Crystalline compounds of LPAR1 antagonists

Abstract

Described herein are, inter alia, crystalline compounds of LPAR1 antagonists, pharmaceutical compositions containing the crystalline compounds, and therapeutic methods using the same.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Application No. 63 / 481,956, filed January 27, 2023, which is incorporated herein by reference in its entirety for all purposes. [Background technology]

[0002] Lysophosphatidic acid (LPA) is a family of bioactive phospholipids associated with multiple cellular functions. Although family members differ in the length and degree of saturation of their respective long-chain fatty acid backbones (Fujiwara et al., J. Biol. Chem., 2005, 280, 35038-35050), they are all capped with a glycerol-phosphate group via an ester bond. LPA is biologically produced from membrane phospholipids through a multistep cascade mediated by enzymes including lysophospholipase D (lysoPLD), autotaxin (ATX), phospholipase A1 (PLA1), phospholipase A2 (PLA2), and acylglycerol kinase (AGK) (Mutoh et al., British J. Pharmacol., 2012, 165, 829-844). Once formed, LPA can regulate numerous cell signaling pathways by binding to a class of seven membrane-domain G protein-coupled receptors (GPCRs) collectively known as LPA receptors (LPARs), six of which have been characterized as LPAR1, LPAR2, LPAR3, LPAR4, LPAR5, and LPAR6 (Choi, JW, Annu. Rev. Pharmacol. Toxicol., 2010, 50, 157-186). The biological responses elicited by LPA binding to LPARs are broad and context-dependent (Yung et al., J. Lipid Res. 2014, 55, 1192-1214; Yung et al., Neuron 2015, 85, 669-682).These may include inducing cell proliferation, stimulating cell migration and contraction, promoting axonal retraction, inhibiting apoptosis, initiating chemotaxis, closing gap junctions, and others (Chun et al., Editors, Lysophospholipid Receptors: Signaling and Biochemistry, 2013, Wiley, ISBN: 978-0-470-56905-4). Furthermore, abnormal upregulation of the LPA pathway has been implicated in multiple diseases, including cancer, inflammatory diseases, infertility, neuropathic pain, psychotic and neurodegenerative disorders, atherosclerosis, and fibrosis of the skin, kidney, lung, and liver (Choi, JW, Annu. Rev. Pharmacol. Toxicol., 2010, 50, 157-186; Noguchi et al., Curr. Opin Pharmacol., 2009, 9, 15-23; Yanagida et al., J. Biochem., 2011, 150, 223-232). As a result, targeting the LPA receptor has been and continues to be an area of ​​intense interest in identifying promising treatments for these disorders. Specifically, solutions to these and other problems in the art are disclosed herein. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Fujiwara et al.,J.Biol.Chem.,2005,280,35038-35050 [Non-patent document 2] Mutoh et al.,British J.Pharmacol.,2012,165,829-844 [Non-patent document 3] Choi, JW, Annu. Rev. Pharmacol. Toxicol., 2010, 50, 157-186 [Non-patent document 4] Yung et al.,J.Lipid Res.2014,55,1192-1214 [Non-patent document 5] Yung et al.,Neuron 2015,85,669-682 [Non-patent document 6] Chun et al.,Editors,Lysophospholipid Receptors:Signaling and Biochemistry,2013,Wiley,ISBN:978-0-470-56905-4 [Non-Patent Document 7] Noguchi et al.,Curr.Opin Pharmacol.,2009,9,15-23 [Non-patent document 8] Yanagida et al.,J.Biochem.,2011,150,223-232 Summary of the Invention

[0004] In one aspect, a crystalline compound is provided having the formula:

[0005] [ka] The crystalline compound has characteristic X-ray powder diffraction peaks at about 5.2° 2θ, about 10.4° 2θ, and about 15.6° 2θ.

[0006] In one aspect, a pharmaceutical composition is provided that includes a crystalline compound described herein and a pharmaceutically acceptable excipient.

[0007] In one aspect, a method of treating a subject in need of treatment for a neurodegenerative disorder is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0008] In one aspect, a method of treating a subject in need of treatment for an inflammatory disease is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0009] In one aspect, a method of treating a subject in need of treatment for a demyelinating disease is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0010] In one aspect, a method of treating a subject in need of treatment for a fibrotic disorder is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0011] In one aspect, a method of treating a subject in need of treatment for cancer is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound described herein.

[0012] In one aspect, a method of modulating LPAR1 activity in a subject is provided, the method comprising administering to the subject a crystalline compound described herein. [Brief explanation of the drawings]

[0013] [Figure 1] XRPD pattern of Pattern A polymorph. [Figure 2] DSC thermogram of the Pattern A polymorph. [Figure 3] TGA thermogram of Pattern A polymorph. [Figure 4] 1H-NMR spectrum of Pattern A polymorph. [Figure 5] PLM photograph of Pattern A polymorph. [Figure 6] Simulated XRPD pattern of a single crystal cultured from slow evaporation. [Figure 7] XRPD overlap of samples of the Pattern A crystalline polymorph obtained after storage at 25°C / 92% RH, 40°C / 75% RH, and 60°C for 1 week. [Figure 8A] DVS isotherm plot and DVS isotherm analysis report for the Pattern A polymorph under test conditions. [Figure 8B] DVS isotherm plot and DVS isotherm analysis report for the Pattern A polymorph under test conditions. [Figure 9] XRPD overlay of the Pattern A polymorph before (bottom) and after (top) DVS testing. [Figure 10] XRPD overlay of samples obtained by compressing the Pattern A polymorph at 5 MPa and 10 MPa. [Figure 11] XRPD overlay of samples obtained by manually dry-grinding the Pattern A polymorph with a mortar and pestle for 1 minute, 3 minutes, and 5 minutes. [Figure 12] XRPD overlay of a sample obtained by wet-milling the Pattern A polymorph with a mortar and pestle in water and ethanol. [Figure 13] XRPD overlay of samples obtained by equilibration at 25 °C for 2 weeks in methanol, ethanol, isopropanol, acetone, and methyl ethyl ketone. [Figure 14] XRPD overlay of samples obtained by equilibration in ethyl acetate, acetonitrile, IPAc, tetrahydrofuran, and 1,4-dioxane for 2 weeks at 25 °C. [Figure 15] XRPD overlay of samples obtained by equilibration in dichloromethane, 2-MeTHF, EtOH / water, acetone / water, and MEK / water for 2 weeks at 25°C. [Figure 16] XRPD overlay of samples obtained by equilibration in ACN / water, 1,4-dioxane / water, THF / water, EA / heptane, and 2-MeTHF / MTBE for 2 weeks at 25°C. [Figure 17] XRPD overlay of sample obtained by equilibration in IPA / heptane for 2 weeks at 25°C. [Figure 18] XRPD overlay of samples obtained by equilibration at 50 °C for 1 week in methanol, ethanol, isopropanol, acetone, and methyl ethyl ketone. [Figure 19] XRPD overlay of samples obtained by equilibration at 50°C for 1 week in ethyl acetate, acetonitrile, IPAc, tetrahydrofuran, and 1,4-dioxane. [Figure 20] XRPD overlay of samples obtained by equilibration at 50° C. in MeOH / water, 2-MeTHF, EtOH / water, acetone / water, and MEK / water for 1 week. [Figure 21] XRPD overlay of samples obtained by equilibration at 50° C. for 1 week in ACN / water, 1,4-dioxane / water, THF / water, EA / heptane, and 2-MeTHF / MTBE. [Figure 22] XRPD overlay of sample obtained by equilibration in IPA / heptane for 1 week at 50°C. [Figure 23] XRPD overlay of samples obtained by equilibration at 90° C. for 5 days in 1,4-dioxane and toluene. [Figure 24] XRPD overlays of samples obtained by equilibration under temperature cycling in methanol, ethanol, isopropanol, acetone, and methyl ethyl ketone. [Figure 25] XRPD overlay of samples obtained by equilibration under temperature cycling in ethyl acetate, acetonitrile, IPAc, tetrahydrofuran, and 1,4-dioxane. [Figure 26] XRPD overlays of samples obtained by equilibration under temperature cycling in MeOH / water, 2-MeTHF, EtOH / water, acetone / water, and MEK / water. [Figure 27] XRPD overlays of samples obtained by equilibration under temperature cycling in ACN / water, 1,4-dioxane / water, THF / water, EA / heptane, and 2-MeTHF / MTBE. [Figure 28] XRPD overlays of samples obtained by slow cooling in methanol, ethanol, isopropanol, acetone, and methyl ethyl ketone. [Figure 29] XRPD overlay of samples obtained by slow cooling in ethyl acetate, acetonitrile, IPAc, tetrahydrofuran, and 2-MeTHF. [Figure 30]XRPD overlays of samples obtained by flash cooling in methanol, ethanol, isopropanol, acetone, and methyl ethyl ketone. [Figure 31] XRPD overlay of samples obtained by flash quenching in ethyl acetate, acetonitrile, IPAc, tetrahydrofuran, and 2-MeTHF. [Figure 32] XRPD overlays of samples obtained by addition of antisolvent to methanol / water, methyl ethyl ketone / ACN, ethyl acetate / heptane, and tetrahydrofuran / water. [Figure 33] XRPD overlay of samples obtained by addition of anti-solvent to tetrahydrofuran / MTBE, DCM / heptane, and 2-MeTHF / heptane. [Figure 34] XRPD overlays of samples obtained by anti-solvent addition to ethanol / heptane, methyl ethyl ketone / ACN, and 2-MeTHF / ACN. [Figure 35] XRPD overlays of samples obtained by inverse addition of antisolvent to methanol / water, methyl ethyl ketone / ACN, ethyl acetate / heptane, and tetrahydrofuran / water. [Figure 36] XRPD overlay of samples obtained by reverse addition of anti-solvent to tetrahydrofuran / MTBE, DCM / heptane, and 2-MeTHF / heptane. [Figure 37] XRPD overlays of samples obtained by reverse addition of anti-solvent to ethanol / heptane, methyl ethyl ketone / ACN, and 2-MeTHF / ACN. [Figure 38] XRPD overlays of samples obtained by slow evaporation in methanol, ethanol, isopropanol, acetone, and methyl ethyl ketone. [Figure 39] XRPD overlay of samples obtained by slow evaporation in ethyl acetate, IPAc, tetrahydrofuran, 2-MeTHF, and DCM. DETAILED DESCRIPTION OF THE INVENTION

[0014] I. Definition The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulas set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0015] As used herein, the term "about" refers to a range of values ​​that includes the specified value and that one of ordinary skill in the art would consider reasonably similar to the specified value. In embodiments, about refers to within a standard deviation using generally accepted measurements in the art. In embodiments, about refers to a range that extends + / - 10% of the specified value. In embodiments, about includes the specified value. In embodiments, "about" includes the specified value (e.g., ±0.10, ±0.15, ±0.20, or ±0.25).

[0016] The term "polymorph" is used according to its ordinary meaning to refer to a crystalline form of a compound.

[0017] As used herein, the terms "crystal" or "crystalline state" or "crystalline form" refer to a physical state that is a regular three-dimensional arrangement of atoms, ions, molecules, or molecular aggregates. A crystalline state has a lattice arrangement of components arranged in a unit cell that repeats in three dimensions according to a well-defined symmetry. In contrast, the terms "amorphous" or "amorphous state" or "amorphous form" refer to a non-crystalline solid state. The physical state of a compound can be determined by techniques such as X-ray powder diffraction, polarized light microscopy, and / or differential scanning calorimetry.

[0018] The compounds, salt forms, crystalline polymorphs, therapeutic agents, or other compositions described herein may be referred to as being characterized by graphical data "substantially as shown in" a figure. Such data may include, but are not limited to, X-ray powder diffraction spectra, NMR spectra, differential scanning calorimetry curves, and thermogravimetric analysis curves, among others. As is known in the art, such graphical data may provide additional technical information to further define the compound, salt form, crystalline polymorph, therapeutic agent, or other composition. As will be appreciated by those skilled in the art, such graphical representations of data may be susceptible to small variations in peak relative intensities and peak positions due to factors such as variations in instrument response and variations in sample concentration and purity.

[0019] Substituents, when designated by their conventional chemical formula written from left to right, equally encompass the chemically identical substituents resulting from writing the structure from right to left, e.g., -CH2O- is equivalent to -OCH2-.

[0020] Unless otherwise specified, structures depicted herein are also meant to include all stereochemical forms of the structure, i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the disclosure.

