New crystalline forms of nitrogen-containing tricyclic compounds and uses thereof

Abstract

The present application belongs to the technical field of drugs, and relates to a new crystal form of a nitrogen-containing tricyclic compound and a use thereof. Specifically, the present application relates to a new crystal form of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepino[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid, which is a crystal form N. The present application also relates to a pharmaceutical composition comprising the crystal form. The crystal form or the pharmaceutical composition can be used for preventing, treating or alleviating a disease mediated by FXR in a patient.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to novel crystalline forms of nitrogen-containing tricyclic compounds and their uses. Specifically, it relates to a novel crystalline form of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxazo[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid and its pharmaceutical compositions, and further relates to the use of the novel crystalline form or the pharmaceutical composition in the preparation of medicines, particularly in the preparation of medicines for the prevention, treatment, or relief of FXR-mediated diseases in patients. The crystalline form or the pharmaceutical composition described in this invention can be used for the prevention, treatment, or relief of FXR-mediated diseases in patients. Background Technology

[0002] Patent applications WO 2018024224A1 and CN107686486A disclose nitrogen-containing tricyclic compounds that can be used as FXR activity modifiers, their preparation methods, and applications. Specifically, compound 7 is disclosed, namely, compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxazo[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid (the compound shown in formula (I)).

[0003]

[0004] Patent applications WO2021104427A1, WO2021104421A1 and CN112876490A disclose the crystal form of this compound.

[0005] It is well known in the art that drug polymorphism is a common phenomenon in drug development and an important factor affecting drug quality. Different polymorphs of the same drug may differ significantly in appearance, solubility, melting point, dissolution rate, and bioavailability, and may also have different impacts on drug stability, bioavailability, and efficacy. Therefore, a comprehensive investigation of drug polymorphism is necessary in drug development. Summary of the Invention

[0006] This invention provides a novel crystalline form of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxazo[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid (the compound shown in formula (I)). The novel crystalline form is crystalline form N, which is stable, has little or no hygroscopicity, and possesses good pharmacological properties (e.g., good pharmacokinetic properties), thus exhibiting excellent drug-like properties. This invention also provides pharmaceutical compositions comprising crystalline form N of the compound shown in formula (I).

[0007] Specifically, the present invention relates to crystalline form N of the compound represented by formula (I) and pharmaceutical compositions thereof, and the use of said crystalline form N or said pharmaceutical composition in the preparation of medicaments for the prevention, treatment or relief of FXR-mediated diseases in patients. Crystalline form N of the present invention may also be in solvate form, such as hydrate form.

[0008] On the one hand, the present invention provides a crystal form of the compound shown in formula (I), wherein the crystal form is crystal form N.

[0009]

[0010] In some embodiments, the X-ray powder diffraction pattern of the N crystal form of the present invention has diffraction peaks at the following 2θ angles: 13.05°, 15.31°, 18.32°, 23.93°, 26.21°, 27.83°, wherein the error range of each peak is ±0.2°.

[0011] In some embodiments, the X-ray powder diffraction pattern of crystal form N of the present invention has diffraction peaks at the following 2θ angles: 12.22°, 13.05°, 13.64°, 15.31°, 18.32°, 18.47°, 22.67°, 23.93°, 24.47°, 24.93°, 26.21°, 27.83°, 37.60°, wherein the error range of each peak is ±0.2°.

[0012] In other embodiments, the X-ray powder diffraction pattern of crystal form N according to the present invention also has characteristic peaks at at least one of the following 2θ angles: 14.03°, 16.15°, 19.47°, 21.24°, 22.77°, 24.58°, 25.25°, 26.88°, 29.63°, 30.34°, 30.72°, wherein the error range of each peak is ±0.2°.

[0013] In some other embodiments, the X-ray powder diffraction pattern of crystal form N of the present invention also has characteristic peaks at at least one of the following 2θ angles: 6.13°, 7.67°, 17.91°, 19.16°, 19.95°, 21.70°, 22.16°, 23.21°, 24.17°, 25.81°, 27.41°, 28.28°, 30.09°, 30.46°, 30.90°, 33.81°, 35.30°, 36.23°, 40.50°, and 45.36°, wherein the error range of each peak is ±0.2°.

[0014] In some embodiments, the X-ray powder diffraction pattern of the N crystal form of the present invention has diffraction peaks at the following 2θ angles: 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 18.32°, 18.47°, 19.47°, 21.24°, 22.67°, 22.77°, 23.93°, 24.47°, 24.58°, 24.93°, 25.25°, 26.21°, 26.88°, 27.83°, 29.63°, 30.34°, 30.72°, 37.60°, wherein the error range of each peak is ±0.2°.

[0015] In some embodiments, the X-ray powder diffraction pattern of crystal form N described in this invention has diffraction peaks at the following 2θ angles: 6.13°, 7.67°, 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 17.91°, 18.32°, 18.47°, 19.16°, 19.47°, 19.95°, 21.24°, 21.70°, 22.16°, 22.67°, 22.77°, 23.21°. The peaks are 23.93°, 24.47°, 24.58°, 24.93°, 25.25°, 25.81°, 26.21°, 26.88°, 27.41°, 27.83°, 28.28°, 29.63°, 30.09°, 30.34°, 30.46°, 30.72°, 30.90°, 33.81°, 35.30°, 36.23°, 37.60°, 40.50°, and 45.36°, with an error range of ±0.2° for each peak.

