Crystalline forms of glucosylceramide synthase inhibitors and uses thereof

Abstract

The present disclosure relates to a crystalline form of (E)-N-((1R, 2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl) prop-2-yl)-2-(6-chloronaphthalene-2-yl)-2-(hydroxyimino) acetamide, a process for preparing the crystalline form, a pharmaceutical composition comprising the crystalline form, and uses thereof. The present disclosure relates to a crystalline form of (E)-N-((1R, 2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl) prop-2-yl)-2-(6-chloronaphthalene-2-yl)-2-(hydroxyimino) acetamide.

Description

Technical Field

[0001] The present disclosure generally relates to crystal forms of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, methods for preparing such crystal forms, pharmaceutical compositions comprising such crystal forms, and uses thereof. Background Art

[0002] Glucosylceramide synthase (GCS) is a key enzyme that catalyzes the initial glycosylation step in the biosynthesis of glucosylceramide-based glycosphingolipids (GSLs), namely, the transfer of glucose from UDP-glucose (UDP-Glc) to ceramide to form glucosylceramide. GCS inhibitors have been proposed for the treatment of various diseases (see, e.g., WO 2005068426).

[0003] As described in PCT Publication No. WO2015042397A1, (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide (hereinafter referred to as "Compound 1") is a GCS inhibitor, which can be used for preventing or treating GCS-related diseases, disorders or conditions. PCT Publication No. WO2015042397A1 is incorporated herein by reference in its entirety. The structure of Compound 1 is shown below:

[0004]

[0005] A compound can exist in one or more crystal forms. The crystal forms of a drug substance can have different physical properties, including melting point, solubility, dissolution rate, optical and mechanical properties, vapor pressure, hygroscopicity, particle shape, density, and flowability, etc. These properties have a direct impact on the ability to process and / or manufacture the compound into a drug product. Crystal forms can also exhibit different stabilities and bioavailabilities. During drug development, the most stable crystal form of a drug product is usually selected based on the minimum possibility of conversion into another crystal form and its greater chemical stability. To ensure the quality, safety, and efficacy of a drug product, it is important to select a crystal form that is stable, reproducibly manufacturable, and has good physicochemical properties.

[0006] Therefore, there is still a need for a stable form of Compound 1 with favorable chemical and physical properties. Summary of the Invention

[0007] On the one hand, the present disclosure provides various crystal forms of Compound 1 and its anhydrates, hydrates, solvates, and salts.

[0008] In some embodiments, the present disclosure provides a crystalline form A of Compound 1, wherein the crystalline form A is characterized in that its X-ray powder diffraction (XRPD) pattern includes one or more peaks selected from the group consisting of about 18.883°, about 10.889°, and about 21.309° 2θ.

[0009] In some embodiments, the present disclosure provides a crystalline form B of Compound 1, wherein the crystalline form B is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 6.244°, about 18.679°, and about 12.445° 2θ.

[0010] In some embodiments, the present disclosure provides a crystalline form C of Compound 1, wherein the crystalline form C is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 6.656°, about 19.994°, and about 26.732° 2θ.

[0011] In some embodiments, the present disclosure provides a crystalline form D of Compound 1, wherein the crystalline form D is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 16.956°, about 5.883°, and about 22.181° 2θ.

[0012] In some embodiments, the present disclosure provides a crystalline form E of Compound 1, wherein the crystalline form E is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 18.089°, about 26.315°, and about 19.775° 2θ.

[0013] In some embodiments, the present disclosure provides a crystalline form A of the L-malate salt of Compound 1, wherein the crystalline form A of the L-malate salt is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 19.349°, about 22.304°, and about 20.078° 2θ.

[0014] In some embodiments, the present disclosure provides a crystalline form A of the L-tartrate salt of Compound 1, wherein the crystalline form A of the L-tartrate salt is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 18.564°, about 21.685°, and about 18.774° 2θ.

[0015] On the other hand, the present disclosure provides a pharmaceutical composition comprising the crystalline form of Compound 1 provided herein.

[0016] On the other hand, the present disclosure provides a method for preparing the crystalline form of Compound 1 or the pharmaceutical composition provided herein.

[0017] On the other hand, the present disclosure provides the use of a crystalline form of compound 1 provided herein or a pharmaceutical composition in the preparation of a medicament for (a) preventing or treating a disease, disorder or condition associated with abnormal activity / level of the enzyme GCS; (b) preventing or treating a GCS-mediated disease, disorder or condition; (c) inhibiting the activity / level of the enzyme GCS; or (d) non-therapeutically inhibiting the activity / level of the GCS enzyme in vitro.

[0018] On the other hand, the present disclosure provides a method for inhibiting GCS activity, the method comprising administering to a subject in need thereof an effective amount of a crystalline form of compound 1 provided herein or a pharmaceutical composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Shows the XRPD pattern of crystalline form A of compound 1.

[0020] Figure 2 Shows the TGA curve of crystalline form A of compound 1.

[0021] Figure 3 Shows the DSC curve of crystalline form A of compound 1.

[0022] Figure 4 Shows the 1 1H-NMR spectrum of crystalline form A of compound 1.

[0023] Figure 5 Shows the XRPD pattern of crystalline form B of compound 1.

[0024] Figure 6 Shows the TGA curve of crystalline form B of compound 1.

[0025] Figure 7 Shows the DSC curve of crystalline form B of compound 1.

[0026] Figure 8 Shows the 1 1H-NMR spectrum of crystalline form B of compound 1.

[0027] Figure 9 Shows the XRPD pattern of crystalline form C of compound 1.

[0028] Figure 10 Shows the TGA curve of crystalline form C of compound 1.

[0029] Figure 11 Shows the DSC curve of crystalline form C of compound 1.

[0030] Figure 12 Shows the 1 1H-NMR spectrum of crystalline form C of compound 1.

[0031] Figure 13 The XRPD pattern of crystalline form D of Compound 1 is shown.

[0032] Figure 14 The TGA curve of crystalline form D of Compound 1 is shown.

[0033] Figure 15 The DSC curve of crystalline form D of Compound 1 is shown.

[0034] Figure 16 The 1 H-NMR spectrum of crystalline form D of Compound 1 is shown.

[0035] Figure 17 The XRPD pattern of crystalline form E of Compound 1 is shown.

[0036] Figure 18 The XRPD pattern of crystalline form A of L-malate of Compound 1 is shown.

[0037] Figure 19 The TGA curve of crystalline form A of L-malate of Compound 1 is shown.

[0038] Figure 20 The DSC curve of crystalline form A of L-malate of Compound 1 is shown.

[0039] Figure 21 The 1 H-NMR spectrum of crystalline form A of L-malate of Compound 1 is shown.

[0040] Figure 22 The XRPD pattern of crystalline form A of L-tartrate of Compound 1 is shown.

[0041] Figure 23 The TGA curve of crystalline form A of L-tartrate of Compound 1 is shown.

[0042] Figure 24 The DSC curve of crystalline form A of L-tartrate of Compound 1 is shown.

[0043] Figure 25 The 1 H-NMR spectrum of crystalline form A of L-tartrate of Compound 1 is shown.

[0044] Figure 26 The DVS isotherm diagram of crystalline form A of Compound 1 at 25 °C is shown.

[0045] Figure 27 The XRPD overlay diagram of crystalline form A of Compound 1 before and after DVS is shown.

[0046] Figure 28The DVS isotherm diagram of Form A of the L-malate salt of Compound 1 at 25 °C is shown.

[0047] Figure 29 The XRPD overlay diagrams of Form A of the L-malate salt of Compound 1 before and after DVS are shown.

[0048] Figure 30 The DVS isotherm diagram of Form A of the L-tartrate salt of Compound 1 at 25 °C is shown.

[0049] Figure 31 The XRPD overlay diagrams of Form A of the L-tartrate salt of Compound 1 before and after DVS are shown.

[0050] Figure 32 A - C show the results of the efficacy tests of the GCS inhibitor Eliglustat ( Figure 32 A), Venglustat ( Figure 32 B), and Form A of Compound 1 ( Figure 32 C) using MDCK cell lysates.

[0051] Figure 33 A - D show the results of the efficacy tests of the GCS inhibitor Eliglustat ( Figure 33 A), Venglustat ( Figure 33 B), Form A of Compound 1 ( Figure 33 C), and the geometric isomer of Compound 1 ( Figure 33 D) using live K - 562 cells.

[0052] Figure 34 The results of the GCS activity assay of L6 cell lysates are shown. Detailed Description

[0053] Throughout this disclosure, the articles "a / an" and "the" are used herein to refer to one or more than one (i.e., at least one) grammatical object of the article. For example, "crystal form" refers to one crystal form or more than one crystal form.

[0054] The following description of the disclosure is only intended to illustrate the various embodiments of the disclosure. Thus, the specific modifications discussed should not be construed as limiting the scope of the disclosure. It will be apparent to those skilled in the art that various equivalents, changes, and modifications can be made without departing from the scope of the disclosure, and it should be understood that such equivalent embodiments will be included herein. All documents cited herein, including published publications, patents, and patent applications, are incorporated herein by reference in their entirety.

[0055] I.Definition

[0056] Unless otherwise indicated, as used in this specification and the following claims, the words "comprise / comprising" and "include / including / includes" are intended to specify the presence of the stated feature, integer, component, or step, but these words do not preclude the presence or addition of one or more other features, integers, components, steps, or groups.

[0057] As used herein, the term "effective amount" or "therapeutically effective amount" means an amount of an agent or compound administered that will, to some extent, alleviate one or more of the symptoms of the disease or disorder being treated. The results include alleviation and / or amelioration of the signs, symptoms, or causes of the disease and / or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound disclosed herein that is required to effect a clinically significant reduction in the symptoms of a disease. Any suitable technique (such as dose escalation studies) is used to determine the appropriate "effective" amount in any individual case.

[0058] As used herein, the term "excipient" or "pharmaceutically acceptable excipient" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating material. In one embodiment, each component is "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of the pharmaceutical formulation and suitable for contact with the tissues or organs of humans and animals without undue toxicity, irritation, allergic response, immunogenicity, or other problems or complications commensurate with a reasonable benefit / risk ratio. See, e.g., Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 6th Edition, edited by Rowe et al., The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd Edition, edited by Ash and Ash, Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd Edition, edited by Gibson, CRC Press LLC: Boca Raton, FL, 2009.

[0059] As used herein, the term "pharmaceutically acceptable salt" refers to a preparation of a compound that does not cause significant irritation to the organism to which it is administered and does not eliminate the biological activity and properties of the compound. In some cases, pharmaceutically acceptable salts are obtained by reacting the compounds described herein with acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc. In some cases, pharmaceutically acceptable salts are obtained by reacting a compound having an acidic group described herein with a base to form a salt (such as an ammonium salt), an alkali metal salt (such as a sodium salt or a potassium salt), an alkaline earth metal salt (such as a calcium salt or a magnesium salt), a salt of an organic base (such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine), and a salt with an amino acid such as arginine, lysine, or other previously determined methods. Pharmaceutically acceptable salts are not particularly limited as long as they can be used in drugs. Examples of salts formed by the compounds of the present invention with bases include the following: salts formed with inorganic bases such as sodium, potassium, magnesium, calcium, and aluminum; salts formed with organic bases such as methylamine, ethylamine, and ethanolamine; salts formed with basic amino acids such as lysine and ornithine; and ammonium salts. The salts may be acid addition salts, which are specifically exemplified by acid addition salts with the following: inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, and ethanesulfonic acid; acidic amino acids such as aspartic acid and glutamic acid.

[0060] The term "pharmaceutical composition" refers to a mixture of a compound described herein with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. Pharmaceutical compositions facilitate the administration of the compound to an organism. A variety of techniques for administering compounds that exist in the art include, but are not limited to, intravenous, oral, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0061] The term "subject" refers to an animal, including but not limited to primates (e.g., humans), monkeys, cattle, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, or mice. The terms "subject" and "patient" are used interchangeably herein, in the context of, for example, mammalian subjects such as humans.

[0062] In the context of treating a disease or disorder, the terms "treat", "treating", and "treatment" mean including alleviating or eliminating the disorder, disease, or condition, or one or more symptoms associated with the disorder, disease, or condition; or slowing the progression, spread, or worsening of the disease, disorder, or condition or one or more of its symptoms. Generally, the beneficial effects obtained by a subject from a therapeutic agent do not completely cure the disease, disorder, or condition.

[0063] Regarding a specific crystalline form of Compound 1, in certain embodiments, as used herein, the term "substantially pure" means that the specific crystalline form of Compound 1 contains less than 99 wt%, less than 95 wt%, less than 90 wt%, less than 85 wt%, less than 80 wt%, less than 75 wt%, less than 70 wt%, less than 65 wt%, less than 60 wt%, less than 55 wt%, less than 50 wt%, less than 45 wt%, less than 40 wt%, less than 35 wt%, less than 30 wt%, less than 25 wt%, less than 20 wt%, less than 15 wt%, less than 10 wt%, less than 9 wt%, less than 8 wt%, less than 7 wt%, less than 6 wt%, less than 5 wt%, less than 4 wt%, less than 3 wt%, less than 2 wt%, less than 1 wt% of other substances, including other crystalline forms and / or impurities. For example, impurities can include by-products, reaction starting materials, reagents from chemical reactions, contaminants, degradation products, water, or solvents, etc. In some embodiments, if the specific crystalline form of Compound 1 is substantially pure, this means that the crystalline form of Compound 1 contains less than 10 wt%, less than 9 wt%, less than 8 wt%, less than 7 wt%, less than 6 wt%, less than 5 wt%, less than 4 wt%, less than 3 wt%, less than 2 wt%, less than 1 wt% of other substances.

[0064] As used herein, the term "polymorph" refers to different crystal structures (solvated or non-solvated forms) in which a compound can crystallize. For example, Compound 1 of the present disclosure can crystallize to form different crystal structures, i.e., polymorphs.

[0065] As used herein, the term "X-ray powder diffraction pattern" or "XRPD pattern" refers to an x-y plot where the x-axis is the diffraction angle (i.e., °2θ) and the y-axis is the intensity. The peaks in this pattern can be used to characterize crystalline solid forms. As with any data measurement, there is variability in XRPD data. The data is typically represented only by the diffraction angle of the peaks and does not include the peak intensity, because the peak intensity can be particularly sensitive to sample preparation (e.g., particle size, moisture content, solvent content, and preferred orientation effects affect sensitivity), and thus samples of the same material prepared under different conditions may produce slightly different patterns; this variability is generally greater than the variability in the diffraction angle. The variability in the diffraction angle can also be sensitive to sample preparation. Other sources of variability come from instrument parameters and the processing of the raw X-ray data: different X-ray instruments operate using different parameters, and these parameters can result in slightly different XRPD patterns for the same solid form, and similarly different software packages process the X-ray data differently, which also results in variability. These and other sources of variability are known to those of ordinary skill in the pharmaceutical art. Due to such sources of variability, a variability of ±0.3° 2θ in the diffraction angle in the XRPD pattern is typically allowed. For example, in the present disclosure, when referring to the diffraction angle °2θ, the term "about" means that a variability of ±0.3° 2θ can be assigned to the value mentioned, e.g., the variability can be -0.3°, -0.29°, -0.28°, -0.27°, -0.26°, -0.25°, -0.24°, -0.23°, -0.22°, -0.21°, -0.2°, -0.19°, -0.18°, -0.17°, -0.16°, -0.15°, -0.14°, -0.13°, -0.12°, -0.11°, -0.1°, -0.09°, -0.08°, -0.07°, -0.06°, -0.05°, -0.04°, -0.03°, -0.02°, -0.01°, 0.00°, 0.01°, 0.02°, 0.03°, 0.04°, 0.05°, 0.06°, 0.07°, 0.08°, 0.09°, 0.1°, 0.11°, 0.12°, 0.13°, 0.14°, 0.15°, 0.16°, 0.17°, 0.18°, 0.19°, 0.2°, 0.21°, 0.22°, 0.23°, 0.24°, 0.25°, 0.26°, 0.27°, 0.28°, 0.29° or 0.3°, preferably ±0.2°.

[0066] As used herein, the term "differential scanning calorimetry" or "DSC" refers to measuring the temperature difference and heat flow difference between a sample and a reference material during heating or isothermal holding of the sample to characterize all physical and chemical changes related to thermal effects, thereby obtaining phase transition information of the sample.

[0067] In the present disclosure, when referring to spectra (e.g., XRPD spectra, DSC curves, TGA curves, HPLC spectra, 1 1H NMR spectra, DVS spectra, etc.), the term "substantially similar to" or "substantially as shown" means that the basic characteristic information or main characteristic information (e.g., peak position, intensity, etc.) of the spectrum is consistent with the information described in the spectrum, and it is not required that all features of the spectrum be exactly the same as the features described in the spectrum.

[0068] II. Polymorph A of Compound 1

[0069] On the one hand, the present disclosure provides polymorph A of compound 1. In some embodiments, the present disclosure provides substantially pure polymorph A of compound 1. In some embodiments, compound 1 is an unsolvated polymorph, e.g., polymorph A of compound 1 is an anhydrate.

[0070] In some embodiments, the XRPD spectrum of polymorph A of compound 1 includes one or more (e.g., two, three) peaks selected from the group consisting of: at about 18.883°, about 10.889°, and about 21.309° 2θ. For example, the XRPD spectrum of polymorph A of compound 1 includes a peak at about 18.883° 2θ. Again, for example, the XRPD spectrum of polymorph A of compound 1 includes a peak at about 10.889° 2θ. Again, for example, the XRPD spectrum of polymorph A of compound 1 includes a peak at about 21.309° 2θ. Again, for example, the XRPD spectrum of polymorph A of compound 1 includes two peaks at about 18.883° and about 10.889° 2θ. Again, for example, the XRPD spectrum of polymorph A of compound 1 includes two peaks at about 10.889° and about 21.309° 2θ. Again, for example, the XRPD spectrum of polymorph A of compound 1 includes two peaks at about 18.883° and about 21.309° 2θ. In some embodiments, the XRPD spectrum of polymorph A of compound 1 includes all peaks selected from the group consisting of: at about 18.883°, about 10.889°, and about 21.309° 2θ.

