Crystalline forms of a bromodomain and extraterminal protein inhibitor drug, processes for preparation thereof, and use thereof

Abstract

The present disclosure relates to novel crystalline forms of a bromodomain protein inhibitor 2-[4-(2-hydroxyethoxy)-3,5-dimethylphenyl]-5,7-dimethoxyquinazolin-4(3H)-one and processes for preparation and use thereof, relating to pharmaceutical industry. Form CS2, Form CS8, Form CS13, Form CS20, Form CS1, Form CS7, Form CS9, Form CS11 and Form CS4 of the present disclosure can be used for preparing drugs treating cardiovascular, cholesterol or lipid-related disorders. ##STR00001##

Claims

1. A crystalline form CS1 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 6.10.2, 12.30.2, 26.10.2 and 26.80.2 using CuK radiation.

2. The crystalline form CS1 according to claim 1, wherein the X-ray powder diffraction pattern shows one or more characteristic peaks at 2theta values of 16.40.2, 18.50.2, 23.20.2, 13.00.2, 14.10.2, 17.10.2 and 24.50.2 using CuK radiation.

3. A crystalline form CS2 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 11.50.2, 6.60.2 and 8.80.2 using CuK radiation.

4. The crystalline form CS2 according to claim 3, wherein the X-ray powder diffraction pattern shows one or two characteristic peaks at 2theta values of 5.10.2 and 15.30.2 using CuK radiation.

5. A crystalline form CS8 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 23.90.2, 13.50.2, 7.80.2, 22.50.2 and 11.40.2 using CuK radiation.

6. The crystalline form CS8 according to claim 5, wherein the X-ray powder diffraction pattern shows one or more characteristic peaks at 2theta values of 25.90.2, 13.10.2, 28.10.2 and 20.20.2 using CuK radiation.

7. A crystalline form CS13 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 5.10.2, 12.50.2 and 17.10.2 using CuK radiation.

8. The crystalline form CS13 according to claim 7, wherein the X-ray powder diffraction pattern shows one or more characteristic peaks at 2theta values of 6.40.2, 8.50.2, 25.70.2, 7.80.2 and 16.00.2 using CuK radiation.

9. A crystalline form CS20 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 8.40.2, 18.90.2 and 13.50.2 using CuK radiation.

10. The crystalline form CS20 according to claim 9, wherein the X-ray powder diffraction pattern shows one or more characteristic peaks at 2theta values of 11.30.2, 9.40.2, 5.60.2, 26.30.2, 20.10.2, 20.60.2 and 24.40.2 using CuK radiation.

11. A crystalline form CS7 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 5.90.2, 6.70.2, 10.70.2 and 12.50.2 using CuK radiation.

12. The crystalline form CS7 according to claim 11, wherein the X-ray powder diffraction pattern shows one or two or three characteristic peaks at 2theta values of 8.40.2, 16.90.2, 13.30.2, 16.00.2, 25.10.2, 15.00.2 and 21.80.2 using CuK radiation.

13. A crystalline form CS9 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 7.30.2, 9.90.2 and 17.00.2 using CuK radiation.

14. The crystalline form CS9 according to claim 13, wherein the X-ray powder diffraction pattern shows one or more characteristic peaks at 2theta values of 13.40.2, 3.90.2, 12.80.2, 12.10.2, 24.90.2, 22.50.2 and 24.20.2 using CuK radiation.

15. A crystalline form CS11 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 7.80.2, 8.80.2, 9.70.2 and 13.60.2 using CuK radiation.

16. The crystalline form CS11 according to claim 15, wherein the X-ray powder diffraction pattern shows one or more characteristic peaks at 2theta values of 4.40.2, 16.90.2, 21.60.2, 13.00.2 and 15.30.2 using CuK radiation.

17. A crystalline form CS4 of apabetalone, wherein the X-ray powder diffraction pattern shows characteristic peaks at 2theta values of 9.10.2, 14.50.2, 23.50.2 and 24.20.2 using CuK radiation.

18. The crystalline form CS4 according to claim 17, wherein the X-ray powder diffraction pattern shows one or two or three characteristic peaks at 2theta values of 10.30.2, 25.00.2, 26.30.2, 10.80.2, 11.60.2 and 19.50.2 using CuK radiation.

19. A method for treating cardiovascular, cholesterol or lipid-related disorders, comprising administering to a patient in need thereof a therapeutically effective amount of the Form CS1 according to claim 1.

20. A method for treating cardiovascular, cholesterol or lipid-related disorders, comprising administering to a patient in need thereof a therapeutically effective amount of the Form CS7 according to claim 11.

21. A method for treating cardiovascular, cholesterol or lipid-related disorders, comprising administering to a patient in need thereof a therapeutically effective amount of the Form CS4 according to claim 17.

22. A method for treating cardiovascular, cholesterol or lipid-related disorders, comprising administering to a patient in need thereof a therapeutically effective amount of the Form CS9 according to claim 13.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1A shows an XRPD pattern of Form CS2 according to example 1 of the present disclosure.

(2) FIG. 1B shows a DSC curve of Form CS2 according to example 1 of the present disclosure.

(3) FIG. 1C shows a TGA curve of Form CS2 according to example 1 of the present disclosure.

(4) FIG. 2A shows an XRPD pattern of Form CS8 according to example 2 of the present disclosure.

(5) FIG. 2B shows a DSC curve of Form CS8 according to example 2 of the present disclosure.

(6) FIG. 2C shows a TGA curve of Form CS8 according to example 2 of the present disclosure.

(7) FIG. 2D shows an XRPD pattern overlay of Form CS8 before and after being stored under 25 C./60% RH for two weeks (from top to bottom: XRPD pattern before storage, XRPD pattern after being stored under 25 C./60% RH for two weeks).

(8) FIG. 2E shows a DVS plot of Form CS8 according to example 2 of the present disclosure.

(9) FIG. 2F shows an XRPD pattern overlay of Form CS8 according to example 2 of the present disclosure before and after DVS test.

(10) FIG. 3A shows an XRPD pattern of Form CS13 according to example 3 of the present disclosure.

(11) FIG. 3B shows a DSC curve of Form CS13 according to example 3 of the present disclosure.

(12) FIG. 3C shows a TGA curve of Form CS13 according to example 3 of the present disclosure.

(13) FIG. 4A shows an XRPD pattern of Form CS20 according to example 4 of the present disclosure.

(14) FIG. 4B shows a DSC curve of Form CS20 according to example 4 of the present disclosure.

(15) FIG. 4C shows a TGA curve of Form CS20 according to example 4 of the present disclosure.

(16) FIG. 4D shows a .sup.1H NMR spectrum of Form CS20 according to example 4 of the present disclosure.

(17) FIG. 4E shows an XRPD pattern overlay of Form CS20 before and after being stored under 25 C./60% RH and 40 C./75% RH for two weeks (from top to bottom: XRPD pattern before storage, XRPD pattern after being stored under 25 C./60% RH for two weeks, XRPD pattern after being stored under 40 C./75% RH for two weeks).

(18) FIG. 5A shows an XRPD pattern of Form CS1 according to example 5 method 1 of the present disclosure.

(19) FIG. 5B shows a DSC curve of Form CS1 according to example 5 method 1 of the present disclosure.

(20) FIG. 5C shows a TGA curve of Form CS1 according to example 5 method 1 of the present disclosure.

