High-purity deoxycholic acid and preparation method thereof

Abstract

The invention discloses high-purity deoxycholic acid and a preparation method thereof.The method comprises the steps that S1, industrial-grade deoxycholic acid serves as a raw material and is dissolved in water, the pH of a system is adjusted to be alkaline, and a solution A is obtained; s2, adding an extraction agent into the solution A for extraction and liquid separation so as to remove fat-soluble impurities and macromolecular organic impurities, then adding a removal agent and an adsorbent into an extracted water phase liquid so as to respectively remove heavy metal ions and small molecular pigments, and then filtering to remove the adsorbent so as to obtain a filtrate A; s3, filtering the filtrate A for the second time to obtain filtrate B, then adjusting the pH of the filtrate B to be acidic, filtering again, and collecting a filter cake; s4, adding an organic solvent into the filter cake, heating until the organic solvent is completely dissolved, and then filtering while the filter cake is hot to obtain filtrate C; and S5, dropwise adding water into the filtrate C until turbidity appears, then cooling and crystallizing, continuously filtering, collecting crystals, and carrying out vacuum drying to obtain the high-purity deoxycholic acid.

Description

Technical Field

[0001] This invention relates to the field of deoxycholic acid purification technology, specifically to a high-purity deoxycholic acid and its preparation method. Background Technology

[0002] Deoxycholic acid is an important pharmaceutical chemical product, widely used in the preparation of topical lipolysis drugs. In 2015, the U.S. Food and Drug Administration (FDA) approved deoxycholic acid injection as the world's first topical lipolysis drug for cosmetic purposes, bringing it significant attention in the pharmaceutical and cosmetic fields.

[0003] Currently, domestic and international literature and patent reports mainly report two methods for preparing deoxycholic acid. One method uses animal bile acids as starting materials and produces deoxycholic acid through a redox reaction. The other method uses 9-hydroxyandrostenedione, cortisone, and hydrocortisone as starting materials and produces deoxycholic acid through more than ten reaction steps. However, the second method has a long synthesis process and low yield, and is still far from industrial application. Currently, deoxycholic acid sold in the domestic market is mainly extracted from the bile of animals such as cattle and sheep. It contains many impurities, such as bile acids, small molecule pigments, lipid-soluble impurities, particulate impurities, large molecule pollutants, and heavy metals such as arsenic and lead, which cannot yet meet the quality requirements for drug research. Summary of the Invention

[0004] The purpose of this invention is to provide a method for preparing high-purity deoxycholic acid, which aims to solve the problem that existing deoxycholic acid contains many impurities and high levels of arsenic and heavy metal ions, thus failing to meet drug quality requirements.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solution:

[0006] This invention provides a method for preparing high-purity deoxycholic acid, which includes the following steps:

[0007] S1. Using industrial-grade deoxycholic acid as raw material, dissolve it in water and adjust the pH of the system to be alkaline to obtain solution A;

[0008] S2. Add an extractant to the solution A for extraction and separation to remove lipid-soluble impurities and macromolecular organic impurities. Then add a desiccant and an adsorbent to the aqueous phase after extraction to remove arsenic, heavy metal ions and small molecule pigments respectively. Then filter to remove the adsorbent to obtain filtrate A.

[0009] S3. Filter the filtrate A a second time to obtain filtrate B, then adjust the pH of filtrate B to acidic, filter it again, and collect the filter cake.

[0010] S4. Add an organic solvent to the filter cake and heat until completely dissolved, then filter while hot to obtain filtrate C;

[0011] S5. Add water dropwise to the filtrate C until it becomes turbid, then cool it to crystallize, continue filtering, collect the crystals, and vacuum dry to obtain high-purity deoxycholic acid.

[0012] Furthermore, a method for preparing high-purity deoxycholic acid: in step S1, the purity of deoxycholic acid is 90-97%; the mass-to-volume ratio of deoxycholic acid to water is 0.05-0.1 g / ml; and the pH of the system is adjusted to 8-9 using sodium hydroxide solution.

[0013] Furthermore, a method for preparing high-purity deoxycholic acid: the extractant in step S2 is selected as dichloromethane or ethyl acetate; the volume ratio of the extractant to water is 1:(1-5).

[0014] Furthermore, a method for preparing high-purity deoxycholic acid: the removal agent is sodium sulfide, and the adsorbent is activated carbon.

[0015] Furthermore, a method for preparing high-purity deoxycholic acid: the amount of the removal agent added is 0.01-0.2 wt% of the mass of industrial-grade deoxycholic acid, and the amount of the adsorbent added is 0.5-5.0% of the mass of industrial-grade deoxycholic acid.