[0016] In some embodiments, the X-ray powder diffraction pattern of crystal form N according to the present invention has diffraction peaks at the following 2θ angles: 6.13°, 7.67°, 9.23°, 11.08°, 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 17.91°, 18.32°, 18.47°, 19.16°, 19.47°, 19.95°, 20.26°, 21.24°, 21.70°, 22.16°, 22.67°, 22.77°, 23.0°. The peaks are 2°, 23.21°, 23.93°, 24.47°, 24.58°, 24.93°, 25.25°, 25.81°, 26.21°, 26.88°, 27.41°, 27.83°, 28.28°, 29.63°, 30.09°, 30.34°, 30.46°, 30.72°, 30.90°, 33.81°, 35.30°, 36.23°, 37.60°, 40.50°, 41.14°, and 45.36°, with an error range of ±0.2° for each peak.

[0017] In some embodiments, the X-ray powder diffraction pattern of crystal form N described in this invention has diffraction peaks at the following 2θ angles: 6.13°, 7.67°, 9.23°, 11.08°, 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 17.12°, 17.91°, 18.32°, 18.47°, 19.16°, 19.47°, 19.95°, 20.26°, 20.52°, 21.24°, 21.70°, 22.16°, 22.67°, 22.77°, 23.02°, 23.21°, 23.93°, 24.17°, 24.47°, 24.58°. The peaks are 24.93°, 25.25°, 25.81°, 26.21°, 26.88°, 27.41°, 27.83°, 28.28°, 29.63°, 30.09°, 30.34°, 30.46°, 30.72°, 30.90°, 31.06°, 31.59°, 32.12°, 32.58°, 33.16°, 33.81°, 34.84°, 35.30°, 36.23°, 37.06°, 37.60°, 38.84°, 39.16°, 40.50°, 40.77°, 41.14°, 41.69°, 44.30°, and 45.36°, with an error range of ±0.2° for each peak.

[0018] In some embodiments, the X-ray powder diffraction pattern of crystal form N of the present invention has diffraction peaks at the following 2θ angles: 6.13°, 7.67°, 9.23°, 11.08°, 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 17.12°, 17.91°, 18.32°, 18.47°, 19.16°, 19.47°, 19.95°, 20°. 26°, 20.52°, 21.24°, 21.70°, 22.16°, 22.67°, 22.77°, 23.02°, 23.12°, 23.21°, 23.93°, 24.17°, 24.47°, 24.58°, 24.93°, 25.25°, 25.81°, 26.21°, 26.88°, 27.41°, 27.83°, 28.28°, 29.63°, 3 0.09°, 30.34°, 30.46°, 30.72°, 30.90°, 31.06°, 31.59°, 32.12°, 32.58°, 33.16°, 33.81°, 34.42°, 34.84°, 35.30°, 35.68°, 36.23°, 36.68°, 37.06°, 37.60°, 38.21°, 38.84°, 39.16°, 40.02° The peaks are 40.50°, 40.77°, 41.14°, 41.69°, 42.31°, 43.66°, 44.30°, 45.36°, 46.28°, 47.66°, 48.89°, 50.35°, 51.00°, 51.88°, 52.41°, 53.69°, 54.09°, 56.78°, 57.80°, and 59.49°, with an error range of ±0.2° for each peak.

[0019] In some embodiments, the crystal form N of the present invention has substantially the following characteristics: Figure 1 The X-ray powder diffraction pattern shown.

[0020] In some embodiments, the differential scanning calorimetry (DSC) of crystal form N of the present invention includes an endothermic peak at 203.58 °C ± 3 °C.

[0021] In some embodiments, the crystal form N of the present invention has substantially the following characteristics: Figure 2 The differential scanning calorimeter shown is a thermal image.

[0022] In some embodiments, the crystalline N of the present invention loses approximately 0.0336% of its weight under heating conditions, with an error tolerance of ±0.1%.

[0023] In some embodiments, the crystal form N of the present invention has substantially the following characteristics: Figure 3 The thermogravimetric analysis curves are shown below.

[0024] On the other hand, the present invention also relates to a method for preparing the crystal form N of the compound shown in formula (I), comprising: adding the compound shown in formula (I) to a solvent, heating to dissolve, cooling to crystallize, and filtering to obtain the crystal form N.

[0025] In some embodiments, the solvent in the preparation method of the present invention is an organic solvent, including but not limited to alcohol solvents such as ethanol or isopropanol, and ester solvents such as ethyl acetate.

[0026] In some embodiments, the heating in the preparation method of the present invention refers to heating to reflux. In some embodiments, the heating in the preparation method of the present invention refers to heating to a high temperature, such as heating to 65°C or above, or heating to 70°C or above.

[0027] In some embodiments, after heating to dissolve the compound shown in formula (I), the mixture is kept at that temperature for a period of time, such as 0.5 hours or 1 hour.

[0028] Optionally, seed crystals can be used in the preparation method, for example, seed crystals of crystal form N added after a period of heat preservation. In some embodiments, the temperature can be lowered before adding the seed crystals, for example, the temperature of the reaction solution can be lowered to 60°C, 45°C, or 25°C. In some embodiments, the reaction solution is cooled after a period of heat preservation, and then the seed crystals are added.

[0029] In some embodiments, in the preparation method of the present invention, cooling refers to cooling to room temperature or cooling to a low temperature (e.g., -10°C).

[0030] In some embodiments, the preparation method of the present invention further includes filtration followed by drying to obtain the target crystal form. The drying includes room temperature vacuum drying or high-temperature (e.g., 60°C) vacuum drying.

