WO2022256550A1 - Crystalline forms of an adenosine a2b receptor antagonist - Google Patents

Abstract

Forms of ethyl 3-ethyl-2,6-dioxo-1-propyl-8-(1-(3-(trifluoromethyl)benzyl)-1H-pyrazol-4-yl)-1,2,3,6-tetrahydro-7H-purine-7-carboxylate, designated herein as Compound I, were prepared and characterized in the solid state. Also provided are processes of manufacture and methods of using the forms of Compound I.

Description

CRYSTALLINE FORMS OF AN ADENOSINE A2B RECEPTOR ANTAGONIST

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claim the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 63/197,128, filed June 4, 2021, which is incorporated herein by reference in its entirety.

FIELD

[0002] The present disclosure relates generally to solid forms of compounds that modulate the adenosine A2_B receptor, pharmaceutical compositions thereof, therapeutic uses thereof, and processes for making the solid forms.

BACKGROUND

[0003] The present disclosure relates to small molecule antagonists of adenosine A2_B receptor and their use as therapeutic agents, for example, in treating diseases such cancer. A2_B adenosine receptors play a role in signaling pathways.

[0004] Some A2_B adenosine receptor antagonists are relatively insoluble in aqueous media and/or difficult to formulate using conventional pharmaceutical excipients, and thus can be difficult to formulate in a manner that provides reproducible plasma levels of the compound in mammals, in particular humans. A need exists for improving the bioavailability A2_B adenosine receptor antagonists.

SUMMARY

[0005] The present disclosure provides polymorphic forms of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l- (3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7-carboxylate (Compound I) and salts, co-crystals, solvates, and hydrates thereof. Also described herein are processes for making the forms of Compound I, pharmaceutical compositions comprising forms of Compound I, and methods for using such forms and pharmaceutical compositions in the treatment of diseases mediated by adenosine A2_B receptors.

BRIEF DESCRIPTION OF THE DRAWINGS

[0006] The drawings described herein are for illustration purposes only. The drawings are not intended to limit the scope of the present disclosure.

[0007] FIG. 1 shows an X-Ray Powder Diffraction (XRPD) pattern of Compound I Form A.

[0008] FIG. 2 shows a thermogravimetric analysis (TGA) (top line) and a differential scanning calorimeter (DSC) curve (bottom line) of Compound I Form A.

[0009] FIG. 3 shows a dynamic vapor sorption (DVS isotherm plot) of Compound I Form A.

[0010] FIG. 4 shows an X-Ray Powder Diffraction (XRPD) pattern of Compound I Form B. [0011] FIG. 5 shows a thermogravimetric analysis (TGA) (top line) and a differential scanning calorimeter (DSC) curve (bottom line) of Compound I Form B.

[0012] FIG. 6 shows an X-Ray Powder Diffraction (XRPD) pattern of Compound II.

[0013] FIG. 7 shows a shows a thermogravimetric analysis (TGA) (top line) and a differential scanning calorimeter (DSC) curve (bottom line) of Compound II.

[0014] FIG. 8 shows an X-Ray Powder Diffraction (XRPD) pattern of Compound I crystallized from ethanol.

[0015] FIG. 9A shows a thermogravimetric analysis (TGA) and FIG. 9B shows a differential scanning calorimeter (DSC) curve of Compound I crystallized from ethanol.

[0016] FIG. 10 shows an ORTEP plot of Compound I crystallized from ethanol.

DETAILED DESCRIPTION

[0017] The compound ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH- pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7-carboxylate, designated herein as Compound I, has the following formula:

Compound I.

[0018] Compound I is an antagonist of adenosine A2_B receptors and is described as Compound O in PCT International Application Publication No. WO 2019/173380. The synthesis and method of use thereof is described in PCT International Application Publication No. WO 2019/173380 which is herein incorporated by reference in its entirety.

1. Definitions

[0019] As used in the present specification, the following words and phrases are generally intended to have the meanings as set forth below, except to the extent that the context in which they are used indicates otherwise.

[0020] The term “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” Further, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. Thus, reference to “the compound” includes a plurality of such compounds, and reference to “the assay” includes reference to one or more assays and equivalents thereof known to those skilled in the art. [0021] Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. In certain embodiments, the term “about” includes the indicated amount ± 10%. In other embodiments, the term “about” includes the indicated amount ± 5%.

In certain other embodiments, the term “about” includes the indicated amount ± 2.5%. In certain other embodiments, the term “about” includes the indicated amount ± 1%. Also, to the term “about X” includes description of “X”. In certain embodiments, a temperature of “about X °C” includes a temperature of X ± 2 °C. In certain embodiments, a temperature of “about X °C” includes a temperature of X ± 5 °C.

[0022] Recitation of numeric ranges of values throughout the disclosure is intended to serve as a shorthand notation of referring individually to each separate value falling within the range inclusive of the values defining the range, and each separate value is incorporated in the specification as it were individually recited herein.

[0023] Forms of Compound I or salts, co-crystals, solvates, or hydrates thereof are provided herein. In one embodiment, reference to a form of Compound I or a salt, co-crystal, solvate, or hydrate thereof means that at least 50% to 99% ( e.g ., at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) of Compound I or a salt, co-crystal, solvate, or hydrate thereof present in a composition is in the designated form. For instance, in one embodiment, reference to Compound I Form A means that at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of Compound I present in a composition is in Form A. For instance, in another embodiment, reference to Compound I Form B means that at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of Compound I present in a composition is in Form B.

[0024] The term “solid form” refers to a type of solid-state material that includes amorphous as well as crystalline forms. The term “crystalline form” refers to polymorphs as well as solvates, hydrates, etc. The term “polymorph” refers to a particular crystal structure having particular physical properties such as X-ray diffraction, melting point, and the like.

[0025] The term “solvate” refers to a complex formed by combination of solvent molecules with molecules or ions of the solute. The solvent can be an organic compound, an inorganic compound, or a mixture of both. As used herein, the term “solvate” includes a “hydrate” (i.e. , a complex formed by combination of water molecules with molecules or ions of the solute), hemi-hydrate, channel hydrate, etc. Some examples of solvents include, but are not limited to, methanol, N,N-dimethyIformamide, tetrahydrofuran, dime thylsulf oxide, and water. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present disclosure.

[0026] The term “co-crystal” refers to a molecular complex of a compound disclosed herein and one or more non-ionized co-crystal formers connected via non-covalent interactions. In some embodiments, the co-crystals disclosed herein may include a non-ionized form of Compound I (e.g., Compound I free form) and one or more non-ionized co-crystal formers, where non-ionized Compound I and the co-crystal former (s) are connected through non-covalent interactions. In some embodiments, co-crystals disclosed herein may include an ionized form of Compound I (e.g., a salt of Compound I) and one or more non- ionized co-crystals formers, where ionized Compound I and the co-crystal former(s) are connected through non-covalent interactions. Co-crystals may additionally be present in anhydrous, solvated or hydrated forms. In certain instances, co-crystals may have improved properties as compared to the parent form (i.e., the free molecule, zwitterion, etc.) or a salt of the parent compound. Improved properties can be increased solubility, increased dissolution, increased bioavailability, increased dose response, decreased hygroscopicity, increased stability, a crystalline form of a normally amorphous compound, a crystalline form of a difficult to salt or unsaltable compound, decreased form diversity, more desired morphology, and the like. Methods for making and characterizing co-crystals are known to those of skill in the art.

