Novel crystal form of deoxycholic acid and preparation method thereof

Abstract

The invention provides polymorphic substances of deoxycholic acid and a preparation method of the polymorphic substances, in particular single pure crystal form G and crystal form H. The deoxycholic acid crystal form G has good stability, the preparation method is simple and easy to operate, the production time is greatly shortened, the efficiency is improved, the energy consumption is reduced, the preparation method is suitable for industrial production, and a better choice is provided for preparation of deoxycholic acid bulk drugs and preparation development.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and relates to polymorphs of deoxycholic acid, especially crystal form G and crystal form H and their preparation methods. Background Art

[0002] Deoxycholic acid is a bile acid lacking a hydroxyl group at C-7, and is a free bile acid derived from cholic acid by losing an oxygen atom. It mainly exists in the form of combination with taurine and glycine in bile. Deoxycholic acid has the functions of promoting bile secretion, regulating immunity, protecting the liver, anti-inflammatory, antioxidant, etc., and is used for preparing drugs for reducing cholesterol, helping fat digestion and absorption, preventing gallstones, treating fatty liver, treating cholestatic hepatitis, etc., or for removing local fat, etc., and has wide applications in the fields of pharmaceuticals and cosmetics.

[0003] Drug polymorphs generally refer to different arrangements of chemical drug molecules, and generally represent the existence forms of drug raw materials in the solid state. The same drug can exist in multiple polymorphic states, and different polymorphs can have different chemical and physical properties, including melting point, chemical reactivity, apparent solubility, dissolution rate, optical and mechanical properties, vapor pressure and density.

[0004] Deoxycholic acid has obvious polymorphic phenomena, and its crystal form and stability directly affect the processing and production of drug raw materials and preparations, and affect the quality, safety and effectiveness of drug preparations. CN103906517A has disclosed four crystal forms A, B, C and D of deoxycholic acid. The inventors of the present invention have verified through experiments that crystal form A can be prepared by pure water pulping, but the post-treatment is inconvenient, the preparation process is very slow, which is disadvantageous for industrial production; while crystal forms B, C and D cannot be repeatedly obtained, and the reproducibility is poor; under most conditions, the deoxycholic acid obtained is a mixture of different crystal forms, and the properties are unstable and prone to crystal transformation.

[0005] In order to meet the requirements of the research and development of deoxycholic acid drug raw materials and preparations, it is necessary to fully study the crystal form of the drug raw materials, drug stability, process controllability, etc., and develop deoxycholic acid products suitable for the application of drug raw materials and preparations. Summary of the Invention

[0006] The purpose of the present invention is to provide polymorphs of deoxycholic acid, especially single pure deoxycholic acid crystal forms G and H and their preparation methods, and crystal form G is further preferred. The production of the above crystal forms provided by the present invention is more convenient, significantly shortens the production time, saves the consumption of manpower and material resources, improves the production efficiency, and has good stability, which provides great convenience for the production and storage of deoxycholic acid drug raw materials, and at the same time provides better guarantee for the stability and safety of drug raw materials and preparations.

[0007] According to the first aspect of the present invention, crystal form G of deoxycholic acid is provided.

[0008] The X-ray powder diffraction pattern of crystalline form G has characteristic peaks at the following 2θ values: 6.80 ± 0.2°, 12.14 ± 0.2°, 13.63 ± 0.2°, 13.98 ± 0.2°, 18.29 ± 0.2° and 18.95 ± 0.2°.

[0009] In some embodiments, the X-ray powder diffraction pattern of crystalline form G has characteristic peaks at the following 2θ values: 6.80 ± 0.2°, 7.38 ± 0.2°, 9.45 ± 0.2°, 12.14 ± 0.2°, 13.63 ± 0.2°, 13.98 ± 0.2°, 18.29 ± 0.2°, 18.95 ± 0.2° and 22.37 ± 0.2°.

