CN-116925173-B - Extraction method and application of triptolide

CN116925173BCN 116925173 BCN116925173 BCN 116925173BCN-116925173-B

Abstract

The application relates to the technical field of natural pharmaceutical chemistry, and particularly discloses a method for extracting triptolide and application thereof, wherein the method for extracting triptolide comprises the following steps of S1, preprocessing, namely cleaning and airing tripterygium wilfordii roots, and carrying out superfine grinding and plasma treatment to obtain tripterygium wilfordii powder; S2, performing enzymolysis on tripterygium wilfordii powder, inactivating enzyme, centrifuging, concentrating, drying to obtain a tripterygium wilfordii crude product, S3, purifying the tripterygium wilfordii crude product by purifying by a high-speed countercurrent chromatograph, and drying in vacuum to obtain a tripterygium wilfordii refined product, wherein the extraction method is simple and safe to operate, the purity and the extraction rate of the tripterygium wilfordii refined product are improved, and the prepared tripterygium wilfordii refined product can be mixed with total glucosides of paeony, glycyrrhizic acid, papain, poloxamer, xanthan gum and propylene glycol to prepare the medicament for treating rheumatoid arthritis, and has remarkable curative effect, high safety and wide clinical application prospect.

Inventors

JIN CHUNHUI
LV WENHUI
JIN LUMING
LIU BING

Assignees

浙江得恩德制药股份有限公司

Dates

Publication Date: 20260505
Application Date: 20230719

Claims (3)

1. The extraction method of triptolide is characterized by comprising the following steps of: s1, preprocessing, namely cleaning and airing the tripterygium wilfordii roots, and performing superfine grinding and plasma treatment to obtain tripterygium wilfordii powder; s2, performing enzymolysis, namely performing enzymolysis on the tripterygium wilfordii powder obtained in the step S1, inactivating enzyme, centrifuging, concentrating, and drying to obtain a tripterygium wilfordii crude product; s3, purifying the triptolide crude product obtained in the step S2 by a high-speed countercurrent chromatograph, and vacuum drying to obtain triptolide refined product; The enzymolysis comprises three steps of enzymolysis, wherein the first step of enzymolysis adopts cellulase and hemicellulase, the second step of enzymolysis adopts protease, amylase, xylanase and pectinase compound enzyme, and the third step of enzymolysis adopts beta-glucanase; the complex enzyme is prepared by mixing protease, amylase, xylanase and pectase in a mass ratio of 1:2:1:0.5; The plasma treatment condition is that the vacuum degree is 70-90Pa, the power is 600-800W, the treatment time is 1-2min, and the mixed gas consisting of nitrogen and oxygen with the volume ratio of 2:1 is introduced; The first step of enzymolysis is carried out at 50-55 ℃, the enzymolysis time is 3-6h, the pH is 4-5.5, the second step of enzymolysis is carried out at 45-50 ℃, the enzymolysis time is 8-10h, the pH is 4-6, the third step of enzymolysis is 48-60 ℃, the enzymolysis time is 4-8h, and the pH is 4.8-5.5; the solvent system of the high-speed countercurrent chromatograph is a mixed solution of 1-ethyl-3-methylimidazole acetate, normal hexane, chloroform, methanol and water, the volume ratio is 0.05:5-7:8-10:3-4:5, the lower phase is a mobile phase, and the upper phase is a stationary phase.
2. The method for extracting triptolide according to claim 1, wherein the mass ratio of the cellulase to the hemicellulase is 2:1-2.
3. The method for extracting triptolide according to claim 1, wherein the total mass of the cellulase and the hemicellulase is 1-2% of the tripterygium wilfordii powder, the mass of the compound enzyme is 2-4% of the tripterygium wilfordii powder, and the mass of the beta-glucanase is 0.5-1% of the tripterygium wilfordii powder.

