(2S, 5R)-5-(2-chlorphenyl)-1-(2apos; -methoxy-[1, 1apos; polymorphic substance of-biphenyl]-4-carbonyl) pyrrolidine-2-carboxylic acid and preparation method thereof

Abstract

The invention relates to a crystal form I and a crystal form II of (2S, 5R)-5-(2-chlorphenyl)-1-(2 '-methoxy-[1, 1'-biphenyl]-4-carbonyl) pyrrolidine-2-carboxylic acid and a preparation method thereof.

Description

Field of the Invention

[0001] The present invention relates to crystal forms of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid, namely, Crystal Form I and Crystal Form II. The present invention also provides methods for preparing Crystal Form I and Crystal Form II. Crystal Form I and Crystal Form II can be used to treat diseases associated with or mediated by free fatty acid receptor 2 (FFAR2) (also known as the GPR43 receptor). Background of the Invention

[0003] As reported in WO2015 / 078949, the pyrrolidine carboxylic acid derivatives disclosed in WO2011 / 073376 are FFAR2 agonists and can be used to treat inflammatory diseases. In particular, there is evidence that (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (hereinafter referred to as Compound (1)) is of particular interest for the treatment of inflammatory diseases. WO2021 / 250174 reports a method for the scalable manufacture of Compound (1) and its corresponding sodium salt (Compound (1.Na)).

[0004]

[0005] In all of the above references, Compound (1) and its sodium salt (1.Na) were obtained in amorphous form. However, the main disadvantage of the amorphous forms of Compound (1) and (1.Na) is their strong hygroscopicity, which causes many problems in the manufacture, handling, storage, and use of the compound in solid dosage forms.

[0006] In addition, for the active ingredient (i.e., active pharmaceutical ingredient (API)) to be used in pharmaceutical formulations, the amount of residual solvent in the API needs to be as low as possible to meet the pharmacopoeial requirements for residual solvents. In fact, the presence of trace organic solvents can be harmful, and therefore, the solvates of the API are not recommended for use in pharmaceuticals. In addition, even the hydrates of the API are preferably avoided because they tend to be hygroscopic and may require more control during the manufacture, storage, and use of the formulation. Hydrates generally have lower water solubility than the anhydrous forms, making it more difficult to achieve an absorbable dose.

[0007] Therefore, it is necessary to provide a non-solvated and non-hygroscopic form of Compound (1).

[0008] Studying polymorphism during the drug development process is common practice. However, the formation of polymorphs, their composition (i.e., molecules existing alone or in the form of salts and / or solvates), and their properties are unpredictable.

[0009] Different crystal forms of the same compound can have different physical properties, such as different packing, thermodynamic, spectroscopic, kinetic, surface, and mechanical properties. This can lead to differences in ease of processing, stability, the possibility of formulation into dosage forms, dissolution rate, and thus bioavailability. Although general methods for API crystal form screening are known, it has been well established that it is not possible to predict whether a given compound will exhibit polymorphism. In addition, even when crystal forms exist, predictions about the properties of the crystal forms and how they differ from each other remain elusive.

[0010] During the course of the applicant's research, several polymorphs of compound (1), namely at least 10 forms, were identified. Unfortunately, most of them are solvated forms, that is, the polymorphs of compound (1) contain one or more solvent molecules. Among all the identified polymorphs, only Form I and Form II were shown to be non-solvated and non-hygroscopic. In addition, Form I was shown to be particularly stable, especially compared to Form II.

[0011] Accordingly, the present invention provides Form I and Form II of compound (1), which are anhydrous and non-solvated polymorphs and are non-hygroscopic. Advantageously, Form I is particularly stable compared to Form II.

[0012] Overview

[0013] Accordingly, the present invention relates to crystal forms of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid: Form I or Form II, wherein:

[0014] Form I is characterized in that its XRPD pattern includes peaks at 2θ angular values of 13.5°, 14.0°, 14.8°, 16.0°, and 18.0°; and

[0015] Form II is characterized in that its XRPD pattern includes peaks at 2θ angular values of 10.8°, 12.1°, 12.4°, and 22.3°.

[0016] In one embodiment, the XRPD pattern of Form I includes peaks at 2θ angular values of 7.2°, 12.8°, 13.5°, 14.0°, 14.5°, 14.8°, 16.0°, 16.7°, 17.4°, 18.0°, 18.9°, 19.9°, 20.4°, and 23.2°. In one embodiment, Form I has an XRPD pattern substantially as Figure 1 shown.

[0017] In one embodiment, the DSC thermogram of Form I shows an endothermic peak with a peak temperature of about 183 °C and a start temperature of about 180 °C.

[0018] In one embodiment, the unit cell parameters of Form I are equal to:

[0019] Unit cell dimensions:

[0020]

[0021] α, β, γ = 90°

[0022] Space group: P2 1 2 1 2 1

[0023] Number of molecules per unit cell: 4

[0024] Unit cell volume:

[0025] Density (calculated): 1.19 g / cm 3 .

[0026] In one embodiment, the XRPD pattern of Form II includes peaks at 2θ values of 10.8°, 12.1°, 12.4°, 15.3°, 16.2°, 18.5°, 19.5°, 20.7°, 21.5° and 22.3°. In one embodiment, Form II has an XRPD pattern substantially as Figure 5 shown.

[0027] In one embodiment, the DSC thermogram of Form II shows a first endothermic peak with a peak temperature of about 110 °C starting at about 100 °C; an exothermic peak with a peak temperature of about 149 °C and starting at about 134 °C; and a second endothermic peak with a peak temperature of about 181 °C starting at about 179 °C.

[0028] The present invention also provides a method for preparing Form I, which comprises:

[0029] 1) Dissolving (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid in an ethanol / water mixture at a temperature of at most 80 °C, wherein the ethanol / water ratio is from 100 / 0 to 5 / 95;

[0030] 2) If necessary, adding water to bring the ethanol / water ratio to from 95 / 5 to 5 / 95, wherein the water is preferably heated at the same temperature as the ethanol used in step 1); and

[0031] 3) Cooling the mixture, preferably to a temperature of 0 °C to 10 °C, and maintaining this temperature for a period suitable for recovering Form I.

[0032] The present invention also provides a method for preparing polymorph II, which comprises:

[0033] 1) Dissolving (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid in acetonitrile at a temperature of at most 80 °C; and

[0034] 2) Cooling the mixture of step 1), preferably to room temperature, and maintaining this temperature for a period suitable for recovering polymorph II.

[0035] The present invention also provides a pharmaceutical composition comprising the polymorph according to the present invention and at least one pharmaceutically acceptable carrier.

[0036] The present invention also relates to the polymorph according to the present invention for use as a medicament.

[0037] The present invention also relates to a crystalline form according to the present invention, which is used for treating inflammatory diseases, wherein the inflammatory diseases are selected from rheumatoid arthritis; inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut dysbiosis, including antibiotic-induced dysbiosis, which leads to bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia, etc.); Paget's disease; osteoporosis; multiple myeloma; uveitis; acute myeloid leukemia, chronic myeloid leukemia; pancreatic β-cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritis conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; anaphylaxis; contact dermatitis; pancreatitis; allergy; hepatitis, including hepatitis B virus infection; asthma; muscle degeneration; cachexia, such as cachexia secondary to infection or malignancy, cachexia secondary to acquired immunodeficiency syndrome; Reiter's syndrome; type I diabetes; bone resorption disease; graft-versus-host reaction; ischemia-reperfusion injury; brain trauma; multiple sclerosis; autoimmune brain diseases, such as encephalitis and encephalomyelitis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxic shock; gram-negative sepsis; fever and myalgia caused by infection (such as influenza); heartburn;Inflammatory conditions triggered by the release of anorectic gut hormones (e.g., PYY, GLP-1), such as those observed in hyperphagia-related disorders, obesity, type 2 diabetes, etc.; microbiome-related neurological and mood disorders, including autism spectrum disorder, schizophrenia, depression, major depressive disorder, and neurodegenerative diseases characterized by neuroinflammation, including Alzheimer's disease and Parkinson's disease.

