9H); 19F NMR (500 MHz, DMSO—d6) 5 — 140.0; MS (ES+) 297.1.

Pre aration 8: S nthesis of tert-but l i eridinecarbox late CBz tBuOH, EDCI CBz H N N DMAP Pd/C 10%, H2 N DCMRT, 12h EtOAc, RT, 12h (91%) (91%) 0 OH 0 OtBu o OtBu 22 Step I: I-benzyltert—bulyl piperidine-1,4-dicarb0xylate 21 ] In a 5L flange flask was charged 1-benzyloxycarbonylpiperidinecarboxylic acid 20 (200 g, 759.6 mmol) in DCM (500.0 mL) followed by additional DCM (2.000 L), t—butanol (140.8 g, 181.7 mL, 1.899 mol) and DMAP (46.40 g, 379.8 mmol). The mixture was cooled on lt/water bath (internal -3.4°C). yliminomethyleneamino)-N,N—dimethyl-propanamine (Hydrochloric Acid (1)) (145.6 g, 759.6 mmol) was added portionwise over 15 mins, with addition funnel rinsed with DCM (500.0 mL). Mixture was stirred on ice bath for 2hr. Ice bath was then removed (internal 3°C) and allowed to warm to room temperature overnight. Mixture was washed with 5% citric acid (2 x 500 mL), then sat. NaHCO3 (500 mL), water (500 mL), and organics dried over MgSO4, which was then d and concentrated in vacuo to leave product 21 as a viscous light yellow oil which turned to a white solid on standing. (246. 1 g, 101%). 1H NMR (500 MHz, DMSO-d6) 5 7.40 — 7.31 (m, 5H), 5.08 (s, 2H), 3.90 (dt, 2H), 2.93 (br s, 2H), 2.43 (tt, 1H), 1.80 — 1.76 (m, 2H) and 1.45 — 1.37 (m, 11H).

Step 2: tert—butylpzperidinecarb0xylate 22 To a 3L flask under nitrogen was charged Pd on C, wet, Degussa , 50% water) (8.120 g, 76.30 mmol) then EtOAc (1.706 L). The mixture was degassed via Nz/vacuum cycles (3x), then a on of 1-benzyltert—butyl piperidine-1,4-dicarboxylate 21 (243.7 g, 763.0 mmol) in EtOAc (243.7 mL) was added. Mixture was stirred under a hydrogen atmosphere overnight.

Hydrogen was ished and mixture was d for a further 3.5hr. Methanol (60 mL) was added to aid ution of precipitate then filtered through celite, washing through with methanol. Filtrate concentrated in vacuo to leave a brown oil with a slight suspension of a white solid, 138.6g. Solid removed by filtration, and washed with minimal EtOAc. Filtrate was concentrated in vacuo to leave desired product as a light brown oil (129g, 91%). 1H NMR (500 MHz, DMSO—d6) 5 2.88 (dt, 2H), 2.44 (td, 2H), 2.23 (tt, 1H), 1.69 — 1.64 (m, 2H) and 1.41 — 1.33 (m, 11H).

Pre aration 9: S nthesis of 1- oxetan l i erazine oxetanone 1.2 eq NaBH(OAc)3 2 eq 9 Pd/C 10%, H2 N THF N 130323302” E j 0°C to RT, o/n E j ’ /—\ i N wN<>O o 020 100% 83/.

O O D b 25 23 24 Step I: benzyl 4-(oxetan-S-ybpiperazine-I—carboxylate 24 ] Benzyl piperazine-l-carboxylate 23 (27.3 mL, 142.2 mmol) was dissolved in dry THF (313.1 mL) and oxetan—3—one (12.29 g, 10.93 mL, 170.6 mmol) was added. The resulting solution was cooled in an ice—bath. NaBH(Oac)3 (59.99 g, 284.4 mmol) was added portionwise over 30 mins, about a quarter was added. Mixture removed from ice bath, allowed to warm to room temperature then continued adding the NaBH(Oac)3 portionwise over 30 mins. On complete addition, an exotherm from 22°C slowly to 32°C was observed, whereby the mixture was subsequently cooled on an ice bath until an internal of 22°C was reached. The ice bath was d and the reaction mixture’s internal temp was steady at 22°C. The mixture was stirred at room temperature overnight.

The resulting white suspension was quenched by addition of 2M sodium carbonate solution (approx 150 mL) (pH = 8) and concentrated under reduced pressure to remove THF. Product was then extracted with EtOAc (3 x 250 mL). Organics were combined, dried over MgSO4, filtered and concentrated under reduced pressure to leave product 24 as a white solid (32.7g 83% yield). 1H NMR (500 MHz, DMSO-d6) 5 7.39 — 7.30 (m, 5H), 5.07 (s, 2H), 4.52 (t, 2H), 4.42 (t, 2H), 3.43 — 3.39 (m, 5H) and 2.22 (t, 4H). MS (ES+) 276.8.

Step 2: I—(oxetan-S-priperazine 25 In a 1L flask was added Pd(OH)2 (1.661 g, 2.366 mmol) under nitrogen. MeOH (130.8 mL) and EtOAc (261.6 mL) were added and the mixture degassed via vacuum/nitrogen cycles (3x).

Benzyl 4-(oxetan-3 -yl)piperazine-l-carboxylate 24 (32.7 g, 1183 mmol) was then added and the mixture stirred under a hydrogen atmosphere over the d. Mixture was filtered through a pad of Celite, washing through with EtOAc then methanol. Filtrate was concentrated in vacuo to leave product 25 as an orange oil , quantitative . 1H NMR (500 MHz, DMSO—d6) 5 4.51 (t, 2H), 4.41 (t, 2H), 3.36 — 3.30 d signal, 1H), 2.69 (t, 4H) and 2.14 (br s, 4H).

Com ound 1-2 and 2-amin0fluoro-N- 5—flu0r0-4— 4- 2 2 3 3 5 5 6 deuter0-4— oxetan- N\ NH NH2 2 O / i) TFA/DCM (1/3) / N/ Pyridine \ / N \ / triethylsilane ‘N / N H 90°C 12h “ii/K0 NF/ I1 12h N N N II) 4M HCI NMP RT 20min /, \ N / </\ F :PF .HCI OtBu OtBu OH 6a* 27 29 D3“ DD NH2 NH2 0 0 /N /N ”D l ”D “(I \’ regime“: “I“ \/ o N H c)3, C I gaETEtBN F F N/ // N N Dill” \ N N N / u \ \ / / RT18h RTISII D D '_l 29 1-2 Step I: tert—bulyl I-(3-(2-amin0fluor0pyrazolo[1,5-a]pyrimidinecarb0xamid0)flu0r0pyridin- 4-yl)pzperidine—4-carb0xylate 28 A mixture of orobenzotriazolyl) 2-aminofluoro-pyrazolo[1,5-a]pyrimidine carboxylate 621* (41.69 g, 119.9 mmol) and tert—butyl 1-(3-aminofluoropyridyl)piperidine carboxylate 27 (32.2 g, 109.0 mmol) in pyridine (483 mL) was heated at 90°C for 12h. The reaction was cooled to RT, EtOH was added (322 mL), and the mixture stirred at RT for 10 mins. The solid was collected by filtration, washed well with ethanol and dried by suction to leave 28 as a yellow solid (33g, 64%).

Step 2: I-(3-(Z-aminofluor0pyrazolo[1,5-a]pyrimidine-S-carboxamido)flu0r0pyridin yl)pzperidinecarb0xylic acid 29 To a suspension of tert—butyl 1—[3—[(2—aminofluoro-pyrazolo[1,5—a]pyrimidine—3— carbonyl)amino]fluoropyridyl]piperidinecarboxylate 28 (69.7 g, 147.2 mmol) in DCM (348.5 mL) were added triethylsilane (18.83 g, 25.87 mL, 161.9 mmol) followed by TFA (151.1 g, 102.1 mL, 1.325 mol). The resulting solution was stirred at RT for 12h. The mixture was trated in vacuo to leave an orange solid which was triturated in DCM (200 mL) for 20 mins.

The solid was collected by filtration, washed with minimal DCM and dried by suction to afford the desired the trifluoroactate t as a yellow solid (75.2g, 96%).

To a solution of 1-[3-[(2-aminofluoro-pyrazolo[1,5-a]pyrimidine-3 -carbonyl)amino] fluoropyridyl]piperidinecarboxylic acid trifluoroacetate (73 g, 124.7 mmol) in NMP (662.7 mL) was added hydrogen de (4M in dioxane) (37.4 mL of 4 M, 149.6 mmol). The reaction was stirred at RT for 20 mins then the solid was collected by filtration, washed with minimal NMP then MTBE, dried by suction to afford pure t hydrochloride 29 as a light yellow solid.

Step 3: 2-amin0fluor0-N-(5-fluor0(4-(2,2, 3, 3,5, 5, 6, deuter0-piperazine-I - carbonprzperidin-I-yl)pyridyl)pyrazolo[1,5-a]pyrimidine-S-carboxamide (Compound I-2) (Benzotriazolyloxy-dimethylamino-methylene)-dimethyl-ammonium trifluoroborate (127.3 mg, 0.3966 mmol) was added to a mixture of 1—[3—[(2-amino—6—fluoro—pyrazolo[1,5— a]pyrimidinecarbonyl)amino]fluoropyridyl]piperidinecarboxylic acid hydrochloride 29 (150 mg, 0.3305 mmol) 1.652 mmol) and Et3N , 2,2,3,3,5,5,6,6—octadeuteriopiperazine (155.6 mg, (83.6 mg, 115.2 uL, 0.8262 mmol) in DMF (5 mL). The reaction mixture was stirred at RT for 18h.

The crude mixture was purified by preparative HPLC to afford I-2 as a white solid (114mg, 48%).

Step 4: 2-amin0fluor0-N-(5-fluor0(4-(2,2, 3,3, 5, 5, 6, 6-octadeuter0(oxetan-S-przperazine-I - carbonprzperidin-I-yl)pyridineyl)pyrazolo[1,5-a]pyrimidine-S-carboxamide (Compound I-3) Sodium triacetoxyborohydride (24.67 mg, 0.1164 mmol) was added to a on of oxetan—3—one (7.271 mg, 0.1009 mmol), 2—amino—6—fluoro—N—(5—fluoro—4—(4—(2,2,3,3,5,5,6,6— WO 85132 octadeutero-piperazinecarbonyl)piperidinyl)pyridine-3 -yl)pyrazolo[1,5 -a]pyrimidine-3 - carboxamide 13 (56 mg, 0.07761 mmol) and acetic acid (13.98 mg, 13.24 uL, 0.2328 mmol) in DMF (2 mL). The on mixture was stirred at RT for 18h. The solution was quenched with methanol and water and the crude mixture was purified by preparative HPLC to afford the desired product I-3 (20mg, 46%). 1H NMR (500 MHz, DMSO—d6) 5 10.64 (s, 1H), 9.67 (s, 1H), 9.48 (dd, 1H), 9.26 (dd, 1H), 8.26 (d, 1H), 6.79 (s, 2H), 4.55 (t, 2H), 4.47 (t, 2H), 3.63 (m, 1H), 3.20 (m, 2H), 3.15 (m, 2H), 2.95 (m, 1H), 2.10 (m, 2H), 1.74 (d, 2H); ES+ 550.4.