(21) FIG. 5D shows an XRPD pattern of Form CS1 according to example 5 method 2 of the present disclosure.

(22) FIG. 5E shows an XRPD pattern overlay of Form CS1 before and after being stored under 25 C./60% RH and 40 C./75% RH for 10 months (from top to bottom: XRPD pattern before storage, XRPD pattern after being stored under 25 C./60% RH for 10 months, XRPD pattern after being stored under 40 C./75% RH for 10 months).

(23) FIG. 5F shows a DVS plot of Form CS1 according to example 5 of the present disclosure.

(24) FIG. 5G shows an XRPD pattern overlay of Form CS1 according to example 5 of the present disclosure before and after DVS test.

(25) FIG. 5H shows an XRPD pattern overlay of Form CS1 according to example 5 of the present disclosure before and after grinding.

(26) FIG. 5I shows a dissolution rate profile of Form CS1 according to example 5 of the present disclosure in formulations.

(27) FIG. 6A shows an XRPD pattern of Form CS7 according to example 6 of the present disclosure.

(28) FIG. 6B shows a DSC curve of Form CS7 according to example 6 of the present disclosure.

(29) FIG. 6C shows a TGA curve of Form CS7 according to example 6 of the present disclosure

(30) FIG. 6D shows an XRPD pattern overlay of Form CS7 before and after being stored under 25 C./60% RH and 40 C./75% RH for 4 weeks and 80 C. for one day (from top to bottom: XRPD pattern before storage, XRPD pattern after being stored under 25 C./60% RH for 4 weeks, XRPD pattern after being stored under 40 C./75% RH for 4 weeks, XRPD pattern after being stored under 80 C. for one day).

(31) FIG. 6E shows an XRPD pattern overlay of Form CS7 according to example 6 of the present disclosure before and after DVS test.

(32) FIG. 7A shows an XRPD pattern of Form CS9 according to example 7 of the present disclosure.

(33) FIG. 7B shows a DSC curve of Form CS9 according to example 7 of the present disclosure.

(34) FIG. 7C shows a TGA curve of Form CS9 according to example 7 of the present disclosure.

(35) FIG. 7D shows an XRPD pattern overlay of Form CS9 before and after being stored under 25 C./60% RH and 40 C./75% RH for 10 months (from top to bottom: XRPD pattern before storage, XRPD pattern after being stored under 25 C./60% RH for 10 months, XRPD pattern after being stored under 40 C./75% RH for 10 months).

(36) FIG. 7E shows a DVS plot of Form CS9 according to example 7 of the present disclosure.

(37) FIG. 7F shows an XRPD pattern overlay of Form CS9 according to example 7 of the present disclosure before and after DVS test

(38) FIG. 8A shows an XRPD pattern of Form CS11 according to example 8 method 1 of the present disclosure

(39) FIG. 8B shows a DSC curve of Form CS11 according to example 8 method 1 of the present disclosure

(40) FIG. 8C shows a TGA curve of Form CS11 according to example 8 method 1 of the present disclosure

(41) FIG. 8D shows an XRPD pattern of Form CS11 according to example 8 method 2 of the present disclosure

(42) FIG. 8E shows an XRPD pattern overlay of Form CS11 before and after being stored under 25 C./60% RH and 40 C./75% RH for 6 weeks (from top to bottom: XRPD pattern before storage, XRPD pattern after being stored under 25 C./60% RH for 6 weeks, XRPD pattern after being stored under 40 C./75% RH for 6 weeks).

(43) FIG. 9A shows an XRPD pattern of Form CS4 according to example 9 of the present disclosure

(44) FIG. 9B shows a DSC curve of Form CS4 according to example 9 of the present disclosure

(45) FIG. 9C shows a TGA curve of Form CS4 according to example 9 of the present disclosure

(46) FIG. 9D shows an XRPD pattern overlay of Form CS4 before and after being stored under 25 C./60% RH and 40 C./75% RH for 10 months (from top to bottom: XRPD pattern before storage, XRPD pattern after being stored under 25 C./60% RH for 10 months, XRPD pattern after being stored under 40 C./75% RH for 10 months).

(47) FIG. 9E shows a DVS plot of Form CS4 according to example 9 of the present disclosure

(48) FIG. 9F shows an XRPD pattern overlay of Form CS4 according to example 9 of the present disclosure before and after DVS test.

(49) FIG. 9G shows an XRPD pattern overlay of Form CS4 according to example 9 of the present disclosure before and after grinding.

DETAILED DESCRIPTION

(50) The present disclosure is further illustrated by the following examples which describe the preparation and use of the crystalline forms of the present disclosure in detail. It is obvious to those skilled in the art that many changes in the materials and methods can be accomplished without departing from the scope of the present disclosure.

(51) The abbreviations used in the present disclosure are explained as follows:

(52) XRPD: X-ray Powder Diffraction

(53) HPLC: High Performance Liquid Chromatography

(54) DVS: Dynamic Vapor Sorption

(55) TGA: Thermal Gravimetric Analysis

(56) .sup.1H NMR: Proton Nuclear Magnetic Resonance

(57) RH: Relative Humidity

(58) Instruments and Methods Used for Data Collection:

(59) X-ray powder diffraction patterns in the present disclosure were acquired by a Panalytical Empyrean X-ray powder diffractometer. The parameters of the X-ray powder diffraction method of the present disclosure were as follows: X-ray Reflection: Cu, K K1 (): 1.5400598; K2 (): 1.544426 K2/K1 intensity ratio: 0.50 Voltage: 45 (kV) Current: 40 (mA) Scan range: from 3.0 degree to 40.0 degree

(60) Differential scanning calorimetry (DSC) data in the present disclosure were acquired by a TA Instruments Q200 MDSC. Instrument control software is Thermal Advantage, and analysis software is Universal Analysis.

(61) Heating rate: 10 C./min

(62) Purge gas: nitrogen

(63) Thermal gravimetric analysis (TGA) data in the present disclosure were acquired by a TA Instruments Q500 TGA. Instrument control software is Thermal Advantage, and analysis software is Universal Analysis. Heating rate: 10 C./min Purge gas: nitrogen

(64) Proton nuclear magnetic resonance spectrum data (.sup.1H NMR) were collected from a Bruker Avance II DMX 400M HZ NMR spectrometer. 1-5 mg of sample was weighed, and dissolved in 0.5 mL of deuterated dimethyl sulfoxide to obtain a solution with a concentration of 2-10 mg/mL.

(65) Dynamic Vapor Sorption (DVS) was measured via a SMS (Surface Measurement Systems Ltd.) intrinsic DVS instrument. Its control software is DVSIntrinsic control software, and its analysis software is DVSIntrinsic Analysis software. Typical Parameters for DVS test are as follows: Temperature: 25 C. Gas and flow rate: N.sub.2, 200 mL/min dm/dt: 0.002%/min RH range: 0% RH to 95% RH

(66) High Performance Liquid Chromatography (HPLC) data in the present disclosure were collected from an Agilent 1260 with Diode Array Detector (DAD). The HPLC method parameters for purity test in the present disclosure are as follows: 1. Column: Waters XBridge C18 1504.6 mm, 5 m 2. Mobile Phase: A: 0.1% TFA in H.sub.2O B: 0.1% TFA in Acetonitrile

(67) Gradient:

(68) TABLE-US-00001 Time (min) % B 0.0 20 5.0 30 6.0 80 8.0 80 8.1 20 10.0 20 3. Flow rate: 1.6 mL/min 4. Injection Volume: 5 L 5. Detection wavelength: 260 nm 6. Column Temperature: 40 C. 7. Diluent: Acetonitrile/H.sub.2O (v/v, 1/1)

(69) The dissolution testing in the present disclosure was performed on an Agilent 708-DS.