[0021] "Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" refer to substances that aid in the administration and absorption of an active agent by a subject and can be included in the compositions of the present invention without causing significant adverse toxic effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline, lactated Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coating agents, sweeteners, flavoring agents, saline (such as Ringer's solution), alcohol, oils, gelatin, carbohydrates (such as lactose, amylose, or starch), fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidine, and coloring agents. Such preparations can be sterilized and, if desired, mixed with auxiliary substances such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring agents, and / or aromatic substances that do not adversely react with the compounds of the present invention. Those skilled in the art will recognize that other pharmaceutical excipients are useful in the present invention.

[0022] The term "preparation" is intended to include formulations of the active compound with an encapsulating material as a carrier to provide a capsule in which the active ingredient, with or without other carriers, is surrounded by the carrier and is therefore associated with the carrier. Also included are cachets and lozenges. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0023] As used herein, the term "administering" is used according to its plain and ordinary meaning and refers to oral administration, administration as a suppository, topical contact, intravenous, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal, or subcutaneous administration, or implantation of a sustained-release device, e.g., a mini-osmotic pump, to a subject. Administration can be by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or 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. "Coadministration" means that the compositions described herein are administered simultaneously with, immediately before, or immediately after the administration of one or more additional therapies, e.g., cancer therapies such as chemotherapy, hormonal therapy, radiation therapy, or immunotherapy. The compounds of the present invention can be administered alone or simultaneously to a patient. Co-administration is meant to include simultaneous or sequential administration of the compounds (two or more compounds) individually or in combination. Thus, the preparations can also be combined with other active agents, if desired (e.g., to reduce metabolic degradation). The compositions of the present invention can be delivered transdermally or topically, and can be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, nanoparticles, pastes, jellies, paints, powders, and aerosols.

[0024] As defined herein, the terms "inhibition," "inhibit," "inhibiting," and the like, with respect to cellular component-inhibitor interactions, mean to adversely affect (e.g., decrease) the activity or function of the cellular component (e.g., decrease a signal transduction pathway stimulated by the cellular component (e.g., a protein, ion, lipid, virus, lipid droplet, nucleic acid, nucleotide, amino acid, protein, particle, organelle, subcellular compartment, microorganism, vesicle, small molecule, protein complex, protein aggregate, or macromolecule)) compared to the activity or function of the cellular component in the absence of the inhibitor. In embodiments, inhibition means adversely affecting (e.g., decreasing) the concentration or level of the cellular component compared to the concentration or level of the cellular component in the absence of the inhibitor. In some embodiments, inhibition refers to the alleviation of a disease or disease symptom. In some embodiments, inhibition refers to a decrease in the activity of a signal transduction or signal transduction pathway (e.g., a decrease in a pathway involving the cellular component). Thus, inhibition includes, at least in part, partially or completely blocking a stimulus, reducing, preventing, or delaying activation, or inactivating a signal transduction pathway, desensitizing an enzymatically active cell, or downregulating the amount of a component.

[0025] The terms "inhibitor," "suppressor," "antagonist," or "down-regulator" refer interchangeably to a substance capable of detectably reducing the expression or activity of a given gene or protein. An antagonist can reduce expression or activity by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% compared to a control in the absence of the antagonist. In certain examples, expression or activity is 1.5-, 2-, 3-, 4-, 5-, 10-, or less fold lower than the expression or activity in the absence of the antagonist.

[0026] The term "expression" includes any step involved in the production of a polypeptide, including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification, and secretion. Expression can be detected using conventional techniques for detecting proteins (e.g., ELISA, Western blotting, flow cytometry, immunofluorescence, immunohistochemistry, etc.).

[0027] The term "lysophosphatidic acid receptor 1 antagonist" or "LPAR1 antagonist" refers to any exogenously administered compound or agent that can partially or completely inhibit or reverse the effect of an agonist (e.g., lysophosphatidic acid) on the LPAR1 receptor. This term includes compounds or agents characterized or described as antagonists, partial antagonists, and negative allosteric modulators.

[0028] Terms such as "selective" or "selectivity" with respect to a compound or agent refer to the ability of a compound or agent to preferentially increase or decrease the activity of a particular molecular target (e.g., a protein, enzyme, etc.) over one or more different molecular targets (e.g., a compound having selectivity for lysophosphatidic acid receptor 1 (LPAR1) preferentially inhibits LPAR1 over other lysophosphatidic acid receptors). In embodiments, a "lysophosphatidic acid receptor 1-selective compound" or an "LPAR1-selective compound" refers to a compound (e.g., a compound described herein) that has selectivity for lysophosphatidic acid receptor 1 (LPAR1). In embodiments, a compound (e.g., a compound described herein) is about 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, or about 100-fold selective for lysophosphatidic acid receptor 1 (LPAR1) over one or more of LPAR2, LPAR3, LPAR4, LPAR5, or LPAR6. In embodiments, a compound (e.g., a compound described herein) is at least 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, or at least 100-fold selective for lysophosphatidic acid receptor 1 (LPAR1) over one or more of LPAR2, LPAR3, LPAR4, LPAR5, or LPAR6.

[0029] A "patient" or "patient in need thereof," or a "subject" or "subject in need thereof" refers to an organism suffering from or susceptible to a disease or condition that can be treated by administration of a pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goats, sheep, cattle, deer, and other non-mammals. In an embodiment, the patient is a human. In an embodiment, the patient in need thereof is a human. In an embodiment, the subject is a human. In an embodiment, the subject in need thereof is a human.

[0030] The term "treating" or "treatment" refers to any indication of success in treatment or improvement of an injury, disease, condition, or pathology, including any objective or subjective parameter, such as relief, remission, reduction in symptoms, or making the injury, condition, or pathology more tolerable to the patient, slowing the rate of degeneration or decline, making the degenerative end point less debilitating, or improving the patient's physical or mental well-being. The treatment or improvement of symptoms can be based on objective or subjective parameters, including the results of a physical examination, neuropsychiatric examination, and / or psychiatric evaluation. For example, certain methods provided herein successfully treat cancer by reducing the incidence of cancer and / or by causing cancer remission. In some embodiments of the compositions or methods described herein, treating cancer includes slowing the rate of growth or spread of cancer cells, reducing metastasis, or slowing the growth of metastatic tumors. The term "treating" and its conjugations includes prevention of an injury, condition, pathology, or disease. In embodiments, treating is preventing. In embodiments, treating does not include preventing. In an embodiment, the treating or treatment is not a prophylactic treatment.

[0031] An "effective amount" is an amount sufficient for a compound to achieve a predetermined purpose (e.g., achieve an effect on the subject, treat a disease, reduce enzyme activity, increase enzyme activity, reduce a signal transduction pathway, or reduce one or more symptoms of a disease or condition) compared to the absence of the compound. An example of an "effective amount" is an amount sufficient to contribute to the treatment, prevention, or alleviation of one or more symptoms of a disease, which may also be referred to as a "therapeutically effective amount" when referred to in this context. "Alleviation" of one or more symptoms (and grammatical equivalents of this phrase) means a reduction in the severity or frequency of the symptoms, or the elimination of the symptoms. A "prophylactically effective amount" of a drug is an amount of a drug that, when administered to a subject, has an intended preventative effect, for example, an amount that prevents or delays the onset (or recurrence) of an injury, disease, condition, or condition, or reduces the likelihood of the onset (or recurrence) of an injury, disease, condition, or condition, or a symptom thereof. A complete preventative effect does not necessarily occur by administration of a single dose, but may occur only after administration of a series of doses. Thus, a prophylactically effective amount can be administered in one or more administrations. As used herein, an "activity-reducing amount" refers to the amount of antagonist required to reduce the activity of an enzyme compared to the absence of the antagonist. As used herein, a "function-interfering amount" refers to the amount of antagonist required to interfere with the function of an enzyme or protein compared to the absence of the antagonist. As used herein, an "activity-increasing amount" refers to the amount of agonist required to increase the activity of an enzyme compared to the absence of the agonist. As used herein, a "function-increasing amount" refers to the amount of agonist required to increase the function of an enzyme or protein compared to the absence of the agonist.The exact amount will depend on the purpose of the treatment and will be ascertainable by one of ordinary skill in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0032] "Control" or "control experiment" is used according to its plain and ordinary meaning to refer to an experiment in which the experimental subject or reagent is treated the same as in a parallel experiment except for the omission of the experimental procedure, reagent, or variable. In some cases, a control is used as a standard of comparison in evaluating experimental efficacy. In some embodiments, a control is a measurement of a protein activity (e.g., a signaling pathway) in the absence of a compound described herein (including in any embodiment, example, figure, or table).

[0033] "Disease" or "pathological condition" refers to a bodily condition or state of health of a patient or subject that can be treated with the compounds or methods provided herein. In some embodiments, the disease is a disease related to (e.g., caused by) a cellular component (e.g., a protein, ion, lipid, nucleic acid, nucleotide, amino acid, protein, particle, organelle, subcellular compartment, microorganism, vesicle, small molecule, protein complex, protein aggregate, or macromolecule). In embodiments, the disease is a neurodegenerative disease. In embodiments, the disease is an inflammatory disease. In embodiments, the disease is post-hemorrhagic encephalitis. In embodiments, the disease is a demyelinating disease. In embodiments, the disease is multiple sclerosis. In embodiments, the disease is a fibrotic disease. In embodiments, the disease is pulmonary fibrosis. In embodiments, the disease is idiopathic pulmonary fibrosis. In embodiments, the disease is cancer. In embodiments, the disease is glioblastoma.

[0034] As used herein, the term "neurodegenerative disease" or "neurodegenerative disorder" refers to a disease or condition in which the function of a subject's nervous system is impaired. Examples of neurodegenerative diseases that may be treated with the compounds, pharmaceutical compositions, or methods described herein include Alexander disease, Alper's disease, Alzheimer's disease, amyotrophic lateral sclerosis, ataxia-telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Batten disease), bovine spongiform encephalopathy, and cerebrospinal fluid disorders (BSA). encephalopathy, BSE), Canavan disease, Cockayne syndrome, corticobasal degeneration, Creutzfeldt-Jakob disease, frontotemporal dementia, Gerstmann-Straussler-Scheinker syndrome, Huntington's disease, HIV-associated dementia, Kennedy disease, Krave disease, Kuhl, dementia with Lewy bodies, Machado-Joseph disease (Spinocerebellar ataxia type 3), multiple sclerosis, multiple system atrophy, narcolepsy, neuroborreliosis, Parkinson's disease, Pelizaeus-Merzbach disease, Pick's disease, primary lateral sclerosis, prion diseases, Refsum disease, Sandhoff disease, Schilder's disease, subacute combined spinal degeneration secondary to pernicious anemia, schizophrenia, spinocerebellar ataxia (various types with varying features), spinal muscular atrophy, Steele-Richardson-Olszewski disease, or tabes dorsalis.

[0035] As used herein, the term "inflammatory disease" refers to a disease or condition characterized by abnormal inflammation (e.g., increased levels of inflammation compared to a control, such as a healthy individual without the disease). Examples of inflammatory diseases include autoimmune diseases, arthritis, rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, multiple sclerosis, systemic lupus erythematosus, and the like. erythematosus, SLE), myasthenia gravis, juvenile-onset diabetes mellitus, type 1 diabetes mellitus, Guillain-Barré syndrome, Hashimoto's encephalopathy, Hashimoto's thyroiditis, ankylosing spondylitis, psoriasis, Sjögren's syndrome, vasculitis, glomerulonephritis, autoimmune thyroiditis, Behçet's disease, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, ichthyosis, Graves' ophthalmopathy, inflammatory bowel disease, Addison's disease, vitiligo, asthma, allergic asthma, acne vulgaris, celiac disease, chronic prostatitis, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, sarcoidosis, transplant rejection, interstitial cystitis, atherosclerosis, scleroderma, and atopic dermatitis.

[0036] As used herein, the term "demyelinating disease" refers to any disease or condition characterized by damage to the protective covering (e.g., myelin sheath) surrounding nerve fibers (e.g., in the brain, optic nerve, or spinal cord). In embodiments, the demyelinating disease is a demyelinating disease of the central nervous system. In embodiments, the demyelinating disease is multiple sclerosis. In embodiments, the demyelinating disease is a demyelinating disease of the peripheral nervous system.

