Crystalline forms of phosphoinositide 3-kinase (PI3k) inhibitor

Abstract

To provide crystalline forms of PI3K inhibitors useful in the treatment of cancer and other diseases.SOLUTION: The present invention relates to crystalline forms of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to salts and crystalline forms of the PI3K inhibitors 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, 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 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, which are useful in the treatment of cancer and other diseases. BACKGROUND OF THE INVENTION

[0002] The compounds 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide having formula A, 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, and 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 having formula C. [ka] are phosphoinositide 3-kinase (PI3K) inhibitors useful in the treatment of various diseases, including cancer. For drug development, it is generally advantageous to utilize forms of the drug that have desirable properties with respect to preparation, purification, reproducibility, stability, bioavailability, and other characteristics. Thus, the salt and crystalline forms of the compounds of Formula A, Formula B, and Formula C provided herein address the continuing need for the development of PI3K inhibitors for the treatment of serious diseases. Summary of the Invention

[0003] The present invention relates to 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: [ka] to provide.

[0004] The present invention further provides compositions comprising salts and crystalline forms of the compound of formula A as set forth herein and at least one pharmaceutically acceptable carrier. The present invention further provides compositions comprising a crystalline form of the compound of formula B and at least one pharmaceutically acceptable carrier. The present invention further provides compositions comprising a crystalline form of the compound of formula C and at least one pharmaceutically acceptable carrier.

[0005] The present invention further provides processes for preparing the salts and crystalline forms of the present invention.

[0006] The present invention further provides a method of treating a disease associated with aberrant expression or activity of PI3K kinase in a patient, comprising administering to the patient a therapeutically effective amount of a salt or crystalline form of the invention. [Brief explanation of the drawings]

[0007] [Figure 1]1 shows the XRPD pattern for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IA. [Figure 2] 1 shows the results of a DSC experiment for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IA. [Figure 3] 1 shows the results of a TGA experiment for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IA. [Figure 4] FIG. 1 shows the XRPD pattern for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIA. [Figure 5] 1 shows the results of a DSC experiment for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIA. [Figure 6] 1 shows the results of a TGA experiment for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIA. [Figure 7] 1 shows the XRPD pattern for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIIA. [Figure 8]1 shows the results of a DSC experiment on 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIIA. [Figure 9] 1 shows the results of a TGA experiment on 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIIA. [Figure 10] 1 shows the XRPD pattern for 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. [Figure 11] 1 shows the results of a DSC experiment on 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. [Figure 12] Figure 1 shows the XRPD pattern for 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. [Figure 13] 1 shows the results of a DSC experiment on 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. [Figure 14] Figure 1 shows the XRPD pattern for 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. [Figure 15] 1 shows the results of a DSC experiment on 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. [Figure 16A] Thermal ellipsoids drawn at a 30% probability level are used to depict the 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 form. [Figure 16B] Crystalline units of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide are shown using thermal ellipsoids drawn at a 30% probability level. DETAILED DESCRIPTION OF THE INVENTION

[0008] The present invention relates inter alia 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: [ka] This is useful, for example, in preparing solid dosage forms of the compounds for the treatment of various diseases, including cancer.

[0009] The present invention also relates, inter alia, 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: [ka]

[0010] The present invention also relates, inter alia, to a crystalline form of the PI3K inhibitor 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, having formula C: [ka]

[0011] Generally, different crystalline forms of the same substance have different bulk properties, e.g., with respect to hygroscopicity, solubility, stability, etc. Forms with high melting points often have good thermodynamic stability, which is advantageous for extending the shelf life of solid-state drug formulations. Forms with low melting points are often less thermodynamically stable, but are advantageous in that they result in increased water solubility and therefore higher drug bioavailability. Forms with low hygroscopicity are desirable for their stability to heat and humidity and resist degradation during long-term storage. Anhydrous forms are often desirable because they can be consistently produced without worrying about changes in weight or composition due to changes in solvent or water content. On the other hand, hydrated or solvated forms may be advantageous in that they are less likely to be hygroscopic and may exhibit improved stability to humidity under storage conditions.

[0012] As used herein, the term "crystalline form" refers to a particular lattice configuration of a crystalline substance. Different crystalline forms of the same substance generally have different crystal lattices (e.g., unit cells), which are due to the different physical properties unique to each crystalline form. In some cases, different lattice configurations have different water or solvent contents. Different crystalline lattices can be identified by solid-state characterization methods, such as X-ray powder diffraction (XRPD). Other characterization methods, such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), solid-state NMR, etc., further aid in the identification of crystalline forms, as well as in determining stability and solvent / water content.

[0013] Crystalline forms of a substance include both solvated (e.g., hydrated) and nonsolvated (e.g., anhydrous) forms. Hydrated forms are crystalline forms that contain water in the crystal lattice. Hydrated forms may be stoichiometric hydrates, in which water is present in the lattice at a specific water / molecule ratio, such as hemihydrate, monohydrate, or dihydrate. Hydrated forms may also be nonstoichiometric, in which the water content is variable and dependent on external conditions such as humidity.

[0014] Crystalline forms are most commonly characterized by XRPD. The XRPD pattern of reflections (peaks) is typically considered a fingerprint of a particular crystalline form. It is well known that the relative intensities of XRPD peaks can vary widely depending on, among other things, sample preparation techniques, crystal size distribution, filters, sample mounting procedures, and the particular instrument used. In some cases, new peaks may be observed or existing peaks may disappear depending on the type or settings of the instrument (e.g., whether a Ni filter is used). As used herein, the term "peak" refers to a reflection having a relative height / intensity of at least about 4% of the maximum peak height / intensity. Furthermore, instrumental variations and other factors can affect 2-theta values. Thus, peak assignments such as those reported herein may vary by plus or minus about 0.2° (2-theta), and the term "substantially," as used herein in the context of XRPD, is meant to encompass such variations.

[0015] Similarly, temperature measurements for DSC, TGA, or other thermal experiments can vary by approximately ±4°C depending on the instrument, the particular settings, sample preparation, etc. For example, with DSC, it is known that the observed temperatures depend on the rate of temperature change as well as the sample preparation technique and particular instrument utilized. Thus, values ​​reported herein for DSC thermograms, as noted above, can vary by ±4°C. Thus, it is understood that crystalline forms reported herein having DSC thermograms "substantially" as shown in any figure accommodate such variations.

[0016] Crystalline Form of the Compound of Formula A The compound of Formula A may be isolated in multiple crystalline forms, including, for example, crystalline forms that are anhydrous and / or unsolvated or solvated. 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 unsolvated. In some embodiments, the crystalline form of the compound of Formula A is anhydrous and unsolvated. "Anhydrous" means that the crystalline form of the compound of Formula A is essentially free of bound water in the crystal lattice structure, i.e., the compound does not form crystalline hydrates.

[0017] In some embodiments, the present application provides a process 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 process 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.

[0018] In some embodiments, the solvent comprises isopropyl acetate. In some embodiments, the solvent further comprises heptane.

[0019] In some embodiments, the solvent comprises methanol.

[0020] In some embodiments, the process further comprises heating the mixture to a temperature of about 70°C to about 90°C.

[0021] In some embodiments, the process further comprises heating the mixture to a temperature of about 50°C to about 70°C.

[0022] In some embodiments, the process further comprises cooling the mixture to room temperature.

[0023] The present application further provides crystalline forms of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide prepared according to the processes set forth herein. In some embodiments, the crystalline form is Form IA described herein. In some embodiments, the crystalline form is Form IIA described herein. In some embodiments, the crystalline form is Form IIIA described herein.

[0024] In some embodiments, the present application provides a process for preparing the 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-hydroxypropanamide. In some embodiments, the process includes dissolving a compound of Formula A in a solvent to form a mixture and adding hydrobromic acid to the mixture.

[0025] In some embodiments of the process for preparing the hydrobromide salt, the solvent comprises methanol.

[0026] In some embodiments of the process for preparing the hydrobromide salt, hydrobromic acid is added to the mixture as an aqueous solution of hydrobromic acid.

[0027] In some embodiments of the process for preparing the hydrobromide salt, an excess amount of hydrobromic acid 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 is added to the mixture.

[0028] In some embodiments of the process 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.

[0029] In some embodiments of the process for preparing a hydrobromide salt, the process further comprises substantially isolating 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide.

[0030] In some embodiments of the process for preparing a hydrobromide salt, 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide is isolated as a crystalline form.

[0031] In some embodiments of the process for preparing the hydrobromide salt, 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide is isolated as a methanol solvate crystalline form.

[0032] In some embodiments, the present application further provides the hydrobromide salt of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, prepared according to the processes set forth herein. In some embodiments, the hydrobromide salt is crystalline. In some embodiments, the hydrobromide salt is in a solvated crystalline form. In some embodiments, the hydrobromide salt is in a methanol solvated crystalline form.

[0033] In some embodiments, the crystalline forms of the present invention are substantially isolated. By "substantially isolated," it is meant that a particular crystalline form of the compound of Formula A is at least partially isolated 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 those that are not substantially isolated crystalline forms, including, for example, other crystalline forms and other substances.

[0034] 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 contains 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.

[0035] Crystalline Form IA of the compound of formula A In some embodiments, the crystalline form of the compound of formula A is Form IA. In some embodiments, crystalline Form IA of the compound of formula A is anhydrous and ansolvated. The preparation of Form IA of the compound of formula A is described in Example 1. Crystalline Form IA of the compound of formula A can be prepared generally as described in Example 2.

[0036] Crystalline Form IA of the compound of Formula A can be identified by unique signatures for, for example, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, crystalline Form IA of the compound of Formula A is characterized by an XRPD pattern substantially as shown in Figure 1. The peaks of the XRPD pattern are listed in Table 1.

[0037] In some embodiments, crystalline form IA of the compound of formula A is characterized by an XRPD pattern comprising a peak at 8.6°±0.2° in term of 2θ. In some embodiments, crystalline form IA of the compound of formula A is characterized by an XRPD pattern comprising a peak at 9.5°±0.2° in term of 2θ. In some embodiments, crystalline form IA of the compound of formula A is characterized by an XRPD pattern comprising a peak at 10.3°±0.2° in term of 2θ. In some embodiments, crystalline form IA of the compound of formula A is characterized by an XRPD pattern comprising a peak at 14.9°±0.2° in term of 2θ.

[0038] In some embodiments, crystalline form IA of the compound of formula A has an XRPD pattern comprising the following peaks 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 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 in 2θ: 10.3°±0.2°; 14.9°±0.2°; 17.3°±0.2°; 19.2°±0.2°; and 24.0°±0.2°.

[0039] In some embodiments, crystalline form IA 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 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 two or more, three or more, or four or more of the following peaks 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 two or more, three or more, or four or more of the following peaks 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°.

[0040] 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, crystalline Form IA of the compound of Formula A has a DSC thermogram substantially as shown in FIG.

[0041] In some embodiments, crystalline Form IA of the compound of formula A has a TGA trace substantially as shown in FIG.

[0042] Crystalline Form IIA of the compound of formula A In some embodiments, the crystalline form of the compound of Formula A is Form IIA. In some embodiments, crystalline Form IIA of the compound of Formula A is anhydrous and ansolvated. This crystalline form can be prepared generally as described in Example 3.

[0043] Crystalline Form IIA of the compound of Formula A can be identified by unique signatures for, for example, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern substantially as shown in Figure 4. The peaks of the XRPD pattern are listed in Table 2.

[0044] In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern including a peak at 9.1°±0.2° in term of 2θ. In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern including a peak at 11.1°±0.2° in term of 2θ. In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern including a peak at 21.9°±0.2° in term of 2θ. In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern including a peak at 12.6°±0.2° in term of 2θ. In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern including a peak at 13.6°±0.2° in term of 2θ. In some embodiments, crystalline Form IIA of the compound of Formula A is characterized by an XRPD pattern including a peak at 18.0°±0.2° in term of 2θ. In some embodiments, crystalline Form IIA of the compound of formula A is characterized by an XRPD pattern comprising a peak at 19.0°±0.2° in terms of 2θ.

[0045] In some embodiments, crystalline Form IIA of the compound of formula A has an XRPD pattern comprising the following peaks 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 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 in 2θ: 12.6°±0.2°; 18.0°±0.2°; 19.0°±0.2°; and 21.9°±0.2°.

[0046] In some embodiments, 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 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, 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 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 two or more, three or more, or four or more of the following peaks in 2θ: 12.6°±0.2°; 18.0°±0.2°; 19.0°±0.2°; 20.5°±0.2°; and 21.9°±0.2°.

[0047] In some embodiments, Form IIA of the compound of Formula A is characterized by a DSC thermogram comprising an endothermic peak having a maximum at about 180° C. In some embodiments, crystalline Form IIA of the compound of Formula A has a DSC thermogram substantially as shown in FIG.

[0048] In some embodiments, crystalline Form IIA of the compound of formula A has a TGA trace substantially as shown in FIG.

[0049] Crystalline Form IIIA of the compound of formula A In some embodiments, the crystalline form of the compound of Formula A is Form IIIA. In some embodiments, crystalline Form IIIA of the compound of Formula A is anhydrous and ansolvated. This crystalline form can be prepared generally as described in Example 4.

[0050] Crystalline Form IIIA of the compound of Formula A can be identified by unique signatures for, for example, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, crystalline Form IIIA of the compound of Formula A is characterized by an XRPD pattern substantially as shown in Figure 7. The peaks of the XRPD pattern are listed in Table 3.

