Crystalline morphology of phosphoinositide 3-kinase (PI3K) inhibitors

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 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-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 salts and crystalline forms 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, which are PI3K inhibitors useful for the treatment of cancer and other diseases. Background of the Invention

[0002] Compounds: 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-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] This is a phosphoinositide 3-kinase (PI3K) inhibitor useful for treating various diseases, including cancer. For drug development, it is generally advantageous to utilize drug forms that possess desirable properties regarding preparation, purification, reproducibility, stability, bioavailability, and other characteristics. Therefore, the salts and crystalline forms of the compounds of formulas A, B, and C presented herein address the ongoing need for the development of PI3K inhibitors for the treatment of serious diseases. [Overview of the Initiative]

[0003] The present invention relates to the salt and crystalline form of the compound of formula A, the crystalline form of the compound of formula B, and the crystalline form of the 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 shown herein, and at least one pharmaceutically acceptable carrier. The present invention further provides compositions comprising the crystalline form of the compound of formula B and at least one pharmaceutically acceptable carrier. The present invention further provides compositions comprising the crystalline form of the compound of formula C and at least one pharmaceutically acceptable carrier.

[0005] The present invention further provides a process for preparing the salt and crystalline form of the present invention.

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

[0007] [Figure 1]The XRPD pattern for crystalline form IA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide is shown. [Figure 2] The results of DSC experiments on the crystal form IA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide are shown. [Figure 3] The results of TGA experiments on the crystal form IA of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide are shown. [Figure 4] The XRPD pattern for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide in crystalline form IIA is shown. [Figure 5] The results of DSC experiments on 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystal form IIA, are shown. [Figure 6] The results of the TGA experiment for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystal form IIA, are shown. [Figure 7] The XRPD pattern for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide in crystalline form IIIA is shown. [Figure 8]The results of DSC experiments on 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIIA, are shown. [Figure 9] The results of the TGA experiment for 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystal form IIIA, are shown. [Figure 10] 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 is shown for its crystalline form IB. [Figure 11] The results of DSC experiments 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, are shown. [Figure 12] 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 in crystalline form IIB is shown. [Figure 13] The results of DSC experiments 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, are shown. [Figure 14] 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 shows the XRPD pattern for its crystalline form IC. [Figure 15] The results of DSC experiments regarding the crystalline form IC 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 are shown. [Figure 16A] The asymmetric crystalline unit of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide, methanol solvate form, is shown using a thermal ellipsoid drawn at a 30% probability level. [Figure 16B] The crystalline units of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide are shown using a thermal ellipsoid drawn at a 30% probability level. [Modes for carrying out the invention]

[0008] The present invention relates, in particular, to the salt and crystalline form of the PI3K inhibitor 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide having the following formula A, [ka] This is useful, for example, for preparing solid dosage forms of the above compounds for the treatment of various diseases, including cancer.

[0009] The present invention also relates, in particular, to the 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 the following formula B. [ka]

[0010] The present invention also relates, in particular, to the 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 the following formula C. [ka]

[0011] Generally, different crystalline forms of the same substance have different bulk properties, such as hygroscopicity, solubility, and stability. Forms with high melting points often have good thermodynamic stability, which is advantageous for extending the shelf life of drug formulations containing solids. Forms with low melting points are often less thermodynamically stable, but they are advantageous in that they have high water solubility and thus higher bioavailability of the drug. Forms with low hygroscopicity are desirable for their stability to heat and humidity and for their ability to withstand decomposition during long-term storage. Anhydrous forms are often desirable because they can be produced consistently 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 because they are less likely to be hygroscopic and may exhibit improved stability to humidity under storage conditions.

[0012] As used herein, the term “crystal morphology” refers to a particular lattice configuration of a crystalline material. Different crystal morphologies of the same material generally have different crystal lattices (e.g., unit cells), which result from different physical properties specific to each crystal morphology. In some cases, different lattice configurations have different water or solvent content. Different crystal lattices can be identified by solid-state characterization methods, for example, by X-ray powder diffraction (XRPD). Other characterization methods, such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), and solid-state NMR, further assist in the identification of crystal morphology and, similarly, in the determination of stability and solvent / water content.

[0013] The crystalline form of a substance includes both solvated (e.g., hydrated) and solvated (e.g., anhydrous) forms. The hydrated form is a crystalline form in which water is present in the crystal lattice. The hydrated form may also be a stoichiometric hydrate in which water exists in a specific water / molecular ratio, such as a hemihydrate, monohydrate, or dihydrate, within the lattice. The hydrated form may also be non-stoichiometric, where the water content is variable and is affected by external conditions such as humidity.

[0014] Crystal morphology is most commonly characterized by XRPD. The XRPD pattern of reflections (peaks) is typically considered a fingerprint of a particular crystal morphology. It is well known that the relative intensity of XRPD peaks can vary widely depending, among other things, on sample preparation techniques, crystal size distribution, filters, sample mounting procedures, and the specific instrument used. In some cases, new peaks may be observed or existing peaks may disappear depending on the type or setting of the instrument (e.g., whether or not a Ni filter is used). As used herein, the term “peak” refers to a reflection with a relative height / intensity of at least about 4% of the maximum peak height / intensity. Furthermore, changes in instrumentation and other factors may affect the 2-theta value. 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 means encompassing the aforementioned variations.

[0015] Similarly, temperature measurements for DSC, TGA, or other thermal experiments can vary by approximately ±4°C depending on the instrument, specific settings, sample preparation, etc. For example, with respect to DSC, the observed temperature is known to depend on the rate of temperature change, the sample preparation technique used, and the specific instrument. Therefore, the values ​​reported herein for DSC thermograms can vary by as much as ±4°C, as described above. Accordingly, it is understood that the crystal morphologies reported herein having DSC thermograms that are "substantially" as shown in any of the figures correspond to such variations.

[0016] Crystalline form of compound A Compounds of formula A can be isolated in a number of crystalline forms, including, for example, anhydrous and / or solvated or solvated crystalline forms. In some embodiments, the crystalline form of compound A is solvated. In some embodiments, the crystalline form of compound A is anhydrous. In some embodiments, the crystalline form of compound A is solvated. In some embodiments, the crystalline form of compound A is anhydrous and solvated. "Anhydrous" means that the crystalline form of compound A essentially does not contain bound water in its crystalline lattice structure, i.e., the compound does not form crystalline hydrates.

[0017] In some embodiments, the present application provides a process for preparing the crystalline form of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide. In some embodiments, the process comprises dissolving 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-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 includes methanol.

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

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

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

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

[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]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide. In some embodiments, the process comprises dissolving the 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 hydrobromide salts, the solvent includes methanol.

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

[0027] In some embodiments of the process for preparing hydrobromide, an excess amount of hydrobromic acid is added to the mixture based on 1 equivalent of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

[0028] In some embodiments of the process for preparing hydrobromide, 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]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

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

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

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

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

[0033] In some embodiments, the crystalline forms of the present invention are substantially isolated. "Substantially isolated" means 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 impurities of 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%. The impurities generally include, for example, other crystalline forms and other substances that are not substantially isolated crystalline forms.

[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 includes a specific crystalline form in greater than about 80% by weight, greater than about 90% by weight, greater than about 95% by weight, greater than about 98% by weight, greater than about 99% by weight, or greater than about 99.5% by weight.

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

[0036] The crystalline morphology IA of the compound of formula A can be identified, for example, by its unique signature with respect to X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, the crystalline morphology 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, the crystalline form IA of the compound of formula A is characterized by an XRPD pattern including a peak at 8.6°±0.2° at 2θ. In some embodiments, the crystalline form IA of the compound of formula A is characterized by an XRPD pattern including a peak at 9.5°±0.2° at 2θ. In some embodiments, the crystalline form IA of the compound of formula A is characterized by an XRPD pattern including a peak at 10.3°±0.2° at 2θ. In some embodiments, the crystalline form IA of the compound of formula A is characterized by an XRPD pattern including a peak at 14.9°±0.2° at 2θ.

[0038] In some embodiments, the crystalline form IA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more 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°; 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, the crystalline form IA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the crystalline form IA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, morphology IA of the compound of formula A is characterized by a DSC thermogram containing an endothermic peak having a maximum at approximately 193°C. In some embodiments, crystalline morphology IA of the compound of formula A has a DSC thermogram substantially as shown in Figure 2.

[0041] In some embodiments, the crystalline form IA of the compound of formula A has a TGA trace substantially as shown in Figure 3.

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

[0043] The crystalline form IIA of the compound of formula A can be identified, for example, by its unique signature with respect to X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, the crystalline form IIA of the compound of formula A is 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, the crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak at 9.1°±0.2° at 2θ. In some embodiments, the crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak at 11.1°±0.2° at 2θ. In some embodiments, the crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak at 21.9°±0.2° at 2θ. In some embodiments, the crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak at 12.6°±0.2° at 2θ. In some embodiments, the crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak at 13.6°±0.2° at 2θ. In some embodiments, the crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak at 18.0°±0.2° at 2θ. In some embodiments, the crystalline form IIA of the compound of formula A is characterized by an XRPD pattern that includes a peak at 19.0°±0.2° in 2θ.

[0045] In some embodiments, the crystalline form IIA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; and 13.5°±0.2°. In some embodiments, the crystalline form IIA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IIA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 12.6°±0.2°; 18.0°±0.2°; 19.0°±0.2°; and 21.9°±0.2°.

[0046] In some embodiments, the crystalline form IIA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 2θ: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; 13.5°±0.2°; 18.0°±0.2°; 19.0°±0.2°; 20.5°±0.2°; and 21.9°±0.2°. In some embodiments, the crystalline form IIA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the crystalline form IIA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, morphology IIA of the compound of formula A is characterized by a DSC thermogram containing an endothermic peak having a maximum at approximately 180°C. In some embodiments, crystalline morphology IIA of the compound of formula A has a DSC thermogram substantially as shown in Figure 5.

[0048] In some embodiments, the crystalline form IIA of the compound of formula A has a TGA trace substantially as shown in Figure 6.

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

[0050] The crystalline form IIIA of the compound of formula A can be identified, for example, by its unique signature with respect to X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In some embodiments, the crystalline form IIIA of the compound of formula A is 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, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern including a peak at 8.1°±0.2° at 2θ. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern including a peak at 10.6°±0.2° at 2θ. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern including a peak at 13.5°±0.2° at 2θ. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern including a peak at 14.2°±0.2° at 2θ. In some embodiments, the crystalline form IIIA of the compound of formula A is characterized by an XRPD pattern including a peak at 20.3°±0.2° at 2θ.

[0052] In some embodiments, the crystalline form IIIA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 8.1°±0.2°; 10.6°±0.2°; 13.5°±0.2°; and 14.2°±0.2°. In some embodiments, the crystalline form IIIA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IIIA of the compound of formula A has an XRPD pattern at 2θ that includes the following peaks: 10.6°±0.2°; 13.5°±0.2°; 14.2°±0.2°; and 20.3°±0.2°.

[0053] In some embodiments, the crystalline form IIIA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the crystalline form IIIA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the crystalline form IIIA of the compound of formula A has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, morph IIIA of the compound of formula A is characterized by a DSC thermogram containing an endothermic peak having a maximum at approximately 143°C. In some embodiments, crystalline morph IIIA of the compound of formula A has a DSC thermogram substantially as shown in Figure 8.

[0055] In some embodiments, the crystalline form IIIA of the compound of formula A has a TGA trace substantially as shown in Figure 9.

[0056] Hydrobromide of formula A In some embodiments, this application provides hydrobromide salts of the compound of formula A. In some embodiments, the hydrobromide salt of the compound of formula A is 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide in a 1:1 stoichiometric ratio with respect to hydrobromic acid. The hydrobromide salt form can generally be prepared 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 solvated crystalline form. In some embodiments, the hydrobromide salt of the compound of formula A is in methanol solvate crystalline form.

[0058] Crystalline form of compound B Compounds of formula B can be isolated in a number of crystalline forms, including, for example, anhydrous and / or solvated crystalline forms. In some embodiments, the crystalline form of compound B is anhydrous. In some embodiments, the crystalline form of compound B is solvated. In some embodiments, the crystalline forms of compound B are anhydrous and solvated. "Anhydrous" means that the crystalline form of compound B essentially does not contain bound water in its crystalline lattice structure, i.e., the compound does not form crystalline hydrates.

