WO2024140868A1 - Crystal form of tricyclic compound and use thereof - Google Patents

Abstract

Provided are a crystal form of a tricyclic compound and a use thereof. Specifically, provided are a crystal form of a compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazole [3,2-b]pyridine-7-carboxylic acid, a pharmaceutical composition comprising the crystal form, and a use thereof in the preparation of a drug for preventing, treating, or alleviating FXR-mediated diseases in patients.

Description

Crystal form of tricyclic compound and its use Technical Field

The present invention belongs to the field of medical technology, and relates to a crystal form of a tricyclic compound and a use thereof, and specifically relates to a crystal form of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazepine[3,2-b]pyridine-7-carboxylic acid and a pharmaceutical composition comprising the crystal form, and further relates to their use in preparing a drug for treating diseases mediated by FXR.

Background technique

Farnesoid X receptor (FXR) is a member of the nuclear hormone receptor superfamily and is primarily expressed 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 retinoic acid X receptor (RXR), binding to response elements in target gene promoters to regulate gene transcription. The FXR-RXR heterodimer binds with the highest affinity to the inverted repeat-1 (IR-1) response element, where the hexamer that binds the consensus receptor is separated by one nucleotide. FXR can be activated by bile acids, the end product of cholesterol metabolism (Makishima et al. Science (1999), 284:1362-1365; Parks et al. Science (1999), 284:1365-1368; Wang et al. MoI. Cell. (1999), 3:543-553), and bile acids serve to inhibit the catabolism of cholesterol (Urizar et al., (2000) J. Biol. Chem. 275:39313-393170).

FXR is a key regulator of cholesterol homeostasis, triglyceride synthesis and lipogenesis (Crawley, Expert Opinion Ther. Patents (2010), 20:1047-1057). In addition to being a target for the treatment of dyslipidemia, obesity, vitamin D-related diseases, intestinal diseases, drug-induced side effects and hepatitis (Crawley, Expert Opinion Ther. Patents (2010), 20:1047-1057), FXR can also be used as a therapeutic target for hepatobiliary diseases, chronic hepatitis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cholestasis, liver fibrosis, cirrhosis, hepatitis B, metabolic diseases, lipid metabolism diseases, carbohydrate metabolism diseases, cardiovascular metabolic diseases, atherosclerosis, type 2 diabetes and diabetic complications (Frank G. Schaap et al., Journal of Medicinal Chemistry (2005), 48:5383-5402).

Patent applications WO 2016127924A1 and CN 105884758A disclose tricyclic compounds that can be used as FXR activity regulators, as well as preparation methods and applications thereof, wherein Example 3 is specifically disclosed, namely, compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (compound shown in formula (I)).

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 crystalline forms of the same drug may differ significantly in appearance, solubility, melting point, dissolution, bioavailability, etc., and may also affect the stability and bioavailability of the drug. In order to better control the quality of the drug and meet the requirements of preparation, production, transportation, storage, etc., it is necessary to study the crystal form of the compound represented by formula (I) in order to find a crystal form with good properties.

Summary of the invention

The present invention provides a crystalline form of a compound represented by formula (I). Specifically, the crystalline form of a compound represented by formula (I) provided by the present invention can significantly improve the stability and pharmacokinetic properties of the compound, thereby having better drugability.

Specifically, the present invention relates to a crystalline form of a compound represented by formula (I), and a pharmaceutical composition comprising the crystalline form, and also relates to their use in preparing a drug for preventing, treating or alleviating a disease mediated by FXR in a patient.

In one aspect, the present invention provides a crystalline form of a compound represented by formula (I);

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the X-ray powder diffraction pattern of the crystalline form II comprises the following diffraction peaks at 2θ angles: 16.97°±0.2°, 17.18°±0.2°, 21.31°±0.2°, 25.29°±0.2°, 26.36°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the X-ray powder diffraction pattern of the crystalline form II comprises the following diffraction peaks at 2θ angles: 11.36°±0.2°, 13.45°±0.2°, 16.97°±0.2°, 17.18°±0.2°, 20.77°±0.2°, 21.08°±0.2°, 21.31°±0.2°, 23.56°±0.2°, 24.31°±0.2°, 24.52°±0.2°, 25.29°±0.2°, 26.36°±0.2°, 26.67°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the X-ray powder diffraction pattern of the crystalline form II comprises the following diffraction peaks at 2θ angles: 5.65°±0.2°, 9.63°±0.2°, 11.36°±0.2°, 11.84°±0.2°, 12.11°±0.2°, 12.71°±0.2°, 13.45°±0.2°, 14.69°±0.2°, 14.96°±0.2°, 16.42°±0.2°, 16.9 7°±0.2°,17.18°±0.2°,17.77°±0.2°,18.26°±0.2°,18.89°±0.2°,19.30°±0.2°,20.11°±0.2°,20.77°±0.2°,21.08°±0.2°,21.31°±0.2°,21.90°±0.2°,22.81°±0.2°,23.56°±0.2°,24.31°±0.2°,24.52°±0.2°,25.29°±0.2°,26. 36°±0.2°,26.67°±0.2°,27.03°±0.2°,27.69°±0.2°,27.97°±0.2°,28.33°±0.2°,28.67°±0.2°,29.11°±0.2°,29.91°±0.2°,30.29°±0.2°,30.62°±0.2°,31.54°±0.2°,31.81°±0.2°,32.35°±0.2°,33.15°±0.2°,33.79°±0.2°,3 : 4.29°±0.2°,35.10°±0.2°,35.75°±0.2°,36.64°±0.2°,37.99°±0.2°,38.82°±0.2°,39.67°±0.2°,40.78°±0.2°,43.11°±0.2°,43.90°±0.2°,44.95°±0.2°,45.90°±0.2°,48.79°±0.2°,51.65°±0.2°,52.76°±0.2°,53.79°±0.2°.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the crystalline form II has an X-ray powder diffraction pattern substantially as shown in FIG. 1 .

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the differential scanning calorimetry diagram of the crystalline form II comprises endothermic peaks at 125.84°C±3°C and 198.41°C±3°C.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the crystalline form II has a differential scanning calorimetry diagram substantially as shown in FIG. 2 .

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the crystalline form II loses weight by 7.233% in the range of 70°C-160°C, with an error tolerance of ±0.1%.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form II, characterized in that the crystalline form II has a thermogravimetric analysis diagram substantially as shown in FIG. 3 .

