Application of nucleotide-binding oligomerization domain-like receptor protein in the treatment of Zika virus infection

A technology of oligomerization domain and receptor protein, which can be applied to antiviral agents, peptide/protein components, medical preparations containing active ingredients, etc., and can solve the problems of Zika virus prevention and treatment.

Active Publication Date: 2020-06-30
广东龙帆生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are no reports on the preventive and therapeutic effects of NLRP3 itself on Zika virus (ZIKV)

Method used

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  • Application of nucleotide-binding oligomerization domain-like receptor protein in the treatment of Zika virus infection
  • Application of nucleotide-binding oligomerization domain-like receptor protein in the treatment of Zika virus infection
  • Application of nucleotide-binding oligomerization domain-like receptor protein in the treatment of Zika virus infection

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Experimental program
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Embodiment 1

[0031] The application of a nucleotide-binding oligomerization domain-like receptor protein 3 (NOD-like receptors, NLRP3) in the preparation of a drug for treating or preventing Zika virus (ZIKV) infection, the steps of which are:

[0032] (1) Clone NLRP3 gene (NLRP3 preparation): using the total mRNA extracted from THP-1 (Human acute monocytic leukemia cell line) cells as a template, amplify the total cDNA by reverse transcription PCR, and then use the total cDNA as a template The template was amplified with specific primers to obtain the NLRP3 gene fragment. Primers are as follows:

[0033] P1 (forward primer): 5'-CGCGGATCCATGAAGATGGCAAGCACCCGC-3'BamHI

[0034] P1 (reverse primer): 5'-CCGCTCGAGCTACCAAGAAGGCTCAAAGAC-3' XhoⅠ

[0035] The NLRP3 gene was cloned into the pTAC (purchased from Cresbo) promoter of the vector pGEX-6p-1 (purchased from YRgene) by BamHI and XhoI digestion and ligation to obtain the expression vector pGEX-6p-1-NLRP3 of NLRP3 .

[0036] (2) Acquisiti...

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Abstract

The invention discloses application of nucleotide binding and oligomerization domain-like receptor protein in the treatment of Zika virus infection drugs. The steps include: A, evaluating activity ofthe nucleotide binding and oligomerization domain-like receptor protein e to resist Zika virus; B, extracting total mRNA in THP-1 cells to form a template; performing reverse PCR (polymerase chain reaction) amplification to obtain total cDNA; performing Bam H I and Xho I enzymatic cutting and connection so as to clone NLRP3 gene to pTAC starter of the carrier pGEX-6p-1; C, selecting the single strain colony, introducing LB liquid culture with ampicillin, performing NLRP3 expression, purifying and concentrating by means of affinity chromatography to obtain NLRP3 lyophilized liquid; D, adding aprotein into the cells, and detecting ZIKV NS5 mRNA level and protein level; NLRP 3 is in-vivo composed inertial expression and has no side and effective actions for the human body; pathogenesis of the NLRP3 refers to inhibit virus copy by changing signals in cells, anti-virus mechanisms are diverse, and viruses and drug resistance are avoided.

Description

technical field [0001] The present invention relates to the field of antiviral drugs, more specifically to the use of a nucleotide-binding oligomerization domain-like receptor protein 3 (NOD-like receptors, NLRP3) in the preparation of drugs for treating or preventing Zika virus (ZIKV) infection application. Background technique [0002] Zika virus (ZIKV) is an arbovirus transmitted through the bite of Aedes mosquitoes. ZIKV was first discovered in 1947 in a febrile rhesus monkey in the Zika Forest of Uganda. Despite serological evidence that ZIKV is widespread worldwide, few human infections with ZIKV have been documented. Until 2007, a large-scale outbreak of Zika virus occurred on Yap Island, Federated States of Micronesia. 75% of the residents on this island in the South Pacific-like area were infected by the virus. This is the first time that ZIKV has shown its potential. Transmissibility and pathogenicity. The second large-scale outbreak of ZIKV occurred in French ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P31/14
CPCA61K38/177A61P31/14Y02A50/30
Inventor 吴建国王文标邬开朗刘映乐李耿刘芳谭秋萍罗震
Owner 广东龙帆生物科技有限公司
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