[0038] In one embodiment, the inflammatory disease is selected from rheumatoid arthritis; inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis; immune-mediated colitis (IMC); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis; Paget's disease; osteoporosis; multiple myeloma; uveitis; acute myeloid leukemia, chronic myeloid leukemia; pancreatic beta cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritic conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis; asthma; muscle degeneration; cachexia; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; autoimmune brain diseases, encephalitis, encephalomyelitis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; gram-negative sepsis; fever and myalgia caused by infection; heartburn; inflammatory conditions triggered by the release of anorectic gut hormones; microbiome-related neurological and mood disorders, and neurodegenerative diseases characterized by neuroinflammation.

[0039] In one embodiment, the inflammatory disease is selected from inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, colitis, collagenous colitis, lymphocytic colitis, immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), pouchitis, celiac disease, irritable bowel syndrome, gut microbiota dysbiosis, type 1 diabetes, multiple sclerosis, autoimmune brain diseases such as encephalitis and encephalomyelitis. In one embodiment, the inflammatory disease is selected from inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, colitis, collagenous colitis, lymphocytic colitis, immune-related enterocolitis, immune-mediated colitis (IMC), pouchitis, celiac disease, irritable bowel syndrome, gut microbiota dysbiosis, type 1 diabetes, multiple sclerosis, autoimmune brain diseases, encephalitis and encephalomyelitis. In one embodiment, the immune-related enterocolitis is immune-related enterocolitis that occurs as an adverse event of cancer therapy in response to the use of checkpoint inhibitors, preferably the checkpoint inhibitor is an inhibitor of CTLA-4, PD-1, and / or PD-L1. In one embodiment, the immune-mediated colitis (IMC) is immune-mediated colitis that occurs as an adverse event of cancer therapy in response to the use of checkpoint inhibitors, preferably the checkpoint inhibitor is an inhibitor of CTLA-4, PD-1, and / or PD-L1.

[0040] The present invention also relates to the crystalline form according to the present invention, which is used for treating an inflammatory disease, wherein the inflammatory disease is preferably selected from inflammatory bowel disease (IBD), colitis, collagenous colitis, lymphocytic colitis, immune-related enterocolitis, pouchitis, celiac disease, irritable bowel syndrome, and gut microbiota dysbiosis.

[0041] The present invention also relates to the crystalline form according to the present invention, which is used for treating cancer, wherein the cancer is preferably selected from lung cancer, non-small cell lung cancer, small cell lung cancer, breast cancer, prostate cancer, ovarian cancer, endometrial cancer, vaginal cancer, testicular cancer, cervical cancer, bladder cancer, head and neck cancer, kidney cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, thyroid cancer, gastrointestinal cancer, colorectal cancer, gastric cancer, colon cancer, liver cancer, leukemia, lymphoma, skin cancer, melanoma, multiple myeloma, glioma, glioblastoma, mesothelioma, retinoblastoma, sarcoma, Ewing's sarcoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, bone cancer, and cardiac cancer.

[0042] Definition

[0043] In the present invention, the following terms have the following meanings:

[0044] "About" before a number means plus or minus 10% of the value of that number, preferably plus or minus 5% of the value of that number.

[0045] "Administration" or its variants (e.g., "administering") means providing an active agent or ingredient, either alone or as part of a pharmaceutically acceptable composition, to a patient to be treated or prevented from a disorder, symptom or disease.

[0046] "Amorphous" or "non-crystalline", when used to describe a substance, refers to the solid state of the substance and means that the substance is arranged in an indefinite geometric shape, i.e., in the form of an irregular array, in contrast to the crystalline form. The amorphous state of a substance can be determined by, for example, X-ray diffraction.

[0047] "Crystalline", when used to describe a substance, refers to the solid state of the substance and means that the substance is arranged in a definite geometric shape, i.e., in the form of a regular array. The crystalline state of a substance can be determined by, for example, X-ray diffraction.

[0048] "Crystalline form", "polymorphic form" or "polymorph" are used interchangeably herein and have the same meaning unless otherwise specified. When used to describe a substance, "crystalline form", "polymorphic form" or "polymorph" refers to the three-dimensional structure in which the substance is organized in the solid state, in contrast to its amorphous form. A substance can have several crystalline forms. Each crystalline form of a substance exhibits different physical properties.

[0049] The terms "crystalline form", "polymorphic form" or "polymorph" can refer only to the crystalline solid state of a substance or can refer to the crystalline solid state of the substance in its salt and / or solvate forms.

[0050] "Non-solvated crystalline form" refers to the crystalline form of a substance that is substantially free of solvent (e.g., less than 1% by weight of solvent, based on the total weight of the substance), which means that there is substantially no solvent molecule (stoichiometric or non-stoichiometric) inside the crystal assembly. "Anhydrous form" is a special case of "non-solvated form" where the solvent is water.

[0051] "Solvated crystalline form" refers to the crystalline form of a substance that contains solvent molecules in stoichiometric or non-stoichiometric proportions relative to the substance.

[0052] "Patient" means a mammal, more preferably a human, who is waiting to receive or is receiving medical care or who is the subject of a medical procedure, either currently or in the future.

[0053] "Pharmaceutically acceptable" means that the components of a pharmaceutical composition are compatible with each other and not harmful to the subject to which it is administered.

[0054] "Pharmaceutically acceptable carrier" means a substance that does not produce adverse, allergic, or other unwanted reactions when administered to an animal (preferably a human). It includes any and all inactive substances, such as solvents, co-solvents, antioxidants, surfactants, stabilizers, emulsifiers, buffers, pH regulators, preservatives (or conserving agents), antibacterial and antifungal agents, isotonifiers, granulating or binding agents, lubricants, disintegrants, glidants, diluents or fillers, adsorbents, dispersants, suspending agents, coating agents, bulking agents, gelatin (for soft and hard capsules), release agents, absorption retardants, sweetening agents, flavoring agents, etc. For human administration, the formulation should meet the sterility, pyrogenicity, general safety, and purity standards required by regulatory agencies (such as the FDA office or EMA).

[0055] "Therapeutically effective amount" means the amount or dose of an active ingredient that is intended to achieve the following purposes without causing significant negative or adverse side effects to the subject: (1) delaying or preventing the onset of a disease in the subject; (2) reducing the severity or incidence of the disease; (3) slowing or stopping the progression, exacerbation, or worsening of one or more symptoms of a disease affecting the subject; (4) causing a reduction in the symptoms of a disease affecting the subject; or (5) curing a disease affecting the subject. A therapeutically effective amount can be administered before the onset of the disease to have a preventive or prophylactic effect. Optionally or additionally, a therapeutically effective amount can be administered after the start of the disease to have a therapeutic effect.

[0056] As used herein, "treating" or "treatment" means therapeutic treatment, prophylactic (or preventive) treatment, or both therapeutic treatment and prophylactic (or preventive) treatment, where the aim is to prevent, reduce, alleviate, and / or slow down (mitigate) one or more symptoms of a target pathological condition or disease in a subject in need thereof. In one embodiment, "treating" means therapeutic treatment. In another embodiment, "treating" means prophylactic or preventive treatment. In yet another embodiment, "treating" means both prophylactic (or preventive) treatment and therapeutic treatment. In one embodiment, "treating" means therapeutic treatment of one or more symptoms of a disease associated with or mediated by FFAR2.

[0057] "Subject" refers to a mammal, preferably a human. In one embodiment, the subject is diagnosed with a disease. In one embodiment, the subject is a patient, preferably a human patient, who is awaiting or undergoing medical care or has been / is / will be the subject of a medical procedure or is being monitored for the development or progression of a disease. In one embodiment, the subject is a human patient who is receiving treatment and / or being monitored for the development or progression of a disease. In one embodiment, the subject is male. In another embodiment, the subject is female. In one embodiment, the subject is an adult. In another embodiment, the subject is a child.

[0058] Detailed Description

[0059] Accordingly, the present invention relates to two polymorphs of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid, which are non-solvated and non-hygroscopic. These two new crystal forms are respectively referred to as "Form I" and "Form II".

[0060] Characterizations of Forms I and II of the present invention are provided below. Depending on the measurement conditions, equipment, and other routine variables known to those skilled in the art, Forms I and II may exhibit similar but not identical analytical characteristics within a reasonable error range compared to those reported herein.

[0061] Form I

[0062] Accordingly, in one embodiment, the present invention relates to Form I of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid.

[0063] Form I is characterized in that its X-ray powder diffraction (XRPD) pattern includes peaks at 2θ values of 13.5°, 14.0°, 14.8°, 16.0°, and 18.0°.

