Crystalline and amorphous forms of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl)-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and its salts, and methods for their preparation and therapeutic uses

Abstract

Crystalline and amorphous forms of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine (Compound 1) and various salts thereof, methods for preparing these crystalline and amorphous forms, pharmaceutical compositions containing these crystalline and amorphous forms, and uses of these crystalline and amorphous forms or pharmaceutical compositions thereof for the treatment of diseases or disorders associated with cyclin-dependent kinase (CDK), particularly CDK4 and CDK6, activity, such as various cancers.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application No. 62 / 821,141, filed March 20, 2019, the disclosure of which is incorporated herein by reference in its entirety. Technical Field The present invention relates to crystalline and amorphous forms of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl)-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and pharma- ceutically acceptable salts thereof, and their use in the treatment or prevention of diseases or conditions mediated by cyclin-dependent kinases (CDKs), such as various cancers. [Background technology]

[0002] Cyclin-dependent kinases (CDKs) are a family of protein kinases that regulate cell division and proliferation. Cell cycle progression is controlled by cyclins and their associated cyclin-dependent kinases, such as CDK1-CDK4 and CDK6, whereas other CDKs, such as CDK7-CDK9, are important for transcription. CDKs that bind to cyclins form heterodimeric complexes that phosphorylate substrates on serine and threonine residues, thereby initiating events required for transcription and progression through the cell cycle (Malumbres et al., Trends Biochem. Sci. 2005, 30, 630-641). Because uncontrolled cell proliferation is a hallmark of cancer and most cancer cells exhibit deregulation of CDKs, inhibition of CDKs has emerged as a potential treatment for various cancers. Although inhibitors with different degrees of selectivity for CDKs have been reported, selective CDK4 / 6 inhibitors are currently considered a promising class of potential cancer therapeutics due to the important role of CDK4 / 6 in regulating cell proliferation and the toxic effects associated with inhibition of other CDKs. Compound 1, N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine, is a potent selective inhibitor of CDK4 / 6 useful for treating or preventing diseases, disorders, or conditions mediated by certain CDKs, particularly CDK4 and CDK6, such as various types of cancer and inflammation-related conditions. See WO2016 / 014904 A1 and U.S. Pat. No. 9,878,994, which are incorporated by reference herein in their entireties. [ka] Summary of the Invention [Problem to be solved by the invention]

[0003] However, there remains a need to identify suitable solid forms that can exhibit desirable chemical and physical properties to develop Compound 1 into a viable therapeutic agent. It will also be essential to develop reliable and reproducible methods for producing and purifying Compound 1 in order to develop suitable formulations. However, finding the appropriate salt and / or crystalline form to produce a formulation with suitable physical and biological properties, including but not limited to solubility, stability, melting point, bioavailability, etc., is often not completely predictable. The present invention is directed to meeting such a need. [Means for solving the problem]

[0004] Summary of the invention The present invention provides various solid forms of the compound of formula 1 (Compound 1), including amorphous and crystalline forms, in free base or salt form, and methods for making them. This application discloses physical form data for the characterization of Compound 1 and its various pharmaceutical salt forms and polymorphs.

[0005] In one embodiment, the present invention provides a solid form of Compound 1 as the free base: [ka]

[0006] The free base of Compound 1 exists in one or more crystalline forms, designated as crystalline Form A, Form B, Form C, Form D, Form E, and Form F, respectively.

[0007] In another aspect, the present invention provides a pharma- ceutically acceptable salt of Compound 1 comprising Compound 1 and an acid, including, but not limited to, HCl, H2SO4, H3PO4, methanesulfonic acid, and p-toluenesulfonic acid. Any of the salts may exist amorphous or in various crystalline forms.

[0008] In one embodiment, the present invention provides the hydrochloride salt of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine, wherein compound 1 and the hydrochloride salt are in a molar ratio of about 1:1.

[0009] In another aspect, the invention provides the sulfate salt of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0010] In one embodiment, the sulfate salt comprises N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and sulfuric acid in a molar ratio of about 1:1.

[0011] In another embodiment, the sulfate salt comprises N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and sulfuric acid in a molar ratio of about 2:1.

[0012] In some embodiments, the sulfate salt is present in a solid amorphous form in a molar ratio of about 1:1 or about 2:1.

[0013] In some embodiments, the sulfate salt is present in various crystalline forms (polymorphs) in a molar ratio of about 1:1 or about 2:1.

[0014] In another aspect, the present invention provides the phosphate salt of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0015] In one embodiment, the phosphate salt comprises N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and phosphoric acid in a molar ratio of about 1:1.

[0016] In another aspect, the present invention provides the toluenesulfonic acid (tosylate) salt of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0017] In one embodiment, the tosylate salt comprises -toluenesulfonic acid and N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and phosphoric acid in a molar ratio of about 1:1.

[0018] In another aspect, the present invention provides the methanesulfonic acid (mesylate) salt of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0019] In one embodiment, the mesylate salt comprises N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and methanesulfonic acid in a molar ratio of about 1:2.

[0020] In another embodiment, the mesylate salt comprises N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and methanesulfonic acid in a molar ratio of about 1:1.

[0021] In some embodiments, the mesylate salt is present in a solid amorphous form in a molar ratio of about 1:1 or about 2:1.

[0022] In some embodiments, the mesylate salt exists in various crystalline forms (polymorphs) in a molar ratio of about 1:1 or about 2:1.

[0023] In another aspect, the present invention provides methods for preparing solid amorphous forms of Compound 1 and its salts, particularly the HCl, H2SO4, H3PO4, methanesulfonic acid, and p-toluenesulfonic acid salts disclosed herein.

[0024] In another aspect, the present invention provides methods for preparing the crystalline forms of Compound 1 and its salts, particularly the HCl, H2SO4, H3PO4, methanesulfonic acid, and p-toluenesulfonic acid salts disclosed herein.

[0025] In another aspect, the present invention provides a pharmaceutical composition comprising an amorphous form of Compound 1, or any of its salts, and a pharma- ceutically acceptable carrier.

[0026] In another aspect, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1, or any of its salts, and a pharma- ceutically acceptable carrier.

[0027] In another aspect, the present invention provides a method of inhibiting a cyclin-dependent kinase (CDK) in a biological sample or subject by using an amorphous or crystalline form of Compound 1, or a salt thereof.

[0028] In another aspect, the present invention provides a method for treating or preventing a disease or disorder in a subject associated with CDK activity, comprising administering to the subject a pharmaceutical composition comprising an amorphous or crystalline form of Compound 1 and a pharma- ceutical acceptable carrier.

[0029] In another aspect, the present invention provides a method for treating or preventing a disease or disorder in a subject associated with CDK activity, comprising administering to the subject a pharmaceutical composition comprising a pharmaceutical salt of Compound 1, in amorphous or crystalline form, and a pharma- ceutical acceptable carrier.

[0030] In any of the above aspects or embodiments, the pharmaceutical salt is preferably selected from the group consisting of HCl, H2SO4, H3PO4, methanesulfonic acid, and p-toluenesulfonic acid salts.

[0031] In some preferred embodiments, the crystalline form of a pharma- ceutically acceptable salt of Compound 1 includes, but is not limited to, crystalline form A, crystalline form B, crystalline form C, crystalline form D, or crystalline form E, as applicable, of the compound of Formula 1 or a salt thereof.

[0032] Diseases or disorders associated with CDK that can be treated according to the present invention include, but are not limited to, brain tumors, metastatic brain tumors, squamous cell carcinoma, bladder cancer, gastric cancer, pancreatic cancer, liver cancer, glioblastoma multiforme breast cancer, head cancer, neck cancer, esophageal cancer, prostate cancer, colorectal cancer, lung cancer, renal cancer, kidney cancer, ovarian cancer, gynecological cancer, thyroid cancer, non-small cell lung cancer (NSCLC), refractory ovarian cancer, or head and neck cancer. In one embodiment, the cyclin-dependent kinase (CDK) is CDK4 or CDK6.

[0033] Other aspects and advantages of the present invention will be better understood in view of the following detailed description, drawings, and claims. [Brief description of the drawings]