[0037] As used herein, the terms "fibrotic disease" and "fibrosis" refer to any disease or condition characterized by the formation of excess fibrous connective tissue. The formation of excess fibrous connective tissue may be responsive to a repair or reactive process. Fibrotic diseases include, but are not limited to, pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis (IPF)), liver fibrosis (e.g., nonalcoholic steatohepatitis (NASH)), bone marrow fibrosis, skin fibrosis (e.g., scleroderma), ocular fibrosis, mediastinal fibrosis, cardiac fibrosis, renal fibrosis, interstitial fibrosis, epidural fibrosis, epithelial fibrosis, or idiopathic fibrosis.

[0038] As used herein, the term "cardiovascular disorder" or "cardiovascular disease" is used according to its plain and ordinary meaning. In embodiments, cardiovascular diseases that may be treated with the compounds, pharmaceutical compositions, or methods described herein include, but are not limited to, stroke, heart failure, hypertension, hypertensive heart disease, myocardial infarction, angina pectoris, tachycardia, cardiomyopathy, rheumatic heart disease, cardiomyopathies, cardiac arrhythmias, congenital heart disease, valvular heart disease, carditis, aortic aneurysm, peripheral arterial disease, thromboembolism, and venous thrombosis.

[0039] As used herein, the term "cancer" refers to all types of cancers, neoplasms, or malignant tumors found in mammals (e.g., humans), including leukemia, lymphoma, carcinoma, and sarcoma. Exemplary cancers that may be treated with the compounds or methods provided herein include thyroid cancer, endocrine system cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, melanoma, mesothelioma, ovarian cancer, sarcoma, gastric cancer, uterine cancer, medulloblastoma, colorectal cancer, or pancreatic cancer. Further examples may include Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primary thrombocythemia, primary macroglobulinemia, primary brain tumor, cancer, malignant pancreatic insulanoma, malignant carcinoid, bladder cancer, premalignant skin lesions, testicular cancer, lymphoma, thyroid cancer, esophageal cancer, genitourinary cancer, malignant hypercalcemia, endometrial cancer, adrenocortical carcinoma, endocrine or exocrine pancreatic neoplasms, medullary thyroid cancer, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, or prostate cancer.

[0040] The term "leukemia" refers broadly to progressive, malignant diseases of the blood-forming organs, generally characterized by distorted growth and development of white blood cells and their precursor cells in the blood and bone marrow. Leukemias are generally classified clinically based on (1) the duration and character of the disease—acute or chronic, (2) the type of cell involved—bone marrow (myeloid), lymphatic (lymphatic), or monocytic, and (3) the increased or non-increasing number of abnormal cells in the blood—leukemia or non-leukemia (subleukemia). Exemplary leukemias that may be treated with the compounds or methods provided herein include, for example, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, non-leukemic leukemia, leukemic leukemia, basophilic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross leukemia, hairy cell leukemia, hemoblastic leukemia, hemoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia leukopenic leukemia, lymphocytic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myeloblastic leukemia, myelogranulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia, multiple myeloma, plasma cell leukemia, promyelocytic leukemia, Leder cell leukemia, Schilling leukemia, stem cell leukemia, subleukemic leukemia, or undifferentiated cell leukemia.

[0041] As used herein, the term "lymphoma" refers to a group of cancers affecting hematopoietic and lymphatic tissues. It develops primarily in lymphocytes, which are blood cells found in lymph nodes, spleen, thymus, and bone marrow. The two main types of lymphoma are non-Hodgkin's lymphoma and Hodgkin's disease. Hodgkin's disease accounts for approximately 15% of all diagnosed lymphomas. It is a cancer associated with Reed-Sternberg malignant B lymphocytes. Non-Hodgkin's lymphoma (NHL) can be classified based on the rate at which the cancer grows and the type of cell involved. Types of NHL include aggressive (high-grade) and indolent (low-grade). Based on the type of cell involved, there are B-cell and T-cell NHL. Exemplary B-cell lymphomas that may be treated with the compounds or methods provided herein include, but are not limited to, small lymphocytic lymphoma, mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, extranodal (MALT) lymphoma, nodal (monocytic B-cell) lymphoma, splenic lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, lymphoblastic lymphoma, immunoblastic large cell lymphoma, or precursor B-lymphoblastic lymphoma. Exemplary T-cell lymphomas that may be treated with the compounds or methods provided herein include, but are not limited to, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, anaplastic large cell lymphoma, mycosis fungoides, and precursor T-lymphoblastic lymphoma.

[0042] The term "sarcoma" generally refers to a tumor composed of closely packed cells that are composed of a substance like embryonic connective tissue and that is generally composed of tightly packed cells embedded in a fibrous or homogeneous substance. Sarcomas that may be treated with the compounds or methods provided herein include chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abemethy's sarcoma, liposarcoma, liposarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma, green sarcoma, choriocarcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblastic sarcoma, and fibroblastic sarcoma. These include alveolar sarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, B-cell immunoblastic sarcoma, lymphoma, T-cell immunoblastic sarcoma, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukemia sarcoma, malignant mesenchymal sarcoma, parosteal sarcoma, reticulocytic sarcoma, Rous sarcoma, serous cystic sarcoma, synovial sarcoma, and telangiectatic sarcoma.

[0043] The term "melanoma" is intended to mean a tumor arising from the melanocytic system of the skin and other organs. Melanomas that may be treated with the compounds or methods provided herein include, for example, acral lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, nodular melanoma, subungual melanoma, or superficial spreading melanoma.

[0044] The term "carcinoma" refers to a malignant new growth composed of epithelial cells that tend to infiltrate surrounding tissues and give rise to metastases. Exemplary carcinomas that may be treated with the compounds or methods provided herein include, for example, medullary thyroid carcinoma, familial medullary thyroid carcinoma, lobular cell carcinoma, acinic cell carcinoma, adenocell carcinoma, adenoid cystic carcinoma, carcinoma adenomatous, adrenocortical carcinoma, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid cell carcinoma, basosquamous cell carcinoma, bronchioloalveolar carcinoma, bronchiolocarcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocarcinoma, choriocarcinoma, colloid carcinoma, comedocarcinoma, corpus carcinoma, cribriform carcinoma, armored carcinoma, skin carcinoma, cylindrical carcinoma, cylindrical cell carcinoma, ductal carcinoma, and compact carcinoma. durum, embryonal carcinoma, encephalomyocellular carcinoma, epidermoid carcinoma, epidermoid carcinoma, tonsillar carcinoma, exophytic carcinoma, ulcer carcinoma, fibrous carcinoma, gelatinous carcinoma, giant cell carcinoma, adenocarcinoma, granulosa cell carcinoma, hair matrix carcinoma, hematoid carcinoma, hepatocellular carcinoma, Hürthle cell carcinoma, glassy carcinoma, adrenal-like carcinoma, infantile embryonal carcinoma, carcinoma in situ, intraepidermal carcinoma, intraepithelial carcinoma, Krompecher carcinoma, Kulchitzky cell carcinoma, large cell carcinoma, lenticular carcinoma, lipomatous carcinoma, lymphoepithelial carcinoma, medullary carcinoma, melanoma, carcinoma molle, mucinous carcinoma carcinoma, mucinous carcinoma (carcinoma muciparum), mucous cell carcinoma, mucoepidermoid carcinoma, mucinous carcinomamucosum, mucosal carcinoma, myxomatous carcinoma, nasopharyngeal carcinoma, oat cell carcinoma, ossifying carcinoma, osteoid carcinoma, papillary carcinoma, periportal carcinoma, preinvasive carcinoma, squamous cell carcinoma, pasty carcinoma, renal cell carcinoma of the kidney, storage cell carcinoma, sarcomatoid carcinoma, Schneiderian carcinoma, scirrhous carcinoma, scrotal carcinoma, signet ring cell carcinoma, simplex carcinoma, small cell carcinoma, solanoid carcinoma, spheroid cell carcinoma, spindle cell carcinoma, porotic carcinoma, squamous cell carcinoma, squamous cell carcinoma, string carcinoma, telangiectatic carcinoma, telangiectatic-like carcinoma, transitional cell carcinoma, nodular carcinoma (carcinoma tuberosum), nodular carcinoma, verrucous carcinoma, or choriocarcinoma.

[0045] As used herein, the terms "metastasis," "metastatic," and "metastatic cancer" can be used interchangeably and refer to the spread of a proliferative disease or disorder, e.g., cancer, from one organ or another non-adjacent organ or part of the body. "Metastatic cancer" is also referred to as "stage IV cancer." Cancer begins at a site of origin, e.g., the breast, referred to as a primary tumor, e.g., primary breast cancer. Some cancer cells in the primary tumor or site of origin acquire the ability to penetrate and invade surrounding normal tissue in the local area and / or penetrate the walls of the lymphatic or vascular system and circulate through the system to other parts and tissues of the body. Clinically detectable secondary tumors formed from cancer cells of the primary tumor are referred to as metastatic or secondary tumors. When cancer cells metastasize, the metastatic tumor and its cells are presumed to be similar to those of the original tumor. Thus, if lung cancer metastasizes to the breast, the secondary tumor in the breast site will consist of abnormal lung cells, not abnormal breast cells. The secondary tumor in the breast is referred to as metastatic lung cancer. Therefore, the term metastatic cancer refers to a disease in which a subject has or has had a primary tumor, and has one or more secondary tumors.The term non-metastatic cancer or a subject with non-metastatic cancer refers to a disease in which a subject has a primary tumor but does not have one or more secondary tumors.For example, metastatic lung cancer refers to a disease in a subject who has or has had a history of a primary lung tumor, and has one or more secondary tumors, for example, in a second location or multiple locations in the breast.

[0046] The terms "cutaneous metastasis" or "skin metastasis" refer to secondary malignant cell growth in the skin, where the malignant cells originate from the primary cancer site (e.g., breast). In cutaneous metastasis, cancerous cells from the primary cancer site migrate to the skin, where they may divide and cause lesions. Skin metastasis can result from the migration of cancer cells from a breast cancer tumor to the skin.

[0047] The term "visceral metastasis" refers to secondary malignant cell growth in an internal organ (e.g., heart, lung, liver, pancreas, intestine) or body cavity (e.g., pleura, peritoneum), where the malignant cells originate from the primary cancer site (e.g., head and neck, liver, breast). In visceral metastasis, cancerous cells from the primary cancer site migrate to an internal organ, where they may divide and cause lesions. Visceral metastasis can result from the migration of cancer cells from a liver cancer tumor or a head and neck tumor to an internal organ.

[0048] As used herein, the term "LPAR1-associated disease" refers to any disease or condition caused by aberrant activity or signaling of LPAR1. In embodiments, the LPAR1-associated disease is a neurodegenerative disease. In embodiments, the LPAR1-associated disease is an inflammatory disease. In embodiments, the LPAR1-associated disease is post-hemorrhagic encephalitis. In embodiments, the LPAR1-associated disease is a demyelinating disease. In embodiments, the LPAR1-associated disease is multiple sclerosis. In embodiments, the LPAR1-associated disease is a fibrotic disease. In embodiments, the LPAR1-associated disease is pulmonary fibrosis. In embodiments, the LPAR1-associated disease is idiopathic pulmonary fibrosis. In embodiments, the LPAR1-associated disease is cancer. In embodiments, the LPAR1-associated disease is glioblastoma.

[0049] The term "drug" is used according to its ordinary meaning to refer to a substance that has a physiological effect (e.g., a beneficial effect, useful for treating a subject) when introduced into or to a subject (e.g., into or on the body of a subject or patient). The drug moiety is the radical of the drug.

[0050] In therapeutic use for the treatment of disease, the compounds utilized in the pharmaceutical compositions of the present invention may be administered at an initial dosage of about 0.001 mg / kg to about 1000 mg / kg daily. Daily dosage ranges of about 0.01 mg / kg to about 500 mg / kg, or about 0.1 mg / kg to about 200 mg / kg, or about 1 mg / kg to about 100 mg / kg, or about 10 mg / kg to about 50 mg / kg may be used. However, dosages may vary depending on the requirements of the patient, the severity of the condition being treated, and the compound or drug being used. For example, dosages may be empirically determined taking into account the type and stage of disease (e.g., multiple sclerosis, fibrotic disease, encephalitis, or cancer) diagnosed in a particular patient. In the context of the present invention, the dose administered to a patient should be sufficient to effect a beneficial therapeutic response in the patient over time. The size of the dose will also be determined by the existence, nature, and extent of any adverse side effects associated with the administration of the compound in a particular patient. The determination of suitable dosage for specific situation is within the skill of a specialist.Generally, treatment is started with a lower dosage that is less than the optimal dose of compound.Then, dosage is gradually increased until the optimal effect is reached under the circumstances.For convenience, if desired, total daily dosage can be divided and administered in portions throughout the day.

