Salts and Solid Forms of Compounds with APJ Receptor Activity

Abstract

The present disclosure relates to salts and solid forms of compounds that modulate APJ receptor activity and their use as therapeutic agents for treating diseases, disorders, or conditions associated with suppression or impairment of APJ receptor signaling, such as pulmonary arterial hypertension (PAH).

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to International Patent Application PCT / CN2022 / 131179, filed November 10, 2022, the entire contents of which are incorporated herein by reference.

[0002] The present disclosure relates generally to salts and solid forms of compounds that modulate APJ receptor activity, pharmaceutical compositions thereof, therapeutic uses thereof, and methods for their preparation. [Background technology]

[0003] The present disclosure relates to salts and solid forms of compounds that modulate apelin (APJ) receptor activity and their use as therapeutic agents for treating diseases, disorders, or conditions associated with repressed or impaired APJ receptor signaling, such as pulmonary arterial hypertension (PAH). Summary of the Invention

[0004] The present disclosure provides salts and solid forms of Compound I (CAS Registry Number: 2415197-87-2), as well as co-crystals and solvates thereof. Also described herein are methods for making the salts and solid forms of Compound I, pharmaceutical compositions containing the salts or solid forms of Compound I, and methods of using them in the treatment of diseases mediated by APJ receptor activity. [Brief explanation of the drawings]

[0005] [Figure 1A] Figure 1A shows the X-ray powder diffraction (XRPD) pattern of Compound I·choline salt Form A.

[0006] [Figure 1B] Figure 1B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I·choline salt Form A.

[0007] [Figure 2A] Figure 2A shows the X-ray powder diffraction (XRPD) of Compound I·choline salt Form B.

[0008] [Figure 2B] Figure 2B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I·choline salt form B.

[0009] [Figure 3A] FIG. 3A shows the X-ray powder diffraction (XRPD) of Compound I free acid Form A.

[0010] [Figure 3B] FIG. 3B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I free acid Form A.

[0011] [Figure 4A] FIG. 4A shows the X-ray powder diffraction (XRPD) of Compound I free acid Form B.

[0012] [Figure 4B] FIG. 4B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I free acid Form B.

[0013] [Figure 5A] FIG. 5A shows the X-ray powder diffraction (XRPD) of Compound I free acid Form C.

[0014] [Figure 5B] FIG. 5B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I free acid Form C.

[0015] [Figure 6A] FIG. 6A shows the X-ray powder diffraction (XRPD) of Compound I·sodium salt Form A.

[0016] [Figure 6B] Figure 6B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I·sodium salt Form A.

[0017] [Figure 7A] Figure 7A shows the X-ray powder diffraction (XRPD) of Compound I·potassium salt Form A.

[0018] [Figure 7B] Figure 7B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I·potassium salt Form A.

[0019] [Figure 8A] FIG. 8A shows the X-ray powder diffraction (XRPD) patterns of Compound I·potassium salt Form A and Compound I·potassium salt Form B.

[0020] [Figure 8B] Figure 8B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I·potassium salt Form B.

[0021] [Figure 9A] FIG. 9A shows the X-ray powder diffraction (XRPD) of Compound I calcium salt Form A.

[0022] [Figure 9B] Figure 9B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I calcium salt Form A.

[0023] [Figure 10A] FIG. 10A shows the X-ray powder diffraction (XRPD) of Compound I·magnesium salt Form A.

[0024] [Figure 10B] Figure 10B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I·magnesium salt Form A.

[0025] [Figure 11A]FIG. 11A shows an X-ray powder diffraction (XRPD) overlay of Compound I·sodium salt Form A (top) and Compound I·sodium salt Form B (bottom).

[0026] [Figure 11B] FIG. 11B shows the thermogravimetric analysis (TGA) / differential scanning calorimetry (DSC) curve of Compound I·sodium salt Form B.

[0027] [Figure 12] Figure 12 shows an ORTEP drawing of the crystal structure of Compound I·choline salt Form A.

[0028] Detailed Description The compound N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, designated herein as Compound I, has the formula: [ka] Compound I It has.

[0029] Compound I is a modulator of APJ receptor activity. Its synthesis and methods of use are described in PCT International Application Publication No. WO2020 / 073011, the entire contents of which are incorporated herein by reference.

[0030] 1.Definition As used herein, the following words and phrases are generally intended to have the meanings set forth below, unless the context in which they are used indicates otherwise.

[0031] The term "comprise" and variations thereof, such as "comprises" and "comprising," are to be construed in an open and inclusive sense, i.e., "including, but not limited to." Furthermore, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, reference to "a compound" includes a plurality of such compounds, and reference to "an assay" includes a reference to one or more assays and equivalents thereof known to those skilled in the art.

[0032] As used herein, reference to "about" a value or parameter includes (and describes) embodiments 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, the term "about x" includes the description "x".

[0033] Recitation of numerical ranges throughout this disclosure is intended to serve as a shorthand notation for referring individually to each separate value falling within the range, inclusive of the values ​​defining the range, and each separate value is included in the specification as if it were individually recited herein.

[0034] Provided herein is a form of Compound I or its salt, cocrystal or solvate. In one embodiment, a reference to a form of Compound I or its salt, cocrystal or solvate 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 its salt, cocrystal or solvate present in the composition is in the specified form. For example, in one embodiment, a reference to Compound I free acid 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 as a free acid is present in the composition as Form A.

[0035] The term "solid form" refers to a type of material in the solid state, including amorphous and crystalline forms. The term "crystalline form" refers to polymorphs and solvates (e.g., hydrates), etc. The term "polymorph" refers to a particular crystalline structure with specific physical properties, such as X-ray diffraction, melting point, etc.

[0036] The term "cocrystal" refers to a molecular complex in which a compound disclosed herein and one or more non-ionized co-crystal formers are bound via non-covalent interactions. In some embodiments, the co-crystals disclosed herein comprise a non-ionized form of compound I (e.g., a free form of compound I) and one or more non-ionized co-crystal formers, wherein the non-ionized compound I and the co-crystal formers are bound via non-covalent interactions. In some embodiments, the co-crystals disclosed herein comprise an ionized form of compound I (e.g., a salt of compound I) and one or more non-ionized co-crystal formers, wherein the ionized form of compound I and the co-crystal formers are bound via non-covalent interactions. The co-crystals may also exist in anhydrous or solvated forms. In certain embodiments, the co-crystals may have improved properties compared to the parent form (i.e., the free molecule, zwitterion, etc.) or a salt of the parent compound. Improved properties include increased solubility, increased dissolution, increased bioavailability, increased dose response, reduced hygroscopicity, increased stability, crystalline forms of compounds that are normally amorphous, crystalline forms of compounds that are difficult or impossible to salify, reduced form diversity, more desirable forms, etc. Methods for making and characterizing cocrystals are known to those skilled in the art.

[0037] The term "co-crystal former" or "co-former" means one or more pharmaceutically acceptable bases or pharmaceutically acceptable acids disclosed herein in association with Compound I or any other compound disclosed herein.

[0038] "Solvate" refers to a complex formed by the combination of solvent molecules with solute molecules or ions. The solvent can be an organic compound, an inorganic compound, or a mixture of both. As used herein, the term "solvate" includes "hydrates" (i.e., complexes formed by the combination of water molecules with solute molecules or ions), hemihydrates, channel hydrates, and the like. Some examples of solvents include, but are not limited to, methanol, N,N-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, and water. In general, solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present disclosure.

[0039] "Desolvated" means that solvent molecules have been partially or completely removed from a solvated form of Compound I described herein. Desolvation techniques for producing desolvated forms include, but are not limited to, exposing a Compound I form (solvate) to a vacuum, exposing the solvate to elevated temperatures, exposing the solvate to a stream of gas such as air or nitrogen, or any combination thereof. Thus, a desolvated Compound I form can be anhydrous, i.e., completely free of solvent molecules, or partially solvated, in which solvent molecules are present in stoichiometric or non-stoichiometric amounts.

[0040] The term "amorphous" refers to a state in which a material lacks long-range order at the molecular level and may exhibit the physical properties of a solid or a liquid, depending on temperature. Typically, such materials do not exhibit a distinctive X-ray diffraction pattern and exhibit the properties of a solid, but are more formally referred to as a liquid. Upon heating, a change from solid to liquid properties occurs, which is typically characterized by a second-order (glass transition) change of state.

[0041] The formulas or structures shown herein, including Compound I, are also intended to represent unlabeled and isotopically labeled forms of the compounds.It is understood that for any given atom, isotopes may be present essentially in proportions according to their natural occurrence, or one or more specific atoms may be enriched with one or more isotopes using synthetic methods known to those skilled in the art.Thus, hydrogen may be, for example, 1 H, 2 H, 3 H; carbon, for example, 11 C. 12 C. 13 C. 14 C; oxygen includes, for example, 16 O. 17 O. 18 O; nitrogen includes, for example, 13 N, 14 N, 15 Sulfur includes, for example, 32 S, 33 S, 34 S, 35 S, 36 S, 37 S, 38 S; fluoro includes, for example, 17 F, 18 F, 19 F; chloro, for example, 35 Cl, 36 Cl, 37 Cl, 38 Cl, 39 Contains Cl, etc.

[0042] As used herein, the terms "treat," "treating," "therapy," "therapies," and similar terms mean administering a substance, e.g., one or more solid, crystalline, or polymorphic forms of any of Compound I described herein, in an amount effective to prevent, alleviate, or ameliorate one or more symptoms, i.e., indications, of a disease or condition and / or prolong the survival of the subject being treated.

[0043] The term "administering" refers to oral administration, administration as a suppository, topical contact, intravenous administration, intraperitoneal administration, intramuscular administration, intralesional administration, intranasal administration, or subcutaneous administration, or implantation of a sustained-release device, such as a mini-osmotic pump, into a subject. Administration can be by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarteriolar, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial administration. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, and the like.

[0044] As used herein, the terms "modulating" or "modulate" refer to the effect of changing a biological activity, particularly a biological activity associated with a particular biomolecule, such as APJ receptor activity. For example, an agonist or antagonist of a particular biomolecule modulates the activity of the APJ receptor by increasing (e.g., agonist, activator) or decreasing (e.g., antagonist, inhibitor) the activity of that biomolecule. Such activity is typically measured by the inhibitory concentration (IC) of the compound relative to the inhibitor or activator, respectively. 50 ) or excitation concentration (EC 50 ) is shown.

[0045] As used herein, the term "composition" refers to a pharmaceutical formulation containing at least one pharmaceutically active compound (including any solid form) suitable for administration to a subject for therapeutic purposes. The composition may contain at least one pharmaceutically acceptable ingredient to provide an improved formulation of the compound, such as a suitable carrier or excipient.

[0046] The term "subject" or "patient" as used herein means a living organism that is treated with the compounds described herein, including, but not limited to, humans, other primates, sport animals, commercial animals such as cattle, farm animals such as horses, or pets such as dogs and cats.

[0047] The term "pharmaceutically acceptable" indicates that the indicated substance does not possess properties that would prevent a reasonably prudent physician from administering the substance to a patient, taking into account the disease or condition to be treated and the respective route of administration. For example, such substances are generally required to be essentially sterile, e.g., for injectables.

[0048] In this context, the term "therapeutically effective" or "effective amount" indicates that a substance or amount of a substance is effective to prevent, alleviate, or ameliorate one or more symptoms of a disease or medical condition, and / or prolong the survival of the subject being treated. A therapeutically effective amount will vary depending on the compound, the disorder or condition and its severity, and the age, weight, etc., of the mammal being treated. For example, an effective amount is an amount sufficient to produce a beneficial or desired clinical result. An effective amount can be provided all at once in a single administration, or in divided amounts that provide an effective amount in multiple administrations. The precise determination of what is considered an effective amount can be based on factors specific to each subject, such as the subject's size, age, injury and / or disease or injury being treated, and the length of time since the injury occurred or the disease began. One of ordinary skill 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.

[0049] In some embodiments, the phrase "substantially as shown in the figures" as applied to X-ray powder diffraction means including a variation of ±0.2° 2θ or ±0.1° 2θ, the phrase "substantially as shown in the figures" as applied to DSC thermograms means including a variation of ±3°C, and the phrase "substantially as shown in the figures" as applied to thermogravimetric analysis (TGA) means including a variation of ±2% weight loss.

[0050] "Substantially pure form (of a polymorph)" in some embodiments means that, for a referenced substance, at least 99.9% of the substance is the referenced polymorph. "Substantially pure form (of a polymorph)" in some embodiments means that, for a referenced substance, at least 99.5% of the substance is the referenced polymorph. "Substantially pure form (of a polymorph)" in some embodiments means that, for a referenced substance, at least 99% of the substance is the referenced polymorph. "Substantially pure form (of a polymorph)" in some embodiments means that, for a referenced substance, at least 98% of the substance is the referenced polymorph. "Substantially pure form (of a polymorph)" in some embodiments means that, for a referenced substance, at least 97% of the substance is the referenced polymorph. "Substantially pure form (of a polymorph)" in some embodiments means that, for a referenced substance, at least 96% of the substance is the referenced polymorph. "Substantially pure form (of a polymorph)" means, in some embodiments, that the referenced substance is at least 95% of the substance in the referenced polymorph. In the context of using, testing, or screening compounds that are or may be modulators, the term "contacting" means bringing the compound into sufficient proximity with a particular molecule, complex, cell, tissue, organism, or other particular substance so that potential binding interactions and / or chemical reactions between the compound and the other particular substance can occur.

[0051] 2. Salts and Forms of Compound I As noted above, the present disclosure provides salts and crystalline forms of the compound N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide (hereinafter "the Compound," "Compound I"), or a salt, co-crystal, or solvate thereof. Crystalline forms of Compound I, or a salt, co-crystal, or solvate thereof, and other forms of Compound I (e.g., amorphous forms), or a salt, co-crystal, or solvate thereof, are collectively referred to herein as "forms of Compound I."

[0052] In some embodiments, Compound I is a free acid. In some embodiments, Compound I is a salt. In some embodiments, Compound I is a pharmaceutically acceptable salt. In some embodiments, Compound I is a solvate. In some embodiments, Compound I is a hydrate. In some embodiments, Compound I is anhydrous.

[0053] In one embodiment, there is provided a solid form of N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide (Compound I), or a solvate thereof. In one embodiment, there is provided a solid form of a salt of N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide (Compound I), or a solvate thereof.

[0054] Salts of Compound I In one embodiment, the formula: [ka] [In the formula, X is sodium and n is 1; X is potassium and n is 1; X is calcium and n is 2; or X is magnesium and n is 2. or a solvate thereof.

