Amorphous tricyclic nitrogen-containing compounds and their uses

Abstract

The present invention relates to amorphous forms of tricyclic nitrogen-containing compounds, pharmaceutical compositions comprising the amorphous forms, and the use of the amorphous forms or the pharmaceutical compositions in the manufacture of a medicament for preventing, treating, or alleviating a disease mediated by FXR in a patient.

Description

[Technical Field]

[0001] This application claims priority from Chinese Patent Application No. 201911201723.7, filed November 29, 2019.

[0002] The present invention belongs to the technical field of pharmaceuticals and relates to an amorphous form of a tricyclic nitrogen-containing compound and uses thereof, specifically to an amorphous form of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepin[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid (compound represented by formula (I)) and uses thereof, and further to a pharmaceutical composition containing the amorphous form. The amorphous form or the pharmaceutical composition is used to prevent, treat, or alleviate a disease mediated by FXR in a patient. [Background technology]

[0003] The farnesoid X receptor (FXR) is a member of the nuclear hormone receptor superfamily and is expressed primarily in the liver, kidney, and intestine (Seol et al., Mol. Endocrinol (1995), 9:72-85; Forman et al., Cell (1995), 81:687-693). It functions as a heterodimer with the retinoid X receptor (RXR) and regulates gene transcription by binding to response elements in target gene promoters. The FXR-RXR heterodimer binds with highest affinity to the inverted repeat-1 (IR-1) response element, where hexamers that bind to the consensus receptor are separated by one nucleotide. FXR is activated by bile acids (end products of cholesterol metabolism) (Makishima et al., Science (1999), 284:1362-1365; Parks et al., Science (1999), 284:1365-1368; Wang et al., Mol. Cell. (1999), 3:543-553), and bile acids are used to inhibit cholesterol catabolism (Urizar et al., (2000) J. Biol. Chem. 275:39313-393170).

[0004] FXR is a key regulator of cholesterol homeostasis, triglyceride synthesis and adipogenesis (Crawley, Expert Opinion Ther. Patents (2010), 20:1047-1057). In addition to being a target for treating dyslipidemia, obesity, vitamin D-related diseases, enteropathy, adverse drug reactions, and hepatitis (Crawley, Expert Opinion Ther. Patents (2010), 20:1047-1057), FXR can also be a therapeutic target for hepatobiliary diseases, chronic hepatitis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), cholestasis, liver fibrosis, cirrhosis, hepatitis B, metabolic diseases, lipid metabolism disorders, carbohydrate metabolism disorders, cardiometabolic diseases, atherosclerosis, type II diabetes, and diabetic complications (Frank G. Schaap et al., Journal of Medicinal Chemistry (2005), 48:5383-5402).

[0005] Patent applications WO2018024224 and CN107686486 disclose tricyclic nitrogen-containing compounds that can be used as FXR activity modulators, as well as their preparation and application. Specifically, compound 7, i.e., 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepin[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid (compound represented by formula (I)), is disclosed. However, these patent applications do not disclose the specific microstructure of the compound. [ka]

[0006] As is well known in the art, drug polymorphism is a common phenomenon in drug research and is an important factor affecting drug quality. The same drug may have different crystalline forms with significant differences in appearance, solubility, melting point, dissolution rate, biological efficacy, etc., which may have different effects on aspects such as drug stability, bioavailability, and therapeutic effect. Therefore, the issue of drug polymorphism should be comprehensively considered in drug development.

[0007] Amorphous is one form of substance crystalline polymorphism, and is an amorphous state.The various physicochemical properties and clinical pharmacological characteristics of amorphous drugs are always different from those of common crystalline drugs.As is known in the art, the bioavailability of drugs with low water solubility can be improved by converting them into salts or amorphous forms.However, the amorphous forms of many substances are unstable, and few drugs are available in the amorphous state, so research into how to make drugs available in a stable amorphous state is equally important. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Publication No. 2018024224 [Patent Document 2] China Publication Patent No. 107686486 [Non-patent literature]