[0016] Furthermore, a method for preparing high-purity deoxycholic acid: the amount of the removal agent added is 0.05-0.1 wt% of the mass of industrial-grade deoxycholic acid; the amount of the adsorbent added is 1.0-3.0% of the mass of industrial-grade deoxycholic acid.

[0017] Furthermore, a method for preparing high-purity deoxycholic acid: Step S3, the filtrate A is filtered a second time using a mixed fiber resin microporous membrane with a pore size of 0.45 μm to obtain filtrate B, and then the pH of filtrate B is adjusted to 1-2 with hydrochloric acid, filtered again, and the filter cake is collected.

[0018] Furthermore, a method for preparing high-purity deoxycholic acid: Step S4, add ethanol to the filter cake and heat to 55-65°C. After complete dissolution, pass the hot solution through a PTFE microfiltration membrane with a pore size of 0.45 μm to obtain filtrate C.

[0019] Furthermore, a method for preparing high-purity deoxycholic acid: Step S5, add deionized water dropwise to the filtrate C until turbidity appears, then slowly cool to 3-7℃ at a cooling rate of 0.5-1.0℃ / min to crystallize, then filter, collect the crystals, and dry them in a vacuum oven at a temperature of 45-60℃ and a vacuum degree of 0.5-2.0KPa to obtain high-purity deoxycholic acid.

[0020] The present invention also provides a high-purity deoxycholic acid, which is prepared by the above-mentioned preparation method. The high-purity deoxycholic acid has a purity of ≥99%, a specific rotation of +54.0 to +57.0, a melting point of 173 to 174°C, an arsenic content of <0.0001%, and a heavy metal content of <0.0005%.

[0021] Specifically, the method for preparing high-purity deoxycholic acid of the present invention first uses solvent extraction to remove lipid-soluble impurities and macromolecular organic impurities, then uses sodium sulfide to remove heavy metals such as arsenic and lead, then uses activated carbon to adsorb small molecule pigments, acidifies the obtained deoxycholic acid, then dissolves it in ethanol, removes particulate impurities by microfiltration membrane heat filtration, adds deionized water until turbidity appears, crystallizes by gradient cooling, and vacuum dries it to obtain high-purity deoxycholic acid with extremely low arsenic and heavy metal content and a purity ≥99%.

[0022] The beneficial effects of this invention are:

[0023] The method for preparing high-purity deoxycholic acid proposed in this invention belongs to the category of deoxycholic acid purification methods. It can purify industrial-grade deoxycholic acid with a purity of only 90-97% extracted from bovine bile into high-purity deoxycholic acid with a purity greater than 99%. Furthermore, it can ensure that the arsenic content of the obtained deoxycholic acid is <0.0001% and the heavy metal content is <0.0005%, thus solving the problems of high impurities and high content of heavy metal ions such as arsenic in existing deoxycholic acid, thereby meeting the quality requirements of pharmaceuticals. Detailed Implementation

[0024] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0025] Example 1

[0026] This embodiment 1 provides a method for preparing high-purity deoxycholic acid, which includes the following specific steps:

[0027] S1. Add 1500 ml of deionized water to a reaction apparatus equipped with a stirrer and a temperature controller. Then, under stirring conditions, add 100.0 g of industrial-grade deoxycholic acid (light pink, purity 95.5%, specific rotation +52.4, melting point 166.9~171.1℃, arsenic content 0.0003%, heavy metal content 0.002%). Continue to add 10% sodium hydroxide solution dropwise to the reaction apparatus until the pH of the system is 8~9 to obtain solution A.

[0028] S2. Add 500 ml of dichloromethane to the above solution A for extraction. After stirring and mixing, separate the liquid and repeat the extraction and separation once to remove lipid-soluble impurities and macromolecular organic impurities in deoxycholic acid. Then, add 0.05 g of sodium sulfide (removing agent) and 3.0 g of pre-activated activated carbon (adsorbent) to the aqueous phase after extraction and stir at 60 °C for 2 hours to remove heavy metal ions and small molecule pigments respectively. Then, filter with medium-speed filter paper to remove activated carbon to obtain filtrate A.

[0029] S3. The above filtrate A is filtered a second time using a mixed fiber resin microporous filter membrane with a pore size of 0.45μm to obtain filtrate B. Then, hydrochloric acid is added dropwise to filtrate B to adjust the pH of filtrate B to 1-2, and the filter cake is collected after filtration.

[0030] S4. Add 800 ml of ethanol to the obtained filter cake and heat to 60°C. After complete dissolution, pass the hot solution through a PTFE microfiltration membrane with a pore size of 0.45 μm to obtain filtrate C.