[0059] In some embodiments, the crystalline forms shown 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 then crystallizing the compound from the mixture.

[0060] In some embodiments, the process further includes heating the mixture to a temperature of about 70°C to about 90°C. In some embodiments, the process further includes 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, this 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 process described 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. "Substantially isolated" means 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 impurities of 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%. The impurities generally include, for example, other crystalline forms and other substances that are not substantially isolated crystalline forms.

[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 includes a specific crystalline form in greater than about 80% by weight, greater than about 90% by weight, greater than about 95% by weight, greater than about 98% by weight, greater than about 99% by weight, or greater than about 99.5% by weight.

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

[0066] The crystalline form IB of the compound of formula B can be identified, for example, by its unique signature with respect to X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the 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, the crystalline form IB of the compound of formula B is characterized by an XRPD pattern including a peak at 6.2°±0.2° at 2θ. In some embodiments, the crystalline form IB of the compound of formula B is characterized by an XRPD pattern including a peak at 15.6°±0.2° at 2θ. In some embodiments, the crystalline form IB of the compound of formula B is characterized by an XRPD pattern including a peak at 20.7°±0.2° at 2θ. In some embodiments, the crystalline form IB of the compound of formula B has an XRPD pattern including the following peaks at 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, the crystalline form IB of the compound of formula B has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IB of the compound of formula B has an XRPD pattern at 2θ that includes the following peaks: 6.2°±0.2°; 12.0°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 19.3°±0.2°; 20.7°±0.2°; 23.2°±0.2°; and 27.1°±0.2°. In some embodiments, the crystalline form IB of the compound of formula B has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IB of the compound of formula B has an XRPD pattern containing two or more, three or more, or four or more peaks at 2θ: 2θ: 6.2°±0.2°; 10.4°±0.2°; 11.4°±0.2°; 11.6°±0.2°; 12.0°±0.2°; 13.9°±0.2°; 14.4°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°. In some embodiments, the crystalline form IB of the compound of formula B has an XRPD pattern containing two or more, three or more, or four or more peaks at 2θ: 6.2°±0.2°; 12.0°±0.2°; 15.6°±0.2°; 16.0°±0.2°; 16.7°±0.2°; 19.3°±0.2°; 20.7°±0.2°; 23.2°±0.2°; and 27.1°±0.2°. In some embodiments, the crystalline form IB of the compound of formula B has an XRPD pattern containing two or more, three or more, or four or more 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°; and 14.4°±0.2°.

[0069] In some embodiments, the morphology IB of the compound of formula B is characterized by a DSC thermogram containing an endothermic peak having a maximum at approximately 174°C. In some embodiments, the crystalline morphology IB of the compound of formula B has a DSC thermogram substantially as shown in Figure 11.

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

[0071] The crystalline form IIB of the compound of formula B can be identified, for example, by its unique signature with respect to X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the 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, the crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak at 4.2°±0.2° at 2θ. In some embodiments, the crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak at 7.4°±0.2° at 2θ. In some embodiments, the crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak at 13.3°±0.2° at 2θ. In some embodiments, the crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak at 20.1°±0.2° at 2θ. In some embodiments, the crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak at 17.0°±0.2° at 2θ. In some embodiments, the crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak at 18.8°±0.2° at 2θ.

[0073] In some embodiments, the crystalline form IIB of the compound of formula B has an XRPD pattern at 2θ that includes the following peaks: 4.3°±0.2°; 7.4°±0.2°; 13.3°±0.2°; and 15.3°±0.2°. In some embodiments, the crystalline form IIB of the compound of formula B has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IIB of the compound of formula B has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the crystalline form IIB of the compound of formula B has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, morph IIB of the compound of formula B is characterized by a DSC thermogram containing an endothermic peak having a maximum at approximately 165°C. In some embodiments, crystalline morph IIB of the compound of formula B has a DSC thermogram substantially as shown in Figure 13.

[0076] Crystalline form of compound C Compounds of formula C can be isolated in a number of crystalline forms, including, for example, anhydrous and / or solvated crystalline forms. In some embodiments, the crystalline form of compound C is anhydrous. In some embodiments, the crystalline form of compound C is solvated. In some embodiments, the crystalline forms of compound C are anhydrous and solvated. "Anhydrous" means that the crystalline form of compound C essentially does not contain bound water in its crystalline lattice structure, i.e., the compound does not form crystalline hydrates.

[0077] In some embodiments, the present application provides a process for preparing the 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 comprises 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 includes heating the mixture to a temperature of about 70°C to about 90°C. In some embodiments, the process further includes cooling the mixture to room temperature.

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

[0080] This 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 process described 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. "Substantially isolated" means 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 impurities of 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%. The impurities generally include, for example, other crystalline forms and other substances that are not substantially isolated crystalline forms.

[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 includes a specific crystalline form in greater than about 80% by weight, greater than about 90% by weight, greater than about 95% by weight, greater than about 98% by weight, greater than about 99% by weight, or greater than about 99.5% by weight.

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

[0084] The crystalline form IC of a compound of formula C can be identified, for example, by its unique signature with respect to X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the crystalline form IC of a 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, the crystalline IC of the compound of formula C is characterized by an XRPD pattern including a peak at 6.2°±0.2° at 2θ. In some embodiments, the crystalline IC of the compound of formula C is characterized by an XRPD pattern including a peak at 11.9°±0.2° at 2θ. In some embodiments, the crystalline IC of the compound of formula C is characterized by an XRPD pattern including a peak at 16.7°±0.2° at 2θ. In some embodiments, the crystalline form IC of the compound of formula C has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IC of the compound of formula C has an XRPD pattern at 2θ that includes the following peaks: 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, the crystalline form IC of the compound of formula C has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the crystalline form IC of the compound of formula C has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the crystalline form IC of the compound of formula C has an XRPD pattern containing two or more, three or more, or four or more peaks at 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, the morphological IC of the compound of formula C is characterized by a DSC thermogram that includes an endothermic peak having a maximum at approximately 179°C. In some embodiments, the crystalline morphological IC of the compound of formula C has a DSC thermogram substantially as shown in Figure 15.

[0088] How to use The compounds of the present invention described herein (e.g., salts and crystalline forms) inhibit the activity of PI3Kγ kinase. Therefore, the salts and crystalline forms of the present invention described herein can be used in methods of inhibiting PI3Kγ kinase by contact with the kinase. In some embodiments, the salts and crystalline forms of the present invention can be used in methods of inhibiting PI3Kγ activity in individuals / patients requiring inhibition by administering an effective amount of the salt or crystalline form described herein. In some embodiments, the modification is inhibition. In some embodiments, the contact is in vivo. In some embodiments, the contact is ex vivo. Advantageously, the crystalline forms described herein exhibit superior efficacy and favorable safety and toxicity profiles in animal studies.

[0089] In some embodiments, PI3Kγ contains mutations. Mutations may be the substitution of one amino acid with another, or the deletion of one or more amino acids. In such embodiments, mutations may be located in the kinase domain of PI3Kγ.

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

[0091] The salts and crystalline forms described herein may be selective. “Selective” means that the salt or crystalline form binds to or inhibits PI3Kγ with greater affinity or potency than at least one other kinase. In some embodiments, the salts and crystalline forms of the disclosure are selective inhibitors of PI3Kγ more than PI3Kδ, PI3Kα, and PI3Kβ. In some embodiments, the selectivity may be at least about 2-fold, 3-fold, 5-fold, 10-fold, or 20-fold higher than PI3Kδ when measured by the assays described herein. In some embodiments, the selectivity can be tested at a 2 μM ATP concentration for each enzyme. In some embodiments, the selectivity of the salts and crystalline forms of the disclosure can be determined by cell assays related to specific PI3K kinase activity.

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

[0093] In some embodiments, the disease or disorder is an autoimmune disease or disorder, cancer, cardiovascular disease, or 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, staphyloid sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft part sarcoma, angiosarcoma, phyllodes sarcoma, dermatofibrosarcoma protuberans, desmoid tumor, fibroplastic round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), perivascular cell tumor, 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), lymphocytic lymphoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma, T-cell acute lymphoblastic leukemia (T-ALL), cutaneous T-cell lymphoma, large granular lymphocytic leukemia, mature (peripheral) T-cell neoplasm (PTCL), anaplastic large cell lymphoma (ALCL), or lymphoblastic lymphoma. In some embodiments, mature (peripheral) T-cell neoplasms (PTCLs) are pre-lymphocytic T-cell leukemia, granular lymphocytic T-cell leukemia, invasive NK-cell leukemia, mycosis fungoides / Sézary syndrome, naplastic large cell lymphoma (T-cell type), enteropathy-type T-cell lymphoma, adult T-cell leukemia / lymphoma, or angioimmunoblastic T-cell lymphoma. In some embodiments, anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL.

[0097] In some embodiments, the disease or disorder is Burkitt lymphoma, acute myeloblastic leukemia, chronic myeloid leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanocyte, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenström macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic lymphoma These include blastic leukemia, myelofibrosis, mucosal-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary exudative lymphoma, intravascular large B-cell lymphoma, plasmacytosis, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammaglobulinemia of unknown 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 hematopoietic changes in pathological conditions such as chronic infections and cancer. MDSCs are distinguished from other myeloid cell types by their potent immunosuppressive activity rather than immunostimulatory properties. Like other myeloid cells, MDSCs interact with other types of immune cells, including T cells, dendritic cells, macrophages, and natural killer cells, to modulate their function. In some embodiments, for example, the compounds described herein may be used in methods related to cancerous tissue (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 Burkitt lymphoma, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenström macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, These include myelofibrosis, mucosa-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary exudative lymphoma, intravascular large B-cell lymphoma, plasmacytosis, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammaglobulinemia of unknown significance (MGUS), or diffuse large B-cell lymphoma.

[0100] In some embodiments, non-Hodgkin lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, low-grade NHL (iNHL), or aggressive NHL (aNHL).

[0101] In some embodiments, 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, 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, Sjögren's syndrome, osteoarthritis, restenosis, or atherosclerosis.

[0104] In some embodiments, the disease or disorder is cardiac hypertrophy, cardiomyocyte dysfunction, acute coronary syndrome, chronic obstructive pulmonary disease (COPD), chronic bronchitis, hypertension, ischemia, ischemia-reperfusion, vasoconstriction, anemia (e.g., hemolytic anemia, aplastic anemia, or euerythrocytic 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 xenograft, 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, cardiomyocyte dysfunction, chronic obstructive pulmonary disease (COPD), hypertension, ischemia, ischemia-reperfusion, vasoconstriction, anemia (e.g., hemolytic anemia, aplastic anemia, or euerythrocytic anemia), bacterial infection, viral infection, graft rejection, kidney disease, anaphylactic shock fibrosis, skeletal muscle atrophy, skeletal muscle hypertrophy, angiogenesis, sepsis, graft 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, idiopathic thrombocytopenic purpura (ITP) is relapsing ITP or refractory ITP.

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

[0109] This disclosure further provides salts or crystalline forms described herein for use in any of the methods described herein.

[0110] This disclosure further provides the use of the crystalline forms described herein, or pharmaceutically acceptable salts thereof, for the preparation of pharmaceuticals for use in any of the methods described herein.

[0111] As used herein, the term “contact” means bringing the indicated parts together in an in vitro or in vivo system. For example, “contact” PI3K with the salt or crystalline form of the Disclosure includes administering the crystalline form of the Disclosure to an individual or patient, such as a human, having PI3K, and introducing, for example, the salt or crystalline form of the Disclosure into a sample containing a cell preparation or purified preparation containing PI3K.

[0112] Any of the salts and crystalline forms, or embodiments thereof, described herein are considered to have a sufficient pharmacological profile and promising biopharmaceutical properties, such as toxicological profile, metabolic and pharmacokinetic properties, solubility, and permeability. Determining appropriate biopharmaceutical properties is within the scope of the knowledge of those skilled in the art and is understood to be, for example, determining cytotoxicity in cells or inhibition of a given target or channel to determine potential toxicity.

