EA044976B1 - CRYSTAL FORMS OF PHOSPHOINOSITIDE 3-KINASE (PI3K) INHIBITOR - Google Patents

Description

Field of technology to which the invention relates

The invention relates to salts and crystalline forms of 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2 -hydroxypropanamide, 8amino-N-(2-hydroxy-2-methylpropyl)-3 -(2-methyl-5 -(1,1,1 -trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2 -a]pyrazine-6-carboxamide and 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ₃ )-5-(1,1,1trifluoro-2-hydroxypropane- 2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, which are PI3K inhibitors used in the treatment of malignancies and other diseases.

State of the art

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

are phosphoinositide 3-kinase (PI3K) inhibitors used in the treatment of various diseases, including malignancies. It is usually an advantage in drug development to use a form of the drug that has the desired properties with respect to preparation, purification, reproducibility, stability, bioavailability and other characteristics. Accordingly, the salts and crystalline forms of the compounds of Formula A, Formula B and Formula C provided herein address the current need for the development of PI3K inhibitors for the treatment of serious diseases.

The essence of the invention

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

The present invention further provides a composition containing salts and crystalline forms of the compound of formula A provided herein and at least one pharmaceutically acceptable carrier. The present invention further provides a composition comprising a crystalline form of a compound of formula B and at least one pharmaceutically acceptable carrier. The present invention further provides a composition comprising a crystalline form of a compound of formula C and at least one pharmaceutically acceptable carrier.

The present invention further provides a process for preparing the salts and crystalline forms of the invention.

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

Brief description of graphic materials

In fig. 1 shows the X-ray powder diffraction (XRPD) pattern of crystalline form IA 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]nirazin-3-yl)-4-methylphenyl)-3,3 ,3-trifluoro-2-hydroxypropanamide.

In fig. Figure 2 shows the results of a differential scanning calorimetry (DSC) experiment for crystalline form IA 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4methylphenyl)-3,3 ,3-trifluoro-2-hydroxypropanamide.

In fig. Figure 3 shows the results of a thermogravimetric analysis (TGA) experiment for crystalline form IA 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]nirasin-3-yl)-4-methylphenyl)-3. 3,3trifluoro-2-hydroxypropanamide.

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In fig. Figure 4 shows the XRPD pattern of crystalline form IIA 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2 -hydroxypropanamide.

In fig. Figure 5 shows the results of a DSC experiment for crystalline form IIA 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro -2-hydroxypropanamide.

In fig. Figure 6 shows the results of a TGA experiment for crystalline form IIA 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro -2-hydroxypropanamide.

In fig. 7 shows the XRPD pattern of crystalline form IIIA 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro- 2-hydroxypropanamide.

In fig. Figure 8 shows the results of a DSC experiment for crystalline form IIIA 2-(3-(8-amino6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro- 2-hydroxypropanamide.

In fig. Figure 9 shows the results of a TGA experiment for crystalline form IIIA 2-(3-(8-amino6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro- 2-hydroxypropanamide.

In fig. 10 shows the XRPD pattern of crystalline form III of 8-amino-N-(2-hydroxy-2methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo [1,2-a]pyrazine-6-carboxamide.

In fig. 11 shows the results of a DSC experiment for crystalline form III of 8-amino-N-(2hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl )imidazo[1,2-a]pyrazine-6-carboxamide.

In fig. 12 shows the XRPD pattern of crystal form IIB 8-amino-N-(2-hydroxy-2methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo [1,2-a]pyrazine-6-carboxamide.

In fig. 13 shows the results of a DSC experiment for crystalline form IIB 8-amino-N-(2hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl )imidazo[1,2-a]pyrazine-6-carboxamide.

In fig. 14 shows the XRPD pattern of the crystal form IC 8-amino-N-(2-hydroxy-2methylpropyl)-3-(2-(methyl-th ₃ )-5-( 1,1,1-trifluoro-2-hydroxypropane-2- yl)phenyl)imidazo[1,2-a]pyrazine-6carboxamide.

In fig. Figure 15 shows the results of a DSC experiment for the crystalline form IC 8-amino-N-(2hydroxy-2-methylpropyl)-3-(2-(methyl-b ₃ )-5-(1,1,1-trifluoro-2-hydroxypropane- 2-yl)phenyl)imidazo[1,2a]pyrazine-6-carboxamide.

In fig. 16A shows the asymmetric crystalline group of the hydrobromic acid salt 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro -2-hydroxypropanamide, in the form of methanol solvate, with thermal ellipsoids drawn at a 30% probability level.

In fig. 16B shows the crystalline group of the hydrobromic acid salt 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2 -hydroxypropanamide with thermal ellipsoids drawn at a 30% probability level.

Detailed description of the invention

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

which are used, for example, in the preparation of solid dosage forms of the above compound for the treatment of various diseases, including malignant neoplasms.

The present invention also relates to, inter alia, crystalline forms of the PI3K inhibitor 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropane -2-yl)phenyl)imidazo[1,2-a]pyrazin-6-carboxamide having formula (B)

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

I nh ₂ ^HO %N Η, -Ν o γ

F,C £j he (C)

Typically, different crystalline forms of the same substance have different bulk properties, such as hygroscopicity, solubility, stability, etc. Forms with high melting points often have good thermodynamic stability, which is an advantage for extending the shelf life of drugs containing the solid form. Forms with lower melting points are often less thermodynamically stable, but have the advantage of increased solubility in water, resulting in increased bioavailability of the drug. Slightly hygroscopic forms are desirable because of their resistance to heat and humidity and their resistance to degradation during long-term storage. Anhydrous forms are often desirable because they can be prepared in a uniform form without concern for changes in weight or composition due to changes in solvent or water content. On the other hand, hydrated or solvated forms may have the advantage of being less likely to become hygroscopic, and they may exhibit improved resistance to moisture under storage conditions.

As used herein, the term crystalline form is intended to refer to a specific lattice configuration of a crystalline substance. Different crystalline forms of the same substance usually have different crystal lattices (e.g., unit cells), due to their different physical properties characteristic of each crystalline form. In some cases, different lattice configurations have different water or solvent contents. Different crystal lattices can be identified by solid state characterization techniques such as X-ray powder diffractometry (XRPD). Other characterization techniques such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), solid state NMR, etc. further help in identifying the crystalline form and also help in determining the stability and solvent content /water.

Crystalline forms of the substance include both solvated (eg, hydrated) and non-solvated (eg, anhydrous) forms. The hydrated form is a crystalline form that contains water in the crystal lattice. Hydrated forms can be stoichiometric hydrates, in which water is present in the lattice in a certain water/molecule ratio, such as hemihydrates, monohydrates, dihydrates, etc. Hydrated forms can also be non-stoichiometric, in which the water content varies and depends on external conditions , such as humidity.

The characteristics of crystalline forms are usually determined by XRPD. The pattern of XRPD reflections (peaks) is generally considered characteristic of a particular crystal form. It is well known that the relative intensities of XRPD peaks can vary widely depending on, among other things, sample preparation technique, crystal size distribution, filters, sample setup procedure, and the specific instrument used. In some cases, new peaks may be observed, or existing peaks may disappear, depending on the type of equipment or settings (for example, whether a Ni filter is used or not). As used herein, the term peak refers to a reflection having a relative height/intensity of at least approximately 4% of the maximum height/intensity of the peak. Moreover, instrument variations and other factors may affect 2-theta values. Thus, the distributions

- 3,044,976 peaks such as those specified herein may vary by plus or minus about 0.2° (2-theta), and the term essentially as used herein in the context of XRPD is intended to cover the above variations.

Likewise, temperature readings due to DSC, TGA or other thermal experiments may vary by approximately ±4°C depending on the instrument, specific settings, sample preparation, etc. For example, with DSC it is known that observed temperatures will vary on the rate of temperature change, as well as on the method of sample preparation and the specific instrument used. Therefore, the values given in this document relating to DSC thermograms may vary, as stated above, by ± 4°C. Accordingly, the crystalline form provided herein has a DSC thermogram substantially as shown in either figure and should be understood to account for such variation.

Crystalline forms of the compound of formula A.

The compound of formula A can be isolated in many crystalline forms, including, for example, crystalline forms that are anhydrous and/or unsolvated or solvated. In some embodiments, the crystalline forms of the compound of formula A are solvated. In some embodiments, the crystalline forms of the compound of formula A are anhydrous. In some embodiments, the crystalline forms of the compound of formula A are unsolvated. In some embodiments, the crystalline forms of the compound of Formula A are anhydrous and unsolvated. By the term anhydrous it is meant that the crystalline form of the compound of Formula A is substantially free of bound water in the crystal lattice structure, i.e. the compound does not form a crystalline hydrate.

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

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

In some embodiments, the solvent comprises methanol.

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

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

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

This application further provides a crystalline form of the compound 2-(3-(8-amino6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2 -hydroxypropanamide, which is obtained in accordance with the process 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.

In some embodiments, this application provides a process for preparing the hydrobromic acid salt of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)4-methylphenyl)-3, 3,3-trifluoro-2-hydroxypropanamide. In some embodiments, the process includes dissolving a compound of Formula A in a solvent to form a mixture, and adding hydrobromic acid to the mixture.

In some embodiments of the hydrobromic acid salt production process, the solvent comprises methanol.

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

In some embodiments of the process for preparing the hydrobromic acid salt, an excess amount of hydrobromic acid is added to the mixture per 1 eq. 2-(3-(8amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide.

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

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

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

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

In some embodiments, the present application further provides 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)3,3 hydrobromic acid salt. 3-trifluoro-2-hydroxypropanamide, which is obtained in accordance with the process described herein. In some embodiments, the hydrobromic acid salt is crystalline. In some embodiments, the hydrobromic acid salt is a solvated crystalline form. In some embodiments, the hydrobromic acid salt is a crystalline form of methanol solvate.

In some embodiments, the crystalline forms of the invention are substantially isolated. The term substantially isolated means that the particular crystalline form of the compound of formula A is at least partially separated from impurities. For example, in some embodiments, a crystalline form of the invention contains less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Impurities generally include anything that is not essentially an isolated crystalline form, including, for example, other crystalline forms and other substances.

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 a compound of formula A contains more than about 80%, more than about 90%, more than about 95%, more than about 98%, more than about 99%, or more than about 99.5% by weight of a particular crystalline form.

Crystal form IA of the compound of formula A.

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

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

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

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

In some embodiments, crystalline Form IA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 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° ±

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

In some embodiments, Form 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 Form IA of the compound of Formula A has a DSC thermogram substantially as shown in FIG. 2.

In some embodiments, crystalline Form IA of the compound of Formula A has substantially the same footprint on the TGA as shown in FIG. 3.

Crystal form IIA of the compound of formula A.

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

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

In some embodiments, crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 9.1 ± 0.2°. In some embodiments, crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 11.1° ± 0.2°. In some embodiments, crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 21.9° ± 0.2°. In some embodiments, crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 12.6° ± 0.2°. In some embodiments, crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 13.6° ± 0.2°. In some embodiments, crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 18.0° ± 0.2°. In some embodiments, crystalline form IIA of the compound of formula A is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 19.0° ±0.2°.

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

In some embodiments, crystalline Form IIA of the compound of formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 2θ: 9.1° ± 0.2°; 11.1° ± 0.2°; 12.6° ± 0.2°; 13.5° ± 0.2°; 18.0° ± 0.2°; 19.0° ± 0.2°; 20.5° ± 0.2° and 21.9° ± 0.2°. In some embodiments, crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 2θ: 9.1° ±0.2°; 11.1° ±0.2°; 12.6° ± 0.2°; 13.5° ±0.2°; 16.1° ±0.2°; 16.9° ±0.2°; 18.0° ± 0.2°; 18.4° ± 0.2°; 19.0° ± 0.2°; 19.7° ± 0.2°; 20.1° ± 0.2°; 20.5° ± 0.2°; 21.9° ± 0.2°; 23.7° ± 0.2°; 23.8° ± 0.2°; 25.1° ± 0.2°; 25.3° ± 0.2°; 25.8° ± 0.2° and 27.3° ± 0.2°. In some embodiments, crystalline Form IIA of the compound of Formula A has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 2θ: 12.6° ± 0.2°; 18.0° ± 0.2°; 19.0° ± 0.2°; 20.5° ± 0.2° and 21.9° ± 0.2°.

In some embodiments, Form 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 Form IIA of the compound of Formula A has a DSC thermogram substantially as shown in FIG. 5.

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In some embodiments, crystalline Form IIA of the compound of formula A has substantially the same footprint on the TGA as shown in FIG. 6.

Crystal form IIIA of the compound of formula A.

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

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

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

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

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

In some embodiments, Form IIIA of a 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 Form IIIA of the compound of Formula A has a DSC thermogram substantially as shown in FIG. 8.

In some embodiments, crystalline form IIIA of the compound of formula A has substantially the same footprint on the TGA as shown in FIG. 9.

Hydrobromic acid salt of formula A.

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

In some embodiments, the hydrobromic acid salt of the compound of Formula A is crystalline. In some embodiments, the hydrobromic acid salt of the compound of Formula A is a solvated crystalline form. In some embodiments, the hydrobromic acid salt of the compound of formula A is a crystalline methanol solvate form.

Crystalline forms of the compound of formula B.

The compound of formula B can be isolated in many crystalline forms, including, for example, crystalline forms that are anhydrous and/or unsolvated. In some embodiments, the crystalline forms of the compound of Formula B are anhydrous. IN

- 7 044976 In some embodiments, the crystalline forms of the compound of formula B are unsolvated. In some embodiments, the crystalline forms of the compound of Formula B are anhydrous and unsolvated. By the term anhydrous it is meant that the crystalline form of the compound of formula B contains substantially no bound water in the crystal lattice structure, i.e. the compound does not form a crystalline hydrate.

In some embodiments, the crystalline forms provided herein can be prepared, for example, by a process comprising dissolving 8-amino-K-(2hydroxy-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 crystallize the compound from the mixture.

In some embodiments, the process further includes heating the mixture to a temperature of from about 70°C to about 90°C. In some embodiments, the process further includes cooling the mixture to room temperature.

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

In some embodiments, this application provides a crystalline form of the compound 8-amino-K-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2yl )phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, which is prepared in accordance with 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.

In some embodiments, the crystalline forms of the invention are substantially isolated. The term essentially isolated means that the particular crystalline form of the compound of formula B is at least partially separated from impurities. For example, in some embodiments, a crystalline form of the invention contains less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Impurities generally include anything that is not essentially an isolated crystalline form, including, for example, other crystalline forms and other substances.

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 a compound of formula B contains more than about 80%, more than about 90%, more than about 95%, more than about 98%, more than about 99%, or more than about 99.5% by weight of a particular crystalline form.

Crystal form IB of the compound of formula B.

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

The IB crystalline form of a compound of Formula B can be identified by unique signatures, such as X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the IB crystalline form of the compound of Formula B has an XRPD pattern substantially as shown in FIG. 10. The peaks of the XRPD pattern are listed in Table. 4.

In some embodiments, the IB crystalline form of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 6.2° ± 0.2°. In some embodiments, the IB crystalline form of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 15.6° ± 0.2°. In some embodiments, the IB crystalline form of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 20.7° ± 0.2°. In some embodiments, the IB crystalline form of the compound of formula B has an XRPD pattern including the following peaks expressed in degrees 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 IB crystalline form of the compound of Formula B has an XRPD pattern including the following peaks expressed in degrees 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 IB crystalline form of the compound of Formula B has an XRPD pattern including the following peaks expressed in degrees 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 IB crystalline form of the compound of formula B has an XRPD pattern including the following peaks expressed in degrees 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°.

In some embodiments, the IB crystalline form of a compound of formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in

- 8 044976 degrees 2θ: 20: 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 IB crystalline form of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 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 IB crystalline form of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 2θ: 6.2° ± 0.2°; 15.6° ± 0.2°; 16.0° ± 0.2°; 16.7° ± 0.2°; 20.7° ± 0.2° and 23.2° ± 0.2°. In some embodiments, the IB crystalline form of the compound of Formula B has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 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°.

In some embodiments, Form IB of a compound of Formula B is characterized by a DSC thermogram containing an endothermic peak peaking at approximately 174°C. In some embodiments, the Form IB crystalline compound of Formula B has a DSC thermogram substantially as shown in FIG. eleven.

Crystalline form IIB of the compound of formula B.

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

The crystalline form IIB of a compound of formula B can be identified by unique signatures, for example, by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, crystalline form IIB of the compound of formula B has an XRPD pattern substantially as shown in FIG. 12. The peaks of the XRPD pattern are listed in Table. 5.

In some embodiments, crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 4.2° ± 0.2°. In some embodiments, crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 7.4° ± 0.2°. In some embodiments, crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 13.3° ± 0.2°. In some embodiments, crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 20.1° ± 0.2°. In some embodiments, crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 17.0° ± 0.2°. In some embodiments, crystalline form IIB of the compound of formula B is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 18.8° ± 0.2°.