In some embodiments, the crystalline form of the compound represented by formula (I) described in the present invention is crystalline form III, characterized in that the X-ray powder diffraction pattern of the crystalline form III comprises the following diffraction peaks at 2θ angles: 17.17°±0.2°, 20.92°±0.2°, 24.38°±0.2°, 25.19°±0.2°, 26.21°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) described in the present invention is crystalline form III, characterized in that the X-ray powder diffraction pattern of the crystalline form III comprises the following diffraction peaks at 2θ angles: 9.59°±0.2°, 17.17°±0.2°, 20.92°±0.2°, 21.12°±0.2°, 23.51°±0.2°, 24.38°±0.2°, 24.67°±0.2°, 25.19°±0.2°, 25.98°±0.2°, 26.21°±0.2°, 26.83°±0.2°, 28.59°±0.2°, 28.92°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form III, characterized in that the X-ray powder diffraction pattern of the crystalline form III comprises the following diffraction peaks at 2θ angles: 5.70°±0.2°, 9.01°±0.2°, 9.59°±0.2°, 10.95°±0.2°, 11.43°±0.2°, 11.95°±0.2°, 12.15°±0.2°, 13.55°±0.2°, 14.18°±0.2°, 14.82°±0.2°, 15. 09°±0.2°,15.56°±0.2°,16.50°±0.2°,17.17°±0.2°,17.74°±0.2°,17.88°±0.2°,18.37°±0.2°,19.03°±0.2°,19.40°±0.2°,20.13°±0.2°,20.92°±0.2°,21.12°±0.2°,21.48°±0.2°,21.81°±0.2°,22.90°±0.2°,23.51°±0.2°,24 .38°±0.2°,24.67°±0.2°,25.19°±0.2°,25.98°±0.2°,26.21°±0.2°,26.83°±0.2°,27.21°±0.2°,27.50°±0.2°,28.01°±0.2°,28.59°±0.2°,28.92°±0.2°,29.45°±0.2°,29.84°±0.2°,30.79°±0.2°,31.37°±0.2°,31.74°±0.2°,3 : 2.47°±0.2°,33.58°±0.2°,34.08°±0.2°,35.83°±0.2°,36.47°±0.2°,38.11°±0.2°,38.93°±0.2°,40.60°±0.2°,42.59°±0.2°,42.79°±0.2°,43.63°±0.2°,45.20°±0.2°,46.52°±0.2°,48.74°±0.2°,51.65°±0.2°,53.46°±0.2°.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form III, characterized in that the crystalline form III has an X-ray powder diffraction pattern substantially as shown in FIG. 4 .

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form III, characterized in that the differential scanning calorimetry diagram of the crystalline form III comprises endothermic peaks at 104.93°C±3°C and 199.52°C±3°C.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form III, characterized in that the crystalline form III has a differential scanning calorimetry diagram substantially as shown in FIG. 5 .

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form III, characterized in that the crystalline form III loses weight by 4.383% in the range of 70°C-160°C, with an error tolerance of ±0.1%.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form III, characterized in that the crystalline form III has a thermogravimetric analysis diagram substantially as shown in FIG. 6 .

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 13.92°±0.2°, 14.93°±0.2°, 19.48°±0.2°, 23.23°±0.2°, 23.55°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0.2°, 19.48°±0.2°, 20.60°±0.2°, 22.19°±0.2°, 22.66°±0.2°, 23.23°±0.2°, 23.55°±0.2°, 24.06°±0.2°, 26.42°±0.2°, 26.75°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0.2°, 17.97°±0.2°, 18.57°±0.2°, 18.86°±0.2°, 19.48°±0.2°, 20.60°±0.2°, 21.72°± 0.2°,22.19°±0.2°,22.66°±0.2°,23.23°±0.2°,23.55°±0.2°,24.06°±0.2°,25.04°±0.2°,25.69°±0.2°,26.42°±0.2°,26.75°±0.2°,27.93°±0.2°,30.25°±0.2°,31.47°±0.2°,32.19°±0.2°,35.83°±0.2°,36.61°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 10.57°±0.2°, 10.90°±0.2°, 11.87°±0.2°, 12.91°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0.2°, 16.35°±0.2° ,17.10°±0.2°,17.97°±0.2°,18.57°±0.2°,18.86°±0.2°,19.48°±0.2°,20.60°±0.2°,21.08°±0.2°,21.72°±0.2°,22.19°±0.2°,22.66°±0.2°,23.23°±0.2°,23.55°±0.2°,24.06°±0.2°,25.04°±0.2°,25.69°± 0.2°,26.42°±0.2°,26.75°±0.2°,27.14°±0.2°,27.93°±0.2°,28.47°±0.2°,28.71°±0.2°,29.06°±0.2°,29.39°±0.2°,30.25°±0.2°,30.64°±0.2°,31.47°±0.2°,31.82°±0.2°,32.19°±0.2°,32.84°±0.2°,33. : 95°±0.2°,34.51°±0.2°,35.43°±0.2°,35.83°±0.2°,36.28°±0.2°,36.61°±0.2°,37.12°±0.2°,37.89°±0.2°,38.82°±0.2°,40.68°±0.2°,41.10°±0.2°,41.53°±0.2°,43.14°±0.2°,47.35°±0.2°,51.15°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 10.57°±0.2°, 10.90°±0.2°, 11.87°±0.2°, 12.91°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0.2°, 16.35°±0.2°, 17.10°±0.2°, 17.97°±0.2°, 18.57°±0.2°, 18.86°±0.2°, 19.48°±0.2°, 20.60 °±0.2°,21.08°±0.2°,21.72°±0.2°,22.19°±0.2°,22.66°±0.2°,23.23°±0.2°,23.55°±0.2°,24.06°±0.2°,25.04°±0.2°,25.69°±0.2°,26.42°±0.2°,26.75°±0.2°,27.14°±0.2°,27.93°±0.2°,28.47°±0.2°,28.71°±0.2°,29.06°±0.2°,29.39°±0.2°,30.25°±0.2°,30.64°±0.2°,31.47°± 0.2°,31.82°±0.2°,32.19°±0.2°,32.84°±0.2°,33.95°±0.2°,34.51°±0.2°,35.43°±0.2°,35.83°±0.2°,36.28°±0.2°,36.61°±0.2°,37.12°±0.2°,37.89°±0.2°,38.82°±0.2°,39.64°±0.2°,40.16°±0. 2°,40.68°±0.2°,41.10°±0.2°,41.53°±0.2°,43.14°±0.2°,44.15°±0.2°,45.71°±0.2°,46.33°±0.2°,46.83°±0.2°,47.35°±0.2°,51.15°±0.2°,52.63°±0.2°,54.22°±0.2°,55.16°±0.2°,57.64°±0.2°.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the crystalline form VII has an X-ray powder diffraction pattern substantially as shown in Figure 7.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the differential scanning calorimetry diagram of the crystalline form VII comprises endothermic peaks at 177.53°C±3°C and 199.42°C±3°C and an exothermic peak at 179.32°C±3°C.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the crystalline form VII has a differential scanning calorimetry diagram substantially as shown in Figure 8.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the crystalline form VII loses weight by 0.3968% in the range of 70°C-160°C, with an error tolerance of ±0.1%.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VII, characterized in that the crystalline form VII has a thermogravimetric analysis diagram substantially as shown in FIG. 9 .