[0113] As used herein, the terms “individual” or “patient” as used interchangeably refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, most preferably humans.

[0114] As used herein, the term “therapeutic dose” refers to the amount of an active salt, crystalline form, or pharmaceutical product that elicits a desired biological or pharmacological response in a tissue, system, animal, individual, or human, as determined by a researcher, veterinarian, physician, or other clinician.

[0115] As used herein, the term “to treat” or “treatment” can mean one or more of the following: (1) inhibiting a disease, for example, inhibiting a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or overall symptoms of a disease, condition, or disorder (i.e., stopping the further progression of the pathology and / or overall symptoms); and (2) improving a disease, for example, improving a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or overall symptoms of a disease, condition, or disorder (i.e., improving the pathology and / or overall symptoms), 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 any of the diseases referred to herein or reducing the risk of developing any of them, for example, in individuals who may be predisposed to a disease, condition, or disorder but have not yet experienced or shown the pathological symptoms or overall manifestations of that disease.

[0117] Combination therapy I. Immune checkpoint therapy In some embodiments, the PI3Kγ inhibitors provided herein can be used in combination with one or more immune checkpoint inhibitors for the treatment of cancer as described herein. In one embodiment, the combination with one or more immune checkpoint inhibitors as described herein may be used for the treatment of melanoma. The salt and crystalline forms of the Disclosure may be used in combination with one or more immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors against 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 stimulating 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 KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors, and TGFR beta inhibitors.

[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 the 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 the immune checkpoint molecule is a PD-1 inhibitor, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), durvalumab (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-PD1 antibody is pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012. In some embodiments, the anti-PD1 antibody is SHR-1210. Other anticancer agents include antibody therapies such as 4-1BB (e.g., urelumab, utomirumab).

[0121] In some embodiments, the inhibitor of the immune checkpoint molecule is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

[0122] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-1 and PD-L1, for example, 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 the immune checkpoint molecule is an inhibitor of CTLA-4, for example, 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 the immune checkpoint molecule is an inhibitor of LAG3, for example, an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, or INCAGN2385.

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

[0127] In some embodiments, the inhibitor of the 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 the 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 the immune checkpoint molecule is a CD20 inhibitor, such as an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

[0130] The salts and crystalline forms of the disclosed herein 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 the TGFβ receptor.

[0131] In some embodiments, the PI3Kγ inhibitors provided herein may be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitors are inhibitors 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, additional compounds, inhibitors, drugs, etc., may be combined with this compound in a single or sequential dosage form, or they may be administered simultaneously or sequentially in separate dosage forms.

[0133] II. Cancer Treatment The proliferation and survival of cancer cells can be influenced by multiple signaling pathways. Therefore, combining different enzyme / protein / receptor inhibitors that exhibit different selectivity at targets that modulate activity is useful for treating such conditions. Targeting two or more signaling pathways (or two or more biological molecules involved in a given signaling pathway) can reduce the potential for drug resistance to develop in a cell population and / or reduce the toxicity of treatment.

[0134] The salts and crystalline forms of the disclosed herein may 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 are listed herein. Examples of cancer include solid tumors and humoral malignancies such as hematological malignancies.

[0135] For example, one or more additional pharmaceuticals such as chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressants, cancer immunotherapy 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 disclosed for the treatment of PI3K-related diseases, disorders, or conditions. One or more additional pharmaceuticals may be administered to the patient simultaneously or sequentially.

[0136] For example, salts and crystalline forms as disclosed herein can be combined with one or more of the following kinase inhibitors for the treatment of cancer and other diseases or disorders described herein: 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 , 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 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), and LSD1 inhibitors (e.g., INCB59872 and INCB6 Examples include 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 extraterminal 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 more selective to JAK2 than to JAK2.

[0138] Examples of antibodies for use in combination therapy include, but are not limited to, antibodies against 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 c-MET.

[0139] One or more of the following drugs may be used in combination with the salts and crystalline forms of the Disclosure, and are presented as a non-limiting list: cell proliferation inhibitors, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epotilon, 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 master Chlormethine, Ifosfamide, Melphalan, Chlorambucil, Pipobroman, Triethylenemelamine, Triethylenethiophosphoramine, Busulfan, Carmustine, Lomustine, Streptozocin, Dacarbazine, Furoxlysine, Cytarabine, 6-Mercaptopurine, 6-Thiogunine, Fludarabine Phosphate, Oxaliplatin, Leucobilin, ELOXATIN (trademark) (Oxaliplatin), Pentostatin, Vinblastine, Vincristine, Vindesine, Bleomycin, Dactinomycin, Daunorubicin, Doxorubicin, Epirubicin, Idarubicin, Mitramycin, Deoxycoformycin, Mitomycin-C, L-Asparaginase, Teniposide 17. Alpha-.- Ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testactone, megstrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianicene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisol, navelben, anastrazole, letrazole, capecitabine, reloxafine, droloxafine, hexamethylmelamine, avastin, HERCEPTIN (trademark) (Trusts Mab), BEXXAR (trademark) (tositumomab), VELCADE (trademark) (bortezomib), ZEVALIN (trademark) (ibritumomab tiuxetan), TRISENOX (trademark) (arsenic trioxide), XELODA (trademark) (capecitabine), vinorelbine, porfimer, ERBITUX (trademark) (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, rerozo Lu, fulvestrant, exemestane, ifosfomide, rituximab, C225 (cetuximab), Campas (aremutuzumab), clofarabine, cladribine, aphidicolon, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triapin, zidox, trimidox, amidox, 3-AP, and MDL-101,731.

[0140] The salts and crystalline forms of this disclosure can be further used in combination with other methods of treating cancer, such as chemotherapy, radiotherapy, 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 virus therapy, and immunomodulatory small molecules, such as thalidomide or JAK1 / 2 inhibitors. The salts and crystalline forms may be administered in combination with one or more anticancer agents, such as chemotherapeutic agents. Examples of chemotherapy drugs include avalerix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezombie, bortezomib, intravenous busulfan, oral busulfan, callussterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, daunorubicin, decitabine, denileukin, denileukin difutitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, and el Rotinib, Estramustine, Etoposide Phosphate, Etoposide, Exemestane, Fentanyl Citrate, Filgrastim, Furoxuridine, Fludarabine, Fluorouracil, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab Ozogamicin, Goserelin Acetate, Histrelin Acetate, Ibritumomab Tiuxetan, Idarubicin, Ifosfamide, Imatinib Mesylate, In Taferon alpha 2a, irinotecan, lapatinib distosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisol, lomustine, mechloretamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone fenpropionate, nelarabine, nofetumomab,This includes any of the following: olapariboxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobromane, pricamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, rucaparib, streptozocin, tamoxifen, temozolomide, teniposide, testactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, barrubicin, vinblastine, vincristine, vinorelbine, vorinostat, niraparib, beriparib, talazoparib, and zoledronate.

[0141] Examples of additional chemotherapy drugs include proteosome inhibitors (e.g., bortezomib), thalidomide, revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, and carmustine.

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

[0143] Examples of Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC®), nilotinib, dasatinib, bosutinib, and ponatinib, as well as pharmaceutically acceptable salts. Other examples of suitable Bcr-Abl inhibitors include compounds of the genera and species disclosed in U.S. Patent No. 5,521,184, WO04 / 005281, and U.S. Patent No. 60 / 578,491, as well as their pharmaceutically acceptable salts.

[0144] Examples of suitable Flt-3 inhibitors include midostaurin, restaurtinib, linifanib, sunitinib, sunitinib maleate, sorafenib, quizartinib, clenolanib, pacritinib, tandutinib, PLX3397 and ASP2215, and their pharmaceutically acceptable salts. Other examples of suitable Flt-3 inhibitors include compounds such as those disclosed in WO03 / 037347, WO03 / 099771 and WO04 / 046120, and their pharmaceutically acceptable salts.

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

[0146] Examples of suitable FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520, and GSK2256098, as well as their pharmaceutically acceptable salts. Other examples of suitable FAK inhibitors include compounds such as those disclosed in WO04 / 080980, WO04 / 056786, WO03 / 024967, WO01 / 064655, WO00 / 053595, and WO01 / 014402, as well as their pharmaceutically acceptable salts.

[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 resistant to imatinib or other kinase inhibitors.

[0148] In some embodiments, the salts and crystalline forms of the Disclosure may be used in combination with chemotherapeutic agents in the treatment of cancer, improving the therapeutic response compared to the response to the chemotherapeutic agent alone without exacerbating its toxic effects. In some embodiments, the salts and crystalline forms of the Disclosure may be used in combination with chemotherapeutic agents provided herein. For example, additional agents used in the treatment of multiple myeloma may include, but are not limited to, melphalan, melphalan + 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 immunomodulator. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulator is lenalidomide (LEN) or pomalidomide (POM). Additive or synergistic effects are desirable outcomes of combining the PI3K inhibitors of this disclosure with additional agents.

[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 drugs may be combined with this compound in a single or sequential dosage form, or the drugs may be administered simultaneously or sequentially in separate dosage forms.

[0151] The salts and crystalline forms of this disclosure may be used in combination with one or more other inhibitors or one or more therapies for the treatment of infections. Examples of infections include viral infections, bacterial infections, fungal infections, or parasitic infections.

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

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

[0154] The salts and crystalline forms of the 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, the tumor vaccine contains proteins derived from viruses involved in human cancer, 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 Disclosure can be used in combination with tumor-specific antigens, such as heat shock proteins isolated from tumor tissue itself. In some embodiments, the salts and crystalline forms can be combined with dendritic cell immunization to activate a potent antitumor response.

[0155] The salts and crystalline forms of the present disclosure can be used in combination with bispecific macrocyclic peptides that direct 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 the host immune response.

[0156] In some further embodiments, combinations of the salts and crystalline forms of the present disclosure with other therapeutic agents can be administered to a patient before, during, and / or after bone marrow transplantation 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 hematopoietic tumors.

[0157] Salts and crystalline forms can be used in combination with vaccines to stimulate an immune response to pathogens, toxins, and autoantigens. Examples of pathogens for which this therapeutic approach may be particularly useful include pathogens 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, leishmania, Staphylococcus aureus, and Pseudomonas aeruginosa.

[0158] Viruses that cause infections treatable by the methods of this disclosure include, but are not limited to, human papillomavirus, influenza, hepatitis A, B, C, or D viruses, 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, Mampus virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, infectious viruses, poliovirus, rabies virus, JC virus, and arboviral encephalitis virus.

[0159] Pathogenic bacteria that cause infections treatable by the methods of this disclosure include, but are not limited to, Chlamydia, Rickettsial bacteria, Mycobacteria, Staphylococci, Streptococci, Pneumonococci, Meningococci and Conococci, Klebsiella, Proteus, Serratia, Pseudodomonas, Legionella, Diphtheria, Salmonella, Bacilli, Cholera, Tetanus, Botulism, Anthrax, Plague, Leptospirosis, and Lyme disease bacteria.

[0160] Pathogenic fungi that cause infections treatable by the methods of this disclosure include, but are not limited to, Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), Genus Mucorales (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 this 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. Furthermore, 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), and its disclosure is incorporated herein by reference as if it were contained in its entirety.

[0162] Pharmaceutical preparations and dosage forms When used as a formulation, the compounds of this disclosure (e.g., in salt and crystalline forms) can be administered in the form of pharmaceutical compositions. These compositions can be prepared in ways well known in the pharmaceutical field and can be administered by various routes depending on whether topical or systemic treatment is desired and the area to be treated. Administration may be topical (including transdermal, epidermal, ocular, and mucosal delivery, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or blowing of powder or aerosol, including by nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration may include intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion, or intracranial, for example, intrathecal or intraventricular administration. Parenteral administration may be in the form of a single bolus dose or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, droplets, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powdered, or oily base materials, thickeners, etc., may be required or desired.

[0163] The Disclosure also includes pharmaceutical compositions comprising, as an active ingredient, one of the compounds of the 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 the preparation of the compositions of the Disclosure, the active ingredient is typically mixed with an excipient, diluted by an excipient, or encapsulated in such a carrier in the form of, for example, a capsule, sachet, paper, or other container. Where the excipient functions as a diluent, it may be a solid, semi-solid, or liquid substance acting as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, sachets, caches, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or liquid media), for example, ointments, soft and hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaging powders containing up to 10% by weight of the active compound.