Compound Analytical Data H NMR (500 MHz, methanol-d4) 5 1.87 (2H, m), 2.27- 2.33 (2H, m), 2.55 (4H, m), 2.97—3.03 (1H, m), 3.18 (2H, m), 3.70—3.85 (4H, m), 4.67—4.70 (2H, m), 4.75—4.78 (2H, m), 8.16 (1H, d), 9.00 (1H, dd), 9.17 (1H, dd), 9.68 (1H, s), .65 (1H, s). 1H NMR (500 MHz, DMSO—d6) 5 10.64 (s, 1H), 9.67 (s, 1H), 9.48 (dd, 1H), 9.26 (dd, 1H), 8.26 (d, 1H), 6.79 (s, 2H), 3.20—3.25 (m, 2H), 3.05—3.07 (m, 2H), 2.95-2.98 (m, 1H), .12 (m, 2H), 1.74 (d, 2H). 1H NMR (500 MHz, DMSO—d6) 5 10.64 (s, 1H), 9.67 (s, 1H), 9.48 (dd, 1H), 9.26 (dd, 1H), 8.26 (d, 1H), 6.79 (s, 2H), 4.55 (t, 2H), 4.47 (t, 2H), 3.63 (m, 1H), 3.20 (m, 2H), 3.15 (m, 2H), 2.95 (m, 1H), 2.10 (m, 2H), 1.74 (d, 2H).

Solid Forms of nd I-1 Compound I-1 has been prepared in s solid forms, including salts, solvates, hydrates, and anhydrous forms. The solid forms of the t ion are useful in the manufacture of medicaments for the treatment of cancer. One embodiment provides use of a solid form described herein for treating cancer. In some embodiments, the cancer is triple negative breast cancer, pancreatic cancer, small cell lung cancer, colorectal cancer, ovarian cancer, or non-small cell lung cancer. Another embodiment provides a pharmaceutical composition comprising a solid form described herein and a pharmaceutically acceptable carrier.

WO 85132 Applicants describe herein a plurality of novel solid forms of Compound I-1. The names and stoichiometry for each of these solid forms are provided in Table 2 below: Table2 Examole 5 0 ethanol solvate 1:0.72 Exam-166a 1:45 Examle 6b ——————— Exam-167 N/A Exam-168 ——————— Exam-169 N/A Exam-1610 N/A Exam-1611 1:1 Exam-1612 1:13 Exam-1613 1:0.44 Example 14 Compound I isopropanol solvate 1:0.35 ssNMR Experimental Method Solid state NMR spectra were acquired on the Bruker—Biospin 400 MHz Advance III wide—bore spectrometer equipped with Bruker—Biospin 4mm HFX probe. Samples were packed into 4mm ZrOz rotors (approximately 70mg or less, depending on sample availability). Magic angle spinning (MAS) speed of typically 12.5 kHz was applied. The temperature of the probe head was set to 275K to minimize the effect of frictional heating during spinning. The proton relaxation time was ed using 1H MAS T1 saturation recovery relaxation experiment in order to set up proper e delay of the 13C cross-polarization (CP) MAS experiment. The recycle delay of 13C CPMAS experiment was adjusted to be at least 1.2 times longer than the measured 1H T1 relaxation time in order to maximize the carbon spectrum signal-to-noise ratio. The CP contact time of 13C CPMAS experiment was set to 2 ms. A CP proton pulse with linear ramp (from 50% to 100%) was ed. The nn-Hahn match was optimized on external reference sample ne).

Fluorine spectra were acquired using proton led MAS setup with recycled delay set to approximately 5 times of the measured 19F T1 relaxation time The fluorine relaxation time was measured using proton decoupled 19F MAS T1 saturation recovery relaxation experiment. Both carbon and fluorine spectra were acquired with SPINAL 64 decoupling was used with the field strength of approximately 100 kHz. The chemical shift was referenced against external standard of adamantane with its upfield resonance set to 29.5 ppm.

Example 5: Compound I-1 [ethanol e] Compound I-1 ethanol solvate can be prepared according to the s described in Example 1, Step 4.

XRPD of Compound I-1 {ethanol solvate) The XRPD n of Compound I-1° ethanol solvate was recorded at room temperature in reflection mode using a PANalyticaZ diffractometer equipped with an Empyrean tube source and a PIXcel 1D detector yticaZ, The Netherlands). The X-ray tor was operating at a voltage of 45 kV and a current of 40 mA. The powder sample was placed in a silicon holder. The data were over the range of 3°-39° 2 theta with a step size of 0.0130 and a dwell time of 121s per step. Figure la shows the X-ray powder diffractogram of the sample which is characteristic of crystalline drug substance.

Table 3a depicts representative XRPD peaks from Compound I-1° ethanol solvate: Table 3a: Representative XRPD Peaks 2 12.7 6.4 3 13.6 12.2 4 14.3 7.5 14.9 9.5 7 16.2 15.7 8* 50.6 * 19.7 35.3 11 25.2 12 23-1 13 3.5 22.8 11.2 16 18.2 18 23.8 91.4 * 24.4 72.8 21 15.1 23 26.3 6.0 24 27.5 5.8 * 29.0 44.9 26 30.0 9.7 NN 00\l 30.9 4.6 31.5 4.5 Thermo Analysis of Compound I-1 ol solvate) A thermal gravimetric analysis of Compound I-1° ethanol solvate was med to determine the t weight loss as a function of temperature using the Discovery TGA (TA Instruments Trios). A sample (8.338 mg) was added to a pre-tared aluminum pan and heated from ambient temperature to 310°C at 20°C/min. The TGA results seen in Figure 2a show a large weight loss of 5.76% between 166°C (onset) and 219°C (end point). This weight loss corresponds to approximately 0.72 molar equivalents of ethanol. The subsequent weight loss seen at 290°C is a result of g/degradation.

Differential ng Calorimetg of Compound I-1 {ethanol solvate) Differential scanning calorimetry of Compound I-1° ethanol solvate was measured using the TA Instrument DSC Q2000. A sample (1.84 mg) was weighed in a pre—punched pinhole um hermetic pan and heated from ambient temperature to 300°C at 20°C/min. The DSC results seen in Figure 3a show a ation endotherm at 169 °C (onset) followed by a single melting endotherm at 258°C (onset). ssNMR of nd I-1 {ethanol solvate) A solid state 13C NMR spectrum of Compound I ethanol solvate is shown in Figure 4a.

Table 3b provides chemical shifts of the relevant peaks.

Table 3b: Solid State 13C NMRspectrum of Compound I-1 (ethanol solvate) Peak # Intensity 1* 175.4 53.9 2 162.4 58.4 3 160.0 14.1 4 157.4 150.7 6 148.2 7 145.8 8 140.1 9* 138.0 136.1 11 134.3 12* 123.1 13 89.0 14 76.8 76.1 16* 57.8 17 51.6 18 48.9 19* 44.0 42.2 21 38.8 22 30.9 23 28.7 24* 19.5 A solid state 19F NMR um of Compound Iethanol solvate is shown in Figure 5a.

Table 3c provides chemical shifts of the relevant peaks.

Table 3c: Solid State 19F NMR Spectrum of Compund I-1 (ethanol solvate) 436-0 451-6 Example 6a: Compound I-1 [hydrate I] Compound I-1° ethanol e (lOOOmg), prepared according to the methods described in Example 1, Step 4, was slurried in water (20mL) for 4 days at room temperature. The suspension was centrifuged and the residual solids were isolated then dried overnight in a 35°C vacuum oven to afford nd I e I as a yellow powder.

XRPD of Compound I-1 [hydrate I] The XRPD pattern of Compound I-1° hydrate I was recorded at room temperature in reflection mode using a Bruker D8 Discover diffractometer equipped with a sealed tube source and a Hi-Star area detector (Bruker AXS, Madison, WI, Asset V012842). The X-ray generator was operating at a voltage of 40 kV and a current of 35 mA. The powder sample was placed in a nickel holder. Two frames were registered with an exposure time of 120 seconds each. The data were subsequently integrated over the range of 3.5°—3 9° 2—theta with a step size of 0.020 and merged into one continuous pattern. Figure lb shows the X-ray powder diffractogram of the sample which is teristic of crystalline drug substance.

Table 4a s representative XRPD peaks from nd I-1° hydrate I: Table 4a: Representative XRPD Peaks 1 1.4 2 3.0 3 7.8 4 100.0 * 51.0 6 3.0 7 10.5 8 10.4 10.8 9 11.2 5.9 11.5 8.7 11 11.5 12* 16.0 13* 10.9 14 7.2 9.2 16 10.5 17 14.8 18* 10.8 19 14.1 11.6 21 10.2 22 4.5 23 7.8 24* 13.6 10.9 26 4.9 27 32.3 2.2 Thermo Analysis of Compound I-1 ghydrate I) ] A thermal gravimetric analysis of Compound I-1° hydrate I was performed to determine the percent weight loss as a function of time using the TA Instrument TGA Q5000 (Asset V014258).

A sample (7.380 mg) was added to a pre-tared aluminum pan and heated from ambient temperature to 350°C at 10°C/min. The TGA results seen in Figure 2b show a large initial weight loss up to WO 85132 2014/068713 100°C ed by a small amount of additional weight loss prior to melting/degradation. The initial weight loss of 14.56% corresponds to imately 4.5 molar equivalents of water. The onset temperature of melting/degradation is 292°C. ential Scanning Calorimetg of Compound I-1 [hydrate I] Differential scanning calorimetry of Compound I-1° hydrate I was measured using the TA Instrument DSC Q200 (Asset V005642). A sample (5.598 mg) was d in a pre—punched pinhole aluminum hermetic pan and heated from ambient ature to 350°C at 10°C/min. The DSC results seen in Figure 3b show an initial broad endothermic event that corresponds to de— hydration and subsequent melting to an amorphous form. Following the melt there is a Tg at 125°C, stallization at 180°C, a melt at 257°C, then a final melt/degradation event at 278°C.

Example 6b: Compound I-1 [hydrate II] Compound I-1° ethanol solvate (1000mg), prepared according to the methods described in Example 1, Step 4, was slurried in water (20mL) for 4 days at room temperature. The suspension was centrifuged and the residual solids were isolated to afford Compound I hydrate II as a yellow paste.