In some embodiments, the crystalline form of the compound represented by formula (I) described in the present invention is crystalline form VIII, characterized in that the X-ray powder diffraction pattern of the crystalline form VIII comprises the following diffraction peaks at 2θ angles: 17.24°±0.2°, 20.92°±0.2°, 24.33°±0.2°, 25.19°±0.2°, 26.31°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) described in the present invention is crystalline form VIII, characterized in that the X-ray powder diffraction pattern of the crystalline form VIII comprises the following diffraction peaks at 2θ angles: 9.60°±0.2°, 12.12°±0.2°, 17.24°±0.2°, 20.92°±0.2°, 23.65°±0.2°, 24.33°±0.2°, 24.60°±0.2°, 25.19°±0.2°, 26.31°±0.2°, 27.55°±0.2°, 28.51°±0.2°, 29.01°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VIII, characterized in that the X-ray powder diffraction pattern of the crystalline form VIII comprises the following diffraction peaks at 2θ angles: 5.70°±0.2°, 9.60°±0.2°, 11.34°±0.2°, 11.82°±0.2°, 12.12°±0.2°, 12.71°±0.2°, 13.35°±0.2°, 14.75°±0.2°, 14.89°±0.2°, 16.48°±0.2°, 17.24°±0.2°, 17.75°±0.2° ,18.08°±0.2°,19.00°±0.2°,19.24°±0.2°,19.98°±0.2°,20.92°±0.2°,21.25°±0.2°,21.87°±0.2°,22.73°±0.2°,23.65°±0.2°,24.33°±0.2°,24.60°±0.2°,25.19°±0.2°,25.58°±0.2°,26.31°±0.2°,26.54°±0.2°,26.91°±0.2°,27.55°±0.2°,28.09°±0. 2°,28.51°±0.2°,28.79°±0.2°,29.01°±0.2°,29.30°±0.2°,29.96°±0.2°,30.85°±0.2°,31.47°±0.2°,31.80°±0.2°,32.17°±0.2°,32.81°±0.2°,33.66°±0.2°,34.15°±0.2°,34.85°±0.2°,35.63°±0.2°,35.94°±0.2°,36.54°±0.2°,36.91°±0.2°,38.00°± 0.2°,39.08°±0.2°,40.96°±0.2°,42.55°±0.2°,42.92°±0.2°,43.63°±0.2°,44.88°±0.2°,46.17°±0.2°,46.47°±0.2°,48.68°±0.2°,50.02°±0.2°,51.23°±0.2°,51.77°±0.2°,52.40°±0.2°,53.28°±0.2°,53.72°±0.2°,54.70°±0.2°,56.96°±0.2°,58.34° ±0.2°.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VIII, characterized in that the crystalline form VIII has an X-ray powder diffraction pattern substantially as shown in FIG. 10 .

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VIII, characterized in that the differential scanning calorimetry diagram of the crystalline form VIII comprises endothermic peaks at 116.13°C±3°C, 191.72°C±3°C and 199.49°C±3°C.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VIII, characterized in that the crystalline form VIII has a differential scanning calorimetry diagram substantially as shown in FIG. 11 .

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VIII, characterized in that the crystalline form VIII loses 2.081% of its weight in the range of 70°C-160°C, with an error tolerance of ±0.1%.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form VIII, characterized in that the crystalline form VIII has a thermogravimetric analysis diagram substantially as shown in FIG. 12 .

In some embodiments, the crystalline form of the compound represented by formula (I) described in the present invention is crystalline form IX, characterized in that the X-ray powder diffraction pattern of the crystalline form IX comprises the following diffraction peaks at 2θ angles: 15.92°±0.2°, 21.17°±0.2°, 21.82°±0.2°, 22.44°±0.2°, 23.49°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) described in the present invention is crystalline form IX, characterized in that the X-ray powder diffraction pattern of the crystalline form IX comprises the following diffraction peaks at 2θ angles: 5.30°±0.2°, 15.92°±0.2°, 17.18°±0.2°, 20.23°±0.2°, 21.17°±0.2°, 21.27°±0.2°, 21.82°±0.2°, 22.08°±0.2°, 22.20°±0.2°, 22.44°±0.2°, 23.39°±0.2°, 23.49°±0.2°, 26.46°±0.2°.

In other embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form IX, characterized in that the X-ray powder diffraction pattern of the crystalline form IX comprises the following diffraction peaks at 2θ angles: 5.30°±0.2°, 10.33°±0.2°, 10.59°±0.2°, 11.12°±0.2°, 11.95°±0.2°, 12.96°±0.2°, 13.16°±0.2°, 13.31°±0.2°, 13.48°±0.2°, 15.92°±0.2°, 16.45°±0.2°, 17.18°±0.2°, 19.52°±0.2°, 19.90°±0.2°, 20.23°±0.2°. 2°,20.68°±0.2°,21.17°±0.2°,21.27°±0.2°,21.82°±0.2°,22.08°±0.2°,22.20°±0.2°,22.44°±0.2°,23.39°±0.2°,23.49°±0.2°,24.02°±0.2° ,24.26°±0.2°,24.89°±0.2°,25.26°±0.2°,25.90°±0.2°,26.30°±0.2°,26.46°±0.2°,26.75°±0.2°,27.16°±0.2°,27.55°±0.2°,28.72°±0.2°,29 .34°±0.2°,29.81°±0.2°,30.22°±0.2°,30.42°±0.2°,31.24°±0.2°,32.10°±0.2°,32.68°±0.2°,32.92°±0.2°,33.25°±0.2°,33.59°±0.2°,34.0 5°±0.2°,34.81°±0.2°,35.20°±0.2°,35.82°±0.2°,36.13°±0.2°,36.36°±0.2°,37.59°±0.2°,38.27°±0.2°,39.05°±0.2°,39.57°±0.2°,40.21°± 0.2°,41.78°±0.2°,42.15°±0.2°,42.63°±0.2°,43.21°±0.2°,43.94°±0.2°,44.92°±0.2°,45.94°±0.2°,46.83°±0.2°,47.44°±0.2°,49.08°±0. 2°,49.65°±0.2°,50.40°±0.2°,50.71°±0.2°,52.27°±0.2°,53.05°±0.2°,53.89°±0.2°,54.66°±0.2°,55.83°±0.2°,57.68°±0.2°,59.39°±0.2°.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form IX, characterized in that the crystalline form IX has An X-ray powder diffraction pattern substantially as shown in Figure 13.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form IX, characterized in that the differential scanning calorimetry diagram of the crystalline form IX comprises endothermic peaks at 111.76°C±3°C, 193.61°C±3°C and 199.64°C±3°C.

In some embodiments, the crystalline form of the compound represented by formula (I) of the present invention is crystalline form IX, characterized in that the crystalline form IX has a differential scanning calorimetry diagram substantially as shown in Figure 14.

In one aspect, the present invention relates to a pharmaceutical composition comprising the crystal form of the present invention and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant or a combination thereof.

On the other hand, the present invention also relates to the use of the crystalline form of the compound of formula (I) or the pharmaceutical composition in the preparation of a drug, wherein the drug is used to prevent, treat or alleviate a disease mediated by FXR in a patient; further, the use comprises administering an effective therapeutic dose of the crystalline form or the pharmaceutical composition of the present invention to a human or animal.

In some embodiments, the FXR-mediated disease of the present invention is a cardiovascular and cerebrovascular disease, a disease associated with dyslipidemia, a metabolic syndrome, a hyperproliferative disease, fibrosis, an inflammatory disease, or a hepatobiliary-related disease.

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

In other embodiments, the metabolic syndrome described in the present invention is insulin resistance, hyperglycemia, hyperinsulinemia, elevated levels of fatty acids or glycerol 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.

In other embodiments, the hyperproliferative disease described herein is hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma, breast cancer, pancreatic cancer, Bartter's esophagus cancer, and other forms of gastrointestinal or liver neoplastic diseases.

In other embodiments, the fibrosis, inflammatory disease or hepatobiliary-related disease described in the present invention is 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 damage, gallstones, cirrhosis, hepatitis B, sebaceous gland disease, alcohol-induced cirrhosis, biliary obstruction, cholelithiasis, colitis, neonatal jaundice, nuclear jaundice or intestinal bacterial overgrowth.

