Crystalline forms of phosphoinositide 3-kinase (PI3K) inhibitors

Abstract

The present invention relates to salts and crystalline forms of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionamide, crystalline forms of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropyl-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, and crystalline forms of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxypropyl-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, which are PI3K inhibitors useful in treating cancer and other diseases.

Description

Technical Field

[0001] The present invention relates to 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionamide, 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxyprop-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide and 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxyprop-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide. 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, which are PI3K inhibitors useful in treating cancer and other diseases. Background Art

[0002] Compounds having formula A are 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, compounds having formula B are 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, and compounds having formula C are 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide. 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide:

[0003]

[0004] It is a phosphoinositide 3-kinase (PI3K) inhibitor that can be used to treat various diseases (including cancer). For drug development, it is usually appropriate to adopt a drug form with ideal properties in terms of its preparation, purification, reproducibility, stability, bioavailability and other characteristics. Therefore, the salts and crystalline forms of the compounds of Formula A, Formula B and Formula C provided herein solve the continued need for the development of PI3K inhibitors for the treatment of serious diseases. SUMMARY OF THE INVENTION

[0006] The present invention provides salts and crystalline forms of a compound of formula A, a crystalline form of a compound of formula B, and a crystalline form of a compound of formula C:

[0007]

[0008] The present invention also provides a composition comprising a salt and a crystalline form of a compound of formula A provided herein and at least one pharmaceutically acceptable carrier. The present invention also provides a composition comprising a crystalline form of a compound of formula B and at least one pharmaceutically acceptable carrier. The present invention also provides a composition comprising a crystalline form of a compound of formula C and at least one pharmaceutically acceptable carrier.

[0009] The invention also provides processes for preparing the salts and crystalline forms of the invention.

[0010] The present invention also provides a method of treating a disease associated with aberrant expression or activity of PI3K kinase in a patient, the method comprising administering to the patient a therapeutically effective amount of a salt or crystalline form of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 Shown is the XRPD pattern of crystalline Form IA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0012] Figure 2 Shown are the results of DSC experiments of crystalline Form IA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0013] Figure 3 Shown are the results of TGA experiments for crystalline Form IA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0014] Figure 4 Shown is the XRPD pattern of crystalline Form IIA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0015] Figure 5 Shown are the results of DSC experiments of crystalline Form IIA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0016] Figure 6 Shown are the results of TGA experiments of crystalline Form IIA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0017] Figure 7 Shown is the XRPD pattern of crystalline Form IIIA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0018] Figure 8 The results of DSC experiments of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide crystalline form IIIA are shown.

[0019] Fig. 9 Shown are the results of TGA experiments of crystalline Form IIIA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0020] Fig.10 Shown is the XRPD pattern of crystalline Form IB of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide.

[0021] Fig.11 Shown are the results of DSC experiments of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide crystalline Form IB.

[0022] Fig.12 Shown is the XRPD pattern of crystalline Form IIB of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide.

[0023] Fig.13 Shown are the results of DSC experiments of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide crystalline Form IIB.

[0024] Fig.14 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide crystalline Form IC XRPD pattern.

[0025] Fig.15 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 DSC experimental results of 5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide crystalline form IC.

[0026] Fig.16A An asymmetric crystalline unit of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide methanol solvate is shown, with thermal ellipsoids drawn at the 30% probability level.

[0027] Fig. 16B A crystalline unit of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide is shown, with thermal ellipsoids drawn at the 30% probability level. DETAILED DESCRIPTION

[0028] The present invention particularly relates to salts and crystalline forms of the PI3K inhibitor 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide having formula A:

[0029]

[0030] They can be used, for example, to prepare solid dosage forms of the above compounds for use in treating various diseases, including cancer.

[0031] The present invention also relates in particular to a crystalline form of the PI3K inhibitor 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide having formula B:

[0032]

[0033] The present invention also particularly relates to a PI3K inhibitor having the formula C: 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3Crystalline forms of 5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide:

[0034]

[0035] Generally, about for example hygroscopicity, solubility, stability etc., different crystalline forms of the same substance have different overall properties. The form with a high melting point has good thermodynamic stability usually, which is conducive to extending the shelf life of the pharmaceutical preparation containing the solid form. The form with a lower melting point is usually less stable thermodynamically, but is advantageous in that they have increased water solubility, thereby converting into increased drug bioavailability. Weakly hygroscopic forms are more ideal because of their thermal stability and wet stability, and resist degradation during long-term storage. Anhydrous forms are usually more ideal because they can be manufactured consistently without worrying about causing weight or composition changes due to solvent or water content changes. On the other hand, hydrated forms or solvated forms may be advantageous because they are unlikely to be hygroscopic and may show improved humidity stability under storage conditions.

[0036] As used herein, "crystalline form" is intended to refer to a certain lattice conformation of a crystalline substance. Different crystalline forms of the same substance typically have different lattices (e.g., unit cells) due to the different physical properties unique to each crystalline form. In some cases, different lattice conformations have different water or solvent contents. Different crystal lattices can be identified by solid-state characterization methods, such as by X-ray powder diffraction (XRPD). Other characterization methods, such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor adsorption (DVS), solid-state NMR, etc., also help identify the crystalline form and help determine stability and solvent / water content.

[0037] Crystalline forms of substances include solvated (e.g., hydrated) forms and non-solvated (e.g., anhydrous) forms. A hydrated form is a crystalline form that includes water in the crystal lattice. The hydrated form can be a stoichiometric hydrate, in which water is present in the lattice at a certain water / molecule ratio, such as a hemihydrate, a monohydrate, a dihydrate, etc. The hydrated form can also be non-stoichiometric, in which the water content is variable and depends on external conditions, such as humidity.

[0038] Crystalline forms are most often characterized by XRPD. The XRPD pattern of reflection (peak) is generally regarded as the fingerprint of a specific crystalline form. It is well known that the relative intensity of XRPD peaks can vary widely depending on sample preparation techniques, crystal size distribution, filters, sample fixing procedures, and the specific instrument used. In some cases, new peaks may be observed or existing peaks may disappear, depending on instrument type or setting (e.g., whether Ni filters are used). As used herein, the term "peak" refers to a reflection with a relative height / intensity of at least about 4% of the maximum peak height / intensity. In addition, instrument changes and other factors may affect 2θ values. Therefore, peak designations, such as those reported herein, may vary plus or minus about 0.2 ° (2θ), and the term "substantially" as used herein in the context of XRPD is intended to cover the above-mentioned changes.

[0039] Likewise, temperature readings associated with DSC, TGA, or other thermal experiments may vary by about ±4°C, depending on the instrument, specific settings, sample preparation, etc. For example, in the case of DSC, it is known that the observed temperature will depend on the rate of temperature change as well as the sample preparation technique and the specific instrument employed. Thus, as noted above, the values ​​reported herein for DSC thermograms may vary by ±4°C. Thus, crystalline forms reported herein as having a DSC thermogram "substantially" as shown in any of the accompanying drawings should be understood to include such variations.

[0040] Crystalline form of the compound of formula A

[0041] The compound of formula A can be isolated in a variety of crystalline forms, including, for example, anhydrous and / or non-solvated or solvated crystalline forms. In some embodiments, the crystalline form of the compound of formula A is solvated. In some embodiments, the crystalline form of the compound of formula A is anhydrous. In some embodiments, the crystalline form of the compound of formula A is non-solvated. In some embodiments, the crystalline form of the compound of formula A is anhydrous and non-solvated. "Anhydrous" means that the crystalline form of the compound of formula A is substantially free of bound water in the crystal lattice structure, that is, the compound does not form a crystalline hydrate.

[0042] In some embodiments, the present application provides a method for preparing a crystalline form of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide. In some embodiments, the method includes dissolving 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide in a solvent to form a mixture, and crystallizing the compound from the mixture.

[0043] In some embodiments, the solvent comprises isopropyl acetate. In some embodiments, the solvent further comprises heptane.

[0044] In some embodiments, the solvent comprises methanol.

[0045] In some embodiments, the method further comprises heating the mixture to a temperature of about 70°C to about 90°C.

[0046] In some embodiments, the method further comprises heating the mixture to a temperature of about 50°C to about 70°C.

[0047] In some embodiments, the method further comprises cooling the mixture to room temperature.

[0048] The present application also provides a crystalline form of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionamide, which is prepared according to the method provided herein. In some embodiments, the crystalline form is Form IA as described herein. In some embodiments, the crystalline form is Form IIA as described herein. In some embodiments, the crystalline form is Form IIIA as described herein.

[0049] In some embodiments, the present application provides a method for preparing a hydrobromide salt of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionamide. In some embodiments, the method comprises dissolving a compound of formula A in a solvent to form a mixture, and adding hydrobromic acid to the mixture.

[0050] In some embodiments of the method of preparing a hydrobromide salt, the solvent comprises methanol.

[0051] In some embodiments of the method of preparing a hydrobromide salt, the hydrobromic acid is added to the mixture as an aqueous hydrobromic acid solution.

[0052] In some embodiments of the method of preparing the hydrobromide salt, excess hydrobromic acid is added to the mixture based on 1 equivalent of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0053] In some embodiments of the method for preparing the hydrobromide salt, about 1.1 to about 1.5 equivalents of hydrobromic acid are added to the mixture based on 1 equivalent of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0054] In some embodiments of the method of preparing the hydrobromide salt, the method further comprises substantially isolating the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide salt.

[0055] In some embodiments of the method of preparing the hydrobromide salt, the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide salt is isolated in a crystalline form.

[0056] In some embodiments of the method for preparing the hydrobromide salt, the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide salt is isolated as a methanol solvate crystalline form.

[0057] In some embodiments, the application also provides a hydrobromide salt of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, which is prepared according to the method provided herein. In some embodiments, the hydrobromide salt is crystalline. In some embodiments, the hydrobromide salt is a solvated crystalline form. In some embodiments, the hydrobromide salt is a methanol solvate crystalline form.

[0058] In some embodiments, the crystalline forms of the present invention are substantially isolated. "Substantially isolated" means that a particular crystalline form of a compound of Formula A is at least partially separated from impurities. For example, in some embodiments, the crystalline forms of the present invention contain less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Impurities generally include any material other than the substantially isolated crystalline form, including, for example, other crystalline forms and other materials.

[0059] In some embodiments, the crystalline form of the compound of Formula A is substantially free of other crystalline forms. The phrase "substantially free of other crystalline forms" means that a particular crystalline form of the compound of Formula A comprises more than about 80%, more than about 90%, more than about 95%, more than about 98%, more than about 99%, or more than about 99.5% by weight of the particular crystalline form.

[0060] Crystalline Form IA of the compound of formula A

[0061] In some embodiments, the crystalline form of the compound of Formula A is Form IA. In some embodiments, the crystalline Form IA of the compound of Formula A is anhydrous and non-solvated. The preparation of the Form IA compound of the compound of Formula A is described in Example 1. The crystalline Form IA of the compound of Formula A can generally be prepared as described in Example 2.

[0062] Crystalline Form IA of the compound of Formula A can be identified by unique signatures associated with, for example, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, the crystalline Form IA of the compound of Formula A is substantially as follows Figure 1 The XRPD pattern shown is characteristic. The peaks of the XRPD pattern are listed in Table 1.

[0063] In some embodiments, the crystalline Form IA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 8.6°±0.2°. In some embodiments, the crystalline Form IA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 9.5°±0.2°. In some embodiments, the crystalline Form IA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 10.3°±0.2°. In some embodiments, the crystalline Form IA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 14.9°±0.2°.

[0064] In some embodiments, crystalline Form IA of the compound of Formula A has an XRPD pattern comprising the following peaks, expressed in 2θ: 8.6°±0.2°; 9.5°±0.2°; 10.3°±0.2°; 13.0°±0.2°; 13.6°±0.2°; 14.2°±0.2°; and 14.9°±0.2°. In some embodiments, crystalline Form IA of the compound of Formula A has an XRPD pattern comprising the following peaks, expressed in 2θ: 8.6°±0.2°; 9.5°±0.2°; 10.3°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 17.8°±0.2°; 19.0°±0.2°; 19.2°±0.2°; 20.1°±0.2°; 20.6°±0.2°; 21.2°±0.2°; 22.2°±0.2°; 24.0°±0.2°; 26.8°±0.2°; and 28.7°±0.2°. In some embodiments, crystalline Form IA of the compound of Formula A has an XRPD pattern comprising the following peaks expressed in 2θ: 10.3°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 19.2°±0.2°; and 24.0°±0.2°.

[0065] In some embodiments, crystalline Form IA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 8.6°±0.2°; 9.5°±0.2°; 10.3°±0.2°; 13.0°±0.2°; 13.6°±0.2°; 14.2°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 19.2°±0.2°; 20.6°±0.2°; 24.0°±0.2°; and 28.7°±0.2°. In some embodiments, crystalline Form IA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 8.6°±0.2°; 9.5°±0.2°; 10.3°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 17.8°±0.2°; 19.0°±0.2°; 19.2°±0.2°; 20.1°±0.2°; 20.6°±0.2°; 21.2°±0.2°; 22.2°±0.2°; 24.0°±0.2°; 26.8°±0.2°; and 28.7°±0.2°. In some embodiments, crystalline Form IA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 9.5°±0.2°; 10.3°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 19.2°±0.2°; 20.6°±0.2°; 24.0°±0.2°; and 28.7°±0.2°.

[0066] In some embodiments, Form IA of the compound of Formula A is characterized by a DSC thermogram comprising an endothermic peak having a maximum at about 193° C. In some embodiments, the crystalline Form IA of the compound of Formula A has substantially Figure 2 DSC thermogram shown.

[0067] In some embodiments, the crystalline Form IA of the compound of Formula A has substantially Figure 3 TGA trace shown.

[0068] Crystalline Form IIA of the compound of formula A

[0069] In some embodiments, the crystalline form of the compound of Formula A is Form IIA. In some embodiments, the crystalline Form IIA of the compound of Formula A is anhydrous and non-solvated. This crystalline form can generally be prepared as described in Example 3.

[0070] Crystalline Form IIA of the compound of Formula A can be identified by unique signatures associated with, for example, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, the crystalline Form IIA of the compound of Formula A is substantially as follows Figure 4 The XRPD pattern shown is characteristic. The peaks of the XRPD pattern are listed in Table 2.

[0071] In some embodiments, the crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 9.1±0.2°. In some embodiments, the crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 11.1°±0.2°. In some embodiments, the crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 21.9°±0.2°. In some embodiments, the crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 12.6°±0.2°. In some embodiments, the crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 13.6°±0.2°. In some embodiments, the crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 18.0°±0.2°. In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 19.0°±0.2°.

[0072] In some embodiments, crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising the following peaks expressed in 2θ: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; and 13.5°±0.2°. In some embodiments, crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising the following peaks, expressed in 2θ: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; 13.5°±0.2°; 16.1°±0.2°; 16.9°±0.2°; 18.0°±0.2°; 18.4°±0.2°; 19.0°±0.2°; 19.7°±0.2°; 20.1°±0.2°; 20.5°±0.2°; 21.9°±0.2°; 23.7°±0.2°; 23.8°±0.2°; 25.1°±0.2°; 25.3°±0.2°; 25.8°±0.2°; and 27.3°±0.2°. In some embodiments, crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising the following peaks expressed in 2θ: 12.6°±0.2°; 18.0°±0.2°; 19.0°±0.2°; and 21.9°±0.2°.

[0073] In some embodiments, the crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; 13.5°±0.2°; 18.0°±0.2°; 19.0°±0.2°; 20.5°±0.2°; and 21.9°±0.2°. In some embodiments, the crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; 13.5°±0.2°; 16.1°±0.2°; 16.9°±0.2°; 18.0°±0.2°; 19.0°±0.2°; 20.5°±0.2°; and 21.9°±0.2°. In some embodiments, the crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising two or more, three or more, or four or more of the following peaks, expressed in 2θ: 12.6°±0.2°; 18.0°±0.2°; 19.0°±0.2°; 20.5°±0.2°; and 21.9°±0.2°.

[0074] In some embodiments, Form IIA of the compound of Formula A is characterized by a DSC thermogram comprising an endothermic peak with a maximum at about 180°C. In some embodiments, the crystalline Form IIA of the compound of Formula A has substantially Figure 5 DSC thermogram shown.

[0075] In some embodiments, the crystalline Form IIA of the compound of Formula A has substantially Figure 6 TGA trace shown.

[0076] Crystalline Form IIIA of the compound of formula A

[0077] In some embodiments, the crystalline form of the compound of Formula A is Form IIIA. In some embodiments, the crystalline Form IIIA of the compound of Formula A is anhydrous and non-solvated. This crystalline form can generally be prepared as described in Example 4.

[0078] Crystalline Form IIIA of the compound of Formula A can be identified by unique signatures associated with, for example, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, the crystalline Form IIIA of the compound of Formula A is substantially as follows Figure 7 The XRPD pattern shown is characteristic. Table 3 lists the peaks of the XRPD pattern.

[0079] In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 8.1°±0.2°. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 10.6°±0.2°. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 13.5°±0.2°. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 14.2°±0.2°. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern comprising a peak expressed in 2θ at 20.3°±0.2°.

