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Application of germacrone in preparation of medicament for treating or preventing influenza viruses

A technology for influenza A virus and influenza B virus, which is applied in the field of medicine and achieves the effect of being convenient to use

Inactive Publication Date: 2013-10-09
陈绪林
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no application of germacrone (Germacrone) in anti-influenza virus in current literature reports

Method used

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  • Application of germacrone in preparation of medicament for treating or preventing influenza viruses
  • Application of germacrone in preparation of medicament for treating or preventing influenza viruses
  • Application of germacrone in preparation of medicament for treating or preventing influenza viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A kind of application of gemmaconone in the preparation of treatment or prevention type A or type B influenza virus medicine, its steps are:

[0044] 1. Experimental materials and methods

[0045] 1.1 Cells, viruses and drugs

[0046] MDCK cells were purchased from ATCC, virus A / PR8 / 34H1N1 was amplified from chicken embryo culture, gemmacone was purchased from Sichuan Weikeqi Biotechnology Co., Ltd., and ribavirin was purchased from Hubei Tianyao Pharmaceutical Co., Ltd.;

[0047] 1.2 Experimental Instruments

[0048] Multifunctional detector PerkinElmer, inverted microscope Costar

[0049] 2. Experimental method:

[0050] 2.1 Cell culture:

[0051] Culture in a humidified incubator at 37° C., 5% (volume ratio, the same below) CO2. A DMEM medium containing 10% (volume ratio, hereinafter the same) FBS, 100 U / mL of penicillin and streptomycin was used. Cells are subcultured after reaching 90% confluence, and the subculture ratio is 1 / 3-1 / 4.

[0052] 2.2 Virus cultu...

Embodiment 2

[0095] Example 2: Inhibitory effect of gemmacone on different stages of virus replication cycle

[0096] 1. Cells are divided into 5*10 4 Inoculate in 24-well cell culture plates, and wait for the cells to adhere to the wall and form a monolayer for later use. The life cycle of influenza virus particles is about 6 hours, so we cultured with different drugs at the same concentration at 2-0 hours, 0-2 hours, 2-4 hours, and 4-6 hours after virus infection, and fixed the cells 6 hours after infection for indirect immunofluorescence. Detect the expression of viral NP protein, see the results Figure 7 .

[0097] 2. Experimental results show that gemmacone can inhibit virus infection when administered within -2~4h, but the effect is better when administered within -2~2h than 2~4h after infection, presumably mainly inhibiting early events of viral infection such as virus adsorption and entry . Ribavirin mainly inhibits inosine dehydrogenase, blocks the conversion of inosine-phosp...

Embodiment 3

[0099] Example 3: Research on broad-spectrum anti-influenza virus activity of gemmaconone

[0100] In this experiment, the antiviral activity of gemmaconone against influenza A virus subtype H3N2 strains, influenza A virus amantadine-resistant strains and type B influenza virus strains was detected.

[0101] 1. The virus strains used in the experiment include: A / human / Hubei / 3 / 2005(H3N2), A / WS / 33 / S31N(H1N1), Influenza B virus

[0102] 2. Divide MDCK cells into 2*10 4 Cells / well were seeded in 96-well cell culture plates, cultured in a 37°C cell culture incubator for 14-18 hours, and the cells were grown into a single layer before use. Discard the culture medium in the well plate, wash it twice with PBS solution, add 100TCID50 / well virus solution to infect the cells, and add a total of 200 μl of maintenance culture solution (DMEM+0.3%BSA+2.5ug / L trypsin) at the same time. After culturing in a cell incubator at 37°C for 48 hours, the supernatant of each experimental well was ta...

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Abstract

The invention discloses application of germacrone in preparation of a medicament for treating or preventing influenza viruses. The cell toxicity of the germacrone on madin-darby canine kidney (MDCK) cells is detected, and the result shows that CC50 is more than 250 microns. The influence of the germacrone on the expression of three antigens and the influence of germacrone on virus genome reproduction are detected, and the result shows that the germacrone formulation inhibits the expression of the virus antigens and the virus genome reproduction with dependency. The virus activity resistant mechanism of the germacrone is preliminarily researched. The virus activity resistance stage of the germacrone in a single reproduction period of the influenza viruses is researched by administration experiments of different time, and the result shows that the germacrone has inhibiting effect on adsorption, entry and early reproduction of the viruses. Moreover, the virus activity resistance of the germacrone to amantadine medicament resistant strains, B-type influenza virus strains and A-type influenza virus H3N2 subtype strains is detected, and the result shows that the germacrone has virus activity resistance to the three kinds of virus strains. The germacrone has the effects of relieving cough, relieving asthma and the like.

Description

technical field [0001] The invention belongs to the technical field of medicine, and more specifically relates to the application of gemmaconone in the preparation of medicines for treating or preventing influenza virus. Background technique [0002] Influenza virus is a pathogen that seriously affects human health and can cause influenza. According to the differences in the antigenic and genetic characteristics of the virion nucleoprotein (NP) and matrix protein (M), influenza viruses are divided into three types: A, B, and C, also known as A, B, and C. There are significant differences in the genome structure, polypeptide composition, infectivity and pathogenicity of these three different types of viruses. The whole genome of influenza A virus consists of eight single-stranded negative-strand RNAs of different sizes, named after segment 1 to segment 8, respectively. The total length of the viral genome is about 13.6kb, encoding 10 structural proteins (PB2, PB1, PA, HA, N...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/122A61P31/16
Inventor 陈绪林钱正旭
Owner 陈绪林
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