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

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

In some embodiments, Form 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 Form IIB of the compound of Formula B has a DSC thermogram substantially as shown in FIG. 13.

Crystalline forms of the compound of formula C.

The compound of formula C can be isolated in many crystalline forms, including, for example, crystalline forms that are anhydrous and/or unsolvated. Somehow

- 9 044976 In some embodiments, the crystalline forms of the compound of formula C are anhydrous. In some embodiments, the crystalline forms of the compound of formula C are unsolvated. In some embodiments, the crystalline forms of the compound of Formula C are anhydrous and unsolvated. By the term anhydrous it is meant that the crystalline form of the compound of formula C contains substantially no bound water in the crystal lattice structure, i.e. the compound does not form a crystalline hydrate.

In some embodiments, this application provides a process for preparing a crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ₃ )-5-(1,1,1- trifluoro-2hydroxypropan-2-yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide. In some embodiments, the process includes dissolving 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ₃ )-5-(1,1,1trifluoro-2-hydroxypropane-2- yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide in a solvent to form a mixture and crystallize the compound from the mixture.

In some embodiments, the process further includes heating the mixture to a temperature of from about 70°C to about 90°C. In some embodiments, the process further includes cooling the mixture to room temperature.

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

This application additionally provides a crystalline form of the compound 8-amino-X-(2hydroxy-2-methylpropyl)-3-(2-(methyl-d ₃ )-5 -(1,1,1-trifluoro-2-hydroxypropane-2 -yl)phenyl)imidazo[1,2a]pyrazine-6-carboxamide, which is prepared in accordance with the process described herein. In some embodiments, the crystalline form is the IC form described herein.

In some embodiments, the crystalline forms of the invention are substantially isolated. The term essentially isolated means that the particular crystalline form of the compound of formula C is at least partially separated from impurities. For example, in some embodiments, a crystalline form of the invention contains less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 2.5%, less than about 1%, or less than about 0.5% impurities. Impurities generally include anything that is not essentially an isolated crystalline form, including, for example, other crystalline forms and other substances.

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 a compound of formula C contains more than about 80%, more than about 90%, more than about 95%, more than about 98%, more than about 99%, or more than about 99.5% by weight of a particular crystalline form.

Crystalline form IC of compound of formula C.

In some embodiments, the crystalline form of the compound of Formula C is the IC crystalline form. In some embodiments, the IC crystalline form of the compound of formula C is anhydrous and unsolvated. The preparation of the compound of formula C is described in Example 9. The IC crystalline form of the compound of Formula C can generally be prepared as described in Example 10.

The IC crystalline form of a compound of Formula C can be identified by unique signatures, such as X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). In some embodiments, the IC crystalline form of the compound of Formula C has an XRPD pattern substantially as shown in FIG. 14. The peaks of the XRPD pattern are listed in Table. 6.

In some embodiments, the IC crystalline form of the compound of formula C is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 6.2° ± 0.2°. In some embodiments, the IC crystalline form of the compound of formula C is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 11.9° ± 0.2°. In some embodiments, the IC crystalline form of the compound of formula C is characterized by an XRPD pattern including a peak, expressed in degrees 2θ, at 16.7° ± 0.2°. In some embodiments, the IC crystalline form of the compound of formula C has an XRPD pattern including the following peaks expressed in degrees 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 IC crystalline form of the compound of formula C has an XRPD pattern including the following peaks expressed in degrees 2θ: 6.2° ± 0.2°; 15.6° ± 0.2°; 16.0° ± 0.2°; 16.7° ± 0.2° and 21.2° ± 0.2°. In some embodiments, the IC crystalline form of the compound of formula C has an XRPD pattern including the following peaks expressed in degrees 2θ: 6.2° ± 0.2°; 10.4° ± 0.2°; 11.3° ± 0.2°; 11.9° ±0.2° and 12.5° ±0.2°.

In some embodiments, the IC crystalline form of a compound of formula C has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks, expressed in

- 10 044976 degrees 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 IC crystalline form of the compound of Formula C has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 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 IC crystalline form of the compound of Formula C has an XRPD pattern comprising 2 or more, 3 or more, or 4 or more of the following peaks expressed in degrees 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°.

In some embodiments, the IC form of a compound of Formula C is characterized by a DSC thermogram containing an endothermic peak peaking at approximately 179°C. In some embodiments, the IC crystalline form of the compound of Formula C has a DSC thermogram substantially as shown in FIG. 15.

Methods of application

The compounds (eg, salts and crystalline forms) of the invention described herein inhibit PI3Ky kinase activity. Accordingly, the salts and crystalline forms of the invention described herein can be used in methods of inhibiting PI3Ky kinase by contacting the kinase. In some embodiments, the salts and crystalline forms of the invention can be used in methods of inhibiting PI3Ky activity in an individual/patient in need of inhibition by administering an effective amount of the salt or crystalline form described herein. In some embodiments, the modulation is inhibition. In some embodiments, contacting occurs in vivo. In some embodiments, contacting occurs ex vivo. The predominantly crystalline forms described herein demonstrate superior efficacy and favorable safety and toxicity profiles in animal studies.

In some embodiments, PI3Ky contains a mutation. A mutation may be a substitution of one amino acid for another or a deletion of one or more amino acids. In such embodiments, the mutation may be present in the kinase domain of PI3Ky.

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

The salts and crystalline forms described herein may be selective. The term selective means that the salt or crystal form binds to or inhibits PI3Ky with greater affinity or activity, respectively, than at least one other kinase. In some embodiments, the salts and crystalline forms of the invention are selective inhibitors of PI3Ky over PI3Kδ, PI3Ka, and PBKv. In some embodiments, the salts and crystalline forms of the invention are selective inhibitors of PI3Ky over PI3Ka and PBKv. In some embodiments, the selectivity over PI3Kδ can be at least about 2-fold, 3-fold, 5-fold, 10-fold, or 20-fold higher, based on the results of the assay described herein. In some embodiments, the selectivity of each enzyme can be analyzed at a concentration of 2 μM ATP. In some embodiments, the selectivity of the salts and crystalline forms of the invention can be determined using cellular assays associated with specific PI3K kinase activity.

Another aspect of the present invention relates to methods of treating a disease or disorder associated with PI3Ky kinase in an individual (e.g., a patient) by administering to the individual in need of such treatment a therapeutically effective amount or dose of one or more salts or crystalline forms of the present invention or pharmaceutical composition. A disease or disorder associated with PI3Ky may include any disease, disorder or condition that is directly or indirectly associated with the expression or activity of PI3Ky, including levels of overexpression and/or abnormal activity.

In some embodiments, the disease or disorder is an autoimmune disease or disorder, cancer, cardiovascular disease, or neurodegenerative disease.

In some embodiments, the disease or disorder is lung cancer (eg, 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, stomach cancer or sarcoma. In some embodiments, the sarcoma is an Askin's tumor, pampiniform sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft tissue sarcoma, angiosarcoma, phyllodes cystosarcoma, dermatofibrosarcoma protuberans, desmoid tumor, desmoplastic small round cell ocular tumor, epithe- 11 044976 lymphoid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, hemangiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, slave domyosarcoma, synovial sarcoma or undifferentiated pleomorphic sarcoma.

In some embodiments, the disease or disorder is mesothelioma or adrenocarcinoma. In some embodiments, the disease or disorder is mesothelioma. In some embodiments, the disease or disorder is adrenocarcinoma.

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

In some embodiments, the disease or disorder is Burkitt's lymphoma, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanthoma, lymphoplasmacytic lymphoma, extranodal marginal cell lymphoma zones, Waldenström's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosal associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large B-cell lymphoma, lymphogranulomatosis, splenic marginal zone lymphoma, primary effusion lymphoma, intravascular B large cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of unknown etiology (MGUE), or diffuse large B-cell lymphoma.

MDSC (myeloid-derived suppressor cells) are a heterogeneous group of immune cells of myeloid origin (a family of cells derived from bone marrow stem cells). MDSC proliferate greatly in pathological situations such as chronic infections and cancer, as a result of altered hematopoiesis. MDSCs differ from other types of myeloid cells in that they have strong 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 regulate their functions. In some embodiments, the compounds, etc., described herein can be used in methods relating to cancer tissue (eg, tumors) with high MDSC infiltration, including solid tumors with high basal macrophage levels and/or MDSC infiltration.

In some embodiments, the disease or disorder is Burkitt's lymphoma, acute myeloid leukemia, chronic myelogenous leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone cell lymphoma, macroglobulin Iu Waldenström, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosal associated lymphoid tissue (MALT) lymphoma, mediastinal (thymic) large cell lymphoma, lymphogranulomatosis, marginal zone lymphoma of the spleen, primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma , smoldering myeloma (also known as asymptomatic myeloma), monoclonal gammopathy of unknown etiology (MGUE), or diffuse large B-cell lymphoma.

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

In some embodiments, the diffuse large B cell lymphoma is diffuse large B cell lymphoma of activated B cell type (ABC) or diffuse large B cell lymphoma of germinal center B cells (GCB).

In some embodiments, Burkitt's lymphoma is endemic Burkitt's lymphoma, sporadic Burkitt's lymphoma, or Burkitt's lymphoma-like lymphoma.

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

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

In some embodiments, the disease or disorder is cardiac hypertrophy, cardiomyocyte dysfunction, chronic obstructive pulmonary disease (COPD), high blood pressure, ischemia, ischemia-reperfusion, vasoconstriction, anemia (e.g., hemolytic anemia, aplastic anemia, or pure red cell 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.

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

In some embodiments, idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP.

In some embodiments, the vasculitis is Behçet's disease, Cogan's syndrome, giant cell arteritis, polymyalgia rheumatica (PPR), Takayasu's arteritis, Buerger's disease (thromboangiitis obliterans), central nervous system vasculitis, Kawasaki disease, polyarteritis nodosa, Churg-Strauss syndrome, mixed cryoglobulinemic vasculitis (essential or caused by hepatitis C virus (HCV)), Henoch-Schönlein purpura (HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener's granulomatosis or antineutrophil cytoplasmic antibody (ANCA)-associated systemic vasculitis (AASV).

The present invention further provides a salt or crystalline form described herein for use in any of the methods described herein.

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

As used herein, the term contacting refers to the association of said moieties in an in vitro system or in an in vivo system. For example, contacting PI3K with a salt or crystalline form of the invention involves administering a crystalline form of the invention to an individual or patient, such as a human, having PI3K, as well as, for example, administering a salt or crystalline form of the invention to a sample containing a cellular or purified drug, containing PI3K.

It is believed that the salts and crystalline forms provided herein, or any embodiments thereof, may have a satisfactory pharmacological profile and promising biopharmaceutical properties, such as toxicological profile, metabolism and pharmacokinetic properties, solubility and permeability. It should be understood that determination of relevant biopharmaceutical properties is within the skill of the art, such as determination of cytotoxicity in cells or inhibition of certain targets or channels to determine potential toxicity.

As used herein, the terms individual or patient, used interchangeably, refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, and most preferably to people.

As used herein, the phrase therapeutically effective amount refers to the amount of active salt, crystalline form, or pharmaceutical agent that produces the biological or drug response desired in a tissue, system, animal, or individual.

- 13 044976 or a human researcher, veterinarian, family doctor or other clinician.

As used herein, the term treating or treating may refer to one or more of (1) inhibiting a disease; for example, inhibiting a disease, condition or disorder in an individual who experiences or exhibits pathological or symptomatic signs of the disease, pathological condition or disorder (ie, stopping further development of pathological and/or symptomatic signs); and (2) weakening of the disease; for example, alleviation of a disease, condition or disorder in an individual who perceives or exhibits pathological or symptomatic signs of the disease, pathological condition or disorder (ie, reversal of the pathological and/or symptomatic signs), such as a decrease in the severity of the disease.

In some embodiments, the salts and crystalline forms of the invention are useful for preventing or reducing the risk of developing any of the diseases mentioned herein; for example, preventing or reducing the risk of developing a disease, condition or disorder in a subject who may be predisposed to the disease, condition or disorder but is not yet experiencing or demonstrating disease pathology or symptoms.

Combined therapeutic agents.

I. Therapeutics that affect immune checkpoints.

In some embodiments, the PI3Ky 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, a combination with one or more immune checkpoint inhibitors, as described herein, can be used to treat melanoma. The salts and crystalline forms of the present invention can be used in combination with one or more immune checkpoint inhibitors. Illustrative examples of immune checkpoint inhibitors include inhibitors targeting 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 checkpoint stimulating molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is a checkpoint inhibitory 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 invention can 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.

In some embodiments, the PI3Ky inhibitors provided herein can be used in combination with one or more immune checkpoint molecule agonists, for example, OX40, CD27, OX40, GITR, and CD137 (also known as 4-1BB).

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

In some embodiments, the immune checkpoint molecule inhibitor is a PD-1 inhibitor, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), durvalumab (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 antibody is MGA012. In some embodiments, the anti-PD1 antibody is SHR-1210. Other antineoplastic agent(s) include therapeutic antibodies such as 4-1BB (eg, urelumab, utomilumab).

In some embodiments, the immune checkpoint molecule inhibitor is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

In some embodiments, the immune checkpoint molecule inhibitor is an inhibitor of PD-1 and PD-L1, such as an anti-PD-1/PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1/PD-L1 antibody is MCLA136.

In some embodiments, the inhibitor is MCLA-145.

- 14 044976

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

CP-675.206.

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

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

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

In some embodiments, the immune checkpoint inhibitor molecule is an OX40 agonist, such as an OX40 antibody or OX40L fusion protein agonist. In some embodiments, the anti-OX40 antibody is MEDI0562, MOXR0916, PF-04518600, GSK3174998, or BMS-986178. In some embodiments, the OX40L fusion protein is MEDI6383.

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

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

In some embodiments, the PI3Ky inhibitors provided herein can be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitor is an IDO1, TDO, or arginase inhibitor. Examples of IDO1 inhibitors include epacadostat, NLG919, BMS986205, PF-06840003, IOM2983, RG-70099 and LY338196.

As provided throughout, additional compounds, inhibitors, agents, etc. may be combined with a compound of the present invention in a single or continuous dosage form, or they may be administered simultaneously or sequentially in separate dosage forms.

II. Therapeutic agents for the treatment of malignant neoplasms.

The growth and survival of malignant cells can be influenced by numerous signaling pathways. Thus, it is effective to combine different inhibitors of enzymes/proteins/receptors that alter the activity of different targets to treat such pathological conditions. Targeting more than one signaling pathway (or more than one biological molecule involved in a given signaling pathway) may reduce the likelihood of drug resistance occurring in a cell population and/or reduce treatment toxicity.

The salts and crystalline forms of the present invention can be used in combination with one or more other enzyme/protein/receptor inhibitors or one or more therapeutic agents for the treatment of diseases such as cancer. Examples of diseases and indications that can be treated with combination therapy include those described herein. Examples of malignancies include solid tumors and liquid tumors such as hematologic malignancies.

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

For example, the salts and crystalline forms disclosed herein may be combined with one or more inhibitors of the following kinases for the treatment of cancers and other diseases or disorders described herein: Akt1, Akt2, Akt3, TGF-PR, PKA, PKG , PKC, CaM kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3,

- 15 044976

HER4, INS-R, IGF-1R, IR-R, PDGFaR, PDGFeR, CSFIR, KIT, FLK-II, KDR/FLK-1, FLK-4, ft-1, FGFR1, FGFR2, FGFR3, FGFR4, c- Met, Ron, Sea, TRKA, TRKB, TRKC, FLT3, VEGFR/Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK and B-Raf. Non-limiting examples of inhibitors that can be combined with the salts and crystalline forms of the present invention for the treatment of cancers and other diseases and disorders described herein include an FGFR inhibitor (FGFR1, FGFR2, FGFR3 or FGFR4, e.g. INCB54828, INCB62079 and INCB63904) , JAK inhibitor (JAK1 and/or JAK2, e.g. ruxolitinib, baricitinib or INCB39110), IDO inhibitor (e.g. epacadostat, NLG919 or BMS986205), LSD1 inhibitor (e.g. INCB59872 and INCB60003), TDO inhibitor, PI3K-delta inhibitor (e.g. , INCB50797 and INCB50465), Pim inhibitor, CSF1R inhibitor, TAM receptor tyrosine kinases (Tyro-3, Axl and Mer), histone deacetylase (HDAC) inhibitor such as HDAC8 inhibitor, angiogenesis inhibitor, interleukin receptor inhibitor, bromine and extra-terminal family member inhibitors (eg, bromine domain inhibitors or BET inhibitors such as INCB54329 and INCB57643) and an adenosine receptor antagonist or combinations thereof.