[0080] In some embodiments, the crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising the following peaks, expressed in 2θ: 8.1°±0.2°; 10.6°±0.2°; 13.5°±0.2°; and 14.2°±0.2°. In some embodiments, the crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising the following peaks, expressed in 2θ: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; 16.4°±0.2°; 17.1°±0.2°; 17.9°±0.2°; 20.3°±0.2°; 20.8°±0.2°; 24.1°±0.2°; 24.6°±0.2°; 24.8°±0.2°; and 27.5°±0.2°. In some embodiments, crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising the following peaks expressed in 2θ: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; and 20.3°±0.2°.

[0081] In some embodiments, crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 8.1°±0.2°; 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; 16.4°±0.2°; 17.1°±0.2°; 17.9°±0.2°; 20.3°±0.2°; and 24.1°±0.2°. In some embodiments, crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; 16.4°±0.2°; 17.1°±0.2°; 17.9°±0.2°; 20.3°±0.2°; 20.8°±0.2°; 24.1°±0.2°; 24.6°±0.2°; 24.8°±0.2°; and 27.5°±0.2°. In some embodiments, crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; 16.4°±0.2°; 17.1°±0.2°; 17.9°±0.2°; 20.3°±0.2°; and 24.1°±0.2°.

[0082] In some embodiments, Form IIIA of the compound of Formula A is characterized by a DSC thermogram comprising an endothermic peak having a maximum at about 143°C. In some embodiments, the crystalline Form IIIA of the compound of Formula A has substantially Figure 8 DSC thermogram shown.

[0083] In some embodiments, the crystalline Form IIIA of the compound of Formula A has substantially Fig. 9 TGA trace shown.

[0084] The hydrobromide salt of formula A

[0085] In some embodiments, the present application provides a hydrobromide salt of a compound of formula A. In some embodiments, the hydrobromide salt of a compound of formula A has a 1:1 stoichiometric ratio of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionamide to hydrobromic acid. The hydrobromide salt form can generally be prepared as described in Example 11.

[0086] In some embodiments, the hydrobromide salt of the compound of formula A is crystalline. In some embodiments, the hydrobromide salt of the compound of formula A is a solvated crystalline form. In some embodiments, the hydrobromide salt of the compound of formula A is a methanol solvate crystalline form.

[0087] Crystalline form of the compound of formula B

[0088] The compound of formula B can be isolated in a variety of crystalline forms, including, for example, anhydrous and / or non-solvated crystalline forms. In some embodiments, the crystalline form of the compound of formula B is anhydrous. In some embodiments, the crystalline form of the compound of formula B is non-solvated. In some embodiments, the crystalline form of the compound of formula B is anhydrous and non-solvated. "Anhydrous" means that the crystalline form of the compound of formula B is substantially free of bound water in the crystal lattice structure, that is, the compound does not form a crystalline hydrate.

[0089] In some embodiments, the crystalline form provided herein can be prepared, for example, by a method comprising dissolving 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide in a solvent to form a mixture and crystallizing the compound from the mixture.

[0090] In some embodiments, the method further comprises heating the mixture to a temperature of about 70° C. to about 90° C. In some embodiments, the method further comprises cooling the mixture to room temperature.

[0091] In some embodiments, the solvent comprises isopropyl acetate. In some embodiments, the solvent further comprises heptane.

[0092] In some embodiments, the present application provides a crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, which is prepared according to the method provided herein. In some embodiments, the crystalline form is Form IB as described herein. In some embodiments, the crystalline form is Form IIB as described herein.

[0093] In some embodiments, the crystalline forms of the present invention are substantially isolated. "Substantially isolated" means at least partially separating a particular crystalline form of a compound of Formula B from impurities. For example, in some embodiments, the crystalline forms of the present invention contain less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Impurities generally include any material other than the substantially isolated crystalline form, including, for example, other crystalline forms and other materials.

[0094] In some embodiments, the crystalline form of the compound of Formula B is substantially free of other crystalline forms. The phrase "substantially free of other crystalline forms" means that a particular crystalline form of the compound of Formula B comprises greater than about 80%, greater than about 90%, greater than about 95%, greater than about 98%, greater than about 99%, or greater than about 99.5% by weight of the particular crystalline form.

[0095] Crystalline Form IB of the compound of formula B

[0096] In some embodiments, the crystalline form of the compound of Formula B is Form IB. In some embodiments, the crystalline Form IB of the compound of Formula B is anhydrous and non-solvated. The preparation of the compound of Formula B is described in Examples 5 and 6. Crystalline Form IB can generally be prepared as described in Example 7.

[0097] Crystalline Form IB of the compound of Formula B can be identified by unique signatures associated with, for example, X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the crystalline Form IB of the compound of Formula B is substantially as follows Fig.10 The XRPD pattern shown is characteristic. Table 4 lists the peaks of the XRPD pattern.

[0098] In some embodiments, the crystalline Form IB of the compound of Formula B is characterized by an XRPD pattern comprising a peak at 6.2°±0.2°, expressed in 2θ. In some embodiments, the crystalline Form IB of the compound of Formula B is characterized by an XRPD pattern comprising a peak at 15.6°±0.2°, expressed in 2θ. In some embodiments, the crystalline Form IB of the compound of Formula B is characterized by an XRPD pattern comprising a peak at 20.7°±0.2°, expressed in 2θ. In some embodiments, the crystalline Form IB of the compound of Formula B has an XRPD pattern comprising the following peaks, expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°. In some embodiments, crystalline Form IB of the compound of Formula B has an XRPD pattern comprising the following peaks, expressed in 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.4°±0.2°; 11.6°±0.2°; 12.0°±0.2°; 13.9°±0.2°; 14.4°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°. In some embodiments, crystalline Form IB of the compound of Formula B has an XRPD pattern comprising the following peaks, expressed in 2θ: 6.2°±0.2°; 12.0°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 19.3°±0.2°; 20.7°±0.2°; 23.2°±0.2°; and 27.1°±0.2°. In some embodiments, crystalline Form IB of the compound of Formula B has an XRPD pattern comprising the following peaks, expressed in 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.4°±0.2°; 11.6°±0.2°; 12.0°±0.2°; 13.9°±0.2°; and 14.4°±0.2°.

[0099] In some embodiments, crystalline Form IB of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.4°±0.2°; 11.6°±0.2°; 12.0°±0.2°; 13.9°±0.2°; 14.4°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°. In some embodiments, crystalline Form IB of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 6.2°±0.2°; 12.0°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 19.3°±0.2°; 20.7°±0.2°; 23.2°±0.2°; and 27.1°±0.2°. In some embodiments, the crystalline Form IB of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°. In some embodiments, the crystalline Form IB of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.4°±0.2°; 11.6°±0.2°; 12.0°±0.2°; 13.9°±0.2°; and 14.4°±0.2°.

[0100] In some embodiments, Form IB of the compound of Formula B is characterized by a DSC thermogram comprising an endothermic peak having a maximum at about 174°C. In some embodiments, the crystalline Form IB of the compound of Formula B has substantially Fig.11 DSC thermogram shown.

[0101] Crystalline Form IIB of the compound of formula B

[0102] In some embodiments, the crystalline form of the compound of Formula B is Form IIB. In some embodiments, the crystalline Form IIB of the compound of Formula B is anhydrous and non-solvated. This crystalline form can generally be prepared as described in Example 8.

[0103] Crystalline Form IIB of the compound of Formula B can be identified by unique signatures associated with, for example, X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the crystalline Form IIB of the compound of Formula B is substantially as follows Fig.12 The XRPD pattern shown is characteristic. Table 5 lists the peaks of the XRPD pattern.

[0104] In some embodiments, the crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern comprising a peak expressed in 2θ at 4.2°±0.2°. In some embodiments, the crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern comprising a peak expressed in 2θ at 7.4°±0.2°. In some embodiments, the crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern comprising a peak expressed in 2θ at 13.3°±0.2°. In some embodiments, the crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern comprising a peak expressed in 2θ at 20.1°±0.2°. In some embodiments, the crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern comprising a peak expressed in 2θ at 17.0°±0.2°. In some embodiments, the crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern comprising a peak expressed in 2θ at 18.8°±0.2°.

[0105] In some embodiments, crystalline Form IIB of the compound of Formula B has an XRPD pattern comprising the following peaks, expressed in 2θ: 4.3°±0.2°; 7.4°±0.2°; 13.3°±0.2°; and 15.3°±0.2°. In some embodiments, crystalline Form IIB of the compound of Formula B has an XRPD pattern comprising the following peaks, expressed in 2θ: 4.3°±0.2°; 7.4°±0.2°; 15.3°±0.2°; 17.0°±0.2°; 18.8°±0.2°; and 20.1°±0.2°. In some embodiments, crystalline Form IIB of the compound of Formula B has an XRPD pattern comprising the following peaks, expressed in 2θ: 4.3°±0.2°; 7.4°±0.2°; 13.3°±0.2°; 15.3°±0.2°; 15.5°±0.2°; 17.0°±0.2°; 17.2°±0.2°; 18.1°±0.2°; 18.8°±0.2°; 19.6°±0.2°; 20.1°±0.2°; 21.4°±0.2°; 23.5°±0.2°; 25.8°±0.2°; 26.2°±0.2°; and 27.3°±0.2°.

[0106] In some embodiments, crystalline Form IIB of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 4.3°±0.2°; 7.4°±0.2°; 13.3°±0.2°; 15.3°±0.2°; 15.5°±0.2°; 17.0°±0.2°; 17.2°±0.2°; 18.1°±0.2°; 18.8°±0.2°; 19.6°±0.2°; 20.1°±0.2°; 21.4°±0.2°; 23.5°±0.2°; 25.8°±0.2°; 26.2°±0.2°; and 27.3°±0.2°. In some embodiments, crystalline Form IIB of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 4.3°±0.2°; 7.4°±0.2°; 13.3°±0.2°; 15.3°±0.2°; 15.5°±0.2°; 17.0°±0.2°; 17.2°±0.2°; 18.8°±0.2°; and 20.1°±0.2°.

[0107] In some embodiments, Form IIB of the compound of Formula B is characterized by a DSC thermogram comprising an endothermic peak having a maximum at about 165°C. In some embodiments, the crystalline Form IIB of the compound of Formula B has substantially Fig.13 DSC thermogram shown.

[0108] Crystalline form of the compound of formula C

[0109] The compound of formula C can be isolated in a variety of crystalline forms, including, for example, anhydrous and / or non-solvated crystalline forms. In some embodiments, the crystalline form of the compound of formula C is anhydrous. In some embodiments, the crystalline form of the compound of formula C is non-solvated. In some embodiments, the crystalline form of the compound of formula C is anhydrous and non-solvated. "Anhydrous" means that the crystalline form of the compound of formula C is substantially free of bound water in the crystal lattice structure, that is, the compound does not form a crystalline hydrate.

[0110] In some embodiments, the present application provides a method for preparing the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 In some embodiments, the method comprises 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide is dissolved in a solvent to form a mixture and the compound is crystallized from the mixture.

[0111] In some embodiments, the method further comprises heating the mixture to a temperature of about 70° C. to about 90° C. In some embodiments, the method further comprises cooling the mixture to room temperature.

[0112] In some embodiments, the solvent comprises isopropyl acetate. In some embodiments, the solvent further comprises heptane.

[0113] The present application also provides a compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide crystalline form prepared according to the methods provided herein. In some embodiments, the crystalline form is Form IC as described herein.

[0114] In some embodiments, the crystalline forms of the present invention are substantially isolated. "Substantially isolated" means that a particular crystalline form of a compound of Formula C is at least partially separated from impurities. For example, in some embodiments, the crystalline forms of the present invention contain less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Impurities generally include any material other than the substantially isolated crystalline form, including, for example, other crystalline forms and other materials.

[0115] In some embodiments, the crystalline form of the compound of Formula C is substantially free of other crystalline forms. The phrase "substantially free of other crystalline forms" means that a particular crystalline form of the compound of Formula C comprises greater than about 80%, greater than about 90%, greater than about 95%, greater than about 98%, greater than about 99%, or greater than about 99.5% by weight of the particular crystalline form.

[0116] Crystalline Form IC of the compound of formula C

[0117] In some embodiments, the crystalline form of the compound of Formula C is crystalline Form IC. In some embodiments, the crystalline Form IC of the compound of Formula C is anhydrous and non-solvated. Example 9 describes the preparation of the compound of Formula C. The crystalline Form IC of the compound of Formula C can generally be prepared as described in Example 10.

[0118] Crystalline Form IC of the compound of Formula C can be identified by unique signatures associated with, for example, X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the crystalline Form IC of the compound of Formula C is substantially as follows Fig.14 The XRPD pattern shown is characteristic. Table 6 lists the peaks of the XRPD pattern.

[0119] In some embodiments, crystalline Form IC of the compound of Formula C is characterized by an XRPD pattern comprising a peak expressed in 2θ at 6.2°±0.2°. In some embodiments, crystalline Form IC of the compound of Formula C is characterized by an XRPD pattern comprising a peak expressed in 2θ at 11.9°±0.2°. In some embodiments, crystalline Form IC of the compound of Formula C is characterized by an XRPD pattern comprising a peak expressed in 2θ at 16.7°±0.2°. In some embodiments, crystalline Form IC of the compound of Formula C has an XRPD pattern comprising the following peaks, expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 18.8°±0.2°; 19.9°±0.2°; 20.7°±0.2°; 21.2°±0.2°; 22.3°±0.2°; 23.2°±0.2°; and 27.0°±0.2°. In some embodiments, crystalline Form IC of the compound of Formula C has an XRPD pattern comprising the following peaks, expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; and 21.2°±0.2°. In some embodiments, crystalline Form IC of the compound of Formula C has an XRPD pattern comprising the following peaks expressed in 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.3°±0.2°; 11.9°±0.2°; and 12.5°±0.2°.

[0120] In some embodiments, crystalline Form IC of the compound of Formula C has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 18.8°±0.2°; 19.9°±0.2°; 20.7°±0.2°; 21.2°±0.2°; 22.3°±0.2°; 23.2°±0.2°; and 27.0°±0.2°. In some embodiments, crystalline Form IC of the compound of Formula C has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.3°±0.2°; 11.9°±0.2°; 12.5°±0.2°; 13.8°±0.2°; 14.4°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 21.2°±0.2°. In some embodiments, crystalline Form IC of the compound of Formula C has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 21.2°±0.2°.

[0121] In some embodiments, Form IC of the compound of Formula C is characterized by a DSC thermogram comprising an endothermic peak with a maximum at about 179° C. In some embodiments, the crystalline Form IC of the compound of Formula C has substantially Fig.15 DSC thermogram shown.

[0122] How to use

[0123] The compounds of the invention described herein (e.g., salts and crystalline forms) inhibit the activity of PI3Kγ kinase. Therefore, the salts and crystalline forms of the invention described herein can be used for methods of inhibiting the kinase by contacting PI3Kγ kinase. In some embodiments, the salts and crystalline forms of the invention can be used for methods of inhibiting the PI3Kγ activity of individuals / patients that need to be inhibited by administering an effective amount of the salts or crystalline forms described herein. In some embodiments, regulation is inhibition. In some embodiments, contact occurs in vivo. In some embodiments, contact occurs in vitro. Advantageously, the crystalline forms described herein show better efficacy and favorable safety and toxicity profiles in animal studies.

[0124] In some embodiments, PI3Kγ includes a mutation. The mutation can be a substitution of one amino acid for another amino acid, or a deletion of one or more amino acids. In such embodiments, the mutation can be present in the kinase domain of PI3Kγ.

[0125] In some embodiments, the salt or crystalline form also inhibits PI3Kδ.

[0126] The salts and crystalline forms described herein can be selective. "Selectivity" means that the salt or crystalline form binds or inhibits PI3Kγ with greater affinity or efficacy than at least one other kinase, respectively. In some embodiments, the salts and crystalline forms of the present invention are selective inhibitors of selectivity to PI3Kγ that exceeds the selectivity to PI3Kδ, PI3Kα and PI3Kβ. In some embodiments, the salts and crystalline forms of the present invention are selective inhibitors of selectivity to PI3Kγ that exceeds the selectivity to PI3Kα and PI3Kβ. In some embodiments, the selectivity can be at least about 2 times, 3 times, 5 times, 10 times or 20 times that of PI3Kδ, as measured by the assay described herein. In some embodiments, the selectivity of each enzyme can be tested at 2 μM ATP concentration. In some embodiments, the selectivity of the salts and crystalline forms of the present invention can be determined by cell assays associated with specific PI3K kinase activity.

[0127] Another aspect of the present invention relates to a method for treating a kinase PI3Kγ-related disease or disorder in an individual (e.g., a patient) in need of treatment by administering a therapeutically effective amount or dose of one or more salts or crystalline forms of the present invention or a pharmaceutical composition thereof. PI3Kγ-related diseases or disorders may include any disease, disorder, or condition directly or indirectly related to the expression or activity of PI3Kγ, including overexpression and / or abnormal activity levels.

[0128] In some embodiments, the disease or disorder is an autoimmune disease or disorder, cancer, cardiovascular disease, or a neurodegenerative disease.