[0031] S5. Add deionized water dropwise to the obtained filtrate C at 60°C until turbidity appears, and then slowly cool it down to 5°C at a cooling rate of 0.5°C / min to crystallize. After filtration, collect the crystals and dry them in a vacuum oven at 50°C and 2.0 kPa to obtain about 68.7 g of high-purity deoxycholic acid.

[0032] The high-purity deoxycholic acid obtained in Example 1 above is pure white in color, with a purity of 99.4%, a specific rotation of +54.6, a melting point of 173-173.9℃, an arsenic content of about 0.00005%, and a heavy metal content of about 0.0003%.

[0033] Example 2

[0034] This embodiment 2 provides a method for preparing high-purity deoxycholic acid, which includes the following specific steps:

[0035] S1. Add 1000 ml of deionized water to a reaction apparatus equipped with a stirrer and a temperature controller. Then, under stirring conditions, add 100.0 g of industrial-grade deoxycholic acid (light pink, purity 95.5%, specific rotation +52.4, melting point 166.9~171.1℃, arsenic content 0.0003%, heavy metal content 0.002%). Continue to add 10% sodium hydroxide solution dropwise to the reaction apparatus until the pH of the system is 8~9 to obtain solution A.

[0036] S2. Add 700 ml of ethyl acetate to the above solution A for extraction. After stirring and mixing, separate the liquid and repeat the extraction and separation once to remove lipid-soluble impurities and macromolecular organic impurities in deoxycholic acid. Then, add 0.1 g of sodium sulfide (removing agent) and 1.0 g of pre-activated activated carbon (adsorbent) to the aqueous phase after extraction and stir at 50 °C for 1 hour to remove heavy metal ions and small molecule pigments respectively. Then, filter with medium-speed filter paper to remove activated carbon to obtain filtrate A.

[0037] S3. The above filtrate A is filtered a second time using a mixed fiber resin microporous filter membrane with a pore size of 0.45μm to obtain filtrate B. Then, hydrochloric acid is added dropwise to filtrate B to adjust the pH of filtrate B to 1-2, and the filter cake is collected after filtration.

[0038] S4. Add 500 ml of ethanol to the obtained filter cake and heat to 55°C. After complete dissolution, pass the hot solution through a PTFE microfiltration membrane with a pore size of 0.45 μm to obtain filtrate C.

[0039] S5. Deionized water was added dropwise to the obtained filtrate C at 55°C until turbidity appeared. Then, the temperature was slowly reduced to 6°C at a cooling rate of 1.0°C / min to crystallize. The filtrate was then filtered, the crystals were collected, and the crystals were dried in a vacuum oven at 55°C and 1.0 kPa to obtain approximately 69.3 g of high-purity deoxycholic acid.

[0040] The high-purity deoxycholic acid obtained in Example 2 above is pure white in color, with a purity of 99.2%, a specific rotation of +54.3, a melting point of 173-174°C, an arsenic content of approximately 0.00005%, and a heavy metal content of approximately 0.0004%.

[0041] Example 3

[0042] This embodiment 3 provides a method for preparing high-purity deoxycholic acid, which includes the following specific steps:

[0043] S1. Add 1800 ml of deionized water to a reaction apparatus equipped with a stirrer and a temperature controller. Then, under stirring conditions, add 100.0 g of industrial-grade deoxycholic acid (light pink, purity 95.5%, specific rotation +52.4, melting point 166.9~171.1℃, arsenic content 0.0003%, heavy metal content 0.002%). Continue to add 10% sodium hydroxide solution dropwise to the reaction apparatus until the pH of the system is 8~9 to obtain solution A.

[0044] S2. Add 400 ml of dichloromethane to the above solution A for extraction. After stirring and mixing, separate the liquid and repeat the extraction and separation once to remove lipid-soluble impurities and large molecular organic impurities in deoxycholic acid. Then, add 0.18 g of sodium sulfide (removing agent) and 5.0 g of pre-activated activated carbon (adsorbent) to the aqueous phase after extraction and stir at 55°C for 1.5 hours to remove heavy metal ions and small molecule pigments respectively. Then, filter with medium-speed filter paper to remove activated carbon to obtain filtrate A.

[0045] S3. The above filtrate A is filtered a second time using a mixed fiber resin microporous filter membrane with a pore size of 0.45μm to obtain filtrate B. Then, hydrochloric acid is added dropwise to filtrate B to adjust the pH of filtrate B to 1-2, and the filter cake is collected after filtration.

[0046] S4. Add 600 ml of ethanol to the obtained filter cake and heat to 60°C. After complete dissolution, pass the hot solution through a PTFE microfiltration membrane with a pore size of 0.45 μm to obtain filtrate C.

