Crystalline and salt forms of NLRP3 inhibitor

Abstract

The present disclosure provides crystalline forms of an NLRP3 inhibitor, as well as methods of making and using these forms.

Description

[0001] Related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 582,161, filed September 12, 2023, the entire contents of which are hereby incorporated. Background Technology

[0003] Aging frailty constitutes a very worrying problem for an individual’s overall health and well-being and is characterized as a syndrome of abnormal physiological regulation across multiple systems. Aging frailty is a senile syndrome characterized by frailty, reduced physical activity, decreased motor function, fatigue, and unconscious weight loss (Yao, X. et al., Clinics in Geriatric Medicine 27(1): 79-87 (2011)). Furthermore, many studies have shown a direct correlation between aging frailty and inflammation (Hubbard, RE et al., Biogerontology 11(5):635-641 (2010)). Immunosenescence is characterized by a low-grade, chronic systemic inflammatory state known as “inflammaging” (Franceshi, C. et al., Annals of the New York Academy of Sciences 908:244-254 (2000)). This enhanced inflammatory state or chronic inflammation found in aging and aging frailty leads to abnormal immune regulation and a complex remodeling of both innate and adaptive immunity.

[0004] Inhibition of the NLRP3 inflammasome (an oligomeric protein complex comprising ASC and caspase-1) mediates inflammation in numerous preclinical models (Schwaid, AG, J. Med. Chem. 2021, 64(1), 101-122). Meanwhile, the NLRP3 inflammasome is part of a larger pro-inflammatory pathway whose regulation is being explored. NLRP3 is an inflammasome sensor protein that has been well-studied in a variety of disease contexts. Many different indications are associated with the NLRP3 inflammasome, including age-related diseases, cold pyridine-associated periodic syndrome (CAPS), non-alcoholic steatohepatitis (NASH), gout, coronary artery disease, Crohn's disease, osteoarthritis, rheumatoid arthritis, Alzheimer's disease, Parkinson's disease, intestinal disorders, acute respiratory distress syndrome (ARDS), amyotrophic lateral sclerosis (ALS), cancer, and skin diseases.

[0005] Inflammation and activation of the NLRP3 inflammasome have also been shown to lead to hearing loss (Nakanishi, H. et al.). Frontiers in Neurology , 2020, 11 , 1-7; Nakanishi, H. et al., PNAS (2017, E7766-E7775). Inflammation-related hearing loss can be age-dependent (Fischer, N. et al., 2017, E7766-E7775). Gerontology (2019, 1-7), noise-induced (Le Prell, CG et al.) Current Opinion in Physiology , 2020, 18 (32-36) and the consequences of viral infections such as Zika virus and coronavirus (Yee, KT et al., Hearing Research , 2020, 395 , 1-15).

[0006] Therefore, the NLRP3 inflammasome is a promising drug target. The breadth of indications it is involved in indicates a need for therapeutics targeting the NLRP3 inflammasome. Summary of the Invention

[0007] This article provides the crystalline forms of compounds that inhibit the NLRP3 inflammasome. Therefore, these compounds can be used to treat a variety of indications, including inflammatory aging and inflammation.

[0008] In one particular aspect, this article provides the crystalline form of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide, referred to herein as compound 1: .

[0009] In another aspect, this article provides a method for treating diseases and conditions associated with the NLRP3 inflammasome in subjects of need, the method comprising administering to the subject a crystalline form of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide. Attached Figure Description

[0010] Figure 1 The XRPD diffraction pattern of crystalline compound 1 (pattern B) is shown.

[0011] Figure 2The DSC thermogram of crystalline compound 1 (type B) is shown.

[0012] Figure 3 The XRPD diffraction pattern of crystalline compound 1 hydrate (type A) is shown.

[0013] Figure 4 The DSC thermogram of crystalline compound 1-hydrate (type A) is shown.

[0014] Figure 5 The TGA thermogram of crystalline compound 1 hydrate (type A) is shown.

[0015] Figure 6 The XRPD diffraction pattern of crystalline compound 1 HCl salt (type A) is shown.

[0016] Figure 7 The DSC thermogram of crystalline compound 1 HCl salt (type A) is shown.

[0017] Figure 8 The XRPD diffraction pattern of crystalline compound 1 phosphate (type A) is shown.

[0018] Figure 9 The DSC thermogram of crystalline compound 1 phosphate (type A) is shown.

[0019] Figure 10 The TGA thermogram of crystalline compound 1 phosphate (type A) is shown.

[0020] Figure 11 The XRPD diffraction pattern of sodium salt of crystalline compound 1 (type A) is shown.

[0021] Figure 12 The DSC thermogram of sodium salt of crystalline compound 1 (type A) is shown.

[0022] Figure 13 The TGA thermogram of sodium salt of crystalline compound 1 (type A) is shown. Detailed Implementation

[0023] When a compound is used for pharmaceutical purposes, its solid state is important. Changing a compound from one solid form to another alters its physical properties, which can affect the suitability of that form for pharmaceutical use. For example, certain crystalline solid compounds can overcome the disadvantages of other solid forms, such as, for instance, instability and / or reduced purity.

[0024] This article provides the solid crystalline form of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide (compound 1): ; Or its pharmaceutically acceptable salts or hydrates.

[0025] This compound is disclosed in international application number PCT / US2022 / 021461, the entire contents of which are incorporated herein by reference.

[0026] The crystalline forms presented in this paper can be characterized by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA).

[0027] definition

[0028] The following lists definitions of various terms used to describe the crystalline forms provided herein. These definitions apply to the terms used individually or as part of the larger group throughout this specification and claims, unless otherwise limited in specific instances.

[0029] Unless otherwise defined, all technical and scientific terms used herein have, in general, the same meanings as commonly understood by one of ordinary skill in the art to which the compound and its crystalline forms pertain. In general, the nomenclature used herein, as well as laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry, are those well-known and commonly used in the art.

[0030] As used herein, the article “a / an” refers to one or more grammatical objects of the article (i.e., at least one). For example, “a / an element” means one or more elements. Furthermore, the use of the term “including” and other forms such as “include,” “includes,” and “included” is not restrictive.

[0031] As used in this article, the term "EC" 50 "This refers to the concentration of the compound required to achieve 50% of its maximum observed effect."

[0032] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, which participates in carrying or transporting, within or to a patient, a compound that may be used in this disclosure to perform its intended function. Typically, such constructs carry or transport from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense that it is compatible with other components of the formulation, including the compounds provided herein, and is harmless to the patient.

[0033] Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth gum; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; phosphate buffer solutions; and other non-toxic compatible substances used in pharmaceutical preparations.

[0034] As used herein, "pharmaceutically acceptable carrier" also includes any and all coatings, antibacterial and antifungal agents, and absorption delay agents that are compatible with the activity of the compounds provided herein and are physiologically acceptable to a patient. Additional active compounds may also be incorporated into the composition. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of this disclosure are known in the art and have been described, for example, in Remington's Pharmaceutical Sciences (edited by Genaro, Mack Publishing Co., 1985, Easton, PA), which are incorporated herein by reference.

[0035] As used herein, the phrases “therapeutic effective dose” and “therapeutic effective amount” refer to the amount of a compound that prevents the onset of disease, alleviates the symptoms of disease, halts the progression of disease, or produces another desired biological outcome, such as, for example, improvement of clinical signs.

[0036] The terms “treat,” “treated,” “treating,” or “treatment” include the reduction or relief of at least one symptom related to or caused by the condition, symptom, or disease being treated.

[0037] As used herein, the terms “prevent” or “prevention” mean the absence of symptoms or disease in the absence of symptoms or disease, or the absence of further symptoms or disease development in the presence of symptoms or disease. The ability to prevent some or all of the symptoms associated with the symptoms or disease must also be considered.

[0038] As used herein, the terms “patient,” “individual,” or “subject” refer to a human or a non-human mammal. Non-human mammals include, for example, livestock and pets such as sheep, cattle, pigs, dogs, cats, and rodents. In one implementation, the patient, subject, or individual is a human.

[0039] The terms "administering" or "administration," etc., refer to the provision of a therapeutic agent, such as the crystalline form disclosed herein, to a subject in need of treatment. In one embodiment, the subject is a mammal. In another embodiment, the subject is a human.

[0040] As used herein, the term “about” will be understood by those skilled in the art and will vary to some extent depending on the context in which the term is used. As used herein, when referring to measurable values ​​such as quantity, duration, etc., the term “about” is intended to cover a variation of ±10% (inclusive of ±5%, ±1%, and ±0.1%) relative to a specified value, and such variation is suitable for performing the disclosed methods.