XRPD of Compound I-1 [hydrate II] The XRPD pattern of Compound I-1°hydrate II was recorded at room temperature in reflection mode using a Bruker D8 Discover diffractometer equipped with a sealed tube source and a Hi-Star area detector (Bruker AXS, Madison, WI, Asset V012842). The X-ray generator was ing at a voltage of 40 kV and a current of 35 mA. The powder sample was placed in a nickel holder. Two frames were registered with an exposure time of 120 seconds each. The data were subsequently integrated over the range of 3.5°-39O 2—theta with a step size of 0.020 and merged into one continuous pattern. Figure 4b shows the X-ray powder diffractogram of the sample which is characteristic of crystalline drug substance.

Table 4b depicts representative XRPD peaks from Compound I-1° hydrate II: Table 4b: Representative XRPD Peaks 8* 11.9 29.9 9 45.6 25.2 11 22.2 12 19.7 13 17.2 14 39.7 21.1 16* 24.7 17 10.3 18 28.6 19 12.0 8.8 21* 13.0 22 12.6 23 5.3 24 12.2 9.5 26 9.2 27 14.9 ssNMR of Compound I-1 {hydrate 111 A solid state 13C NMR spectrum of Compoun I-1° hydrate II is shown in Figure 5b. Table 40 provides Chemical shifts of the relevant peaks.

Table 4c: Solid State 13C ctrum of Compoun I-1 (hydrate II) Intensity 49.2 24.9 3 161.3 4 160.9 159.7 6* 158-2 7 151-9 8 149-1 9* 142-9 136-3 12 132-9 40.1 13 130.5 14 122-8 * 85-1 16 76-9 17 76.4 18* 58.9 19 50.2 49.5 21 48.5 22 45.0 23 41 .8 24 37.2 * 31 .9 26 28.9 ] A solid state 19F NMR spectrum of Compound drate II is shown in Figure 6b. Table 4d provides Chemical shifts of the relevant peaks.

Table 4d: Solid State 19F NMR um of Compoun I-1° hydrate II Intensity 1* -138.0 8.2 2* -152.7 12.5 Example 7: Compound I-1 [anhydrous form A] Compound I-1° ethanol solvate (lOOOmg), prepared according to the methods described in Example 1, Step 4, was slurried in THF (20mL) for 72hr at room temperature. The suspension was fuged and the residual solids were isolated then dried overnight in a 35°C vacuum oven to afford compound I anhydrous form A (“form A”) as a yellow powder.

In an alternative process, compound I-1° amorphous form (15. lg; 0.028mol), prepared according to the method in Example 2, step 3, was suspended in a mixture of 2-propanol (300mL) and water (lOOmL). The mixture was stirred and heated to 70-75°C and filtered whilst hot. The ing clear te was heated and distilled and solvent replaced with anol until the contents temperature reached 825°C. The resulting suspension was cooled to 15°C over 10 hours and stirred for a further 5 hours. The solids were collected by filtration, dried by suction for 1 hour then dried in a vacuum oven for 20 hours at 60°C to give compound I-1° anhydrous form A ; 92%).

A number of other solvents may be utilized to prepare compound I anhydrous form A.

Table 5a below summarizes the methods.

Table 5a: Solvents Used to Prepare Form A Anisole Slurry 2—Butanone Slurry Slurry Ethyl acetate Slurry Heptane Hot Slurry Isopropanol Slurry Isopropyl acetate TBME Slurry THF Slurry XRPD of Compound I-1 ganhydrous form A1 ] The XRPD pattern of Compound I-1° anhydrous form A was recorded at room temperature in reflection mode using a Bruker D8 Discover diffractometer equipped with a sealed tube source and a Hi—Star area detector (Bruker AXS, Madison, WI, Asset V012842). The X—ray generator was operating at a e of 40 kV and a current of 35 mA. The powder sample was placed in a nickel holder. Two frames were registered with an exposure time of 120 seconds each.

The data were subsequently integrated over the range of 3.50—3 9° 2—theta with a step size of 0.020 and merged into one continuous pattern. Figure 1c shows the X-ray powder ctogram of the sample which is characteristic of crystalline drug substance.

Table 5b depicts representative XRPD peaks form Compound I anhydrous form A: Table 5b: Representative XRPD Peaks WO 85132 18* 31.8 16.0 Thermo Analysis of Compound I-1 (anhydrous form A) A thermal gravimetric analysis of Compound I-1° anhydrous form A was performed to determine the percent weight loss as a function of time using the TA ment TGA Q5000 (Asset V01425 8). A sample (7.377 mg) was added to a pre-tared aluminum pan and heated from ambient temperature to 350°C at 10°C/min. The TGA results seen in Figure 2c show very little observed weight loss prior to melting or thermal degradation. From ambient temperature to 265°C, the weight loss is 0.96%. The onset temperature of degradation is 292°C.

Differential Scanning Calorimetfl of Compound I-1 rous form A) Differential scanning calorimetry of Compound I-1° anhydrous form A was ed using the TA Instrument DSC Q2000 (Asset V014259). A sample (3.412 mg) was weighed in a prepunched pinhole aluminum hermetic pan and heated from ambient ature to 350°C at in. The DSC results seen in Figure 3c show a single endothermic melting event at 262°C.

There are two distinct peaks contained within the melting event which are separated by about 1°C.

Composition and Preparation ofactive Tablets Containing Anhydrous Form A Composition of Form A 10 mg tablet The formulation compositions for both the dry granulation and tablet blends of the active Form A 10 mg tablets are described in Tables 5c and 5d. The overall composition specification of the tablets is bed in Table 5e.

Table 5c: Form A (10mg) Intragranular Blending Form A 10.00 10.26 Lactose Monoh drate, #316, NF, PhEur, JP 27.50 28.20 Avicel PH- 101 (microcrystalline cellulose), 55.00 56.41 NF, PhEur, JP Ac—Di—Sol (croscarmellose sodium), NF, PhEur, JP Sodium Stea lFumarate, NF, PhEur, JP 2.00 2.05 97.50 100.00 Table 5d: Form A (10mg) Tablet Composition Form A Intragranular Blend (Milled) Ac—Di—Sol (croscarmellose sodium), NF, PhEur, JP Sodium Stearyl Fumarate, NF, PhEur, JP Total Table 5e: Form A (10mg) Tablet Overall Composition Form A e Monohydrate, #316, NF, PhEur, JP intra Avicel PH-101, NF, PhEur, JP granular Ac-Di-Sol, NF, PhEur, JP Sodium Stea lFumarate, NF, PhEur, JP _ranules: extra Ac-Di-Sol, NF, PhEur, JP 1.50 granular Sodium Stearyl Fumarate, NF, PhEur, JP 1.00 total core : 100.00 Composition of Form A 50 mg tablet The formulation itions for both the dry granulation and tablet blends of the active Form A 50 mg tablets are described in Tables 5f and 5g. The overall composition specification of the tablets is described in Table 5h.

Table 5f: Form A (50mg) Intragranular Blending e Monoh drate, #316, NF, PhEur, JP 137.50 28.20 Avicel PH- 101 (microcrystalline cellulose), 275.00 56.41 NF, PhEur, JP Ac—Di—Sol '00 croscarmellose sodium JP , NF, PhEur, Sodium l Fumarate, NF, PhEur, JP 10.00 2.05 Total 487.50 100.00 Table 5g: Form A (50mg) Tablet Composition Form A lntragranular Blend 487.50 97.50 Milled Ac-Di-Sol 1.50 (croscarmellose sodium), NF, PhEur, JP Sodium Stearyl Fumarate, 1 00 NF, PhEur, JP ‘ Total 100.00 Table 5h: Form A (50mg) Tablet Overall Composition e Monohydrate, #316, NF, PhEur, JP intra AVicel PH-101, NF, PhEur, JP granular Ac-Di-Sol, NF, PhEur, JP Sodium Stearyl Fumarate, NF, PhEur, JP total _ranules: extra Ac-Di-Sol, NF, PhEur, JP 1.50 granular Sodium Stearyl Fumarate, NF, PhEur, JP 1.00 total core tablet: 100.00 Process for Preparing Form A 10 mg and 50 mg Tablets Step I. Pre-Granulation Mixing: Form A was passed through a cone mill assembled with a 24R round holed screen and a rounded edge type impeller at an impeller rate of 1500 rpms. e monohydrate, microcrystalline cellulose, and intra—granular croscarmellose sodium were screened through a #30 mesh sieve. The cone milled Form A and all the screened components were then blended for 10 minutes at 26 rpm.

Sodium l fumarate was hand sieved through a 60 mesh screen and then charged into the r and blended with the materials for 3 minutes at 26 rpm. Samples were pulled for blend mity analysis.

Step II. Dry Granulation: The blend was dry granulated on a Gerteis Minipactor. The blend was passed through the roller compactor, led with a combination of smooth faced and knurled faced compaction rolls, at a 2 rpm roll speed with 5KN/cm roll force and a 2 mm roll gap. Compacted powder was then granulated with a pocketed type milling roll through a 1 mm screen with 80 rpm mill speed.

Step III. Final Blending: Extra—granular croscarmellose sodium and sodium stearyl te were hand sieved through 30 and 60 mesh screens, tively. granular croscarmellose sodium was blended with the dry granulate for 5 minutes at 32 rpm. Extra-granular sodium stearyl fumarate was then added to the bulk mixture and mixed for 3 s at 32 rpm. Samples were pulled for blend uniformity analysis. The blend was sealed in double Low Density Polyethylene bags within a hard secondary container to protect from puncture.

Step IV. Tablet Compression: ] A tablet compression machine (Piccola D—8 Rotatory Press) was partially tooled (2 stations out of 8 stations) with a 0.25” standard round concave tooling for 10 mg strength and 0.568” x 0.2885” caplet tooling for 50 mg th. Turret speed was 25—35 rpm. The in—process control testing for tablets included average weight, individual weight, and hardness, as shown in Table 5i.

Table 5i: Form A (10mg and 50 mg) Tablet Compression In-process Control Specifications we1ht m Cgstal Preparation of Form A ] Form A was crystallized from a DCM/ heptane mixture by slow evaporation of the solvents. A colorless needle shaped crystal with dimensions 0.10 x 0.02 x 0.02 mm was chosen for the diffraction experiment on a Bruker APEX ll CCD ctometer with Cu Ka radiation at room temperature. The structure was solved by direct methods and refined by the SHELXTL package.

