Separating purifying method for ingenol extract

Abstract

The invention relates to a separating purifying method for ingenol extract, and belongs to the field of separation of traditional Chinese medicine. The to-be-solved technical problem is to provide a separating purifying method for ingenol extract. The separating purifying method for ingenol extract comprises the following steps: a, adding an alkali into ingenol extract for hydrolysis, adding an acid for neutralizing, and concentrating to obtain a paste; b, separating and purifying the paste through silica gel column chromatography; and c, performing reduced-pressure concentration on the ingenol ethyl acetate solution collected in the step b, standing for precipitating, and filtering to obtain ingenol. The method is simple in operation, avoids multi-step extraction operation, the yield is improved, and also large-scale continuous production is realized. Silica gel in chromatography column B can be repeatedly used for two or more times, the silica gel usage amount and the reagent consumption quantity are reduced to a certain degree, and production cost is substantially reduced.

Description

technical field [0001] The invention relates to a separation and purification method of ingenol extract, belonging to the field of extraction and separation of traditional Chinese medicines. Background technique [0002] Ingenol is a natural product of tetracyclic diterpenoids. Its structural formula is shown in formula I. It is derived from Euphorbiaceae Euphorbia plants. Anti-HIV and other biological activities, among which Picato, a new drug with ingenol-3-angelate as the molecular entity, was approved by the FDA in 2012 for the treatment of actinic keratosis. [0003] [0004] The structure of ingenol is novel and complex. In the past 30 years, although its unique structure and activity have attracted the attention of many chemists, there are also reports on total synthesis. Increased market demand; LEOPharma company cooperates with Scripps Research Institute [Science341,878 (2013)], obtains ingenol by (+)-3-carene through 14 steps final synthesis, and total yield re...

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Problems solved by technology

[0006] Chinese patent CN102126941A uses extraction, back extraction, alkali hydrolysis, pH adjustment, and concentration under reduced pressure to extract the extract, and then uses silica gel column chromatography on the concentrated extract, combined with recrystallization means to purify to obtain high-purity ingenol. In the early stage of the method, there are multiple extraction operations, which are time-consuming, low transfer rate, large loss, and the expensive and toxic extraction reagent acetonitrile is used, which is high in cost, not conducive to environmental protection, and is not suitable for large-scale production, and the silica gel column layer The analysis adopts petroleum ether / ethyl acetate (1:1) to elute and remove impurities, and then add ethyl acetate to elute. In the process of removing impurities, part of the ingenol compound will be taken out due to the addition of ethyl acetate, reducing the Yield

[0007] Patent CN103694096A also adopts the extraction operation to extract the extract. After the extract is concentrated, it undergoes silica gel column chromatography and preparative liquid phase purification, and finally obtains high-purity ingenol monomer. It exists in the form of ester derivatives, and the purification process does not use a hydrolysis step, so the yield of the final product is extremely low, and it is difficult to meet the requirements of large-scale production

[0008] In the patent WO2013050365A1, the medicinal material powder of Stephania chinensis is directly hydrolyzed by adding alkali and stirring, after neutralization, it is extracted with petroleum ether and tetrahydrofuran in sequence, and then purified by silica gel column chromatography to obtain high-purity ingenol. Although this method simplifies the purification steps, but Because the medicinal powder is directly added with alkali and stirred for hydrolysis, there is no guarantee that the ingenol derivatives in the medicinal materials can be extracted and hydrolyzed completely

[0009] The above-mentioned patents all use multi-step extraction operations. Because the stephenia is rich in oil components, it is easy to emulsify during extraction, and multiple extractions with a large amount of solvent are required to completely extract the target components. The extraction efficiency is low, resulting in a low extraction rate; the extraction used for extraction Agent sherwood oil has a low boiling point, and the recovery rate of concentration under reduced pressure is low, which also increases production costs to a certain extent; Remove less polar impurities. Due to the addition of ethyl acetate in the mixed eluent, the target compound ingenol will also be taken out along with the impurities, reducing the production yield

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