[0010] In some embodiments of the present invention, the X-ray powder diffraction pattern of crystalline form G has characteristic peaks at the 2θ values shown in Table 1 below:

[0011] Table 1

[0012] Serial number 2θ ± 0.2° d value Relative intensity Serial number 2θ ± 0.2° d value Relative intensity 1 6.80 12.98 43.1 7 15.14 5.85 15.3 2 7.38 11.98 31.9 8 17.25 5.14 10.1 3 9.45 9.35 7.5 9 18.29 4.85 100.0 4 12.14 7.28 37.4 10 18.95 4.68 35.4 5 13.63 6.49 52.0 11 22.37 3.99 27.8 6 13.98 6.33 59.4 12 24.84 3.58 9.6

[0013] In some embodiments, the X-ray powder diffraction pattern of crystalline form G is substantially as Figure 3 characterized.

[0014] In some embodiments, the DSC pattern of crystalline form G has an endothermic peak in the range of 188 ± 2 °C.

[0015] In some embodiments, the DSC pattern of crystalline form G is substantially as Figure 4 characterized.

[0016] In some embodiments, the preparation method of crystalline form G includes: heating and dissolving the crude deoxycholic acid in methanol, adding water, cooling for crystallization, filtering, and drying to obtain crystalline form G. Preferably, the heating temperature is 60 °C; preferably, the weight-to-volume ratio of the crude deoxycholic acid to methanol is 1 g: 8-10 ml, and the volume ratio of methanol to water is 4:1; preferably, seed crystal G is added before cooling for crystallization.

[0017] In the second aspect of the present invention, crystalline form H of deoxycholic acid is provided.

[0018] The X-ray powder diffraction pattern of crystalline form H has characteristic peaks at the following 2θ values: 9.80 ± 0.2°, 13.07 ± 0.2°, 17.59 ± 0.2°, 20.68 ± 0.2°, 22.93 ± 0.2° and 23.85 ± 0.2°.

[0019] In some embodiments, the X-ray powder diffraction pattern of Form H has characteristic peaks at the following 2θ values: 9.80 ± 0.2°, 11.75 ± 0.2°, 13.07 ± 0.2°, 17.59 ± 0.2°, 19.14 ± 0.2°, 20.68 ± 0.2°, 22.93 ± 0.2°, and 23.85 ± 0.2°.

[0020] In some embodiments of the present invention, the X-ray powder diffraction pattern of Form H has characteristic peaks at the 2θ values shown in Table 2 below:

[0021] Table 2

[0022]

[0023]

[0024] In some embodiments, the X-ray powder diffraction pattern of Form H is substantially as Figure 5 characterized.

[0025] In some embodiments, the DSC pattern of Form H has an endothermic peak in the range of 97 ± 2 °C.

[0026] In some embodiments, the DSC pattern of Form H is substantially as Figure 6 characterized.

[0027] In some embodiments, Form H is a dimethyl sulfoxide solvate.

[0028] In some embodiments, the preparation method of Form H includes: dissolving crude deoxycholic acid in DMSO, cooling and crystallizing, filtering, and drying to obtain Form H. Preferably, the weight-to-volume ratio of crude deoxycholic acid to DMSO is 1 g:5 - 10 ml.

[0029] The third aspect of the present invention provides a pharmaceutical composition comprising at least one of Form G and Form H of the present invention, and optionally a pharmaceutically acceptable carrier.

[0030] The fourth aspect of the present invention further provides the use of a crystalline form G or crystalline form H of deoxycholic acid, or the pharmaceutical composition as described above, in the preparation of a drug for preventing and / or treating symptoms or diseases related to local fat deposition.

[0031] The fifth aspect of the present invention further provides the use of a crystalline form G or crystalline form H of deoxycholic acid, or the pharmaceutical composition as described above, in the preparation of a drug for non-surgical removal of local fat deposition.

[0032] The present invention provides new crystal forms of deoxycholic acid, including crystal form G and crystal form H of deoxycholic acid. Both crystal form G and crystal form H have good reproducibility. Crystal form G has excellent stability, and the preparation method is simple and easy to operate, greatly shortening the production time, improving the efficiency, reducing the energy consumption, and being suitable for industrial production applications, providing a favorable guarantee for the quality stability and safety of deoxycholic acid raw materials and preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 and Figure 2 are respectively the XRPD pattern and DSC graph of deoxycholic acid crystal form A;

[0034] Figure 3 and Figure 4 are respectively the XRPD pattern and DSC graph of deoxycholic acid crystal form G;

[0035] Figure 5 and Figure 6 are respectively the XRPD pattern and DSC graph of deoxycholic acid crystal form H;

[0036] Figures 7 - 26 are respectively the DSC graphs of deoxycholic acid polymorphs prepared under different conditions. DETAILED DESCRIPTION OF THE INVENTION

[0037] As mentioned above, deoxycholic acid has obvious polymorphism, and its crystal form and stability directly affect the processing and production of raw materials and preparations of drugs, and affect the quality, safety and effectiveness of pharmaceutical preparations.