Form A Crystal Experimental: ] The crystal shows monoclinic cell with P21/c centrosymmetric space group. The lattice parameters are a = 15.29(3)A, b = 12.17(2)A, c= 14.48(3)A, or = 90°, E = 107.22(3)°, y = 90°, volume = 2573(9)A3. The refinement gave the R factor of 6.9%. Conformational plots of Compound I-1° anhydrous form A based on single crystal X-ray analyses are shown in Figures 4c and 5c.

Compound I-1° anhydrous form A appears d in the asymmetric unit (Figure 4c). As shown in Figure 5c, Compound I-1° ous form A les form a one-dimensional chain along the b- axis that is stabilized by the inter—molecular hydrogen bonds between the amine and ne groups.

Multiple chains stack in three dimensions with approximately 4.3A inter-layer spacing.

Table 5j: Crystal data for Form A C25H29F2N903 Z = 4 Mr=541.57 F(000)= 1136 Monoclinic, P21/c Dx = 1.398 Mg m'3 a = 15.29 (3) A Cu K01 radiation, 7» = 1.54178 A b=12-17(2)A u=0.89 mm'l c= 14.48 (3)A T=296K [3 = 107.22 (3)0 Needle, colorless V= 2573 (9) A3 0.10 X 0.02 X 0.02 mm Geometry: All esds (except the esd in the dihedral angle between two l.s. ) are estimated using the full covariance matrix. The cell esds are taken into account individually in the estimation of esds in distances, angles and torsion angles; ations between esds in cell parameters are only used when they are defined by crystal symmetry. An approximate (isotropic) treatment of cell esds is used for estimating esds involving ls. .

Table 5k: Data collection parameters for Form A crystal Bruker APEX II CCD Rim = 0.084 diffractometer Radiation source: sealed tube Gmax = 53.60, 0min = 3.00 oscillation photos around 00 and (1) scans h — —15—>15 9104 measured reflections k = —12—> 1 1 293 9 independent reflections Z = —1 1—> 14 Data collection: Apex 11; cell refinement: Apex II; data reduction: Apex II; program(s) used to solve structure: SHELXS97 (Sheldrick, 1990); program(s) used to refine ure: SHELXL97 rick, 1997); molecular graphics: Mercury; software used to prepare al for publication: publCIF.

Table 5m: Refinement parameters for Form A crystal Least-squares matrix: full en site location: inferred from neighbouring sites Rw2>2o<F2>1=oow more) = 0.179 w = 10020002) -— (007431021 where P = (F02 -- 3 S = 0.94 (A/o)max < 0.001 2939 refiectlons A>max = 0.23 e A 3 352 parameters A>min = —0.26 e A'3 ent: Refinement of F2 against ALL reflections. The weighted R-factor wR and goodness of fit S are based on F2, conventional R—factors R are based on F, with F set to zero for negative F2. The threshold expression of F2 > 2sigma(F2) is used only for calculating R-factors(gt) etc. and is not relevant to the choice of reflections for refinement. R-factors based on F2 are statistically about twice as large as those based on F, and R-factors based on ALL data will be even larger. ssNMR of Compound I-1 [anhydrous form A] ] A solid state C NMR spectrum of Compoun I anhydrous form A is shown in Figure 6c. Table 5n provides chemical shifts of the relevant peaks.

Table 5n: Solid State 13C NMRspectrum of Form A Intensity 67.9 46.9 59.1 18.1 9 139.8 * 138.9 11 135.8 12 134.3 13 122.6 14 89.3 76.2 16* 74.1 17 59.8 18 51.7 77.2 19 50.3 98.8 49.4 91.4 21* 42.8 100.0 22 38.4 97.7 23* 31.5 84.2 24 28.3 85.4 A solid state 19F NMR spectrum of Compound Ianhydrous form A is shown in Figure 7c. Table 5p provides chemical shifts of the nt peaks.

Table 5p: Solid State 19F NMR Spectrum of Form A O Om 436.8 .1. 455.7 Example 8: Compound I-1 [anhydrous form B] Charged Compound I-1° amorphous (3.50g), prepared according to the methods described in Example 2, Step 3, was placed in a 250mL 3-neck flask, THF (70mL) was added, and agitated using an overhead stirrer at ambient temperature overnight (e.g., at least 12 hr.). The suspension was filtered under vacuum (4.25cm diameter Whatman filter paper), washed with THF (7mL), and pulled under vacuum for about 35 minutes to give a fairly hard yellow solid (2.25 g). Dried the solids under vacuum with a good bleed at 35°C ght affording 1.92 lg of nd I anhydrous form B as a yellow solid.

XRPD of nd I-1 [anhydrous form B] The XRPD pattern of Compound I anhydrous form B was recorded at room temperature in reflection mode using a Bruker D8 Discover diffractometer equipped with a sealed WO 85132 tube source and a Hi—Star area detector (Bruker AXS, n, WI, Asset V012842). The X—ray generator was operating at a voltage of 40 kV and a current of 35 mA. The powder sample was placed in a nickel holder. Two frames were registered with an exposure time of 300 seconds each.

The data were subsequently integrated over the range of 3.50—3 9° 2—theta with a step size of 0.020 and merged into one continuous pattern. Figure 1d shows the X-ray powder diffractogram of the sample which is teristic of crystalline drug substance.

Table 6a s representative XRPD peaks form Compound I anhydrous form B: Table 6a: Representative XRPD Peaks 00 UL) Thermo Analysis of Compound I-1 {anhydrous form B) A thermal gravimetric analysis of Compound I-1° anhydrous form B was med to determine the percent weight loss as a function of time using the TA Instrument TGA Q500 (Asset V014840). A sample (2.728 mg) was added to a pre-tared platinum pan and heated from ambient temperature to 350°C at 10°C/min. The TGA results seen in Figure 2d show two distinct weight loss 2014/068713 events ng 2.5% up to 175°C. The onset temperature of melting/degradation is 284°C. ential Scanning Calorimetgg of Compound I-1 (anhydrous form B) Differential scanning calorimetry of Compound I-1° anhydrous form B was measured using the TA Instrument DSC Q2000 (Asset V0123 90). A sample (2.125 mg) was weighed in a pre— d pinhole aluminum hermetic pan and heated from 30°C to 350°C at 3°C/min, modulating :: 1°C every 60 seconds. The DSC results seen in Figure 3d show an exothermic event at 177°C (likely a slight re-arrangement of the crystal structure), an endothermic melt at 257°C, re-crystallization at 8°C, then a final melt/degradation event at 280°C. ssNMR Compound I-1 {anhydrous form B1 A solid state 13C NMR spectrum of Compoun I-1°anhydrous form B is shown in Figure 4d. Table 6b provides chemical shifts of the relevant peaks.

Table 6b: Solid State 13C NMR spectrum of Form B Peak # Intensity 1* 173.4 2* 164.5 3 162.3 4 159.9 157.4 6 151.8 7 149.5 8 144.9 9 141.6 136.3 11* 133-5 12* 130.8 13 124.4 14 86.9 74.9 16 72.0 17* 67.7 18 59.5 19 50.8 * 45.3 21 40.4 22 37.9 mm. 23 30.3 24* 25.9 A solid state 19F NMR um of nd Ianhydrous form B is shown in Figure 5d. Table 6c provides chemical shifts of the relevant peaks.

Table 6c: Solid State 19F NMR Spectrum of Form B Intensity Example 9: Compound I-1 [anhydrous form C] Compound I-1° ous form B (~15mg), prepared according to the method described in Example 8, was added to nched pinhole aluminum hermetic pans and heated via DSC to 265°C at a rate of 5°C/min (3 pans, ~5mg each) to afford compound I-1° anhydrous form C as a dark yellow powder.

XRPD of Compound I-1 [anhydrous form C] The XRPD pattern of nd I anhydrous form C was recorded at room temperature in reflection mode using a Bruker D8 Discover diffractometer equipped with a sealed tube source and a Hi—Star area detector (Bruker AXS, Madison, WI, Asset V012842). The X—ray generator was operating at a e of 40 kV and a current of 35 mA. The powder sample was placed in a nickel holder. Two frames were registered with an re time of 120 seconds each.

The data were subsequently integrated over the range of 3.50—3 9° 2—theta with a step size of 0.020 and merged into one continuous pattern. Figure 1e shows the X-ray powder diffractogram of the sample which is characteristic of crystalline drug substance.

Table 7a depicts representative XRPD peaks form Compound I anhydrous form C: Table 7a: Representative XRPD Peaks 8* 13.4 29.1 9 14.2 51.8 Thermo Analysis of Compound I-1 ganhydrous form C1 A thermal etric analysis of Compound I-1° anhydrous form C was performed to determine the percent weight loss as a function of time using the TA Instrument TGA Q500 (Asset V014840). A sample (3.363 mg) was added to a pre—tared platinum pan and heated from ambient temperature to 350°C at 10°C/min. The TGA results seen in Figure 2e show no distinct weight loss events prior to melting/degradation. The onset temperature of melting/degradation is 292°C.

Differential Scanning Calorimetg of Compound I-1 {anhydrous form C) Differential scanning calorimetry of Compound I-1° anhydrous form C was measured using the TA Instrument DSC Q2000 (Asset V0123 90). A sample (4.100 mg) was d in a pre— punched pinhole aluminum hermetic pan and heated from 30°C to 350°C at 3°C/min, ting :: 1°C every 60 seconds. The DSC results seen in Figure 3e show a single endothermic melting/degradation event at 281°C. ssNMR A solid state C NMR spectrum of Compoun I-1°anhydrous form C form is shown in Figure 4e. Table 7b es chemical shifts of the relevant peaks.

Table 7b: Solid State 13C NMRspectrum of Form C Peak# 1* 175.2 2 163.3 3 162.1 4 158.2 152.4 6 149.9 7 144.9 8* 142.5 9 137.9 135.7 11* 129.6 12 123-6 13 86.5 14 76.6 * 73.5 16 59.6 17* 54.0 18 51.2 19 49.7 * 46.7 21 42.3 22 37.2 23 31.4 24 28.9 ] A solid state 19F NMR spectrum of Compound Ianhydrous form C is shown in Figure 5e. Table 7c provides chemical shifts of the relevant peaks.

Table 7c: Solid State 19F NMR Spectrum of Form C Intensity 1* -131.2 4.8 2* -150.7 12.5 Example 10: Compound I-1 [amorphous form] Compound I-1° amorphous form was prepared according to the s described in e 2, Step 3, or in Example 3, Step 3, above.

XRPD of Compound I-1 [amomhous form] The XRPD pattern of Compound I-1° amorphous form was recorded at room temperature WO 85132 2014/068713 in ion mode using a PANalyticaZ diffractometer equipped with an Empyrean Cu tube source and a PIXcel 1D detector (PANaZyticaZ, The Netherlands). The X-ray generator was ing at a voltage of 45 kV and a current of 40 mA. The powder sample was placed in a silicon holder. The data were over the range of 3°—3 9° 2 theta with a step size of 0.013° and a dwell time of 0.5s per step.