In one aspect, the present invention relates to a method for preventing, treating or alleviating a disease mediated by FXR in a patient, comprising administering a pharmaceutically acceptable effective dose of the crystalline form or the pharmaceutical composition of the present invention to the patient.

On the other hand, the present invention also relates to a method for preparing the crystalline form of the compound represented by formula (I).

The solvent used in the method for preparing the crystal form described in the present invention is not particularly limited, and any solvent that can dissolve the starting material to a certain extent and does not affect its properties is included in the present invention. In addition, many similar modifications, equivalent replacements, or solvents, solvent combinations, and different proportions of solvent combinations equivalent to those described in the present invention in the art are considered to be included in the scope of the present invention. The present invention provides preferred solvents used in each reaction step.

The preparation experiment of the crystal form described in the present invention will be described in detail in the embodiment section. At the same time, the present invention provides activity test experiments of the crystal form, such as pharmacokinetic experiments, stability experiments and hygroscopic experiments. Experimental studies have found that the properties (such as stability) of individual crystal forms of the prior art and the compound shown in formula (I) are not good, for example, crystal transformation occurs under heating or high temperature conditions. Further studies have found that compared with the prior art or other crystal forms, the crystal form VII of the compound shown in formula (I) described in the present invention has better biological activity and high stability, and is suitable for pharmaceutical use. Specifically, the crystal form VII described in the present invention has more excellent pharmacokinetic properties and higher stability. For example, the crystal form VII has a higher exposure amount and a higher blood drug concentration.

Definitions and general terms

Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those of ordinary skill in the art to which the present invention belongs. All patents and publications to which the present invention relates are incorporated herein by reference in their entirety. Although any methods and materials similar or identical to those described in the present invention may be used in the practice or testing of the present invention, preferred methods, apparatus and materials are described in the present invention.

"Crystal form" or "crystalline form" refers to a solid having a highly regular chemical structure, including, but not limited to, single-component or multi-component crystals, and/or polymorphs, solvates, hydrates, inclusion compounds, co-crystals, salts, solvates of salts, hydrates of salts of compounds. The crystalline form of a substance can be obtained by many methods known in the art. Such methods include, but are not limited to, melt crystallization, melt cooling, solvent crystallization, crystallization in a confined space, for example, in a nanopore or capillary, crystallization on a surface or template, for example, on a polymer, crystallization in the presence of an additive such as a co-crystallization countermolecule, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, reactive crystallization, anti-solvent addition, grinding and solvent drop grinding, etc.

"Solvent" refers to a substance (typically a liquid) that is capable of completely or partially dissolving another substance (typically a solid). Solvents useful 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, 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, and the like.

"Anti-solvent" refers to a fluid that promotes precipitation of a product (or a product precursor) from a solvent. The anti-solvent may include a cold gas, or a fluid that promotes precipitation by a 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 from the solvent.

"Solvate" refers to a compound having a solvent on the surface, in the crystal lattice, or on the surface and in the crystal lattice, and the solvent may 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, methylpyrrolidone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, and mixtures thereof, etc. A specific example of a solvate is a hydrate, in which the solvent on the surface, in the crystal lattice, or on the surface and in the crystal lattice is water. On the surface, in the crystal lattice, or on the surface and in the crystal lattice of a substance, a hydrate may or may not have solvents other than water. Other solvents.

The crystal form can be identified by a variety of technical means, 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, solution calorimetry, scanning electron microscopy (SEM), quantitative analysis, solubility and dissolution rate, etc.

X-ray powder diffraction (XRPD) can detect information such as changes in crystal forms, crystallinity, and crystal structure states, and is a common means of identifying crystal forms. The peak position of the XRPD spectrum depends mainly on the structure of the crystal form, is relatively insensitive to experimental details, and its relative peak height depends on many factors related to sample preparation and instrument geometry. Therefore, in some embodiments, the crystal form of the present invention is characterized by an XRPD pattern with certain peak positions, which is substantially as shown in the XRPD pattern provided in the accompanying drawings of the present invention. At the same time, the measurement of 2θ of the XRPD spectrum may have experimental errors, and the measurement of 2θ of the XRPD spectrum may be slightly different between different instruments and different samples, so the value of the 2θ cannot be regarded as absolute. According to the instrument conditions used in this experiment, there is an error tolerance of ±0.2 ^o for the diffraction peak.

Differential scanning calorimetry (DSC) is a technique that measures the energy difference between a sample and an inert reference material (usually α-Al ₂ O ₃ ) as it changes with temperature by continuously heating or cooling under program control. The endothermic peak height of the DSC curve depends on many factors related to sample preparation and instrument geometry, while the peak position is relatively insensitive to experimental details. Therefore, in some embodiments, the crystal form of the present invention is characterized by a DSC graph with a characteristic peak position, which is substantially as shown in the DSC graph provided in the accompanying drawings of the present invention. At the same time, the DSC spectrum may have experimental errors, and the peak position and peak value of the DSC spectrum may vary slightly between different instruments and different samples, so the peak position or peak value of the DSC endothermic peak cannot be regarded as absolute. According to the instrument conditions used in this experiment, the endothermic peak has an error tolerance of ±3 ^° .

In the context of the present invention, 2θ values in X-ray powder diffraction patterns are given in degrees (°).

The term "substantially 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 99% of the peaks in the X-ray powder diffraction pattern, or the DSC pattern, or the Raman spectrum pattern, or the infrared spectrum pattern are shown in the pattern.

When referring to a spectrum and/or data appearing in a graph, a "peak" refers to a feature that can be identified by one skilled in the art and which cannot be attributed to background noise.

The present invention relates to various new crystalline forms of 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid, for example, form VII, which exist in a substantially pure crystalline form.

"Substantially pure" means that one crystalline form is substantially free of one or more other crystalline forms, that is, the purity of the crystalline form is at least 80%, or at least 85%, or at least 90%, or at least 93%, or at least 95%, 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 crystalline form contains other crystalline forms, and the percentage of the other crystalline forms in the total volume or total weight of the crystalline form is less than 20%, or less than 10%, or less than 5%, or less than 3%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01%.

"Substantially free" means that the percentage of one or more other crystalline forms in the total volume or total weight of the crystalline 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%.

The "relative intensity" (or "relative peak height") in the XRPD pattern refers to the first strongest peak among all the diffraction peaks in the X-ray powder diffraction pattern (XRPD). The ratio of the intensity of other peaks to the intensity of the first strongest peak when the intensity is 100%.

In the context of the present invention, when or whether the words "about" or "approximately" are used, it means within 10%, suitably within 5%, and especially within 1% of a given value or range. Alternatively, for those of ordinary skill in the art, the term "about" or "approximately" means within an acceptable standard error range of the mean. Whenever a number having 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% will be explicitly disclosed, where "+/-" means plus or minus.

In the present 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.

Pharmaceutical compositions, preparations, administration and uses of the crystal forms of the compounds of the present invention

The pharmaceutical composition of the present invention is characterized by comprising any crystalline form of the compound represented by formula (I) or any combination thereof and a pharmaceutically acceptable carrier, adjuvant, or excipient. The amount of the crystalline form of the compound in the pharmaceutical composition of the present invention is effective and detectable to treat or alleviate the disease mediated by FXR in the patient.