[0009] [Non-Patent Document 1] Seol et al., Mol. Endocrinol (1995), 9:72-85 [Non-patent document 2] Makishima et al., Science (1999), 284:1362-1365 [Non-patent document 3] Parks et al., Science (1999), 284: 1365-1368 [Non-patent document 4] Wang et al., MoI. Cell. (1999), 3:543-553 [Non-Patent Document 5] Urizar et al. (2000) J.Biol.Chem.275:39313-393170 [Non-patent document 6] Crawley, Expert Opinion Ther. Patents (2010), 20:1047-1057 [Non-Patent Document 7] Frank G. Schaap et al., Journal of Medicinal Chemistry (2005), 48:5383-5402 Summary of the Invention [Problem to be solved by the invention]

[0010] The present invention provides an amorphous substance of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepin[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid and a pharmaceutical composition thereof. The amorphous substance or pharmaceutical composition has good pharmacological properties (e.g., good pharmacokinetic properties) and also has significantly improved properties such as stability, making it highly suitable for pharmaceutical use. [Means for solving the problem]

[0011] Specifically, the present invention relates to an amorphous form of the compound of formula (I) and pharmaceutical compositions thereof, as well as the use of the amorphous form or pharmaceutical composition in the manufacture of a medicament for preventing, treating, or alleviating a disease mediated by FXR in a patient. The amorphous form of the present invention may be in the form of a solvate, such as a hydrate.

[0012] In one aspect, the present invention provides an amorphous form of a compound of formula (I). [ka]

[0013] In some embodiments, the amorphous material has an X-ray powder diffraction pattern substantially as shown in FIG.

[0014] In some embodiments, the amorphous material has a glass transition temperature of 92.26°C ± 3°C.

[0015] In some embodiments, the amorphous material has a differential scanning calorimetry pattern substantially as shown in FIG.

[0016] In some embodiments, when the amorphous material is heated to 150°C, its thermogravimetric analysis curve comprises a weight loss of 0.409%.

[0017] In some embodiments, the amorphous material has a thermogravimetric analysis curve substantially as shown in FIG.

[0018] In one aspect, the present invention further provides a pharmaceutical composition comprising an amorphous material according to the present invention and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, or combination thereof.

[0019] In another aspect, the present invention further relates to the use of an amorphous form of the compound of formula (I) or said pharmaceutical composition in the manufacture of a medicament for preventing, treating or ameliorating a disease mediated by FXR in a patient, said use comprising administering to a human or animal an effective therapeutic dose of the amorphous form or said pharmaceutical composition according to the present invention.

[0020] In some embodiments, the FXR-mediated disease according to the present invention is a cardio-cerebrovascular disease, a dyslipidemia-related disease, metabolic syndrome, a hyperproliferative disease, a fibrosis, an inflammatory disease, or a disease associated with the liver and gallbladder.

[0021] In some other embodiments, the cardio-cerebrovascular disease according to the present invention is atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke, or thrombosis.

[0022] In some other embodiments, the metabolic syndrome according to the present invention is insulin resistance, hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, 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 complication of diabetes and an abnormally high body mass index.

[0023] In some other embodiments, the hyperproliferative disease of the present invention is hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma, breast cancer, membranous adenocarcinoma, Barrett's esophagus cancer, and other forms of gastrointestinal or hepatic neoplastic disease.

[0024] In some other embodiments, the fibrosis, inflammatory disease, or liver and gallbladder related disease according to the present invention is non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, cholestasis, hepatic fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial cholestasis, cystic fibrosis, drug-induced bile duct damage, gallstones, cirrhosis, hepatitis B, glandular skin disease, alcoholic cirrhosis, biliary atresia, cholelithiasis, colitis, neonatal jaundice, kernicterus, or intestinal bacterial overgrowth.

[0025] In one aspect, the present invention relates to a method for preventing, treating, or ameliorating a disease mediated by FXR in a patient, comprising administering to the patient an amorphous substance according to the present invention or said pharmaceutical composition in a pharmaceutically acceptable effective amount.

[0026] In another aspect, the present invention relates to the use of an amorphous form of a compound of formula (I) or a pharmaceutical composition thereof to prevent, treat or alleviate a disease mediated by FXR in a patient.

[0027] In another aspect, the present invention further relates to a method for preparing an amorphous form of the compound of formula (I).

[0028] The solvent used in the method for producing the amorphous material according to the present invention is not particularly limited, and any solvent that dissolves the starting materials to some extent and does not affect their properties is included in the present invention. Furthermore, many similar modifications, equivalent substitutions, or solvents equivalent to those described in the present invention, solvent combinations, and various ratios of solvents are all considered to be within the scope of the present invention. The present invention provides preferred solvents for use in each reaction step.