[0055] In one embodiment, N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·sodium salt (Compound I·sodium salt) or a solvate thereof is provided. In one embodiment, N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·potassium salt (Compound I·potassium salt) or a solvate thereof is provided. In one embodiment, N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·calcium salt (Compound I·calcium salt) or a solvate thereof is provided. In one embodiment, there is provided N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide magnesium salt (Compound I magnesium salt) or a solvate thereof.

[0056] In one embodiment, the formula: [ka] wherein X is sodium and n is 1; X is potassium and n is 1; X is calcium and n is 2; or X is magnesium and n is 2. or a solvate thereof.

[0057] In one embodiment, crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·sodium salt (Compound I·sodium salt) or a solvate thereof is provided. In one embodiment, crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·potassium salt (Compound I·potassium salt) or a solvate thereof is provided. In one embodiment, crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·calcium salt (Compound I·calcium salt) or a solvate thereof is provided. In one embodiment, there is provided a crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide magnesium salt (Compound I magnesium salt) or a solvate thereof.

[0058] In one embodiment, the formula: [ka]

[0001] N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·choline salt (Compound I·choline salt) or a solvate thereof is provided, wherein the compound has the formula:

[0059] In some embodiments, the Compound I·choline salt is a hydrate. In some embodiments, the Compound I·choline salt is a monocholine salt (Compound I free acid:choline = about 1:1). In some embodiments, the Compound I·choline salt is a monocholine salt (Compound I free acid:choline = 1:0.9 to 1.1). In some embodiments, the Compound I·choline salt is a monocholine salt (Compound I free acid:choline = 1:0.9). In some embodiments, the Compound I·choline salt is a monocholine salt (Compound I free acid:choline = 1:1.1). In some embodiments, the Compound I·choline salt is a hydrate of the monocholine salt (Compound I free acid:choline = 1:1). In some embodiments, the hydrate is a non-stoichiometric hydrate (channel hydrate).

[0060] In some embodiments, the Compound I·choline salt or a solvate thereof is crystalline. In one embodiment, a crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·choline salt (Compound I·choline salt) or a solvate thereof is provided. In one embodiment, a hydrate of crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·choline salt (Compound I·choline salt) is provided. In some embodiments, the hydrate is a non-stoichiometric hydrate (channel hydrate).

[0061] Form of Compound I Compound I·Choline Salt Form A In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, choline salt Form A (Compound I, choline salt Form A). In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, choline salt Form A (Compound I, choline salt Form A), characterized by an X-ray powder diffraction pattern containing peaks at ±0.2 degrees two-theta selected from 12.9, 14.6, and 18.1, as measured with a diffractometer using Cu-Kα radiation.

[0062] In some embodiments, Compound I·choline salt Form A is further characterized by X-ray powder diffraction including one or more additional peaks at ±0.2 degrees 2θ selected from 9.9, 16.7, 17.5, 19.8, 22.7, and 23.1 as measured by a diffractometer using Cu-Kα radiation.

[0063] In some embodiments, Compound I·choline salt Form A is further characterized by an X-ray powder diffraction pattern substantially as depicted in FIG. 1A.

[0064] In some embodiments, Compound I·choline salt Form A is further characterized by a DSC comprising an endotherm at about 194-200°C (peak), or about 194-196°C (peak), or about 195-196°C (peak). In some embodiments, Compound I·choline salt Form A is further characterized by a DSC comprising an endotherm at about 194°C, or 194.2°C, or about 195°C, or 195.1°C, or 195.6°C, or about 196°C, or about 197°C, or about 200°C (peak). In some embodiments, Compound I·choline salt Form A is further characterized by a DSC comprising a broad endotherm at 104.0°C (peak) and a peak at 199.7°C (peak). In some embodiments, Compound I·choline salt Form A is further characterized by a DSC substantially as depicted in FIG. 1B.

[0065] In some embodiments, crystalline Compound I·choline salt Form A is prepared by slurrying Compound I free acid Form A and equimolar choline in acetone at room temperature for 4 days.

[0066] In some embodiments, Compound I·choline salt Form A is characterized by TGA showing a 1.9% weight loss up to 150° C. In some embodiments, Compound I·choline salt Form A is further characterized by TGA showing a 3.3% weight loss up to 150° C.

[0067] In some embodiments, the molar ratio of choline to Compound I free acid in Compound I·choline salt Form A is about 1.0. In some embodiments, the molar ratio of choline to Compound I free acid in Compound I·choline salt Form A is 0.9. In some embodiments, the molar ratio of choline to Compound I free acid in Compound I·choline salt Form A is between 1.1 and 0.9.

[0068] In some embodiments, Compound I choline salt Form A is a monocholine salt (Compound I free acid:choline = about 1:1). In some embodiments, Compound I choline salt Form A is a monocholine salt (Compound I free acid:choline = 1:0.9-1.1).

[0069] In some embodiments, the molar ratio of acetone / Compound I in Compound I·choline salt Form A is 0.04 (0.4 wt%). In some embodiments, the molar ratio of acetone / Compound I in Compound I·choline salt Form A is 0.01 (0.1 wt%). In some embodiments, the molar ratio of acetone / Compound I in Compound I·choline salt Form A is less than 0.01 (0.1 wt%).

[0070] In some embodiments, Compound I choline salt Form A is further characterized as a single crystal. Thus, in one embodiment, the unit cell parameters are: a = 8.4179(3) Å, b = 22.3688(7) Å, c = 14.7788(6) Å, α = 90°, β = 92.803(3)°, γ = 90°, V = 2779.49(17) Å. 3 Crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt Form A (Compound I choline salt Form A) is provided, having a molecular weight of 1.001, 1.002, 1.003, 1.004, 1.005, 1.006, 1.007, 1.008, 1.009 ...

[0071] Compound I·Choline Salt Form B In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, choline salt Form B (Compound I, choline salt Form B). In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, choline salt Form B (Compound I, choline salt Form B), characterized by an X-ray powder diffraction containing peaks at ±0.2 degrees two-theta selected from 11.0, 14.5, and 19.2, as measured with a diffractometer using Cu-Kα radiation.

[0072] In some embodiments, the compound I·choline salt Form B is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 18.4, 22.1, and 24.4 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, the compound I·choline salt Form B is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 18.7, 19.8, and 21.6 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, the compound I·choline salt Form B is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 18.4, 18.7, 19.8, 21.6, 22.1, and 24.4 as measured with a diffractometer using Cu-Kα radiation.

[0073] In some embodiments, Compound I·choline salt Form B is further characterized by an X-ray powder diffraction pattern substantially as depicted in FIG. 2A.

[0074] In some embodiments, Compound I·choline salt Form B is further characterized by a DSC comprising peaks at 74.4° C. (peak) and 172.1° C. (peak). In some embodiments, Compound I·choline salt Form B is further characterized by a DSC substantially as shown in FIG. 2B.

[0075] In some embodiments, Compound I·choline salt Form B is obtained by slurrying free acid Form A and equimolar choline in acetone and 1,4-dioxane, respectively, at room temperature for 4 days.

[0076] In some embodiments, Compound I·choline salt Form B is further characterized by TGA showing a weight loss of 5.9% up to 150°C.

[0077] In some embodiments, the molar ratio of choline to Compound I free acid in Compound I·choline salt Form B is 1.2. In some embodiments, the molar ratio of 1,4-dioxane to Compound I free acid in Compound I·choline salt Form B is 0.7 (6.2 wt %).

[0078] Compound I free acid form B In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide Form B (Compound I free acid Form B). In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide Form B (Compound I free acid Form B), characterized by an X-ray powder diffraction containing peaks at ±0.2 degrees two-theta selected from 9.5, 13.8, and 15.2, as measured on a diffractometer using Cu-Kα radiation.

[0079] In some embodiments, Compound I free acid Form B is further characterized by X-ray powder diffraction including one or more additional peaks at ±0.2 degrees 2θ selected from 14.2, 17.1, 19.6, 26.0, 26.9, and 27.9 degrees 2θ ±0.2 degrees 2θ as measured on a diffractometer using Cu-Kα radiation.

[0080] In some embodiments, Compound I free acid Form B is further characterized by an X-ray powder diffraction pattern substantially as depicted in Figure 4A.

[0081] In some embodiments, Compound I free acid Form B is further characterized by a DSC comprising two endotherms at 108.8° C. (peak) and 230.4° C. (peak). In some embodiments, crystalline Compound I free acid Form B is further characterized by a DSC substantially as shown in FIG. 4B.

[0082] In some embodiments, Compound I free acid Form B is obtained via slurrying Compound I free acid Form A in ACN:H 2 O (9:1, v / v) at room temperature for 4 days.

[0083] In some embodiments, Compound I free acid Form B is further characterized by TGA showing a weight loss of 8.5% up to 150°C.

[0084] In some embodiments, the molar ratio of ACN / Compound I free acid in Compound I free acid Form B is 0.8 (6.1 wt %).

[0085] Compound I sodium salt form A In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide sodium salt Form A (Compound I sodium salt Form A).

[0086] In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide sodium salt Form A (Compound I sodium salt Form A), characterized by an X-ray powder diffraction pattern containing peaks at ±0.2 degrees two-theta selected from 8.9, 15.5, and 26.3 as measured on a diffractometer using Cu-Kα radiation.

[0087] In some embodiments, Compound I·sodium salt Form A is further characterized by an X-ray powder diffraction pattern comprising one or more additional peaks at ±0.2 degrees 2θ selected from 12.2, 12.6, and 15.3 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·sodium salt Form A is further characterized by an X-ray powder diffraction pattern comprising one or more additional peaks at ±0.2 degrees 2θ selected from 7.8, 23.1, and 25.5 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·sodium salt Form A is further characterized by an X-ray powder diffraction pattern comprising one or more additional peaks at ±0.2 degrees 2θ selected from 7.8, 12.2, 12.6, 15.3, 23.1, and 25.5 as measured with a diffractometer using Cu-Kα radiation.

[0088] In some embodiments, Compound I sodium salt Form A is characterized by an X-ray powder diffraction pattern substantially as shown in Figure 6A.

[0089] In some embodiments, Compound I sodium salt Form A is further characterized by a DSC comprising two endotherms at 92.9 and 276.7° C. (peaks). In some embodiments, crystalline Compound I sodium salt Form A is further characterized by a DSC substantially as shown in FIG. 6B.

[0090] In some embodiments, Compound I sodium salt Form A is obtained via slurrying Compound I free acid Form A and equimolar NaOH in acetone at room temperature for 4 days.

[0091] In some embodiments, Compound I sodium salt Form A is further characterized by TGA showing a weight loss of 3.6% up to 150°C.

[0092] In some embodiments, Compound I sodium salt Form A is obtained via slurrying Compound I free acid Form A and equimolar NaOH in acetone at room temperature for 4 days.

[0093] In some embodiments, crystalline Compound I sodium salt Form A is a solvate. In some embodiments, crystalline Compound I sodium salt Form A is an acetone solvate. In some embodiments, the acetone solvate of Compound I sodium salt Form A has an acetone / Compound I molar ratio of 0.01 (0.1 wt %). In some embodiments, crystalline Compound I sodium salt Form A has a 1:1 molar ratio of sodium:Compound I free acid.

[0094] Compound I Potassium Salt Form A In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide potassium salt Form A (Compound I potassium salt Form A).

[0095] In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, potassium salt Form A (Compound I, potassium salt Form A), characterized by an X-ray powder diffraction pattern containing peaks at ±0.2 degrees two-theta selected from 12.3, 14.8, and 26.3 as measured on a diffractometer using Cu-Kα radiation.

[0096] In some embodiments, Compound I·potassium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 8.7, 22.3, and 25.9 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·potassium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 7.9, 15.7, and 21.3 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·potassium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 7.9, 8.7, 15.7, 21.3, 22.3, and 25.9 as measured with a diffractometer using Cu-Kα radiation.

[0097] In some embodiments, Compound I potassium salt Form A is characterized by an X-ray powder diffraction pattern substantially as shown in Figure 7A.

[0098] In some embodiments, Compound I potassium salt Form A is further characterized by a DSC comprising two endotherms at 74.2° C. and 304.6° C. (peaks). In some embodiments, Compound I potassium salt Form A is further characterized by a DSC substantially as shown in FIG. 7B.

[0099] In some embodiments, Compound I·potassium salt Form A is provided by slurrying Compound I free acid Form A and an equimolar amount of KOH in acetone at room temperature for 4 days.

[0100] In some embodiments, Compound I potassium salt Form A is further characterized by TGA showing a weight loss of 3.7% up to 150°C.

[0101] In some embodiments, the acetone solvate of Compound I potassium salt Form A has a molar ratio of acetone to Compound I of 0.03 (0.3 wt %). In some embodiments, Compound I potassium salt Form A has a molar ratio of potassium to Compound I free acid of 1:1.

[0102] Compound I Potassium Salt Form B In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, potassium salt Form B (Compound I, potassium salt Form B).

[0103] In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, potassium salt Form B (Compound I, potassium salt Form B), characterized by an X-ray powder diffraction pattern containing peaks at ±0.2 degrees two-theta selected from 6.8, 7.8, and 8.6, as measured on a diffractometer using Cu-Kα radiation.

[0104] In some embodiments, Compound I·potassium salt Form B is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 11.8, 14.7, and 22.3 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·potassium salt Form B is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 21.2, 25.8, and 26.3 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·potassium salt Form B is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 11.8, 14.7, 21.2, 22.3, 25.8, and 26.3 as measured with a diffractometer using Cu-Kα radiation.

[0105] In some embodiments, Compound I·potassium salt Form B is characterized by an X-ray powder diffraction pattern substantially as shown in FIG. 8A.

[0106] In some embodiments, Compound I·potassium salt Form B is further characterized by a DSC comprising an endotherm at 303.6° C. (peak). In some embodiments, Compound I·potassium salt Form B is further characterized by a DSC substantially as shown in FIG. 8B.

[0107] In some embodiments, Compound I·potassium salt Form B is provided by slurrying Compound I free acid Form A and equimolar KOH in MeOH at −20° C. for 4 days.

[0108] In some embodiments, Compound I·potassium salt Form B is further characterized by TGA showing a weight loss of 8.4% up to 150°C.

[0109] In some embodiments, Compound I potassium salt Form B is a non-solvate. In some embodiments, Compound I potassium salt Form B has a 1:1 potassium:Compound I free acid ratio.

[0110] Compound I calcium salt form A In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide calcium salt Form A (Compound I calcium salt Form A).