[0051] In some embodiments, crystalline Form IIIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak at 8.1°±0.2° in term of 2θ. In some embodiments, crystalline Form IIIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak at 10.6°±0.2° in term of 2θ. In some embodiments, crystalline Form IIIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak at 13.5°±0.2° in term of 2θ. In some embodiments, crystalline Form IIIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak at 14.2°±0.2° in term of 2θ. In some embodiments, crystalline Form IIIA of the compound of Formula A is characterized by an XRPD pattern comprising a peak at 20.3°±0.2° in term of 2θ.

[0052] In some embodiments, crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising the following peaks in 2θ: 8.1°±0.2°; 10.6°±0.2°; 13.5°±0.2°; and 14.2°±0.2°. In some embodiments, crystalline Form IIIA of the compound of Formula A has an XRPD pattern comprising the following peaks 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 in 2θ: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; and 20.3°±0.2°.

[0053] In some embodiments, crystalline Form IIIA 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 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 two or more, three or more, or four or more of the following peaks 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 two or more, three or more, or four or more of the following peaks 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°.

[0054] 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, crystalline Form IIIA of the compound of Formula A has a DSC thermogram substantially as shown in FIG.

[0055] In some embodiments, crystalline Form IIIA of the compound of formula A has a TGA trace substantially as shown in FIG.

[0056] Hydrobromide of Formula A 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 is 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide in a 1:1 stoichiometric ratio to hydrobromic acid. The hydrobromide salt form can be prepared generally as described in Example 11.

[0057] 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 in a solvated crystalline form. In some embodiments, the hydrobromide salt of the compound of Formula A is in a methanol solvate crystalline form.

[0058] Crystalline Form of the Compound of Formula B The compound of formula B may be isolated in multiple crystalline forms, including, for example, crystalline forms that are anhydrous and / or ansolvated. 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 ansolvated. In some embodiments, the crystalline form of the compound of formula B is anhydrous and ansolvated. "Anhydrous" means that the crystalline form of the compound of formula B is essentially free of bound water in the crystal lattice structure, i.e., the compound does not form crystalline hydrates.

[0059] In some embodiments, the crystalline forms provided herein can be prepared, for example, by a process 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.

[0060] In some embodiments, the process further comprises heating the mixture to a temperature of about 70° C. to about 90° C. In some embodiments, the process further comprises cooling the mixture to room temperature.

[0061] In some embodiments, the solvent comprises isopropyl acetate. In some embodiments, the solvent further comprises heptane.

[0062] 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, prepared according to the processes set forth 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.

[0063] In some embodiments, the crystalline forms of the present invention are substantially isolated. By "substantially isolated," it is meant that a particular crystalline form of the compound of Formula B is at least partially isolated 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 those that are not substantially isolated crystalline forms, including, for example, other crystalline forms and other substances.

[0064] 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 contains 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.

[0065] Crystalline Form IB of the compound of formula B In some embodiments, the crystalline form of the compound of formula B is form IB. In some embodiments, crystalline form IB of the compound of formula B is anhydrous and ansolvated. The preparation of the compound of formula B is described in Examples 5 and 6. Crystalline form IB can be prepared generally as described in Example 7.

[0066] Crystalline Form IB of the compound of Formula B can be identified, for example, by unique signatures for X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, crystalline Form IB of the compound of Formula B is characterized by an XRPD pattern substantially as shown in Figure 10. The peaks of the XRPD pattern are listed in Table 4.

[0067] In some embodiments, crystalline form IB of the compound of Formula B is characterized by an XRPD pattern comprising a peak at 6.2°±0.2° in term of 2θ. In some embodiments, crystalline form IB of the compound of Formula B is characterized by an XRPD pattern comprising a peak at 15.6°±0.2° in term of 2θ. In some embodiments, crystalline form IB of the compound of Formula B is characterized by an XRPD pattern comprising a peak at 20.7°±0.2° in term of 2θ. In some embodiments, crystalline form IB of the compound of Formula B has an XRPD pattern comprising the following peaks in term of 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 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 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 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°.

[0068] In some embodiments, crystalline form IB of the compound of formula B has an XRPD pattern comprising two or more, three or more, or four or more of the following peaks 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° 2θ. In some embodiments, crystalline form IB of the compound of formula B has an XRPD pattern comprising two or more, three or more, or four or more of the following peaks 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 two or more, three or more, or four or more of the following peaks 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, crystalline form IB of the compound of formula B has an XRPD pattern comprising two or more, three or more, or four or more of the following peaks 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°.

[0069] 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, crystalline Form IB of the compound of Formula B has a DSC thermogram substantially as shown in FIG.

[0070] Crystalline Form IIB of the compound of formula B In some embodiments, the crystalline form of the compound of Formula B is Form IIB. In some embodiments, crystalline Form IIB of the compound of Formula B is anhydrous and ansolvated. This crystalline form can be prepared generally as described in Example 8.

[0071] Crystalline Form IIB of the compound of Formula B can be identified, for example, by unique signatures for X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern substantially as shown in Figure 12. The peaks of the XRPD pattern are listed in Table 5.

[0072] In some embodiments, crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern including a peak at 4.2°±0.2° in term of 2θ. In some embodiments, crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern including a peak at 7.4°±0.2° in term of 2θ. In some embodiments, crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern including a peak at 13.3°±0.2° in term of 2θ. In some embodiments, crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern including a peak at 20.1°±0.2° in term of 2θ. In some embodiments, crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern including a peak at 17.0°±0.2° in term of 2θ. In some embodiments, crystalline Form IIB of the compound of Formula B is characterized by an XRPD pattern including a peak at 18.8°±0.2° in term of 2θ.

[0073] In some embodiments, crystalline Form IIB of the compound of Formula B has an XRPD pattern comprising the following peaks 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 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 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°.

[0074] In some embodiments, crystalline Form IIB of the compound of Formula B has an XRPD pattern comprising two or more, three or more, or four or more of the following peaks 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 two or more, three or more, or four or more of the following peaks 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°.

[0075] 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, crystalline Form IIB of the compound of Formula B has a DSC thermogram substantially as shown in FIG.

[0076] Crystalline Form of the Compound of Formula C The compound of formula C may be isolated in multiple crystalline forms, including, for example, crystalline forms that are anhydrous and / or ansolvated. 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 ansolvated. In some embodiments, the crystalline form of the compound of formula C is anhydrous and ansolvated. By "anhydrous" is meant that the crystalline form of the compound of formula C is essentially free of bound water in the crystal lattice structure, i.e., the compound does not form crystalline hydrates.

[0077] In some embodiments, the present application provides a process for 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. In some embodiments, the process includes dissolving 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 in a solvent to form a mixture, and crystallizing the compound from the mixture.

[0078] In some embodiments, the process further comprises heating the mixture to a temperature of about 70° C. to about 90° C. In some embodiments, the process further comprises cooling the mixture to room temperature.

[0079] In some embodiments, the solvent comprises isopropyl acetate. In some embodiments, the solvent further comprises heptane.

[0080] The present application provides 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 prepared according to the processes set forth herein. In some embodiments, the crystalline form is Form IC as described herein.

[0081] In some embodiments, the crystalline forms of the present invention are substantially isolated. By "substantially isolated," it is meant that a particular crystalline form of the compound of Formula C is at least partially isolated 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 those that are not substantially isolated crystalline forms, including, for example, other crystalline forms and other substances.

[0082] 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 contains 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.

[0083] Crystalline Form IC of the compound of formula C In some embodiments, the crystalline form of the compound of formula C is crystalline form IC. In some embodiments, crystalline form IC of the compound of formula C is anhydrous and ansolvated. The preparation of the compound of formula C is described in Example 9. Crystalline form IC of the compound of formula C can be prepared generally as described in Example 10.

[0084] Crystalline Form IC of the compound of Formula C can be identified, for example, by unique signatures for X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, crystalline Form IC of the compound of Formula C is characterized by an XRPD pattern substantially as shown in Figure 14. The peaks of the XRPD pattern are listed in Table 6.

[0085] In some embodiments, crystalline Form IC of the compound of Formula C is characterized by an XRPD pattern comprising a peak at 6.2°±0.2° in term of 2θ. In some embodiments, crystalline Form IC of the compound of Formula C is characterized by an XRPD pattern comprising a peak at 11.9°±0.2° in term of 2θ. In some embodiments, crystalline Form IC of the compound of Formula C is characterized by an XRPD pattern comprising a peak at 16.7°±0.2° in term of 2θ. In some embodiments, crystalline Form IC of the compound of Formula C has an XRPD pattern comprising the following peaks 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 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 in 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.3°±0.2°; 11.9°±0.2°; and 12.5°±0.2°.

[0086] In some embodiments, crystalline form IC of the compound of formula C has an XRPD pattern comprising two or more, three or more, or four or more of the following peaks 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 two or more, three or more, or four or more of the following peaks 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 two or more, three or more, or four or more of the following peaks 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°.

[0087] In some embodiments, Form IC of the compound of Formula C is characterized by a DSC thermogram comprising an endothermic peak having a maximum at about 179° C. In some embodiments, crystalline Form IC of the compound of Formula C has a DSC thermogram substantially as shown in FIG.

[0088] How to use The compounds of the invention described herein (e.g., salts and crystalline forms) inhibit the activity of PI3K gamma kinase. Accordingly, the salts and crystalline forms of the invention described herein can be used in methods of inhibiting PI3K gamma kinase by contacting the kinase. In some embodiments, the salts and crystalline forms of the invention can be used in methods of inhibiting the activity of PI3K gamma in an individual / patient in need thereof by administering an effective amount of a salt or crystalline form described herein. In some embodiments, the modulation is inhibition. In some embodiments, the contacting is in vivo. In some embodiments, the contacting is ex vivo. Advantageously, the crystalline forms described herein demonstrate superior efficacy and a favorable safety and toxicity profile in animal studies.

[0089] In some embodiments, the PI3Kγ comprises a mutation. The mutation may be a substitution of one amino acid with another amino acid or a deletion of one or more amino acids. In such embodiments, the mutation may be in the kinase domain of the PI3Kγ.

[0090] In some embodiments, the salt or crystalline form further inhibits PI3Kδ.

[0091] The salts and crystalline forms described herein can be selective. "Selective" means that the salts or crystalline forms bind to or inhibit PI3Kγ with greater affinity or potency, respectively, compared to at least one other kinase. In some embodiments, the salts and crystalline forms of the present disclosure are selective inhibitors of PI3Kγ over PI3Kδ, PI3Kα, and PI3Kβ. In some embodiments, the salts and crystalline forms of the present disclosure are selective inhibitors of PI3Kγ over PI3Kα and PI3Kβ. In some embodiments, the selectivity can be at least about 2-fold, 3-fold, 5-fold, 10-fold, or 20-fold over PI3Kδ, as measured by the assays described herein. In some embodiments, the selectivity can be tested at an ATP concentration of 2 μM for each enzyme. In some embodiments, the selectivity of the salts and crystalline forms of the present disclosure can be determined by cellular assays related to specific PI3K kinase activity.

[0092] Another aspect of the present disclosure relates to a method of treating a kinase PI3Kγ-related disease or disorder in an individual (e.g., a patient) by administering to the individual in need thereof a therapeutically effective amount or dose of one or more salts or crystalline forms of the present disclosure or a pharmaceutical composition thereof. A PI3Kγ-related disease or disorder can include any disease, disorder, or condition directly or indirectly linked to the expression or activity of PI3Kγ, including overexpression and / or abnormal activity levels.

[0093] In some embodiments, the disease or disorder is an autoimmune disease or disorder, cancer, a cardiovascular disease, or a neurodegenerative disease.

[0094] 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 tumor, botryoid sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft part sarcoma, angiosarcoma, cystosarcoma phyllodes, dermatofibrosarcoma protuberans, desmoid tumor, desmoplastic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, angiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma, or undifferentiated pleomorphic sarcoma.

[0095] In some embodiments, the disease or disorder is mesothelioma or adenocarcinoma. In some embodiments, the disease or disorder is mesothelioma. In some embodiments, the disease or disorder is adrenal carcinoma.

[0096] In some embodiments, the disease or disorder is acute myeloid leukemia (e.g., acute monocytic leukemia), small lymphocytic lymphoma, chronic lymphocytic leukemia (CLL), chronic myelogenous 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 / Sézary syndrome, naplastic large cell lymphoma (T-cell type), enteropathic T-cell lymphoma, adult T-cell leukemia / lymphoma, or angioimmunoblastic T-cell lymphoma. In some embodiments, the anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL.

[0097] 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, prolymphocytic leukemia, acute lymphoblastic leukemia, acute 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, prolymphocytic leukemia, acute lymphoblastic leukemia, Blastic 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, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), or diffuse large B-cell lymphoma.

[0098] MDSCs (myeloid-derived suppressor cells) are a heterogeneous group of immune cells from the myeloid lineage (a family of cells originating from bone marrow stem cells). MDSCs proliferate strongly as a result of altered hematopoiesis in pathological conditions, such as chronic infection and cancer. MDSCs are distinct from other myeloid cell types in that they possess potent immunosuppressive activity rather than immunostimulatory properties. Like other myeloid cells, MDSCs interact with and regulate the function of other types of immune cells, including T cells, dendritic cells, macrophages, and natural killer cells. In some embodiments, for example, compounds and the like described herein can be used in methods related to cancerous tissues (e.g., tumors) with high MDSC infiltration, including solid tumors with high basal levels of macrophage and / or MDSC infiltration.

