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Method for inhibiting influenza virus infection and medicament thereof

A technology for influenza virus infection and drugs, applied in the direction of viral peptides, antiviral agents, viruses/phages, etc., can solve the problems of interfering with the virus adsorption process, drug-resistant strains, and hindering the binding of HA to host cell membrane surface receptors

Inactive Publication Date: 2008-05-28
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(3) Artificially synthesized sialooligosaccharide analogues, such as sialoglycoside liposomes, sialoglycoside polymers, and divalent sialoglycoside analogs, can competitively bind HA, thereby hindering HA and host cell membranes The binding of surface receptors interferes with the adsorption process of the virus, thereby inhibiting the virus. At present, in vitro experiments have confirmed its inhibitory effect (Pang Haolong et al., 2004), but there are no reports of in vivo experiments, let alone commercialization The drug comes out
(4) At present, a variety of single and compound Chinese medicine preparations against influenza A virus have been screened out, mainly anti-inflammatory drugs and heat-clearing and detoxifying drugs (such as Zhang Weimin et al., 2001), which are used for supportive therapy, so they are not specific to influenza virus specific inhibitor
[0012] The above shows that although some progress has been made in the research and development of anti-influenza virus drugs, drug-resistant strains have appeared in commercialized chemically synthesized drugs. Among the anti-influenza drugs that have obtained patent protection, there is currently no commercialized, biotechnology-based Anti-influenza drugs developed

Method used

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  • Method for inhibiting influenza virus infection and medicament thereof
  • Method for inhibiting influenza virus infection and medicament thereof
  • Method for inhibiting influenza virus infection and medicament thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Construction of viral RNA expression plasmid of HA and NA genes of highly pathogenic avian influenza virus A / bar-headed goose / Qinghai / 1 / 05 strain

[0056] 1.1) Extraction of viral RNA

[0057] The highly pathogenic avian influenza virus A / bar-headed goose / Qinghai / 1 / 05 strain (hereinafter referred to as the QH strain) is a highly pathogenic avian influenza virus of the H5N1 subtype, which was isolated and preserved by our laboratory (refer to Liu et al. al, 2005). The QH strain influenza virus chicken embryo allantoic fluid stored at -70°C was centrifuged at 3000r / m for 10min to remove impurities, and then 140μl of supernatant was pipetted into a 1.5ml centrifuge tube without RNase, and viral RNA extraction kit (QIAmp Viral RNA Mini Kit, CAT. No. 52904) extract viral RNA and operate according to the instructions provided by the kit. After extraction, adsorption, washing, centrifugation, and elution, 60μl of viral RNA solution was obtained, which was stored at -80°...

Embodiment 2

[0073] Example 2: Preparation of recombinant influenza virus WSN and QH-WSN

[0074] 2.1) Preparation of recombinant influenza virus WSN

[0075] The recombinant human influenza virus A / WSN / 33 (H1N1 subtype, sometimes referred to as "WSN") used in the experiment was prepared by our laboratory using a 12-plasmid influenza virus reverse genetic system (Luytjeset al, 1989). The 12 This plasmid reverse genetic system was a gift from Professor Yoshihiro Kawaoka from the University of Wisconsin (Neumann et al, 1999). The system includes expression plasmids for expressing 8 negative-strand RNA fragments of influenza virus genome (including pHH21-PB2, pHH21-PB1, pHH21-PA, pHH21-HA, pHH21-NP, pHH21-NA, pHH21-M, pHH21 -NS,) and four protein expression plasmids expressing influenza virus replicase complex (including pcDNA3-PB2, pcDNA3-PB1, pcDNA3-PA, pCAGGS-NP). The above plasmids were transformed into competent cells of Escherichia coli DH5α, and the single clones were picked and cultured in ...

Embodiment 3

[0078] Example 3: Determination of plaque titer of experimental influenza virus

[0079] In order to test the inhibitory effect of the polypeptide and protein of the present invention on the highly pathogenic avian influenza virus, we tested its inhibitory effect on the QH-WSN recombinant virus with the H5N1 highly pathogenic avian influenza virus infection characteristics. At the same time, in order to test whether the above-mentioned peptides and proteins have cross-inhibition effects on human influenza viruses, we tested their inhibitory effects on recombinant WSN viruses (human influenza viruses) with similar genetic background as QH-WSN but with a surface membrane protein of H1N1 subtype. ; In addition, we also tested its cross-inhibitory effect (blue rain) against the representative strain of human influenza virus H3N2 subtype isolated from southern China in 2005 (A / Jiangxi / 312 / 2005, referred to as "JX" strain) Et al., 2006).

[0080] Since the dog kidney passage cell (MDCK)...

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Abstract

The invention relates to a method for restraining enveloped virus infection, relative polypeptide and protein drug, belonging to biological medicine technical field. The invention comprises a method for restraining influenza virus, particularly for restraining highly pathogenic influenza virus and human influenza virus (as H1N1 hypotype and H3N2 hypotype), relative polypeptide and protein, relative nucleic acid encoding the polypeptide and protein, and relative carriers and cells for representing the polypeptide and protein.

Description

Technical field [0001] The invention relates to a method for treating enveloped virus infection in the technical field of biomedicine and polypeptide and protein drugs used in the method. More particularly, the present invention includes methods for treating influenza viruses, especially highly pathogenic avian influenza viruses (such as H5N1 subtypes) and human influenza viruses (such as H1N1 subtypes and H3N2 subtypes), and the polypeptides and proteins involved therein. And the nucleic acid encoding the polypeptide or protein and the vector or cell capable of expressing the polypeptide or protein. Background technique [0002] Virus infection not only causes great harm to the health of all human beings, but also poses a serious threat to the survival and breeding of various animals, so it has become an important research topic in current medicine and related fields. The research and invention of drugs for the treatment of viral infections have important potential application v...

Claims

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Application Information

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IPC IPC(8): C07K14/11C07K14/00A61K38/16A61K38/08A61K38/10A61K48/00C12N7/00C12N15/63C12N15/44A61P31/16
Inventor 高福刘俊娥田波
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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