[0038] The prior art CN103906517A discloses four crystal forms of deoxycholic acid. Among them, the A-type polymorph is an anhydrous polymorph, and the XRPD pattern has characteristic peaks at 2θ values of 8.9°, 10.7°, 14.0°, 15.0°, 16.2° and 19.1°; the DSC curve has an endothermic peak in the range of 174 ± 2 °C; the B-type polymorph is an anhydrous polymorph, and the XRPD pattern has characteristic peaks at 2θ values of 6.7°, 7.3°, 7.4°, 8.4°, 9.3°, 11.2°, 12.9°, 13.9°, 14.4°, 14.6°, 14.8°, 15.8°, 16.0°, 16.9° and 17.8°; the DSC curve has an endothermic peak in the range of 135 ± 2 °C; the C-type polymorph is a hydrate, and the XRPD pattern has characteristic peaks at 2θ values of 6.6°, 7.3°, 7.4°, 9.6°, 9.9°, 12.6°, 13.0°, 13.2°, 13.9°, 14.2°, 15.1°, 15.6°, 15.8°, 16.4°, 17.0°, 17.1° and 17.6°; the DSC curve has a broad transition peak below 100 °C; the D-type polymorph is an anhydrous polymorph, and the XRPD pattern has characteristic peaks at 2θ values of 7.0°, 7.4°, 10.0°, 14.2°, 15.3°, 15.8°, 16.6° and 17.3°; the DSC curve has an endothermic peak in the range of 156 ± 2 °C.

[0039] The patent application CN106146593A of the present applicant discloses a preparation method of deoxycholic acid, which includes pulping crude deoxycholic acid with pure water at 60 - 80 °C for 4 - 8 h, filtering, drying by suction, and drying to obtain pure deoxycholic acid. The product is confirmed to be crystal form A. However, during the preparation process of crystal form A, due to the viscosity of the pulp, the filtration step is very slow, and manual assistance is still required for compression under vacuum filtration conditions to promote filtration. The washing and drying processes after filtration also take a long time. For example, in industrial production, to complete the purification of 500 g of crude product according to this method, filtration takes about 4 hours and the drying process takes about 24 hours. The time spent in the purification process of the crude product is very long, and there are disadvantages such as high energy consumption and low efficiency.

[0040] During the process of the present inventor improving the production and purification process of crystal form A, it was found that by changing the post-treatment conditions, the obtained deoxycholic acid compounds are all mixtures of different crystal forms, and it is difficult to obtain a single pure crystal form A; and crystal form A may undergo a crystal transformation phenomenon under certain specific conditions, which will affect the production and storage of the active pharmaceutical ingredient and the preparation, and is an important problem that needs to be solved by pharmaceutical production enterprises.

[0041] Based on the prior art and existing problems, the present inventors further conducted in-depth research on the polymorphs of deoxycholic acid through different methods (including evaporation method, heating and cooling method, anti-solvent addition method, suspension stirring method, etc.) or different solvent systems, and confirmed that polymorph A can be obtained under specific conditions, while the reproducibility of polymorphs B, C, and D is poor and single polymorphs cannot be obtained; the present invention also confirmed that most of the deoxycholic acid compounds obtained under different conditions are mixed crystal substances and it is difficult to obtain single pure crystals; unexpectedly, however, we found that new single pure polymorphs G and H were obtained. By further investigating the properties of the deoxycholic acid polymorphs, it was found that under different conditions, the stability of polymorph G is equivalent to or better than that of polymorph A, and the preparation process of polymorph G is more convenient, the treatment process is faster, the efficiency is higher, significantly saving production time and cost, and being more conducive to the industrial production of APIs and preparations; polymorph H has a relatively low melting point and good stability.