[0129] In some embodiments, the disease or disorder is lung cancer (e.g., non-small cell lung cancer), melanoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, colon cancer, endometrial cancer, bladder cancer, skin cancer, uterine cancer, kidney cancer, gastric cancer, or sarcoma. In some embodiments, the sarcoma is Askin's tumor, botryoid sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft part sarcoma, angiosarcoma, cystic sarcoma phyllodes, dermatofibrosarcoma protuberans, desmoid tumor, desmoplastic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, hemangioendothelioma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma, or undifferentiated pleomorphic sarcoma.

[0130] In some embodiments, the disease or condition is mesothelioma or adenocarcinoma. In some embodiments, the disease or condition is mesothelioma. In some embodiments, the disease or condition is adenocarcinoma.

[0131] In some embodiments, the disease or disorder is acute myeloid leukemia (e.g., acute monocytic leukemia), small lymphocytic lymphoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma, T-cell acute lymphoblastic leukemia (T-ALL), cutaneous T-cell lymphoma, large granular lymphocytic leukemia, mature (peripheral) T-cell neoplasm (PTCL), anaplastic large cell lymphoma (ALCL), or lymphoblastic lymphoma. In some embodiments, the mature (peripheral) t-cell neoplasm (PTCL) is T-cell prolymphocytic leukemia, T-cell granular lymphocytic leukemia, aggressive NK-cell leukemia, mycosis fungoides / Sezary syndrome, anaplastic large cell lymphoma (T-cell type), enteropathy-type T-cell lymphoma, adult T-cell leukemia / lymphoma or angioimmunoblastic T-cell lymphoma. In some embodiments, anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL.

[0132] In some embodiments, the disease or disorder is Burkitt's lymphoma, acute myeloblastic leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanthoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenstrom's macroglobulinemia, macroglobulinemia), prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, latent myeloma (also called asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), or diffuse large B-cell lymphoma.

[0133] MDSC (myeloid-derived suppressor cells) are a heterologous immune cell population from the bone marrow lineage (a family of cells derived from bone marrow stem cells). Due to changes in hematopoiesis, MDSC is strongly amplified in pathological situations such as chronic infection and cancer. The difference between MDSC and other bone marrow cell types is that they have strong immunosuppressive activity rather than immunostimulatory properties. Similar to other bone marrow cells, MDSC interacts with other immune cell types, including T cells, dendritic cells, macrophages, and natural killer cells, to regulate their functions. In some embodiments, the compounds described herein, etc. can be used in methods related to cancerous tissues (e.g., tumors) with high infiltration of MDSC, including solid tumors infiltrated with macrophages and / or MDSC with high basal levels.

[0134] In some embodiments, the disease or disorder is Burkitt's lymphoma, acute myeloblastic leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenstrom's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, latent myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), or diffuse large B-cell lymphoma.

[0135] In some embodiments, the non-Hodgkin's lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, indolent NHL (iNHL), or aggressive NHL (aNHL).

[0136] In some embodiments, the diffuse large B-cell lymphoma is activated B-cell-like (ABC) diffuse large B-cell lymphoma or germinal center B-cell (GCB) diffuse large B-cell lymphoma.

[0137] In some embodiments, the Burkitt's lymphoma is endemic Burkitt's lymphoma, sporadic Burkitt's lymphoma, or Burkitt's lymphoma-like.

[0138] In some embodiments, the disease or disorder is rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, asthma, allergic reactions (e.g., allergic rhinitis), pancreatitis, psoriasis, allergies, glomerulonephritis, inflammatory bowel disease (e.g., Crohn's disease and ulcerative colitis), thrombosis, meningitis, encephalitis, diabetic retinopathy, benign prostatic hypertrophy, myasthenia gravis, Sjögren's syndrome ( syndrome), osteoarthritis, restenosis, or atherosclerosis.

[0139] In some embodiments, the disease or disorder is cardiac hypertrophy, cardiomyocyte dysfunction, acute coronary syndrome, chronic obstructive pulmonary disease (COPD), chronic bronchitis, elevated blood pressure, ischemia, ischemia-reperfusion, vasoconstriction, anemia (e.g., hemolytic anemia, aplastic anemia, or pure red cell anemia), bacterial infection, viral infection, transplant rejection, kidney disease, anaphylactic shock fibrosis, skeletal muscle atrophy, skeletal muscle hypertrophy, angiogenesis, sepsis, graft-versus-host disease, allogeneic or xenogeneic transplantation, glomerulosclerosis, progressive renal fibrosis, idiopathic thrombocytopenic purpura (ITP), idiopathic pulmonary fibrosis, autoimmune hemolytic anemia, vasculitis, lupus nephritis, pemphigus, or membranous nephropathy.

[0140] In some embodiments, the disease or disorder is cardiac hypertrophy, cardiomyocyte dysfunction, chronic obstructive pulmonary disease (COPD), increased blood pressure, local ischemia, local ischemia-reperfusion, vasoconstriction, anemia (e.g., hemolytic anemia, aplastic anemia, or pure red cell anemia), bacterial infection, viral infection, transplant rejection, kidney disease, anaphylactic shock fibrosis, skeletal muscle atrophy, skeletal muscle hypertrophy, angiogenesis, sepsis, transplant rejection, glomerulosclerosis, progressive renal fibrosis, idiopathic thrombocytopenic purpura (ITP), autoimmune hemolytic anemia, vasculitis, systemic lupus erythematosus, lupus nephritis, pemphigus, or membranous nephropathy.

[0141] In some embodiments, the disease or condition is Alzheimer's disease, central nervous system trauma, or stroke.

[0142] In some embodiments, the idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP.

[0143] In some embodiments, the vasculitis is Behcet's disease ( disease, Cogan's syndrome, giant cell arteritis, polymyalgia rheumatica (PMR), Takayasu's arteritis, Buerger's disease (thromboangiitis obliterans), central nervous system vasculitis, Kawasaki disease, polyarteritis nodosa, Churg-Strauss syndrome, mixed cryoglobulinemic vasculitis (primary or HCV-induced), Henoch-Schwann purpura (Henoch-Schwann purpura), purpura) (HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener's granulomatosis, or antineutrophil cytoplasmic antibody-associated (ANCA) systemic vasculitis (AASV).

[0144] The present invention also provides a salt or crystalline form described herein for use in any of the methods described herein.

[0145] The present invention also provides the use of a crystalline form described herein, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for use in any of the methods described herein.

[0146] As used herein, the term "contacting" refers to bringing the indicated moieties together in an in vitro system or an in vivo system. For example, "contacting" PI3K with a salt or crystalline form of the invention includes administering a crystalline form of the invention to an individual or patient (such as a human) having PI3K, as well as, for example, introducing a salt or crystalline form of the invention into a sample containing cells or purified preparations containing PI3K.

[0147] It is believed that any of the salts and crystalline forms provided herein or embodiments thereof may have a satisfactory pharmacological profile and promising biopharmaceutical properties, such as toxicological profile, metabolic and pharmacokinetic properties, solubility and permeability. It will be appreciated that the determination of appropriate biopharmaceutical properties is within the knowledge of those skilled in the art, such as determination of cytotoxicity in cells or inhibition of certain targets or pathways to determine potential toxicity.

[0148] As used herein, the terms "subject" or "patient," which are used interchangeably, refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates, and most preferably humans.

[0149] As used herein, the phrase "therapeutically effective amount" refers to that amount of an active salt, crystalline form, or pharmaceutical agent that elicits the biological or medical response that is being sought by a researcher, veterinarian, medical physician, or other clinician in a tissue, system, animal, individual, or human.

[0150] As used herein, the terms "treating" or "treatment" may refer to one or more of the following: (1) inhibiting a disease, e.g., inhibiting a disease, disorder or condition in an individual who is experiencing or displaying the pathology or symptoms of the disease, disorder or condition (i.e., arresting further development of the pathology and / or symptomology); and (2) ameliorating a disease, e.g., ameliorating a disease, disorder or condition in an individual who is experiencing or displaying the pathology or symptoms of the disease, disorder or condition (i.e., reversing the pathology and / or symptomology), such as reducing the severity of the disease.

[0151] In some embodiments, the salts and crystalline forms of the invention are useful for preventing or reducing the risk of developing any of the diseases mentioned herein; for example, preventing or reducing the risk of developing a disease, disorder, or condition in an individual who may be susceptible to the disease, disorder, or condition but does not yet experience or display the pathology or symptomology of the disease.

[0152] Combination therapy

[0153] I. Immune Checkpoint Therapy

[0154] In some embodiments, the PI3Kγ inhibitor provided herein can be used in combination with one or more immune checkpoint inhibitors for the treatment of cancer as described herein. In one embodiment, it can be used to treat melanoma in combination with one or more immune checkpoint inhibitors as described herein. The salt and crystalline form of the present invention can be used in combination with one or more immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors for immune checkpoint molecules such as CD20, CD28, CD40, CD122, CD96, CD73, CD47, GITR, CSF1R, JAK, PI3Kδ, PI3Kγ, TAM, arginase, HPK1, CD137 (also known as 4-1BB), ICOS, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA, TIGIT, PD-1, PD-L1 and PD-L2. In some embodiments, immune checkpoint molecules are selected from the stimulatory checkpoint molecules of CD27, CD28, CD40, ICOS, OX40, GITR and CD137. In some embodiments, the immune checkpoint molecule is selected from the inhibitory checkpoint molecules of A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, TIGIT and VISTA. In some embodiments, the salts and crystalline forms of the present invention provided herein can be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors and TGFRβ inhibitors.

[0155] In some embodiments, the PI3Kγ inhibitors provided herein can be used in combination with one or more agonists of immune checkpoint molecules, such as OX40, CD27, OX40, GITR, and CD137 (also known as 4-1BB).

[0156] In some embodiments, the immune checkpoint molecule inhibitor is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0157] In some embodiments, the immune checkpoint molecule inhibitor is a PD-1 inhibitor, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), durvalumab , pidilizumab, SHR-1210, PDR001, MGA012, PDR001, AB122 or AMP-224. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibody is pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012. In some embodiments, the anti-PD1 antibody is SHR-1210. Other anticancer agents include antibody therapeutics such as 4-1BB (e.g., urelumab, utomilumab).

[0158] In some embodiments, the immune checkpoint molecule inhibitor is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

[0159] In some embodiments, the immune checkpoint molecule inhibitor is an inhibitor of PD-1 and PD-L1, such as an anti-PD-1 / PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1 / PD-L1 is MCLA-136.

[0160] In some embodiments, the inhibitor is MCLA-145.

[0161] In some embodiments, the immune checkpoint molecule inhibitor is a CTLA-4 inhibitor, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884 or CP-675,206.

[0162] In some embodiments, the immune checkpoint molecule inhibitor is a LAG3 inhibitor, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, or INCAGN2385.

[0163] In some embodiments, the immune checkpoint molecule inhibitor is a TIM3 inhibitor, such as an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453 or TSR-022.

[0164] In some embodiments, the immune checkpoint molecule inhibitor is a GITR inhibitor, such as an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323 or MEDI1873.

[0165] In some embodiments, the immune checkpoint molecule inhibitor is an OX40 agonist, such as an OX40 agonist antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562, MOXR-0916, PF-04518600, GSK3174998, or BMS-986178. In some embodiments, the OX40L fusion protein is MEDI6383.

[0166] In some embodiments, the immune checkpoint molecule inhibitor is a CD20 inhibitor, such as an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

[0167] The salts and crystalline forms of the present invention can be used in combination with bispecific antibodies. In some embodiments, one of the domains of the bispecific antibody targets PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3 or TGFβ receptor.

[0168] In some embodiments, the PI3Kγ inhibitors provided herein can be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitor is an inhibitor of IDO1, TDO or arginase. Examples of IDO1 inhibitors include epacadostat, NLG919, BMS-986205, PF-06840003, IOM2983, RG-70099 and LY338196.

[0169] As provided throughout, other compounds, inhibitors, agents, etc. can be combined with the compounds of the present invention in a single or sequential dosage form, or they can be administered simultaneously or sequentially as separate dosage forms.

[0170] II. Cancer Therapy

[0171] Cancer cell growth and survival can be affected by multiple signaling pathways. Therefore, combining different enzyme / protein / receptor inhibitors that exhibit different preferences for targets whose activities are regulated by them can be used to treat such disorders. Targeting more than one signaling pathway (or more than one biomolecule involved in a given signaling pathway) can reduce the likelihood of developing drug resistance in a cell population and / or reduce the toxicity of the treatment.

[0172] The salts and crystalline forms of the present invention can be used in combination with one or more other enzyme / protein / receptor inhibitors or one or more therapies to treat a variety of diseases, such as cancer. Examples of diseases and indications that can be treated with combination therapy include those described herein. Examples of cancer include solid tumors and liquid tumors, such as blood cancers.

[0173] One or more other agents, such as chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressants, immuno-oncology agents, metabolic enzyme inhibitors, chemokine receptor inhibitors and phosphatase inhibitors, and targeted therapies, such as Bcr-Abl, Flt-3, EGFR, HER2, JAK, c-MET, VEGFR, PDGFR, c-Kit, IGF-1R, RAF and FAK kinase inhibitors, such as those described in WO 2006 / 056399. Other agents such as therapeutic antibodies can be used in combination with the salts and crystalline forms of the present invention to treat PI3K-related diseases, disorders or conditions. The one or more other agents can be administered to the patient simultaneously or sequentially.

[0174] For example, the salts and crystalline forms as disclosed herein can be used in combination with one or more inhibitors of the following kinases for the treatment of cancers and other diseases or disorders described herein: Akt1, Akt2, Akt3, TGF-βR, PKA, PKG, PKC, CaM kinase, phosphatase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IGF-1R, IR-R, PDGFαR, PDGFβR, CSFIR, KIT, FLK -II, KDR / FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, Ron, Sea, TRKA, TRKB, TRKC, FLT3, VEGFR / Fl t2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK and B-Raf. Non-limiting examples of inhibitors that can be combined with the salts and crystalline forms of the invention for the treatment of cancers and other diseases and conditions described herein include FGFR inhibitors (FGFR1, FGFR2, FGFR3 or FGFR4, such as INCB54828, INCB62079 and INCB63904), JAK inhibitors (JAK1 and / or JAK2, such as ruxolitinib, baricitinib or INCB39110), IDO inhibitors (such as epacadostat, NLG919 or BMS-986205), LSD inhibitors (such as seratinib, ... 1 inhibitors (e.g., INCB59872 and INCB60003), TDO inhibitors, PI3Kδ inhibitors (e.g., INCB50797 and INCB50465), Pim inhibitors, CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl and Mer), histone deacetylase inhibitors (HDAC) such as HDAC8 inhibitors, angiogenesis inhibitors, interleukin receptor inhibitors, bromodomain and extra-terminal family member inhibitors (e.g., bromodomain inhibitors or BET inhibitors, such as INCB54329 and INCB57643), and adenosine receptor antagonists, or a combination thereof.

[0175] In some embodiments, the salts and crystalline forms described herein are administered with a PI3Kδ inhibitor. In some embodiments, the salts and crystalline forms described herein are administered with a JAK inhibitor. In some embodiments, the salts and crystalline forms described herein are administered with a JAK1 or JAK2 inhibitor (e.g., baricitinib or ruxolitinib). In some embodiments, the salts and crystalline forms described herein are administered with a JAK1 inhibitor. In some embodiments, the salts and crystalline forms described herein are administered with a JAK1 inhibitor that has selectivity for JAK1 over that for JAK2.

[0176] Examples of antibodies for use in combination therapy include, but are not limited to, Trastuzumab (e.g., anti-HER2), Ranibizumab (e.g., anti-VEGF-A), Bevacizumab (trade name Avastin, e.g., anti-VEGF), Panitumumab (e.g., anti-EGFR), Cetuximab (e.g., anti-EGFR), Rituxan (anti-CD20), and antibodies to c-MET.

[0177] One or more of the following agents may be used in combination with the salts and crystalline forms of the invention and are presented in a non-limiting list: cytostatics, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methoxtrexate, temozolomide, cyclophosphamide, SCH 66336, R115777, L778,123, BMS 214662, IRESSA TM (gefitinib), TARCEVA TM(erlotinib), antibodies against EGFR, intron, ara-C, adriamicin, cytoxan, gemcitabine, uracil mustard, nitrogen mustard, ifosfamide, melphalan, nitrogen mustard phenylbutyric acid, pipobroman, triethylene melamine, triethylene thiophosphamide, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, oxaliplatin, leucovirin, ELOXATIN TM(oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide, 17α-ethinyl estradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate), testolactone, megestrolacetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, Flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, navelbene, anastrazole, letrazole, capecitabine, reloxafine, droloxafine, hexamethylmelamine, acrivastine, HERCEPTIN TM (trastuzumab), BEXXAR TM(tositumomab), VELCADE TM (bortezomib), ZEVALIN TM (ibritumomab tiuxetan), TRISENOX TM (arsenic trioxide), XELODA TM (capecitabine), vinorelbine, porfimer, ERBITUX TM (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, letrozole, fulvestrant, exemestane, iforamide, rituximab, C225 (cetuximab), Campath (alemtuzumab), clofarabine, cladribine, aphidicolon, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triapine, didox, trimidox, amidox, 3-AP, and MDL-101,731.