[0051] In the context of a substance or substance activity or function related to a disease (e.g., a protein-related disease, a disease associated with a cellular component), the term "related to" or "associated with" means that the substance or the substance activity or function causes (in whole or in part) the disease (e.g., multiple sclerosis, a fibrotic disease, encephalitis, or cancer) or causes (in whole or in part) the symptoms of the disease, or the symptoms of the disease can be treated by modulating (e.g., inhibiting or activating) the substance (e.g., a cellular component). As used herein, something that is described as associated with a disease, if it is a causative agent, can be a target for treating the disease.

[0052] As used herein, the term "aberrant" refers to something that is different from normal. When used to describe enzyme activity, abnormal refers to activity that is greater than or less than the average of a normal control or normal, non-disease control sample. Abnormal activity can refer to an amount of activity that results in disease, and restoring the abnormal activity to a normal or non-disease-associated amount (e.g., by administering a compound or using the methods described herein) results in a reduction of the disease or one or more disease symptoms.

[0053] The term "lysophosphatidic acid receptor" or "LPAR" refers to one or more of a family of G protein-coupled receptors for lysophosphatidic acid (LPA). In embodiments, LPARs include LPAR1, LPAR2, LPAR3, LPAR4, LPAR5, and LPAR6.

[0054] The term "lysophosphatidic acid receptor 1" or "LPAR1" refers to a G protein-coupled receptor (including homologs, isoforms, and functional fragments thereof) that binds the lipid signaling molecule lysophosphatidic acid (LPA). This term includes any recombinant or native form of an LPAR1 variant thereof that maintains LPAR1 activity (e.g., within the range of at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% activity compared to wild-type LPAR1). In embodiments, the LPAR1 protein encoded by the LPAR1 gene has an amino acid sequence set forth in or corresponding to Entrez 1902, UniProt Q92633, RefSeq(protein) NP_001392.2, or RefSeq(protein) NP_476500.1. In an embodiment, the LPAR1 gene has the nucleic acid sequence set forth in RefSeq(mRNA)NM_001401.3 or RefSeq(mRNA)NM_057159.2. In an embodiment, the amino acid sequence or nucleic acid sequence is a sequence known at the time of filing this application.

[0055] II. Compounds In one aspect, there is provided a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the following structure:

[0056] [ka]

[0057] In one aspect, a crystalline compound is provided having the formula:

[0058] [ka] The crystalline compound has characteristic X-ray powder diffraction peaks at about 5.2° 2θ, about 10.4° 2θ, and about 15.6° 2θ.

[0059] In embodiments, the crystalline compound has additional characteristic X-ray powder diffraction peaks at about 11.3°2θ, about 12.6°2θ, about 18.4°2θ, about 20.9°2θ, about 22.4°2θ, about 26.1°2θ, about 27.4°2θ, and about 27.6°2θ.

[0060] In embodiments, the crystalline compound is oriented at about 6.7°2θ, about 7.3°2θ, about 8.6°2θ, about 9.4°2θ, about 11.3°2θ, about 12.0°2θ, about 12.6°2θ, about 13.6°2θ, about 14.9°2θ, about 16.8°2θ, about 17.4°2θ, about 18.4°2θ, about 18.8°2θ, about 19.5°2θ, about 20.1°2θ, about 20.9°2θ, about 21.8°2θ, about 22.4°2θ, about 22.7°2θ, about 23.1°2θ, about 24.1°2θ, about 24.4°2θ, about 24.9°2θ, about and about 39.6°2θ.

[0061] The peaks in the X-ray powder diffraction pattern of the crystalline compound are shown in Table 1.

[0062] [Table 1]

[0063] In embodiments, the X-ray powder diffraction pattern of the crystalline compound has one or more peaks set forth in Table 1.

[0064] In an embodiment, the crystalline compound is further characterized by having a differential scanning calorimetry endotherm onset at about 188.6°C.

[0065] In one aspect, a compound is provided having the formula:

[0066] [ka] The compound is in a crystalline form, which has characteristic X-ray powder diffraction peaks at about 5.2° 2θ, about 10.4° 2θ, and about 15.6° 2θ.

[0067] In embodiments, the crystalline form has additional characteristic X-ray powder diffraction peaks at about 11.3°2θ, about 12.6°2θ, about 18.4°2θ, about 20.9°2θ, about 22.4°2θ, about 26.1°2θ, about 27.4°2θ, and about 27.6°2θ.

[0068] In embodiments, the crystalline form is selected from the group consisting of about 6.7 °2Θ, about 7.3 °2Θ, about 8.6 °2Θ, about 9.4 °2Θ, about 11.3 °2Θ, about 12.0 °2Θ, about 12.6 °2Θ, about 13.6 °2Θ, about 14.9 °2Θ, about 16.8 °2Θ, about 17.4 °2Θ, about 18.4 °2Θ, about 18.8 °2Θ, about 19.5 °2Θ, about 20.1 °2Θ, about 20.9 °2Θ, about 21.8 °2Θ, about 22.4 °2Θ, about 22.7 °2Θ, about 23.1 °2Θ, about 24.1 °2Θ, about 24.4 °2Θ, about 24.9 °2Θ, about 2 and about 39.6°2θ.

[0069] III. Method for producing crystalline compounds In one aspect, a method for producing a crystalline compound is provided, the method comprising: (i) a solvent component and a solvent of formula (I):

[0070] [ka] and a compound having the formula: (ii) obtaining a crystalline compound.

[0071] In embodiments, the solvent component comprises one or more selected from the group consisting of methanol, ethanol, isopropanol, acetone, methyl ethyl ketone, ethyl acetate, acetonitrile, isopropyl acetate, tetrahydrofuran, 1,4-dioxane, dichloromethane, 2-methyltetrahydrofuran, water, heptane, and methyl tert-butyl ether. In embodiments, the solvent component is methanol. In embodiments, the solvent component is ethanol. In embodiments, the solvent component is isopropanol. In embodiments, the solvent component is acetone. In embodiments, the solvent component is methyl ethyl ketone. In embodiments, the solvent component is ethyl acetate. In embodiments, the solvent component is acetonitrile. In embodiments, the solvent component is isopropyl acetate. In embodiments, the solvent component is tetrahydrofuran. In embodiments, the solvent component is 1,4-dioxane. In embodiments, the solvent component is dichloromethane. In embodiments, the solvent component is 2-methyltetrahydrofuran. In embodiments, the solvent component is a mixture of ethanol and water. In embodiments, the solvent component is a mixture of methyl ethyl ketone and water. In embodiments, the solvent component is a mixture of acetonitrile and water. In embodiments, the solvent component is a mixture of 1,4-dioxane and water. In embodiments, the solvent component is a mixture of tetrahydrofuran and water. In embodiments, the solvent component is a mixture of ethyl acetate and heptane. In embodiments, the solvent component is a mixture of 2-methyltetrahydrofuran and methyl tert-butyl ether. In embodiments, the solvent component is a mixture of isopropanol and heptane.

[0072] In an embodiment, the solvent component and compound are mixed at 25°C for two weeks. In an embodiment, the solvent component and compound are mixed at 50°C for one week. In an embodiment, the solvent component and compound are mixed at 90°C for five days. In an embodiment, the solvent component and compound are mixed under a temperature cycle of 5°C to 50°C at a cooling rate of 0.1°C per minute for 10 cycles.

[0073] In embodiments, step (ii) comprises filtering the mixture. In embodiments, step (ii) comprises filtering the mixture by centrifugation. In embodiments, step (ii) comprises filtering the mixture through a 0.45 μm nylon membrane filter by centrifugation. In embodiments, step (ii) comprises filtering the mixture through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm.

[0074] In an embodiment, step (ii) comprises slow evaporation of the solvent component.

[0075] In one aspect, a method for producing a crystalline compound is provided, the method comprising: (i) a solvent component and a solvent of formula (I):

[0076] [ka] and a compound having the formula: (ii) heating the mixture to a first temperature; (iii) filtering the mixture to obtain a filtrate; (iv) cooling the filtrate to a second temperature to obtain a precipitate; (v) filtering the precipitate to obtain a crystalline compound.

[0077] In an embodiment, the first temperature range is about 40°C to about 60°C. In an embodiment, the first temperature is about 40°C. In an embodiment, the first temperature is about 45°C. In an embodiment, the first temperature is about 50°C. In an embodiment, the first temperature is about 55°C. In an embodiment, the first temperature is about 60°C.

[0078] In an embodiment, the second temperature range is about 0°C to about 10°C. In an embodiment, the second temperature is about 0°C. In an embodiment, the second temperature is about 5°C. In an embodiment, the first temperature is about 10°C.

[0079] In an embodiment, the method further comprises, after step (iv), further cooling the mixture to about -20°C.

[0080] In embodiments, the solvent component comprises one or more selected from the group consisting of methanol, ethanol, isopropanol, acetone, methyl ethyl ketone, ethyl acetate, acetonitrile, isopropyl acetate, tetrahydrofuran, and 2-methyltetrahydrofuran. In embodiments, the solvent component is methanol. In embodiments, the solvent component is ethanol. In embodiments, the solvent component is isopropanol. In embodiments, the solvent component is acetone. In embodiments, the solvent component is methyl ethyl ketone. In embodiments, the solvent component is ethyl acetate. In embodiments, the solvent component is acetonitrile. In embodiments, the solvent component is isopropyl acetate. In embodiments, the solvent component is tetrahydrofuran. In embodiments, the solvent component is 2-methyltetrahydrofuran.

[0081] In one aspect, a method for producing a crystalline compound is provided, the method comprising: (i) a solvent component and a solvent of formula (I):

[0082] [ka] and a compound having the formula: (ii) filtering the mixture to obtain a filtrate; (iii) adding a poor solvent component to the filtrate to obtain a precipitate; (iv) filtering the precipitate to obtain a crystalline compound.

[0083] In an embodiment, the method further comprises, after step (iii), cooling the mixture to about -20°C.

[0084] In embodiments, the solvent component comprises one or more selected from the group consisting of methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, tetrahydrofuran, 1,4-dioxane, dichloromethane, and 2-methyltetrahydrofuran. In embodiments, the solvent component is methanol. In embodiments, the solvent component is ethanol. In embodiments, the solvent component is acetone. In embodiments, the solvent component is methyl ethyl ketone. In embodiments, the solvent component is ethyl acetate. In embodiments, the solvent component is tetrahydrofuran. In embodiments, the solvent component is 1,4-dioxane. In embodiments, the solvent component is dichloromethane. In embodiments, the solvent component is 2-methyltetrahydrofuran.

[0085] In embodiments, the anti-solvent component comprises one or more selected from the group consisting of water, heptane, methyl tert-butyl ether, and acetonitrile. In embodiments, the anti-solvent component is water. In embodiments, the anti-solvent component is heptane. In embodiments, the anti-solvent component is methyl tert-butyl ether. In embodiments, the anti-solvent component is acetonitrile.

[0086] In one aspect, a method for producing a crystalline compound is provided, the method comprising: (i) a solvent component and a solvent of formula (I):

[0087] [ka] and a compound having the formula: (ii) filtering the mixture to obtain a filtrate; (iii) adding the filtrate to a poor solvent component to obtain a precipitate; (iv) filtering the precipitate to obtain a crystalline compound.

[0088] In an embodiment, step (iii) comprises rapidly adding the filtrate to the anti-solvent component.

[0089] In an embodiment, the method further comprises, after step (iii), cooling the mixture to about -20°C.

[0090] In embodiments, the solvent component comprises one or more selected from the group consisting of methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, tetrahydrofuran, 1,4-dioxane, dichloromethane, and 2-methyltetrahydrofuran. In embodiments, the solvent component is methanol. In embodiments, the solvent component is ethanol. In embodiments, the solvent component is acetone. In embodiments, the solvent component is methyl ethyl ketone. In embodiments, the solvent component is ethyl acetate. In embodiments, the solvent component is tetrahydrofuran. In embodiments, the solvent component is 1,4-dioxane. In embodiments, the solvent component is dichloromethane. In embodiments, the solvent component is 2-methyltetrahydrofuran.