[0071] In some embodiments, the XRPD pattern of crystalline form A of Compound 1 comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9) peaks selected from the group consisting of: about 14.974°, about 24.160°, about 18.663°, about 17.109°, about 7.518°, about 20.950°, about 19.602°, about 27.784° and about 28.281° 2θ. For example, the XRPD pattern of crystalline form A of Compound 1 comprises a peak at about 14.974°, about 24.160°, about 18.663°, about 17.109°, about 7.518°, about 20.950°, about 19.602°, about 27.784° or about 28.281° 2θ. In some embodiments, the XRPD pattern of crystalline form A of Compound 1 comprises all of the peaks selected from the group consisting of: about 14.974°, about 24.160°, about 18.663°, about 17.109°, about 7.518°, about 20.950°, about 19.602°, about 27.784° and about 28.281° 2θ.

[0072] In some embodiments, the XRPD pattern of crystalline form A of Compound 1 further comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17) peaks selected from the group consisting of: at about 17.494°, about 14.743°, about 22.386°, about 31.979°, about 21.759°, about 25.017°, about 30.623°, about 10.301°, about 26.760°, about 21.536°, about 19.337°, about 30.103°, about 33.310°, about 31.467°, about 23.309°, about 25.781° and about 27.362° 2θ. For example, the XRPD pattern of crystalline form A of Compound 1 further comprises peaks at about 17.494°, about 14.743°, about 22.386°, about 31.979°, about 21.759°, about 25.017°, about 30.623°, about 10.301°, about 26.760°, about 21.536°, about 19.337°, about 30.103°, about 33.310°, about 31.467°, about 23.309°, about 25.781° or about 27.362° 2θ. Again, for example, the XRPD pattern of crystalline form A of Compound 1 further comprises all peaks selected from the group consisting of: at about 17.494°, about 14.743°, about 22.386°, about 31.979°, about 21.759°, about 25.017°, about 30.623°, about 10.301°, about 26.760°, about 21.536°, about 19.337°, about 30.103°, about 33.310°, about 31.467°, about 23.309°, about 25.781° and about 27.362° 2θ.

[0073] In some embodiments, the XRPD pattern of crystalline form A of Compound 1 comprises all peaks selected from the group consisting of:

[0074] Angle (°2θ) Angle (°2θ) Angle (°2θ) Angle (°2θ) 18.883 14.743 23.309 28.891 10.889 22.386 25.781 34.526 21.309 31.979 27.362 17.884 14.974 21.759 35.751 13.697 24.160 25.017 23.835 32.877 18.663 30.623 23.601 36.134 17.109 10.301 37.946 11.644 7.518 26.760 33.960 37.542 20.950 21.536 39.178 16.364 19.602 19.337 10.569 14.156 27.784 30.103 36.533 35.228 28.281 33.310 26.283 17.494 31.467 29.254

[0075] In some embodiments, the XRPD pattern of crystalline form A of Compound 1 is substantially as Figure 1 shown. In some embodiments, the XRPD pattern of crystalline form A of Compound 1 is as Figure 1 shown.

[0076] In some embodiments, the DSC curve of crystalline form A of Compound 1 comprises an endotherm at about 206.9 °C. In some embodiments, the DSC curve of crystalline form A of Compound 1 is substantially as Figure 3 shown. In some embodiments, the DSC curve of crystalline form A of Compound 1 is as Figure 3 shown.

[0077] III. Polymorph B of Compound 1

[0078] On the other hand, the present disclosure provides polymorph B of Compound 1. In some embodiments, the present disclosure provides substantially pure polymorph B of Compound 1. In some embodiments, Compound 1 is a solvated polymorph, for example, polymorph B of Compound 1 is a methanol solvate.

[0079] In some embodiments, the XRPD pattern of polymorph B of Compound 1 includes one or more (e.g., two, three) peaks selected from the group consisting of: about 6.244°, about 18.679°, and about 12.445° 2θ. For example, the XRPD pattern of polymorph B of Compound 1 includes a peak at about 6.244° 2θ. Again, for example, the XRPD pattern of polymorph B of Compound 1 includes a peak at about 18.679° 2θ. Still again, for example, the XRPD pattern of polymorph B of Compound 1 includes a peak at about 12.445° 2θ. Still again, for example, the XRPD pattern of polymorph B of Compound 1 includes two peaks at about 6.244° and about 18.679° 2θ. Still again, for example, the XRPD pattern of polymorph B of Compound 1 includes two peaks at about 6.244° and about 12.445° 2θ. Still again, for example, the XRPD pattern of polymorph B of Compound 1 includes two peaks at about 18.679° and about 12.445° 2θ. In some embodiments, the XRPD pattern of polymorph B of Compound 1 includes all of the peaks selected from the group consisting of: about 6.244°, about 18.679°, and about 12.445° 2θ.

[0080] In some embodiments, the XRPD pattern of polymorph B of Compound 1 includes one or more (e.g., 2, 3, 4, 5, 6, 7) peaks selected from the group consisting of: about 17.239°, about 12.832°, about 23.559°, about 15.252°, about 22.410°, about 9.344°, and about 25.722° 2θ. For example, the XRPD pattern of polymorph B of Compound 1 includes a peak at about 17.239°, about 12.832°, about 23.559°, about 15.252°, about 22.410°, about 9.344°, or about 25.722° 2θ. In some embodiments, the XRPD pattern of polymorph B of Compound 1 includes all of the peaks selected from the group consisting of: about 17.239°, about 12.832°, about 23.559°, about 15.252°, about 22.410°, about 9.344°, and about 25.722° 2θ.

[0081] In some embodiments, the XRPD pattern of Form B of Compound 1 further comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15) peaks selected from the group consisting of: at about 20.354°, about 16.366°, about 24.731°, about 20.581°, about 28.664°, about 27.044°, about 35.758°, about 17.905°, about 16.087°, about 36.194°, about 34.867°, about 24.984°, about 10.802°, about 29.902° and about 32.584° 2θ. For example, the XRPD pattern of Form B of Compound 1 further comprises peaks at about 20.354°, about 16.366°, about 24.731°, about 20.581°, about 28.664°, about 27.044°, about 35.758°, about 17.905°, about 16.087°, about 36.194°, about 34.867°, about 24.984°, about 10.802°, about 29.902° or about 32.584° 2θ. In some embodiments, the XRPD pattern of Form B of Compound 1 further comprises all of the peaks selected from the group consisting of: at about 20.354°, about 16.366°, about 24.731°, about 20.581°, about 28.664°, about 27.044°, about 35.758°, about 17.905°, about 16.087°, about 36.194°, about 34.867°, about 24.984°, about 10.802°, about 29.902° and about 32.584° 2θ.

[0082] In some embodiments, the XRPD pattern of Form B of Compound 1 comprises all of the peaks selected from the group consisting of:

[0083]

[0084]

[0085] In some embodiments, the XRPD pattern of Form B of Compound 1 is substantially as Figure 5 shown therein. In some embodiments, the XRPD pattern of Form B of Compound 1 is as Figure 5 shown therein.

[0086] In some embodiments, the DSC curve of Form B of Compound 1 comprises endotherms at about 178.3 °C, about 179.7 °C, about 196.6 °C, about 198.7 °C and / or about 203.7 °C. In some embodiments, the DSC curve of Form B of Compound 1 is substantially as Figure 7 shown therein. In some embodiments, the DSC curve of Form B of Compound 1 is as Figure 7as shown in

[0087] IV. Polymorph C of Compound 1

[0088] On the other hand, the present disclosure provides polymorph C of Compound 1. In some embodiments, the present disclosure provides substantially pure polymorph C of Compound 1. In some embodiments, Compound 1 is a solvated polymorph, e.g., polymorph C of Compound 1 is a hydrate.

[0089] In some embodiments, the XRPD pattern of polymorph C of Compound 1 includes one or more (e.g., two, three) peaks selected from the group consisting of: at about 6.656°, about 19.994°, and about 26.732° 2θ. For example, the XRPD pattern of polymorph C of Compound 1 includes a peak at about 6.656° 2θ. Again, for example, the XRPD pattern of polymorph C of Compound 1 includes a peak at about 19.994° 2θ. Still again, for example, the XRPD pattern of polymorph C of Compound 1 includes a peak at about 26.732° 2θ. Still again, for example, the XRPD pattern of polymorph C of Compound 1 includes two peaks at about 6.656° and about 19.994° 2θ. Still again, for example, the XRPD pattern of polymorph C of Compound 1 includes two peaks at about 6.656° and about 26.732° 2θ. Still again, for example, the XRPD pattern of polymorph C of Compound 1 includes two peaks at about 19.994° and about 26.732° 2θ. In some embodiments, the XRPD pattern of polymorph C of Compound 1 includes all of the peaks selected from the group consisting of: at about 6.656°, about 19.994°, and about 26.732° 2θ.

[0090] In some embodiments, the XRPD pattern of polymorph C of Compound 1 includes one or more (e.g., 2, 3, 4, 5, 6, 7) peaks selected from the group consisting of: at about 18.350°, about 20.753°, about 22.176°, about 15.941°, about 20.450°, about 13.320°, and about 17.687° 2θ. For example, the XRPD pattern of polymorph C of Compound 1 includes a peak at about 18.350°, about 20.753°, about 22.176°, about 15.941°, about 20.450°, about 13.320°, or about 17.687° 2θ. In some embodiments, the XRPD pattern of polymorph C of Compound 1 includes all of the peaks selected from the group consisting of: at about 18.350°, about 20.753°, about 22.176°, about 15.941°, about 20.450°, about 13.320°, and about 17.687° 2θ.

[0091] In some embodiments, the XRPD pattern of polymorph C of Compound 1 includes all of the peaks selected from the group consisting of:

[0092] Angle (°2θ) Angle (°2θ) Angle (°2θ) 6.656 20.450 12.470 19.994 13.320 3.724 26.732 17.687 25.036 18.350 19.205 30.741 20.753 23.302 4.454 22.176 9.183 15.941 8.700

[0093] In some embodiments, the XRPD pattern of crystalline form C of Compound 1 is substantially as shown in Figure 9 as shown therein. In some embodiments, the XRPD pattern of crystalline form C of Compound 1 is as shown in Figure 9 as shown therein.

[0094] In some embodiments, the DSC curve of crystalline form C of Compound 1 is substantially as shown in Figure 11 as shown therein. In some embodiments, the DSC curve of crystalline form C of Compound 1 is as shown in Figure 11 as shown therein.

[0095] V. Polymorph D of Compound 1

[0096] On the other hand, the present disclosure provides crystalline form D of Compound 1. In some embodiments, the present disclosure provides substantially pure crystalline form D of Compound 1. In some embodiments, Compound 1 is an unsolvated crystalline form, for example, crystalline form D of Compound 1 is an anhydrate.

[0097] In some embodiments, the XRPD pattern of crystalline form D of Compound 1 includes one or more (e.g., two, three) peaks selected from the group consisting of: at about 16.956°, about 5.883°, and about 22.181° 2θ. For example, the XRPD pattern of crystalline form D of Compound 1 includes a peak at about 16.956° 2θ. Again, for example, the XRPD pattern of crystalline form D of Compound 1 includes a peak at about 5.883° 2θ. Still again, for example, the XRPD pattern of crystalline form D of Compound 1 includes a peak at about 22.181° 2θ. Still again, for example, the XRPD pattern of crystalline form D of Compound 1 includes two peaks at about 16.956° and about 5.883° 2θ. Still again, for example, the XRPD pattern of crystalline form D of Compound 1 includes two peaks at about 16.956° and about 22.181° 2θ. Still again, for example, the XRPD pattern of crystalline form D of Compound 1 includes two peaks at about 5.883° and about 22.181° 2θ. In some embodiments, the XRPD pattern of crystalline form D of Compound 1 includes all of the peaks selected from the group consisting of: at about 16.956°, about 5.883°, and about 22.181° 2θ.

[0098] In some embodiments, the XRPD pattern of crystalline form D of Compound 1 includes one or more (e.g., 2, 3, 4, 5, 6) peaks selected from the group consisting of: at about 17.663°, about 13.291°, about 22.909°, about 20.718°, about 26.900°, and about 18.113° 2θ. For example, the XRPD pattern of crystalline form D of Compound 1 includes peaks at about 17.663°, about 13.291°, about 22.909°, about 20.718°, about 26.900°, or about 18.113° 2θ. In some embodiments, the XRPD pattern of crystalline form D of Compound 1 includes all of the peaks selected from the group consisting of: at about 17.663°, about 13.291°, about 22.909°, about 20.718°, about 26.900°, or about 18.113° 2θ.

[0099] In some embodiments, the XRPD pattern of crystalline form D of Compound 1 further includes one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15) peaks selected from the group consisting of: at about 20.205°, about 11.727°, about 21.756°, about 22.617°, about 20.956°, about 29.635°, about 24.990°, about 12.566°, about 10.080°, about 24.078°, about 9.761°, about 19.581°, about 27.280°, about 25.637°, and about 12.754° 2θ. For example, the XRPD pattern of crystalline form D of Compound 1 further includes peaks at about 20.205°, about 11.727°, about 21.756°, about 22.617°, about 20.956°, about 29.635°, about 24.990°, about 12.566°, about 10.080°, about 24.078°, about 9.761°, about 19.581°, about 27.280°, about 25.637°, or about 12.754° 2θ. In some embodiments, the XRPD pattern of crystalline form D of Compound 1 further includes all of the peaks selected from the group consisting of: at about 20.205°, about 11.727°, about 21.756°, about 22.617°, about 20.956°, about 29.635°, about 24.990°, about 12.566°, about 10.080°, about 24.078°, about 9.761°, about 19.581°, about 27.280°, about 25.637°, and about 12.754° 2θ.

[0100] In some embodiments, the XRPD pattern of crystalline form D of Compound 1 includes all of the peaks selected from the group consisting of:

[0101] Angle (°2θ) Angle (°2θ) Angle (°2θ) Angle (°2θ) 16.956 21.756 25.637 39.200 5.883 22.617 12.754 33.344 22.181 20.956 14.335 18.891 17.663 29.635 35.599 34.841 13.291 24.990 10.411 10.830 22.909 12.566 30.457 28.859 20.718 10.080 11.067 14.954 26.900 24.078 19.175 28.254 18.113 9.761 34.373 31.083 20.205 19.581 36.486 7.456 11.727 27.280 37.185

[0102] In some embodiments, the XRPD pattern of crystalline form D of Compound 1 is substantially as Figure 13 shown in. In some embodiments, the XRPD pattern of crystalline form D of Compound 1 is as Figure 13 shown in.

[0103] In some embodiments, the DSC curve of crystalline form D of Compound 1 includes an endotherm at about 207.4 °C. In some embodiments, the DSC curve of crystalline form D of Compound 1 is substantially as Figure 15 shown in. In some embodiments, the DSC curve of crystalline form D of Compound 1 is as Figure 15 shown in.

[0104] VI. Polymorph E of Compound 1

[0105] On the other hand, the present disclosure provides crystalline form E of Compound 1. In some embodiments, the present disclosure provides substantially pure crystalline form E of Compound 1. In some embodiments, Compound 1 is an unsolvated crystalline form, for example, crystalline form E of Compound 1 is an anhydrate.

[0106] In some embodiments, the XRPD pattern of crystalline form E of Compound 1 includes one or more (e.g., two, three) peaks selected from the group consisting of: at about 18.089°, about 26.315°, and about 19.775° 2θ. For example, the XRPD pattern of crystalline form E of Compound 1 includes a peak at about 18.089° 2θ. Again, for example, the XRPD pattern of crystalline form E of Compound 1 includes a peak at about 26.315° 2θ. Still again, for example, the XRPD pattern of crystalline form E of Compound 1 includes a peak at about 19.775° 2θ. Still again, for example, the XRPD pattern of crystalline form E of Compound 1 includes two peaks at about 18.089° and about 26.315° 2θ. Still again, for example, the XRPD pattern of crystalline form E of Compound 1 includes two peaks at about 18.089° and about 19.775° 2θ. Still again, for example, the XRPD pattern of crystalline form E of Compound 1 includes two peaks at about 26.315° and about 19.775° 2θ. In some embodiments, the XRPD pattern of crystalline form E of Compound 1 includes all of the peaks selected from the group consisting of: at about 18.089°, about 26.315°, and about 19.775° 2θ.

[0107] In some embodiments, the XRPD pattern of crystalline form E of Compound 1 includes one or more (e.g., 2, 3, 4, 5) peaks selected from the group consisting of: at about 24.177°, about 20.803°, about 16.018°, about 21.429°, and about 14.608° 2θ. The XRPD pattern of crystalline form E of Compound 1 includes a peak at about 24.177°, about 20.803°, about 16.018°, about 21.429°, or about 14.608° 2θ. In some embodiments, the XRPD pattern of crystalline form E of Compound 1 includes all of the peaks selected from the group consisting of: at about 24.177°, about 20.803°, about 16.018°, about 21.429°, and about 14.608° 2θ.

[0108] In some embodiments, the XRPD pattern of crystalline form E of Compound 1 includes one or more (e.g., 2, 3, 4, 5) peaks selected from the group consisting of: at about 21.314°, about 7.794°, about 20.007°, about 8.744°, and about 15.573° 2θ. For example, the XRPD pattern of crystalline form E of Compound 1 includes a peak at about 21.314°, about 7.794°, about 20.007°, about 8.744°, or about 15.573° 2θ. In some embodiments, the XRPD pattern of crystalline form E of Compound 1 includes all of the peaks selected from the group consisting of: at about 21.314°, about 7.794°, about 20.007°, about 8.744°, and about 15.573° 2θ.