[0034] [Figure 1] FIG. 1 shows a PLM image of Compound 1 free base (Form A) raw material. [Diagram 2] 2A and 2B show the X-ray powder diffraction pattern (A); and (B) the peak printout of Compound 1 free base (Form A). [Diagram 3] FIG. 3 shows the DSC and TGA profiles of the free base polymorph of Compound 1 (Form A). [Figure 4] FIG. 4 shows the X-ray powder diffraction patterns of the free base polymorphs of Compound 1 (Forms AD). [Diagram 5] Figures 5A, 5B, 5C, 5D, 5F and 5G show peak listings corresponding to the X-ray powder diffraction patterns of the free base polymorphs of the compound of Formula 1 shown in Figure 4. Panels A-B correspond to Compound 1, Form C, and Panels C-G correspond to Compound 1, Form B. [Figure 6] FIG. 6 shows the DSC and TGA profiles of Compound 1 free base (Form B). [Figure 7] FIG. 7 shows the DSC and TGA profiles of Compound 1 free base (Form C). [Figure 8] FIG. 8 shows the DSC and TGA profiles of Compound 1 free base (Form D). [Figure 9] FIG. 9 shows the dynamic vapor sorption (DVS) of Compound 1 free base (Form D). [Figure 10] FIG. 10 shows the X-ray powder diffraction patterns of Compound 1 free base (Form D) before and after DVS. [Figure 11] FIG. 11 shows the X-ray powder diffraction pattern (wet sample) of a 50 mg salt screen of Compound 1 (free base:acid). [Figure 12] FIG. 12 shows the X-ray powder diffraction pattern of a 50 mg scale salt screen of Compound 1 (free base:acid) (dry sample). [Figure 13] 13A and 13B show (A) the X-ray powder diffraction pattern (dry sample) and (B) the peak printout of 50 mg scale compound 1·H2SO4 (1:1; Form A). [Figure 14] FIG. 14 shows the X-ray powder diffraction pattern (dry sample) of a 50 mg scale salt screen (free base:acid) of Compound 1 (Form A). [Figure 15]15A, 15B, 15C, and 15D show peak printouts of 500 mg scale salt screens (free base:acid) of compound 1 (dry sample) shown in FIG. 14: (A) H2SO4 (2:1; form A) salt, (B) H3PO4 (1:1; form A) salt, (C) HCl (1:1; form A) salt, (D) p-toluenesulfonic acid (1:1; form A) salt. [Figure 16] FIG. 16 shows the TGA and DSC profiles of compound 1·HCl (1:1; Form A) (dry sample). [Figure 17] FIG. 17 shows the DVS isotherm analysis of compound 1·HCl (1:1; Form A). [Figure 18] 18A, 18B, and 18C show the X-ray powder diffraction patterns of compound 1·HCl (1:1; Form A) before and after DVS. (B) Peak printout before DVS. (C) Peak printout after DVS. [Figure 19] FIG. 19 shows the 400 MHz 1H NMR of compound 1·HCl (1:1; Form A) in DMSO-d6. [Figure 20] FIG. 20 shows the TGA and DSC profiles of compound 1·H2SO4 (1:1; Form A) from a 50 mg scale salt screen. [Figure 21] FIG. 21 shows the DVS isotherm analysis of compound 1·H2SO4 (1:1; Form A) from the 50 mg scale salt. [Figure 22] FIG. 22 shows the X-ray powder diffraction patterns of compound 1·H2SO4 (1:1; Form A) from a 50 mg scale before and after DVS isotherm analysis. [Diagram 23] FIG. 23 shows the TGA and DSC profiles of compound 1·H3PO4 (1:1; Form A). [Figure 24] FIG. 24 shows the DVS isotherm analysis of compound 1·H3PO4 (1:1; Form A). [Diagram 25] FIG. 25 shows the X-ray powder diffraction patterns of compound 1·H3PO4 (1:1; Form A) before and after DVS. [Figure 26]FIG. 26 shows the TGA and DSC profiles of compound 1·p-toluenesulfonic acid (1:1; Form A). [Figure 27] FIG. 27 shows the DVS isotherm analysis of compound 1·p-toluenesulfonic acid (1:1; Form A). [Figure 28] FIG. 28 shows the X-ray powder diffraction patterns of compound 1·p-toluenesulfonic acid (1:1; Form A) before and after DVS. [Figure 29] FIG. 29 shows the 400 MHz 1H NMR characterization of compound 1·p-toluenesulfonic acid (1:1; Form A) in DMSO-d6. [Diagram 30] FIG. 30 shows the TGA and DSC profiles of compound 1·methanesulfonic acid (1:2; Form A). [Diagram 31] FIG. 31 shows the X-ray powder diffraction pattern of a dry sample of Compound 1·methanesulfonic acid (1:2; Form A). [Diagram 32] FIG. 32 shows the 400 MHz 1H NMR of compound 1·methanesulfonic acid (1:2; Form A) in DMSO-d6. [Diagram 33] FIG. 33 shows the X-ray powder diffraction patterns of wet samples of the polymorph screen derived from stirring 100 μl slurries of Compound 1·HCl (1:1; Form A) in various solvents at 40° C. for 1 day. [Diagram 34] FIG. 34 shows X-ray powder diffraction patterns of wet samples of Compound 1·H2SO4 (1:1) polymorph derived from stirring 100 μl of slurries of Form A in various solvents at 40° C. for 1 day. [Diagram 35] Figure 35 shows the X-ray powder diffraction patterns of dried samples of compound 1·H2SO4 (1:1) polymorphic forms A, C and D derived from stirring slurries of form A in 100 μl of various solvents for 1 day at 40° C. Form B was converted to form A after heating at 40° C. [Diagram 36]Figure 36 shows the X-ray powder diffraction patterns of dried samples of novel crystalline polymorphic forms B, C, E and F of compound 1·H2SO4 (1:1) derived from stirring slurries of form A in 100 μl of various solvents for 1 day at 40° C. Form D is amorphous and was isolated after evaporation of EtOH:H2O (3:1) for several hours. [Figure 37] Figure 37 shows the X-ray powder diffraction patterns of wet samples from a polymorph screening study for compound 1·p-toluenesulfonic acid derived from stirring slurries of Form A in 100 μl of various solvents for 1 day at 40° C. Form C is mostly amorphous and was isolated after evaporation of EtOH:H2O (3:1) for several hours. [Figure 38] Figure 38 shows X-ray powder diffraction patterns of dried samples from a polymorph screening study for compound 1·p-toluenesulfonic acid derived from stirring slurries of Form A in 100 μl of various solvents for 1 day at 40° C. Form C is mostly amorphous and was isolated after evaporation of EtOH:H2O (3:1) for several hours. [Figure 39] FIG. 39 shows the DSC profile of compound 1·p-toluenesulfonic acid (1:1; Form B). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0035] Detailed Description of the Invention In one embodiment, the present invention provides the free base of Compound 1 in various solid states: [ka]

[0036] Compound 1 free base exists in one or more crystalline forms, designated as crystalline Form A, Form B, Form C, Form D, Form E, and Form F, respectively.

[0037] In one embodiment, the present invention provides a crystalline form of Compound 1 free base (Form A), which exhibits an X-ray powder diffraction pattern containing characteristic peaks at diffraction angles 2θ of about 5.9°, 11.9°, and 25.6°, each ±0.2°.

[0038] In one embodiment, the X-ray powder diffraction pattern of crystalline Form A of Compound 1 free base contains characteristic peaks, expressed in units of interplanar distance, at 14.9A, 7.5A, and 3.5A.

[0039] In one embodiment, the X-ray powder diffraction pattern of Compound 1 free base in crystalline form A contains characteristic peaks at diffraction angles 2θ of about 5.6°, 5.9°, 11.9°, 16.5°, 25.6° and 26.2°, each ±0.2°.

[0040] In one embodiment, the X-ray powder diffraction pattern of crystalline form A of Compound 1 free base contains characteristic peaks, expressed in units of interplanar distance, at 15.7A, 14.9A, 7.5A, 5.4A, 3.5A, and 3.4A.

[0041] In one embodiment, the X-ray powder diffraction pattern of crystalline Form A of Compound 1 free base is substantially as shown in FIG. 2A.

[0042] The X-ray diffraction patterns depicted in FIG. 2A are summarized in FIG. 2B.

[0043] In one embodiment, crystalline form A of Compound 1 free base has the TGA profile shown in FIG.

[0044] In one embodiment, crystalline form A of Compound 1 free base has the DSC profile shown in FIG.

[0045] In one embodiment, crystalline form A of Compound 1 free base has a DSC profile with characteristic peaks at 100.3°C, 124.3°C, and 166.3°C.

[0046] In one embodiment, crystalline form A of Compound 1 free base has a melting point with an onset temperature of about 162.3°C.

[0047] In another embodiment, the present invention provides a crystalline form of Compound 1 free base (Form B), which exhibits an X-ray powder diffraction pattern comprising characteristic peaks at diffraction angles 2θ of about 4.7°, 11.2°, 14.1°, 15.3° and 21.2°, each ±0.2°.

[0048] In one embodiment, the X-ray powder diffraction pattern of crystalline form B of Compound 1 free base contains characteristic peaks, expressed in units of interplanar distance, at 18.7A, 7.9A, 6.3A, 5.8A, and 4.2A.

[0049] In one embodiment, crystalline form B of Compound 1 free base has a TGA profile substantially as shown in FIG.

[0050] In one embodiment, crystalline form B of Compound 1 free base has a DSC profile substantially as shown in FIG.

[0051] In one embodiment, crystalline form B of Compound 1 free base has a DSC profile with characteristic peaks at 61.5°C, 118.8°C, and 166.8°C.

[0052] In one embodiment, crystalline form B of Compound 1 free base has a melting point with an onset temperature of about 165.4°C.

[0053] In another embodiment, the present invention provides a method for preparing crystalline form B of Compound 1 free base, comprising the steps of slurrying crystalline form A of Compound 1 free base in ethanol, isopropanol, or a mixed solvent of ethanol:water (3:1) at about 50° C. for about 24 hours, as shown in Table 1; and isolating the resulting crystalline form B.

[0054] In another embodiment, the present invention provides a crystalline polymorph of Compound 1 free base (Form C), exhibiting an X-ray powder diffraction pattern comprising characteristic peaks at diffraction angles 2θ of about 5.7°, 6.4°, 9.2°, 12.8°, 15.7° and 20.7°, each ±0.2°.

[0055] In one embodiment, the X-ray powder diffraction pattern of crystalline form C of Compound 1 free base contains characteristic peaks, expressed in units of interplanar distance, at 15.5A, 13.8A, 9.6A, 6.9A, 5.6A, and 4.3A.

[0056] In one embodiment, the X-ray powder diffraction pattern of crystalline form C of Compound 1 free base contains characteristic peaks at diffraction angles 2θ of about 6.4°, 12.8° and 20.7°, each ±0.2°.

[0057] In one embodiment, the X-ray powder diffraction pattern of crystalline form C of Compound 1 free base contains characteristic peaks, expressed in units of interplanar distance, at 113.8 A, 6.9 A, and 5.6 A.