[0091] In embodiments, the anti-solvent component comprises one or more selected from the group consisting of water, heptane, methyl tert-butyl ether, and acetonitrile. In embodiments, the anti-solvent component is water. In embodiments, the anti-solvent component is heptane. In embodiments, the anti-solvent component is methyl tert-butyl ether. In embodiments, the anti-solvent component is acetonitrile.

[0092] IV. Pharmaceutical Compositions In one aspect, a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the following structure:

[0093] [ka] and a pharmaceutically acceptable excipient.

[0094] In one aspect, a pharmaceutical composition is provided that includes a crystalline compound described herein and a pharmaceutically acceptable excipient.

[0095] In embodiments, the pharmaceutical composition does not include any crystalline form of the compound other than the crystalline compounds described herein, hi embodiments, the pharmaceutical composition does not include any amorphous form of the compound.

[0096] In embodiments, the pharmaceutical compositions do not include crystalline forms of the compound other than those described herein.

[0097] In embodiments, the pharmaceutical composition comprises an effective amount of a crystalline compound described herein. In embodiments, the pharmaceutical composition comprises a therapeutically effective amount of a crystalline compound described herein.

[0098] In embodiments, the crystalline compound is further processed to provide a more uniform particle size, to control particle size, or to reduce particle size. For example, the initial crystalline material may be subjected to mechanical impact means such as crushing, grinding, milling (e.g., ball milling and jet milling) to provide particles having a desired particle size distribution.

[0099] In embodiments, the crystalline compounds described herein are substantially pure in that they contain less than about 5%, or less than about 1%, or less than about 0.1% of other small organic molecules, such as contaminating intermediates or by-products produced in one or more steps of the synthetic method. In embodiments, the crystalline compounds described herein are substantially pure in that they contain less than about 5% of other small organic molecules, such as contaminating intermediates or by-products produced in one or more steps of the synthetic method. In embodiments, the crystalline compounds described herein are substantially pure in that they contain less than about 1% of other small organic molecules, such as contaminating intermediates or by-products produced in one or more steps of the synthetic method. In embodiments, the crystalline compounds described herein are substantially pure in that they contain less than about 0.1% of other small organic molecules, such as contaminating intermediates or by-products produced in one or more steps of the synthetic method.

[0100] These pharmaceutical compositions include those suitable for oral, rectal, topical, buccal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), vaginal, ophthalmic, or aerosol administration.

[0101] Exemplary pharmaceutical compositions are used in the form of pharmaceutical preparations, e.g., in solid, semi-solid, or liquid form, containing the crystalline compound as an active ingredient in a mixture with an organic or inorganic carrier or excipient suitable for external, enteral, or parenteral administration. In embodiments, the crystalline compound is formulated with conventional non-toxic pharmaceutically acceptable carriers for, e.g., tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions, and any other form suitable for use. The crystalline compound is included in the pharmaceutical composition in an amount sufficient to produce the desired effect on the disease process or condition.

[0102] In embodiments for preparing solid compositions such as tablets, the crystalline compound is mixed with a pharmaceutical carrier, e.g., conventional tablet ingredients such as cornstarch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, or gums, and other pharmaceutical diluents, e.g., water, to form a solid preformulation composition containing a homogeneous mixture of the crystalline compound described herein. When these preformulation compositions are referred to as homogeneous, it is meant that the crystalline compound is dispersed evenly throughout the composition such that the composition is easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules.

[0103] In solid dosage forms for oral administration (e.g., capsules, tablets, pills, dragees, powders, granules, etc.), the crystalline compound is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, hypromellose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as crospovidone, croscarmellose sodium, sodium starch glycolate, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) dissolution retarders, such as paraffin; and (6) absorption enhancers, such as quaternary ammonium compounds. (7) Wetting agents such as docusate sodium, cetyl alcohol, and glycerol monostearate. (8) Absorbents such as kaolin and bentonite clay. (9) Lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. (10) Coloring agents. For capsules, tablets, and pills, in some embodiments, the compositions also comprise buffering agents. In some embodiments, solid compositions of a similar type are also employed as fillers for soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycols.

[0104] In embodiments, tablets are made by compression or molding, optionally with one or more accessory ingredients. In embodiments, compressed tablets are prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents, or dispersing agents. In embodiments, molded tablets are made by molding in a suitable machine a mixture of the crystalline compound moistened with an inert liquid diluent. In embodiments, tablets and other solid dosage forms, such as dragees, capsules, pills, and granules, are scored or prepared with coatings and shells, such as enteric coatings and other coatings.

[0105] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the crystalline compound, in some embodiments, the liquid dosage form contains an inert diluent, such as water or other solvent, a solubilizer and emulsifier, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, cyclodextrins, and mixtures thereof.

[0106] In embodiments, the suspension comprises a suspending agent in addition to the crystalline compound, such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.

[0107] In embodiments, formulations for rectal or vaginal administration are presented as suppositories, which are prepared by mixing the crystalline compound with one or more suitable non-irritating excipients or carriers, including, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which are solid at room temperature but liquid at body temperature, and therefore melt within the body cavity to release the active agent(s).

[0108] Dosage forms for transdermal administration of the crystalline compound include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. In embodiments, the crystalline compound is admixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants as may be required.

[0109] In embodiments, ointments, pastes, creams and gels contain, in addition to the crystalline compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof.

[0110] In embodiments, powders and sprays contain, in addition to the crystalline compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. In embodiments, sprays further contain conventional propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0111] In embodiments, the crystalline compounds described herein are formulated as eye drops for ophthalmic administration.

[0112] Alternatively, the compositions and crystalline compounds disclosed herein are administered by aerosol. In embodiments, this is accomplished by preparing an aqueous aerosol, liposomal formulation, or solid particles containing the crystalline compound. In embodiments, a non-aqueous (e.g., fluorocarbon propellant) suspension is used. In embodiments, a sonic nebulizer is used. Typically, aqueous aerosols are made by formulating an aqueous solution or suspension of the crystalline compound with conventional pharmaceutically acceptable carriers and stabilizers. Carriers and stabilizers vary depending on the requirements of the particular subject composition, but typically include non-ionic surfactants (e.g., Tween, Pluronic, or polyethylene glycol), innocuous proteins such as serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars, or sugar alcohols. Aerosols are generally prepared from isotonic solutions.

[0113] Pharmaceutical compositions suitable for parenteral administration comprise the crystalline compound in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders to be reconstituted immediately before use into sterile injectable solutions or dispersions, which in embodiments contain antioxidants, buffers, bacteriostats, solutes or suspending agents or thickening agents that render the formulation isotonic with the blood of the intended recipient.

[0114] Examples of suitable aqueous and non-aqueous carriers used in pharmaceutical compositions include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate and cyclodextrin. Proper fluidity is maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0115] Enteral pharmaceutical formulations comprising a crystalline compound and an enteric material, as well as a pharmaceutically acceptable carrier or excipient thereof, are also contemplated. The enteric material refers to a polymer that is substantially insoluble in the acidic environment of the stomach and is primarily soluble in intestinal fluid at a specific pH. The small intestine is the portion of the digestive tract (intestine) between the stomach and the large intestine, including the duodenum, jejunum, and ileum. The pH of the duodenum is approximately 5.5, the pH of the jejunum is approximately 6.5, and the pH of the distal ileum is approximately 7.5. Thus, the enteric material is not soluble up to a pH of, for example, about 5.0, about 5.2, about 5.4, about 5.6, about 5.8, about 6.0, about 6.2, about 6.4, about 6.6, about 6.8, about 7.0, about 7.2, about 7.4, about 7.6, about 7.8, about 8.0, about 8.2, about 8.4, about 8.6, about 8.8, about 9.0, about 9.2, about 9.4, about 9.6, about 9.8, or about 10.0. Exemplary enteric materials include cellulose acetate phthalate (CAP), hydroxypropyl methylcellulose phthalate (HPMCP), polyvinyl acetate phthalate (PVAP), hydroxypropyl methylcellulose acetate succinate (HPMCAS), cellulose acetate trimellitate, hydroxypropyl methylcellulose succinate, cellulose acetate succinate, cellulose acetate hexahydrophthalate, cellulose propionate phthalate, cellulose acetate maleate, cellulose acetate butyrate, cellulose acetate propionate, copolymers of methyl methacrylic acid and methyl methacrylate, copolymers of methyl acrylate, methyl methacrylate and methacrylic acid, copolymers of methyl vinyl ether and maleic anhydride (Gantrez Examples of suitable enteric dispersions include Eudragit ES series, ethyl methyl acrylate-methyl methacrylate-chlorotrimethylammonium ethyl acrylate copolymers, natural resins such as zein, shellac, and copal colophorium, and several commercially available enteric dispersions (e.g., Eudragit L30D55, Eudragit FS30D, Eudragit L100, Eudragit S100, Kollicoat EMM30D, Estacryl 30D, Coateric, and Aquateric). The solubility of each of the above materials is known or can be readily determined in vitro.

[0116] Doses of compositions containing the crystalline compounds described herein will vary depending on the patient's (e.g., human) condition, i.e., stage of disease, general health, age, and other factors.

[0117] The pharmaceutical composition is administered in a manner appropriate to the disease to be treated (or prevented). The appropriate dose and the appropriate duration and frequency of administration are determined by factors such as the patient's condition, the type and severity of the patient's disease, the particular form of the active ingredient, and the method of administration. In general, an appropriate dose and treatment regimen provides a sufficient amount of the composition to provide therapeutic and / or prophylactic benefit (e.g., improved clinical outcomes such as more frequent complete or partial remissions, or longer disease-free and / or overall survival, or reduced severity of symptoms). Optimal doses are generally determined using experimental models and / or clinical trials. In some embodiments, the optimal dose depends on the patient's body mass, weight, or blood volume.

[0118] Oral dosages typically range from about 1.0 mg to about 1000 mg, one to four or more times per day.

[0119] The crystalline compounds described herein are administered to subjects or patients (animals and humans) in need of such treatment in dosages that provide optimal therapeutic efficacy. It is understood that the dosage required for any particular application will vary from patient to patient, depending not only on the specific composition selected, but also on the route of administration, the nature of the condition being treated, the age and condition of the patient, any concurrent medications or special diets the patient may be following, and other factors; ultimately, the appropriate dosage is left to the discretion of the attending physician. To treat the above-mentioned clinical conditions and diseases, the crystalline compounds disclosed herein are administered orally, subcutaneously, topically, parenterally, by inhalation spray, or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, and vehicles. Parenteral administration includes subcutaneous, intravenous, or intramuscular injection or infusion.

[0120] V. Treatment method In one aspect, a method of treating a subject in need of treatment for a neurodegenerative disorder is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the structure:

[0121] [ka]

[0122] In one aspect, a method of treating a subject in need of treatment for a neurodegenerative disorder is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0123] In one aspect, a method of treating a subject in need of treatment for an inflammatory disease is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the following structure:

[0124] [ka]

[0125] In one aspect, a method of treating a subject in need of treatment for an inflammatory disease is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0126] In embodiments, the inflammatory disease is encephalitis. In embodiments, the inflammatory disease is post-hemorrhagic encephalitis. In embodiments, the inflammatory disease is ocular inflammation. In embodiments, the inflammatory disease is conjunctivitis. In embodiments, the inflammatory disease is allergic conjunctivitis. In embodiments, the inflammatory disease is vernal conjunctivitis. In embodiments, the inflammatory disease is papillary conjunctivitis. In embodiments, the inflammatory disease is Sjogren's syndrome. In embodiments, the inflammatory disease is an inflammatory disease associated with dry eye.

[0127] In one aspect, a method of treating a subject in need of treatment for a demyelinating disease is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the following structure:

[0128] [ka]

[0129] In one aspect, a method of treating a subject in need of treatment for a demyelinating disease is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0130] In embodiments, the demyelinating disease is a demyelinating disease of the central nervous system. In embodiments, the demyelinating disease is multiple sclerosis. In embodiments, the demyelinating disease is a demyelinating disease of the peripheral nervous system.

[0131] In one aspect, a method of treating a subject in need of treatment for a fibrotic disease is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the structure:

[0132] [ka]

[0133] In one aspect, a method of treating a subject in need of treatment for a fibrotic disorder is provided, the method comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound described herein.