[0041] Characterization of crystalline form

[0042] In some embodiments, the crystalline form described herein can be identified based on characteristic peaks in X-ray powder diffraction analysis. X-ray powder diffraction (XRPD) is a scientific technique for characterizing the structure of powders, crystallites, or other solid materials using X-rays, neutrons, or electron diffraction. Descriptions of methods for obtaining certain XRPD diffraction patterns associated with the crystalline forms provided herein can be found in the following examples. In one embodiment, the X-ray powder diffraction data provided herein are obtained using Cu Kα radiation.

[0043] In one respect, this article provides crystalline form of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide or its pharmaceutically acceptable salts or hydrates.

[0044] In one embodiment, the crystalline form is a monohydrate. In another embodiment, the crystalline form is anhydrous.

[0045] In yet another embodiment, the crystalline form is a pharmaceutically acceptable salt. In still another embodiment, the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, phosphate, and sodium salt.

[0046] Anhydrous free form

[0047] In one embodiment, compound 1 is in anhydrous crystalline form.

[0048] In one embodiment, the crystalline form is characterized by XRPD diffraction patterns having peaks at angles (±0.2 degrees) of 7.2, 20.4, and 20.6 degrees, expressed in 2-θ degrees. In another embodiment, the crystalline form is characterized by XRPD diffraction patterns having peaks at angles (±0.2 degrees) of 7.2, 20.4, 20.6, and 21.6 degrees, expressed in 2-θ degrees. In yet another embodiment, the crystalline form is characterized by XRPD diffraction patterns having peaks at angles (±0.2 degrees) of 7.2, 11.1, 11.8, 20.4, 20.6, and 21.6 degrees, expressed in 2-θ degrees. In yet another embodiment, the crystalline form is characterized by XRPD diffraction patterns having peaks at angles (±0.2 degrees) of 5.5, 7.2, 11.1, 11.8, 15.3, 20.4, 20.6, 21.6, 26.7, and 30.9 degrees, expressed in 2-θ degrees.

[0049] In one embodiment, the crystalline form is characterized by an XRPD diffraction pattern showing peaks selected from Table 1 (Type B).

[0050] Table 1

[0051] XRPD peak table of free form type B

[0052] In another embodiment, the crystalline form has essentially the following characteristics: Figure 1 The depicted XRPD diffraction pattern.

[0053] In another embodiment, the crystalline form is a monohydrate, which has a DSC thermogram characterized by an endothermic onset temperature of 177.3°C. In yet another embodiment, the crystalline form has substantially the following characteristics: Figure 2 The depicted DSC thermogram.

[0054] Free form monohydrate

[0055] In one embodiment, the crystalline form is a monohydrate, characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 6.5, 12.9, and 16.1 degrees, expressed in 2-θ degrees. In another embodiment, the crystalline form is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 6.5, 11.2, 12.9, and 16.1 degrees, expressed in 2-θ degrees. In yet another embodiment, the crystalline form is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 6.5, 11.2, 12.9, 16.1, 22.7, 24.8, and 32.6 degrees, expressed in 2-θ degrees.

[0056] In one embodiment, the crystalline form is characterized by an XRPD diffraction pattern showing peaks selected from Table 2 (Type A).

[0057] Table 2

[0058] XRPD peak table of free form type A

[0059] In another embodiment, the crystalline form has essentially the following characteristics: Figure 3 The depicted XRPD diffraction pattern.

[0060] In another embodiment, the crystalline form is a monohydrate, said monohydrate having a DSC thermogram characterized by an endothermic onset temperature of 139.6°C. In yet another embodiment, the crystalline form has substantially the following characteristics: Figure 4 The depicted DSC thermogram. In yet another embodiment, the crystalline form has essentially the same... Figure 5 The depicted TGA thermogram.

[0061] hydrochloride

[0062] In one embodiment, the crystalline form is 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide hydrochloride. In another embodiment, the crystalline form of the HCl salt is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 12.3, 18.1, and 18.2 degrees, expressed in 2-θ degrees. In yet another embodiment, the crystalline form of the HCl salt is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 12.3, 12.8, 18.1, 18.2, and 28.7 degrees, expressed in 2-θ degrees. In another embodiment, the crystalline form of the HCl salt is characterized by XRPD diffraction patterns having peaks at angles (±0.2 degrees) of 12.3, 12.8, 18.1, 18.2, 26.0, and 28.7 degrees, expressed in 2-θ degrees.

[0063] In one embodiment, the crystalline form is characterized by an XRPD diffraction pattern showing peaks selected from Table 3 (HCl salt of type A).

[0064] Table 3

[0065] XRPD peak table of HCl salt form A

[0066] In another embodiment, the crystalline form has essentially the following characteristics: Figure 6 The depicted XRPD diffraction pattern.

[0067] In another embodiment, the crystalline form is an HCl salt, said HCl salt having a DSC thermogram characterized by an endothermic onset temperature of 218.3°C. In yet another embodiment, the crystalline form has substantially the following characteristics: Figure 7 The depicted DSC thermogram.

[0068] phosphate

[0069] In one embodiment, the crystalline form is 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide phosphate.

[0070] In another embodiment, the crystalline form of the phosphate is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 3.1, 9.2, and 16.4 degrees, expressed in 2-θ degrees. In yet another embodiment, the crystalline form of the phosphate is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 3.1, 9.2, 11.7, 13.9, 15.4, 16.4, and 24.7 degrees, expressed in 2-θ degrees. In yet another embodiment, the crystalline form of the phosphate is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 3.1, 9.2, 11.7, 13.9, 15.4, 16.4, 18.1, 19.0, 16.4, and 24.7 degrees, expressed in 2-θ degrees.

[0071] In one embodiment, the crystalline form is characterized by an XRPD diffraction pattern showing peaks selected from Table 4 (phosphate of type A).

[0072] Table 4

[0073] XRPD peak table of phosphate form A

[0074] In another embodiment, the crystalline form has essentially the following characteristics: Figure 8 The depicted XRPD diffraction pattern.

[0075] In another embodiment, the crystalline form is a phosphate, which has a DSC thermogram characterized by an endothermic onset temperature of 223.9°C. In another embodiment, the crystalline form has essentially the following characteristics: Figure 9 The depicted DSC thermogram. In yet another embodiment, the crystalline form has essentially the same... Figure 10 The depicted TGA thermogram.

[0076] Sodium salt

[0077] In one embodiment, the crystalline form is sodium 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide.

[0078] In another embodiment, the crystalline form of the sodium salt is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 10.3, 21.1, and 25.8 degrees, expressed in 2-θ degrees. In yet another embodiment, the crystalline form of the sodium salt is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 10.3, 17.3, 20.6, 21.1, 23.3, and 25.8 degrees, expressed in 2-θ degrees. In still another embodiment, the crystalline form of the sodium salt is characterized by XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 10.3, 14.9, 17.2, 17.3, 18.8, 20.6, 20.0, 21.1, 23.3, and 25.8 degrees, expressed in 2-θ degrees.

[0079] In one embodiment, the crystalline form is characterized by an XRPD diffraction pattern showing peaks selected from Table 5 (sodium salt of type A).

[0080] Table 5

[0081] XRPD peak table of sodium salt type A

[0082] In another embodiment, the crystalline form has essentially the following characteristics: Figure 11 The depicted XRPD diffraction pattern.

[0083] In another embodiment, the crystalline form has essentially the following characteristics: Figure 12 The depicted DSC thermogram. In yet another embodiment, the crystalline form has essentially the same... Figure 13 The depicted TGA thermogram.

[0084] Pharmaceutically acceptable salts

[0085] In another respect, this article provides crystalline forms of pharmaceutically acceptable salts of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide, wherein the pharmaceutically acceptable salts are selected from the group consisting of hydrochloride, phosphate and sodium salts.

[0086] In one embodiment, the pharmaceutically acceptable salt is a hydrochloride salt. In another embodiment, the pharmaceutically acceptable salt is a phosphate salt. In yet another embodiment, the pharmaceutically acceptable salt is a sodium salt.

[0087] In one embodiment, the X-ray powder diffraction (XRPD) of the crystalline form herein is measured by Cu Kα radiation operated at 40 kV, 40 mA. In another embodiment of XRPD, the scan is run from 2–40 degrees 2–θ in steps of 0.02 degrees and a scan time of 0.3 seconds per step.