In some embodiments, the salts and crystalline forms described herein are administered together with a PI3Kδ inhibitor. In some embodiments, the salts and crystalline forms described herein are administered together with a JAK inhibitor. In some embodiments, the salts and crystalline forms described herein are administered together with a JAK1 or JAK2 inhibitor (eg, baricitinib or ruxolitinib). In some embodiments, the salts and crystalline forms described herein are administered together with a JAK1 inhibitor. In some embodiments, the salts and crystalline forms described herein are administered together with a JAK1 inhibitor that is superior to JAK2 in selectivity.

Examples of antibodies for use in combination therapy include, but are not limited to, trastuzumab (eg, anti-HER2), ranibizumab (eg, anti-VEGF-A), bevacizumab (trade name Avastin, eg, anti-VEGF, panitumumab ( eg anti-EGFR), cetuximab (eg anti-EGFR), Rituxan (anti-CD20) and antibodies targeting c-MET.

One or more of the following agents may be used in combination with the salts and crystalline forms of the present invention and are presented as a non-limiting list: cytostatic agent, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epothilones , tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, SCH 66336, R115777, L778,123, BMS 214662, IRESSA™ (gefitinib), TARCEVA™ (erlotinib), anti-EGFR antibodies, intron, ara-C, adriamycin, cytoxan , gemcitabine, uracil mustard, chlormethine, ifosfamide, melphalan, chlorambucil, pipobromane, triethylene melamine, triethylene thiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, o xaliplatin, leucovirin, ELOXATIN ™ (oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17αethinyl estradiol, diethylstilbestrol, testosterone, pred nisone, fluoxymesterone, dromostanolone propionate , testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, amiglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, navelbine, anastrazole, letrazole, capecitabine, reloxafine, droloxafine, hexamethylmelamine, Avastin, HERCEPTIN™ (trastuzumab), BEXXAR™ (tositumomab), VELCADE™ (bortezomib), ZEVALIN™ (ibritumomab tiuxetan), TRISENOX™ (arsenic trioxide), XELODA™ (capecitabine ), vinorelbine, porfimer, ERBITUX™ (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, lerozol, fulvestrant, exemestane, ifosfomide, rituximab, C225 (cetuximab), campath (alemtuzumab), clofarabine, cladribine, aphidicolon, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triapine, didox, trimidox, amidox, 3-AP and MDL-101,731.

The salts and crystalline forms of the present invention may further be used in combination with other treatments for cancer, such as chemotherapy, radiation therapy, targeted tumor therapy, adjuvant therapy, immunotherapy, or surgery. Examples of immunotherapy include cytokine therapy (eg, interferons, GM-CSF, G-CSF, IL-2), immunotherapy using CRS-207, cancer vaccine, monoclonal antibody, T cell adoptive transfer, toll receptor agonists , STING agonists, oncolytic virotherapy and immunomodulatory small molecules, including thalidomide or a JAK1/2 inhibitor, etc. Salts and crystalline forms can be administered in combination with one or more anticancer drugs, such as chemotherapy drugs. Examples of chemotherapy drugs include any of the following: abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, bortezomib, busulfan

- 16 044976 intravenous, busulfan orally, calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, daunorubicin, decitabine, denileukin, denileukin diftitox , dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin ace tat, histrelin acetate, ibritumomab tiuxetan, idarubicin ifosfamide, imatinib mesylate, interferon alfa2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenylpropionate, nelarabine , nofetumomab, olaparib, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobromane, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, rucaparib, streptozocin, tamoxifen, temozolomi d, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, niraparib, veliparib, talazoparib, and zoledronate.

Additional examples of chemotherapeutic agents include proteasome inhibitors (eg, bortezomib), thalidomide, Revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

Examples of steroids include corticosteroids such as dexamethasone or prednisone.

Examples of Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC™), nilotinib, dasatinib, bosutinib and ponatinib and pharmaceutically acceptable salts. Other examples of suitable Bcr-Abl inhibitors include the compounds and their pharmaceutically acceptable salts of the genera and species disclosed in US Patent No. 5,521,184, WO 04/005281 and US Patent Serial No. 60/578,491.

Examples of suitable Flt-3 inhibitors include midostaurin, lestaurtinib, linifanib, sunitinib, sunitinib, maleate, sorafenib, quizartinib, crenolanib, pacritinib, tandutinib, PLX3397 and ASP2215 and pharmaceutically acceptable salts thereof. Another example of suitable Flt-3 inhibitors includes the compounds and their pharmaceutically acceptable salts disclosed in WO 03/037347, WO 03/099771 and WO 04/046120.

Examples of suitable RAF inhibitors include dabrafenib, sorafenib and vemurafenib, as well as pharmaceutically acceptable salts thereof. Another example of suitable RAF inhibitors includes the compounds and their pharmaceutically acceptable salts as described in WO 00/09495 and WO 05/028444.

An example of suitable FAK inhibitors includes VS-4718, VS-5095, VS-6062, VS-6063, BI853520 and GSK2256098, and pharmaceutically acceptable salts thereof. Another example of suitable FAK inhibitors includes the compounds and their pharmaceutically acceptable salts as described in WO 04/080980, WO 04/056786, WO 03/024967, WO 01/064655, WO 00/053595 and WO 01/014402.

In some embodiments, the salts and crystalline forms of this invention can be used in combination with one or more other kinase inhibitors, including imatinib, especially for the treatment of patients resistant to imatinib or other kinase inhibitors.

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

In some embodiments, the salts and crystalline forms of this invention can be used in combination with a JAK or PI3Kδ inhibitor.

The agents can be combined with the present compound in a single or continuous dosage form, or the agents can be administered simultaneously or sequentially in separate dosage forms.

The salts and crystalline forms of the present invention can be used in combination with

- 17 044976 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.

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

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

The salts and crystalline forms of the present invention described herein can be used in combination with a vaccination protocol for the treatment of malignancy. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins from viruses implicated in human cancer, such as human papillomaviruses (HPV), hepatitis viruses (HBV and HCV) ) and Kaposi's sarcoma herpes virus (KHSV). In some embodiments, the salts and crystalline forms of the present invention can be used in combination with a tumor-specific antigen, such as heat shock proteins isolated from the tumor tissue itself. In some embodiments, the salts and crystalline forms can be combined with immunization of dendritic cells to activate potent antitumor responses.

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

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

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 such a therapeutic approach may be particularly suitable include pathogens for which there is currently no effective vaccine, or pathogens for which conventional vaccines are not completely effective. These include, but are not limited to, HIV, hepatitis (A, B and C), influenza, herpes, giardia, malaria, leishmania, Staphylococcus aureus, Pseudomonas aeruginosa.

Viruses causing infections that can be treated using the methods of the present invention include, but are not limited to, human papillomavirus, influenza virus, hepatitis A, B, C or D viruses, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory syndrome, Ebola virus, measles virus, herpes virus (for example, VZV, HSV-1, HAV-6, HSV-II and CMV, Epstein-Barr virus), flaviviruses, ECHO virus, rhinovirus, Coxsackie virus, coronavirus, respiratory Syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papilloma virus, shellfish virus, polio virus, rabies virus, JC virus and arboviral encephalitis virus.

Pathogenic bacteria causing infections that can be treated using the methods of the present invention include, but are not limited to, chlamydia, rickettsial bacteria, mycobacteria, staphylococci, streptococci, pneumococci, meningococci and conococci, klebsiella, proteus, serratia, pseudomonas, legionella, diphtheria, salmonella, bacilli, bacteria that cause cholera, tetanus, botulism, anthrax, plague, leptospirosis and Lyme disease.

Pathogenic fungi causing infections that can be treated using the methods of the present invention include, but are not limited to, Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.) etc.), genus Mucorales (mucor, absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis and Histoplasma capsulatum. Pathogenic parasites causing infections that can be treated using the methods of the present invention include, but are not limited to, Entamoeba histolytica, Balantidium coli, Naegleriafowleri, Acanthamoeba spp., Giardia lambia, Cryptosporidium spp., Pneumocystis carinri, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani,

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Toxoplasma gondi and Nippostrongylus brasiliensis.

Methods for safely and effectively administering most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in the standard literature. For example, the administration of many of the chemotherapeutic agents is described in the Physicians' Desk Reference (PDR, eg, 1996 edition, Medical Economics Company, Montvale, NJ), the description of which is incorporated herein by reference as if set forth in its entirety.

Pharmaceutical compounds and dosage forms.

When the compounds of the invention (eg, salts and crystalline forms) are used as pharmaceuticals, they can be administered in the form of pharmaceutical compositions. These compositions can be prepared using methods well known in the art of pharmacy and can be administered in a variety of ways depending on whether local or systemic treatment is desired and the area being treated. Administration may be local (including transdermal, epidermal, ocular and mucosal, including intranasal, vaginal and rectal delivery), pulmonary (eg, by inhalation or insufflation of powders or aerosols, such as a nebulizer; intratracheal or intranasal) , oral or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion; or intracranial, such as intrathecal or intraventricular administration. Parenteral administration can be carried out as a single bolus dose or, for example, continuously using a perfusion pump. Pharmaceutical and topical compositions may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

This invention also includes pharmaceutical compositions that contain as an active ingredient a compound herein or a pharmaceutically acceptable salt thereof in combination with one or more pharmaceutically acceptable excipients. In some embodiments of the invention, the composition is suitable for topical use. To prepare the compositions of the present invention, the active ingredient is typically mixed with an excipient, diluted with an excipient, or included in a carrier in the form of, for example, a capsule, sachet, paper bag, or other container. When the excipient serves as a diluent, it may be a solid, semi-solid, or liquid material that serves as a carrier, carrier, or vehicle for the active ingredient. Thus, the compositions can be in the form of tablets, dragees, powders, lozenges, sachets, starch capsules, infusions, suspensions, emulsions, solutions, syrups, aerosols (solid or liquid), ointments containing, for example, up to 10 wt. % active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions or sterile packaged powders.

To prepare the formulation, the active compound may be milled to provide a suitable particle size before combining with other ingredients. If the active compound is substantially insoluble, it can be milled to obtain particles less than 200 mesh in size. If the active compound is substantially water soluble, the particle size can be adjusted by grinding to provide substantially uniform distribution in the formulation, for example about 40 mesh.

The compounds of the present invention can be milled using known milling methods, such as wet milling, to obtain particles of a size suitable for the formation of tablets and other types of formulations. Finely divided (nanoparticle) preparations of the compounds herein can be prepared using methods known in the art, for example, see International Application No. WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose. The compositions may further contain: lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl and propyl hydroxybenzoate; sweeteners; and flavors. The compositions of the present invention can be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a patient using methods known in the art.

The compositions may be formulated in unit dosage form, with each dose containing from about 5 to about 1000 mg (1 g), typically from about 100 to about 500 mg, of the active ingredient. The term unit dosage forms refers to physically discrete units suitable as unit doses for humans and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect, in combination with a suitable pharmaceutical excipient.

In some embodiments, the compositions of the present invention contain from about

- 19 044976 up to about 50 mg of active ingredient. One of ordinary skill in the art will appreciate that this includes compositions containing from about 5 to about 10, from about 10 to about 15, from 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 active ingredient.

In some embodiments, the compositions of the present invention contain from about 50 to about 500 mg of active ingredient. One of ordinary skill in the art will appreciate that it includes compositions containing from about 50 to about 100, from about 100 to about 150, from about 150 to about 200, from about 200 to about 250, from about 250 to about 300, from about 350 to about 400 or about 450 to about 500 mg of active ingredient.

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

Similar dosages may be used for the compounds described herein in the methods and uses of the present invention.

The active compound can be effective over a wide range of dosages and is typically administered in a pharmaceutically effective amount. It is understood, however, that the amount of compound that is actually administered will generally be determined by the physician in accordance with the circumstances at hand, including the condition being treated, the route of administration selected, the particular compound administered, the age, weight and response of the particular patient, the severity of the patient's symptoms, and the like.

To prepare solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient to obtain a solid preformulation containing a homogeneous mixture of the compound of the present invention. When these preformulations are stated to be homogeneous, it is understood that the active ingredient is generally dispersed uniformly throughout the composition so that the composition can be readily divided into equally effective unit dosage forms such as tablets, dragees and capsules. This solid preformulation is then divided into unit dosage forms of the types presented above, containing from, for example, about 0.1 to about 1000 mg of the active ingredient of the present invention.

The tablets or dragees of the present invention may be coated or otherwise modified to provide a dosage form having the advantage of a sustained release. For example, a tablet or dragee may comprise an inner dosage component and an outer dosage component, the latter in the form of a coating for the former. The two components can be separated by an enteric layer, which prevents degradation in the stomach and allows the internal component to enter the duodenum intact or be released in a delayed manner. A variety of materials can be used for such enteric layers or coatings, such materials containing a number of polymer acids and mixtures of polymer acids with materials such as shellac, cetyl alcohol and cellulose acetate.

Liquid forms into which the compounds and compositions of the present invention may be formulated, for oral administration or injection, include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil , coconut oil or peanut butter, as well as infusions and similar pharmaceutical media.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients described above. In some embodiments, the compositions are administered into the respiratory tract through the mouth or nose for local or systemic effect. The compositions can be nebulized using inert gases. Nebulized solutions can be inhaled directly from the nebulization device, or the nebulization device can be attached to a face mask, oxygen tent, or intermittent positive pressure breathing apparatus. Solutions, suspensions or powder compositions can be administered orally or nasally using devices that deliver the drug in an appropriate manner.

Formulations for topical use 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, and the like. Carriers for cream compositions may be based on water in combination with glycerin and one or more other components, for example glyceryl monostearate, PEG-glyceryl monostearate and cetyl stearyl alcohol. Gels can be prepared using isopropyl alcohol and water, preferably in combination

- 20 044976 with other components, for example glycerin, hydroxyethylcellulose and the like. In some embodiments, the 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 compound of the present invention. Topical formulations may be packaged in suitable tubes, for example 100 g, which may optionally contain instructions for treating a selected symptom, such as psoriasis or other skin condition.

The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration, such as prophylaxis or therapy, the patient's condition, route of administration, and the like. For therapeutic purposes, the compositions can be administered to a patient already suffering from a disease in an amount sufficient to treat or at least partially alleviate the symptoms of the disease or its complications. Effective dosages will depend on the condition being treated as well as the judgment of the attending physician based on factors such as the severity of the disease, the age, weight and general condition of the patient, and the like.

Compositions administered to a patient may be in the form of pharmaceutical compositions described above. These compositions can be sterilized by conventional sterilization methods or can be sterile filtered. Aqueous solutions may be packaged for use as is or lyophilized, the lyophilized formulation being combined with a sterile aqueous vehicle prior to administration. The pH of compound formulations is typically from 3 to 11, more preferably from 5 to 9, and most preferably from 7 to 8. It should be understood that the use of some of the above excipients, carriers or stabilizers will result in the formation of pharmaceutical salts.

The therapeutic dosage of a compound of the present invention may vary according to, for example, the specific use being treated, the route of administration of the compound, the health and pathological condition of the patient, and the judgment of the attending physician. The proportion or concentration of a compound herein in a pharmaceutical composition may vary depending on a number of factors, including dosage, chemical characteristics (eg, hydrophobicity), and route of administration. For example, the compounds of the present invention may be present in an aqueous physiological buffer solution containing from about 0.1 to about 10% w/v of the compound for parenteral administration. Some typical doses range from about 1 mcg/kg to about 1 g/kg body weight per day. In some embodiments, the dose ranges from about 0.01 mg/kg to about 100 mg/kg body weight per day. Dosage will likely depend on variables such as the type and degree of progression of the disease or disorder, the general health of the individual patient, the relative biological effectiveness of the selected compound, the composition of the excipient, and the route of administration. Effective doses can be extrapolated from dose-response curves obtained from in vitro test systems or animal models.

The compositions of the present invention may further include one or more additional pharmaceutical agents, such as a chemotherapeutic, steroid, anti-inflammatory compound or immunosuppressant, examples of which are listed herein.

Labeled compounds and methods of analysis.

Another aspect of this disclosure relates to labeled compounds (eg, salts and crystalline forms) of the invention (radiolabeled, fluorescently labeled, etc.) that would be useful not only in imaging techniques, but also in assays such as in vitro , and in vivo, to detect and quantify PI3K in tissue samples, including human tissue, and to identify PI3K ligands by inhibition of labeled compound binding. Substitution of one or more atoms of compounds described herein may also be useful in creating differentiated ADME (Adsorption, Distribution, Metabolism, and Excretion). Accordingly, the present disclosure includes PI3K assays that contain such labeled or substituted compounds.

The present invention further includes isotopically labeled compounds of the present invention. An isotopically or radioactively labeled compound is a compound of the invention in which one or more atoms are replaced or replaced by an atom 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 included in the compounds of the present invention include, but are not limited to, ² H (also referred to as D for deuterium), ³ H (also referred to as T for tritium), ⁿ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ¹⁸ F, ³⁵ S, ³⁶ Cl, ⁸² Br, ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I and ¹³¹ I. For example, one or more The hydrogen atoms in the compound of the present invention may be replaced by deuterium atoms (eg, one or more hydrogen atoms of the C ₁ . ₆ α-alkyl group of formula (I) may optionally be replaced by deuterium atoms, eg -CD3 may serve as a substitute for -CH3). In some embodiments, the alkyl groups of the disclosed formulas and/or forms may be perdeuterated.