[0099] In some embodiments, the disease or disorder is selected from the group consisting of Burkitt's lymphoma, acute myeloblastic leukemia, chronic myelogenous 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, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), or diffuse large B-cell lymphoma.

[0100] In some embodiments, the non-Hodgkin's lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, indolent NHL (iNHL), or aggressive NHL (aNHL).

[0101] 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.

[0102] In some embodiments, the Burkitt lymphoma is endemic Burkitt lymphoma, sporadic Burkitt lymphoma, or Burkitt-like lymphoma.

[0103] In some embodiments, the disease or disorder is rheumatoid arthritis, multiple sclerosis, systemic lupus erythematous, asthma, allergy (e.g., allergic rhinitis), pancreatitis, psoriasis, anaphylaxis, glomerulonephritis, inflammatory bowel disease (e.g., Crohn's disease and ulcerative colitis), thrombosis, meningitis, encephalitis, diabetic retinopathy, benign prostatic hyperplasia, myasthenia gravis, Sjogren's syndrome, osteoarthritis, restenosis, or atherosclerosis.

[0104] In some embodiments, the disease or disorder is cardiac hypertrophy, cardiac myocyte 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 true erythrocytic 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 xenotransplantation, glomerulosclerosis, progressive renal fibrosis, idiopathic thrombocytopenic purpura (ITP), idiopathic pulmonary fibrosis, autoimmune hemolytic anemia, vasculitis, lupus nephritis, pemphigus, or membranous nephropathy.

[0105] In some embodiments, the disease or disorder is cardiac hypertrophy, cardiac myocyte dysfunction, chronic obstructive pulmonary disease (COPD), elevated blood pressure, ischemia, ischemia-reperfusion, vasoconstriction, anemia (e.g., hemolytic anemia, aplastic anemia, or true erythrocytic 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.

[0106] In some embodiments, the disease or disorder is Alzheimer's disease, central nervous system injury, or stroke.

[0107] In some embodiments, the idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP.

[0108] In some embodiments, the vasculitis is Behçet's 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 (essential or hepatitis C virus (HCV)-induced), Henoch-Schönlein purpura (HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener's granulomatosis, or antineutrophil cytoplasmic antibody-associated (ANCA) systemic vasculitis (AASV).

[0109] The present disclosure further provides a salt or crystalline form described herein for use in any of the methods described herein.

[0110] The present disclosure further 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.

[0111] 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 present disclosure includes administering a crystalline form of the present disclosure to an individual or patient, such as a human, who has PI3K, and also includes, for example, introducing a salt or crystalline form of the present disclosure into a sample containing a cell preparation or purified preparation containing PI3K.

[0112] It is believed that the salts and crystalline forms provided herein, or any of their embodiments, may have a satisfactory pharmacological profile and promising biopharmaceutical properties, such as toxicological profile, metabolic and pharmacokinetic properties, solubility, and permeability. It is understood that determining appropriate biopharmaceutical properties is within the knowledge of one of ordinary skill in the art, such as determining cytotoxicity or inhibition of a given target or channel in a cell to determine potential toxicity.

[0113] As used herein, the terms "individual" or "patient," used interchangeably, refer to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, most preferably a human.

[0114] As used herein, the phrase "therapeutically effective amount" refers to that amount of active salt, crystalline form, or pharmaceutical agent that elicits the biological or medicinal response sought in a tissue, system, animal, individual, or human by a researcher, veterinarian, physician, or other clinician.

[0115] As used herein, the term "treating" or "treatment" can refer to one or more of: (1) inhibiting a disease, e.g., inhibiting a disease, condition, or disorder (i.e., halting further progression of the symptoms and / or symptomology) in an individual experiencing or exhibiting the symptoms or symptomology of the disease, condition, or disorder; and (2) ameliorating a disease, e.g., improving a disease, condition, or disorder (i.e., reversing the symptoms and / or symptomology) in an individual experiencing or exhibiting the symptoms or symptomology of the disease, condition, or disorder, such as reducing the severity of the disease.

[0116] In some embodiments, the salts and crystalline forms of the present invention may be useful in preventing the onset of, or reducing the risk of developing, any of the diseases mentioned herein, for example, in preventing the onset of, or reducing the risk of developing, a disease, condition, or disorder in individuals who may be predisposed to the disease, condition, or disorder but who have not yet experienced or exhibited the symptoms or symptomology of the disease.

[0117] Combination therapy I. Immune checkpoint therapy In some embodiments, the PI3K gamma inhibitors provided herein can be used in combination with one or more immune checkpoint inhibitors for the treatment of cancers as described herein. In one embodiment, the combination with one or more immune checkpoint inhibitors as described herein can be used for the treatment of melanoma. The salts and crystalline forms of the present disclosure can be used in combination with one or more immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors of immune checkpoint molecules such as CD20, CD28, CD40, CD122, CD96, CD73, CD47, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, 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, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from 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 disclosure provided herein may be used in combination with one or more agents selected from a KIR inhibitor, a TIGIT inhibitor, a LAIR1 inhibitor, a CD160 inhibitor, a 2B4 inhibitor, and a TGFRbeta inhibitor.

[0118] In some embodiments, the PI3Kγ inhibitors provided herein may 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).

[0119] In some embodiments, the inhibitor of an immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0120] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-1, e.g., an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), durvalumab (Imfinzi®), 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-PD-1 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 anti-cancer agent(s) include antibody therapeutics such as 4-1BB (e.g., urelumab, utomilumab).

[0121] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-L1, 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.

[0122] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-1 and PD-L1, e.g., an anti-PD-1 / PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1 / PD-L1 is MCLA-136.

[0123] In some embodiments, the inhibitor is MCLA-145.

[0124] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884, or CP-675,206.

[0125] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of LAG3, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, or INCAGN2385.

[0126] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TIM3, such as an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

[0127] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of GITR, 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.

[0128] In some embodiments, the inhibitor of an immune checkpoint molecule 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.

[0129] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD20, e.g., an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

[0130] The salts and crystalline forms of the present disclosure may 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 a TGFβ receptor.

[0131] In some embodiments, the PI3K gamma 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.

[0132] As provided throughout, the additional compounds, inhibitors, drugs, etc. may be combined with the present compounds in a single or sequential dosage form, or they may be administered simultaneously or sequentially as separate dosage forms.

[0133] II. Cancer Treatment The growth and survival of cancer cells can be affected by multiple signal transduction pathways. Therefore, combining different enzyme / protein / receptor inhibitors that exhibit different selectivity in the targets that regulate their activity is useful for treating such conditions. Targeting two or more signal transduction pathways (or two or more biological molecules involved in a given signal transduction pathway) can reduce the likelihood of drug resistance occurring in cell populations and / or reduce the toxicity of treatment.

[0134] The salts and crystalline forms of the present disclosure can be used in combination with one or more other enzyme / protein / receptor inhibitors or one or more therapies for the treatment of diseases such as cancer. Examples of diseases and indications treatable with combination therapy include those described herein. Examples of cancer include solid tumors and liquid tumors such as blood cancers.

[0135] For example, one or more additional pharmaceutical agents, such as chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressants, cancer immunotherapeutic agents, metabolic enzyme inhibitors, chemokine receptor inhibitors, and phosphatase inhibitors, as well as 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 WO2006 / 056399. Other agents, such as therapeutic antibodies, may be used in combination with the salts and crystalline forms of the present disclosure for the treatment of PI3K-related diseases, disorders, or conditions. The one or more additional pharmaceutical agents can be administered to a patient simultaneously or sequentially.

[0136] For example, the salts and crystalline forms as disclosed herein can be combined with one or more inhibitors of the following kinases: Akt1, Akt2, Akt3, TGF-βR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IGF-1R, IR-R, PDGFαR for the treatment of cancer and other diseases or disorders described herein. , 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 / Flt2, 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 present disclosure for the treatment of cancer and other diseases and disorders described herein include FGFR inhibitors (FGFR1, FGFR2, FGFR3, or FGFR4, e.g., INCB54828, INCB62079, and INCB63904), JAK inhibitors (JAK1 and / or JAK2, e.g., ruxolitinib, baricitinib, or INCB39110), IDO inhibitors (e.g., epacadostat, NLG919, or BMS-986205), LSD1 inhibitors (e.g., INCB59872 and INCB63904), and cytotoxic T cell proliferation inhibitors (CTL1, CTL2, CTL3, CTL4, CTL54828, CTL62079, and CTL63904). 0003), TDO inhibitors, PI3K-delta inhibitors (e.g., INCB50797 and INCB50465), Pim inhibitors, CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl, and Mer), histone deacetylase inhibitors (HDACs) such as HDAC8 inhibitors, angiogenesis inhibitors, interleukin receptor inhibitors, bromo- and extra-terminal family member inhibitors (e.g., bromodomain inhibitors or BET inhibitors such as INCB54329 and INCB57643), and adenosine receptor antagonists, or combinations thereof.

[0137] 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 is selective for JAK1 over JAK2.

[0138] 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 against c-MET.

[0139] One or more of the following agents may be used in combination with the salts and crystalline forms of the present disclosure, which are presented as a non-limiting list: cytostatics, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epothilone, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, SCH66336, R115777, L778,123, BMS214662, IRESSA™ (gefitinib), TARCEVA™ (erlotinib), antibodies against EGFR, intron, ara-C, adriamycin, cytoxan, gemcitabine, uracil masterbatch. amide, chlormethine, ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, oxaliplatin, leucovirin, ELOXATIN (oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17. alpha-.Ethinyl estradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megstrol acetate, 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, Avastin, HERCEPTIN™ (trastuzumab) Mab), BEXXAR™ (tositumomab), VELCADE™ (bortezomib), ZEVALIN™ (ibritumomab tiuxetan), TRISENOX™ (arsenic trioxide), XELODA™ (capecitabine), vinorelbine, porfimer, ERBITUX™ (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, relozoo fluprexa, fulvestrant, exemestane, ifosfomide, rituximab, C225 (cetuximab), Campath (alemtuzumab), clofarabine, cladribine, afidicolone, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triapine, didox, trimidox, amidox, 3-AP, and MDL-101,731.

[0140] The salts and crystalline forms of the present disclosure can further be used in combination with other methods of treating cancer, such as chemotherapy, radiation therapy, tumor-targeted therapy, adjuvant therapy, immunotherapy, or surgery. Examples of immunotherapies include cytokine treatment (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, adoptive T cell transfer, Toll receptor agonists, STING agonists, oncolytic virotherapy, and immunomodulatory small molecules, including thalidomide or JAK1 / 2 inhibitors. The salts and crystalline forms can be administered in combination with one or more anti-cancer agents, such as chemotherapeutic agents. Examples of chemotherapy drugs include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, bortezomib, intravenous busulfan, oral busulfan, calcitonin, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib ... Lotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, indiragi Terferon alfa 2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab,The present invention includes any of olaparivoxaliplatin, 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.

[0141] Additional examples of chemotherapeutic agents include proteosome inhibitors (eg, bortezomib), thalidomide, revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

[0142] Exemplary steroids include corticosteroids such as dexamethasone or prednisone.

[0143] Exemplary Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC™), nilotinib, dasatinib, bosutinib, and ponatinib, and pharmaceutically acceptable salts. Other exemplary suitable Bcr-Abl inhibitors include compounds of the genera and species disclosed in U.S. Pat. No. 5,521,184, WO 04 / 005281, and U.S. Ser. No. 60 / 578,491, and pharmaceutically acceptable salts thereof.

[0144] Exemplary 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 exemplary suitable Flt-3 inhibitors include compounds such as those disclosed in WO03 / 037347, WO03 / 099771, and WO04 / 046120, and pharmaceutically acceptable salts thereof.

[0145] Exemplary suitable RAF inhibitors include dabrafenib, sorafenib, and vemurafenib, and pharmaceutically acceptable salts thereof. Other exemplary suitable RAF inhibitors include compounds such as those disclosed in WO00 / 09495 and WO05 / 028444, and pharmaceutically acceptable salts thereof.

[0146] Exemplary suitable FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520, and GSK2256098, and pharmaceutically acceptable salts thereof. Other exemplary suitable FAK inhibitors include those compounds disclosed in WO04 / 080980, WO04 / 056786, WO03 / 024967, WO01 / 064655, WO00 / 053595, and WO01 / 014402, and pharmaceutically acceptable salts thereof.

[0147] In some embodiments, the salts and crystalline forms of the present disclosure can be used in combination with one or more other kinase inhibitors, including imatinib, particularly to treat patients who are resistant to imatinib or other kinase inhibitors.

[0148] In some embodiments, the salts and crystalline forms of the present disclosure can be used in combination with chemotherapeutic agents in the treatment of cancer, and can improve therapeutic responses compared to the response to the chemotherapeutic agent alone without exacerbating its toxic effects. In some embodiments, the salts and crystalline forms of the present disclosure can be used in combination with the chemotherapeutic agents provided herein. For example, additional pharmaceutical agents used in the treatment of multiple myeloma can include, but are not limited to, melphalan, melphalan plus prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). Further additional agents used in 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 immunomodulatory agent. 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 immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM). Additive or synergistic effects are desirable outcomes of combining a PI3K inhibitor of the present disclosure with an additional agent.

[0149] In some embodiments, the salts and crystalline forms of the present disclosure can be used in combination with inhibitors of JAK or PI3Kδ.

[0150] The agents may be combined with the compound in a single or continuous dosage form, or the agents may be administered simultaneously or sequentially as separate dosage forms.

[0151] The salts and crystalline forms of the present disclosure can be used in combination with one or more other inhibitors or one or more therapies for the treatment of infectious diseases, including viral, bacterial, fungal, or parasitic infections.