[0029] The preparation test of the amorphous substance of the present invention is described in detail in the Examples section. At the same time, the present invention provides activity measurement tests for the amorphous substance, such as pharmacokinetic tests, stability tests, and hygroscopicity tests. As can be seen from the test results, the amorphous substance of the compound represented by formula (I) of the present invention has high biological activity and high stability, making it suitable for pharmaceutical use. Specifically, the amorphous substance of the present invention has better pharmacokinetic properties, for example, higher exposure.

[0030] The description of hygroscopic characteristics and the definition of hygroscopic mass increase (Chinese Pharmacopoeia 2015 Edition, Appendix 9103, Guidelines for Drug Hygroscopicity Testing, Testing conditions: 25°C ± 1°C, relative humidity 80% ± 2%) are as explained in the table below.

[0031] Description of hygroscopic characteristics and definition of hygroscopic weight gain [Table 1]

[0032] The amorphous substance according to the present invention is resistant to deliquescence under the influence of high humidity and can be easily stored for a long period of time as a drug.

[0033] Definitions and General Terms Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and published publications relating to this invention are incorporated herein by reference in their entirety. Although any methods and materials similar or equivalent to those described herein can be used in the practice or measurement of this invention, those described herein are the preferred methods, devices, and materials.

[0034] "Amorphous" or "amorphous form" refers to a substance formed by particles (molecules, atoms, ions) arranged non-periodically in three-dimensional space, characterized by a scattered X-ray powder diffraction pattern lacking sharp peaks. Amorphous is a special physical form of a solid substance, whose locally ordered structural features mean that it is closely related to crystalline substances. Amorphous forms of substances can be obtained by many methods known in the art. Such methods include, but are not limited to, quenching, antisolvent aggregation, ball milling, spray drying, freeze drying, wet granulation, and solid dispersion techniques.

[0035] "Solvent" refers to a substance (typically a liquid) that can completely or partially dissolve another substance (typically a solid). Solvents that may be used in the practice of the present 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, t-butyl alcohol, 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, and the like.

[0036] "Anti-solvent" means a fluid that promotes precipitation of a product (or a precursor of the product) from a solvent. An anti-solvent may include a cryogenic gas, or a fluid that promotes precipitation by chemical reaction, or a fluid that reduces the solubility of the product in the solvent. It may be the same liquid as the solvent but at a different temperature, or it may be a different liquid than the solvent.

[0037] The term "solvate" refers to a compound having a solvent on the surface, in the lattice, or on the surface and in the lattice. The solvent may be water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, dichloromethane, dimethyl sulfoxide, 1,4-dioxane, ethanol, ethyl acetate, butanol, t-butyl alcohol, N,N-dimethylacetamide, N,N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, methylpyrrolidone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, and mixtures thereof. One specific example of a solvate is a hydrate, in which the solvent on the surface, in the lattice, or on the surface and in the lattice is water. The hydrate may or may not have a solvent other than water on the surface, in the lattice, or on the surface and in the lattice of the substance.

[0038] Amorphous materials can be identified by a variety of techniques, such as, for example, X-ray powder diffraction (XRPD), infrared spectroscopy (IR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), nuclear magnetic resonance, Raman spectroscopy, and scanning electron microscopy (SEM).

[0039] Differential scanning calorimetry (DSC) is a technique for measuring the change with temperature of the energy difference between a sample and an inert reference (usually α-Al2O3) by continuously heating or cooling the sample under controlled conditions. In some embodiments, the amorphous material of the present invention is characterized by having a DSC pattern of the glassy transition substantially as shown in the DSC patterns provided in the drawings of the present invention. Furthermore, because DSC patterns have experimental errors, and the peak positions and values ​​of DSC patterns from different instruments and different samples may vary slightly, the peak positions or values ​​of the DSC endothermic peaks described above cannot be considered absolute. Depending on the conditions of the equipment used in this test, the tolerance for the endothermic peak is ±3°C.

[0040] The glassy transition refers to the transition between a highly elastic state and a glassy state of an amorphous material, and is an inherent property of the material. The corresponding transition temperature is the glass transition temperature (Tg), which is one of the important physical properties of an amorphous material. Because the glass transition is a phenomenon related to molecular motion, the glass transition temperature (Tg) depends primarily on the structure of the material and is relatively insensitive to test details. In some embodiments, the glass transition temperature (Tg) of the amorphous material of the present invention is measured by differential scanning calorimetry (DSC), and is characterized by a glass transition temperature of 92.26°C. Depending on the conditions of the equipment used in the test of the present invention, the glass transition temperature has a tolerance of ±3°C.

