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Preparation method for hydroxylation of 11 alpha of important intermediate of steroidal hormone substance

A technology of steroid hormones and intermediates, which is applied in the field of microbial transformation of steroid drugs, can solve the problems of low conversion rate and unfriendly environment, and achieve the effects of high conversion efficiency, improved utilization rate, and shortened production cycle.

Inactive Publication Date: 2014-08-06
HEBEI ZHONGSHENG BIOTECH
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  • Abstract
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Problems solved by technology

[0004] The present invention proposes a method for preparing 11α hydroxylation of important intermediates of steroid hormones, which is used to solve the problems of low conversion rate and unfriendly environment of steroid C11α hydroxylation in the existing microbial transformation

Method used

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  • Preparation method for hydroxylation of 11 alpha of important intermediate of steroidal hormone substance

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[0030] Such as figure 1 Shown: a preparation method of 11α hydroxylation of an important intermediate of steroid hormones, comprising the following steps:

[0031] The first step, strain propagation:

[0032] Insert the Ochraus ochrax or Nigerizomyces species into the corresponding seed medium for cultivation;

[0033] The second step, substrate emulsification:

[0034] Emulsifying one of the substrates 17α-hydroxyprogesterone, 4-androstenedione or 16,17-αepoxyprogesterone under the action of a surfactant;

[0035] The third step, fermentation culture:

[0036] Put the ochrax or nigeriasis obtained in the first step into the corresponding fermentation medium for a period of time, and then add sterilized 17α-hydroxyprogesterone, 4-androstenedione or 16,17- The emulsion of one of the α-epoxy progesterones continues to be fermented and cultivated;

[0037] The fourth step is to extract the finished product.

[0038] As a preferred technical solution, in another embodiment o...

Embodiment 1

[0053] A method for preparing an important intermediate of steroid hormones by 11α hydroxylation, comprising the following steps:

[0054] The first step, strain propagation:

[0055] Prepare 7 1L eggplant bottle slants of ochrax as a spore suspension and connect them in the seed medium of the seed tank for cultivation, wherein the amount of spores on each eggplant bottle slant is 10 10 -10 12 One, the seed medium includes the following concentrations of components: glucose 25-30g / L, corn steep liquor 20-30g / L, peptone 10-15g / L, yeast extract 5-10g / L, foam enemy 0.03%-0.04% , the pH of the seed medium is 3;

[0056] The second step, substrate emulsification:

[0057] The substrate 17α-hydroxyprogesterone is emulsified under the action of surfactant Tween 80 and temperature 50°C;

[0058] The third step, fermentation culture:

[0059] After 24 hours, transfer the seed culture solution obtained in the first step into the fermentation medium with an inoculation amount of 5% ...

Embodiment 2

[0066] Change the emulsification temperature in Example 1 to 60°C, and the rest of the operations are the same as in Example 1.

[0067] Test results: the conversion rate of 17α-hydroxyprogesterone is 98%, the final titer is 14.37g / L, and the purity of the obtained 11,17α-dihydroxyprogesterone is greater than 98.5%.

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Abstract

The invention provides a preparation method for hydroxylation of 11 alpha of an important intermediate of a steroidal hormone substance, and aims to solve the problems that the conversion rate is low and the environment is polluted when a microorganism is used for converting steroidal C11 alpha for hydroxylation in the prior art. The preparation method comprises the following steps: strain breeding, wherein a strain of ochratoxin or rhizopus nigricans is inoculated to a corresponding seed medium for cultivation; substrate emulsification, wherein a substrate selected from one of 17-alpha hydroxyl progesterone, 4-androstenedione or 16,17-alpha epoxy progesterone is subjected to emulsification treatment under the action of a surfactant; fermented cultivation, wherein the ochratoxin or rhizopus nigricans is inoculated to a fermented medium to be cultivated for a period of time, and then one of emulsion liquids of sterilized 17-alpha hydroxyl progesterone, 4-androstenedione or 16,17-alpha epoxy progesterone is added for performing continuous fermented cultivation; extracting a finished product. The method has the advantages of high conversion rate, little pollution, environmental protection, low pressure and the like.

Description

technical field [0001] The invention relates to the technical field of microbial transformation of steroid medicines, in particular to a preparation method for 11α hydroxylation of important intermediates of steroid hormones. Background technique [0002] Corticosteroids are a class of steroidal compounds synthesized and secreted by the adrenal cortex, and their main function is to regulate water-salt metabolism and glucose metabolism in animals. The C11α hydroxylation of steroids is an indispensable step in the synthesis of various corticosteroids. The hydroxyl groups on steroids are very difficult for chemical synthesis. Except for the C17 position of steroids, it is easy to introduce a hydroxyl group, and other positions are very difficult to introduce chemically. Steroidal C11α hydroxylation is a unique transformation reaction of microorganisms, which not only solves the problem of difficult drug synthesis, but more importantly, increases the activity of corticosteroids,...

Claims

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Application Information

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IPC IPC(8): C12P33/10C12R1/66C12R1/845
Inventor 宋浩雷张耀勋张敏然杨科
Owner HEBEI ZHONGSHENG BIOTECH
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