[0152] In some embodiments, a corticosteroid such as dexamethasone is administered to a patient in combination with a compound of the present disclosure, wherein the dexamethasone is administered intermittently rather than continuously.

[0153] The salts and crystalline forms of the present disclosure described herein can be combined with another immunogen, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides, and carbohydrate molecules), cells, and cells transfected with genes encoding immune-stimulating cytokines. Non-limiting examples of tumor vaccines that can be used include peptides of melanoma antigens, such as peptides of gp100, MAGE antigens, Trp-2, MARTI, and / or tyrosinase, or tumor cells transfected to express the cytokine GM-CSF.

[0154] The salts and crystalline forms of the present disclosure described herein can be used in combination with vaccination protocols for the treatment of cancer. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins from viruses implicated in 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 present disclosure can be used in combination with tumor-specific antigens, such as heat shock proteins isolated from the tumor tissue itself. In some embodiments, the salts and crystalline forms can be combined with dendritic cell immunization to activate potent anti-tumor responses.

[0155] The salts and crystalline forms of the present disclosure can be used in combination with bispecific macrocyclic peptides that target Fe alpha or Fe gamma receptor-expressing effector cells to tumor cells. The salts and crystalline forms of the present disclosure can also be used in combination with macrocyclic peptides that activate host immune responsiveness.

[0156] In some further embodiments, combinations of the salts and crystalline forms of the present disclosure with other therapeutic agents can be administered to patients before, during, and / or after bone marrow or stem cell transplantation. The salts and crystalline forms of the present disclosure can be used in combination with bone marrow transplantation for the treatment of various tumors of hematopoietic origin.

[0157] The salts and crystalline forms can be used in combination with vaccines to stimulate immune responses against pathogens, toxins, and self-antigens. Examples of pathogens for which this therapeutic approach may be particularly useful include those for which there are currently no effective vaccines or for which conventional vaccines are not fully effective. These include, but are not limited to, HIV, hepatitis (types A, B, and C), influenza, herpes, giardia, malaria, leishmaniasis, Staphylococcus aureus, and Pseudomonas aeruginosa.

[0158] Viruses causing infectious diseases treatable by the methods of the present disclosure include, but are not limited to, human papillomavirus, influenza, hepatitis A, B, C, or D virus, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory syndrome virus, Ebola virus, measles virus, herpesvirus (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein-Barr virus), flavivirus, echovirus, rhinovirus, coxsackievirus, cornovirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, infectious disease virus, poliovirus, rabies virus, JC virus, and arboviral encephalitis virus.

[0159] Pathogenic bacteria causing infections treatable by the methods of the present disclosure 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 burgdorferi.

[0160] Pathogenic fungi causing infections treatable by the methods of the present disclosure include, but are not limited to, Candida (e.g., albicans, krusei, glabrata, tropicalis), Cryptococcus neoformans, Aspergillus (e.g., fumigatus, niger), Genus Mucorales (e.g., mucor, absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, and Histoplasma capsulatum. Examples of pathogenic parasites that cause infections treatable by the methods of the present disclosure 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.

[0161] Methods for safely and effectively administering most of these chemotherapeutic agents are known to those skilled in the art, and their administration is described in standard literature. For example, the administration of many chemotherapeutic agents is described in the "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.

[0162] Pharmaceutical Preparations and Dosage Forms When used as a pharmaceutical formulation, the compounds of the present disclosure (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 arts and can be administered by various routes, depending on whether local or systemic treatment is desired and the area to be treated. Administration can be topical (including transdermal, epidermal, ophthalmic, and mucosal delivery, 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, or intramuscular injection or infusion, or intracranial, e.g., intrathecal or intraventricular, administration. Parenteral administration can be in the form of a single bolus dose or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

[0163] The present disclosure also includes pharmaceutical compositions containing, as an active ingredient, a compound of the present disclosure or a pharmaceutically acceptable salt thereof in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the compositions are suitable for topical administration. In preparing the compositions of the present disclosure, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed within such a carrier, for example, in the form of a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or liquid vehicles), ointments containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0164] When preparing formulation, before being combined with other components, active compound can be pulverized to obtain suitable particle size.If active compound is substantially insoluble, it can be pulverized to particle size less than 200 mesh.If active compound is substantially water-soluble, particle size can be adjusted by pulverizing to obtain substantially uniform distribution in formulation, for example, about 40 mesh.

[0165] The compounds of the present disclosure can be milled using known milling procedures, such as wet milling, to obtain particle sizes suitable for tableting and other types of formulations.Finely divided (nanoparticulate) preparations of the compounds of the present disclosure can be prepared by processes known in the art.See, for example, International Application No. WO2002 / 000196.

[0166] Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose.The formulation may additionally contain lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl benzoate and propyl hydroxybenzoate; sweeteners; and flavoring agents.The compositions of the present disclosure can be formulated to provide quick, sustained, or delayed release of active ingredients after administration to patients by adopting procedures known in the art.

[0167] The compositions can be formulated in unit dosage form, each dosage containing from about 5 to about 1000 mg (1 g), more usually about 100 to about 500 mg, of the active ingredient. The term "unit dosage form" refers to a physically discrete unit suitable as a unitary dosage for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce a desired therapeutic effect in association with a suitable pharmaceutical excipient.

[0168] In some embodiments, compositions of the present disclosure contain about 5 to about 50 mg of the active ingredient. One of skill in the art will appreciate that this embodies compositions containing about 5 to about 10, about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50 mg of the active ingredient.

[0169] In some embodiments, compositions of the present disclosure contain about 50 to about 500 mg of the active ingredient. One of skill in the art will appreciate that this embodies compositions containing about 50 to about 100, about 100 to about 150, about 150 to about 200, about 200 to about 250, about 250 to about 300, about 350 to about 400, or about 450 to about 500 mg of the active ingredient.

[0170] In some embodiments, compositions of the present disclosure contain about 500 to about 1000 mg of the active ingredient. One of skill in the art will appreciate that this embodies compositions containing about 500 to about 550, about 550 to about 600, about 600 to about 650, about 650 to about 700, about 700 to about 750, about 750 to about 800, about 800 to about 850, about 850 to about 900, about 900 to about 950, or about 950 to about 1000 mg of the active ingredient.

[0171] Similar dosages of the compounds described herein may be used in the methods and uses of the present disclosure.

[0172] The active compounds can be effective over a wide dosage range and are generally administered in a pharmaceutically effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by the physician depending on the relevant circumstances, including the condition to be treated, the route of administration selected, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, etc.

[0173] To prepare solid compositions such as tablets, the principal active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compounds of the present disclosure. When these preformulation compositions are referred to as homogeneous, the active ingredient is typically dispersed evenly 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 unit dosage forms of the type described above, containing, for example, about 0.1 to about 1000 mg of the active ingredient of the present disclosure.

[0174] The tablets or pills of the present disclosure can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which resists disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.

[0175] Liquid forms into which the compounds and compositions of the present disclosure may be incorporated for oral administration or by injection 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.

[0176] 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, compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device, or the nebulizing device may be attached to a face mask, obturator, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered orally or nasally from devices that deliver the formulation in an appropriate manner.

[0177] Topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc. Cream carrier compositions may be based on water combined with glycerol and one or more other components, such as glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water, suitably combined with other components, such as glycerol, hydroxyethylcellulose, etc. In some embodiments, topical formulations contain at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2, or at least about 5% by weight of a compound of the present disclosure. Topical formulations may be suitably packaged, for example, in 100g tubes, optionally with instructions for the treatment of a selected indication, such as psoriasis or other skin conditions.

[0178] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of the administration, such as prophylaxis or treatment, the condition of the patient, the method of administration, etc. In therapeutic applications, compositions may be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the condition of the disease being treated, as well as the judgment of the attending clinician, based on factors such as the severity of the disease, the age, weight, and general health of the patient, etc.

[0179] The compositions administered to patients may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or sterile filtered. Aqueous solutions may be packaged or lyophilized for immediate use, although lyophilized preparations are combined with a sterile aqueous carrier prior to administration. The pH of compound preparations is typically 3-11, more preferably 5-9, and most preferably 7-8. It is understood that certain use of the aforementioned excipients, carriers, or stabilizers may result in the formation of pharmaceutical salts.

[0180] Therapeutic dosages of compounds of the present disclosure can vary depending, for example, on the particular application for which the treatment is given, the manner in which the compound is administered, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound of the present disclosure in a pharmaceutical composition can vary depending on several factors, including dosage, chemical properties (e.g., hydrophobicity), and the route of administration. For example, compounds of the present disclosure can be provided in an aqueous physiological buffer solution containing about 0.1 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 of body weight per day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg of body weight per day. The dosage can depend on variables such as the type and progression of the disease or disorder, the overall health of the particular patient, the relative biological availability of the selected compound, the excipient formulation, and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0181] The compositions of the present disclosure may further comprise one or more additional pharmaceutical agents, such as chemotherapeutic agents, steroids, anti-inflammatory compounds, or immunosuppressants, examples of which are listed herein.

[0182] Labeled Compounds and Assay Methods Another aspect of the present disclosure relates to labeled compounds (e.g., salts and crystalline forms) of the present disclosure that may be useful in both in vitro and in vivo assays, as well as imaging techniques, for localizing and quantifying PI3K in tissue samples, including humans, and for identifying PI3K ligands through inhibitory binding of the labeled compounds (e.g., radiolabeled, fluorescently labeled, etc.). Substitution of one or more atoms of the disclosed compounds may also be useful in generating differentiated ADME (adsorption, distribution, metabolism, and excretion). Accordingly, the present disclosure includes PI3K assays containing such labeled or substituted compounds.

[0183] The present disclosure further includes isotopically labeled compounds of the present disclosure. An "isotopically" or "radiolabeled" compound is a compound of the present disclosure in which one or more atoms have been replaced or substituted with an atom having an atomic mass or mass number different from that typically found in nature (i.e., naturally occurring). Suitable radionuclides that can be incorporated into compounds of the present disclosure include: 2 H (also written as D for deuterium), 3 H (also written as T for tritium), 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 For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced by a deuterium atom (e.g., C of formula (I) 1~6 One or more hydrogen atoms of an alkyl group can be optionally replaced with a deuterium atom, such as replacing -CH with -CD.) In some embodiments, alkyl groups of the formulas and / or forms disclosed herein can be all deuterated.

[0184] One or more constituent atoms of the compounds provided herein may be replaced or substituted with an isotope of the atom at natural or non-natural abundance. In some embodiments, the compounds contain at least one deuterium atom. For example, one or more hydrogen atoms in the compounds provided herein may be replaced or substituted with a deuterium atom (e.g., C 1-6One or more hydrogen atoms of an alkyl group can be replaced with a deuterium atom, such as replacing -CH with -CD. In some embodiments, the compound contains two or more deuterium atoms. In some embodiments, the compound contains 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.

[0185] In some embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms bonded to carbon atoms of any Formula A substituent are each optionally replaced by a deuterium atom.

[0186] Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971); The Renaissance of H / D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds can be used in a variety of studies, such as NMR spectroscopy, metabolic studies, and / or assays.

[0187] Substitution with heavier isotopes, such as deuterium, can provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferable in some situations. (See, e.g., 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 can provide one or more therapeutic advantages.

[0188] The radionuclide incorporated into the radiolabeled compound will depend on the particular application of the radiolabeled compound. For example, for in vitro PI3K labeling and competition assays: 3 H, 14 C. 82 Br, 125 I, 131 I, or 35 Compounds incorporating S may be useful. For radio-imaging applications, 11 C. 18 F, 125 I, 123 I, 124 I, 131 I, 75 Br, 76 Br, or 77 Br may be useful.

[0189] A "radiolabel" or "labeled compound" is understood to be a compound that incorporates at least one radionuclide. In some embodiments, the radionuclide is 3 H, 14 C. 125 I, 35 S, and 82 Br.

[0190] The present disclosure can further include synthetic methods for incorporating radioisotopes into the compounds of the present disclosure. Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and one of ordinary skill in the art would readily recognize methods applicable to the compounds of the present disclosure.

[0191] The labeled compounds of the present disclosure can be used in screening assays to identify / evaluate compounds. For example, a newly synthesized or identified labeled compound (i.e., a test compound) can be evaluated for its ability to bind to PI3K by monitoring its concentration change when contacted with PI3K through tracking the label. For example, a test compound (labeled) can be evaluated for its ability to reduce the binding of another compound (i.e., a standard compound) known to bind to PI3K. Thus, the ability of a test compound to compete with a standard compound for binding to PI3K is directly correlated with its binding affinity. Conversely, in some other screening assays, the standard compound is labeled, and the test compound is unlabeled. Therefore, to evaluate the competition between the standard compound and the test compound, the concentration of the labeled standard compound is monitored, thereby determining the relative binding affinity of the test compound.

[0192] kit The present disclosure also includes pharmaceutical kits useful in the treatment or prevention of PI3K-related diseases or disorders, such as cancer, which include one or more containers housing pharmaceutical compositions comprising therapeutically effective amounts of the salts and crystalline forms of the present disclosure. As will be apparent to those of skill in the art, such kits can optionally further include one or more of a variety of conventional pharmaceutical kit components, such as, for example, a container with one or more pharmaceutically acceptable carriers, additional containers, etc. Instructions, either as an insert or label, indicating the amounts of components to be administered, administration guidelines, and / or guidelines for mixing the components, can also be included in the kit.