[0178] The salt and crystalline form of the present invention can also be used in combination with other methods for treating cancer (e.g., by chemotherapy, irradiation therapy, tumor-targeted therapy, adjuvant therapy, immunotherapy or surgery). Examples of immunotherapy include cytokine therapy (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, adoptive T cell transfer, Toll-like receptor agonists, STING agonists, oncolytic virus therapy and immunomodulatory small molecules, including thalidomide or JAK1 / 2 inhibitors, etc. The salt and crystalline form can be administered in combination with one or more anticancer drugs, such as chemotherapeutic agents. Examples of chemotherapeutic agents include any of the following: abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, hexamethylmelamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezombi, bortezomib, intravenous busulfan, oral busulfan, calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, cloralbine, cyclophosphamide, cytarabine, dacarbazine, actinomycin, dalteparin sodium sodium), daunomycin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, fluuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate acetate), ibritumomab tiuxetan, idarubicin, ifofenamide, imatinib mesylatemesylate, interferon alfa-2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, nitrogen mustard, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mithramycin, mitomycin C, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab, olaparib, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, rucaparib, streptozocin, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, niraparib, veliparib, talazoparib, and zoledronate.

[0179] Other examples of chemotherapeutic agents include proteasome inhibitors (eg, bortezomib), thalidomide, revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

[0180] Examples of steroids include corticosteroids, such as dexamethasone or prednisone.

[0181] Examples of Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC TM), nilotinib, dasatinib, bosutinib and ponatinib and pharmaceutically acceptable salts. Other examples of suitable Bcr-Abl inhibitors include compounds of the class and species disclosed in U.S. Pat. No. 5,521,184, WO 04 / 005281 and U.S. Serial No. 60 / 578,491 and pharmaceutically acceptable salts thereof.

[0182] Examples of suitable Flt-3 inhibitors include midostaurin, lestaurtinib, linifanib, sunitinib, sunitinib, maleate, sorafenib, quizartinib, crenolanib, pacritinib, tandutinib, PLX3397 and ASP2215 and pharmaceutically acceptable salts thereof. Other examples of suitable Flt-3 inhibitors include compounds disclosed in WO 03 / 037347, WO 03 / 099771 and WO 04 / 046120 and pharmaceutically acceptable salts thereof.

[0183] Examples of suitable RAF inhibitors include dabrafenib, sorafenib and vemurafenib and pharmaceutically acceptable salts thereof.Other examples of suitable RAF inhibitors include compounds as disclosed in WO 00 / 09495 and WO 05 / 028444 and pharmaceutically acceptable salts thereof.

[0184] Examples of suitable FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520 and GSK2256098 and pharmaceutically acceptable salts thereof. Other examples of suitable FAK inhibitors include compounds disclosed in WO 04 / 080980, WO 04 / 056786, WO 03 / 024967, WO 01 / 064655, WO 00 / 053595 and WO 01 / 014402 and pharmaceutically acceptable salts thereof.

[0185] In some embodiments, the salts and crystalline forms of the invention can be used in combination with one or more other kinase inhibitors, including imatinib, particularly for treating patients who are resistant to imatinib or other kinase inhibitors.

[0186] In some embodiments, the salts and crystalline forms of the present invention can be used in combination with chemotherapeutic agents for the treatment of cancer, and compared with the response to a single chemotherapeutic agent, the therapeutic response can be improved without exacerbating its toxic effects. In some embodiments, the salts and crystalline forms of the present invention can be used in combination with chemotherapeutic agents provided herein. For example, other agents for the treatment of multiple myeloma may include, but are not limited to, melphalan, melphalan plus prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). Other additional agents for the treatment of multiple myeloma include Bcr-Abl, Flt-3, RAF, and FAK kinase inhibitors. In some embodiments, the agent is an alkylating agent, a proteasome inhibitor, a corticosteroid, or an immunomodulator. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulator is lenalidomide (LEN) or pomalidomide (POM).Additive or synergistic effects are desirable outcomes of combining the PI3K inhibitors of the invention with additional agents.

[0187] In some embodiments, the salts and crystalline forms of the invention may be used in combination with a JAK or PI3Kδ inhibitor.

[0188] The agents may be combined with the compounds of the invention in a single or sequential dosage form, or the agents may be administered simultaneously or sequentially as separate dosage forms.

[0189] The salts and crystalline forms of the invention can be used in combination with one or more other inhibitors or one or more therapies to treat infections. Examples of infections include viral infections, bacterial infections, fungal infections or parasitic infections.

[0190] In some embodiments, a corticosteroid such as dexamethasone is administered to a patient in combination with a compound of the invention, wherein the dexamethasone is administered intermittently as opposed to continuously.

[0191] The salts and crystalline forms of the invention as described herein can be combined with another immunogenic agent, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides and carbohydrate molecules), cells and cells transfected with genes encoding immunostimulatory cytokines. Non-limiting examples of tumor vaccines that can be used include peptides of melanoma antigens, such as gp100, MAGE antigens, Trp-2, MARTI and / or tyrosinase, or tumor cells transfected to express the cytokine GM-CSF.

[0192] The salts and crystalline forms of the invention as described herein can be used in combination with vaccination regimens to treat cancer. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins from viruses associated with human cancers, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). In some embodiments, the salts and crystalline forms of the invention can be used in combination with tumor-specific antigens, such as heat shock proteins isolated from tumor tissue itself. In some embodiments, the salts and crystalline forms can be combined with dendritic cell immunization to activate an effective anti-tumor response.

[0193] The salts and crystalline forms of the invention can be used in combination with bispecific macrocyclic peptides that target effector cells expressing Feα or Feγ receptors to tumor cells. The salts and crystalline forms of the invention can also be combined with macrocyclic peptides that activate host immune responsiveness.

[0194] In some other embodiments, the salts and crystalline forms of the present invention in combination with other therapeutic agents can be administered to a patient before, during and / or after a bone marrow transplant or stem cell transplant. The salts and crystalline forms of the present invention can be used in combination with a bone marrow transplant to treat a variety of hematopoietic tumors.

[0195] The salts and crystalline forms can be used in combination with vaccines to stimulate immune responses to pathogens, toxins and self-antigens. Examples of pathogens for which this treatment method may be particularly useful include pathogens for which there is currently no effective vaccine or for which traditional vaccines are not fully effective. These include, but are not limited to, HIV, hepatitis viruses (type A, B and C), influenza viruses, herpes viruses, flagellates, malaria, Leishmania, Staphylococcus aureus, Pseudomonas Aeruginosa.

[0196] Viruses that cause infections treatable by the methods of the invention include, but are not limited to, human papillomavirus, influenza virus, hepatitis A, B, C, or D virus, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory syndrome virus, ebola virus, measles virus, herpes virus (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein Barr virus), flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papilloma virus, molluscum virus, polio virus, rabies virus, JC virus, and arboviral encephalitis virus.

[0197] Pathogenic bacteria that cause infections treatable by the methods of the invention include, but are not limited to, chlamydia, rickettsial bacteria, mycobacteria, staphylococci, streptococci, pneumonococci, meningococci and conococci, klebsiella, proteus, serratia, pseudomonas, legionella, diphtheria, salmonella, bacilli, cholera, tetanus, botulism, anthrax, plague, leptospirosis, and Lyme's disease bacteria.

[0198] Pathogenic fungi causing infections treatable by the methods of the present invention include, but are not limited to, Candida (Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, etc.), Cryptococcus neoformans, Aspergillus (Aspergillus fumigatus, Aspergillus niger, etc.), Genus Mucorales (mucor, absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, and the like. immitis and Histoplasma capsulatum. Pathogenic parasites that cause infections treatable by the methods of the present invention include, but are not limited to, Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, and Nippostrongylus brasiliensis.

[0199] The methods of safely and effectively using most of these chemotherapeutics are known to those skilled in the art. In addition, their use is described in standard documents. For example, the use of many chemotherapeutics is described in "Physicians' Desk Reference" (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is incorporated herein by reference as if set forth in its entirety.

[0200] Drug preparations and dosage forms

[0201] When used as a medicament, the compounds of the present invention (e.g., salts and crystalline forms) can be administered in the form of pharmaceutical compositions. These compositions can be prepared in a manner well known in the pharmaceutical art and can be administered by a variety of routes, depending on whether local or systemic treatment is required and the area to be treated. Administration can be topical (including transdermal, epidermal, ocular and mucosal, including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular or injection or infusion; or intracranial (e.g., intrathecal) or intraventricular administration. Parenteral administration can be in the form of a single bolus dose, or can be, for example, by continuous infusion pumps. Pharmaceutical compositions and preparations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Traditional pharmaceutical carriers, aqueous solutions, powders or oily bases, thickeners, etc. may be necessary or desirable.

[0202] The present invention also includes pharmaceutical compositions, which contain a combination of the compounds of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient and one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the composition is suitable for topical administration. When manufacturing the composition of the present invention, the active ingredient is usually mixed with an excipient, diluted or encapsulated in a carrier such as a capsule, sachet, paper or other container by an excipient. When an excipient acts as a diluent, it can be a solid, semisolid or liquid material, acting as a medium, carrier or medium for the active ingredient. Therefore, the composition can be in the form of tablets, pills, powders, diamond-shaped tablets, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (in solid form or in a liquid medium), ointments containing, for example, up to 10% by weight of active compounds, soft gelatin capsules and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

[0203] When preparing a formulation, the active compound may be ground to provide an appropriate particle size before combining with the other ingredients. If the active compound is substantially insoluble, it may be ground to a particle size of less than 200 mesh. If the active compound is substantially soluble in water, the particle size may be adjusted by grinding to provide a substantially uniform distribution in the formulation, e.g., about 40 mesh.

[0204] The compounds of the invention can be ground using known grinding procedures, such as wet milling, to obtain a particle size suitable for tablet formation and for other formulation types. Fine powder (nanoparticulate) formulations of the compounds of the invention can be prepared by methods known in the art, for example, see International Application No. WO 2002 / 000196.

[0205] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose. The formulation may additionally include: lubricants, such as talc, magnesium stearate and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives, such as methyl benzoate and propyl hydroxybenzoate; sweeteners; and flavoring agents. The compositions of the present invention may be formulated so as to release the active ingredient quickly, continuously or delayed after administration to a patient using procedures known in the art.

[0206] The compositions may be formulated in unit dosage form, each dose containing from about 5 mg to about 1000 mg (1 g), more usually from about 100 mg to about 500 mg, of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unit dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect and in association with a suitable pharmaceutical excipient.

[0207] In some embodiments, the compositions of the present invention contain about 5 mg to about 50 mg of active ingredient. One of ordinary skill in the art will appreciate that this includes compositions containing about 5 mg to about 10 mg, about 10 mg to about 15 mg, about 15 mg to about 20 mg, about 20 mg to about 25 mg, about 25 mg to about 30 mg, about 30 mg to about 35 mg, about 35 mg to about 40 mg, about 40 mg to about 45 mg, or about 45 mg to about 50 mg of active ingredient.

[0208] In some embodiments, the compositions of the present invention contain about 50 to about 500 mg of active ingredient. Those skilled in the art will appreciate that this includes compositions containing about 50 mg to about 100 mg, about 100 mg to about 150 mg, about 150 mg to about 200 mg, about 200 mg to about 250 mg, about 250 mg to about 300 mg, about 350 mg to about 400 mg, or about 450 mg to about 500 mg of active ingredient.

[0209] In some embodiments, the compositions of the present invention contain from about 500 mg to about 1000 mg of active ingredient. One of ordinary skill in the art will appreciate that this includes compositions containing from about 500 mg to about 550 mg, from about 550 mg to about 600 mg, from about 600 mg to about 650 mg, from about 650 mg to about 700 mg, from about 700 mg to about 750 mg, from about 750 mg to about 800 mg, from about 800 mg to about 850 mg, from about 850 mg to about 900 mg, from about 900 mg to about 950 mg, or from about 950 mg to about 1000 mg of active ingredient.

[0210] Similar dosages may be used in the methods and uses of the invention with the compounds described herein.

[0211] The active compound can be effective within a wide dosage range and is generally administered in a therapeutically effective amount. However, it should be understood that the amount of compound actually administered will generally be determined by a physician based on the relevant circumstances, including the condition to be treated, the selected route of administration, the actual compound administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc.).

[0212] In order to prepare solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient to form a solid preformulated composition containing a homogeneous mixture of the compound of the invention. When these preformulated compositions are referred to as homogeneous, the active ingredient is generally evenly dispersed throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms, such as tablets, pills and capsules. This solid preformulation is then subdivided into the above-mentioned type of unit dosage forms containing, for example, about 0.1 mg to about 1000 mg of the active ingredient of the invention.

[0213] Tablets or pills of the present invention can be coated or otherwise compounded to provide a dosage form that can obtain the advantage of extended action time. For example, tablets or pills can include an inner dose and an outer dose component, the latter being in the form of an envelope on the former. The two components can be separated by an enteric coating layer, which is used to resist disintegration in the stomach and allow the inner component to enter the duodenum intact or delay release. A variety of materials can be used for such enteric coating layers or coatings, including many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol and cellulose acetate.

[0214] Liquid forms in which the compounds and compositions of the invention may be incorporated for oral or injection administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0215] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the composition is administered by oral or nasal respiratory route to achieve local or systemic effects. The composition may be atomized using an inert gas. The atomized solution may be breathed directly from the atomizing device, or the atomizing device may be connected to a mask, tent, or intermittent positive pressure breathing machine. The solution, suspension, or powder composition may be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0216] The topical preparation can contain one or more traditional carriers. In some embodiments, the ointment can contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white vaseline, etc. The carrier composition of the cream can be based on a combination of water with glycerol and one or more other components (such as glyceryl monostearate, PEG-glyceryl monostearate and cetyl stearyl alcohol). The gel can be formulated using a suitable combination of isopropyl alcohol and water with other components (such as glycerol, hydroxyethyl cellulose, etc.). In some embodiments, the topical preparation contains at least about 0.1wt%, at least about 0.25wt%, at least about 0.5wt%, at least about 1wt%, at least about 2wt% or at least about 5wt% of the compounds of the present invention. The topical preparation can be suitably packaged in, for example, a 100g tube, which is optionally associated with instructions for treating selected indications (such as psoriasis or other skin disorders).

[0217] The amount of compound or composition administered to a patient will vary depending on what is administered, the purpose of administration (e.g., prevention or treatment), the patient's state, the mode of administration, and the like. In therapeutic applications, the amount of the composition that can be administered to a patient already suffering from a disease is sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the disease condition being treated and on the judgment of the attending physician depending on factors such as the severity of the disease, the patient's age, weight, and general condition, and the like.

[0218] The composition administered to the patient may be in the form of the pharmaceutical composition described above. These compositions may be sterilized by conventional sterilization techniques, or may be aseptically filtered. The aqueous solution may be packaged for use as is, or lyophilized, and the lyophilized preparation may be combined with a sterile aqueous carrier before administration. The pH value of the compound preparation will generally be between 3 and 11, more preferably 5 to 9 and most preferably 7 to 8. It should be understood that the use of some of the aforementioned excipients, carriers or stabilizers will form a pharmaceutical salt.

[0219] The therapeutic dose of the compounds of the present invention can vary according to, for example, the specific use for treatment, the mode of administration of the compound, the health status and illness of the patient, and the judgment of the prescribing physician. The ratio or concentration of the compounds of the present invention in the pharmaceutical composition can vary depending on many factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention can be provided in the form of an aqueous physiological buffer solution containing about 0.1 w / v% to about 10 w / v% of the compound for parenteral administration. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg body weight per day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg body weight per day. The dosage may depend on variables such as the type and degree of development of the disease or condition, the overall health status of a particular patient, the relative biological efficacy of the selected compound, the formulation of the excipient, and its route of administration. The effective dose can be extrapolated from a dose-response curve derived from an in vitro or animal model test system.

[0220] The compositions of the invention may also include one or more additional agents, such as chemotherapeutic agents, steroids, anti-inflammatory compounds, or immunosuppressive agents, examples of which are listed herein.

[0221] Labeled compounds and assays

[0222] Another aspect of the invention relates to compounds of the invention (e.g., salts and crystalline forms) that are labeled (radioactively labeled, fluorescently labeled, etc.) and that can be used not only for imaging techniques but also for in vitro and in vivo assays for localizing and quantifying PI3K in tissue samples (including humans), and for identifying PI3K ligands by inhibiting the binding of labeled compounds. Substitution of one or more atoms of the compounds of the invention can also be used to generate differential ADME (absorption, distribution, metabolism, and excretion). Thus, the invention includes PI3K assays containing such labeled or substituted compounds.

[0223] The present invention also includes isotopically labeled compounds of the present invention. An "isotopically labeled" or "radiolabeled" compound is a compound of the present invention in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated into the compounds of the present invention include, but are not limited to 2 H (deuterium, also written as D), 3 H (tritium, also written as T), 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 18 F. 35 S. 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I. 124 I. 125 I and 131 I. For example, one or more hydrogen atoms in the compounds of the invention may be replaced by deuterium atoms (e.g., C 1-6 One or more hydrogen atoms of the alkyl group may be optionally replaced by a deuterium atom, such as -CD 3 Substitute -CH 3 ). In some embodiments, the alkyl groups of the disclosed formulae and / or forms may be perdeuterated.

