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A kind of method utilizing sphingomonas to split hepatitis C ns3 enzyme inhibitor medicine intermediate

A technology of sphingomonas and hepatitis C, which is applied in the field of splitting hepatitis C NS3 enzyme inhibitor drug intermediates by using sphingomonas, which can solve the problems of low split efficiency and large usage , to achieve the effect of high resolution activity, shortened enzyme dosage and low price

Active Publication Date: 2019-08-02
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, there is no report on the use of microbial whole-cell method for splitting experiments. At the same time, the existing enzymatic splitting efficiency is low and the amount of use is large.

Method used

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  • A kind of method utilizing sphingomonas to split hepatitis C ns3 enzyme inhibitor medicine intermediate
  • A kind of method utilizing sphingomonas to split hepatitis C ns3 enzyme inhibitor medicine intermediate
  • A kind of method utilizing sphingomonas to split hepatitis C ns3 enzyme inhibitor medicine intermediate

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Embodiment 1

[0049] Example 1: Method for splitting N-Boc-1-amino-2-vinylcyclopropane ethyl ester and 1-amino-2-vinylcyclopropane ethyl ester hydrochloride in whole cells of the strain of the present invention

[0050] (1) Strain selection: use Sphingomonas aquatilis JSS7;

[0051] (2) Seed culture: Inoculate 100 μL of the stored bacterial solution into 50 mL of seed culture medium, and culture it at 30° C. and 200 rpm for 24-48 hours to fully grow the bacteria. Observe that the culture medium is obviously turbid in color.

[0052] (3) Collect the bacteria: centrifuge the bacteria at low temperature and discard the supernatant. The cells were suspended in 10 mL of phosphate buffer for use.

[0053] (4) Transformation experiment: The substrate concentration is still 1g / L, and the transformation experiment can be completed at 30°C and 200rpm for 72 hours.

[0054] (5) Detection: Extract the reaction solution with equal volume of n-hexane, take the organic phase after centrifugation, inject 10 μL of s...

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Abstract

The invention discloses a method for splitting a hepatitis c NS3 enzyme inhibitor drug intermediate through sphingomonasaquatilis, and particularly relates to a method for splitting hepatitis c NS3 enzyme inhibitor drug intermediate N-Boc-1-amino-2-vinylcyclopropane ethyl formate and 1-amino-2-vinylcyclopropane ethyl formate hydrochloride to prepare (1R, 2S)-N-Boc-1-amino-2-vinyl cyclopropane ethyl formate and (1S, 2R)-1-amino-2-vinyl cyclopropane ethyl formate. The method comprises the following steps that a strain is introduced into a culture medium with N-Boc-1-amino-2-vinylcyclopropane ethyl formate or 1-amino-2-vinylcyclopropane ethyl formate hydrochloride as a unique carbon source, a whole-cell catalysis substrate obtained through screening is subjected to a split reaction, and (1R, 2S)-N-Boc-1-amino-2-vinyl cyclopropane ethyl formate and (1S, 2R)-1-amino-2-vinyl cyclopropane ethyl formate are obtained.

Description

Technical field [0001] The invention relates to the use of a sphingosine to separate the hepatitis C NS3 enzyme inhibitor drug intermediate N-Boc-1-amino-2-vinylcyclopropane ethyl ester and 1-amino-2-vinyl Ethyl cyclopropane carboxylate hydrochloride to prepare (1R,2S)-N-Boc-1-amino-2-vinylcyclopropane ethyl carboxylate and (1S,2R)-1-amino-2-vinylcyclopropane Ethyl formate method. Background technique [0002] Hepatitis C virus (HCV) belongs to the Hepatitis C virus genus of the Flaviviridae family, and includes six genotypes, including I, II, and III. It has been more than 20 years since its discovery, and more than 170 million people have been infected worldwide. HCV virus includes structural proteins and non-structural proteins. Among them, the non-structural protein NS3 enzyme can directly destroy host cells and inhibit the response to interferon, which seriously affects the treatment of it. There is no safe and effective virus vaccine against HCV. Therefore, the research a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/00C12P41/00C12N1/02C12N1/20C12R1/01
CPCC12N1/02C12N1/20C12P13/00C12P41/00
Inventor 郑国钧石莹
Owner BEIJING UNIV OF CHEM TECH
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