As described in the present invention, the pharmaceutically acceptable composition of the present invention further comprises a pharmaceutically acceptable carrier, adjuvant, or excipient, which, as used in the present invention, includes 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 specific target dosage form. As described in the following documents: 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, Comprehensively the contents of the documents here, it is shown that different carriers can be used for the preparation of pharmaceutically acceptable compositions and their known preparation methods. Except to the extent that any conventional carrier medium is incompatible with the compounds of the present invention or their crystalline forms, such as by producing any adverse biological effects or interacting in a deleterious manner with any other components of the pharmaceutically acceptable composition, their use is also contemplated by the present invention.

The crystal form of the present invention can be used as an active ingredient and uniformly combined in a mixture with a drug carrier according to conventional drug compounding technology. Depending on the dosage form required for administration, such as oral or parenteral (including intravenous), the carrier can be in a variety of forms. When preparing a composition for oral dosage form, any conventional drug medium can be used, for example, water, ethylene glycol, oil, alcohol, fragrance, preservative, colorant, etc. are used when preparing oral liquid medicaments such as suspensions, elixirs and solutions; or when preparing oral solid preparations such as powders, hard capsules, soft capsules and tablets, starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, adhesives, disintegrants, etc. are used, wherein solid oral preparations are more preferred than liquid medicaments.

Because tablets and capsules are easy to take, they represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously used. If desired, the tablets can be coated using standard aqueous or non-aqueous techniques. Such compositions and preparations should contain at least 0.1 percent of the active ingredient. Of course, the percentage of active ingredient in these compositions can be varied and can conveniently vary between about 2% and about 60% of the unit weight. The active ingredient can be administered intranasally, for example, in the form of drops or sprays.

The tablets, pills, capsules, etc. may also contain: a binder (such as gum tragacanth, gum arabic, corn starch or gelatin); an excipient (such as dicalcium phosphate); a disintegrant (such as corn starch, potato starch, alginic acid); a lubricant (such as magnesium stearate); and a sweetener (such as sucrose, lactose or saccharin). When the dosage unit form is a capsule, it may contain a liquid carrier (such as a fatty oil) in addition to the above-mentioned types of materials.

Various other materials may be present as coatings or to modify the appearance of the dosage unit. For example, tablets may be coated with shellac, sugar, or both. In addition to the active ingredient, a syrup or elixir may contain sucrose as a sweetener, methyl or propyl parabens as preservatives, dyes, and flavorings (e.g., cherry or orange flavors).

Also included within the scope of this invention are ophthalmic formulations, eye ointments, powders, solutions, and the like.

The crystalline forms of the present invention may also be administered parenterally. Solutions or suspensions of these active substances may be prepared in water by suitable mixing with a surfactant such as hydroxypropylcellulose. Dispersants may also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof, and in oils. Under normal conditions of storage and use, these preparations contain preservatives to prevent the growth of microorganisms.

The pharmaceutical form suitable for injection purposes includes sterile aqueous solution or dispersant and the sterile powder for instant preparation of sterile injectable solution or dispersant. In all cases, the pharmaceutical form must be sterile and must be a fluid in the form of easy injection. It must be stable under the conditions of manufacture and storage and must be preserved under the conditions of antimicrobial such as antibacterial and fungal contamination. Carrier can be a solvent or a dispersion medium, which contains, for example: water, ethanol, polyols (such as glycerol, propylene glycol and liquid polyethylene glycol), their applicable mixtures and vegetable oils.

Any suitable method of administration can be used to provide an effective dose of the crystalline form of the present invention to a mammal, especially a human. For example, oral, rectal, topical, parenteral, ocular, pulmonary, nasal and other administration methods can be used. Dosage forms include tablets, lozenges, dispersants, suspensions, solutions, capsules, emulsions, ointments, aerosols, etc. Preferably, the crystalline form of the present invention is administered orally.

The therapeutically effective dose of the crystalline form of the present invention, pharmaceutical composition or combination thereof depends on the species, body weight, age and individual condition of the individual, the disorder or disease to be treated or its severity. A physician, clinician or veterinarian of ordinary skill can easily determine the effective amount required for each active ingredient to prevent, treat the disorder or disease or inhibit the progression of the disorder or disease.

When using the compounds of the present invention or their crystalline forms to treat or prevent the FXR-mediated conditions described in the present invention, approximately satisfactory results are obtained when the compounds of the present invention or their crystalline forms are administered at a daily dose of about 0.1 mg to about 100 mg/kg of animal body weight, preferably in a single daily dose, or in 2 to 6 divided doses per day, or in a continuous release form. For most large mammals, the total daily dose is about 1.0 mg to about 1000 mg, preferably about 1 mg to about 50 mg. For a 70 kg adult, the total daily dose is generally 7 mg to about 350 mg. This dosage regimen can be adjusted to provide the best therapeutic effect.

The crystal form or pharmaceutical composition thereof disclosed in the present invention can be effectively used to prevent, treat, cure or alleviate FXR-mediated diseases in patients, and can particularly effectively treat 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, cirrhosis, hepatitis B, sebaceous gland disease, alcohol-induced cirrhosis, cystic fibrosis, biliary obstruction, cholelithiasis, liver fibrosis, dyslipidemia, atherosclerosis, type II diabetes, diabetic nephropathy, diabetic neuropathy, Diabetic retinopathy, peripheral arterial occlusive disease (PAOD), colitis, neonatal jaundice, nuclear jaundice, veno-occlusive disease, portal hypertension, metabolic syndrome, acute myocardial infarction, acute stroke, thrombosis, hypercholesterolemia, intestinal bacterial overgrowth, erectile dysfunction, gastrointestinal tumor diseases and liver tumor diseases, etc.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is an X-ray powder diffraction (XRPD) pattern of Form II of the compound represented by formula (I).

FIG2 is a differential scanning calorimetry (DSC) diagram of Form II of the compound represented by formula (I).

FIG3 is a thermogravimetric analysis (TGA) diagram of the crystalline form II of the compound represented by formula (I).

FIG4 is an X-ray powder diffraction (XRPD) pattern of Form III of the compound represented by formula (I).

FIG5 is a differential scanning calorimetry (DSC) diagram of Form III of the compound represented by formula (I).

FIG6 is a thermogravimetric analysis (TGA) diagram of Form III of the compound represented by formula (I).

FIG7 is an X-ray powder diffraction (XRPD) pattern of Form VII of the compound represented by formula (I).

FIG8 is a differential scanning calorimetry (DSC) diagram of Form VII of the compound represented by formula (I).

FIG9 is a thermogravimetric analysis (TGA) diagram of Form VII of the compound represented by formula (I).

FIG10 is an X-ray powder diffraction (XRPD) pattern of Form VIII of the compound represented by formula (I).

FIG11 is a differential scanning calorimetry (DSC) diagram of Form VIII of the compound represented by formula (I).

FIG12 is a thermogravimetric analysis (TGA) diagram of Form VIII of the compound represented by formula (I).

FIG13 is an X-ray powder diffraction (XRPD) pattern of Form IX of the compound represented by formula (I).

FIG14 is a differential scanning calorimetry (DSC) diagram of Form IX of the compound represented by formula (I).