[0064] The 2θ values provided herein are for measurements conducted under the conditions detailed in Example 2 at room temperature. In one embodiment, an X-ray wavelength of is used to measure the peaks of the XRPD pattern. In one embodiment, the error range of the provided angular values is ±0.2°, preferably ±0.1°.

[0065] In one embodiment, the XRPD pattern of Form I includes peaks at 2θ values of 7.2°, 12.8°, 13.5°, 14.0°, 14.5°, 14.8°, 16.0°, 16.7°, 17.4°, 18.0°, 18.9°, 19.9°, 20.4°, and 23.2°.

[0066] In one embodiment, the XRPD pattern of Form I includes peaks with 2θ values similar to those shown in Table 1.

[0067] Table 1. XRPD pattern of Form I and relative intensities of the reflection peaks (2θ).

[0068]

[0069]

[0070] In one embodiment, Form I has an XRPD pattern substantially as Figure 1 shown.

[0071] In one embodiment, Form I crystallizes in an orthorhombic unit cell structure with unit cell parameters equal to:

[0072] Unit cell dimensions:

[0073]

[0074] α, β, γ = 90°

[0075] Space group: P2 1 2 1 2 1

[0076] Number of molecules per unit cell: 4

[0077] Unit cell volume:

[0078] Density (calculated): 1.19 g / cm 3

[0079] In one embodiment, the differential scanning calorimetry (DSC) thermogram of Form I shows an endothermic peak with a peak temperature of about 183 °C and a start temperature of about 180 °C.

[0080] In one embodiment, Form I has a DSC thermogram substantially as Figure 2 shown.

[0081] In one embodiment, Form I has a thermogravimetric analysis (TGA) thermogram substantially as Figure 3 shown, which does not show significant weight loss at temperatures below 220 °C. Above this temperature, the observed weight loss may be due to evaporation and / or degradation.

[0082] It has been demonstrated that Form I is advantageously non-hygroscopic. In particular, a water adsorption / desorption curve was established for Form I, as Figure 4as reported in the dynamic vapor sorption (DVS) isotherm plots and as detailed in Example 6.

[0083] It has also been demonstrated that Polymorph I is thermodynamically stable. In particular, Polymorph I is thermodynamically stable for at least 6 months, preferably at least 12 months, more preferably at least 18 months.

[0084] Polymorph II

[0085] In one embodiment, the present invention also relates to Polymorph II of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid.

[0086] Polymorph II is characterized in that its XRPD pattern comprises peaks at 2θ values of 10.8°, 12.1°, 12.4° and 22.3°.

[0087] The 2θ values provided herein are for measurements conducted at room temperature under the conditions detailed in Example 2. In one embodiment, an X-ray wavelength is used to measure the peaks of the XRPD pattern. In one embodiment, the error range of the provided angular values is ±0.2°, preferably ±0.1°.

[0088] In one embodiment, the XRPD pattern of Polymorph II comprises peaks at 2θ values of 10.8°, 12.1°, 12.4°, 15.3°, 16.2°, 18.5°, 19.5°, 20.7°, 21.5° and 22.3°.

[0089] In one embodiment, the XRPD pattern of Polymorph II comprises peaks with 2θ values similar to those shown in Table 2.

[0090] Table 2. XRPD pattern of Polymorph II and relative intensities of the reflection peaks (2θ).

[0091] 2θ angle (°) Relative intensity (%) 10.4 19.3 10.8 100 12.1 68.6 12.4 29.9 15.3 11.8 16.2 21.8 18.5 17.7 18.7 13.6 19.5 14.5 19.7 11.3 20.7 24.7 21.5 16.7 22.3 47.5 24.4 12.4 24.8 13.1

[0092] In one embodiment, Polymorph II has an XRPD pattern substantially as Figure 5 shown.

[0093] In one embodiment, the DSC thermogram of Polymorph II shows a first endothermic value with a peak temperature of about 110 °C, starting at about 100 °C; an exothermic peak with a peak temperature of about 149 °C, starting at about 134 °C; and a second endothermic peak with a peak temperature of about 181 °C, starting at about 179 °C.

[0094] In one embodiment, Polymorph II has a DSC thermogram substantially as Figure 6 shown.

[0095] In one embodiment, Polymorph II has a TGA thermogram substantially as Figure 7 shown, which exhibits no significant weight loss at temperatures below 220 °C.

[0096] It has been demonstrated that Polymorph II is advantageously non-hygroscopic. In particular, a water adsorption / desorption curve has been established for Polymorph II, as Figure 8 reported in the DVS isotherm plot of and as detailed in Example 6.

[0097] Manufacturing method

[0098] Polymorph I and Polymorph II can be prepared by various methods available to those skilled in the art, including the methods described below and in the examples.

[0099] Accordingly, the present invention relates to a method for preparing Polymorph I of the present invention, which comprises crystallizing (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid from an ethanol / water mixture.

[0100] In one embodiment, the method for preparing Polymorph I comprises crystallizing (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (Compound (1)) from an ethanol / water mixture in a volume-to-weight ratio of 1 to 20, preferably, based on the volume of the solvent mixture and the weight of the compound, at a temperature of up to 80 °C, and in a ratio of ethanol / water of 95 / 5 to 5 / 95.

[0101] "Volume-to-weight ratio" refers to the ratio of the volume of the solvent involved in the method (e.g., expressed in mL or L) to the weight of the compound (e.g., expressed in g or kg, respectively).

[0102] In one embodiment, the method for preparing Polymorph I comprises the following steps:

[0103] 1) Dissolving (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (Compound (1)) in an ethanol / water mixture at a temperature of up to 80 °C, preferably 40 °C to 60 °C, more preferably 45 °C to 55 °C, wherein the ethanol / water ratio is from 100 / 0 to 5 / 95;

[0104] 2) If necessary, adding water to bring the ethanol / water ratio to 95 / 5 to 5 / 95, wherein the water is preferably heated at the same temperature as the ethanol used in step 1); and

[0105] 3) Cool the mixture, preferably to a temperature of from 0 °C to 10 °C, more preferably from 3 °C to 5 °C, and maintain this temperature for a period of time suitable for recovering polymorph I.

[0106] In one embodiment, in step 1), compound (1) is directly dissolved in an ethanol / water mixture having the ethanol / water ratio required for crystallization (i.e., from 95 / 5 to 5 / 95). In this case, step 2) is not present.

[0107] In another embodiment, in step 1), compound (1) is dissolved in an ethanol / water mixture containing more ethanol than the ethanol / water ratio required for crystallization. In this case, water is added according to step 2) to achieve the ethanol / water ratio of 95 / 5 to 5 / 95 required for crystallization. In a specific embodiment, in step 1), compound (1) is dissolved in ethanol, and in step 2), water is added to bring the ethanol / water ratio to 95 / 5 to 5 / 95. In one embodiment, when compound (1) is dissolved in ethanol in step 1), it can be filtered and rinsed with a small amount of ethanol heated at the working temperature, and then water is added to the resulting ethanol filtrate according to step 2).

[0108] In one embodiment, the ethanol / water ratio (volume ratio) for crystallizing compound (1) into polymorph I is from 95 / 5 to 5 / 95, preferably from 95 / 5 to 50 / 50, more preferably from 95 / 5 to 65 / 35. In one embodiment, the ethanol / water ratio for crystallizing compound (1) into polymorph I is 95 / 5 volume ratio. In another embodiment, the ethanol / water ratio for crystallizing compound (1) into polymorph I is 75 / 25 volume ratio. In another embodiment, the ethanol / water ratio for crystallizing compound (1) into polymorph I is 70 / 30 volume ratio.

[0109] In one embodiment, the volume-to-weight ratio of the ethanol / water mixture used for crystallizing compound (1) into polymorph I to compound (1) is from 1 to 20, preferably from 3 to 7. In one embodiment, the volume-to-weight ratio of the ethanol / water mixture used for crystallizing compound (1) into polymorph I to compound (1) is equal to 3. In another embodiment, the volume-to-weight ratio of the ethanol / water mixture used for crystallizing compound (1) into polymorph I to compound (1) is equal to 9. In another embodiment, the volume-to-weight ratio of the ethanol / water mixture used for crystallizing compound (1) into polymorph I to compound (1) is equal to 6.

