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Method for screening SARS coronaviruses main Protease inhibitors from natural product such as Chinese medicinal materials mixture prepared storeroom

A coronavirus, main protease technology, applied in library screening, chemical library, combinatorial chemistry and other directions, to achieve the effect of high screening efficiency, low cost and simple method

Active Publication Date: 2007-10-31
NANKAI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is no technology to screen SARS-CoV main protease inhibitors based on the crystal structure of SARS-CoV main protease from natural medicine systems such as traditional Chinese medicine (such as natural product mixture candidate libraries such as Chinese medicine)

Method used

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  • Method for screening SARS coronaviruses main Protease inhibitors from natural product such as Chinese medicinal materials mixture prepared storeroom
  • Method for screening SARS coronaviruses main Protease inhibitors from natural product such as Chinese medicinal materials mixture prepared storeroom
  • Method for screening SARS coronaviruses main Protease inhibitors from natural product such as Chinese medicinal materials mixture prepared storeroom

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Experimental program
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Embodiment 1

[0056] 1. Expression, purification and crystal growth of SARS coronavirus main protease

[0057] According to the literature (Yang H etal.Proc Natl Acad Sci.2003 Nov; 100 (23): 13190-5), the expression, purification and crystal growth of the main protease of SARS coronavirus (see the sequence listing part for the amino acid sequence) were carried out to obtain the diffraction resolution Higher (higher than 2.6 Ȧ) crystals of SARS coronavirus main protease. specifically:

[0058] The expression vector construction of a.SARS coronavirus main protease

[0059] Specific steps include:

[0060] (1.1) Using the cDNA library of the SARS virus strain numbered BJ01 provided by Beijing Huada Genomics Center, PCR technology was used to amplify in vitro.

[0061] Forward primer: 5′-CGGGATCCAGTGGTTTTAGGAAAATG-3′

[0062] Reverse primer: 5′-CCGCTCGAGTCATTGGAAGGTAACACCAGA-3′

[0063] (1.2) After the gene fragment amplified by PCR was digested with BamHI and XhoI double enzymes, a fragme...

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Abstract

The invention discloses a method to sieve SARS coronavirus main prolease depressor from natural products mixture optional warehouse, which comprises the following steps: touching crystal of SARS coronavirus main prolease with 'sample with inhibited active'; getting electron density of small molecule depressant combined with SARS coronavirus main prolease; appraising the small molecule depressant based on electron density; getting fine three-dimensional structure of SARS coronavirus main prolease and small molecule depressant; getting small molecule depressant Sichuan scented sea vegetable ala factor. This Sichuan scented sea vegetable ala factor can inhibit coronavirus main prolease activator effectively, which can cure or prevent SARS coronavirus and infectivity gastrointestinal disease virus.

Description

technical field [0001] The present invention relates to the screening method of medicine, more specifically, relate to based on the crystal structure of SARS coronavirus main protease, the new method of screening the inhibitor of SARS coronavirus main protease from the natural product mixture such as traditional Chinese medicine mixture library and based on The inhibitor of the SARS coronavirus main protease discovered by this method. Background technique [0002] SARS (Severe Acute Respiratory Sydrome) is the abbreviation for causing severe acute respiratory syndrome (atypical pneumonia), and its pathogen is a virus of the genus Coronaviridae (Peiris J. et al. Lancet, 2003, 361: 1319-1325) . Coronaviruses are positive-strand RNA viruses. Coronaviruses belong to the family Coronaviridae. Among the currently known positive-strand RNA viruses, their genomes are the largest (Siddell, S.G. et al. Coronaviruses, toroviruses, and arteriviruses. in Top...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B30/04G01N33/573
Inventor 饶子和孙玉娜娄智勇向科辉
Owner NANKAI UNIV
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