[0224] One or more constituent atoms of the compounds provided herein may be replaced or substituted with an isotope of an atom in natural or non-natural abundance. In some embodiments, the compounds include at least one deuterium atom. For example, one or more hydrogen atoms in the compounds provided herein may be replaced or substituted with deuterium (e.g., C 1-6 One or more hydrogen atoms of an alkyl group may be replaced by a deuterium atom, such as -CD 3 Substitute -CH 3 ). In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1, 1-2, 1-3, 1-4, 1-5, or 1-6 deuterium atoms. In some embodiments, all hydrogen atoms in the compound can be replaced or substituted with deuterium atoms.

[0225] In some embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms attached to the carbon atoms of any of the formula A substituents are each optionally replaced with a deuterium atom.

[0226] Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry, Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange, Jens Atzrodt, Volker Derdau, Thorsten Fey; and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling, James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds can be used in a variety of studies, such as NMR spectroscopy, metabolic experiments and / or assays.

[0227] Substitution with heavier isotopes (such as deuterium) may provide certain therapeutic advantages due to greater metabolic stability, for example, increasing half-life in vivo or reducing dosage requirements, and therefore may be preferred in some cases. (See, for example, A. Kerekes et al., J. Med. Chem. 2011, 54, 201-210; R. Xu et al., J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolic sites may provide one or more therapeutic advantages.

[0228] The radionuclide incorporated into the radiolabeled compounds of the invention will depend on the specific application of the radiolabeled compound. For example, for in vitro PI3K labeling and competition assays, incorporation of 3 H. 14 C. 82 Br, 125 I. 131 I or 35 For radiographic applications, 11 C. 18 F. 125 I. 123 I. 124 I. 131 I. 75 Br, 76 Br or 77 Br may be useful.

[0229] It is understood that a "radiolabel" or "labeled compound" is a compound that incorporates at least one radionuclide. In some embodiments, the radionuclide is selected from 3 H. 14 C. 125 I. 35 S and 82 The group composed of Br.

[0230] The present invention may also include synthetic methods for incorporating radioisotopes into compounds of the present invention.Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and those skilled in the art will readily recognize methods suitable for use with compounds of the present invention.

[0231] The labeled compounds of the present invention can be used in screening assays to identify / evaluate compounds. For example, the ability to bind to PI3K can be assessed by tracking the label to monitor the change in concentration of the newly synthesized or identified compound (i.e., the test compound) when it contacts PI3K. For example, the ability of the test compound (after labeling) to reduce the binding of another compound (i.e., the standard compound) known to bind to PI3K can be assessed. Therefore, the ability of the test compound to compete with the standard compound for binding to PI3K is directly related to its binding affinity. In contrast, in some other screening assays, the standard compound is labeled and the test compound is not labeled. Therefore, the concentration of the labeled standard compound is monitored to assess the competition between the standard compound and the test compound, and thus determine the relative binding affinity of the test compound.

[0232] Pill Box

[0233] The present invention also includes a pharmaceutical kit that can be used, for example, to treat or prevent PI3K-related diseases or disorders (such as cancer), the kit comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a salt of the present invention and a crystalline form. As will be apparent to those skilled in the art, the kit may also include one or more of a variety of traditional pharmaceutical kit components, for example, containers containing one or more pharmaceutically acceptable carriers, other containers, etc., when necessary. The kit may also include instructions in the form of inserts or labels, indicating the amount of the components to be administered, instructions for administration, and / or instructions for mixing the components.

[0234] The present invention will be described in more detail by specific examples. The following examples are provided for illustrative purposes and are not intended to limit the present invention in any way. One skilled in the art will readily recognize multiple non-critical parameters that can be changed or modified to produce substantially the same results. According to at least one assay described herein, it has been found that the salts and crystalline forms of the embodiments are PI3Kγ inhibitors.

[0235] Example

[0236] Preparative LC-MS purification of some of the compounds prepared was performed on a Waters mass directed fractionation system. The basic equipment setup, protocols, and control software for operating these systems have been described in detail in the literature (see, for example, “Two-Pump At Column Dilution Configuration for Preparative LC-MS”, K. Blom, J. Combi. Chem., 4, 295 (2002); “Optimizing Preparative LC-MS Configurations and Methods for Parallel Synthesis Purification”, K. Blom, R. Sparks, J. Doughty, G. Everlof, T. Haque, A. Combs, J. Combi. Chem., 5, 670 (2003); and “Preparative LC-MS Purification: Improved Compound Specific Method Optimization”, K. Blom, B. Glass, R. Sparks, A. Combs, J. Combi. Chem., 6, 874-883 (2004)). The separated compounds were usually subjected to analytical liquid chromatography mass spectrometry (LCMS) for purity analysis under the following conditions: Instrument: Agilent 1100 series, LC / MSD; Column: Waters Sunfire TM C 18 5 μm, 2.1 x 50 mm; buffer: mobile phase A: 0.025% TFA in water and mobile phase B: acetonitrile; gradient 2% to 80% B in 3 minutes, flow rate 2.0 mL / min.

[0237] As also indicated in the Examples, some of the prepared compounds were separated on a preparative scale by reverse phase high performance liquid chromatography (RP-HPLC) or flash chromatography (silica gel) with MS detection. Typical preparative reverse phase high performance liquid chromatography (RP-HPLC) column conditions were as follows:

[0238] pH = 2 Purification: Waters Sunfire TM C 18 5μm, 30x 100mm or Waters XBridge TM C 185 μm, 30 x 100 mm column, eluted with mobile phase A: 0.1% TFA (trifluoroacetic acid) in water and mobile phase B: acetonitrile; the flow rate was 60 mL / min, and the separation gradient of each compound was optimized using the compound-specific method optimization scheme described in the literature (see, for example, "Preparative LCMS Purification: Improved Compound Specific Method Optimization", K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)).

[0239] pH = 10 Purification: Waters XBridge TM C 18 5 μm, 30 x 100 mm column, with mobile phase A: 0.1% NH 4 OH aqueous solution and mobile phase B: acetonitrile were used for elution; the flow rate was 60 mL / min, and the separation gradient of each compound was optimized using the compound-specific method optimization scheme described in the literature (see, for example, "Preparative LCMS Purification: Improved Compound Specific Method Optimization", K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)).

[0240] In the following examples, X-ray powder diffraction analysis was performed on a Bruker D8 Advance ECO X-ray powder diffractometer (XRPD) instrument with the following parameters: the radiation source was Cu, The LYNXEYETM detector and X-ray power were 40KV, 25mA. The sample powder was dispersed on a zero-background sample holder. The general measurement conditions were: starting angle – 3°; ending angle – 30°; sampling – 0.015°; and scanning speed – 2° / min.

[0241] Differential Scanning Calorimetry (DSC) was performed on a TA Instruments Differential Scanning Calorimetry Discovery DSC2500 with autosampler. General experimental conditions were: 20-300°C, 10°C / min, nitrogen flow rate 50 mL / min, using aluminum sample pans.

[0242] Thermogravimetric analysis (TGA) was performed on a TA Instruments Thermogravimetric Analyzer TGA5500 with autosampler under the following conditions: 25°C to 600°C at 10°C / min; nitrogen at 25 mL / min balance purge flow; and platinum sample pan.

[0243] Example 1. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide

[0244]

[0245] Step 1. 1-(3-Bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-ol

[0246]

[0247] A solution of 3-bromo-4-methylbenzaldehyde (6.51 g, 32.7 mmol) [Aldrich, 565334] in tetrahydrofuran (65.4 mL) was cooled to 0 ° C and treated with trimethyl (trifluoromethyl) silane (6.28 mL, 42.5 mmol). The yellow mixture was treated with 1.0 M tetrabutylammonium fluoride (0.654 mL, 0.654 mmol) in tetrahydrofuran at 0 ° C and stirred for several minutes at 0 ° C. The ice bath was removed and the resulting reaction mixture was stirred for 1.5 hours. The reaction mixture was cooled back to 0 ° C and treated with water (6.48 mL, 360 mmol) and 1.0 M tetrabutylammonium fluoride (6.54 mL, 6.54 mmol) in tetrahydrofuran. The ice bath was removed and the reaction mixture was stirred for 30 minutes at ambient temperature. The yellow reaction mixture was diluted with brine (150 mL) and extracted with ethyl acetate (200 mL). The organic layer was washed with saturated ammonium chloride (100 mL), dried over sodium sulfate, filtered and concentrated to give a brown oil. Purification by flash column chromatography using methyl tert-butyl ether (MTBE) in hexanes (0% to 50%) gave the desired product (8.42 g, 95.7%) as a yellow oil. LCMS C 9 H 7 F 3 (M-OH) + :m / z=251.0,253.0; Exp. value: 250.9,252.8.

[0248] Step 2. 1-(3-Bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-one

[0249]

[0250] A mixture of 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-ol (8.41 g, 31.3 mmol) in dichloromethane (125 mL) was treated with Dess-Martin periodinane (19.9 g, 46.9 mmol) at 0°C and stirred at room temperature for 2.5 hours. The reaction mixture was concentrated (by rotary evaporation with the water bath set at 30°C) to an oily solid and diluted with ether (200 mL), which resulted in more solid precipitation. The mixture was filtered and rinsed with additional ether (200 mL). The filtrate was washed with saturated sodium bicarbonate solution (3 x 200 mL) and brine, dried over sodium sulfate, filtered and concentrated to give an oily solid. The oily solid was partitioned between ether (150 mL) and water (100 mL). The organic layer was separated and washed with saturated sodium bicarbonate solution (2 x 75 mL) and brine, dried over sodium sulfate, filtered and concentrated to give the desired product (7.93 g, 95.0%) as an oil, which was used without further purification. LCMS C 9 H 7 F 3 O(M+H) + :m / z=267.0,269.0; Exp. value: 267.1,268.9.

[0251] Step 3. 2-(3-Bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionitrile

[0252]

[0253] A solution of 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-one (7.92 g, 29.7 mmol) in dichloromethane (29.7 mL) was treated with trimethylsilyl cyanide (8.70 mL, 65.2 mmol), potassium cyanide (0.290 g, 4.45 mmol) and 18-crown-6 (0.290 g, 1.10 mmol) and stirred for 1 hour. After the addition of 18-crown-6, the reaction was cooled with an ice bath due to the exotherm. The reaction mixture was concentrated (by rotary evaporation with the water bath set at 28 °C) to give a rust-colored solid. The solid was dissolved in THF (29.6 mL), cooled to 0 °C, treated with 1.8 M HCl (10.9 mL, 19.6 mmol), and stirred at room temperature (rt) for 1.5 hours. The reaction mixture was diluted with water (75 mL) and extracted with diethyl ether (3 x 75 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated. Reconcentration from hexanes gave the desired product as an orange solid (8.70 g, 99.8%) which was used without further purification. LCMS C9 H 7 F 3 O(M-CN) + :m / z=267.0,269.0; Exp. value: 266.9,269.0.

[0254] Step 4. 2-(3-Bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (second eluting mirror image isomer)

[0255]

[0256] A solution of 2-(3-bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionitrile (8.70 g, 29.6 mmol) in 1,4-dioxane (59.2 mL) was treated at 0°C with concentrated HCl (9.00 mL, 108 mmol) pre-cooled in an ice bath. The reaction mixture was bubbled with HCl gas for 45 minutes while stirring at 0°C. The cooling bath was removed and the reaction mixture was stirred at room temperature for 61 hours. The reaction mixture was bubbled with nitrogen for 10 minutes to remove some HCl, cooled to 0°C, and diluted with brine (200 mL), water (50 mL), and ethyl acetate (200 mL). The organic layer was separated and the aqueous layer was diluted with water (100 mL) to dissolve the remaining solids. The aqueous layer was extracted with ethyl acetate (100 mL). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated to give a brown oil. By flash column chromatography, MTBE (0% to 60%) in hexane is used to purify to obtain a racemic product in the shape of a yellow oily solid. The racemic mixture is separated via preparative chiral HPLC (Phenomenex Lux Amylose-1 [21.2x250mm, 5 μm], eluted with 95% ethanol in hexane at a flow rate of 18mL / min, about 100mg is loaded in 2mL ethanol) to obtain the required second eluting mirror image isomer (4.50g, 48.8%) in a viscous yellow oil. The residence time of the first mirror image isomer eluted is 4.0 minutes. The residence time of the second mirror image isomer eluted is 5.3 minutes.

[0257] Second eluting mirror image isomer : 1 H NMR (400 MHz, DMSO-d 6 )δ7.85(d,J=1.9Hz,1H),7.75(s,1H),7.67(s,1H),7.63–7.53(m,2H),7.41(d,J=8.1Hz,1H),2.35(s,3H). LCMSC 10 H 10 F 3 NO 2(M+H) + :m / z=312.0,314.0; Exp. value: 312.0,314.0.

[0258] Step 5. 3,3,3-Trifluoro-2-hydroxy-2-(4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanamide

[0259]

[0260] A solution of 2-(3-bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (3.57 g, 11.5 mmol) (Example 1, Step 4, second eluting mirror image isomer) in 1,4-dioxane (57.2 mL) was treated with bis(pinacolato)diboron (3.49 g, 13.7 mmol) and potassium acetate (3.71 g, 37.8 mmol) and degassed with nitrogen for 5 minutes. The reaction mixture was treated with bis(triphenylphosphine)palladium(II) chloride (0.482 g, 0.687 mmol), degassed for 5 minutes, and stirred at 100° C. for 2.5 hours. The reaction mixture was diluted with ethyl acetate (50 mL) and heated to 40° C. for 1 h. The 1-(2-nitro-1-yl)-2-nitropropene was stirred for 2 hours at 4 0 ℃ for 1 h.Then the mixture was stirred for 2 hours at 4 0 ℃ for 3 hours ...Filter and rinse with additional ethyl acetate (100 mL).The filtrate was washed with brine, dried over sodium sulfate, filtered, and concentrated into a brown foam.Purified by flash column chromatography using MTBE (0% to 100%) in hexanes to give the desired product (3.35 g, 81.5%) as a thick yellow foam. 1 HNMR (400 MHz, DMSO-d 6 )δ7.96(d,J=2.2Hz,1H),7.63(dd,J=7.9,2.1Hz,1H),7.58(s,1H),7.54(s ,1H),7.51–7.40(m,1H),7.21(d,J=8.2Hz,1H),2.46(s,3H),1.30(s,12H). LCMSC 16 H 22 BF 3 NO 4 (M+H) + :m / z=360.2; Exp. value: 360.1.

[0261] Step 6. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide

[0262] A solution of 3-bromo-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-8-amine (7.50 g, 26.7 mmol) and 3,3,3-trifluoro-2-hydroxy-2-(4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanamide (10.5 g, 29.4 mmol) (Example 1, Step 5) in 1,4-dioxane (133 mL) was treated with 1.0 M aqueous potassium carbonate solution (53.4 mL, 53.4 mmol), degassed with nitrogen for 5 minutes, treated with dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloromethane adduct (3.27 g, 4.00 mmol), degassed with nitrogen for another 5 minutes, and stirred at 100°C for 19 hours. The reaction mixture was treated with ethyl acetate (200 mL) and brine (50 mL), filtered through Celite and rinsed with additional ethyl acetate. The aqueous layer was separated from the filtrate and extracted with ethyl acetate (200 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated into brown foam. Purification was performed using MeOH (0% to 10%) in dichloromethane by flash column chromatography to obtain the desired product in red / brown foam, which was not completely pure. Purification of the substance using MeOH (0% to 15%) in dichloromethane by flash column chromatography gave the desired product in orange / brown foam, which was still not completely pure. Purification of the substance using ethyl acetate (containing 5% MeOH) (0% to 100%) in hexane by flash column chromatography gave the desired product in white foam, which still contained impurities. This material was repurified by flash column chromatography using acetonitrile (5% MeOH) in dichloromethane (0% to 100%) to give the desired product (4.67 g, 40.4%) as a white foam. 1 H NMR (600 MHz, DMSO-d 6 )δ7.79(s,1H),7.76-7.71(m,2H),7.71-7.64(m,4H),7.61(d,J=3.5Hz,2H),7.51(d,J=8.2Hz,1H),2.23(s,3H). LCMSC 17 H 14 F 6 N 5 O 2 (M+H) + :m / z=434.1; Exp. value: 434.1.

[0263] Example 2: Preparation and Characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide Crystalline Form IA (Free Base)

[0264] A vial was charged with 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (0.050 g, 0.115 mmol) and stirred at 80°C while a 1:2 premixed solution of isopropyl acetate (0.676 mL) / heptane (1.34 mL) was added dropwise. After adding 2 mL, the solid was not completely dissolved and some remained at the bottom of the vial. After almost all of the solid was dissolved, new solid formed on the walls of the vial. More solid formed after stirring at 80°C for 2 hours. After cooling to ambient temperature, the solid was filtered and washed with heptane. The solid was collected and dried under reduced pressure for 30 min to give 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (Form IA) (33.2 mg, 66.4%) as a white solid.

[0265] Form IA was confirmed to be a crystalline solid based on XRPD analysis. Figure 1 The XRPD pattern of Form IA is shown in and the peak data is provided in Table 1 below.

[0266] Table 1. XRPD peak data for Form IA.

[0267]

[0268]

[0269] DSC analysis of Form IA showed one endothermic peak with an onset temperature of 191.9 °C and a maximum at 193.2 °C. Figure 2 The DSC thermograms are provided in .

[0270] TGA analysis of Form IA showed significant weight loss above 200°C due to sample decomposition. Figure 3 TGA thermograms are provided in .