[0027] The term “co-crystal former” or “co-former” refers to one or more pharmaceutically acceptable bases or pharmaceutically acceptable acids disclosed herein in association with Compound I, or any other compound disclosed herein.

[0028] The term “amorphous” refers to a state in which the material lacks long range order at the molecular level and, depending upon temperature, may exhibit the physical properties of a solid or a liquid. Typically such materials do not give distinctive X-ray diffraction patterns and, while exhibiting the properties of a solid, are more formally described as a liquid. Upon heating, a change from solid to liquid properties occurs which is characterized by a change of state, typically second order (glass transition).

[0029] Any formula or structure given herein, including Compound I, is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. It is understood that for any given atom, the isotopes may be present essentially in ratios according to their natural occurrence, or one or more particular atoms may be enhanced with respect to one or more isotopes using synthetic methods known to one skilled in the art. Thus, hydrogen includes for example 'H, ¾, ³H; carbon includes for example ⁿC, ¹²C, ¹³C, ¹⁴C; oxygen includes for example ¹⁶0, ¹⁷0, ¹⁸0; nitrogen includes for example ¹³N, ¹⁴N, ¹⁵N; sulfur includes for example ³²S, ³³S, ³⁴S, ³⁵S, ³⁶S, ³⁷S, ³⁸S; fluoro includes for example ¹⁷F, ¹⁸F, ¹⁹F; chloro includes for example ³⁵C1, ³⁶C1, ³⁷C1, ³⁸C1, ³⁹C1; and the like.

[0030] As used herein, the terms “treat,” “treating,” “therapy,” “therapies,” and like terms refer to the administration of material, e.g., any one or more solid, crystalline or polymorphs of Compound I as described herein in an amount effective to prevent, alleviate, or ameliorate one or more symptoms of a disease or condition, i.e., indication, and/or to prolong the survival of the subject being treated.

[0031] The term “administering” refers to oral administration, administration as a suppository, topical contact, intravenous, intraperitoneal, intramuscular, intralesional, intranasal or subcutaneous administration, or the implantation of a slow-release device e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc.

[0032] As used herein, the term “modulating” or “modulate” refers to an effect of altering a biological activity, especially a biological activity associated with a particular biomolecule such as adenosine A_2B receptor. For example, an agonist or antagonist of a particular biomolecule modulates the activity of that biomolecule, e.g., adenosine A_2B receptor, by either increasing (e.g. agonist, activator), or decreasing (e.g. antagonist, inhibitor) the activity of the biomolecule. Such activity is typically indicated in terms of an inhibitory concentration (IC₅₀) or excitation concentration (EC₅₀) of the compound for an inhibitor or activator, respectively, with respect to, for example, adenosine A_2B receptor.

[0033] As used herein, the term “adenosine A_2B receptor mediated disease or condition,” refers to a disease or condition in which the biological function of adenosine A_2B receptor, including any mutations thereof, affects the development, course, and/or symptoms of the disease or condition, and/or in which modulation of adenosine A_2B receptor alters the development, course, and/or symptoms of the disease or condition. The adenosine A_2B receptor mediated disease or condition includes a disease or condition for which adenosine A_2B receptor modulation provides a therapeutic benefit, e.g. wherein treatment with compound(s), including one or more solid, crystalline or polymorphs of Compound I as described herein, provides a therapeutic benefit to the subject suffering from or at risk of the disease or condition.

[0034] As used herein, the term “composition” refers to a pharmaceutical preparation suitable for administration to an intended subject for therapeutic purposes that contains at least one pharmaceutically active compound, including any solid form thereof. The composition may include at least one pharmaceutically acceptable component to provide an improved formulation of the compound, such as a suitable carrier or excipient.

[0035] As used herein, the term “subject” or “patient” refers to a living organism that is treated with compounds as described herein, including, but not limited to, any mammal, such as a human, other primates, sports animals, animals of commercial interest such as cattle, farm animals such as horses, or pets such as dogs and cats.

[0036] The term “pharmaceutically acceptable” indicates that the indicated material does not have properties that would cause a reasonably prudent medical practitioner to avoid administration of the material to a patient, taking into consideration the disease or conditions to be treated and the respective route of administration. For example, it is commonly required that such a material be essentially sterile, e.g., for injectables. [0037] In the present context, the term “therapeutically effective” or “effective amount” indicates that the materials or amount of material is effective to prevent, alleviate, or ameliorate one or more symptoms of a disease or medical condition, and/or to prolong the survival of the subject being treated. The therapeutically effective amount will vary depending on the compound, the disorder or condition and its severity and the age, weight, etc., of the mammal to be treated. For example, an effective amount is an amount sufficient to effectuate a beneficial or desired clinical result. The effective amounts can be provided all at once in a single administration or in fractional amounts that provide the effective amount in several administrations. The precise determination of what would be considered an effective amount may be based on factors individual to each subject, including their size, age, injury, and/or disease or injury being treated, and amount of time since the injury occurred or the disease began. One skilled in the art will be able to determine the effective amount for a given subject based on these considerations which are routine in the art.

[0038] In some embodiments, the phrase “substantially shown in Figure” as applied to an X-ray powder diffractogram is meant to include a variation of ± 0.2 °2Q or ± 0.1 °2Q, as applied to DSC thermograms is meant to include a variation of ± 3 °Celsius, and as applied to thermogravimetric analysis (TGA) is meant to include a variation of ± 2% in weight loss.

[0039] “Substantially pure form (of a polymorph),” in some embodiments, means that in the referenced material, at least 99.9% of the material is the referenced polymorph. “Substantially pure form (of a polymorph),” in some embodiments, means that in the referenced material, at least 99.5% of the material is the referenced polymorph. “Substantially pure form (of a polymorph),” in some embodiments, means that in the referenced material, at least 99% of the material is the referenced polymorph. “Substantially pure form (of a polymorph),” in some embodiments, means that in the referenced material, at least 98% of the material is the referenced polymorph. “Substantially pure form (of a polymorph),” in some embodiments, means that in the referenced material, at least 97% of the material is the referenced polymorph. “Substantially pure form (of a polymorph),” in some embodiments, means that in the referenced material, at least 96% of the material is the referenced polymorph. “Substantially pure form (of a polymorph),” in some embodiments, means that in the referenced material, at least 95% of the material is the referenced polymorph. In the context of the use, testing, or screening of compounds that are or may be modulators, the term “contacting” means that the compound(s) are caused to be in sufficient proximity to a particular molecule, complex, cell, tissue, organism, or other specified material that potential binding interactions and/or chemical reaction between the compound and other specified material can occur.

