CRYSTALLINE FORM OF NITROGEN-CONTAINING TRICYCLIC COMPOUND AND USE THEREOF

Abstract

A crystalline form A of a nitrogen-containing tricyclic compound shown in formula (I), a pharmaceutical composition containing the crystalline form A, and the use of the crystalline form A or the pharmaceutical composition in the preparation of a medication for preventing, treating or relieving an FXR-mediated disease of a patient.

Claims

1. A crystal form of a compound having Formula (I), wherein the crystal form is crystal form A, ##STR00003## wherein the crystal form A exhibits the following characteristic X-ray powder diffraction peaks expressed as 2θ at 14.07°±0.2°, 21.00°±0.2°, 22.59°±0.2°, 22.86°±0.2°, 26.30°±0.20.

2. The crystal form according to claim 1, wherein the crystal form A exhibits the following characteristic X-ray powder diffraction peaks expressed as 2θ at 11.11°±0.2°, 14.07°±0.2°, 20.37°±0.2°, 21.00°±0.2°, 22.59°±0.2°, 22.86°±0.2°, 23.32°±0.2°, 24.85°±0.2°, 25.57°±0.2°, 26.11°±0.2°, 26.30°±0.2°, 28.92°±0.2°, 38.74°±0.2°.

3. The crystal form according to claim 1, wherein the crystal form A exhibits the following characteristic X-ray powder diffraction peaks expressed as 2θ at 7.22°±0.2°, 8.65°±0.2°, 11.11°±0.2°, 12.61°±0.2°, 12.92°±0.2°, 13.86°±0.2°, 14.07°±0.2°, 14.37°±0.2°, 15.94°±0.2°, 16.68°±0.2°, 17.29°±0.2°, 18.95°±0.2°, 19.90°±0.2°, 20.37°±0.2°, 20.89°±0.2°, 21.00°±0.2°, 21.60°±0.2°, 22.24°±0.2°, 22.59°±0.2°, 22.86°±0.2°, 23.32°±0.2°, 24.28°±0.2°, 24.85°±0.2°, 25.29°±0.2°, 25.57°±0.2°, 26.11°±0.2°, 26.30°±0.2°, 26.77°±0.2°, 27.62°±0.2°, 27.94°±0.2°, 28.15°±0.2°, 28.92°±0.2°, 30.61°±0.2°, 31.13°±0.2°, 31.41°±0.2°, 32.14°±0.2°, 33.65°±0.2°, 34.39°±0.2°, 34.73°±0.2°, 35.54°±0.2°, 36.38°±0.2°, 36.80°±0.2°, 37.76°±0.2°, 38.24°±0.2°, 38.74°±0.2°, 39.48°±0.2°, 40.08°±0.2°, 40.59°±0.2°, 42.34°±0.2°, 43.30°±0.2°, 43.99°±0.2°, 44.53°±0.2°, 45.29°±0.2°, 46.10°±0.2°, 47.83°±0.2°, 48.64°±0.2°, 49.91°±0.2°, 51.75°±0.2°, 54.43°±0.2°, 58.46°±0.2°.

4. The crystal form according to claim 1, wherein the crystal form A has an X-ray powder diffraction pattern substantially as shown in FIG. 1.

5. A crystal form of a compound having Formula (I), wherein the crystal form is crystal form A, ##STR00004## wherein the crystal form A has a differential scanning calorimetry thermogram comprising an endothermic peak at 196.35° C.±3° C.

6. The crystal form according to claim 5, wherein the crystal form A has a differential scanning calorimetry thermogram substantially as shown in FIG. 2.

7. A pharmaceutical composition comprising the crystal form of claim 1, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant or a combination thereof.

8. A method for preventing, treating or lessening a disease mediated by FXR in a patient, comprising administering a pharmaceutically acceptable effective dose of the crystal form of claim 1.

9. The method according to claim 8, wherein the disease mediated by FXR is cardiovascular and cerebrovascular disease, a disease related to dyslipidemia, metabolic syndrome, hyperproliferative disease, fibrosis, inflammatory disease or a disease related to liver and gallbladder.