[0041] Thermogravimetric analysis (TGA) is a technique for measuring the change in mass of a substance with temperature under controlled procedural conditions. It is suitable for detecting the process of solvent loss in crystallization or the sublimation and decomposition of a sample, and can infer the presence of water of crystallization or crystallization solvent in the crystal. The mass change shown in the TGA curve depends on many factors, such as sample preparation and equipment, and the mass change detected by TGA for different instruments and different samples will be slightly different. Depending on the conditions of the equipment used in this test, the tolerance for mass change is ±0.1°C.

[0042] The term "substantially as shown in the figure" means that at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of the peaks in an X-ray powder diffraction pattern or a DSC pattern are represented in the figure.

[0043] As used herein, when referring to data shown in spectra and / or graphs, the term "peak" refers to a feature that one of skill in the art would recognize as not being attributable to background noise.

[0044] In the context of the present invention, whether or not phrases such as "about," "approximately," or the like are used, it means within 10%, preferably within 5%, and particularly within 1% of a given value or range. Alternatively, to those skilled in the art, the terms "about" or "approximately" mean within an acceptable standard error of the mean. When a number having a value 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 expressly disclosed, where "+ / -" means plus or minus.

[0045] In the present invention, "room temperature" refers to a temperature of about 10° C. to about 40° C. In some embodiments, "room temperature" refers to a temperature of about 20° C. to about 30° C., and in other embodiments, "room temperature" refers to a temperature of 20° C., 22.5° C., or 30° C. o C, 25 o C, 27.5 oThis means C etc.

[0046] Amorphous Pharmaceutical Compositions, Formulations, Administration, and Uses of the Invention The pharmaceutical compositions of the present invention comprise an amorphous form of the compound of formula (I) and a pharmaceutically acceptable carrier, adjuvant, or excipient, and the amount of amorphous form of the compound in the pharmaceutical compositions of the present invention allows for effective and detectable treatment of FXR-mediated disorders in patients.

[0047] As described herein, the pharmaceutically acceptable compositions of the present invention further comprise a pharmaceutically acceptable carrier, adjuvant, or excipient, which may be any solvent, diluent, or other liquid excipient, dispersing or suspending agent, surfactant, isotonicity agent, thickener, emulsifier, preservative, solid binder, or lubricant, suitable for a particular target dosage form. The following references, in combination with the present specification, reveal various carriers that can be used to formulate pharmaceutically acceptable compositions and known methods for their preparation: 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. Except insofar as any conventional carrier medium is incompatible with the compounds of the present invention or their amorphous forms, such as by producing any undesired biological effects or by interacting in a deleterious manner with any other component of the pharmaceutically acceptable composition, such use is within the scope of the present invention.

[0048] The amorphous substance of the present invention can be used as an active ingredient in a homogeneous mixture with a drug carrier prepared by conventional drug compounding techniques. The carrier can take various forms depending on the dosage form required for administration, e.g., oral or parenteral (including intravenous). When preparing a composition for oral administration, any conventional pharmaceutical medium can be used. For example, when preparing oral liquid preparations, such as suspensions, elixirs, and solutions, water, ethylene glycol, oils, alcohols, flavoring agents, preservatives, coloring agents, etc. can be used. When preparing oral solid preparations, such as powders, hard capsules, soft capsules, and tablets, starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc. can be used. Solid oral preparations are preferred over liquid preparations.

[0049] Because of their ease of administration, tablets and capsules are the most advantageous oral dosage unit forms, in which case solid pharmaceutical carriers are obviously used. If necessary, tablets can be coated by standard aqueous or non-aqueous techniques. Such compositions and preparations should contain at least 0.1% of the active ingredient. Of course, the percentage of the active ingredient in these compositions can be varied, and the percentage can easily vary from about 2% to about 60% of the unit weight. The active ingredient can be administered intranasally, for example, as drops or spray.

[0050] The tablets, pills, capsules, etc. may contain binders (e.g., tragacanth, acacia, corn starch, or gelatin), excipients (e.g., dicalcium phosphate), disintegrating agents (e.g., corn starch, potato starch, alginic acid), lubricants (e.g., magnesium stearate), and sweeteners (e.g., sucrose, lactose, or saccharin). When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier (e.g., fatty oil).