[0111] In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide calcium salt Form A (Compound I calcium salt Form A), characterized by an X-ray powder diffraction pattern containing peaks at ±0.2 degrees two-theta selected from 12.1, 16.6, and 25.6, as measured on a diffractometer using Cu-Kα radiation.

[0112] In some embodiments, Compound I·calcium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 20.4, 22.4, and 23.9 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·calcium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 6.7, 13.3, and 24.4 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, Compound I·calcium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 6.7, 13.3, 20.4, 22.4, 23.9, and 24.4 as measured with a diffractometer using Cu-Kα radiation.

[0113] In some embodiments, Compound I calcium salt Form A is characterized by an X-ray powder diffraction pattern substantially as shown in Figure 9A.

[0114] In some embodiments, Compound I-calcium salt Form A is further characterized by a DSC comprising two endotherms at 114.2 and 212.3° C. In some embodiments, Compound I-calcium salt Form A is further characterized by a DSC substantially as shown in FIG. 9B.

[0115] In some embodiments, Compound I calcium salt Form A is obtained by slurrying Compound I free acid Form A and equimolar Ca(OH) with ACN:HO (9:1, v / v) at room temperature for 4 days.

[0116] In some embodiments, Compound I calcium salt Form A is obtained by slurrying Compound I free acid Form A and Ca(OH) (base / acid molar ratio of 0.5) in ACN:HO (9:1, v / v) at room temperature for 1 day.

[0117] In some embodiments, Compound I calcium salt Form A is further characterized by TGA showing a two-stage weight loss of 2.1% up to 120°C and 6.8% from 120°C to 200°C.

[0118] In some embodiments, Compound I-calcium salt Form A has a molar ratio of ACN / Compound I of 0.07 (0.6 wt %). In some embodiments, Compound I-calcium salt Form A has a calcium:Compound I free acid ratio of 1:0.5.

[0119] Compound I Magnesium Salt Form A In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide magnesium salt Form A (Compound I magnesium salt Form A).

[0120] In one embodiment, there is provided crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide magnesium salt Form A (Compound I choline salt Form B), characterized by an X-ray powder diffraction pattern containing peaks at ±0.2 degrees two-theta selected from 13.7, 13.9, and 14.8 as measured on a diffractometer using Cu-Kα radiation.

[0121] In some embodiments, the compound I·magnesium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 19.0, 25.3, and 26.0 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, the compound I·magnesium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 22.0, 23.1, and 23.3 as measured with a diffractometer using Cu-Kα radiation. In some embodiments, the compound I·magnesium salt Form A is further characterized by an X-ray powder diffraction pattern containing one or more additional peaks at ±0.2 degrees 2θ selected from 19.0, 22.0, 23.1, 23.3, 25.3, and 26.0 as measured with a diffractometer using Cu-Kα radiation.

[0122] In some embodiments, Compound I·magnesium salt Form A is characterized by an X-ray powder diffraction pattern substantially as shown in FIG. 10A.

[0123] In some embodiments, Compound I·magnesium salt Form A is further characterized by a DSC comprising an endotherm at 136.6° C. (peak). In some embodiments, Compound I·magnesium salt Form A is further characterized by a DSC substantially as shown in FIG. 10B.

[0124] In some embodiments, Compound I·magnesium salt Form A is obtained by slurrying Compound I free acid Form A and equimolar Mg(OH) in ACN:HO (9:1, v / v) at room temperature for 4 days.

[0125] In some embodiments, Compound I·magnesium salt Form A is obtained by slurrying free acid Form A and Mg(OH) (molar ratio of Mg(OH) / Compound I free acid is 0.5) in ACN:HO (9:1, v / v) at room temperature for 2 days.

[0126] In some embodiments, Compound I·magnesium salt Form A is further characterized by TGA showing a weight loss of 12.0% up to 120°C.

[0127] In some embodiments, Compound I·magnesium salt Form A has a molar ratio of ACN / Compound I of 0.71 (5.7 wt %). In some embodiments, Compound I·magnesium salt Form A has a magnesium:Compound I free acid ratio of 1:0.5.

[0128] composition In some embodiments, compositions are provided comprising a salt or crystalline form of N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide (Compound I), or a salt or solvate thereof, as described herein.

[0129] In one embodiment, a composition is provided comprising a salt or crystalline form of N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide (Compound I), or a salt or solvate thereof, wherein 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%, at least 96%, or at least 99%) of Compound I present in the composition is the specified salt, crystalline form, or crystalline salt form.

[0130] In one embodiment, a composition is provided comprising N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·choline salt (Compound I·choline salt) or a solvate thereof, wherein 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%, at least 96%, or at least 99%) of the Compound I present in the composition is Compound I·choline salt.

[0131] In one embodiment, provided is a composition comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·choline salt Form A (Compound I·choline salt Form A) or a solvate thereof, wherein 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%, at least 96%, or at least 99%) of the Compound I present in the composition is Compound I·choline salt Form A.

[0132] In one embodiment, a composition is provided comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·choline salt Form B (Compound I·choline salt Form B) or a solvate thereof, wherein 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 present in the composition is Compound I·choline salt Form B.

[0133] In one embodiment, a composition is provided comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt Form B (Compound I free acid Form B) or a solvate thereof, wherein 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 present in the composition is Compound I free acid Form B.

[0134] In one embodiment, provided is a composition comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, sodium salt Form A (Compound I, sodium salt Form A) or a solvate thereof, wherein 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 the (Compound I) present in the composition is Compound I, sodium salt Form A.

[0135] In one embodiment, there is provided a composition comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·potassium salt Form A (Compound I·potassium salt Form A) or a solvate thereof, wherein 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 the (Compound I) present in the composition is (Compound I)·potassium salt Form A.

[0136] In one embodiment, a composition is provided comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·potassium salt Form B (Compound I·potassium salt Form B) or a solvate thereof, wherein 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 the Compound I present in the composition is (Compound I)·potassium salt Form B.

[0137] In one embodiment, provided is a composition comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·calcium salt Form A (Compound I·calcium salt Form A) or a solvate thereof, wherein 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) present in the composition is (Compound I)·calcium salt Form A.

[0138] In one embodiment, there is provided a composition comprising crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·magnesium salt Form A (Compound I·magnesium salt Form A) or a solvate thereof, wherein 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) present in the composition is (Compound I)·magnesium salt Form A.

[0139] In some embodiments, the composition is a pharmaceutical composition further comprising a pharmaceutically acceptable excipient.

[0140] In some embodiments, provided is a method for preparing crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·choline salt (Compound I·choline salt), comprising contacting Compound I or a salt thereof with choline in a solvent for a time sufficient to provide crystalline Compound I·choline salt.

[0141] In some embodiments, the solvent is acetone, hi some embodiments, the solvent is 1,4-dioxane.

[0142] In some embodiments, the contacting comprises adding an equimolar amount of choline to Compound I. In some embodiments, the contacting comprises adding an equimolar amount of choline to Compound I at a temperature of about 0°C to about 50°C. In some embodiments, the contacting comprises adding an equimolar amount of choline to Compound I at a temperature of about 20°C to about 30°C.

[0143] In some embodiments, the method further comprises isolating the crystalline Compound I·choline salt after the contacting step.

[0144] In some embodiments, the isolation comprises filtering, washing, and drying the crystalline (Compound I)·choline salt.

[0145] In some embodiments, a method of preparing a crystalline Compound I·choline salt is provided, wherein at least about 95% of the crystalline Compound I·choline salt is Form A.

[0146] In some embodiments, a method of preparing a crystalline Compound I·choline salt is provided, wherein at least about 95% of the crystalline Compound I·choline salt is Form B.

[0147] In some embodiments, a method for preparing crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide, sodium salt (Compound I, sodium salt) is provided, comprising contacting Compound I or a salt thereof with sodium hydroxide in a solvent for a time sufficient to provide crystalline Compound I, sodium salt.

[0148] In some embodiments, the solvent is acetone.

[0149] In some embodiments, the contacting comprises adding an equimolar amount of sodium hydroxide to Compound I. In some embodiments, the contacting comprises adding an equimolar amount of sodium hydroxide to Compound I at a temperature of about 0°C to about 50°C. In some embodiments, the contacting comprises adding an equimolar amount of sodium hydroxide to Compound I at a temperature of about 20°C to about 30°C.

[0150] In some embodiments, the method further comprises isolating the crystalline Compound I·sodium salt after the contacting step.

[0151] In some embodiments, the isolating comprises filtering, washing, and drying the crystalline Compound I·sodium salt.

[0152] In some embodiments, a method for preparing crystalline Compound I·sodium salt is provided, wherein at least about 95% of the crystalline Compound I·sodium salt is Form A.

[0153] In embodiments, provided is a method for preparing crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide·potassium salt (Compound I·potassium salt), comprising contacting Compound I or a salt thereof with potassium hydroxide in a solvent for a time sufficient to provide crystalline Compound I·potassium salt.

[0154] In some embodiments, the solvent is acetone.

[0155] In some embodiments, the contacting comprises adding an equimolar amount of potassium hydroxide to Compound I. In some embodiments, the contacting comprises adding an equimolar amount of potassium hydroxide to Compound I at a temperature of about 0°C to about 50°C. In some embodiments, the contacting comprises adding an equimolar amount of potassium hydroxide to Compound I at a temperature of about 20°C to about 30°C.

[0156] In some embodiments, the method further comprises isolating the crystalline Compound I·potassium salt after the contacting step.

[0157] In some embodiments, the isolating comprises filtering, washing, and drying the crystalline Compound I·potassium salt.

[0158] In some embodiments, a method for preparing crystalline Compound I·potassium salt is provided, wherein at least about 95% of the crystalline Compound I·potassium salt is Form A.

[0159] Pharmaceutical Compositions and Administration In some embodiments, a chemical entity that modulates (e.g., stimulates) the APJ receptor (e.g., a salt or crystalline form of N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide (Compound I), or a salt or solvate thereof, as described herein) is administered as a pharmaceutical composition comprising the chemical entity, one or more pharmaceutically acceptable excipients, and, optionally, one or more additional therapeutic agents as described herein.

[0160] In some embodiments, the chemical substance can be administered in combination with traditional pharmaceutical excipients.Pharmaceutically acceptable excipients include but are not limited to ion exchanger, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery system (SEDDS), such as d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms, such as Tween, poloxamer or other similar polymer delivery matrix, serum protein, such as human serum albumin, buffer substances, such as phosphate, Tris, glycine, sorbic acid, potassium sorbate, partial glyceride mixture of saturated vegetable fatty acids, water, salt or electrolyte, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylate, wax, polyethylene-polyoxypropylene-block polymer and wool fat. Chemically modified derivatives of cyclodextrins, such as α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin, or hydroxyalkyl cyclodextrins, including 2-hydroxypropyl-β-cyclodextrin and 3-hydroxypropyl-β-cyclodextrin, or other solubilized derivatives, can also be used to enhance delivery of the compounds described herein. Dosage forms or compositions can be prepared containing 0.005% to 100% of the chemicals described herein, with the remainder consisting of non-toxic excipients. Contemplated compositions can contain 0.001% to 100%, in one embodiment 0.1 to 95%, in another embodiment 75 to 85%, and in a further embodiment 20 to 80%, of the chemicals provided herein. Actual methods for preparing such dosage forms will be known or apparent to those skilled in the art. See, for example, Remington: The Science and Practice of Pharmacy, 22 nd Edition (Pharmaceutical Press, London, UK. 2012).

[0161] Route of administration and composition In some embodiments, the chemical entity described herein or pharmaceutical composition thereof may be administered to a subject in need thereof by any acceptable route of administration, including buccal, dermal, intracervical, endosinusial, intratracheal, enteral, epidural, intrainterstitial, intraperitoneal, intraarterial, intrabronchial, intrabursal, intracerebral, intracisternal, intraarterial, intradermal, intraductal, intraduodenal, intradural, intraepidermal, intraesophageal, intragastric, intragingival, intraileal, intralymphatic, intramedullary, intrathecal, intramuscular, and intraovarian. These administration methods include, but are not limited to, intraperitoneal, intraprostatic, pulmonary, intrasinus, intrathecal, intrasynovial, intratesticular, intrathecal, intraductal, intratumoral, intrauterine, intravascular, intravenous, nasal, nasogastric, oral, parenteral, transdermal, peridural, rectal, respiratory (inhalation), subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, tracheal, ureteral, urethral, ​​and vaginal administration.

[0162] Composition can be formulated for parenteral administration, for example, can be formulated for injection via intravenous, intramuscular, subcutaneous or intraperitoneal route.Typically, such composition can be prepared as an injection in the form of liquid solution or suspension, and can also be prepared in solid form suitable for preparing solution or suspension by adding liquid before injection, and preparation can also be emulsified.The preparation of such preparation will be known to those skilled in the art in light of the present disclosure.

[0163] Pharmaceutical forms suitable for injection include sterile aqueous solutions or dispersions; formulations containing sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the rapid preparation of sterile solutions or dispersions for injection. Generally, the form must be sterile and fluid enough to be easily syringable. It must also be stable under the conditions of manufacture and storage, and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.

[0164] The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it is preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0165] Sterile injectable solution can be prepared by incorporating the active compound in the required amount in suitable solvent with various other ingredients listed above as needed, and then sterilized by filtration.Generally, dispersion is prepared by incorporating various sterile active ingredients into a sterile vehicle that contains basic dispersion medium and other ingredients listed above as needed.For the preparation of sterile injectable solution, the preferred method of preparing sterile powder is vacuum drying and freeze-drying technology, and the powder of active ingredient plus any additional desired ingredients can be obtained from the solution that has been previously sterile filtered.

[0166] Pharmaceutically acceptable excipients that can be used in rectal compositions as gels, creams, enemas, or rectal suppositories include cocoa butter glycerides, synthetic polymers such as polyvinylpyrrolidone, PEG (such as PEG ointments), glycerin, glycerinated gelatin, hydrogenated vegetable oils, poloxamer, mixtures of fatty acid esters of polyethylene glycols of various molecular weights and polyethylene glycol petrolatum, anhydrous lanolin, shark liver oil, sodium saccharinate, menthol, sweet almond oil, sorbitol, sodium benzoate, anoxide SBN (anoxide SBN), and the like. SBN), vanilla essential oil, aerosol, parabens in phenoxyethanol, sodium methyl p-oxybenzoate, sodium propyl p-oxybenzoate, diethylamine, carbomer s, carbopol, methyloxybenzoate, macrogol cetostearyl ether, cocoyl caprylocaprate, isopropyl alcohol, propylene glycol, liquid paraffin, xanthan gum, carboxy-metabisulfite, disodium edetate, sodium benzoate, potassium metabisulfite, grapefruit seed extract, methylsulfonylmethane (MSM), lactic acid, glycine, vitamins such as vitamins A and E, and potassium acetate.