FIG15 is an X-ray powder diffraction (XRPD) pattern of Form I of the compound represented by formula (I).

Detailed ways

The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples.

The X-ray powder diffraction analysis method used in the present invention is: Empyrean diffractometer, using Cu-Kα radiation (45KV, 40mA) to obtain X-ray powder diffraction patterns. 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 a range of 3°-40° with a step size of 0.0168°. Data Collector software is used to collect data, HighScore Plus software is used to process data, and Data Viewer software is used to read data.

The differential scanning calorimetry (DSC) analysis method used in the present invention is: 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. About 1-5 mg of sample is accurately weighed into a special aluminum crucible with a lid, and a linear heating device of 10°C/min is used to analyze the sample from room temperature to about 300°C. During use, the DSC chamber is purged with dry nitrogen.

Example

Unless otherwise stated, the compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (i.e., the compound of formula (I)) in the following examples is prepared by referring to the synthesis method of Example 3 in patent applications WO 2016127924A1 and CN 105884758A, and is identified by Empyrean X-ray powder diffraction (XRPD) analysis as Form I, having an X-ray powder diffraction (XRPD) pattern substantially as shown in Figure 15, as described in Example 6 of the present invention.

Example 1 Crystalline Form II of the Compound Represented by Formula (I)

1. Preparation of Form II

Method 1: Add the compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (900.0 mg) to methyl acetate (22.0 mL), heat under reflux until the solid is completely dissolved, then cool to room temperature, and then add n-heptane (22.0 mL), and solid precipitates. Filter by suction, and dry the filter cake under vacuum at room temperature to obtain a white solid (685.0 mg, yield 76.11%).

Method 2: Add compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (0.50 g) to methyl acetate (10.0 mL), heat under reflux with stirring until the solid is completely dissolved, keep warm for 10 minutes, stop heating, cool naturally to room temperature and stir for 6 hours, filter, and dry the filter cake in vacuum at room temperature for 6 hours to obtain a white solid (0.36 g, yield 72.0%).

2. Identification of Form II

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: Using Cu-Kα radiation, it has the following characteristic peaks expressed in degrees 2θ: 5.65°, 9.63°, 11.36°, 11.84°, 12.11°, 12.71°, 13.45°, 14.69°, 14.96°, 16.42°, 16.97°, 17.18°, 17.77°, 18.26°, 18.89°, 19.30°, 20.11°, 20.77°, 21.08°, 21.31°, 21.90°, 22.81°, 23.56°, 24.31°, 24.52°, 25.29° ,26.36°,26.67°,27.03°,27.69°,27.97°,28.33°,28.67°,29.11°,29.91°,30.29°,30.62°,31.54°,31.81°,32.35°,33.15°,33.79°,34.29°,35.10°,35.75°,36.64°,37.99°,38.82°,39.67°,40.78°,43.11°,43.90°,44.95°,45.90°,48.79°,51.65°,52.76° and 53.79°, with an error tolerance of ±0.2°.

(2) The product was analyzed and identified by TA Q2000 differential scanning calorimetry (DSC): the scanning speed was 10°C/min, and the resulting DSC curve was shown in FIG2 , which contained endothermic peaks at 125.84°C and 198.41°C, with an error tolerance of ±3°C.

(3) Thermogravimetric analysis (TGA) was performed using TA Q500: the heating rate was 10°C/min, and the resulting TGA curve is shown in Figure 3. The weight loss range was 7.23%, with an error tolerance of ±0.1%.

Example 2 Crystalline Form III of the Compound Represented by Formula (I)

1. Preparation of Form III

The compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (900.0 mg) was added to ethyl formate (20.0 mL), heated under reflux until the solid was completely dissolved, and then slowly cooled to room temperature, and solid precipitated. The filter cake was vacuum dried at room temperature to obtain a white solid (571.0 mg, yield 63.35%).

2. Identification of Form III

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis using Cu-Kα radiation, the following characteristic peaks are expressed in degrees 2θ: 5.70°, 9.01°, 9.59°, 10.95°, 11.43°, 11.95°, 12.15°, 13.55°, 14.18°, 14.82°, 15.09°, 15.56°, 16.50°, 17.17°, 17.74°, 17.88°, 18.37°, 19.03°, 19.40°, 20.13°, 20.92°, 21.12°, 21.48°, 21.81°, 22.90°, 23.51°, 24.38°,24.67°,25.19°,25.98°,26.21°,26.83°,27.21°,27.50°,28.01°,28.59°,28.92°,29.45°,29.84°,30.79°,31.37°,31.74°,32.47°,33.58°,34.08°,35.83°,36.47°,38.11°,38.93°,40.60°,42.59°,42.79°,43.63°,45.20°,46.52°,48.74°,51.65° and 53.46°, with an error tolerance of ±0.2°.

(2) The product was analyzed and identified by TA Q2000 differential scanning calorimetry (DSC): the scanning speed was 10°C/min, and the resulting DSC curve was shown in FIG5 , which contained endothermic peaks at 104.93°C and 199.52°C, with an error tolerance of ±3°C.

(3) Thermogravimetric analysis (TGA) was performed using TA Q500: the heating rate was 10°C/min, and the resulting TGA curve was shown in Figure 6. The weight loss range was 4.383%, with an error tolerance of ±0.1%.

Example 3 Crystalline Form VII of the Compound Represented by Formula (I)

1. Preparation of Form VII

Method 1: Add the compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (30.0 g) to dimethyl sulfoxide (150.0 mL), and the solid is completely dissolved. After stirring for 4 hours, water (450.0 mL) is added, and stirring is continued for 40 hours. Filter with suction, and the filter cake is vacuum dried at 60°C for 16 hours to obtain a white solid (29.51 g, yield 98.38%).

Method 2: Add the compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (1.0 g) to dimethyl sulfoxide (2.0 mL), the solid is completely dissolved, N,N-dimethylacetamide (2.0 mL) is added and heated to 60°C, kept warm for 2 hours and then slowly cooled to room temperature, water (12.0 mL) is added and stirred for 16 hours. Filter by suction, and dry the filter cake under vacuum at 60°C for 4 hours to obtain a white solid (0.93 g, yield 93.00%).

Method 3: Add the compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (52.0 mg) to N,N-dimethylacetamide (0.5 mL), heat to 90°C and then slowly cool to room temperature, add water (1.5 mL), and solid precipitates. Filter by suction, and dry the filter cake under vacuum at room temperature to obtain a white solid (42.1 mg, yield 80.96%).

Method 4: Add the compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (50.0 mg) to N,N-dimethylformamide (0.5 mL), heat to 60°C, keep warm for 2 hours, then slowly cool to room temperature, add water (1.5 mL) to crystallize, filter, and dry the filter cake in vacuo at room temperature to obtain a white solid (46.6 mg, yield 93.20%).

Method 5: Add 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (5.08 g) and isopropanol (84.0 mL) into a 100 mL two-necked bottle, heat to 79°C, wait until the solid is completely dissolved, add the above solution into 5-10°C water (198.0 mL), continue stirring for 1 hour after the addition, then stir at room temperature overnight, filter, and dry the filter cake under vacuum at room temperature for 6 hours to obtain an off-white solid (4.33 g, yield 85.23%).