[0088] In another aspect, this document provides a pharmaceutical composition comprising the crystalline form of the present disclosure and a pharmaceutically acceptable carrier.

[0089] The pharmaceutical preparations disclosed herein can be prepared according to standard procedures and administered at doses selected to alleviate, prevent, or eliminate disease. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, and Goodman and Gilman's “The Pharmaceutical Basis of Therapeutics,” Pergamon Press, New York, NY (the contents of which are incorporated herein by reference), for a general description of methods of administering various agents for human treatment.

[0090] The pharmaceutical compositions described herein may comprise the crystalline form disclosed herein combined with one or more non-toxic, pharmaceutically acceptable carriers and / or diluents and / or adjuvants and / or excipients.

[0091] For oral or parenteral administration, the crystalline forms disclosed herein can be mixed with conventional drug carriers and excipients and used in the form of tablets, capsules, elixirs, suspensions, syrups, rice paper capsules, etc. Compositions comprising the crystalline forms disclosed herein may contain from about 0.1% to about 99% by weight of the active compound, such as from about 10% to about 30%.

[0092] For oral administration, solid dosage forms such as tablets and capsules are available. Sustained-release or enteric-coated formulations can also be designed. For pediatric and geriatric applications, one embodiment provides suspensions, syrups, and chewable tablets. For oral administration, the pharmaceutical composition is in, for example, the form of tablets, capsules, suspensions, or liquids.

[0093] Pharmaceutical compositions can be formulated into dosage units containing a therapeutically effective amount of the active ingredient. Examples of such dosage units are tablets and capsules. For therapeutic purposes, tablets and capsules may contain, in addition to the active ingredient, conventional carriers such as binders, fillers, lubricants, disintegrants, or acceptable wetting agents. Oral liquid formulations are generally in the form of aqueous or oily solutions, suspensions, emulsions, syrups, or elixirs.

[0094] The pharmaceutical compositions disclosed herein can be placed in a pharmaceutically acceptable carrier and delivered to a receiving subject (e.g., a human) according to known methods of drug delivery. Generally, methods of delivering pharmaceutical compositions in vivo utilize protocols for delivery agents recognized in the art, wherein the only substantial procedural modification is the substitution of the drug in the crystalline form of the present disclosure for the drug in the protocols recognized in the art.

[0095] Treatment

[0096] This article provides a method for treating a disease, the method comprising administering crystalline form of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide, or a pharmaceutical composition comprising said crystalline form and a pharmaceutically acceptable carrier.

[0097] In one aspect, a method for inhibiting NLRP3 inflammasomes in a subject in need is provided, the method comprising administering to the subject a therapeutically effective amount of the crystalline form disclosed herein.

[0098] In another aspect, a method for treating inflammation in a subject in need is provided, the method comprising administering to the subject a therapeutically effective amount of the crystalline form disclosed herein.

[0099] In another aspect, a method for treating inflammatory aging in a subject of need is provided, the method comprising administering to the subject a therapeutically effective amount of the crystalline form disclosed herein.

[0100] In another aspect, a method is provided for treating a subject with cold pyridine-associated periodic syndrome (CAPS), the method comprising administering to the subject a therapeutically effective amount of the crystalline form disclosed herein.

[0101] In one implementation, CAPS is selected from the group consisting of: familial cold autoinflammatory syndrome, Muckle-Wells syndrome, and neonatal episodic multisystem inflammatory disease.

[0102] In one aspect, a method is provided for treating an inner ear disease or condition in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the crystalline form disclosed herein.

[0103] In one implementation, inner ear diseases or conditions are selected from the group consisting of: hearing loss, hearing impairment, vertigo, Meniere's disease, and tinnitus.

[0104] In another aspect, this article provides a method for treating a skin condition in a subject in need, the method comprising administering to an individual a therapeutically effective amount of the crystalline form disclosed herein.

[0105] In one implementation, the skin disease is selected from the group consisting of: psoriasis, urticaria, photoaging of the skin, and eczema.

[0106] This article also provides a method for treating or alleviating aging or aging-related disorders that adversely affect lifespan or quality of life using the crystalline form provided herein, wherein the aging-related disorders adversely affecting lifespan or quality of life are selected from the group consisting of: inflammation, anemia, hyperglycemia, dyslipidemia, hyperinsulinemia, insulin resistance, immunosuppression, liver disease, iron overload, hypertriglyceridemia, impaired skin integrity, wound healing, scarring, pain, allergies, sleep disorders and problems, gastrointestinal disorders and problems, Th1 inflammation, Th2 inflammation, inflammatory diseases involving T cell-dependent B cell proliferation, T cell-dependent B cell proliferation, allergies, asthma, atherosclerosis, autoimmune diseases, hypercholesterolemia, chronic inflammation, chronic obstructive pulmonary disease (COPD), Crohn's disease, skin reactions to tissue damage, fibrosis, hematologic oncology, metabolic diseases, and cardiovascular diseases. Organ transplantation, psoriasis, liver fibrosis, dermatitis, pulmonary fibrosis, pulmonary response to respiratory infections, restenosis, rheumatoid arthritis, sarcomatoid diseases, matrix biology of tumors, systemic lupus erythematosus (SLE), ulcerative colitis, vasculitis, and diseases driven or exacerbated by one or more factors selected from the group consisting of: α-smooth muscle protein (αSMA), CD40, CD69, collagen I, collagen III, decorin, e-selectin, and eosinophil chemokine 3. (CCL26), fibroblast proliferation, human leukocyte antigen-DR isotype (HLA-DR), immunoglobulin G, interferon-γ induced protein 10 (IP-10 / CXCL10), interferon-induced T cell α chemokine (I-TAC / CXCL11), interleukin (IL)-1, IL-1α, IL-2, IL-6, IL-8 (CXCL8), IL-10, IL-17A, IL-17F, keratin 8 / 81, macrophage community-stimulating factor (M-CSF), matrix metalloproteinase (MMP)-1, MMP-9, monocyte chemokine protein 1 (MCP-1), interferon-γ induced mononuclear factor (MIG / CXCL9), plasminogen activator inhibitor 1 (PAI-1), prostaglandin E2 (PGE2), serum amyloid A, T or B cell proliferation, tissue plasminogen activator (tPA), tumor necrosis factor α (TNF.α.), vascular cell adhesion molecule (VCAM-1) and vascular endothelial growth factor 2 (VEGFR2), the method comprising: administering the crystalline form provided herein to a subject in need.

[0107] In one aspect, this article provides a method for reversing the natural aging process of a subject, the method comprising administering to the subject a therapeutically effective amount of the crystalline form provided herein or a pharmaceutically acceptable salt thereof.

[0108] In another aspect, this article provides a method for reversing the natural aging process of a subject, the method comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of the crystalline form or a pharmaceutically acceptable salt thereof provided herein.

[0109] In another aspect, this article provides a method for prolonging the life of a subject, the method comprising administering to the subject a therapeutically effective amount of the crystalline form provided herein or a pharmaceutically acceptable salt thereof.

[0110] In another aspect, this article provides a method for prolonging the life of a subject, the method comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form or a pharmaceutically acceptable salt thereof provided herein.

[0111] In another aspect, this article provides a method for slowing and mitigating the aging process in a subject, the method comprising administering to the subject a therapeutically effective amount of the crystalline form provided herein or a pharmaceutically acceptable salt thereof.

[0112] In another aspect, this document provides a method for inhibiting or modulating pro-inflammatory pathways in cells, the method comprising contacting the cells with a crystalline form provided herein or a pharmaceutically acceptable salt thereof. In yet another aspect, this document provides a method for inhibiting or modulating NLRP3 in cells, the method comprising contacting the cells with a crystalline form provided herein or a pharmaceutically acceptable salt thereof.

[0113] Treating cells expressing the NLRP3 inflammasome (in vitro or in vivo) with the crystalline form provided in this paper can suppress pro-inflammatory pathways and inhibit downstream events associated with signaling pathways, such as inflammation or inflammatory senescence.

[0114] In another aspect, this article provides a method for treating sensory nerve disorders in subjects in need, the method comprising administering to an individual a therapeutically effective amount of the crystalline form disclosed herein.

[0115] In one implementation, sensory nerve diseases are selected from the group consisting of: hearing loss, hearing impairment, and eye diseases. In one implementation, eye diseases are retinal and optic nerve damage.