[0031] In some embodiments, the method for preparing the crystal form N of the compound represented by formula (I) of the present invention includes: dissolving the compound represented by formula (I) in an organic solvent, heating and refluxing until the solid dissolves, maintaining the temperature, cooling to room temperature, continuing to stir, filtering, and drying to obtain crystal form N; wherein the organic solvent includes isopropanol, etc.

[0032] In some embodiments, the method for preparing the crystal form N of the compound of formula (I) of the present invention includes: dissolving the compound of formula (I) in an organic solvent, heating to reflux until the solid dissolves, holding at the temperature, cooling to a slightly lower temperature, adding seed crystals, cooling to an even lower temperature, continuing to stir, filtering, and drying to obtain crystal form N; wherein the organic solvent includes ethyl acetate or ethanol, etc.; the slightly lower temperature is a temperature below the reflux temperature, such as 60°C, 45°C, or 25°C, etc.; the even lower temperature is a temperature below the slightly lower temperature, such as room temperature or a temperature below 0°C (e.g., -10°C), etc.

[0033] On the one hand, the present invention also provides a pharmaceutical composition comprising the crystal form described herein, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, or combination thereof.

[0034] On the other hand, the present invention also relates to the use of the crystal form of the compound represented by formula (I) or the pharmaceutical composition in the preparation of a medicament, wherein the medicament is used to prevent, treat or alleviate a patient’s FXR-mediated disease.

[0035] On one hand, the present invention relates to methods for preventing, treating, or alleviating FXR-mediated diseases in patients, including administering medication to patients at a pharmaceutically acceptable effective dose of the crystal form or pharmaceutical composition described herein; further, the method includes administering an effective therapeutic dose of the crystal form N or pharmaceutical composition described herein to humans or animals.

[0036] On the other hand, the crystal form of the compound represented by formula (I) of the present invention or the pharmaceutical composition thereof may be used to prevent, treat or alleviate FXR-mediated diseases in patients.

[0037] In some embodiments, the FXR-mediated diseases described in this invention are cardiovascular and cerebrovascular diseases, diseases related to dyslipidemia, metabolic syndrome, proliferative disorders, fibrosis, inflammatory diseases, or diseases related to the liver and gallbladder.

[0038] In other embodiments, the cardiovascular and cerebrovascular diseases described in this invention are atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke, or thrombosis.

[0039] In other embodiments, the metabolic syndrome described in this invention includes insulin resistance, hyperglycemia, hyperinsulinemia, elevated levels of fatty acids or triglycerides in the blood, hyperlipidemia, obesity, hypertriglyceridemia, hypercholesterolemia, syndrome X, diabetic complications, atherosclerosis, hypertension, acute anemia, neutropenia, dyslipidemia, type II diabetes, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, dyslipidemia, or a combination of diabetes and abnormally high body mass index.

[0040] In other embodiments, the proliferative diseases described in this invention are hepatocellular carcinoma, colonic adenoma, polyposis, colonic adenocarcinoma, breast cancer, membranous adenocarcinoma, Bartholin's esophageal cancer, and other forms of gastrointestinal or liver neoplastic diseases.

[0041] In other embodiments, the fibrosis, inflammatory disease, or hepatobiliary-related disease described in this invention includes non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, cholestasis, liver fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial cholestasis, cystic fibrosis, drug-induced bile duct injury, gallstones, cirrhosis, hepatitis B, sebaceous gland disease, alcoholic cirrhosis, biliary obstruction, cholelithiasis, colitis, neonatal jaundice, kernicterus, or intestinal bacterial overgrowth.

[0042] On the one hand, the present invention also relates to a method for preparing the crystal form of the compound shown in formula (I).

[0043] The solvents used in the crystal form preparation method described in this invention are not particularly limited; any solvent capable of dissolving the starting material to a certain extent without affecting its properties is included in this invention. Furthermore, many similar modifications, equivalent substitutions, or solvents, solvent combinations, and different proportions of solvent combinations described in this invention are considered to be within the scope of this invention. This invention provides preferred solvents for each reaction step.

[0044] The preparation experiments of the crystal form described in this invention will be described in detail in the Examples section. Simultaneously, this invention provides property testing experiments for the crystal form N, such as pharmacokinetic experiments, stability experiments, and hygroscopicity experiments. The experimental results show that the crystal form N of the compound represented by formula (I) of this invention has good biological activity and high stability, making it suitable for pharmaceutical applications. Specifically, the crystal form N of this invention has superior pharmacokinetic properties, for example, a higher exposure level; the crystal form N of this invention is very stable under high temperature, high humidity, and / or light conditions, and its crystal structure remains essentially unchanged; furthermore, the crystal form N of this invention is not easily deliquescent due to high humidity, facilitating long-term storage of the drug.

[0045] Compared with the prior art, the crystal form N described in this invention has superior technical effects in at least one aspect, such as having better pharmacokinetic properties (e.g., higher exposure and / or higher Cmax value) and / or higher stability.

[0046] Definitions and general terms

[0047] Unless otherwise stated, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. All patents and publications related to this invention are incorporated herein by reference in their entirety. Although any methods and substances similar to or identical to those described herein may be used in the practice or testing of this invention, preferred methods, apparatus, and substances are described herein.

[0048] "Crystal form" or "crystalline shape" refers to a solid having a highly regular chemical structure, including, but not limited to, single-component or multi-component crystals, and / or polymorphs of compounds, solvates, hydrates, inclusion compounds, eutectics, salts, solvates of salts, and hydrates of salts. The crystalline form of a substance can be obtained by many methods known in the art. These methods include, but are not limited to, melt crystallization, melt cooling, solvent crystallization, crystallization in a confined space, such as in nanopores or capillaries, crystallization on a surface or template, such as on a polymer, crystallization in the presence of additives such as co-crystallized antimolecules, desolventization, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, reactive crystallization, antisolvent addition, grinding, and solvent drop grinding, etc.