[0167] In certain embodiments, suppositories can be prepared by mixing the chemical substance described herein with suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol or suppository wax.These excipients or carriers are solid at ambient temperature but liquid at body temperature, and therefore melt in the rectum and release active compound.In other embodiments, the composition for rectal administration is in the form of an enema.

[0168] In other embodiments, the compounds described herein or pharmaceutical compositions thereof are suitable for local delivery to the digestive or GI tract by oral administration (eg, in solid or liquid dosage form).

[0169] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the chemical substance is mixed with one or more pharmaceutically acceptable excipients, such as sodium citrate or dicalcium phosphate, and / or: a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; c) humectants, such as glycerol; d) disintegrants, such as agar-agar, calcium carbonate, potato starch, or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retarding agents, such as paraffin; f) absorption enhancers, such as quaternary ammonium salts. The compound is mixed with g) wetting agents, such as cetyl alcohol and glycerol monostearate, h) absorbents, such as kaolin and bentonite clay, and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may contain buffering agents. Similar solid compositions can also be used as fillers in soft and hard-filled gelatin capsules, using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycols.

[0170] In one embodiment, the composition is in the form of a unit dosage form such as a pill or tablet, and thus the composition may contain, together with the chemical compound provided herein, diluents such as lactose, sucrose, dicalcium phosphate, etc.; lubricants such as magnesium stearate, etc.; and binders such as starch, acacia gum, polyvinylpyrrolidine, gelatin, cellulose, cellulose derivatives, etc. In another solid dosage form, powder, quince, solution or suspension (for example, in propylene carbonate, vegetable oil, PEG, poloxamer 124 or triglyceride) is encapsulated in a capsule (gelatin or cellulose-based capsule).A unit dosage form in which one or more chemical compounds provided herein or additional active agents are physically separated is also contemplated, and for example, a capsule containing granules of each agent (or a tablet in a capsule); a bilayer tablet; a two-compartment gel cap, etc., enteric-coated oral dosage form or delayed-release oral dosage form are also contemplated.

[0171] Other physiologically acceptable compounds include wetting agents, emulsifying agents, dispersing agents or preservatives which are particularly useful for preventing the growth or action of microorganisms. Various preservatives are well known, for example, phenol and ascorbic acid.

[0172] In certain embodiments, excipients are sterile and generally do not contain undesirable substances.These compositions can be sterilized by conventional well-known sterilization techniques.For various oral dosage form excipients such as tablets and capsules, sterility is not required.Usually, USP / NF standards are sufficient.

[0173] Ophthalmic compositions may include any one or more of the following, but are not limited to: viscogens (e.g., carboxymethylcellulose, glycerin, polyvinylpyrrolidone, polyethylene glycol); stabilizers (e.g., Pluronics (triblock copolymers), cyclodextrins); preservatives (e.g., benzalkonium chloride, ETDA, SofZia (boric acid, propylene glycol, sorbitol, and zinc chloride; Alcon Laboratories, Inc.), Purite (stabilized oxychloro complex; Allergan, Inc.)).

[0174] Topical compositions can include ointments and creams. Ointments are typically semi-solid preparations based on petrolatum or other petroleum derivatives. Creams containing selected active agents are typically viscous liquids or semi-solid emulsions, often either oil-in-water or water-in-oil. Cream bases are typically washable and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase, sometimes referred to as the "internal" phase, generally consists of petrolatum and a fatty alcohol (e.g., cetyl alcohol or stearyl alcohol). The aqueous phase usually, but not necessarily, exceeds the oil phase in volume and generally contains a moisturizer. Emulsifiers in cream formulations are generally nonionic, anionic, cationic, or amphoteric surfactants. Like other carriers or vehicles, ointment bases should be inert, stable, non-irritating, and non-sensitizing.

[0175] In any of the foregoing embodiments, the pharmaceutical compositions described herein can include one or more of lipids, interbilayer crosslinked multilamellar vesicles, biodegradable poly(D,L-lactic-co-glycolic acid) [PLGA]-based or polyanhydride-based nanoparticles or microparticles, and nanoporous particle-supported lipid bilayers.

[0176] Dosage Dosage can vary depending on the patient's needs, the severity of the condition being treated, and the specific compound used.Appropriate dosage for specific circumstances can be determined by those skilled in the art.In some cases, the total daily dosage can be divided and administered throughout the day or by means of providing continuous delivery.

[0177] In some embodiments, the compounds described herein are administered in a dose range of about 0.001 mg / Kg to about 500 mg / Kg (e.g., about 0.001 mg / Kg to about 200 mg / Kg; about 0.01 mg / Kg to about 200 mg / Kg; about 0.01 mg / Kg to about 150 mg / Kg; about 0.01 mg / Kg to about 100 mg / Kg; about 0.01 mg / Kg to about 50 mg / Kg; about 0.01 mg / Kg to about 10 mg / Kg; about 0.01 mg / Kg to about 5 mg / Kg; about 0.01 mg / Kg to about 1 mg / Kg). g / Kg; about 0.01mg / Kg to about 0.5mg / Kg; about 0.01mg / Kg to about 0.1mg / Kg; about 0.1mg / Kg to about 200mg / Kg; about 0.1mg / Kg to about 150mg / Kg; about 0.1mg / Kg to about 100mg / Kg; about 0.1mg / Kg to about 50mg / Kg; about 0.1mg / Kg to about 10mg / Kg; about 0.1mg / Kg to about 5mg / Kg; about 0.1mg / Kg to about 1mg / Kg; about 0.1mg / Kg to about 0.5mg / Kg).

[0178] Regimen The dosages can be administered daily (e.g., as a single dose or two or more divided doses) or non-daily (e.g., every other day, every third day, every third day, once a week, twice a week, once every two weeks, once a month).

[0179] In some embodiments, the administration period of the compounds described herein is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. In further embodiments, the period of time during which administration is stopped is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. In one embodiment, a therapeutic compound is administered to an individual for one period, followed by another period. In another embodiment, a therapeutic compound is administered for a first period and a second period following the first period, administration is stopped during the second period, followed by a third period during which administration of the therapeutic compound is initiated, and then a fourth period following the third period during which administration is stopped. In one aspect of this embodiment, the period of administration of a therapeutic compound followed by the period during which administration is stopped is repeated for a determined or undetermined period. In further embodiments, the duration of administration is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. In further embodiments, the period of time for which administration is suspended is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more.

[0180] Treatment method The present disclosure features methods of treating a subject (e.g., a human) having a disease, disorder, or condition in which decreased APJ receptor activity (e.g., suppression or impairment of APJ receptor signaling; e.g., suppression or impairment of apelin-APJ receptor signaling) or downregulation of endogenous apelin contributes to the pathology and / or symptoms and / or progression of the disease, disorder, or condition. In certain embodiments, the methods described herein can include, or can further include, treating one or more conditions associated with, coexisting with, or having sequelae of, any one or more of the conditions described herein.

[0181] In some embodiments, the method further comprises identifying the subject.In some embodiments, identifying comprises determining one or more of the following levels in the subject: leukotriene B4 level, pulmonary vascular resistance, pulmonary artery pressure, cardiac index, pulmonary capillary wedge pressure, right atrial pressure, 6-minute walk distance, brain natriuretic peptide level, atrial natriuretic peptide and spirometry.

[0182] In certain embodiments, the chemical compounds described herein modulate (e.g., decrease) pulmonary vascular resistance, modulate (e.g., decrease) right ventricular afterload, and modulate (e.g., decrease) mean pulmonary artery pressure. In certain embodiments, the chemical compounds described herein reduce the risk of right heart failure.

[0183] In certain embodiments, the chemical entities described herein regulate vascular tone, regulate fluid homeostasis, regulate renal function, regulate energy metabolism, regulate inflammatory responses, and regulate thrombosis.

[0184] Indications Pulmonary hypertension In some embodiments, the condition, disease or disorder is pulmonary arterial hypertension (PAH).Non-limiting examples of PAH and related conditions include idiopathic PAH, hereditary PAH (for example, BMPR2 mutation and other mutation), drug-induced or toxin-induced PAH, and the PAH associated with connective tissue disease (CTD) (for example, scleroderma, systemic lupus erythematosus, systemic sclerosis, Hashimoto's thyroiditis, Sjogren's syndrome and antiphospholipid antibody syndrome), HIV infection, portal hypertension, congenital heart disease and schistosomiasis.

[0185] In some embodiments, the PAH is idiopathic.

[0186] In other embodiments, the PAH is hereditary PAH, toxin- or drug-induced PAH; or PAH associated with one or more of the following: congenital heart disease, connective tissue disorders (e.g., scleroderma, systemic lupus erythematosus, systemic sclerosis, Hashimoto's thyroiditis, Sjogren's syndrome, and antiphospholipid syndrome), portal hypertension, BMPR2 mutations, schistosomiasis, and HIV infection.

[0187] In some embodiments, the pathological condition, disease or disorder is pulmonary hypertension other than PAH.Examples of such pathological conditions include but are not limited to the pulmonary hypertension caused by left heart disease (for example, left ventricular systolic dysfunction, left ventricular diastolic dysfunction, valvular heart disease and congenital / acquired left ventricular inflow / outflow obstruction and congenital cardiomyopathy), pulmonary disease and / or hypoxia (for example, chronic obstructive pulmonary disease, interstitial lung disease, other mixed restrictive and obstructive pulmonary diseases, sleep-disordered breathing, alveolar hypoventilation disorder, chronic exposure to high altitude, developmental pulmonary disease), pulmonary hypertension caused by chronic thromboembolic pulmonary hypertension and other pulmonary artery obstruction (for example, chronic thromboembolic pulmonary hypertension, other pulmonary artery obstruction), and pulmonary hypertension with unclear multifactorial mechanism (for example, blood disease, systemic disease, metabolic disease, etc.).

[0188] cardiovascular conditions, diseases or disorders In some embodiments, the pathology, disease or disorder is cardiovascular pathology, disease or disorder.Non-limiting examples of cardiovascular pathology, disease or disorder include coronary heart disease, acute coronary syndrome, peripheral vascular disease, angina pectoris, stroke, cerebrovascular accident, transient ischemic attack, heart failure, cardiomyopathy, myocardial infarction, myocardial remodeling after cardiac surgery, valvular heart disease, hypertension (for example, systemic hypertension, essential hypertension, pulmonary hypertension, portal hypertension, systolic hypertension), aortic aneurysm (for example, abdominal aortic aneurysm), atrial fibrillation, arrhythmia, atherosclerosis, Brugada syndrome, ischemic cardiovascular disease, peripheral arterial disease, preeclampsia, ventricular tachycardia and cardiac fibrosis.

[0189] In some embodiments, the cardiovascular condition, disease or disorder is heart failure.Non-limiting examples of heart failure include chronic heart failure, systolic heart failure, diastolic heart failure, diabetic heart failure, congestive heart failure, ejection fraction, heart failure with preserved ejection fraction, left ventricular dysfunction (for example, left ventricular dysfunction after myocardial infarction), right ventricular dysfunction, cardiac hypertrophy, myocardial remodeling and acute decompensated heart failure (ADHF).

[0190] In some embodiments, cardiovascular pathology, disease or disorder is pathology, disease or disorder that is associated with vascular pathology (for example, associated with increased vascular permeability and non-functional blood vessels).Non-limiting examples of such pathology, disease or disorder include vascular hypertrophy, vascular remodeling (for example, vascular hardening), atherosclerosis, peripheral arterial occlusive disease (PAOD), restenosis (for example, angioplasty restenosis), thrombosis and vascular permeability disorder, and ischemia and / or reperfusion injury (for example, ischemia and / or reperfusion injury of the heart, kidney and retina).In some embodiments, the pathology, disease or disorder is related to veins.Non-limiting examples of such pathology, disease or disorder include hemangioma, venous insufficiency, congestion or thrombosis.

[0191] In some embodiments, the chemical compounds described herein can improve myocardial contractility (e.g., cardiac relaxation), ventricular-arterial coupling, inotropic function, or lusitropic function in a subject suffering from a cardiovascular condition. In some embodiments, the chemical compounds described herein can increase the ejection fraction in a subject suffering from a cardiovascular condition.

[0192] Metabolic and homeostatic dysfunction and related conditions, diseases or disorders In some embodiments, the pathological condition, disease or disorder is related to metabolic dysfunction.Non-limiting examples of such pathological condition, disease or disorder include metabolic dysfunction, obesity, diabetes (for example, type II diabetes, gestational diabetes), diabetic complications (for example, metabolic syndrome, insulin resistance, organ damage caused by microvascular or macrovascular diseases, such as macroangiopathy and microangiopathy, diabetic neuropathy, diabetic retinopathy, cardiac autonomic neuropathy), kidney disease (for example, chronic kidney disease), edema, dyslipidemia, eating disorder, hyperphagia, bulimia, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, growth hormone disorder (for example, gigantism, acromegaly), galactorrhea and cardiac wasting.

[0193] In some embodiments, the condition, disease or disorder is associated with insufficient antidiuretic hormone (SIADH).Non-limiting examples of such condition, disease or disorder include neurogenic diabetes (for example, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy and other diabetic complications), lung cancer, septic shock and throat trouble.

[0194] In some embodiments, the pathological condition, disease, or disorder is associated with systemic inflammation. Non-limiting examples of such pathological conditions, diseases, or disorders include systemic inflammatory response syndrome (SIRs), sepsis (e.g., severe sepsis), and septic shock. In some embodiments, the pathological condition, disease, or disorder is associated with sepsis (e.g., complications, comorbidities, or sequelae of sepsis). Non-limiting examples of pathological conditions, diseases, or disorders associated with sepsis include sepsis-induced myocardial dysfunction, sepsis-related inflammatory response (e.g., systemic inflammation), sepsis-related hemodynamic changes, hypovolemia, sepsis-related organ failure (e.g., multiple organ failure, renal failure), acute kidney injury, vasoplegia, lung injury, abnormal vasopressin secretion, persistent hypertension associated with generalized vasodilation, refractory contractile reactivity, giant plasma capillary leak syndrome, coagulation / fibrinolysis imbalance, and metabolic disorders accentuated by elevated blood lactate levels. See, for example, Coquerel et al. Critical Care (2018) 22:10.