[0040] In addition, abbreviations as used herein have respective meanings as follows:

2. Forms of Compound I

[0041] As described generally above, the present disclosure provides crystalline forms of the compound, ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6- tetrahydro-7H-purine-7-carboxylate (hereinafter “compound” or “Compound I”), and salts, co-crystals, solvates, or hydrates thereof. Crystalline forms of Compound I and salts, co-crystals, solvates, or hydrates thereof, and other forms (e.g., amorphous forms) of Compound I and salts, co-crystals, solvates, or hydrates thereof are collectively referred to herein as “forms of Compound I.”

[0042] In some embodiments, Compound I is a free base. In some embodiments, Compound I is a salt or a co-crystal. In some embodiments, Compound I is a pharmaceutically acceptable salt or co crystal. In some embodiments, Compound I is a solvate. In some embodiments, Compound I is a hydrate. In some embodiments, Compound I is an anhydrate. [0043] In some embodiments, Compound I is an amorphous form.

CompoundIForm A

[0044] In one aspect, provided herein is a crystalline form of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l- (3-(trifluoromethyl)benzyl)-lF[-pyrazol-4-yl)-l,2,3,6-tetrahydro-7F[-purine-7-carboxylate (Compound I):

wherein an X-ray powder diffraction pattern of Compound I comprises the following peaks: 15.23°±0.2°, 21.57°±0.2° and 22.43 °±0.2° (Compound I, Form A), expressed as 20 angles determined using Cu K-a (l = 1.5418 A) radiation.

[0045] In some embodiments, the X-ray powder diffraction pattern of Compound I, Form A, further comprises one or more peaks selected from 16.66°±0.2°, 18.79°±0.2°, and 23.40°±0.2°, expressed as 20 angles determined using Cu K-a (l = 1.5418Ά) radiation.

[0046] In some embodiments, the X-ray powder diffraction pattern of Compound I, Form A, further comprises two or more peaks selected from 6.26°±0.2°, 16.66°±0.2°, 18.31°±0.2°, 18.79°±0.2°, 23.40°±0.2° and 25.18 °±0.2°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation. In some embodiments, the X-ray powder diffraction pattern of Compound I, Form A, comprises three or more peaks selected from 6.26°±0.2°, 15.23°±0.2°, 16.66°±0.2°, 18.31°±0.2°, 18.79°±0.2°, 21.57°±0.2°, 22.43 °±0.2°, 23.40°±0.2°, and 25.18°±0.2°. In some embodiments, the X-ray powder diffraction pattern of Compound I, Form A, comprises four or more peaks selected from 6.26°±0.2°, 15.23°±0.2°, 16.66°±0.2°, 18.31°±0.2°, 18.79°±0.2°, 21.57°±0.2°, 22.43°±0.2°, 23.40°±0.2°, and 25.18°±0.2°. In some embodiments, the X-ray powder diffraction pattern of Compound I, Form A, comprises peaks at 6.26°±0.2°, 12.12°±0.2°, 12.50°±0.2°, 15.23°±0.2°, 16.66°±0.2°, 18.31°±0.2°, 18.79°±0.2°, 21.57°±0.2°, 22.43 °±0.2°, 23.40°±0.2°, and 25.18°±0.2°.

[0047] In some embodiments, the diffraction pattern of Compound I, Form A, is substantially free of peaks at 18.15°±0.05°, and 22.76°±0.05°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation.

[0048] In some embodiments, the diffraction pattern of Compound I, Form A, substantially as shown in Figure 1. In some embodiments, Compound I, Form A, has a differential scanning calorimetry (DSC) curve that shows an endotherm onset at about 131.9 °C. In some embodiments, Compound I, Form A, has a DSC curve substantially as shown in Figure 2. In some embodiments, Compound I, Form A, has a dynamic vapor sorption (DVS) isotherm substantially as shown in Figure 3.

[0049] In some embodiments, Compound I, Form A, is an anhydrate. [0050] In some embodiments, Compound I, Form A, is prepared by addition of water to a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3- (trifluoromethyl)benzyl)-lFl-pyrazol-4-yl)-l,2,3,6-tetrahydro-7F[-purine-7-carboxylate in acetone or tetrahydrofuran; or by addition of n-heptane to a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3- (trifluoromethyl)benzyl)-lFl-pyrazol-4-yl)-l,2,3,6-tetrahydro-7F[-purine-7-carboxylate in dichloromethane or ethyl acetate.

[0051] In some embodiments, Compound I, Form A, is prepared by slow cooling or slow evaporation of a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH- pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7-carboxylate.

[0052] In some embodiments, Compound I, Form A, is prepared from a slurry of ethyl 3-ethyl-2,6- dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7- carboxylate in a solvent, wherein the slurry is maintained at room temperature.

[0053] In some embodiments, the solvent for slow cooling, slow evaporation, or slurry, is selected from ethanol, methyl-tert-butyl ether, dimethyl sulfoxide, dichloromethane, acetone, ethyl acetate, isopropyl acetate, toluene, isopropyl alcohol, methanol, tetrahydrofuran, n-heptane, water, or any combination thereof.

[0054] In some embodiments, Compound I Form A is crystallized by slow evaporation of a solution of Compound I in ethanol at about 4 °C. In some embodiments, Compound I Form A is prepared as described in Example 2 and Example 6.

Compound I Form B

[0055] In another aspect, provided herein is a crystalline form of ethyl 3-ethyl-2,6-dioxo-l-propyl-8- (l-(3-(trifluoromethyl)benzyl)-lFl-pyrazol-4-yl)-l,2,3,6-tetrahydro-7F[-purine-7-carboxylate (Compound

I):

wherein an X-ray powder diffraction pattern of Compound I comprises the following peaks: 12.34°±0.2°, 18.15°±0.2°, and 22.76°±0.2°, (Compound I, Form B), expressed as 2Q angles determined using Cu K-a (l = 1.5418 A) radiation. [0056] In some embodiments, the X-ray powder diffraction pattern of Compound I, Form B, further comprises one or more peaks selected from 15.56°±0.2°, and 21.04°±0.2°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation.

[0057] In some embodiments, the X-ray powder diffraction pattern of Compound I, Form B, further comprises two or more peaks selected from 6.18 °±0.2°, 15.56°±0.2°, 21.04°±0.2°, and 22.76°±0.2°, expressed as 2Q angles determined using Cu K-a (l = 1.5418A) radiation. In some embodiments, the X- ray powder diffraction pattern of Compound I, Form B, comprises three or more peaks selected from 6.18°±0.2°, 12.34°±0.2°, 15.56°±0.2°, 18.15°±0.2°, 21.04°±0.2°, and 22.76°±0.2°. In some embodiments, the X-ray powder diffraction pattern of Compound I, Form B, comprises four or more peaks selected from 6.18°±0.2°, 12.34°±0.2°, 15.56°±0.2°, 18.15°±0.2°, 21.04°±0.2°, and 22.76°±0.2°.