[0134] In embodiments, the fibrotic disease is pulmonary fibrosis. In embodiments, the fibrotic disease is dermal fibrosis. In embodiments, the fibrotic disease is hepatic fibrosis. In embodiments, the fibrotic disease is ocular fibrosis. In embodiments, the fibrotic disease is idiopathic pulmonary fibrosis. In embodiments, the fibrotic disease is scleroderma. In embodiments, the fibrotic disease is non-alcoholic steatohepatitis. In embodiments, the fibrotic disease is ocular fibrosis. In embodiments, the fibrotic disease is hypertrophic scars or keloids (e.g., burn-induced or surgical, sarcoidosis, scleroderma, spinal cord injury / fibrosis, myelofibrosis, vascular restenosis, atherosclerosis, arteriosclerosis, Wegener's granulomatosis, mixed connective tissue disease, or Peyronie's disease). In embodiments, the fibrotic disease is iatrogenic pulmonary fibrosis. In embodiments, the fibrotic disease is radiation-induced fibrosis. In embodiments, the fibrotic disease is silicosis-induced pulmonary fibrosis. In embodiments, the fibrotic disease is asbestos-induced pulmonary fibrosis. In embodiments, the fibrotic disease is pleural fibrosis. In embodiments, the fibrotic disease is pulmonary fibrosis associated with SARS-CoV-2 infection and / or COVID-19. In embodiments, the fibrotic disease is pulmonary fibrosis secondary to a systemic inflammatory disease. In embodiments, the fibrotic disease is pulmonary fibrosis secondary to sarcoidosis. In embodiments, the fibrotic disease is intestinal fibrosis. In embodiments, the fibrotic disease is head and neck fibrosis. In embodiments, the fibrotic disease is cirrhosis. In embodiments, the fibrotic disease is alcohol-induced liver fibrosis. In embodiments, the fibrotic disease is endometriosis. In embodiments, the fibrotic disease is spinal fibrosis. In embodiments, the fibrotic disease is myelofibrosis. In embodiments, the fibrotic disease is cardiac fibrosis. In embodiments, the fibrotic disease is perivascular fibrosis. In embodiments, the fibrotic disease is Peyronie's disease. In embodiments, the fibrotic disease is abdominal or intestinal adhesions. In embodiments, the fibrotic disease is bladder fibrosis. In embodiments, the fibrotic disease is nasal fibrosis. In embodiments, the fibrotic disease is fibroblast-mediated fibrosis. In embodiments, the fibrotic disease is renal fibrosis associated with chronic kidney disease (CKD).

[0135] In one aspect, a method of treating a subject in need of treatment for cancer is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the structure:

[0136] [ka]

[0137] In one aspect, a method of treating a subject in need of treatment for cancer is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound described herein.

[0138] In embodiments, the cancer is brain cancer. In embodiments, the cancer is glioblastoma. In embodiments, the cancer is a solid tumor (e.g., of the bladder, intestine, brain, breast, endometrium, heart, kidney, lung, lymphatic tissue (e.g., lymphoma), ovary, pancreas or other endocrine organs (e.g., thyroid), prostate, skin (e.g., melanoma or basal cell carcinoma)), or a hematological tumor (e.g., leukemia), with or without metastasis, at any stage of the disease.In embodiments, the cancer is selected from the group consisting of acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, anal cancer, appendix cancer, astrocytoma, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer (e.g., osteosarcoma or malignant fibrous histiocytoma), brain stem glioma, brain tumor, brain and spinal cord tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, cervical cancer, chronic lymphocytic leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, embryonal tumor, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, Ewing's sarcoma family of tumors, eye cancer, retinoblastoma, gallbladder cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, tumor, GIST), germ cell tumor, glioma, hairy cell leukemia, head and neck cancer, hepatocellular carcinoma, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell tumor (e.g., endocrine pancreatic), Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, liver cancer, lymphoma, medulloblastoma, medulloepithelioma, melanoma, mesothelioma, oral (mouth) cancer, myeloid leukemia, multiple myeloma, nasopharyngeal cancer, Neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma, malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, papillomatosis, parathyroid cancer, penile cancer, pharyngeal cancer, intermediate pineal parenchymal tumor, pineoblastoma, and supratentorial primitive neuroectodermal tumor primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, Sézary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, stomach (e.g., gastric) cancer, supratentorial primitive neuroectodermal tumor, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom's macroglobulinemia, or Wilms' tumor.

[0139] In one aspect, a method of treating a subject in need of treatment for an LPAR1-associated disease is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the following structure:

[0140] [ka]

[0141] In one aspect, a method of treating a subject in need of treatment for an LPAR1-associated disease is provided, the method comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound described herein.

[0142] In embodiments, the LPAR1-associated disease is a neurodegenerative disease. In embodiments, the LPAR1-associated disease is an inflammatory disease. In embodiments, the LPAR1-associated disease is post-hemorrhagic encephalitis. In embodiments, the LPAR1-associated disease is a demyelinating disease. In embodiments, the LPAR1-associated disease is multiple sclerosis. In embodiments, the LPAR1-associated disease is a fibrotic disease. In embodiments, the LPAR1-associated disease is pulmonary fibrosis. In embodiments, the LPAR1-associated disease is idiopathic pulmonary fibrosis. In embodiments, the LPAR1-associated disease is ocular fibrosis. In embodiments, the LPAR1-associated disease is cancer (e.g., brain cancer, ovarian cancer, colon cancer, prostate cancer, breast cancer, melanoma, head and neck cancer, intestinal cancer, colorectal cancer, or thyroid cancer). In embodiments, the LPAR1-associated disease is pain (e.g., neuropathic pain, acute pain, or chronic pain). In embodiments, the LPAR1-associated disease is neuropathic pain. In embodiments, the LPAR1-associated disease is acute pain. In embodiments, the LPAR1-associated disease is chronic pain.

[0143] In embodiments, the LPAR1-associated disease is a respiratory disorder or an allergic disorder. In embodiments, the respiratory disorder or allergic disorder is asthma, peribronchiolar fibrosis, bronchiolitis obliterans, or chronic obstructive pulmonary disease (COPD). In embodiments, the COPD is chronic bronchitis or emphysema, pulmonary hypertension, interstitial pulmonary fibrosis and / or airway inflammation, or cystic fibrosis. In embodiments, the respiratory disease is adult respiratory distress syndrome or allergic (extrinsic) asthma, non-allergic (intrinsic) asthma, acute severe asthma, chronic asthma, clinical asthma, nocturnal asthma, allergen-induced asthma, aspirin-sensitive asthma, exercise-induced asthma, isocapnic hyperventilation, childhood-onset asthma, adult-onset asthma, cough-variant asthma, occupational asthma, steroid-resistant asthma, seasonal asthma, seasonal allergic rhinitis, perennial allergic rhinitis, and hypoxia.

[0144] In embodiments, the LPAR1-associated disease is a nervous system disorder, hi embodiments, the nervous system disorder is Alzheimer's disease, cerebral edema, cerebral ischemia, stroke, multiple sclerosis, neuropathy, Parkinson's disease, neurological conditions seen after blunt or surgical trauma (including postoperative cognitive dysfunction and spinal cord or brainstem injury), degenerative disc disease, or sciatica.

[0145] In embodiments, the LPAR1-related disease is a cardiovascular disorder. In embodiments, the cardiovascular disorder is arrhythmia (e.g., atrial or ventricular), atherosclerosis and its sequelae, angina, cardiac rhythm disorders, myocardial ischemia, myocardial infarction, cardiac or vascular aneurysm, vasculitis, stroke, peripheral occlusive arteriopathy of a limb, organ, or tissue, reperfusion injury after ischemia of the brain, heart, or other organ or tissue, endotoxin, surgical or traumatic shock, hypertension, valvular heart disease, heart failure, abnormal blood pressure, shock, vasoconstriction (including that associated with migraine), vascular abnormalities, or cardiovascular insufficiency limited to a single organ or tissue.

[0146] In embodiments, the LPAR1-associated disease is pulmonary fibrosis, renal fibrosis, liver fibrosis, scarring, asthma, rhinitis, chronic obstructive pulmonary disease (COPD), pulmonary hypertension, interstitial pulmonary fibrosis, arthritis, allergy, psoriasis, inflammatory bowel disease, adult respiratory distress syndrome, myocardial infarction, aneurysm, stroke, cancer, pain, a proliferative disorder, or an inflammatory condition.

[0147] In embodiments, the LPAR1-associated disease is a liver disease. In embodiments, the liver disease is hepatitis C, liver cancer, familial combined hyperlipidemia, non-alcoholic fatty liver disease (NAFLD), progressive familial intrahepatic cholestasis, primary biliary cirrhosis (PBC), or primary sclerosing cholangitis (PSC). In embodiments, the liver disease is primary sclerosing cholangitis (PSC). In embodiments, the liver disease includes portal hypertension. In embodiments, the liver cancer includes hepatocellular carcinoma (HCC), cholangiocarcinoma, angiosarcoma, or hemangiosarcoma. In embodiments, the NAFLD includes steatosis. In embodiments, the NAFLD includes NASH. In an embodiment, the NAFLD or NASH comprises liver fibrosis. In an embodiment, the NAFLD or NASH comprises liver cirrhosis. In an embodiment, the NAFLD or NASH comprises compensated cirrhosis. In an embodiment, the NAFLD or NASH comprises decompensated liver fibrosis. In an embodiment, the NAFLD comprises hepatocellular carcinoma (HCC). In an embodiment, the disease is NASH.

[0148] In one aspect, a method of modulating LPAR1 activity in a subject is provided, the method comprising administering to the subject a crystalline compound of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid having the following structure:

[0149] [ka]

[0150] In one aspect, a method of modulating LPAR1 activity in a subject is provided, the method comprising administering to the subject a crystalline compound described herein.

[0151] VI. Embodiments Embodiment 1. A crystalline compound having the formula:

[0152] [ka] A crystalline compound having characteristic X-ray powder diffraction peaks at about 5.2° 2θ, about 10.4° 2θ, and about 15.6° 2θ.

[0153] Embodiment 2. The crystalline compound of embodiment 1, wherein the crystalline compound has additional characteristic X-ray powder diffraction peaks at about 11.3°2θ, about 12.6°2θ, about 18.4°2θ, about 20.9°2θ, about 22.4°2θ, about 26.1°2θ, about 27.4°2θ, and about 27.6°2θ.

[0154] Embodiment 3. The crystalline compound is selected from the group consisting of about 6.7°2θ, about 7.3°2θ, about 8.6°2θ, about 9.4°2θ, about 11.3°2θ, about 12.0°2θ, about 12.6°2θ, about 13.6°2θ, about 14.9°2θ, about 16.8°2θ, about 17.4°2θ, about 18.4°2θ, about 18.8°2θ, about 19.5°2θ, about 20.1°2θ, about 20.9°2θ, about 21.8°2θ, about 22.4°2θ, about 22.7°2θ, about 23.1°2θ, about 24.1°2θ, about 24.4°2θ, about 24.9°2θ, about 25.4°2θ, about 2Θ, about 26.1°2Θ, about 27.4°2Θ, about 27.6°2Θ, about 28.0°2Θ, about 28.4°2Θ, about 28.7°2Θ, about 29.2°2Θ, about 29.5°2Θ, about 30.0°2Θ, about 30.2°2Θ, about 31.3°2Θ, about 31.8°2Θ, about 32.0°2Θ, about 32.9°2Θ, about 33.9°2Θ, about 34.5°2Θ, about 36.3°2Θ, about 36.9°2Θ, about 37.5°2Θ, about 38.0°2Θ, and about 39.6°2Θ.

[0155] Embodiment 4. The crystalline compound of one of embodiments 1-3, further characterized in that the crystalline compound has a differential scanning calorimetry endotherm onset at about 188.6°C.

[0156] Embodiment 5. A pharmaceutical composition comprising a crystalline compound according to one of embodiments 1 to 4 and a pharmaceutically acceptable excipient.

[0157] Embodiment 6. The pharmaceutical composition of embodiment 5, wherein the pharmaceutical composition does not contain a crystalline form of the compound of formula I other than the crystalline compound described above.

[0158] Embodiment 7. A method of treating a subject in need of treatment for a neurodegenerative disorder, comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound of one of embodiments 1-4.

[0159] Embodiment 8. A method of treating a subject in need of treatment for an inflammatory disease, comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound of one of embodiments 1-4.

[0160] Embodiment 9 The method of embodiment 8, wherein the inflammatory disease is encephalitis.