[0271] Form IA was confirmed to be an anhydrous, non-solvated crystalline form.

[0272] Example 3: Preparation and Characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide Crystalline Form IIA (Free Base)

[0273] Approximately 100 mg of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide free base was dissolved in 1 mL of isopropyl acetate in a 4 mL clear glass vial. 2 mL of heptane was added to the solution with stirring at ambient temperature. The mixture was heated at 80° C. with stirring for 2 hours. The mixture was allowed to cool to ambient temperature and stirred for 1 hour. The solid was collected by filtration and air dried to give 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (Form IIA).

[0274] Form IIA was confirmed to be a crystalline solid based on XRPD analysis. Figure 4 The XRPD pattern of Form IIA is shown in and the peak data is provided in Table 2 below.

[0275] Table 2. XRPD peak data for Form IIA.

[0276]

[0277]

[0278] DSC analysis of Form IIA showed one endothermic peak with an onset temperature of 177.2 °C and a maximum at 179.7 °C. Figure 5 The DSC thermograms are provided in .

[0279] TGA analysis of Form IIA showed significant weight loss above 200°C due to sample decomposition. Figure 6 TGA thermograms are provided in .

[0280] Form IIA was confirmed to be an anhydrous, non-solvated crystalline form.

[0281] Example 4: Preparation and Characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide Crystalline Form IIIA (Free Base)

[0282] Approximately 72 mg of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide free base was dissolved in 1 mL of MeOH in a 4 mL clear glass vial. The solution was evaporated to dryness at ambient temperature. The resulting solid (MeOH solvate) was dried under vacuum at 60 °C overnight to give 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (Form IIIA).

[0283] Form IIIA was confirmed to be a crystalline solid based on XRPD analysis. Figure 7 The XRPD pattern of Form IIIA is shown in and the peak data is provided in Table 3 below.

[0284] Table 3. XRPD peak data for Form IIIA.

[0285]

[0286]

[0287] DSC analysis of Form IIIA showed one endothermic peak with an onset temperature of 134.3 °C and a maximum at 143.0 °C. Figure 8 The DSC thermograms are provided in .

[0288] TGA analysis of Form IIIA showed significant weight loss above 200°C due to sample decomposition. Fig. 9 TGA thermograms are provided in .

[0289] Form IIIA was confirmed to be an anhydrous, non-solvated crystalline form.

[0290] Example 5 to Example 6. 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide (mirror isomer 1 and mirror isomer 2)

[0291]

[0292] Step 1. 6,8-Dibromo-3-iodoimidazo[1,2-a]pyrazine

[0293]

[0294] To a solution of 6,8-dibromoimidazo[1,2-a]pyrazine (0.50 g, 1.8 mmol) [Combi-Blocks, OR-7964] in DMF (12 mL) was added N-iodosuccinimide (0.45 g, 2.0 mmol). The reaction mixture was then heated at 60 °C for 15.5 hours. The reaction mixture was concentrated in vacuo. The resulting solid was dissolved in dichloromethane (DCM). The mixture was washed with water and saturated Na 2 S 2 O 3 (aqueous solution) and then the organic layer was washed with Na 2 SO 4 Drying, filtration and concentration gave the title compound as a light yellow solid (0.64 g, 88%). LCMS C 6 H 3 Br 2 IN 3 (M+H) + : Calculated value m / z = 401.8, 403.8, 405.8; Exp. value 401.8, 403.7, 405.6.

[0295] Step 2. 6-Bromo-3-iodo-N-(4-methoxybenzyl)imidazo[1,2-a]pyrazin-8-amine

[0296]

[0297] A solution of 6,8-dibromo-3-iodoimidazo[1,2-a]pyrazine (1.67 g, 3.57 mmol), N,N-diisopropylethylamine (1.24 mL, 7.13 mmol) and (4-methoxyphenyl)methylamine (0.512 mL, 3.92 mmol) in iPrOH (11.9 mL) was heated in a microwave at 110 °C for 1 hour. After cooling to room temperature, the solidified reaction mixture was diluted with isopropanol (75 mL) and water (19 mL) and stirred for 10 minutes. The solid was collected by filtration to give the desired product (1.41 g, 86.1%), which was used without further purification. LCMS C 14 H 13 BrN 4 O(M+H) + : Calculated value m / z = 458.9, 460.9; Found value 459.0, 461.0.

[0298] Step 3. 6-Bromo-3-iodoimidazo[1,2-a]pyrazine-8-amine trifluoroacetate

[0299]

[0300] A solution of 6-bromo-3-iodo-N-(4-methoxybenzyl)imidazo[1,2-a]pyrazine-8-amine (2.72 g, 5.92 mmol) in trifluoroacetic acid (TFA, 14.8 mL) was stirred at 55 ° C for 5.5 hours. The reaction mixture was concentrated and reconcentrated after dilution with acetonitrile (2x). The solid was diluted with ethyl acetate (12 mL) and stirred at room temperature for 1 hour. The slurry was diluted dropwise with hexane (12 mL) and stirred at room temperature for 75 minutes. The solid was collected by filtration to give the desired product (2.03 g, 75.7%), which was used without further purification. LCMS C 6 H 5 BrN 4 (M+H) + : Calculated value m / z = 338.9, 340.9; Exp. value 338.8, 340.8.

[0301] Step 4. 2-(3-Bromo-4-methylphenyl)-1,1,1-trifluoropropan-2-ol

[0302]

[0303] A solution of 1-(3-bromo-4-methylphenyl) ethyl-1-ketone (1.20 g, 5.63 mmol) [Aldrich, 579734] in tetrahydrofuran (22.5 mL) was treated with trimethyl (trifluoromethyl) silane (1.00 mL, 6.76 mmol) [Aldrich, 488712] at 0 ° C and stirred for 5 minutes at 0 ° C. The reaction mixture was treated with 1.0 M tetra-n-butylammonium fluoride (0.282 mL, 0.282 mmol) in tetrahydrofuran at 0 ° C, and stirred at room temperature for 1 hour. The reaction mixture was cooled to 0 ° C, treated with additional 1.0 M tetra-n-butylammonium fluoride (6.76 mL, 6.76 mmol) in tetrahydrofuran, and stirred at room temperature for 30 minutes. The reaction mixture was diluted with ethyl acetate (100 mL) and washed with brine (2 x 75 mL). The organic layer was separated, dried over sodium sulfate, filtered and concentrated to give a crude residue. Purification by flash column chromatography using ethyl acetate in hexanes (0%-30%) gave the desired product (1.54 g, 96.7%) as a yellow oil. LCMS C 10 H 9 F 3 (M-OH) + :m / z=265.0,267.0; Exp. value: 264.9,267.0.

[0304] Step 5. 1,1,1-Trifluoro-2-(4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ol

[0305]

[0306] A mixture of 2-(3-bromo-4-methylphenyl)-1,1,1-trifluoropropan-2-ol (0.252 g, 0.890 mmol), bis(pinacolyl)diboron (0.294 g, 1.16 mmol) and potassium acetate (0.288 g, 2.94 mmol) in tetrahydrofuran (4.95 mL) was degassed with nitrogen for 5 minutes. The reaction mixture was treated with triphenylphosphine palladium chloride (0.025 g, 0.036 mmol), degassed with nitrogen for 5 minutes, and heated in a microwave at 135 ° C for 20 minutes. The reaction mixture was diluted with ethyl acetate and filtered by means of a 0.5 μm filter cartridge (rinsed with ethyl acetate). The filtrate was washed with water and brine, dried over sodium sulfate, filtered and concentrated to give a crude residue. Purification by flash column chromatography using ether in hexane (0%-50%) gave the desired product (272 mg, 92.5%) as a colorless oil. LCMS C 16 H 23 BF 3 O 3 (M+H) + :m / z=331.2; Exp. value: 331.2.

[0307] Step 6. 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-1,1,1-trifluoropropan-2-ol

[0308]

[0309] A mixture of 6-bromo-3-iodoimidazo[1,2-a]pyrazine-8-amine trifluoroacetate (Step 3, 0.855 g, 1.89 mmol), 1,1,1-trifluoro-2-(4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ol (Step 5; 0.623 g, 1.89 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.131 g, 0.113 mmol) in ethanol (12.6 ml) was treated with 2.0 M aqueous sodium carbonate (1.89 ml, 3.77 mmol), degassed with nitrogen for 5 minutes, and heated in a microwave reactor at 130° C. for 2 hours. The reaction mixture was partially concentrated to remove ethanol and diluted with ethyl acetate and water. The solids were removed by filtration, and the aqueous layer of the filtrate was separated and extracted with ethyl acetate (2×). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and concentrated to give a crude residue. Purification by flash column chromatography using methanol in dichloromethane (0%-2%) gave the desired product (610 mg, 77.8%) as a white foam. LCMS C 16 H 15 F 3 N 4 O(M+H) + :m / z=415.0,417.0; Exp. value: 415.0,417.0.

[0310] Step 7. 8-amino-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid methyl ester

[0311]

[0312] A solution of 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-1,1,1-trifluoropropan-2-ol (Step 6; 0.250 g, 0.602 mmol) in methanol (16.1 ml) was treated with triethylamine (0.336 ml, 2.41 mmol) and degassed with nitrogen for 5 min. 2 CH 2 Cl 2The reaction mixture was treated with 4-(4-(2-nitro-1-yl)-2-nitropropene (0.049 g, 0.060 mmol), degassed with nitrogen for 5 minutes, saturated with CO by bubbling it through the surface of the reactant for 3 minutes, and heated at 60 ° C overnight. The reaction mixture was concentrated and the resulting red oil was diluted with ethyl acetate, water and saturated sodium bicarbonate. The aqueous layer was separated and re-extracted with ethyl acetate (2x). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and concentrated to give a brown oil. Purification by flash column chromatography using methanol (0%-4%) in dichloromethane gave the desired product (158 mg, 66.5%) as an amber oily solid. LCMS C 18 H 18 F 3 N 4 O 3 (M+H) + :m / z=395.1; Exp. value: 395.1.

[0313] Step 8. 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide

[0314] A solution of methyl 8-amino-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylate (Step 7, 0.080 g, 0.203 mmol) in THF (3.38 mL) was treated with 1-amino-2-methylpropan-2-ol (0.181 g, 2.03 mmol) followed by trimethylaluminum (0.507 mL, 1.01 mmol) (2 M in toluene) and stirred at 80° C. overnight. The reaction mixture was treated with additional trimethylaluminum (0.70 ml, 1.40 mmol) (2 M in toluene) and stirred at 80° C. overnight. The reaction mixture was cooled to room temperature, diluted with methanol, and purified by chromatography on HPLC. The mixture was stirred for 2 hours at 4 ℃ for 10 minutes.Then, add 4-nitropropene (2-nitropropene) (4-nitropropene) (5-nitropropene) (6-nitropropene) (7-nitropropene) (8-nitropropene) (10-nitropropene) (20-nitropropene) (10-nitropropene) (20-nitropropene) (5-nitropropene) (6-nitropropene) (10-nitropropene) (20-nitropropene) (10-nitropropene) (20-nitropropene) (5-nitropropene) (6-nitropropene) (8-nitropropene) (10-nitropropene) (20-nitropropene) (10-nitropropene) (20-nitropropene) (5-nitropropene) (6-nitropropene) (8-nitropropene) (10-nitropropene) (20-nitropropene) (20-nitropropene) (10-nitropropene) (20-nitropropene) (20-nitropropene) (

[0315] Embodiment 5 (mirror image isomer 1): 1 H NMR (400 MHz, DMSO-d 6 )δ8.08(t,J=6.1Hz,1H),7.70(d,J=2.8Hz,2H),7.65(d,J=8.1Hz,1H),7.58(s,1H),7.49(d,J=8.2Hz,1H ),7.36(s,2H),6.65(s,1H),4.65(s,1H),3.22(d,J=6.1Hz,2H),2.15(s,3H),1.70(s,3H),1.09(s,6H). LCMS C 21 H 25 F 3 N 5 O 3 (M+H) + :m / z=452.2; Exp. value: 452.1.

[0316] Embodiment 6 (mirror image isomer 2): 1 H NMR (600 MHz, DMSO-d 6 )δ8.08(t,J=6.0Hz,1H),7.70(d,J=3.0Hz,2H),7.65(d,J=8.2Hz,1H),7.58(s,1H),7.49(d,J=8.2Hz,1H ),7.36(s,2H),6.65(s,1H),4.65(s,1H),3.22(d,J=6.1Hz,2H),2.15(s,3H),1.70(s,3H),1.09(s,6H). LCMS C 21 H 25 F 3 N 5 O 3 (M+H) + :m / z=452.2; Exp. value: 452.2.

[0317] Example 7: Preparation and Characterization of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide Crystalline Form IB (Free Base)

[0318] A round bottom flask was charged with 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide (enantiomer 2 from step 8 of Example 6; 184 g, 408 mmol) and isopropyl acetate (950 mL). The mixture was stirred at 80° C. for 1 hour, cooled to room temperature (RT), and stirred at room temperature overnight. The solid was collected to give 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide (Form IB, 152 g, 82.8%).

[0319] Form IB was confirmed to be a crystalline solid based on XRPD analysis. Fig.10 The XRPD pattern of Form IB is shown in and the peak data is provided in Table 4 below.

[0320] Table 4. XRPD peak data for Form IB.

[0321]

[0322]

[0323] DSC analysis of Form IB showed one endothermic peak with an onset temperature of 172.2 °C and a maximum at 174.2 °C. Fig.11 A DSC thermogram is provided in . It was confirmed that Form IB is an anhydrous non-solvated crystalline form.

[0324] Example 8: Preparation and Characterization of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide Crystalline Form IIB (Free Base)

[0325] A vial was charged with 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide (enantiomer 2 from Example 6, Step 8; 252 mg, 0.559 mmol) and isopropyl acetate (1.25 mL) and the solid was slowly dissolved. The mixture was treated with heptane (0.35 mL) until the solid did not change. The mixture was heated at 80° C. for 30 minutes and stirred at room temperature overnight. The solid was collected to give 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide (Form IIB, 116 mg, 46.0%).

[0326] Form IIB was confirmed to be a crystalline solid based on XRPD analysis. Fig.12 The XRPD pattern of Form IIB is shown in and the peak data are provided in Table 5 below.

[0327] Table 5. XRPD peak data for Form IIB.

[0328]

[0329]

[0330] DSC analysis of Form IIB showed one endothermic peak with an onset temperature of 161.7 °C and a maximum at 165.4 °C. Figure 8 The DSC thermogram is provided in . It was confirmed that Form IIB is an anhydrous non-solvated crystalline form.

[0331] Example 9. 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide

[0332]

[0333] Step 1. 1-(4-(methyl-d 3 )phenyl)ethan-1-one

[0334]

[0335] A solution of (4-acetylphenyl)boronic acid (1.00 g, 6.10 mmol) [Aldrich, 470821], bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II) (0.108 g, 0.152 mmol) and cesium fluoride (3.24 g, 21.4 mmol) in DMF (10.2 mL) and water (2.03 mL) was degassed with nitrogen for 10 min and then heated with iodomethane-d 3 To the 4-thiazolinyl esters (1.44mL, 23.2mmol) was added 4-thiazolinyl esters (1.44mL, 23.2mmol) and stirred at 45°C overnight. The reaction mixture was cooled to room temperature and diluted with water and ethyl acetate. The aqueous layer was separated and extracted with ethyl acetate (2x). The combined organic extracts were washed with water and brine, dried over magnesium sulfate, filtered and concentrated (60-70 Torr, 25°C bath) to obtain the desired product (546mg, 65.3%) as a yellow oil, which was not further purified and used. 1 H NMR (400 MHz, CDCl 3 )δ7.86(d,J=8.0Hz,2H),7.26(d,J=8.0Hz,2H),2.58(s,3H). LCMS C 9 H 8 D 3 O(M+H) + :m / z=138.1; Exp. value: 138.1.

[0336] Step 2. 1-(3-Bromo-4-(methyl-d 3 )phenyl)ethan-1-one

[0337]

[0338] 1-(4-(methyl-d 3) phenyl) second -1- ketone (6.35g, 46.3mmol) is dropwise treated with aluminum chloride (13.6g, 102mmol) suspension in dichloromethane (24mL). The residual material in the syringe is rinsed with dichloromethane (7.0mL) and added dropwise to the reaction mixture. After the initial heat release, the reaction mixture is allowed to cool to room temperature and kept for 3 minutes, stirred at 35 ℃ for 5 minutes, and treated dropwise with bromine (2.38mL, 46.3mmol) within 5 minutes. The reaction mixture is stirred for 25 minutes and then slowly added to a mixture of dichloromethane (50mL), 1N HCl (100mL) and ice. The residual reaction mixture is rinsed into a dichloromethane / HCl / ice mixture with additional dichloromethane. The mixture is warmed to room temperature (rt) and the layers are separated. The aqueous layer is extracted with dichloromethane (2x 75mL). The combined organic layer is washed with saturated sodium bicarbonate and brine. Sodium bicarbonate and brine washings contain product and these washings are combined, acidified with 1M HCl, and extracted with dichloromethane (2x 50mL). Combine all organic layers, dry over magnesium sulfate, filter and concentrate to obtain a yellow oil. Purify using ethyl acetate (0%-15%) in hexane to obtain the desired product (9.08g, 90.8%) as a light yellow solid by flash column chromatography. 1 H NMR (400 MHz, CDCl 3 )δ8.11(d,J=1.8Hz,1H),7.79(dd,J=7.9,1.8Hz,1H),7.32(d,J=7.9Hz,1H),2.57(s,3H). LCMS C 9 H 7 D 3 BrO(M+H) + :m / z=216.0,218.0; Exp. value: 216.0,218.0.

