Method for preparing dehydroepiandrosterone through chemical-enzyme method

A technology for the preparation of dehydroepiandrosterone and enzymatic method, which is applied in the field of chemical-enzymatic preparation of dehydroepiandrosterone, can solve the problems of difficult removal of impurities, high amount of organic solvent, high amount of potassium tert-butoxide, etc., and achieve high practicality The effect of high value, high product yield and high purity

Active Publication Date: 2016-11-09
ENZYMEWORKS
View PDF8 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the reported 4-AD to 5-AD reaction, the purity of 5-AD is the main problem affecting the next step of the reaction, because its impurities are difficult to remove, and there is generally a high amount of potassium tert-butoxide, and the product 5-AD has a high yield. The problem of low yield, such as the amount of potassium tert-butoxide in the patent WO2014188353 is 5 times the equivalent of ...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for preparing dehydroepiandrosterone through chemical-enzyme method
  • Method for preparing dehydroepiandrosterone through chemical-enzyme method
  • Method for preparing dehydroepiandrosterone through chemical-enzyme method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of 5-AD

[0024] At 25° C., nitrogen was bubbled into tert-butanol (80 mL) for 15 minutes, potassium tert-butoxide (6.0 g, 3.0 eq) was added under aeration conditions, and the reactor was sealed, replaced with nitrogen, and stirred for half an hour. Under the protection of nitrogen, the substrate (5.0g, 1.0eq) was added, and the reaction was carried out at 35°C for 5 hours. Take 3.5 g of sodium ascorbate, 5.7 g of acetic acid, and 200 mL of water, stir for 15 minutes, pour the above-mentioned reaction solution, and stir for half an hour under nitrogen protection. The conversion rate detected by HPLC is 96.5%. The mixed solution was filtered to obtain a white solid, washed with water, and spin-dried to obtain 4.8 g of a white solid with a purity of 95.5% by HPLC and a yield of 91.7%.

Embodiment 2

[0025] Example 2 Preparation of dehydroepiandrosterone

[0026] Dissolve the above 4.8g product in 25mL 2-methyltetrahydrofuran, add 25mL 100mM pH6.5 phosphate buffer solution to start stirring, add 5g glucose, 80mg magnesium chloride hexahydrate, 60mg ketoreductase (from sequence 26 in E. coli Obtained from medium expression), 30mg glucose dehydrogenase (from Suzhou Hanzyme Biotechnology Co., Ltd., numbered EW002), 20mg NAD, start the reaction at 25°C, and use 40% sodium hydroxide to control pH 6.5. After 6 hours, take a sample for detection. Conversion rate 99%, adjust pH with hydrochloric acid 99%, content> After 99% of the mother liquor is evaporated to dryness, 1.4 g of solid is obtained. After re-recycling, the multi-batch post-processing yield is 85%.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for preparing dehydroepiandrosterone through a chemical-enzyme method. The method comprises that dehydroepiandrosterone is prepared from 4-androstenedione as an initial substrate orderly through a chemical method and a biological method. In preparation, the two reaction processes are optimized. In the chemical method-based 5-androstenedione preparation process, a reaction solution is added into an aqueous solution with sodium ascorbate and acetic acid so that reaction conditions are mild. In the second biological method-based dehydroepiandrosterone preparation process, a ketoreductase is used as a catalyst so that the product yield and purity are improved. In the whole reaction processes, use amounts of a coenzyme and potassium tert-butoxide are low and a high practical value is obtained.

Description

Technical field [0001] The invention relates to the field of pharmacy, in particular to a method for preparing dehydroepiandrosterone by a chemical-enzymatic method. Background technique [0002] Dehydroepiandrosterone (DHEA) has the structure shown in Formula 1. It is a steroid compound secreted by the adrenal cortex of the human body. It has the effects of delaying aging and resisting diseases. It is also a precursor for the synthesis of other steroid drugs. [0003] [0004] Among the methods for preparing dehydroepiandrosterone that have been published, most of them are fermentation and chemical methods. The fermentation method of adding the precursor to the microbial fermentation broth for biotransformation is reported in the patent CN 201210316197, etc., which takes a long time, the product content is less than 1 g / L, and there are problems such as separation and extraction. Starting with other steroids, such as reported in patents CN102212099, 102603841, 102603839, etc., pr...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12P33/06C07J1/00
CPCC07J1/0011C12P33/06
Inventor 张跃
Owner ENZYMEWORKS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap