AMORPHOUS FORM OF NITROGEN-CONTAINING TRICYCLIC COMPOUND AND USE THEREOF

Abstract

An amorphous form of a nitrogen-containing tricyclic compound and a use thereof, a pharmaceutical composition containing the compound in the amorphous form, and the use of the compound in the amorphous form or the pharmaceutical composition in the preparation of a drug for preventing, treating or alleviating FXR-mediated diseases in a patient.

Claims

1. An amorphous form of a compound having Formula (I), ##STR00003## wherein the amorphous form has an X-ray powder diffraction pattern substantially as shown in FIG. 1.

2. An amorphous form of a compound having Formula (I), ##STR00004## wherein the amorphous form has a glass transition temperature of 92.26° C. ±3° C.

3. The amorphous form according to claim 2, wherein the amorphous form has a differential scanning calorimetry thermogram substantially as shown in FIG. 2.

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

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

6. The method according to claim 5, 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.

7. The method according to claim 6, 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 a comorbidity 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 disease; 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.

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 pharmaceutical composition of claim 4.

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 a comorbidity 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 disease; 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 pharmaceutical composition comprising the amorphous form of claim 2, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant or a combination thereof.

12. A method for preventing, treating or lessening a disease mediated by FXR in a patient, comprising administering a pharmaceutically acceptable effective dose of the amorphous form of claim 2.

13. The method according to claim 12, 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.

14. The method according to claim 13, 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 a comorbidity 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 disease; 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.

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 pharmaceutical composition of claim 11.

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 a comorbidity 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 disease; 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

[0057] FIG. 1 is an X-ray powder diffraction (XRPD) pattern of the amorphous form of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

[0058] FIG. 2 is a differential scanning calorimetry (DSC) thermogram of the amorphous form of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

[0059] FIG. 3 is a thermogravimetric analysis (TGA) thermogram of the amorphous form of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

[0060] FIG. 4 shows the changes of X-ray powder diffraction pattern of the amorphous form 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.

[0061] FIG. 5 shows the changes of X-ray powder diffraction pattern of the amorphous form 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.

[0062] FIG. 6 shows the changes of X-ray powder diffraction pattern of the amorphous form 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.

[0063] FIG. 7 is a dynamic vapor sorption (DVS) graph of the amorphous form of the compound having Formula (I) prepared according to the method of Example 1 of the present invention.

DESCRIPTION OF THE PREFERRED EMBODIMENT

[0064] The invention will now be further described by way of examples without limiting the invention to the scope of the described examples.

[0065] 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, High Score Plus software was used to process data, and Data Viewer software was used to read data.

[0066] 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.

[0067] 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

[0068] 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.

EXAMPLE S

Example 1 An Amorphous Form of the Compound Having Formula (I)

1. Preparation of Amorphous Form

[0069] Compound 2-((5-cyclopropyl-3-(2, 6-dichlorophenyl)i soxazol-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 a 100 mL single-necked flask at room temperature, then a solution of sodium hydroxide (133.8 mg, 3.34 mmol) in water (30.0 mL) was added. The mixture was heated to 65° C. until the solid was dissolved completely, then cooled to 40° C.; and then the self-made diluted hydrochloric acid (4.74 mmol, 15.0 mL) was added dropwise. After addition, the pH of the mixture was 2-3, then the mixture was stopped heating, and naturally cooled to room temperature and stirred for crystallization for 3 hours. The resulting mixture was filtered by suction, and the filter cake was washed with water (3.0 mL×3), dried over in vacuo at 60° C. for 6 hours to give a white solid (1.41 g, 92.2%).

2. Identification of Amorphous Form

[0070] (1) The amorphous form was identified by Empyrean X-ray powder diffraction (XRPD) analysis using Cu-Ka radiation, and the X-ray powder diffraction was substantially as shown in FIG. 1.

[0071] (2) The amorphous form was analyzed and identified by TA Q2000 Differential Scanning calorimetry (DSC): the scanning speed was 10° C./min, and the amorphous form had a glass transition temperature of 92.26° C., and there was an error tolerance of ±3° C. The differential scanning calorimetry thermogram of the amorphous form prepared by the method of the example was substantially as shown in FIG. 2.

[0072] (3) The amorphous form was analyzed and identified by Thermogravimetric Analysis (TGA) using TA Q500: the heating rate was 10° C./min, the weight loss was 0.409%, and there was an error tolerance of ±0.1%. The thermogravimetric analysis graph of the amorphous form prepared by the method of the example was substantially as shown in FIG. 3.

Example 2 The Pharmacokinetic Test of the Amorphous Form of the Present Invention

[0073] The amorphous form of compound having Formula (I) named 2-((5-cyclopropyl-3-(2,6-di chl orophenyl)i soxazol-4-yl)methoxy)-10H-spiro[b enzo[6,7]oxepino[3,2-b]pyridine-11,1′-cyclopropane]-7-carboxylic acid was filled into capsules, which was for oral administration.

[0074] 8-12 kg Male Beagle dogs were given 5 mg/kg test sample orally, with 3 animals in each group. Blood was 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 amorphous form of the present invention dosage AUC.sub.last C.sub.max T.sub.max Test sample (mg/kg) (h*ng/ml) (ng/ml) (h) The amorphous form 5 765 268 2

[0075] Conclusion:

[0076] It can be seen from Table 1 that the amorphous form of the present invention has a relatively large exposure in Beagle dogs and has good pharmacokinetic properties.

Example 3 The Stability Test of the Amorphous Form of the Present Invention

[0077] (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 taken on days 5, 10, and 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 substantially shown in FIG. 4.

[0078] (2) High humidity test: A batch of the test sample was taken into the flat weighing bottle, divided into <5 mm thick thin layer. The sample was placed at 25° C. and RH for 90% ±5% for 30 days. The sample was then taken on days 5, 10, and 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 substantially shown in FIG. 5.

[0079] (3) Light test: A batch of the test sample was taken into the 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±5001x, UV light ≥0.7w/m2 for 30 days. The sample was then taken on days 5, 13, and 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 substantially shown in FIG. 6.

[0080] 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 amorphous form of the present invention Condition High temperature High humidity Light Project 0 Day 5 Days 10 Days 30 Days 5 Days 10 Days 30 Days 5 Days 13 days 30 days Appearance White White White White White White White White White White powder powder powder powder powder powder powder powder powder powder Purity (%) 99.66 99.65 99.48 99.60 99.64 99.61 99.63 99.62 99.63 99.57

[0081] Conclusion:

[0082] Under the conditions of high temperature, high humidity and light, the appearance, chemical purity and crystal form of the amorphous form 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 Amorphous Form of the Present Invention

[0083] 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 table 7. It's demonstrated from the test results that the amorphous form of the present invention has a weight gain of less than 2% after equilibrium under the condition of a relative humidity of 80%. According to the criterion for the hygroscopic weight gain, it is slightly hygroscopic. That is, the amorphous form of the invention is not easily deliquescent under the influence of high humidity.

[0084] 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.

[0085] 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.

[0086] 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.