[0339] Step 3. 2-(3-Bromo-4-(methyl-d 3 )phenyl)-1,1,1-trifluoropropan-2-ol

[0340]

[0341] 1-(3-Bromo-4-(methyl-d)-1-((((trifluoromethyl)-1-(((trimethyl)-1-(((trifluoro ... 3) phenyl) second -1- ketone (9.08g, 42.0mmol) in tetrahydrofuran (168mL) solution and stirred at 0 ℃ for 5 minutes. At 0 ℃, the reaction mixture was treated with 1.0M tetra-n-butylammonium fluoride (2.10mL, 2.10mmol) in tetrahydrofuran, and stirred at room temperature for 1 hour. The reaction mixture was treated with 1.0M tetra-n-butylammonium fluoride (12.6mL, 12.6mmol) and water (9.8mL) in tetrahydrofuran, and stirred at room temperature for 30 minutes. The reaction mixture was diluted with water (100mL) and extracted with ethyl acetate (3x 50mL). The combined organic layer was washed with brine, dried over magnesium sulfate, filtered and concentrated to obtain a crude residue. Purified using ethyl acetate (0%-20%) in hexane by flash column chromatography to obtain the desired product (13.3g, 111%) in a yellow oily state. 1 H NMR (400 MHz, CDCl 3 )δ7.79–7.72(m,1H),7.48–7.35(m,1H),7.24(s,1H),2.41(br s,1H),1.76(s,3H).

[0342] Step 4. 1,1,1-trifluoro-2-(4-(methyl-d 3 )-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ol

[0343]

[0344] With 2-(3-bromo-4-(methyl-d 3 A suspension of bis(pinacolato)diboron (12.8 g, 50.2 mmol) and potassium acetate (8.63 ml, 138 mmol) in dioxane (24 mL) was treated with 1,2-difluoro-2-(4-(4-phenyl)-1,1,1-trifluoropropan-2-ol (13.3 g, 41.8 mmol). The residual 2-(3-bromo-4-(methyl-d 3 )phenyl)-1,1,1-trifluoropropan-2-ol and added to the reaction mixture and degassed with nitrogen for 10 minutes. The reaction mixture was treated with bis(triphenylphosphine)palladium(II) dichloride (1.16 g, 1.67 mmol), degassed with nitrogen for another 10 minutes, and stirred at 100°C overnight. The reaction mixture was cooled to room temperature, degassed with nitrogen for 5 minutes, treated with additional bis(triphenylphosphine)palladium(II) dichloride (1.16 g, 1.67 mmol), degassed with nitrogen for another 5 minutes, and stirred at 100°C for 4 hours. The reaction mixture was allowed to react with 4% bis(triphenylphosphine)palladium(II) dichloride (1.16 g, 1.67 mmol), degassed with nitrogen for another 5 minutes, and stirred at 100°C for 4 hours. The product was filtered and rinsed with THF and ethyl acetate. The filtrate was washed with 1: 1 water / brine (300 mL). The aqueous layer was extracted with ethyl acetate. The combined organic layer was dried over magnesium sulfate, filtered and concentrated into a brown oil. Purification was performed using MTBE (0%-20%) in hexane by flash column chromatography to obtain the desired product (14.4 g, 84.7%) as a light yellow oil. 1 H NMR (400 MHz, CDCl 3 )δ7.93(d,J=2.3Hz,1H),7.55–7.45(m,1H),7.19(d,J=8.1Hz,1H),2.43(br s,1H),1.77(s,3H),1.34(s,12H). LCMS C 16 H 20 D 3 BF 3 O 3 (M+H) + :m / z=334.2; Exp. value: 334.3.

[0345] Step 5. 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d 3 )phenyl)-1,1,1-trifluoropropan-2-ol (racemic mixture)

[0346]

[0347] 1,1,1-trifluoro-2-(4-(methyl-d 3 )-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ol (14.5 g, 35.6 mmol) in dioxane (178 mL) was degassed with nitrogen for 5 minutes, treated with dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloromethane adduct (5.81 g, 7.11 mmol) and degassed with nitrogen for another 5 minutes. The reaction mixture was treated with 1.0 M aqueous potassium carbonate solution (107 ml, 107 mmol), degassed with nitrogen for 5 minutes, and stirred at 80° C. overnight. The reaction mixture was cooled to room temperature and heated to 40° C. for 1 h. Filter. Rinse with ethyl acetate and water The filtrate was diluted with water (150 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated to a dark oil. Purification by flash column chromatography using methanol in dichloromethane (0%-5%) and repurification by flash column chromatography using ethyl acetate in hexanes (0%-100%) gave the desired product (13.8 g, 92.8%). LCMS C 16 H 12 D 3 F 3 N 4 O(M+H) + :m / z=418.1,420.1; Exp. value: 418.0,420.0.

[0348] Step 6. 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d 3 The second eluting mirror image isomer of 2-(phenyl)-1,1,1-trifluoropropan-2-ol

[0349]

[0350] 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d 3 The retention time of the first peak eluted was 9.6 minutes. The retention time of the second peak eluted was 14.6 minutes.

[0351] Peak 2 : 1 H NMR (400 MHz, DMSO-d 6 )δ7.66–7.59(m,2H),7.59–7.53(m,3H),7.46(d,J=8.1Hz,1H),7.25(s,1H),6.66(s,1H),1.71(s,3H). LCMS C 16 H 12 D 3 F 3 N 4 O(M+H) + :m / z=418.1,420.1; Exp. value: 418.0,420.0.

[0352] Step 7. 8-amino-3-(2-(methyl-d 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid methyl ester (prepared single mirror image isomer)

[0353]

[0354] A solution of 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol (Peak 2 from Step 6, 4.95 g, 48.9 mmol) in methanol (163 mL) and DMF (40.7 mL) was treated with triethylamine (6.81 mL, 48.9 mmol) and degassed with nitrogen for 5 min. Pd(dppf) 2 CH 2 Cl 2 The reaction mixture was treated with 4-(4-(2-nitro-1-yl)-2-nitropropene (0.998 g, 1.22 mmol), degassed with nitrogen for 5 minutes, saturated with CO by bubbling through the surface of the reactant for 3 minutes, and heated at 60 ° C overnight. The reaction mixture was concentrated and the resulting oil was diluted with ethyl acetate and water. The aqueous layer was separated and re-extracted with ethyl acetate (3x). The combined organic layers were washed with water, saturated ammonium chloride solution and brine, dried over magnesium sulfate, filtered and concentrated to a brown oil. Purification by flash column chromatography using methanol (0%-5%) in dichloromethane gave the desired product (4.49 g, 92.4%) as an orange solid. LCMS C 18 H 15 D 3 F 3 N 4 O 3 (M+H) + :m / z=398.1; Exp. value: 398.3.

[0355] Step 8. 8-amino-3-(2-(methyl-d 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid (prepared single mirror image isomer)

[0356]

[0357] 8-amino-3-(2-(methyl-d 3)-5-(1,1,1-trifluoro-2-hydroxypropyl-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid methyl ester (4.49 g, 11.3 mmol) (single mirror image isomer from step 7) in methanol (113 mL) and stirred at room temperature. The reaction mixture was concentrated to remove methanol, diluted with water (50 mL), and extracted with ethyl acetate (50 mL, then 20 mL). The combined ethyl acetate layer was extracted with additional 1.0 M sodium hydroxide (3x20 mL). The combined alkaline aqueous layer was adjusted to pH about 5 with citric acid (7.6 g). The aqueous layer was extracted with dichloromethane (2x 150 mL). The aqueous layer was diluted with brine and extracted with ethyl acetate (150 mL). The combined organic layers were concentrated to give the desired product (4.06 g, 93.8%) as a brown solid, which was used without further purification. 1 H NMR (400 MHz, DMSO-d 6 )δ7.76(s,1H),7.71(s,1H),7.65(dd,J=8.2,2.0Hz,1H),7.59(d,J=2.0Hz,1H),7.49(d,J=8.1Hz,1H),7.30(br s,2H),6.66(s,1H),1.71(s,3H). LCMSC 17 H 13 D 3 F 3 N 4 O 3 (M+H) + :m / z=384.1; Exp. value: 384.2.

[0358] Step 9. 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide

[0359] 8-Amino-3-(2-(methyl-d)-propan-2-ol (1.44 g, 16.2 mmol) [Ark Pharm, AK-37803] and HATU (6.16 g, 16.2 mmol) were treated with 3)-5-(1,1,1-trifluoro-2-hydroxypropyl-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid (4.06 g, 10.6 mmol) (single mirror image isomer from step 8) in DMF (106 mL), stirred for 15 minutes, treated with triethylamine (4.43 mL, 31.8 mmol) and stirred at room temperature for 3.5 hours. The reaction mixture was diluted with water (500 mL) and brine (100 mL) and extracted with ethyl acetate (3 x 150 mL). The combined organics were washed with saturated ammonium chloride (150 mL), 11% sodium carbonate (150 mL) and brine (100 mL), dried over magnesium sulfate, filtered and concentrated to an amber oil. Purification by flash column chromatography using methanol (0%-5%) in dichloromethane gave the desired product (4.28 g, 89.0%) in a foamy state. 1 H NMR (600 MHz, DMSO-d 6 )δ8.14–8.05(m,1H),7.74–7.69(m,2H),7.66(d,J=7.9Hz,1H),7.62–7.54(m,1H),7.50(dd,J=8.2, 2.0Hz, 1H), 7.38 (s, 2H), 6.67 (s, 1H), 4.67 (s, 1H), 3.23 (d, J = 5.6Hz, 2H), 1.71 (s, 3H), 1.10 (s, 6H). LCMS C 21 H 22 D 3 F 3 N 5 O 3 (M+H) + :m / z=455.2; Exp. value: 455.2.

[0360] Example 10: Preparation and Characterization of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide Crystalline Form IC (Free Base)

[0361] The round bottom was loaded with 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide (4.60 g, 10.1 mmol) and isopropyl acetate (25.5 mL) were added and heated at 80°C. The mixture was stirred at 80°C and solids began to form within 5 minutes. The mixture was stirred at 80°C for 1 hour. The heating was interrupted and the mixture was stirred for 1 hour while cooling to room temperature. The mixture was treated dropwise with heptane (25.5 mL) from an addition funnel over 35 minutes and stirred at room temperature for 40 minutes. The solid was collected, washed with 1:1 isopropyl acetate / heptane (10 mL) and dried at 60°C under reduced pressure for 24 hours to give 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide (Form I) (4.16 g, 90.4%).

[0362] Form IC was confirmed to be a crystalline solid based on XRPD analysis. Fig.14 The XRPD pattern of Form IC is shown in and the peak data is provided in Table 6 below.

[0363] Table 6. XRPD peak data for Form IC.

[0364]

[0365]

[0366] DSC analysis of Form IC showed one endothermic peak with an onset temperature of 173.4 °C and a maximum at 179.0 °C. Fig.15 The DSC thermogram is provided in . It was confirmed that Form IC is an anhydrous non-solvated crystalline form.

[0367] Example 11. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromic acid (HBr) salt

[0368] 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide free base (98.81 mg) was dissolved in 2.5 mL of methanol in a 4 mL clear glass vial. 42.4 μL of 6M HBr aqueous solution (1.2 eq) was added to the solution and mixed thoroughly. The solution was evaporated at room temperature to obtain HBr salt crystals.

[0369] Example 12. Characterization of Single Crystal 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide (HBr) salt

[0370] Crystal data :C35 H32 Br2 F12 N10 O5, from methanol, colorless, irregular flakes, about 0.450x0.210x 0.060mm, monoclinic, C2, β=94.953(7), Z = 4, T = -40 ° C, formula weight = 1060.52, density = 1.631 g / cm 3 ,μ(Mo)=1.98mm -1 .

[0371] Data Collection Data collection was performed as follows: using a Bruker SMART APEX-II CCD system, MoKα radiation, standard focusing tube, anode power = 50 kV x 30 mA, crystal to plate distance = 5.0 cm, 512 x 512 pixels / frame, beam center = (259.19, 253.13), total number of frames = 2635, oscillation / frame = 0.50°, exposure / frame = 40.1 sec / frame, SAINT integration, hkl min / max = (-26, 26, -12, 13, -27, 27), data input to shelx = 38968, unique data = 9756, 2θ range = 4.51° to 55.43°, 2θ55.43 integrity = 99.60%, R(int-xl) = 0.0672, SADABS correction applied.

[0372] Parsing and Refinement :The crystal structure was solved using XS (Shelxtl) and refined using the shelxtl package. Refinement was performed by F 2 Full matrix least squares, scattering factors from Int. Tab. Vol C Table 4.2.6.8 and Table 6.1.1.4, number of data = 9756, number of constraints = 1, number of parameters = 584, data / parameter ratio = 16.71, F2 goodness of fit = 1.14, R index [I>4sigma (I)] R1 = 0.0648, wR2 = 0.1560, R index (all data) R1 = 0.1004, wR2 = 0.1719, maximum difference peaks and holes = 1.795 and , refined flack parameter = 0.038 (6). All hydrogen atoms were idealized using the rider model. Table 7 shows the atomic coordinates (x10 4 ) and equivalent isotropic displacement parameters U(eq) is defined as one third of the trace of the orthogonalized Uij tensor. Table 8 shows the bond lengths and bond angle [°]. Table 9 shows the anisotropic displacement parameters .

[0373] result : This analysis confirmed the structure of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide. The asymmetric unit contains two molecules of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, two bromines (to balance the charge) and one methanol solvent molecule, as FIG. 16A to FIG. 16B The mirror image isomer setting was based on a Flack parameter refined to 0.038 (6). This study determined the absolute conformations at the chiral centers C15=S- and C35=S-.

[0374] Table 7.

[0375]

[0376]

[0377] Table 8.

[0378]

[0379]

[0380]

[0381]

[0382] Table 9.

[0383]

[0384]

[0385] Example A. THP-1 RPS6 ELISA assay

[0386] To measure phosphorylated ribosomal protein S6 (RPS6) in cell lysates, THP-1 cells (human acute monocytic leukemia) were purchased from ATCC (Manassas, VA) and maintained in RPMI (Gibco / Life Technologies, Carlsbad, CA) containing 10% FBS. For the assay, THP-1 cells were serum starved overnight in RPMI and then incubated in RPMI (2×10 5 Cells / well, 90 μL) were seeded into 96-well flat-bottom tissue culture treated plates (Corning, Corning, NY). Incubate at 37°C, 5% CO 2 The covered plates were incubated for 2 h at 37°C, 5% CO 2 The cells were lysed in a lysis buffer (Cell Signaling, Danvers, MA) containing protease inhibitors (Calbiochem / EMD, Germany), PMSF (Sigma, St Louis MO), HALTS (ThermoFisher, Rockford, IL) for 30 minutes. The cell lysates were frozen at -80 °C before testing. The lysates were tested in human / mouse / rat Phospho-RPS6 ELISA (R&D Systems, Inc., Minn, MN). The plates were measured using a microplate reader (SpectraMax M5–Molecular Devices, LLC Sunnyvale, CA) set to 450 nm, with a wavelength correction of 540. IC was calculated by fitting a curve of inhibitor percentage inhibition versus the logarithm of inhibitor concentration using GraphPad Prism 5.0 software. 50 Sure.

[0387] Example B. PI3K-γ Scintillation Proximity Assay

[0388] Material

[0389] [γ- 33P]ATP (10 mCi / mL) and wheat germ agglutinin (WGA) YSi SPA scintillation beads. The lipid kinase substrate 4,5-bisphosphate D-myo-phosphatidylinositol (3-O-phosphate-linked PtdIns (4,5) P2) D(+)-sn-1,2-di-O-octanoylglycerol (PIP2), CAS 204858-53-7 was purchased from Echelon Biosciences (Salt Lake City, UT). PI3Kγ (p110γ) recombinant human protein was purchased from Life technology (Grand Island, NY). ATP, MgCl were purchased from SigmaAldrich (St. Louis, MO). 2 , DTT, EDTA, MOPS and CHAPS.

[0390] Kinase reactions were performed in a polystyrene 384-well Greiner Bio-one white plate from Thermo Fisher Scientific in a final volume of 25 μL. The inhibitor was first serially diluted in DMSO and added to the wells of the plate, followed by the addition of the other reaction components. The final concentration of DMSO in the assay was 2%. The assay was carried out in 20 mM MOPS pH 6.7, 10 mM MgCl 2 PI3Kγ assay was performed in 5 mM DTT and CHAPS 0.03%. The reaction was initiated by adding ATP, and the final reaction mixture consisted of 20 μM PIP2, 2 μM ATP, 0.5 μCi [γ- 33 The reaction was incubated for 120 minutes and terminated by the addition of 40 μL of SPA beads suspended in the following quenching buffer: 163 mM potassium phosphate pH 7.8, 20% glycerol, 25 mM EDTA. The final concentration of SPA beads was 1.0 mg / mL. After the plates were sealed, the plates were shaken overnight at room temperature and centrifuged at 1500 rpm for 10 minutes, and the radioactivity of the products was determined by scintillation counting on a Topcount (Perkin-Elmer). IC was performed by fitting a curve of percent solvent control activity versus the logarithm of inhibitor concentration using GraphPad Prism 6.0 software. 50 Sure.