[0164] When preparing a formulation, the active compound can be ground to an appropriate particle size before being combined with other components. If the active compound is substantially insoluble, it can be ground to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size can be adjusted by grinding to provide a substantially uniform distribution in the formulation, for example, about 40 mesh.

[0165] The compounds of this disclosure may be ground using known grinding procedures, such as wet grinding, to obtain particle sizes suitable for tablet formation and other types of formulations. Finely divided (nanoparticle-like) preparations of the compounds of this disclosure may be prepared by processes known in the art. See, for example, International Application 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 formulations may also include lubricants such as talc, magnesium stearate, and mineral oil; humectants; emulsifiers and suspending agents; preservatives such as methyl benzoate and propyl hydroxybenzoate; sweeteners; and flavoring agents. The compositions of this disclosure may be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to a patient by employing procedures known in the art.

[0167] The composition can be formulated in unit dosage forms, each dose containing approximately 5 to approximately 1000 mg (1 g) of the active ingredient, more typically approximately 100 to approximately 500 mg. The term "unit dosage form" refers to a physically distinct unit suitable as a unitary dosage for human subjects and other mammals, each unit containing a predetermined amount of the active material calculated to produce the desired therapeutic effect in conjunction with a suitable pharmaceutically acceptable excipient.

[0168] In some embodiments, the compositions of the present disclosure contain about 5 to about 50 mg of the active ingredient. Those skilled in the art will understand 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, the compositions of the present disclosure contain about 50 to about 500 mg of the active ingredient. Those skilled in the art will understand that this may embody 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, the compositions of the present disclosure contain about 500 to about 1000 mg of the active ingredient. Those skilled in the art will understand that this may embody 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 this disclosure.

[0172] Active compounds can be effective across a wide range of doses and are generally administered at pharmaceutically effective doses. However, it should be understood that the actual amount of compound administered is usually determined by the physician based on relevant circumstances, including the condition being treated, the chosen route of administration, the actual compound being administered, the individual patient's age, weight, and response, and the severity of the patient's symptoms.

[0173] To prepare solid compositions such as tablets, the main active ingredient is mixed with pharmaceutical excipients to form a solid pre-formulation composition containing a homogeneous mixture of the compounds of this disclosure. When these pre-formulation compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. This solid pre-formulation is then subdivided into, for example, the aforementioned unit dosage forms containing about 0.1 to about 1000 mg of the active ingredient of this disclosure.

[0174] The tablets or pills of this disclosure may be coated or otherwise compounded to provide a dosage form that offers the advantage of a long-lasting effect. For example, the tablets or pills may contain an inner dosage and an outer dosage component, the latter in the form of a coating covering the former. The two components may be separated by an enteric coating that resists disintegration in the stomach and allows the inner component to pass intact into the duodenum or delays its release. A variety of substances may be used for such enteric coatings or coatings, including numerous polymer acids and mixtures of polymer acids with substances such as shellac, cetyl alcohol, and cellulose acetate.

[0175] Liquid forms in which the compounds and compositions of this disclosure may be incorporated for oral administration or by injection include aqueous solutions, preferably flavored syrups, aqueous or oily suspensions, and flavored emulsions having 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 inhalation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, compositions are administered by oral or nasal respiratory pathways for topical or systemic effects. Compositions may be sprayed using an inert gas. The sprayed solution may be inhaled directly from a spraying device, or the spraying device may be attached to a face mask, embolus, or intermittent positive airway pressure (CPAP) respirator. Compositions in solution, suspension, or powder form may be administered orally or nasally from a device that delivers 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. Carrier compositions for creams may be based on water combined with glycerol and one or more other components, for example, glycerin monostearate, PEG-glycerin monostearate, and cetyl stearyl alcohol. Gels can be formulated using isopropyl alcohol and water, preferably in combination 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 the compounds of the present disclosure. Topical formulations may preferably be packaged in, for example, 100 g tubes, optionally associated with instructions for a selected indication, for example, psoriasis or other skin conditions.

[0178] The amount of compound or composition administered to a patient will vary depending on the substance being administered, the purpose of administration (e.g., prevention or treatment), the patient's condition, and the method of administration. For therapeutic purposes, the composition may be administered to patients already suffering from the disease in an amount sufficient to cure or at least partially halt the symptoms of the disease and its complications. The effective dose depends on the judgment of the attending clinician, based on factors such as the state of the disease being treated, the severity of the disease, the patient's age, weight, and general health.

[0179] The compositions administered to patients may be in the form of the pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or aseptically filtered. Aqueous solutions may be packaged or lyophilized for use as is, but lyophilized preparations are combined with a sterile aqueous carrier before administration. The pH of the compound preparation is typically 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. It is understood that the predetermined use of the aforementioned excipients, carriers, or stabilizers results in the formation of a pharmaceutically acceptable salt.

[0180] The therapeutic dose of the compounds disclosed herein may vary depending, for example, the specific use in which the treatment is performed, the method of administering the compound, the patient's health and condition, and the judgment of the prescribing physician. The proportion or concentration of the compounds disclosed herein in a pharmaceutical composition may vary depending on several factors, including the dose, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compounds disclosed herein may be provided in a physiologically buffered aqueous solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dose ranges are about 1 μg / kg body weight to about 1 g / kg body weight per day. In some embodiments, the dose range is about 0.01 mg / kg body weight to about 100 mg / kg body weight per day. The dose may depend on variables such as the type and progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the selected compound, the formulation of excipients, and its route of administration. The effective dose can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0181] The compositions of this disclosure may further include one or more additional pharmaceutical products, 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 this disclosure relates to labeled compounds (e.g., salts and crystalline forms) (radioactively labeled, fluorescently labeled, etc.) of this disclosure that would be useful in assays both in vitro and in vivo, as well as imaging techniques, for locating and quantifying PI3K in tissue samples, including human, and for identifying PI3K ligands, by inhibitory binding of the labeled compounds. Substitution of one or more atoms of the compounds of this disclosure may also be useful in producing differentiated ADME (adsorption, distribution, metabolism, and excretion). Accordingly, this disclosure includes PI3K assays containing such labeled or substituted compounds.

[0183] The present disclosure further includes isotopically-labeled compounds of the present disclosure. An "isotopic" or "radiolabeled" compound is a compound of the present disclosure in which one or more atoms are replaced or substituted by atoms having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated into the compounds of the present disclosure include 2 H (also denoted as D for deuterium), 3 H (also denoted 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 I, but are not limited thereto. For example, one or more hydrogen atoms in a compound of the present disclosure can be replaced by deuterium atoms (e.g., a C of formula (I) 1~6 one or more hydrogen atoms of an alkyl group can be optionally substituted with deuterium atoms such that -CH3 is replaced with -CD3). In some embodiments, all alkyl groups in the formulas and / or forms of the present disclosure may be fully deuterated.

[0184] One or more constituent atoms of the compounds presented herein may be replaced or substituted with isotopes of the atoms in natural or non-natural abundances. In some embodiments, the compound comprises at least one deuterium atom. For example, one or more hydrogen atoms in a compound presented herein can be replaced or substituted by deuterium atoms (e.g., a C 1-6One or more hydrogen atoms of the alkyl group may be replaced by deuterium atoms such that -CH3 is replaced by -CD3. 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 may be replaced or substituted by deuterium atoms.

[0185] In some embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms bonded to the carbon atom of any 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). Isotopic-labeled compounds can be used in various tests such as NMR spectroscopy, metabolic experiments, and / or assays.

[0187] Substitution with heavier isotopes, such as deuterium, may result in certain therapeutic benefits stemming from greater metabolic stability, such as an increased in vivo half-life or a reduced required dose, and may therefore be preferable in some situations. (See, for example, A. Kerekes et al. J. Med. Chem. 2011, 54, 201-210; R. Xu et al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolic sites may result in one or more of these therapeutic benefits.

[0188] The radionuclides incorporated into the radiolabeled compound depend on the specific application of that radiolabeled compound. For example, in the case of in vitro PI3K labeling and competitive assays, 3 H, 14 C, 82 Br, 125 I, 131 I, or 35 Compounds incorporating sulfur may be useful. In imaging applications using radiation, 11 C, 18 F, 125 I, 123 I, 124 I, 131 I, 75 Br, 76 Br, or 77 Br may be useful.

[0189] A “radiolabeled” or “labeled compound” is understood to be a compound incorporating at least one radionuclide. In some embodiments, the radionuclide is: 3 H, 14 C, 125 I, 35 S, and 82 Selected from the group consisting of Br.

[0190] This disclosure may further include synthetic methods for incorporating radioactive isotopes into the compounds of this disclosure. Synthetic methods for incorporating radioactive isotopes into organic compounds are well known in the art, and those skilled in the art will readily recognize methods applicable to the compounds of this disclosure.

[0191] The labeled compounds of this 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 changes when in contact with PI3K through tracking of the label. For example, a test compound (labeled) can be evaluated for its ability to reduce the binding of another compound known to bind to PI3K (i.e., a standard compound). 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 not. Therefore, to evaluate the competition between the standard compound and the test compound, the concentration of the labeled standard compound is monitored, thereby confirming the relative binding affinity of the test compound.

[0192] kit The Disclosure also includes a pharmaceutical kit useful, for example, in the treatment or prevention of PI3K-related diseases or disorders such as cancer, which comprises one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of salts and crystalline forms of the Disclosure. As will be apparent to those skilled in the art, such a kit may further include, as necessary, one or more of a variety of conventional pharmaceutical kit components, such as additional containers having one or more pharmaceutically acceptable carriers. Instructions indicating the amount of the component to be administered, guidelines for administration, and / or guidelines for mixing the components may also be included in the kit, either as an insert or a label.

[0193] The present invention will be described in more detail using specific examples. The following examples are provided for illustrative purposes only and are not intended to limit the invention in any way. Those skilled in the art will readily recognize a variety of non-essential parameters that can be changed or modified to produce essentially the same results. The salts and crystalline forms of the examples were found to be PI3Kγ inhibitors by at least one assay described herein. [Examples]

[0194] Preparative LC-MS purification of a portion of the prepared compounds was performed using Waters mass fractionation systems. The basic equipment setup, protocols, and control software for the operation of these systems are 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 of B from 2% to 80% over 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 an MS detector or by flash chromatography (silica gel), as shown in the examples. Typical column conditions for preparative reversed-phase high-performance liquid chromatography (RP-HPLC) are as follows: Purification at pH=2: Waters Sunfire (trademark) C 18 5μm, 30×100mm or Waters XBridge® C 18 The elution parameters were 5 μm, 30 × 100 mm column, mobile phase A: 0.1% TFA (trifluoroacetic acid) in water, and mobile phase B: acetonitrile; the flow rate was 60 mL / min, and the separation gradient 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, with mobile phase A: 0.1% NH4OH 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, 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)).

[0197] In the following example, X-ray powder diffraction analysis was performed using a Bruker D8 Advance ECOX powder diffractometer (XRPD) with the following parameters: a 1.5418 Å Cu source, a LYNXEYETM detector, and an X-ray output of 40 kV, 25 mA. The 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 using a TA Instrument Discovery DSC2500 differential scanning calorimetry meter equipped with an autosampler. Typical experimental conditions were as follows: 20–300°C at 10°C / min, nitrogen gas flow at 50 mL / min, and aluminum sample pans.

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

[0200] Example 1.2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide [ka] Step 1. 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethane-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 obtain a yellowish-brown oil. Purification by flash column chromatography using methyl tert-butyl ether (MTBE) (0% to 50%) in hexane yielded the desired product (8.42 g, 95.7%) as a yellow oil. C9H7BrF3(M-OH) + LCMS for the following values: m / z = 251.0, 253.0; measured values: 250.9, 252.8.