[0058] In one embodiment, crystalline form C of Compound 1 free base is characterized by a TGA profile substantially as shown in FIG.

[0059] In one embodiment, crystalline form C of Compound 1 free base is characterized by a DSC profile substantially as shown in FIG.

[0060] In one embodiment, crystalline form C of Compound 1 free base is characterized by a DSC profile with a characteristic peak at 162.8°C.

[0061] In one embodiment, crystalline form C of Compound 1 free base has a melting point with an onset temperature of about 160.9°C.

[0062] In another embodiment, the present invention provides a method for preparing crystalline form C of Compound 1 free base, comprising the steps of: slurrying crystalline form A of Compound 1 free base in acetone at about 50° C. for about 24 hours, as shown in Table 1; and isolating the resulting crystalline form.

[0063] In another embodiment, the present invention provides a crystalline polymorph of Compound 1 free base (Form D), characterized by an X-ray powder diffraction pattern substantially as shown in FIG. 10 , including a diffraction pattern comprising characteristic peaks at diffraction angles 2θ of about 4.8°, 8.3°, 10.5°, 11.5°, 14.0°, 18.2°, 20.0°, 23.5°, and 28.5°.

[0064] In one embodiment, crystalline form D of Compound 1 free base is characterized by a TGA profile substantially as shown in FIG.

[0065] In one embodiment, crystalline form D of Compound 1 free base is characterized by a DSC profile substantially as shown in FIG.

[0066] In one embodiment, crystalline form D of Compound 1 free base is characterized by a DSC profile with characteristic peaks at 55.9°C, 118.8°C, and 168.4°C.

[0067] In one embodiment, crystalline form D of Compound 1 free base is characterized by the dynamic water vapor sorption isotherm analysis shown in FIG.

[0068] In one embodiment, the present invention provides a method for preparing a crystalline form of Compound 1 free base, comprising dissolving or slurrying Compound 1 in one or two crystallization solvents selected from the group consisting of alcohols, ketones, esters, ethers, aromatic hydrocarbons, nitriles, halogenated hydrocarbons, and water, with or without seeding, by a crystallization method selected from slurrying, evaporation of solvent, addition of anti-solvent, and / or cooling, and combinations thereof.

[0069] In some embodiments, the controlled or slow addition of an anti-solvent such as water and / or gradual cooling can help control the formation of the crystalline form.

[0070] In some embodiments, seeding can aid or promote the formation of a particular desired crystal form. All of these or other techniques known to those skilled in the art can be combined to achieve the desired result. For example, in some embodiments, such a crystallization process can begin with dissolving Compound 1 in an organic solvent, followed by the addition of anti-solvent, with or without seeding, and cooling.

[0071] In another embodiment, the present invention provides a method for preparing a crystalline form of Compound 1 free base in crystalline form D, as shown in Table 1, comprising the steps of: slurrying crystalline form A of Compound 1 free base in methanol at room temperature for about 24 hours; and isolating the resulting crystalline form.

[0072] The present invention further provides a crystalline salt form of Compound 1.

[0073] The present invention provides a salt of Compound 1 comprising a compound of Formula 1 and an acid, where the acid is selected from HCl, H2SO4, H3PO4, methanesulfonic acid, and p-toluenesulfonic acid.

[0074] In one embodiment, the present invention also provides a method for preparing a crystalline form of a pharma- ceutically acceptable salt of Compound 1, comprising mixing Compound 1 with a suitable acid, e.g., HCl, H2SO4, H3PO4, methanesulfonic acid, and p-toluenesulfonic acid, in one or two crystallization solvents selected from the group consisting of alcohols, ketones, esters, ethers, aromatic hydrocarbons, nitriles, halogenated hydrocarbons, and water, with or without seeding, by a crystallization method selected from slurrying, solvent evaporation, anti-solvent addition, and / or cooling, and combinations thereof, and stirring the mixture.

[0075] In some embodiments, seeding can aid or promote the formation of a particular desired crystal form. For example, in some embodiments, such a crystallization process can begin with dissolving Compound 1 in an organic solvent at elevated temperature, followed by addition of an anti-solvent, with or without seeding, and cooling.

[0076] In some embodiments, seeding may aid or promote the formation of a particular desired crystal form. For water-soluble salts, some non-polar organic solvents, such as aliphatic hydrocarbons, may act as anti-solvents. All of these or other techniques known to those skilled in the art may be combined to achieve the desired result.

[0077] In one embodiment, the present invention provides a crystalline hydrochloride salt of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0078] In one embodiment, the ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to hydrochloride in the crystalline hydrochloride salt is 1:1.

[0079] In one embodiment, crystalline form A of the hydrochloride salt of Compound 1 has an X-ray powder diffraction pattern comprising characteristic peaks at diffraction angles 2θ of about 4.7°, 9.2°, 11.2°, 19.4° and 28.3°, each ±0.2°.

[0080] In one embodiment, the X-ray powder diffraction pattern of the hydrochloride salt of Compound 1 in crystalline form A is substantially as shown in FIG.

[0081] In one embodiment, the crystalline hydrochloride salt of Compound 1 in crystalline Form A is characterized by a TGA profile substantially as shown in FIG.

[0082] In one embodiment, the crystalline hydrochloride salt of Compound 1 in crystalline Form A is characterized by a DSC profile substantially as shown in FIG.

[0083] In one embodiment, the crystalline hydrochloride salt of Compound 1 in crystalline form A is characterized by a DSC profile with characteristic peaks at 44.8°C and 277.9°C.

[0084] In one embodiment, the crystalline hydrochloride salt of Compound 1 in crystalline form A is characterized by a melting point with an onset temperature of about 275.4°C.

[0085] In one embodiment, the crystalline hydrochloride salt of Compound 1 in crystalline Form A has a dynamic water vapor sorption isotherm analysis substantially as shown in FIG. It is characterized by:

[0086] In another embodiment, the present invention provides a method for preparing a crystalline hydrochloride salt of Compound 1 (Form A), comprising the steps of: (a) adding a solution of HCl / methanol to a mixture of compound of formula 1 (using, but not limited to, Form A) in methanol; (b) stirring at about room temperature for about 24 hours; and (c) isolating the resulting crystalline HCl salt of compound 1; The present invention provides a method comprising:

[0087] In some embodiments, the acid addition salt of Compound 1 is a sulfate salt having a crystalline form selected from the group consisting of Form B, Form C, or Form D, substantially as shown in Figure 35 or 36, and having Compound 1 and H2SO4 in a molar ratio of about 1:1, wherein Form B has an X-ray powder diffraction pattern including any three of characteristic peaks at diffraction angles 2θ of about 4.2°, 4.8°, 7.0°, 9.1°, 10.8°, 11.3°, and 12.0°. wherein Form C is characterized by an X-ray powder diffraction pattern comprising any three of characteristic peaks at diffraction angles 2θ of about 5.0°, 8.6°, 10.2°, 18.3°, and 25.7°; and wherein Form D is characterized by an X-ray powder diffraction pattern comprising any three of characteristic peaks at diffraction angles 2θ of about 8.1°, 9.8°, 12.6°, 15.8°, 16.6°, 21.4°, and 23.2°.

[0088] In another embodiment, the present invention provides a crystalline sulfate salt of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0089] In one embodiment, the ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to sulfate in the crystalline sulfate salt is 1:1.

[0090] In one embodiment, the ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to sulfate in the crystalline sulfate salt is 2:1.

[0091] In some embodiments, the acid addition salt of Compound 1 is a sulfate salt having a molar ratio of about 2:1 between Compound 1 and H2SO4 in a crystalline form selected from the group consisting of Form A, Form B, Form C, Form E, and Form F, substantially as shown in Figure 36, wherein Form A is characterized by an X-ray powder diffraction pattern including peaks characteristic of about 4.3°, 5.4°, and 8.4° diffraction angles 2θ; Form B is characterized by an X-ray powder diffraction pattern including any three of peaks characteristic of about 7.9°, 11.6°, 12.1°, 14.5°, 16.1°, 20.4°, 23.1°, 26.1°, and 27.4° diffraction angles 2θ; and Form C is characterized by an X-ray powder diffraction pattern including any three of peaks characteristic of about 4.6°, 7.5°, 9.3°, 12.3°, and 12.6° diffraction angles 2θ. and, wherein Form E is characterized by an X-ray powder diffraction pattern including any three of characteristic peaks at diffraction angles 2θ of about 4.5°, 5.4°, 10.8°, 14.2°, 16.1°, 18.9°, 22.6°, 23.0°, 23.7°, and 27.5°; and wherein Form F is characterized by an X-ray powder diffraction pattern including any three of characteristic peaks at diffraction angles 2θ of about 4.5°, 7.2°, 10.7°, 15.7°, and wherein form F is characterized by an X-ray powder diffraction pattern comprising any three of characteristic peaks at diffraction angles 2θ of about 7.3°, 8.3°, 9.7°, 10.4°, 12.8°, and 25.6°; and wherein form F is characterized by an X-ray powder diffraction pattern comprising any three of characteristic peaks at diffraction angles 2θ of about 7.3°, 8.3°, 9.7°, 10.4°, 12.8°, and 25.6°.

[0092] In one embodiment, the crystalline sulfate salt of Compound 1 in crystalline form A is characterized by an X-ray powder diffraction pattern comprising characteristic peaks at diffraction angles 2θ of about 4.3°, 5.4° and 8.4°, respectively ±0.2°, and / or is characterized by an X-ray powder diffraction pattern substantially as shown in FIG. 22 (bottom row, before DVS).