[0116] In another aspect, this article provides a method for treating an inflammatory condition in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the crystalline form disclosed herein.

[0117] In one implementation scheme, inflammatory conditions are selected from the group consisting of: allergies, asthma, atopic dermatitis, atherosclerosis, autoimmune diseases, celiac disease, chronic inflammation, glomerulonephritis, hepatitis, inflammatory bowel disease, reperfusion injury, SARS-CoV-2 infection, transplant rejection, heart disease, diabetes, arthritis, Crohn's disease, ulcerative colitis, non-alcoholic steatosis (NASH), gout, coronary artery disease, rheumatoid arthritis, intestinal diseases, and acute respiratory distress syndrome (ARDS).

[0118] In another embodiment, the inflammatory condition is a neuroinflammatory disease. In another embodiment, the inflammatory condition is inner ear inflammation.

[0119] In one implementation, chronic inflammation includes tissue inflammation. Tissue inflammation is chronic inflammation limited to a specific tissue or organ. In one implementation, tissue inflammation includes, for example, inflammation of the skin, eyes, muscles, tendons, ligaments, bones, cartilage, lungs, heart, liver, pancreas, kidneys, bladder, stomach, intestines, neurons, and brain.

[0120] In another implementation, chronic inflammation includes systemic inflammation. While the processes involved are the same as in tissue inflammation, systemic inflammation is not limited to a specific tissue but rather damages the entire body, involving the endothelium and other organ systems. The term sepsis is appropriate when it is attributed to infection; bacteremia is appropriate for bacterial sepsis, and viremia is appropriate for viral sepsis. Vasodilatory dysfunction and organ dysfunction are serious problems associated with widespread infection and can lead to septic shock and death.

[0121] In yet another implementation, chronic inflammation includes arthritis. Arthritis encompasses a group of conditions involving damage to joints in the body due to inflammation of the synovium, including but not limited to osteoarthritis, rheumatoid arthritis, juvenile idiopathic arthritis, spondyloarthritis-like ankylosing spondylitis, reactive arthritis (Reiter's syndrome), psoriatic arthritis, enteropathic arthritis associated with inflammatory bowel disease, Whipple's disease, and Behcet's disease, septic arthritis, gout (also known as gouty arthritis, crystalline synovitis, or metabolic arthritis), pseudogout (calcium pyrophosphate deposition disease), and Still's disease. Arthritis can affect a single joint (monoarthritis), two to four joints (oligoarthritis), or five or more joints (polyarthritis) and can be an autoimmune or non-autoimmune disease.

[0122] In another aspect, this article provides a method for treating age-related conditions in subjects in need, the method comprising administering to an individual a therapeutically effective amount of the crystalline form disclosed herein.

[0123] In one implementation plan, age-related conditions are selected from the group consisting of: neurodegeneration, cardiovascular disease, insulin resistance, diabetes, osteoporosis, osteoarthritis, cognitive decline, dementia, fragility, cataracts, arthritis, obesity, hypertension, angina pectoris, congestive heart failure, dyslipidemia, myocardial infarction, vascular disease, respiratory disease, kidney disease, cerebrovascular disease, peripheral vascular disease, Alzheimer's disease, diastolic dysfunction, benign prostatic hyperplasia, aortic aneurysm, and emphysema.

[0124] In another aspect, this article provides a method for treating metabolic disorders in subjects in need, the method comprising administering to an individual a therapeutically effective amount of the crystalline form disclosed herein.

[0125] In one implementation scheme, metabolic disorders are selected from the group consisting of: diabetes, obesity, cystic fibrosis, and hyperthyroidism.

[0126] In another aspect, this article provides a method for treating neurodegenerative diseases in subjects of need, the method comprising administering to an individual a therapeutically effective amount of the crystalline form disclosed herein.

[0127] In one implementation scheme, neurodegenerative diseases are selected from the group consisting of: Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), and Batten's disease.

[0128] In one aspect, this article provides a method for treating an inner ear disease or condition in a subject in need, the method comprising administering to an individual a therapeutically effective amount of the crystalline form disclosed herein.

[0129] In one implementation, the inner ear disease or condition is selected from the group consisting of: hearing loss, hearing impairment, vertigo, Meniere's disease, and tinnitus. In another implementation, the inner ear disease is hearing loss. In yet another implementation, the inner ear disease is hearing impairment.

[0130] In another implementation, the hearing loss is age-related, noise-induced, or a result of a viral infection. In yet another implementation, the viral infection is Zika virus or coronavirus.

[0131] In another aspect, this disclosure provides a crystalline form disclosed herein or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for treating or preventing diseases in which the NLRP3 inflammasome plays a role.

[0132] In one aspect, this article provides a method for treating diseases selected from the group consisting of: autoimmune diseases, inflammatory diseases, proliferative and hyperproliferative diseases, immune-mediated diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cardiovascular diseases, hormone-related diseases, allergies, asthma, and Alzheimer's disease. In other embodiments, the diseases are selected from proliferative disorders and neurodegenerative disorders.

[0133] One aspect of this disclosure provides crystalline forms of compounds that can be used to treat diseases, conditions, and disorders characterized by excessive or abnormal cell proliferation. Such diseases include, but are not limited to, proliferative or hyperproliferative disorders and neurodegenerative diseases. Examples of proliferative and hyperproliferative disorders include, but are not limited to, cancer.

[0134] Therefore, in one aspect, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the crystalline form disclosed herein or a pharmaceutically acceptable salt thereof.

[0135] In one implementation plan, cancers are selected from the group consisting of: breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, urogenital tract cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, skin cancer, keratoacanthoma, lung cancer, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, colorectal cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder cancer, hepatocellular carcinoma and biliary tract cancer, kidney cancer, myeloid diseases, lymphoid diseases, Hodgkin's disease, pilonidal carcinoma, buccal and pharyngeal (oral) cancer, lip cancer, tongue cancer, oral cancer, pharyngeal cancer, small bowel cancer, colon cancer, rectal cancer, brain cancer and central nervous system cancer, chronic myeloid leukemia (CML) and leukemia.

[0136] In another implementation, the cancer is selected from the group consisting of myeloma, lymphoma, or selected from the group consisting of: gastric cancer, kidney cancer, head and neck cancer, oropharyngeal cancer, non-small cell lung cancer (NSCLC), endometrial cancer, liver cancer, non-Hodgkin's lymphoma, and lung cancer.

[0137] In one implementation scheme, the cancer is selected from the group consisting of: prostate cancer, colon cancer, lung cancer, head and neck squamous cell carcinoma, esophageal cancer, hepatocellular carcinoma, melanoma, sarcoma, gastric cancer, pancreatic cancer, ovarian cancer, and breast cancer.

[0138] In one implementation, cancer is selected from the group consisting of: tumors, cysts, carcinomas, sarcomas, leukemias, lymphomas, etc. For example, cancer includes, but is not limited to, mesothelioma, leukemias, and lymphomas such as cutaneous T-cell lymphoma (CTCL), non-cutaneous peripheral T-cell lymphoma, lymphomas associated with human T-cell lymphotropic virus (HTLV) such as adult T-cell leukemia / lymphoma (ATLL), B-cell lymphoma, acute non-lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, lymphoma and multiple myeloma, non-Hodgkin's lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma, Burkitt lymphoma, adult T-cell leukemia lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), or hepatocellular carcinoma. Other examples include myelodysplastic syndromes, childhood solid tumors (such as brain tumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors, and soft tissue sarcomas), common adult solid tumors (such as head and neck cancers (e.g., oral, laryngeal, nasopharyngeal, and esophageal cancers)), genitourinary cancers (e.g., prostate, bladder, kidney, uterine, ovarian, and testicular cancer), lung cancers (e.g., small cell lung cancer and non-small cell lung cancer), breast cancer, pancreatic cancer, melanoma and other skin cancers, stomach cancer, brain tumors, tumors associated with Gorlin syndrome (e.g., medulloblastoma, meningioma, etc.), and liver cancer. Additional exemplary forms of cancers treatable through the subject crystallization include, but are not limited to, bone or smooth muscle cancers, stomach cancer, small bowel cancer, rectal cancer, salivary gland cancer, endometrial cancer, adrenal cancer, anal cancer, rectal cancer, parathyroid cancer, and pituitary cancer.