10. The method according to claim 9, wherein the cardiovascular and cerebrovascular disease is atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke or thrombosis; wherein the metabolic syndrome is insulin resistance, hyperglycemia, hyperinsulinemia, elevated levels of fatty acid 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 comorbidities of diabetes and abnormally high body mass index; wherein the hyperproliferative disease is hepatocellular carcinoma, colonic adenocarcinoma, polyposis, colonic adenocarcinoma, breast cancer, membrane adenocarcinoma, Barrett's esophagus cancer, or other forms of gastrointestinal or liver neoplastic diseases; wherein the fibrosis, inflammatory disease or disease related to liver and gallbladder is non-alcoholic fatty liver, non-alcoholic steatohepatitis, cholestasis, liver fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial intrahepatic cholestasis, cystic fibrosis, drug-induced bile duct damage, gallstones, liver cirrhosis, hepatitis B, sebaceous gland disease, alcohol-induced liver cirrhosis, bile duct obstruction, gallstone disease, colitis, neonatal Jaundice, nuclear jaundice, or overgrowth of intestinal bacteria.

11. A method for preventing, treating or lessening a disease mediated by FXR in a patient, comprising administering a pharmaceutically acceptable effective dose of the pharmaceutical composition of claim 7.

12. The method according to claim 11, wherein the disease mediated by FXR is cardiovascular and cerebrovascular disease, a disease related to dyslipidemia, metabolic syndrome, hyperproliferative disease, fibrosis, inflammatory disease or a disease related to liver and gallbladder.

13. The method according to claim 12, wherein the cardiovascular and cerebrovascular disease is atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke or thrombosis; wherein the metabolic syndrome is insulin resistance, hyperglycemia, hyperinsulinemia, elevated levels of fatty acid 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 comorbidities of diabetes and abnormally high body mass index; wherein the hyperproliferative disease is hepatocellular carcinoma, colonic adenocarcinoma, polyposis, colonic adenocarcinoma, breast cancer, membrane adenocarcinoma, Barrett's esophagus cancer, or other forms of gastrointestinal or liver neoplastic diseases; wherein the fibrosis, inflammatory disease or disease related to liver and gallbladder is non-alcoholic fatty liver, non-alcoholic steatohepatitis, cholestasis, liver fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial intrahepatic cholestasis, cystic fibrosis, drug-induced bile duct damage, gallstones, liver cirrhosis, hepatitis B, sebaceous gland disease, alcohol-induced liver cirrhosis, bile duct obstruction, gallstone disease, colitis, neonatal Jaundice, nuclear jaundice, or overgrowth of intestinal bacteria.

14. A pharmaceutical composition comprising the crystal form of claim 5, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant or a combination thereof.

15. A method for preventing, treating or lessening a disease mediated by FXR in a patient, comprising administering a pharmaceutically acceptable effective dose of the crystal form of claim 5.

16. The method according to claim 15, wherein the disease mediated by FXR is cardiovascular and cerebrovascular disease, a disease related to dyslipidemia, metabolic syndrome, hyperproliferative disease, fibrosis, inflammatory disease or a disease related to liver and gallbladder.

17. The method according to claim 16, wherein the cardiovascular and cerebrovascular disease is atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke or thrombosis; wherein the metabolic syndrome is insulin resistance, hyperglycemia, hyperinsulinemia, elevated levels of fatty acid 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 comorbidities of diabetes and abnormally high body mass index; wherein the hyperproliferative disease is hepatocellular carcinoma, colonic adenocarcinoma, polyposis, colonic adenocarcinoma, breast cancer, membrane adenocarcinoma, Barrett's esophagus cancer, or other forms of gastrointestinal or liver neoplastic diseases; wherein the fibrosis, inflammatory disease or disease related to liver and gallbladder is non-alcoholic fatty liver, non-alcoholic steatohepatitis, cholestasis, liver fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial intrahepatic cholestasis, cystic fibrosis, drug-induced bile duct damage, gallstones, liver cirrhosis, hepatitis B, sebaceous gland disease, alcohol-induced liver cirrhosis, bile duct obstruction, gallstone disease, colitis, neonatal Jaundice, nuclear jaundice, or overgrowth of intestinal bacteria.