[0110] In one embodiment, the cooling in step 3) can be achieved at different time periods and different temperature ranges. For example, the mixture can be cooled to a temperature of 3°C to 5°C for about 6 hours and then maintained at that temperature for about 12 hours. Alternatively, the mixture can be rapidly cooled to 37°C, then held isothermal for 3 hours, and then further cooled to about 5°C within 16 hours. Depending on the cooling parameters, different recovery yields and purities can be achieved. Those skilled in the art will be able to determine suitable cooling conditions.

[0111] The crystals obtained in step 3) can be separated by filtration and dried under reduced pressure as needed.

[0112] The present invention also relates to a crystalline form of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid obtainable by a method comprising the following steps:

[0113] 1) Dissolving (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (Compound (1)) in an ethanol / water mixture at a temperature of up to 80°C, preferably 40°C to 60°C, more preferably 45°C to 55°C, wherein the ethanol / water ratio is from 100 / 0 to 5 / 95;

[0114] 2) If necessary, adding water to bring the ethanol / water ratio to 95 / 5 to 5 / 95, wherein the water is preferably heated at the same temperature as the ethanol used in step 1); and

[0115] 3) Cooling the mixture, preferably to a temperature of 0°C to 10°C, more preferably 3°C to 5°C, and maintaining that temperature for a period suitable for recovering the crystals.

[0116] The present invention also relates to a method for preparing crystalline form II of the present invention, which comprises crystallizing (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid from acetonitrile.

[0117] In one embodiment, the method for preparing crystalline form II comprises crystallizing (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (Compound (1)) from acetonitrile at a temperature ranging from room temperature up to 80°C at a volume-to-weight ratio of acetonitrile to Compound (1) of 2 to 3.

[0118] In one embodiment, the method for preparing crystalline form II comprises the following steps:

[0119] 1) Dissolve (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (Compound (1)) in acetonitrile at a temperature of at most 80 °C; and

[0120] 2) Cool the mixture of step 1), preferably to room temperature, and maintain this temperature during a period suitable for recovering polymorph II.

[0121] In one embodiment, the volume-to-weight ratio of acetonitrile to Compound (1) used in the method for preparing polymorph II is 2 to 3, preferably 2.2 to 2.3.

[0122] In one embodiment, the cooling in step 2) can be achieved at different time periods and within different temperature ranges. For example, the mixture of step 1) can be cooled to room temperature and then maintained at this temperature for about 1 hour. Depending on the cooling parameters, different recovery yields and purities can be achieved. Those skilled in the art can determine suitable cooling conditions.

[0123] The crystals obtained in step 2) can be separated by filtration and dried under reduced pressure as needed.

[0124] The present invention also relates to a polymorph of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid obtainable by a method comprising the following steps:

[0125] 1) Dissolve (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (Compound (1)) in acetonitrile at a temperature of at most 80 °C; and

[0126] 2) Cool the mixture of step 1), preferably to room temperature, and maintain this temperature during a period suitable for recovering the crystals.

[0127] Seeds can be added to any crystallization mixture to facilitate crystallization.

[0128] Polymorph I or polymorph II can be directly prepared from the reaction medium of the final process for preparing Compound (1). Alternatively, after one or more preliminary crystallization steps, recrystallization can be carried out according to the above method to obtain polymorph I or polymorph II.

[0129] Due to their properties, the above methods for preparing polymorph I or polymorph II can be advantageously applied downstream of any synthesis method of Compound (1) described in the prior art.

[0130] In particular, the methods for preparing polymorph I and polymorph II can be applied downstream of the method for preparing compound (1) reported in WO2021 / 250174.

[0131] Thus, in one embodiment, there is provided a method for preparing polymorph I, which comprises:

[0132] a) A step of hydrochloride formation and epimer separation, which comprises crystallizing a mixture of epimers (6) and (6') in the presence of hydrochloric acid in a solvent selected from alcohol solvents (such as isopropanol, methanol and their mixtures), isopropyl acetate and their mixtures:

[0133]

[0134] wherein the epimer ratio (6):(6') is at least 4:1;

[0135] to obtain methyl (2S,5R)-5-(2-chlorophenyl)pyrrolidine-2-carboxylate hydrochloride (6.HCl):

[0136]

[0137] b) Forming methyl ester (9) from the compound (6.HCl) obtained in step a),

[0138]

[0139] b1) By acylating the compound (6.HCl) with the compound (7) in a mixture of toluene and water as the solvent in the presence of potassium carbonate:

[0140]

[0141] Or

[0142] b2) By coupling the compound (6.HCl) with the compound (8) in the presence of a base and an acid activator:

[0143]

[0144] c) Saponifying the methyl ester (9) in a mixture of toluene and water as the solvent in the presence of sodium hydroxide and tetrabutylammonium bromide (TBAB) to obtain the acid compound (1); and

[0145] d) Performing the method for preparing polymorph I of the present invention as detailed above.

[0146] In one embodiment, an intermediate purification step c can be carried out between step c) and step d) in the above method 1) so as to further purify compound (1) before crystallization in Form I.

[0147] In one embodiment, a method for preparing Form II is also provided, which comprises:

[0148] the above steps a), b) and c); and

[0149] d’) performing the method for preparing Form II of the present invention as detailed above.

[0150] In one embodiment, an intermediate purification step c 1 ) can be carried out between step c) and step d’) in the above method so as to further purify compound (1) before crystallization in Form II.

[0151] Pharmaceutical composition

[0152] The present invention also provides a pharmaceutical composition comprising Form I and / or Form II according to the present invention and at least one pharmaceutically acceptable carrier.

[0153] In one embodiment, the present invention provides a pharmaceutical composition comprising Form I according to the present invention and at least one pharmaceutically acceptable carrier. In another embodiment, the present invention provides a pharmaceutical composition comprising Form II according to the present invention and at least one pharmaceutically acceptable carrier. In another embodiment, the present invention provides a pharmaceutical composition comprising a mixture of Form I and Form II according to the present invention and at least one pharmaceutically acceptable carrier.

[0154] In one embodiment, the pharmaceutical composition of the present invention comprises Form I and / or Form II, at least one pharmaceutically acceptable carrier and optionally one or more additional pharmaceutically active agents.

[0155] By way of non-limiting example, the pharmaceutical composition of the present invention can be in a form suitable for oral administration, parenteral administration (e.g., by intravenous, intramuscular or subcutaneous injection or intravenous infusion), topical administration (including ophthalmic administration), administration by inhalation, skin patch, implant, suppository, etc. These suitable forms of administration (which can be solid, semi-solid or liquid depending on the mode of administration) and the methods and carriers, diluents and excipients used in their preparation will be clear to those skilled in the art; reference is made to the latest edition of Remington’s Pharmaceutical Sciences.

[0156] Some preferred but non-limiting examples of such pharmaceutical preparations include tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, creams, lotions, soft and hard gelatin capsules, suppositories, drops, sterile injectable solutions and sterile packaged powders (which are usually reconstituted before use), which can be formulated together with carriers, excipients and diluents suitable for such preparations, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, polyethylene glycol, cellulose, (sterile) water, methylcellulose, methyl and propyl hydroxybenzoates, talc, magnesium stearate, edible oils, vegetable oils and mineral oils or suitable mixtures thereof. The preparations may also optionally contain other substances commonly used in pharmaceutical preparations, such as lubricants, wetting agents, emulsifying and suspending agents, dispersing agents, disintegrants, swelling agents, fillers, preservatives, sweeteners, flavoring agents, flow regulators, demolding agents, etc. The compositions can also be formulated to provide rapid, sustained or delayed release of the active compounds contained therein.

[0157] In one embodiment, the pharmaceutical composition of the present invention is in a form suitable for oral administration, preferably selected from tablets, pills, soft and hard gelatin capsules, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions and syrups.

[0158] The pharmaceutical composition of the present invention is preferably in the form of unit dosage forms and can be suitably packaged in, for example, boxes, blisters, vials, bottles, sachets, ampoules or any other suitable single-dose or multi-dose drug-carrying devices or containers (which can be suitably labeled); optionally with one or more leaflets containing product information and / or instructions for use.

[0159] Generally, such unit doses will contain 0.05 to 1000 mg, usually 1 to 500 mg of crystalline form I and / or crystalline form II according to the present invention, such as about 10, 25, 50, 100, 200, 300 or 400 mg per unit dose.