[0109] In some embodiments, the XRPD pattern of crystalline form E of Compound 1 includes all of the peaks selected from the group consisting of:

[0110] Angle (°2θ) Angle (°2θ) Angle (°2θ) Angle (°2θ) 18.089 18.636 27.764 29.879 26.315 28.686 17.267 34.904 19.775 16.43 14.962 32.851 24.177 25.182 18.864 35.467 20.803 20.584 12.049 31.997 16.018 10.856 17.085 12.878 21.429 22.515 7.486 11.239 14.608 19.611 17.477 30.606 21.314 14.233 13.613 3.691 7.794 22.83 36.59 34.151 20.007 23.557 28.293 33.397 8.744 27.166 21.743 15.573 24.647 31.524

[0111] In some embodiments, the XRPD pattern of crystalline form E of Compound 1 is substantially as Figure 17 shown. In some embodiments, the XRPD pattern of crystalline form E of Compound 1 is as Figure 17 shown.

[0112] VII. Polymorph A of L - malate of Compound 1

[0113] On the other hand, the present disclosure provides crystalline form A of the malate salt of Compound 1. In some embodiments, the present disclosure provides crystalline form A of the L-malate salt of Compound 1.

[0114] In some embodiments, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes one or more (e.g., two, three) peaks selected from the group consisting of: at about 19.349°, about 22.304°, and about 20.078° 2θ. For example, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes a peak at about 19.349° 2θ. Again, for example, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes a peak at about 22.304° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes a peak at about 20.078° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes two peaks at about 19.349° and about 22.304° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes two peaks at about 19.349° and about 20.078° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes two peaks at about 22.304° and about 20.078° 2θ. In some embodiments, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes all of the peaks selected from the group consisting of: at about 19.349°, about 22.304°, and about 20.078° 2θ.

[0115] In some embodiments, the XRPD pattern of crystalline form A of the L-malate of Compound 1 further includes one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) peaks selected from the group consisting of: at about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597°, and about 17.581° 2θ. For example, the XRPD pattern of crystalline form A of the L-malate of Compound 1 includes a peak at about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597°, or about 17.581° 2θ. In some embodiments, the XRPD pattern of crystalline form A of the L-malate of Compound 1 further includes all of the peaks selected from the group consisting of: at about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597°, and about 17.581° 2θ.

[0116] In some embodiments, the XRPD pattern of crystalline form A of the L-malate salt of Compound 1 further comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17) peaks selected from the group consisting of: about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704°, and about 21.715° 2θ. For example, the XRPD pattern of crystalline form A of the L-malate salt of Compound 1 further comprises a peak at about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704°, or about 21.715° 2θ. As another example, the XRPD pattern of crystalline form A of the L-malate salt of Compound 1 further comprises all of the peaks selected from the group consisting of: about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704°, and about 21.715° 2θ.

[0117] In some embodiments, the XRPD pattern of Form A of the L-malate salt of Compound 1 comprises all peaks selected from the group consisting of: at about 19.349°, about 22.304°, about 20.078°, about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597°, about 17.581°, about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704°, about 21.715°, about 36.432°, about 31.492°, about 27.976°, about 37.328°, about 17.975°, about 38.681°, about 15.886°, about 13.678°, about 32.223°, about 26.405°, about 27.514°, about 31.082°, about 14.561°, about 39.38°, about 35.442°, about 26.035°, about 26.739°, about 35.592°, about 11.409°, about 10.2°, about 8.962°, about 34.334°, about 28.307°, about 35.984°, about 29.228°, about 39.705°, about 33.715° and about 30.135° 2θ.

[0118] In some embodiments, the XRPD pattern of Form A of the L-malate salt of Compound 1 is substantially as Figure 18 shown therein. In some embodiments, the XRPD pattern of Form A of the L-malate salt of Compound 1 is as Figure 18 shown therein.

[0119] In some embodiments, the DSC curve of Form A of the L-malate salt of Compound 1 comprises an endotherm at about 160.3 °C. In some embodiments, the DSC curve of Form A of the L-malate salt of Compound 1 is substantially as Figure 20 shown therein. In some embodiments, the DSC curve of Form A of the L-malate salt of Compound 1 is as Figure 20 shown therein.

[0120] VIII. Polymorph A of L - tartrate of Compound 1

[0121] On the other hand, the present disclosure provides the tartrate salt of Form A of Compound 1. In some embodiments, the present disclosure provides Form A of the L-tartrate salt of Compound 1.

[0122] In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes one or more (e.g., two, three) peaks selected from the group consisting of: at about 18.564°, about 21.685°, and about 18.774° 2θ. For example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes a peak at about 18.564° 2θ. Again, for example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes a peak at about 21.685° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes a peak at about 18.774° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes two peaks at about 18.564° and about 21.685° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes two peaks at about 21.685° and about 18.774° 2θ. Still again, for example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes two peaks at about 18.564° and about 18.774° 2θ. In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes all of the peaks selected from the group consisting of: at about 18.564°, about 21.685°, and about 18.774° 2θ.

[0123] In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes one or more (e.g., 2, 3, 4, 5, 6) peaks selected from the group consisting of: at about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965°, and about 24.193° 2θ. For example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes a peak at about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965°, or about 24.193° 2θ. In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 includes all of the peaks selected from the group consisting of: at about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965°, and about 24.193° 2θ.

[0124] In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 further comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18) peaks selected from the group consisting of: about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641°, and about 19.08° 2θ. For example, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 further comprises a peak at about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641°, or about 19.08° 2θ. In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 further comprises all of the peaks selected from the group consisting of: about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641°, and about 19.08° 2θ.

[0125] In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 further comprises all peaks selected from the group consisting of: at about 18.564°, about 21.685°, about 18.774°, about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965°, about 24.193°, about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641°, about 19.08°, about 10.165°, about 33.761°, about 29.071°, about 25.916°, about 16.943°, about 32.967°, about 23.691°, about 22.788°, about 34.232°, about 35.17°, about 34.812°, about 27.271°, about 32.24°, about 39.082°, about 30.774°, about 15.453°, about 30.534°, about 31.183°, about 37.181°, about 35.805°, about 28.554°, about 29.645°, about 39.789°, about 38.258°, about 10.432°, about 13.968° and about 36.619° 2θ.

[0126] In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 is substantially as Figure 22 shown therein. In some embodiments, the XRPD pattern of crystalline form A of the L-tartrate salt of Compound 1 is as Figure 22 shown therein.

[0127] In some embodiments, the DSC curve of crystalline form A of the L-tartrate salt of Compound 1 comprises an endotherm at about 189.9 °C. In some embodiments, the DSC curve of crystalline form A of the L-tartrate salt of Compound 1 is substantially as Figure 24 shown therein. In some embodiments, the DSC curve of crystalline form A of the L-tartrate salt of Compound 1 is as Figure 24 shown therein.

[0128] IX. Pharmaceutical Composition

[0129] On the other hand, the present disclosure provides a pharmaceutical composition comprising Compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, wherein Compound 1 includes crystal forms selected from the group consisting of: Crystal Form A, Crystal Form B, Crystal Form C, Crystal Form D, Crystal Form E of Compound 1 described herein, Crystal Form A of L-malate of Compound 1, and Crystal Form A of L-tartrate of Compound 1.

[0130] As used herein, a pharmaceutical composition refers to a mixture of one or more crystal forms of Compound 1 or a pharmaceutically acceptable salt thereof described herein with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. In some embodiments, the pharmaceutical composition facilitates the administration of one or more crystal forms of Compound 1 or a pharmaceutically acceptable salt thereof described herein to an organism. In some embodiments, practicing the therapeutic methods or uses provided herein includes administering or using a pharmaceutical composition comprising a therapeutically effective amount of a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof described herein. In a specific embodiment, the therapeutic methods provided herein include administering such a pharmaceutical composition to a mammal suffering from a disease or disorder to be treated. In one embodiment, the mammal is a human. In some embodiments, the therapeutically effective amount varies widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. In various embodiments, the crystal forms of Compound 1 or a pharmaceutically acceptable salt thereof described herein are used alone or in combination with one or more additional therapeutic agents as components of a mixture.

[0131] In some embodiments, the pharmaceutical composition is formulated in any manner, including using one or more physiologically acceptable carriers, which carriers comprise excipients and / or adjuvants that facilitate the processing of one or more crystal forms of Compound 1 or a pharmaceutically acceptable salt thereof described herein into a pharmaceutical formulation. In some embodiments, the suitable formulation depends on the selected route of administration. In various embodiments, any technique, carrier, and excipient are used appropriately.

[0132] In some embodiments, the composition is formulated in a formulation for topical or systemic delivery. Examples of such formulations are formulations for oral administration, injection, topical application, pulmonary administration, or implantation.

[0133] In some embodiments, the pharmaceutical composition provided herein is formulated for intravenous injection. In certain aspects, the intravenous injection formulation provided herein is formulated as an aqueous solution and, in some embodiments, is formulated in a physiologically compatible buffer such as Hank's solution, Ringer's solution, or physiological saline buffer.

[0134] In certain embodiments, the pharmaceutical composition provided herein is formulated for transmucosal administration. In some embodiments, the transmucosal formulation includes a penetrant suitable for the barrier to be penetrated.

[0135] In certain embodiments, the pharmaceutical compositions provided herein are formulated for other parenteral injections. Suitable formulations include aqueous or non-aqueous solutions and, in one embodiment, have a physiologically compatible buffer or excipient.

[0136] In some embodiments, parenteral injections include bolus or infusion. In some embodiments, the formulations for injection are presented in unit dosage forms, such as in ampoules or in multi-dose containers, and contain a preservative. In some embodiments, the pharmaceutical compositions described herein are in a form suitable for parenteral injection, which is a sterile suspension, solution or emulsion in an oily or aqueous vehicle and optionally contains formulating agents such as suspending agents, stabilizers and / or dispersing agents. Pharmaceutical preparations for parenteral administration contain an aqueous solution of the active ingredient in a water-soluble form. In some embodiments, a suspension of the active ingredient is prepared as a suitable oily injection suspension. Suitable lipophilic solvents or vehicles include fatty oils (such as sesame oil) or synthetic fatty acid esters (such as ethyl oleate or triglycerides) or liposomes. In some embodiments, the aqueous injection suspension contains substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension also contains suitable stabilizers or formulating agents that increase the solubility of the compound to allow the preparation of a highly concentrated solution. In alternative embodiments, the active ingredient is in powder form for formulation with a suitable vehicle such as sterile pyrogen-free water before use.

[0137] In some embodiments, the pharmaceutical compositions provided herein are formulated for oral administration. In certain aspects, the oral formulations provided herein contain one or more crystal forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof, which are formulated together with a pharmaceutically acceptable carrier or excipient. Such carriers enable the crystal forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof to be formulated into tablets, powders, pills, dragees, capsules, oral solutions, gels, syrups, elixirs, suspensions, etc. for oral ingestion by the patient to be treated.

[0138] In some embodiments, a pharmaceutical composition for oral use is obtained by mixing one or more solid excipients with one or more crystalline forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof, optionally grinding the resulting mixture, and, if desired, processing the mixture of granules after addition of suitable auxiliaries to obtain tablets or dragee cores. Suitable excipients specifically include fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. If desired, disintegrants are optionally added, such as cross-linked sodium carboxymethylcellulose, polyvinylpyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate.

[0139] In some embodiments, provided herein is a pharmaceutical composition formulated as a dragee core having a suitable coating. In some embodiments, a concentrated sugar solution is used to form the suitable coating and optionally contains gum arabic, talc, polyvinylpyrrolidone, carbomer gel, polyethylene glycol and / or titanium dioxide, lacquer solution and a suitable organic solvent or solvent mixture. In some embodiments, dyes and / or pigments are added to the tablets, dragees and / or their coatings for, for example, identifying or characterizing different combinations of active compound dosages.

[0140] In some embodiments, the oral pharmaceutical compositions used include push-fit capsules made of gelatin and soft-sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. In some embodiments, the push-fit capsules may contain active ingredients, which are mixed with fillers such as lactose, binders such as starch and / or lubricants such as talc or magnesium stearate and optionally stabilizers. In some embodiments, in the soft capsules, the active ingredients are dissolved or suspended in a suitable liquid such as fatty oil, liquid paraffin or liquid polyethylene glycol. Additionally, stabilizers are optionally added. In some or any embodiments, the preparations for oral administration are in a dosage suitable for such administration.

[0141] In some embodiments, the pharmaceutical compositions provided herein are formulated for buccal or sublingual administration. In some embodiments, the buccal or sublingual compositions are in the form of tablets, lozenges or gels formulated in a conventional manner.

[0142] In some embodiments, the pharmaceutical compositions provided herein are formulated for transdermal administration. In some embodiments, administration of such compositions is by means of a transdermal delivery device and a transdermal delivery patch. In some or any embodiments, the composition is a lipophilic emulsion or buffered aqueous solution dissolved and / or dispersed in a polymer or adhesive. Such patches can include patches configured for continuous, pulsed, or on-demand delivery of the medicament. In some embodiments, transdermal delivery of one or more crystalline forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof is accomplished by using an iontophoretic patch or the like. In some or any embodiments, the transdermal patch provides controlled delivery of one or more crystalline forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof. In some embodiments, the rate of absorption is reduced by using a rate-controlling membrane or by trapping the compound in a polymer matrix or gel. Conversely, absorption enhancers are optionally used to increase absorption. Absorption enhancers and carriers include pharmaceutically acceptable solvents that are absorbable and assist the compound in passing through the skin. For example, the transdermal device is in the form of a bandage that includes a backing member, a reservoir containing one or more crystalline forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof and optionally a carrier, an optional rate-controlling barrier for delivering the compound to the host skin at a controlled and predetermined rate over a long period of time, and means for securing the device to the skin.

[0143] In some or any embodiments, the pharmaceutical compositions provided herein are formulated for administration by inhalation. In some embodiments, in such pharmaceutical compositions formulated for inhalation, one or more crystalline forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof are in the form of an aerosol, a mist, or a powder. In some embodiments, a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas) is used to conveniently deliver the pharmaceutical composition described herein in the form of an aerosol spray from a pressurized pack or nebulizer. In certain aspects of pressurized aerosols, the dosage unit is determined by providing a valve for delivering a metered amount. In some or any embodiments, capsules and cartridges, such as (by way of example only) gelatin, formulated for an inhaler or insufflator contain a powder mixture of one or more crystalline forms of Compound 1 described herein or a pharmaceutically acceptable salt thereof and a suitable powder matrix (such as lactose or starch).

[0144] In some embodiments, the compounds described herein are formulated in rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, gelatin suppositories, or retention enemas. In some embodiments, the rectal compositions optionally contain conventional suppository bases such as cocoa butter or other glycerides and synthetic polymers such as polyvinylpyrrolidone, PEG, etc. In the suppository form of some or any compositions, a low melting point wax such as, but not limited to, a mixture of fatty acid glycerides is first melted, optionally in combination with cocoa butter.

[0145] Methods for preparing a composition containing a compound described herein include formulating the compound with one or more inert pharmaceutically acceptable excipients or carriers to form a solid, semi-solid or liquid. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which the compound is dissolved, emulsions containing the compound, or solutions containing liposomes, micelles or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. In various embodiments, the composition is in the form of a liquid solution or suspension, a solid form adapted to be dissolved or suspended in a liquid prior to use, or as an emulsion. These compositions optionally contain small amounts of non-toxic auxiliary substances such as wetting agents or emulsifying agents, pH buffering agents, and the like.

[0146] In some embodiments, a composition containing a compound described herein takes the form of a liquid, wherein the agent is present in a solution, a suspension, or both. In some embodiments, when the composition is administered as a solution or suspension, a first portion of the agent is present in the solution and a second portion of the agent is present in suspension in particulate form in the liquid matrix. In some embodiments, the liquid composition includes a gel formulation. In other embodiments, the liquid composition is aqueous.

[0147] Useful aqueous suspensions optionally contain one or more polymers as suspending agents. Useful polymers include water-soluble polymers such as cellulose polymers, e.g., hydroxypropylmethylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Useful compositions optionally contain a mucoadhesive polymer selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0148] Useful compositions optionally include solubilizing agents to aid in the solubility of the compounds described herein. The term "solubilizing agent" generally includes agents that cause the formation of a micellar solution or true solution of the agent. Solubilizing agents include certain acceptable nonionic surfactants, such as polysorbate 80, and ophthalmically acceptable diols, polyethylene glycols, e.g., polyethylene glycol 400 and ethylene glycol ethers.

[0149] Useful compositions optionally include one or more pH regulators or buffering agents, including acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and tromethamine; and buffers such as citrate / glucose, sodium bicarbonate, and ammonium chloride. The amounts of such acids, bases, and buffers are those required to maintain the pH of the composition within an acceptable range.

[0150] The useful composition optionally includes an amount of one or more salts sufficient to bring the osmotic pressure of the composition into an acceptable range. Such salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate.

[0151] Certain useful pharmaceutical compositions optionally include one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as phenylmercuric borate and thimerosal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide and cetylpyridinium chloride.

[0152] Some useful compositions optionally include one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils such as polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkyl ethers and alkyl phenyl ethers such as octylphenol polyether-10 and octylphenol polyether-40.

[0153] Certain useful compositions optionally include one or more antioxidants to enhance chemical stability when needed. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0154] In some embodiments, the aqueous suspension composition is packaged in a single-dose non-resealable container. In alternative embodiments, a multi-dose resealable container is used, in which case a preservative is typically included in the composition.

[0155] In various embodiments, any delivery system for hydrophobic pharmaceutical compounds is used. Liposomes and emulsions are examples of hydrophobic drug delivery vehicles or carriers. In some or any embodiments, certain organic solvents such as N-methylpyrrolidone are used. In some embodiments, a sustained-release system such as a semipermeable matrix of a solid hydrophobic polymer containing a therapeutic agent is used to deliver the compound. A variety of sustained-release materials are used in the embodiments herein. In some or any embodiments, a sustained-release capsule releases the compound for periods ranging from several weeks to over 100 days. In some embodiments, additional protein stabilization strategies are used depending on the chemical nature and biological stability of the therapeutic agent.