[0051] Various other materials can be used to coat or otherwise modify the physical form of the dosage units. For example, tablets can be coated with shellac, sugar, or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetener, methylparaben or propylparaben as a preservative, a dye, and a flavoring (such as cherry or orange flavor).

[0052] Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of the present invention.

[0053] The amorphous substance of the present invention may be administered parenterally. A solution or suspension of these active substances can be prepared in water by appropriately mixing with a surfactant (e.g., hydroxypropyl cellulose). Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, and oils. Under normal conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0054] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the pharmaceutical form must be sterile and fluid so that it can be easily injected. It must be stable under the conditions of manufacture and storage and must be preserved in a condition that protects against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

[0055] Any suitable administration method can be used to provide an effective amount of the amorphous material of the present invention to a mammal, particularly a human, including oral, rectal, topical, parenteral, ocular, pulmonary, nasal, etc. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, emulsions, ointments, aerosols, etc.

[0056] The therapeutically effective amount of the amorphous, pharmaceutical composition, or combination thereof of the present invention will depend on the species, weight, age, and individual condition of the subject being treated, and the disorder or disease being treated or its severity. A physician, clinician, or veterinarian of ordinary skill can readily determine the effective amount of each of the active ingredients required to prevent, treat, or inhibit the progression of the disorder or disease.

[0057] When using the compounds of the present invention or their amorphous forms to treat or prevent diseases mediated by FXR according to the present invention, satisfactory results are generally obtained when the compounds of the present invention or their amorphous forms are administered at a daily dose of about 0.1 milligrams to about 100 milligrams per kilogram of animal body weight, preferably as a single dose, 2 to 6 divided doses per day, or in sustained-release form. For most large mammals, the total daily dose is about 1.0 milligrams to about 1000 milligrams, preferably about 1 milligram to about 50 milligrams. For a 70-kilogram adult, the total daily dose is generally about 7 milligrams to about 350 milligrams. This dosage regimen can be adjusted to obtain optimal therapeutic effects.

[0058] The amorphous substance or pharmaceutical composition thereof according to the present invention can prevent, treat, cure or alleviate diseases mediated by FXR in a patient, particularly 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 damage, gallstones, liver cirrhosis, hepatitis B, skin and gland diseases, alcoholic beverages, and the like. It can effectively treat liver cirrhosis caused by steroids, cystic fibrosis, biliary atresia, cholelithiasis, hepatic 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 tumor disease, and liver tumor disease. [Brief explanation of the drawings]

[0059] [Figure 1] 1 is an X-ray powder diffraction (XRPD) pattern of the amorphous compound represented by formula (I) prepared by the method of Example 1 of the present invention. [Figure 2] 1 is a differential scanning calorimetry (DSC) pattern of the amorphous compound represented by formula (I) prepared by the method of Example 1 of the present invention. [Figure 3] FIG. 1 is a thermogravimetric analysis (TGA) diagram of the amorphous compound represented by formula (I) prepared by the method of Example 1 of the present invention. [Figure 4] 1 shows the X-ray powder diffraction pattern of amorphous compound of formula (I) during the stability test at high temperature according to the method of Example 3 (1) of the present invention. [Figure 5] 1 shows the X-ray powder diffraction pattern of the amorphous compound of formula (I) during the stability test under high humidity conditions according to the method of Example 3(2) of the present invention. [Figure 6] 1 shows the X-ray powder diffraction pattern of amorphous compound of formula (I) during the stability test under light irradiation conditions according to method (3) of Example 3 of the present invention. [Figure 7] 1 shows the dynamic moisture sorption (DVS) pattern of the amorphous compound represented by formula (I) prepared by the method of Example 1 of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0060] The present invention will be further described below by way of examples, but the present invention is not limited to the scope of the above examples.

[0061] The X-ray powder diffraction analysis method used in this invention is as follows: X-ray powder diffraction patterns are obtained using Cu-Kα radiation (45 kV, 40 mA) on an Empyrean diffractometer. A thin layer is formed from the powdered sample on a single-crystal silicon sample holder, placed on a rotating sample stage, and analyzed within a range of 3° to 60° with a 0.0167° step size. Data are collected using Data Collector software, processed using High Score Plus software, and read using Data Viewer software.