[0049] "Solvent" refers to a substance (typically a liquid) that can completely or partially dissolve another substance (typically a solid). Solvents used in the implementation of this invention include, but are not limited to, water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, dichloromethane, dimethyl sulfoxide, 1,4-dioxane, ethanol, ethyl acetate, butanol, tert-butanol, N,N-dimethylacetamide, N,N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, mixtures thereof, etc.

[0050] An antisolvent is a fluid that promotes the precipitation of a product (or product precursor) from a solvent. Antisolvents can include cold gases, fluids that promote precipitation through chemical reactions, or fluids that reduce the solubility of a product in a solvent; they can be the same liquid as the solvent but at a different temperature, or they can be a different liquid from the solvent.

[0051] A "solvent" is a compound that has a solvent on its surface, in its crystal lattice, or both on its surface and in its crystal lattice. The solvent can be water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, dichloromethane, dimethyl sulfoxide, 1,4-dioxane, ethanol, ethyl acetate, butanol, tert-butanol, N,N-dimethylacetamide, N,N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, methyl pyrrolidone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, and mixtures thereof. A specific example of a solvate is a hydrate, in which water is the solvent on its surface, in its crystal lattice, or both on its surface and in its crystal lattice. A hydrate may or may not have other solvents besides water on its surface, in its crystal lattice, or both on its surface and in its crystal lattice.

[0052] Crystal forms can be identified using various techniques, such as X-ray powder diffraction (XRPD), infrared absorption spectroscopy (IR), melting point method, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), nuclear magnetic resonance, Raman spectroscopy, X-ray single crystal diffraction, calorimetry, scanning electron microscopy (SEM), quantitative analysis, solubility and dissolution rate, etc.

[0053] X-ray powder diffraction (XRPD) can detect changes in crystal form, crystallinity, and crystal structure, and is a commonly used method for identifying crystal forms. In some embodiments, the crystal form of the present invention is characterized by an XRPD pattern with certain peak positions, which is essentially as shown in the XRPD pattern provided in the accompanying drawings. However, the measurement of 2θ in the XRPD pattern can have experimental errors; the measurement of 2θ in the XRPD pattern may differ slightly between different instruments and different samples. Therefore, the value of 2θ cannot be considered absolute. Based on the instrument used in this experiment, there is an error tolerance of ±0.2° for the diffraction peaks.

[0054] Differential scanning calorimetry (DSC) is a technique that measures the energy difference between a sample and an inert reference (commonly α-Al₂O₃) as a function of temperature by continuously heating or cooling under programmed control. In some embodiments, the crystal form described in this invention is characterized by a DSC pattern with characteristic peak positions, which is essentially as shown in the DSC pattern provided in the accompanying drawings. However, DSC patterns can be subject to experimental error; the peak positions and peak values ​​may vary slightly between different instruments and different samples. Therefore, the peak positions or peak values ​​of the endothermic peaks in the DSC pattern cannot be considered absolute. Based on the instrument used in this experiment, there is an error tolerance of ±3° for the endothermic peaks.

[0055] Thermogravimetric analysis (TGA) is a technique used under programmed control to determine the change in mass of a substance with temperature. It is suitable for examining the loss of solvent in crystals or the sublimation and decomposition of samples, and can infer the presence of water of crystallization or crystallization solvent in the crystal. The mass change shown by the TGA curve depends on many factors, including sample preparation and instrumentation; the mass change detected by TGA varies slightly between different instruments and different samples. Based on the instrument used in this experiment, there is an error tolerance of ±0.1% for the mass change.

[0056] In the context of this invention, the 2θ values ​​in X-ray powder diffraction patterns are all in degrees (°).

[0057] The term “basically as shown” means that at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% of the peaks are shown in the X-ray powder diffraction pattern or DSC pattern.

[0058] When referring to a spectrum or / and the data appearing in the graph, a "peak" refers to a feature that a person skilled in the art can identify and that is not attributable to background noise.

[0059] The present invention relates to a novel crystal form of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxazo[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid (the compound shown in formula (I)), which exists in a substantially pure crystalline form.

[0060] "Substantially pure" means that a crystal form substantially contains no other crystal forms, i.e., the purity of the crystal form is at least 80%, or at least 85%, or at least 90%, or at least 93%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9%, or the crystal form contains other crystal forms whose percentage in the total volume or total weight of the crystal form is less than 20%, or less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01%.

[0061] "Substantially free of" means that one or more other crystal forms account for less than 20%, or less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01% of the total volume or weight of the crystal form.

[0062] In an XRPD pattern, "relative intensity" (or "relative peak height") refers to the ratio of the intensity of the other peaks to the intensity of the first strongest peak when the intensity of the first strongest peak is 100%.

[0063] In the context of this invention, when the terms "about" or "approximately" are used, whether or not they are used, it means within 10% of a given value or range, appropriately within 5%, and particularly within 1%. Alternatively, for those skilled in the art, the term "about" or "approximately" means within an acceptable standard error of the average. Whenever a number with a value of N is disclosed, any number having a value within N+ / -1%, N+ / -2%, N+ / -3%, N+ / -5%, N+ / -7%, N+ / -8%, or N+ / -10% is explicitly disclosed, where "+ / -" refers to addition or subtraction.