[0139] The crystalline form described herein is applicable to the treatment of other cancers such as colon cancer, familial adenomatous polyposis carcinoma, and hereditary nonpolyposis colorectal cancer or melanoma. In addition, cancers include, but are not limited to, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, thyroid cancer (myeloid and papillary thyroid cancer), kidney cancer, renal parenchymal cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, testicular cancer, urethral cancer, melanoma, brain tumors (such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral neuroectodermal tumors), gallbladder cancer, bronchial cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myoma, liposarcoma, fibrosarcoma, Ewing sarcoma, and plasmacytoma.

[0140] In another aspect, this document provides for the use of one or more crystalline forms of the present disclosure in the manufacture of medicaments for treating cancer (including, but not limited to, the various types of cancer disclosed herein).

[0141] In some embodiments, the crystalline form of this disclosure can be used to treat cancers such as colorectal cancer, thyroid cancer, breast cancer, and lung cancer; and myeloproliferative disorders such as polycythemia vera, thrombocythemia, myeloid metaplasia with myelofibrosis, chronic myeloid leukemia, chronic myelomonocytic leukemia, eosinophilia syndrome, juvenile myelomonocytic leukemia, and systemic mast cell disease. In some embodiments, the crystalline form of this disclosure can be used to treat hematopoietic disorders, particularly acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute promyelocytic leukemia, and acute lymphoblastic leukemia (ALL).

[0142] Application / Dosage / Formulation

[0143] The actual dose level of the active ingredient in the pharmaceutical compositions discussed in this article can be varied in order to obtain an amount of active ingredient that is effective in achieving the desired therapeutic response for a particular patient, composition, and administration mode, without being toxic to the patient.

[0144] Specifically, the selected dose level will depend on a variety of factors, including the activity of the particular compound used, the timing of administration, the rate of excretion of the compound, the duration of treatment, other drugs, compounds or materials used in combination with the crystalline form, the age, sex, weight, condition, general health status and medical history of the patient being treated, and similar factors well known in the medical field.

[0145] A physician or veterinarian with ordinary skills in the art can readily determine and prescribe an effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian can begin administering the pharmaceutical composition at a dose level below that required to achieve the desired therapeutic effect in the crystalline form of this disclosure, and gradually increase the dose until the desired effect is achieved.

[0146] In certain implementations, it is particularly advantageous to prepare crystalline forms in dosage units for ease of application and uniform dosage.

[0147] As used herein, “unitary dosage form” refers to a physically discrete unit suitable as a single dose for a patient to be treated; each unit contains a predetermined amount of the disclosed compound bound to a desired drug medium in a manner calculated to produce the desired therapeutic effect. The crystalline unitary dosage form disclosed herein is determined by and directly depends on (a) the unique properties of the disclosed compound and the specific therapeutic effect to be achieved, and (b) the limitations inherent in the field of compounding / formulating such disclosed compounds for the treatment of patients with Kennedy's disease (SBMA).

[0148] In one embodiment, the crystalline form provided herein is formulated using one or more pharmaceutically acceptable excipients or carriers. In one embodiment, the pharmaceutical composition comprises a therapeutically effective amount of the disclosed crystalline form and a pharmaceutically acceptable carrier.

[0149] In some embodiments, the dosage of the disclosed compound is from about 1 mg to about 1000 mg. In some embodiments, the dosage of the disclosed compound used in the compositions described herein is less than about 1000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 20 mg or less than about 10 mg. For example, the dosage is about 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg or about 600 mg.

[0150] Routes of administration for any of the compositions disclosed herein include oral, nasal, rectal, vaginal, parenteral, buccal, sublingual, or topical. The compounds used herein may be formulated for administration via any suitable route, such as oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, translingual, (trans)buccal, (trans)urethral, ​​vaginal (e.g., vaginal and perivallary), nasal (internal) and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, intrathecal, subcutaneous, intramuscular, intradermal, intraarterial, intravenous, intrabronchial, inhalation, and topical administration. In one embodiment, the preferred route of administration is oral.

[0151] Suitable compositions and dosage forms include, for example, tablets, capsules, pouches, pills, soft capsules, lozenges, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, emulsions, sugar lozenges, creams, pastes, plasters, lotions, tablets, suppositories, liquid sprays for nasal or oral administration, dry powder or nebulized formulations for inhalation, and compositions and formulations for intravesical administration. It should be understood that the formulations and compositions that can be used in this disclosure are not limited to the specific formulations and compositions described herein.

[0152] For oral applications, tablets, sugar-coated pills, liquids, drops, suppositories, or capsules, pouches, and soft capsules are particularly suitable. Compositions intended for oral use can be prepared according to any method known in the art, and such compositions may contain one or more agents selected from the group consisting of inert, non-toxic pharmaceutical excipients suitable for the manufacture of tablets. Such excipients include, for example, inert diluents such as lactose; granulating and disintegrants such as corn starch; binders such as starch; and lubricants such as magnesium stearate. Tablets may be uncoated or may be coated using known techniques for aesthetic purposes or to delay the release of the active ingredient. Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert diluent.

[0153] For parenteral administration, the disclosed compounds may be formulated for injection or infusion, for example, intravenous, intramuscular, or subcutaneous injection or infusion, or for administration by bolus dose or continuous infusion. Suspensions, solutions, or emulsions in oily or aqueous media may be used, optionally containing other formulations such as suspending agents, stabilizers, or dispersants.

[0154] Those skilled in the art will recognize or be able to determine numerous equivalents to the specific procedures, implementations, claims, and embodiments described herein using only conventional experiments. Such equivalents are considered to be within the scope of this disclosure and are covered by the appended claims. For example, it should be understood that modifications to reaction conditions using art-recognized alternatives and only conventional experiments are within the scope of this application, including but not limited to reaction time, reaction scale / volume, experimental reagents (such as solvents, catalysts), pressure, atmospheric conditions (e.g., nitrogen atmosphere), and reducing / oxidizing agents.

[0155] It should be understood that whenever values ​​and ranges are provided herein, all values ​​and ranges covered by those values ​​and ranges are intended to be covered by this disclosure. Furthermore, all values ​​falling within those ranges, as well as the upper or lower limits of the ranges of values, are also covered by this application.

[0156] The following examples further illustrate various aspects of this disclosure. However, they in no way constitute a limitation on the teachings of this disclosure.

[0157] Example

[0158] This disclosure is further illustrated by the following examples, which should not be construed as further limiting. Unless otherwise indicated, practice of this disclosure will employ conventional techniques within the skill level of the art in organic synthesis, cell biology, cell culture, and molecular biology.

[0159] Synthesis of Compound 1

[0160] The synthesis of compound 1 is disclosed in international application PCT / US2022 / 021461, the entire contents of which are incorporated herein by reference. This compound is referred to as Example 239 in PCT / US2022 / 021461 and was obtained as a grayish-white solid (referred to herein as "Type A"). LCMS: m / z = 453.1 (M+H + ) 1 ¹H NMR: (400 MHz, chloroform-d) δ = 8.69–8.29 (m, 2H), 7.35 (br d, J = 7.1 Hz, 1H), 7.08 (br d, J = 7.1 Hz, 4H), 6.89–6.73 (m, 1H), 5.31 (br s, 2H), 5.00 (br s, 2H), 4.08 (br d, J = 6.5 Hz, 2H), 3.09 (br s, 3H), 1.39 (br t, J = 5.9 Hz, 3H). This compound showed an IC50 < 1 μM spectral density for NLRP3. 50 value.

[0161] Analytical methods for polymorph research

[0162] Unless otherwise stated, X-ray powder diffraction (XRPD) was performed on a Bruker D8 Advance diffractometer in reflection mode using collimated Cu Kα radiation operating at 40 kV and 40 mA. Scans were run from 2–40 degrees 2–θ in 0.02-degree steps with a scan time of 0.3 seconds per step.

[0163] Differential scanning calorimetry (DSC) was performed on a TADiscovery 2500 equipped with a Tzero disk and Tzero sealing cap, and a pinhole diameter of 0.7 mm. DSC analysis was performed from 0 °C to 250 °C at a ramp rate of 10 °C / min.

[0164] Thermogravimetric analysis (TGA) was performed on a Discovery 5500 or Q5000 at an ambient starting temperature (below 35°C) and a final temperature of 300°C (the next stage was terminated if the weight was < 80% (w / w)) with a heating rate of 10°C / min.

[0165] Nuclear magnetic resonance (NMR) analysis was performed on a Bruker Avance-AV 400M at a frequency of 400 MHz, with eight scans.