[0156] In some or any embodiments, the formulations or compositions described herein benefit from and / or optionally contain metal chelators, thiol-containing compounds, and other common stabilizers. Examples of such stabilizers include, but are not limited to: (a) about 0.5% to about 2% w / v glycerol, (b) about 0.1% to about 1% w / v methionine, (c) about 0.1% to about 2% monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w / w ascorbic acid, (f) 0.003% to about 0.02% w / v polysorbate 80, (g) 0.001% to about 0.05% w / v polysorbate 20, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentosan polysulfate and other heparinoids, (m) divalent cations such as magnesium and zinc; or (n) combinations thereof.

[0157] In some or any embodiments, the pharmaceutical compositions described herein are unit dosage forms suitable for single administration of an exact dose. In some cases, within the unit dosage form, the formulation is divided into unit doses comprising an appropriate amount of one or more compounds. In some or any embodiments, a unit dose is a packaged form containing a discrete amount of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. In some embodiments, an aqueous suspension composition is packaged in a single-dose non-reclosable container. In alternative embodiments, a multi-dose reclosable container is used, in which case a preservative is typically included in the composition. By way of example only, in some embodiments, a formulation for parenteral injection is in unit dosage form, which includes, but is not limited to, an ampoule, or in a multi-dose container with a preservative added.

[0158] X. Use

[0159] On the other hand, the present disclosure also provides the use of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or pharmaceutical compositions in the preparation of a medicament for:

[0160] (a) preventing or treating a disease, disorder or condition associated with abnormal activity / level of the enzyme GCS;

[0161] (b) preventing or treating a GCS-mediated disease, disorder or condition;

[0162] (c) inhibiting the activity / level of the enzyme GCS; or

[0163] (d) non-therapeutically inhibiting the activity / level of the GCS enzyme in vitro.

[0164] The phrase "GCS-mediated disease, disorder or condition" refers to a disease, disorder or condition associated with an abnormal increase or decrease in the activity / level of a GCS enzyme that may be caused by abnormal activation or inactivation of the GCS enzyme in a subject. In some embodiments, the GCS-mediated disease, disorder or condition is a disease, disorder or condition associated with an abnormal increase in the activity / level of the GCS enzyme.

[0165] In some embodiments, the disease, disorder or condition associated with the abnormal activity / level of the enzyme GCS, or the GCS-mediated disease, disorder or condition, is selected from the group consisting of: glycolipid storage diseases (e.g., Tay-Sachs disease, Sandhoff disease, GM1 gangliosidosis and Fabry disease); diseases associated with glycolipid accumulation (e.g., Gaucher disease); diseases causing renal hypertrophy or hyperplasia, such as diabetic nephropathy; diseases causing hyperglycemia or hyperinsulinemia; cancers with abnormal glycolipid synthesis; infectious diseases caused by organisms that use cell surface glycolipids as receptors or for which the synthesis of glucosylceramide is essential or important; metabolic disorders, such as atherosclerosis, polycystic kidney disease, renal hypertrophy and diabetes; obesity; cancers, such as breast cancer, renal adenocarcinoma, brain cancer, neuroblastoma, lung cancer, intestinal cancer, pancreatic cancer and prostate cancer; neuronal disorders; neuronal injury; inflammatory diseases or disorders (e.g., rheumatoid arthritis, Crohn's disease, asthma and sepsis); pain (e.g., neuropathic pain, inflammatory pain, headache, somatic pain, visceral pain, referred pain); cognitive disorders (e.g., agnosia; amnesia; aphasia; apraxia; delirium; dementia, including AIDS dementia complex, Binswanger's disease, Lewy body dementia, frontotemporal dementia, mild cognitive impairment, multi-infarct dementia, Pick's disease, semantic dementia, Alzheimer's disease and vascular dementia; and learning disorders, including Asperger syndrome, attention deficit disorder, attention deficit hyperactivity disorder, autism, childhood disintegrative disorder and Rett syndrome); neurodegenerative disorders (such as Alzheimer's disease, corticobasal degeneration, Creutzfeldt-Jakob disease, frontotemporal lobar degeneration, Huntington's disease, multiple sclerosis, normal pressure hydrocephalus, organic chronic brain syndrome, Parkinson's disease, Pick's disease, progressive supranuclear palsy and Alzheimer's (Alzheimer type), glomerular diseases) and non-alcoholic fatty liver disease (NALD). In some embodiments, the disease, disorder or condition is Tay-Sachs disease, Sandhoff disease, GM1 gangliosidosis, Fabry disease, Gaucher disease or polycystic kidney disease.

[0166] On the other hand, the present disclosure also provides a method for preventing or treating a GCS-mediated disease, disorder, or condition in a subject, the method comprising administering to the subject polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., polymorph A of the L-malate salt of compound 1, polymorph A of the L-tartrate salt of compound 1), or a pharmaceutical composition.

[0167] On the other hand, the present disclosure also provides polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., polymorph A of the L-malate salt of compound 1, polymorph A of the L-tartrate salt of compound 1), or a pharmaceutical composition for preventing or treating a GCS-mediated disease, disorder, or condition.

[0168] On the other hand, the present disclosure also provides a method for inhibiting GCS activity, the method comprising administering to a subject in need an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., polymorph A of the L-malate salt of compound 1, polymorph A of the L-tartrate salt of compound 1), or a pharmaceutical composition.

[0169] XI. Administration Method and Treatment Regimen

[0170] In any of the foregoing embodiments, additional embodiments include administering a single effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., polymorph A of the L-malate salt of compound 1, polymorph A of the L-tartrate salt of compound 1), or a pharmaceutical composition, including additional embodiments wherein polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., polymorph A of the L-malate salt of compound 1, polymorph A of the L-tartrate salt of compound 1), or a pharmaceutical composition is administered to the subject (i) once; (ii) multiple times within a day; (iii) continuously; or (iv) sequentially.

[0171] In any of the foregoing embodiments, additional embodiments are enteral administration, parenteral administration, or both, and wherein:

[0172] (a) An effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., polymorph A of the L-malate salt of compound 1, polymorph A of the L-tartrate salt of compound 1), or a pharmaceutical composition is administered systemically to the subject;

[0173] (b) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or a pharmaceutical composition orally to a subject;

[0174] (c) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or a pharmaceutical composition intravenously to a subject;

[0175] (d) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or a pharmaceutical composition by inhalation to a subject;

[0176] (e) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or a pharmaceutical composition by intranasal administration to a subject;

[0177] (f) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or a pharmaceutical composition by injection to a subject;

[0178] (g) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts or a pharmaceutical composition topically (to the skin) to a subject;

[0179] (h) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or a pharmaceutical composition by ocular administration to a subject; and / or

[0180] (i) Administering an effective amount of polymorph A, polymorph B, polymorph C, polymorph D, polymorph E of compound 1 described herein, its pharmaceutically acceptable salts (e.g., polymorph A of L-malate of compound 1, polymorph A of L-tartrate of compound 1) or a pharmaceutical composition rectally to a subject.

[0181] In some or any embodiments, a crystalline form of Compound 1 described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition is administered for prophylactic and / or therapeutic treatment. In some or any therapeutic applications, a crystalline form of Compound 1 described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition is administered to a patient already suffering from a disease or condition in an amount sufficient to cure or at least partially inhibit the symptoms of the disease or condition. In some embodiments, the effective amount for this use will depend on the severity and course of the disease or condition, previous therapy, the health status, weight, and response to the drug of the patient, as well as the judgment of the attending physician. In certain cases, it is considered appropriate for the caregiver to determine such therapeutically effective amounts through routine experimentation, including but not limited to dose escalation clinical trials.

[0182] In some or any prophylactic applications, a crystalline form of Compound 1 described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition is administered to a patient who is susceptible to a particular disease, disorder, or condition or who is otherwise at risk of a particular disease, disorder, or condition. In some embodiments, the amount administered is defined as a "prophylactically effective amount or dose". In some or any embodiments of this use, the exact amount of the crystalline form of Compound 1 described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition to be administered depends on the health status, weight, etc. of the patient. In some embodiments, it is considered appropriate for the caregiver to determine such prophylactically effective amounts through routine experimentation (e.g., dose escalation clinical trials). In some or any embodiments, when used in a patient, the effective amount for this use will depend on the severity and course of the disease, disorder, or condition, previous therapy, the health status of the patient, and response to the drug, as well as the judgment of the attending physician.

[0183] In some or any embodiments, additional embodiments include multiple administrations of an effective amount of the compound, including additional embodiments wherein:

[0184] (a) A crystalline form A, B, C, D, or E of Compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., crystalline form A of the L-malate salt of Compound 1, crystalline form A of the L-tartrate salt of Compound 1), or a pharmaceutical composition is administered as a single dose;

[0185] (b) Multiple administrations are spaced 6 hours apart; or

[0186] (c) A crystalline form A, B, C, D, or E of Compound 1 described herein, a pharmaceutically acceptable salt thereof (e.g., crystalline form A of the L-malate salt of Compound 1, crystalline form A of the L-tartrate salt of Compound 1), or a pharmaceutical composition is administered to the subject every 8 hours.

[0187] In certain cases, after administration of the crystalline forms of Compound 1 described herein, their pharmaceutically acceptable salts, or pharmaceutical compositions, the condition of the patient does not improve or does not improve significantly, and, at the discretion of the physician, the crystalline forms of Compound 1 described herein, their pharmaceutically acceptable salts, or pharmaceutical compositions are optionally administered chronically, i.e., over an extended period of time, including throughout the patient's life cycle, to improve or otherwise control or limit the symptoms of the patient's disease or condition.

[0188] In certain cases where the patient's condition does improve or does not improve significantly, at the discretion of the physician, administration of the crystalline forms of Compound 1 described herein, their pharmaceutically acceptable salts, or pharmaceutical compositions is optionally continued; alternatively, the dose of the administered drug is optionally temporarily reduced or temporarily discontinued for a specific length of time (i.e., a "drug holiday"). In some or any embodiments, the length of the drug holiday varies between 2 days and 1 year, by way of example only, including 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. Dose reductions during the drug holiday include reductions from about 10% to about 100%, by way of example only, including about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0189] In some or any embodiments, once the patient's condition has improved, a maintenance dose is administered as necessary. In some embodiments, the administration of, for example, the maintenance dose, or the frequency of administration, or both, is reduced, according to the symptoms, to a level that maintains the improved disease, disorder, or condition. However, in some or any embodiments, upon any recurrence of symptoms, the patient is optionally given long-term intermittent treatment.

[0190] In some or any embodiments, the amount corresponding to an effective amount of a given agent varies depending on factors such as the specific compound, the disease or condition and its severity, and the identity of the subject or host to be treated (e.g., body weight). However, in some embodiments, the effective amount is determined based on the specific circumstances surrounding the case, including, for example, the specific agent being administered, the route of administration, the condition being treated, and the subject or host being treated. However, in some or any embodiments, the dosage for adult treatment ranges from about 0.02 mg / day to about 5000 mg / day, and in specific embodiments is from about 1 mg / day to about 1500 mg / day. In various embodiments, the desired dosage is conveniently provided in a single-dose form or in divided-dose forms administered simultaneously (or within a short time period) or at appropriate intervals, such as two, three, four or more divided doses per day.

[0191] In some or any embodiments, the daily dosage applicable to the crystalline form of Compound 1 described herein, its pharmaceutically acceptable salts or pharmaceutical compositions is from about 0.01 mg / kg body weight to about 2.5 mg / kg body weight. In some embodiments, the indicated daily dosage for larger subjects (including but not limited to humans) ranges from about 0.5 mg to about 100 mg, and is conveniently administered in divided doses, including but not limited to up to four times per day or in a sustained-release form. In some or any embodiments, suitable unit dosage forms for oral administration contain from about 1 mg to about 50 mg of the active ingredient. The foregoing ranges are merely suggestive, since the number of variables regarding an individual treatment regimen is large and fairly large deviations from these recommended values are not uncommon. In some or any embodiments, the dosage varies according to many variables, which are not limited to the activity of the compound used, the disease or condition to be treated, the mode of administration, the needs of the individual subject, the severity of the disease or condition being treated, and the judgment of the practitioner.

[0192] In some or any embodiments, the toxicity and therapeutic efficacy of such treatment regimens are determined by standard pharmaceutical procedures in cell cultures or experimental animals, including but not limited to LD 50 (the lethal dose for 50% of the population) and ED 50 (the dose effective in treating 50% of the population) determination. The dose ratio between toxicity and therapeutic efficacy is the therapeutic index, and it can be expressed as the ratio of LD 50 to ED 50 . In some or any embodiments, compounds exhibiting a high therapeutic index are preferred. In some embodiments, data obtained from cell culture assays and animal studies are used to formulate the dosage range for human use. In specific embodiments, the dosage of such compounds is within the circulating concentration range that encompasses the ED 50。In some or any embodiments, the dose varies within this range depending on the dosage form employed and the route of administration employed.

[0193] XII. Kit / Product

[0194] There is provided an article of manufacture comprising packaging material; a polymorph of Compound 1 described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition that is effective to modulate the activity of the enzyme GCS or to treat, prevent, or ameliorate one or more symptoms of a GCS-mediated disease, disorder, or condition, within the packaging material; and a label that indicates that the polymorph of Compound 1, the pharmaceutically acceptable salt thereof, or the pharmaceutical composition described herein is for modulating the activity of GCS or for treating, preventing, or ameliorating one or more symptoms of a GCS-mediated disease, disorder, or condition.

[0195] For use in the therapeutic applications described herein, kits and articles of manufacture are also described. In various embodiments, such kits include a carrier, package, or container that is compartmentalized to receive one or more containers (such as vials, tubes, etc.), each of the one or more containers including one of the individual elements for the methods described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. In some embodiments, the containers are formed from a variety of materials (such as glass or plastic).

[0196] In some embodiments, the articles of manufacture described herein contain packaging material. Packaging materials for packaging pharmaceutical products include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, flasks, and any packaging material suitable for the selected formulation and the intended mode of administration and treatment.

[0197] In some embodiments, the containers described herein contain one or more polymorphs of Compound 1 or a pharmaceutically acceptable salt thereof, optionally in a composition or in combination with additional therapeutic agents disclosed herein. The containers optionally have a sterile inlet (e.g., in some embodiments, the container is an intravenous solution bag or a vial with a stopper that can be pierced by a subcutaneous injection needle). Such kits optionally contain a polymorph of Compound 1, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, and an identifying description, label, or instructions related to its use in the methods described herein.

[0198] In some embodiments, the kit will include one or more additional containers, each containing one or more of the various materials (such as reagents, optionally in concentrated form, and / or devices) required for the use of the compounds described herein from a commercial and user perspective. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes; carriers, packages, containers, vials, and / or tube labels listing the contents and / or instructions for use, as well as package inserts with instructions for use. Optionally, a set of instructions is included.

[0199] In some or any embodiments, the label is located on or associated with the container. In some embodiments, the label is located on the container when letters, numbers, or other characters forming the label are attached, molded, or etched into the container itself; the label is associated with the container when the label is present in a receiver or carrier that also holds the container, such as a package insert. In some or any embodiments, the label indicates that the contents are for a specific therapeutic application. In some embodiments, the label indicates the instructions for use of the contents, as in the methods described herein.

[0200] In some or any embodiments, the pharmaceutical composition is present in a packaging or dispensing device containing one or more unit dosage forms, the unit dosage forms containing a crystalline form of compound 1 described herein or a pharmaceutically acceptable salt thereof. In some embodiments, the packaging contains a metal or plastic foil, such as a blister pack. The packaging or dispensing device is optionally accompanied by instructions for administration. In some embodiments, the packaging or dispenser is accompanied by a notice associated with the container, the form of which is specified by a government agency regulating the manufacture, use, or sale of pharmaceuticals, the notice reflecting the approval of the agency for the form of the pharmaceutical for human or veterinary use. In some or any embodiments, such a notice is, for example, a prescription drug label approved by the US Food and Drug Administration, or an approved product insert. In some embodiments, the composition containing a crystalline form of compound 1 described herein or a pharmaceutically acceptable salt thereof is formulated in a compatible pharmaceutical carrier and placed in a suitable container labeled for the treatment of the indicated condition.

[0201] Any combination of the groups described above for the various variables is contemplated herein.

[0202] Examples

[0203] For illustrative purposes, the following examples are included. However, it should be understood that these examples do not limit the invention and are only intended to illustrate the methods of practicing the present disclosure. Those skilled in the art will recognize that the described chemical reactions can be readily adapted to prepare a variety of other compounds and crystal forms of the present disclosure, and alternative methods for preparing the compounds and crystal forms of the present disclosure are considered to be within the scope of the present disclosure. For example, by modifications that are obvious to those skilled in the art, such as by appropriately protecting interfering groups, by utilizing other suitable reagents known in the art in addition to the described reagents, and / or by making routine modifications to the reaction conditions, non-exemplary compounds according to the present disclosure can be successfully synthesized. Alternatively, other reactions disclosed herein or known in the art will be considered suitable for preparing other compounds and crystal forms of the present disclosure.

[0204] For illustrative purposes, the following shows general synthetic schemes for preparing the compounds and crystal forms of the present disclosure and key intermediates. Those skilled in the art will understand that other synthetic schemes can be used to synthesize the compounds and crystal forms. Although specific starting materials and reagents are described and discussed below, other starting materials and reagents can also be easily substituted to provide a variety of derivatives and / or reaction conditions.

[0205] Unless otherwise specified, polymorph A, polymorph B, polymorph C, polymorph D, and polymorph E of Compound 1 described in the specification and examples refer to polymorph A of Compound 1, polymorph B of Compound 1, polymorph C of Compound 1, polymorph D of Compound 1, and polymorph E of Compound 1, respectively.