[0064] In this invention, "room temperature" refers to a temperature from about 10°C to about 40°C. In some embodiments, "room temperature" refers to a temperature from about 20°C to about 30°C; in other embodiments, "room temperature" refers to 20°C, 22.5°C, 25°C, 27.5°C, etc.

[0065] Pharmaceutical compositions, formulations, administration and uses of the crystal form described in this invention

[0066] The pharmaceutical compositions of the present invention are characterized by a crystal form of the compound represented by formula (I) or any combination thereof and a pharmaceutically acceptable carrier, excipient, or excipient. The amount of the crystal form of the compound in the pharmaceutical compositions of the present invention can effectively and detectably treat or alleviate FXR-mediated diseases in patients.

[0067] As described in this invention, pharmaceutically acceptable compositions of this invention further comprise pharmaceutically acceptable carriers, excipients, or excipients, such as those used in this invention, including any solvent, diluent, or other liquid excipient, dispersant or suspending agent, surfactant, isotonic agent, thickener, emulsifier, preservative, solid binder or lubricant, etc., suitable for a particular target dosage form. As described in the following literature: In Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. D.B. Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J.C. Boylan, 1988-1999, Marcel Dekker, New York, the contents of this literature demonstrate that different carriers can be used in the formulation of pharmaceutically acceptable compositions and their known methods of preparation. The use of any conventional carrier medium that is incompatible with the compounds or their crystal forms of the present invention, such as any adverse biological effects or harmful interactions with any other component of a pharmaceutically acceptable composition, is also within the scope of this invention.

[0068] The crystal form described in this invention can be uniformly combined in a mixture as an active ingredient with a drug carrier according to conventional drug compounding techniques. Depending on the required dosage form, such as oral or parenteral (including intravenous) administration, the carrier can be of various forms. When preparing compositions for oral dosage forms, any conventional pharmaceutical medium can be used, for example, water, ethylene glycol, oil, alcohol, flavoring agents, preservatives, coloring agents, etc., used in the preparation of oral liquid dosage forms such as suspensions, elixirs, and solutions; or starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc., used in the preparation of oral solid dosage forms such as powders, hard capsules, soft capsules, and tablets, wherein solid oral dosage forms are preferred over liquid dosage forms.

[0069] Because tablets and capsules are easy to take, they represent the most advantageous form of oral dosage unit, and solid drug carriers are obviously used in this case. If necessary, tablets can be coated with standard aqueous solutions or non-aqueous techniques.

[0070] A variety of other materials can be used as coatings or to alter the shape of the dosage units. For example, tablets can be coated with shellac, sugar, or both. In addition to the active ingredient, syrups or elixirs may contain sucrose as a sweetener, methylparaben or propylparaben as preservatives, dyes, and flavorings (e.g., cherry or orange flavor).

[0071] The crystal forms described in this invention can also be administered parenterally. Solutions or suspensions of these active substances can be prepared by appropriate mixing with surfactants (such as hydroxypropyl cellulose) in water. Dispersants can also be prepared in glycerol, liquid polyethylene glycol and mixtures thereof, and in oils. Under normal storage and use conditions, these formulations contain preservatives to prevent microbial growth.

[0072] Suitable pharmaceutical forms for injection include sterile aqueous solutions or dispersants and sterile powders for the immediate preparation of sterile injectable solutions or dispersants. In all cases, the pharmaceutical form must be sterile and must be a fluid in an easily injectable form. It must be stable under the conditions of manufacture and storage and must be preserved under conditions resistant to contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

[0073] The crystal form described in this invention can be delivered to mammals, especially humans, using any suitable method of administration. For example, it can be administered orally, rectally, topically, parenterally, ocularly, pulmonaryly, or nasally. Dosage forms include tablets, lozenges, dispersants, suspensions, solutions, capsules, emulsions, ointments, aerosols, etc.

[0074] The crystal form or pharmaceutical composition thereof disclosed in this invention is effective in preventing, treating, or alleviating FXR-mediated diseases in patients, particularly in treating non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), obesity, hypertriglyceridemia, atherosclerosis, chronic intrahepatic cholestasis, primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), progressive familial cholestasis (PFIC), drug-induced bile duct injury, gallstones, cirrhosis, hepatitis B, and sebaceous gland disease. Alcohol-induced cirrhosis, cystic fibrosis, biliary obstruction, gallstones, liver fibrosis, dyslipidemia, atherosclerosis, type II diabetes, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, peripheral arterial occlusive disease (PAOD), colitis, neonatal jaundice, kernicterus, venous occlusive disease, portal hypertension, metabolic syndrome, acute myocardial infarction, acute stroke, thrombosis, hypercholesterolemia, intestinal bacterial overgrowth, erectile dysfunction, gastrointestinal neoplasia, and liver neoplasia, etc. Attached Figure Description

[0075] Figure 1 This is an X-ray powder diffraction (XRPD) pattern of the N crystal form described in this invention.

[0076] Figure 2 This is a differential scanning calorimetry (DSC) image of the N-type crystal described in this invention.

[0077] Figure 3 This is the thermogravimetric analysis (TGA) diagram of the N crystal form described in this invention.

[0078] Figure 4 This is a dynamic water adsorption (DVS) diagram of the N crystal form described in this invention. Detailed Implementation

[0079] The present invention will be further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein.

[0080] The X-ray powder diffraction analysis method used in this invention is as follows: An Empyrean diffractometer is used, employing Cu-Kα radiation (45 kV, 40 mA) to obtain the X-ray powder diffraction pattern. The powdered sample is prepared into a thin layer on a single-crystal silicon sample holder, placed on a rotating sample stage, and analyzed in 0.0167° steps within the range of 3°–60°. Data is collected using Data Collector software, processed using High Score Plus software, and read using Data Viewer software.