In some embodiments, the X-ray powder diffraction pattern of Compound I, Form B, comprises peaks at 6.18°±0.2°, 10.99°±0.2°, 11.33°±0.2°, 12.34°±0.2°, 15.56°±0.2°, 18.15°±0.2°, 21.04°±0.2°, and 22.76°±0.2°.

[0058] In some embodiments, the diffraction pattern of Compound I, Form B, is substantially free of peaks at 21.57°±0.05°, and 22.43 °±0.05°.

[0059] In some embodiments, the diffraction pattern of Compound I, Form B, is substantially as shown in Figure 4. In some embodiments, Compound I, Form B, is characterized by a differential scanning calorimetry (DSC) curve that shows endotherm onsets at about 119.4 °C and at 130.9 °C. In some embodiments, Compound I, Form B, has a DSC curve substantially as shown in Figure 5.

[0060] In some embodiments, Compound I, Form B, is an anhydrate.

[0061] In some embodiments, Compound I, Form B, is prepared by addition of water to a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6- tetrahydro-7H-purine-7-carboxylate in N-methyl pyrrolidine.

[0062] In some embodiments, Compound I, Form B, is prepared by evaporation of a slurry of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H- purine-7-carboxylate in a mixture of acetonitrile and isopropyl alcohol, wherein the slurry is maintained at a temperature ranging from - 20 °C to 5 °C. In some embodiments, Compound I Form B is prepared as described in Example 3.

[0063] In some embodiments, heating a slurry of Compound I in isopropanol, or a 1:9 mixture of isopropyl acetate/isopropanol to 50 °C provides a mixture of Compound I Form A and Compound II. In some embodiment, slow evaporation of a solution of Compound I in methanol at room temperature provides a mixture of Compound I Form A and Compound II.

[0064] In some embodiments, crystalline Compound II is prepared as described in Example 4.

3. Pharmaceutical Compositions, Kits, and Modes of Administration

[0065] The forms of Compound I and/or Compound II as described herein may be administered in a pharmaceutical composition. Thus, provided herein are pharmaceutical compositions comprising one or more of the forms of Compound I and/or Compound II described herein and one or more pharmaceutically acceptable vehicles such as carriers, adjuvants and excipients. Suitable pharmaceutically acceptable vehicles may include, for example, inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants. Such compositions are prepared in a manner well known in the pharmaceutical art. See, e.g., Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (G.S. Banker & C.T. Rhodes, Eds.). The pharmaceutical compositions may be administered alone or in combination with other therapeutic agents.

[0066] Some embodiments are directed to pharmaceutical compositions comprising a therapeutically effective amount of a solid form of Compound I described herein. In some embodiments, a pharmaceutical composition comprises a solid form selected from Compound I Form A and Compound

I Form B; and one or more pharmaceutically acceptable carriers. Some embodiments are directed to pharmaceutical compositions comprising a therapeutically effective amount of a solid form of Compound

II described herein. In some embodiments, a pharmaceutical composition comprises a solid form of Compound II described herein; and one or more pharmaceutically acceptable carriers.

[0067] Some embodiments are directed to pharmaceutical compositions comprising a crystalline form or amorphous form of Compound I as described herein and one or more pharmaceutically acceptable carriers. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 95% of Compound I is in a crystalline form as described herein. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 95% of Compound I is in an amorphous form as described herein. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 95% of Compound I is in Form A. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 95% of Compound I is in Form B. In one embodiment, a pharmaceutical composition comprises Compound II, wherein at least 95% of Compound II is in a form described in Example 4 and FIG. 6.

[0068] In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 97% of Compound I is in a crystalline form or an amorphous form as described herein. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 97% of Compound I is in Form A. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 97% of Compound I is in Form B. In one embodiment, a pharmaceutical composition comprises Compound II, wherein at least 97% of Compound II is in a form described in Example 4 and FIG. 6.

[0069] In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99% of Compound I is in a crystalline form as described herein. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99% of Compound I is in Form A. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99% of Compound I is in Form B. In one embodiment, a pharmaceutical composition comprises Compound II, wherein at least 99% of Compound II is in a form described in Example 4 and FIG. 6.

[0070] In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99.5% of Compound I is in a crystalline form as described herein. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99.5% of Compound I is in Form A. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99.5% of Compound I is in Form B. In one embodiment, a pharmaceutical composition comprises Compound II, wherein at least 99.5% of Compound II is in a form described in Example 4 and FIG. 6.

[0071] In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99.9% of Compound I is in a crystalline form as described herein. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99.9% of Compound I is in Form A. In one embodiment, a pharmaceutical composition comprises Compound I, wherein at least 99.9% of Compound I is in Form B. In one embodiment, a pharmaceutical composition comprises Compound II, wherein at least 99.9% of Compound II is in a form described in Example 4 and FIG. 6.

[0072] In some embodiments, compositions comprise pharmaceutically acceptable carriers or excipients, such as fillers, binders, disintegrants, glidants, lubricants, complexing agents, solubilizers, and surfactants, which may be chosen to facilitate administration of the compound by a particular route. Examples of carriers include calcium carbonate, calcium phosphate, various sugars such as lactose, glucose, or sucrose, types of starch, cellulose derivatives, gelatin, lipids, liposomes, nanoparticles, and the like. Carriers also include physiologically compatible liquids as solvents or for suspensions, including, for example, sterile solutions of water for injection (WFI), saline solution, dextrose solution, Hank’s solution, Ringer’s solution, vegetable oils, mineral oils, animal oils, polyethylene glycols, liquid paraffin, and the like. Excipients may also include, for example, colloidal silicon dioxide, silica gel, talc, magnesium silicate, calcium silicate, sodium aluminosilicate, magnesium trisilicate, powdered cellulose, macrocrystalline cellulose, carboxymethyl cellulose, cross-linked sodium carboxymethylcellulose, sodium benzoate, calcium carbonate, magnesium carbonate, stearic acid, aluminum stearate, calcium stearate, magnesium stearate, zinc stearate, sodium stearyl fumarate, syloid, stearowet C, magnesium oxide, starch, sodium starch glycolate, glyceryl monostearate, glyceryl dibehenate, glyceryl palmitostearate, hydrogenated vegetable oil, hydrogenated cotton seed oil, castor seed oil mineral oil, polyethylene glycol (e.g. PEG 4000-8000), polyoxyethylene glycol, poloxamers, povidone, crospovidone, croscarmellose sodium, alginic acid, casein, methacrylic acid divinylbenzene copolymer, sodium docusate, cyclodextrins (e.g. 2-hydroxypropyl-.delta.-cyclodextrin), polysorbates (e.g. polysorbate 80), cetrimide, TPGS (d-alpha-tocopherol polyethylene glycol 1000 succinate), magnesium lauryl sulfate, sodium lauryl sulfate, polyethylene glycol ethers, di-fatty acid ester of polyethylene glycols, or a polyoxyalkylene sorbitan fatty acid ester (e.g., polyoxyethylene sorbitan ester Tween®), polyoxyethylene sorbitan fatty acid esters, sorbitan fatty acid ester, e.g. a sorbitan fatty acid ester from a fatty acid such as oleic, stearic or palmitic acid, mannitol, xylitol, sorbitol, maltose, lactose, lactose monohydrate or lactose spray dried, sucrose, fructose, calcium phosphate, dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate, dextrates, dextran, dextrin, dextrose, cellulose acetate, maltodextrin, simethicone, polydextrosem, chitosan, gelatin, HPMC (hydroxypropyl methyl celluloses), HPC (hydroxypropyl cellulose), hydroxyethyl cellulose, and the like.