[0206] The following abbreviations were used in the examples:

[0207]

[0208] Preparation of Compound 1

[0209] The synthesis of Compound 1 is described in detail in Example 173 of WO 2015042397A1, which is reproduced below.

[0210]

[0211]

[0212] Synthesis of Compound A4

[0213] Benzyl chloroformate (50 mL, 50 w.t.% toluene solution, 148 mmol) was added to (R)-2-amino-3-hydroxypropanoic acid (Compound A1) (10.5 g, 100 mmol) in saturated NaHCO 3In an aqueous solution (400 mL). The mixture was vigorously stirred at 20 °C for 4 hours, and the aqueous solution was extracted with diethyl ether (400 mL x 2). The aqueous phase was acidified to pH = 2 with concentrated hydrochloric acid and extracted with ethyl acetate (300 mL x 3). The combined organic phases were dried over Na 2 SO 4 and concentrated to give the crude product compound A2 (20 g, 84% yield) as a white solid. LC-MS (m / z): 240 [M+1] + ; 1 1H-NMR (DMSO-d 6 , 400 MHz) peaks: δ (ppm) 3.653 (m, 2H), 4.051 (m, 1H), 4.884 (m, 1H), 5.038 (s, 2H), 7.303 - 7.373 (m, 6H), 12.658 (s, 1H).

[0214] To a mixture of EDCI·HCl (2.4 g, 12.5 mmol), HOBt (1.7 g, 12.5 mmol), DIPEA (2.7 g, 20 mmol) in DCM (50 mL) was added compound A2 (1 g, 4 mmol) and N,O-dimethylhydroxylamine hydrochloride (1.2 g, 12.5 mmol). The mixture was stirred overnight at room temperature. The mixture was washed with hydrochloric acid solution (1 M, 50 mL x 2), saturated NaHCO 3 aqueous solution (20 mL), brine (20 mL) and dried over Na 2 SO 4 The crude product was purified by silica gel column chromatography (ethyl acetate in petroleum, 30% v / v) to give compound A3 (826 mg, 70% yield) as a colorless liquid. LC-MS (m / z): 283 [M+1] + ; 1 1H-NMR (CDCl 3 , 400 MHz) peaks: δ (ppm) 3.113 (s, 3H), 3.673 (s, 3H), 3.743 (t, J = 4.8 Hz, 2H), 4.766 (m, 1H), 4.959 - 5.044 (m, 2H), 6.046 (d, J = 8.0 Hz, 1H), 7.200 - 7.254 (m 5H).

[0215] At 0 °C, a solution of TBDMS-Cl (800 mg, 5.31 mmol) in THF (10 mL) was added dropwise to a solution of compound A3 (500 mg, 1.77 mmol) and imidazole (602 mg, 8.86 mmol) in THF (20 mL). The mixture was stirred at rt for 2 h and then filtered. The filtrate was washed with 1N HCl (50 mL x 2) and brine (50 mL), and dried over Na 2 SO 4 The crude product was purified by silica gel column chromatography (ethyl acetate in petroleum, 13% v / v) to give compound A4 (526 mg, 75% yield) as a colorless liquid. LC-MS (m / z): 396 [M+1] + ; 1 H-NMR (CDCl 3 , 400 MHz) peaks: δ (ppm) 0.012 (s, 3H), 0.085 (s, 6H), 0.852 (s, 9H), 3.211 (s, 3H), 3.756 (s, 3H), 3.794 - 3.896 (m, 2H), 4.809 (m, 1H), 5.085 (q, J = 11.2 Hz, 2H), 5.662 (d, J = 8.8 Hz, 1H), 7.286 - 7.351 (m 5H).

[0216] Step 1

[0217] At -60 °C under N 2 2, n-BuLi (6.1 mL) was added to a solution of compound 1A (3.73 g, 15.1 mmol) in THF (200 mL), and the mixture was stirred for 0.5 h and then a solution of compound A4 (2 g, 5.06 mmol) in THF (10 mL) was added slowly. The mixture was stirred at -60 °C for 1 h. After adding saturated NH 4 4Cl aqueous solution, the mixture was extracted with ethyl acetate (100 mL x 2), washed with brine (100 mL), and dried over Na 2 SO 4 The crude product was purified by silica gel column chromatography (ethyl acetate in petroleum, 10% v / v) to give compound 1B (1.2 g, 47% yield) as a colorless oil. LC-MS (m / z): 504.2 [M+1] + .

[0218] Step 2

[0219] Compound 1B (714 mg, 1.42 mmol) was dissolved in THF (9 mL) and cooled to -80 °C under a nitrogen atmosphere. Lithium tri-sec-butylborohydride (3 mL of a 1 M THF solution, 18.9 mmol) was added dropwise while maintaining the temperature at -80 °C. After one hour, the reaction mixture was quenched with saturated aqueous NH 4 Cl and extracted with ethyl acetate (50 mL x 2), washed with brine (100 mL), and dried over Na 2 SO 4 . The crude product was purified by silica gel column chromatography (10% ethyl acetate in petroleum ether) to give Compound 1C (350 g, 49% yield) as a colorless oil. LC-MS (m / z): 488.2 [M - 17] + .

[0220] Step 3

[0221] At 0 °C, TBAF (537 mg, 2.06 mmol) was added to a solution of Compound 1C (2.08 g, 4.11 mmol) in THF (40 mL), and the mixture was stirred at room temperature overnight. The mixture was concentrated and then water (50 mL) was added. The mixture was extracted with ethyl acetate (50 mL x 2), washed with brine (100 mL), dried over Na 2 SO 4 , and concentrated. The residue was suspended in a mixture of ethyl acetate and petroleum ether (10% v / v). Filtration gave Compound 1D (1 g, 65% yield) as a white solid. LC-MS (m / z): 374.1 [M + 1] + .

[0222] Step 4

[0223] To a solution of Compound 1D (0.9 g, 2.3 mmol) in THF (60 mL) was added Et 3 N (700 mg, 6.9 mmol), and the mixture was cooled to -15 °C. MsCl (467 mg, 4.1 mmol) was slowly added to the mixture. The mixture was stirred at -15 °C for about half an hour and then diluted with water. The mixture was extracted with ethyl acetate (50 mL x 2), washed with brine (50 mL), dried over Na 2 SO 4 , and concentrated to give Compound 1E (1.12 g, crude product), which was used in the next step without further purification. LC-MS (m / z): 452.1 [M + 1] + .

[0224] Step 5

[0225] To a solution of compound 1E (1.12 g, 2.83 mmol) in THF (60 mL) was added pyrrolidine (2 g, 28 mmol). The reaction mixture was warmed to room temperature and then heated at 50 °C overnight. The crude product was purified by silica gel column chromatography (methanol in dichloromethane, 5% v / v) to give compound 1F (0.5 g, 50% yield). LC-MS (m / z): 445 [M+1] + 。

[0226] Step 6

[0227] To a solution of compound 1F (520 mg, 1.17 mmol) in ethanol (12 mL) and water (2 mL) was added LiOH H 2 O (197 mg, 4.68 mmol). The mixture was stirred at 80 °C overnight, diluted with water, extracted with DCM (50 mL x 2), washed with brine (50 mL), dried over Na 2 SO 4 and concentrated to give compound 1G (360 mg, crude), which was used in the next step without further purification. LC-MS (m / z): 311.1 [M+1] + 。

[0228] Step 7

[0229] A mixture of compound 1H (137 mg, 0.59 mmol), HATU (445 mg, 1.17 mmol), DMF (4 mL) and compound 1G (218 mg, 0.70 mmol) in DCM (16 mL) was stirred at room temperature overnight. The reaction mixture was treated with water (20 mL), extracted with DCM (50 mL x 2), washed with brine (50 mL), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by preparative HPLC to give compound 1I. LC-MS (ESI) m / z: 527 [M+H] + ; 1H-NMR(CD3OD, 400 MHz): δ(ppm) 0.54 - 0.79 (m, 4H), 2.04 - 2.22 (m, 5H), 3.23 - 3.26 (m, 1H), 3.58 - 3.82 (m, 6H), 4.70 - 4.73 (m, 1H), 4.99 (d, J = 2.8 Hz, 1H), 7.30 (d, J = 8.4 Hz, 1H), 7.37 - 7.41 (m, 1H), 7.50 (d, J = 2.0 Hz, 1H), 7.57 - 7.59 (m, 1H), 7.88 - 7.93 (m, 3H), 7.99 (d, J = 1.2 Hz, 1H), 8.29 (s, 1H).

[0230] Step 8

[0231] A mixture of compound 1I (40.8 mg, 0.08 mmol), hydroxylamine hydrochloride (76 mg, 1.1 mmol) and methanol (9 mL) was stirred overnight at 50 °C. The mixture was then purified by preparative HPLC to afford compound 1. LC-MS (ESI) m / z: 542.1 [M + H] + ; 1 H-NMR (MeOD, 400 MHz) δ(ppm) 0.69 - 0.84 (m, 4H), 2.02 - 2.22 (m, 4H), 3.21 - 3.29 (m, 1H), 3.62 - 3.80 (m, 5H), 4.70 - 4.73 (m, 1H), 5.01 (d, J = 2.8 Hz, 1H), 7.32 - 7.34 (m, 1H), 7.39 - 7.43 (m, 1H), 7.45 - 7.49 (m, 1H), 7.51 - 7.53 (m, 1H), 7.63 - 7.85 (m, 3H), 7.88 - 7.90 (m, 1H).

[0232] General procedure

[0233] Generally, XRPD analysis, DSC analysis, TGA analysis, PLM analysis, Karl Fischer (KF) analysis and 1 H NMR analysis were used in the following examples. The instruments and conditions for each analysis are listed below.

[0234]

[0235]

[0236] Example 1: Preparation, characterization and evaluation of crystalline form A of compound 1

[0237] Preparation of Polymorph A of Compound 1

[0238] (1) Equilibrium experiments at 25 °C

[0239] Approximately 50 mg of the crystalline material was equilibrated in an appropriate amount of different solvents (as shown in Table 1 below) at 25 °C using a magnetic stirrer plate for 2 weeks. The resulting suspension was filtered. The solid portion (wet filter cake) was studied by XRPD. Each of the following solvents used in Table 1 provided polymorph A of Compound 1.

[0240] Table 1 Equilibration at 25 °C with different solvents for 2 weeks

[0241] Example No. Solvent 1.1 Water 1.2 Ethanol 1.3 2 - Propanol 1.4 Acetone 1.5 Methyl Ethyl Ketone 1.6 Acetonitrile 1.7 Ethyl Acetate 1.8 Isopropyl Acetate 1.9 1,4 - Dioxane 1.10 Tetrahydrofuran 1.11 Dichloromethane 1.12 Methanol / Water (40:60, v / v) a.w. 0.8 1.13 Acetone / Water (50:50, v / v) a.w. 0.9 1.14 Acetonitrile / Water (90:10, v / v) a.w. 0.9

[0242] (2) Equilibrium experiments at 50 °C

[0243] Approximately 50 mg of the crystalline material was equilibrated in an appropriate amount of different solvents (as shown in Table 2 below) at 50 °C using a magnetic stirrer plate for 1 week. The resulting suspension was filtered. The solid portion (wet filter cake) was studied by XRPD. Each of the following solvents used in Table 2 provided polymorph A of Compound 1.

[0244] Table 2 Equilibration at 50 °C with different solvents for 1 week

[0245] Example No. Solvent 1.15 Water 1.16 Ethanol 1.17 2 - Propanol 1.18 Acetone 1.19 Methyl Ethyl Ketone 1.20 Acetonitrile 1.21 Ethyl Acetate 1.22 Isopropyl Acetate 1.23 1,4 - Dioxane 1.24 Tetrahydrofuran 1.25 Toluene 1.26 Methanol / Water (40:60, v / v) a.w. 0.8 1.27 Acetone / Water (50:50, v / v) a.w. 0.9 1.28 Acetonitrile / Water (90:10, v / v) a.w. 0.9

[0246] (3) Anti-solvent experiments

[0247] 30 mg - 50 mg of the crystalline material was dissolved in different solvents (as shown in Table 3 below). The resulting solution was filtered through a 0.45 μm nylon filter to obtain a clear solution. Each anti-solvent was slowly added to the clear solution. The precipitate was collected by filtration. The solid portion (wet filter cake) was studied by XRPD. Each solvent / anti-solvent system used in Table 3 provided polymorph A of Compound 1.

[0248] Table 3 Precipitation by addition of anti-solvent

[0249]

[0250]

[0251] Note: "V" refers to the volume ratio of anti-solvent to solvent.

[0252] (4) Crystallization by slow evaporation at room temperature

[0253] The crystalline material was dissolved in an appropriate amount of acetone / acetonitrile (1:1, v / v). The resulting solution was filtered through a 0.45 μm nylon filter. The resulting clear solution was slowly evaporated under ambient conditions. The solid residue was studied by XRPD and was identified as polymorph A of Compound 1.

[0254] (5) Rapid cooling

[0255] At 50 °C, 30 mg - 50 mg of the crystalline material was dissolved in the minimum amount of the selected solvent (as shown in Table 4 below). The resulting solution was placed in an ice bath and stirred. The precipitate was collected by filtration. The solid portion (wet filter cake) was studied by XRPD. Each of the following solvents provided polymorph A of Compound 1: ethanol, acetone, methyl ethyl ketone, and 1,4 - dioxane. Using this method, tetrahydrofuran, dichloromethane, acetonitrile / tetrahydrofuran (1:1, v / v), and ethyl acetate / tetrahydrofuran (1:1, v / v) did not provide polymorph A of Compound 1. When no precipitate was obtained, the solution was placed in a -20 °C refrigerator for crystallization.

[0256] Table 4 Crystallization from a hot saturated solution by rapid cooling

[0257]

[0258]

[0259] Characterization of Polymorph A of Compound 1

[0260] (1) XRPD data

[0261] The XRPD pattern of polymorph A of Compound 1 is as shown in Figure 1 . The peaks in the XRPD pattern and their relative intensities are as shown in Table 5 below.

[0262] Table 5 Peaks in the XRPD pattern of polymorph A of Compound 1

[0263]

[0264]

[0265] (2) TGA and DSC data

[0266] The TGA curve and DSC curve of polymorph A of Compound 1 are shown in Figure 2 and Figure 3 respectively. As shown in Figure 2 , a weight loss of approximately 0.6% was observed between approximately 24 °C and approximately 200 °C, indicating that polymorph A of Compound 1 is a non - hydrated polymorph. As shown in Figure 3 , the melting point of polymorph A of Compound 1 is 206.9 °C (97 J / g), and decomposition occurs upon melting.

[0267] (3) 1 1H NMR data

[0268] For polymorph A of Compound 11 The \(^1\)H NMR spectrum is as Figure 4 shown in. As Figure 4 shown, no residual solvent was detected by 1 \(^1\)H NMR.

[0269] (4) KF data

[0270] KF analysis showed that the crystalline form A of Compound 1 contains approximately 0.12 wt% water.

[0271] Evaluation of Polymorph A of Compound 1

[0272] (1) Stability

[0273] The crystalline form A of Compound 1 was chemically and physically stable for one week in an open container at 25 °C / 92% RH, chemically and physically stable for one week in an open container at 40 °C / 75% RH, and chemically and physically stable for one week in a sealed container at 60 °C (see Table 20 in Example 12 below). Given the above stability evaluation results, the crystalline form A of Compound 1 is highly stable.

[0274] (2) Hygroscopicity

[0275] The hygroscopicity of the crystalline form A of Compound 1 was evaluated by DVS test. The DVS plot of the crystalline form A of Compound 1 is shown in Figure 26 . According to the DVS test results, in the DVS study, when the relative humidity increased from 40% Rh to 90% RH at 25 °C, the water absorption rate increased from 0.06% to 0.39%, indicating that the crystalline form A of Compound 1 is slightly hygroscopic. As shown by the XRPD patterns of the crystalline form A of Compound 1 before and after the DVS test in Figure 27 , no crystalline form change was observed after the DVS test. All data indicate that the crystalline form A of Compound 1 is a highly crystalline anhydrate.

[0276] Example 2: Preparation and Characterization of the Crystalline Form B of Compound 1

[0277] Preparation of Polymorph B of Compound 1

[0278] (1) Equilibrium experiment at 25 °C

[0279] The crystalline form A of Compound 1 was used in this method. Approximately 50 mg of the crystalline material was equilibrated in an appropriate amount of methanol at 25 °C for 2 weeks using a magnetic stirrer. The resulting suspension was filtered. The solid portion (wet filter cake) was characterized by XRPD and identified as the crystalline form B of Compound 1.

[0280] (2) Equilibrium experiment at 50 °C

[0281] Polymorph A of Compound 1 is used in this method. About 50 mg of the crystalline material is equilibrated in an appropriate amount of methanol at 50 °C using a magnetic stirrer for 1 week. The obtained suspension is filtered. The solid part (wet filter cake) is characterized by XRPD and identified as polymorph B of Compound 1.

[0282] (3) Scale-up experiment of polymorph B of Compound 1

[0283] Polymorph A of Compound 1 is used in this method. At 25 °C, about 500 mg of the crystalline material is equilibrated in 10 mL of methanol using a magnetic stirrer for about 5 days. The obtained solid is recovered by filtration. The solid part (wet filter cake) is dried under ambient conditions for about 3 hours. The obtained dry solid is characterized by XRPD and identified as polymorph B of Compound 1. About 447 mg of polymorph B of Compound 1 as a white solid with a yield of 89% is obtained.

[0284] Characterization of Polymorph B of Compound 1

[0285] (1) XRPD data

[0286] The XRPD pattern of polymorph B of Compound 1 is as shown in Figure 5 . The peaks in the XRPD pattern and their relative intensities are as shown in Table 6 below.