Figure 1f shows the X-ray powder diffractogram of the sample which is characteristic of amorphous drug nce.

Differential Scanning Calorimetg of Compound I-1 {amorphous form) Differential scanning calorimetry of Compound I-1° amorphous form was measured using the TA ment DSC Q2000. A sample (2.61 mg) was weighed in an aluminum non-hermetic pan and heated using the modulated mode from ambient temperature to 350°C at a heating rate of 2°C /min, with a modulation amplitude of +/—0.5°C and a period of 60s. The DSC results seen in Figure 2f show a glass transition (Tg) at 128°C (onset) with heat capacity change of 0.3 J/(g.°C). Glass transition was followed by a crystallization exotherm at 174°C (onset), which was in turn followed by a melt/ degradation event at 250°C. ssNMR of Compound I-1 (amorphous) A solid state C NMR spectrum of Compoun I amorphous form is shown in Figure 3f Table 8a provides chemical shifts of the relevant peaks.

Table 8a: Solid State 13C NMRspectrum of amorphous form IntenSIty 1* 173-8 2 162-3 3 157.6 4 149.3 * 144.2 6 134-7 7 123-1 8* 87.5 9 75.6 59.4 11 50.8 12* 45-6 13 41-8 14 38.3 * 29-5 A solid state 19F NMR spectrum of Compound orphous is shown in Figure 4f.

Table 8b es chemical shifts of the relevant peaks.

Table 8b: Solid State 19F NMR Spectrum of ous form Intensity 1 .0* 9.8 12.5 Example 11: Compound I-1 (DMSO solvate) Compound I-1° anhydrous form A (10.0g; 18.47mmol), prepared according to the methods described in Example 7, was suspended in DMSO (200mL) and heated to 55°C. The e was filtered whilst hot. The hot filtrate was stirred in a clean flask and cooled to 20—25°C then stirred for an additional 2 hours. The solids were collected by filtration, washed with DMSO (10mL), dried by suction then dried in a vacuum oven for 14 hours at 40—45°C to give compound I- 1- DMSO solvate (7.23g; 63%). 1H NMR (500 MHz, DMSO—d6) 5 10.63 (s, 1H), 9.66 (s, 1H), 9.47 (dd, 1H), 9.24 (dd, 1H), 8.24 (d, 1H), 6.78 (s, 2H), 4.54 (t, 2H), 4.46 (t, 2H), 3.60 (dt, 4H), 3.43 (m, 1H), 3.18 (m, 2H), 2.97 (m, 3H), 2.54 (s, 6H), 2.26 (dt, 4H), 2.12 (qd, 2H), 1.73 (d, 2H); 19F NMR (500 MHz, DMSO—d6) 5 —136.1, —152.8.

XRPD of Compound I-1 (DMSO solvate) The XRPD pattern of compound I-1° DMSO solvate was recorded at room temperature in reflection mode using a PANalyticaZ diffractometer ed with an Empyrean tube source and a PIXcel 1D detector (PANaZyticaZ, The Netherlands). The X-ray generator was operating at a voltage of 45 kV and a current of 40 mA. The powder sample was placed in a silicon . The data was recorded over the range of 30—390 2 theta with a step size of 0.0130 and a dwell time of 121s per step. Figure 1 g shows the X-ray powder diffractogram of the sample which is characteristic of crystalline drug nce.

Table 9 depicts representative XRPD peaks form nd I DMSO solvate: Table 9: Representative XRPD Peaks 7.0034 8.9204 .4007 WO 85132 12.4735 6.81 12.7962 12.32 13.3976 —12.25 14.8102 —29.16 .439 —15.1 .7477 —14.37 16.5454 82.57 —15.34 —20.25 —29.71 —3.68 .1178 .9143 21.3593 22.1801 23.3866 23.8312 22 24.5088 —15.65 —11.53 —8.95 —5.69 —8.63 —6.42 —8.71 33.2165 34.1902 34.6067 .45 36.5669 38.6972 Thermo Analysis of Compound I-1 {DMSO solvate) A thermal graVimetric analysis of compound I-1° DMSO solvate was performed to determine the t weight loss as a function of temperature using the Discovery TGA (TA Instruments . A sample (3.26 mg) was added to a pre-tared aluminum pan and heated from ambient temperature to 350°C at 10°C/min. The TGA results seen in Figure 2g show a large weight loss of 12.44% n 146°C (onset) and 156°C (end point). This weight loss corresponds to approximately 1 molar equivalents of DMSO. A second weight loss of 0.52% was then seen between 254°C (onset) and 262°C (end point). The subsequent weight loss seen at 304°C is a result of melting/degradation.

Differential Scanning Calorimetgg of Compound I-1 [DMSO solvate] ] Differential scanning calorimetry of compound I DMSO solvate was measured using the TA Instrument DSC Q2000. A sample (1.77 mg) was weighed in a pinholed aluminum hermetic pan and heated from ambient temperature to 350°C at 10°C/min. The DSC results seen in Figure 3g show a desolvation endotherm at 143°C (onset) followed by a single melting endotherm at 258°C (onset).

Example 12: Compound I-1 [DMAC solvate] Compound I-1° ous form A (100mg; 0.18 mmol), prepared according to the methods described in Example 7, was suspended in DMAC (2000uL) and stirred for 20 hours at 20— °C. The solids were collected by filtration, washed with DMAC (500uL), dried by suction then dried in a vacuum oven at 40—50°C to give compound I-1° DMAC solvate (84mg). 1H NMR (500 MHz, DMSO-d6) 5 10.62 (s, 1H), 9.66 (s, 1H), 9.46 (dd, 1H), 9.26 — 9.22 (m, 1H), 8.24 (d, 1H), 6.77 (s, 2H), 4.54 (t, 2H), 4.46 (t, 2H), 3.66 — 3.54 (m, 4H), 3.43 (p, 1H), 3.18 (tt, 2H), 2.94 (s, 8H), 2.78 (s, 4H), 2.26 (dt, 4H), 2.12 (qd, 2H), 1.96 (s, 4H), 1.76 — 1.69 (m, 2H).

XRPD of Compound I-1 [DMAC e] The XRPD pattern of compound I-1° DMAC e was recorded at room temperature in reflection mode using a ticaZ diffractometer equipped with an an tube source and a PIXcel 1D detector (PANaZyticaZ, The lands). The X-ray generator was ing at a e of 45 kV and a current of 40 mA. The powder sample was placed in a silicon holder. The data were recorded over the range of 3°—3 9° 2 theta with a step size of 0.013° and a dwell time of 121s per step. Figure 1h shows the X-ray powder diffractogram of the sample which is characteristic of crystalline drug substance.

Table 10 depicts representative XRPD peaks form Compound I DMAC solvate: Table 10: Representative XRPD Peaks 2 7.5182 3 8.5957 4 9.7593 10.9655 2014/068713 12.0406 12. 17 —20.85 —71.07 —0.92 —82.12 18.1371 19.0786 19.745 .7384 21.2654 —100 —39.15 —10.68 —15.9 28.763 29.2517 29.5534 29.9062 .5467 31.4852 32.228 44 34.7188 47 38.0592 Thermo Analysis of Compound I-1 gDMAC solvate) A thermograVimetric analysis of compound I-1° DMAC solvate was performed to determine the percent weight loss as a function of temperature using the Discovery TGA (TA Instruments Trios). A sample (5.12mg) was added to a pre—tared aluminum pan and heated from ambient temperature to 350°C at in. The TGA results seen in Figure 2h show a large weight loss of 17.76% between 85°C (onset) and 100°C (end point). This weight loss corresponds to approximately 1.3 molar equivalents of DMAC. The subsequent weight loss seen at 306°C is a result of melting/degradation.

Differential Scanning Calorimetg of Compound I-1 [DMAC solvate] Differential scanning metry of compound I-1° DMAC solvate was measured using the TA Instrument DSC Q2000. A sample (1.93 mg) was d in a pinholed aluminum hermetic pan and heated from ambient temperature to 350°C at 10°C/min. The DSC results seen in Figure 3h show a desolvation endotherm at 81°C ) followed by a single melting endotherm at 261°C (onset).

Example 13: Compound I-1 [acetone e] Compound I-1° amorphous (100mg; 0.18mmol), prepared according to the methods described in e 2, Step 3, above, was suspended in e (2000uL) and stirred for 22 hours.

Compound I-1° acetone solvate was collected by filtration. 1H NMR (500 MHz, DMSO—d6) 5 10.63 (s, 1H), 9.66 (s, 1H), 9.46 (dd, 1H), 9.24 (dd, 1H), 8.24 (d, 1H), 6.78 (s, 2H), 4.54 (t, 2H), 4.46 (t, 2H), 3.65 — 3.54 (m, 4H), 3.43 (p, 1H), 3.19 (tt, 2H), 3.06 — 2.90 (m, 3H), 2.26 (dt, 4H), 2.18 — 2.05 (m, 3H), 1.72 (d, 2H).

XRPD of Compound I-1 [acetone solvate] The XRPD pattern of compound I-1° acetone solvate was recorded at room temperature in reflection mode using a PANalyticaZ diffractometer equipped with an an tube source and a PIXcel 1D detector (PANaZyticaZ, The Netherlands). The X-ray generator was operating at a voltage of 45 kV and a current of 40 mA. The powder sample was placed in a silicon holder. The data were ed over the range of 3°—3 9° 2 theta with a step size of 0.0130 and a dwell time of 121s per step. Figure 1i shows the X-ray powder diffractogram of the sample which is characteristic of crystalline drug substance.

] Table 11 depicts representative XRPD peaks form Compound I-1° acetone solvate: Table 10: Representative XRPD Peaks .4145 7.38 —-1m * 16.6652 —22.77 11 17.1217 —6.15 12 17.9563 —10.57 18.589 19.5447 .0055 .8656 21.3488 22.2722 22 23.465 —3.74 —2.49 —4.74 —13.52 —8.03 —6.04 .6427 31.36 32.2601 33.3871 33.8459 34.2253 .6517 39 35.9083 40 36.4752 Thermo Analysis of Compound I-1 gacetone solvate) A graVimetric analysis of compound I-1° acetone solvate was performed to determine the percent weight loss as a function of temperature using the Discovery TGA (TA Instruments Trios). A sample (2.45mg) was added to a pre-tared aluminum pan and heated from ambient temperature to 350°C at in. The TGA results seen in Figure 2i show an initial weight loss of 1.46%. A larger weight loss of 4.55% was then seen between 124°C (onset) and 151°C (end point), which corresponds to approximately 0.44 molar equivalents of Acetone. The uent weight loss seen at 302°C is a result of g/degradation.