[0201] Step 2.1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethane-1-one [ka] A mixture of 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethane-1-ol (8.41 g, 31.3 mmol) in 125 mL of dichloromethane at 0°C was treated with Dess-Martin periodinane (19.9 g, 46.9 mmol) and stirred at RT for 2.5 hours. The reaction mixture was concentrated (by rotary evaporation using a water bath set at 30°C) to obtain an oily solid, which was diluted with 200 mL of diethyl ether to precipitate more solid. This mixture was filtered through Celite®, and the Celite® was rinsed with an additional 200 mL of diethyl ether. The filtrate was washed with saturated sodium bicarbonate solution (3 × 200 mL) and brine, dried over sodium sulfate, filtered, and concentrated to obtain an oily solid. The oily solid was partitioned between 150 mL of diethyl ether and 100 mL of water. The organic layer was separated, washed with saturated sodium bicarbonate solution (2 × 75 mL) and brine, dried over sodium sulfate, filtered, and concentrated to obtain the desired product (7.93 g, 95.0%) as oil, which was used without further purification. C9H7BrF3O (M+H) + LCMS for the following values: m / z = 267.0, 269.0; measured values: 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-trifluoroethane-1-one (7.92 g, 29.7 mmol) in dichloromethane (29.7 mL) was treated with trimethylsilyl cyanide (8.70 mL, 65.2 mmol), potassium cyanide (0.290 g, 4.45 mmol), and 18-crown-6 (0.290 g, 1.10 mmol), and stirred for 1 hour. Due to the exothermic reaction after the addition of 18-crown-6, the reaction was cooled in an ice bath. The reaction mixture was concentrated (by rotary evaporation using a water bath set to 28°C) to obtain 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 hours. The reaction mixture was diluted with water (75 mL) and extracted with diethyl ether (3 × 75 mL). The combined organic extract was washed with brine, dried over sodium sulfate, filtered, and concentrated. Reconcentration from hexane yielded the desired product (8.70 g, 99.8%) as an orange solid, which was used without further purification. C9H7BrF3O (M-CN) + LCMS for the following: m / z = 267.0, 269.0; measured values: 266.9, 269.0.

[0203] Step 4. 2-(3-bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (the second enantiomer to elute) [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 bubbling through the reaction mixture for 45 minutes while stirring at 0°C. The cooling bath was removed, and the reaction mixture was stirred at room temperature for 61 hours. Nitrogen was bubbling through the reaction mixture for 10 minutes to remove some of the HCl, and the 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 the remaining solid. 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 obtain a brown oil. The racemic product was purified by flash column chromatography using MTBE (0% to 60%) in hexane to obtain a yellow oily solid. The racemic mixture was separated by preparative chiral HPLC (Phenomenex Lux Amylose-1 [21.2 × 250 mm, 5 micrometers], eluted with 95% ethanol in hexane at a flow rate of 18 mL / min, loaded with approximately 100 mg in 2 mL of ethanol) to obtain the desired second enantiomer (4.50 g, 48.8%) as a viscous yellow oil. The first enantiomer eluted had a retention time of 4.0 minutes. The second enantiomer eluted had a retention time of 5.3 minutes.

[0204] The second enantiomer to elute: 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 the following: m / z = 312.0, 314.0; measured values: 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 enantiomer to elute) in 1,4-dioxane (57.2 mL) was treated with bis(pinacolate)diborone (3.49 g, 13.7 mmol) and potassium acetate (3.71 g, 37.8 mmol), and degassed under 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 an 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 (0% to 100%) in hexane yielded 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 this value: m / z = 360.2; measured value: 360.1.

[0206] Step 6.2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide A solution of 3-bromo-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-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 under nitrogen for 5 minutes, treated with dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloromethane adduct (3.27 g, 4.00 mmol), degassed further under nitrogen for 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 with 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 extract was dried over sodium sulfate, filtered, and concentrated to a brown foam. Purification by flash column chromatography using MeOH (0% to 10%) in dichloromethane yielded the desired product as a red / brown foam, which was not completely pure. This material was re-purified by flash column chromatography using MeOH (0% to 15%) in dichloromethane to obtain the desired product as an orange / brown foam, which was still not completely pure. This material was re-purified by flash column chromatography using ethyl acetate (0% to 100%) (containing 5% MeOH) in hexane to obtain the desired product as a white foam, which also still contained impurities. This material was re-purified by flash column chromatography using acetonitrile (0% to 100%) (containing 5% MeOH) in dichloromethane to obtain 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 this value: m / z = 434.1; measured value: 434.1.

[0207] Example 2: Preparation and characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IA (free base). 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (0.050 g, 0.115 mmol) was placed in a vial, and a pre-mixed solution of isopropyl acetate (0.676 mL) / heptane (1.34 mL) in a 1:2 ratio was added dropwise while stirring at 80°C. After adding 2 mL, the solid did not completely dissolve and some remained at the bottom of the vial. After almost all of the solid had dissolved, new solid had formed on the wall of the vial. After stirring at 80°C for 2 hours, further solid formed. After cooling to ambient temperature, the solid was filtered and washed with heptane. The solid was collected and dried under reduced pressure for 30 minutes to obtain 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (form IA) (33.2 mg, 66.4%) as a white solid.

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

[0209] DSC analysis of Form IA revealed one endothermic peak with an onset temperature of 191.9°C and a maximum at 193.2°C. The DSC thermogram is shown in Figure 2.

[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 Figure 3.

[0211] Form IA was confirmed to be 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 while stirring at ambient temperature. The mixture was heated at 80°C for 2 hours with stirring. The mixture was cooled to ambient temperature and stirred for 1 hour. The solid was collected by filtration and air-dried to give 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (Form IIA).

[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 are shown in Table 2 below. Table 2

[0214] DSC analysis of morphology IIA revealed a single endothermic peak with a maximum temperature of 179.7°C at an initial temperature of 177.2°C. The DSC thermogram is shown in Figure 5.

[0215] TGA analysis of morphology IIA revealed significant weight loss at temperatures above 200°C due to sample degradation. The TGA thermogram is shown in Figure 6.

[0216] Morphology IIA was confirmed to be an anhydrous, solvated crystalline form.

[0217] Example 4: Preparation and characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, crystalline form IIIA (free base). Approximately 72 mg of the free base of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide was dissolved in 1 mL of MeOH in a 4 mL clear glass vial. The solution was evaporated at ambient temperature. The resulting solid, which is a MeOH solvate, was dried overnight at 60°C under vacuum to obtain 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (form IIIA).

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

[0219] DSC analysis of morphology IIIA revealed a single endothermic peak with a maximum temperature of 143.0°C at an initial temperature of 134.3°C. The DSC thermogram is shown in Figure 8.

[0220] TGA analysis of morphology IIIA revealed significant weight loss at temperatures above 200°C due to sample degradation. The TGA thermogram is shown in Figure 9.

[0221] Morphology IIIA was confirmed to be an anhydrous, 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] N-iodosuccinimide (0.45 g, 2.0 mmol) was added 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). The reaction mixture was then heated at 60°C for 15.5 hours. The reaction mixture was concentrated under vacuum. The resulting solid was taken into dichloromethane (DCM). The organic layer was successively washed with water and saturated Na₂S₂O₃(aq). The organic layer was then dried over Na₂SO₄, filtered, and concentrated to obtain the title compound as a pale yellow solid (0.64 g, 88%). C₆H₃Br₂IN₃(M+H) + LCMS for the following: Calculated m / z = 401.8, 403.8, 405.8; measured values ​​401.8, 403.7, 405.6.

[0223] Step 2.6-Bromo-3-iodo-N-(4-methoxybenzyl)imidazo[1,2-a]pyrazine-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 by 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. C 14 H 13 BrIN4O (M+H) + LCMS: 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

Chemical Formula

[0225] Step 4. 2-(3-bromo-4-methylphenyl)-1,1,1-trifluoropropan-2-ol

Chemical Formula

[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(pinacolate)diborone (0.294 g, 1.16 mmol), and potassium acetate (0.288 g, 2.94 mmol) in tetrahydrofuran (4.95 mL) was degassed under nitrogen for 5 minutes. The reaction mixture was treated with triphenylphosphine palladium chloride (0.025 g, 0.036 mmol), degassed further under nitrogen for 5 minutes, and heated at 135°C in a microwave for 20 minutes. The reaction mixture was diluted with ethyl acetate, filtered through a 0.5 micrometer cartridge, and rinsed with ethyl acetate. The filtrate was washed with water and brine, dried over sodium sulfate, filtered, and concentrated to obtain the crude residue. Purification by flash column chromatography using ether in hexane (0% to 50%) yielded the desired product (272 mg, 92.5%) as a colorless oil. 16 H 23 BF3O3(M+H) + LCMS for this value: m / z = 331.2; measured value: 331.2.

[0227] Step 6.2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-1,1,1-trifluoropropane-2-ol [ka] A mixture of 6-bromo-3-iodoimidazo[1,2-a]pyrazine-8-aminetrifluoroacetate (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 the ethanol and diluted with ethyl acetate and water. The solid was removed by filtration, the aqueous layer of the filtrate was separated and extracted with ethyl acetate (2×). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated to obtain the crude residue. Purification by flash column chromatography using methanol (0%~2%) in dichloromethane yielded the desired product (610 mg, 77.8%) as a white foam. 16 H 15 BrF3N4O (M+H) + LCMS for the following: m / z = 415.0, 417.0; measured values: 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]pyrazine-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)2CH2Cl2 (0.049 g, 0.060 mmol), degassed further with nitrogen for 5 minutes, saturated with CO by bubbling gas under the reaction surface for 3 minutes, and heated overnight at 60°C. 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 (2×). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated to obtain a brown oil. Purification by flash column chromatography using methanol (0%-4%) in dichloromethane yielded the desired product (158 mg, 66.5%) as a yellowish-brown oily solid. 18 H 18 F3N4O3(M+H) + LCMS for this value: m / z = 395.1; measured value: 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 overnight at 80°C. The reaction mixture was treated with an additional trimethylaluminum (0.70 mL, 1.40 mmol) (2 M in toluene) and stirred overnight at 80°C. The reaction mixture was cooled to room temperature, diluted with methanol, and filtered through a Celite® pad. After rinsing with MeOH (2×), the filtrate was concentrated to obtain a yellowish-brown oil. Purification by silica gel chromatography (0-5% MeOH / DCM) yielded 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], eluted with 12% ethanol in hexane at a flow rate of 18 mL / min, loaded with approximately 8 mg in 800 μL of ethanol). The first eluted peak had a retention time of 11.9 minutes (Example 1; Enantiomer 1). The second eluted peak 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 this value: m / z = 452.2; measured value: 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 this: m / z = 452.2; measured value: 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). 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 (Example 6, enantiomer 2 of Step 8; 184 g, 408 mmol) and isopropyl acetate (950 mL) were placed in a round-bottom flask. The mixture was stirred at 80°C for 1 hour, cooled to room temperature (RT), and stirred overnight at RT. The solid was collected to obtain 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] Morphology IB was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of morphology IB is shown in Figure 10, and the peak data is shown in Table 4 below. [Table 4]

[0234] DSC analysis of morphology IB revealed a single endothermic peak with a maximum temperature of 174.2°C at an initiation temperature of 172.2°C. The DSC thermogram is shown in Figure 11. Morphology IB was confirmed to be an anhydrous, solvated 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). 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 (Example 6, enantiomer 2 of Step 8; 252 mg, 0.559 mmol) and isopropyl acetate (1.25 mL) were placed in a vial, and the solid was slowly dissolved. The mixture was treated with heptane (0.35 mL) until the solid remained. The mixture was heated at 80°C for 30 minutes and stirred overnight at RT. The solid was recovered to obtain 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] Morphology IIB was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of morphology IIB is shown in Figure 12, and the peak data is shown in Table 5 below. [Table 5]

[0237] DSC analysis of morphology IIB revealed a single endothermic peak with a maximum temperature of 165.4°C at an initiation temperature of 161.7°C. The DSC thermogram is shown in Figure 8. Morphology IIB was confirmed to be an anhydrous, solvated 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)ethane-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 minutes, treated with iodomethane-d3 (1.44 mL, 23.2 mmol), and stirred overnight at 45°C. The reaction mixture was cooled to rt and diluted with water and ethyl acetate. The aqueous layer was separated and extracted with ethyl acetate (2×). The combined organic extracts were washed with water and brine, dried over magnesium sulfate, filtered, and concentrated (in a 60-70 Tor bath at 25°C) to obtain 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 this value: m / z = 138.1; measured value: 138.1.