[0062] The differential scanning calorimetry (DSC) analysis method used in this invention is to perform differential scanning calorimetry using a TAQ2000 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 cover, and heated at 10°C / min using a linear heating device from room temperature to approximately 300°C. The DSC cell is purged with dry nitrogen during use.

[0063] The thermogravimetric analysis (TGA) analysis method used in this invention is as follows: Thermogravimetric analysis is performed using a TAQ500 module 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 heated at 10°C / min using a linear heating device from room temperature to approximately 300°C. The DSC cell is purged with dry nitrogen during use.

[0064] Specific implementation methods For a specific synthesis method of the compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepin[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid represented by formula (I), see Example 9 of Chinese patent application CN107686486. [Example]

[0065] Example 1 Amorphous Form of Compound of Formula (I) 1. Amorphous Preparation At room temperature, 201.5 mg of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepin[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid was added to a 100 mL one-neck flask, followed by a homemade solution of sodium hydroxide (133.8 mg, 3.34 mmol) in water (30.0 mL). o Heat to 40°C. When the solid is completely dissolved, o The temperature was lowered to 100°C, and a self-prepared solution of dilute hydrochloric acid (4.74 mmol, 15.0 mL) was added dropwise. After the addition, the pH value was measured to be 2-3. The heating was stopped, the mixture was allowed to cool to room temperature, and the mixture was stirred for 3 hours to crystallize. The mixture was then suction filtered, and the filter cake was washed with water (3.0 mL × 3). o C for 6 hours to give a white solid (1.41 g, 92.2%).

[0066] 2. Identification of amorphous materials (1) X-ray powder diffraction (XRPD) analysis by Empyrean Identification: Cu-Kα radiation was used, and the X-ray powder diffraction pattern is substantially as shown in FIG.

[0067] (2) Analysis and identification of differential scanning calorimetry (DSC) by TAQ2000: the scanning rate was 10°C / min, and the amorphous material had a glass transition temperature of 92.26°C with a tolerance of ±3°C. The differential scanning calorimetry pattern of the amorphous material prepared by the method of this example was substantially as shown in Figure 2.

[0068] (3) Analysis and identification of thermogravimetric analysis (TGA) using TAQ500: the heating rate was 10°C / min, the weight loss was 0.409%, and the tolerance was ±0.1%. The thermogravimetric analysis diagram of the amorphous material prepared by the method of this example is substantially as shown in Figure 3.

[0069] Example 2 Pharmacokinetic study of the amorphous material according to the present invention Amorphous form of the compound represented by formula (I) according to the present invention (a compound called 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepin[3,2-b]pyridine-11,1'-cyclopropane]-7-carboxylic acid) was filled into capsules and orally administered.

[0070] Male Beagle dogs weighing 8-12 kg were orally administered 5 mg / kg of test substance, with three animals per group. Blood samples were collected at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 24 h post-administration. A standard curve with an appropriate range was established based on the sample concentrations, and the test substance concentrations in plasma samples were measured and quantitatively analyzed using an AB SCIEX API4000 LC-MS / MS in MRM mode. Based on the drug concentration-time curves, pharmacokinetic parameters were calculated using a non-compartmental model with WinNonLin 6.3 software. The test results are shown in Table 1.

[0071] Table 1. Pharmacokinetic study data of the amorphous substance according to the present invention [Table 2] Test conclusion: As can be seen from Table 1, the amorphous substance according to the present invention has a high exposure in the body of Beagle dogs and has good pharmacokinetic properties.

[0072] Example 3 Stability test of the amorphous material according to the present invention (1) High-temperature test: An appropriate amount of sample was placed in a flat weighing bottle, spread into a thin layer of 5 mm or less, and left at 60°C for 30 days. Samples were taken on the 5th, 10th, and 30th days to observe the color change of the sample. The purity of the sample was measured by HPLC, and the structure was analyzed by X-ray powder diffraction. The changes in the X-ray powder diffraction pattern after the high-temperature test were essentially as shown in Figure 4.

[0073] (2) High humidity test: An appropriate amount of sample was placed in a flat weighing bottle, spread into a thin layer of 5 mm or less, and left to stand at 25°C and RH 90%±5% for 30 days. Samples were taken on the 5th, 10th, and 30th days to observe the color change of the sample, measure the purity of the sample by HPLC, and analyze the structure by X-ray powder diffraction. The changes in the X-ray powder diffraction pattern after the high humidity test were essentially as shown in Figure 5.