[0166] Dynamic vapor adsorption (DVS) was performed on Intrinsic, Advantage, or Adventure surfaces at an oven temperature of 25°C using water as the solvent. The sample mass was approximately 5-10 mg, and the following method was used: Cycle: 40-0-95-0-40%RH Phase step size: 10% Equilibrium: 0.002 dm / dt (% / min) Minimum dm / dt steady-state duration: 60 min Maximum dm / dt stage time: 360 min.

[0167] Example 1. Preparation of crystalline form

[0168] Compound 1, in its free form, exhibited polymorphic behavior. A total of three crystalline forms were identified as polymorphs in the free form, including one hydrate (type A) and two anhydrous forms (types B and C). Furthermore, amorphous forms were identified in hot-cold DSC cycling and variable-temperature XRPD experiments. HCl salt type A, phosphate type A, and sodium salt type A were all successfully scaled up. These scaled-up batches were the same polymorphs as those in the screening samples.

[0169] Preparation of compounds of types A, B and C

[0170] Compound of type A is a monohydrate. It can be obtained in aqueous and solvent / water systems by equilibrium and antisolvent addition. It can also be obtained in organic solvents by slow evaporation. Type A has high crystallinity. According to KF results, it contains 1.0 equivalent (3.8 wt%) of water. DSC shows a dehydration peak with an enthalpy of about 114 J / g from about 8 °C and a Tg at 134.2 °C. 起始The melting peak has an enthalpy of approximately 66 J / g. TGA showed a weight loss of approximately 3.8% at approximately 100 °C. ¹H-NMR showed no detectable residual solvent. Upon dehydration, form A is converted to the anhydrous form C. Form C is unstable and reverts to form A upon exposure to ambient conditions (approximately 23 °C, approximately 27% RH). Water activity tests showed that form A converts to form B at water activities ranging from 0 to 0.4 at 5 °C and from 0 to 0.6 at 25 °C.

[0171] Type B is an anhydrous form. It is obtained from most organic solvents through equilibrium and slow cooling. It can also be obtained from acetonitrile / MTBE and THF / MTBE by adding an antisolvent. Type B has high crystallinity. DSC shows at a TT of 174.8 °C. 起始 The melting peak has an enthalpy of approximately 117 J / g. TGA showed a weight loss of approximately 0.4% at approximately 150 °C. ¹H-NMR showed no detectable residual solvent. Water activity experiments indicated that type B is thermodynamically stable in the water activity range of 0 to 0.4 at 5 °C and in the water activity range of 0 to 0.6 at 25 °C.

[0172] Type C is an anhydrous form. It was obtained by heating Type A to 100°C under a nitrogen atmosphere in a variable-temperature XRPD experiment. Type C exhibits high crystallinity. According to the DSC thermogram of Type A, Type C shows a crystallinity at 134.2°C. 起始 It exhibits a melting peak. Type C is a metastable form. Upon exposure to environmental conditions (23°C, 27%RH), Type C transforms into Type A.

[0173] Competition equilibrium and water activity experiment of compound 1 in its free form

[0174] The relative stability of hydrate form A and anhydrous form B was investigated by competition equilibrium and water activity experiments at 5℃ and 25℃.

[0175] After 5 days of equilibration, hydrate form A was converted to anhydrous form B at water activities ranging from 0 to 0.4 at 5°C and from 0 to 0.6 at 25°C. The obtained sample remained a physical mixture of forms A and B at water activities ranging from 0.6 to 1.0 at 5°C and from 0.8 to 1.0 at 25°C. The slow form conversion kinetics are presumed to be caused by low solubility.

[0176] Based on the above results, type B is thermodynamically stable in terms of water activity in the range of 0 to 0.4 at 5°C and in the range of 0 to 0.6 at 25°C, and is considered to be the best polymorphic form in the free form.

[0177] Evaluation of Type B

[0178] Bulk stability

[0179] The volumetric stability of free form type B was evaluated over a period of one week at 25°C / 92.5%RH in an open container, at 40°C / 75%RH in an open container, and at 60°C in a closed container.

[0180] The free form, type B, is physically and chemically stable under these conditions. HPLC showed no significant degradation. XRPD showed no change in form.

[0181] solubility

[0182] The solubility of BAL-0748 in its free form, form B, was studied at 37°C for 2 h and 24 h in 2% HPMC + 1% Pluronic F68 in pH 1.2 HCl buffer, pH 4.5 50 mM acetate buffer, pH 6.8 50 mM phosphate buffer, water, pH 1.6 FaSSGF, pH 6.5 FaSSIF-v1, pH 5.0 FeSSIF-v1, and pH 4.5 50 mM acetate buffer. Residual solids after solubility tests were analyzed by XRPD.

[0183] The free form B exhibits good solubility of 0.1–0.6 mg / mL in pH 1.2 HCl buffer, pH 1.6 FaSSGF, and pH 5.0 FeSSIF-v1. In other media, the free form B shows poor solubility of 2–80 μg / mL at 2 h and 24 h.

[0184] Following solubility testing, the free form B did not show any change in form or partial conversion to the free form A in most media. However, salt formation occurred in pH 1.2 HCl buffer and pH 1.6 FaSSGF. Mono-HCl salt form A and hemi-HCl salt form A were obtained in pH 1.2 HCl buffer and pH 1.6 FaSSGF, respectively.

[0185] hygroscopic

[0186] The hygroscopicity of the free form, type B, was evaluated using a dynamic vapor adsorption (DVS) test at 25°C. The free form, type B, is slightly hygroscopic. It absorbs approximately 0.8% water at 25°C and 80% RH. After the DVS test, the resulting sample remained type B.

[0187] Preparation of HCl salt form A of compound 1

[0188] Prepare HCl salt type A using the following procedure.

[0189] 300.3 mg of compound 1 in its free form, type A, was weighed into a 20 mL glass vial. 4 mL of IPA was added to the vial at 50°C. Then, 0.57 mL of a 1.2N HCl aqueous solution (approximately 1.05 molar equivalents) was added to the above solution. After stirring at 50°C for approximately 2 minutes, a suspension was obtained.

[0190] Approximately 10 mg of HCl salt type A seed crystals were added to the above solution (suspension). After stirring at 50°C for approximately 2 hours, the solution was cooled to 25°C (suspension) by natural cooling. The suspension was then kept stirred at 25°C for approximately 5 days.

[0191] The solid was collected by centrifugation through a 0.45 μm nylon membrane filter and then dried under vacuum at 50 °C for about 2 hours. 295 mg of the HCl salt form A was obtained as a white solid in 90% yield. The characterization of compound 1, HCl salt form A, is reported in Table 9 below.

[0192] Preparation of phosphate form A of compound 1

[0193] Phosphate type A is prepared using the following procedure.

[0194] Weigh 300.0 mg of BAL-0748 in its free form, Form A, into a 20 mL glass vial. Add 7 mL of IPA to the vial at 50°C. Then add 0.48 mL of dilute phosphoric acid solution (approximately 1.05 molar equivalents, diluted with 0.9 mL of IPA to 0.1 mL of phosphoric acid) to the above solution. Obtain a suspension. Add approximately 10 mg of phosphate seed crystals, Form A, to the above solution (suspension). Stir at 50°C for approximately 2 hours, then cool the solution to 25°C by natural cooling. Keep the suspension stirred at 25°C for approximately 5 days.

[0195] The solid was collected by centrifugation and filtration, and then dried under vacuum at 50°C for about 2 hours. 336 mg of phosphate form A was obtained as a white solid in 91% yield. The characterization of compound 1 phosphate form A is reported in Table 9 below.

[0196] Preparation of sodium salt of compound 1, form A

[0197] Sodium salt type A is prepared using the following procedure.

[0198] Weigh 300.3 mg of compound 1 in its free form, form A, and approximately 29 mg of NaOH (approximately 1.05 molar equivalents) into a 20 mL glass vial. Add 3 mL of IPA to the vial at 50 °C. A suspension is obtained (almost clear solution). Add approximately 10 mg of sodium salt form A seed crystals to the above solution. After stirring at 50 °C for approximately 10 minutes, the solution gradually transforms into a suspension (almost clear solution → suspension).

[0199] After stirring at 50°C for approximately 2 hours, the solution was cooled to 25°C (suspension) by natural cooling. The suspension was kept stirred at 25°C for approximately 5 days. The solid was collected by centrifugation and filtration, and then dried under vacuum at 50°C for approximately 2 hours. 283 mg of sodium salt form A as a white solid was obtained in 89% yield. Characterization of sodium salt form A of compound 1 is reported in Table 9.