One or more of the constituent atoms of the compounds presented herein may be

- 21 044976 replaced or replaced by isotopes of atoms in natural or non-natural quantities. In some embodiments, the compound contains at least one deuterium atom. For example, one or more hydrogen atoms in a compound provided herein may be replaced or replaced by deuterium (eg, one or more hydrogen atoms of a C ₁ . ₆ alkyl group may be replaced by deuterium atoms, eg -CD3 resulting from -CH3 substitution). In some embodiments of the invention, 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 of the hydrogen atoms in the compound may be replaced or replaced by deuterium atoms.

In some embodiments, each of the 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms attached to the carbon atoms of any Formula A substituents is optionally substituted with a deuterium atom.

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

Substitution with heavier isotopes such as deuterium may provide certain therapeutic benefits due to greater metabolic stability, such as increased in vivo half-life or the need for lower dosages, and may therefore be preferable in some circumstances (see e.g. A Kerekes et al J Med Chem 2011, 54, 201-210 R Xu et al J Label Compd Radiopharm 2015, 58 308-312). In particular, substitution at one or more metabolic sites may provide one or more therapeutic benefits.

The radionuclide that is included in the present radiolabeled compounds will depend on the particular application of the radiolabeled compound. For example, for in vitro PI3K labeling and competition assays, compounds containing ³ H, ¹⁴ C, ⁸² Br, ¹²⁵ I, ¹³¹ I or ³⁵ S may be suitable. For radioimaging purposes, ⁿ C, ¹⁸ F, ¹²⁵ 1, ¹²³ may be suitable 1, ¹²⁴ 1, ¹³¹ 1, ⁷⁵ Br, ⁷⁶ Br or ⁷⁷ Br.

It is understood that a radiolabeled or labeled compound is a compound that contains at least one radionuclide. In some embodiments, the radionuclide is selected from the group consisting of H, ¹⁴ C, ¹²⁵ 1, ³⁵ S and ⁸² Br.

The present invention may further include synthetic methods for introducing radioisotopes into the compounds of the present invention. Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and those of ordinary skill in the art will readily understand the methods applicable to the compounds of the present invention.

The labeled compound of the present invention can be used in a screening assay for identifying/evaluating compounds. For example, a newly synthesized or identified compound (ie, a test compound) that is labeled can be assessed for its ability to bind PI3K by monitoring the change in its concentration upon contact with PI3K by monitoring labeling. For example, a test compound (labeled) can be assessed for its ability to reduce the binding of another compound known to bind to PI3K (ie, a standard compound). Accordingly, the ability of a test compound to compete with a standard compound for binding to PI3K directly correlates with its binding affinity. Conversely, in some other screening studies the standard compound is labeled but the test compound is not labeled. Accordingly, the concentration of the labeled standard compound is monitored to assess the competition between the standard compound and the test compound and thereby determine the relative binding capacity of the test compound.

Sets.

The present description also includes pharmaceutical kits useful, for example, for the treatment or prevention of PI3K-related diseases or disorders, such as cancer, which include one or more containers containing a pharmaceutical composition containing a therapeutically effective amount of salts and crystalline forms of the invention. Such kits may further contain, if desired, one or more of a variety of components of conventional pharmaceutical kits, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art. technology. Instructions, either in the form of inserts or labels indicating quantities to be administered, administration guides, and/or mixing instructions, may also be included in the kit.

The invention will be described in more detail using specific examples. The following examples are offered for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art will readily recognize many non-critical parameters that can be changed or modified to obtain substantially the same results. The salts and crystalline forms of the Examples have been found to be ΡΙ3Κγ inhibitors according to at least one assay described herein.

Examples

Preparatory LC-MS purifications of some of the obtained compounds were carried out in Waters mass-directed fractionation systems. The basic hardware setup, protocols and control software for operating 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 (LC-MS) to verify purity under the following conditions: instrument; Agilent 1100 series LC/MSD, column: Waters Sunfire™ C18 5 µm, 2.1x50 mm; buffers: mobile phase A: 0.025% TFA in water and mobile phase B: acetonitrile; gradient from 2% to 80% B in 3 min with a flow rate of 2.0 ml/min.

Some of the resulting compounds were also separated on a preparative column using reverse phase high performance liquid chromatography (RP-HPLC) with MS detector or flash chromatography (silica gel) as indicated in the examples. Typical column conditions for preparative reversed-phase high-performance liquid chromatography (RP-HPLC) are as follows:

pH 2 purification: Waters Sunfire™ C ₁₈ 5 µm, 30x100 mm or Waters XBridge™ C ₁₈ 5 µm, 30x100 mm column, eluting with mobile phase A: 0.1% TFA (trifluoroacetic acid) in water and mobile phase B: acetonitrile; the flow rate was 60 mL/min, and the separation gradient 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)).

pH 10 purifications: Waters XBridge™ C ₁₈ 5 µm, 30x100 mm column, eluting 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)).

In the examples below, X-ray powder diffraction analysis was performed on a Bruker D8 Advance ECO X-ray Powder Diffractometer (XRPD) using the following parameters: radiation source: copper at 1.5418 A, LYNXEYETM detector and X-ray power 40 kV, 25 mA . The sample powder was dispersed on a sample holder with zero background. The general measurement conditions were as follows: initial angle, 3°; stopping angle - 30°; sample input - 0.015 degrees; and scanning speed - 2°/min.

Differential scanning calorimetry (DSC) was performed on a TA Instrument Differential Scanning Calorimetry, Discovery DSC2500 with an automatic sampler. General experimental conditions: 20-300°C at 10°C/min, nitrogen gas flow 50 ml/min using an aluminum sample crucible.

Thermogravimetric analysis (TGA) was carried out on a TA Instrument Thermogravimetric Analyzer, TGA5500 with an automatic sampler under the following conditions: Temperature change rate 10°C/min from 25°C to 600°C; purging with nitrogen gas at a rate of 25 ml/min followed by balancing the purge flow; and a platinum crucible for samples.

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

- 23 044976

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

BUT CF ₃

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 in tetrahydrofuran (0.654 ml, 0.654 mmol) at 0°C and stirred for several minutes at 0°C. The ice bath was removed and the resulting reaction mixture was stirred for 1.5 hours. The reaction mixture was again cooled to 0°C and treated with water (6.48 ml, 360 mmol) and 1.0 M tetrabutylammonium fluoride in tetrahydrofuran (6.54 ml , 6.54 mmol). The ice bath was removed and the reaction mixture was stirred at ambient temperature for 30 minutes. The yellow reaction mixture was diluted with brine (150 ml) and extracted with ethyl acetate (200 ml). The organic layer was washed with saturated ammonium chloride (100 ml), dried over sodium sulfate, filtered and concentrated to give a tan oil. Purification by flash column chromatography using methyl tert-butyl ether (MTBE) in hexanes (0% to 50%) provided the desired product (8.42 g, 95.7%) as a yellow oil. LC-MS for C ₉ H ₇ BrF ₃ (M-OH) ⁺ : m/z=251.0, 253.0; found: 250.9, 252.8.

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

A mixture of 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-ol (8.41 g, 31.3 mmol) in dichloromethane (125 ml) at 0°C was treated with Dess-Martin periodinane (19.9 g, 46.9 mmol) and stirred at RT for 2.5 hours. The reaction mixture was concentrated (by rotary evaporation in a water bath set at 30°C) to give an oily solid, which was diluted with diethyl ether ( 200 ml), which resulted in the precipitation of additional solids. This mixture was filtered through Celite®, and washed with Celite® with additional diethyl ether (200 ml). The filtrate was washed with saturated sodium bicarbonate (3x200 ml) and brine, dried over sodium sulfate, filtered and concentrated to give an oily solid. The oily solid was partitioned between diethyl ether (150 ml) and water (100 ml). The organic layer was separated and washed with saturated sodium bicarbonate (2x75 ml) and brine, dried over sodium sulfate, filtered and concentrated to give the desired product (7.93 g, 95.0%) as an oil, which was used without further purification. LC-MS for C ₉ H ₇ BrF ₃ O (M+H) ⁺ : m/z=267.0, 269.0; found: 267.1, 268.9.

Step 3. 2-(3-Bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanenitrile

A solution of 1-(3-bromo-4-methylphenyl)-2,2,2-trifluoroethan-1-one (7.92 g, 29.7 mmol) in dichloromethane (29.7 ml) was treated with trimethylsilyl cyanide (8.70 ml , 65.2 mmol), potassium cyanide (0.290 g, 4.45 mmol) and 18-crown-6 (0.290 g, 1.10 mmol) and stirred for 1 hour. The reaction mixture was cooled in an ice bath due to exothermia after adding 18-crown-6. The reaction mixture was concentrated (on a rotary evaporator at a water bath temperature of 28°C) to obtain a rust-colored solid. The solid was dissolved in THF (29.6 mL), cooled to 0°C, treated with 1.8 M HCl (10.9 mL, 19.6 mmol) and stirred at room temperature (RT) for 1. 5 o'clock. The reaction mixture was diluted with water (75 ml) and extracted with diethyl ether (3x75 ml). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated. Reconcentration from hexanes provided the desired product (8.70 g, 99.8%) as an orange solid, which was used without further purification. LC-MS for C ₉ H ₇ BrF ₃ O (M-CN) ⁺ : m/z=267.0, 269.0; found: 266.9, 269.0.

- 24 044976

Step 4. 2-(3-Bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (second enantiomer to elute)

A solution of 2-(3-bromo-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropannithoryl (8.70 g, 29.6 mmol) in 1,4-dioxane (59.2 ml) at 0° C was treated with concentrated HCl solution (9.00 mL, 108 mmol) pre-cooled in an ice bath. While stirring at 0°C, the reaction mixture was bubbled with HCl gas for 45 min. The cooling bath was removed and the reaction mixture was stirred at room temperature. for 61 hours. The reaction mixture was bubbled with nitrogen for 10 minutes to remove some HCl, cooled to 0°C and diluted with brine (200 ml), water (50 ml) and ethyl acetate (200 ml). The organic layer was separated and the aqueous layer was diluted with water (100 ml) to dissolve the remaining solids. The aqueous layer was extracted with ethyl acetate (100 ml). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated to give a brown oil. Purification by flash column chromatography using MTBE in hexanes (0% to 60%) provided the racemic product as a yellow oily solid. The racemic mixture was separated using preparative chiral HPLC (Phenomenex Lux Amylose-1 [21.2 x 250 mm, 5 µm], eluting with 95% ethanol in hexanes at a flow rate of 18 ml/min, loading about 100 mg in 2 ml ethanol) to obtain the desired the second enantiomer eluted (4.50 g, 48.8%) as a viscous yellow oil. The first enantiomer to elute had a retention time of 4.0 min. The second enantiomer eluted had a retention time of 5.3 min.

Second eluting enantiomer: 1H NMR (400 MHz, DMSO-d ₆ ) 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). LC-MS for C _w H _w BrF3NO2 (M+H)+: m/z=312.0, 314.0; found: 312.0, 314.0.

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

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 eluting) in 1.4 β-dioxane (57.2 ml) was treated with bis(pinacolato)diborone (3.49 g, 13.7 mmol) and potassium acetate (3.71 g, 37.8 mmol) and degassed with nitrogen for 5 min. The reaction mixture was treated with bis-(triphenylphosphine)palladium(II) chloride (0.482 g, 0.687 mmol), degassed for 5 min and stirred at 100°C for 2.5 h. The reaction mixture was diluted with ethyl acetate (50 ml), filtered through Celite® and further washed with ethyl acetate (100 ml). The filtrate was washed with brine, dried over sodium sulfate, filtered and concentrated to give a brown foam. Purification by flash column chromatography using MTBE in hexanes (0% to 100%) provided the desired product (3.35 g, 81.5%) as a viscous yellow foam. 1H NMR (400 MHz, DMSO-d^) 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). LC-MS for C16H22BF3NO4 (M+H)+: m/z=360.2; found: 360.1.

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

A solution of 3-bromo-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-8-amine (7.50 g, 26.7 mmol) and 3,3,3trifluoro-2-hydroxy-2-(4- methyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanamide (10.5 g, 29.4 mmol) (example 1, step 5) in 1 ,4-dioxane (133 ml) was treated with 1.0 M potassium carbonate in water (53.4 ml, 53.4 mmol), degassed with nitrogen for 5 min, treated with dichloro[1,1'-bis-(diphenylphosphino) complex )ferrocene]palladium(II) and dichloromethane (3.27 g, 4.00 mmol), degassed with nitrogen for an additional 5 minutes and stirred at 100°C for 19 hours. The reaction mixture was treated with ethyl acetate (200 ml) and brine (50 ml), filtered through Celite and washed further with ethyl acetate. The aqueous layer was separated from the filtrate and extracted with ethyl acetate (200 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated to a brown foam. Purification by flash column chromatography using MeOH in dichloromethane (0% to 10%) provided the desired product as a red-brown foam that was not completely pure. This material was repurified by flash column chromatography using MeOH in dichloromethane (0% to 15%), yielding the desired product as an orange-brown foam that was still not completely pure. This material was repurified by flash column chromatography using ethyl acetate (containing 5% MeOH) in hexanes (0% to 100%), yielding the desired product as a white foam that still contained impurities. This material was repurified by flash column chromatography using acetonitrile (containing 5% MeOH) in dichloromethane (0% to 100%) to give the desired product (4.67 g, 40.4%) as a white foam. ^1H NMR (600 MHz, DMSO-^) 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). LC-MS for C ₁₇ H ₁₄ F ₆ N ₅ O ₂ (M+H)+: m/z=434.1; found: 434.1.

Example 2. Preparation and characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2a]nazin-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]pyrazin-3-yl)-4-methylphenyl)3,3,3-trifluoro-2-hydroxypropanamide (0.050 g, 0.115 mmol) and stirred at 80°C, while adding dropwise a pre-mixed 1:2 solution of isopropyl acetate (0.676 ml)/heptane (1.34 ml). After adding 2 ml, the solid did not dissolve completely and some remained at the bottom of the vessel. After almost all the solids had dissolved, new solids formed on the walls of the vessel. After stirring at 80°C for 2 hours, more solids formed. After cooling to ambient temperature, the solids were filtered and washed with heptane. The solids were collected and dried under reduced pressure for 30 min to give 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3yl)-4-methylphenyl)-3,3 ,3-trifluoro-2-hydroxypropanamide (Form IA) (33.2 mg, 66.4%) as a white solid.

Form IA was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of Form IA is shown in FIG. 1 and peak data are given below in table. 1.

Table 1 __________________Peak XRPD data for Form IA__________________

2-theta (°) Relative intensity (%) 8.2 0.9 8.6 21.5 9.5 34.3 10.3 92.1 10.8 1.0 12.8 1.1 13.0 5.0 13.6 7.8 14.2 4.1 14.9 100

- 26 044976

DSC analysis of Form IA revealed one endothermic peak with an initial temperature of 191.9°C and a maximum at 193.2°C. The DSC thermogram is shown in Fig. 2.

TGA analysis of Form IA revealed significant mass loss above 200°C due to sample decomposition. The TGA thermogram is shown in Fig. 3.

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

Example 3. Preparation and characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2a]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[1,2a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide free base was dissolved in 1 ml of isopropyl acetate in a transparent glass bottle of 4 ml. 2 ml of heptane was added to the solution with stirring at ambient temperature. The mixture was heated at 80°C with stirring for 2 hours. The mixture was cooled to ambient temperature and stirred for 1 hour. The solid was collected by filtration and air dried to give 2-(3-(8-amino-6-(trifluoromethyl)imidazo [1,2a]nirazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (Form IIA).

Form IIA was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of Form IIA is shown in FIG. 4 and peak data are given below in table. 2.

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table 2

Peak XRPD data for Form IIA__________________

2-theta (°) Relative intensity (%) 9.1 29.8 10.0 2.2 11.1 38.6 12.6 77.6 13.5 24.8 14.1 5.6 15.4 3.6 16.1 24.0 16.9 41.4 18.0 75.7 18.4 35.0 19.0 77.5 19.7 25.5 19.9 19.5 20.1 40.9 20.5 58.2 21.0 1.1 21.4 2.1 21.6 1.3 21.9 100 23.7 29.4 23.8 39.7 25.1 22.5 25.3 33.1 25.8 33.9 26.3 13.6 26.4 3.3 27.3 35.5 28.3 13.2 29.6 15.4

DSC analysis of Form IIA revealed one endothermic peak with an initial temperature of 177.2°C and a maximum at 179.7°C. The DSC thermogram is shown in Fig. 5.

TGA analysis of Form IIA revealed significant mass loss above 200°C due to sample decomposition. The TGA thermogram is shown in Fig. 6.