[0073] Pharmaceutical formulations may be presented in unit dose forms containing a predetermined amount of active ingredient per unit dose. Such a unit may contain, for example, 0.5 mg to 1 g, preferably 1 mg to 700 mg, more preferably 5 mg to 100 mg of a compound of the present disclosure (as a free-acid, solvate (including hydrate) or salt, in any form), depending on the condition being treated, the route of administration, and the age, weight and condition of the patient. Preferred unit dosage formulations are those containing a daily dose, weekly dose, monthly dose, a sub-dose or an appropriate fraction thereof, of an active ingredient. Furthermore, such pharmaceutical formulations may be prepared by any of the methods well known in the pharmacy art.

[0074] Compound I, and any of its forms as described herein, are usually administered in the form of pharmaceutical compositions. Thus, provided herein are also pharmaceutical compositions that contain one or more of Compound I, and any of its forms as described herein, a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers or prodrug thereof and one or more pharmaceutically acceptable vehicles selected from carriers, adjuvants and excipients. Suitable pharmaceutically acceptable vehicles may include, for example, inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants. Such compositions are prepared in a manner well known in the pharmaceutical art. See, e.g., Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (G.S. Banker & C.T. Rhodes, Eds.).

[0075] The pharmaceutical compositions may be administered in either single or multiple doses.

The pharmaceutical composition may be administered by various methods including, for example, rectal, buccal, intranasal and transdermal routes. In certain embodiments, the pharmaceutical composition may be administered by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant.

[0076] One mode for administration is parenteral, for example, by injection. The forms in which the pharmaceutical compositions described herein may be incorporated for administration by injection include, for example, aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles.

[0077] Oral administration may be another route for administration of the compounds described herein. Administration may be via, for example, capsule or enteric coated tablets. In making the pharmaceutical compositions that include at least one compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers or prodrug thereof, the active ingredient is usually diluted by an excipient and/or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be in the form of a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders.

[0078] Some examples of suitable excipients include, e.g., lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup and methyl cellulose. The formulations can additionally include lubricating agents such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl and propylhydroxy- benzoates; sweetening agents; and flavoring agents.

[0079] The compositions that include at least one of the forms of Compound I as described herein, can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the subject by employing procedures known in the art. Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Another formulation for use in the methods disclosed herein employ transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds described herein in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.

[0080] For preparing solid compositions such as tablets, the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of Compound I, and any of its forms as described herein. When referring to these preformulation compositions as homogeneous, the active ingredient may be dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. [0081] The tablets or pills of Compound I, and any of its for s as described herein, may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach. For example, the tablet or pill can include an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol and cellulose acetate.

[0082] In another aspect, the present disclosure provides kits or containers that include a Compound I, and any of its forms as described herein, or any of the pharmaceutical compositions thereof described herein. In some embodiments, the compound or composition is packaged, e.g., in a vial, bottle, flask, which may be further packaged, e.g., within a box, envelope, or bag; the compound or composition is approved by the U.S. Food and Drug Administration or similar regulatory agency for administration to a mammal, e.g., a human; the compound or composition is approved for administration to a mammal, e.g., a human, for a bromodomain protein mediated disease or condition; the kit or container disclosed herein may include written instructions for use and/or other indication that the compound or composition is suitable or approved for administration to a mammal, e.g., a human, for a bromodomain-mediated disease or condition; and the compound or composition may be packaged in unit dose or single dose form, e.g., single dose pills, capsules, or the like.

[0083] The amounts of various compounds to be administered can be determined by standard procedures taking into account factors such as the compound activity (in vitro, e.g. the compound IC50 vs. target, or in vivo activity in animal efficacy models), pharmacokinetic results in animal models (e.g. biological half-life or bioavailability), the age, size, and weight of the subject, and the disorder associated with the subject. The importance of these and other factors are well known to those of ordinary skill in the art. Generally, a dose will be in the range of about 0.01 to 50 mg/kg, also about 0.1 to 20 mg/kg of the subject being treated. Multiple doses may be used.

4. Dosing

[0084] The specific dose level of Compound I, and any of its forms as described herein, for any particular subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease in the subject undergoing therapy. For example, a dosage may be expressed as a number of milligrams of a compound described herein per kilogram of the subject’s body weight (mg/kg). Dosages of between about 0.1 and 150 mg/kg may be appropriate. In some embodiments, about 0.1 and 100 mg/kg may be appropriate. In other embodiments a dosage of between 0.5 and 60 mg/kg may be appropriate. In some embodiments, a dosage of from about 0.0001 to about 100 mg per kg of body weight per day, from about 0.001 to about 50 mg of compound per kg of body weight, or from about 0.01 to about 10 mg of compound per kg of body weight may be appropriate. Normalizing according to the subject’s body weight is particularly useful when adjusting dosages between subjects of widely disparate size, such as occurs when using the drug in both children and adult humans or when converting an effective dosage in a non-human subject such as dog to a dosage suitable for a human subject.

5. Disease indications and modulation of Adenosine A ₂₁s receptor

[0085] Compound I is a prodrug of Compound II. In some cases, prodrugs such as Compound I can be hydrolyzed by esterases (e.g., in gastrointestinal tract and/or in blood) and converted into Compound II in an aqueous solution. In some cases, acid labile prodrugs such as Compound I can be converted into Compound II in an acidic environment (e.g., in the stomach). In some cases, prodrugs, which are stable in the acidic environment and/or stable against hydrolysis by esterases, may not be a good prodrug candidate for Compound II. In some cases, formulation and/or bioavailability of the prodrug, i.e., Compound I may depend on the solid form of Compound I used.

[0086] In one aspect, the compounds, compositions, and/or formulations disclosed herein can be used to treat cancer. On endothelial cells, for example, adenosine can bind to the A_2B adenosine receptors, thereby stimulating angiogenesis. On T cells, A_2B adenosine receptor stimulation can lead to type I protein kinase A (PKA) isoform activation that can hamper T cell activation through inhibition of T-cell antigen receptor (TCR) proximal kinases Lck and Fyn. The pro-metastatic Fra-1 transcription factor can also induce A_2B adenosine receptor expression on cancer cells, and thus A_2B adenosine receptor antagonist can inhibit metastasis of Fra-1 -expressing cells. A_2B adenosine receptor signaling activation can impair antigen presentation and can also inhibit signal transducer and activator of transcription 1 (STAT1) activation. The diversity of signaling and biological activities of A_2B adenosine receptor can render it an attractive cancer target to promote anti-tumor immunity and suppress tumor cell metastasis.