[0093] In one embodiment, the crystalline sulfate salt of Compound 1 in crystalline form A is characterized by a DSC and / or TGA profile substantially as shown in FIG.

[0094] In another embodiment, the present invention provides a method for preparing a crystalline sulfate salt of Compound 1 (Form A), comprising the steps of: (a) adding a solution of H2SO4 / methanol to a mixture of a compound of formula 1 (using, but not limited to, Form A) in methanol; (b) stirring at about room temperature for about 24 hours; and (c) isolating the resulting crystalline H2SO4 salt of compound 1; The present invention provides a method comprising:

[0095] In one embodiment, the present invention provides a crystalline phosphate salt of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0096] In one embodiment, the ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to sulfate in the crystalline phosphate is about 1:1.

[0097] In one embodiment, the crystalline phosphate salt of Compound 1 (crystalline Form A) has an X-ray powder diffraction pattern comprising characteristic peaks at diffraction angles 2θ of about 4.7°, 9.4°, 14.2°, 23.0° and 26.7°, each ±0.2°.

[0098] In one embodiment, the X-ray powder diffraction pattern of crystalline Form A of the phosphate salt of Compound 1 is substantially as shown in FIG.

[0099] In one embodiment, the crystalline phosphate salt of Compound 1 (crystalline Form A) is characterized by a TGA profile substantially as shown in FIG.

[0100] In one embodiment, the crystalline phosphate salt of Compound 1 (crystalline Form A) is characterized by a melting point with an onset temperature of 229.1°C.

[0101] In one embodiment, the crystalline phosphate salt of Compound 1 (crystalline Form A) is characterized by a DSC profile substantially as shown in FIG.

[0102] In one embodiment, the crystalline phosphate salt of Compound 1 (crystalline Form A) is characterized by a DSC profile with peaks at about 47.8°C and 234.7°C.

[0103] In one embodiment, the crystalline phosphate salt of Compound 1 (crystalline Form A) is characterized by a dynamic water vapor sorption isotherm analysis substantially as shown in FIG.

[0104] In another embodiment, the present invention provides a method for preparing a crystalline phosphate salt of Compound 1 (Form A), comprising: (a) adding a solution of H3PO4 / methanol to a mixture of a compound of formula 1 (using, but not limited to, Form A) in methanol; (b) stirring at about room temperature for about 24 hours; and (c) isolating the resulting crystalline H3PO4 salt of compound 1; The present invention provides a method comprising:

[0105] In one embodiment, the present invention provides a crystalline tosylate salt of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0106] In one embodiment, the ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to tosylate salt in the crystalline tosylate salt is about 1:1.

[0107] In one embodiment, the crystalline tosylate salt of Compound 1 (crystalline Form A) has an X-ray powder diffraction pattern comprising characteristic peaks at diffraction angles 2θ of about 7.4°, 9.8°, 14.7°, 18.4°, 19.5°, and 28.0°, each ±0.2°.

[0108] In one embodiment, the X-ray powder diffraction pattern of the tosylate salt of Compound 1 in crystalline form A is substantially as shown in FIG.

[0109] In one embodiment, the crystalline tosylate salt of Compound 1 (crystalline Form A) is characterized by a TGA profile substantially as shown in FIG.

[0110] In one embodiment, the crystalline tosylate salt of Compound 1 (crystalline Form A) is characterized by a DSC profile substantially as shown in FIG.

[0111] In one embodiment, the crystalline tosylate salt of Compound 1 (crystalline Form A) is characterized by a DSC profile having a peak at about 230.9° C.

[0112] In one embodiment, the crystalline tosylate salt of Compound 1 (crystalline Form A) is characterized by a melting point with an onset temperature of about 229.5°C.

[0113] In one embodiment, the crystalline tosylate salt of Compound 1 (crystalline Form A) is characterized by a dynamic water vapor sorption isotherm analysis substantially as shown in FIG.

[0114] In another embodiment, the present invention provides a crystalline tosylate salt of Compound 1 (crystalline Form B), characterized by a DSC profile substantially as shown in FIG.

[0115] In one embodiment, the crystalline tosylate salt of Compound 1 (crystalline form B) is characterized by an X-ray powder diffraction pattern comprising any three of characteristic peaks at diffraction angles 2θ of about 7.8°, 10.0°, 15.7°, 16.9°, 19.7°, and 20.6°; and a DSC profile having peaks at 82.2° and 229.0°.

[0116] The present invention also provides a method for preparing a crystalline p-toluenesulfonate salt of Compound 1 (Form A), comprising: (a) adding a solution of p-toluenesulfonic acid / methanol to a mixture of a compound of formula 1 (using, but not limited to, Form A) in methanol; (b) stirring at about room temperature for about 24 hours; and (c) isolating the resulting crystalline p-toluenesulfonate salt of compound 1; The present invention provides a method comprising:

[0117] In one embodiment, the present invention provides a crystalline methanesulfonate salt of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine.

[0118] In one embodiment, the ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to the methanesulfonate salt in the crystalline methanesulfonate salt is about 1:2.

[0119] In one embodiment, the X-ray powder diffraction pattern of the mesylate salt of Compound 1, comprising Compound 1 and the mesylate salt in a molar ratio of about 1:2 in crystalline form A, is substantially as shown in FIG.

[0120] In one embodiment, the methanesulfonate salt of Compound 1, comprising Compound 1 and mesylate in a molar ratio of about 1:2 in crystalline form A, is characterized by a TGA profile substantially as shown in FIG.

[0121] In one embodiment, the methanesulfonate salt of Compound 1, comprising about a 1:2 molar ratio of Compound 1 to the mesylate salt in crystalline form A, is characterized by a DSC profile substantially as shown in FIG.

[0122] In one embodiment, the methanesulfonate salt of Compound 1, comprising about a 1:2 molar ratio of Compound 1 to the mesylate salt in crystalline form A, is characterized by a DSC profile with a peak at about 225.0°C.

[0123] In one embodiment, the methanesulfonate salt of Compound 1, comprising about a 1:2 molar ratio of Compound 1 to the mesylate salt in crystalline form A, is characterized by a melting point with an onset temperature of about 223.4°C.

[0124] In another embodiment, the present invention provides a method for preparing a methanesulfonate salt of Compound 1 (Form A), comprising about a 1:2 molar ratio of Compound 1 and mesylate, comprising the steps of: (a) adding a solution of methanesulfonic acid / methanol to a mixture of a compound of formula 1 (using, but not limited to, Form A) in methanol; (b) stirring at about room temperature for about 24 hours; and (c) isolating the resulting crystalline methanesulfonate salt of Compound 1; The present invention provides a method comprising:

[0125] In another aspect, the present invention provides compound 1 and its salts, particularly HCl, HSO, 4、 A method for producing solid amorphous forms of H3PO4, methanesulfonic acid, and p-toluenesulfonic acid salts is provided.

[0126] In one embodiment, the present invention provides a solid amorphous form of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine sulfate.

[0127] In one embodiment, the molar ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to sulfate salt in the solid amorphous form is 1:1.

[0128] In one embodiment, the molar ratio of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to sulfate salt in the solid amorphous form is 2:1.

[0129] In one embodiment, the present invention provides a method for preparing an amorphous sulfate salt of Compound 1, comprising: (a) adding a solution of H2SO4 / methanol to a mixture of a compound of formula 1 (using, but not limited to, Form A) in methanol; (b) stirring at about room temperature for about 24 hours; and (c) isolating the resulting amorphous H2SO4 salt of compound 1; The present invention provides a method comprising:

[0130] In one embodiment, the present invention provides a solid amorphous form of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine methanesulfonate.

[0131] In one embodiment, the molar ratio of N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine to the methanesulfonate salt in the solid amorphous form is 1:1.

[0132] In another embodiment, the present invention provides a method for preparing an amorphous methanesulfonate salt of Compound 1, comprising the steps of: (a) adding a solution of methanesulfonic acid / methanol to a mixture of compound of Formula 1 (Form A) in methanol; (b) stirring at about room temperature for about 24 hours; and (c) isolating the resulting amorphous methanesulfonate salt of Compound 1; The present invention provides a method comprising:

[0133] All of the crystalline polymorphs of the present invention are substantially pure.

[0134] As used herein, the term "substantially pure" means that at least 85% by weight, preferably at least 95% by weight, and more preferably at least 99% by weight, of the compound of formula 1 or a salt thereof is present in the crystalline forms of the present invention, particularly Form A, Form B, Form C, Form D, Form E, and Form F.

[0135] The major peaks depicted in the X-ray powder diffraction (XRPD) patterns of the above crystalline forms are reproducible and within error limits (the specified value ±0.2° in the case of 2θ or 2-theta values), e.g., a 2θ value of 11.5 indicates that the peak has a value within the range of 11.3° to 11.7°, regardless of whether the terms "about" or "approximately" are used as a modifier.

[0136] In the present invention, "X-ray powder diffraction pattern shown in FIG. 2A" means an X-ray powder diffraction pattern exhibiting a major peak as shown in FIG. 2A, where major peak means a peak having a relative intensity of more than 10%, preferably more than 30%, compared to the highest peak in FIG. 1 (designated as having a relative intensity of 100%). Similarly, in the present invention, an X-ray powder diffraction pattern as shown in Figure 10, 11, 12, 13, 14, 18, 22, 25, 28, 31, 33, 34, 35, 36, 37 or 38 means an X-ray powder diffraction pattern exhibiting a major peak as in Figure 10, 11, 12, 13, 14, 18, 22, 25, 28, 31, 33, 34, 35, 36, 37 or 38, where major peak means a peak with a relative intensity of more than 10%, preferably more than 30%, compared to the highest peak in Figure 10, 11, 12, 13, 14, 18, 22, 25, 28, 31, 33, 34, 35, 36, 37 or 38, respectively (with the relative intensity designated as 100%).