Description

Extraction method and application of triptolide Technical Field The application relates to the technical field of natural pharmaceutical chemistry, in particular to an extraction method and application of triptolide. Background Triptolide is a natural compound extracted from tripterygium wilfordii plant, diterpenoid compound with complex chemical structure, white crystalline solid, which is almost insoluble in water at room temperature but soluble in organic solvent. Tripterine has various biological activities and pharmacological effects, on one hand, has strong anti-inflammatory and immunoregulatory effects, and can be used for treating autoimmune diseases and inflammatory diseases, and on the other hand, tripterine also has antitumor activity, and has inhibiting effect on various cancer cells including breast cancer, lung cancer, gastric cancer, liver cancer, etc. Thus, triptolide is widely studied in scientific research to explore its potential pharmacological effects and clinical applications. At present, CN105601700A discloses a method for preparing triptolide from tripterygium wilfordii, which comprises the steps of carrying out heating reflux extraction on tripterygium wilfordii roots by using an ethanol aqueous solution to obtain total extract, dissolving the total extract by using ethyl acetate for a plurality of times until the total extract is not dissolved any more to obtain an ethyl acetate part, sequentially carrying out chromatography on the ethyl acetate part by using a neutral alumina chromatographic column, a MCIGEL chromatographic column and a silica gel chromatographic column to obtain a tripterygium wilfordii ethyl extract crude product, and finally carrying out recrystallization by using ethyl acetate to obtain a tripterygium wilfordii ethyl extract pure product. Disclosure of Invention The application provides a method for extracting triptolide and application thereof, aiming at solving the problem of low extraction rate and purity of the existing triptolide. The extraction method of triptolide adopts the following technical scheme: A method for extracting triptolide comprises the following steps: s1, preprocessing, namely cleaning and airing the tripterygium wilfordii roots, and performing superfine grinding and plasma treatment to obtain tripterygium wilfordii powder; s2, performing enzymolysis, namely performing enzymolysis on the tripterygium wilfordii powder obtained in the step S1, inactivating enzyme, centrifuging, concentrating, and drying to obtain a tripterygium wilfordii crude product; s3, purifying the triptolide crude product obtained in the step S2 by a high-speed countercurrent chromatograph, and vacuum drying to obtain triptolide refined product; The enzymolysis comprises three steps, wherein the first step adopts cellulase and hemicellulase, the second step adopts protease, amylase, xylanase and pectinase compound enzyme, and the third step adopts beta-glucanase. According to the technical scheme, the tripterygium wilfordii root is crushed in the process of extracting tripterygium wilfordii ethyl, and then plasma treatment is carried out, so that a large number of tiny holes are formed on the surface of the tripterygium wilfordii powder, the contact area of the tripterygium wilfordii powder and enzyme is increased in the subsequent enzymolysis process, the enzymolysis efficiency can be improved, the release of the tripterygium wilfordii ethyl is accelerated, three-step enzymolysis is adopted in the enzymolysis process, the extraction rate of the tripterygium wilfordii ethyl is obviously improved, finally, the crude product is subjected to high-speed countercurrent purification, the purity of the tripterygium wilfordii ethyl is further improved, and the toxicity of the tripterygium wilfordii ethyl is greatly reduced. Preferably, the superfine grinding condition is that the rotating speed is 4000-6000rpm and the time is 20-30min. Preferably, the plasma treatment condition is that the vacuum degree is 70-90Pa, the power is 600-800W, the treatment time is 1-2min, and the mixed gas consisting of nitrogen and oxygen in the volume ratio of 2:1 is introduced. By adopting the technical scheme, the method adopts the plasma to treat the tripterygium wilfordii, on one hand, the method is favorable for destroying cell walls and cell membranes, changing surface properties and promoting the tripterygium wilfordii to be more easily dissolved or released in the tripterygium wilfordii, on the other hand, high-energy particles and active species generated by the plasma can destroy microorganisms such as bacteria, fungi and viruses, and the like, and remove microbial pollutants in the tripterygium wilfordii, so that the purity and quality of the tripterygium wilfordii are improved. Preferably, the mass ratio of the cellulase to the hemicellulase is 2:1-2. Preferably, the complex enzyme is prepared by mixing protease, amylase, xylanase and pectinase in a mass ratio of 1:2:1:0.5. Preferab