Method for inhibiting influenza virus infection and medicament thereof

A technology for influenza virus infection and drugs, applied in the field of treatment of enveloped virus infection, highly pathogenic avian influenza virus and human influenza virus, polypeptide and protein drugs, and treatment of influenza virus, which can solve the problem of interfering with the virus adsorption process, not influenza virus Specific inhibitors, drug-resistant strains, etc.

Inactive Publication Date: 2011-09-14
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] Embodiment 1: Construction of viral RNA expression plasmid of highly pathogenic avian influenza virus A / bar-headed goose / Qinghai / 1 / 05 strain HA and NA gene

[0056] 1.1) Extraction of viral RNA

[0057] Highly pathogenic avian influenza virus A / bar-headed goose / Qinghai / 1 / 05 strain (hereinafter referred to as QH strain) is a highly pathogenic avian influenza virus of H5N1 subtype, which was isolated and preserved by our laboratory (see literature Liu et al. al, 2005). Centrifuge the chicken embryo allantoic fluid of QH strain influenza virus stored at -70°C at 3000r / m for 10min to remove foreign proteins, then pipette 140μl of the supernatant into a RNase-free 1.5ml centrifuge tube, and use the viral RNA extraction kit (QIAmp Viral RNA Mini Kit, CAT.No.52904) was used to extract viral RNA, and the operation was performed according to the instructions provided by the kit. After extraction, adsorption, washing, centrifugation, and elution, 60 μl of viral RNA solution was...

Embodiment 2

[0073] Embodiment 2: the 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 (being H1N1 subtype, hereinafter referred to as "WSN") used in the experiment was prepared by our laboratory using the influenza virus reverse genetic system (Luytjes et al, 1989) with 12 plasmids. This plasmid reverse genetics system was donated by Professor Yoshihiro Kawaoka from the University of Wisconsin (Neumann et al, 1999). The system includes expression plasmids for expressing eight negative-strand RNA fragments of the 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 Escherichia coli DH5α competent cells, and the single clone...

Embodiment 3

[0078] Embodiment 3: Plaque titer determination of experimental influenza virus

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

[0080] Since dog kidney passage cells (MDCK) cells are s...

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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 therein. 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 poses 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 has important potential application value...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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