[0087] In another aspect, the compounds, compositions, and/or formulations disclosed herein can be used to treat fibrosis. A commonly ingested adenosine receptor antagonist, caffeine, can block the development of hepatic fibrosis, an effect that may explain the epidemiologic finding that coffee drinking, in a dose-dependent fashion, can reduce the likelihood of death from liver disease. A_2B adenosine receptors can also play a role in the pathogenesis of interstitial fibrosis. Adenosine, acting at A_2B adenosine receptors, can stimulate hepatic stellate cell-mediated fibrosis of the liver by increasing production of collagen I and III via two distinct mitogen-activated protein kinase (MAPK) -dependent pathways, extracellular signal-regulated kinase 1/2 (ERK1/2) and p38MAPK, respectively. Over activation of A_2B adenosine receptors can be involved in liver, lung and heart fibrosis. Accordingly, A_2B adenosine receptors may be a good therapeutic target for fibrosis of the liver, lungs, heart, and/or skin.

[0088] In another aspect, the compounds, compositions, and/or formulations disclosed herein can be used to treat diabetes and/or obesity. Insensitivity to insulin can exacerbate diabetes and/or obesity. Insulin sensitivity can be decreased by the interaction of adenosine with A_2B adenosine receptors. Thus, blocking the A2_B adenosine receptors of individuals with diabetes and/or obesity can benefit patients with these disorders.

[0089] In another aspect, the compounds, compositions, and/or formulations disclosed herein can be used to treat neurological disorders, such as dementias and Alzheimer's disease. Adenosine acting at A2_B adenosine receptors can over-stimulate cerebral interleukin 6 (IL-6), a cytokine associated with dementias and Alzheimer's disease. Inhibiting the binding of adenosine to A2_B adenosine receptors can therefore mitigate those neurological disorders that are produced by IL-6.

[0090] In another aspect, the compounds, compositions, and/or formulations disclosed herein can be used to treat type I hypersensitivity disorders, such as chronic obstructive pulmonary disease (COPD), asthma, hay fever, and atopic eczema. These type I hypersensitivity disorders can be stimulated by mast cells binding to A2_B adenosine receptors. Therefore, blocking A2_B adenosine receptors can provide a therapeutic benefit against such disorders.

[0091] In some embodiments, provided herein is a method for treating a disease or condition mediated, at least in part, by the A2_B adenosine receptor in a mammal, the method comprising administering to the mammal a therapeutically effective amount of any crystalline form of Compound I described herein, or any composition described herein.

[0092] In some embodiments, the disease or condition is selected from the group consisting of cardiovascular diseases, fibrosis, neurological disorders, type I hypersensitivity disorders, chronic and acute liver diseases, lung diseases, renal diseases, diabetes, obesity, and cancer. In some embodiments, the cancer is a hormone-related cancer. In some embodiments, the hormone -related cancer is breast cancer, endometrial cancer, ovarian cancer, prostate cancer, testicular cancer, thyroid cancer or osteosarcoma. In some embodiments, the hormone-related cancer is metastatic castration resistant prostate cancer. In some embodiments, the hormone -related cancer is breast cancer.

[0093] In certain embodiments, the present disclosure provides use of Compound I, and any of its forms as described herein, or any of the pharmaceutical compositions thereof described herein in the manufacture of a medicament for the treatment of a disease or condition as described herein. In other embodiments, the present disclosure provides Compound I, and any of its forms as described herein, or any of the pharmaceutical compositions thereof described herein for use in treating a disease or condition as described herein.

[0094] Provided is the use of any crystalline form of Compound I described herein, or any composition described herein, for treating a disease or condition mediated, at least in part, by the A2_B adenosine receptor in a mammal. Provided is the use of any crystalline form of Compound I described herein, or any composition described herein, for the manufacture of a medicament for treating a disease or condition mediated, at least in part, by the A2_B adenosine receptor in a mammal. EXAMPLES

Example 1. Polymorph screening of Compound I

[0095] Compound I was prepared as described in Example 16 of WO 2019/173380. Polymorph screening experiments were performed via anti-solvent addition, slurry, slow evaporation and slow cooling.

[0096] Table 1-1 below provides a summary of anti-solvent addition experiments. Approximately 15 mg of starting material (Compound I) was dissolved in 0.15-1.5 mL corresponding solvent to obtain a clear solution. The solution was magnetically stirred with addition of anti-solvent until precipitates appeared or the total volume of anti-solvent reached 10 mL. The obtained precipitates were isolated for XRPD analysis.

Table 1-1

*Clear solution was obtained after anti-solvent addition and at 5 °C and -20 °C, which was allowed to evaporate at RT.

[0097] Table 1-2 below provides a summary of slurry conversion experiments at room temperature. Approximately 15 mg of starting material (Compound I) was suspended in 0.5 mL of corresponding solvent in an HPLC vial. After the suspension was stirred magnetically for 3 days at RT, the remaining solids were isolated for XRPD analysis.

Table 1-2

[0098] Table 1-3 below provides a summary of slurry conversion experiments at 50 °C. Approximately 15 mg of starting material (Compound I) was suspended in 0.3 mL of corresponding solvent in an HPLC vial. After the suspension was magnetically stirred for about 4 days at 50 °C, the remaining solids were isolated for XRPD analysis.

Table 1-3

[0099] Table 1-4 below provides a summary of slow evaporation experiments. Approximately 15 mg of starting material (Compound I) was dissolved in 0.2-2.0 mL corresponding solvent in a 3-mL glass vial. If not dissolved completely, suspensions were filtered using a 0.45 pm PTFE membrane and the filtrates were used for the follow-up steps. The visually clear solutions were subjected to evaporation at RT in vials sealed by Parafilm® (poke 5 small holes). The solids were isolated for XRPD analysis.

Table 1-4

[0100] Table 1-5 below provides a summary of slow cooling experiments. Approximately 15 mg of starting material (Compound I) was suspended in 0.5-1.5 mL of corresponding solvent in a 3-mL glass vial at RT. The suspension was then heated to 50 °C, equilibrated for about two hours and filtered to a new vial using a 0.45 pm PTLE membrane. Liltrates were slowly cooled down to 5 °C at a rate of 0.1 °C/min. The obtained solids were kept isothermal at 5 °C before isolation for XRPD analysis.

Table 1-5

* Clear solution was obtained after slow cooling and further storage at - 20 °C, which was transferred to evaporate at RT.

[0101] Two forms of Compound I, Form A and Form B, were obtained during screening. Example 2. Compound I Form A

[0102] FIG. 1 shows the XRPD spectrum for Form A. For XRPD analysis, a PANalytical X’Pert3 X-ray powder diffract meter was used. The XRPD parameters used are listed in Table 2-1.