[0239] Step 2. 1-(3-Bromo-4-(methyl-d3)phenyl)ethan-1-one

Chemical Structure

[0240] Step 3.2-(3-bromo-4-(methyl-d3)phenyl)-1,1,1-trifluoropropane-2-ol [ka] A solution of 1-(3-bromo-4-(methyl-d3)phenyl)ethane-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 obtain the crude residue. Purification by flash column chromatography using ethyl acetate in hexane (0%-20%) yielded 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(pinacolate)diborone (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 under nitrogen for 10 minutes. The reaction mixture was treated with bis(triphenylphosphine)palladium(II) dichloride (1.16 g, 1.67 mmol), degassed further under nitrogen for 10 minutes, and stirred overnight at 100°C. 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 again with nitrogen for 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 obtain a brown oil. Purification by flash column chromatography using MTBE (0%~20%) in hexane yielded 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 this value: m / z = 334.2; measured value: 334.3.

[0242] Step 5.2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazine-3-yl)-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol (racemic mixture) Chem. 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 (107 mL, 107 mmol) in water, 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 give a darkened oil. Purification by flash column chromatography using methanol (0% to 5%) in dichloromethane followed by repurification by flash column chromatography using ethyl acetate (0% to 100%) in hexane afforded 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 Chem. A racemic mixture of 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazine-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], eluted at a flow rate of 20 mL / min in 20% ethanol in hexane, loaded with approximately 200 mg in 4 mL of ethanol). The first eluted peak had a retention time of 9.6 minutes. The second eluted peak had a retention time of 14.6 minutes.

[0244] Peak 2: 1 ¹H 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 C16H12D3BrF3N4O (M+H)+: m / z = 418.1, 420.1; measured values: 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 (prepared single enantiomer) [ka] A solution of 2-(3-(8-amino-6-bromoimidazo[1,2-a]pyrazine-3-yl)-4-(methyl-d3)phenyl)-1,1,1-trifluoropropan-2-ol (peak 2 in 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 minutes. The reaction mixture was treated with Pd(dppf)2CH2Cl2 (0.998 g, 1.22 mmol), degassed further with nitrogen for 5 minutes, saturated with CO by bubbling gas under the reaction surface for 3 minutes, and heated overnight at 60°C. 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 (3×). The combined organic layers were washed with water, saturated ammonium chloride solution, and brine, dried over magnesium sulfate, filtered, and concentrated to obtain a brown oil. Purification by flash column chromatography using methanol (0%-5%) in dichloromethane yielded the desired product (4.49 g, 92.4%) as an orange solid. 18 H 15 D3F3N4O3(M+H) + LCMS for this value: m / z = 398.1; measured value: 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 (prepared single enantiomer) [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 layer was extracted with additional 1.0 M sodium hydroxide (3 × 20 mL). The combined basic aqueous layer was adjusted to pH approximately 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 obtain the desired product (4.06 g, 93.8%) as a yellowish-brown 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 this value: m / z = 384.1; measured value: 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 minutes, treated with triethylamine (4.43 mL, 31.8 mmol), and stirred at room temperature for 3.5 hours. The reaction mixture was diluted with water (500 mL) and brine (100 mL) and extracted with ethyl acetate (3 × 150 mL). The combined organic matter was washed with saturated ammonium chloride (150 mL), 11% sodium carbonate (150 mL), and brine (100 mL), dried over magnesium sulfate, filtered, and concentrated to obtain a yellowish-brown oil. The desired product (4.28 g, 89.0%) was obtained as foam by purification using flash column chromatography with methanol (0%~5%) in dichloromethane. 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; measured value: 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). 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) were placed in a round-bottomed container and heated to 80°C. The mixture was stirred at 80°C and solid formation began within 5 minutes. The mixture was stirred at 80°C for 1 hour. Heating was stopped and the mixture was stirred for 1 hour while simultaneously cooling to rt. The mixture was treated with heptane (25.5 mL) droplets from a dropper funnel for 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 obtain 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] The morphological IC was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of the morphological IC is shown in Figure 14, and the peak data is shown in Table 6 below. [Table 6]

[0250] DSC analysis of the IC morphology revealed a single endothermic peak with a maximum temperature of 179.0°C at an initiation temperature of 173.4°C. The DSC thermogram is shown in Figure 15. The IC morphology was confirmed to be anhydrous, solvated crystalline form.

[0251] Example 11. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide (HBr) salt 98.81 mg of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide free base 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 thoroughly mixed. The solution was evaporated at room temperature to obtain HBr salt crystals.

[0252] Example 12. Characterization of single crystal 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-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 × 0.210 × 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) = 1.98 mm -1 .

[0253] Data Acquisition: Data acquisition was performed using a Bruker SMART APEX-II CCD system, MoK alpha beam, standard focus tube, anode output = 50kV × 30mA, crystal-to-plate distance = 5.0cm, 512 × 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 minimum / maximum = (-26, 26, -12, 13, -27, 27), data input to shelx = 38968, unique data = 9756, 2-theta range = 4.51 to 55.43°, completeness for 2-theta 55.43° = 99.60%, R(int-xl) = 0.0672, and SADABS correction applied.

[0254] Analysis and refinement: The crystal structure was analyzed using XS (Shelxtl) and refined using the shelxtl software package. Refinement was performed using F 2 Complete matrix least squares method for the following, scattering coefficients from Tables 4.2.6.8 and 6.1.1.4 of Int. Tab. Vol C, number of data points = 9756, limit number = 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 peak and Hall = 1.795 and -0.642 e / Å 3 The refinement was performed using a flack parameter of 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 ×10 3 Table 8 shows the bond distance [Å] and angle [degrees]. Table 9 shows the anisotropy displacement parameter [Å]. 2 ×10 3 ) indicates.

[0255] Results: This analysis confirmed the structure of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide. The asymmetric unit, as shown in Figures 16A-16B, contains two molecules of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, two bromides to maintain charge balance, and one methanol solvent molecule. Enantiomerization was based on the Flack parameter refined to 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 measured in the presence or absence of a certain concentration range of the test compound in RPMI (2 × 10⁶ cells per 90 μL). 5Plate the cells (per well) into a 96-well flat-bottom tissue culture plate (Corning, Corning, NY). Cover the plate and incubate at 37°C and 5% CO2 for 2 hours, then treat with or without 10 nM MCP-1 (MYBioSource, San Diego, CA) at 37°C and 5% CO2 for 15 minutes. Centrifuge the plate at 1600 RPM and remove the supernatant. Lyse the cells on wet ice for 30 minutes in a lysis buffer (Cell Signaling, Danvers, MA) containing a protease inhibitor (Calbiochem / EMD, Germany), PMSF (Sigma, St Louis MO), and HALTS (Thermo Fisher, Rockford, IL). Freeze the cell lysates at -80°C before testing. Test the lysates with human / mouse / rat Phospho-RPS6 ELISA (R&D Systems, Inc. Minn, MN). The plates are measured using a microplate reader (SpectraMax M5-Molecular Devices, LLC Sunnyvale, CA) with a wavelength correction of 540 set to 450 nm. 50 This determination is made using GraphPad Prism 5.0 software by fitting the inhibitor inhibition percentage curve to the logarithm of the inhibitor concentration.

[0257] Example B. Scintillation proximity assay of PI3K-γ material [γ- 33P]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-phosphorus bond (PIP2), CAS 204858-53-7, was purchased from Echelon Biosciences (Salt Lake City, UT). Recombinant human protein PI3Kγ (p110γ) was purchased from Life Technology (Grand Island, NY). ATP, MgCl2, DTT, EDTA, MOPS, and CHAPS were purchased from SigmaAldrich (St. Louis, MO).

[0258] The kinase reaction was performed in a 384-well polystyrene Greiner Bio-one white plate from Thermo Fisher Scientific, with a final volume of 25 μL. The inhibitor was first serially diluted in DMSO and added to the plate wells, after which other reactants were added. The final concentration of DMSO in the assay was 2%. The PI3Kγ assay was performed at room temperature in 20 mM MOPS (pH 6.7), 10 mM MgCl2, 5 mM DTT, and 0.03% CHAPS. The reaction was initiated by the addition of ATP, and the final reaction mixture consisted of 20 μM PIP2, 2 μM ATP, and 0.5 μCi [γ- 33 The reaction consisted of [P]ATP and 13 nM PI3Kγ. The reaction mixture was incubated for 120 minutes and terminated by adding 40 μL of SPA beads suspended in a reaction termination 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 plate, it was shaken overnight at room temperature and centrifuged at 1500 rpm for 10 minutes. The radioactivity of the product was determined by scintillation counting using a Topcount (Perkin-Elmer). IC 50The determination was made using GraphPad Prism 6.0 software by fitting the percentage activity curve of the solvent control to the logarithm of the inhibitor concentration.

[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-phosphorus bond (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 SigmaAldrich (St. Louis, MO).

[0260] The kinase reaction was performed in a 384-well polystyrene Greiner Bio-one white plate from Thermo Fisher Scientific, with a final volume of 25 μL. The inhibitor was first serially diluted in DMSO and added to the plate wells, after which other reactants were added. The final concentration of DMSO in the assay was 2%. The PI3Kδ assay was performed at room temperature in 20 mM MOPS (pH 6.7), 10 mM MgCl2, 5 mM DTT, and 0.03% CHAPS. The reaction was initiated by the addition of ATP, and the final reaction mixture consisted of 20 μM PIP2, 2 μM ATP, and 0.5 μCi [γ- 33The reaction consisted of [P]ATP and 3.4 nM PI3Kδ. The reaction mixture was incubated for 120 minutes and terminated by adding 40 μL of SPA beads suspended in a reaction termination 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 plate, it was shaken overnight at room temperature and centrifuged at 1500 rpm for 10 minutes. The radioactivity of the product was determined by scintillation counting using a Topcount (PerkinElmer). IC 50 The determination was made using GraphPad Prism 6.0 software by fitting the percentage activity curve of the solvent control to the logarithm of the inhibitor concentration.

[0261] The compounds of Examples 1, 5, 6, and 9 were tested in the assays described in Examples A, B, and C, and the ICs shown in Table A below are obtained. 50 It was found that it has a value. [Table 10] + indicates ICs with a minimum impedance of ≤100nM 50 This refers to ICs with a minimum impedance of ≤500nM. 50 This refers to an IC with a power output of <2000nM. 50 This refers to an IC with a minimum impedance of ≥2000nM. 50 It refers to. # is an IC with a minimum impedance of ≤100nM. 50 This refers to ICs with a minimum impedance of ≤500nM. 50 This refers to ICs with a minimum impedance of <1000nM. 50 This refers to an IC with a minimum impedance of ≥1000nM. 50 It refers to.