[0047] S5. Add deionized water dropwise to the obtained filtrate C at 60°C until turbidity appears, and then slowly cool it down to 5°C at a cooling rate of 0.5°C / min to crystallize. Then filter, collect the crystals, and dry the crystals in a vacuum oven at 50°C and 0.5KPa to obtain about 68.5g of high-purity deoxycholic acid.

[0048] The high-purity deoxycholic acid obtained in Example 3 above is pure white in color, with a purity of 99.1%, a specific rotation of +54.1, a melting point of 173-174°C, an arsenic content of approximately 0.00006%, and a heavy metal content of approximately 0.00045%.

[0049] It can be seen that the high-purity deoxycholic acid prepared by the method of the present invention has the advantages of extremely low arsenic and heavy metal content and high product purity (purity ≥ 99%), and can be used to process industrial-grade deoxycholic acid.

[0050] The above-described preferred embodiments of the present invention are for illustrative purposes only and are not intended to limit the scope of the invention. Any obvious variations or modifications derived from the technical solutions of the present invention are still within the protection scope of the present invention.

Claims

1. A method for preparing high-purity deoxycholic acid, characterized in that, The method includes the following steps: S1. Using industrial-grade deoxycholic acid as raw material, dissolve it in water and adjust the pH of the system to be alkaline to obtain solution A; S2. Add an extractant to the solution A for extraction and separation to remove lipid-soluble impurities and macromolecular organic impurities. Then add a desiccant and an adsorbent to the aqueous phase after extraction to remove heavy metal ions and small molecule pigments, respectively. Then filter to remove the adsorbent to obtain filtrate A. S3. Filter the filtrate A a second time to obtain filtrate B, then adjust the pH of filtrate B to acidic, filter it again, and collect the filter cake. S4. Add an organic solvent to the filter cake and heat until completely dissolved, then filter while hot to obtain filtrate C; S5. Add water dropwise to the filtrate C until it becomes turbid, then cool it to crystallize, continue filtering, collect the crystals, and vacuum dry to obtain high-purity deoxycholic acid.

2. The method for preparing high-purity deoxycholic acid according to claim 1, characterized in that, In step S1, the purity of deoxycholic acid is 90-97%; the mass-to-volume ratio of deoxycholic acid to water is 0.05-0.1 g / ml; and the pH of the system is adjusted to 8-9 using sodium hydroxide solution.

3. The method for preparing high-purity deoxycholic acid according to claim 1, characterized in that, The extractant in step S2 is either dichloromethane or ethyl acetate; the volume ratio of the extractant to water is 1:(1-5).

4. The method for preparing high-purity deoxycholic acid according to claim 1, characterized in that, The removal agent is sodium sulfide, and the adsorbent is activated carbon.

5. A method for preparing high-purity deoxycholic acid according to claim 1 or 4, characterized in that, The amount of the removal agent added is 0.01 to 0.2 wt% of the mass of industrial-grade deoxycholic acid, and the amount of the adsorbent added is 0.5 to 5.0% of the mass of industrial-grade deoxycholic acid.

6. The method for preparing high-purity deoxycholic acid according to claim 5, characterized in that, The amount of the removal agent added is 0.05 to 0.1 wt% of the mass of industrial-grade deoxycholic acid; the amount of the adsorbent added is 1.0 to 3.0% of the mass of industrial-grade deoxycholic acid.

7. The method for preparing high-purity deoxycholic acid according to claim 1, characterized in that, Step S3: The filtrate A is filtered a second time using a mixed fiber resin microporous membrane with a pore size of 0.45 μm to obtain filtrate B. Then, the pH of the filtrate B is adjusted to 1-2 with hydrochloric acid, filtered again, and the filter cake is collected.

8. The method for preparing high-purity deoxycholic acid according to claim 1, characterized in that, Step S4: Add ethanol to the filter cake and heat to 55-65°C. After complete dissolution, pass the hot solution through a PTFE microfiltration membrane with a pore size of 0.45 μm to obtain filtrate C.

9. The method for preparing high-purity deoxycholic acid according to claim 1, characterized in that, Step S5: Add deionized water dropwise to the filtrate C until turbidity appears, then slowly cool it to 3-7℃ at a cooling rate of 0.5-1.0℃ / min to crystallize, then filter, collect the crystals, and dry them in a vacuum oven at 45-60℃ and 0.5-2.0KPa to obtain high-purity deoxycholic acid.

10. A high-purity deoxycholic acid, characterized in that, The high-purity deoxycholic acid obtained by the preparation method according to any one of claims 1 to 9 has a purity ≥99%, a specific rotation of +54.0 to +57.0, a melting point of 173 to 174°C, an arsenic content <0.0001%, and a heavy metal content <0.0005%.