(70) Unless otherwise specified, the following examples were conducted at room temperature.

(71) Raw materials of apabetalone used in the following examples were prepared by known methods disclosed in CN101641339B.

Example 1

(72) Preparation of Form CS2

(73) Certain amount of apabetalone was weighed and dissolved in corresponding solvent of Table 1.1 at T.sub.1. The solution was filtered and cooled to T.sub.2 slowly or rapidly. When precipitation occurred, solids was obtained after centrifugation and drying.

(74) TABLE-US-00002 TABLE 1.1 Sample Weight Volume Cooling T.sub.1 T.sub.2 No. (mg) Solvent (v/v) (mL) rate ( C.) ( C.) 1-a 19.7 Methanol 1.0 Rapid 50 20 1-b 10.2 Methanol/methyl 1.0 Slow 50 5 isobutyl ketone (1:1) 1-c 10.2 Methanol/methyl 1.0 Rapid 50 20 isobutyl ketone (1:1) 1-d 20.2 Methanol/toluene 1.0 Slow 50 5 (1:1) 1-e 20.2 Methanol/toluene 1.0 Rapid 50 20 (1:1) 1-f 11.3 Methanol/2-methyl 1.0 Slow 50 5 tetrahydrofuran (1:1) 1-g 11.3 Methanol/2-methyl 1.0 Rapid 50 20 tetrahydrofuran (1:1) 1-h 10.7 Tetrahydrofuran/ 3.0 Slow 50 5 methyl tert-butyl ether (2:1) 1-i 19.9 Methanol 1.0 Rapid 100 20

(75) Sample 1-a to 1-i were confirmed to be Form CS2 by XRPD. Sample 1-i was selected for characterization. The XRPD pattern is substantially as depicted in FIG. 1A, and the XRPD data are listed in Table 1.2. The DSC curve of Form CS2 is substantially as depicted in FIG. 1B, which shows the first endothermic peak at around 60 C., the second endothermic peak at around 113 C., the first exothermic peak at around 126 C. and the third endothermic peak at around 232 C. The TGA curve of Form CS2 is substantially as depicted in FIG. 1C, which shows about 3.4% weight loss when heated to 100 C.

(76) TABLE-US-00003 TABLE 1.2 2 Theta d spacing Intensity % 5.08 17.39 30.31 6.57 13.46 100.00 8.85 10.00 34.56 11.54 7.67 41.68 13.31 6.65 8.72 15.28 5.80 14.13 20.22 4.39 7.21 23.10 3.85 4.08 25.35 3.51 2.44
Solubility Study of Form CS2

(77) The prepared apabetalone Form CS2 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 1.3.

(78) TABLE-US-00004 TABLE 1.3 Solubility (mg/mL) Time (h) SGF FeSSIF 1 0.49 0.15 24 0.61 0.24

(79) The above results show that Form CS2 of apabetalone has good solubility in SGF and FeSSIF, especially in SGF, the solubility at 24 h is as high as 0.61 mg/mL. Polymorph with high solubility is conducive to increasing the blood concentration of drugs in human body and improves the bioavailability of drugs, which is of great significance for drug research.

Example 2

(80) Preparation of Form CS8

(81) Certain amount of apabetalone was weighed and dissolved in corresponding solvent of Table 2.1 at T.sub.1. The solution was filtered and cooled to T.sub.2 slowly or rapidly. When precipitation occurred, solids was obtained after centrifugation and drying.

(82) TABLE-US-00005 TABLE 2.1 Sample Weight Volume Cooling T.sub.1 T.sub.2 No. (mg) Solvent (v/v) (mL) rate ( C.) ( C.) 2-a 9.8 Dichloromethane 1.5 Slow 50 5 2-b 9.8 Dichloromethane 1.5 Rapid 50 20 2-c 20.8 Dichloromethane/ 1.0 Slow 50 5 isopropanol (4:1) 2-d 20.8 Dichloromethane/ 1.0 Rapid 50 20 isopropanol (4:1)

(83) Sample 2-a to 2-d were confirmed to be Form CS8 by XRPD. Sample 2-a was selected for characterization. The XRPD pattern is substantially as depicted in FIG. 2A, and the XRPD data are listed in Table 2.2. The DSC curve of Form CS8 is substantially as depicted in FIG. 2B, which shows the first exothermic peak at around 177 C., the first endothermic peak at around 211 C., the second exothermic peak at around 217 C. and the second endothermic peak at around 230 C. The TGA curve of Form CS8 is substantially as depicted in FIG. 2C, which shows about 0.4% weight loss when heated to 120 C.

(84) TABLE-US-00006 TABLE 2.2 2 Theta d spacing Intensity % 4.23 20.89 3.93 7.83 11.30 60.72 9.74 9.08 6.55 10.58 8.36 5.86 11.36 7.79 16.82 13.11 6.75 16.65 13.55 6.54 40.35 18.00 4.93 10.44 19.29 4.60 9.65 20.17 4.40 14.20 22.47 3.96 25.75 23.91 3.72 100.00 24.22 3.67 32.56 24.87 3.58 11.65 25.88 3.44 14.42 26.18 3.40 17.03 28.16 3.17 14.65 31.72 2.82 2.11
Stability Study of Form CS8
Stability Study of Apabetalone Form CS8

(85) Form CS8 was placed in a constant temperature and humidity chamber at 25 C./60% RH for 2 weeks in open dish. Crystalline form of the sample were tested by XRPD and impurity of the sample were checked. The results are shown in FIG. 2D (From top to bottom: XRPD pattern of Form CS8 before and after being stored under 25 C./60% RH for 2 weeks.)

(86) No obvious form change and purity decrease was observed for Form CS8 after being stored under 25 C./60% RH for 2 weeks. The results show that Form CS8 has good stability.

(87) Solubility Study of Form CS8

(88) The prepared Form CS8 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 2.3.

(89) TABLE-US-00007 TABLE 2.3 Solubility (mg/mL) Time (h) SGF FeSSIF 1 0.43 0.28

(90) The above results show that Form CS8 of apabetalone has good solubility in SGF and FeSSIF. Polymorph with high solubility is beneficial to increase the blood concentration of drugs in human body and improve the bioavailability of drugs, which is of great significance for drug research.

(91) Hygroscopicity of Form CS8

(92) Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS8 in the present disclosure with about 10 mg of sample. The result is listed in Table 2.4. The DVS plot of Form CS8 is substantially as depicted in FIG. 2E.

(93) TABLE-US-00008 TABLE 2.4 Weight Gain under 80% Form Relative Humidity Form CS8 0.34%

(94) Description and definition of hygroscopicity (Chinese Pharmacopoeia 2015 edition appendix 9103 drug hygroscopic test guidelines, test at 25 C.+/1 C., 80% RH.). deliquescent: Sufficient water is absorbed to form a liquid; very hygroscopic: Increase in mass is equal to or greater than 15 percent; hygroscopic: Increase in mass is less than 15 percent and equal to or greater than 2 percent; slightly hygroscopic: Increase in mass is less than 2 percent and equal to or greater than 0.2 percent. non hygroscopic or almost non hygroscopic: Increase in mass is less than 0.2%.