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

Example 4. Preparation and characterization of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2- hydroxypropanamide, crystal form IIIA (free base)

Approximately 72 mg of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide free base was dissolved in 1 ml MeOH in a 4 ml transparent glass bottle. The solution was evaporated to a dry residue at ambient temperature. The resulting solid, which is a MeOH solvate, was dried at 60°C in vacuo overnight to give 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin3-yl)- 4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide (form IIIA).

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Form IIIA was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of Form IIIA is shown in FIG. 7 and peak data are given below in table. 3.

Table 3 __________________Peak XRPD data for Form IIIA_________________

2-theta (°) Relative intensity (%) 8.1 10.0 10.6 44.4 12.4 5.0 12.8 6.7 13.5 100 14.2 60.9 15.8 3.3 16.4 25.7 17.1 28.3 17.9 29.1 19.8 7.8 20.3 60.6 20.8 15.8 21.6 2.7 22.3 6.5 22.8 10.9 23.5 10.8 24.1 24.6 24.6 15.0 24.8 18.0 25.2 10.0 25.8 10.8 26.6 13.0 27.5 24.0 28.8 6.8 29.2 4.8 29.8 2.1

DSC analysis of Form IIIA revealed one endothermic peak with an initial temperature of 134.3°C and a maximum at 143.0°C. The DSC thermogram is shown in Fig. 8.

TGA analysis of Form IIIA revealed significant mass loss above 200°C due to sample degradation. The TGA thermogram is shown in Fig. 9.

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

Examples 5 and 6 8-Amino-X-(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)

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

To a solution of 6,8-dibromimidazo[1,2-a]pyrazine (0.50 g, 1.8 mmol) [Combi-Blocks, OR-7964] in DMF (12 ml) was added N-iodosuccinimide (0.45 g , 2.0 mmol). The reaction mixture was then heated at 60°C for 15.5 hours. The reaction mixture was concentrated in vacuo. The resulting solid was taken up in dichloromethane (DCM). The organic layer was washed successively with water and a saturated aqueous Na ₂ S ₂ O ₃ solution. The organic layer was then dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound as a light yellow solid (0.64 g, 88%). LC-MS for C ₆ H ₃ Br ₃ IN ₃ (M+H)+: calculated m/z=401.8, 403.8, 405.8; found 401.8, 403.7, 405.6.

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

A solution of 6,8-dibromo-3-iodimidazo[1,2-a]pyrazine (1.67 g, 3.57 mmol), N,N-diisopropylethylamine (1.24 ml, 7.13 mmol) and (4- methoxyphenyl)methanamine (0.512 ml, 3.92 mmol) in iPrOH (11.9 ml) was heated in a microwave reactor 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 min. The solids were collected by filtration to obtain the desired product (1.41 g, 86.1%), which was used without further purification. LC-MS for C ₁₄ H ₁₃ BrIN ₄ O (M+H)+: calculated m/z=458.9, 460.9; found 459.0, 461.0.

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

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

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

A solution of 1-(3-bromo-4-methylphenyl)ethan-1-one (1.20 g, 5.63 mmol) [Aldrich, 579734] in tetrahydrofuran (22.5 ml) at 0°C was treated with trimethyl(trifluoromethyl) silane (1.00 mL, 6.76 mmol) [Aldrich, 488712] and stirred at 0°C for 5 min. The reaction mixture was treated with 1.0 M tetra-nbutylammonium fluoride in tetrahydrofuran (0.282 ml, 0.282 mmol) at 0°C and stirred at room temperature for 1 hour. The reaction mixture was cooled to 0°C, treated with additional 1.0 M tetra -n-butylammonium fluoride in tetrahydrofuran (6.76 ml, 6.76 mmol) and stirred at room temperature for 30 minutes. The reaction mixture was diluted with ethyl acetate (100 ml) and washed with brine (2x75 ml). The organic layer was separated, dried over sodium sulfate, filtered and evaporated to obtain a crude residue. Purification by flash column chromatography using ethyl acetate in hexanes (0% to 30%) provided the desired product (1.54 g, 96.7%) as a yellow oil. LC-MS for C ₁₀ H ₉ BrF ₃ (M-OH) ⁺ : m/z=265.0, 267.0; found: 264.9, 267.0.

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Step 5. 1,1,1-Trifluoro-2-(4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan2-ol

A mixture of 2-(3-bromo-4-methylphenyl)-1,1,1-trifluoropropan-2-ol (0.252 g, 0.890 mmol), bis(pinacolato)diborone (0.294 g, 1.16 mmol) and potassium acetate ( 0.288 g, 2.94 mmol) in tetrahydrofuran (4.95 ml) was degassed with nitrogen for 5 min. The reaction mixture was treated with triphenylphosphine palladium chloride (0.025 g, 0.036 mmol), degassed with nitrogen for an additional 5 min and heated at 135°C in a microwave reactor for 20 min. The reaction mixture was diluted with ethyl acetate and filtered through a 0.5 μm cartridge, which was washed with ethyl acetate. The filtrate was washed with water and brine, dried over sodium sulfate, filtered and concentrated to give a crude residue. Purification by flash column chromatography using ether in hexanes (0% to 50%) provided the desired product (272 mg, 92.5%) as a colorless oil. LC-MS for C16H ₂ 3BF3O3 (M+H)+: m/z=331.2; found: 331.2.

Step 6. 2-(3-(8-Amino-6-bromimidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-1,1,1-trifluoropropan2-ol

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

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

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

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Step 8. 8-Amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan2-yl)phenyl)imidazo[1,2 -a]pyrazine-6-carboxamide.

Solution of methyl 8-amino-3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2a]pyrazine-6-carboxylate (step 7, 0.080 g, 0.203 mmol) in THF (3.38 ml) was treated with 1-amino-2methylpropan-2-ol (0.181 g, 2.03 mmol) and then trimethylaluminum (0.507 ml, 1.01 mmol) (2 M in toluene ) and stirred at 80°C overnight. The reaction mixture was further treated with trimethylaluminum (0.70 ml, 1.40 mmol) (2 M in toluene) and stirred at 80°C overnight. The reaction mixture was cooled to room temperature, diluted with methanol and filtered through Celite®. After washing with MeOH (2x), the filtrate was concentrated to an amber oil. Purification by silica gel chromatography (0-5% MeOH/DCM) afforded the title compound as an oily solid (26 mg, 28%), which was a mixture of enantiomers. The racemic mixture was separated using preparative chiral HPLC (Phenomenex Lux Amylose-1 [21.2 x 250 mm, 5 µm], eluting with 12% ethanol in hexanes at a flow rate of 18 ml/min, loading ~8 mg in 800 µl ethanol). The first peak eluted had a retention time of 11.9 min (Example 1; enantiomer 1). The second peak eluted had a retention time of 16.1 min (Example 2; enantiomer 2).

Example 5 (enantiomer 1). 1H NMR (400 MHz, DMSO-d ₆ ) 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). LC-MS for C ₂₁ H ₂₅ F ₃ N ₅ O ₃ (M+H)+: m/z=452.2; found: 452.1.

Example 6 (enantiomer 2). 1H NMR (600 MHz, DMSO-d ₆ ) 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). LC-MS for C ₂₁ H ₂₅ F ₃ N ₅ O ₃ (M+H)+: m/z=452.2; found: 452.2.

Example 7. Preparation and characterization of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo [1,2-a]pyrazine-6-carboxamide, crystalline form IB (free base).

8-amino-M-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1, 2-a]pyrazine-6-carboxamide (enantiomer 2 from example 6, step 8; 184 g, 408 mmol) and isopropyl acetate (950 ml). The mixture was stirred for 1 hour at 80°C, then cooled to room temperature (RT) and stirred at RT overnight. The solids were collected to give 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2hydroxypropan-2-yl)phenyl)imidazo[1 ,2-a]pyrazine-6-carboxamide (Form IB, 152 g, 82.8%).

Form IB was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of Form IB is shown in FIG. 10 and peak data are given below in table. 4.

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Table 4

Peak XRPD data for Form IB

2-theta (°) 6.2 9.6 10.4 11.4 11.6 12.0 12.4 12.6 13.9 14.4 15.1 15.6 16.0 16.7 16.9 17.4 18.3 18.9 19.3 19.9 20.2 20.7 21.0 21.3 21.9 22.3 22.9 23.2 23.8 24.1 24.4 24.8 25.0 25.5 26.0 27.1 27.3 28.0 29.1 29.7

Relative intensity (%) 31.8 0.6 8.8 12.4 6.7 15.6 4.2 3.9 6.0 10.6 0.5 100 26.8 49.1 12.6 2.9 9.0 5.4 4.3 19.4 2.6 68.4 4.0 21.9 5.2 10.4 0.6 29.4 0.5 8.9 1.3 1.5 1.1 8.2 2.3 16.8 2.2 7.9 3.0 0.4

DSC analysis of Form IB revealed one endothermic peak with an initial temperature of 172.2°C and a maximum at 174.2°C. The DSC thermogram is shown in Fig. 11. Form IB was confirmed to be an anhydrous, non-solvated crystalline form.

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Example 8. Preparation and characterization of 8-amino-K-(2-hydroxy-2-methylpropyl)-3-(2methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo [1,2-a]pyrazine-6-carboxamide, crystalline form IIB (free base).

8-amino-X-(2-hydroxy-2-methylpropyl)-3-(2-methyl-5-(1,1,1-trifluoro-2hydroxynpropan-2-yl)phenyl)imidazo[1,2 -a]pyrazine-6-carboxamide (enantiomer 2 from Example 6, Step 8; 252 mg, 0.559 mmol) and isopropyl acetate (1.25 ml), and the solids dissolved slowly. The mixture was treated with heptane (0.35 ml) until solids remained. The mixture was heated at 80°C for 30 min and stirred at RT overnight. The solids were collected to give 8-amino-X-(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%).

Form IIB was confirmed to be a crystalline solid by XRPD analysis. The XRPD pattern of Form IIB is shown in FIG. 12 and peak data are given below in table. 5.

Table 5

Peak XRPD data for Form IIB

2-theta (°) 4.3 6.2 7.4 8.6 11.3 13.3 14.7 14.9 15.3 15.5 17.0 17.2 18.1 18.8 19.6 19.8 20.1 20.8 21.4 22.4 22.7 23.5 24.1 25.1 25.8 26.2 26.5 26.9 27.3 27.9

Relative intensity (%) 59.1 0.5 100 7.6 2.1 34.1 10.5 4.1 58.1 47.3 78.5 41.1 34.9 63.6 15.1 10.0 79.7 0.6 32.3 1.5 6.9 36.0 6.8 6.3 28.3 18.0 13.9 1.0 19.5 14.6

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DSC analysis of Form IIB revealed one endothermic peak with an initial temperature of 161.7°C and a maximum at 165.4°C. The DSC thermogram is shown in Fig. 8. Form IIB was confirmed to be an anhydrous, non-solvated crystalline form.

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

Step 1. 1-(4-(Methyl-d ₃ )phenyl)ethan-1-one

A solution of (4-acetylphenyl)boronic acid (1.00 g, 6.10 mmol) [Aldrich, 470821], bis-(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II) (0.108 g, 0.152 mmol) and Cesium fluoride (3.24 g, 21.4 mmol) in DMF (10.2 ml) and water (2.03 ml) was degassed with nitrogen for 10 min, treated with iodomethane-d ₃ (1.44 ml, 23.2 mmol), and stirred at 45°C overnight. The reaction mixture was cooled to room temperature. and diluted with water and ethyl acetate. The aqueous layer was separated and extracted with ethyl acetate (2x). The combined organic extracts were washed with water and brine, dried over magnesium sulfate, filtered and concentrated (60-70 Torr, 25°C bath) to give the desired product (546 mg, 65.3%) as a yellow oil, which was used without further cleaning. 1H NMR (400 MHz, CDCl ₃ ) δ 7.86 (d, J= 8.0 Hz, 2H), 7.26 (d, J= 8.0 Hz, 2H), 2.58 (s, 3H) . LC-MS for C ₉ H ₈ D ₃ O (M+H) ⁺ : m/z=138.1; found: 138.1.

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

A suspension of aluminum chloride (13.6 g, 102 mmol) in dichloromethane (24 ml) was treated dropwise through a syringe with 1-(4-(methyl-d ₃ )phenyl)ethan-1-one (6.35 g) over 5 min. , 46.3 mmol). The residual material in the syringe was washed with dichloromethane (7.0 ml) and added dropwise to the reaction mixture. After the initial exotherm, the reaction mixture was allowed to cool to room temperature. for 3 min, stirred at 35°C for 5 min, and treated dropwise with bromine (2.38 ml, 46.3 mmol) for 5 min. The reaction mixture was stirred for 25 minutes and then slowly added to a mixture of dichloromethane (50 ml), 1N. HCl (100 ml) and ice. The residual reaction mixture was washed in dichloromethane/HCl/ice with additional dichloromethane. The mixture was heated to room temperature (RT) and the layers were separated. The aqueous layer was extracted with dichloromethane (2x75 ml). The combined organic layers were washed with saturated sodium hydrogencarbonate solution and brine. The sodium bicarbonate and saline washes contained product and were combined, acidified with 1 M HCl and extracted with dichloromethane (2x50 ml). All organic layers were combined, dried over magnesium sulfate, filtered and concentrated to give a yellow oil. Purification by flash column chromatography using ethyl acetate in hexanes (0% to 15%) provided the desired product (9.08 g, 90.8%) as a light yellow solid. 1H NMR (400 MHz, CDCl ₃ ) δ 8.11 (d, J=1.8 Hz, 1H), 7.79 (dd, J=7.9, 1.8 Hz, 1H), 7.32 ( d, J=7.9 Hz, 1H), 2.57 (s, 3H). LC-MS for C ₉ H ₇ D ₃ BrO (M+H)+: m/z=216.0, 218.0; found: 216.0, 218.0.

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

A solution of 1-(3-bromo-4-(methyl ₃ )phenyl)ethan-1-one (9.08 g, 42.0 mmol) in tetrahydrofuran (168 ml) at 0°C was treated with trimethyl(trifluoromethyl)silane (8.07 ml, 58.8 mmol) [Aldrich, 488712] and stirred at 0°C for 5 min. 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 (3x50 ml). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and concentrated to give a crude residue. Purification by flash column chromatography using ethyl acetate in hexanes (0% to 20%) gave the desired product (13.3 g, 111%) as a yellow oil. 1H NMR (400 MHz, CDCl ₃ ) 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).

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

A suspension of bis(pinacolato)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-d ₃ )phenyl)-1,1,1-trifluoropropan-2-ol (13.3 g, 41.8 mmol). Residual 2-(3-bromo-4-(methyl ₃ )phenyl)-1,1,1-trifluoropropan-2-ol was washed with dioxane (106 ml) and added to the reaction mixture, which was degassed with nitrogen for 10 minutes. The reaction mixture was treated with bis-(triphenylphosphine)palladium(II) chloride (1.16 g, 1.67 mmol), degassed with nitrogen for a further 10 min and stirred at 100°C overnight. The reaction mixture was cooled to room temperature, degassed with nitrogen for 5 min, further treated with bis(triphenylphosphine) palladium(II) chloride (1.16 g, 1.67 mmol), degassed with nitrogen for an additional 5 min and stirred at 100 °C for 4 hours. The reaction mixture was filtered through Celite® and washed with THF and ethyl acetate. The filtrate was washed with water/saline 1:1 (300 ml). The aqueous layer was re-extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered and concentrated to a brown oil. Purification by flash column chromatography using MTBE in hexanes (0% to 20%) provided the desired product (14.4 g, 84.7%) as a pale yellow oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.93 (d, J=2.3 Hz, 1H), 7.557.45 (m, 1H), 7.19 (d, J=8.1 Hz, 1H ), 2.43 (br s, 1H), 1.77 (s, 3H), 1.34 (s, 12H). LC-MS for C ₁₆ H ₂₀ D ₃ BF ₃ O ₃ (M+H)+: m/z=334.2; found: 334.3.

Step 5. 2-(3-(8-Amino-6-bromimidazo[1,2-v]pyrazin-3-yl)-4-(methyl ₃ )phenyl)-1,1,1-trifluoropropane-2 -ol (racemic mixture)

A solution of 1,1,1-trifluoro-2-(4-(methyl ₃ )-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan- 2-ol (14.5 g, 35.6 mmol) in dioxane (178 ml) was treated with 6-bromo-3-iodumidazo[1,2-a]pyrazin-8amine (12.1 g, 35.6 mmol), degassed with nitrogen for 5 min, treated with a complex of dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) and dichloromethane (5.81 g, 7.11 mmol) and degassed with nitrogen for another 5 min.