[0081] The differential scanning calorimetry (DSC) analysis method used in this invention is as follows: Differential scanning calorimetry is performed using a TA Q2000 module with a thermal analysis controller. Data is collected and analyzed using TA Instruments Thermal Solutions software. Approximately 1-5 mg of sample is accurately weighed into a specially designed aluminum crucible with a lid, and sample analysis is performed using a linear heating device at 10°C / min, from room temperature to approximately 300°C. During use, the DSC chamber is purged with dry nitrogen.

[0082] The thermogravimetric analysis (TGA) method used in this invention is as follows: TGA analysis is performed using a TA Q500 module equipped with a thermal analysis controller. Data is collected and analyzed using TA Instruments Thermal Solutions software. Approximately 10-30 mg of sample is placed in a platinum crucible and analyzed from room temperature to approximately 300°C using a linear heating device at 10°C / min. During use, the DSC chamber is purged with dry nitrogen.

[0083] The hygroscopicity of this invention was measured using a DVS INT-Std dynamic moisture and gas adsorption analyzer from Surface Measurement Systems, UK. The humidity test range was 0%-95%, the airflow was 200 mL / min, the temperature was 25°C, and the test point was one point for every 5% humidity.

[0084] Specific implementation methods

[0085] Original sample: The compound (2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxazo[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid) of formula (I) was prepared by the method of Example 9 in patent application CN107686486A.

[0086] Example

[0087] Example 1 Crystal form of compound (I) N

[0088] 1. Preparation of crystalline N

[0089] Method 1: The original sample (0.21 g) and 9-fluorenone (0.07 g) were suspended in isopropanol (2.0 mL), heated to reflux until the solid was completely dissolved, kept warm for 1 hour, cooled naturally to room temperature, stirred overnight, filtered, and the filter cake was vacuum dried at room temperature for 6 hours to obtain an off-white solid (0.12 g, yield 57.1%).

[0090] Method 2: The original sample (5.0 g) and 9-fluorenone (1.8 g) were suspended in ethyl acetate (75.0 mL), heated to 70 °C until the solid was completely dissolved, kept at this temperature for 1 hour, cooled to 25 °C, and seed crystals of crystal form N (0.1 g) were added. After the addition was complete, the mixture was cooled to -10 °C and stirred overnight. The mixture was then filtered, and the filter cake was washed with n-heptane (30.0 mL) and dried under vacuum at room temperature for 4 hours to obtain a white solid (2.34 g, yield 46.8%).

[0091] Method 3: The original sample (1.05 g) and 9-fluorenone (0.36 g) were suspended in ethanol (20.0 mL), heated to reflux until the solid was completely dissolved, kept at this temperature for 0.5 hours, cooled to 45 °C, and seed crystals of crystalline form N (0.02 g) were added. The mixture was then allowed to cool naturally to room temperature and stirred for 4 hours. The mixture was then filtered, and the filter cake was washed with n-heptane (10.0 mL). The mixture was then dried under vacuum at 60 °C for 4 hours to obtain a white solid (0.77 g, yield 73.3%).

[0092] Method 4: The original sample (1.0 g) was suspended in ethanol (20.0 mL), heated to reflux, and kept warm for 0.5 hours after the solid was dissolved. Then, the temperature was lowered to 60 °C, and seed crystals of crystal form N (0.02 g) were added. The mixture was then allowed to cool naturally to room temperature and stirred for 4 hours to obtain a white solid (0.76 g, yield 76.0%).

[0093] 2. Identification of N crystal form

[0094] (1) Identification by Empyrean X-ray powder diffraction (XRPD): Using Cu-Kα radiation, the following characteristic peaks, expressed in terms of angle 2θ, were observed: 6.13°, 7.67°, 9.23°, 11.08°, 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 17.12°, 17.91°, 18.32°, 18.47°, 19.16°, 19.47°, 19.95°, 2 0.26°, 20.52°, 21.24°, 21.70°, 22.16°, 22.67°, 22.77°, 23.02°, 23.12°, 23.21°, 23.32°, 23.48°, 23.93°, 24.17°, 24.47°, 24.58°, 24.93°, 25.25°, 25.81°, 26.21°, 26.88°, 27.41°, 27.83°, 28°. 28°, 29.63°, 30.09°, 30.34°, 30.46°, 30.72°, 30.90°, 31.06°, 31.59°, 32.12°, 32.58°, 33.16°, 33.81°, 34.42°, 34.84°, 35.30°, 35.68°, 36.23°, 36.68°, 37.06°, 37.60°, 38.21°, 38.84°, 39.16° °, 40.02°, 40.50°, 40.77°, 41.14°, 41.69°, 42.31°, 43.66°, 44.30°, 45.36°, 46.28°, 47.66°, 48.89°, 50.35°, 51.00°, 51.88°, 52.41°, 53.69°, 54.09°, 56.78°, 57.80° and 59.49°, with an error tolerance of ±0.2°.

[0095] The X-ray powder diffraction pattern of crystalline N obtained by the method of this invention is basically as follows: Figure 1 As shown.

[0096] (2) The differential scanning calorimetry (DSC) analysis by TA Q2000 confirmed that the scanning speed was 10℃ / min, and it contained an endothermic peak of 203.58℃ with an error tolerance of ±3℃.