(95) The results indicates that the weight gain of Form CS8 under 80% RH is 0.34%. According to the definition of hygroscopicity, Form CS8 is slightly hygroscopic. The XRPD pattern of Form CS8 after DVS test is shown in FIG. 2F. No form change was observed for Form CS8 after DVS test, which indicates that Form CS8 is stable under the influence of humidity.

(96) Form CS8 of the present disclosure shows low hygroscopicity and can avoid the problems such as crystal instability in the process of drug preparation and/or storage, as well as the unprocessability of the preparation caused by external factors such as environmental moisture, which is conducive to the accurate quantitative preparation and later transportation and storage.

Example 3

(97) Preparation of Form CS13

(98) Certain amount of apabetalone was weighed and dissolved in corresponding solvent shown in Table 3.1. The solution was filtered and evaporated slowly at room temperature with or without addition of polymer to obtain solid.

(99) TABLE-US-00009 TABLE 3.1 Whether to add Sample Weight Solvent Volume polymer T No. (mg) (v/v) (mL) (Y/N) ( C.) 3-a 10.8 Tetrahydrofuran/ 1.3 N 25 water (4:1) 3-b 10.5 Acetone/ 3.3 N 25 water (9:1) 3-c 10.8 Tetrahydrofuran/ 1.3 Y 25 water (4:1) 3-d 10.5 Acetone/ 3.3 Y 25 water (9:1)

(100) Said polymer is a mixture of equal masses of polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate and hydroxyethyl cellulose.

(101) Sample 3-a to 3-d were confirmed to be Form CS13 by XRPD. Sample 3-d was selected for characterization. The XRPD pattern is substantially as depicted in FIG. 3A, and the XRPD data are listed in Table 3.2. The DSC curve of Form CS13 is substantially as depicted in FIG. 3B, which shows the first endothermic peak at around 70 C., the second endothermic peak at around 86 C., the third endothermic peak at around 170 C., the fourth endothermic peak at around 205 C., the first exothermic peak at around 207 C. and the fifth endothermic peak at around 230 C. The TGA curve of Form CS13 is substantially as depicted in FIG. 3C, which shows about 11.9% weight loss when heated to 100 C.

(102) TABLE-US-00010 TABLE 3.2 2 Theta d spacing Intensity % 5.09 17.35 48.06 6.38 13.85 22.94 7.76 11.39 4.83 8.54 10.35 12.25 10.17 8.70 0.71 11.48 7.71 3.89 12.54 7.06 100.00 13.29 6.66 3.59 15.30 5.79 1.21 15.56 5.69 2.14 16.01 5.54 4.65 17.13 5.18 47.37 19.15 4.64 0.27 20.41 4.35 1.35 20.84 4.26 4.59 22.11 4.02 0.59 23.43 3.80 3.37 25.67 3.47 8.18 26.75 3.33 1.24 27.06 3.29 3.65 28.72 3.11 1.18 29.56 3.02 1.42 30.89 2.90 0.43 32.26 2.77 1.36 34.67 2.59 0.58 37.28 2.41 2.20 38.97 2.31 0.89
Solubility of Form CS13

(103) The prepared Form CS13 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, 4 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 3.3.

(104) TABLE-US-00011 TABLE 3.3 Solubility Time (mg/mL) (h) SGF FeSSIF 1 0.35 0.13 4 0.31 0.14 24 0.33 0.14

(105) The above results show that Form CS13 of apabetalone has good solubility in SGF and FeSSIF.

Example 4

(106) Preparation of Form CS20

(107) Certain amount of apabetalone was weighed and dissolved in corresponding solvent of Table 4.1. The solution was filtered and evaporated slowly at room temperature to obtain a solid.

(108) TABLE-US-00012 TABLE 4.1 Sample Weight Solvent Volume T No. (mg) (v/v) (mL) ( C.) 4-a 10.5 Acetonitrile/acetic 2.0 25 acid (9:1) 4-b 10.3 Ethyl acetate/acetic 1.7 25 acid (4:1)

(109) Sample 4-a to 4-b were confirmed to be Form CS20 of apabetalone by XRPD. Sample 4-a was selected for characterization. The XRPD pattern is substantially as depicted in FIG. 4A, and the XRPD data are listed in Table 4.2. The DSC curve of Form CS20 is substantially as depicted in FIG. 4B, which shows the first endothermic peak at around 123 C., the second endothermic peak at around 185 C., the third endothermic peak at around 201 C., the fourth endothermic peak at around 207 C. and the fifth endothermic peak at around 230 C. The TGA curve of Form CS13 is substantially as depicted in FIG. 4C, which shows about 14.9% weight loss when heated to 160 C. The .sup.1H NMR spectrum of Form CS20 is substantially as depicted in FIG. 4D, and the corresponding data are: .sup.1H NMR (400 MHz, DMSO) 11.83 (s, 1H), 7.89 (s, 2H), 6.74 (d, J=2.3 Hz, 1H), 6.52 (d, J=2.3 Hz, 1H), 4.91 (s, 1H), 3.89 (s, 3H), 3.87-3.81 (m, 5H), 3.72 (s, 2H), 2.31 (s, 6H), 1.90 (s, 3H).

(110) TABLE-US-00013 TABLE 4.2 2 Theta d spacing Intensity % 5.59 15.81 21.68 7.20 12.28 4.52 8.39 10.53 100.00 9.37 9.44 22.64 11.26 7.86 25.84 11.67 7.58 8.95 13.52 6.55 83.84 13.88 6.38 7.82 14.48 6.12 11.79 14.92 5.94 3.42 16.92 5.24 11.24 18.88 4.70 30.97 19.57 4.54 1.85 20.15 4.41 12.22 20.56 4.32 12.06 22.30 3.99 5.29 22.76 3.91 10.73 24.10 3.69 2.41 24.38 3.65 4.37 25.26 3.53 1.52 25.93 3.44 3.39 26.33 3.38 13.70 26.78 3.33 3.08 28.11 3.17 2.02 28.81 3.10 3.75 34.30 2.61 1.52 35.68 2.52 2.00 36.85 2.44 2.14
Stability Study of Form CS20

(111) Two samples of apabetalone Form CS20 were placed in constant temperature and humidity chambers at 25 C./60% RH and 40 C./75% RH for 2 weeks in open dish. Crystalline form of the sample were tested by XRPD and impurity of the sample were checked. The XRPD pattern overlay is substantially as depicted in FIG. 4E (from top to bottom: XRPD pattern of Form CS20 before and after being stored under 25 C./60% RH and 40 C./75% RH for 2 weeks). No form change and obvious purity decrease was observed for Form CS20 after being stored at 25 C./60% RH and 40 C./75% RH for 2 weeks. It can be seen that Form CS20 has good stability.

(112) Solubility Study of Form CS20

(113) The prepared apabetalone Form CS20 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, 4 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 4.3.

(114) TABLE-US-00014 TABLE 4.3 Solubility Time (mg/mL) (h) SGF FeSSIF 1 0.44 0.19 4 0.41 0.16 24 0.51 0.22

(115) The above results show that Form CS20 of apabetalone of the present disclosure has good solubility in SGF and FeSSIF. Polymorph with high solubility is beneficial to increase the blood concentration of drugs in human body and improve the bioavailability of drugs, which is of great significance for drug research.