[0137] The present invention also provides methods for preparing crystalline form A, crystalline form B, crystalline form C, crystalline form D, crystalline form E, and crystalline form F. Crystallization of the compounds of the present invention from a suitable solvent system comprising at least one solvent may be achieved by spontaneous precipitation (evaporation), cooling, and / or addition of an anti-solvent (in which the compounds of the present invention have a relatively lower solubility) to achieve supersaturation of the solvent system.

[0138] The present invention also provides a method for preparing a salt form of Compound 1, where the salt can be an HCl, H2SO4, H3PO4, methanesulfonic acid, or p-toluenesulfonic acid salt.

[0139] Crystallization may also be achieved with or without the use of suitable crystallizing seeds for crystallizing the compounds of the invention.

[0140] The present invention also relates to a compound of the formula: [ka] wherein X is a halogen, preferably Br or Cl. The present invention provides a method for producing a compound represented by the formula:

[0141] In one embodiment, a method for preparing crystalline Form A of Compound 1 includes the steps of: (a) reacting a compound of Formula II with a compound of Formula 3 in the presence of a catalyst to obtain a reaction mixture containing Compound 1; (b) working up the reaction mixture to isolate crude Compound 1; (c) dissolving crude Compound 1 in aqueous HCl and washing the aqueous solution with an ester; and (d) basifying the aqueous solution with a base to 7 or greater to precipitate Compound 1 free base and isolate the product Form A of Compound 1 free base.

[0142] For general methods for preparing compound 1, see WO 2016 / 014904 A1, which is incorporated herein by reference in its entirety for all purposes as if the entire publication were set forth herein. A specific embodiment of a method for preparing compound 1 in crystalline form A is described in Example 1.

[0143] In one embodiment, the method for preparing crystalline form B of compound 1 includes the steps of: (a) slurrying the free base compound of formula 1 in ethanol, isopropanol, or a mixed solvent of ethanol:water (3:1 v / v) at elevated temperature for a period of time; (b) cooling the slurry of step (a) to room temperature; and (c) isolating the resulting crystalline form B by filtration; and (d) optionally drying the crystalline form B isolated in step (c). In one embodiment, the free base compound of formula 1 in step (a) is crystalline form A. In one embodiment, the elevated temperature is about 50° C.; in one embodiment, the time of slurrying is about 24 hours.

[0144] In one embodiment, the method for preparing crystalline form C of compound 1 comprises the steps of: (a) slurrying the free base compound of formula 1 in acetone at elevated temperature for a period of time; (b) cooling the slurry of step (a) to room temperature; and (c) isolating the resulting crystalline form C by filtration; and (d) optionally drying the crystalline form C isolated in step (c). In one embodiment, the free base compound of formula 1 in step (a) is crystalline form A. In one embodiment, the elevated temperature is about 50° C.; in one embodiment, the time of slurrying is about 24 hours.

[0145] In one embodiment, the method for preparing crystalline form D of compound 1 comprises the steps of: (a) slurrying the free base compound of formula 1 in acetone at room temperature for a period of time; (b) isolating the resulting crystalline form D by filtration; and (c) optionally drying the crystalline form D isolated in step (c). In one embodiment, the free base of compound of formula 1 in step (a) is crystalline form A. In one embodiment, the slurrying time is about 24 hours.

[0146] In one embodiment, a method for preparing the crystalline hydrochloride salt of Compound 1 in Form A comprises the steps of: (a) stirring a mixture of the free base compound of Formula 1 (Form A) and HCl in methanol at room temperature to obtain a slurry containing the crystalline hydrochloride salt in Form A; and (b) isolating the obtained crystalline HCl salt in Form A of compound of Formula 1.

[0147] In one embodiment, a method for preparing the crystalline sulfate salt of Compound 1 in Form A comprises the steps of: (a) stirring a mixture of the free base compound of Formula 1 (Form A) and sulfuric acid (HSO) in methanol at room temperature to obtain a slurry containing the crystalline sulfate salt in Form A; (b) isolating the obtained crystalline HSO salt of Form A of the compound of Formula 1; and (c) optionally drying the isolated crystalline Form A of the HSO salt.

[0148] In one embodiment, a method for preparing crystalline phosphate salt of Compound 1 in Form A comprises the steps of: (a) stirring a mixture of the free base compound of Formula 1 (Form A) and phosphoric acid (HPO) in methanol at room temperature to obtain a slurry containing crystalline phosphate salt in Form A; (b) isolating the obtained crystalline HPO salt of compound of Formula 1 (Form A); and (c) optionally drying the isolated crystalline Form A of the HPO salt.

[0149] In one embodiment, a method for preparing crystalline p-toluenesulfonate salt of Compound 1 in Form A comprises the steps of: (a) stirring a mixture of the free base compound of Formula 1 (Form A) and p-toluenesulfonic acid in methanol at room temperature to obtain a slurry containing crystalline p-toluenesulfonate salt of Form A; (b) isolating the obtained crystalline p-toluenesulfonate salt of compound of Formula 1 (Form A); and (c) optionally drying the isolated crystalline Form A of the p-toluenesulfonate salt.

[0150] In one embodiment, a method for preparing the crystalline methanesulfonate salt of Compound 1 in Form A comprises the steps of: (a) stirring a mixture of the free base compound of Formula 1 (Form A) and methanesulfonic acid in methanol at room temperature to obtain a slurry containing the crystalline methanesulfonate salt of Form A; (b) isolating the obtained crystalline methanesulfonate salt of compound of Formula 1 (Form A); and (c) optionally drying the isolated crystalline Form A of the methanesulfonate salt.

[0151] In one embodiment, a method for preparing an amorphous sulfate salt of Compound 1 includes the steps of: (a) stirring an approximately 2:1 molar ratio mixture of the free base compound of Formula 1 (Form A) and H2SO4 acid in an organic solvent (e.g., methanol) at high heat or room temperature; and (b) removing the solvent (e.g., methanol) by evaporation or centrifugation to dryness to obtain an amorphous H2SO4 acid salt of the compound of Formula 1 in an approximately 2:1 molar ratio.

[0152] In one embodiment, a method for preparing an amorphous sulfate salt of Compound 1 includes the steps of: (a) stirring an approximately 1:1 molar ratio mixture of the free base compound of Formula 1 (Form A) and H2SO4 acid in an organic solvent (e.g., methanol) at high heat or room temperature; and (b) removing the solvent (e.g., methanol) by evaporation or centrifugation to obtain an approximately 1:1 molar ratio amorphous H2SO4 acid salt of the compound of Formula 1.

[0153] In one embodiment, a method for preparing an amorphous phosphate salt of Compound 1 includes the steps of: (a) stirring an approximately 1:1 molar ratio mixture of the free base compound of Formula 1 (Form A) and HPO acid in an organic solvent (e.g., methanol) at high heat or room temperature; and (b) removing the solvent (e.g., methanol) by evaporation or centrifugation to obtain an amorphous HPO acid salt of the compound of Formula 1 in an approximately 1:1 molar ratio.

[0154] In one embodiment, a method for preparing the amorphous methanesulfonate salt of Compound 1 includes the steps of: (a) stirring an about 1:1 molar ratio mixture of the free base compound of Formula 1 (Form A) and methanesulfonic acid in an organic solvent (e.g., methanol) at high heat or room temperature; and (b) removing the solvent (e.g., methanol) by evaporation or centrifugation to dryness to obtain an about 1:1 molar ratio amorphous methanesulfonate salt of compound of Formula 1.

[0155] The invention also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more crystalline polymorphs of crystalline Form A, crystalline Form B, crystalline Form C, crystalline Form D, or crystalline Form E of the compound of Formula 1 and a pharma- ceutically acceptable excipient, adjuvant, or carrier.

[0156] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of one or more salts of the compound of formula 1 and a pharma- ceutically acceptable excipient, adjuvant or carrier. In one embodiment, the pharmaceutical composition is used for oral administration. In a preferred embodiment, the composition is used in a tablet or capsule. Wherein the pharmaceutical composition comprises 1 wt%-99 wt% of the crystalline polymorph of the compound of formula 1 in crystalline form A, crystalline form B, crystalline form C, crystalline form D, or crystalline form E, or more preferably, 1 wt%-70 wt% of the crystalline polymorph, or more preferably, 1 wt%-30 wt% of the compound of formula 1.

[0157] Additionally, the present invention provides methods of inhibiting a cyclin dependent kinase in a subject by administering to the subject a crystalline polymorph of the compound of formula 1 in crystalline form A, crystalline form B, crystalline form C, crystalline form D, or crystalline form E. Preferably, the cyclin dependent kinase is CDK4 or CDK6.

[0158] In another embodiment, a method is provided for treating or preventing cancer or an inflammation-related condition in a subject by administering to the subject a crystalline polymorph of the compound of formula 1 of crystalline form A, crystalline form B, crystalline form C, crystalline form D, or crystalline form E.

[0159] Additionally, the present invention provides a method of inhibiting a cyclin-dependent kinase in a subject by administering to the subject a salt of a compound of formula 1, wherein the salt is selected from HCl, H2SO4, H3PO4, methanesulfonic acid, and p-toluenesulfonic acid salts.