18. A method for preventing, treating or lessening a disease mediated by FXR in a patient, comprising administering a pharmaceutically acceptable effective dose of the pharmaceutical composition of claim 14.

19. The method according to claim 18, wherein the disease mediated by FXR is cardiovascular and cerebrovascular disease, a disease related to dyslipidemia, metabolic syndrome, hyperproliferative disease, fibrosis, inflammatory disease or a disease related to liver and gallbladder.

20. The method according to claim 19, wherein the cardiovascular and cerebrovascular disease is atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke or thrombosis; wherein the metabolic syndrome is insulin resistance, hyperglycemia, hyperinsulinemia, elevated levels of fatty acid 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 comorbidities of diabetes and abnormally high body mass index; wherein the hyperproliferative disease is hepatocellular carcinoma, colonic adenocarcinoma, polyposis, colonic adenocarcinoma, breast cancer, membrane adenocarcinoma, Barrett's esophagus cancer, or other forms of gastrointestinal or liver neoplastic diseases; wherein the fibrosis, inflammatory disease or disease related to liver and gallbladder is non-alcoholic fatty liver, non-alcoholic steatohepatitis, cholestasis, liver fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial intrahepatic cholestasis, cystic fibrosis, drug-induced bile duct damage, gallstones, liver cirrhosis, hepatitis B, sebaceous gland disease, alcohol-induced liver cirrhosis, bile duct obstruction, gallstone disease, colitis, neonatal Jaundice, nuclear jaundice, or overgrowth of intestinal bacteria.

Description

DESCRIPTION OF THE DRAWINGS

[0064] FIG. 1 is an X-ray powder diffraction (XRPD) pattern of the crystal form A of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

[0065] FIG. 2 is a differential scanning calorimetry (DSC) thermogram of the crystal form A of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

[0066] FIG. 3 is a thermogravimetric analysis (TGA) thermogram of the crystal form A of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

[0067] FIG. 4 shows the changes of X-ray powder diffraction pattern of the crystal form A of the compound having Formula (I) during the stability experiment under high temperature condition according to the method (1) of Example 3 of the present invention.

[0068] FIG. 5 shows the changes of X-ray powder diffraction pattern of the crystal form A of the compound having Formula (I) during the stability experiment under high humidity condition according to the method (2) of Example 3 of the present invention.

[0069] FIG. 6 shows the changes of X-ray powder diffraction pattern of the crystal form A of the compound having Formula (I) during the stability experiment under light condition according to the method (3) of Example 3 of the present invention.

[0070] FIG. 7 is a Dynamic Vapor Sorption (DVS) graph of the crystal form A of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

EMBODIMENTS

[0071] The invention will now be further described by way of examples without limiting the invention to the scope of the examples.

[0072] The X-ray powder diffraction analysis method used in the present invention was an Empyrean diffractometer, and an X-ray powder diffraction pattern was obtained using Cu-Kα radiation (45 KV, 40 mA). The powdery sample was prepared as a thin layer on a monocrystalline silicon sample rack and placed on a rotating sample stage, analyzed at a rate of 0.0167 steps in the range of 3°-60°. Data Collector software was used to collect data, HighScore Plus software was used to process data, and Data Viewer software was used to read data.

[0073] The differential scanning calorimetry (DSC) analysis method used in the present invention was performing a differential scanning calorimetry analysis using a TA Q2000 module with a thermal analysis controller. Data were collected and analyzed using TA Instruments Thermal Solutions software. Approximately 1-5 mg of the sample was accurately weighed into a specially crafted aluminum crucible with a lid and analyzed from room temperature to about 300° C. using a linear heating device at 10° C./min. During use, the DSC chamber was purged with dry nitrogen.

[0074] The thermogravimetric analysis (TGA) analysis method used in the present invention is performing a thermogravimetric analysis using a TA Q500 module with a thermal analysis controller. Data were collected and analyzed using TA Instruments Thermal Solutions software. Approximately 10-30 mg of the sample was placed into a platinum crucible and analyzed from room temperature to about 300° C. using a linear heating device at 10° C./min. During use, the DSC chamber was purged with dry nitrogen.