Table 2-1

[0103] TGA data were collected using a TA Q5000/ Discovery 5500 TGA from TA Instruments. DSC was performed using a TA Discovery 2500 DSC from TA Instruments. Table 2-2 provides the parameters for TGA/DSC.

Table 2-2

[0104] TGA and DSC results for Compound I Form A are displayed in FIG. 2. A weight loss of 2.0% up to 150 °C was observed on the TGA curve. A sharp endotherm at about 130.7 °C (onset) was observed on the DSC curve. Polarized light microscopy (PLM) imaging of Form A indicated mainly irregular particles with size < 20 pm. Based on the small TGA weight loss and sharp DSC signal, Form A is an anhydrate.

[0105] DVS was measured via a SMS (Surface Measurement Systems) DVS Intrinsic. The relative humidity at 25 °C were calibrated against deliquescence point of LiCl, Mg(NO;h and KC1. Parameters for the DVS test are listed in Table 2-3. Table 2-3

[0106] DVS of Form A is shown in FIG. 3 and showed that water uptake increased from 0.04% to 0.33% in going from 70% RH to 95% RH.

[0107] Agilent 1260 HPLC was utilized for HPLC analysis. Table 2-4 provides the HPLC conditions. HPLC confirmed the purity of Form A was 98.62% (area).

Table 2-4

Example 3. Compound I Form B

[0108] Compound I Form B was obtained via adding anti-solvent H O into a NMP solution of Compound I, followed by drying at RT under vacuum. XRPD and TGA/DSC results for Form B are displayed in FIG. 4 and FIG. 5 respectively. The Form B analysis was conducted on the same instrumentation as the analysis for Form A in Example 2.

[0109] TGA showed a weight loss of 2.2% up to 150 °C. DSC showed a weak endotherm at 119.4 °C (peak) and a sharp endothermic peak at 130.9 °C (peak). After heating Form B to 119 °C and cooling to RT, Form A was obtained. Therefore, the endotherm at 119.4 °C signals the conversion of Form B to Form A. Based on the TGA, Form B is likely not a solvate or a hydrate.

[0110] A characterization summary for Form A and Form B is provided in Table 3-1 below.

Table 3-1

Example 4. Crystals of Compound II

[0111] Compound II was observed in samples of Compound I in alcohol solvent systems that were heated to 50 °C during the polymorph screening.

[0112] Compound II solid form was prepared via slow cooling of MeOH solution from 50 °C to 5 °C XRPD pattern is shown in Figure 6, which was consistent with Compound II. The XRPD analysis for Compound II was conducted on the same instrumentation as the analysis for Form A in Example 3. A weight loss of 1.7% up to 250 °C was observed on the TGA curve (FIG. 7, top line). On the DSC curve (FIG. 7, bottom line), a weak endotherm at 252.8 °C (peak) and a sharp endotherm at 259.7 °C (onset) were observed.

Example 5. Stability of Form A and Form B

[0113] The inter-conversion relationship between Form A and Form B was determined by slurry competition experiments in H O or MTBE at RT and 50 °C. Prior to the study, a Form A sample was used to saturate the corresponding solvent at the desired temperatures before filtration to obtain a near- saturated solution. Equal amounts (about 5 mg each) of Form A and Form B samples were weighed and then added in 1 mL of the prepared solution to form a suspension, followed by magnetically stirring (-1000 rpm) at RT and at 50 °C for about 2 days. Form A was obtained under these conditions, indicating that Form A was thermodynamically more stable than Form B in the temperature range of RT to 50 °C. Table 5-1 shows the results from this experiment.

Table 5-1

[0114] Based on the slurry competition results, Form A was further evaluated in tests for hygroscopicity, solid-state stability, kinetic solubility, stability in bio-relevant media and mechanical stability. Dynamic vapor sorption (DVS) data showed a water uptake of 0.04% at 70% RH, which increased to 0.33% at 95% RH.

[0115] Solid state stability of Form A was evaluated at 60 °C for one day, 25 °C and 60%RH and 40 °C and 75% RH for one week. No form change was observed for Form A after stability evaluation under these conditions. No change of HPFC purity was observed under 25 °C and 60% RH and under 40 °C and 75% RH after 1 week. A slight decrease in purity was observed after storage at 60 °C for 1 day.

[0116] Stability of Form A in simulated gastric fluid (SGF), fasted state simulated intestinal fluid (FaSSIF) and fed state simulated intestinal fluid (FeSSIF) at 37 °C was measured after 4 and 24 hrs. No form change was observed in all media after 4 and 24 hrs. The kinetic solubility of Form A in FaSSIF and FeSSIF was relatively low, and solubility in FeSSIF was higher than FaSSIF after 4 and 24 hrs.

Table 5-1 summarizes the results from solubility and stability testing in bio-relevant media.

Table 5-1

[0117] Mechanical stability of Form A was assessed by manual grinding, using an agate mortar and pestle. After grinding for 3 min, there was no form change, but some decrease in crystallinity was observed.

Example 6. Crystals of Compound I from ethanol

[0118] 20 mg Compound I was dissolved in 1 mL ethanol and kept in a half sealed 4 mL vial. The solution evaporated slowly at 4°C. Crystals were observed on the second day.

[0119] The crystal of Compound I was a colorless prism with the following dimensions:

0.30x0.10x0.10mm3. The symmetry of the crystal structure was assigned the triclinic space group P-1 with the following parameters: a = 8.2971(2) A, b = 10.7608(2) A, c = 14.5658(3) A, a = 70.529(2)°, b = 82.030(2)°, y = 71.545(2)°, V = 1162.26(5) A3, Z = 2, Dc = 1.482 Mg/m3, F(000) = 540.0, (Oi Ka) = 1.019 mm-1, and T = 100.0 (2) K.

X-ray crystallographic data

[0120] XRPD patterns were obtained on a Rigaku Oxford Diffraction XtaLAB Synergy four-circle diffractometer equipped with a HyPix-6000HE area detector:

Cryogenic system: Oxford Cryostream 800

Cu: l=1.54184 A, 50W, Micro focus source with multilayer mirror (m-CMF).

Distance from the crystal to the CCD detector: d = 35 mm Tube Voltage: 50 kV Tube Current: 1 m A

[0121] Standard XRPD patterns were collected using a Bruker D8 Advance diffractometer. The X- ray source is a Cu tube that was operated at 40 kV and 40 m A.

[0122] Fig. 8 shows the XRPD pattern for Compound I crystallized from ethanol and it corresponds to Compound I Form A. FIG. 10 shows an ORTEP plot for Compound I crystallized from ethanol.

TGA and DSC

[0123] The thermogravimetric analysis (TGA) was performed with a TA Q5000IR Thermal Analysis System with N2 as carrier gas. Differential scanning calorimeter analysis (DSC) was performed with a TA DSC Q2000 differential scanning calorimeter equipped with RCS cooling unit. The instrument parameters are listed below.