[0160] In one embodiment, the method provides a method of treating CDK4 or CDK6-associated inflammation or cancer, wherein the cancer is selected from the group consisting of brain cancer, metastatic brain cancer, squamous cell carcinoma, bladder cancer, gastric cancer, pancreatic cancer, liver cancer, glioblastoma multiforme breast cancer, head cancer, neck cancer, esophageal cancer, prostate cancer, colorectal cancer, lung cancer, renal cancer, kidney cancer, ovarian cancer, gynecological cancer, thyroid cancer, non-small cell lung cancer (NSCLC), refractory ovarian cancer, or head and neck cancer. In a particular embodiment, the cancer is glioblastoma or metastatic brain cancer.

[0161] In another aspect, the present invention provides the use of either a crystalline or amorphous form of the compound of formula 1 in the manufacture of a medicament for the treatment of CDK4 or CDK6 associated inflammation or cancer, wherein the cancer is selected from the group consisting of brain cancer, metastatic brain cancer, squamous cell carcinoma, bladder cancer, gastric cancer, pancreatic cancer, liver cancer, glioblastoma multiforme breast cancer, head cancer, neck cancer, esophageal cancer, prostate cancer, colorectal cancer, lung cancer, renal cancer, kidney cancer, ovarian cancer, gynecological cancer, thyroid cancer, non-small cell lung cancer (NSCLC), refractory ovarian cancer, or head and neck cancer. In certain embodiments, the cancer is glioblastoma or metastatic brain cancer.

[0162] The terms of the present invention, unless specifically defined, take their ordinary meanings as would be understood by one of ordinary skill in the art.

[0163] For example, the terms "alcohol", "alcohol solvent" and the like refer to C1-C6 alkyl alcohols, preferably C1-C4 alkyl alcohols, such as, in some embodiments, preferably methanol, ethanol, isopropanol, and the like.

[0164] The terms "ketone", "alkyl ketone" and the like refer to a C3-C7 alkanone having the formula RCOR', where R and R' are each independently C1-C4 alkyl, such as, in some embodiments, acetone, butanone, 2-pentanone, 3-pentanone, methyl isobutyl ketone (MIBK), and the like.

[0165] Terms like "ester" refer to lower alkyl fatty acid esters having the formula RCOOR', where R and R' are each independently C1-C4 alkyl, e.g., in some embodiments and preferably, ethyl acetate, ethyl propionate, methyl acetate, propyl acetate, isopropyl acetate, and the like.

[0166] Terms such as "ether" refer to lower alkyl ethers or cyclic ethers (wherein each alkyl has 1 to 4 carbon atoms), including, but not limited to, diethyl ether, diisopropyl ether, ethyl propyl ether, methyl t-butyl ether (MTBE), tetrahydrofuran (THF), 1,4-dioxane, and the like.

[0167] "Aromatic hydrocarbon" and like terms refer to benzene optionally substituted with one to three methyl or ethyl groups, e.g., in some embodiments and preferably, toluene, 1,2-xylene, 1,4-xylene, 1,3-xylene, cumene, ethylbenzene, and the like.

[0168] The term "halogenated hydrocarbon" and the like means a C1-C6 alkane substituted with 1-6, preferably 1-4, F and / or Cl atoms, e.g., in some embodiments, preferably dichloromethane, chloroform, 1,1,1-trifluoroethane, etc. The term "nitrile", "nitrile solvent" and the like means a C2-C4 alkylnitrile, i.e., CH3CN, CH3CH2CN, CH3CH2CH2CN, or CH3CH(CN)CH3, preferably CH3CN.

[0169] As used herein, the term "room temperature" means a temperature in the range of about 20 to about 25°C, with an average of about 23°C, unless otherwise specified.

[0170] Either the crystalline or amorphous form of the compound free base or a salt thereof, particularly the former, may exist in a solvated form, which can be readily determined by one of ordinary skill in the art through a variety of conventional analytical methods. Such solvates of the solid forms are included in the present invention.

[0171] The term "solvate" as used herein refers to a physical association of a compound of the present invention with a stoichiometric or non-stoichiometric amount of solvent molecules. For example, one molecule of the compound is associated with one or more, preferably 1-3, solvent molecules. It is also possible for multiple (e.g., 1.5 or 2) molecules of the compound to share one solvent molecule. This physical association may include hydrogen bonding. In certain instances, the solvate will be capable of being isolated as a crystalline solid. The solvent molecules in the solvate may be present in an ordered and / or disordered arrangement. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Methods of solvation are generally known in the art.

[0172] The term "treatment" or "treating" means the management and care of a patient for the purpose of combating a disease, condition or disorder.

[0173] The term "therapeutically effective amount" means an amount of a drug or therapeutic agent that elicits the desired biological and / or medical response in a tissue, system, or animal (including humans) that is being sought by a researcher or clinician.

[0174] The term "subject" or "patient" means a mammal.

[0175] The term "mammal" or "mammals" includes, but is not limited to, humans, dogs, cats, horses, pigs, cows, monkeys, rabbits, and mice. A preferred mammal is a human.

[0176] The term "administer" means to apply the compound of the present invention, or its pharma- ceutically acceptable salt, prodrug, or composition, to a subject in need of treatment. The administration of the composition of the present invention to carry out the treatment method of the present invention is carried out by administering a therapeutically effective amount of the compound in the composition to a subject in need of such treatment or prophylaxis. The need for prophylactic administration according to the method of the present invention is determined by using well-known risk factors. The effective amount of each compound is, in the final analysis, determined by the physician in charge of the case, but will vary depending on factors such as the exact disease being treated, the severity of the disease, and other diseases or conditions the patient suffers from, the route of administration selected, other drugs and treatments that the patient may concomitantly require, and other factors in the physician's judgment.

[0177] As used herein, the term "pharmacologically acceptable" means those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for contact with the tissues of mammals, particularly humans, without undue toxicity, irritation, allergic response and other problems, commensurate with a reasonable benefit / risk ratio.

[0178] The term "therapeutically effective amount" as used herein means an amount of a compound that, when administered to a subject to treat a disease, or at least one of the clinical symptoms of a disease or disorder, is sufficient to affect such treatment for the disease, disorder, or condition. A "therapeutically effective amount" may vary depending on the compound, the disease, disorder, and / or symptoms of the disease or disorder, the severity of the disease or disorder, and / or symptoms of the disease or disorder, the age of the subject being treated, and / or the weight of the subject being treated. The appropriate amount in any given case will be apparent to one of skill in the art or can be determined by routine experimentation. In the case of a combination therapy, a "therapeutically effective amount" means the total amount of the combination for effective treatment of a disease, disorder, or condition.

[0179] The pharmaceutical composition comprising the compound of the present invention can be administered to a subject in need of treatment via oral, inhalation, rectal, parenteral or topical administration. For oral administration, the pharmaceutical composition can be a conventional solid formulation such as tablet, powder, granule, capsule, liquid formulation such as water or oil suspension, or other liquid formulation such as syrup, solution, suspension; for parenteral administration, the pharmaceutical composition can be a solution, aqueous solution, oil suspension concentrate, freeze-dried powder, etc. In one embodiment, the pharmaceutical composition formulation is selected from tablet, coated tablet, capsule, suppository, nasal spray or injection, more preferably tablet or capsule. The pharmaceutical composition can be a single dose with a precise dosage. In addition, the pharmaceutical composition can further comprise additional active ingredients.

[0180] All formulations of the pharmaceutical composition of the present invention can be prepared by conventional methods in the pharmaceutical field. For example, the active ingredient can be mixed with one or more excipients and then made into a desired formulation. "Pharmaceutically acceptable carrier" refers to conventional pharmaceutical carriers suitable for the desired formulation, such as diluents, vehicles such as water, various organic solvents, fillers such as starch, sucrose, binders such as cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone (PVP), wetting agents such as glycerol, disintegrants such as agar, calcium carbonate and sodium bicarbonate, absorption enhancers such as quaternary ammonium compounds, surfactants such as hexadecanol, absorption carriers such as kaolin and soap clay, lubricants such as talc, calcium, stearate, magnesium stearate, polyethylene glycol, etc. In addition, the pharmaceutical composition further comprises other pharmaceutically acceptable excipients such as dispersants, stabilizers, thickeners, complexing agents, buffers, penetration enhancers, polymers, aromatic compounds, sweeteners, and dyes. In one embodiment, the excipients are suitable for the desired formulation and administration type. The term "disease" or "disorder" or "condition" means any disease, ailment, illness, symptom or sign.

[0181] It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. In general, when either the singular or plural form of a noun is used, it refers to both the singular and the plural of the noun.

[0182] The term "about" or "approximately" generally includes up to plus or minus 10% of the indicated number unless otherwise defined. For example, "about 10%" can indicate a range of 9% to 11%, and "about 20" can mean 18 to 22. Preferably, "about" includes values ​​up to plus or minus 6% of the indicated value. Alternatively, "about" includes values ​​up to plus or minus 5% of the indicated value. Other meanings of "about" may be apparent from the context, such as rounding, so for example, "about 1" may also mean 0.5 to 1.4.

[0183] However, in the case of melting or onset temperatures of crystalline forms measured by DSC thermograms, the term "about" indicates that the melting or onset temperature may vary, typically within ±2° C., regardless of the absolute value of the melting or onset temperature, as will be understood by those of skill in the art. When a parameter is not critical, as will be understood by those of skill in the art, the number is often given for illustrative purposes only, not limiting.

[0184] The terms "including," "comprising," "containing," or "having" and variations thereof are meant to encompass the items listed thereafter and equivalents thereof, as well as additional subject matter, unless expressly stated otherwise. The following non-limiting examples further illustrate certain aspects of the present invention.