[0042] Polymorph G provided by the present invention is an anhydrate, and the XRPD has characteristic peaks at the following 2θ values: 6.80±0.2°, 12.14±0.2°, 13.63±0.2°, 13.98±0.2°, 18.29±0.2° and 18.95±0.2°. The DSC diagram of polymorph G has an endothermic peak in the range of 188±2°C. Polymorph H provided by the present invention is a dimethyl sulfoxide solvate, and the XRPD diagram has characteristic peaks at the following 2θ values: 9.80±0.2°, 13.07±0.2°, 17.59±0.2°, 20.68±0.2°, 22.93±0.2° and 23.85±0.2°. The DSC diagram of polymorph H has an endothermic peak in the range of 97±2°C.

[0043] The solvate referred to in the present invention means a substance (solvate) formed by a compound and a solvent after crystallization. During the contact process between a compound or a drug molecule and a solvent molecule, it is inevitable that due to external and internal condition factors, the solvent molecule and the compound molecule form a co-crystal and remain in the solid substance. Solvent types that are likely to form solvates with organic compounds are water, methanol, benzene, ethanol, ether, aromatic hydrocarbons, heteroaromatic hydrocarbons, etc.

[0044] When the terms "about" or "approximately" are used in combination with a numerical quantity or range, it means slightly greater than or slightly less than the said numerical quantity or range, with a deviation of ±10% from the end values of the said numerical quantity or range.

[0045] The present invention will be described in detail below through examples. It should be understood that the methods in the examples are only for illustrative purposes and do not constitute any limitation to the present invention. Unless otherwise specified, the materials and instruments used in the examples are conventional or commonly commercially available materials and instruments known in the art. Unless otherwise specified, the experimental methods used are conventional methods known in the art.

[0046] Materials, Instruments and Methods:

[0047] Crude deoxycholic acid: HPLC purity 97.80%

[0048] XRPD (X-ray powder diffraction): Instrument: Bruker D8 ADVANCE X-ray diffractometer; XRPD parameters: X-ray tube: Cu, kα, X-ray tube voltage: 40 kV, X-ray tube current: 40 mA; Scanning range: 3 - 45 deg; Step size: 0.02 deg; Step time: 0.12 s

[0049] DSC (Differential scanning calorimetry): Instrument: METTLER TOLEDO DSC3+ differential scanning calorimeter; DSC parameters: Temperature range: 25 - 300 °C; Heating rate: 10 °C / min; Nitrogen purge gas: 50 ml / min.

[0050] Example 1. Preparation of deoxycholic acid polymorphs

[0051] Add 1.0 g of crude deoxycholic acid to 10 ml of water, slurry at 70 °C for 3 hours, cool to room temperature for crystallization, filter, wash with a small amount of water, collect the solid, and dry in vacuo at 40 °C for about 12 h to obtain a white powdery solid compound.

[0052] The obtained compound is crystalline form A. The XRPD pattern is shown in Figure 1 . The DSC pattern is basically as Figure 2 shown, with an endothermic peak at about 173.6 °C. Crystalline form A is an anhydrate.

[0053] Example 2. Preparation of deoxycholic acid polymorphs

[0054] Add 1.0 g of crude deoxycholic acid to 10 ml of methanol, stir to dissolve at 60 °C, add 2.5 ml of purified water, cool to about 5 °C for crystallization, filter, wash with a small amount of methanol / water (4:1) mixed solvent, and dry the solid in vacuo at 40 °C for 3 h to obtain a white crystalline compound.

[0055] The obtained compound is crystalline form G. The XRPD pattern is shown in Figure 3 , and the diffraction angle data are basically as shown in Table 1. The DSC pattern is basically as Figure 4 shown, with a single strong endothermic peak at about 188.0 °C. Crystalline form G is an anhydrate.

[0056] Example 3. Preparation of deoxycholic acid polymorphs

[0057] 1.0 g of crude deoxycholic acid was added to 5 ml of dimethyl sulfoxide solvent and dissolved at room temperature. After standing for crystallization for 10 days, a solid was obtained, which was filtered, washed, and dried in vacuo at 40 °C to obtain 0.7 g of a white powdery solid compound with an HPLC purity of 99.90%.

[0058] The obtained compound was crystal form H. The XRPD pattern is shown in Figure 5 , and the diffraction angle data are basically as shown in Table 2. The DSC pattern is basically as Figure 6 shown, with a relatively broad endothermic peak in the range of about 97 °C. Crystal form H is a dimethyl sulfoxide solvate.