Example 5

(116) Preparation of Form CS1 (Method 1)

(117) The process of preparing Form CS1 of apabetalone comprises the following steps:

(118) Dissolving step: About 10 mg of apabetalone solid was dissolve in corresponding solvent of Table 5.1, and a clear solution was obtained by filtering.

(119) Precipitation step: The prepared solution was added to corresponding anti-solvent of Table 5.1 or corresponding anti-solvent was added to the prepared solution with stirring until a lot of precipitation was observed. The solid was collected by centrifugation and drying. The obtained solid was Form CS1 of apabetalone.

(120) Furthermore, the reaction conditions, composition and amount of solvents and anti-solvents of the preparing process for said Form CS1 of apabetalone are listed in Table 5.1.

(121) TABLE-US-00015 TABLE 5.1 Sample Weight Volume Anti- Volume No. (mg) Solvent (mL) solvent (mL) Method 5-a 10.1 Tetra- 2.125 n- 3.0 Anti-solvent hydrofuran Heptane addition 5-b 10.1 Tetra- 2.125 n- 3.0 Reverse hydrofuran Heptane anti-solvent addition 5-c 10.1 Tetra- 2.125 Methyl 3.0 Anti-solvent hydrofuran tertiary addition butyl ether 5-d 10.1 Tetra- 2.125 Methyl 3.0 Reverse hydrofuran tertiary anti-solvent butyl addition ether 5-e 10.1 Tetra- 2.125 Toluene 3.0 Reverse hydrofuran anti-solvent addition 5-f 10.1 Chloroform 1.375 Methyl 2.0 Reverse tertiary anti-solvent butyl addition ether 5-g 10.1 Chloroform 1.375 Toluene 3.0 Anti-solvent addition 5-h 9.9 Dimethyl 1.0 Water 3.0 Reverse sulfoxide anti-solvent addition 5-i 10.1 Dimethyl- 0.25 Aceto- 3.0 Reverse acetamide nitrile anti-solvent addition

(122) Sample 5-a to 5-i were confirmed to be Form CS1 of apabetalone by XRPD. Sample 5-a was selected for characterization. The XRPD pattern is substantially as depicted in FIG. 5A, and the XRPD data are listed in Table 5.2. The DSC curve of Form CS1 is substantially as depicted in FIG. 5B, which shows the first endothermic peak at around 207 C., the first exothermic peak at around 211 C. and the second endothermic peak at around 231 C. The TGA curve of Form CS1 is substantially as depicted in FIG. 5C, which shows about 0.7% weight loss when heated to 200 C.

(123) TABLE-US-00016 TABLE 5.2 2 Theta d spacing Intensity % 4.38 20.19 1.10 6.11 14.47 100.00 10.19 8.68 2.41 10.70 8.27 1.40 12.11 7.31 21.04 12.27 7.21 62.62 12.99 6.82 5.91 14.12 6.27 3.01 16.37 5.41 4.60 17.09 5.19 3.31 17.63 5.03 1.77 18.18 4.88 1.63 18.48 4.80 7.06 20.12 4.41 2.93 20.49 4.33 4.18 21.55 4.12 1.12 21.93 4.05 2.34 23.16 3.84 5.71 23.45 3.79 4.16 24.45 3.64 2.64 25.02 3.56 1.26 26.11 3.41 8.67 26.84 3.32 7.12 28.00 3.19 0.96 28.60 3.12 0.96 30.04 2.98 0.86 31.04 2.88 1.25 35.60 2.52 0.25 36.80 2.44 0.50 38.30 2.35 0.89
Preparation of Form CS1 (Method 2)

(124) The process of preparing Form CS1 comprises the following steps:

(125) Dissolving step: Apabetalone solid was dissolved in corresponding solvent of Table 5.3 at 50 C. to obtain a clear solution.

(126) Precipitation step: The solution was cooled to 20-5 C. rapidly or slowly until solid precipitated. The solid was collected by centrifugation and drying. The obtained solid was Form CS1 of apabetalone.

(127) Said reaction conditions, solvent composition and solvent amount of the preparation method of apabetalone Form CS1 are shown in Table 5.3. Sample 5-j to 5-n were confirmed to be Form CS1 of apabetalone by XRPD. The XRPD pattern of sample 5-j is substantially as depicted in FIG. 5D, and the XRPD data are listed in Table 5.4.

(128) TABLE-US-00017 TABLE 5.3 Sample Weight Solvent Volume T No. (mg) (v/v) (mL) Method ( C.) 5-j 19.9 Tetrahydrofuran 1.0 Slow 5 cooling 5-k 19.9 Tetrahydrofuran 1.0 Rapid 20 cooling 5-l 10.8 Acetone 3.0 Rapid 20 cooling 5-m 10.2 Ethyl acetate/ 3.0 Rapid 20 acetone (1:1) cooling 5-n 10.7 Acetonitrile/ 3.0 Rapid 20 N,N-dimethyl cooling formamide (9:1)

(129) TABLE-US-00018 TABLE 5.4 2 Theta d spacing Intensity % 3.57 24.78 1.09 6.10 14.48 100.00 10.26 8.62 0.50 12.28 7.21 65.97 13.01 6.81 2.85 14.15 6.26 2.43 16.36 5.42 3.05 17.13 5.18 2.82 18.47 4.80 6.67 19.24 4.61 0.64 20.14 4.41 0.92 20.51 4.33 1.86 22.01 4.04 1.19 23.16 3.84 7.43 24.45 3.64 2.81 25.01 3.56 1.21 26.15 3.41 10.57 26.86 3.32 8.29 28.76 3.10 0.82 30.08 2.97 0.92 31.07 2.88 1.86 34.62 2.59 0.49 36.83 2.44 0.62 38.30 2.35 1.75
Long-Term Stability Study of Form CS1

(130) Two samples of apabetalone Form CS1 were placed in constant temperature and humidity chambers at 25 C./60% RH and 40 C./75% RH for 10 months in open dishes. The samples were characterized by XRPD and chemical impurity. The results are substantially as depicted in FIG. 5E (from top to bottom: XRPD pattern of Form CS1 before and after being stored under 25 C./60% RH and 40 C./75% RH for 10 months) and Table 5.5.

(131) No form change and obvious purity decrease was observed for Form CS1 after being stored at 25 C./60% RH and 40 C./75% RH for 10 months. It can be seen that Form CS1 has good stability and high purity.

(132) TABLE-US-00019 TABLE 5.5 Condition 1 week 2 weeks 5 weeks 10 months 25 C./60% RH 99.27 99.23 99.25 99.24 40 C./75% RH 99.20 99.20 99.25 99.21
Solubility Study of Form CS1

(133) The prepared solid of apabetalone Form CS1 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, 4 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 5.6.

(134) TABLE-US-00020 TABLE 5.6 Solubility Time (mg/mL) (h) SGF FeSSIF 1 0.31 0.14 4 0.29 0.11 24 0.33 0.14

(135) The above results show that Form CS1 of apabetalone has good solubility in SGF and FeSSIF.

(136) Hygroscopicity Study of Form CS1

(137) Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS1 of the present disclosure with about 10 mg of sample. The result is listed in Table 5.7. The DVS plot of Form CS1 is substantially as depicted in FIG. 5F.