[0195] In some embodiments, the chemical entities described herein can modulate arginine vasopressin (AVP) or angiotensin receptors.

[0196] In some embodiments, the pathological condition, disease or disorder is related to the disturbance of body fluid homeostasis due to CNS-dependent and -independent effects.Non-limiting examples of such pathological condition, disease or disorder include renal failure (for example, acute and chronic renal failure), renal perfusion, renal insufficiency (for example, polycystic kidney disease), water diuresis and diuresis.

[0197] Dementia and related conditions, diseases or disorders In some embodiments, the condition, disease or disorder is dementia. Non-limiting examples of such conditions, diseases or disorders include senile dementia, vascular dementia, dementia due to genealogical degeneration diseases (e.g., Alzheimer's disease, Parkinson's disease, Pick's disease, Huntington's disease, etc.), dementia due to infectious diseases (e.g., slow-onset viral infections such as Creutzfeldt-Jakob disease), dementia associated with endocrine diseases, metabolic diseases or intoxication (e.g., hypothyroidism, vitamin B12 deficiency, alcoholism, various drug, metal or organic compound intoxication), dementia due to tumors (e.g., brain tumors), and dementia due to traumatic diseases (e.g., chronic subdural hematoma), depression, infantile hyperactivity syndrome (microencephalopathy), consciousness disorder, anxiety disorder, schizophrenia and phobia.

[0198] connective tissue disorders In some embodiments, the pathological condition, disease or disorder is connective tissue disorder.In certain embodiments, the connective tissue disorder is selected from the group consisting of scleroderma, systemic lupus erythematosus, systemic sclerosis, Hashimoto's thyroiditis, Sjogren's syndrome and antiphospholipid syndrome.In certain embodiments, the pathological condition, disease or disorder is systemic sclerosis.

[0199] fibrosis In some embodiments, the pathology, disease or disorder is fibrosis.In certain embodiments, the fibrosis is associated with organ or tissue selected from the group consisting of lung, liver, heart, mediastinum, bone marrow, retroperitoneum, skin, intestine, joint, reproductive organs and combinations thereof.In certain embodiments, the fibrosis is idiopathic pulmonary fibrosis (IPF).In certain embodiments, the fibrosis is hepatic fibrosis.In certain embodiments, the fibrosis is associated with non-alcoholic fatty liver disease (NAFLD).

[0200] Other conditions, diseases or disorders In some embodiments, the condition, disease, or disorder is liver disease. Non-limiting examples of such conditions, diseases, or disorders include alcoholic liver disease, toxicant-induced liver disease, viral-induced liver disease, and cirrhosis.

[0201] In some embodiments, said pathology, disease or disorder is pulmonary disease.Non-limiting examples of such pathology, disease or disorder include chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS) and amyotrophic lateral sclerosis.In some embodiments, said pathology, disease or disorder is retinal disease (for example, macular degeneration).

[0202] In some embodiments, the condition, disease or disorder is HIV infection, HIV neurodegeneration, neurodegenerative disease, cancer (e.g., breast cancer, lymphocytic leukemia, bladder cancer, ovarian cancer, prostate cancer, etc.), asthma, burns (e.g., sunburn), traumatic brain injury, pancreatitis, Turner syndrome, neuropathy, rheumatoid arthritis, spinal cord injury, immune function, inflammation, spinocerebellar degeneration, fractures, wounds, atopic dermatitis, osteoporosis, asthma, epilepsy and infertility.

[0203] Stem cell activation The chemical compounds described herein can also be used to activate stem cells (e.g., cardiac stem cells, such as endogenous cardiac stem cells). In some embodiments, the chemical compounds described herein can be used to regenerate tissue, aid in functional recovery after cell (e.g., bone marrow-derived mesenchymal stem cells) transplantation, increase cardiac stem cell proliferation (e.g., in patients (patients) who have suffered from myocardial infarction), reduce infarct size, promote cardiac repair, activate cells and progenitor cells in subjects after myocardial infarction, or reduce perfusion injury (e.g., during surgery, such as cardiac bypass surgery or heart transplantation).

[0204] Combination therapy The present disclosure contemplates both monotherapy and combination therapy regimens.

[0205] In some embodiments, the methods described herein may further include administering one or more additional therapies (e.g., one or more additional therapeutic agents and / or one or more treatment regimens) in combination with the administration of the compounds described herein. In some embodiments, the compounds described herein may be administered in combination with one or more additional therapeutic agents.

[0206] Representative additional therapeutic agents include, but are not limited to, agents for treating PAH, pulmonary hypertension, heart failure (e.g., ADHF, chronic heart failure), hypertension (e.g., systemic hypertension), amyotrophic lateral sclerosis, arrhythmias, asthma, atherosclerosis, atrial fibrillation, Brugada syndrome, burns (e.g., sunburn), cancer, cardiac fibrosis, cardiomyopathy, cerebrovascular accident, diabetes (e.g., gestational diabetes), septic shock, sepsis, renal failure, dyslipidemia, HIV neurodegeneration, inflammation, ischemic cardiovascular disease, liver disease, metabolic disorders, neurodegenerative diseases, obesity, peripheral arterial disease, preeclampsia, restenosis, transient ischemic attack, traumatic brain injury, ventricular tachycardia, edema, or immune function.

[0207] In some embodiments, the one or more additional therapeutic agents include, for example, those useful as therapeutic agents for PAH.Non-limiting examples include prostacyclin analogs (e.g., epoprostenol, treprostinil, iloprost), prostacyclin IP receptor (e.g., selexipag), endothelin receptor antagonists (e.g., bosentan, ambrisentan, macitentan), PDE5 inhibitors (e.g., sildenafil, tadalafil), soluble guanylate cyclase stimulators (e.g., riociguat), therapeutic agents for mitochondrial dysfunction (e.g., bardoxolone methyl), anti-inflammatory agents (e.g., rituximab, tocilizumab, ubenimex), and agents that regulate oxidative stress (e.g., dimethyl fumarate, intravenous iron).

[0208] In some embodiments, the one or more additional therapeutic agents include, for example, those useful as therapeutic agents for heart failure or hypertension. Non-limiting examples include α-blockers (e.g., doxazosin, prazosin, tamsulosin, terazosin), β-blockers (e.g., acebutolol, acetutolol, atenolol, bisoprol, bupranolol, carteolol, carvedilol, celiprolol, esmolol, mepindolol, metoprolol, nadolol, oxprenolol, penbutolol, pindolol, propranolol, taliprolol), calcium channel blockers (dihydropyridines (DHPs) (e.g., amlodipine, felodipine, isradipine, lacidipine, nicardipine, nifedipine, nigulpidine), and the like. diuretics (e.g., thiazide derivatives such as amiloride, chlorothiazide, hydrochlorthiazide, and methylchlorothiazide, including, but not limited to, thiazide derivatives such as thiazide derivatives, thiazide derivatives, and thiazide derivatives), centrally acting hypertensive drugs (e.g., thiazide derivatives, including, but not limited to, thiazide derivatives, such as thiazide derivatives, thiazide derivatives, and thiazide derivatives), diuretics ... and thiazide derivatives), diuretics (e.g., thiazide derivatives, including, but not limited to, thagents) (e.g., clonidine, guanabenz, guanfacine, methyldopa), angiotensin-converting enzyme (ACE) inhibitors (alaceptril, benazepril, benazaprilat, captopril, ceronapril, cilazapril, delapril, enalapril, analaprilat, fosinopril, lisinopril, moexipril, acetaminophen (e.g., acetaminophen, benzodiazepine, benzophenone, benzocaine, benzodiazepine ... Neuroreceptor blockers (ARBs) (e.g., candesartan, eprosartan, irbesartan, losartan, olmesartan, tasosartan, telmisartan, valsartan) and ARB / NEP dual inhibitors (e.g., a combination of valsartan and sacubitril), neutral endopeptidase (NEP) inhibitors (e.g., sacubitril), aldosterone synthase inhibitors (e.g., anastrozole, fadrozole, exemestane), endothelin antagonists (e.g., bosentan, enrasentan, atrasentan, darusentan, macitentan, sitaxentan, tezosentan), inhibitors of Fannie current (e.g., ivabradine), myosin activators (e.g., cardiac myosin activators), natriuretics agents), salt-excreting agents, vasodilators / vasorelaxants (e.g., nitrates), mineralocorticoid receptor antagonists, renin inhibitors, digitalis compounds, inotropes and β-receptor agonists, antihyperlipidemic agents, plasma HDL-raising agents, antihypercholesterolemic agents, cholesterol biosynthesis inhibitors (e.g., HMGCoA reductase inhibitors), LXR agonists, probucol, raloxifene, nicotinic acid, niacinamide, cholesterol absorption inhibitors, bile acid sequestrants (e.g., anion exchange resins, or quaternary amines, e.g., cholestyramine or colestipol), low-density lipoprotein receptor inducers, clofibrate, fenofibrate, bezafibrate, ciprofibrate, gemfibrizol, vitamins (e.g., vitamin B6, vitamin B 12 , antioxidant vitamins), platelet aggregation inhibitors, fibrinogen receptor antagonists, aspirin and fibric acid derivatives.

[0209] In some embodiments, the one or more additional therapeutic agents include, for example, those useful for treating diabetes. Non-limiting examples include sulfonylureas (e.g., chlorpropamide, tolbutamide, acetohexamide, tolazamide, glyburide, gliclazide, glinase, glimepiride, glipizide), biguanides (e.g., metformin), thiazolidinediones (e.g., ciglitazone, pioglitazone, troglitazone, rosiglitazone), insulin sensitizers (e.g., selective and non-selective activators of PPAR-alpha, PPAR-beta, and PPAR-gamma) related to the above, dehydroepiandrosterone (DHEA or its conjugates), and the like. monosulfate ester, also known as DHEA-SO4), antiglucocorticoids, TNF-alpha inhibitors, dipeptidyl peptidase IV (DPP4) inhibitors (e.g., sitagliptin, saxagliptin), GLP-1 agonists or analogs (e.g., exenatide), alpha-glucosidase inhibitors (e.g., acarbose, miglitol, and voglibose), pramlintide (a synthetic analog of the human hormone amylin), other insulin secretagogues (e.g., repaglinide, gliquidone, and nateglinide); and insulin.

[0210] In some embodiments, one or more additional therapeutic agents include, for example, those useful for treating obesity.Non-limiting examples include phenylpropanolamine, phentermine, diethylpropion, mazindol, fenfluramine, dexfenfluramine, phentiramine, beta3-adrenergic receptor agonists, sibutramine, gastrointestinal lipase inhibitors (e.g., orlistat), leptin, neuropeptide Y, enterostatin, cholecytokinin, bombesin, amylin, histamine H3 receptor, dopamine D2 receptor modulator, melanocyte-stimulating hormone, corticotropin-releasing factor, galanin and gamma-aminobutyric acid (GABA).

[0211] Other additional therapeutic agents include, but are not limited to, anti-atherosclerotic agents, anti-dyslipidemic agents, anti-hyperinsulinemic agents, anti-thrombotic agents, anti-retinopathic agents, anti-neuropathic agents, anti-nephrotic agents, anti-ischemic agents, anti-hyperlipidemic agents, anti-hypertriglyceridemic agents, anti-cholesterolemic agents, anti-restenosis agents, anti-pancreatic agents, appetite suppressants, memory enhancers, anti-dementia agents, cognition enhancers, appetite suppressants, agents for treating peripheral arterial disease, agents for treating malignancies, anti-inflammatory agents, aquaretics, digoxin, nitric oxide donors, hydralazine, ionotropes, vasopressin receptor antagonists, statins, anti-arrhythmic agents, phosphodiesterase inhibitors (e.g., PDE5 inhibitors), and nephroprotective agents.

[0212] In certain embodiments, the second therapeutic agent or regimen is administered to the subject prior to contact with or administration of the chemical agent (e.g., about 1 hour, or about 6 hours, or about 12 hours, or about 24 hours, or about 48 hours, or about 1 week, or about 1 month).

[0213] In other embodiments, the second therapeutic agent or regimen is administered to the subject at approximately the same time as contacting or administering the chemical substance. For example, the second therapeutic agent or regimen and the chemical substance are administered to the subject simultaneously in the same dosage form. For another example, the second therapeutic agent or regimen and the chemical substance are administered to the subject simultaneously in separate dosage forms.

[0214] In yet other embodiments, the second therapeutic agent or regimen is administered to the subject after contact with or administration of the chemical compound (e.g., about 1 hour, or about 6 hours, or about 12 hours, or about 24 hours, or about 48 hours, or about 1 week, or about 1 month). [Example]

[0215] Example 1. Synthesis of Compound I [ka] To a solution of 1 (3.4 kg, 8.27 mol, 1 eq) in DMSO (18.49 kg) was added MsNH (1.19 kg, 12.51 mol, 1.50 eq) and CsCO (8.06 kg, 24.74 mol, 3 eq) in four portions. The mixture was stirred at 120 °C for 20 h. LCMS showed that 1 was completely consumed. The mixture was cooled to 20-30 °C and then diluted with water (120 L). The mixture was filtered, and the filtrate was acidified to pH 3 with aqueous HCl (3 M). The suspension was filtered, and the filter cake was dissolved in THF (42.77 kg). After filtration, the solution was concentrated under reduced pressure to give the crude product. The crude product was purified by trituration from ACN (38.0 kg) at 90 °C for 16 hours to give compound I (1.93 kg), which was combined in 18 batches (obtained from 1 total of 9.86 kg) and recrystallized from THF:ACN = 1:5 (30 L) to give compound I (5.1 kg, 10.85 mol, 45% yield). HPLC (Waters XBridge C18 4.6 × 150 mm, 3.5 μm; oven temperature: 40 °C; flow rate: 1.0 mL / min; wavelength: 210 nm; mobile phase A: 0.1% H3PO4 in H2O, V / V; mobile phase B: ACN) and 1The material was confirmed by H NMR, with residual Pd at 4 ppm. 1 H NMR (400MHz, DMSO-d6):δ 11.05 (s, 1H), 8.31 (d, J = 2.4 Hz, 1H), 7.96 (d, J = 7.2 Hz, 1H), 7.86 (t, J = 8.0 Hz, 1H), 7.46 (t, J = 8.4 Hz, 1H), 6.85 (dd, J = 8.4, 13.6 Hz, 3H), 3.58 (s, 6H), 3.40 (dd, J = 7.2, 14.0 Hz, 2H), 3.21(s, 3H), 1.19 (t, J = 7.2 Hz, 3H).

[0216] Further analysis showed that Compound I prepared according to Example 1 was identified as Compound I free acid Form A.