[0262] In addition to those described herein, various modifications of the present invention will be 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. All references, including all patents, patent applications, and publications cited herein, are incorporated herein by reference in their entirety. Furthermore, this application also encompasses the following aspects. [Aspect 1] Crystalline form of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide. [Aspect 2] The crystalline form according to Embodiment 1, which is anhydrous and solvated. [Aspect 3] The crystal form according to embodiment 1 or 2, which is form IA. [Aspect 4] A crystal morphology according to any one of embodiments 1 to 3, having an X-ray powder diffraction pattern including at least one peak at 8.6°±0.2° in 2θ. [Aspect 5] The crystal morphology according to any one of embodiments 1 to 4, having an X-ray powder diffraction pattern including at least one peak at 9.5°±0.2° in 2θ. [Aspect 6] The crystal morphology has an X-ray powder diffraction pattern at 2θ that includes the following peaks: 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°, as described in any one of embodiments 1 to 5. [Aspect 7] The crystal morphology described in any one of embodiments 1 to 6, having an X-ray powder diffraction pattern with four or more 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°; 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°. [Aspect 8] A crystalline form according to any one of embodiments 1 to 7, having an X-ray powder diffraction pattern substantially as shown in Figure 1. [Aspect 9] A crystal morphology according to any one of embodiments 1 to 8, having a DSC thermogram that includes an endothermic peak having a maximum value at approximately 193°C. [Aspect 10] The crystal morphology according to any one of embodiments 1 to 9, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 2. [Aspect 11] The crystalline form according to any one of embodiments 1 to 10, having a thermogravimetric analysis (TGA) substantially as shown in Figure 3. [Aspect 12] The crystal form according to embodiment 1 or 2, which is form IIA. [Aspect 13] A crystal morphology according to any one of embodiments 1, 2, and 12, having an X-ray powder diffraction pattern including at least one peak at 9.1°±0.2° in 2θ. [Aspect 14] The crystal morphology according to any one of embodiments 1, 2, 12, and 13, having an X-ray powder diffraction pattern including at least one peak at 11.1° ± 0.2° at 2θ. [Aspect 15] A crystal morphology according to any one of embodiments 1, 2, and 12 to 14, having an X-ray powder diffraction pattern including at least one peak at 12.6° ± 0.2° at 2θ. [Aspect 16] A crystal morphology according to any one of embodiments 1, 2, and 12 to 15, having an X-ray powder diffraction pattern including at least one peak at 13.5° ± 0.2° at 2θ. [Aspect 17] The crystal morphology described in any one of embodiments 1, 2, and 12 to 16 has an X-ray powder diffraction pattern containing the following peaks at 2θ: 9.1°±0.2°; 11.1°±0.2°; 12.6°±0.2°; and 13.5°±0.2°. [Aspect 18] The crystal morphology has an X-ray powder diffraction pattern in which four or more peaks at 2θ are included: 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°, as described in any one of embodiments 1, 2, and 12 to 17. [Aspect 19] A crystalline form according to any one of embodiments 1, 2, and 12 to 18, having an X-ray powder diffraction pattern substantially as shown in Figure 4. [Aspect 20] A crystalline form according to any one of embodiments 1, 2, and 12 to 19, having a DSC thermogram that includes an endothermic peak having a maximum at approximately 180°C. [Aspect 21] A crystalline morphology according to any one of embodiments 1, 2, and 12 to 20, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 5. [Aspect 22] A crystalline morphology according to any one of embodiments 1, 2, and 12 to 21, having a thermogravimetric analysis (TGA) substantially as shown in Figure 6. [Aspect 23] The crystal form according to embodiment 1 or 2, which is form IIIA. [Aspect 24] The crystal morphology according to any one of embodiments 1, 2, and 23, having an X-ray powder diffraction pattern including at least one peak at 8.1°±0.2° in 2θ. [Pattern 25] A crystal morphology according to any one of embodiments 1, 2, 23, and 24, having an X-ray powder diffraction pattern including at least one peak at 10.6° ± 0.2° in 2θ. [Aspect 26] A crystal morphology according to any one of embodiments 1, 2, and 23 to 25, having an X-ray powder diffraction pattern including at least one peak at 13.5° ± 0.2° at 2θ. [Aspect 27] The crystal morphology described in any one of embodiments 1, 2, and 23 to 26 has an X-ray powder diffraction pattern containing the following peaks at 2θ: 8.1°±0.2°; 10.6°±0.2°; 13.5°±0.2°; and 14.2°±0.2°. [Aspect 28] The crystal morphology has an X-ray powder diffraction pattern in which four or more peaks at 2θ are included: 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°, as described in any one of embodiments 1, 2, and 23 to 27. [Aspect 29] A crystalline morphology according to any one of embodiments 1, 2, and 23 to 28, having an X-ray powder diffraction pattern substantially as shown in Figure 7. [Aspect 30] A crystalline form according to any one of embodiments 1, 2, and 23 to 29, having a DSC thermogram that includes an endothermic peak having a maximum at approximately 143°C. [Aspect 31] A crystalline morphology according to any one of embodiments 1, 2, and 23 to 30, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 8. [Aspect 32] A crystalline morphology according to any one of embodiments 1, 2, and 23 to 31, having a thermogravimetric analysis (TGA) substantially as shown in Figure 9. [Aspect 33] The 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. [Aspect 34] The crystalline form according to embodiment 33, which is anhydrous and solvated. [Aspect 35] The crystal form according to embodiment 33 or 34, which is form IB. [Aspect 36] The crystal morphology according to any one of embodiments 33 to 35, having an X-ray powder diffraction pattern including at least one peak at 6.2°±0.2° in 2θ. [Aspect 37] The crystal morphology according to any one of embodiments 33 to 36, having an X-ray powder diffraction pattern including at least one peak at 15.6°±0.2° in 2θ. [Aspect 38] The crystal morphology has an X-ray powder diffraction pattern at 2θ that includes the following peaks: 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°, as described in any one of embodiments 33 to 37. [Aspect 39] The crystal morphology described in any one of embodiments 33 to 37, having an X-ray powder diffraction pattern with four or more 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°; 16.0°±0.2°; 16.7°±0.2°; 20.7°±0.2°; and 23.2°±0.2°. [Aspect 40] A crystalline morphology according to any one of embodiments 33 to 39, having an X-ray powder diffraction pattern substantially as shown in Figure 10. [Aspect 41] The crystal morphology according to any one of embodiments 33 to 40, having a DSC thermogram that includes an endothermic peak having a maximum at approximately 174°C. [Aspect 42] The crystal morphology according to any one of embodiments 33 to 41, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 11. [Aspect 43] The crystal form described in embodiment 33 or 34, which is form IIB. [Aspect 44] The crystal morphology according to any one of embodiments 33, 34, and 43, having an X-ray powder diffraction pattern including at least one peak at 4.2°±0.2° in 2θ. [Aspect 45] A crystal morphology according to any one of embodiments 33, 34, 43, and 44, having an X-ray powder diffraction pattern including at least one peak at 13.3°±0.2° in 2θ. [Aspect 46] A crystal morphology according to any one of embodiments 33, 34, and 43 to 45, having an X-ray powder diffraction pattern including at least one peak at 17.0° ± 0.2° at 2θ. [Aspect 47] A crystal morphology according to any one of embodiments 33, 34, and 43 to 46, having an X-ray powder diffraction pattern including at least one peak at 18.8° ± 0.2° at 2θ. [Aspect 48] The crystal morphology described in any one of embodiments 33, 34, and 43 to 47 has an X-ray powder diffraction pattern containing the following peaks at 2θ: 4.3°±0.2°; 7.4°±0.2°; 13.3°±0.2°; and 15.3°±0.2°. [Aspect 49] The crystal morphology has an X-ray powder diffraction pattern in which four or more of the following peaks are included at 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°, as described in any one of embodiments 33, 34, and 43 to 47. [Aspect 50] A crystalline morphology according to any one of embodiments 33, 34, and 43 to 49, having an X-ray powder diffraction pattern substantially as shown in Figure 12. [Aspect 51] A crystalline form according to any one of embodiments 33, 34, and 43 to 50, having a DSC thermogram that includes an endothermic peak having a maximum at approximately 165°C. [Aspect 52] A crystalline morphology according to any one of embodiments 33, 34, and 43 to 50, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 13. [Aspect 53] Compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 The crystalline form of )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide. [Aspect 54] The crystalline form according to embodiment 53, which is anhydrous and solvated. [Aspect 55] The crystal form according to embodiment 53 or 54, which is an IC form. [Aspect 56] The crystal morphology according to any one of embodiments 53 to 55, having an X-ray powder diffraction pattern including at least one peak at 6.2°±0.2° in 2θ. [Aspect 57] The crystal morphology according to any one of embodiments 53 to 56, having an X-ray powder diffraction pattern including at least one peak at 11.9°±0.2° in 2θ. [Aspect 58] The crystal morphology has an X-ray powder diffraction pattern at 2θ that includes the following peaks: 6.2°±0.2°; 10.4°±0.2°; 11.3°±0.2°; 11.9°±0.2°; and 12.5°±0.2°, as described in any one of embodiments 53 to 57. [Aspect 59] The crystal morphology has an X-ray powder diffraction pattern at 2θ that includes four or more of the following peaks: 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°, as described in any one of embodiments 53 to 58. [Aspect 60] A crystalline morphology according to any one of embodiments 53 to 59, having an X-ray powder diffraction pattern substantially as shown in Figure 14. [Aspect 61] A crystal morphology according to any one of embodiments 53 to 60, having a DSC thermogram that includes an endothermic peak having a maximum at approximately 179°C. [Aspect 62] The crystal morphology according to any one of embodiments 53 to 61, having a differential scanning calorimetry thermogram (DSC) substantially as shown in Figure 15. [Aspect 63] The salt is 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide. [Aspect 64] The salt according to embodiment 63, wherein the salt is 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide in a 1:1 stoichiometric ratio with respect to hydrobromic acid. [Aspect 65] A salt according to embodiment 63 or 64, which is substantially isolated. [Aspect 66] The salt according to embodiment 63 or 64, which is in a crystalline form. [Aspect 67] The crystal form described in embodiment 66, which is a solvated crystal form. [Pattern 68] The crystalline form according to embodiment 67, which is a methanol solvate crystalline form. [Aspect 69] A crystalline form according to any one of embodiments 1 to 62, 67, and 68, which is substantially isolated. [Aspect 70] A composition comprising the crystalline form described in any one of embodiments 1 to 62 and 67 to 69 or the salt described in any one of embodiments 63 to 66. [Aspect 71] The composition according to embodiment 70, further comprising at least one pharmaceutically acceptable carrier. [Aspect 72] A method for inhibiting the activity of PI3Kγ kinase, comprising contacting the kinase with a crystalline form described in any one of embodiments 1 to 62 and 67 to 69 or a salt described in any one of embodiments 63 to 66. [Aspect 73] The method according to embodiment 72, wherein the crystalline form is a selective inhibitor of PI3Kγ more than one or more of PI3Kα, PI3Kβ, and PI3Kδ. [Aspect 74] A method for treating a disease or disorder associated with abnormal expression or activity of PI3Kγ kinase in a patient, comprising administering to the patient a therapeutically effective amount of the crystalline form described in any one of embodiments 1 to 62 and 67 to 69 or the salt described in any one of embodiments 63 to 66. [Aspect 75] The method according to embodiment 74, wherein the disease or disorder is an autoimmune disease or disorder, cancer, cardiovascular disease, or neurodegenerative disease. [Aspect 76] The method according to aspect 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, kidney cancer, gastric cancer, seminomas, teratomas, astrocytoma, neuroblastoma, glioma, or sarcoma. [Aspect 77] The method according to embodiment 76, wherein the sarcoma is Askin tumor, staphyloid sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant Schwann cell tumor, osteosarcoma, hydatidiform soft tissue sarcoma, angiosarcoma, phyllodes cystic sarcoma, dermatofibrosarcoma protuberans, desmoid tumor, fibroplastic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), perivascular cell tumor, angiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma, or undifferentiated pleomorphic sarcoma. [Aspect 78] The method according to embodiment 74, wherein the disease or disorder is acute myeloid leukemia, acute monocytic leukemia, small lymphocytic lymphoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma, T-cell acute lymphoblastic leukemia (T-ALL), cutaneous T-cell lymphoma, large granular lymphocytic leukemia, mature (peripheral) T-cell neoplasm (PTCL), anaplastic large cell lymphoma (ALCL), or lymphoblastic lymphoma. [Aspect 79] The method according to embodiment 78, wherein the mature (peripheral) T-cell neoplasm (PTCL) is a pre-lymphocytic T-cell leukemia, a granular lymphocytic T-cell leukemia, an invasive NK-cell leukemia, mycosis fungoides / Sézary syndrome, an anaplastic large cell lymphoma (T-cell type), enteropathy-type T-cell lymphoma, adult T-cell leukemia / lymphoma, or angioimmunoblastic T-cell lymphoma. [Aspect 80] The method according to embodiment 78, wherein the anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL. [Aspect 81] The aforementioned diseases or disorders include Burkitt lymphoma, acute myeloblastic leukemia, chronic myeloid leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanocyte, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenström macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, and myelofibrosis. The method according to embodiment 74, wherein the lymphoma is mucosal-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary exudative lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammaglobulinemia of unknown significance (MGUS), or diffuse large B-cell lymphoma. [Aspect 82] The method according to embodiment 81, wherein the non-Hodgkin lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, low-grade NHL (iNHL), or aggressive NHL (aNHL). [Aspect 83] The method according to embodiment 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. [Aspect 84] The method according to embodiment 81, wherein the Burkitt lymphoma is endemic Burkitt lymphoma, sporadic Burkitt lymphoma, or Burkitt-like lymphoma. [Aspect 85] The method according to embodiment 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, Sjögren's syndrome, osteoarthritis, restenosis, or atherosclerosis. [Aspect 86] The method according to embodiment 84, wherein the disease or disorder is cardiac hypertrophy, cardiomyocyte dysfunction, acute coronary syndrome, chronic obstructive pulmonary disease (COPD), chronic bronchitis, hypertension, 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 xenotransplantation, glomerulosclerosis, progressive renal fibrosis, idiopathic thrombocytopenic purpura (ITP), autoimmune hemolytic anemia, vasculitis, systemic lupus erythematosus, lupus nephritis, pemphigus, or membranous nephropathy. [Aspect 87] The method according to embodiment 86, wherein the idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP. [Pattern 88] The method according to embodiment 86, wherein 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 cryoglobulinemia vasculitis (essential or hepatitis C virus (HCV) induced), Henoch-Schönlein purpura (HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener's granulomatosis, or anti-neutrophil cytoplasmic antibody-associated (ANCA) systemic vasculitis (AASV). [Aspect 89] The method according to embodiment 86, wherein the disease or disorder is Alzheimer's disease, central nervous system injury, or stroke. [Aspect 90] A process for preparing a crystalline form of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-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. [Aspect 91] The process according to embodiment 90, wherein the solvent comprises isopropyl acetate. [Aspect 92] The process according to embodiment 91, wherein the solvent further comprises heptane. [Aspect 93] The process according to any one of embodiments 90 to 92, further comprising heating the mixture to a temperature of about 70°C to about 90°C. [Aspect 94] The process according to embodiment 93, further comprising cooling the mixture to room temperature. [Aspect 95] The process according to embodiment 90, wherein the solvent comprises methanol. [Aspect 96] The process according to embodiment 90, further comprising heating the mixture to a temperature of about 50°C to about 70°C. [Aspect 97] The process according to embodiment 96, further comprising cooling the mixture to room temperature. [Aspect 98] A crystalline form of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, prepared according to the process described in any one of embodiments 90 to 97. [Aspect 99] The crystalline form according to embodiment 98, which is anhydrous and solvated. [Aspect 100] The crystal form according to embodiment 98 or 99, which is form IA. [Aspect 101] The crystal form according to embodiment 98 or 99, which is form IIA. [Aspect 102] The crystal form according to embodiment 98 or 99, which is form IIIA. [Aspect 103] A process for preparing the 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, the process comprising: dissolving the compound in a solvent to form a mixture; and crystallizing the compound from the mixture. [Aspect 104] The process according to embodiment 103, further comprising heating the mixture to a temperature of about 70°C to about 90°C. [Aspect 105] The process according to embodiment 103, further comprising cooling the mixture to room temperature. [Aspect 106] The process according to any one of embodiments 103 to 105, wherein the solvent comprises isopropyl acetate. [Aspect 107] The process according to embodiment 106, wherein the solvent further comprises heptane. [Aspect 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 described in any one of embodiments 103 to 107. [Aspect 109] The crystalline form according to embodiment 108, which is anhydrous and solvated. [Aspect 110] The crystal form according to embodiment 108 or 109, which is form IB. [Aspect 111] The crystal form according to embodiment 108 or 109, which is form IIB. [Aspect 112] Compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d 3 A process for preparing a crystalline form of )-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. [Aspect 113] The process according to embodiment 112, further comprising heating the mixture to a temperature of about 70°C to about 90°C. [Aspect 114] The process according to embodiment 112, further comprising cooling the mixture to room temperature. [Aspect 115] The process according to any one of embodiments 112 to 114, wherein the solvent comprises isopropyl acetate. [Aspect 116] The process according to embodiment 115, wherein the solvent further comprises heptane. [Aspect 117] Compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d) prepared according to the process described in any one of embodiments 112 to 116 3 The crystalline form of )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide. [Aspect 118] The crystalline form according to embodiment 117, which is anhydrous and solvated. [Aspect 119] The crystal form according to embodiment 117 or 118, which is an IC form. [Aspect 120] A process for preparing a hydrobromide salt of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, the process comprising: dissolving the compound in a solvent to form a mixture; and adding hydrobromic acid to the mixture. [Aspect 121] The process according to embodiment 120, wherein the solvent comprises methanol. [Aspect 122] The process according to embodiment 120 or 121, wherein the hydrobromic acid is added to the mixture as an aqueous solution of hydrobromic acid. [Aspect 123] The process according to any one of embodiments 120 to 122, wherein an excess amount of hydrobromic acid is added to the mixture based on 1 equivalent of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide. [Aspect 124] The process according to any one of embodiments 120 to 123, wherein about 1.1 to about 1.5 equivalents of hydrobromic acid are added to the mixture based on 1 equivalent of the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide. [Aspect 125] The process according to any one of embodiments 120 to 124, further comprising substantially isolating the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide. [Aspect 126] The process according to embodiment 125, wherein the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide is isolated in crystalline form. [Aspect 127] The process according to embodiment 125 or 126, wherein the 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide hydrobromide is isolated in methanol solvate crystalline form. [Aspect 128] Hydrobromide of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazine-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide, prepared according to the process described in any one of embodiments 120 to 127. [Aspect 129] A crystalline hydrobromide salt according to embodiment 128. [Aspect 130] The hydrobromide salt according to embodiment 129, which is in a solvated crystalline form. [Aspect 131] The hydrobromide salt according to embodiment 129 or 130, which is in methanol solvate crystalline form.