Differential Scanning Calorimetg of nd I-1 [acetone solvate] Differential scanning calorimetry of compound I-1° acetone solvate was measured using the TA Instrument DSC Q2000. A sample (1.42 mg) was weighed in a pinholed um hermetic pan and heated from ambient temperature to 350°C at 10°C/min. The DSC results seen in Figure 3i show a desolvation endotherm at 136 °C (onset) followed by a melting erm at 166°C ).

This was in turn followed by immediate recrystallization exotherm at 175°C. Another melting endotherm was then recorded at 259°C. This was also ed by a recrystallization exotherm at 261°C. A final melting endotherm was observed at 279°C.

Example 14: Compound I-1 [isopropanol solvate] Compound I-1° amorphous (100mg; 0.18mmol), prepared according to the methods described in e 2, Step 3, above, was suspended in anol (2000uL) and stirred for 22 hours at 20—25°C. Compound I-1° isopropanol solvate was collected by tion.

XRPD of Compound I-1 [isopropanol solvate) The XRPD pattern of compound I panol solvate was recorded at room temperature in reflection mode using a PANaZytical diffractometer equipped with an Empyrean tube source and a PIXcel 1D detector (PANaZyticaZ, The Netherlands). The X—ray tor was operating at a voltage of 45 kV and a current of 40 mA. The powder sample was placed in a silicon holder. The data were recorded over the range of 3°—39° 2 theta with a step size of 0.013° and a dwell time of 121s per step. Figure 1j shows the X-ray powder diffractogram of the sample which is characteristic of crystalline drug substance.

Table 12 depicts representative XRPD peaks form Compound I-1° isopropanol solvate: Table 12: Representative XRPD Peaks 6.937 — 11.0107 — 12.8255 13.6694 3.53 14.1876 2.27 .- 14.8878 16.1846 17.1027 18.84 9* 172424 180956 047 272638 28.8751 .0648 31.4229 32.9263 34.4773 .6844 27 37.3825 Thermo Analysis of Compound I-1 opanol e] A thermograVimetric analysis of Compound I-1° isopropanol solvate was performed to ine the percent weight loss as a function of temperature using the Discovery TGA (TA Instruments Trios). A sample (3.39mg) was added to a pre-tared aluminum pan and heated from ambient temperature to 300°C at 10°C/min. The TGA results seen in Figure 2j show a large weight loss of 3.76% between 136°C (onset) and 180°C (end point). This weight loss corresponds to approximately 0.35 molar lents of IPA. The subsequent weight loss seen at 278°C is a result of melting/degradation.

Differential Scanning Calorimetg of Compound I-1 [isopropanol solvate) Differential scanning calorimetry of nd I-1° isopropanol solvate was measured using the TA Instrument DSC Q2000. A sample (1.03 mg) was weighed in a T-zero um pan and heated from ambient temperature to 320°C at 10°C/min. The DSC results seen in Figure 3j show a broad ation endotherm at 135°C (onset) followed by a single melting endotherm at 258°C (onset).

Example 15: Cellular ATR Inhibition Assay: Compounds can be ed for their ability to inhibit intracellular ATR using an immunofluorescence microscopy assay to detect phosphorylation of the ATR substrate histone H2AX in hydroxyurea treated cells. HT29 cells are plated at 14,000 cells per well in 96—well black imaging plates (BD 353219) in McCoy’s 5A media (Sigma M8403) supplemented with 10% foetal bovine serum (JRH Biosciences , Penicillin/Streptomycin solution diluted 1:100 (Sigma P7539), and 2mM L-glumtamine (Sigma G7513), and allowed to adhere ght at 37°C in 5% C02. Compounds are then added to the cell media from a final concentration of 25uM in 3—fold serial dilutions and the cells are incubated at 37°C in 5% C02. After 15min, hydroxyurea (Sigma H8627) is added to a final concentration of 2mM.

After 45min of ent with hydroxyurea, the cells are washed in PBS, fixed for 10min in 4% formaldehyde diluted in PBS (Polysciences Inc 18814), washed in 0.2% Tween—20 in PBS (wash buffer), and permeabilised for 10min in 0.5% Triton X-100 in PBS, all at room temperature.

The cells are then washed once in wash buffer and blocked for 30min at room temperature in 10% goat serum (Sigma G9023) diluted in wash buffer (block buffer). To detect H2AX phosphorylation levels, the cells are then incubated for 1hr. at room temperature in primary antibody (mouse monoclonal hosphorylated e H2AX Ser139 antibody; Upstate 05—636) diluted 1:250 in block buffer. The cells are then washed five times in wash buffer before incubation for 1hr. at room temperature in the dark in a mixture of secondary antibody (goat ouse Alexa Fluor 488 conjugated antibody; Invitrogen A11029) and Hoechst stain (Invitrogen H3570); diluted 1:500 and 1:5000, respectively, in wash . The cells are then washed five times in wash buffer and finally 100ul PBS is added to each well before imaging.

Cells are imaged for Alexa Fluor 488 and Hoechst intensity using the BD Pathway 855 Bioimager and Attovision software (BD Biosciences, Version 1.6/855) to quantify phosphorylated H2AX Ser139 and DNA staining, respectively. The percentage of phosphorylated H2AX-positive nuclei in a montage of 9 images at 20x magnification is then calculated for each well using BD Image Data er software (BD Biosciences Version 2.2.15). Phosphorylated H2AX—positive nuclei are defined as t-positive s of interest containing Alexa Fluor 488 intensity at 1.75-fold the average Alexa Fluor 488 intensity in cells not treated with hydroxyurea. The percentage of H2AX positive nuclei is finally plotted against concentration for each compound and IC50s for intracellular ATR inhibition are determined using Prism software (GraphPad Prism version 3.0cx for Macintosh, GraphPad re, San Diego California, USA).

The compounds described herein can also be tested according to other methods known in the art (@ Sarkaria et al, “Inhibition ofATM and ATR Kinase Activities by the Radiosensitizing Agent, Caffeine: Cancer Research 59: 3 82 (1999); n et al, “Identification and Characterization of a Novel and Specific Inhibitor of the Ataxia-Telangiectasia Mutated Kinase ATM” Cancer Research 64: 159 (2004); Kim et al, “Substrate Specificities and Identification of ve Substrates ofATM Kinase Family Members” The Journal ofBiological Chemistry, 274(53): 3753 8—3 7543 (1999); and Chiang et al, “Determination of the catalytic activities of mTOR and other members of the phosphoinositide-3 -kinase—related kinase family” Methods Mol. Biol. 281 : 125—41 (2004)).

Example 16: ATR Inhibition Assay: Compounds can be screened for their ability to inhibit ATR kinase using a radioactive— phosphate incorporation assay. Assays are carried out in a mixture of 50mM Tris/HCl (pH 7.5), 10mM MgClz and 1mM DTT. Final substrate trations are 10uM [y-33P]ATP (3mCi 33P ATP/mmol ATP, Perkin Elmer) and 800 [1M target peptide (ASELPASQPQPFSAKKK).

Assays are carried out at 25°C in the presence of 5 nM full-length ATR. An assay stock buffer solution is prepared containing all of the reagents listed above, with the exception of ATP and the test compound of interest. 13.5 [1L of the stock solution is placed in a 96 well plate followed by addition of 2 [1L of DMSO stock containing serial dilutions of the test compound (typically starting from a final concentration of 15 [1M with 3-fold serial dilutions) in duplicate (final DMSO concentration 7%). The plate is cubated for 10 minutes at 25°C and the reaction initiated by on of 15 [LL [y-33P]ATP (final concentration 10 uM).

The reaction is stopped after 24 hours by the addition of 30uL 0.1M phosphoric acid containing 2mM ATP. A multiscreen phosphocellulose filter l plate (Millipore, Cat no.

MAPHNOB50) is pretreated with 100uL 0.2M phosphoric acid prior to the addition of 45 [LL of the stopped assay mixture. The plate is washed with 5 x 200uL 0.2M phosphoric acid. After drying, 100 [1L Optiphase ‘SuperMix’ liquid scintillation cocktail n Elmer) is added to the well prior to scintillation counting (1450 Microbeta Liquid Scintillation Counter, Wallac).

After removing mean background values for all of the data points, Ki(app) data are calculated from near regression is of the initial rate data using the Prism software package (GraphPad Prism version 3.0cx for Macintosh, GraphPad re, San Diego California, USA).

In general, the compounds of the present invention are effective for inhibiting ATR.

Compounds I-1 and L3 inhibit ATR at Ki values below 1 uM.

Example 17: Cisplatin Sensitization Assay nds can be ed for their ability to sensitize HCT116 ctal cancer cells to Cisplatin using a 96h cell viability (MTS) assay. HCT116 cells, which possess a defect in ATM signaling to Cisplatin (see, Kim et al.; Oncogene 21:3 864 (2002); see also, Takemura et al.; JBC 281 :30814 (2006)) are plated at 470 cells per well in 96—well polystyrene plates (Costar 3596) in 150ul of McCoy’s 5A media (Sigma M8403) supplemented with 10% foetal bovine serum (JRH Biosciences 12003), Penicillin/Streptomycin solution diluted 1:100 (Sigma P753 9), and 2mM L- glumtamine (Sigma G7513), and allowed to adhere overnight at 37°C in 5% C02. Compounds and Cisplatin are then both added simultaneously to the cell media in 2-fold serial dilutions from a top final concentration of 10uM as a full matrix of concentrations in a final cell volume of 200ul, and the cells are then incubated at 37°C in 5% C02. After 96h, 40ul of MTS reagent (Promega G358a) is added to each well and the cells are incubated for 1hr. at 37°C in 5% C02. Finally, absorbance is measured at 490nm using a SpectraMax Plus 384 reader ular Devices) and the tration of compound required to reduce the IC50 of Cisplatin alone by at least 3—fold (to 1 decimal place) can be reported.

In general, the compounds of the present invention are effective for sensitizing cancer cells to Cisplatin. Compounds I-1 and L3 have Cisplatin sensitization values of < 0.2 uM.

Example 18: Single Agent HCT116 Activity Compounds can be screened for single agent activity against HCT116 colorectal cancer cells using a 96h cell viability (MTS) assay. HCT116 are plated at 470 cells per well in 96-well polystyrene plates (Costar 3596) in 150ul of McCoy’s 5A media (Sigma M8403) supplemented with % foetal bovine serum (JRH Biosciences 12003), Penicillin/ omycin on diluted 1:100 (Sigma P7539), and 2mM L-glumtamine (Sigma G7513), and allowed to adhere ght at 37°C in % C02. nds are then added to the cell media in 2—fold serial dilutions from a top final concentration of 10uM as a full matrix of concentrations in a final cell volume of 200ul, and the cells are then incubated at 37°C in 5% C02. After 96h, 40ul of MTS reagent (Promega G358a) is added to each well and the cells are incubated for 1hr. at 37°C in 5% C02. y, absorbance is measured at 490nm using a SpectraMax Plus 384 reader (Molecular s) and IC50 values can be ated.