The reaction mixture was treated with 1.0 M potassium carbonate in water (107 ml, 107 mmol), degassed with nitrogen for 5 min and stirred at 80°C overnight. The reaction mixture was cooled to room temperature. and filtered through Celite®. Celite® was washed with ethyl acetate and water. Filtrate diluted

- 36 044976 was poured with water (150 ml) and extracted with ethyl acetate (3x100 ml). The combined organic layers were dried over magnesium sulfate, filtered and concentrated to a dark oil. Purification by flash column chromatography using methanol in dichloromethane (0% to 5%) and repurification by flash column chromatography using ethyl acetate in hexanes (0% to 100%) provided the desired product (13.8 g, 92.8%). LC-MS for C ₁₆ H ₁₂ D ₃ BrF ₃ N ₄ O (M+H)+: m/z=418.1, 420.1; found: 418.0, 420.0.

Step 6. Second eluted enantiomer 2-(3-(8-αmino-6-bromo-midazo[1,2-a]pyrazin-3-ul)-4(methyl-d ₃ )phenyl)-1,1,1-trifluoropropane -2-ola

The racemic mixture of 2-(3-(8-amino-6-bromimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d ₃ )phenyl)-1,1,1trifluoropropan-2-ol was separated by preparative chiral HPLC (Phenomenex Lux Amylose1 [21.2x250 mm, 5 µm], eluting with 20% ethanol in hexanes at a flow rate of 20 ml/min, loading ~200 mg in 4 ml ethanol). The first peak to elute had a retention time of 9.6 min. The second eluted peak had a retention time of 14.6 min.

Peak 2: 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). LC-MS for C ₁₆ H ₁₂ D ₃ BrF ₃ N ₄ O (M+H)+: m/z=418.1, 420.1; found: 418.0, 420.0.

Step 7. Methyl 8-amino-3-(2-(methyl-d ₃ )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]nirazine -6-carboxylate (the only enantiomer obtained)

Solution of 2-(3-(8-amino-6-bromimidazo[1,2-a]pyrazin-3-yl)-4-(methyl-d ₃ )phenyl)-1,1,1-trifluoropropan-2-ol (peak 2 of step 6, 4.95 g, 48.9 mmol) in methanol (163 ml) and DMF (40.7 ml) was treated with triethylamine (6.81 ml, 48.9 mmol), and degassed with nitrogen for 5 min. The reaction mixture was treated with Pd(dppf) ₂ CH ₂ Cl ₂ (0.998 g, 1.22 mmol), and degassed with nitrogen for an additional 5 minutes, saturated with CO by bubbling the gas under the surface of the reaction mixture for 3 minutes, and heated at 60° C overnight. The reaction mixture was concentrated and the resulting oil was diluted with ethyl acetate and water. The aqueous layer was separated and re-extracted with ethyl acetate (3x). The combined organic layers were washed with water, saturated ammonium chloride and brine, dried over magnesium sulfate, filtered and concentrated to give a brown oil. Purification by flash column chromatography using methanol in dichloromethane (0% to 5%) provided the desired product (4.49 g, 92.4%) as an orange oily solid. LC-MS for C ₁₈ H ₁₅ D ₃ F ₃ N ₄ O ₃ (M+H)+: m/z=398.1; found: 398.3.

Step 8. 8-Amino-3-(2-(methyl-d ₃ )-5 -(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2a]pyrazin-6- carboxylic acid (the only enantiomer obtained)

A solution of methyl 8-amino-3 -(2-(methyl-d ₃ )-5 -(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2-a]nirazin-6 -carboxylate (4.49 g, 11.3 mmol) (the only enantiomer obtained in step 7) in methanol (113 ml) was treated with 1.0 M sodium hydroxide (56.5 ml, 56.5 mmol) and stirred at room temperature. The reaction mixture was concentrated to remove methanol, diluted with water (50 ml), and extracted with ethyl acetate (50 ml, then 20 ml). The combined ethyl acetate layers were extracted with additional 1.0 M sodium hydroxide (3x20 ml). pH level of combined basics

- 37 044976 aqueous layers were brought to ~5 with citric acid (7.6 g). The aqueous layer was extracted with dichloromethane (2x150 ml). The aqueous layer was diluted with brine and extracted with ethyl acetate (150 ml). The combined organic layers were concentrated to give the desired product (4.06 g, 93.8%) as a tan solid, which was used without further purification. ¹ H NMR (400 MHz, DMSO-d ₆ ) 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 (lat s, 2H), 6.66 (s, 1H), 1.71 (s, 3H). LC-MS for C ₁₇ H ₁₃ D ₃ F ₃ N ₄ O ₃ (M+H)+: m/z=384.1; found: 384.2.

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

Solution of 8-amino-3-(2-(methyl ₃ )-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)imidazo[1,2a]pyrazine-6-carboxylic acid (4.06 g, 10.6 mmol) (sole enantiomer obtained in step 8) in DMF (106 ml) was treated with 1-amino-2-methylpropan-2-ol (1.44 g, 16.2 mmol) [ Ark Pharm, AK37803] and HATU (6.16 g, 16.2 mmol), stirred for 15 min, treated with triethylamine (4.43 ml, 31.8 mmol) and stirred at room temperature. for 3.5 hours. The reaction mixture was diluted with water (500 ml) and brine (100 ml) and extracted with ethyl acetate (3x150 ml). The combined organic layers were washed with saturated ammonium chloride (150 ml), 11% sodium carbonate (150 ml) and brine (100 ml), dried over magnesium sulfate, filtered and concentrated to give an amber oil. Purification by flash column chromatography using methanol in dichloromethane (0% to 5%) provided the desired product (4.28 g, 89.0%) as a foam. ¹ H NMR (600 MHz, DMSO-d ₆ ) 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). LC-MS for C ₂₁ H ₂₂ D ₃ F ₃ N ₅ O ₃ (M+H)+: m/z=455.2; found: 455.2.

Example 10. Preparation and characterization of 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2(methyl-d ₃ )-5-(1,1,1-trifluoro-2-hydroxypropane-2 -yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, crystalline form IC (free base).

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

Based on the results of XRPD analysis, the IC form was confirmed to be a crystalline solid. The XRPD pattern of the IC shape is shown in FIG. 14 and peak data are given below in table. 6.

Table 6

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DSC analysis of the IC form revealed one endothermic peak with an initial temperature of 173.4°C and a maximum at 179.0°C. The DSC thermogram is shown in Fig. 15. Form IC was confirmed to be an anhydrous, non-solvated crystalline form.

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

2-(3-(8-Amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)3,3,3-trifluoro-2-hydroxypropanamide free base (98, 81 mg) was dissolved in 2.5 ml of methanol in a 4 ml transparent glass vial. 42.4 μL of 6 M HBr aqueous solution (1.2 eq) was added to the solution and mixed well. The solution was evaporated at room temperature to obtain HBr salt crystals.

Example 12 Characterization of a single crystal of the hydrobromic acid (HBr) salt 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3, 3,3-trifluoro-2hydroxypropanamide.

Crystal data: C35 H32 Br2 F12 N10 O5, from methanol, colorless, irregular plate, -0.450x0.210x0.060 mm, monoclinic, C2, a=20.055(7) A, b=10.115(4) A, c= 21.363(8) ±, beta=94.953(7), vol. = 4318(3) A ³ , Z=4, T=-40°C, mass in accordance with the formula=1060.52, density=1.631 g/cm ³ , µm(Mo) = 1.98 mm ^-1 .

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Data collection. Data acquisition was performed using a Bruker SMART APEX-II CCD system, MoKalpha radiation, standard focusing tube, anode power = 50 kV x 30 mA, die to wafer distance = 5.0 cm, 512 x 512 pixels/frame, beam center = (259.19 ,253.13), total number of frames=2635, oscillation/frame=0.50°, shutter speed/frame=40.1 s/frame, SAINT integration, hkl min/max = (-26.26,-12.13,- 27.27), data input in shelx=38968, number of unique data=9756, dvate range=4.51 to 55.43°, completeness for dva-theta 55.43=99.60%, R(int-xl ) = 0.0672, applying the SADABS correction.

Solution and clarification.

The crystal structure was determined using XS(Shelxtl), and refinement was performed using the shelxtl software package. Refinement was carried out using the full-matrix least squares method on F ² , scattering coefficients from ref. table Volume C, Table. 4.2.6.8 and 6.1.1.4, number of data=9756, number of restrictions=1, number of parameters=584, data/parameter ratio=16.71, goodness-of-fit criterion for F2=1.14, indices R [I>4 sigma(I )] R1=0.0648, wR2=0.1560, R indices (all data) R1=0.1004, wR2=0.1719, maximum difference between peak and hole=1.795 and -0.642 e/A ³ , improved Flack parameter=0.038( 6). All hydrogen atoms are refined in the rider model. In table Figure 7 shows the atomic coordinates (x10 ⁴ ) and equivalent isotropic shear parameters (A ² x10 ³ ). u(eq.) is defined as one third of the projection of the orthogonalized tensor Uij. In table Figure 8 shows the values of bond length [A] and angles [deg]. In table Figure 9 shows the parameters of the anisotropic displacement (A ² x10 ³ ).

Results.

This analysis confirmed the structure of the hydrobromic acid salt 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro- 2-hydroxypropanamide. The asymmetric group contains two molecules of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide , two bromides for charge balancing and one molecule of methanol solvent, as shown in Fig. 16A-16B. The enantiomeric parameter was based on Flack's parameter, which was refined to 0.038(6). This study determined the absolute configuration of the C15=S- and C35=S- chiral centers.

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Table 7 x y z i(eq)

Вг(1) 5582(1) 9450(1) 4009(1) 42(1) Вг(2) 5862(1) 10538(1) 1429(1) 55(1) F(l) 6628(3) 14263( 6) 1224(3) 55(2) F(2) 6580(3) 15776(7) 1906(3) 69(2) F(3) 5777(3) 15548(8) 1169(3) 76(2) F(4) 2936(3) 16054(7) 74(2) 64(2) F(5) 2172(3) 16617(6) 663(3) 53(2) F(6) 2941(3) 17995( 6) 485(3) 64(2) F(21) -154(3) 10814(6) 4245(3) 56(2) F(22) -36(4) 9202(6) 3634(4) 77( 2) F(23) 718(3) 9617(9) 4368(4) 95(3) F(24) 2863(3) 8853(7) 5238(2) 50(2) F(25) 2167(2) 8776(5) 4410(3) 42(1) F(26) 2742(3) 7056(5) 4708(3) 49(1) 0(1) 3939(3) 16429(6) 1010(3) 37( 1) 0(2) 3666(3) 17493(7) 2068(3) 43(2) 0(21) 3946(3) 8374(6) 4603(3) 32(1) 0(22) 2767(3) 7895(7) 3331(3) 36(2) 0(40) 4500(4) 14342(10) 397(4) 75(2) N(l) 4949(3) 11437(8) 3026(3) 36( 2) N(2) 6280(3) 13465(8) 2462(4) 36(2) N(3) 4935(3) 12867(7) 2259(3) 28(2) N(4) 6392(4) 11972(9) 3289(4) 45(2) N(5) 2564(4) 17096(8) 1956(4) 39(2) N(21) 1533(4) 13615(8) 2454(4) 33( 2) N(22) 219(3) 11535(8) 3008(4) 39(2) N(23) 1563(3) 12205(7) 3219(3) 31(2) N(24) 98(4) 12981(9) 2172(4) 50(2) N(25) 3891(3) 7901(8) 3415(3) 39(2) C(l) 4279(4) 12394(8) 2289(4) 28( 2) C(2) 4303(4) 11526(9) 2766(4) 32(2)

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C(3) 5320(4) 12256(9) 2717(4) 31(2)

C(4) 6017(4) 12567(10) 2820(4) 35(2)

C(5) 5853(4) 13980(9) 1980(4) 33(2)

C(6) 5199(4) 13700(9) 1858(4) 34(2)

C(7) 6210(5) 14906(11) 1569(5) 49(3)

C(8) 3706(4) 12918(9) 1867(4) 31(2)

C(9) 3210(4) 12062(10) 1606(4) 35(2)

C(10) 2685(4) 12628(10) 1211(4) 39(2)

C(ll) 2677(4) 13941(10) 1064(4) 34(2)

C(12) 3181(4) 14780(9) 1310(4) 29(2)

C(13) 3678(4) 14244(8) 1725(3) 26(2)

C(14) 3214(5) 10601(11) 1716(6) 54(3)

C(15) 3278(4) 16231(9) 1162(4) 28(2)

C(16) 2819(5) 16748(10) 589(4) 36(2)

C(17) 3181(4) 17041(9) 1765(4) 29(2)

C(21) 2216(4) 12663(9) 3209(4) 31(2)

C(22) 2176(4) 13579(10) 2730(4) 36(2)

C(23) 1158(4) 12776(9) 2745(4) 32(2)

C(24) 462(4) 12442(10) 2644(4) 34(2)

C(25) 628(4) 11023(9) 3491(4) 35(2)

C(26) 1280(4) 11327(8) 3631(4) 33(2)

C(27) 287(4) 10187(10) 3932(5) 42(2)

C(28) 2774(4) 12198(8) 3629(4) 27(2)

C(29) 3264(4) 13027(8) 3916(4) 30(2)

C(30) 3753(5) 12484(9) 4315(4) 35(2)

C(31) 3802(4) 11136(9) 4439(4) 32(2)

C(32) 3317(4) 10301(8) 4149(4) 22(2)

C(33) 2819(4) 10830(8) 3738(4) 26(2)

C(34) 3239(5) 14511(10) 3811(5) 45(2)

C(35) 3344(4) 8814(9) 4276(4) 29(2)

C(36) 2773(4) 8377(9) 4650(4) 34(2)

C(37) 3309(4) 8112(9) 3631(4) 27(2)

C(41) 4168(10) 13090(30) 222(9) 147(10)

Table 8

F(l)-C(7) 1.332(12)

F(2)-C(7) 1.323(12)

F(3)-C(7) 1.334(12)

F(4)-C(16) 1.343(11)

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F(5)-C(16) 1.327(11)

F(6)-C(16) 1.307(11)

F(21)-C(27) 1.315(11)

F(22)-C(27) 1.322(12)

F(23)-C(27) 1.345(12)

F(24)-C(36) 1.342(10)

F(25)-C(36) 1.341(10)

F(26)-C(36) 1.344(10)

O(l)-C(15) 1.405(10)

O(2)-C(17) 1.211(10)

O(21)-C(35) 1.414(10)

O(22)-C(37) 1.232(9)

O(40)-C(41) 1.46(2)

N(l)-C(3) 1.327(11)

N(l)-C(2) 1.368(10)

N(2)-C(4) 1.326(12)

N(2)-C(5) 1.382(11)

N(3)-C(6) 1.343(11)

N(3)-C(3) 1.342(11)

N(3)-C(l) 1.407(10)

N(4)-C(4) 1.341(11)

N(5)-C(17) 1.338(11)

N(21)-C(23) 1.323(12)

N(21)-C(22) 1.372(11)

N(22)-C(24) 1.323(12)

N(22)-C(25) 1.364(12)

N(23)-C(23) 1.369(11)

N(23)-C(21) 1.392(10)

N(23)-C(26) 1.404(11)

N(24)-C(24) 1.311(11)

N(25)-C(37) 1.310(10)

C(l)-C(2) 1.344(12)

C(l)-C(8) 1.494(11)

C(3)-C(4) 1.432(12)

C(5)-C(6) 1.345(12)

C(5)-C(7) 1.506(13)

C(8)-C(13) 1.376(12)

C(8)-C(9) 1.399(12)

C(9)-C(10) 1.413(13)

C(9)-C(14) 1.497(15)

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C(1O)-C(11) 1.365(14)

C(ll)-C(12) 1.388(12)

C(12)-C(13) 1.386(11)

C(12)-C(15) 1.518(12)

C(15)-C(17) 1.552(12)

C(15)-C(16) 1.556(11)

C(21)-C(22) 1.377(12)

C(21)-C(28) 1.450(11)

C(23)-C(24) 1.435(12)

C(25)-C(26) 1.351(12)

C(25)-C(27) 1.478(13)

C(28)-C(29) 1.394(12)

C(28)-C(33) 1.404(12)

C(29)-C(30) 1.359(12)

C(29)-C(34) 1.517(13)

C(30)-C(31) 1.391(13)

C(31)-C(32) 1.393(11)

C(32)-C(33) 1.380(11)

C(32)-C(35) 1.529(12)

C(35)-C(36) 1.518(11)

C(35)-C(37) 1.546(11)

C(3)-N(l)-C(2) 107.9(7)

C(4)-N(2)-C(5) 115.9(7)

C(6)-N(3)-C(3) 121.4(7)

C(6)-N(3)-C(l) 131.5(7)

C(3)-N(3)-C(l) 107.0(7)

C(23)-N(21)-C(22) 109.4(8)

C(24)-N(22)-C(25) 118.7(7)

C(23)-N(23)-C(21) 110.3(7)

C(23)-N(23)-C(26) 119.0(7)

C(21)-N(23)-C(26) 130.7(7)

C(2)-C(l)-N(3) 106.5(7)

C(2)-C(l)-C(8) 131.6(7)