(138) TABLE-US-00021 TABLE 5.7 Weight Gain under 80% Form Relative Humidity Form CS1 0.13%

(139) The results showed that weight gain of Form CS1 under 80% RH is 0.13%. According to the hygroscopicity criteria, Form CS1 is almost non hygroscopic. The XRPD pattern of Form CS1 after DVS test is shown in FIG. 5G. No form change was observed for Form CS1 before and after DVS test, which indicates that Form CS1 is stable under the influence of humidity.

(140) Form CS1 of the present disclosure shows low hygroscopicity and can avoid the problems such as crystal instability in the process of drug preparation and/or storage, as well as the unprocessability of the preparation caused by external factors such as environmental moisture, which is conducive to the accurate quantitative preparation and later transportation and storage.

(141) Mechanical Stability of Form CS1

(142) Certain amount of Form CS1 was placed in a mortar and ground manually for 5 minutes. XRPD of the solid obtained was tested. The results are shown in FIG. 5H.

(143) The results showed that no form change and obvious crystallity decrease was observed for Form CS1 of apabetalone under certain mechanical stress, and it can still maintain stable physical and chemical properties, which is suitable for drug preparation and storage. Grinding of API is usually needed in the process of formulation, and good grinding stability will reduce the risk of crystallinity decrease and transformation of solid form of API.

(144) Formulation Study of Form CS1

(145) Certain amount of the API, microcrystalline cellulose, croscarmellose sodium according to Table 5.8 and 2 mg of magnesium stearate were weighted and blended for 2 minutes. Flakes were prepared using a manual tablet press at 5 KN pressure with a 20 mm round tooling. The crushed flakes were manually sieved through 20 mesh sieve. 2 mg of magnesium stearate was added, and then the obtained powder was blended for 1 minute. The mixture was encapsulated into a 1# capsule shell, then the capsule was packed in 35 cc HDPE (high density polyethylene) bottle (one capsule per bottle) containing 1 g of desiccant. The bottle was then sealed by sealing machine. The composition of the formulation (per 200 mg) is shown in table 5.8 below. Form CS1 is stable in the preparations after testing.

(146) TABLE-US-00022 TABLE 5.8 Quantity Component (mg/capsule) API 50 Microcrystalline cellulose 136 Croscarmellose sodium 10 Magnesium stearate 4
Dissolution test was performed on the obtained capsule. The conditions are as follows:
Medium: HCl (0.1 mol/L)
Method: Paddle
Volume: 900 mL
Speed: 75 rpm
Temperature: 37 C.

(147) Dissolution results of Form CS1 are presented in FIG. 5I, which indicates that Form CS1 possesses favorable dissolution. The released amount can reach more than 90% at 20 minutes. In drug product development, rapid dissolution rate can speed up the dissolution of the drug in body. By adjusting the excipients, it is possible to control the rapid action of drugs in specific parts and get a short onset of action of the drugs.

Example 6

(148) Preparation of Form CS7

(149) The process of preparing Form CS7 of apabetalone comprises the following steps:

(150) Dissolving step: 5.3 mg of apabetalone solid was dissolved in 1 mL of chloroform, and a clear solution was obtained by filtering.

(151) Precipitation step: The solution was added to a 3-mL glass vial. The vial was put into a 20-mL glass vial containing 5 mL of methyl isobutyl ketone for liquid vapor diffusion. Then the 20-mL vial was sealed and left at room temperature until solid precipitated. The solid was collected by centrifugation and drying to obtain Form CS7 of apabetalone. The XRPD pattern of Form CS7 is substantially as depicted in FIG. 6A, and the XRPD data are listed in Table 6.1.

(152) The DSC curve shows the first endothermic peak at around 231 C., which is substantially as depicted in FIG. 6B. The TGA curve shows about 0.4% weight loss when heated to 160 C., which is substantially as depicted in FIG. 6C.

(153) TABLE-US-00023 TABLE 6.1 2 Theta d spacing Intensity % 4.27 20.68 3.98 5.87 15.06 55.68 6.69 13.21 100 8.43 10.50 77.7 10.70 8.27 80.13 12.08 7.32 9.24 12.46 7.10 53.68 13.25 6.68 49.94 14.99 5.91 9.73 15.96 5.55 34.89 16.94 5.23 53.34 19.16 4.63 3.48 20.32 4.37 6.07 21.76 4.08 15.6 23.67 3.76 4.6 24.47 3.64 9.66 25.09 3.55 15.93 26.01 3.43 3.31 27.42 3.25 2.99 30.79 2.90 3.25 32.57 2.75 1.7 36.50 2.46 1.75 37.45 2.40 1.96
Stability Study of Form CS7

(154) Three samples of apabetalone Form CS7 were placed in constant temperature and humidity chambers at 25 C./60% RH and 40 C./75% RH for 4 weeks and 80 C. for 1 day in open dishes. Crystalline form of the sample were tested by XRPD and impurity of the sample were checked. The XRPD pattern overlay is substantially as depicted in FIG. 6D (from top to bottom: XRPD pattern of Form CS7 before and after being stored under 25 C./60% RH and 40 C./75% RH for 4 weeks and 80 C. for 1 day).

(155) No form change and obvious purity decrease was observed for Form CS7 after being stored at 25 C./60% RH and 40 C./75% RH for 4 weeks and 80 C. for 1 day. It can be seen that Form CS7 has good stability.

(156) Solubility Study of Form CS7

(157) The prepared solid of apabetalone Form CS7 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, 4 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 6.2.

(158) TABLE-US-00024 TABLE 6.2 Solubility Time (mg/mL) (h) SGF FeSSIF 1 0.38 0.19 4 0.42 0.17 24 0.58 0.29

(159) The above results show that Form CS7 of apabetalone has good solubility in SGF and FeSSIF. Polymorph with high solubility is beneficial to increase the blood concentration of drugs in human body and improve the bioavailability of drugs, which is of great significance for drug research.

(160) Hygroscopicity Study of Form CS7

(161) Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS7 of the present disclosure with about 10 mg of samples. The result is listed in Table 6.3.

(162) TABLE-US-00025 TABLE 6.3 Weight gain under 80% Form relative humidity Form CS7 0.79%

(163) The results showed that weight gain of Form CS7 under 80% RH is 0.79%. According to the hygroscopicity criteria, Form CS7 is slightly hygroscopic. The XRPD pattern of Form CS7 after DVS test was shown in FIG. 6E. No form change was observed for Form CS7 before and after DVS test, which indicates that Form CS7 has good humidity stability.

(164) Form CS7 of the disclosure shows low hygroscopicity, which can well avoid the problems such as crystal instability in the process of drug preparation and/or storage, as well as the unprocessability of the preparation caused by external factors such as environmental moisture, which is conducive to the accurate quantitative preparation and later transportation and storage.

Example 7

(165) Preparation of Form CS9

(166) The process of preparing Form CS9 comprises the following steps:

(167) Dissolving step: About 10 mg of apabetalone solid was dissolved in corresponding solvent of Table 7.1, and a clear solution was obtained by filtering.

(168) Precipitation step: The prepared solution was left at room temperature for slow evaporation until solid precipitated. The obtained solid was Form CS9 of Apabetalone.