[0217] Example 2: Synthesis of Compound I Choline Salt [ka] To a solution of compound I (2.03 kg, 4.31 mol, 1.00 eq) in THF (24.00 kg), the mixture was heated to reflux. Then, 47-50% choline (1.28 kg, 5.17 mol, 49% purity, 1.20 eq) was added dropwise at reflux and stirred for 4 h. The mixture was cooled to 15-20 °C and stirred for 16 h. The suspension was filtered, and the filter cake (combined from five separate batches) was triturated with EtOH (14.00 L) at 80 °C for 16 h. Next, after cooling to 15-20 °C, the suspension was filtered, and the filter cake was placed in a vacuum drying oven (45 °C, -0.092 MPa, 16 h) and then dried to obtain compound I choline salt.

[0218] 1H NMR (400MHz, DMSO-d6):δ 7.72-7.81 (m, 3H), 7.38 (t, J = 8.4 Hz, 1H), 6.80 (t, J = 4.4 Hz, 2H), 6.66 (dd, J = 0.8, 8.0 Hz, 1H), 5.35 (s, 1H), 3.81-3.86 (m, 2H), 3.56 (s, 6H), 3.33-3.42 (m, 4H), 3.11 (s, 9H), 2.71 (s, 3H), 1.01 (t, J = 7.2 Hz, 3H). LC-MS: Positive scan shows free form [M(C 21 H 21 N6O5S) +2H] + 471.1 and choline (C5H 14 NO) + 104.1 was detected; negative scans showed no free form of M(C 21 H 21 N6O5S) - 469.1 was detected.

[0219] Example 3: Salt Screening The starting material for the following salt screen was characterized by XRPD, TGA, and DSC. The XRPD pattern showed the material to be crystalline and was designated Free Acid Form A. The TGA / DSC results showed a 0.6% weight loss up to 150°C and an endotherm at 230.3°C (peak temperature). Due to the small weight loss in the TGA, Free Acid Form A was presumed to be an anhydrate.

[0220] The approximate solubility of the starting material (Compound I free acid form A) was measured in 14 solvents at room temperature. Approximately 2 mg of sample was placed in a 3 mL glass vial. The solvents in Table 1-3 were then added incrementally to the vial until the solid was visibly dissolved or a total volume of 2 mL was reached. The solubility results summarized in Table 1-1 were used to guide solvent selection in the screening experimental design. [Table 1]

[0221] Using Compound I free acid Form A as the starting material, a total of 32 salt screening experiments were performed using eight bases in four solvent systems. Approximately 15 mg of starting material and an equimolar amount of the corresponding salt former were added to 0.5 mL of solvent, followed by slurrying at room temperature for four days (clear samples were transferred to the slurry at -20 °C and subsequently evaporated at room temperature if still clear). The resulting suspension was centrifuged to recover the solid for XRPD testing, and the results are summarized in Tables 1-2. A total of seven crystalline salt hits and two new free acid polymorphs were obtained from the salt screening experiments, characterized by XRPD, TGA, DSC, and NMR or HPLC / IC. [Table 2]

[0222] Apparatus and method XRPD A PANalytical Empyrean and X' Pert3 X-ray powder diffractometer was used for the XRPD analysis. The XRPD parameters used are listed in Tables 1-3. [Table 3]

[0223] TGA and DSC TGA data were collected using a TA Instruments TA Q5000 / Discovery 5500 TGA. DSC was performed using a TA Instruments TA Q2000 / Discovery 2500 DSC. Detailed parameters used are listed in Tables 1-4. [Table 4]

[0224] DVS DVS was measured via the DVS Intrinsic of an SMS (Surface Measurement Systems). Relative humidity at 25°C was calibrated against the deliquescence points of LiCl, Mg(NO3)2, and KCl. DVS test parameters are listed in Tables 1-5. [Table 5]

[0225] PLM PLM images were taken with an Axio Lab. A1 upright microscope purchased from Carl Zeiss German.

[0226] Solution NMR Solution NMR was collected on a Bruker 400M NMR spectrometer using DMSO-d6.

[0227] HPLC / IC A Waters H-Class UPLC was used, and the detailed chromatographic conditions are listed in Tables 1-6. The IC parameters are listed in Tables 1-7. [Table 6] [Table 7]

[0228] Characterization of salt forms Seven salt forms were obtained from the salt screening and further experiments and were characterized by XRPD, TGA and DSC. The salt stoichiometry was determined by HPLC / IC or 1 All characterization results were determined using 1 H NMR and are summarized in Tables 1-8. [Table 8]

[0229] Sodium salt Compound I sodium salt Form A (Na salt Form A) was obtained by slurrying Compound I free acid Form A and an equimolar amount of NaOH in acetone at room temperature for 4 days. The XRPD pattern is shown in Figure 6A. The TGA / DSC curve of Compound I sodium salt Form A is shown in Figure 6B. The curve showed a 3.6% weight loss up to 150 °C and two endotherms at 92.9 °C and 276.7 °C (peaks). 1 H NMR indicated that the acetone / API molar ratio was 0.01 (0.1 wt%). HPLC / IC indicated that the molar ratio was 1.0 (base / FA).

[0230] K salt Compound I potassium salt Form A (K salt Form A) was obtained by slurrying the free acid Form A and an equimolar amount of KOH in acetone at room temperature for 4 days. Compound I potassium salt Form B (K salt Form B) was obtained by slurrying the free acid Form A and an equimolar amount of KOH in MeOH at -20 °C for 4 days. The XRPD patterns are shown in Figures 7A and 8A, respectively.

[0231] The TGA / DSC curve of compound I·potassium salt form A (K salt form A) is shown in Figure 7B. The curve showed a weight loss of 3.7% up to 150 °C and two endotherms at 74.2 °C and 304.6 °C (peaks). 1 H NMR indicated that the acetone / API molar ratio was 0.03 (0.3 wt%). HPLC / IC results indicated that the molar ratio was 1.0 (base / FA).

[0232] The TGA / DSC curve of compound I·potassium salt form B (K salt form B) is shown in Figure 8B. The curve showed a weight loss of 8.4% up to 150 °C and an endotherm at 303.6 °C (peak). 1 H NMR showed that no residual MeOH was detected. HPLC / IC showed the molar ratio to be 1.0 (base / FA).

[0233] Calcium salt Compound I calcium salt Form A (Ca salt Form A) was obtained by slurrying the free acid Form A and equimolar Ca(OH) in ACN:H2O (9:1, v / v) for 4 days at room temperature. XRPD patterns showed residual Ca(OH)2 in this sample.

[0234] Another compound I calcium salt form A (Ca salt form A) was obtained by slurrying the free acid form A and Ca(OH) (base / acid molar ratio 0.5) in ACN:HO (9:1, v / v) for 1 day at room temperature. The XRPD pattern is shown in Figure 9A. The TGA / DSC curve of compound I calcium salt form A (Ca salt form A) is shown in Figure 9B. The curve showed a two-step weight loss of 2.1% up to 120 °C and 6.8% from 120 °C to 200 °C, with two endotherms at 114.2 °C and 212.3 °C. 1 H NMR indicated that the molar ratio of ACN / API was 0.07 (0.6 wt%). HPLC / IC indicated that the molar ratio was 0.5 (base / FA).

[0235] Mg salt Compound I magnesium salt Form A (Mg salt Form A) was obtained by slurrying the free acid Form A and equimolar Mg(OH) in ACN:HO (9:1, v / v) at room temperature for 4 days. Residual Mg(OH) was detected in the XRPD pattern.

[0236] Compound I magnesium salt Form A (Mg salt Form A) was obtained by slurrying the free acid Form A and Mg(OH) (base / acid molar ratio 0.5) in ACN:HO (9:1, v / v) at room temperature for 2 days. The XRPD pattern is shown in Figure 10A. The TGA / DSC curve of Compound I magnesium salt Form A (Mg salt Form A) is shown in Figure 10B. The curve showed a 12.0% weight loss up to 120 °C and an endotherm at 136.6 °C (peak). 1 H NMR showed the ACN / API molar ratio to be 0.71 (5.7 wt%). HPLC / IC showed the molar ratio to be 0.5 (base / FA).

[0237] Choline salts Compound I·choline salt Form A and Form B were obtained by slurrying the free acid Form A and equimolar choline, respectively, in acetone and 1,4-dioxane at room temperature for 4 days.

[0238] The TGA / DSC curve for choline salt Form A showed a weight loss of 3.3% up to 150°C and an endotherm at 196.6°C (peak). 1 H NMR showed that the molar ratio of choline / Compound I free acid was 0.9 and the molar ratio of acetone / API was 0.01 (0.1 wt %).

[0239] The TGA / DSC curve of choline salt Form B showed a weight loss of 5.9% up to 150° C. and two endotherms at 74.4° C. and 172.1° C. (peaks). 1 H NMR indicated that the molar ratio of choline / Compound I free acid was 1.2 and the molar ratio of 1,4-dioxane / API was 0.7 (6.2 wt%).

[0240] Free acid Compound I free acid form B (free acid form B) was obtained by slurrying free acid form A in ACN:HO (9:1, v / v) at room temperature for 4 days, and the XRPD is shown in Figure 4A.

[0241] The TGA / DSC curve of Compound I free acid form B (free acid form B) is shown in Figure 4B. It showed a weight loss of 8.5% up to 150°C and two endotherms at 108.8°C and 230.4°C (peaks). 1 H NMR showed that the ACN / API molar ratio was 0.8 (6.1 wt%).

[0242] Compound I free acid form C (free acid form C) was obtained by heating free acid form B to 150° C. and cooling to room temperature, and the XRPD pattern is shown in FIG. 5A.

[0243] The TGA / DSC curve of Compound I free acid form C (free acid form C) is shown in Figure 5B. The curve showed a weight loss of 3.1% up to 150°C and an endotherm at 229.9°C (peak). 1 No residual ACN was detected by 1 H NMR.

[0244] Reconstitution of selected salts Considering the small weight loss in TGA and the sharp endotherm in DSC, Na salt Form A, K salt Form A, and choline salt Form A were selected for reconstitution on a 300 mg scale. All samples were successfully reconstituted and characterized by XRPD, TGA, DSC, NMR, or HPLC / IC, and are summarized in Tables 1-9. [Table 9]

[0245] Sodium salt Compound I sodium salt Form A (Na salt Form A) was reconstituted by slurrying approximately 300 mg of Compound I free acid Form A and an equimolar amount of NaOH in acetone at room temperature for 24 hours. The XRPD pattern is shown in Figure 6A. The TGA / DSC curve of Na salt Form A is shown in Figure 6B. The curve showed a 3.8% weight loss up to 150 °C and two endotherms at 89.2 °C and 278.1 °C (peaks). HPLC / IC results indicated a molar ratio of 1.0 (base / FA). Based on the weight loss by step TGA and the corresponding endotherm in DSC, Na salt Form A was presumed to be a hydrate.

[0246] K salt Compound I potassium salt Form A (K salt Form A) was reconstituted by slurrying approximately 300 mg of Compound I free acid Form A and an equimolar amount of KOH in acetone at room temperature for 48 hours. The XRPD pattern is shown in Figure 7A. The TGA / DSC curve of K salt Form A is shown in Figure 7B. The curve showed a 2.3% weight loss up to 150 °C and two endotherms at 74.2 °C and 306.1 °C (peaks). HPLC / IC results indicated a molar ratio of 0.9 (base / FA). Based on the weight loss by step TGA and the corresponding endotherm in DSC, K salt Form A was presumed to be a hydrate.

[0247] Choline salts Compound I·choline salt Form A (choline salt Form A) was reconstituted by slurrying approximately 300 mg of free acid Form A and an equimolar amount of choline in acetone at room temperature for 24 hours. The XRPD pattern is shown in Figure 1A. The TGA / DSC curve of choline salt Form A is shown in Figure 1B. The curve showed a 1.9% weight loss up to 150 °C and an endotherm at 196.6 °C (peak). 1 H NMR indicated that the base / FA molar ratio was 1.0 and the acetone / API molar ratio was 0.01 (0.1 wt%). Based on the small weight loss in TGA, Compound I·choline salt Form A was presumed to be anhydrous.

[0248] Salt Rating Three reconstituted salt samples were used for salt evaluation, including hygroscopicity, kinetic solubility, and solid-state stability. The starting material, Compound I free acid Form A, was also evaluated for comparison.

[0249] Hygroscopic To evaluate the moisture absorption of Compound I free acid Form A and the reformulated salt forms, DVS isotherm plots were collected at 25 °C from 0% RH to 95% RH. XRPD characterization was performed on the samples after DVS testing. The DVS evaluation results are summarized in Tables 1-10. Based on the results, Compound I free acid Form A exhibited the lowest moisture absorption. Compound I Na salt Form A transformed into a new form after DVS testing. There was no significant difference between Compound I K salt Form A and Compound I choline salt Form A. [Table 10]

[0250] kinetic solubility The kinetic solubilities of Compound I free acid Form A and the reconstituted salt forms were determined in water and three biologically relevant media.

[0251] Weigh out approximately 20 mg of material and add it to 4 mL of water, SGF, FaSSIF, or FeSSIF. 1 The salts were placed in a container and rolled at 25 rpm at 37 °C for 1, 2, 4, and 24 hours. At each time point, approximately 0.8 mL of the suspension was sampled, centrifuged, and filtered. The solids were examined by XRPD, and the filtrates by HPLC and pH. Based on the results, all salts exhibited similar solubility profiles, with the solubility of the salts in water and FaSSIF being higher than that of the free acid form A, which was presumably due to differences in pH. The XRPD results indicated that disproportionation was observed in all salt samples after the solubility tests.

[0252] solid state stability Compound I free acid Form A and the reconstituted salt forms were subjected to solid-state stability evaluation at 25°C / 60% RH and 40°C / 75% RH for one week. Physical and chemical stability were evaluated by XRPD and HPLC purity, respectively. Based on the results, no change in morphology or obvious decrease in HPLC purity was observed in any of the samples.

[0253] conclusion Starting with Compound I free acid Form A, salt screening was performed under 32 conditions using eight bases in four solvent systems. A total of seven crystalline salt hits and two novel free acid forms were obtained. Based on their characterization results (small weight loss in TGA and sharp endothermic signals on DSC curves), Compound I sodium salt Form A, Compound I potassium salt Form A, and Compound I choline salt Form A were selected for reconstitution, and all were successfully reconstituted.