[0193] The present invention will be described in more detail using specific examples.The following examples are provided for illustrative purposes and are not intended to limit the present invention in any way.Those skilled in the art will easily recognize various non-critical parameters that can be changed or modified to achieve essentially the same results.The salts and crystalline forms of the examples have been found to be PI3Kγ inhibitors by at least one assay described herein. [Example]

[0194] Preparative LC-MS purification of some of the prepared compounds was carried out on a Waters mass spectrometer. The basic equipment settings, protocols, and control software for the operation of 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 typically subjected to analytical liquid chromatography mass spectrometry (LCMS) for purity analysis under the following conditions: Apparatus: Agilent 1100 series LC / MSD; Column: Waters Sunfire™ C 18 5 μm, 2.1×50 mm, buffer: Mobile phase A: 0.025% TFA in water and mobile phase B: acetonitrile; gradient from 2% to 80% B in 3 minutes at a flow rate of 2.0 mL / min.

[0195] Some of the prepared compounds were also separated on a preparative scale by reversed-phase high-performance liquid chromatography (RP-HPLC) with MS detection or flash chromatography (silica gel), as shown in the examples. Typical preparative reversed-phase high-performance liquid chromatography (RP-HPLC) column conditions are as follows: Purification at pH=2: Waters Sunfire™ C 18 , 5 μm, 30 × 100 mm or Waters XBridge™ C 18 , 5 μm, 30 × 100 mm column, elution with mobile phase A: 0.1% TFA (trifluoroacetic acid) in water and mobile phase B: acetonitrile; flow rate was 60 mL / min, and the separation gradient was optimized for each compound using a Compound Specific Method Optimization protocol as 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)).

[0196] pH=10 purification: Waters XBridge (trademark) C 18 A 5 μm, 30 × 100 mm column was used, eluting with mobile phase A: 0.1% NH OH in water and mobile phase B: acetonitrile; the flow rate was 60 mL / min, and the separation gradient was optimized for each compound using a Compound Specific Method Optimization protocol as described in the literature (see, e.g., “Preparative LCMS Purification: Improved Compound Specific Method Optimization,” K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)).

[0197] In the following examples, X-ray powder diffraction analyses were performed on a Bruker D8 Advance EC X-ray powder diffractometer (XRPD) instrument using the following parameters: a 1.5418 Å Cu source and a LYNXEYE™ detector and an X-ray output of 40 KV, 25 mA. Sample powder was dispersed in a zero-background sample holder. Typical measurement conditions were: start angle -3°; end angle -30°; sample collection -0.015°; and scan rate -2° / min.

[0198] Differential scanning calorimetry (DSC) was performed on a TA Instrument differential scanning calorimeter, Discovery DSC2500, equipped with an autosampler. Typical experimental conditions were: 20–300 °C at 10 °C / min, nitrogen gas flow of 50 mL / min, using aluminum sample pans.

[0199] Thermogravimetric analysis (TGA) was performed on a TA Instrument Thermogravimetric Analyzer TGA5500 equipped with an autosampler under the following conditions: 10°C / min ramp from 25°C to 600°C; nitrogen gas at 25 mL / min balance purge flow; and platinum sample pan.

[0200] Example 1. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide [ka] Step 1. 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-ol [ka] 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 at 0 °C for several minutes. 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 at ambient temperature for 30 minutes. 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 tan 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. C9H7BrF3(M-OH) + LCMS for: m / z = 251.0, 253.0; Found: 250.9, 252.8.

[0201] Step 2. 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-one [ka] A mixture of 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-ol (8.41 g, 31.3 mmol) in dichloromethane (125 mL) at 0° C. was treated with Dess-Martin periodinane (19.9 g, 46.9 mmol) and stirred at RT for 2.5 h. The reaction mixture was concentrated (by rotary evaporation with a water bath set at 30° C.) to an oily solid, which was diluted with diethyl ether (200 mL), causing more solid to precipitate. The mixture was filtered through Celite®, and the Celite® was rinsed with additional diethyl ether (200 mL). The filtrate was washed with saturated sodium bicarbonate solution (3×200 mL) and brine, dried over sodium sulfate, filtered, and concentrated to give an oily solid. The oily solid was partitioned between diethyl ether (150 mL) and water (100 mL). The organic layer was separated, 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. C9H7BrF3O (M+H) + LCMS for: m / z = 267.0, 269.0; Found: 267.1, 268.9.

[0202] Step 3. 2-(3-bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanenitrile [ka] 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 h. Due to an exotherm following the addition of 18-crown-6, the reaction was cooled in an ice bath. The reaction mixture was concentrated (by rotary evaporation with a water bath set at 28 °C) to give a reddish-brown 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 h. 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 (8.70 g, 99.8%) as an orange solid, which was used without further purification. C9H7BrF3O (M-CN) + LCMS for: m / z = 267.0, 269.0; Found: 266.9, 269.0.

[0203] Step 4. 2-(3-bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (second-eluting enantiomer) [ka] A solution of 2-(3-bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanenitrile (8.70 g, 29.6 mmol) in 1,4-dioxane (59.2 mL) at 0° C. was treated with concentrated HCl (9.00 mL, 108 mmol) pre-cooled in an ice bath. HCl gas was bubbled through the reaction mixture while stirring at 0° C. for 45 minutes. The cooling bath was removed, and the reaction mixture was stirred at room temperature for 61 hours. Nitrogen was bubbled through the reaction mixture for 10 minutes to remove some of the HCl, then the reaction mixture was 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 any 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. Purification by flash column chromatography using MTBE (0% to 60%) in hexanes afforded the racemic product as a yellow, oily solid. The racemic mixture was separated by preparative chiral HPLC (Phenomenex Lux Amylose-1 [21.2 × 250 mm, 5 micrometers], eluting with 95% ethanol in hexanes at a flow rate of 18 mL / min, loading approximately 100 mg in 2 mL of ethanol) to afford the desired second-eluting enantiomer (4.50 g, 48.8%) as a viscous, yellow oil. The first enantiomer to elute had a retention time of 4.0 min. The second enantiomer to elute had a retention time of 5.3 min.

[0204] Second eluting enantiomer: 1 H NMR (400 MHz, DMSO-d6) δ 7.85 (d, J = 1.9 Hz, 1H), 7.75 (s, 1H), 7.67 (s, 1H), 7.63 - 7.53 (m, 2H), 7.41 (d, J = 8.1 Hz, 1H), 2.35 (s, 3H). C 10 H 10 BrF3NO2(M+H) + LCMS for: m / z = 312.0, 314.0; Found: 312.0, 314.0.

[0205] 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 [ka] 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 enantiomer) 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), filtered through Celite®, and rinsed with additional ethyl acetate (100 mL). The filtrate was washed with brine, dried over sodium sulfate, filtered, and concentrated to a brown foam. Purification by flash column chromatography using MTBE in hexanes (0% to 100%) gave the desired product (3.35 g, 81.5%) as a viscous yellow foam. 1 H NMR (400 MHz, DMSO-d6) δ 7.96 (d, J = 2.2 Hz, 1H), 7.63 (dd, J = 7.9, 2.1 Hz, 1H), 7.58 (s, 1H), 7.54 (s, 1H), 7.51 - 7.40 (m, 1H), 7.21 (d, J = 8.2 Hz, 1H), 2.46 (s, 3H), 1.30 (s, 12H). C 16 H 22 BF3NO4(M+H) + LCMS for: m / z = 360.2; Found: 360.1.

[0206] Step 6. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide 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 potassium carbonate in water (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 an additional 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 the Celite was 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 to a brown foam. Purification by flash column chromatography using MeOH (0% to 10%) in dichloromethane gave the desired product as a red / brown foam, which was not completely pure. This material was repurified by flash column chromatography using MeOH (0% to 15%) in dichloromethane to give the desired product as an orange / brown foam, which was still not completely pure. This material was repurified by flash column chromatography using ethyl acetate (with 5% MeOH) in hexanes (0% to 100%) to give the desired product as a white foam, which was still impure. This material was repurified by flash column chromatography using acetonitrile (containing 5% MeOH) in dichloromethane (0% to 100%) to give the desired product (4.67 g, 40.4%) as a white foam. 1H NMR (600 MHz, DMSO-d6) δ 7.79 (s, 1H), 7.76 - 7.71 (m, 2H), 7.71 - 7.64 (m, 4H), 7.61 (d, J = 3.5 Hz, 2H), 7.51 (d, J = 8.2 Hz, 1H), 2.23 (s, 3H). C 17 H 14 F6N5O2(M+H) + LCMS for: m / z = 434.1; Found: 434.1.

[0207] 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) 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) and heptane (1.34 mL) was added dropwise. After 2 mL was added, the solid did not completely dissolve, and some remained at the bottom of the vial. After nearly all of the solid dissolved, new solid formed on the wall of the vial. After stirring at 80° C. for 2 hours, more solid formed. After cooling to ambient temperature, the solid was filtered and washed with heptane. The solid was collected and dried under vacuum for 30 minutes 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.

[0208] Form IA was confirmed as a crystalline solid by XRPD analysis. The XRPD pattern of Form IA is shown in Figure 1, and the peak data is provided in Table 1 below. [Table 1]

[0209] DSC analysis of Form IA revealed a single endothermic peak with an onset temperature of 191.9° C. and a maximum of 193.2° C. The DSC thermogram is shown in FIG.

[0210] TGA analysis of Form IA revealed significant weight loss above 200° C. due to decomposition of the sample. The TGA thermogram is shown in FIG.

[0211] Form IA was identified as an anhydrous, unsolvated crystalline form.

[0212] 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) 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 cooled to ambient temperature and stirred for 1 hour. The solid was collected by filtration and air-dried to provide 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (Form IIA).

[0213] Form IIA was confirmed as a crystalline solid by XRPD analysis. The XRPD pattern of Form IIA is shown in Figure 4, and the peak data is shown in Table 2 below. [Table 2]

[0214] DSC analysis of Form IIA revealed a single endothermic peak with an onset temperature of 177.2° C. and a maximum of 179.7° C. The DSC thermogram is shown in FIG.

[0215] TGA analysis of Form IIA revealed significant weight loss above 200° C. due to decomposition of the sample. The TGA thermogram is shown in FIG.

[0216] Form IIA was identified as an anhydrous, non-solvated crystalline form.

[0217] 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) 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, a MeOH solvate, was dried under vacuum at 60° C. overnight to provide 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (Form IIIA).

[0218] Form IIIA was confirmed as a crystalline solid by XRPD analysis. The XRPD pattern of Form IIIA is shown in Figure 7, and the peak data is shown in Table 3 below. [Table 3]

[0219] DSC analysis of Form IIIA revealed a single endothermic peak with an onset temperature of 134.3° C. and a maximum of 143.0° C. The DSC thermogram is shown in FIG.

[0220] TGA analysis of Form IIIA revealed significant weight loss above 200° C. due to decomposition of the sample. The TGA thermogram is shown in FIG.

[0221] Form IIIA was identified as an anhydrous, non-solvated crystalline form.

[0222] Examples 5-6. 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 (Enantiomers 1 and 2) [ka] Step 1. 6,8-Dibromo-3-iodoimidazo[1,2-a]pyrazine [ka] 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 h. The reaction mixture was concentrated in vacuo. The resulting solid was taken up in dichloromethane (DCM). The organic layer was washed successively with water and saturated NaSO (aq). The organic layer was then dried over NaSO, filtered, and concentrated to give the title compound as a pale yellow solid (0.64 g, 88%). C6H3Br2IN3 (M+H) + LCMS for: calculated m / z = 401.8, 403.8, 405.8; found 401.8, 403.7, 405.6.

[0223] Step 2. 6-Bromo-3-iodo-N-(4-methoxybenzyl)imidazo[1,2-a]pyrazin-8-amine [ka] 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)methanamine (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. 14 H 13 BrIN4O (M+H) + LCMS for: calculated m / z = 458.9, 460.9; found 459.0, 461.0.

[0224] Step 3. 6-Bromo-3-iodoimidazo[1,2-a]pyrazin-8-amine trifluoroacetate [ka] A solution of 6-bromo-3-iodo-N-(4-methoxybenzyl)imidazo[1,2-a]pyrazin-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, diluted with acetonitrile (2×), and then reconcentrated. The solid was diluted with ethyl acetate (12 mL) and stirred at room temperature for 1 hour. The slurry was diluted with hexane (12 mL) dropwise 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. C6H5BrIN4 (M+H) + LCMS for: calculated m / z = 338.9, 340.9; found 338.8, 340.8.

[0225] Step 4. 2-(3-bromo-4-methylphenyl)-1,1,1-trifluoropropan-2-ol [ka] A solution of 1-(3-bromo-4-methylphenyl)ethan-1-one (1.20 g, 5.63 mmol) [Aldrich, 579734] in tetrahydrofuran (22.5 mL) at 0 °C was treated with trimethyl(trifluoromethyl)silane (1.00 mL, 6.76 mmol) [Aldrich, 488712] and stirred at 0 °C for 5 minutes. The reaction mixture was treated with 1.0 M tetra-n-butylammonium fluoride in tetrahydrofuran (0.282 mL, 0.282 mmol) at 0 °C and stirred at room temperature for 1 hour. The reaction mixture was cooled to 0 °C and treated with additional 1.0 M tetra-n-butylammonium fluoride in tetrahydrofuran (6.76 mL, 6.76 mmol) and stirred at room temperature for 30 minutes. The reaction mixture was diluted with ethyl acetate (100 mL) and washed with brine (2 × 75 mL). The organic layer was separated, dried over sodium sulfate, filtered, and concentrated to give the crude residue. Purification by flash column chromatography using ethyl acetate in hexanes (0% to 30%) gave the desired product (1.54 g, 96.7%) as a yellow oil. 10 H9BrF3(M-OH) + LCMS for: m / z = 265.0, 267.0; Found: 264.9, 267.0.