[0185] Working Example The present invention is further illustrated, but not limited, by the following examples which illustrate the invention, in which the techniques or methods are conventional techniques or methods in the art unless otherwise indicated. EXAMPLES

[0186] Synthesis of Compound 1 (Form A) compound 1 N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine (1) [ka]

[0187] A mixture of 5-(2-chloro-5-fluoropyrimidin-4-yl)-2-methyl-3-(propan-2-yl)-2H-indazole (2) (20.7 mol, 1.0 equiv.), 5-((4-ethylpiperazin-1-yl)methylpyridin-2-amine (3) (24.5 mol, 1.2 equiv.), K3PO4·3H2O (69.5 mol, 3.4 equiv.), Pd2(dba)3 (0.97 mol, 0.05 equiv.) and Xantphos (1.02 mol, 0.05 equiv.) in 184 L of 1,4-dioxane was degassed with N2 and then heated at 90-100 °C with stirring for 24 h. The mixture was cooled to 40-50 °C and added with L-cysteine ​​(0.05 equiv.) and water (6.8 L) and then stirred at 40-50° C. for 3-5 hours. The mixture was cooled to 0-10° C., stirred for 1-3 hours, centrifuged, and then washed with fresh water. The wet cake was dissolved in 1N aqueous HCl, washed twice with isopropyl acetate, and decolorized with activated charcoal. The solution was adjusted to pH 9-10 with 17% aqueous Na2CO3 and centrifuged. The wet cake was washed three times with water and dried to give 4.9 kg (47%) of the free base of 1 (Form A) as an off-white solid (purity: 99.4%); N-(5-((4-ethylpiperazin-yl) methyl) pyridin-2-yl-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl) pyrimidin-2-amine (1). MS m / z MH+ 489; 1 H NMR (300 MHz, CDCl3, ppm): δ 0.97 (t, 3H, J = 7.1 Hz), 1.51 (d, 2H, J = 7.0 Hz), 2.27-2.51 (overlapping m, 10 H), 3.33(s, 3H), 3.54-3.68(m, 1H), 4.15(s, 3H), 7.64-8.71(overlapping m, 7H), 10.00(s, 1H).

[0188] The free base of 1 (form A) was characterized by polarized light microscopy (PLM), X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA); Figures 1-3. As shown in the PLM and XRPD results (Figures 1 and 2), it is crystalline with irregular small particles (Figures 1 and 2). The DSC profile (Figure 3) shows that there are three endothermic peaks with an onset of the melting point at 162.3 °C (54.5 J / g). The TGA profile shows that there is a weight loss of 0.64% from room temperature to 120 °C.

[0189] XRPD conditions for all samples are given below: SCAN:4.0 / 39.9779 / 0.01972 / 18.6(sec), Cu(40kV,40mA); PEAK: Parabolic filter, Peak-Top=Summit; NOTE: Intensity = counts, 2T(0) = 0.0(°), Calculate wavelength d-Spacing = 1.54056A(Cu / K-alpha1). EXAMPLES

[0190] Free Base Polymorph Screening for Compound 1: The free base of Compound 1 (Form A) was placed into each of five glass vials, followed by the addition of solvent to each vial as described in Table 1. If a clear solution was obtained, then 1 was added until a suspension was formed. Otherwise, the suspension was stirred at 50° C. for 24 hours. The resulting solids were examined by XRPD and then dried under vacuum at 40° C. overnight. The dried samples were also analyzed by XRPD. See FIG. 4.

[0191] The three new crystalline free base forms obtained (Forms B, C and D) are summarized in Table 1. The original Form A converted to Form B in isopropanol, ethanol and a 3:1 ethanol:water mixture, while in acetone it converted to Form C. Form D was obtained from methanol.

[0192] Table 1 Free base polymorph screening of compound 1 (form A) by slurry method [Table 1]

[0193] Compound 1 Free Base (Form B) The free base of Compound 1 (Form B) was obtained in IPA, or ethanol and ethanol / water in a 3:1 ratio. The DSC profile showed three endothermic peaks and a melting point of 165.4° C. The TGA profile showed a weight loss of 0.43%, which may be due to the release of solvent ( FIG. 6 ).

[0194] Compound 1 Free Base (Form C) The free base of 1 (Form C) was obtained in acetone. Based on the TGA and DSC profiles (FIG. 7), the melting point has an onset of 160.9° C. (103.6 J / g) with a weight loss of 0.43% from room temperature to 120° C.

[0195] Compound 1 Free Base (Form D) The free base of 1 (form D) was obtained in methanol. As shown in Figure 8, the DSC profile has three peaks. The first peak has an onset temperature of 55.9 °C (54.6 J / g) with a weight loss of 1.16%, the second peak has an onset temperature of 118.8 °C (13.2 J / g), which may be related to the release of residual solvents. The third peak has a temperature of 168.4 °C (68.5 J / g). Dynamic water vapor sorption (DVS) isotherm analysis (Figure 9) indicates that the free base of 1 (form D) is hygroscopic (5.13% at 80% RH) and there is no change in the crystal morphology after DVS (Figure 10).

[0196] Selected data from Figure 4 are set forth below. Compound 1 free base form B (dry from IPA): [Table 2]

[0197] Compound 1 free base form C (dry form from acetone): [Table 3] EXAMPLES

[0198] Preparation and Characterization of Salt Forms of Compound 1 (50 mg Scale) Approximately 50 mg of free base 1 (Form A) was placed in each of six glass vials and dissolved in 2.0 mL of methanol. An appropriate amount of hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid or p-toluenesulfonic acid in methanol was added to the vial in the molar ratios summarized in Table 2. The mixture was magnetically stirred at room temperature for 24 hours. If a clear solution was obtained, a vacuum centrifugation was performed at 60° C. to evaporate the solvent. Otherwise, the suspension was centrifuged at 10,000 rpm for 5 minutes to separate the remaining solids. The resulting wet solids were analyzed by XRPD (FIG. 11) and then dried under vacuum at 40° C. for 4 hours. The dry samples were subsequently analyzed by XRPD (FIGS. 12 and 13) and the results for both the wet and dry salts are shown in Table 2. XRPD results, at a 50 mg scale, show that the dry hydrochloride (1:1; Form A), phosphate (1:1; Form A), sulfate (1:1; Form A) and tosylate (1:1; Form A) salts are crystalline material, whereas the sulfate (2:1) and methanesulfonate (1:1) salts are amorphous.

[0199] Table 2 50 mg scale salt screen for compound 1 in methanol [Table 4] *ND: Not determined. EXAMPLES

[0200] Preparation and Characterization of Salt Forms of Compound 1 (500 mg Scale) Approximately 500 mg of free base 1 (Form A) was placed in each of six glass vials and dissolved in 20 mL of methanol. A methanolic solution of hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, and p-toluenesulfonic acid, with the molar ratios shown in Table 3, was added to each vial and the mixture was stirred at room temperature for 24 hours. If a clear solution was obtained, the solid material was isolated using vacuum centrifugation at 50° C. for 1.5 hours. Subsequently, 20 mL of MTBE was added to the concentrate and the mixture was stirred for 30 minutes, collected, and dried. If a clear solution was not obtained, the suspension was centrifuged at 10,000 rpm for 5 minutes to separate the remaining solids. The solids were dried under vacuum at 40° C. for 1 day and the dry samples were analyzed by XRPD with the results shown in Table 3.

[0201] Based on the 500 mg scale XRPD diffractograms (FIGS. 14 and 15), the hydrochloride (1:1; Form A), phosphate (1:1; Form A), tosylate (1:1; Form A) and sulfate (2:1; Form A) salts of compound 1 were crystalline, while the (1:1) sulfate salt was amorphous. The 500 mg sulfate results are in contrast to the 50 mg scale results, since at the 50 mg scale, the (1:1; Configuration A) sulfate salt was crystalline (FIG. 13) and the (2:1) sulfate salt was amorphous (FIG. 12). Similarly, the methanesulfonate salt as the (1:1) salt was amorphous at the 50 mg scale and crystalline after isolation as the (1:2; Form A) salt at the 500 mg scale.

[0202] The molar ratios of the isolated salts of compound 1 in Table 3 (1 free base:acid) were confirmed for the hydrochloride (1:1; Form A), phosphate (1:1; Form A), sulfate (1:1; Form A) and sulfate (2:1; Form A) salts by RP-HPLC analysis with evaporative light scattering detection (ELSD; Table 4); 1 1 H NMR spectroscopy confirmed the methanesulfonate salt (1:2; Form A) and the tosylate salt (1:1; Form A).

[0203] Table 3 500 mg scale salt screen for compound 1 in methanol [Table 5]

[0204] Table 4 RP-HPLC with ELSD detection assay conditions [Table 6] EXAMPLES

[0205] DVS, TGA, DSC and 1 H-NMR characterization Compound 1·HCl (1:1; Form A) The TGA and DSC profiles of compound 1·HCl (1:1; Form A) (dry) are shown in Figure 16. The DSC profile shows two endothermic peaks and a melting point with an onset temperature of 275.4 °C (198.5 J / g). The TGA profile shows a weight loss of 2.95% and the DVS isotherm analysis (Figure 17) shows that 1·HCl (1:1; Form A) is hygroscopic (3.64% at 80% RH) and the crystalline form does not change as shown in the XRPD diffractograms before and after DVS (Figure 18). These results indicate that 1·HCl (1:1; Form A) is in monohydrate form (theoretical water content 3.3%, w / w). ... 1 The H NMR spectrum is shown in Figure 19.