[0254] The reformulated salts were used for salt evaluation, and Compound I free acid Form A was also evaluated for comparison. DVS results showed that free acid Form A exhibited the lowest water absorption, and morphological changes were observed in Compound I·Na salt Form A after DVS. Kinetic solubility results showed that all salts exhibited similar solubility profiles, with the solubility of the salts in HO and FaSSIF being higher than that of the free acid. Solid-state stability evaluation results showed that no morphological changes or obvious loss of HPLC purity were observed in any samples after 1 week of storage at 25°C / 60% RH or 40°C / 75% RH. Based on the salt evaluation and solid-state characterization results, Compound I·choline salt Form A was selected and reformulated at a 2 g scale for further PK studies.

[0255] Combining the results of salt evaluation and PK studies, compound I·choline salt was selected for further polymorph screening.

[0256] Reconstitution of choline salts Compound I·choline salt Form A was reprepared on a 2 g scale by the procedures listed below. The sample was characterized by XRPD, TGA, DSC, and NMR. The XRPD pattern is shown in Figure 1A. The TGA / DSC curve of Compound I·choline salt Form A is shown in Figure 1B. The curve showed a 2.9% weight loss up to 150 °C and an endotherm at 195.6 °C (peak). 1 H NMR showed that the base / FA molar ratio was 1.0 and the acetone / API molar ratio was 0.03 (0.3 wt%). 1. Weigh out 1.05 g of choline (49 wt%, 4.25 mmol) into a 100 mL reactor; 2. Add 30 mL of acetone to the reactor to obtain a suspension; 3. Weigh out 2.00 g free acid (4.25 mmol) into the reactor; 4. Stir at room temperature for 24 hours; 5. The suspension is filtered and the solid is dried under vacuum at room temperature (to give about 1.5 g of product).

[0257] Example 4: Polymorph Screening and Evaluation of Compound I Choline Salt The objective of this project was to perform polymorph screening and characterization of the choline salt and select a lead form for further study. The starting material was characterized by XRPD, TGA, and DSC. The TGA / DSC results showed a 0.5% weight loss up to 150 °C and an endotherm at 231.1 °C (peak temperature). Based on the small weight loss in TGA, the free acid form A was presumed to be an anhydrate.

[0258] Apparatus and method The XRPD, TGA, DSC, PLM, NMR and HPLC instruments and methods used were the same as in Example 3.

[0259] summary Fourteen slurry experiments were performed at 50° C. for each of the reconstituted choline salt Form A and free acid Form A. XRPD results indicated that only choline salt Form A and free acid Form A were obtained from the polymorph screening experiments.

[0260] The reconstituted choline salt Form A was used for evaluation of solubility, solid state stability, etc. The free acid Form A was selected as a comparison for pH solubility and solid state stability evaluations.

[0261] The pH solubility results showed that the choline salt form A had higher solubility than the free acid form A in pH buffer solutions.

[0262] Solid-state stability evaluation showed that choline salt Form A and free acid Form A did not change in morphology or show any significant decrease in HPLC purity after storage at 25°C / 60% RH or 40°C / 75% RH for 54 and 91 days, respectively.

[0263] The evaluation results showed that choline salt Form A was physically and chemically stable for 91 days at 25°C / 60% RH and 40°C / 75% RH and showed higher solubility than free acid Form A in pH buffer solutions.

[0264] Reconstitution of Choline Salt Form A Compound I choline salt Form A was reconstituted by slurrying 3.5 g of Compound I free acid Form A and an equimolar amount of choline in acetone at room temperature for 4 days. The XRPD pattern is shown in Figure 1A. The TGA / DSC curve of choline salt Form A is shown in Figure 1B. The curve showed a 1.9% weight loss up to 150 °C and an endotherm at 195.6 °C (peak). 1 H NMR analysis indicated that the base / FA molar ratio was 1.0 and the acetone / API molar ratio was 0.04 (0.4 wt%). PLM images showed that choline salt Form A was composed of irregular particles approximately 10 μm in size. Compound I·choline salt Form A was highly soluble in polar solvents such as methanol, DMSO, and water, but was less soluble in certain nonpolar solvents such as IPA, acetone, MEK, MIBK, ethyl acetate, isopropyl acetate, MTBE, THF, 2-MeTHF, CHCl3, n-heptane, toluene, DCM, and 1,4-dioxane.

[0265] Polymorphism Screening To preliminarily investigate the polymorphic risk of the choline salt and free acid, 14 slurry experiments were conducted at 50°C for choline salt Form A and free acid Form A, respectively. Approximately 15 mg of choline salt Form A was suspended in 0.5 mL of the corresponding solvent in an HPLC vial. After the suspension was magnetically stirred (approximately 750 rpm) at 50°C for approximately 3 days, the resulting solid form was choline salt Form A, and the results are summarized in Table 4-1. Approximately 15 mg of free acid Form A was suspended in 0.5 mL of the corresponding solvent in an HPLC vial. After the suspension was magnetically stirred (approximately 750 rpm) at 50°C for approximately 2 days, the resulting solid form was free acid Form A, and the results are summarized in Table 4-2. [Table 11] [Table 12]

[0266] Solution crystallization of Compound I·choline salt form A Compound I·choline salt Form A was reprepared by crystallization in THF and CHCl3 at room temperature on an approximately 100 mg scale. The procedure is listed in Table 4-3. XRPD showed that Compound I·choline salt Form A was successfully obtained by solution crystallization in THF and CHCl3. [Table 13]

[0267] conclusion Fourteen slurry experiments were performed at 50 °C for each of the reconstituted Compound I·choline salt Form A and free acid Form A. XRPD results indicated that only Compound I·choline salt Form A and free acid Form A were obtained from the polymorph screening experiments.

[0268] The reconstituted Compound I·choline salt Form A was used for evaluations such as solubility and solid-state stability. The free acid Form A was selected as a comparison for pH, solubility, and solid-state stability evaluations.

[0269] The pH solubility results showed that Compound I·choline salt Form A had higher solubility than the free acid Form A in pH buffer solutions.

[0270] Solid-state stability evaluation showed that Compound I·choline salt Form A and free acid Form A did not undergo any change in morphology or significant loss of HPLC purity after storage at 25°C / 60% RH or 40°C / 75% RH for 54 and 91 days, respectively.

[0271] The evaluation results showed that Compound I·choline salt Form A was physically and chemically stable for 91 days at 25°C / 60% RH and 40°C / 75% RH and showed higher solubility than the free acid Form A in pH buffer solutions.

[0272] Compound I choline salt Form A exhibited more desirable solid-state properties than the other salts / solid forms tested. For example, the sodium salt was hygroscopic and less stable after exposure to high humidity when compared to Compound I choline salt Form A. Compound I magnesium salt Form A and the calcium and potassium salts of Compound I are solvates, and Compound I choline salt Form A is believed to be a stable nonstoichiometric hydrate with a moderate melting point.

[0273] Example 5: Polymorph Screening and Single Crystal Structure Determination of Compound I Choline Salt This project included polymorph screening and single crystal studies of Compound I·choline salt Form A. The objective of the polymorph screening was to explore potential crystalline forms and determine whether Compound I·choline salt Form A is a suitable form for further pharmaceutical development. The objective of the single crystal study was to understand the crystalline form of Compound I·choline salt Form A and confirm whether it is a salt or a co-crystal.

[0274] The received starting materials were analyzed by powder X-ray diffraction (XRPD), polarized light microscopy (PLM), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and proton nuclear magnetic resonance (NPMR). 1 The starting material was characterized by H NMR. The results showed that it was crystalline and defined as Compound I·choline salt Form A, a monocholine salt (free acid:choline ratio 1:1).

[0275] Using Form A as the starting material, polymorph screening and single crystal growth were performed under 103 conditions by the methods of antisolvent addition, solid vapor diffusion, liquid vapor diffusion, evaporation, room temperature / 50°C slurrying, slow cooling, polymer-induced crystallization, and grinding. Based on the characterization results, only Form A of the choline salt of Compound I was obtained as a potential single crystal by liquid vapor diffusion.

[0276] Single crystal X-ray diffraction (SCXRD) was performed. Single crystal structure determination confirmed the chemical structure of Compound I·choline salt and showed that Form A is a nonstoichiometric hydrate of the monocholine salt.

[0277] Based on the results of the polymorph screening and single crystal studies described above, only Compound I·choline salt Form A was recommended for further pharmaceutical development.

[0278] Apparatus and method X-ray powder diffraction (XRPD) Powder X-ray diffraction data were collected under ambient conditions using a Bruker D2 PHASER diffractometer equipped with a 300 W low-power X-ray generator. Powder patterns were collected on a zero-background sample holder at 30 kV and 10 mA, with a total step of 1837 s, 2θ of 0.02° / step, and a Cu(Kα) X-ray tube, with a Kα2 / Kα1 intensity ratio of 0.50 (1.54439 Å / 1.5406 Å).

[0279] Thermogravimetric analysis (TGA) Thermogravimetric data were collected using a TA Discovery 550 series TGA. A few milligrams of material were heated from room temperature to the target temperature at a heating rate of 10°C per minute under nitrogen protection.

[0280] Differential scanning calorimetry (DSC) Differential scanning calorimetry was performed using a TA Discovery 2500 Series DSC on approximately a few milligrams of sample placed in a Tzero aluminum pan sealed with a Tzero airtight lid. Samples were analyzed under a nitrogen flow of 50 mL per minute using a heating rate of 10 °C per minute.

[0281] Polarized Light Microscope (PLM) Photomicrographs were taken at room temperature using an Olympus BX53M polarizing microscope.

[0282] Proton nuclear magnetic resonance ( 1 H NMR) 1 1 H NMR data was collected using an Agilent VNMR 400MR in D2O solvent.

[0283] Single Crystal X-ray Diffraction (SCXRD) Single crystal X-ray diffraction data were collected on a Rigaku mm007 Saturn70 (Mo / Kα X-ray, λ=0.71073 Å) diffractometer equipped with a Saturn70 detector, a fixed-chi goniometer, and a Rigaku GN2 cryosystem.

[0284] Solvent abbreviations The abbreviations for the solvents used are listed in Table 5-1. [Table 14]

[0285] Characterization of starting materials The starting material (Compound I·choline salt Form A) was yellow in color with a chemical purity of 99.9 area% and analyzed as received by XRPD, PLM, TGA, and DSC. XRPD data indicated Compound I·choline salt Form A. Birefringent crystals were observed by PLM. TGA showed a 1.3 wt% weight loss at 180.0 °C, and DSC showed a sharp melting peak at 199.7 °C (peak) with a broad endotherm at 104.0 °C (peak). 1 In H NMR (D2O), the stoichiometry (free acid:choline) was 1:1 and no solvent signal was observed.

[0286] Approximate solubility The solubility of the starting material (Compound I choline salt Form A) was estimated at room temperature (approximately 26 °C). Specifically, approximately 2 mg of solid was placed in an HPLC glass vial. Next, the solvents listed in Table 5-2 were added stepwise to the vial (50 / 50 / 200 / 700 μL) until the solid dissolved or the total volume reached 1.0 mL. The solubility data served as a guide for solvent selection in polymorph screening and single crystal growth. [Table 15]

[0287] Polymorph screening and single crystal growth Polymorph screening and single crystal growth were performed using the starting material (compound I·choline salt form A) under 103 conditions by antisolvent addition, solid vapor diffusion, liquid vapor diffusion, slow evaporation, room temperature / 50°C slurrying, slow cooling, polymer-induced crystallization, and grinding.

[0288] The crystallization methods and results are summarized in Table 5-3. Characterization results showed that single crystals of Compound I·choline salt Form A were obtained by liquid vapor diffusion under four conditions. [Table 16]

[0289] Addition of anti-solvent A total of 17 anti-solvent addition experiments were performed at room temperature. Stock solutions with the following corresponding solvents were prepared. A specific amount of starting material (Compound I·choline salt Form A) was dissolved in the solvent to obtain a solution / suspension, which was then filtered using a nylon membrane (pore size 0.22 μm). The filtrate was divided into approximately 50 mg of starting material for each experiment. The filtrate was magnetically stirred, and then 0.2–0.5 mL of anti-solvent was added stepwise until a precipitate appeared or the total volume of anti-solvent reached 15.0 mL. The resulting precipitate was isolated and analyzed by XRPD. The results in Table 5-4 indicate that only one crystalline form of Compound I·choline salt Form A was obtained. [Table 17]

[0290] solid vapor diffusion Solid vapor diffusion experiments were conducted using nine different solvents. Approximately 25 mg of starting material (compound I choline salt form A) was weighed into a 4 mL vial and placed in a 20 mL vial with approximately 3 mL of volatile solvent. The 20 mL vial was sealed with a cap and kept at room temperature for 8-10 days to allow the solvent vapor to interact with the sample. The solids were examined by XRPD, and the results are shown in Table 5-5. [Table 18]

[0291] Liquid Vapor Diffusion Liquid vapor diffusion experiments were performed under 10 solvent conditions. 20–50 mg of starting material (compound I·choline salt form A) was dissolved in 0.3–0.5 mL of the corresponding solvent. The solution / suspension was filtered using a nylon membrane (pore size 0.22 μm). The filtrate was then collected in a 4 mL vial covered with a pinhole cap and placed in a 20 mL vial with approximately 4 mL of volatile solvent. The 20 mL vial was sealed with a cap and kept at room temperature for 1–23 days to allow the solvent vapor to interact with the sample. The solids were examined by XRPD, and the results in Tables 5–6 show that only compound I·choline salt form A was obtained, four of which were potentially single crystals as indicated by PLM. [Table 19]

[0292] Slow evaporation 40-50 mg of starting material (Compound I choline salt form A) was dissolved in 2-5 mL of solvent in a 4 mL glass vial. The solution / suspension was filtered using a nylon membrane (pore size 0.22 μm). The filtrate was covered with a lid containing one pinhole and allowed to evaporate at room temperature. The results, summarized in Tables 5-7, showed that only a gel / oil was observed. [Table 20]

[0293] Slurry at room temperature Slurry experiments were performed at room temperature in 22 solvent / mixtures. Approximately 50 mg of starting material (Compound I·choline salt Form A) was suspended in 0.5 mL of solvent / mixture in an HPLC vial. The suspension was magnetically stirred at room temperature for 5 days, after which the remaining solid was isolated and analyzed by XRPD. The results are summarized in Tables 5-8. [Table 21]

[0294] Slurry at 50℃ Slurry experiments were performed at 50 °C in 18 solvents / mixtures. Approximately 50 mg of starting material (Compound I choline salt Form A) was suspended in 0.35–0.5 mL of solvent / mixture in an HPLC vial. The suspension was stirred at 50 °C for approximately 1–6 days, after which the remaining solid was isolated and subjected to XRPD analysis. The results are shown in Tables 5–9. [Table 22]

[0295] Slow cooling Slow cooling experiments were performed with eight solvent systems. Approximately 30–40 mg of starting material (Compound I·choline salt Form A) was equilibrated in 1.5–3.5 mL of solvent / mixture in a 4 mL glass vial at 50 °C for approximately 2 hours. The solution / suspension was filtered using a nylon membrane (pore size 0.22 μm), and the filtrate was slowly cooled to 5 °C at a rate of 0.1 °C / min. The results are summarized in Tables 5–10. [Table 23]

[0296] Polymer-induced crystallization Approximately 40-50 mg of starting material (compound I choline salt form A) was dissolved in 1.0-3.0 mL of solvent in a 4 mL glass vial. The solution / suspension was filtered using a nylon membrane (pore size 0.22 μm), and the filtrate was collected. 1-2 mg of a polymer mixture of PVC and PVP was added to the filtrate, and the sample was covered with a lid containing one pinhole and subsequently allowed to evaporate at room temperature. The results are summarized in Tables 5-11. [Table 24]

[0297] crushing Approximately 20 mg of starting material (Compound I choline salt Form A) was ground in a mortar with 20–40 μL of the corresponding solvent for 3–5 min. The solid was then characterized via XRPD. The results are summarized in Tables 5–12. [Table 25]

[0298] Single crystal structure determination A suitable single crystal of Compound I choline salt Form A of good diffraction quality was cut from the block crystal and selected for single-crystal X-ray diffraction. The crystal system of the single crystal was monoclinic and the space group was P21 / c. The unit cell parameters were {a = 8.4179(3) Å, b = 22.3688(7) Å, c = 14.7788(6) Å, α = 90°, β = 92.803(3)°, γ = 90°, V = 2779.49(17) Å}. 3 Further crystallographic data and refinement parameters are listed in Tables 5-13.