[0287] Table 6 Peaks in the XRPD pattern of polymorph B of Compound 1

[0288]

[0289]

[0290] (2) TGA and DSC data

[0291] The TGA curve and DSC curve of polymorph B of Compound 1 are shown in Figure 6 and Figure 7 respectively. As shown in Figure 6 , a weight loss of about 0.4% is observed between about 33 °C and 200 °C. As shown in Figure 7 , polymorph B of Compound 1 has multiple thermal events at initial temperatures of 178.3 °C (0.7 J / g), 179.7 °C (0.9 J / g), 196.6 °C (74 J / g), 198.7 °C (10 J / g), and 203.7 °C (22 J / g), respectively.

[0292] (3) 1 1H NMR data

[0293] The 1 1H NMR spectrum of polymorph B of Compound 1 is as shown in Figure 8 . As shown in Figure 8As shown, through 1 H-NMR detected no residual methanol. Equilibrating the wet filter cake obtained from methanol at 25 °C showed the same XRPD pattern as polymorph B of Compound 1. Without air drying, through 1 H-NMR detected approximately 38.8 wt% methanol (about 10 equivalents) in polymorph B of the wet filter cake of Compound 1. Based on the results, polymorph B of Compound 1 may be a highly crystalline methanol solvate.

[0294] Example 3: Preparation, Characterization, and Evaluation of Polymorph C of Compound 1

[0295] Preparation of Polymorph C of Compound 1

[0296] Polymorph A of Compound 1 was used in this method. Polymorph C of Compound 1 was prepared from methanol / dichloromethane (1:1, v / v) by slow evaporation experiments. Specifically, approximately 30 mg of polymorph A of Compound 1 was dissolved in an appropriate amount of methanol / dichloromethane (1:1, v / v). The obtained solution was filtered through a 0.45 μm nylon filter. The obtained clear solution was slowly evaporated under ambient conditions. The solid residue was studied by XRPD and identified as polymorph C of Compound 1.

[0297] In addition, a scale-up experiment of polymorph C of Compound 1 was carried out. Specifically, approximately 120 mg of polymorph A of Compound 1 was dissolved in 8 mL of methanol / dichloromethane (1 / 1, v / v). The obtained solution was filtered through a 0.45 μm nylon filter to obtain a clear solution. Then the clear solution was rapidly evaporated under a nitrogen stream. After drying under ambient conditions for about 1 day, approximately 106 mg of polymorph C of Compound 1 as a white solid with a yield of about 87% was successfully obtained. The obtained dried solid was characterized by XRPD and identified as polymorph C of Compound 1.

[0298] Characterization of Polymorph C of Compound 1

[0299] (1) XRPD Data

[0300] The XRPD pattern of polymorph C of Compound 1 is as shown in Figure 9 . The peaks in the XRPD pattern and their relative intensities are shown in Table 7 below.

[0301] Table 7 Peaks in the XRPD Pattern of Polymorph C of Compound 1

[0302]

[0303]

[0304] (2) TGA and DSC Data

[0305] The TGA curve and DSC curve of crystalline form C of Compound 1 are shown in Figure 10 and Figure 11 respectively. As shown in Figure 10 , a weight loss of approximately 2.2% was observed at about 150 °C. No distinct melting peak was observed in the DSC curve ( Figure 11 ).

[0306] (3) 1 1H NMR data

[0307] The 1 1H NMR spectrum of crystalline form C of Compound 1 is shown in Figure 12 . As shown in Figure 12 , no residual solvent was detected by 1 1H-NMR.

[0308] (4) KF data

[0309] KF analysis showed that crystalline form C of Compound 1 contains approximately 3.6 wt% water (1.1 equivalents), indicating that crystalline form C of Compound 1 is a hydrate containing approximately 3.6 wt% water (1.1 equivalents).

[0310] Evaluation of Polymorph C of Compound 1

[0311] See Example 6: Details of the water activity study are given below.

[0312] Example 4: Preparation, Characterization and Evaluation of Crystalline Form D of Compound 1

[0313] Preparation of Polymorph D of Compound 1

[0314] Approximately 150 mg of crystalline form B of Compound 1 in 20 batches was heated to 190 °C by TGA and then cooled to ambient temperature. Approximately 110 mg of a light white solid (approx. 73% yield) was obtained. The dried solid obtained was characterized by XRPD and identified as crystalline form D of Compound 1.

[0315] Characterization of Polymorph D of Compound 1

[0316] (1) XRPD data

[0317] The XRPD pattern of crystalline form D of Compound 1 is shown in Figure 13 . The peaks in the XRPD pattern and their relative intensities are shown in Table 8 below.

[0318] Table 8 Peaks in the XRPD Pattern of Crystalline Form D of Compound 1

[0319]

[0320]

[0321] (2) TGA and DSC data

[0322] The TGA curve and DSC curve of polymorph D of Compound 1 are shown in Figure 14 and Figure 15 respectively. As Figure 14 shown, a weight loss of approximately 0.4% was observed between approximately 33 °C and approximately 170 °C. As Figure 15 shown, polymorph D of Compound 1 showed an exothermic peak starting at 88.7 °C (8 J / g). Thereafter, a small endothermic peak starting at 172.8 °C (2 J / g) was observed. Then, a sharp melting peak with an enthalpy of approximately 90 J / g starting at 207.4 °C was observed. The data indicate that polymorph D of Compound 1 may be a highly crystalline anhydrate.

[0323] (3) 1 1H NMR data

[0324] The 1 1H NMR spectrum of polymorph D of Compound 1 is shown in Figure 16 respectively. As Figure 16 shown, no residual solvent was detected by 1 1H-NMR.

[0325] Evaluation of Polymorph D of Compound 1

[0326] See Example 7: Details of the competitive equilibrium experiment are given below.

[0327] Example 5: Preparation, Characterization, and Evaluation of Polymorph E of Compound 1

[0328] Preparation of Polymorph E of Compound 1

[0329] The crude product (18.5 g, 32.0 mmol) was purified by an SFC column: Phenomenex - cellulose - 2 (250 mm * 30 mm, 10 um); mobile phase: [0.1% NH 3 H 2 O MeOH]; B%: 60% - 60%, 3.9 minutes; 1770 minutes) to obtain P1. Then P1 was concentrated to obtain Residue 1. EtOH (100 mL) was added to Residue 1, and the resulting solution was concentrated. This post-treatment was repeated three times to obtain Residue 2. H 2 O (100 mL) was added to Residue 2, and the resulting solution was lyophilized. Polymorph E of Compound 1 (10.3 g, 18.5 mmol) was obtained.

[0330] Characterization of Polymorph E of Compound 1

[0331] The XRPD pattern of crystalline form E of Compound 1 is as Figure 17 shown. The peaks in the XRPD pattern and their relative intensities are shown in Table 9 below.

[0332] Table 9 Peaks in the XRPD Pattern of Crystalline Form E of Compound 1

[0333]

[0334]

[0335] Evaluation of Polymorph E of Compound 1

[0336] See Example 7: Details of the competitive equilibrium experiment are given below.

[0337] Example 6: Water Activity Study

[0338] The water activity experiment was carried out at 25 °C or 50 °C to determine the critical water activity between the anhydrous form and the hydrate. At 25 °C or 50 °C, saturated solutions were prepared with an ethanol / water mixture at 8 different water activities using the anhydrous crystalline form A of Compound 1. 4 mg of crystalline form A of Compound 1 and 4 mg of crystalline form C of Compound 1 were added to the saturated solutions. The mixtures were stirred at 25 °C or 50 °C for at least 2 days. The solid portion obtained (wet filter cake) was studied by XRPD.

[0339] Tables 10 and 11 show the results of the water activity study in different solvents at 25 °C and 50 °C, respectively. The water activity of the binary solvent system was calculated based on the UNIFAC method (UNIQUAC group activity coefficients). As shown in Tables 10 and 11, crystalline form A of Compound 1 was obtained in all experiments, indicating that crystalline form C of Compound 1 is a metastable hydrate and crystalline form A of Compound 1 is stable over a wide range of water activities.

[0340] Table 10 Water Activity Study Conducted at 25 °C Using Crystalline Forms A and C of Compound 1

[0341]

[0342] Table 11 Water Activity Study Conducted at 50 °C Using Crystalline Forms A and C of Compound 1

[0343]

[0344]

[0345] Example 7: Competitive Equilibrium Experiment

[0346] A suspension competition experiment was conducted to determine the thermodynamic relationship among polymorphic forms A, D, and E of Compound 1. At 5 °C, 25 °C, and 50 °C respectively, saturated solutions were prepared in solvents using anhydrous polymorphic form A of Compound 1. 4 mg of polymorphic form A of Compound 1, 4 mg of polymorphic form D of Compound 1, and 4 mg of polymorphic form E of Compound 1 were added to the saturated solutions. The mixtures were stirred at 5 °C, 25 °C, and 50 °C for at least 3 days. The solid portion (wet filter cake) obtained was studied by XRPD.

[0347] Table 12 below shows the results of the competitive equilibrium experiments in different solvents at 5 °C, 25 °C, and 50 °C. As shown in Table 12, polymorphic form A of Compound 1 is the only product in different solvents at 5 °C, 25 °C, and 50 °C, indicating that polymorphic form A of Compound 1 is the most stable polymorphic form.

[0348] Overall, in these examples, polymorphic form A of Compound 1 was confirmed to be the most stable anhydrous form and the best polymorph. Therefore, polymorphic form A of Compound 1 is recommended for further development.

[0349] Table 12 Results of the competitive equilibrium experiments

[0350]

[0351] Example 8: Salt form screening

[0352] The free form polymorphic form A of Compound 1 was used as the starting material for salt form screening and selection studies. Approximately 5 mg of the free form polymorphic form A of Compound 1 was weighed into a 2 mL glass bottle, and 20 μL aliquots of each solvent were added to obtain a clear solution. The maximum volume of each solvent added was 1 mL. The approximate solubility of the starting material was determined by visual observation at 25 °C. The approximate solubility of the starting material at 25 °C is shown in Table 13 below.

[0353] Table 13 Approximate solubility at 25 °C

[0354] Example No. Solvent Solubility (mg / mL, 25°C) 8.1 Ethanol 5-5.5 8.2 Acetone 16.7-25 8.3 Ethyl Acetate <5 8.4 Acetonitrile <5 8.5 Water <5

[0355] Based on the calculated pKa of 7.85 (calculated by Marvin Sketch 5.6), 5 counterions were selected as salt formers, as shown in Table 14 below. Acetone and ethanol / water (95:5, v / v) were used as screening solvents. A total of approximately 19 salt form screening experiments were conducted.

[0356] Table 14 Counterions for salt form screening

[0357] Counterion pKa(s) M.W. Grade Hydrochloric Acid -6 36.46 I L - Malic Acid 3.4 134.09 I L - Tartaric Acid 3.02 150.09 I Citric Acid 3.12 192.13 I Fumaric Acid 3.03 116.08 I

[0358] Approximately 50 mg of crystalline form A of the free form of compound 1 was added to a suitable solvent, and 1.05 equivalents of a counterion were added. Before addition, the liquid counterion was first diluted with 10 times the solvent. The resulting mixture was stirred at 50 °C for 1 hour and then stirred at 25 °C for at least 36 hours.

[0359] For those clear solutions, the solution was cooled to 5 °C and maintained at 5 °C for at least 24 hours. If no precipitate was found, an anti-solvent (ethyl acetate and heptane) was added to the clear solution. The resulting sample was stirred at 5 °C for about 7 days.

[0360] The resulting suspension was taken out and centrifuged. The resulting solid was dried under vacuum at 30 - 40 °C for at least 12 hours and analyzed by XRPD. The results are summarized in Tables 15 and 16 below. The hit salt forms with high or medium crystallinity were further characterized.

[0361] Table 15 Salt form screening results (suspension crystallization)

[0362]

[0363] Note: "+": hit salt form

[0364] "-": free form, counterion or physical mixture

[0365] Table 16 Salt form screening results (anti-solvent addition)

[0366]

[0367] Note: "+": hit salt form

[0368] "-": free form, counterion or physical mixture

[0369] "AF": amorphous form

[0370] " / / ": not performed

[0371] Example 9: Preparation of salt form candidates

[0372] From salt form screening, four salt forms and their polymorphs were identified, including Form A of the L-malate of Compound 1, Form A of the L-tartrate of Compound 1, Form B of the L-tartrate of Compound 1, Form A of the fumarate of Compound 1, Form B of the fumarate of Compound 1, and Form A of the hydrochloride of Compound 1. Among these salts, Form A of the L-malate of Compound 1 and Form A of the L-tartrate of Compound 1 showed good physicochemical properties, including high crystallinity, high melting point, and good stoichiometry; Form B of the L-tartrate of Compound 1 and Form A of the fumarate of Compound 1 were semi-salts with low crystallinity; Form B of the fumarate of Compound 1 had a high melting point and reasonable stoichiometry, but medium crystallinity; Form A of the hydrochloride of Compound 1 was an amorphous form. Therefore, Form A of the L-malate of Compound 1 and Form A of the L-tartrate of Compound 1 were selected as candidate salt forms and scaled up for comprehensive evaluation.

[0373] Form A of the L-malate of Compound 1 and Form A of the L-tartrate of Compound 1 were scaled up to approximately 500 mg and compared and evaluated with the free form Form A of Compound 1. The procedures for preparing Form A of the L-malate of Compound 1 and Form A of the L-tartrate of Compound 1 are shown in Table 17 below.

[0374] Table 17 Preparation of Salt Form Candidates

[0375]

[0376]

[0377] Example 10: Characterization of Salt Candidate Form A of the L-malate of Compound 1

[0378] (1) XRPD Data

[0379] The XRPD pattern of Form A of the L-malate of Compound 1 is shown in Figure 18 . The peaks in the XRPD pattern and their relative intensities are shown in Table 18 below. The XRPD pattern of Form A of the L-malate of Compound 1 indicates that Form A of the L-malate of Compound 1 is a highly crystalline anhydrate.

[0380] Table 18 Peaks in the XRPD Pattern of Form A of the L-malate of Compound 1

[0381]

[0382]

[0383]

[0384] (2) TGA and DSC data

[0385] The TGA curve and DSC curve of polymorph A of L-malate of Compound 1 are shown in Figure 19 and Figure 20 respectively. As Figure 19 shown, a weight loss of about 0.8% was observed between about 31 °C and about 156 °C. As Figure 20 shown, polymorph A of L-malate of Compound 1 has an onset melting peak at about 160.3 °C and an enthalpy of about 68 J / g.

[0386] (3) 1 1H NMR data

[0387] The 1 1H NMR spectrum of polymorph A of L-malate of Compound 1 is shown in Figure 21 indicating that no residual solvent was detected.

[0388] (4) KF data

[0389] KF analysis shows that polymorph A of L-malate of Compound 1 contains about 0.3 wt% water.

[0390] Example 11: Characterization of Salt Candidate Polymorph A of L-tartrate of Compound 1

[0391] (1) XRPD data

[0392] The XRPD pattern of polymorph A of L-tartrate of Compound 1 is shown in Figure 22 respectively. The peaks in the XRPD pattern and their relative intensities are shown in Table 19 below. The XRPD pattern of polymorph A of L-tartrate of Compound 1 indicates that polymorph A of L-tartrate of Compound 1 is a highly crystalline anhydrate.

[0393] Table 19 Peaks in the XRPD Pattern of Polymorph A of L-tartrate of Compound 1

[0394]

[0395]

[0396]

[0397] (2) TGA and DSC data

[0398] The TGA curve and DSC curve of polymorph A of L-tartrate of Compound 1 are shown in Figure 23 and Figure 24 respectively. As Figure 23As shown, a weight loss of about 0.8% was observed between about 32 °C and about 185 °C. As Figure 24 shown in

[0399] (3) 1 1H NMR data

[0400] The 1H NMR spectrum of Form A of the L-tartrate salt of Compound 1 1 is shown in Figure 25 .

[0401] (4)KF data

[0402] KF analysis indicated that Form A of the L-tartrate salt of Compound 1 contains about 0.6 wt% water.

[0403] Example 12: Evaluation of Salt Candidates

[0404] Form A of the L-malate salt of Compound 1 and Form A of the L-tartrate salt of Compound 1 were successfully scaled up. The scaled-up batches were the same polymorph as the screening samples. These two candidate salt forms were compared with the free form polymorph of Compound 1 to evaluate their stability, physicochemical properties, solubility, and hygroscopicity.

[0405] Stability

[0406] The purity and appearance of the free form polymorph of Compound 1, Form A of the L-malate salt of Compound 1, and Form A of the L-tartrate salt of Compound 1 are shown in Table 20 below. As shown in Table 20, the high chemical purity of the free form polymorph of Compound 1, Form A of the L-malate salt of Compound 1, and Form A of the L-tartrate salt of Compound 1 is 99.8%; the free form polymorph of Compound 1, Form A of the L-malate salt of Compound 1, and Form A of the L-tartrate salt of Compound 1 are chemically and physically stable for one week in open containers at 25 °C / 92% RH, in open containers at 40 °C / 75% RH, and in sealed containers at 60 °C.

[0407] Table 20 Stability: Purity and Appearance

[0408]

[0409] Notes:

[0410] "NC": No color change

[0411] "CL": Color

[0412] Crystallinity and Physicochemical Properties

[0413] The chemical and physicochemical properties of crystalline form A of the free form of Compound 1, crystalline form A of the L-malate salt of Compound 1, and crystalline form A of the L-tartrate salt of Compound 1 are shown in Table 21 below.

[0414] Solubility

[0415] As shown in Table 21 below, solubility was tested for 2 hours and 24 hours at 37 °C in 4 different pH buffers and 3 biorelevant media. As a weak base, Compound 1 shows typical pH-dependent solubility, with high solubility at low pH and low solubility at neutral pH. Except in water and the pH 4.5 buffer, the solubility of crystalline form A of the free form of Compound 1 and the two candidate salts is similar in most media. The free base and salts show good solubility in the pH 1.2 buffer, SGF, and FeSSIF-V1. The higher solubility of the two salt candidates in water is due to the lower pH caused by the acidic counterions. In the pH 4.5 buffer, crystalline form A of the L-malate salt of Compound 1 and crystalline form A of the L-tartrate salt of Compound 1 show solubilities approximately 3-fold and 2-fold higher, respectively, than the solubility of crystalline form A of the free form of Compound 1. However, this solubility increase is not due to the salts, but because the two salts disproportionate to form an amorphous free form in the buffer. Amorphous forms generally have higher solubility than crystalline forms. Crystalline form A of the free form of Compound 1 and the salts show higher solubility (>2 mg / mL) in FeSSIF-V1 than in buffers of similar pH, indicating that surfactants may facilitate solubility enhancement.