[0160] Typically, depending on the condition to be prevented or treated and the route of administration, crystalline form I and / or crystalline form II can usually be administered in an amount of 0.01 to 100 mg per kilogram of patient body weight per day, more commonly 0.1 to 50 mg, such as 1 to 25 mg, such as about 0.5, 1, 5, 10, 15, 20 or 25 mg, which can be administered as a single daily dose, divided into one or more daily doses or administered substantially continuously, such as using infusion.

[0161] Medical Use

[0162] The compound (1), (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid, is an agonist of FFAR2 and can be used for the treatment of diseases related to or mediated by FFAR2, especially inflammatory diseases and cancers.

[0163] Therefore, the present invention provides the use of polymorphic form I and / or polymorphic form II as a medicament.

[0164] The present invention also provides the use of polymorphic form I and / or polymorphic form II for the treatment of inflammation.

[0165] In one embodiment, polymorphic form I and / or polymorphic form II are used for the treatment of inflammation.

[0166] In one embodiment, polymorphic form I and / or polymorphic form II are used for the treatment of inflammatory diseases.

[0167] In one embodiment, polymorphic form I and / or polymorphic form II are used for delaying the onset of inflammatory diseases in a patient.

[0168] In one embodiment, polymorph I and / or polymorph II are used for treating diseases selected from the following: rheumatoid arthritis; inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which results in bacterial infections (e.g., Clostridium difficile infection, pneumococcal pneumonia, etc.); Paget's disease; osteoporosis; multiple myeloma; uveitis; acute myeloid leukemia; chronic myeloid leukemia; pancreatic beta cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritic conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis, including hepatitis B virus infection; asthma; muscle degeneration; cachexia, such as cachexia secondary to infection or malignancy, cachexia secondary to acquired immunodeficiency syndrome; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; autoimmune brain diseases, such as encephalitis and encephalomyelitis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; Gram-negative sepsis; fever and myalgia caused by infection (e.g., influenza); heartburn; inflammatory conditions triggered by the release of anorectic gut hormones (e.g., PYY, GLP-1), such as inflammatory conditions observed in hyperphagia-related disorders, obesity, type 2 diabetes, etc.; microbiome-related neurological and mood disorders, including autism spectrum disorder, schizophrenia, depression, major depressive disorder, and neurodegenerative diseases characterized by neuroinflammation, including Alzheimer's disease and Parkinson's disease.

[0169] In one embodiment, Form I and / or Form II are used for treating diseases selected from the following: rheumatoid arthritis; inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which leads to bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia, etc.); Paget's disease; osteoporosis; multiple myeloma; uveitis; acute and chronic myeloid leukemia; pancreatic beta cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritic conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis, including hepatitis B virus infection; asthma; muscle degeneration; cachexia, such as cachexia secondary to infection or malignancy, cachexia secondary to acquired immunodeficiency syndrome; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; gram-negative sepsis; fever and myalgia caused by infection (such as influenza); heartburn; inflammatory conditions triggered by the release of anorectic gut hormones (such as PYY, GLP-1), such as the inflammatory conditions observed in hyperphagia-related disorders, obesity, type 2 diabetes, etc.; microbiome-related neurological and mood disorders, including autism spectrum disorder, schizophrenia, depression, major depressive disorder and neurodegenerative diseases characterized by neuroinflammation, including Alzheimer's disease and Parkinson's disease.

[0170] In one embodiment, Form I and / or Form II are used for treating diseases selected from the following: inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, colitis, collagenous colitis, lymphocytic colitis, immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), pouchitis, celiac disease, irritable bowel syndrome, gut microbiota dysbiosis, type 1 diabetes, multiple sclerosis, autoimmune brain diseases, such as encephalitis and encephalomyelitis.

[0171] In one embodiment, Form I and / or Form II are used for treating diseases selected from the following: inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which leads to bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia infection, etc.).

[0172] The present invention also relates to a method for treating inflammation in a patient in need thereof, which comprises administering to the patient a therapeutically effective amount of Form I and / or Form II of the present invention.

[0173] In one embodiment, the present invention provides a method for treating an inflammatory disease in a patient in need thereof, which comprises administering to the patient a therapeutically effective amount of Form I and / or Form II of the present invention.

[0174] In one embodiment, the present invention provides a method for delaying the onset of an inflammatory disease in a patient in need thereof, which comprises administering to the patient a therapeutically effective amount of Form I and / or Form II of the present invention.

[0175] In one embodiment, the present invention provides a method for treating a disease in a patient in need thereof, said disease being selected from: rheumatoid arthritis; inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which results in bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia, etc.); Paget's disease; osteoporosis; multiple myeloma; uveitis; acute myeloid leukemia; chronic myeloid leukemia; pancreatic beta cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritic conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis, including hepatitis B virus infection; asthma; muscle degeneration; cachexia, such as cachexia secondary to infection or malignancy, cachexia secondary to acquired immunodeficiency syndrome; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; autoimmune brain diseases, such as encephalitis and encephalomyelitis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; gram-negative sepsis; fever and myalgia caused by infection (such as influenza); heartburn; inflammatory conditions triggered by the release of anorectic gut hormones (such as PYY, GLP-1), such as the inflammatory conditions observed in hyperphagia-related disorders, obesity, type 2 diabetes, etc.; microbiome-related neuro and mood disorders, including autism spectrum disorder, schizophrenia, depression, major depressive disorder and neurodegenerative diseases characterized by neuroinflammation, including Alzheimer's disease and Parkinson's disease; the method comprising administering to the patient a therapeutically effective amount of polymorph I and / or polymorph II of the present invention.

[0176] In one embodiment, the present invention provides a method of treating a disease in a patient in need thereof, said disease being selected from: rheumatoid arthritis; inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which leads to bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia in the lungs, etc.); Paget's disease; osteoporosis; multiple myeloma; uveitis; acute and chronic myeloid leukemia; pancreatic beta cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritis conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis, including hepatitis B virus infection; asthma; muscle degeneration; cachexia, such as cachexia secondary to infection or malignancy, cachexia secondary to acquired immunodeficiency syndrome; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; Gram-negative sepsis; fever and myalgia caused by infection (such as influenza); heartburn; inflammatory conditions triggered by the release of anorectic gut hormones (such as PYY, GLP-1), such as the inflammatory conditions observed in hyperphagia-related disorders, obesity, type 2 diabetes, etc.; microbiome-related neurological and mood disorders, including autism spectrum disorder, schizophrenia, depression, major depressive disorder and neurodegenerative diseases characterized by neuroinflammation, including Alzheimer's disease and Parkinson's disease; the method comprising administering to the patient a therapeutically effective amount of polymorph I and / or polymorph II of the present invention.

[0177] In one embodiment, the present invention provides a method for treating a disease in a patient in need thereof, said disease being selected from: inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, colitis, collagenous colitis, lymphocytic colitis, immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), pouchitis, celiac disease, irritable bowel syndrome, gut microbiota dysbiosis, type 1 diabetes, multiple sclerosis, autoimmune brain diseases such as encephalitis and encephalomyelitis; the method comprising administering to the patient a therapeutically effective amount of polymorph I and / or polymorph II of the present invention.

[0178] In one embodiment, the present invention provides a method for treating a disease in a patient in need thereof, said disease being selected from: inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis which results in bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia infection in the lungs, etc.); the method comprising administering to the patient a therapeutically effective amount of polymorph I and / or polymorph II of the present invention.

[0179] The present invention also relates to the use of polymorph I and / or polymorph II of the present invention in the preparation of a medicament for treating inflammation.

[0180] In one embodiment, the present invention provides the use of polymorph I and / or polymorph II of the present invention in the manufacture of a medicament for treating inflammatory diseases.

[0181] In one embodiment, the present invention provides the use of polymorph I and / or polymorph II of the present invention in the manufacture of a medicament for delaying the onset of an inflammatory disease in a patient.