[0059] Example 4. Preparation of deoxycholic acid polymorphs

[0060] 1.0 g of crude deoxycholic acid was added to 10 ml of water and stirred at room temperature for 3 hours for pulping, then filtered and washed with a small amount of water. The solid was collected and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern of the obtained compound is as Figure 7 shown, and it is speculated to be a mixed crystal of crystal forms A and D.

[0061] Example 5. Preparation of deoxycholic acid polymorphs

[0062] 1.0 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of methanol and purified water (1:1) and stirred at room temperature for 3 hours for pulping, then filtered and the solid was collected and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern of the obtained compound is as Figure 8 shown, and it is speculated to be a mixed crystal of crystal forms A and D.

[0063] Example 6. Preparation of deoxycholic acid polymorphs

[0064] 1.0 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of methanol and ethyl acetate (1:1), heated to dissolve, allowed to stand for crystallization at room temperature, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 9 shown. The obtained compound is speculated to be a mixed crystal of crystal forms A and D.

[0065] Example 7. Preparation of deoxycholic acid polymorphs

[0066] 500 mg of crude deoxycholic acid was added to 10 ml of a mixed solvent of methanol and acetonitrile (1:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as shown in Figure 10. The obtained compound is speculated to be a mixed crystal of crystal forms A, D, and another new crystal form (labeled as E).

[0067] Example 8. Preparation of deoxycholic acid polymorphs

[0068] 1 g of crude deoxycholic acid was added to 6 ml of a mixed solvent of ethanol and water (5:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as shown in Figure 11 shown. The obtained compound is presumably a mixed crystal of crystal forms A and D.

[0069] Example 9. Preparation of deoxycholic acid polymorphs

[0070] 1 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of ethanol and acetonitrile (1:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as shown in Figure 12 shown. The obtained compound is presumably a mixed crystal of crystal forms A, B and D.

[0071] Example 10. Preparation of deoxycholic acid polymorphs

[0072] 1 g of crude deoxycholic acid was added to 35 ml of a mixed solvent of ethanol and acetonitrile (1:5), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as shown in Figure 13 shown. The obtained compound is presumably a mixed crystal of crystal forms A, D and another new crystal form (labeled as E).

[0073] Example 11. Preparation of deoxycholic acid polymorphs

[0074] 1 g of crude deoxycholic acid was added to 40 ml of a mixed solvent of ethanol and methyl tert-butyl ether (1:5), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as shown in Figure 14 shown. The obtained compound is presumably a mixed crystal of crystal forms A, D and another new crystal form (labeled as F).

[0075] Example 12. Preparation of deoxycholic acid polymorphs

[0076] 1 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of tetrahydrofuran and ethyl acetate (1:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as shown in Figure 15 shown. The obtained compound is presumably a mixed crystal of crystal forms A and D.

[0077] Example 13. Preparation of deoxycholic acid polymorphs

[0078] 1 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of tetrahydrofuran and acetonitrile (1:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as shown in Figure 16As shown. The obtained compound is presumably a mixed crystal of crystal forms A, B, and D.

[0079] Example 14. Preparation of deoxycholic acid polymorphs

[0080] 1 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of tetrahydrofuran and acetonitrile (1:5), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 17 shown. The obtained compound is presumably a mixed crystal of crystal forms A, D, and another new crystal form (designated as E).

[0081] Example 15. Preparation of deoxycholic acid polymorphs

[0082] 1 g of crude deoxycholic acid was added to 30 ml of a mixed solvent of tetrahydrofuran and methyl tert-butyl ether (1:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 18 shown. The obtained compound is presumably a mixed crystal of crystal forms A, D, and another new crystal form (designated as F).

[0083] Example 16. Preparation of deoxycholic acid polymorphs

[0084] 500 mg of crude deoxycholic acid was added to 25 ml of acetone, heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 19 shown. The obtained compound is presumably a mixed crystal of crystal forms A, D, and another new crystal form (designated as E).

[0085] Example 17. Preparation of deoxycholic acid polymorphs

[0086] 1 g of crude deoxycholic acid was added to 15 ml of ethyl acetate, heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 20 shown. The obtained compound is presumably a mixed crystal of crystal forms A and D.