Example 19: ATR-complex Inhibition Assay Compounds were screened for their ability to inhibit ATR kinase, in the presence of r proteins ATRIP, CLK2 and TopBPl, using a radioactive-phosphate incorporation assay. Assays were carried out in a mixture of 50 mM Tris/HCl (pH 7.5), 10 mM MgClz and 1 mM DTT. Final substrate trations were 10 ”M [g-33P]ATP (3.5 ”Ci 33P ATP/nmol ATP, Perkin Elmer, Massachusetts, USA) and 800 ”M target peptide (ASELPASQPQPFSAKKK, Isca Biochemicals, Cambridgeshire, UK).

Assays were carried out at 25°C in the presence of 4 nM full—length ATR, 40 nM full- length ATRIP, 40 nM full-length CLK2 and 600 nM TopBP1(A891-S1105). An enzyme stock buffer solution was prepared ning all of the reagents listed above, with the exception of target peptide, ATP and the test compound of st. This enzyme stock was pre—incubated for 30 minutes at 25°C. 8.5 uL of the enzyme stock solution was placed in a 96—well plate followed by addition of 5ul of target peptide and 2 uL of DMSO stock containing serial dilutions of the test compound (typically starting from a final concentration of 1.5 ”M with 2.5-fold serial dilutions) in duplicate (final DMSO concentration 7%). The plate was pre-incubated for 10 minutes at 25°C and the reaction initiated by addition of 15 uL [g-33P]ATP (final concentration 10 uM).

The reaction was d after 20 hours by the addition of 30 uL 0.3 M phosphoric acid containing 2 mM ATP. A phosphocellulose filter 96-well plate (Multiscreen HTS MAPHNOB50, Merck-Millipore, Massachusetts, USA) was pretreated with 100 uL 0.1 M phosphoric acid prior to the on of 45 uL of the stopped assay mixture. The plate was washed with 5 x 200 ”L 0.1 M phosphoric acid. After drying, 50 uL Optiphase ‘SuperMix’ liquid scintillation il (Perkin Elmer, Massachusetts, USA) was added to the well prior to scintillation counting (Wallac 1450 Microbeta Liquid Scintillation Counter, Perkin Elmer, Massachusetts, USA).

After removing mean background values for all of the data points, ) data were calculated from near sion analysis of the initial rate data using the Prism software e (GraphPad Prism version 6.0c for Macintosh, GraphPad Software Inc., San Diego, USA).

While we have described a number of embodiments of this invention, it is apparent that our basic examples may be d to provide other embodiments that utilize the compounds, methods, and processes of this invention. ore, it will be appreciated that the scope of this invention is to be defined by the appended claims rather than by the specific embodiments that have been represented by way of example herein.

Claims (68)

CLAIMS 1. We claim:

1. A solid form of a compound of formula I-1: wherein the form is selected from the group consisting of Compound I-1 hydrate I, Compound I-1 anhydrous form A, Compound I-1 anhydrous form B, Compound I-1 anhydrous form C, Compound I-1 DMSO solvate, Compound I-1 DMAC solvate, Compound I-1 acetone solvate, or Compound I-1 isopropanol solvate, wherein nd I-1 hydrate I is crystalline Compound I-1 hydrate I terized by one or more peaks sed in 2-theta ± 0.2 at 6.5, 12.5, 13.7, 18.8, and 26.0 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation, Compound I-1 anhydrous form A is crystalline Compound I-1 ous form A characterized by one or more peaks sed in 2-theta ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation, nd I-1 anhydrous form B is crystalline Compound I-1 anhydrous form B characterized by two or more peaks expressed in 2-theta ± 0.2 at 7.2, 8.3, 12.9, 19.5, and 26.6 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation, nd I-1• anhydrous form C is lline Compound I-1• anhydrous form C characterized by two or more peaks expressed in 2-theta ± 0.2 at 6.8, 13.4, 15.9, 30.9, and 32.9 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation.

2. The solid form of claim 1, wherein the form is crystalline Compound I-1• hydrate I.

3. The solid form of claim 2, wherein the crystalline Compound I-1• hydrate I has a Compound I-1 to H2O ratio of about 1:4.5.

4. The solid form of claim 2, characterized by a weight loss of from about 14.56% in a temperature range of from about 25 °C to about 100 °C as determined by a thermal etric analysis.

5. The solid form of claim 2, characterized as having the following peaks in a X-ray powder diffraction pattern Angle Intensity % (2-Theta ± 0.2) 4.0 1.4 4.8 3.0 5.7 7.8 6.3 100.0 6.5 51.0 9.1 3.0 10.1 10.5 10.4 10.8 11.2 5.9 11.5 8.7 11.8 11.5 12.5 16.0 13.7 10.9 14.3 7.2 15.0 9.2 15.5 10.5 16.9 14.8 18.8 10.8 20.1 14.1 20.6 11.6 22.6 10.2 23.9 4.5 24.7 7.8 26.0 13.6 27.3 10.9 28.6 4.9 32.3 2.2.

6. The solid form of claim 1, wherein the form is crystalline Compound I-1• anhydrous form A.

7. The solid form of claim 6, characterized by a weight loss of from about 0.96 % in a temperature range of from about 25 °C to about 265 °C as determined by a thermal etric analysis.

8. The solid form of claim 6, characterized as having the following peaks in a X-ray powder diffraction pattern Angle Intensity % (2-theta ± 0.2) 3.6 12.5 3.9 17.4 6.1 51.0 9.7 20.5 12.2 22.8 14.0 23.5 14.5 22.2 16.4 33.5 17.1 25.0 17.8 36.0 19.1 21.5 20.2 26.5 21.3 16.1 22.3 31.6 24.4 23.7 25.3 100.0 28.4 11.9 31.8 16.0.

9. The solid form of claim 6, characterized as having one or more peaks corresponding to 175.9 ± 0.3 ppm, 138.9 ± 0.3 ppm, 74.1 ± 0.3 ppm, 42.8 ± 0.3 ppm, and 31.5± 0.3 ppm in a 13C ssNMR spectrum.

10. The solid form of claim 6, terized as having one or more peaks corresponding to -136.8 ± 0.3 ppm and -155.7 ± 0.3 ppm in an 19F ssNMR spectrum.

11. The solid form of claim 1, wherein the form is crystalline Compound I-1• anhydrous form B.

12. The solid form of claim 11, characterized by a weight loss of from about 2.5 % in a temperature range of from about 25 °C to about 175 °C as determined by a thermal gravimetric

13. The solid form of claim 11, characterized as having the following peaks in a X-ray powder diffraction pattern Angle (2-theta ± 0.2) Intensity % 5.2 19.0 5.9 33.8 7.2 52.9 8.3 79.0 9.8 88.8 11.1 60.8 11.7 65.4 12.9 62.9 14.8 62.0 15.6 100.0 16.3 62.7 16.8 57.1 18.0 52.6 19.5 33.9 20.4 11.5 21.3 8.3 23.2 22.1 25.2 39.9 25.9 27.3 26.6 22.9 27.4 30.0 28.0 44.8 28.9 26.9 30.6 18.0 32.2 11.6 36.0 3.1.

14. The solid form of claim 11, characterized as having one or more peaks corresponding to 173.4 ± 0.3 ppm, 164.5 ± 0.3 ppm, 133.5 ± 0.3 ppm, 130.8 ± 0.3 ppm, 67.7 ± 0.3 ppm, 45.3 ± 0.3 ppm, and 25.9 ± 0.3 ppm in a 13C ssNMR spectrum.

15. The solid form of claim 11, characterized as having one or more peaks corresponding to - 138.0 ± 0.3 ppm and -153.5 ± 0.3 ppm in an 19F ssNMR spectrum.

16. The solid form of claim 1, wherein the form is crystalline Compound I-1• anhydrous form C.

17. The solid form of claim 16, terized as having the following peaks in a X-ray powder diffraction pattern Angle (2-theta ± 0.2) Intensity % 3.6 1.1 4.0 1.1 6.8 44.8 7.6 11.1 8.1 12.7 10.3 13.3 11.4 100.0 13.4 29.1 14.2 51.8 14.9 23.8 15.9 31.1 16.3 14.2 16.7 17.6 17.0 26.9 18.2 37.9 19.1 50.4 20.7 31.7 22.6 3.8 23.3 16.0 23.9 15.0 24.5 10.1 25.5 24.0 25.8 33.3 27.1 17.3 27.9 23.8 29.1 19.2 30.9 22.3 32.0 12.9 32.9 12.8 33.7 7.5 35.1 4.5.

18. The solid form of claim 16, characterized as having one or more peaks corresponding to 175.2 ± 0.3 ppm, 142.5 ± 0.3 ppm, 129.6 ± 0.3 ppm, 73.5± 0.3 ppm, 54.0 ± 0.3 ppm, and 46.7 ± 0.3 ppm in a 13C ssNMR spectrum.

19. The solid form of claim 16, characterized as having one or more peaks corresponding to - 131.2 ± 0.3 ppm and -150.7 ± 0.3 ppm in an 19F ssNMR spectrum.

20. The solid form of claim 1, wherein the form is Compound I-1• DMSO solvate.

21. The solid form of claim 1, wherein the form is lline Compound I-1• DMSO solvate.

22. The solid form of claim 21, wherein the crystalline Compound I-1• DMSO solvate has a compound I-1 to DMSO ratio of about 1:1.

23. The solid form of claim 21, characterized by a weight loss of from about 12.44% in a temperature range of from about 146°C to about 156°C as determined by a thermal gravimetric analysis.

24. The solid form of claim 21, characterized by one or more peaks expressed in 2-theta ± 0.2 at 8.9, 14.8, 16.5, 18.6, 20.9, 22.2, and 23.4 degrees in an X-Ray powder ction pattern ed using Cu K alpha radiation.

25. The solid form of claim 21, characterized as having the ing peaks in a X-ray powder diffraction pattern Angle (2-theta ± 0.2) Intensity % 7.0034 8.33 8.9204 11.28 10.4007 10.11 12.4735 6.81 12.7962 12.32 13.3976 12.25 14.8102 29.16 15.439 15.1 15.7477 14.37 16.5454 82.57 17.051 15.34 18.1033 20.25 18.6354 29.71 19.593 3.68 20.1178 6.42 20.9143 35.76 21.3593 11.65 22.1801 100 22.8306 25.4 23.3866 51.08 23.8312 16.31 24.5088 15.65 25.6545 25.59 27.0136 3.06 27.4405 2.43 27.6871 3.27 5 8.73 28.9693 11.53 29.555 8.95 30.1186 5.69 30.5402 8.63 31.2969 6.42 32.0663 8.71 33.2165 3.04 34.1902 7.02 34.6067 3.57 35.45 1.47 36.5669 3.23 2 2.19.