[0206] Compound 1·H 2 SO 4 (1:1) The TGA and DSC profiles of crystalline compound 1 H2SO4 (1:1; Form A) at 50 mg scale are shown in Figure 20. The TGA profile shows a weight loss of 4.92% between 31.6 °C and 120 °C, and the DSC profile shows two endothermic peaks, the first with an onset temperature of 34.3 °C (268.1 J / g) and the second with an onset temperature of 136.0 °C (13.2 J / g). DVS isotherm analysis (Figure 21) showed that compound 1 H2SO4 (1:1; Form A) is highly hygroscopic (26.42% at 80% RH) and has some degree of crystal morphology change as shown by XRPD after DVS (Figure 22).

[0207] Compound 1·H 3 PO 4 (1:1;Form A) The DSC and TGA profiles of compound 1·H3PO4 (1:1; Form A) are shown in Figure 23. The DSC profile showed a melting point onset at 229.1 °C (152.2 J / g) and the TGA profile showed a weight loss of 0.59% from 31.4 °C to 120 °C. DVS isotherm analysis (Figure 24) showed that compound 1·H3PO4 (1:1) is hygroscopic (3.16% at 80% RH) and undergoes some change in crystal morphology after DVS as shown by XRPD (Figure 25, with peaks listed below). [Table 7]

[0208] Compound 1·p-toluenesulfonic acid (1:1; Form A) The TGA and DSC profiles of compound 1·p-toluenesulfonic acid (1:1; Form A) are shown in Figure 26. The TGA profile showed a weight loss of 0.53% from room temperature to 120 °C, and the DSC profile showed an endothermic peak with a melting onset temperature of 229.5 °C (85.0 J / g). DVS isotherm analysis (Figure 27) showed that compound 1·p-toluenesulfonic acid (1:1; Form A) is slightly hygroscopic (1.67% at 80% RH) and there was no change in the crystalline morphology after DVS as shown by XRPD (Figure 28).

[0209] Compound 1·methanesulfonic acid (1:2; Form A) The TGA and DSC profiles of compound 1·methanesulfonic acid (1:2; Form A) are shown in Figure 30, and the XRPD pattern is shown in Figure 31. The DSC profile showed an endothermic peak with an onset melting temperature of 223.4 °C (56.3 J / g), and the TGA profile showed a weight loss of 2.48% from 31 °C to 120 °C. 1 The H NMR spectrum showed that the molar ratio of compound 1 to methanesulfonic acid was 1:2 (Figure 32). EXAMPLES

[0210] Polymorphism Screening for Salts of Compound 1 General steps: Approximately 20 mg of each initial salt form of Compound 1 (Form A) was placed into each of nine glass vials and suspended in 100 μL of the various solvents listed in Tables 5-8. If a clear solution was obtained, it was allowed to evaporate for several hours. Otherwise, the slurry was stirred at 40 °C for 24 h. A sample of the slurry was drawn and analyzed by XRPD (wet sample). The remaining solid was heated under reduced pressure at 40 °C overnight and then analyzed by XRPD (dry sample).

[0211] Compound 1·HCl (1:1; Form A) No new crystalline polymorphs derived from compound 1·HCl (1:1; Form A) were formed in any of the listed solvents (Table 5, FIG. 33). Table 5 Polymorphic screening of compound 1·HCl (1:1; Form A) [Table 8] *100 μL of slurries of compound 1·HCl (1:1; Form A) in various solvents were stirred at 40 °C for 1 day; ND: not determined.

[0212] Compound 1 sulfate (1:1; forms B, C, and D) Compound 1 HSO 4(1:1; Form A) three new crystalline forms were obtained by the slurry method: Forms B, C and D (Table 6, Figures 34 and 35). Form B was converted to Form A after heating at 40°C. Table 6 Polymorphic screening of compound 1·H2SO4 (1:1; Form A) [Table 9] *100 μL of slurries of compound 1·H2SO4 (1:1; Form A) in various solvents were stirred at 40 °C for 1 day; ND: not determined.

[0213] Form B (wet from IPA): [Table 10]

[0214] Form C (wet from acetone): [Table 11]

[0215] Form D (wet from THF ): [Table 12]

[0216] Form D (dry from THF): [Table 13]

[0217] Compound 1 sulfate (2:1; forms B, C, D, E and F) Five new forms of 1 H2SO4(2:1) were derived from compound 1 H2SO4(2:1; Form A) and are summarized in Table 7 and Figure 36. Forms B, C, E, and F were crystalline, whereas Form D was amorphous.

[0218] Table 7 Polymorphic screening of compound 1·H2SO4 (2:1; Form A) [Table 14] *100 μL of slurries of compound 1·H2SO4 (2:1; Form A) in various solvents were stirred at 40 °C for 1 day and then allowed to evaporate for several hours.

[0219] Compound 1·H 2 SO 4 (2:1) Form A [Table 15]

[0220] Compound 1·H 2 SO 4 (2:1) Form B (dry from ACN) [Table 16]

[0221] Compound 1·H 2 SO 4 (2:1) Form C (dry from EtOH) [Table 17]

[0222] Compound 1·H 2 SO 4 Form E (2:1) (dry from acetone) [Table 18]

[0223] Compound 1·H 2 SO 4 (2:1) Form F (dry from THF) [Table 19]

[0224] Compound 1·H 2 SO 4 (2:1) Form G (or C) (dry from toluene) [Table 20]

[0225] Compound 1·p-toluenesulfonic acid (1:1; Form B) One new crystalline form (form B) was derived from compound 1·p-toluenesulfonic acid (1:1; form A), whereas form C was mostly amorphous (Table 8 and Figures 37, 38). The DSC profile of compound 1·p-toluenesulfonic acid (1:1; form B) showed two endothermic peaks (Figure 39). The first had an onset temperature of 82.2 °C (9.9 J / g), which could be associated with the solid form transition to form A, and the second could be melting with an onset temperature of 229.0 °C (81.4 J / g).

[0226] Table 8 Polymorphic screening of compound 1·p-toluenesulfonic acid (1:1; Form A) [Table 21] *Slurries of compound 1·p-toluenesulfonic acid (1:1; Form A) in 100 μL of various solvents were stirred at 40 °C for 1 day. Form C is mostly amorphous and was isolated after evaporation of EtOH:H2O (3:1) for several hours. ND: Not determined.

[0227] Compound 1·p-tosylate form A (dry after DVS, see Figure 28) [Table 22]

[0228] Compound 1 p-tosylate form B (wet from THF) [Table 23]

[0229] definition Acetonitrile (ACN) Differential Scanning Calorimetry (DSC) Dynamic Vapor Sorption (DVS) Ethanol (EtOH) Ethyl acetate (EtOAc) Evaporative Light Scattering Detection (ELSD) Isopropyl alcohol (IPA) Methanol (MeOH) Methyl t-butyl ether (MTBE) Nuclear magnetic resonance spectroscopy (NMR) Polarized Light Microscope (PLM) Tetrahydrofuran (THF) Thermogravimetric analysis (TGA) X-ray powder diffraction (XRPD) Table 9 utensils [Table 24]

[0230] All publications cited herein are incorporated by reference in their entirety for all purposes. It should be understood that the embodiments described herein should be considered as examples only, without limiting the scope of the present invention. The description of features or aspects in each embodiment should generally be considered applicable to other similar features or aspects in other embodiments. Although several embodiments have been described in the above examples, it will be understood by those skilled in the art that various changes in form and details can be made without departing from the true spirit and scope of the disclosure as defined by the following claims.

Claims

1. The acid addition salt of the crystalline form of N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl)-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine, comprising the compound N-(5-((4-ethylpiperazin-yl)methyl)pyridin-2-yl)-5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine and hydrochloric acid (HCl) in a 1:1 molar ratio, is characterized by an X-ray powder diffraction pattern containing any three of characteristic peaks at diffraction angles 2θ of 4.7°, 9.2°, 11.2°, 19.5° and 28.3° (Form A).

2. (a) 4.7°, 8°, 9.2°, 11.2°, 11.9°, 13°, 13.8°, 14.4°, 15.2°, 16.1°, 18.4°, 18.9°, 19.5°, 20°, 20.5°, 21°, 21.7°, 22.5°, 23.2°, 23.9°, 24.4°, 24.8°, 25.9°, 27.6°, 28.3°, 29°, 30.5°, 31.5 2. The acid addition salt of claim 1, in a crystalline form (Form A), characterized by either (a) an X-ray powder diffraction pattern containing peaks at diffraction angles 2θ of 32.3°, 33.4°, 34.1°, and 38.1°; or (b) a DSC profile containing characteristic peaks at 44.8° C. and 277.9° C., and having a melting point with an onset temperature of 275.4° C.

3. A pharmaceutical composition comprising an acid addition salt according to any one of claims 1 to 2 and a pharma- ceutically acceptable carrier.

4. 4. The pharmaceutical composition according to claim 3, wherein the composition is in oral dosage form and comprises 1 wt%-70 wt% of the acid addition salt according to any one of claims 1 to 2.

5. A pharmaceutical composition for treating a disease or condition associated with CDK4 and / or CDK6 activity, comprising an acid addition salt according to any one of claims 1 to 2.

6. 6. The pharmaceutical composition of claim 5, wherein the disease or disorder is inflammation or cancer selected from the group consisting of brain tumor, metastatic brain tumor, squamous cell carcinoma, bladder cancer, gastric cancer, pancreatic cancer, liver cancer, glioblastoma multiforme, breast cancer, head cancer, neck cancer, esophageal cancer, prostate cancer, colorectal cancer, lung cancer, renal cancer, kidney cancer, ovarian cancer, gynecological cancer, thyroid cancer, non-small cell lung cancer (NSCLC), refractory ovarian cancer, and head and neck cancer.

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