[0182] In one embodiment, the present invention provides the use of polymorph I and / or polymorph II of the present invention in the preparation of a medicament for treating a disease selected from the group consisting of: rheumatoid arthritis; inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which leads to bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia in the lungs, etc.); Paget's disease; osteoporosis; multiple myeloma; uveitis; acute myeloid leukemia; chronic myeloid leukemia; pancreatic beta cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritic conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis, including hepatitis B virus infection; asthma; muscle degeneration; cachexia, such as cachexia secondary to infection or malignancy, cachexia secondary to acquired immunodeficiency syndrome; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; autoimmune brain diseases, such as encephalitis and encephalomyelitis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; Gram-negative sepsis; fever and myalgia caused by infection (such as influenza); heartburn; inflammatory conditions triggered by the release of anorectic gut hormones (such as PYY, GLP-1), such as the inflammatory conditions observed in hyperphagia-related disorders, obesity, type 2 diabetes, etc.; microbiome-related neuro and mood disorders, including autism spectrum disorder, schizophrenia, depression, major depressive disorder, and neurodegenerative diseases characterized by neuroinflammation, including Alzheimer's disease and Parkinson's disease.

[0183] In one embodiment, the present invention provides the use of Form I and / or Form II of the present invention in the manufacture of a medicament for treating a disease selected from the group consisting of: rheumatoid arthritis; inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which leads to bacterial infections (such as Clostridium difficile infection, pneumococcal pneumonia, etc.); Paget's disease; osteoporosis; multiple myeloma; uveitis; acute and chronic myelogenous leukemia; pancreatic beta cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritic conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis, including hepatitis B virus infection; asthma; muscle degeneration; cachexia, such as cachexia secondary to infection or malignancy, cachexia secondary to acquired immunodeficiency syndrome; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; gram-negative sepsis; fever and myalgia caused by infection (such as influenza); heartburn; inflammatory conditions triggered by the release of anorectic gut hormones (such as PYY, GLP-1), such as the inflammatory conditions observed in hyperphagia-related disorders, obesity, type 2 diabetes, etc.; microbiome-related neurological and mood disorders, including autism spectrum disorder, schizophrenia, depression, major depressive disorder, and neurodegenerative diseases characterized by neuroinflammation, including Alzheimer's disease and Parkinson's disease.

[0184] In one embodiment, the present invention provides the use of Form I and / or Form II of the present invention in the manufacture of a medicament for treating a disease selected from the group consisting of: inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, colitis, collagenous colitis, lymphocytic colitis, immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), immune-mediated colitis (IMC) (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), pouchitis, celiac disease, irritable bowel syndrome, gut microbiota dysbiosis, type 1 diabetes, multiple sclerosis, autoimmune brain diseases, such as encephalitis and encephalomyelitis.

[0185] In one embodiment, the present invention provides the use of Form I and / or Form II of the present invention in the manufacture of a medicament for treating a disease selected from the group consisting of: inflammatory bowel disease (IBD), including but not limited to Crohn's disease and ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis (including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1), pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis, including antibiotic-induced dysbiosis, which leads to bacterial infections (such as Clostridioides difficile infection, pneumococcal pneumonia, etc.).

[0186] The present invention also provides the use of Form I and / or Form II for treating cancer.

[0187] In one embodiment, Form I and / or Form II is used for treating cancer.

[0188] The present invention also relates to a method of treating cancer in a patient in need thereof, which comprises administering to the patient a therapeutically effective amount of Form I and / or Form II of the present invention.

[0189] The present invention also relates to the use of Form I and / or Form II of the present invention in the preparation of a medicament for treating cancer.

[0190] In one embodiment, Form I and / or Form II is used for delaying the onset of cancer in a patient.

[0191] In one embodiment, the present invention provides a method of delaying the onset of cancer in a patient in need thereof, which comprises administering to the patient a therapeutically effective amount of Form I and / or Form II of the present invention.

[0192] In one embodiment, the present invention provides the use of Form I and / or Form II of the present invention in the manufacture of a medicament for delaying the onset of cancer in a patient.

[0193] Non-limiting examples of cancers include lung cancer, non-small cell lung cancer, small cell lung cancer, breast cancer, prostate cancer, ovarian cancer, endometrial cancer, vaginal cancer, testicular cancer, cervical cancer, bladder cancer, head and neck cancer, kidney cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, thyroid cancer, gastrointestinal cancer, colorectal cancer, gastric cancer, colon cancer, liver cancer, leukemia, lymphoma, skin cancer, melanoma, multiple myeloma, glioma, glioblastoma, mesothelioma, retinoblastoma, sarcoma, Ewing's sarcoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, bone cancer, and gastric cardia cancer.

[0194] In certain embodiments, Polymorph I and / or Polymorph II are used in combination with additional agents. For example, the additional agent can be a second anti-cancer therapy such as chemotherapy, immunotherapy, cell therapy, surgery, transplantation, and / or any anti-cancer agent currently in clinical use or in clinical trials.

[0195] In the context of the present invention, the term "combination" preferably means the presence of a combination of Polymorph I and / or Polymorph II with an additional agent. Thus, the combination can exist as a composition that contains all the components in the same mixture (e.g., a pharmaceutical composition), or can exist as a multi-part kit, where the different components form different parts of such a multi-part kit. The administration of Polymorph I and / or Polymorph II and the additional agent can be carried out simultaneously, or can be staggered in time with similar or different administration schedules (i.e., the number of administrations of each component is similar or different), at the same administration site or different administration sites, using similar or different dosage forms.

[0196] Brief Description of the Drawings

[0197] Figure 1 is the X-ray powder diffraction (XRPD) pattern of Polymorph I.

[0198] Figure 2 is the differential scanning calorimetry (DSC) thermogram of Polymorph I.

[0199] Figure 3 is the thermogravimetric analysis (TGA) thermogram of Polymorph I.

[0200] Figure 4 is the dynamic vapor sorption (DVS) isotherm of Polymorph I.

[0201] Figure 5 is the XRPD pattern of Polymorph II.

[0202] Figure 6 is the DSC thermogram of Polymorph II.

[0203] Figure 7 is the TGA thermogram of Polymorph II.

[0204] Figure 8 is the DVS isotherm of Polymorph II. Examples

[0205] The present invention is further illustrated by the following examples.

[0206] Example 1: Preparation of Polymorph I and Polymorph II

[0207] The starting material (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid (referred to herein as compound (1)) was obtained as described in WO2021 / 250174.

[0208] Crystal form I

[0209] Crystallization of compound (1) in crystal form I was achieved from a 6-fold volume of a 70:30 ethanol:water mixture.

[0210] Compound (1) was dissolved in 3.7-fold volume of ethanol at 48 - 52 °C, filtered, and rinsed with 0.5-fold volume of ethanol (heated to 48 - 52 °C).

[0211] 1.8-fold volume of deionized water was heated to 48 - 52 °C, filtered, and added to the ethanol filtrate. The mixture was cooled to 3 - 5 °C over 6 hours and then held at that temperature for 12 hours.

[0212] The wet product was filtered, sieved if necessary, and dried at 40 °C. It was identified as crystal form I, as detailed below.

[0213] Crystal form II

[0214] Compound (1) (3.57 g, 7.7 mmol) was dissolved in acetonitrile (8 ml) and stirred at 80 °C. After 2 - 3 minutes, complete dissolution was obtained and crystallization began. The mixture was stirred at room temperature for 1 hour.

[0215] The crystals were filtered and dried under reduced pressure (10 mbar) for 20 hours to give 2.45 g of white crystals, which were identified as crystal form II, as detailed below.

[0216] Example 2: X-ray powder diffraction (XRPD)

[0217] X-ray powder diffraction (XRPD) analysis was performed on a PANalytical X’Pert Pro diffractometer in transmission mode (40 kV, 40 mA, Cu Kα 1 ). The analysis was carried out in the 2θ value range from 2° to 50° (continuous scan mode, scan rate 0.04 degrees per second).

[0218] The XRPD pattern of crystal form I is reported in Figure 1 The selected diffraction peaks are shown in Table 1 provided in the detailed description above.

[0219] The XRPD pattern of crystal form II is reported in Figure 5 The selected diffraction peaks are shown in Table 2 provided in the detailed description above.

[0220] Those skilled in the art will understand that the obtained XRPD patterns may have measurement errors, which depend on the measurement conditions employed. In particular, it is known that the intensities in the XRPD patterns may fluctuate according to the measurement conditions used. It should also be understood that the relative intensities may also vary according to the experimental conditions, and thus, the exact order of the intensities should not be considered. In addition, the measurement error of the diffraction angle of a conventional XRPD pattern is generally about 5% or less, and such a degree of measurement error should be considered in relation to the aforementioned diffraction angle.