Claims

1. The crystalline 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 is, Morphology IB has an X-ray powder diffraction pattern in which four or more peaks are included at 2θ from among the peaks at 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°. Morphology IIB has an X-ray powder diffraction pattern that includes four or more peaks at 2θ, including peaks at 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°. It is a crystalline form selected from among The aforementioned crystal.

2. The crystal according to claim 1, which is anhydrous and solvated.

3. The crystal according to claim 1 or 2, which is of form IB.

4. The crystal according to any one of claims 1 to 3, having an X-ray powder diffraction pattern as shown in Figure 10.

5. The crystal according to any one of claims 1 to 4, having a DSC thermogram that includes an endothermic peak having a maximum at 174°C.

6. The crystal according to any one of claims 1 to 5, having a differential scanning calorimetry thermogram (DSC) as shown in Figure 11.

7. The crystal according to claim 1 or 2, which is of form IIB.

8. The crystal according to any one of claims 1, 2, and 7, having an X-ray powder diffraction pattern as shown in Figure 12.

9. The crystal according to any one of claims 1, 2, 7, and 8, having a DSC thermogram that includes an endothermic peak having a maximum at 165°C.

10. The crystal according to any one of claims 1, 2, and 7 to 9, having a differential scanning calorimetry thermogram (DSC) as shown in Figure 13.

11. Compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ３ A crystal of )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, This is a crystalline form IC having an X-ray powder diffraction pattern that includes four or more peaks among the following peaks at 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°. The aforementioned crystal.

12. The crystal according to claim 11, which is anhydrous and solvated.

13. The crystal according to claim 11 or 12, having an X-ray powder diffraction pattern as shown in Figure 14.

14. The crystal according to any one of claims 11 to 13, having a DSC thermogram that includes an endothermic peak having a maximum at 179°C.

15. The crystal according to any one of claims 11 to 14, having a differential scanning calorimetry thermogram (DSC) as shown in Figure 15.

16. An isolated crystal according to any one of claims 1 to 15.

17. A composition comprising the crystal described in any one of claims 1 to 16.

18. The composition according to claim 17, further comprising at least one pharmaceutically acceptable carrier.

19. A pharmaceutical agent for inhibiting the activity of PI3Kγ kinase, comprising the crystal described in any one of claims 1 to 16.

20. The pharmaceutical product according to claim 19, wherein the crystal is a selective inhibitor of PI3Kγ more than one or more of PI3Kα, PI3Kβ, and PI3Kδ.

21. A composition for treating a disease or disorder associated with abnormal expression or activity of PI3Kγ kinase in a patient, the composition comprising the crystal described in any one of claims 1 to 16.

22. The composition according to claim 21, wherein the disease or disorder is an autoimmune disease or disorder, cancer, cardiovascular disease, or neurodegenerative disease.

23. The composition according to claim 21, 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, kidney cancer, gastric cancer, seminomas, teratomas, astrocytoma, neuroblastoma, glioma, or sarcoma.

24. The composition according to claim 23, wherein the sarcoma is Askin tumor, staphyloid sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant Schwann cell tumor, osteosarcoma, hydatidiform soft tissue sarcoma, angiosarcoma, phyllodes cystic sarcoma, dermatofibrosarcoma protuberans, desmoid tumor, fibroplastic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), perivascular cell tumor, angiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma, or undifferentiated pleomorphic sarcoma.

25. The composition according to claim 21, wherein the disease or disorder is acute myeloid leukemia, acute monocytic leukemia, small lymphocytic lymphoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma, T-cell acute lymphoblastic leukemia (T-ALL), cutaneous T-cell lymphoma, large granular lymphocytic leukemia, mature (peripheral) T-cell neoplasm (PTCL), anaplastic large cell lymphoma (ALCL), or lymphoblastic lymphoma.

26. 26. The composition of claim 25, 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.

27. The composition according to claim 25, wherein the anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL.

28. The aforementioned diseases or disorders include Burkitt lymphoma, acute myeloblastic leukemia, chronic myeloid leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanocyte, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenström macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, The composition according to claim 21, wherein the lymphoma is a mucosal-associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphomatoid granulomatosis, splenic marginal zone lymphoma, primary exudative lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammaglobulinemia of unknown significance (MGUS), or diffuse large B-cell lymphoma.

29. The composition according to claim 28, wherein the non-Hodgkin lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, low-grade NHL (iNHL), or aggressive NHL (aNHL).

30. The composition according to claim 28, 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.

31. The composition according to claim 28, wherein the Burkitt lymphoma is endemic Burkitt lymphoma, sporadic Burkitt lymphoma, or Burkitt-like lymphoma.

32. The composition according to claim 21, 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, Sjögren's syndrome, osteoarthritis, restenosis, or atherosclerosis.

33. The composition according to claim 21, wherein the disease or disorder is cardiac hypertrophy, cardiomyocyte dysfunction, acute coronary syndrome, chronic obstructive pulmonary disease (COPD), chronic bronchitis, hypertension, 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 xenotransplantation, glomerulosclerosis, progressive renal fibrosis, idiopathic thrombocytopenic purpura (ITP), autoimmune hemolytic anemia, vasculitis, systemic lupus erythematosus, lupus nephritis, pemphigus, or membranous nephropathy.

34. The composition according to claim 33, wherein the idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP.

35. The composition according to claim 33, wherein 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 cryoglobulinemia vasculitis (essential or hepatitis C virus (HCV) induced), Henoch-Schönlein purpura (HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener's granulomatosis, or anti-neutrophil cytoplasmic antibody-associated (ANCA) systemic vasculitis (AASV).

36. The composition according to claim 21, wherein the disease or disorder is Alzheimer's disease, central nervous system injury, or stroke.

37. A process for preparing crystals 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, the process comprising: dissolving the compound in a solvent to form a mixture; and crystallizing the compound from the mixture.

38. The process according to claim 37, further comprising heating the mixture to a temperature of 70°C to 90°C.

39. The process according to claim 37, further comprising cooling the mixture to room temperature.

40. The process according to any one of claims 37 to 39, wherein the solvent comprises isopropyl acetate.

41. The process according to claim 40, wherein the solvent further comprises heptane.

42. Compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ３ A process for preparing crystals of )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, the process comprising: dissolving the compound in a solvent to form a mixture; and crystallizing the compound from the mixture.

43. The process according to claim 42, further comprising heating the mixture to a temperature of 70°C to 90°C.

44. The process according to claim 42, further comprising cooling the mixture to room temperature.

45. The process according to any one of claims 42 to 44, wherein the solvent comprises isopropyl acetate.

46. The process according to claim 45, wherein the solvent further comprises heptane.

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