[0391] Example C. PI3Kδ Scintillation Proximity Assay

[0392] Material

[0393] [γ- 33P]ATP (10 mCi / mL) and wheat germ agglutinin (WGA) YSi SPA scintillation beads. The lipid kinase substrate 4,5-bisphosphate D-myo-phosphatidylinositol (3-O-phosphate-linked PtdIns (4,5) P2) D(+)-sn-1,2-di-O-octanoylglycerol (PIP2), CAS 204858-53-7 was purchased from Echelon Biosciences (Salt Lake City, UT). PI3Kδ (p110δ / p85α) recombinant human protein was purchased from Eurofins (St Charles, MO). ATP, MgCl were purchased from Sigma Aldrich (St. Louis, MO). 2 , DTT, EDTA, MOPS and CHAPS.

[0394] Kinase reactions were performed in a polystyrene 384-well Greiner Bio-one white plate from Thermo Fisher Scientific in a final volume of 25 μL. The inhibitor was first serially diluted in DMSO and added to the wells of the plate, followed by the addition of the other reaction components. The final concentration of DMSO in the assay was 2%. The plates were incubated at room temperature in 20 mM MOPS pH 6.7, 10 mM MgCl 2 PI3Kδ assay was performed in 5 mM DTT and CHAPS 0.03%. The reaction was initiated by adding ATP, and the final reaction mixture consisted of 20 μM PIP2, 2 μM ATP, 0.5 μCi [γ- 33 The reaction was incubated for 120 min and terminated by the addition of 40 μL of SPA beads suspended in the following quenching buffer: 163 mM potassium phosphate pH 7.8, 20% glycerol, 25 mM EDTA. The final concentration of SPA beads was 1.0 mg / mL. After the plates were sealed, the plates were shaken overnight at room temperature and centrifuged at 1500 rpm for 10 min, and the radioactivity of the products was determined by scintillation counting on a Topcount (PerkinElmer). IC was performed by fitting a curve of percent solvent control activity versus the logarithm of inhibitor concentration using GraphPad Prism 6.0 software. 50 Sure.

[0395] The compounds of Examples 1, 5, 6 and 9 were tested in the assays described in Examples A, B and C and were found to have the IC values ​​shown in Table A below. 50 value.

[0396] Table A.IC 50 value

[0397]

[0398] + refers to IC 50 ≤100nM; ++ refers to IC 50 ≤500nM; +++ refers to IC 50 <2000nM; ++++ refers to IC 50 ≥2000nM.

[0399] # refers to IC 50 ≤100nM; ## refers to IC 50 ≤500nM; ### refers to IC 50 <1000nM; #### refers to IC 50 ≥1000nM.

[0400] Based on the previous description, various modifications to the present invention in addition to those described herein will also be apparent to those skilled in the art. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications and disclosures, is incorporated herein by reference in its entirety.

Claims

1. A crystalline form of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionamide, wherein the crystalline form is selected from: Form IA having an X-ray powder diffraction pattern comprising the following peaks, expressed in 2θ: 10.3°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 19.2°±0.2°; and 24.0°±0.2°; Form IIA having an X-ray powder diffraction pattern comprising the following peaks, expressed in 2θ: 12.6°±0.2°; 16.9°±0.2°; 19.0°±0.2°; 20.1°±0.2°; 20.5°±0.2°; and 21.9°±0.2°; and Form IIIA having an X-ray powder diffraction pattern comprising the following peaks, expressed in 2θ: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; 17.9°±0.2°; and 20.3°±0.2°.

2. The crystalline form of claim 1 which is anhydrous and non-solvated.

3. The crystalline form of claim 1 or 2, which is Form IA.

4. A crystalline form as claimed in claim 3, which has an X-ray powder diffraction pattern substantially as shown in Figure 1.

5. The crystalline form of claim 3, having a DSC thermogram comprising an endothermic peak having a maximum at about 193°C.

6. The crystalline form of claim 3, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 2.

7. The crystalline form of claim 3, having a thermogravimetric analysis (TGA) substantially as shown in Figure 3.

8. The crystalline form of claim 1 or 2, which is Form IIA.

9. A crystalline form as claimed in claim 8, having an X-ray powder diffraction pattern substantially as shown in Figure 4.

10. The crystalline form of claim 8, having a DSC thermogram comprising an endothermic peak having a maximum at about 180°C.

11. The crystalline form of claim 8, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 5.

12. The crystalline form of claim 8, having a thermogravimetric analysis (TGA) substantially as shown in Figure 6.

13. The crystalline form of claim 1 or 2, which is Form IIIA.

14. A crystalline form according to claim 13, having an X-ray powder diffraction pattern substantially as shown in Figure 7.

15. The crystalline form of claim 13, having a DSC thermogram comprising an endothermic peak having a maximum at about 143°C.

16. The crystalline form of claim 13, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 8.

17. The crystalline form of claim 13, having a thermogravimetric analysis (TGA) substantially as shown in FIG.

9.

18. A crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, wherein the crystalline form is selected from: Form IB having an X-ray powder diffraction pattern comprising the following peaks, expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°; and Form IIB having an X-ray powder diffraction pattern comprising the following peaks, expressed in 2θ: 4.3°±0.2°; 7.4°±0.2°; 15.3°±0.2°; 17.0°±0.2°; 18.8°±0.2°; and 20.1°±0.2°.

19. The crystalline form of claim 18, which is anhydrous and non-solvated.

20. The crystalline form of claim 18 or 19, which is Form IB.

21. The crystalline form of claim 20, having an X-ray powder diffraction pattern substantially as shown in Figure 10.

22. The crystalline form of claim 20, having a DSC thermogram comprising an endothermic peak having a maximum at about 174°C.

23. The crystalline form of claim 20, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 11.

24. The crystalline form of claim 18 or 19, which is Form IIB.

25. The crystalline form of claim 24, having an X-ray powder diffraction pattern substantially as shown in Figure 12.

26. The crystalline form of claim 24, having a DSC thermogram comprising an endothermic peak having a maximum at about 165°C.

27. The crystalline form of claim 24, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 13.

28. A crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxyprop-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, wherein the crystalline form is Form IC having an X-ray powder diffraction pattern comprising the following peaks expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; 21.2°±0.2° and 24.8°±0.2°.

29. The crystalline form of claim 28, which is anhydrous and non-solvated.

30. The crystalline form of claim 28 or 29, which has an X-ray powder diffraction pattern substantially as shown in Figure 14.

31. The crystalline form of claim 28 or 29, having a DSC thermogram comprising an endothermic peak with a maximum at about 179°C.

32. The crystalline form of claim 28 or 29, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 15.

33. A salt which is 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide.

34. The salt of claim 33, having a 1:1 stoichiometric ratio of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide to hydrobromic acid.

35. The salt of claim 33 or 34 which is substantially isolated.

36. The salt of claim 33 or 34, which is in crystalline form.

37. The salt of claim 36, wherein the crystalline form is a solvated crystalline form.

38. The salt of claim 37, wherein the crystalline form is a methanol solvate crystalline form.

39. A composition comprising the crystalline form of any one of claims 1, 18 or 28 or the salt of claim 33.

40. The composition of claim 39, wherein the composition further comprises at least one pharmaceutically acceptable carrier.

41. Use of the crystalline form of any one of claims 1, 18 or 28 or the salt of claim 33 for the preparation of a medicament for inhibiting the activity of PI3K gamma kinase.

42. The use of claim 41, wherein the crystalline form is an inhibitor that is selective for PI3Kγ over one or more of PI3Kα, PI3Kβ and PI3Kδ.

43. Use of the crystalline form of any one of claims 1, 18 or 28 or the salt of claim 33 in the preparation of a medicament for treating a disease or condition in a patient, wherein the disease or condition is associated with abnormal expression or activity of PI3Kγ kinase.

44. The use of claim 43, wherein the disease or disorder is an autoimmune disease or disorder, cancer, cardiovascular disease, or a neurodegenerative disease.

45. The use of claim 43, wherein the disease or condition is lung cancer, melanoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, colon cancer, endometrial cancer, bladder cancer, skin cancer, uterine cancer, kidney cancer, gastric cancer, seminoma, teratoma, astrocytoma, neuroblastoma, glioma or sarcoma.

46. ​​The method of claim 45, wherein the sarcoma is an Askin's tumor, a botryoid sarcoma, a chondrosarcoma, an Ewing's sarcoma, a malignant hemangioendothelioma, a malignant schwannoma, an osteosarcoma, an alveolar soft part sarcoma, angiosarcoma, cystic sarcoma phyllodes, a dermatofibrosarcoma protuberans, a desmoid tumor, a desmoid small round cell tumor, an epithelioid sarcoma, a fibrosarcoma, a gastrointestinal stromal tumor (GIST), an hemangiopericytoma, angioendothelioma, a Kaposi's sarcoma, a leiomyosarcoma, a liposarcoma, a lymphangiosarcoma, a lymphosarcoma, a malignant peripheral nerve sheath tumor (MPNST), a neurofibrosarcoma, a rhabdomyosarcoma, a synovial sarcoma, or an undifferentiated pleomorphic sarcoma.

47. The use according to claim 46, wherein the osteosarcoma is extraskeletal chondrosarcoma or extraskeletal osteosarcoma.

48. The method of claim 43, wherein the disease or disorder is acute myeloid leukemia, acute monocytic leukemia, small lymphocytic lymphoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma, T-cell acute lymphoblastic leukemia (T-ALL), cutaneous T-cell lymphoma, large granular lymphocytic leukemia, mature peripheral T-cell neoplasm (PTCL), anaplastic large cell lymphoma (ALCL), or lymphoblastic lymphoma.

49. The method of claim 48, wherein the mature peripheral T-cell neoplasm (PTCL) is T-cell prolymphocytic leukemia, T-cell granular lymphocytic leukemia, aggressive NK-cell leukemia, mycosis fungoides / Sezary syndrome, T-cell anaplastic large cell lymphoma, enteropathy-type T-cell lymphoma, adult T-cell leukemia / lymphoma or angioimmunoblastic T-cell lymphoma.

50. The use according to claim 49, wherein the T-cell anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL.

51. The use of claim 43, wherein the disease or condition is Burkitt's lymphoma, acute myeloblastic leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanthoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenstrom's macroglobulinemia, prolymphoma Myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, latent myeloma, monoclonal gammopathy of undetermined significance (MGUS), or diffuse large B-cell lymphoma.

52. The use of claim 51, wherein the non-Hodgkin's lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, indolent NHL (iNHL) or aggressive NHL (aNHL).

53. The use of claim 51, wherein the diffuse large B-cell lymphoma is activated B-cell-like (ABC) diffuse large B-cell lymphoma or germinal center B-cell (GCB) diffuse large B-cell lymphoma.

54. The use of claim 51, wherein the Burkitt's lymphoma is endemic Burkitt's lymphoma, sporadic Burkitt's lymphoma, or Burkitt's lymphoma-like.

55. The method of claim 43, wherein the disease or disorder is rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, asthma, allergic reaction, pancreatitis, psoriasis, glomerulonephritis, inflammatory bowel disease, thrombosis, meningitis, encephalitis, diabetic retinopathy, benign prostatic hypertrophy, myasthenia gravis, Sjogren's syndrome, osteoarthritis, restenosis, or atherosclerosis.

56. The use according to claim 55, wherein the allergic reaction is allergic rhinitis or anaphylaxis.

57. The method of claim 43, wherein the disease or disorder is cardiac hypertrophy, myocardial cell dysfunction, acute coronary syndrome, chronic obstructive pulmonary disease (COPD), chronic bronchitis, elevated blood pressure, ischemia, ischemia-reperfusion, vasoconstriction, anemia, bacterial infection, viral infection, transplant rejection, nephropathy, anaphylactic shock fibrosis, skeletal muscle atrophy, skeletal muscle hypertrophy, angiogenesis, sepsis, graft-versus-host disease, allogeneic or xenotransplantation, glomerulosclerosis, progressive renal fibrosis, idiopathic thrombocytopenic purpura (ITP), autoimmune hemolytic anemia, vasculitis, systemic lupus erythematosus, or pemphigus.

58. The use of claim 57, wherein the renal disease is lupus nephritis or membranous nephropathy.

59. The use of claim 57, wherein the idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP.

60. The method of claim 57, wherein the vasculitis is Behcet's disease, Cogan's syndrome, giant cell arteritis, polymyalgia rheumatica (PMR), Takayasu's arteritis, Buerger's disease, central nervous system vasculitis, Kawasaki disease, polyarteritis nodosa, Tsagaer-Strauss syndrome, primary or hepatitis C virus (HCV)-induced mixed cryoglobulinemia vasculitis, Henoch-Schwann purpura (HSP), allergic vasculitis, microscopic polyangiitis, Wegener's granulomatosis, or anti-neutrophil cytoplasmic antibody-associated (ANCA) systemic vasculitis (AASV).

61. The use of claim 43, wherein the disease or disorder is Alzheimer's disease, central nervous system trauma, or stroke.

62. A method of preparing a crystalline form of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, wherein the crystalline form is Form IA having an X-ray powder diffraction pattern comprising the following peaks, expressed in 2θ: 10.3°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 19.2°±0.2°; and 24.0°±0.2°, wherein the method comprises: A vial was charged with the compound as a solid and stirred at 80 °C while a 1:2 solution of isopropyl acetate:heptane was added dropwise; allowing the solid compound to dissolve while forming a second solid on the wall of the vial, with stirring at 80° C. for about 2 hours; Cooling the vial to ambient temperature; filtering the second solid and washing with heptane; as well as The second solid was collected and dried under reduced pressure for about 30 minutes to afford Form IA of the compound as a white solid.

63. A method of preparing a crystalline form of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, wherein the crystalline form is Form IIA having an X-ray powder diffraction pattern comprising the following peaks, expressed in 2θ: 12.6°±0.2°; 16.9°±0.2°; 19.0°±0.2°; 20.1°±0.2°; 20.5°±0.2°; and 21.9°±0.2°, wherein the method comprises: The free base of the compound was dissolved in isopropyl acetate in a vial to form a solution; With stirring at ambient temperature, heptane was added to the solution in a ratio of 1:2 isopropyl acetate:heptane; The solution was heated at 80°C with stirring for about 2 hours; allowing the mixture to cool to ambient temperature and stirring for about 1 hour, whereupon a solid formed; and The solid was collected by filtration and air dried to provide Form IIA of the compound.

64. A method of preparing a crystalline form of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, wherein the crystalline form is Form IIIA having an X-ray powder diffraction pattern comprising the following peaks expressed in 2θ: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; 17.9°±0.2°; and 20.3°±0.2°, wherein the method comprises dissolving the compound in a solvent to form a mixture, and crystallizing the compound from the mixture, wherein the solvent comprises methanol, wherein the method further comprises: heating the mixture to a temperature of 50°C to 70°C; as well as The mixture was allowed to cool to room temperature.

65. A method of preparing a crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, wherein the crystalline form is Form IB having an X-ray powder diffraction pattern comprising the following peaks expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°, the method comprising dissolving the compound in a solvent to form a mixture, and crystallizing the compound from the mixture, the solvent comprising isopropyl acetate, wherein the method further comprises: heating the mixture to a temperature of 70°C to 90°C, and The mixture was allowed to cool to room temperature.

66. A method of preparing a crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, wherein the crystalline form is Form IIB having an X-ray powder diffraction pattern comprising the following peaks expressed in 2θ: 4.3°±0.2°; 7.4°±0.2°; 15.3°±0.2°; 17.0°±0.2°; 18.8°±0.2°; and 20.1°±0.2°, the method comprising dissolving the compound in a solvent to form a mixture, and crystallizing the compound from the mixture, the solvent comprising isopropyl acetate and heptane, wherein the method further comprises: heating the mixture to a temperature of 70°C to 90°C, and The mixture was allowed to cool to room temperature.

67. A method of preparing a crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, wherein the crystalline form is Form IC having an X-ray powder diffraction pattern comprising the following peaks expressed in 2θ: 6.2°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; 21.2°±0.2° and 24.8°±0.2°, the method comprising dissolving the compound in a solvent to form a mixture, and crystallizing the compound from the mixture, the solvent comprising isopropyl acetate and heptane, wherein the method further comprises: heating the mixture to a temperature of 70°C to 90°C, and The mixture was allowed to cool to room temperature.

68. A method for preparing a hydrobromide salt of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropionamide, the method comprising dissolving the compound in a solvent to form a mixture, and adding hydrobromic acid to the mixture, the solvent comprising methanol, wherein: The hydrobromic acid is added to the mixture in the form of an aqueous solution of hydrobromic acid, and To the mixture was added 1.1 to 1.5 equivalents of hydrobromic acid based on 1 equivalent of the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

69. The method of claim 68, further comprising substantially isolating the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide salt.

70. The method of claim 69, wherein the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide salt is isolated in a crystalline form.

71. The method of claim 69, wherein the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide salt is isolated as a methanol solvate crystalline form.

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