Specific Embodiments

[0075] The specific synthetic method of compound having Formula (I) named 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepino[3,2-b]pyridine-11,1′-cyclopropane]-7-carboxylic acid refers to example 9 in patent application CN 107686486.

EXAMPLES

Example 1 a Crystal Form A of the Compound Having Formula (I)

1. Preparation of Crystal Form A

[0076] Compound

2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro [benzo[6,7]oxepino[3,2-b]pyridine-11,1′-cyclopropane]-7-carboxylic acid (201.5 mg) was added into isopropanol (4.0 mL) at room temperature, then the mixture was refluxed with stirring to dissolve the material, then heating was stopped. The mixture was naturally cooled to room temperature and stirred for crystallization for 12 hours. The resulting mixture was filtered by suction, and the filter cake was dried over in vacuo at 50° C. overnight to give a white solid (120.3 mg, 59.7%).

2. Identify of Crystal Form A

[0077] (1) The crystal form A was identified by Empyrean X-ray powder diffraction (XRPD) analysis using Cu-Kα radiation, and the crystal form A exhibited the following characteristic peaks at angles of 2θ: 7.22°, 8.65°, 11.110, 12.610, 12.92°, 13.86°, 14.07°, 14.37°, 15.94°, 16.68°, 17.29°, 18.95°, 19.90°, 20.37°, 20.89°, 21.00°, 21.60°, 22.24°, 22.59°, 22.86°, 23.32°, 24.28°, 24.85°, 25.29°, 25.57°, 26.11°, 26.30°, 26.77°, 27.62°, 27.94°, 28.15°, 28.92°, 30.61°, 31.13°, 31.41°, 32.14°, 33.65°, 34.39°, 34.73°, 35.54°, 36.38°, 36.80°, 37.76°, 38.24°, 38.74°, 39.48°, 40.08°, 40.59°, 42.34°, 43.30°, 43.99°, 44.53°, 45.29°, 46.10°, 47.83°, 48.64°, 49.91°, 51.75°, 54.430 and 58.46°, there was an error tolerance of ±0.2°. The X-ray powder diffraction of the crystal form A prepared by the method of the example was substantially as shown in FIG. 1.

[0078] (2) The crystal form A was analyzed and identified by TA Q2000 Differential Scanning Calorimetry (DSC): the scanning speed was 10° C./min, and the crystal form A had an endothermic peak of 196.35° C., and there was an error tolerance of ±3° C. The differential scanning calorimetry thermogram of the crystal form A prepared by the method of the example was substantially as shown in FIG. 2.

[0079] (3) The crystal form A was analyzed and identified by Thermogravimetric Analysis (TGA) using TA Q500: the heating rate was 10° C./min, the weight loss was 0.121%, and there was an error tolerance of ±0.1%. The thermogravimetric Analysis graph of the crystal form A prepared by the method of the example was substantially as shown in FIG. 3.

Example 2 the Pharmacokinetic Test of the Crystal Form A of the Present Invention

[0080] The crystal form A of compound having Formula (I) named 2-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-10H-spiro[benzo[6,7]oxepino[3,2-b]pyridine-11,1′-cyclopropane]-7-carboxylic acid was filled into capsules, which was for oral administration.

[0081] 8-12 kg Male Beagle dogs were given 5 mg/kg test sample orally, with 3 animals in each group. Blood samples were taken at time points of 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 12 and 24 h. Standard curve was plotted based on concentrations of the samples in a suitable range, the concentration of the test sample in the plasma sample was measured and quantified by AB SCIEX API4000 LC-MS/MS at MRM mode. Pharmacokinetic parameters were calculated according to drug concentration-time curve using a noncompartmental model by WinNonLin 6.3 software. Results were as shown in Table 1.