[0226] 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 [ka] A mixture of 2-(3-bromo-4-methylphenyl)-1,1,1-trifluoropropan-2-ol (0.252 g, 0.890 mmol), bis(pinacolato)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 an additional 5 minutes, and heated in a microwave at 135° C. for 20 minutes. The reaction mixture was diluted with ethyl acetate and filtered through a 0.5 micrometer cartridge, which was 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 hexanes (0% to 50%) gave the desired product (272 mg, 92.5%) as a colorless oil. 16 H 23 BF3O3(M+H) + LCMS for: m / z = 331.2; Found: 331.2.

[0227] Step 6. 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-1,1,1-trifluoropropan-2-ol [ka] A mixture of 6-bromo-3-iodoimidazo[1,2-a]pyrazin-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 sodium carbonate in water (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 (2x). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated to give the crude residue. Purification by flash column chromatography using methanol in dichloromethane (0% to 2%) afforded the desired product (610 mg, 77.8%) as a white foam. 16 H 15 BrF3N4O (M+H) + LCMS for: m / z = 415.0, 417.0; Found: 415.0, 417.0.

[0228] Step 7. Methyl 8-amino-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylate [ka] 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 minutes. The reaction mixture was treated with Pd(dppf)CHCl (0.049 g, 0.060 mmol), degassed with nitrogen for an additional 5 minutes, saturated with CO by bubbling the gas under the reaction surface 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 a brown oil. Purification by flash column chromatography using methanol (0% to 4%) in dichloromethane gave the desired product (158 mg, 66.5%) as a tan oily solid. 18 H 18 F3N4O3(M+H) + LCMS for: m / z = 395.1; Found: 395.1.

[0229] 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 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 filtered through a pad of Celite®. After rinsing with MeOH (2×), the filtrate was concentrated to a tan oil. Purification by silica gel chromatography (0-5% MeOH / DCM) afforded the title compound as an oily solid (26 mg, 28%), which was a mixture of enantiomers. The racemic mixture was separated by preparative chiral HPLC (Phenomenex Lux Amylose-1 [21.2 × 250 mm, 5 micrometers], eluting with 12% ethanol in hexane at a flow rate of 18 mL / min, loading approximately 8 mg in 800 μL of ethanol). The first peak to elute had a retention time of 11.9 minutes (Example 1; Enantiomer 1). The second peak to elute had a retention time of 16.1 minutes (Example 2, Enantiomer 2).

[0230] Example 5 (Enantiomer 1): 1 H NMR (400 MHz, DMSO-d6) δ 8.08 (t, J = 6.1 Hz, 1H), 7.70 (d, J = 2.8 Hz, 2H), 7.65 (d, J = 8.1 Hz, 1H), 7.58 (s, 1H), 7.49 (d, J = 8.2 Hz, 1H), 7.36 (s, 2H), 6.65 (s, 1H), 4.65 (s, 1H), 3.22 (d, J = 6.1 Hz, 2H), 2.15 (s, 3H), 1.70 (s, 3H), 1.09 (s, 6H). C 21 H 25F3N5O3(M+H) + LCMS for: m / z = 452.2; Found: 452.1.

[0231] Example 6 (Enantiomer 2): 1 H NMR (600 MHz, DMSO-d6) δ 8.08 (t, J = 6.0 Hz, 1H), 7.70 (d, J = 3.0 Hz, 2H), 7.65 (d, J = 8.2 Hz, 1H), 7.58 (s, 1H), 7.49 (d, J = 8.2 Hz, 1H), 7.36 (s, 2H), 6.65 (s, 1H), 4.65 (s, 1H), 3.22 (d, J = 6.1 Hz, 2H), 2.15 (s, 3H), 1.70 (s, 3H), 1.09 (s, 6H). C 21 H 25 F3N5O3(M+H) + LCMS for: m / z = 452.2; Found: 452.2.

[0232] 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) 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 Example 6, Step 8; 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 RT 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%).

[0233] Form IB was confirmed as a crystalline solid by XRPD analysis. The XRPD pattern of Form IB is shown in Figure 10, and the peak data is shown in Table 4 below. [Table 4]

[0234] DSC analysis of Form IB revealed a single endothermic peak with an onset temperature of 172.2° C. and a maximum of 174.2° C. The DSC thermogram is shown in Figure 11. Form IB was confirmed as an anhydrous, unsolvated crystalline form.

[0235] 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) 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 a solid persisted. The mixture was heated at 80° C. for 30 min and stirred at RT 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%).

[0236] Form IIB was confirmed as a crystalline solid by XRPD analysis. The XRPD pattern of Form IIB is shown in Figure 12, and the peak data is shown in Table 5 below. [Table 5]

[0237] DSC analysis of Form IIB revealed a single endothermic peak with an onset temperature of 161.7° C. and a maximum of 165.4° C. The DSC thermogram is shown in Figure 8. Form IIB was confirmed as an anhydrous, nonsolvated crystalline form.

[0238] Example 9. 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 [ka] Step 1. 1-(4-(methyl-d3)phenyl)ethan-1-one [ka] 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, treated with iodomethane-d3 (1.44 mL, 23.2 mmol), and stirred at 45 °C overnight. The reaction mixture was cooled to rt 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 give the desired product (546 mg, 65.3%) as a yellow oil, which was used without further purification. 1 H NMR (400 MHz, CDCl3) δ 7.86 (d, J= 8.0 Hz, 2H), 7.26 (d, J= 8.0 Hz, 2H), 2.58 (s, 3H). C9H8D3O (M+H) + LCMS for: m / z = 138.1; Found: 138.1.

[0239] Step 2. 1-(3-bromo-4-(methyl-d3)phenyl)ethan-1-one [ka] A suspension of aluminum chloride (13.6 g, 102 mmol) in dichloromethane (24 mL) was treated with 1-(4-(methyl-d3)phenyl)ethan-1-one (6.35 g, 46.3 mmol) dropwise via syringe over 5 minutes. The remaining material in the syringe was rinsed with dichloromethane (7.0 mL) and added dropwise to the reaction mixture. After the initial exotherm, the reaction mixture was cooled to rt for 3 minutes, stirred at 35° C. for 5 minutes, and treated with bromine (2.38 mL, 46.3 mmol) dropwise over 5 minutes. The reaction mixture was stirred for 25 minutes before being slowly added to a mixture of dichloromethane (50 mL), 1 N HCl (100 mL), and ice. The remaining reaction mixture was rinsed into a dichloromethane / HCl / ice mixture with additional dichloromethane. The mixture was warmed to room temperature (rt), and the layers were separated. The aqueous layer was extracted with dichloromethane (2 x 75 mL). The combined organic layers were washed with saturated sodium bicarbonate and brine. The sodium bicarbonate and brine washes contained the product and were combined, acidified with 1 M HCl, and extracted with dichloromethane (2 x 50 mL). All organic layers were combined, dried over magnesium sulfate, filtered, and concentrated to a yellow oil. Purification by flash column chromatography using ethyl acetate in hexanes (0% to 15%) afforded the desired product (9.08 g, 90.8%) as a pale yellow solid. 1 H NMR (400 MHz, CDCl3) δ 8.11 (d, J = 1.8 Hz, 1H), 7.79 (dd, J = 7.9, 1.8 Hz, 1H), 7.32 (d, J = 7.9 Hz, 1H), 2.57 (s, 3H). C9H7D3BrO (M+H) + LCMS for: m / z = 216.0, 218.0; Found: 216.0, 218.0.

[0240] Step 3. 2-(3-bromo-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol [ka] A solution of 1-(3-bromo-4-(methyl-d3)phenyl)ethan-1-one (9.08 g, 42.0 mmol) in tetrahydrofuran (168 mL) at 0° C. was treated with trimethyl(trifluoromethyl)silane (8.07 mL, 58.8 mmol) [Aldrich, 488712] and stirred at 0° C. for 5 minutes. The reaction mixture was treated with 1.0 M tetra-n-butylammonium fluoride in tetrahydrofuran (2.10 mL, 2.10 mmol) at 0° C. and stirred at room temperature for 1 hour. The reaction mixture was treated with 1.0 M tetra-n-butylammonium fluoride in tetrahydrofuran (12.6 mL, 12.6 mmol) and water (9.8 mL) and stirred at room temperature for 30 minutes. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3×50 mL). 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 ethyl acetate in hexanes (0% to 20%) afforded the desired product (13.3 g, 111%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.79 - 7.72 (m, 1H), 7.48 - 7.35 (m, 1H), 7.24 (s, 1H), 2.41 (br s, 1H), 1.76 (s, 3H).

[0241] Step 4. 1,1,1-trifluoro-2-(4-(methyl-d3)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ol [ka] 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 2-(3-bromo-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol (13.3 g, 41.8 mmol). The remaining 2-(3-bromo-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol was rinsed with dioxane (106 mL) and added to the reaction mixture, which was 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 an additional 10 minutes, and stirred at 100 °C overnight. The reaction mixture was cooled to rt, 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 filtered through Celite® and rinsed with THF and ethyl acetate. The filtrate was washed with 1:1 water / brine (300 mL). The aqueous layer was re-extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered, and concentrated to a brown oil. Purification by flash column chromatography using MTBE (0% to 20%) in hexanes afforded the desired product (14.4 g, 84.7%) as a pale yellow oil. 1 H NMR (400 MHz,CDCl3) δ 7.93 (d, J = 2.3 Hz, 1H), 7.55 - 7.45 (m, 1H), 7.19 (d, J = 8.1 Hz, 1H), 2.43 (br s, 1H), 1.77 (s, 3H), 1.34 (s, 12H). C 16 H 20 D3BF3O3(M+H) + LCMS for: m / z = 334.2; Found: 334.3.

[0242] Step 5. 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol (racemic mixture) [ka] A solution of 1,1,1-trifluoro-2-(4-(methyl-d3)-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 treated with 6-bromo-3-iodoimidazo[1,2-a]pyrazin-8-amine (12.1 g, 35.6 mmol), 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 an additional 5 minutes. The reaction mixture was treated with 1.0 M potassium carbonate in water (107 mL, 107 mmol), degassed with nitrogen for 5 minutes, and stirred at 80 °C overnight. The reaction mixture was cooled to rt and filtered through Celite®. The Celite® was rinsed with ethyl acetate and water. The filtrate was diluted with water (150 mL) and extracted with ethyl acetate (3×100 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% to 5%) and repurification by flash column chromatography using ethyl acetate in hexanes (0% to 100%) gave the desired product (13.8 g, 92.8%). C 16 H 12 D3BrF3N4O (M+H) + LCMS for: m / z = 418.1, 420.1; Found: 418.0, 420.0.

[0243] Step 6. Second-eluting enantiomer of 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol [ka] The racemic mixture of 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol was separated by preparative chiral HPLC (Phenomenex Lux Amylose-1 [21.2 × 250 mm, 5 micrometers], eluting with 20% ethanol in hexane at a flow rate of 20 mL / min, loading approximately 200 mg in 4 mL of ethanol). The first peak to elute had a retention time of 9.6 minutes. The second peak to elute had a retention time of 14.6 minutes.

[0244] Peak 2: 1 H NMR (400 MHz, DMSO-d6) δ 7.66–7.59 (m, 2H), 7.59–7.53 (m, 3H), 7.46 (d, J = 8.1 Hz, 1H), 7.25 (s, 1H), 6.66 (s, 1H), 1.71 (s, 3H). LCMS for CHDBrFNO (M+H): m / z = 418.1, 420.1; found: 418.0, 420.0.

[0245] Step 7. Methyl 8-amino-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylate (single enantiomer prepared) [ka] 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. The reaction mixture was treated with Pd(dppf)2CHCl2 (0.998 g, 1.22 mmol), degassed with nitrogen for an additional 5 min, saturated with CO by bubbling gas under the reaction surface for 3 min, 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% to 5%) in dichloromethane afforded the desired product (4.49 g, 92.4%) as an orange solid. 18 H 15 D3F3N4O3(M+H) + LCMS for: m / z = 398.1; Found: 398.3.

[0246] Step 8. 8-Amino-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid (single enantiomer prepared) [ka] A solution of methyl 8-amino-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylate (4.49 g, 11.3 mmol) (single enantiomer from Step 7) in methanol (113 mL) was treated with 1.0 M sodium hydroxide (56.5 mL, 56.5 mmol) 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 layers were extracted with additional 1.0 M sodium hydroxide (3 × 20 mL). The combined basic aqueous layers were adjusted to pH ∼5 with citric acid (7.6 g). The aqueous layer was extracted with dichloromethane (2 × 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 tan solid, which was used without further purification. 1 H NMR (400 MHz, DMSO-d6) δ 7.76 (s, 1H), 7.71 (s, 1H), 7.65 (dd, J = 8.2, 2.0 Hz, 1H), 7.59 (d, J = 2.0 Hz, 1H), 7.49 (d, J = 8.1 Hz, 1H), 7.30 (br s, 2H), 6.66 (s, 1H), 1.71 (s, 3H). C 17 H 13 D3F3N4O3(M+H) + LCMS for: m / z = 384.1; Found: 384.2.