[0200] Example 2. Salt / eutectic screening

[0201] Compound 1 is a small molecule with a molecular weight of 452.46 g / mol. As predicted by Marvin Sketch v21.3, the compound contains one basic pKa of 5.16, one acidic pKa of 9.29, two H-bond donors, and five H-bond acceptors. The free form of compound 1, type A, was used as the starting material in this salt / eutectic screening study. Type A compound is a monohydrate.

[0202] Based on the pKa of compound 1, seven type I acids, two type II acids, and two type I bases were selected as salt-forming agents: hydrochloric acid, sulfuric acid, phosphoric acid, L-aspartic acid, maleic acid, L-glutamic acid, fumaric acid, methanesulfonic acid, p-toluenesulfonic acid, sodium hydroxide, and potassium hydroxide. Based on computer simulations of eutectic predictions and considering the H-bond synthon preference of compound 1, two eutectic forms were also selected to explore potential eutectics: choline and oxalic acid. IPA, EA, and acetonitrile / water (95:5, v / v) were used as screening solvents. 1.0 or 0.5 equivalents of the selected counterions / eutectic forms were applied in the screening. Slurry equilibrium, cooling, and antisolvent addition were used as crystallization methods. A total of 53 screening experiments were conducted.

[0203] Based on the salt / eutectic screening results (Table 6-8), eighteen salt / eutectic compounds and their polymorphs were identified, including HCl salt type A, hemisulfate type A, hemisulfate type B, monosulfate type A, phosphate type A, maleate type A, hemifumarate type A, hemifumarate type B, monofumarate type A, methanesulfonate type A, methanesulfonate type B, methanesulfonate type C, p-toluenesulfonate type A, p-toluenesulfonate type B, oxalate type A, sodium salt type A, potassium salt type A, and potassium salt type B. All these salt / eutectic compounds were analyzed by DSC, TGA, and other methods. 1 Further characterization was performed using ¹H-NMR, IC, KF, HPLC, and PLM.

[0204] Approximately 50 mg of compound 1 in its free form, type A, and 0.5 or 1.0 equivalents of acid or base were added to a 2 mL glass vial containing screening solvent. The resulting mixture was stirred at 50 °C for 2 hours, and then at 25 °C for approximately 3 days.

[0205] Table 6 Slurry Balance

[0206] The clarified solution obtained from the slurry equilibrium experiment was cooled to 5°C to allow solids to precipitate. After stirring at 5°C for approximately 3 days, the cooled suspension was filtered through a 0.45 µm nylon membrane filter and centrifuged at 14,000 rpm. After drying under vacuum at 50°C for 2 hours, the solids were analyzed by XRPD.

[0207] Table 7 Cooling

[0208] The clear solution or turbid suspension obtained from the cooling experiment was further processed by adding an antisolvent. The suspension obtained after adding the antisolvent was filtered through a 0.45 µm nylon membrane filter by centrifugation at 14,000 rpm. After drying under vacuum at 50 °C for 2 hours, the solids were analyzed by XRPD.

[0209] Table 8 Antisolvent Addition

[0210] Among these salt / eutectic matches, HCl salt type A, phosphate type A, and sodium salt type A exhibited favorable properties, including high purity, high crystallinity, high melting point, reasonable stoichiometry, and good counterionic safety. Therefore, they were selected as salt / eutectic candidates for scaling up and thorough evaluation in terms of volumetric stability, solubility, hygroscopicity, and morphological properties compared to those without type A.

[0211] The free form A monohydrate, HCl salt form A anhydrous, phosphate form A anhydrous, and sodium salt form A anhydrous were scaled up and fully evaluated. The results are summarized below.

[0212] Table 9 Chemical and physicochemical properties

[0213] The free form, type A, is a highly crystalline monohydrate. DSC analysis shows a dehydration peak with an enthalpy of approximately 114 J / g from about 8 °C and a Tg at 134.2 °C. 起始 The melting peak has an enthalpy of approximately 66 J / g. TGA showed a weight loss of approximately 3.8% at approximately 100 °C. HPLC showed a chemical purity of 97.8%. KF showed 3.8% water by weight, equivalent to 1.0 water molecule. 1 H-NMR showed no detectable residual solvent.

[0214] HCl salt form A is an anhydrous compound with high crystallinity. DSC analysis shows a Tc of 211.2 °C. 起始 The melting peak is visible at [location missing]. Decomposition occurs upon melting. TGA shows approximately 1.0% weight loss at 180°C. HPLC shows 98.8% chemical purity. IC [data missing] shows a stoichiometric ratio of free form to HCl of 1:0.94. 1 H-NMR showed 0.6% by weight (equivalent to 0.05 molar equivalent) of residual IPA.

[0215] Phosphate form A is an anhydrous compound with high crystallinity. DSC analysis shows that at 153.0℃... 起始 The endothermic peak at enthalpy of 2 J / g and the T at 221.1℃ 起始 The melting peak is visible at [location missing]. Decomposition occurs upon melting. TGA showed approximately 0.5% weight loss at 195°C. HPLC showed a chemical purity of 98.1%. IC [data missing] showed a stoichiometric ratio of free form to H3PO4 of 1:1.06. 1 H-NMR showed 0.2% by weight (equivalent to 0.02 molar equivalents) of residual IPA.

[0216] Sodium salt form A is a highly crystalline anhydrous compound. DSC showed no melting peak before decomposition. TGA showed approximately 1.5% weight loss at 250°C. HPLC showed a chemical purity of 98.8%. IC showed a stoichiometric ratio of free form to NaOH of 1:1.06. 1 H-NMR showed 0.4% by weight (equivalent to 0.03 molar ratio) of residual IPA.

[0217] Example 3. Volumetric Stability

[0218] The free form A, HCl salt form A, and phosphate form A are chemically and physically stable within one week under these conditions. XRPD showed no change in form. HPLC showed no significant degradation.

[0219] Sodium salt form A is chemically stable but physically unstable. HPLC showed no significant degradation. It did not show any change in form at 60°C, but dissociated into free form A after one week at 25°C / 92.5%RH and 40°C / 75%RH. This result indicates that the dissociation of the sodium salt is related to high humidity.

[0220] The results of these studies are summarized in Table 10.

[0221] Table 10 Stability: Purity and Appearance (Color, CL)

[0222] Example 4. Solubility

[0223] The solubility of free form A, HCl salt form A, phosphate form A, and sodium salt form A was measured at 37°C for 2 hours and 24 hours in 2% HPMC + 1% Pluronic F68 in pH 1.2 HCl buffer, pH 4.5 acetate buffer (50 mM), pH 6.8 phosphate buffer (50 mM), water, pH 1.6 FaSSGF, pH 6.5 FaSSIF-v1, pH 5.0 FeSSIF-v1, and pH 4.5 acetate buffer (50 mM). Residual solids after the 24-hour solubility test were analyzed by XRPD.

[0224] In pure water, HCl salt form A and phosphate form A show significantly higher solubility compared to the free form A, which should be due to pH changes.

[0225] In FeSSIF-v1 at pH 5.0, HCl salt form A showed twice the solubility of the free form A at 2 hours. At 24 hours, all three salts showed solubility comparable to the free form A.

[0226] In pH 1.2 HCl buffer, pH 4.5 acetate buffer, pH 6.8 phosphate buffer, and pH 6.5 FaSSIF-v1, the three salts showed solubility comparable to the free form, type A.

[0227] In 2% HPMC + 1% Pluronic F68 in acetate buffer (pH 4.5), sodium salt form A showed significantly improved solubility, while hydrochloride form A and phosphate form A showed solubility comparable to the free form A.

[0228] Following solubility testing, the three salt candidates partially or completely dissociated into free form, type A, in all media except pH 1.2 HCl buffer and pH 1.6 FaSSGF. Mono-HCl salt type A or hemi-HCl salt type A was obtained in pH 1.2 HCl buffer or pH 1.6 FaSSGF.

[0229] 10.4 mg of free form A, 10.0 mg of free form B, 10.8 mg of HCl salt form A, 12.2 mg of phosphate form A, or 10.5 mg of sodium salt form A were weighed into 20 mL glass vials. 5 mL of solubility medium was added. The amount of salt used was equivalent to 10 mg of anhydrous free form. The resulting suspension was stirred at 37 °C and 400 rpm, and samples were taken at 2 hours and 24 hours. The samples were centrifuged at 14,000 rpm for 5 minutes at 37 °C. The solubility and pH of the supernatant were analyzed by HPLC and pH meter, respectively. The residual solids (wet cake) from the 24-hour sample were also characterized by XRPD to determine the physical form. The results of this study are shown in Table 11.