N(3)-C(l)-C(8) 121.8(7)

C(l)-C(2)-N(l) 108.8(7)

N(l)-C(3)-N(3) 109.8(7)

N(l)-C(3)-C(4) 129.9(8)

N(3)-C(3)-C(4) 120.2(8)

N(2)-C(4)-N(4) 120.9(8)

N(2)-C(4)-C(3) 119.5(8)

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N(4)-C(4)-C(3) 119.6(8) C(6)-C(5)-N(2) 126.5(8) C(6)-C(5)-C(7) 121.6(8) N(2)-C(5)-C(7) 111.9(8) N(3)-C(6)-C(5) 116.1(8) F(2)-C(7)- F(l) 106.0(8) F(2)-C(7)-F(3) 109.1(9) F(l)-C(7)-F(3) 106.8(9) F(2)-C (7)-C(5) 111.7(9) F(l)-C(7)-C(5) 111.8(9) F(3)-C(7)-C(5) 111.1(8) C( 13)-C(8)-C(9) 120.0(8) C(13)-C(8)-C(l) 119.6(7) C(9)-C(8)-C(l) 120.3( 8) C(8)-C(9)-C(10) 117.1(9) C(8)-C(9)-C(14) 123.6(8) C(10)-C(9)-C( 14) 119.2(8) C(ll)-C(10)-C(9) 121.7(9) C(10)-C(ll)-C(12) 121.0(8) C(13)-C(12 )-C(ll) 117.6(8) C(13)-C(12)-C(15) 114.4(7) C(ll)-C(12)-C(15) 128.0(8) C(8) -C(13)-C(12) 122.5(7) O(l)-C(15)-C(12) 109.1(7) O(l)-C(15)-C(17) 107.7(7)

C(12)-C(15)-C(17) 108.1(7) O(l)-C(15)-C(16) 106.0(7) C(12)-C(15)-C(16) 114.2(7) C(17)-C(15)-C(16) 111.5(7) F(6)-C(16)-F(5) 108.3(7) F(6)-C(16)- F(4) 108.5(8) F(5)-C(16)-F(4) 106.6(8) F(6)-C(16)-C(15) 110.6(8) F(5)-C (16)-C(15) 113.0(7) F(4)-C(16)-C(15) 109.7(7) O(2)-C(17)-N(5) 122.8(8) O( 2)-C(17)-C(15) 119.4(8) N(5)-C(17)-C(15) 117.5(8) C(22)-C(21)-N(23) 103.9( 7) C(22)-C(21)-C(28) 132.1(8)

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N(23)-C(21)-C(28) 124.0(7)

N(21)-C(22)-C(21) 109.2(8)

N(21)-C(23)-N(23) 107.0(7)

N(21)-C(23)-C(24) 131.7(8)

N(23)-C(23)-C(24) 121.3(8)

N(24)-C(24)-N(22) 122.1(8)

N(24)-C(24)-C(23) 119.2(9)

N(22)-C(24)-C(23) 118.5(8)

C(26)-C(25)-N(22) 126.0(8)

C(26)-C(25)-C(27) 118.9(9)

N(22)-C(25)-C(27) 114.7(8)

C(25)-C(26)-N(23) 116.2(8)

F(21)-C(27)-F(22) 106.6(7)

F(21)-C(27)-F(23) 106.0(9)

F(22)-C(27)-F(23) 105.7(9)

F(21)-C(27)-C(25) 114.6(8)

F(22)-C(27)-C(25) 111.0(9)

F(23)-C(27)-C(25) 112.4(7)

C(29)-C(28)-C(33) 119.1(7)

C(29)-C(28)-C(21) 123.6(8)

C(33)-C(28)-C(21) 117.3(7)

C(30)-C(29)-C(28) 118.4(8)

C(30)-C(29)-C(34) 120.3(8)

C(28)-C(29)-C(34) 121.2(8)

C(29)-C(30)-C(31) 123.5(8)

C(30)-C(31)-C(32) 118.3(8)

C(33)-C(32)-C(31) 119.2(8)

C(33)-C(32)-C(35) 120.5(7)

C(31)-C(32)-C(35) 120.3(7)

C(32)-C(33)-C(28) 121.4(8)

O(21)-C(35)-C(36) 107.4(7)

O(21)-C(35)-C(32) 114.4(7)

C(36)-C(35)-C(32) 111.2(7)

O(21)-C(35)-C(37) 105.6(7)

C(36)-C(35)-C(37) 110.9(7)

C(32)-C(35)-C(37) 107.3(7)

F(25)-C(36)-F(24) 107.1(7)

F(25)-C(36)-F(26) 106.6(7)

F(24)-C(36)-F(26) 105.9(7)

F(25)-C(36)-C(35) 114.3(7)

F(24)-C(36)-C(35) 110.1(7)

F(26)-C(36)-C(35) 112.4(7)

O(22)-C(37)-N(25) 124.1(7)

O(22)-C(37)-C(35) 121.1(7)

N(25)-C(37)-C(35) 114.5(7)

- 46 044976

Table 9

Ull U22 U33 U23 U13 U12

Br(l) 47(1) 38(1) 38(1) 8(1) -8(1) 5(1) Br(2) 62(1) 48(1) 52(1) 12(1) - 1(1) 10(1) F(l) 56(4) 48(4) 66(4) -1(3) 28(3) -3(3) F(2) 77(4) 53(4) 79(4) -12(4) 25(3) -43(4) F(3) 50(4) 68(5) 110(5) 59(5) 15(4) 8(4) F(4) 82(5) 80(5) 29(3) 2(3) -6(3) 13(4) F(5) 42(3) 56(4) 58(3) 8(3) -17(3) 10(3) F(6) 84(5) 43(4) 62(4) 21(3) -6(3) 10(3) F(21) 53(4) 53(4) 68(4) 8 (3) 28(3) 10(3) F(22) 95(5) 38(4) 107(5) -13(4) 52(4) -32(4) F(23) 43(4) 103 (7) 143(7) 85(6) 21(4) 6(4) F(24) 46(3) 68(4) 40(3) -9(3) 17(2) -8(3) F (25) 20(2) 46(3) 61(3) 8(3) 7(2) 0(2) F(26) 55(3) 25(3) 70(4) 8(3) 17(3 ) -7(2) 0(1) 31(3) 43(4) 37(3) 9(3) 10(3) 5(3) 0(2) 37(4) 51(5) 40(3) -5(3) 1(3) -7(3) 0(21) 30(3) 34(4) 31(3) 0(3) -8(2) 1(3) 0(22) 19(3 ) 45(4) 44(3) -13(3) -3(2) -2(3) 0(40) 72(5) 83(7) 72(5) 6(5) 14(4) -2 (5) N(l) 27(4) 34(5) 45(4) 12(4) -4(3) -3(3) N(2) 20(3) 39(5) 49(5) 4 (4) -2(3) 0(3) N(3) 19(3) 31(4) 35(4) 6(3) -2(3) 0(3) N(4) 28(4) 56 (6) 50(5) 16(4) -7(3) -7(4) N(5) 26(4) 46(5) 44(4) -12(4) 2(3) 3(3) N(21) 27(4) 35(5) 37(4) 13(4) 2(3) 6(3) N(22) 20(4) 36(5) 62(5) 2(4) 6( 3) 1(3) N(23) 21(3) 28(4) 45(4) 7(3) 10(3) 5(3) N(24) 25(4) 67(7) 58(5) 15(5) -4(4) -10(4) N(25) 23(4) 59(6) 35(4) -18(4) -5(3) 4(3) C(l) 17( 4) 24(4) 41(5) 8(4) -1(3) 1(3)

- 47 044976

C(2) 18(4) 33(5) 44(5) 3(4) -2(3) -2(3)

C(3) 22(4) 29(5) 44(5) 8(4) 2(4) -4(4)

C(4) 27(5) 34(5) 44(5) 3(4) 1(4) 2(4)

C(5) 26(4) 29(5) 45(5) 8(4) 5(4) 2(4)

C(6) 27(4) 33(5) 41(5) 6(4) 3(4) 1(4)

C(7) 36(5) 41(6) 70(7) 14(5) 12(5) -4(5)

C(8) 26(4) 35(5) 31(4) -3(4) 0(3) 5(4)

C(9) 25(4) 34(6) 47(5) -6(4) -2(4) -4(4)

C(10) 30(5) 43(6) 44(5) -1(5) -4(4) -5(4)

C(ll) 33(5) 39(5) 31(4) 3(4) -7(4) 0(4)

C(12) 24(4) 30(5) 34(4) -4(4) 6(3) -6(3)

C(13) 29(4) 22(5) 28(4) -1(3) -2(3) -2(3)

C(14) 41(5) 33(6) 85(8) 9(6) -9(5) -5(5)

C(15) 31(4) 26(5) 27(4) 3(3) -4(3) 5(3)

C(16) 42(5) 35(5) 30(4) 10(4) -8(4) 11(4)

C(17) 28(4) 29(5) 30(4) 5(4) -2(3) 6(4)

C(21) 22(4) 29(5) 42(5) 4(4) 7(4) 0(3)

C(22) 25(4) 34(5) 48(5) 7(4) 3(4) 5(4)

C(23) 25(4) 31(5) 42(5) 1(4) 4(4) 9(4)

C(24) 24(4) 38(5) 40(5) 2(4) 0(4) 4(4)

C(25) 29(5) 22(5) 54(6) 3(4) 9(4) 1(4)

C(26) 34(5) 17(5) 48(5) 5(4) 10(4) 4(3)

C(27) 28(5) 30(6) 69(7) 2(5) 8(5) -2(4)

C(28) 18(4) 27(5) 37(4) 4(4) 6(3) 3(3)

C(29) 30(4) 20(5) 39(5) 0(4) 4(4) -5(3)

C(30) 35(5) 28(5) 41(5) -7(4) 2(4) -8(4)

C(31) 32(4) 26(5) 37(5) -2(4) -2(4) -3(4)

C(32) 18(4) 23(5) 26(4) -4(3) 4(3) -1(3)

C(33) 22(4) 25(5) 32(4) 1(3) 3(3) -3(3)

C(34) 46(5) 20(4) 67(6) -3(5) 3(4) -10(5)

C(35) 21(4) 33(5) 31(4) -3(4) 0(3) -3(3)

C(36) 36(5) 32(5) 35(5) -2(4) 10(4) -1(4)

C(37) 21(4) 28(5) 30(4) -2(3) -2(3) -1(3)

C(41) 105(14) 210(30) 140(16) -79(17) 57(12) -77(16)

Example A: THP-1 RPS6 ELISA.

To measure phosphorylated ribosomal protein S6 (RPS6) in cell lysates, THP-1 (human acute monocytic leukemia) cells were purchased from ATCC (Manassas, VA) and maintained in RPMI with 10% fetal bovine serum (FBS) (Gibco/Life Technologies , Carlsbad, California, USA). For this assay, THP-1 cells were maintained in serum-free RPMI overnight, then seeded in RPMI (2 × 10 ⁵ cells/90 μl well) in 96-well flat-bottom treated tissue culture plates (Corning, Corning, NY, USA) in the presence or absence of a range of concentrations of the test compounds. Coated plates were incubated for 2 h at 37°C, 5% CO2, then treated with or without 10 nM MCP-1 (MYBioSource, San Diego, CA) for 15 min at 37°C, 5% CO2. The plates were centrifuged at 1600 rpm and the supernatants were removed. Cells were lysed in lysis buffer (Cell Signaling, Danvers, MA) with protease inhibitor (Calbiochem/EMD, Germany), PMSF (Sigma, St. Louis, MO), HALTS (Thermo Fisher, Rockford, IL) for 30 minutes on regular ice. Cell lysates were frozen at −80°C prior to analysis. Lysates were tested using a human/mouse/rat phospho-RPS6 ELISA (R&D Systems, Inc., Minnesota, MN). Plate measurements were performed using a microplate spectrophotometer (SpectraMax M5 - Molecular Devices, LLC Sunnyvale, California) set at 450 nm with a wavelength correction of 540. Determination of IC ₅ o was performed by fitting the percentage of inhibitor inhibition to the logarithm of inhibitor concentration using GraphPad Prism 5.0 software.

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Example B. Scintillation analysis of ΡΙ3Κ-γ approach.

Materials.

[γ- ^33Ρ ]ATP (10 mCi/ml) and YSi SPA wheat germ agglutinin (WGA) scintillation beads were purchased from Perkin-Elmer (Waltham, MA). Lipid kinase substrate, D-myo-phosphatidylinositol-4,5-bisphosphate (PtdIns (4,5)P2)D (+)-sn-1,2-di-O-octanoylglyceryl, 3-Ophospho-linked (PIP2), CAS 204858-53-7, purchased from Echelon Biosciences (Salt Lake City, UT). Recombinant human PI3Ky protein (p110Y) was purchased from Life technology (Grand Island, NY). ATP, MgCl ₂ , DTT, EDTA, MOPS, and CHAPS were purchased from SigmaAldrich (St. Louis, MO).

Kinase reactions were performed in Greiner Bio-one 384-well white polystyrene plates from Thermo Fisher Scientific in a final volume of 25 μl. The inhibitors were first serially diluted in DMSO and then added to the wells of the plate before adding other reaction components. The final concentration of DMSO in the assay was 2%. The PI3Ky assay was performed at room temperature in 20 mM mOpC, pH 6.7, 10 mM MgCl ₂ , 5 mM DTT and CHAPS 0.03%. Reactions were initiated by the addition of ATP, the final reaction mixture consisting of 20 µM PIP2, 2 µM ATP, 0.5 µCi [y- ^33P ]ATP, 13 nM PI3Ky. The reaction mixtures were incubated for 120 min and the reaction was stopped by adding 40 μl of SPA beads suspended in quenching buffer: 163 mM potassium phosphate, pH 7.8, 20% glycerol, 25 mM EDTA. The final concentration of SPA beads was 1.0 mg/mL. After sealing the plates, the plates were shaken overnight at room temperature and centrifuged at 1500 rpm for 10 min. The radioactivity of the product was determined by scintillation counting on a Topcount instrument (Perkin - Elmer). Determination of IC ₅₀ was performed by fitting a curve of the percentage of control solvent activity versus the logarithm of inhibitor concentration using GraphPad Prism 6.0 software.

Example C: PI3Kδ Proximity Scintillation Assay.

Materials.

[Y- ^33P ]ATP (10 mCi/ml) and YSi SPA wheat germ agglutinin (WGA) scintillation beads were purchased from Perkin-Elmer (Waltham, MA). Lipid kinase substrate, D-myo-phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2)D(+)-sn-1,2-di-O-octanoylglyceryl, 3-Ophospho-linked (PIP2), CAS 204858-53-7, purchased from Echelon Biosciences (Salt Lake City, UT). Recombinant human PI3Kδ protein (p110δ/p85α) was purchased from Eurofins (St. Charles, MO). ATP, MgCl ₂ , DTT, EDTA, MOPS, and CHAPS were purchased from SigmaAldrich (St. Louis, MO).

Kinase reactions were performed in Greiner Bio-one 384-well white polystyrene plates from Thermo Fisher Scientific in a final volume of 25 μl. The inhibitors were first serially diluted in DMSO and then added to the wells of the plate before adding other reaction components. 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 MgCl ₂ , 5 mM DTT, and CHAPS 0.03%. Reactions were initiated by the addition of ATP, and the final reaction mixture consisted of 20 µM PIP2, 2 µM ATP, 0.5 µCi [y- ^33P ]ATP, 3.4 nM PI3Kδ. The reaction mixtures were incubated for 120 min and the reaction was stopped by adding 40 μl of SPA beads suspended in quenching buffer: 163 mM potassium phosphate, pH 7.8, 20% glycerol, 25 mM EDTA. The final concentration of SPA beads was 1.0 mg/mL. After sealing the plates, the plates were shaken overnight at room temperature and centrifuged at 1500 rpm for 10 min. The radioactivity of the product was determined by scintillation counting on a Topcount instrument (PerkinElmer). Determination of IC ₅₀ was performed by fitting a curve of the percentage of control solvent activity versus the logarithm of inhibitor concentration using GraphPad Prism 6.0 software.

The compounds of Examples 1, 5, 6 and 9 were tested in the assays described in Examples A, B and C and found the IC ₅₀ values shown in Table 1. A below.