26. The solid form of claim 1, wherein the form is Compound I-1• DMAC solvate.

27. The solid form of claim 1, wherein the form is lline Compound I-1• DMAC solvate.

28. The solid form of claim 27, wherein the crystalline Compound I-1• DMAC solvate has a compound I-1 to DMAC ratio of about 1:1.3.

29. The solid form of claim 27, characterized by a weight loss of from about 17.76% in a temperature range of from about 85°C to about 100°C as determined by a thermal gravimetric analysis.

30. The solid form of claim 27, characterized by one or more peaks sed in 2-theta ± 0.2 at 6.0, 15.5, 17.7, 18.1, 20.4, and 26.6 degrees in an X-Ray powder diffraction pattern ed using Cu K alpha radiation.

31. The solid form of claim 27, characterized as having the ing peaks in a X-ray powder diffraction pattern Angle (2-theta ± 0.2) Intensity % 6.0169 75.51 7.5182 7.99 8.5957 32.29 9.7593 33.98 10.9655 15.95 11.3688 7.25 12.0406 12.17 13.6703 19.18 14.1108 36.56 14.2831 23.2 14.5895 9.33 15.1755 25.52 15.4632 20.85 16.0919 71.07 16.9423 0.92 17.7117 82.12 18.1371 77.28 18.5857 4.73 19.0786 16.95 19.745 7.05 20.3531 40.38 20.7384 29.95 21.2654 10.22 21.7978 9.56 22.4818 2.27 22.8051 5.51 23.3945 6.33 23.829 19.65 24.6486 3.69 25.343 5.43 2 7.83 26.6041 100 27.6488 39.15 28.1311 10.68 28.4779 15.9 28.763 13.68 29.2517 17.62 29.5534 13.91 29.9062 12.28 30.5467 7.27 31.4852 9.17 32.228 2.69 32.6692 3.7 34.7188 1.29 36.4642 1.43 37.1111 1.9 38.0592 1.92.

32. The solid form of claim 1, wherein the form is Compound I-1• acetone solvate.

33. The solid form of claim 1, wherein the form is crystalline Compound I-1• acetone solvate.

34. The solid form of claim 33, wherein the crystalline Compound I-1• e solvate has a compound I-1 to acetone ratio of about 1:0.44.

35. The solid form of claim 33, characterized by a weight loss of from about 4.55% in a ature range of from about 124°C to about 151°C as determined by a thermal etric analysis.

36. The solid form of claim 33, characterized by one or more peaks expressed in 2-theta ± 0.2 at 8.9, 15.5, 15.8, 16.7, 22.3, 25.7, and 29.0 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation.

37. The solid form of claim 33, characterized as having the following peaks in a X-ray powder ction pattern Angle (2-theta ± 0.2) Intensity % 6.9871 31.75 8.9148 62.84 10.4145 7.38 12.4529 6.65 12.7486 9.09 13.4567 7.37 14.8093 10.97 15.528 35.3 15.826 19.22 16.6652 22.77 17.1217 6.15 17.9563 10.57 18.1349 9.4 18.589 7.22 19.5447 3.06 20.0055 2.55 6 6.29 21.3488 6.36 22.2722 100 5 13.43 22.9581 19.8 23.465 21.26 23.7918 8.65 24.5843 8.65 25.7222 13.01 26.0003 3.74 27.696 2.49 28.7335 4.74 29.0658 13.52 29.6743 8.03 30.2154 6.04 30.6427 4.67 31.36 4.28 32.2601 3.86 33.3871 0.66 33.8459 1.15 34.2253 1.42 35.6517 2.34 35.9083 2 36.4752 2.17.

38. The solid form of claim 1, wherein the form is Compound I-1• isopropanol solvate.

39. The solid form of claim 1, wherein the form is crystalline Compound I-1• panol solvate.

40. The solid form of claim 39, wherein the crystalline nd I-1• isopropanol solvate has a compound I-1 to isopropanol ratio of about .

41. The solid form of claim 39, characterized by a weight loss of from about 3.76% in a temperature range of from about 136°C to about 180°C as determined by a thermal gravimetric analysis.

42. The solid form of claim 39, characterized by one or more peaks expressed in 2-theta ± 0.2 at 6.9, 17.1, 17.2, 19.1, 19.6, 23.7, 24.4, and 28.9 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation.

43. The solid form of claim 39, characterized as having the ing peaks in a X-ray powder diffraction pattern Angle (2-theta ± 0.2) Intensity % 6.937 100 7 7.85 12.8255 8.34 13.6694 3.53 14.1876 2.27 14.8878 7.9 16.1846 4.17 17.1027 18.84 17.2424 19.04 6 0.47 19.1139 5.27 19.6437 15.33 20.3628 10.96 21.4978 1.13 22.769 5.81 23.6531 41.5 24.3573 39.72 24.8556 17.48 25.8121 8.63 27.2638 2.35 28.8751 21.82 30.0648 2.34 31.4229 1.58 32.9263 1.14 34.4773 2.29 35.6844 1.53 37.3825 0.46.

44. A composition sing: a) Compound I-1, or a pharmaceutically acceptable salt thereof, wherein Compound I-1 is represented by the following structural formula: ; and b) one or more excipients, wherein at least 90% by weight of Compound I-1 is crystalline Compound I-1•anhydrous form A characterized by one or more peaks expressed in 2-theta ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation.

45. The composition of claim 44, wherein the one or more excipients comprises one or more selected from the group consisting of one or more s, one or more g agents, one or more lubricants, and one or more disintegrants.

46. The ition of claims 44 or 45, wherein the one or more excipients comprise one or more fillers.

47. The composition of claim 46, wherein the one or more fillers is present in an amount in the range of about 10 wt% to about 88 wt% by the total weight of the composition.

48. The composition of claim 46 or 47, wherein the one or more fillers is selected from the group consisting of mannitol, lactose, sucrose, dextrose, maltodextrin, sorbitol, xylitol, powdered cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, methylhydroxyethylcellulose, starch, pregelatinized , dibasic calcium phosphate, calcium sulfate and m ate.

49. The composition of claim 48, wherein the one or more filler is selected from microcrystalline cellulose and lactose.

50. The ition of any one of claims 44-49, wherein the one or more excipients comprises one or more disintegrants.

51. The composition of claim 50, wherein one or more disintegrants is present in an amount in the range of about 1 wt% to about 15 wt% by the total weight of the composition.

52. The composition of claim 50 or 51, wherein the one or more disintegrants is selected from the group consisting of croscarmellose sodium, sodium alginate, calcium alginate, alginic acid, starch, pregelatinized starch, sodium starch glycolate, crospovidone, cellulose and its tives, carboxymethylcellulose calcium, carboxymethylcellulose sodium, soy ccharide, guar gum, an ion exchange resin, an effervescent system based on food acids and an alkaline carbonate component, and sodium bicarbonate.

53. The composition of claim 52, wherein the one or more disintegrants is croscarmellose

54. The composition of any one of claims 44-53, wherein the one or more excipients comprises one or more lubricants.

55. The composition of claim 54, wherein the one or more lubricants is present in an amount in the range of about 0.1 wt% to about 10 wt% by the total weight of the composition.

56. The composition of claim 54 or 55, wherein the one or more lubricants is ed from the group consisting of talc, fatty acid, stearic acid, magnesium stearate, calcium stearate, sodium stearate, glyceryl monostearate, sodium lauryl sulfate, sodium l fumarate, hydrogenated oils, fatty alcohol, fatty acid ester, glyceryl behenate, mineral oil, vegetable oil, leucine, sodium benzoate, and a combination f.

57. The ition of claim 56, wherein the one or more lubricants is sodium stearyl fumarate.

58. The ition of any one of claims 44-57, comprising: a) an amount of Compound I-1 in the range of about 5 wt% to about 50 wt% by the total weight of the composition; b) an amount of one or more lubricants in the range of about 0.1 wt% to about 10 wt% by the total weight of the composition; c) an amount of one or more disintegrants in the range of about 1 wt% to about 15 wt% by the total weight of the composition; and d) an amount of one or more fillers in the range of about 10 wt% to about 90 wt% by the total weight of the composition.

59. The composition of any one of claims 44-58, comprising: a) an amount of Compound I-1 of 10.0 wt% by the total weight of the composition; b) an amount of lactose monohydrate of 27.5 wt% by the total weight of the composition; c) an amount of Avicel PH-101 (microcrystalline cellulose) of 55.0 wt% by the total weight of the composition; d) an amount of Sol (croscarmellose sodium) of 4.5 wt% by the total weight of the composition; and e) an amount of sodium stearyl fumarate of 3.0 wt% by the total weight of the composition.

60. The composition of any one of claims 44-59, wherein all of Compound I-1 is Form A.

61. The composition of any one of claims 44-59, wherein at least 95% by weight of Compound I- 1 is Form A.

62. The composition of claim 61, wherein at least 98% by weight of Compound I-1 is Form A.

63. A l form of Compound I-1 having a monoclinic crystal system, a P21/c centrosymmetric space group, and the following unit cell ters: a = 3)Å α = 90° b = 12.17(2)Å β = 107.22(3)° c = 14.48(3)Å γ = 90° wherein Compound I-1 is represented by the following structural formula:

64. A process for preparing Compound I-1•anhydrous form A comprising stirring a suspension containing Compound I-1•ethanol solvate and tetrahydrofuran wherein Compound I-1•anhydrous form A is crystalline Compound I-1•anhydrous form A characterized by one or more peaks expressed in a ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X-Ray powder ction pattern obtained using Cu K alpha radiation, n Compound I-1 is represented by the following structural formula:

65. A process for preparing Compound I-1•anhydrous form A comprising stirring a suspension containing nd I-1•amorphous, isopropanol, and water, wherein Compound I-1•anhydrous form A is crystalline nd I-1•anhydrous form A characterized by one or more peaks expressed in 2-theta ± 0.2 at 6.1, 12.2, 14.5, 22.3, and 31.8 degrees in an X-Ray powder diffraction pattern obtained using Cu K alpha radiation, n Compound I-1 is represented by the following structural formula:

66. The process of claim 65, wherein the suspension is heated to between about 65°C and about 80°C.

67. The process of claim 66, wherein the suspension is heated to between about 70°C and about 75°C.

68. A solid form of claim 1, substantially as herein described with reference to any one of the es and/or