[0221] Therefore, it should be understood that Polymorph I and Polymorph II of the present invention are not limited to crystal structures providing XRPD patterns identical to those shown in Figure 1 and Figure 5 Any crystal structure providing an XRPD pattern substantially identical to the XRPD patterns disclosed in Figure 1 or Figure 5 falls within the scope of the present invention. The ability to determine the substantial identity of XRPD patterns is within the capabilities of those of ordinary skill in the art.

[0222] Example 3: Single Crystal X-ray Analysis

[0223] Single crystals of Polymorph I were obtained and studied by X-ray diffraction.

[0224] Data collection was carried out on a Rigaku Ru200 rotating anode (Mo, ). The X-rays were monochromatized with a Zr filter and the data were recorded on a MAR345 image plate. A 0.18 x 0.14 x 0.06 mm crystal was selected for the experiment and mounted in a nylon loop on the diffractometer. During the measurement, the crystal was cooled to 120 K in gaseous nitrogen. A total of 220 images were collected with a phi increment of 3°. The MAR program was used to collect the diffraction data and the AUTOMAR suite was used to process the data. No absorption correction was applied. The structure was solved by the direct method (SHELXS) and refined against F2 by full least-squares refinement using (SHELXL-97). The non-hydrogen atoms were refined anisotropically and the hydrogen atoms were placed at calculated positions and refined riding on the parent atoms. The absolute configuration was calculated based on the anomalous dispersion effect, and the final Flack parameter was 0.07(12), the Hooft parameter was 0.08(3), and the Friedel pair coverage was 100%.

[0225] Polymorph I crystallizes in an orthorhombic unit cell structure with unit cell parameters equal to:

[0226] Unit cell dimensions:

[0227]

[0228] α, β, γ = 90°

[0229] Space group: P2 1 2 1 2 1

[0230] Number of molecules per unit cell: 4

[0231] Unit cell volume:

[0232] Density (calculated): 1.19 g / cm 3

[0233] Mu(MoKa)[ / mm] = 0.186

[0234] F(000) = 912

[0235] Example 4: Differential Scanning Calorimetry (DSC)

[0236] Differential scanning calorimetry (DSC) analysis was carried out on a Mettler Toledo DSC3+ instrument. The sample (a few milligrams) was placed in a 40 μL aluminum sample pan and crimped with a perforated lid. The analysis was performed under a nitrogen purge (50 mL / min) with a temperature scan from 20 °C to 300 °C at 10 °C / min.

[0237] As Figure 2 shown, the DSC thermogram of polymorph I shows an endothermic peak with a peak temperature of approximately 183 °C and a start temperature of approximately 180 °C. It was demonstrated by its DSC analysis that polymorph I is a non-solvated polymorph.

[0238] As Figure 6 shown, the DSC thermogram of polymorph II shows a first endothermic peak signal that starts at approximately 100 °C and has a peak temperature of approximately 110 °C, corresponding to the melting of the sample. Then, a recrystallization-melting phenomenon was observed, which has an exothermic peak signal that starts at approximately 134 °C and has a peak temperature of approximately 149 °C; and an endothermic peak signal that starts at approximately 179 °C and has a peak temperature of approximately 181 °C. It was demonstrated by its DSC analysis that polymorph II is a non-solvated polymorph.

[0239] Those skilled in the art will understand that in DSC measurements, there is a certain degree of variation in the actually measured start temperature and peak temperature, which depends on the heating rate, crystal shape and purity, and other measurement parameters.

[0240] Therefore, it should be understood that polymorph I and polymorph II of the present invention are not limited to providing the same as Figure 2 and Figure 6The crystal structure of a DSC thermogram that is identical to the DSC thermogram shown. Any crystal structure that provides a DSC thermogram substantially the same as the DSC thermogram disclosed in Figure 2 or Figure 6 falls within the scope of the present invention. The ability to determine the basic identity of a DSC thermogram is within the capabilities of those of ordinary skill in the art.

[0241] Example 5: Thermogravimetric Analysis (TGA)

[0242] Thermogravimetric analysis (TGA) experiments were carried out on a Mettler Toledo TGA / DSC3+ instrument. Samples (about 10 - 30 mg) were placed in 100 μL aluminum sample pans and crimped with a perforated lid. The analysis was carried out under a nitrogen purge (50 mL / min) with a temperature scan between 25 °C and 300 °C at a heating rate of 10 °C / min.

[0243] As Figure 3 shown, the TGA analysis of polymorph I did not show any significant weight loss at temperatures below 220 °C: a weight loss of -0.14% was measured in the temperature range of 10 °C - 200 °C. Above this temperature, the observed weight loss may be due to evaporation and / or degradation, and this event did not end at the end of the analysis.

[0244] As Figure 7 shown, the TGA analysis of polymorph II did not show any significant weight loss at temperatures below 220 °C: a weight loss of -0.2% was measured in the temperature range of 10 °C - 200 °C. Above this temperature, the observed weight loss may be due to evaporation and / or degradation of the sample.

[0245] Example 6: Hygroscopicity Study – Dynamic Vapor Sorption (DVS)

[0246] The previously described solid forms of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid, namely the amorphous free acid and the corresponding sodium salt (both disclosed in WO2021 / 250174), are hygroscopic.

[0247] In particular, the amorphous free acid form showed an 8 wt% increase after only 14 hours at 70 - 75% relative humidity at room temperature, and a 3 wt% increase after 2 days at 40% relative humidity.

[0248] The Na salt showed a 20 wt% increase after only 14 hours at 22 °C and 70% relative humidity, and a 7 wt% increase after 14 hours at 40% relative humidity.

[0249] Therefore, it is crucial to determine the hygroscopicity of polymorph I and polymorph II.

[0250] It has been demonstrated that after 10 days at 25 °C and 80% relative humidity, polymorph I showed only a 0.09 wt% increase.

[0251] The hygroscopicity of polymorphs I and II was also evaluated by dynamic vapor sorption (DVS).

[0252] Analysis was performed on a surface measurement system DVS Intrinsic. The sample was placed in an open aluminum pan. Analysis was carried out at 25 °C. The relative humidity (RH) was scanned between 0% RH and 95% RH in 10% RH steps (40–0–95–0–95). The stability criterion was a mass change below 0.002% within a 5-minute time window, with a minimum step time of 10 minutes and a maximum step time of 100 minutes.

[0253] As Figure 4 shown, when exposed to high relative humidity values, the DVS analysis did not show any significant weight increase for polymorph I. During the first desorption phase (40% RH to 0% RH), a weight loss of -0.1% was observed. During the first adsorption phase (0% RH to 95% RH), a small amount of water uptake was observed, mainly at high relative humidity values: at the end of the 70% RH step, the weight increase was +0.1%, and it reached a maximum of +0.7% at 95% RH. During the second desorption phase (95% RH to 0% RH), the sample lost all the water gained during the first adsorption phase and returned to the value observed at the end of the first desorption phase. The second adsorption phase (0% RH to 95% RH) was similar to the first adsorption phase.

[0254] As Figure 8 shown, when exposed to high relative humidity values, the DVS analysis did not show any significant weight increase for polymorph II. During the first desorption phase (40% RH to 0% RH), a weight loss of -0.1% was observed. During the first adsorption phase (0% RH to 95% RH), a small amount of water uptake was observed, mainly at high relative humidity values: at the end of the 70% RH step, the weight increase was +0.1%, and it reached a maximum of +0.4% at 95% RH. During the second desorption phase (95% RH to 0% RH), the sample lost all the water gained during the first adsorption phase and returned to the value observed at the end of the first desorption phase. The second adsorption phase (0% RH to 95% RH) was similar to the first adsorption phase.

[0255] At the end of the analysis, the recovered sample was analyzed by X-ray diffraction. No phase transformation was detected: the diffraction pattern corresponded to that of each starting material.

[0256] Conclusion: It has been demonstrated that both polymorphic forms I and II are non-hygroscopic, which is contrary to what was observed for the amorphous form and the Na salt of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid.

[0257] Example 7: Stability Study

[0258] The relative stabilities of polymorphic forms I and II were evaluated by competitive slurry tests and cross-seeding tests. Analysis demonstrated that polymorphic form I is more stable than polymorphic form II.