[0247] Step 9. 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 A solution of 8-amino-3-(2-(methyl-d3)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid (4.06 g, 10.6 mmol) (single enantiomer from Step 8) in DMF (106 mL) was treated with 1-amino-2-methylpropan-2-ol (1.44 g, 16.2 mmol) [Ark Pharm, AK-37803] and HATU (6.16 g, 16.2 mmol), stirred for 15 min, treated with triethylamine (4.43 mL, 31.8 mmol), and stirred at room temperature for 3.5 h. The reaction mixture was diluted with water (500 mL) and brine (100 mL) and extracted with ethyl acetate (3 × 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 a tan oil. Purification by flash column chromatography using methanol (0% to 5%) in dichloromethane gave the desired product (4.28 g, 89.0%) as a foam. 1 H NMR (600 MHz, DMSO-d6) δ 8.14 - 8.05 (m, 1H), 7.74 - 7.69 (m, 2H), 7.66 (d, J = 7.9 Hz, 1H), 7.62 - 7.54 (m, 1H), 7.50 (dd, J = 8.2, 2.0 Hz, 1H), 7.38 (s, 2H), 6.67 (s, 1H), 4.67 (s, 1H), 3.23 (d, J = 5.6 Hz, 2H), 1.71 (s, 3H), 1.10 (s, 6H). C 21 H 22 LCMS for D3F3N5O3 (M+H)+: m / z = 455.2; Found: 455.2.

[0248] 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) A round-bottom flask was charged with 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 (4.60 g, 10.1 mmol) and isopropyl acetate (25.5 mL) and heated to 80° C. The mixture was stirred at 80° C., and a solid began to form within 5 minutes. The mixture was stirred at 80° C. for 1 hour. Heating was discontinued, and the mixture was stirred for 1 hour while cooling to rt. The mixture was treated with heptane (25.5 mL) dropwise 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 under reduced pressure at 60° C. for 24 hours to give 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 (Form I) (4.16 g, 90.4%).

[0249] Form IC was confirmed as a crystalline solid by XRPD analysis. The XRPD pattern of Form IC is shown in Figure 14, and the peak data is shown in Table 6 below. [Table 6]

[0250] DSC analysis of Form IC revealed a single endothermic peak with an onset temperature of 173.4° C. and a maximum of 179.0° C. The DSC thermogram is shown in Figure 15. Form IC was confirmed as an anhydrous, nonsolvated crystalline form.

[0251] Example 11. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide (HBr) salt 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 6 M aqueous HBr solution (1.2 equivalents) was added to the solution and mixed thoroughly. The solution was evaporated at room temperature to give HBr salt crystals.

[0252] 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 Crystal data: C35 H32 Br2 F12 N10 O5, from methanol, colorless, irregular plate, approximately 0.450 x 0.210 x 0.060 mm, monoclinic, C2, a = 20.055(7) Å, b = 10.115(4) Å, c = 21.363(8) Å, beta = 94.953(7), volume = 4318(3) Å 3 , Z=4, T=-40℃, formula weight=1060.52, density=1.631g / cm 3 , μ(Mo)=1.98mm -1 .

[0253] Data Collection: Data collection was performed using a Bruker SMART APEX-II CCD system, MoK alpha radiation, standard focus 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 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, 2theta range = 4.51 to 55.43°, completeness for 2theta 55.43 = 99.60%, R(int-xl) = 0.0672, SADABS correction applied.

[0254] Analysis and refinement: The crystal structure was analyzed using XS (Shelxtl) and refined using the shelxtl software package. 2 Full matrix least squares method for scattering coefficients in Int.Tab.Vol C Tables 4.2.6.8 and 6.1.1.4, number of data = 9756, number of constraints = 1, number of parameters = 584, data / parameter ratio = 16.71, goodness of fit for F2 = 1.14, R-index [I>4 sigma (I)] R1 = 0.0648, wR2 = 0.1560, R-index (all data) R1 = 0.1004, wR2 = 0.1719, maximum difference between peaks and holes = 1.795 and -0.642 e / Å 3 The refinement was performed with a flack parameter = 0.038 (6). All hydrogen atoms were idealized using the riding model. Table 7 shows the atomic coordinates (×10 4 ) and equivalent isotropic displacement parameter (Å 2 x10 3 ) where U(eq) is defined as one-third of the locus of the orthogonal Uij tensor. Table 8 shows the bond lengths [Å] and angles [deg]. Table 9 shows the anisotropic displacement parameters (Å 2 x10 3 ) is shown.

[0255] Results: 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 bromides for charge balancing, and one methanol solvent molecule, as shown in Figures 16A-16B. Enantiomeric assignment was based on a refined Flack parameter of 0.038 (6). This study determined the absolute configurations of the chiral centers C15=S- and C35=S-. [Table 7-1] [Table 7-2] [Table 8-1] [Table 8-2] [Table 8-3] [Table 8-4] [Table 9-1] [Table 9-2]

[0256] Example A. THP-1 RPS6 ELISA Assay 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 containing 10% FBS (Gibco / Life Technologies, Carlsbad, CA). For the assay, THP-1 cells were serum-starved overnight in RPMI and then incubated in RPMI (2 x 10 cells in 90 μL) in the presence or absence of a range of test compounds. 5Cells (cells / well) are plated into 96-well flat-bottom tissue culture-treated plates (Corning, Corning, NY). Covered plates are incubated at 37°C and 5% CO2 for 2 hours, then treated with or without 10 nM MCP-1 (MYBioSource, San Diego, CA) for 15 minutes at 37°C and 5% CO2. Plates are centrifuged at 1600 RPM, and the supernatant is removed. Cells are lysed for 30 minutes on wet ice in lysis buffer (Cell Signaling, Danvers, MA) containing protease inhibitors (Calbiochem / EMD, Germany), PMSF (Sigma, St Louis MO), and HALTS (Thermo Fisher, Rockford, IL). Cell lysates are frozen at -80°C before testing. Lysates are tested using a human / mouse / rat Phospho-RPS6 ELISA (R&D Systems, Inc. Minn, MN). The plate is read using a microplate reader (SpectraMax M5 - Molecular Devices, LLC Sunnyvale, CA) set at 450 nm with wavelength correction at 540. 50 is determined by curve fitting of percent inhibition of inhibitor against the logarithm of inhibitor concentration using GraphPad Prism 5.0 software.

[0257] Example B. Scintillation Proximity Assay of PI3K-γ material [γ- 33[P]ATP (10 mCi / mL) and wheat germ agglutinin (WGA) YSi SPA scintillation beads were purchased from Perkin-Elmer (Waltham, MA). The lipid kinase substrate, D-myo-phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) D(+)-sn-1,2-di-O-octanoylglyceryl, 3-O-phospholinked (PIP2), CAS 204858-53-7, was purchased from Echelon Biosciences (Salt Lake City, UT). PI3Kγ (p110γ) recombinant human protein was purchased from Life Technologies (Grand Island, NY). ATP, MgCl2, DTT, EDTA, MOPS, and CHAPS were purchased from Sigma-Aldrich (St. Louis, MO).

[0258] Kinase reactions were performed in a final volume of 25 μL in polystyrene 384-well Greiner Bio-one white plates (Thermo Fisher Scientific). Inhibitors were first serially diluted in DMSO and added to the plate wells before the addition of other reaction components. The final concentration of DMSO in the assay was 2%. PI3Kγ assays were performed at room temperature in 20 mM MOPS (pH 6.7), 10 mM MgCl2, 5 mM DTT, and 0.03% CHAPS. Reactions were initiated by the addition of ATP, and the final reaction mixture contained 20 μM PIP2, 2 μM ATP, 0.5 μCi of [γ- 33 The reaction consisted of 13 nM PI3Kγ, 163 mM potassium phosphate (pH 7.8), 20% glycerol, and 25 mM EDTA. The final concentration of SPA beads was 1.0 mg / mL. After sealing, the plates were shaken overnight at room temperature and centrifuged at 1500 rpm for 10 minutes. The radioactivity of the products was determined by scintillation counting on a Topcount (Perkin-Elmer). IC 50Determination of was performed by curve fitting of percent activity of solvent control against the logarithm of inhibitor concentration using GraphPad Prism 6.0 software.

[0259] Example C. Scintillation Proximity Assay of PI3Kδ material [γ- 33 [P]ATP (10 mCi / mL) and wheat germ agglutinin (WGA) YSi SPA scintillation beads were purchased from Perkin-Elmer (Waltham, MA). The lipid kinase substrate, D-myo-phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) D(+)-sn-1,2-di-O-octanoylglyceryl, 3-O-phospholinked (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, MgCl2, DTT, EDTA, MOPS, and CHAPS were purchased from Sigma-Aldrich (St. Louis, MO).

[0260] Kinase reactions were performed in a final volume of 25 μL in polystyrene 384-well Greiner Bio-one white plates (Thermo Fisher Scientific). Inhibitors were first serially diluted in DMSO and added to the plate wells before the addition of other reaction components. The final concentration of DMSO in the assay was 2%. PI3Kδ assays were performed at room temperature in 20 mM MOPS (pH 6.7), 10 mM MgCl2, 5 mM DTT, and 0.03% CHAPS. Reactions were initiated by the addition of ATP, and the final reaction mixture contained 20 μM PIP2, 2 μM ATP, 0.5 μCi of [γ- 33The reaction consisted of 3.4 nM PI3Kδ, 3.4 nM [P]ATP, and 3.4 nM PI3Kδ. Reactions were incubated for 120 min and terminated by the addition of 40 μL of SPA beads suspended in a stop buffer of 163 mM potassium phosphate (pH 7.8), 20% glycerol, and 25 mM EDTA. The final concentration of SPA beads was 1.0 mg / mL. After sealing the plates, they 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 50 Determination of was performed by curve fitting of percent activity of solvent control against the logarithm of inhibitor concentration using GraphPad Prism 6.0 software.

[0261] The compounds of Examples 1, 5, 6 and 9 were tested in the assays described in Examples A, B, and C and had IC values ​​as shown in Table A below. 50 It was found to have value. [Table 10] + IC ≤ 100 nM 50 ++ indicates IC ≤ 500 nM 50 +++ indicates an IC<2000nM 50 ++++ indicates an IC of ≥ 2000 nM 50 Refers to... # indicates IC ≤ 100 nM 50 ## indicates IC ≤ 500nM 50 indicates an IC of <1000nM 50 #### indicates an IC of ≥ 1000 nM 50 Refers to...

[0262] Various modifications of the present invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. 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 publications, is hereby incorporated 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-hydroxypropanamide.

2. 2. The crystalline form of claim 1, which is anhydrous and unsolvated.

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

4. 4. The crystalline form of any one of claims 1 to 3, having an X-ray powder diffraction pattern comprising at least one peak at 8.6°±0.2° in terms of 2θ.

5. 5. The crystalline form of any one of claims 1 to 4, having an X-ray powder diffraction pattern comprising at least one peak at 9.5°±0.2° in terms of 2θ.

6. 6. The crystalline form of any one of claims 1 to 5, having an X-ray powder diffraction pattern comprising the following peaks at 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°.

7. 7. The crystalline form of any one of claims 1 to 6, having an X-ray powder diffraction pattern containing four or more of the following peaks 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°.

8. 8. The crystalline form of any one of claims 1 to 7, having an X-ray powder diffraction pattern substantially as shown in Figure 1.

9. 9. The crystalline form of any one of claims 1 to 8, having a DSC thermogram comprising an endothermic peak having a maximum at about 193°C.

10. 10. The crystalline form of any one of claims 1 to 9, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 2.

11. 11. The crystalline form of any one of claims 1 to 10, having a thermogravimetric analysis (TGA) substantially as shown in Figure 3.

12. 3. The crystalline form of claim 1 or 2, which is form IIA.

13. 13. The crystalline form of any one of claims 1, 2, and 12, having an X-ray powder diffraction pattern comprising at least one peak at 9.1°±0.2° in terms of 2θ.

14. 14. The crystalline form of any one of claims 1, 2, 12, and 13, having an X-ray powder diffraction pattern comprising at least one peak at 11.1°±0.2° in terms of 2θ.

15. 15. The crystalline form of any one of claims 1, 2, and 12-14, having an X-ray powder diffraction pattern comprising at least one peak at 12.6°±0.2° in terms of 2θ.

16. 16. The crystalline form of any one of claims 1, 2, and 12-15, having an X-ray powder diffraction pattern comprising at least one peak at 13.5°±0.2° in terms of 2θ.

17. 17. The crystalline form of any one of claims 1, 2, and 12-16, having an X-ray powder diffraction pattern comprising the following peaks at 2θ: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; and 13.5°±0.2°.

18. 18. The crystalline form of any one of claims 1, 2, and 12-17, having an X-ray powder diffraction pattern containing four or more of the following peaks 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°.

19. 19. The crystalline form of any one of claims 1, 2, and 12-18, having an X-ray powder diffraction pattern substantially as shown in Figure 4.

20. 20. The crystalline form of any one of claims 1, 2, and 12-19, having a DSC thermogram comprising an endothermic peak having a maximum at about 180°C.

21. 21. The crystalline form of any one of claims 1, 2, and 12-20, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 5.

22. 22. The crystalline form of any one of claims 1, 2, and 12-21, having a thermogravimetric analysis (TGA) substantially as shown in Figure 6.

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

24. 24. The crystalline form of any one of claims 1, 2, and 23 having an X-ray powder diffraction pattern comprising at least one peak at 8.1°±0.2° in terms of 2θ.

25. 25. The crystalline form of any one of claims 1, 2, 23, and 24, having an X-ray powder diffraction pattern comprising at least one peak at 10.6°±0.2° in terms of 2θ.