-

Claims (92)

Table A IC ₅₀ values __________________________________ Etc. No. ΡΙ3Κγ 1С ₅₀ (nM) ΡΙ3Κ6 1С ₅₀ (nM) PI3Ky_THP1_RPS6_ELISA ic ₅₀ (nM) 1 + + No. 5 + ++ #### 6 + ++ ## 9 + + ## + Refers to IC ₅₀ < 100 nM; ++ refers to IC ₅₀ < 500 nM; +++ refers to IC ₅₀ <2000 nM; ++++ refers to IC ₅₀ > 2000 nM. # refers to IC ₅₀ < 100 nM; # # refers to IC ₅₀ < 500 nM; # ## refers to IC ₅₀ <1000 nM; # ### refers to IC ₅₀ > 1000 nM. Various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from the preceding description. Such modifications are also included within the scope of the appended claims. Each reference, including all patents, patent applications and publications cited herein, is incorporated herein by reference in its entirety. CLAIM 1. Crystal form IA of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide , having a powder x-ray diffraction pattern including 4 or more of the following peaks, expressed in degrees 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°. 2. Crystal form IIA of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide , having a powder x-ray diffraction pattern including 4 or more of the following peaks, expressed in degrees 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°. 3. Crystal form IIIA of compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide , having a powder x-ray diffraction pattern including 4 or more of the following peaks, expressed in degrees 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°. 4. A crystalline form according to any one of claims 1 to 3, which is anhydrous and non-solvated. 5. The crystalline form of claim 1, having a powder X-ray diffraction pattern as shown in FIG. 1. 6. The crystalline form according to claim 1 or 5, having a DSC thermogram containing an endothermic peak having a maximum at 193°C. 7. The crystalline form according to any one of claims 1, 5 or 6, having such a differential scanning calorimetry (DSC) thermogram as shown in FIG. 2. 8. The crystalline form according to any one of claims 1 or 5 to 7, having a thermogravimetric analysis (TGA) result as shown in FIG. 3. 9. The crystalline form according to claim 2, having a powder X-ray diffraction pattern as shown in FIG. 4. 10. The crystalline form according to claim 2 or 9, having a DSC thermogram containing an endothermic peak having a maximum at 180°C. 11. The crystalline form according to any one of claims 2, 9 or 10, having such a differential scanning calorimetry (DSC) thermogram as shown in FIG. 5. 12. The crystalline form according to any one of claims 2 or 9 to 11, having a thermogravimetric analysis (TGA) result as shown in FIG. 6. 13. The crystalline form according to claim 3, having a powder X-ray diffraction pattern as shown in FIG. 7. 14. The crystalline form according to claim 3 or 13, having a DSC thermogram containing an endothermic peak having a maximum at 143°C. 15. The crystalline form according to any one of claims 3, 13 or 14, having such a differential scanning calorimetry (DSC) thermogram as shown in FIG. 8. 16. Crystalline form according to any one of claims 3 or 13-15, having such a thermal result - 50 044976 gravimetric analysis (TGA), which is shown in Fig. 9. 17. Crystal form IB of compound 8-αamino-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 a powder X-ray diffraction pattern including 4 or more of the following peaks expressed in degrees 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°. 18. Crystalline form IIB of compound 8-amino-K-(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 a powder x-ray diffraction pattern including 4 or more of the following peaks expressed in degrees 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°. 19. The crystalline form according to claim 17 or 18, which is anhydrous and unsolvated. 20. The crystalline form of claim 17, having a powder X-ray diffraction pattern as shown in FIG. 10. 21. The crystalline form according to claim 17 or 20, having a DSC thermogram containing an endothermic peak having a maximum at 174°C. 22. The crystalline form according to any one of claims 20 or 21, having a differential scanning calorimetry (DSC) thermogram as shown in FIG. eleven. 23. The crystalline form of claim 18, having a powder X-ray diffraction pattern as shown in FIG. 12. 24. The crystalline form according to claim 18 or 23, having a DSC thermogram containing an endothermic peak having a maximum at 165°C. 25. The crystalline form according to any one of claims 18, 23 or 24, having a differential scanning calorimetry (DSC) thermogram as shown in FIG. 13. 26. Crystalline form of the compound 8-amino-N-(2-hydroxy-2-methylpropyl)-3-(2-(methyl-d ₃ )-5(1,1,1-trifluoro-2-hydroxypropan-2-yl )phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, wherein the crystalline form is the IC form having a powder X-ray diffraction pattern including 4 or more of the following peaks expressed in degrees 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°. 27. The crystalline form according to claim 26, which is anhydrous and unsolvated. 28. The crystalline form of claim 26 or 27, having a powder X-ray diffraction pattern as shown in FIG. 14. 29. The crystalline form according to any one of claims 26 to 28, having a DSC thermogram containing an endothermic peak having a maximum at 179°C. 30. The crystalline form according to any one of claims 26 to 29, having a differential scanning calorimetry (DSC) thermogram as shown in FIG. 15. 31. Hydrobromic acid salt of 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxypropanamide. 32. The salt according to claim 31, which is a 1:1 stoichiometric ratio of 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)- 3,3,3-trifluoro-2-hydroxypropanamide and hydrobromic acid. 33. Salt according to claim 31 or 32, which is a crystalline form. 34. The crystalline form according to claim 33, which is a solvated crystalline form. 35. The crystalline form according to claim 34, which is a crystalline form of methanol solvate. 36. Pharmaceutical composition containing a crystalline form according to any one of claims 1-30 and 34, 35 or a salt according to any one of claims 31-33. 37. The composition according to claim 36, characterized in that said composition additionally contains at least one pharmaceutically acceptable carrier. 38. A method of inhibiting the activity of PI3Ky kinase, comprising contacting the kinase with a crystalline form according to any one of claims 1-30 and 34, 35 or a salt according to any one of claims 31-33. 39. The method of claim 38, wherein said crystalline form is a selective inhibitor of PI3Ky over one or more of PI3Ka, PI3Ke and PI3Kδ. 40. A method of treating a disease or disorder in a patient, wherein said disease or disorder is associated with abnormal expression of PI3Ky kinase, comprising administering to said patient a therapeutically effective amount of a crystalline form of any one of claims 1 to 30 and 34, 35 or a salt of any one of paragraphs 31-33. 41. The method of claim 40, wherein the disease or disorder is an autoimmune disease or disorder, a malignancy, a cardiovascular disease, or a neurodegenerative disease. 42. The method of claim 40, wherein the disease or disorder is cancer - 51 044976 lung (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, cancer stomach, seminoma, teratocarcinoma, astrocytoma, neuroblastoma, glioma or sarcoma. 43. The method according to claim 42, characterized in that the sarcoma is an Askin tumor, pampiniform sarcoma, chondrosarcoma, Ewing sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft tissue sarcoma, angiosarcoma, phyllodes cystosarcoma, dermatofibrosarcoma protuberans, desmoid tumor , desmoplastic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, hemangiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma or undifferentiated pleomorphic sarcoma. 44. The method of claim 40, 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-grained lymphocytic leukemia, mature peripheral T-cell neoplasm (PTCL), anaplastic large cell lymphoma (ALCL), or lymphoblastic lymphoma. 45. The method according to claim 44, characterized in that the neoplasm from mature peripheral T cells (PTCL) is T-cell prolymphocytic leukemia, T-cell granular lymphocytic leukemia, aggressive NK cell leukemia, mycosis fungoides/Sézary syndrome, anaplastic large cell T-cell lymphoma, enteropathic T-cell lymphoma, adult T-cell leukemia/lymphoma, or angioimmunoblastic T-cell lymphoma. 46. The method according to claim 44, characterized in that the anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL. 47. The method of claim 40, wherein the disease or disorder is Burkitt's lymphoma, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanthoma, lymphoplasmacytic lymphoma, extranodal marginal zone cell lymphoma, Waldenström's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosal associated lymphoid tissue (MALT) lymphoma, large B-cell lymphoma of the mediastinum or thymus, lymphogranulomatosis, marginal lymphoma nal zone of the spleen, primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma, also known as asymptomatic myeloma, monoclonal gammopathy of unknown etiology (MGUE) or diffuse large B-cell lymphoma. 48. The method of claim 47, wherein the non-Hodgkin's lymphoma (NHL) is relapsed NHL, refractory NHL, recurrent follicular NHL, indolent NHL (sNHL) or aggressive NHL (aNHL). 49. The method of claim 47, wherein the diffuse large B cell lymphoma is diffuse large B cell lymphoma of activated B cell type (ABC) or diffuse large B cell lymphoma of germinal center B cells (GCB). 50. The method according to claim 47, wherein the Burkitt lymphoma is endemic Burkitt lymphoma, sporadic Burkitt lymphoma or Burkitt lymphoma-like lymphoma. 51. The method of claim 40, 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 hypertrophy, myasthenia gravis, Sjogren's syndrome, osteoarthritis, restenosis or atherosclerosis. 52. The method according to claim 40, characterized in that the disease or disorder is cardiac hypertrophy, cardiomyocyte dysfunction, acute coronary syndrome, chronic obstructive pulmonary disease (COPD), chronic bronchitis, high blood pressure, ischemia, ischemia-reperfusion, vasoconstriction , anemia, bacterial infection, viral infection, graft rejection, kidney disease, anaphylactic shock, fibrosis, skeletal muscle atrophy, skeletal muscle hypertrophy, angiogenesis, sepsis, graft-versus-host disease, allogeneic or xenogeneic transplantation, glomerulosclerosis, progressive renal fibrosis, idiopathic thrombocytopenia purpura (ITP), autoimmune hemolytic anemia, vasculitis, systemic lupus erythematosus, lupus nephritis, pemphigus or membranous nephropathy. 53. The method according to claim 52, characterized in that idiopathic thrombocytopenic purpura (ITP) is recurrent ITP or refractory ITP. 54. The method according to claim 52, characterized in that the vasculitis is Behcet's disease, syndrome - 52 044976 Kogan's, giant cell arteritis, polymyalgia rheumatica (RPM), Takayasu's arteritis, Buerger's disease or thromboangiitis obliterans, central nervous system vasculitis, Kawasaki disease, polyarteritis nodosa, Churg-Strauss syndrome, mixed cryoglobulinemic vasculitis such as essential or caused by hepatitis C virus (HCV) ), Henoch-Schönlein purpura (HSP), hypersensitivity vasculitis, microscopic polyangiitis, Wegener's granulomatosis, or antineutrophil cytoplasmic antibody (ANCA)-associated systemic vasculitis (AASV). 55. The method of claim 40, wherein the disease or disorder is Alzheimer's disease, central nervous system injury, or stroke. 56. Method for obtaining the crystalline form of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2- hydroxypropanamide, which involves dissolving a compound in a solvent system to form a solution and crystallizing the compound from solution. 57. The method according to claim 56, characterized in that the solvent system is isopropyl acetate and heptane. 58. The method according to claim 56 or 57, characterized in that said process further includes heating the solution to a temperature of from 70 to 90°C. 59. The method according to claim 58, characterized in that said process further includes cooling the solution to room temperature. 60. The method according to claim 56, characterized in that the solvent system is methanol. 61. The method according to claim 56, characterized in that said process further includes heating the solution to a temperature of from 50 to 70°C. 62. The method according to claim 61, characterized in that said process further includes cooling the solution to room temperature. 63. Method for obtaining the crystalline form of the compound 8-amino-K-(2-hydroxy-2-methylpropyl)3-(2-methyl-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl) imidazo[1,2-a]pyrazine-6-carboxamide, comprising dissolving the compound in a solvent system to form a solution and crystallizing the compound from the solution. 64. The method according to claim 63, characterized in that said process further includes heating the solution to a temperature of 70 to 90°C. 65. The method according to claim 63, characterized in that said process further includes cooling the solution to room temperature. 66. The method according to any one of claims 63-65, characterized in that the solvent system is isopropyl acetate and heptane. 67. Method for obtaining the crystalline form of the compound 8-amino-K-(2-hydroxy-2-methylpropyl)3-(2-(methyl-d ₃ )-5-(1,1,1-trifluoro-2-hydroxypropane-2 -yl)phenyl)imidazo[1,2-a]pyrazine-6-carboxamide, comprising dissolving the compound in a solvent system to form a solution and crystallizing the compound from the solution. 68. The method according to claim 67, characterized in that said process further includes heating the solution to a temperature of 70 to 90°C. 69. The method according to claim 67, characterized in that said process further includes cooling the solution to room temperature. 70. The method according to any one of claims 67-69, characterized in that the solvent system is isopropyl acetate and heptane. 71. Method for preparing the hydrobromic acid salt of the compound 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro -2-hydroxypropanamide, comprising dissolving the compound in a solvent system to form a solution and adding hydrobromic acid to the solution. 72. The method according to claim 71, characterized in that the solvent system is methanol. 73. The method according to claim 71 or 72, characterized in that hydrobromic acid is added to the solution in the form of an aqueous solution of hydrobromic acid. 74. The method according to any one of claims 71-73, characterized in that an excess amount of hydrobromic acid is added to the solution per 1 equivalent. 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]πyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxyπpropanamide. 75. The method according to any one of claims 71-74, characterized in that from 1.1 to 1.5 equivalents are added to the solution. hydrobromic acid per 1 eq. 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2a]πyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2-hydroxyπpropanamide. 76. The method according to any one of claims 71-75, further comprising isolating the hydrobromic acid salt 2-(3-(8-amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4 -methylphenyl)-3,3,3trifluoro-2-hydroxypropanamide. 77. The method according to claim 76, characterized in that the hydrobromic acid salt is 2-(3-(8-amino-6(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3 ,3,3-trifluoro-2-hydroxypropanamide is isolated in crystalline form. 78. The method according to claim 76 or 77, characterized in that the hydrobromic acid salt is 2-(3-(8- - 53 044976 amino-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-3-yl)-4-methylphenyl)-3,3,3-trifluoro-2hydroxypropanamide is isolated in the form of a crystalline methanol solvate. 79. Use of a crystalline form according to any one of claims 1-30 and 34, 35 or a salt according to any one of claims 31-33 to inhibit the activity of PI3Ky kinase by contacting the kinase with a crystalline form according to any one of claims 1-30 and 34, 35 or salt according to any one of paragraphs 31-33. 80. Use according to claim 79, wherein said crystalline form is a selective inhibitor of PI3Ky over one or more of PI3Ka, PI3Ke and PI3Kδ. 81. Use of a crystalline form according to any one of claims 1-30 and 34, 35 or a salt according to any one of claims 31-33 for the treatment of a disease or disorder in a patient, wherein said disease or disorder is associated with abnormal expression or activity of PI3Ky kinase . 82. Use according to claim 81, wherein the disease or disorder is an autoimmune disease or disorder, a malignancy, a cardiovascular disease or a neurodegenerative disease. 83. The use of claim 81, wherein the disease or disorder is lung cancer (eg, non-small cell lung cancer), melanoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, colon cancer, endometrial cancer, urinary tract cancer bladder, skin cancer, uterine cancer, kidney cancer, stomach cancer, seminoma, teratocarcinoma, astrocytoma, neuroblastoma, glioma or sarcoma. 84. Use according to claim 83, where the sarcoma is Askin's tumor, pampiniform sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft tissue sarcoma, angiosarcoma, cystosarcoma phyllodes, dermatofibrosarcoma protuberans, desmoid tumor, desmoplast ic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, hemangiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoma or undifferentiated pleomorphic sarcoma. 85. The use of claim 81, 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-cell ALL), cutaneous T-cell lymphoma, large-grained lymphocytic leukemia, mature peripheral T-cell neoplasm (PTCL), anaplastic large cell lymphoma (ALCL), or lymphoblastic lymphoma. 86. Use according to claim 85, 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-type T-cell lymphoma, adult T-cell leukemia/lymphoma, or angioimmunoblastic T-cell lymphoma. 87. Use according to claim 85, wherein the anaplastic large cell lymphoma (ALCL) is systemic ALCL or primary cutaneous ALCL. 88. The use of claim 81, wherein the disease or disorder is Burkitt's lymphoma, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, mantle cell lymphoma, small lymphocytic lymphoma, follicular lymphoma, xeroderma pigmentosum, keratoacanthoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma, Waldenström's macroglobulinemia, prolymphocytic leukemia, acute lymphoblastic leukemia, myelofibrosis, mucosal associated lymphoid tissue (MALT) lymphoma, large B-cell lymphoma of the mediastinum or thymus, lymphogranulomatosis, splenic marginal zone lymphoma, primary effusion lymphoma, intravascular large B-cell lymphoma, plasma cell leukemia, extramedullary plasmacytoma, smoldering myeloma, also known as asymptomatic myeloma, monoclonal gammopathy of unknown etiology (MGUE), or diffuse large B-cell lymphoma. 89. The use of claim 88, wherein the non-Hodgkin's lymphoma (NHL) is relapsed NHL, refractory NHL, relapsed follicular NHL, indolent NHL (sNHL) or aggressive NHL (aNHL). 90. The use of claim 88, wherein the diffuse large B cell lymphoma is diffuse large B cell lymphoma of activated B cell type (ABC) or diffuse large B cell lymphoma of germinal center B cells (GCB). 91. Use according to claim 88, wherein the Burkitt's lymphoma is endemic Burkitt's lymphoma, sporadic Burkitt's lymphoma or Burkitt's lymphoma-like lymphoma. 92. Use according to claim 81, where the disease or disorder is rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, asthma, allergies, allergic rhinitis, pancreatitis, psoriasis, anaphylaxis, glomerulonephritis, inflammatory bowel disease, thrombosis, meninitis -