2. Identification of Form VII

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-Kα radiation, with the following angles expressed in 2θ Characteristic peaks: 8.88°, 10.57°, 10.90°, 11.87°, 12.91°, 13.92°, 14.93°, 16.00°, 16.35°, 17.10°, 17.97°, 18.57°, 18.86°, 19.48°, 20.60°, 21.08°, 21.72°, 22.19°, 22.66°, 23.23°, 23.55°, 24.06°, 25.04°, 25.69°, 26.42°, 26.75°, 27.14°, 27.93°, 28.47°, 28.71°, 29.06°, 29.39°, 3 0.25°,30.64°,31.47°,31.82°,32.19°,32.84°,33.95°,34.51°,35.43°,35.83°,36.28°,36.61°,37.12°,37.89°,38.82°,39.64°,40.16°,40.68°,41.10°,41.53°,43.14°,44.15°,45.71°,46.33°,46.83°,47.35°,51.15°,52.63°,54.22°,55.16° and 57.64°, with an error tolerance of ±0.2°.

(2) The product was analyzed and identified by TA Q2000 differential scanning calorimetry (DSC): the scanning speed was 10°C/min, and the resulting DSC curve was shown in FIG8 , which contained endothermic peaks at 177.53°C and 199.42°C and an exothermic peak at 179.32°C±3°C, with an error tolerance of ±3°C.

(3) Thermogravimetric analysis (TGA) was performed using TA Q500: the heating rate was 10°C/min, and the resulting TGA curve was shown in Figure 9. The weight loss range was 0.3968%, with an error tolerance of ±0.1%.

Example 4 Crystalline Form VIII of the Compound Represented by Formula (I)

1. Preparation of Form VIII

The compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (250.0 mg) was added to butanone (5.0 mL) and heated to 60°C. The solid was completely dissolved. Then ethylene glycol monoethyl ether (5.0 mL) was added first, and then water (7.5 mL) was added. A small amount of solid precipitated. After keeping warm for 2 hours, the temperature was slowly cooled to room temperature. Filtered with suction, the filter cake was dried under vacuum at room temperature to obtain a white solid (199.3 mg, yield 79.72%).

2. Identification of Form VIII

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-Kα radiation, the following characteristic peaks are expressed in angles 2θ: 5.70°, 9.60°, 11.34°, 11.82°, 12.12°, 12.71°, 13.35°, 14.75°, 14.89°, 16.48°, 17.24° ,17.75°,18.08°,19.00°,19.24°,19.98°,20.92°,21.25°,21.87°,22.73°,23.65°,24.33°,24.60°,25.19°,25.58°,26.31°,26.54°,26.91°,27.55°,28.09° ,28.51°,28.79°,29.01°,29.30°,29.96°,30.85°,31.47°,31.80°,32.17°,32.81°,33.66°,34.15°,34.85°,35.63°,35.94°,36.54°,36.91°,38.00°,39.08° , 40.96°, 42.55°, 42.92°, 43.63°, 44.88°, 46.17°, 46.47°, 48.68°, 50.02°, 51.23°, 51.77°, 52.40°, 53.28°, 53.72°, 54.70°, 56.96° and 58.34°, with an error tolerance of ±0.2°.

(2) The product was analyzed and identified by TA Q2000 differential scanning calorimetry (DSC): the scanning speed was 10°C/min, and the resulting DSC curve was shown in FIG11 , which contained endothermic peaks at 116.13°C, 191.72°C, and 199.49°C, with an error tolerance of ±3°C.

(3) Thermogravimetric analysis (TGA) was performed using TA Q500: the heating rate was 10°C/min, and the resulting TGA curve was shown in Figure 12. The weight loss range was 2.081%, with an error tolerance of ±0.1%.

Example 5 Crystalline Form IX of the Compound Represented by Formula (I)

1. Preparation of Form IX

Compound 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy-10,11-dihydrobenzo[6,7]oxazo[3,2-b]pyridine-7-carboxylic acid (500.1 mg) and ethyl formate (12.5 mL) were added to a 50 mL two-necked bottle, heated under reflux with stirring, and the solid was completely dissolved and kept warm for 1 hour, then cooled to room temperature and stirred for 6 hours, filtered, and pumped to near dryness. The filter cake was vacuum dried at 60° C. for 6 hours to obtain a white solid (410.8 mg, yield 82.16%).

2. Identification of Form IX

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis using Cu-Kα radiation, the following characteristic peaks are expressed in angles 2θ: 5.30°, 10.33°, 10.59°, 11.12°, 11.95°, 12.96°, 13.16°, 13.31°, 13.48°, 15.92°, 16.45°, 17.18°, 19.52°, 19. .90°,20.23°,20.68°,21.17°,21.27°,21.82°,22.08°,22.20°,22.44°,23.39°,23.49°,24.02°,24.26°,24.89°,25.26°,25.90°,26.30°,26.46°,26.75°,27.16°,27.55°,28.72° ,29.34°,29.81°,30.22°,30.42°,31.24°,32.10°,32.68°,32.92°,33.25°,33.59°,34.05°,34.81°,35.20°,35.82°,36.13°,36.36°,37.59°,38.27°,39.05°,39.57°,40.21°,41 .78°, 42.15°, 42.63°, 43.21°, 43.94°, 44.92°, 45.94°, 46.83°, 47.44°, 49.08°, 49.65°, 50.40°, 50.71°, 52.27°, 53.05°, 53.89°, 54.66°, 55.83°, 57.68° and 59.39°, with an error tolerance of ±0.2°.

(2) The product was analyzed and identified by TA Q2000 differential scanning calorimetry (DSC): the scanning speed was 10°C/min, and the resulting DSC curve was shown in FIG14 , which contained endothermic peaks at 111.76°C, 193.61°C, and 199.64°C, with an error tolerance of ±3°C.

Example 6 Crystalline Form I of the Compound Represented by Formula (I)

1. Preparation of Form I

It was prepared by referring to the synthesis method of Example 3 in patent applications WO 2016127924A1 and CN 105884758A.

2. Identification of Form I

Identification by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-Kα radiation. The X-ray powder diffraction pattern of Form I of the compound represented by formula (I) prepared in this embodiment is shown in FIG15 .

Example 7 Pharmacokinetics of the Crystalline Form of the Present Invention

experimental method:

The crystal form of the compound represented by formula (I) of the present invention is filled into capsules for oral administration.

Six male Beagle dogs weighing 8-12 kg were taken, 3 in each group, and capsules containing the test sample were orally administered at a dose of 9 mg/kg. Blood was collected at time points of 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 h. A standard curve with an appropriate range was established based on the sample concentration. The concentration of the test sample in the plasma sample was determined in the MRM mode using the AB SCIEX API4000 LC-MS/MS, and quantitative analysis was performed. According to the drug concentration-time curve, the pharmacokinetic parameters were calculated using the non-compartmental model method using WinNonLin 6.3 software. The experimental results are shown in Table 1.

Table 1 Pharmacokinetic experimental data of the crystal form of the present invention

Experimental results:

As can be seen from Table 1, the crystal form VII of the present invention has a high exposure amount and a high blood drug concentration in beagle dogs, and has better pharmacokinetic properties than the prior art.