26. 26. The crystalline form of any one of claims 1, 2, and 23-25, having an X-ray powder diffraction pattern comprising at least one peak at 13.5°±0.2° in terms of 2θ.

27. 27. The crystalline form of any one of claims 1, 2, and 23-26, having an X-ray powder diffraction pattern comprising the following peaks at 2θ: 8.1°±0.2°; 10.6°±0.2°; 13.5°±0.2°; and 14.2°±0.2°.

28. 28. The crystalline form of any one of claims 1, 2, and 23-27, having an X-ray powder diffraction pattern containing four or more of the following peaks 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°.

29. 30. The crystalline form of any one of claims 1, 2, and 23-28, having an X-ray powder diffraction pattern substantially as shown in Figure 7.

30. 30. The crystalline form of any one of claims 1, 2, and 23-29, having a DSC thermogram comprising an endothermic peak having a maximum at about 143°C.

31. 31. The crystalline form of any one of claims 1, 2, and 23-30, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 8.

32. 32. The crystalline form of any one of claims 1, 2, and 23-31, having a thermogravimetric analysis (TGA) substantially as shown in Figure 9.

33. 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.

34. 34. The crystalline form of claim 33, which is anhydrous and unsolvated.

35. 35. The crystalline form of claim 33 or 34, which is form IB.

36. 36. The crystalline form of any one of claims 33 to 35, having an X-ray powder diffraction pattern comprising at least one peak at 6.2°±0.2° in terms of 2θ.

37. 37. The crystalline form of any one of claims 33 to 36, having an X-ray powder diffraction pattern comprising at least one peak at 15.6°±0.2° in terms of 2θ.

38. 38. The crystalline form of any one of claims 33 to 37, having an X-ray powder diffraction pattern comprising the following peaks at 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°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°.

39. 38. The crystalline form of any one of claims 33 to 37, having an X-ray powder diffraction pattern comprising four or more of the following peaks 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°.

40. 40. The crystalline form of any one of claims 33 to 39, having an X-ray powder diffraction pattern substantially as shown in Figure 10.

41. 41. The crystalline form of any one of claims 33 to 40, having a DSC thermogram comprising an endothermic peak having a maximum at about 174°C.

42. 42. The crystalline form of any one of claims 33 to 41, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 11.

43. 35. The crystalline form of claim 33 or 34, which is form IIB.

44. 44. The crystalline form of any one of claims 33, 34 and 43 having an X-ray powder diffraction pattern comprising at least one peak at 4.2°±0.2° in terms of 2θ.

45. 45. The crystalline form of any one of claims 33, 34, 43, and 44, having an X-ray powder diffraction pattern comprising at least one peak at 13.3°±0.2° in terms of 2θ.

46. 46. ​​The crystalline form of any one of claims 33, 34, and 43-45, having an X-ray powder diffraction pattern comprising at least one peak at 17.0°±0.2° in terms of 2θ.

47. 47. The crystalline form of any one of claims 33, 34, and 43-46, having an X-ray powder diffraction pattern comprising at least one peak at 18.8°±0.2° in terms of 2θ.

48. 48. The crystalline form of any one of claims 33, 34, and 43-47, having an X-ray powder diffraction pattern comprising the following peaks at 2θ: 4.3°±0.2°; 7.4°±0.2°; 13.3°±0.2°; and 15.3°±0.2°.

49. 48. The crystalline form of any one of claims 33, 34, and 43-47, having an X-ray powder diffraction pattern containing four or more of the following peaks 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°.

50. 50. The crystalline form of any one of claims 33, 34, and 43-49, having an X-ray powder diffraction pattern substantially as shown in Figure 12.

51. 51. The crystalline form of any one of claims 33, 34, and 43-50, having a DSC thermogram comprising an endothermic peak having a maximum at about 165°C.

52. 51. The crystalline form of any one of claims 33, 34, and 43-50, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 13.

53. Compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ３ A crystalline form of 1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide.

54. 54. The crystalline form of claim 53, which is anhydrous and unsolvated.

55. 55. The crystalline form of claim 53 or 54, which is form IC.

56. 56. The crystalline form of any one of claims 53 to 55, having an X-ray powder diffraction pattern comprising at least one peak at 6.2°±0.2° in terms of 2θ.

57. 57. The crystalline form of any one of claims 53 to 56, having an X-ray powder diffraction pattern comprising at least one peak at 11.9°±0.2° in terms of 2θ.

58. 58. The crystalline form of any one of claims 53 to 57, having an X-ray powder diffraction pattern comprising the following peaks at 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.3°±0.2°; 11.9°±0.2°; and 12.5°±0.2°.

59. 59. The crystalline form of any one of claims 53 to 58, having an X-ray powder diffraction pattern comprising four or more of the following peaks 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°.

60. 60. The crystalline form of any one of claims 53 to 59, having an X-ray powder diffraction pattern substantially as shown in Figure 14.

61. 61. The crystalline form of any one of claims 53 to 60, having a DSC thermogram comprising an endothermic peak having a maximum at about 179°C.

62. 62. The crystalline form of any one of claims 53 to 61, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 15.

63. 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.

64. 64. The salt of claim 63 which is 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide in a 1:1 stoichiometric ratio to hydrobromic acid.

65. 65. The salt of claim 63 or 64, which is substantially isolated.

66. 65. The salt of claim 63 or 64, in a crystalline form.

67. 67. The crystalline form of claim 66, which is a solvated crystalline form.

68. 68. The crystalline form of claim 67, which is a methanol solvate crystalline form.

69. 70. The crystalline form of any one of claims 1 to 62, 67 and 68, which is substantially isolated.

70. 67. A composition comprising a crystalline form according to any one of claims 1 to 62 and 67 to 69, or a salt according to any one of claims 63 to 66.

71. 71. The composition of claim 70, further comprising at least one pharma- ceutically acceptable carrier.

72. A method for inhibiting the activity of PI3Kγ kinase, comprising contacting the kinase with a crystalline form described in any one of claims 1 to 62 and 67 to 69, or a salt described in any one of claims 63 to 66.

73. 73. The method of claim 72, wherein the crystalline form is a selective inhibitor of PI3Kγ over one or more of PI3Kα, PI3Kβ, and PI3Kδ.

74. A method for treating a disease or disorder associated with abnormal expression or activity of PI3Kγ kinase in a patient, the method comprising administering to the patient a therapeutically effective amount of a crystalline form described in any one of claims 1 to 62 and 67 to 69 or a salt described in any one of claims 63 to 66.

75. 75. The method of claim 74, wherein the disease or disorder is an autoimmune disease or disorder, cancer, a cardiovascular disease, or a neurodegenerative disease.

76. 75. The method of claim 74, wherein the disease or disorder is lung cancer, melanoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, colon cancer, endometrial cancer, bladder cancer, skin cancer, uterine cancer, renal cancer, gastric cancer, seminoma, teratocarcinoma, astrocytoma, neuroblastoma, glioma, or sarcoma.

77. 77. The method of claim 76, wherein the sarcoma is Askin tumor, botryoid sarcoma, chondrosarcoma, Ewing sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft part sarcoma, angiosarcoma, cystosarcoma phyllodes, dermatofibrosarcoma protuberans, desmoid tumor, desmoplastic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, angiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma, or undifferentiated pleomorphic sarcoma.

78. 75. The method of claim 74, wherein the disease or disorder is acute myeloid leukemia, acute monocytic leukemia, small lymphocytic lymphoma, chronic lymphocytic leukemia (CLL), chronic myelogenous 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.

79. 79. The method of claim 78, wherein the mature (peripheral) T cell neoplasm (PTCL) is T cell prolymphocytic leukemia, T cell granular lymphocytic leukemia, aggressive NK cell leukemia, mycosis fungoides / Sézary syndrome, anaplastic large cell lymphoma (T cell type), enteropathic T cell lymphoma, adult T cell leukemia / lymphoma, or angioimmunoblastic T cell lymphoma.

80. 79. The method of claim 78, wherein the anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL.

81. The disease or disorder may be Burkitt's lymphoma, acute myeloblastic leukemia, chronic myelogenous 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, 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, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of undetermined significance (MGUS), or diffuse large B-cell lymphoma.

82. 82. The method of claim 81, wherein the non-Hodgkin's lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, indolent NHL (iNHL), or aggressive NHL (aNHL).

83. 82. The method of claim 81, 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.

84. 82. The method of claim 81, wherein the Burkitt lymphoma is endemic Burkitt lymphoma, sporadic Burkitt lymphoma, or Burkitt-like lymphoma.

85. 85. The method of claim 84, wherein the disease or disorder is rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, asthma, allergy, allergic rhinitis, pancreatitis, psoriasis, anaphylaxis, glomerulonephritis, inflammatory bowel disease, thrombosis, meningitis, encephalitis, diabetic retinopathy, benign prostatic hyperplasia, myasthenia gravis, Sjogren's syndrome, osteoarthritis, restenosis, or atherosclerosis.

86. 85. The method of claim 84, wherein 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, bacterial infection, viral infection, graft 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), autoimmune hemolytic anemia, vasculitis, systemic lupus erythematosus, lupus nephritis, pemphigus, or membranous nephropathy.

87. 87. The method of claim 86, wherein the idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP.

88. 87. The method of claim 86, wherein the vasculitis is Behcet's 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 (essential or hepatitis C virus (HCV)-induced), Henoch-Schonlein purpura (HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener's granulomatosis, or antineutrophil cytoplasmic antibody-associated (ANCA) systemic vasculitis (AASV).

89. 87. The method of claim 86, wherein the disease or disorder is Alzheimer's disease, central nervous system injury, or stroke.

90. 1. A process 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, comprising dissolving the compound in a solvent to form a mixture and crystallizing the compound from the mixture.

91. 91. The process of claim 90, wherein the solvent comprises isopropyl acetate.

92. 92. The process of claim 91, wherein the solvent further comprises heptane.

93. 93. The process of any one of claims 90 to 92, further comprising heating the mixture to a temperature of about 70°C to about 90°C.

94. 94. The process of claim 93, further comprising cooling the mixture to room temperature.

95. 91. The process of claim 90, wherein the solvent comprises methanol.

96. 91. The process of claim 90, further comprising heating the mixture to a temperature of about 50° C. to about 70° C.

97. 97. The process of claim 96, further comprising cooling the mixture to room temperature.

98. 98. 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 prepared according to the process of any one of claims 90 to 97.

99. 99. The crystalline form of claim 98, which is anhydrous and unsolvated.

100. 100. The crystalline form of claim 98 or 99, which is form IA.

101. 100. The crystalline form of claim 98 or 99, which is form IIA.

102. 100. The crystalline form of claim 98 or 99, which is Form IIIA.

103. 1. A process for 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, comprising dissolving the compound in a solvent to form a mixture and crystallizing the compound from the mixture.

104. 104. The process of claim 103, further comprising heating the mixture to a temperature of about 70°C to about 90°C.

105. 104. The process of claim 103, further comprising cooling the mixture to room temperature.

106. 106. The process of any one of claims 103 to 105, wherein the solvent comprises isopropyl acetate.

107. 107. The process of claim 106, wherein the solvent further comprises heptane.

108. 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 prepared according to the process of any one of claims 103 to 107.

109. 109. The crystalline form of claim 108, which is anhydrous and unsolvated.

110. 110. The crystalline form of claim 108 or 109, which is form IB.

111. 110. The crystalline form of claim 108 or 109, which is form IIB.

112. Compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ３ 20. A process for preparing a crystalline form of 1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, comprising dissolving the compound in a solvent to form a mixture and crystallizing the compound from the mixture.

113. 113. The process of claim 112, further comprising heating the mixture to a temperature of about 70°C to about 90°C.

114. 113. The process of claim 112, further comprising cooling the mixture to room temperature.

115. 115. The process of any one of claims 112 to 114, wherein the solvent comprises isopropyl acetate.

116. 116. The process of claim 115, wherein the solvent further comprises heptane.

117. The compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d)) prepared according to the process of any one of claims 112 to 116. ３ A crystalline form of 1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide.

118. 118. The crystalline form of claim 117, which is anhydrous and unsolvated.

119. 119. The crystalline form of claim 117 or 118, which is form IC.

120. 1. A process for preparing the hydrobromide salt of a compound, 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, comprising dissolving the compound in a solvent to form a mixture and adding hydrobromic acid to the mixture.

121. 121. The process of claim 120, wherein the solvent comprises methanol.

122. 122. The process of claim 120 or 121, wherein the hydrobromic acid is added to the mixture as an aqueous solution of hydrobromic acid.

123. 123. The process of any one of claims 120 to 122, wherein an excess of hydrobromic acid based on one equivalent of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide is added to the mixture.

124. 124. The process of any one of claims 120 to 123, wherein about 1.1 to about 1.5 equivalents of hydrobromic acid based on 1 equivalent of said 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide is added to the mixture.

125. 125. The process of any one of claims 120 to 124, 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.

126. 126. The process of claim 125, wherein the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide is isolated in a crystalline form.

127. 127. The process of claim 125 or 126, wherein the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide is isolated as a methanol solvate crystalline form.

128. 128. The hydrobromide salt of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide prepared according to the process of any one of claims 120 to 127.

129. 129. The hydrobromide salt of claim 128, which is crystalline.

130. 130. The hydrobromide salt of claim 129, in a solvated crystalline form.

131. 131. The hydrobromide salt of claim 129 or 130, in a methanol solvate crystalline form.

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