TABLE-US-00002 TABLE 1 Pharmacokinetic data of the crystal form A of the present invention Test sample dosage (mg/kg) AUC.sub.last (h*ng/ml) C.sub.max (ng/ml) T.sub.max (h) Crystal form A 5 237 86 0.667

Conclusion

[0082] It can be seen from Table 1 that the crystal form A of the present invention has a relatively large exposure in Beagle dogs and has good pharmacokinetic properties.

Example 3 the Stability Test of the Crystal Form A of the Present Invention

[0083] (1) High-temperature test: A batch of the test sample was taken into a flat weighing bottle, divided into ≤5 mm thick thin layer. The sample was placed at 60° C. for 30 days. The sample was then sampled at day 5, day 10 and day 30, and the change of the sample's color was observed. The purity of the sample was detected by HPLC, and the structure of the sample was analyzed by X-ray powder diffraction, the change of X-ray powder diffraction in high temperature test was substantial as shown in FIG. 4.

[0084] (2) High-humidity test: A batch of the test sample was taken into a flat weighing bottle, divided into ≤5 mm thick thin layer. The sample was placed at 25° C., RH 90%±5% for 30 days. The sample was then sampled at day 5, day 10 and day 30, and the change of the sample's color was observed. The purity of the sample was detected by HPLC, and the structure of the sample was analyzed by X-ray powder diffraction, the change of X-ray powder diffraction in high humidity test was substantial as shown in FIG. 5.

[0085] (3) Light test: A batch of the test sample was taken into a flat weighing bottle, divided into ≤5 mm thick thin layer. The sample was placed in an opened light box (with UV) at the illuminance 4500±500lx, UV light ≥0.7 w/m.sup.2 for 30 days. The sample was then sampled at day 5, day 13 and day 30, and the change of the sample's color was observed. The purity of the sample was detected by HPLC, and the structure of the sample was analyzed by X-ray powder diffraction, the change of X-ray powder diffraction in light test was substantial as shown in FIG. 6.

[0086] The changes of appearance and chemical purity of the test samples in the stability test were as shown in Table 2.

TABLE-US-00003 TABLE 2 The stability test of the crystal form A of the present invention Condition High-temperature High-humidity Light 0 5 10 30 5 10 30 5 13 30 Project days days days days days days days days days days Appearance White White White White White White White White White White powder powder powder powder powder powder powder powder powder powder Purity (%) 99.44 99.33 99.35 99.45 99.55 99.39 99.42 99.53 99.31 99.43

Conclusion

[0087] Under the conditions of high-temperature, high-humidity and light, the appearance, chemical purity and crystal form of the crystal form A of the present invention have no obvious changes, the stability effect is good, and it is suitable for pharmaceutical use.

Example 4 the Hygroscopicity Test of the Crystal Form A of the Present Invention

[0088] An appropriate amount of the test sample was taken and the hygroscopicity of the test sample was tested by dynamic moisture adsorption device. Results were as shown in FIG. 7. It's demonstrated from the test results that the crystal form A of the present invention has a weight gain of less than 0.1% after equilibrium under the condition of a relative humidity of 80%. According to the criterion for the hygroscopic weight gain, it has no or almost none hygroscopicity. That is, the crystal form A of the invention is not easily deliquescent under the influence of high humidity.

[0089] The foregoing description is merely a basic illustration of the present invention and any equivalent transformation made in accordance with the technical solution of the present invention is intended to be within the scope of the present invention.

[0090] Reference throughout this specification to “an embodiment,” “some embodiments,” “one embodiment”, “another example,” “an example,” “a specific example,” or “some examples,” means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present disclosure. Thus, the appearances of the above terms throughout this specification are not necessarily referring to the same embodiment or example of the present disclosure. Furthermore, the particular features, structures, materials, or characteristics may be combined in any suitable manner in one or more embodiments or examples. In addition, those skilled in the art can integrate and combine different embodiments, examples or the features of them as long as they are not contradictory to one another.

[0091] Although explanatory embodiments have been shown and described, it would be appreciated by those skilled in the art that the above embodiments cannot be construed to limit the present disclosure, and changes, alternatives, and modifications can be made in the embodiments without departing from spirit, principles and scope of the present disclosure.