[0087] Example 18. Preparation of deoxycholic acid polymorphs

[0088] 1 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of methanol and toluene (1:5), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 21 shown. The obtained compound is presumably a mixed crystal of crystal forms A, D, and an unknown crystal form (designated as F).

[0089] Example 19. Preparation of deoxycholic acid polymorphs

[0090] 1 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of dioxane and water (4:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 22 shown. The obtained compound was presumed to be a mixed crystal of crystal form A and an unknown crystal form (designated as E).

[0091] Example 20. Preparation of deoxycholic acid polymorphs

[0092] 1 g of crude deoxycholic acid was added to 10 ml of a mixed solvent of dioxane and water (8:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 23 shown. The obtained compound was presumed to be a mixed crystal of crystal forms A, D and an unknown crystal form (designated as E).

[0093] Example 21. Preparation of deoxycholic acid polymorphs

[0094] 1 g of crude deoxycholic acid was added to 20 ml of a mixed solvent of dioxane and acetonitrile (1:1), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 24 shown. The obtained compound was presumed to be a mixed crystal of crystal form A and another new crystal form (designated as E).

[0095] Example 22. Preparation of deoxycholic acid polymorphs

[0096] 500 mg of crude deoxycholic acid was added to 40 ml of a mixed solvent of dioxane and acetonitrile (1:5), heated to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 25 shown. The obtained compound was presumed to be a mixed crystal of crystal forms A, D and another new crystal form (designated as E).

[0097] Example 23. Preparation of deoxycholic acid polymorphs

[0098] 1 g of crude deoxycholic acid was added to 50 ml of a mixed solvent of methanol and dichloromethane (1:5), heated with stirring to dissolve, cooled and allowed to stand for crystallization, filtered, and dried in vacuo at 40 °C to obtain a white powdery solid compound. The DSC pattern is as Figure 26 shown. The obtained compound was presumed to be a mixed crystal of crystal forms A and D.

[0099] Example 24. Preparation of deoxycholic acid polymorphs

[0100] 50 g of crude deoxycholic acid was added to 500 ml of water, and the mixture was slurried at 70 °C for 6 hours, filtered and washed with a small amount of water (about 4 h). The solid was collected and dried in vacuo at 40 °C for about 6 h. The compound was not completely dry (water content about 5%); it was further dried for 18 h to obtain 45.2 g of a white powdery compound with an HPLC purity of 99.90% and a water content of less than 1.0%. The obtained compound was crystalline form A.

[0101] Example 25. Preparation of deoxycholic acid polymorphs

[0102] 50 g of crude deoxycholic acid was added to 400 ml of methanol, and the mixture was stirred and dissolved at 60 °C. 100 ml of purified water was added, and seed crystal A was added. The temperature was lowered to about 5 °C, and crystallization was carried out for 6 h. The mixture was filtered and washed with a small amount of ice methanol / water (4:1) (about 1.5 h). The solid was dried in vacuo at 40 °C for 6 h to obtain 42.6 g of a white crystalline compound with an HPLC purity of 99.97% and a water content of less than 1.0%. The obtained compound was crystalline form G.

[0103] Example 26. Physicochemical properties of deoxycholic acid polymorphs

[0104]

[0105] Example 27. Stability study of deoxycholic acid polymorphs

[0106] Samples of crystalline forms A, G, and H of deoxycholic acid were placed separately in an open and flat manner to investigate the physical stability of the samples under high temperature (60 °C), high humidity (RH 92.5%), and light conditions. At 0 days and 30 days, a certain amount of samples were taken for DSC and XRPD tests to evaluate the physical stability, and a certain amount of samples were taken for HPLC tests to evaluate the chemical stability. The results are shown in Table 3.

[0107] Table 3. Results of stability study

[0108]

[0109] The experimental results showed that crystalline forms A, G, and H had good chemical stability under high temperature, high humidity, and light conditions, and HPLC showed that the content did not change significantly. Crystalline forms A and H could be converted into a mixed crystal of crystalline forms A and D after being placed under high humidity conditions for 30 days. Crystalline form G had excellent stability.