Example 8 Stability Test

(1) High temperature test: Take an appropriate amount of the test sample and put it into a flat weighing bottle, spread it into a thin layer ≤5 mm thick, and place it at 60℃±2℃ for 30 days. Take samples on the 0th, 5th, 10th and 30th days for testing according to the key stability inspection items, observe the color change of the sample, and test the sample purity by HPLC.

(2) High humidity test: Take an appropriate amount of the test sample and put it into a flat weighing bottle, spread it into a thin layer ≤5 mm thick, and place it under 25°C and RH90%±5% for 30 days. Take samples on the 0th, 5th, 10th and 30th days for testing according to the key stability inspection items, observe the color change of the sample, and test the sample purity by HPLC.

(3) Light test: Place the samples under the conditions of visible light illumination of 4500 lx ± 500 lx and ultraviolet light energy of not less than 0.7 W·h/ ^m2 for 30 days. Take samples at 0, 5, 10, 15 and 30 days for testing according to the key stability inspection items, observe the color changes of the samples, and test the purity of the samples by HPLC.

The experimental results show that the crystalline form VII of the present invention has good stability under various sample preparation conditions and is suitable for pharmaceutical use.

The above contents are only basic descriptions of the concept of the present invention, and any equivalent changes made according to the technical solution of the present invention shall fall within the protection scope of the present invention.

In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example" or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims (12)

1. A crystalline form of the compound represented by formula (I), wherein the crystalline form is crystalline form VII, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 13.92°±0.2°, 14.93°±0.2°, 19.48°±0.2°, 23.23°±0.2°, 23.55°±0.2°;
   
2. The crystalline form according to claim 1, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0.2°, 19.48°±0.2°, 20.60°±0.2°, 22.19°±0.2°, 22.66°±0.2°, 23.23°±0.2°, 23.55°±0.2°, 24.06°±0.2°, 26.42°±0.2°, 26.75°±0.2°.
3. The crystalline form according to claim 1 or 2, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0.2°, 17.97°±0.2°, 18.57°±0.2°, 18.86°±0.2°, 19.48°±0.2°, 20.60°±0.2°, 21.72°±0.2°, 22.19 °±0.2°,22.66°±0.2°,23.23°±0.2°,23.55°±0.2°,24.06°±0.2°,25.04°±0.2°,25.69°±0.2°,26.42°±0.2°,26.75°±0.2°,27.93°±0.2°,30.25°±0.2°,31.47°±0.2°,32.19°±0.2°,35.83°±0.2°,36.61°±0.2°.
4. The crystalline form according to any one of claims 1 to 3, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 10.57°±0.2°, 10.90°±0.2°, 11.87°±0.2°, 12.91°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0.2°, 16.35°±0.2°, 17.10°±0.2 °,17.97°±0.2°,18.57°±0.2°,18.86°±0.2°,19.48°±0.2°,20.60°±0.2°,21.08°±0.2°,21.72°±0.2°,22.19°±0.2°,22.66°±0.2°,23.23°±0.2°,23.55°±0.2°,24.06°±0.2°,25.04°±0.2°,25.69°±0.2°,26. 42°±0.2°,26.75°±0.2°,27.14°±0.2°,27.93°±0.2°,28.47°±0.2°,28.71°±0.2°,29.06°±0.2°,29.39°±0.2°,30.25°±0.2°,30.64°±0.2°,31.47°±0.2°,31.82°±0.2°,32.19°±0.2°,32.84°±0.2°,33.95°± 0.2°,34.51°±0.2°,35.43°±0.2°,35.83°±0.2°,36.28°±0.2°,36.61°±0.2°,37.12°±0.2°,37.89°±0.2°,38.82°±0.2°,40.68°±0.2°,41.10°±0.2°,41.53°±0.2°,43.14°±0.2°,47.35°±0.2°,51.15°±0.2°.
5. The crystalline form according to any one of claims 1 to 4, characterized in that the X-ray powder diffraction pattern of the crystalline form VII comprises the following diffraction peaks at 2θ angles: 8.88°±0.2°, 10.57°±0.2°, 10.90°±0.2°, 11.87°±0.2°, 12.91°±0.2°, 13.92°±0.2°, 14.93°±0.2°, 16.00°±0. 2°,16.35°±0.2°,17.10°±0.2°,17.97°±0.2°,18.57°±0.2°,18.86°±0.2°,19.48°±0.2°,20.60°±0.2°,21.08°±0.2°,21.72°±0.2°,22.19°±0.2°,22.66°±0.2°,23.23°±0.2°, 23.55°±0.2°,24.06°±0.2°,25.04°±0.2°,25.69°±0.2°,26.42°±0.2°,26.75°±0.2°,27.14°±0.2°,27.93°±0.2°,28.47°±0.2°,28.71°±0.2°,29.06°±0.2°,29.39°±0.2°,30. 25°±0.2°,30.64°±0.2°,31.47°±0.2°,31.82°±0.2°,32.19°±0.2°,32.84°±0.2°,33.95°±0.2°,34.51°±0.2°,35.43°±0.2°,35.83°±0.2°,36.28°±0.2°,36.61°±0.2°,37.12° ±0.2°,37.89°±0.2°,38.82°±0.2°,39.64°±0.2°,40.16°±0.2°,40.68°±0.2°,41.10°±0.2°,41.53°±0.2°,43.14°±0.2°,44.15°±0.2°,45.71°±0.2°,46.33°±0.2°,46.83°±0.2°,47.35°±0.2°,51.15°±0.2°,52.63°±0.2°,54.22°±0.2°,55.16°±0.2°,57.64°±0.2°.
6. The crystalline form according to any one of claims 1 to 5, characterized in that the crystalline form VII has an X-ray powder diffraction pattern substantially as shown in Figure 7.
7. The crystalline form according to any one of claims 1 to 6, characterized in that the differential scanning calorimetry diagram of the crystalline form VII comprises endothermic peaks at 177.53°C ± 3°C and 199.42°C ± 3°C and an exothermic peak at 179.32°C ± 3°C.
8. The crystalline form according to any one of claims 1 to 7, characterized in that the crystalline form VII has a differential scanning calorimetry diagram substantially as shown in Figure 8.
9. A pharmaceutical composition comprising the crystal form according to any one of claims 1 to 8, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant or a combination thereof.
10. Use of the crystalline form according to any one of claims 1 to 8 or the pharmaceutical composition according to claim 9 in the preparation of a drug for preventing, treating or alleviating a disease mediated by FXR in a patient.
11. The use according to claim 10, wherein the disease mediated by FXR is cardiovascular and cerebrovascular disease, a disease related to dyslipidemia, metabolic syndrome, a hyperproliferative disease, fibrosis, an inflammatory disease or a liver and gallbladder-related disease.
12. The use according to claim 11, wherein the cardiovascular and cerebrovascular diseases are atherosclerosis, acute myocardial infarction, veno-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 levels of fatty acids or glycerol 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;

    The hyperproliferative diseases are hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma, breast cancer, pancreatic cancer, Bart's esophagus cancer and other forms of gastrointestinal or liver neoplastic diseases;

    The fibrosis, inflammatory disease or hepatobiliary-related disease is 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 damage, gallstones, cirrhosis, hepatitis B, sebaceous gland disease, alcohol-induced cirrhosis, biliary obstruction, cholelithiasis, colitis, neonatal jaundice, nuclear jaundice or intestinal bacterial overgrowth.