[0416] Morphological Properties

[0417] Hygroscopicity: The hygroscopicity of crystalline form A of the free form of Compound 1 and the salt candidates was evaluated by dynamic vapor sorption (DVS) testing. Crystalline form A of the free form of Compound 1 is slightly hygroscopic, with a water uptake increase of approximately 0.33% from 40% Rh to 90% RH at 25 °C, and no morphological changes were observed after the DVS test. Crystalline form A of the L-malate salt of Compound 1 is slightly hygroscopic, with a water uptake increase of approximately 0.59% from 40% Rh to 90% RH at 25 °C, and no morphological changes were observed after the DVS test. Crystalline form A of the L-tartrate salt of Compound 1 is slightly hygroscopic, with a water uptake increase of approximately 0.39% from 40% Rh to 90% RH at 25 °C, and no morphological changes were observed after the DVS test.

[0418] Table 21 Chemical and physicochemical properties of crystalline form A of the free form of Compound 1, crystalline form A of the L-malate salt of Compound 1, and crystalline form A of the L-tartrate salt of Compound 1

[0419]

[0420]

[0421]

[0422] In summary, for oral applications, the free form polymorph A of compound 1, the polymorph A of the L-malate salt of compound 1, and the polymorph A of the L-tartrate salt of compound 1 showed good crystallinity, high melting points, and reasonable stoichiometry. Except in buffer at pH 4.5, they also exhibited satisfactory stability, slight hygroscopicity, and reasonable solubility. PK studies showed that the free form and the two salts had similar PK.

[0423] Example 13: Biological Activity Study

[0424] In this study, the inventors developed a reliable method to monitor GCS catalytic activity using the fluorescent substrate C6-NBD-ceramide. The reaction product C6-NBD-glucosylceramide could then be detected and quantified by LC-MS / MS. The present inventors used this method to test and validate the effects of various GCS inhibitors on GCS enzyme activity from different species.

[0425] The MDCK, K-562, and L6 cell lines all endogenously express the GCS enzyme and were used to test the effects of various GCS inhibitors on the enzyme from different species. GCS enzyme activity was detected by using the fluorescent substrate C6-NBD-ceramide and monitoring the formation of the fluorescent reaction product C6-NBD-glucosylceramide. First, the inventors used cell lysates from the canine kidney cell line (MDCK, ATCC catalog number CCL-34 TM ) as a source of GCS and tested the inhibitory activity of several control compounds and test compounds. Then, the inventors used the human leukemia K-562 cell line (ATCC catalog number CRL-1458 TM ) to test the effects of these GCS inhibitors on intact live cells. Finally, cell lysates from the rat skeletal muscle cell line L6 (ATCC catalog number CCL-243 TM ) were used to test the effects of these GCS inhibitors on rodent GCS enzyme.

[0426] The information on the test compounds and control compounds is listed below.

[0427] Table 22 Test Compounds

[0428]

[0429] Table 23 Control Compounds

[0430]

[0431] GCS activity test by enzymatic assay

[0432] The GCS activity was evaluated using a GCS enzyme assay derived from MDCK or L6 cell lysates.

[0433] In the presence of a protease inhibitor mixture, MDCK or L6 cell lysates were prepared using M-PER. Each 100 mm culture dish (100% confluent) was lysed with 250 μl of M-PER containing protease inhibitors. The protein concentration was determined using the Pierce BCA Protein Assay Kit.

[0434] Sixty micrograms of MDCK lysate was incubated with various concentrations of the compound (0.00001 μM - 1 μM) at 37 °C for 1 hour in 100 mM Tris buffer (pH 7.5) with a final reaction volume of 100 μl, which contained 10 mM MgCl 2 , 1 mM dithiothreitol, 1 mM EGTA, 2 mM NAD, 100 μM UDP-glucose, 10 μM C6-NBD-ceramide, and 35 μM dioleoyl phosphatidylcholine / 5 μM sulfatide. The final concentration of DMSO was 0.1% in both the compound-treated and DMSO-only treated samples. Each individual reaction was terminated with 11 volumes of methanol:acetonitrile (6:5; containing 200 ng / ml tolbutamide as an internal control). After thorough mixing, the reaction mixture was centrifuged at 2000 rpm for 10 minutes, and the supernatant was injected into LC-MS / MS for analysis.

[0435] For the assay using L6 cell lysates, different amounts of L6 cell lysates were used.

[0436] GCS activity test by cell-based assay

[0437] The GCS activity was evaluated using a GCS cell-based assay with K-562 cells.

[0438] K-562 cells were seeded into a 12-well plate (3×10 5Cells / well, 1 ml), and incubated at 37 °C for 24 hours. These cells were incubated with different concentrations of the compound (0.00001 μM - 1 μM) for 4 hours. Then, 100 μl of [RPMI-1640 + 5% FBS + 11% BSA + 110 μM C6-NBD-ceramide] containing the compound or DMSO was added to the cells, mixed well, and incubated at 37 °C for 0.5 hour. After incubation, the cells were centrifuged, washed twice with ice-cold DPBS (pH 7.4), and resuspended in 50 μl of cold DPBS + 1% Triton X-100. The mixture was sonicated for 15 minutes. Small aliquots (2.5 μl) were used to determine the protein concentration. Then, 11 volumes of methanol:acetonitrile (6:5; containing 200 ng / ml tolbutamide as an internal control) were added to each sample. After mixing well, the samples were centrifuged at 2000 rpm for 10 minutes, and the resulting supernatant was injected into LC-MS / MS for analysis.

[0439] Quantitative analysis of C6-NBD-glucosylceramide was performed on a Shimadzu ultra-fast liquid chromatography and API 4000 mass spectrometer. The Analyst 1.5 software package was used to control the LC-MS / MS system and for data acquisition and processing.

[0440] C6-NBD-glucosylceramide readings were first normalized by dividing the peak area of C6-NBD-glucosylceramide by the peak area of the internal standard, and then the amount was calculated according to the standard curve generated in the same assay run. For cell-based assay results, the amount of C6-NBD-glucosylceramide was also normalized by the protein concentration of the cell samples.

[0441] Using GraphPad Prism software (GraphPad Software, Inc., San Diego, CA), the inhibition percentage of C6-NBD-glucosylceramide accumulation relative to the DMSO control was used to generate an IC 50 value from a sigmoidal dose-response (variable slope) curve.

[0442] Test of the effect of GCS inhibitors using MDCK cell lysates

[0443] The effect of GCS inhibitors was tested using MDCK cell lysates as a source of GCS enzyme. The compounds eliglustat and venglustat were used as controls. Polymorph A of compound 1 was tested. The results of the GCS inhibitor effect test performed with MDCK cell lysates are shown in Figure 32 A (eliglustat), Figure 32 B (venglustat) and Figure 32In polymorph A of Compound 1 and is also summarized in Table 24 below. The results showed that, compared with the control, test compound polymorph A of Compound 1 had better inhibitory effect on GCS activity.

[0444] Table 24 Summary of the effects of GCS inhibitors on MDCK cell lysates

[0445] Compound <![CDATA[IC 50 (nM)]]> Elubrixostat 52.20 Venglustat 72.15 Polymorph A of Compound 1 7.66

[0446] Test of the effect of GCS inhibitors using live K-562 cells

[0447] The effects of GCS inhibitors were tested using live K-562 cells as the source of GCS enzyme. Compounds eliglustat and venglustat were used as controls. Polymorph A of Compound 1 and geometric isomers of Compound 1 were tested. The results of the tests of the effects of GCS inhibitors on live K-562 cells are shown respectively in Figure 33 A (eliglustat), Figure 33 B (venglustat), Figure 33 C (polymorph A of Compound 1) and Figure 33 D (geometric isomers of Compound 1), and are also summarized in Table 25 below. The results showed that the test compounds (i.e., polymorph A of Compound 1 and geometric isomers of Compound 1) inhibited GCS activity and were more effective than the control compounds eliglustat and venglustat.

[0448] Table 25 Summary of the effects of GCS inhibitors on live K-562 cells

[0449] Compound <![CDATA[IC 50 (nM)]]> Elubrixostat 63.11 Venglustat 511.2 Polymorph A of Compound 1 6.15 Geometric isomer of Compound 1 14.77

[0450] Test of the effect of GCS inhibitors using L6 cell lysates

[0451] To confirm the inhibitory activity of the test compounds on GCS from different species, cells from rats were used. First, the presence of GCS activity was verified using different amounts of L6 cell lysates ( Figure 34 , Table 26). Next, the inhibitory effect of polymorph A of Compound 1 on GCS activity from L6 cell lysates was tested. Polymorph A of Compound 1 showed effective inhibitory effect on GCS derived from rat skeletal muscle (Tables 27 and 28). When 30 μg of cell lysate was used, the inhibitory level of polymorph A of Compound 1 on GCS was > 98%, which was consistent with the fact that polymorph A of Compound 1 had effective inhibitory activity on rat GCS enzyme.

[0452] Table 26 Summary of GCS activity of L6 cell lysates

[0453]

[0454] Summary of the effects of GCS inhibitors on L6 cell lysates - control compounds

[0455]

[0456] Table 28 Summary of the effects of GCS inhibitors on L6 cell lysates - test compounds

[0457]

[0458] These results indicate that, compared with the control compounds (elinurad and venglustat), the test compounds (polymorph A of compound 1 and the geometric isomer of compound 1) have stronger inhibitory activity against GCS. These test compounds also showed similar inhibitory activity against GCS enzymes derived from different organisms (including rats, dogs, and humans).

Claims

1. Polymorph A of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, wherein the polymorph A is characterized in that its X-ray powder diffraction (XRPD) pattern comprises one or more peaks selected from the group consisting of: at about 18.883°, about 10.889° and about 21.309° 2θ.

2. The polymorph A according to claim 1, characterized in that its XRPD pattern comprises all peaks selected from the group consisting of: at about 18.883°, about 10.889° and about 21.309° 2θ.

3. The polymorph A according to claim 1, characterized in that its XRPD pattern further comprises one or more peaks selected from the group consisting of: at about 14.974°, about 24.160°, about 18.663°, about 17.109°, about 7.518°, about 20.950°, about 19.602°, about 27.784° and about 28.281° 2θ.

4. The polymorph A according to claim 1, characterized in that its XRPD pattern further comprises two or more peaks selected from the group consisting of: at about 14.974°, about 24.160°, about 18.663°, about 17.109°, about 7.518°, about 20.950°, about 19.602°, about 27.784° and about 28.281° 2θ.

5. The polymorph A according to claim 1, characterized in that its XRPD pattern further comprises three or more peaks selected from the group consisting of: at about 14.974°, about 24.160°, about 18.663°, about 17.109°, about 7.518°, about 20.950°, about 19.602°, about 27.784° and about 28.281° 2θ.

6. The polymorph A according to claim 1, characterized in that its XRPD pattern further comprises all peaks selected from the group consisting of: at about 14.974°, about 24.160°, about 18.663°, about 17.109°, about 7.518°, about 20.950°, about 19.602°, about 27.784° and about 28.281° 2θ.

7. The polymorph A according to claim 3, characterized in that its XRPD pattern further comprises one or more peaks selected from the group consisting of: at about 17.494°, about 14.743°, about 22.386°, about 31.979°, about 21.759°, about 25.017°, about 30.623°, about 10.301°, about 26.760°, about 21.536°, about 19.337°, about 30.103°, about 33.310°, about 31.467°, about 23.309°, about 25.781° and about 27.362° 2θ.

8. The polymorph A according to claim 3, characterized in that Its XRPD pattern further includes two or more peaks selected from the group consisting of: at about 17.494°, about 14.743°, about 22.386°, about 31.979°, about 21.759°, about 25.017°, about 30.623°, about 10.301°, about 26.760°, about 21.536°, about 19.337°, about 30.103°, about 33.310°, about 31.467°, about 23.309°, about 25.781° and about 27.362° 2θ.

9. The crystalline form A according to claim 3, wherein its XRPD pattern further includes three or more peaks selected from the group consisting of: at about 17.494°, about 14.743°, about 22.386°, about 31.979°, about 21.759°, about 25.017°, about 30.623°, about 10.301°, about 26.760°, about 21.536°, about 19.337°, about 30.103°, about 33.310°, about 31.467°, about 23.309°, about 25.781° and about 27.362° 2θ.

10. The crystalline form A according to claim 3, wherein its XRPD pattern further includes all peaks selected from the group consisting of: at about 17.494°, about 14.743°, about 22.386°, about 31.979°, about 21.759°, about 25.017°, about 30.623°, about 10.301°, about 26.760°, about 21.536°, about 19.337°, about 30.103°, about 33.310°, about 31.467°, about 23.309°, about 25.781° and about 27.362° 2θ.

11. The crystalline form A according to any one of the preceding claims, wherein its XRPD pattern includes all peaks selected from the group consisting of: 。 12. The crystalline form A according to any one of the preceding claims, wherein the XRPD pattern is substantially as shown in Figure 1.

13. The crystalline form A according to any one of the preceding claims, wherein its differential scanning calorimetry (DSC) curve includes an endotherm at about 206.9 °C.

14. The crystalline form A according to claim 13, wherein the DSC curve is substantially as shown in Figure 3.

15. A crystalline form B of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, wherein the crystalline form B is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of: at about 6.244°, about 18.679° and about 12.445° 2θ.

16. The crystalline form B according to claim 15, wherein its XRPD pattern includes all peaks selected from the group consisting of: at about 6.244°, about 18.679° and about 12.445° 2θ.

17. The crystalline form B according to claim 15, wherein its XRPD pattern further comprises one or more peaks selected from the group consisting of: at about 17.239°, about 12.832°, about 23.559°, about 15.252°, about 22.410°, about 9.344° and about 25.722° 2θ.

18. The crystalline form B according to claim 15, wherein its XRPD pattern further comprises two or more peaks selected from the group consisting of: at about 17.239°, about 12.832°, about 23.559°, about 15.252°, about 22.410°, about 9.344° and about 25.722° 2θ.

19. The crystalline form B according to claim 15, wherein its XRPD pattern further comprises three or more peaks selected from the group consisting of: at about 17.239°, about 12.832°, about 23.559°, about 15.252°, about 22.410°, about 9.344° and about 25.722° 2θ.

20. The crystalline form B according to claim 15, wherein its XRPD pattern further comprises all the peaks selected from the group consisting of: at about 17.239°, about 12.832°, about 23.559°, about 15.252°, about 22.410°, about 9.344° and about 25.722° 2θ.

21. The crystalline form B according to claim 17, wherein its XRPD pattern further comprises one or more peaks selected from the group consisting of: at about 20.354°, about 16.366°, about 24.731°, about 20.581°, about 28.664°, about 27.044°, about 35.758°, about 17.905°, about 16.087°, about 36.194°, about 34.867°, about 24.984°, about 10.802°, about 29.902° and about 32.584° 2θ.

22. The crystalline form B according to claim 17, wherein its XRPD pattern further comprises two or more peaks selected from the group consisting of: at about 20.354°, about 16.366°, about 24.731°, about 20.581°, about 28.664°, about 27.044°, about 35.758°, about 17.905°, about 16.087°, about 36.194°, about 34.867°, about 24.984°, about 10.802°, about 29.902° and about 32.584° 2θ.

23. The crystalline form B according to claim 17, wherein Its XRPD pattern further includes three or more peaks selected from the group consisting of: at about 20.354°, about 16.366°, about 24.731°, about 20.581°, about 28.664°, about 27.044°, about 35.758°, about 17.905°, about 16.087°, about 36.194°, about 34.867°, about 24.984°, about 10.802°, about 29.902° and about 32.584° 2θ.

24. The crystalline form B according to claim 17, wherein its XRPD pattern further includes all peaks selected from the group consisting of: at about 20.354°, about 16.366°, about 24.731°, about 20.581°, about 28.664°, about 27.044°, about 35.758°, about 17.905°, about 16.087°, about 36.194°, about 34.867°, about 24.984°, about 10.802°, about 29.902° and about 32.584° 2θ.

25. The crystalline form B according to any one of claims 15 to 24, wherein its XRPD pattern includes all peaks selected from the group consisting of: 。 26. The crystalline form B according to any one of claims 15 to 25, wherein the XRPD pattern is substantially as shown in Figure 5.

27. The crystalline form B according to any one of claims 15 to 26, wherein its DSC curve includes endotherms at about 178.3 °C, about 179.7 °C, about 196.6 °C, about 198.7 °C and / or about 203.7 °C.

28. The crystalline form B according to claim 27, wherein the DSC curve is substantially as shown in Figure 7.

29. A crystalline form C of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, wherein the crystalline form C is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of: at about 6.656°, about 19.994° and about 26.732° 2θ.

30. The crystalline form C according to claim 29, wherein its XRPD pattern includes all peaks selected from the group consisting of: at about 6.656°, about 19.994° and about 26.732° 2θ.

31. The crystalline form C according to claim 29, wherein its XRPD pattern further includes one or more peaks selected from the group consisting of: at about 18.350°, about 20.753°, about 22.176°, about 15.941°, about 20.450°, about 13.320° and about 17.687° 2θ.

32. The crystalline form C according to claim 29, wherein Its XRPD pattern further includes two or more peaks selected from the group consisting of about 18.350°, about 20.753°, about 22.176°, about 15.941°, about 20.450°, about 13.320° and about 17.687° 2θ.