[0110] In summary, through the research on the crystal forms of deoxycholic acid, the present inventors obtained single pure crystal forms G and H of deoxycholic acid. In addition, new crystal forms E (melting point about 142±2 °C) and F (melting point about 165±2 °C) were also discovered, but they all existed in mixed crystal compounds and single pure crystal forms were not obtained. Among them, deoxycholic acid crystal form G has good physical and chemical stability, the preparation method is simple, the production time is significantly shortened, the consumption of manpower and energy is reduced, and it is suitable for industrial production, providing a better choice for the preparation of deoxycholic acid bulk drugs and formulation development.

[0111] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them; although the embodiments have described the present invention in detail, those of ordinary skill in the art should understand that based on the disclosed content of the present invention, the technical solutions recorded in the foregoing embodiments may be modified, or some of the technical features may be equivalently replaced; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the present invention and still fall within the protection scope of the present invention.

Claims

1. A deoxycholic acid compound, the compound being selected from crystalline form G and crystalline form H; characterized in that: The X-ray powder diffraction pattern of the crystalline form G has characteristic peaks at the following 2θ values: 6.80 ± 0.2°, 12.14 ± 0.2°, 13.63 ± 0.2°, 13.98 ± 0.2°, 18.29 ± 0.2°, and 18.95 ± 0.2°; The X-ray powder diffraction pattern of the crystalline form H has characteristic peaks at the following 2θ values: 9.80 ± 0.2°, 13.07 ± 0.2°, 17.59 ± 0.2°, 20.68 ± 0.2°, 22.93 ± 0.2°, and 23.85 ± 0.2°.

2. The deoxycholic acid compound according to claim 1, characterized in that: The X-ray powder diffraction pattern of the crystalline form G has characteristic peaks at the following 2θ values: 6.80 ± 0.2°, 7.38 ± 0.2°, 9.45 ± 0.2°, 12.14 ± 0.2°, 13.63 ± 0.2°, 13.98 ± 0.2°, 18.29 ± 0.2°, 18.95 ± 0.2°, and 22.37 ± 0.2°; The X-ray powder diffraction pattern of the crystalline form H has characteristic peaks at the following 2θ values: 9.80 ± 0.2°, 11.75 ± 0.2°, 13.07 ± 0.2°, 17.59 ± 0.2°, 19.14 ± 0.2°, 20.68 ± 0.2°, 22.93 ± 0.2°, and 23.85 ± 0.2°.

3. The deoxycholic acid compound according to claim 1, wherein The X-ray powder diffraction pattern of the crystalline form G is substantially characterized as shown in Figure 3; the X-ray powder diffraction pattern of the crystalline form H is substantially characterized as shown in Figure 5.

4. The deoxycholic acid compound according to claim 1, wherein The DSC curve of the crystalline form G has an endothermic peak in the range of 188 ± 2 °C; the DSC curve of the crystalline form H has an endothermic peak in the range of 97 ± 2 °C.

5. The deoxycholic acid compound according to claim 1, wherein The DSC curve of the crystalline form G is substantially characterized as shown in Figure 4; the DSC curve of the crystalline form H is substantially characterized as shown in Figure 6.

6. A method for preparing the deoxycholic acid compound as described in claim 1, characterized in that, The compound is crystalline form G, and the method includes: heating and dissolving the crude deoxycholic acid in methanol, adding water, and cooling for crystallization to obtain crystalline form G; preferably, the heating temperature is 60 °C; preferably, the volume ratio of methanol to water is 4:1; preferably, seed crystal G is added before cooling for crystallization.

7. A method for preparing the deoxycholic acid compound as claimed in claim 1, characterized in that, The compound is crystalline form H, and the method includes: dissolving the crude deoxycholic acid in DMSO at room temperature, cooling for crystallization to obtain crystalline form H.

8. A pharmaceutical composition comprising the deoxycholic acid compound according to any one of claims 1 to 5, and optionally a pharmaceutically acceptable carrier.

9. Use of the deoxycholic acid compound according to any one of claims 1 to 5, or the pharmaceutical composition according to claim 8, in the preparation of a drug for preventing and / or treating symptoms or diseases related to local fat deposition.

10. Use of the deoxycholic acid compound according to any one of claims 1 to 5, or the pharmaceutical composition according to claim 8, in the preparation of a drug for non-surgical removal of local fat deposition.

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