[0097] The differential scanning calorimetry (DSC) of crystalline N obtained by the method of this invention is basically as follows: Figure 2 As shown.

[0098] (3) Thermogravimetric analysis (TGA) was performed using TA Q500: the heating rate was 10℃ / min, the weight loss was 0.0336%, and there was an error tolerance of ±0.1%.

[0099] The thermogravimetric analysis (TGA) diagram of the crystalline N obtained by the method of this invention is basically as follows: Figure 3 As shown.

[0100] Example 2: Pharmacokinetic Experiment of Crystal Form N of the Present Invention

[0101] The crystalline form N of the compound of formula (I) of the present invention is filled into capsules for oral administration.

[0102] Three male Beagle dogs weighing 8-12 kg were orally administered capsules containing the test sample at a dose of 5 mg / kg. Blood samples were collected at time points of 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 24 hours. A standard curve was established based on the sample concentration. The concentration of the test sample in the plasma samples was determined using an AB SCIEX API4000 LC-MS / MS in MRM mode, and quantitative analysis was performed. Pharmacokinetic parameters were calculated using the non-compartmental model method in WinNonLin 6.3 software based on the drug concentration-time curve. The experimental results are shown in Table 1.

[0103] Table 1. Pharmacokinetic experimental data of crystal form N described in this invention.

[0104] Test sample <![CDATA[AUC last (h*ng / ml)]]> <![CDATA[C max (ng / ml)]]> <![CDATA[T max (h)]]> Crystal form N 907 348 1.20

[0105] Experimental conclusion:

[0106] As shown in Table 1, the crystalline N described in this invention has a high exposure level in Beagle dogs and exhibits good pharmacokinetic properties.

[0107] Example 3: Stability experiment of the N crystal form described in this invention

[0108] (1) High temperature experiment Take an appropriate amount of a batch of test samples and place them in a flat weighing bottle. Spread the sample into a thin layer of ≤5mm thickness and place it at 60℃ for 30 days. Take samples on the 5th, 13th and 30th days, observe the color change of the samples, and detect the purity of the samples by HPLC.

[0109] (2) High humidity test Take an appropriate amount of a batch of test samples and place them in a flat weighing bottle. Spread the sample into a thin layer of ≤5mm thickness. Place the sample at 25℃ and RH 90%±5% for 30 days. Take samples on the 5th, 13th and 30th days, observe the color change of the samples, and detect the purity of the samples by HPLC.

[0110] (3) Light Experiment Take an appropriate amount of the test sample and place it in a flat weighing bottle, spreading it into a thin layer ≤5mm thick. Place the bottle open in a light box (with UV light) at an illuminance of 4500±500lx and UV light ≥0.7w / m². 2The samples were placed under the specified conditions for 30 days, and samples were taken on the 5th, 13th and 30th days to observe the color change of the samples. The purity of the samples was detected by HPLC.

[0111] The changes in appearance and chemical purity of the N crystal form described in this invention during the stability experiment are shown in Table 2.

[0112] Table 2. Stability experiment of crystal form N described in this invention.

[0113]

[0114] Experimental conclusion:

[0115] Under high temperature, high humidity, and / or light conditions, the appearance and chemical purity of the crystalline form N described in this invention show no significant changes, exhibiting good stability and suitability for pharmaceutical applications. Furthermore, the crystal structure of the crystalline form N described in this invention is also very stable, remaining essentially unchanged under high temperature, high humidity, and / or light conditions.

[0116] Example 4: Hygroscopicity experiment of the N crystal form described in this invention

[0117] Hygroscopicity test method one:

[0118] (1) Take a dry stoppered glass weighing bottle (outer diameter 50mm, height 15mm) and place it in a suitable constant temperature desiccator at 25℃±1℃ the day before (with ammonium chloride or ammonium sulfate saturated solution at the bottom, relative humidity 80%±2%) and weigh it accurately (m1).

[0119] (2) Take an appropriate amount of the test sample, weigh it accurately (m2), and spread it evenly in the weighing bottle. The thickness of the test sample is generally about 1 mm.

[0120] (3) Leave the weighing bottle open and place it together with the bottle cap under the above constant temperature and humidity conditions for 24 hours.

[0121] (4) After closing the weighing bottle, weigh it accurately (m3) after 24 hours and calculate: weight gain percentage % = (m3-m2-m1) / m2×100%.

[0122] The experimental results are shown in Table 3 below:

[0123] Table 3. Hygroscopicity data of crystal form N described in this invention.

[0124] <![CDATA[m1(mg)]]> <![CDATA[m2(mg)]]> <![CDATA[m3(mg)]]> Weight gain due to moisture absorption (%) 30123.05 1016.77 31138.87 0.005％

[0125] Hygroscopicity test method two:

[0126] Take an appropriate amount of the test sample and test its hygroscopicity using a dynamic moisture adsorption analyzer. The experimental results are as follows: Figure 4 As shown in the table below.

[0127] Table 4. DVS experimental data of crystal form N described in this invention.

[0128] Test sample Weight gain at 80% relative humidity / % Crystal form N 0.012％

[0129] The description of hygroscopic characteristics and the definition of hygroscopic weight gain (Chinese Pharmacopoeia 2020 Edition Appendix 9103 Guidelines for Hygroscopicity Testing of Drugs, Experimental conditions: 25℃±1℃, 80%±2% relative humidity) are as follows:

[0130] Description of hygroscopic characteristics and definition of hygroscopic weight gain

[0131]

[0132] Hygroscopicity test conclusion: Based on the hygroscopicity test results conducted using the two methods described above, the weight gain of crystalline form N described in this invention after equilibrium is less than 0.2% under a relative humidity of 80%. Referring to the definition standard for hygroscopic weight gain, this indicates no or almost no hygroscopicity. Therefore, crystalline form N described in this invention is not easily deliquescent due to high humidity, facilitating long-term storage of the drug.