(169) Said reaction conditions, solvent composition and solvent amount of the preparation method of apabetalone Form CS9 are shown in Table 7.1. The samples 7-a to 7-c were confirmed to be Form CS9 by XRPD.

(170) TABLE-US-00026 TABLE 7.1 Whether to add Sample Weight Solvent Volume polymer or not T No. (mg) (v/v) (mL) (Y/N) ( C.) 7-a 10.0 Tetrahydrofuran/ 2.7 N 25 isopropanol (1:1) 7-b 10.0 Dichloromethane/ 1.7 N 25 isopropanol (4:1) 7-c 10.7 Chloroform/ 1.7 Y 25 acetonitrile (2:1)

(171) Sample 7-b was selected for characterization. The XRPD pattern is substantially as depicted in FIG. 7A, and the XRPD data are listed in Table 7.2. The DSC curve is substantially as depicted in FIG. 7B, which shows the first endothermic peak at around 203 C. The TGA curve is substantially as depicted in FIG. 7C, which shows about 0.9% weight loss when heated to 200 C.

(172) TABLE-US-00027 TABLE 7.2 2 Theta d spcaing Intensity % 3.92 22.55 17.55 5.99 14.75 7.28 7.25 12.19 64.22 7.94 11.13 6.24 9.92 8.91 24.30 12.09 7.32 8.02 12.78 6.93 17.69 13.36 6.63 22.02 13.53 6.54 10.74 14.48 6.12 2.15 16.98 5.22 19.92 20.01 4.44 7.41 20.88 4.26 5.94 21.42 4.15 21.02 21.77 4.08 16.45 22.00 4.04 14.36 22.53 3.95 100.00 23.02 3.86 25.09 23.95 3.72 34.15 24.19 3.68 58.53 24.86 3.58 16.90 25.83 3.45 25.83 26.11 3.41 24.07 26.53 3.36 7.10 27.03 3.30 6.89 27.81 3.21 7.53 28.51 3.13 10.88 30.61 2.92 3.05 31.17 2.87 2.73 32.29 2.77 2.32 32.80 2.73 2.65 33.67 2.66 2.03 36.63 2.45 1.42 37.12 2.42 1.68
Stability Study of Form CS9

(173) Two samples of apabetalone Form CS9 were placed in constant temperature and humidity chambers at 25 C./60% RH and 40 C./75% RH for 10 months in open dishes. Crystalline form of the sample were tested by XRPD and impurity of the sample were tested to check the stability of Form CS9. The XRPD pattern overlay pattern is substantially as depicted in FIG. 7D (from top to bottom: XRPD pattern of Form CS9 before and after being stored under 25 C./60% RH and 40 C./75% RH for 10 months).

(174) No form change and obvious purity decrease was observed for Form CS9 after being stored at 25 C./60% RH and 40 C./75% RH for 10 months. It can be seen that Form CS9 has good stability.

(175) Solubility Study of Form CS9

(176) The prepared solid of apabetalone Form CS9 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, 4 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 7.3.

(177) TABLE-US-00028 TABLE 7.3 Solubility Time (mg/mL) (h) SGF FeSSIF 1 0.30 0.17 4 0.31 0.13 24 0.37 0.20

(178) The above results show that Form CS9 of apabetalone has good solubility in SGF and FeSSIF.

(179) Hygroscopicity Study of Form CS9

(180) Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS9 of the present disclosure with about 10 mg of sample. The result is listed in Table 7.4. The DVS plot of Form CS9 is substantially as depicted in FIG. 7E. The XRPD pattern of Form CS9 after DVS test was shown in FIG. 7F. No form change was observed for Form CS9 before and after DVS test, which indicates that Form CS9 is stable under the influence of humidity.

(181) TABLE-US-00029 TABLE 7.4 Weight gain under 80% Form relative humidity Form CS9 0.18%

(182) The results showed that weight gain of Form CS9 under 80% RH is 0.18%. According to the hygroscopicity criteria, Form CS9 is almost non hygroscopic. Form CS9 of the present disclosure shows low hygroscopicity, which can well avoid the problems such as crystal instability in the process of drug preparation and/or storage, as well as the unprocessability of the preparation caused by external factors such as environmental moisture, which is conducive to the accurate quantitative preparation and later transportation and storage.

Example 8

(183) Preparation of Form CS11 (Method 1)

(184) The process of preparing Form CS11 comprises the following steps:

(185) Dissolving step: about 10 mg of apabetalone solid was dissolved in 1.7 mL of methanol, and filtered to get a clear solution.

(186) Precipitation step: The prepared solution was added to 3 mL of water or 3 mL of water was added to the prepared solution with stirring to obtain solid. Form CS11 was obtained by centrifugation and drying.

(187) Said reaction conditions and amount of methanol and water of the preparation method of apabetalone Form CS11 are shown in Table 8.1. The samples 8-a to 8-b were confirmed to be Form CS11 by XRPD.

(188) TABLE-US-00030 TABLE 8.1 Volume of Volume of Sample Weight methanol water No. (mg) (mL) (mL) Method 8-a 10.0 1.7 3 Antisolvent addition 8-b 10.0 1.7 3 Reverse anti-solvent addition

(189) Sample 8-b was selected for characterization. The XRPD pattern is substantially as depicted in FIG. 8A, and the XRPD data are listed in Table 8.2. The DSC curve is substantially as depicted in FIG. 8B, which shows the first endothermic peak at around 49 C., the second endothermic peak at around 206 C., the first exothermic peak at 208 C. and the third endothermic peak at 230 C. The TGA curve is substantially as depicted in FIG. 8C, which shows about 3.0% weight loss when heated to 100 C.

(190) TABLE-US-00031 TABLE 8.2 2 Theta d spacing Intensity % 4.36 20.25 22.75 7.61 11.61 35.17 7.80 11.33 71.58 8.79 10.06 75.43 9.70 9.12 100.00 10.11 8.75 11.17 13.02 6.80 8.06 13.55 6.54 23.03 15.32 5.79 8.84 15.70 5.64 7.37 16.87 5.26 11.53 17.64 5.03 10.29 18.67 4.75 1.01 19.55 4.54 3.26 20.40 4.35 5.32 20.99 4.23 5.64 21.58 4.12 21.02 22.72 3.91 9.87 23.67 3.76 3.85 25.65 3.47 5.77 26.85 3.32 2.83 27.34 3.26 5.29 30.28 2.95 1.82 32.57 2.75 1.07 34.69 2.59 2.33 35.80 2.51 2.02 36.56 2.46 1.51
Preparation of Form CS11 (Method 2)

(191) The process of preparing Form CS11 comprises the following steps:

(192) Dissolving step: about 10 mg of apabetalone solid was dissolved in corresponding solvent of Table 8.3, and then filtered to get clear solutions.

(193) Precipitation step: The prepared solution was left at room temperature for slow evaporation until solid precipitated. The obtained solid was Form CS11 of Apabetalone.

(194) Said reaction conditions, solvent composition and solvent amount of the preparation method of apabetalone Form CS11 are shown in Table 8.3. The samples 8-c to 8-f were confirmed to be Form CS11 by XRPD. The XRPD pattern of sample 8-d is substantially as depicted in FIG. 8D, and the XRPD data are listed in Table 8.4.