[0161] Embodiment 10. The method of embodiment 9, wherein the encephalitis is post-hemorrhagic encephalitis.

[0162] Embodiment 11. A method of treating a subject in need of treatment for a demyelinating disease, comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound of one of embodiments 1-4.

[0163] Embodiment 12 The method of embodiment 11, wherein the demyelinating disease is a demyelinating disease of the central nervous system.

[0164] Embodiment 13 The method of embodiment 12, wherein the demyelinating disease is multiple sclerosis.

[0165] Embodiment 14 The method of embodiment 11, wherein the demyelinating disease is a demyelinating disease of the peripheral nervous system.

[0166] Embodiment 15. A method of treating a subject in need of treatment for a fibrotic disease, comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound of one of embodiments 1-4.

[0167] Embodiment 16 The method of embodiment 15, wherein the fibrotic disease is pulmonary fibrosis, skin fibrosis, liver fibrosis, or ocular fibrosis.

[0168] Embodiment 17. The method of embodiment 15, wherein the fibrotic disease is idiopathic pulmonary fibrosis, scleroderma, nonalcoholic steatohepatitis, or ocular fibrosis.

[0169] Embodiment 18. A method of treating a subject in need of treatment for cancer, comprising administering to the subject in need thereof a therapeutically effective amount of a crystalline compound of one of embodiments 1-4.

[0170] Embodiment 19. The method of embodiment 18, wherein the cancer is brain cancer.

[0171] Embodiment 20. The method of embodiment 19, wherein the cancer is glioblastoma.

[0172] Embodiment 21. A method for treating a subject in need of treatment for an LPAR1-associated disorder, comprising administering to the subject in need of treatment a therapeutically effective amount of a crystalline compound described in one of embodiments 1 to 4, wherein the LPAR1-associated disorder is neuropathic pain.

[0173] Embodiment 22. A method of modulating LPAR1 activity in a subject, comprising administering to the subject a crystalline compound of one of embodiments 1-4.

[0174] It is understood that the examples and embodiments described herein are for illustrative purposes only, and that various modifications or changes in light thereof will be suggested to those skilled in the art and are to be included within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes. [Example]

[0175] Polymorph Screening Polymorph screening was performed using the free acid form of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid. The polymorphic behavior of the free acid form was investigated by equilibration, slow cooling, fast cooling, slow evaporation, antisolvent addition, and reverse antisolvent addition experiments.

[0176] Approximately 100 polymorph screening experiments were performed in 15 solvents and their combinations using different crystallization methods, but only one crystalline polymorph, designated Pattern A, was identified.

[0177] Characterization of crystalline compounds Pattern A is the anhydrate. It was obtained from most solvent systems by equilibration, slow cooling, rapid cooling, antisolvent addition, and reverse antisolvent addition experiments. Pattern A is highly crystalline (Figure 1). DSC showed a T of 188.6 °C. 開始 It exhibits a melting peak at 180°C (Figure 2). Decomposition occurs upon melting. TGA shows a weight loss of about 0.4% at about 180°C (Figure 3). 1 H-NMR shows no detectable residual solvent (Figure 4). PLM reveals that the crystalline morphology is plate-like, with particle sizes ranging from 1 to 20 μm (Figure 5). Pattern A is a stable polymorph.

[0178] Compared with the typical XRPD pattern of pattern A and the single crystal data of pattern A (Figure 6), some screening samples obtained from equilibration, rapid cooling, and slow evaporation contained one small additional peak at 10.0°2θ, which may have been caused by some unknown impurities.

[0179] Bulk stability of Pattern A The bulk stability of Pattern A was evaluated over one week at 25° C. / 92% RH in an open container, at 40° C. / 75% RH in an open container, and at 60° C. in a closed container. Pattern A was found to be physically and chemically stable under these conditions over one week ( FIG. 7 ).

[0180] Moisture absorption of pattern A The hygroscopicity of Pattern A was evaluated by dynamic vapor sorption (DVS) testing at 25°C, 40 → 0 → 95 → 0 → 40% RH cycles, with a mass change rate of 0.002% / min. Pattern A was found to be slightly hygroscopic at 80% RH and hygroscopic at 95% RH. Pattern A absorbed approximately 1.2% water at 80% RH and 3.1% water at 95% RH at 25°C (Figure 8). After the DVS test, the resulting sample remained Pattern A (Figure 9).

[0181] Formulation process feasibility for Pattern A The formulation process feasibility of Pattern A was evaluated by compression, milling, and granulation simulation experiments. Pattern A showed good resistance to compression with no morphology change and only a slight decrease in crystallinity (Figure 10). Upon manual milling, Pattern A rapidly lost crystallinity and converted to a substantially amorphous form after 3 minutes of milling with a mortar and pestle (Figure 11). Similarly, upon wet granulation with ethanol or water, the crystallinity of Pattern A significantly decreased, and partial amorphization was observed (Figure 12).

[0182] Polymorph screening experiments Equilibrate at 25°C for 2 weeks Approximately 50 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was equilibrated in 0.4–1.0 mL of solvent at 25 °C for 2 weeks with a stir bar on a magnetic stir plate at a speed of 300–400 rpm.

[0183] The resulting suspension was then filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 2.

[0184] [Table 2]

[0185] Equilibrate at 50°C for 1 week Approximately 50 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was equilibrated in 0.2–1 mL of solvent at 50 °C for 1 week with a stir bar on a magnetic stir plate at a speed of 300–400 rpm.

[0186] The resulting suspension was then filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 3.

[0187] [Table 3]

[0188] Equilibrate at 90°C for 5 days Approximately 50 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was equilibrated in 0.2–0.6 mL of solvent at 90 °C for 5 days with a stir bar on a magnetic stir plate at a speed of 300–400 rpm.

[0189] The resulting suspension was then filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 4.

[0190] [Table 4]

[0191] Equilibration under temperature cycling Approximately 50 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was equilibrated in 0.2 to 1 mL of solvent at 5°C to 50°C for 10 cycles at a heating / cooling rate of 0.1°C / min. Equilibration was performed using a stir bar on a magnetic stir plate at a speed of 400 rpm.

[0192] The resulting suspension was then filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 5.

[0193] [Table 5]

[0194] Crystallization from high-temperature saturated solutions by slow cooling Approximately 30 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was dissolved in a minimum amount of the selected solvent at 50°C. The resulting solution was then filtered through a 0.45 μm syringe membrane filter. The clear solution was cooled to 5°C at 0.1°C / min. The precipitate-free sample at 5°C was further cooled to -20°C.

[0195] The precipitate was collected by centrifugal filtration at 14,000 rpm through a 0.45 μm nylon membrane filter. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 6.

[0196] [Table 6]

[0197] Crystallization from high-temperature saturated solutions by rapid cooling Approximately 30 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was dissolved in a minimum amount of the selected solvent at 50°C. The resulting solution was then filtered through a 0.45 μm syringe membrane filter. The clear solution was placed in an ice bath at 0°C and stirred. The precipitate-free sample at 0°C was further cooled to -20°C.

[0198] The precipitate was collected by centrifugal filtration at 14,000 rpm through a 0.45 μm nylon membrane filter. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 7.

[0199] [Table 7]

[0200] Crystallization by slow addition of antisolvent to homogeneous solution Approximately 30 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was dissolved in a minimum amount of selected solvent at ambient temperature (approximately 25 °C). The resulting solution was then filtered through a 0.45 μm syringe membrane filter. Two to four times the volume of anti-solvent was slowly added to the clear solution. The precipitate-free sample at 25 °C was further cooled to -20 °C.

[0201] The precipitate was collected by centrifugal filtration at 14,000 rpm through a 0.45 μm nylon membrane filter. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 8.

[0202] [Table 8]

[0203] Crystallization by rapid addition of a homogeneous solution to a poor solvent Approximately 30 mg of trans-4-3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was dissolved in a minimum amount of selected solvent at ambient temperature (approximately 25°C). The resulting solution was then filtered through a 0.45 μm syringe membrane filter. The clear solution was quickly added to 4 volumes of anti-solvent. The precipitate-free sample at 25°C was further cooled to -20°C.

[0204] The precipitate was collected by centrifugal filtration at 14,000 rpm through a 0.45 μm nylon membrane filter. The solid portion (wet cake) was examined by XRPD. The results are summarized in Table 9.

[0205] [Table 9]

[0206] Crystallization by slow evaporation at room temperature Approximately 30 mg of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid was dissolved in 0.2-5.0 mL of solvent. The resulting solution was then filtered through a 0.45 μm syringe membrane filter. The clear solution was allowed to slowly evaporate under ambient conditions (approximately 10-25 °C, 20-30% RH).

[0207] The solid residue was examined by XRPD and the results are summarized in Table 10.

[0208] [Table 10]

[0209] Characterization of crystalline compounds experiments Bulk stability determination experiments The Pattern A polymorph was placed in an open container at 25°C / 92% RH, in an open container at 40°C / 75% RH, and in a closed container at 60°C for one week. The stressed samples were characterized by XRPD and HPLC and examined for color change. The results are summarized in Table 11.

[0210] [Table 11]

[0211] Water adsorption and desorption experiments The water sorption and desorption behavior of the Pattern A polymorph was investigated by DVS at 25°C using a cycle of 40-0-95-0-40% RH, dm / dt 0.002, a minimum equilibration time of 60 minutes, and a maximum equilibration time of 360 minutes. XRPD was measured after the DVS test to determine the morphology change. The results are summarized in Table 12.

[0212] [Table 12]

[0213] Compression simulation experiment Approximately 10 mg of the Pattern A polymorph was compressed under 5 MPa and 10 MPa for 5 minutes using a hydraulic press. Potential morphological changes and crystallinity were assessed by XRPD. The results are summarized in Table 13.

[0214] [Table 13]

[0215] Dry grinding simulation experiment Approximately 20 mg of the Pattern A polymorph was manually ground with a mortar and pestle for 1 minute, 3 minutes, and 5 minutes. Potential morphological changes and crystallinity were assessed by XRPD. The results are summarized in Table 14.

[0216] [Table 14]

[0217] Wet granulation simulation experiment Water or ethanol was added dropwise to approximately 20 mg of the Pattern A polymorph until the sample was sufficiently wet. The wet sample was gently crushed with a mortar and pestle. The granulated sample was allowed to dry at ambient conditions for 3 minutes. Potential morphological changes and crystallinity were assessed by XRPD. The results are summarized in Table 15.

[0218] [Table 15]

[0219] Preparation of polymorphs Preparation of Pattern A 1.0 g of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid (see below) was weighed into a 40 mL glass vial. 10 mL of ACN was added to the vial with stirring at 50° C. for approximately 1 minute to obtain a suspension.

[0220] After stirring at 50°C for 4 days, the solid was collected through a 0.45 μm nylon membrane filter by centrifugation at 4,000 rpm, and then dried under vacuum at 25°C for approximately 20 hours.

[0221] 748 mg of Pattern A was obtained as an off-white solid in 76% yield.

[0222] Preparation of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid

[0223] [ka]

[0224] Step 1: In a dry round-bottom flask equipped with a magnetic stirrer, 6-methoxy-3-nitropyridin-2-ol (1 equiv., Combi-Blocks) was suspended in acetonitrile (0.10 M). Sodium hydride (60% w / w dispersion in paraffin oil, 2.8 equiv., Sigma-Aldrich) was then added rapidly in one portion, and the resulting mixture was stirred at room temperature for 10 minutes to give a brownish-yellow suspension. 2,2-Difluoro-2-(fluorosulfonyl)acetic acid (1.8 equiv., Sigma-Aldrich) was then added dropwise over 5 minutes, during which a mild exotherm was observed. After stirring for 16 hours, another aliquot of 2,2-difluoro-2-(fluorosulfonyl)acetic acid (1.8 equiv., Sigma-Aldrich) was added dropwise over 5 minutes. After stirring at room temperature for an additional 48 hours, the crude reaction mixture was carefully quenched with water and then diluted with a 1:1 (v / v) solution of ethyl acetate and hexane. The organic layer was then separated, washed successively with saturated aqueous NaHCO, water, and brine, dried over MgSO, filtered, and the filtrate was concentrated in vacuo. The crude product thus obtained was purified by column chromatography (SiO, gradient elution: Hex → 1:1 (v / v) Hex:EtOAc) to give 2-(difluoromethoxy)-6-methoxy-3-nitropyridine as a yellow solid (75% yield).