[0259] Competitive slurry test in isopropanol

[0260] Competitive slurry tests between polymorphic forms I and II of compound (1) were carried out in isopropanol at 10 °C, 20 °C, 25 °C, 30 °C and 40 °C.

[0261] Method. First, a suspension of polymorphic form II of compound (1) was prepared in isopropanol and equilibrated for 30 minutes at each temperature to be studied. Then, approximately 800 μL of the saturated solution was sampled, filtered through a 0.2 μm HPTFE filter, and added to a vial equipped with a magnetic stir bar containing 50 mg of polymorphic form I and 50 mg of polymorphic form II. After 10 seconds, the first sample of the solid phase was taken and analyzed by XRPD to identify the solid phase present. Additional samplings were also carried out until only 1 solid phase remained.

[0262] Results. The results of XRPD analysis at each temperature were compared with the diffraction patterns of polymorphic forms I and II. Since the first sampling (i.e., after 10 seconds), polymorphic form II was no longer visible in the suspension. The results indicate that polymorphic form I is more stable than polymorphic form II in isopropanol within the tested temperature range.

[0263] Competitive slurry test in n-heptane

[0264] Competitive slurry tests between polymorphic forms I and II of compound (1) were carried out in n-heptane at 10 °C, 20 °C, 25 °C, 30 °C and 40 °C.

[0265] Method. Approximately 1 mL of n-heptane was added to a vial equipped with a magnetic stir bar containing 40 mg of polymorphic form I and 40 mg of polymorphic form II. After 10 seconds, the first sample of the solid phase was taken and analyzed by XRPD to identify the solid phase present. Additional samplings were also carried out until only 1 solid phase remained.

[0266] Results. The results of XRPD analysis at each temperature were compared with the diffraction patterns of polymorphic forms I and II. The results indicate that polymorphic form II converts to polymorphic form I throughout the temperature range studied in n-heptane.

[0267] Cross-seeding experiment

[0268] The cross-seeding experiment was carried out in water and n-heptane.

[0269] Method. A saturated solution of polymorph II was prepared in water and n-heptane at 25 °C. Then, after 45 minutes, seeds of polymorph I were added to the suspension and stirring was continued at 25 °C. Then the solid phase in the suspension was controlled by XRPD regularly.

[0270] Result. The results of the cross-seeding experiment carried out in water and n-heptane showed that polymorph I had higher stability than polymorph II at 25 °C.

Claims

1. Crystal forms of (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid: Crystal form I or crystal form II, wherein: Crystal form I is characterized in that its X-ray powder diffraction (XRPD) pattern includes peaks at 2θ values of 13.5°, 14.0°, 14.8°, 16.0° and 18.0°; and Crystal form II is characterized in that its XRPD pattern includes peaks at 2θ values of 10.8°, 12.1°, 12.4° and 22.3°.

2. The crystal form I according to claim 1, wherein its XRPD pattern includes peaks at 2θ values of 7.2°, 12.8°, 13.5°, 14.0°, 14.5°, 14.8°, 16.0°, 16.7°, 17.4°, 18.0°, 18.9°, 19.9°, 20.4° and 23.2°.

3. The crystal form I according to claim 1 or claim 2, which has an XRPD pattern substantially as shown in Figure 1.

4. The crystal form I according to any one of claims 1 to 3, wherein its differential scanning calorimetry (DSC) thermogram shows an endothermic peak with a peak temperature of about 183 °C and a start temperature of about 180 °C.

5. The crystal form I according to any one of claims 1 to 4, wherein its unit cell parameters are equal to: Unit cell dimensions: α,β,γ = 90° Space group: P212121 Number of molecules per unit cell: 4 Unit cell volume: Density (calculated value): 1.19 g / cm 3 .

6. The crystal form II according to claim 1, wherein its XRPD pattern includes peaks at 2θ values of 10.8°, 12.1°, 12.4°, 15.3°, 16.2°, 18.5°, 19.5°, 20.7°, 21.5° and 22.3°.

7. The crystal form II according to claim 1 or claim 6, which has an XRPD pattern substantially as shown in Figure 5.

8. The crystal form II according to any one of claims 1, 6 or 7, wherein its DSC thermogram shows a first endothermic peak with a peak temperature of about 110 °C, starting at about 100 °C; an exothermic peak with a peak temperature of about 149 °C, starting at about 134 °C; and a second endothermic peak with a peak temperature of about 181 °C, starting at about 179 °C.

9. A method for preparing the crystal form I according to any one of claims 1 to 5, which comprises: 1) Dissolving (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid in an ethanol / water mixture at a temperature of at most 80 °C, wherein the ethanol / water ratio is from 100 / 0 to 5 / 95; 2) If necessary, adding water to bring the ethanol / water ratio to from 95 / 5 to 5 / 95, wherein the water is preferably heated at the same temperature as the ethanol used in step 1); and 3) Cooling the mixture, preferably to a temperature of 0 °C to 10 °C, and maintaining this temperature for a period suitable for recovering crystal form I.

10. A method for preparing the crystal form II according to claim 1 or any one of claims 6 to 8, which comprises: 1) Dissolve (2S,5R)-5-(2-chlorophenyl)-1-(2'-methoxy-[1,1'-biphenyl]-4-carbonyl)pyrrolidine-2-carboxylic acid in acetonitrile at a temperature of at most 80 °C; and 2) Cool the mixture of step 1), preferably to room temperature, and maintain this temperature for a period suitable for recovering polymorph II.

11. A pharmaceutical composition comprising the polymorph according to any one of claims 1 to 8 and at least one pharmaceutically acceptable carrier.

12. The polymorph according to any one of claims 1 to 8 for use as a medicament.

13. The polymorph according to any one of claims 1 to 8 for the treatment of inflammatory diseases, wherein the inflammatory diseases are selected from rheumatoid arthritis; inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis; colitis; collagenous colitis; lymphocytic colitis; immune-related enterocolitis; immune-mediated colitis (IMC); pouchitis; celiac disease; irritable bowel syndrome; gut microbiota dysbiosis; Paget's disease; osteoporosis; multiple myeloma; uveitis; acute myeloid leukemia, chronic myeloid leukemia; pancreatic β-cell destruction; rheumatoid spondylitis; osteoarthritis; gouty arthritis and other arthritic conditions; gout; adult respiratory distress syndrome (ARDS); chronic pulmonary inflammatory diseases; silicosis; pulmonary sarcoidosis; psoriasis; rhinitis; allergic reactions; contact dermatitis; pancreatitis; allergy; hepatitis; asthma; muscle degeneration; cachexia; Reiter's syndrome; type I diabetes; bone resorption diseases; graft-versus-host reaction; ischemia-reperfusion injury; traumatic brain injury; multiple sclerosis; autoimmune brain diseases, encephalitis, encephalomyelitis; cerebral malaria; sepsis; septic shock; toxic shock syndrome; endotoxin shock; Gram-negative sepsis; fever and myalgia caused by infection; heartburn; inflammatory conditions triggered by the release of anorectic gut hormones; microbiome-related neurological and mood disorders and neurodegenerative diseases characterized by neuroinflammation.

14. The polymorph according to any one of claims 1 to 8 for the treatment of inflammatory diseases, wherein the inflammatory diseases are selected from inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, colitis, collagenous colitis, lymphocytic colitis, immune-related enterocolitis including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1, immune-mediated colitis (IMC) including as an adverse event of cancer therapy in response to the use of checkpoint inhibitors such as inhibitors of CTLA-4, PD-1, PD-L1, pouchitis, celiac disease, irritable bowel syndrome, gut microbiota dysbiosis, type 1 diabetes, multiple sclerosis, autoimmune brain diseases, encephalitis and encephalomyelitis.

15. The crystalline form according to any one of claims 1 to 8, which is used for treating cancer, wherein the cancer is preferably selected from lung cancer, non-small cell lung cancer, small cell lung cancer, breast cancer, prostate cancer, ovarian cancer, endometrial cancer, vaginal cancer, testicular cancer, cervical cancer, bladder cancer, head and neck cancer, kidney cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, thyroid cancer, gastrointestinal cancer, colorectal cancer, gastric cancer, colon cancer, liver cancer, leukemia, lymphoma, skin cancer, melanoma, multiple myeloma, glioma, glioblastoma, mesothelioma, retinoblastoma, sarcoma, Ewing's sarcoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, bone cancer and gastric cardia cancer.

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