[0230] Table 11 Solubility

[0231] Example 5. Hygroscopicity

[0232] The hygroscopicity of free form A, HCl salt form A, phosphate form A and sodium salt form A was evaluated by dynamic vapor adsorption (DVS) test at 25°C.

[0233] The free form, type A, HCl salt form A, and phosphate form A are slightly hygroscopic and showed no change in form after DVS testing at 25°C.

[0234] Sodium salt form A is moderately hygroscopic. It absorbs 2.7% water at 25°C and 80% RH. No change in form was observed after DVS testing. These results are shown in Table 12.

[0235] Table 12 Hygroscopicity

[0236] This standard is a modification of the European Pharmacopoeia's standard on hygroscopicity.

[0237] Example 6. Polymorphism

[0238] In this salt screening, one polymorph of the HCl salt (Type A); one polymorph of the phosphate salt (Type A); and one polymorph of the sodium salt (Type A) were identified. In the polymorph screening, three polymorphs in their free forms (Types A, B, and C) were obtained.

[0239] Based on the foregoing description, various modifications to this disclosure other than those described herein will be apparent to those skilled in the art. Such modifications are also intended to fall within the scope of the appended claims. Every reference cited in this application (including, but not limited to, all patents, patent applications, and publications) is incorporated herein by reference in its entirety.

Claims

1. A crystalline form of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide or a pharmaceutically acceptable salt or hydrate thereof.

2. The crystalline form as claimed in claim 1, wherein the crystalline form is a monohydrate.

3. The crystalline form as claimed in claim 1, wherein the crystalline form is anhydrous.

4. The crystalline form as described in any one of claims 1-3, wherein the crystalline form is a pharmaceutically acceptable salt.

5. The crystalline form of claim 4, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, phosphate and sodium salt.

6. The crystalline form as claimed in claim 3, wherein the crystalline form is characterized by having an XRPD diffraction pattern with peaks at angles (±0.2 degrees) of 7.2, 20.4, and 20.6 degrees, expressed in 2-θ degrees.

7. The crystalline form as claimed in claim 3 or 6, wherein the crystalline form is characterized by an XRPD diffraction pattern having peaks at angles (±0.2 degrees) of 7.2, 20.4, 20.6, and 21.6 degrees, expressed in 2-θ degrees.

8. The crystalline form as claimed in any one of claims 3, 6 and 7, wherein the crystalline form is characterized by having an XRPD diffraction pattern with peaks at angles (±0.2 degrees) of 7.2, 11.1, 11.8, 20.4, 20.6 and 21.6 degrees, expressed in 2-θ degrees.

9. The crystalline form as claimed in any one of claims 3 and 6-8, wherein the crystalline form is characterized in that the XRPD diffraction pattern has peaks at angles (±0.2 degrees) of 5.5, 7.2, 11.1, 11.8, 15.3, 20.4, 20.6, 21.6, 26.7 and 30.9 degrees, expressed in 2-θ degrees.

10. The crystalline form according to any one of claims 3 and 6-9, wherein the crystalline form has a DSC thermogram characterized by an endothermic onset temperature of 177.3°C.

11. The crystalline form as claimed in claim 2, wherein the crystalline form is characterized by having XRPD diffraction patterns with peaks at angles (±0.2 degrees) of 6.5, 12.9, and 16.1 degrees, expressed in 2-θ degrees.

12. The crystalline form as claimed in claim 2 or 11, wherein the crystalline form is characterized by an XRPD diffraction pattern having peaks at angles (±0.2 degrees) of 6.5, 11.2, 12.9, and 16.1 degrees, expressed in 2-θ degrees.

13. The crystalline form as claimed in any one of claims 2, 11, and 12, wherein the crystalline form is characterized in that the XRPD diffraction pattern has peaks at angles (±0.2 degrees) of 6.5, 11.2, 12.9, 16.1, 22.7, 24.8, and 32.6 degrees, expressed in 2-θ degrees.

14. The crystalline form as described in any one of claims 2 and 11-13, wherein the crystalline form has a DSC thermogram characterized by an endothermic onset temperature of 139.6°C.

15. The crystalline form according to any one of claims 1-5, wherein the crystalline form is 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide hydrochloride.

16. The crystalline form of claim 15, wherein the crystalline form is characterized by having an XRPD diffraction pattern with peaks at angles (±0.2 degrees) of 12.3, 18.1, and 18.2 degrees, expressed in 2-θ degrees.

17. The crystalline form as claimed in claim 15 or 16, wherein the crystalline form is characterized by an XRPD diffraction pattern having peaks at angles (±0.2 degrees) of 12.3, 12.8, 18.1, 18.2, and 28.7 degrees, expressed in 2-θ degrees.

18. The crystalline form according to any one of claims 15-17, wherein the crystalline form is characterized in that the XRPD diffraction pattern has peaks at angles (±0.2 degrees) of 12.3, 12.8, 18.1, 18.2, 26.0 and 28.7 degrees, expressed in 2-θ degrees.

19. The crystalline form according to any one of claims 15-18, wherein the crystalline form has a DSC thermogram characterized by an endothermic onset temperature of 218.3°C.

20. The crystalline form according to any one of claims 1-5, wherein the crystalline form is 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide phosphate.

21. The crystalline form of claim 20, wherein the crystalline form is characterized by having an XRPD diffraction pattern with peaks at angles (±0.2 degrees) of 3.1, 9.2, and 16.4 degrees, expressed in 2-θ degrees.

22. The crystalline form as claimed in claim 20 or 21, wherein the crystalline form is characterized by an XRPD diffraction pattern having peaks at angles (±0.2 degrees) of 3.1, 9.2, 11.7, 13.9, 15.4, 16.4, and 24.7 degrees, expressed in 2-θ degrees.

23. The crystalline form according to any one of claims 20-22, wherein the crystalline form is characterized in that the XRPD diffraction pattern has peaks at angles (±0.2 degrees) of 3.1, 9.2, 11.7, 13.9, 15.4, 16.4, 18.1, 19.0, 16.4 and 24.7 degrees, expressed in 2-θ degrees.

24. The crystalline form according to any one of claims 20-23, wherein the crystalline form has a DSC thermogram characterized by an endothermic onset temperature of 223.9°C.

25. The crystalline form according to any one of claims 1-5, wherein the crystalline form is sodium 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide.

26. The crystalline form of claim 25, wherein the crystalline form is characterized by having an XRPD diffraction pattern with peaks at angles (±0.2 degrees) of 10.3, 21.1, and 25.8 degrees, expressed in 2-θ degrees.

27. The crystalline form as claimed in claim 25 or 26, wherein the crystalline form is characterized by an XRPD diffraction pattern having peaks at angles (±0.2 degrees) of 10.3, 17.3, 20.6, 21.1, 23.3, and 25.8 degrees, expressed in 2-θ degrees.

28. The crystalline form according to any one of claims 25-27, wherein the crystalline form is characterized in that the XRPD diffraction pattern has peaks at angles (±0.2 degrees) of 10.3, 14.9, 17.2, 17.3, 18.8, 20.6, 20.0, 21.1, 23.3 and 25.8 degrees, expressed in 2-θ degrees.

29. A crystalline form of a pharmaceutically acceptable salt of 2-ethoxy-3',5'-difluoro-N-((4-(hydroxymethyl)-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-N-methyl-[1,1'-biphenyl]-4-carboxamide, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, phosphate and sodium salt.

30. A pharmaceutical composition comprising the crystalline form of any one of claims 1-29 and a pharmaceutically acceptable carrier.

31. A method for inhibiting NLRP3 inflammasomes in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of claims 1-29 or the pharmaceutical composition of claim 30.

32. A method for treating inflammation in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of claims 1-29 or the pharmaceutical composition of claim 30.

33. A method for treating inflammatory aging in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of claims 1-29 or the pharmaceutical composition of claim 30.

34. A method for treating a subject with cold pyridine-associated periodic syndrome (CAPS), the method comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of claims 1-29 or the pharmaceutical composition of claim 30.

35. A method for treating an inner ear disease or condition in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of claims 1-29 or the pharmaceutical composition of claim 30.

36. The method of claim 35, wherein the inner ear disease or condition is selected from the group consisting of: hearing loss, hearing impairment, vertigo, Meniere's disease, and tinnitus.

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