(195) TABLE-US-00032 TABLE 8.3 Sample Weight Solvent Volume T No (mg) (v/v) (mL) ( C.) 8-c 10.4 Chloroform 2.0 25 8-d 10.3 Methanol/acetone (1:1) 1.7 25 8-e 10.1 Methanol/methyl 1.7 25 isobutyl ketone (2:1) 8-f 10.2 Methanol/toluene (4:1) 1.3 25

(196) TABLE-US-00033 TABLE 8.4 2 Theta d spacing Intensity % 4.36 20.26 9.4 7.60 11.64 29.25 7.79 11.34 74.56 8.78 10.07 28.33 9.68 9.14 100 10.13 8.73 8.22 13.00 6.81 6.84 13.54 6.54 13.96 15.29 5.80 8.47 15.68 5.65 5.67 16.83 5.27 7.76 17.65 5.02 4.52 19.48 4.56 3.39 20.37 4.36 4.97 20.97 4.24 3.31 21.60 4.11 8.44 22.65 3.93 9.42 23.08 3.85 2.33 23.67 3.76 3.29 25.65 3.47 0.78 27.34 3.26 2.23 30.18 2.96 1.12 32.46 2.76 0.27 34.67 2.59 0.62 36.15 2.48 0.32
Stability Study of Form CS11

(197) Two samples of apabetalone Form CS11 were placed in constant temperature and humidity chambers at 25 C./60% RH and 40 C./75% RH for 6 weeks in open dishes. Crystalline form of the sample were tested by XRPD and impurity of the sample were tested to check the stability of Form CS11. The XRPD pattern overlay was substantially as depicted in FIG. 8E (from top to bottom: XRPD pattern of Form CS11 before and after being stored under 25 C./60% RH and 40 C./75% RH for 6 weeks).

(198) No form change and obvious purity decrease was observed for Form CS11 after being stored at 25 C./60% RH and 40 C./75% RH for 6 weeks. It can be seen that Form CS11 has good stability.

(199) Solubility Study of Form CS11

(200) The prepared solid of apabetalone Form CS11 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, 4 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 8.5.

(201) TABLE-US-00034 TABLE 8.5 Solubility Time (mg/mL) (h) SGF FeSSIF 1 0.71 0.29 4 0.65 0.26 24 0.65 0.30

(202) The above results show that Form CS11 of apabetalone has good solubility in SGF and FeSSIF. Polymorph with high solubility is beneficial to increase the blood concentration of drugs in human body and improve the bioavailability of drugs, which is of great significance for drug research.

Example 9

(203) Preparation of Form CS4

(204) Form CS4 was obtained by heating Form CS11 of apabetalone to 220 C.

(205) The XRPD pattern of Form CS4 is substantially as depicted in FIG. 9A, and the XRPD data are listed in Table 9.1. The DSC curve is substantially as depicted in FIG. 9B, which shows the first endothermic peak at around 231 C. The TGA curve is substantially as depicted in FIG. 9C, which shows about 1.1% weight loss when heated to 220 C.

(206) TABLE-US-00035 TABLE 9.1 2 Theta d spacing Intensity % 8.48 10.43 6.12 9.11 9.71 53.67 10.29 8.60 26.02 10.81 8.18 10.09 11.64 7.60 14.92 14.17 6.25 7.64 14.45 6.13 45.98 14.84 5.97 7.80 15.34 5.78 2.53 16.38 5.41 4.37 17.06 5.20 2.02 17.72 5.00 1.91 18.32 4.84 2.30 19.25 4.61 6.41 19.53 4.55 12.06 19.80 4.48 2.96 20.73 4.29 2.28 22.60 3.93 2.69 23.51 3.78 83.35 24.23 3.67 100.00 24.99 3.56 19.67 25.47 3.50 7.99 25.89 3.44 4.39 26.32 3.39 35.61 26.97 3.31 6.81 28.30 3.15 5.10 29.95 2.98 2.52 32.05 2.79 5.67 33.73 2.66 2.67 36.61 2.45 3.15 37.68 2.39 1.06
Long-Term Stability Study of Form CS4

(207) Two samples of apabetalone Form CS4 were placed in open dishes in constant temperature and humidity chambers at 25 C./60% RH and 40 C./75% RH for 10 months. Crystalline form of the sample were tested by XRPD and impurity of the sample were tested to check the stability of Form CS4. The stability results are substantially as depicted in FIG. 9D (from top to bottom: XRPD pattern of Form CS4 before and after being stored under 25 C./60% RH and 40 C./75% RH for 10 months) and Table 9.2.

(208) No form change and obvious purity decrease was observed for Form CS4 after being stored at 25 C./60% RH and 40 C./75% RH for 10 months. It can be seen that Form CS4 has good stability.

(209) TABLE-US-00036 TABLE 9.2 Condition 1 week 2 weeks 4 weeks 10 months 25 C./60% RH 99.10 99.06 99.05 99.04 40 C./75% RH 99.10 98.99 99.04 99.02
Solubility Study of Form CS4

(210) The prepared solid of apabetalone Form CS4 was suspended into SGF (simulated gastric fluids) and FeSSIF (fed state simulated intestinal fluids, pH=5.0) to obtain saturated solutions. After being equilibrated for 1 h, 4 h and 24 h, concentrations of the saturated solutions were measured by HPLC. The results are listed in Table 9.3, which indicated that Form CS4 has good solubility.

(211) TABLE-US-00037 TABLE 9.3 Solubility Time (mg/mL) (h) SGF FeSSIF 1 0.30 0.14 4 0.28 0.10 24 0.38 0.16
Hygroscopicity Study of Form CS4

(212) Dynamic vapor sorption (DVS) was applied to test hygroscopicity of Form CS4 of the present disclosure with about 10 mg of sample. The result was listed in Table 9.4. The DVS plot of Form CS4 is as depicted in FIG. 9E.

(213) TABLE-US-00038 TABLE 9.4 Weight gain under 80% Form relative humidity Form CS4 0.12%

(214) The results showed that weight gain of Form CS4 under 80% RH is 0.12%. According to the hygroscopicity criteria, Form CS4 is almost non hygroscopic. The XRPD pattern of Form CS4 after DVS test is shown in FIG. 9F. No form change was observed for Form CS4 before and after DVS test, which indicates that Form CS4 is stable under the influence of humidity.

(215) Form CS4 of the present disclosure shows low hygroscopicity, which can well avoid the problems such as crystal instability in the process of drug preparation and/or storage, as well as the unprocessability of the preparation caused by external factors such as environmental moisture, which is conducive to the accurate quantitative preparation and later transportation and storage.

(216) Mechanical Stability of Form CS4

(217) Certain amount of Form CS4 was placed in a mortar and ground manually for 5 minutes. Crystalline form of the sample was checked by XRPD. The results are shown in FIG. 9G.

(218) The results showed that no form change and obvious crystalline decrease was observed for Form CS4 of apabetalone under certain mechanical stress. From CS4 can maintain stable physical and chemical properties under certain mechanical stress, which is suitable for drug preparation and storage. Grinding of API is usually needed in the process of formulation, and good mechanical stability will reduce the risk of crystallinity decreasing and transformation of solid form of API.

(219) The examples described above are only for illustrating the technical concepts and features of the present disclosure, and intended to make those skilled in the art being able to understand the present disclosure and thereby implement it, and should not be concluded to limit the protective scope of the present disclosure. Any equivalent variations or modifications according to the spirit of the present disclosure should be covered by the protective scope of the present disclosure.