Novel use of cinobufagin and resibufogenin in inhibition of enterovirus type 71 infection

A technology of cinobufafonitoxin and esterbufagenin, which can be used in antiviral agents, medical preparations containing active ingredients, and pharmaceutical formulas, and can solve problems such as antiviral effects that have not been reported

Inactive Publication Date: 2016-05-11
JILIN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antiviral effects of cinobufagin and bufagenin on enterovirus 71 have not been reported yet.

Method used

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  • Novel use of cinobufagin and resibufogenin in inhibition of enterovirus type 71 infection
  • Novel use of cinobufagin and resibufogenin in inhibition of enterovirus type 71 infection
  • Novel use of cinobufagin and resibufogenin in inhibition of enterovirus type 71 infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Construction of RDS cells

[0035] The synthetic EV71 / CA16 receptor hSCARB2 gene fragment (NCBI accession number: NM_005506.3, synthesized by Shanghai Jierui Company) with EcoRI&XbaI restriction sites at both ends was inserted into the lentiviral vector Lenti-XpLVX-Puro vector (purchased from Clontech ), and use the transfection reagent in the lentiviral vector kit (purchased from Clontech) to transfect human rhabdomyosarcoma cells (RD cells, purchased from ATCC, accession number #CCL-136) according to the instructions to obtain RD stably transfected with hSCARB2 cells, referred to as RDS cells (for the specific construction method of RDS cells, refer to Lietal. Virology Journal, 10:250).

Embodiment 2

[0036] Example 2: Cytotoxicity of Cinobufagin and Esterbufagenin on Cells and Inhibition of Cytopathic Effects Caused by Enterovirus 71

[0037] Take RD cells and RDS cells in the logarithmic growth phase, and divide them into 3×10 4 pcs / hole and 6×10 4 The concentration of cells / well was inserted into a 96-well culture plate and cultured for 24 hours. Both RD cells and RDS cells were incubated at 37 °C and 5% CO 2 Under culture, wherein the culture medium used for cultivating RD cells is DMEM medium supplemented with 10% fetal bovine serum (referred to as 10% FBS-DMEM, FBS (fetal bovine serum) and DMEM are both purchased from Sigma), cultivating RDS cells The culture medium used was 10% FBS-DMEM containing 0.5 μg / ml puromycin (purchased from Clontech).

[0038] Add 50 μL of DMEM cell culture medium (purchased from Sigma) containing different concentrations of cinobufagin or esterbufagenin (purchased from China Institute for the Control of Pharmaceutical and Biological Prod...

Embodiment 3

[0060] Example 3: Inhibition of Cinobufagin and Esterbufagenin on Plaque Formation Caused by EV71

[0061] Take the RDS cells in the logarithmic growth phase, and use 6×10 5 The concentration of each well was inserted into a 12-well culture plate and cultured for 24 hours.

[0062] Aspirate the medium from each well, add 300 μL of DMEM cell culture medium containing different concentrations of cinobufacien to make the final concentration 62.50nM, 31.25nM, 15.63nM, 7.81nM, 3.90nM or add 300μL of cinobufacin containing different concentrations Base DMEM cell culture solution, so that the final concentration is 1563nM, 781nM, 391nM, 195nM, 98nM, each concentration is set up 3 duplicate wells; add 300μ LEV71 (100PFU / hole) virus infection to each hole at the same time of adding medicine; And set only Add the cell control of DMEM cell culture medium and the virus control of only adding EV71 infection; after continuing to culture the cells for 1 hour, aspirate the supernatant of th...

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Abstract

The invention relates to a novel use of cinobufagin and resibufogenin and concretely relates to a use of cinobufagin and resibufogenin in preparation of drugs for preventing and treating the hand-foot-and-mouth disease and especially in preparation of drugs for resisting enterovirus type 71.

Description

technical field [0001] This application relates to the new application of cinobufagin and bufagenin, specifically, the use of cinobufagin and bufagenin in the preparation of medicines for the prevention and treatment of hand, foot and mouth disease, especially the use of It is used in the preparation of anti-enterovirus 71 drugs. Background technique [0002] HFMD is a common acute viral infectious disease in children. The main pathogens are Enterovirus 71 (abbreviated as EV71) and Coxsackievirus A16 (Coxsackievirus A16, abbreviated as CA16). The disease is particularly serious. In recent years, frequent outbreaks of HFMD in the Asia-Pacific region have become a serious social health problem. According to statistics, from 2008 to 2012 in my country, more than seven million cases of hand, foot and mouth disease were reported, resulting in 2443 deaths. [0003] There is currently no commercially available vaccine to prevent HFMD. In terms of treatment, conventional antivir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/585A61P31/14
Inventor 苏维恒姜春来孔维陈嘉雯
Owner JILIN UNIV
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