[0124] FIG. 9A shows the TGA analysis for Compound I crystallized from ethanol. FIG. 9B shows the DSC for Compound I crystallized from ethanol.

[0125] All patents and other references cited in the specification are indicative of the level of skill of those skilled in the art to which the disclosure pertains, and are incorporated by reference in their entireties, including any tables and figures, to the same extent as if each reference had been incorporated by reference in its entirety individually. [0126] One skilled in the art would readily appreciate that the present disclosure is well adapted to obtain the ends and advantages mentioned, as well as those inherent therein. The methods, variances, and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the disclosure. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the disclosure, are defined by the scope of the claims.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. A crystalline form of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH- pyrazol-4-yl)- 1 ,2,3 ,6-tetrahydro-7H-purine-7 -carboxylate (Compound I) :

wherein an X-ray powder diffraction pattern of Compound I comprises the following peaks: 15.23°±0.2°, 21.57°±0.2° and 22.43 °±0.2° (Compound I, Form A), expressed as 20 angles determined using Cu K-a (l = 1.5418 A) radiation.

2. The crystalline form of claim 1, wherein the diffraction pattern further comprises one or more peaks selected from 16.66°±0.2°, 18.79°±0.2°, and 23.40°±0.2°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation.

3. The crystalline form of claim 1, wherein the diffraction pattern further comprises two or more peaks selected from 6.26°±0.2°, 15.23°±0.2°, 16.66°±0.2°, 18.31°±0.2°, 18.79°±0.2°, 23.40°±0.2° and 25.18 °±0.2°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation.

4. The crystalline form of any one of claims 1-3, having an X-ray powder diffraction pattern substantially free of peaks at 18.15°±0.05°, and 22.76°±0.05°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation.

5. The crystalline form of any one of claims 1-4, wherein the diffraction pattern is substantially as shown in Figure 1.

6. The crystalline form of any one of claims 1-5, characterized by a differential scanning calorimetry (DSC) curve that shows an endotherm onset at about 131.9 °C.

7. The crystalline form of any one of claims 1-6, wherein the DSC curve is substantially as shown in Figure 2.

8. The crystalline form of any one of claims 1-7, wherein the dynamic vapor sorption (DVS) isotherm is substantially as shown in Figure 3.

9. The crystalline form of any one of claims 1-8, wherein the crystalline form is an anhydrate.

10. The crystalline form of any one of claims 1-9 prepared by addition of water to a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3- (trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7-carboxylate in acetone or tetrahydrofuran; or by addition of n-heptane to a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3- (trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7-carboxylate in dichloromethane or ethyl acetate.

11. The crystalline form of any one of claims 1-9 prepared by slow cooling or slow evaporation of a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6- tetrahydro-7H-purine-7 -carboxylate .

12. The crystalline form of any one of claims 1-9 prepared from a slurry of ethyl 3-ethyl-2,6-dioxo- l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7-carboxylate in a solvent, wherein the slurry is maintained at room temperature.

13. The crystalline form of claim 11 or claim 12, wherein the solvent is selected from ethanol, methyl-tert-butyl ether, dimethyl sulfoxide, dichloromethane, acetone, ethyl acetate, isopropyl acetate, toluene, isopropyl alcohol, methanol, tetrahydrofuran, n-heptane, water, or any combination thereof.

14. A crystalline form of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH- pyrazol-4-yl)- 1 ,2,3 ,6-tetrahydro-7H-purine-7 -carboxylate (Compound I) :

wherein an X-ray powder diffraction pattern of Compound I comprises the following peaks: 12.34°±0.2°, 18.15°±0.2°, and 22.76°±0.2°, (Compound I, Form B), expressed as 2Q angles determined using Cu K-a (l = 1.5418 A) radiation.

15. The crystalline form of claim 14, wherein the diffraction pattern further comprises one or more peaks selected from 15.56°±0.2°, and 21.04°±0.2°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation.

16. The crystalline form of claim 14, wherein the diffraction pattern further comprises two or more peaks expressed in ± 0.2 degrees 2-theta selected from 6.18 °±0.2°, 15.56°±0.2°, 21.04°±0.2°, and 22.76°±0.2°, expressed as 20 angles determined using Cu K-a (l = 1.5418A) radiation.

17. The crystalline form of any one of claims 14-16, having an X-ray powder diffraction pattern substantially free of peaks at 21.57°±0.05°, and 22.43°±0.05°.

18. The crystalline form of any one of claims 14-17, wherein the diffraction pattern is substantially as shown in Figure 4.

19. The crystalline form of any one of claims 14-18, characterized by a differential scanning calorimetry (DSC) curve that shows endotherm onsets at about 119.4 °C and at 130.9 °C.

20. The crystalline form of any one of claims 14-19, wherein the DSC curve is substantially as shown in Figure 5.

21. The crystalline form of any one of claims 14-20, wherein the crystalline form is an anhydrate.

22. The crystalline form of any one of claims 14-21 prepared by addition of water to a solution of ethyl 3-ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro- 7H-purine-7-carboxylate in N-methyl pyrrolidine.

23. The crystalline form of any one of claims 14-21 prepared by evaporation of a slurry of ethyl 3- ethyl-2,6-dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H- purine-7-carboxylate in a mixture of acetonitrile and isopropyl alcohol, wherein the slurry is maintained at a temperature ranging from - 20 °C to 5 °C.

24. The crystalline form of any one of claims 1-23, wherein the crystalline form is in a substantially pure form.

25. A pharmaceutical composition comprising the crystalline form of any one of claims 1 to 24, and one or more pharmaceutically acceptable carriers.

26. The pharmaceutical composition of claim 25, wherein at least 99% of the ethyl 3-ethyl-2,6- dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7- carboxylate is in a crystalline from.

27. The pharmaceutical composition of claim 25, wherein at least 95% of the ethyl 3-ethyl-2,6- dioxo-l-propyl-8-(l-(3-(trifluoromethyl)benzyl)-lH-pyrazol-4-yl)-l,2,3,6-tetrahydro-7H-purine-7- carboxylate is in a crystalline from.

28. A method for treating a disease or condition mediated, at least in part, by the A2_B adenosine receptor in a mammal, the method comprising administering to the mammal a therapeutically effective amount of the crystalline form of any one of claims 1-24, or the composition of any one of claims 25-27.

29. The method of claim 28, wherein the disease or condition is selected from the group consisting of cardiovascular diseases, fibrosis, neurological disorders, type I hypersensitivity disorders, chronic and acute liver diseases, lung diseases, renal diseases, diabetes, obesity, and cancer.

30. The method of claim 29, wherein the cancer is a hormone -related cancer.

31. The method of claim 30, wherein the hormone-related cancer is breast cancer, endometrial cancer, ovarian cancer, prostate cancer, testicular cancer, thyroid cancer or osteosarcoma.

32. The method of claim 30, wherein the hormone-related cancer is metastatic castration resistant prostate cancer.

33. The method of claim 30, wherein the hormone-related cancer is breast cancer.