[0299] The asymmetric unit of the crystal structure of compound I choline salt form A consists of one free acid anion, one choline cation, and 0.21 water molecules, indicating that it is a monocholine salt. Static disorder of the choline moiety was observed within the asymmetric unit, resulting in the existence of two components (Part I and Part II, i.e., two conformations of the choline cation). The occupancies of Part I and Part II calculated by Olex2 were 0.79 and 0.21, respectively. It was confirmed that water molecules coexist when the choline cation was present in Part II, whereas no water molecules were observed in Part I. The occupancy of the water molecules was calculated to be 0.21, which is the same as the occupancy of the choline cation in Part II. In Part II, when the choline cation was oriented in conformation II, voids were found to occur at the positions of water molecules (calculated by Mercury with a probe radius of 1.0 Å and an approximate lattice spacing of 0.5 Å. Result: void volume of 29.31 ų. The volume of a water molecule in bulk water was 29.7 Å. Reference: Gerstein M, Chothia C., Proc Nati Acad Sci., 1996 (19), 10167-10172). In contrast, no voids were found in Part I, where the choline cation was oriented in conformation I. Because the proportion of conformation II influences the proportion of water molecules in the crystal lattice, the nature of compound I·choline salt form A was a nonstoichiometric hydrate. When the choline cation was oriented as in Part I, the free acid anion and the choline cation were connected to each other by an OH···N hydrogen bond. In Part II, an OH···O hydrogen bonding interaction was observed instead.

[0300] To determine whether the water content of Compound I choline salt Form A could be reduced to a lower level, an in situ TGA heating experiment was performed. Compound I choline salt Form A exhibited a gradual water loss starting at approximately room temperature, characteristic of a non-stoichiometric hydrate. After heating to 180 °C under N2, the sample was cooled and subjected to a second round of heating to remove volatiles. A low weight loss of 0.03 wt%, i.e., a water / Compound I molar ratio of 0.01, was observed. XRPD data for the starting material collected after heating to 180 °C showed no morphological change, in good agreement with a non-stoichiometric hydrate.

[0301] The atomic thermal ellipsoid plot (ORTEP diagram) of the crystal structure of compound I·choline salt form A is shown in Figure 12 (labeled as Part I) and confirms the proposed chemical structure (atomic thermal ellipsoids are drawn at a 50% probability level). Furthermore, no significant electron density peak was observed around the N6 atom, suggesting the transfer of an acidic proton and demonstrating that compound I is a choline salt. This is supported by the N-S and N-Ar bond lengths. Deprotonation of the R-SO2-NH-Ar moiety increases the resonance intensities of both the N-Ar and N-SO2 functional groups, shortening the bond lengths of the N-C and N-S bonds. The d(N-S) and d(N-C) bond lengths are 1.5811(12) Å and 1.3634(19) Å, respectively, also suggesting deprotonation of N6. [Table 26] [Table 27]

[0302] conclusion Based on the characterization results, compound I·choline salt form A was obtained and its potential single crystal was observed. Single crystal structure determination confirmed the chemical structure of compound I·choline salt and showed that form A is a nonstoichiometric hydrate (channel hydrate) of the monocholine salt.

[0303] Example 6: Pharmacokinetic study of Compound I·Choline Salt Form A and Free Acid Form A in dogs The objective of this study was to characterize the pharmacokinetics (PK) of Compound I in male beagle dogs after oral (PO) administration. PO formulations were prepared as capsules or dissolved in 0.5% or 1% methylcellulose (MC) aqueous solutions at a concentration of 6 mg / mL and administered orally at 30 mg / kg.

[0304] Apparatus and method LC-MS / MS for the PK study was performed using the following equipment / conditions. [Table 28]

[0305] Dosage Formulation Compound I free acid Form A and Compound I·choline salt Form A were formulated as oral suspensions at a target dose concentration of 6 mg / mL in 0.5% or 1% MC in water.

[0306] The target dose was 5 mL / kg and administered orally by gavage. Compound I choline salt Form A was also formulated as an oral capsule at a target dose concentration of 30 mg / kg. The formulation was prepared on the day of dosing and stored at room temperature prior to administration. Dogs were administered the drug by oral gavage. After oral administration, blood samples were collected by oral gavage at pre-dose and 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-dose.

[0307] Plasma concentrations of Compound I were measured by liquid chromatography with mass spectrometric detection (LC-MS / MS).

[0308] result Pharmacokinetic data are summarized in Table 6-3. [Table 29]

[0309] conclusion Compound I choline salt Form A demonstrated an excellent pharmacokinetic profile in both suspension and capsule formulations. Mean AUC of Compound I choline salt Form A in suspension or capsule formulations last Value and C max The values ​​were greater than those of Compound I free acid Form A administered as a suspension. Thus, Compound I choline salt Form A exhibited better oral bioavailability than Compound I free acid Form A.

[0310] Those skilled in the art will readily appreciate that the present disclosure is well adapted to obtain the ends and advantages mentioned, as well as those inherent therein. The methods, modifications, and compositions described herein, as currently representative of the subject matter, are exemplary and are not intended to limit the scope of the disclosure. Modifications therein and other uses which may occur to those skilled in the art are encompassed within the spirit of the disclosure and defined by the scope of the claims.

Claims

1. formula: 【Chemistry 1】 N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt (Compound I choline salt) having the formula:

2. A crystalline form of N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt (Compound I choline salt) or a solvate thereof.

3. Crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt Form A (Compound I choline salt Form A), characterized by X-ray powder diffraction with peaks at ±0.2 degrees 2θ selected from 12.9, 14.6, and 18.1 as measured on a diffractometer using Cu-Kα radiation.

4. 4. The crystalline Compound I choline salt Form A of claim 3, further characterized by an X-ray powder diffraction spectrum containing one or more additional peaks at ±0.2 degrees 2θ selected from 9.9, 16.7, 17.5, 19.8, 22.7, and 23.1 as measured on a diffractometer using Cu-Kα radiation.

5. 5. The crystalline Compound I choline salt Form A of claim 3 or 4, further characterized by an X-ray powder diffraction pattern substantially as shown in Figure 1A.

6. 6. The crystalline Compound I choline salt Form A of any one of claims 3 to 5, further characterized by a DSC comprising an endotherm at about 194-200 (peak).

7. 7. The crystalline Compound I choline salt Form A of any one of claims 3 to 6, further characterized by a DSC substantially as shown in Figure 1B.

8. Unit cell parameters: a=8.4179(3)Å, b=22.3688(7)Å, c=14.7788(6)Å, α=90°, β=92.803(3)°, γ=90°, V=2779.49(17)Å 3 Crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt Form A (Compound I choline salt Form A), having the formula:

9. Crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt Form B (Compound I choline salt Form B), characterized by X-ray powder diffraction with peaks at ±0.2 degrees 2θ selected from 11.0, 14.5, and 19.2 as measured on a diffractometer using Cu-Kα radiation.

10. X-ray powder diffraction containing one or more additional peaks at ±0.2 degrees 2θ selected from 18.4, 18.7, 19.8, 21.6, 22.1, and 24.4 as measured on a diffractometer using Cu-Kα radiation; X-ray powder diffraction substantially as shown in Figure 2A; DSC containing peaks at 74.4°C (peak) and 172.1°C (peak); or DSC substantially as shown in FIG. 2B; 10. The crystalline Compound I choline salt Form B of claim 9, further characterized by:

11. Crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide Form B (Compound I free acid Form B), characterized by X-ray powder diffraction with peaks at ±0.2 degrees 2θ selected from 9.5, 13.8, and 15.2 as measured on a diffractometer using Cu-Kα radiation.

12. X-ray powder diffraction including one or more additional peaks at ±0.2 degrees 2θ selected from 14.2, 17.1, 19.6, 26.0, 26.9, and 27.9 degrees 2θ ±0.2 degrees 2θ measured on a diffractometer using Cu-Kα radiation; X-ray powder diffraction substantially as shown in Figure 4A; DSC containing peaks at 108.8°C (peak) and 230.4°C (peak); or DSC substantially as shown in FIG. 4B; 12. The crystalline Compound I free acid form B of claim 11, further characterized by:

13. formula: 【Chemistry 2】 [In the formula, X is sodium and n is 1; X is potassium and n is 1; X is calcium and n is 2; X is magnesium and n is 2. or a solvate thereof.

14. 14. The crystalline salt form of claim 13, wherein the crystalline salt form is selected from the group consisting of Compound I, sodium salt Form A, Compound I, potassium salt Form A, Compound I, potassium salt Form B, Compound I, calcium salt Form A, and Compound I, magnesium salt Form A.

15. 15. A pharmaceutical composition comprising Compound I • choline salt or a solvate thereof according to claim 1, a crystalline form of Compound I • choline salt according to any one of claims 2 to 10, crystalline Compound I according to claim 11 or 12, or a crystalline salt form according to claim 13 or 14, and a pharmaceutically acceptable excipient.

16. 16. The pharmaceutical composition of claim 15, wherein at least 99% of Compound I in the pharmaceutical composition is Compound I choline salt or a solvate thereof as defined in claim 1.

17. 16. The pharmaceutical composition of claim 15, wherein at least 99% of Compound I in the pharmaceutical composition is Compound I•choline salt or a solvate thereof as defined in claim 1, a crystalline form of Compound I•choline salt or a solvate thereof as defined in claim 2, or a crystalline form of Compound I•choline salt Form A as defined in any one of claims 3 to 8.

18. 16. The pharmaceutical composition of claim 15, wherein at least 99% of Compound I in the pharmaceutical composition is the crystalline form of Compound I choline salt Form B as defined in claim 9 or 10.

19. 16. The pharmaceutical composition of claim 15, wherein at least 99% of Compound I in the pharmaceutical composition is crystalline Compound I of claim 11 or 12.

20. 16. The pharmaceutical composition of claim 15, wherein at least 99% of Compound I in the pharmaceutical composition is a crystalline salt form according to claim 13 or 14.

21. 21. A method for treating a disease, disorder or condition in which inhibition or impairment of APJ receptor signaling or downregulation of endogenous apelin contributes to the pathology and / or symptoms and / or progression of the disease, disorder or condition, comprising administering to a subject in need thereof an effective amount of the pharmaceutical composition of any one of claims 15 to 20.

22. 22. The method of claim 21, wherein the disease, disorder, or condition is pulmonary arterial hypertension (PAH).

23. 23. The method of claim 22, wherein the PAH is idiopathic.

24. 23. The method of claim 22, wherein the PAH is hereditary PAH, toxin- or drug-induced PAH; or PAH associated with one or more of congenital heart disease, connective tissue disorders (e.g., scleroderma, systemic lupus erythematosus, systemic sclerosis, Hashimoto's thyroiditis, Sjogren's syndrome, and antiphospholipid syndrome), portal hypertension, BMPR2 mutations, schistosomiasis, or HIV infection.

25. 22. The method of claim 21, wherein the disease, disorder or condition is fibrosis.

26. 26. The method of claim 25, wherein the fibrosis is associated with an organ or tissue selected from the group consisting of lung, liver, heart, mediastinum, bone marrow, retroperitoneum, skin, intestine, joints and reproductive organs, or a combination thereof.

27. 26. The method of claim 21 or 25, wherein the disease, disorder or condition is idiopathic pulmonary fibrosis (IPF).

28. 22. The method of claim 21, wherein the disease, disorder or condition is a connective tissue disorder.

29. 29. The method of claim 28, wherein the connective tissue disorder is selected from the group consisting of scleroderma, systemic lupus erythematosus, systemic sclerosis, Hashimoto's thyroiditis, Sjogren's syndrome, and antiphospholipid syndrome.

30. 30. The method of claim 21, 28 or 29, wherein the disease, disorder or condition is systemic sclerosis.

31. 1. A method for preparing crystalline N-(1-(2,6-dimethoxyphenyl)-2-(6-ethoxypyridin-2-yl)-1H-imidazo[4,5-b]pyrazin-6-yl)methanesulfonamide choline salt (Compound I choline salt), comprising contacting Compound I or a salt thereof with choline in a solvent for a time sufficient to provide crystalline Compound I choline salt.

32. 32. The method of claim 31 , wherein the solvent is acetone.

33. 32. The method of claim 31, wherein the contacting comprises adding an equimolar amount of choline to Compound I at a temperature of about 0°C to about 50°C.

34. 32. The method of claim 31, further comprising isolating the crystalline Compound I choline salt after said contacting step.

35. 35. The method of claim 34, wherein isolating comprises filtering, washing, and drying the crystalline Compound I choline salt.

36. 36. The method of any one of claims 31-35, wherein at least about 95% of the crystalline Compound I choline salt is Form A.

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