[0225] Step 2: In a dry round-bottom flask equipped with a magnetic stirrer, 2-(difluoromethoxy)-6-methoxy-3-nitropyridine (1 equiv.) from the previous step was dissolved in methanol (0.17 M). The resulting yellow solution was then deoxygenated by subsurface purging with nitrogen for 10 minutes, after which palladium (10% w / w on activated carbon, dry, 0.08 equiv., Sigma-Aldrich) was added in one portion. The resulting black suspension was then subsurface purged with hydrogen for 10 minutes, after which it was stirred at room temperature under a static hydrogen atmosphere (maintained by a balloon) for 90 minutes. The reaction was then diluted with EtOAc and filtered through a bed of Celite wetted with dichloromethane. The insoluble material was further washed with EtOAc. The filtrate thus obtained was concentrated in vacuo to afford 2-(difluoromethoxy)-6-methoxypyridin-3-amine as a reddish-brown solid (>99% yield).

[0226] Step 3: In a dry round-bottom flask equipped with a magnetic stirrer, 2-(difluoromethoxy)-6-methoxypyridin-3-amine (1 equivalent) from the previous step and pyridine (3 equivalents, Sigma-Aldrich) were combined in anhydrous dichloromethane (0.086 M). To this was then added phosgene (15% w / w solution in toluene, 1.5 equivalents, Sigma-Aldrich) dropwise at room temperature, and the resulting solution was stirred at room temperature for 15 minutes. The volatiles were then removed in vacuo, and the crude (2-(difluoromethoxy)-6-methoxypyridin-3-yl)carbamic acid chloride thus obtained was redissolved in anhydrous dichloromethane (0.12 M). This solution was then added dropwise to another dichloromethane suspension (0.12 M) of trans-ethyl 4-((2-isopropylphenyl)amino)cyclohexane-1-carboxylate (1 equivalent, intermediate amine 1, see below), pyridine (3 equivalents, Sigma-Aldrich), and freshly activated 4 Å molecular sieves at room temperature. The resulting mixture was stirred at room temperature for 24 h, after which the reaction was quenched with water. The aqueous layer was separated and back-extracted with EtOAc. The combined organic extracts were dried over MgSO, filtered, and the filtrate was concentrated in vacuo. The crude product thus obtained was purified by column chromatography (SiO, gradient elution: Hex → 7:3 (v / v) Hex:EtOAc) to afford trans-ethyl 4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylate as a white foam (81% yield).

[0227] Step 4: In a round-bottom flask equipped with a magnetic stirrer, trans-ethyl 4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylate (1 equivalent) from the previous step was dissolved in a 1:1 (v / v) solution of THF and methanol (0.11 M). To this colorless solution was then added lithium hydroxide (1 M aqueous solution, 5 equivalents), and the resulting mixture was heated at 45° C. for 3 h. Upon cooling to room temperature, the reaction mixture was carefully quenched with HCl (1 M aqueous solution, 5 equivalents), and the volatiles were removed in vacuo. The resulting suspension was then back-extracted with EtOAc. The combined organic extracts were washed successively with water and brine, dried over MgSO4, and filtered. The filtrate was concentrated in vacuo to give a white solid, which was then vigorously stirred in hot isopropanol until homogeneous. The resulting solution was then cooled until it became cloudy, at which point an equal volume of heptane was added with vigorous stirring. After stirring at room temperature for 16 hours, the suspension was filtered to give the title compound as a white solid. LCMS: m / z=478.1 [M+H] + ; 1 H NMR (DMSO-d6): δ=8.08(d,J=8.7Hz,1H),7.56~7.49(m,2H),7.36(t,J=72.9Hz, 1H),7.35(td,J=7.2,1.8Hz,1H),7.25(dd,J=8.1,1.2Hz,1H),6.54(d,J=8.7Hz ,1H),6.29(s,1H),4.30~4.21(m,1H),3.82(s,3H),3.22(sept,J=6.9Hz,1H),2 .24~1.85(m,5H),1.62~1.46(m,3H),1.29(d,J=6.9Hz,3H),1.22~1.05(m,4H).

[0228] Preparation of intermediate amine 1: trans-ethyl 4-((2-isopropylphenyl)amino)cyclohexane-1-carboxylate

[0229] [ka] In a thick-walled glass reaction vessel equipped with a magnetic stirrer and a Teflon screw cap, trans-ethyl 4-aminocyclohexanecarboxylate hydrochloride (1 equiv., ChemScene), 1-iodo-2-isopropylbenzene (1.4 equiv., Combi-Blocks), tris(dibenzylideneacetone)dipalladium(0) (0.1 equiv., Sigma-Aldrich), 2-cyclohexylphosphino-2',6'-bis(N,N-dimethylamino)biphenyl (0.2 equiv., Combi-Blocks), and cesium carbonate (4 equiv., Sigma-Aldrich) were combined in 1,4-dioxane (0.12 M). The resulting purple suspension was then deoxygenated by subsurface purging with nitrogen for 10 minutes, after which the reaction vessel was tightly sealed and heated at 90 °C for 48 h. The resulting orange-brown suspension was cooled to room temperature, diluted with tert-butyl methyl ether, and washed successively with water and brine. The organic extract thus obtained was then dried over MgSO4, treated with charcoal, filtered through a bed of Celite, and the filtrate was concentrated in vacuo. The crude product thus obtained was purified by column chromatography (SiO2, gradient elution: Hex → 4:1 (v / v) Hex:EtOAc) to afford the title compound as a golden yellow oil (67% yield).

[0230] Biological evaluation: In vitro functional assay of lysophosphatidic acid receptor 1 activity Primary compound plates were prepared in 100% DMSO (Sigma-Aldrich), secondary compound plates were prepared at 10x concentrations in DMEM (Invitrogen), and tertiary compound plates were prepared in HBSS (Ca +2 / Mg +2The cells were prepared at a 3x concentration in assay buffer containing 0.1% BSA (Sigma-Aldrich) and 0.1% FBS (Invitrogen). Fluo-4 NW calcium assay dye (Invitrogen) was prepared in assay buffer according to the manufacturer's recommendations. B103 cells (J. Chun lab, UCSD) stably expressing human LPAR1 were grown to confluence in DMEM medium (Invitrogen) containing 10% FBS (ATCC), 10% penicillin-streptomycin (Sigma-Aldrich), and 50 μg Geneticin (Sigma-Aldrich) and detached with Accutase (Sigma-Aldrich) prior to the assay. Freshly detached cells were resuspended in growth medium and plated at 5 × 10 cells per well into black, clear-bottom 96-well plates (Costar) containing compound. 4 Cells were seeded at a density of 1000 cells / well. After seeding, the cells were left at room temperature for 30 minutes and then transferred to a 37°C, 5% CO2 incubator for 24 hours. After 24 hours, the growth medium was removed and freshly prepared Fluo-4 NW calcium assay dye was added to the cells. Compounds (i.e., the tertiary compound plate) were then added back to the dye / cells and returned to the incubator for 30 minutes, then maintained at room temperature for another 30 minutes. Finally, EC 80 Lysophosphatidic acid (18:1) was added at a concentration of 0.05 mg / mL, and calcium flux was measured using a FlexStation 3 (Molecular Devices). Sigmoidal dose-response curves were generated by measuring luminescence over 45 seconds and calculating the area under the curve. Dose-response curves and IC 50 Values ​​were generated using Prism (GraphPad). Compounds were tested at final concentrations ranging from 100 pM to 10 μM in 0.1% DMSO.

[0231] Lysophosphatidic acid receptor 1 IC of trans-4-(3-(2-(difluoromethoxy)-6-methoxypyridin-3-yl)-1-(2-isopropylphenyl)ureido)cyclohexane-1-carboxylic acid 50 was found to be 10 nM.

[0232] Although the foregoing has been described in some detail by way of illustration and example, for purposes of clarity and understanding, it will be understood by those skilled in the art that numerous and various modifications may be made without departing from the spirit of the present disclosure. It is therefore to be clearly understood that the forms disclosed herein are illustrative only and are not intended to limit the scope of the present disclosure, but rather to embrace all modifications and alternatives which fall within the true scope and spirit of the invention.

Claims

1. A crystalline compound having the formula: 【Chemistry 1】 A crystalline compound having characteristic X-ray powder diffraction peaks at about 5.2° 2θ, about 10.4° 2θ, and about 15.6° 2θ.

2. 2. The crystalline compound of claim 1, wherein the crystalline compound has additional characteristic X-ray powder diffraction peaks at about 11.3°2θ, about 12.6°2θ, about 18.4°2θ, about 20.9°2θ, about 22.4°2θ, about 26.1°2θ, about 27.4°2θ, and about 27.6°2θ.

3. The crystalline compound is selected from the group consisting of about 6.7°2θ, about 7.3°2θ, about 8.6°2θ, about 9.4°2θ, about 11.3°2θ, about 12.0°2θ, about 12.6°2θ, about 13.6°2θ, about 14.9°2θ, about 16.8°2θ, about 17.4°2θ, about 18.4°2θ, about 18.8°2θ, about 19.5°2θ, about 20.1°2θ, about 20.9°2θ, about 21.8°2θ, about 22.4°2θ, about 22.7°2θ, about 23.1°2θ, about 24.1°2θ, about 24.4°2θ, about 24.9°2θ, about 25.4°2θ, about 26. 2θ, about 6.1°2θ, about 27.4°2θ, about 27.6°2θ, about 28.0°2θ, about 28.4°2θ, about 28.7°2θ, about 29.2°2θ, about 29.5°2θ, about 30.0°2θ, about 30.2°2θ, about 31.3°2θ, about 31.8°2θ, about 32.0°2θ, about 32.9°2θ, about 33.9°2θ, about 34.5°2θ, about 36.3°2θ, about 36.9°2θ, about 37.5°2θ, about 38.0°2θ, and about 39.6°2θ.

4. 10. The crystalline compound of claim 1, wherein the crystalline compound is further characterized by a differential scanning calorimetry endotherm onset at about 188.6°C.

5. A pharmaceutical composition comprising the crystalline compound of any one of claims 1 to 4 and a pharmaceutically acceptable excipient.

6. 6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition does not contain any crystalline form of the compound of Formula I other than the crystalline compound.

7. 10. A method of treating a subject in need of treatment for a neurodegenerative disorder, comprising administering to said subject in need thereof a therapeutically effective amount of the crystalline compound of any one of claims 1 to 4.

8. 10. A method of treating a subject in need of treatment for an inflammatory disease, comprising administering to said subject in need thereof a therapeutically effective amount of the crystalline compound of any one of claims 1 to 4.

9. 9. The method of claim 8, wherein the inflammatory disease is encephalitis.

10. 10. The method of claim 9, wherein the encephalitis is post-hemorrhagic encephalitis.

11. 10. A method of treating a subject in need of treatment for a demyelinating disease, comprising administering to said subject in need thereof a therapeutically effective amount of the crystalline compound of any one of claims 1 to 4.

12. 12. The method of claim 11, wherein the demyelinating disease is a demyelinating disease of the central nervous system.

13. 13. The method of claim 12, wherein the demyelinating disease is multiple sclerosis.

14. 12. The method of claim 11, wherein the demyelinating disease is a demyelinating disease of the peripheral nervous system.

15. 10. A method of treating a subject in need of treatment for a fibrotic disease, comprising administering to said subject in need thereof a therapeutically effective amount of the crystalline compound of any one of claims 1 to 4.

16. 16. The method of claim 15, wherein the fibrotic disease is pulmonary fibrosis, skin fibrosis, liver fibrosis, or ocular fibrosis.

17. 16. The method of claim 15, wherein the fibrotic disease is idiopathic pulmonary fibrosis, scleroderma, nonalcoholic steatohepatitis, or ocular fibrosis.

18. 10. A method of treating a subject in need of treatment for cancer, comprising administering to said subject in need thereof a therapeutically effective amount of the crystalline compound of any one of claims 1 to 4.

19. 19. The method of claim 18, wherein the cancer is brain cancer.

20. 20. The method of claim 19, wherein the cancer is glioblastoma.

21. A method for treating a subject in need of treatment for an LPAR1-associated disease, comprising administering a therapeutically effective amount of the crystalline compound of any one of claims 1 to 4 to the subject in need of treatment, wherein the LPAR1-associated disease is neuropathic pain.

22. A method of modulating LPAR1 activity in a subject, comprising administering to said subject a crystalline compound of any one of claims 1 to 4.

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