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Application of kavalactone compound in preparing medicine for inhibiting infection of enterovirus 71

A technology of kavasourin and enterovirus, which is applied in antiviral agents, pharmaceutical formulas, medical preparations containing active ingredients, etc.

Inactive Publication Date: 2017-12-01
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Enterovirus 71 infection has no effective treatment so far. For severe diseases such as myocarditis, encephalitis, and meningitis caused by virus infection, symptomatic treatment is mainly
There is no report about the use of kavasourin in the preparation of enterovirus 71 infection drugs

Method used

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  • Application of kavalactone compound in preparing medicine for inhibiting infection of enterovirus 71
  • Application of kavalactone compound in preparing medicine for inhibiting infection of enterovirus 71
  • Application of kavalactone compound in preparing medicine for inhibiting infection of enterovirus 71

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Screening of small molecule compounds for high-throughput inhibition of EV-A71 virus-infected cells

[0021] 1. Cells, viruses and compounds

[0022] RD (human rhabdomyomas) cells, Vero (African green monkey kidney) cells and BHK-21 (baby hamster kidney) cells in DMEM medium containing 1% penicillin / streptomycin and 10% fetal bovine serum at 37℃ , Cultivate in a 5% carbon dioxide incubator. The EV-A71 FY573 strain (GenBank accession number HM064456) is used for high-throughput drug screening, and the EV-A71 G082 strain is used for virus plaque reduction experiments and dosing experiments at different time points. EV-A71 virus strain SH12-276 (GenBank accession number KC570453), EV-A71 virus strain SH12-036 (GenBank accession number KC570452), CVA16 SHZH05-1 strain (GenBank accession number EU262658), Coxsackie virus B3 (CVB3, Nancy strain, ATCC VR-30) and type I poliovirus (PV1, Sabin strain) were used in the study of the antiviral spectrum of the compound. The...

Embodiment 2

[0025] Example 2: Determination of the inhibitory effect and cytotoxicity of kavapirin compounds on viruses

[0026] 1. Perform a virus infection inhibition test on different concentrations of kavakyorin

[0027] RD cells in the logarithmic growth phase are divided by 10 4 Cells / well were added to a 96-well cell culture plate and cultured for 24 hours. RD cells were infected with EV-A71 with a multiplicity of infection of 0.1, and a 3-fold dilution of kavakyorin was added. After 42 hours of culture, the supernatant was collected , Use plaque formation test to determine virus titer.

[0028] 2. Perform cytotoxicity experiments on different concentrations of kavakyorin

[0029] The experimental method is the same as the dose-dependent experiment, but no virus liquid is added. After culturing for 42 hours, 50 μl CellTiter-Glo reagent (Promega) was added, and the cell viability was judged by detecting the chemiluminescence intensity.

[0030] The results showed that kavatamycin can effect...

Embodiment 3

[0036] Example 3: Determination of virus titer

[0037] To measure the titer of EV-A71, add 1 ml of DMEM containing 3×105 RD cells to each well of a 12-well plate and culture for 24 hours. The virus was diluted by a factor of 10, that is, 25 μl of virus solution was mixed with 225 μl of DMEM containing 2% FBS and 1% penicillin / streptomycin. Aspirate the medium in the 12-well plate and add 200 μl of virus to each well. Place the infection in a 37°C, 5% carbon dioxide incubator for 1 h, and gently shake it every 15 minutes. Then aspirate the virus solution, add 1 ml of DMEM containing 0.8% methylcellulose and 2% fetal bovine serum, incubate for 6 days in a 37°C, 5% carbon dioxide incubator, and place it in 3.7% formalin for 1 h After that, stain with 1% crystal violet. For other viruses, CV-B3 was cultured for 1 day, PV1 was cultured for 2 days, and CV-A16 was cultured for 3 days before staining. The titer of EV-D68 is determined by half the tissue culture infectious dose (TCID...

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Abstract

The invention relates to application of kavalactone in preparing medicine for inhibiting infection of enterovirus 71 (EV-A71). Through cytology and molecular biology experiments, it is shown that the compound can specifically act on the VP4 protein of the EV-A71virus and can specifically inhibit the EV-A71 virus to enter a host cell; and through cytotoxicity tests, it is shown that the compound almost has no cytotoxicity. Therefore the compound can safely be used for preparing medicines for treating and preventing related diseases caused by infection of the enterovirus EV-A71.

Description

Technical field [0001] The present invention relates to the application of kavakyorein, in particular to the application of kavakyorein in preparing drugs for inhibiting enterovirus 71 infection. Background technique [0002] Enterovirus 71 (enterovirus 71, EV-A71) is a member of the Picornaviridae family (Picornaviridae), enterovirus genus (Enterovirus), and Coxsckie virus A16 (Coxsckie A16, CV-A16) is the cause of the Asia-Pacific region The main pathogen of hand, foot and mouth disease in infants and young children. According to statistics, from May 2008, China included hand, foot and mouth disease under the management of Category C infectious diseases to June 2015, a total of about 12.8 million cases of hand, foot and mouth disease were reported, of which more than 90,000 were severe cases and nearly 3,300 deaths. example. Currently, China ranks first in the world in both the number of morbidities and deaths from hand, foot and mouth disease. In 2014 alone, more than 2.7 m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/366A61P31/14
CPCA61K31/366
Inventor 黄海李桂明邹罡王莉莉
Owner SHANGHAI UNIV
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