[0074] (3) Light irradiation test: Put an appropriate amount of test sample into a flat weighing bottle, spread it into a thin layer with a thickness of 5 mm or less, and place it in a light irradiation test box. With the box open, expose it to an illuminance of 4500 ± 500 lx and ultraviolet light ≥ 0.7 w / m 2 The sample was left to stand for 30 days under the above conditions, and samples were taken on the 5th, 13th, and 30th days. The color change of the sample was observed, the purity of the sample was measured by HPLC, and the structure was analyzed by X-ray powder diffraction. The changes in the X-ray powder diffraction pattern during the light irradiation test were substantially as shown in Figure 6.

[0075] Table 2 shows the changes in appearance and chemical purity of the test samples during the stability test.

[0076] Table 2. Stability test of amorphous materials [Table 3]

[0077] Test conclusion: Under conditions of high temperature, high humidity and light irradiation, the appearance, chemical purity and crystalline form of the amorphous material of the present invention do not change obviously, and it is highly stable and suitable for medical use.

[0078] Example 4 Hygroscopicity test of amorphous material according to the present invention The hygroscopicity of an appropriate amount of sample was measured using a dynamic moisture sorption apparatus. The test results are shown in Figure 7. The test results showed that the amorphous material of the present invention exhibited a weight gain of less than 2% upon equilibrium at a relative humidity of 80%, which is low hygroscopicity according to the standard definition of hygroscopic mass gain. In other words, the amorphous material of the present invention is not easily deliquesced by high humidity.

[0079] The above is a basic explanation of the concept of the present invention, and any equivalent transformations made based on the technical means of the present invention should fall within the protection scope of the present invention.

[0080] In the description herein, the references such as "one embodiment," "some embodiments," "examples," "specific examples," or "several examples" mean that the specific features, structures, materials, or characteristics described in combination with the embodiment or example are included in at least one embodiment or example of the present invention. In the description herein, the exemplary expressions of the above terms are not necessarily limited to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be appropriately combined in any one or more embodiments or examples. Furthermore, unless mutually inconsistent, a person skilled in the art may combine or combine different embodiments or examples and features of different embodiments or examples described herein.

[0081] Although the embodiments of the present invention have been illustrated and described above, it should be understood that the above embodiments are merely illustrative and should not be construed as limiting the present invention, and that those skilled in the art may make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention.

Claims

1. An amorphous compound represented by formula (I), 【Table 1】 Amorphous having an X-ray powder diffraction pattern as shown in 【Chemical 1】

2. An amorphous form of the compound of formula (I), having a glass transition temperature of 92.26°C ± 3°C. 【Chemistry 2】 【Request 3】 【Table 2】 3. The amorphous material of claim 2, having a differential scanning calorimetry pattern as shown in

4. A pharmaceutical composition comprising the amorphous material according to any one of claims 1 to 3 and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, or combination thereof.

5. Use of an amorphous substance according to any one of claims 1 to 3 or a pharmaceutical composition according to claim 4 in the manufacture of a medicament for preventing, treating or ameliorating a disease mediated by FXR in a patient.

6. The use according to claim 5, wherein the FXR-mediated disease is a cardio-cerebrovascular disease, a lipid metabolism disorder-related disease, a metabolic syndrome, a hyperproliferative disease, a fibrosis, an inflammatory disease or a disease related to the liver and gallbladder.

7. The cardiocerebrovascular disease is atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke, or thrombosis; The metabolic syndrome is insulin resistance, hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, 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 complication of diabetes and an abnormally high body mass index; The hyperproliferative diseases are hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma, breast cancer, membranous adenocarcinoma, Barrett's esophagus cancer and other forms of gastrointestinal or hepatic neoplastic diseases; 7. The use according to claim 6, wherein the fibrosis, inflammatory disease or disease related to the liver and gallbladder is non-alcoholic fatty liver, non-alcoholic steatohepatitis, cholestasis, hepatic fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial cholestasis, cystic fibrosis, drug-induced bile duct damage, gallstones, cirrhosis, hepatitis B, glandular skin disease, alcoholic cirrhosis, biliary atresia, cholelithiasis, colitis, neonatal jaundice, kernicterus or intestinal bacterial overgrowth.

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