[0133] The above description is merely a basic explanation of the concept of this invention, and any equivalent modifications made based on the technical solution of this invention shall fall within the protection scope of this invention.

[0134] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0135] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. The crystal form of the compound shown in formula (I), wherein the crystal form is crystal form N. (I)， in, The X-ray powder diffraction pattern of the N crystal form exhibits diffraction peaks at the following 2θ angles: 12.22°, 13.05°, 13.64°, 15.31°, 18.32°, 18.47°, 22.67°, 23.93°, 24.47°, 24.93°, 26.21°, 27.83°, and 37.60°, with an error range of ±0.2 for each peak. o .

2. The crystal form according to claim 1, wherein, The X-ray powder diffraction pattern of the N crystal form shows diffraction peaks at the following 2θ angles: 6.13°, 7.67°, 9.23°, 11.08°, 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 17.91°, 18.32°, 18.47°, 19.16°, 19.47°, 19.95°, 20.26°, 21.24°, 21.70°, 22.16°, 22.67°, 22.77°, 23.02°, 23.21°, 23.93°, 24.47°, 24.58°, 24.93°. The peaks are 25.25°, 25.81°, 26.21°, 26.88°, 27.41°, 27.83°, 28.28°, 29.63°, 30.09°, 30.34°, 30.46°, 30.72°, 30.90°, 33.81°, 35.30°, 36.23°, 37.60°, 40.50°, 41.14°, and 45.36°, with an error range of ±0.2 for each peak. o .

3. The crystal form according to claim 1, wherein, The X-ray powder diffraction pattern of the N crystal form shows diffraction peaks at the following 2θ angles: 6.13°, 7.67°, 9.23°, 11.08°, 12.22°, 13.05°, 13.64°, 14.03°, 15.31°, 16.15°, 17.12°, 17.91°, 18.32°, 18.47°, 19.16°, 19.47°, 19.95°, 20.26°, 20.52°, 21.24°, 21.70°, 22.16°, 22.67°, 22.77°, 23.02°, 23.21°, 23.93°, 24.17°. 24.47°, 24.58°, 24.93°, 25.25°, 25.81°, 26.21°, 26.88°, 27.41°, 27.83°, 28.28°, 29.63°, 30.09°, 30.34°, 30.46°, 30.72°, 30.90°, 31.06°, 31.59°, 32.12°, 32.58°, 33.16°, 33.81°, 34.84°, 35.30°, 36.23°, 37.06°, 37.60°, 38.84°, 39.16°, 40.50°, 40.77° The peaks are 41.14°, 41.69°, 44.30°, and 45.36°, with an error range of ±0.2 for each peak. o .

4. The crystal form according to claim 1, wherein, The crystal form N has an X-ray powder diffraction pattern that is essentially as shown in Figure 1.

5. The crystal form according to claim 1, wherein, The differential scanning calorimetry (DSC) image of crystal form N contains 203.58 o The endothermic peak is at C ±3 °C.

6. The crystal form according to claim 1, wherein, The crystal form N has a differential scanning calorimetry (DSC) image that is essentially as shown in Figure 2.

7. A pharmaceutical composition comprising the crystal form according to any one of claims 1-6, and a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof.

8. Use of the crystal form according to any one of claims 1-6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for the prevention, treatment or relief of a patient’s FXR-mediated disease.

9. The use according to claim 8, wherein, The diseases mediated by FXR are cardiovascular and cerebrovascular diseases, metabolic syndrome, proliferative disorders, fibrotic or inflammatory diseases.

10. The use according to claim 9, wherein, The cardiovascular and cerebrovascular diseases mentioned are atherosclerosis, acute myocardial infarction, venous occlusive disease, heart failure, peripheral arterial occlusion, stroke, or thrombosis. The metabolic syndrome is defined as insulin resistance, hyperglycemia, hyperlipidemia, obesity, diabetic complications, hypertension, or type II diabetes. The hyperproliferative diseases mentioned are hepatocellular carcinoma, colonic adenoma, polyposis, breast cancer, membranous adenocarcinoma, Bartholin's esophageal cancer, and other forms of gastrointestinal or liver neoplasms. The fibrotic or inflammatory disease is cystic fibrosis or colitis.

11. The use according to claim 10, wherein, The hypertension referred to is portal hypertension or pulmonary hypertension.

12. The use according to claim 8, wherein, The FXR-mediated diseases mentioned are hypertriglyceridemia or hypercholesterolemia.

13. The use according to claim 10, wherein, The diabetic complications mentioned are diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, or comorbidities of diabetes or abnormally high body mass index.

14. The use according to claim 8, wherein, The diseases mediated by FXR mentioned are liver and gallbladder-related diseases.

15. The use according to claim 14, wherein, The liver and gallbladder-related diseases mentioned include non-alcoholic fatty liver disease, cholestasis, liver fibrosis, primary sclerosing cholangitis, drug-induced bile duct injury, cirrhosis, hepatitis B, biliary obstruction, or gallstones.

16. The use according to claim 15, wherein, The cirrhosis mentioned refers to primary biliary cirrhosis or alcohol-induced cirrhosis.

17. The use according to claim 14, wherein, The liver and gallbladder-related disease mentioned is non-alcoholic steatohepatitis.

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