33. Polymorph C according to claim 29, wherein its XRPD pattern further includes three or more peaks selected from the group consisting of about 18.350°, about 20.753°, about 22.176°, about 15.941°, about 20.450°, about 13.320° and about 17.687° 2θ.

34. Polymorph C according to claim 29, wherein its XRPD pattern further includes all peaks selected from the group consisting of about 18.350°, about 20.753°, about 22.176°, about 15.941°, about 20.450°, about 13.320° and about 17.687° 2θ.

35. Polymorph C according to any one of claims 29 to 34, wherein its XRPD pattern includes all peaks selected from the group consisting of 。 36. Polymorph C according to any one of claims 29 to 35, wherein the XRPD pattern is substantially as shown in Figure 9.

37. Polymorph C according to any one of claims 29 to 36, wherein the DSC curve is substantially as shown in Figure 11.

38. A polymorph D of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, wherein the polymorph D is characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 16.956°, about 5.883° and about 22.181° 2θ.

39. Polymorph D according to claim 38, wherein its XRPD pattern includes all peaks selected from the group consisting of about 16.956°, about 5.883° and about 22.181° 2θ.

40. Polymorph D according to claim 38, wherein its XRPD pattern further includes one or more peaks selected from the group consisting of about 17.663°, about 13.291°, about 22.909°, about 20.718°, about 26.900° and about 18.113° 2θ.

41. Polymorph D according to claim 38, wherein its XRPD pattern further includes two or more peaks selected from the group consisting of about 17.663°, about 13.291°, about 22.909°, about 20.718°, about 26.900° and about 18.113° 2θ.

42. Polymorph D according to claim 38, wherein its XRPD pattern further includes three or more peaks selected from the group consisting of about 17.663°, about 13.291°, about 22.909°, about 20.718°, about 26.900° and about 18.113° 2θ.

43. Polymorph D according to claim 38, wherein its XRPD pattern further comprises all peaks selected from the group consisting of: at about 17.663°, about 13.291°, about 22.909°, about 20.718°, about 26.900° and about 18.113° 2θ.

44. Polymorph D according to claim 40, wherein its XRPD pattern further comprises one or more peaks selected from the group consisting of: at about 20.205°, about 11.727°, about 21.756°, about 22.617°, about 20.956°, about 29.635°, about 24.990°, about 12.566°, about 10.080°, about 24.078°, about 9.761°, about 19.581°, about 27.280°, about 25.637° and about 12.754° 2θ.

45. Polymorph D according to claim 40, wherein its XRPD pattern further comprises two or more peaks selected from the group consisting of: at about 20.205°, about 11.727°, about 21.756°, about 22.617°, about 20.956°, about 29.635°, about 24.990°, about 12.566°, about 10.080°, about 24.078°, about 9.761°, about 19.581°, about 27.280°, about 25.637° and about 12.754° 2θ.

46. Polymorph D according to claim 40, wherein its XRPD pattern further comprises three or more peaks selected from the group consisting of: at about 20.205°, about 11.727°, about 21.756°, about 22.617°, about 20.956°, about 29.635°, about 24.990°, about 12.566°, about 10.080°, about 24.078°, about 9.761°, about 19.581°, about 27.280°, about 25.637° and about 12.754° 2θ.

47. Polymorph D according to claim 40, wherein its XRPD pattern further comprises all peaks selected from the group consisting of: at about 20.205°, about 11.727°, about 21.756°, about 22.617°, about 20.956°, about 29.635°, about 24.990°, about 12.566°, about 10.080°, about 24.078°, about 9.761°, about 19.581°, about 27.280°, about 25.637° and about 12.754° 2θ.

48. Polymorph D according to any one of claims 38 to 47, wherein its XRPD pattern comprises all peaks selected from the group consisting of:

49. Polymorph D according to any one of claims 38 to 48, wherein the XRPD pattern is substantially as shown in Figure 13.

50. Polymorph D according to any one of claims 38 to 49, wherein its DSC curve comprises an endotherm at about 207.4 °C.

51. The crystalline form D according to claim 50, wherein the DSC curve is substantially as shown in Figure 15.

52. A crystalline form E of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, wherein the crystalline form E is characterized in that its XRPD pattern comprises one or more peaks selected from the group consisting of: at about 18.089°, about 26.315° and about 19.775° 2θ.

53. The crystalline form E according to claim 52, characterized in that its XRPD pattern comprises all the peaks selected from the group consisting of: at about 18.089°, about 26.315° and about 19.775° 2θ.

54. The crystalline form E according to claim 52, characterized in that its XRPD pattern further comprises one or more peaks selected from the group consisting of: at about 24.177°, about 20.803°, about 16.018°, about 21.429° and about 14.608° 2θ.

55. The crystalline form E according to claim 52, characterized in that its XRPD pattern further comprises two or more peaks selected from the group consisting of: at about 24.177°, about 20.803°, about 16.018°, about 21.429° and about 14.608° 2θ.

56. The crystalline form E according to claim 52, characterized in that its XRPD pattern further comprises three or more peaks selected from the group consisting of: at about 24.177°, about 20.803°, about 16.018°, about 21.429° and about 14.608° 2θ.

57. The crystalline form E according to claim 52, characterized in that its XRPD pattern further comprises all the peaks selected from the group consisting of: at about 24.177°, about 20.803°, about 16.018°, about 21.429° and about 14.608° 2θ.

58. The crystalline form E according to claim 54, characterized in that its XRPD pattern further comprises one or more peaks selected from the group consisting of: at about 21.314°, about 7.794°, about 20.007°, about 8.744° and about 15.573° 2θ.

59. The crystalline form E according to claim 54, characterized in that its XRPD pattern further comprises two or more peaks selected from the group consisting of: at about 21.314°, about 7.794°, about 20.007°, about 8.744° and about 15.573° 2θ.

60. The crystalline form E according to claim 54, characterized in that its XRPD pattern further comprises three or more peaks selected from the group consisting of: at about 21.314°, about 7.794°, about 20.007°, about 8.744° and about 15.573° 2θ.

61. The crystalline form E according to claim 54, characterized in that Its XRPD pattern further includes all peaks selected from the group consisting of about 21.314°, about 7.794°, about 20.007°, about 8.744° and about 15.573° 2θ.

62. Polymorph E according to any one of claims 52 to 61, characterized in that its XRPD pattern includes all peaks selected from the group consisting of:

63. Polymorph E according to any one of claims 52 to 62, wherein the XRPD pattern is substantially as shown in Figure 17.

64. The polymorph according to any one of the preceding claims, wherein the polymorph is substantially pure.

65. Polymorph A of the L-malate salt of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of about 19.349°, about 22.304° and about 20.078° 2θ.

66. Polymorph A of the L-malate salt according to claim 65, characterized in that its XRPD pattern includes all peaks selected from the group consisting of about 19.349°, about 22.304° and about 20.078° 2θ.

67. Polymorph A of the L-malate salt according to claim 65, characterized in that its XRPD pattern further includes one or more peaks selected from the group consisting of about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597° and about 17.581° 2θ.

68. Polymorph A of the L-malate salt according to claim 65, characterized in that its XRPD pattern further includes two or more peaks selected from the group consisting of about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597° and about 17.581° 2θ.

69. Polymorph A of the L-malate salt according to claim 65, characterized in that its XRPD pattern further includes three or more peaks selected from the group consisting of about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597° and about 17.581° 2θ.

70. Polymorph A of the L-malate salt according to claim 65, characterized in that Its XRPD pattern further includes all peaks selected from the group consisting of: at about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597° and about 17.581° 2θ.

71. The crystalline form A of L-malate according to claim 67, wherein its XRPD pattern further includes one or more peaks selected from the group consisting of: at about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704° and about 21.715° 2θ.

72. The crystalline form A of L-malate according to claim 67, wherein its XRPD pattern further includes two or more peaks selected from the group consisting of: at about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704° and about 21.715° 2θ.

73. The crystalline form A of L-malate according to claim 67, wherein its XRPD pattern further includes three or more peaks selected from the group consisting of: at about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704° and about 21.715° 2θ.

74. The crystalline form A of L-malate according to claim 67, wherein its XRPD pattern further includes all peaks selected from the group consisting of: at about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704° and about 21.715° 2θ.

75. The crystalline form A of L-malate according to any one of claims 65 to 74, wherein Its XRPD pattern includes all peaks selected from the group consisting of: at about 19.349°, about 22.304°, about 20.078°, about 24.489°, about 22.982°, about 18.186°, about 19.083°, about 6.369°, about 12.702°, about 24.937°, about 28.749°, about 24.407°, about 23.772°, about 23.597°, about 17.581°, about 16.263°, about 31.742°, about 18.334°, about 25.587°, about 3.208°, about 8.765°, about 9.529°, about 20.434°, about 30.739°, about 18.878°, about 26.992°, about 21.941°, about 20.948°, about 36.810°, about 34.693°, about 32.704°, about 21.715°, about 36.432°, about 31.492°, about 27.976°, about 37.328°, about 17.975°, about 38.681°, about 15.886°, about 13.678°, about 32.223°, about 26.405°, about 27.514°, about 31.082°, about 14.561°, about 39.38°, about 35.442°, about 26.035°, about 26.739°, about 35.592°, about 11.409°, about 10.2°, about 8.962°, about 34.334°, about 28.307°, about 35.984°, about 29.228°, about 39.705°, about 33.715° and about 30.135° 2θ.

76. The crystalline form A of L-malate according to any one of claims 65 to 75, wherein the XRPD pattern is substantially as shown in Figure 18.

77. The crystalline form A of L-malate according to any one of claims 65 to 76, characterized in that its DSC curve includes an endotherm at about 160.3 °C.

78. The crystalline form A of L-malate according to any one of claims 77, wherein the DSC curve is substantially as shown in Figure 20.

79. A crystalline form A of L-tartrate of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide, characterized in that its XRPD pattern includes one or more peaks selected from the group consisting of: at about 18.564°, about 21.685° and about 18.774° 2θ.

80. The crystalline form A of L-tartrate according to claim 79, characterized in that its XRPD pattern includes all peaks selected from the group consisting of: at about 18.564°, about 21.685° and about 18.774° 2θ.

81. The crystalline form A of L-tartrate according to claim 79, characterized in that Its XRPD pattern further includes one or more peaks selected from the group consisting of about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965° and about 24.193° 2θ.

82. The crystalline form A of L-tartrate according to claim 79, wherein its XRPD pattern further includes two or more peaks selected from the group consisting of about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965° and about 24.193° 2θ.

83. The crystalline form A of L-tartrate according to claim 79, wherein its XRPD pattern further includes three or more peaks selected from the group consisting of about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965° and about 24.193° 2θ.

84. The crystalline form A of L-tartrate according to claim 79, wherein its XRPD pattern further includes all of the peaks selected from the group consisting of about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965° and about 24.193° 2θ.

85. The crystalline form A of L-tartrate according to claim 81, wherein its XRPD pattern further includes one or more peaks selected from the group consisting of about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641° and about 19.08° 2θ.

86. The crystalline form A of L-tartrate according to claim 81, wherein its XRPD pattern further includes two or more peaks selected from the group consisting of about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641° and about 19.08° 2θ.

87. The crystalline form A of L-tartrate according to claim 81, wherein Its XRPD pattern further includes three or more peaks selected from the group consisting of: about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641° and about 19.08° 2θ.

88. The crystalline form A of L-tartrate according to claim 81, wherein its XRPD pattern further includes all peaks selected from the group consisting of: about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641° and about 19.08° 2θ.

89. The crystalline form A of L-tartrate according to any one of claims 79 to 88, wherein its XRPD pattern includes all peaks selected from the group consisting of: about 18.564°, about 21.685°, about 18.774°, about 24.834°, about 22.437°, about 37.586°, about 24.428°, about 20.965°, about 24.193°, about 17.789°, about 3.135°, about 19.49°, about 25.427°, about 31.744°, about 16.413°, about 9.279°, about 26.623°, about 12.36°, about 27.826°, about 14.438°, about 12.221°, about 24.045°, about 31.932°, about 19.234°, about 28.295°, about 20.641°, about 19.08°, about 10.165°, about 33.761°, about 29.071°, about 25.916°, about 16.943°, about 32.967°, about 23.691°, about 22.788°, about 34.232°, about 35.17°, about 34.812°, about 27.271°, about 32.24°, about 39.082°, about 30.774°, about 15.453°, about 30.534°, about 31.183°, about 37.181°, about 35.805°, about 28.554°, about 29.645°, about 39.789°, about 38.258°, about 10.432°, about 13.968° and about 36.619° 2θ.

90. The crystalline form A of L-tartrate according to any one of claims 79 to 89, wherein the XRPD pattern is substantially as shown in Figure 22.

91. The crystalline form A of L-tartrate according to any one of claims 79 to 90, characterized in that its DSC curve includes an endotherm at about 189.9 °C.

92. The crystalline form A of L-tartrate according to claim 91, wherein the DSC curve is substantially as shown in Figure 24.

93. A pharmaceutical composition comprising (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, wherein (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide comprises a crystalline form selected from the group consisting of: crystalline form A according to any one of claims 1 to 14, crystalline form B according to any one of claims 15 to 28, crystalline form C according to any one of claims 29 to 37, crystalline form D according to any one of claims 38 to 51, crystalline form E according to any one of claims 52 to 63, the crystalline form according to claim 64, crystalline form A of L-malate according to any one of claims 65 to 78, and crystalline form A of L-tartrate according to any one of claims 79 to 92.

94. The pharmaceutical composition according to claim 93, wherein at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5%, at least about 99.9%, at least about 99.99% of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide has the crystalline form A.

95. The pharmaceutical composition according to claim 93, wherein at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5%, at least about 99.9%, at least about 99.99% of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide has the crystalline form B.

96. The pharmaceutical composition according to claim 93, wherein at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5%, at least about 99.9%, at least about 99.99% of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide has the crystalline form C.

97. The pharmaceutical composition according to claim 93, wherein at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5%, at least about 99.9%, at least about 99.99% of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide has the crystalline form D.

98. The pharmaceutical composition according to claim 93, wherein at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5%, at least about 99.9%, at least about 99.99% of (E)-N-((1R,2R)-1-(3-chloro-4-cyclopropoxyphenyl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)-2-(6-chloronaphthalen-2-yl)-2-(hydroxyimino)acetamide has the crystalline form E.

99. The pharmaceutical composition according to claim 93, which further comprises an additional therapeutic agent.

100. A method for preparing a pharmaceutical composition, the method comprising mixing a crystalline form according to any one of claims 1 to 64, or crystalline form A of L-malate according to any one of claims 65 to 78, or crystalline form A of L-tartrate according to any one of claims 79 to 92 with a pharmaceutically acceptable excipient.

101. Use of a crystalline form according to any one of claims 1 to 64, or crystalline form A of L-malate according to any one of claims 65 to 78, or crystalline form A of L-tartrate according to any one of claims 69 to 92, or a pharmaceutical composition according to any one of claims 93 to 99 in the preparation of a medicament for: (a) preventing or treating a disease, disorder or condition associated with abnormal activity / level of the enzyme GCS; (b) preventing or treating a GCS-mediated disease, disorder or condition; (c) inhibiting the activity / level of the enzyme GCS; or (d) non-therapeutically inhibiting the activity / level of the GCS enzyme in vitro.

102. The use according to claim 101, wherein the disease, disorder or condition associated with the abnormal activity / level of the enzyme GCS, or the GCS-mediated disease, disorder or condition is selected from the group consisting of: glycosphingolipid storage diseases (e.g., Tay-Sachs disease, Sandhoff disease, GM1 gangliosidosis and Fabry disease); diseases associated with glycosphingolipid accumulation (e.g., Gaucher disease); diseases causing renal hypertrophy or hyperplasia, such as diabetic nephropathy; diseases causing hyperglycemia or hyperinsulinemia; cancers with abnormal glycosphingolipid synthesis; infectious diseases caused by organisms that use cell surface glycosphingolipids as receptors or for which the synthesis of glucosylceramide is essential or important; metabolic disorders, such as atherosclerosis, polycystic kidney disease, renal hypertrophy and diabetes; obesity; cancers, such as breast cancer, renal adenocarcinoma, brain cancer, neuroblastoma, lung cancer, intestinal cancer, pancreatic cancer and prostate cancer; neuronal disorders; neuronal injury; inflammatory diseases or disorders (e.g., rheumatoid arthritis, Crohn's disease, asthma and sepsis); pain (e.g., neuropathic pain, inflammatory pain, headache, somatic pain, visceral pain, referred pain); cognitive disorders (e.g., agnosia; amnesia; aphasia; apraxia; delirium; dementia, including AIDS dementia complex, Binswanger's disease, Lewy body dementia, frontotemporal dementia, mild cognitive impairment, multi-infarct dementia, Pick's disease, semantic dementia, Alzheimer's disease and vascular dementia; and learning disorders, including Asperger syndrome, attention deficit disorder, attention deficit hyperactivity disorder, autism, childhood disintegrative disorder and Rett syndrome); neurodegenerative disorders (such as Alzheimer's disease, corticobasal degeneration, Creutzfeldt-Jakob disease, frontotemporal lobar degeneration, Huntington's disease, multiple sclerosis, normal pressure hydrocephalus, organic chronic brain syndrome, Parkinson's disease, Pick's disease, progressive supranuclear palsy and Alzheimer's (Alzheimer type), glomerular diseases) and non-alcoholic fatty liver disease (NALD).

103. The use according to claim 102, wherein the disease, disorder or condition is Tay-Sachs disease, Sandhoff disease, GM1 gangliosidosis, Fabry disease, Gaucher disease or polycystic kidney disease.

104. A method of inhibiting GCS activity, the method comprising administering to a subject in need thereof an effective amount of a crystalline form according to any one of claims 1 to 64, or crystalline form A of L-malate according to any one of claims 65 to 78, or crystalline form A of L-tartrate according to any one of claims 79 to 92, or a pharmaceutical composition according to any one of claims 93 to 99.

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