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Application of mepyramine maleate in preparing drug for treating or preventing influenza virus

A kind of pyramine maleate, influenza prevention technology, applied in the field of medicine, can solve the problem of no pyramine maleate application and the like

Inactive Publication Date: 2014-12-17
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no application of pyrilamine maleate against influenza virus

Method used

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  • Application of mepyramine maleate in preparing drug for treating or preventing influenza virus
  • Application of mepyramine maleate in preparing drug for treating or preventing influenza virus
  • Application of mepyramine maleate in preparing drug for treating or preventing influenza virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Evaluation of anti-influenza virus activity of pyrilamine maleate

[0032] 1. Experimental materials and methods

[0033] 1.1 Cells, viruses and drugs

[0034] MDCK cells were purchased from ATCC, virus A / PR8 / 34H1N1 was amplified from chicken embryo culture, and pyrilamine maleate was purchased from Sigma;

[0035] 1.2 Experimental Instruments

[0036] Multifunctional detector PerkinElmer, inverted microscope Costar

[0037] 2. experimental method:

[0038] 2.1 Cell culture:

[0039] 37°C, 5% (volume ratio, the same below) CO 2 cultured in a humidified incubator. A DMEM medium containing 10% (volume ratio, the same below) FBS, 100 U / mL of penicillin and streptomycin was used. Cells were subcultured after reaching 90% confluence, and the subculture ratio was 1 / 3–1 / 4.

[0040] 2.2 Virus culture:

[0041] Take 9-11 day-old SPF chicken embryos, check them with an egg tester before inoculating the virus, and mark them at a position far away from the ...

Embodiment 2

[0057] Example 2: Inhibitory Effects of Pyramine Maleate on Different Stages of the Viral Replication Cycle

[0058] 1. cells by 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 is about 6 hours, so we gave 50μM drug culture at virus infection -2~0h, 0~2h, 2~4h, 4~6h respectively, fixed the cells at 6h after infection, and performed indirect immunofluorescence according to the following steps The expression levels of viral proteins after drug treatment were detected.

[0059] 2. Discard the supernatant of the culture medium, fix the cells with fixative solution at room temperature for 15-20min; wash 3 times with PBS, 5min each time; monoclonal antibody) and incubate at room temperature for 1 h; wash with PBS 3 times, 5 min each time; add fluorescently labeled secondary antibody diluted in binding solution and DAPI staining solution and incubate at room tempera...

Embodiment 3

[0062] Example 3: Study on broad-spectrum anti-influenza virus activity of pyrilamine maleate

[0063] This experiment detects the antiviral activity of pyrilamine maleate against influenza A virus subtype H1N1 strain, influenza A virus H3N2 strain and influenza B virus strain.

[0064] 1. The virus strains used in the experiment include: A / human / Hubei / 1 / 2009 (H1N1), A / human / Hubei / 3 / 2005 (H3N2), Influenza B virus

[0065] 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 drugs of various concentration gradients at the same time (with 200 μ M as the initial concentration, serially dilute 6 gradients by 2 times, each gradient 2 A total of 200 μl of maintenance medium (DMEM+0.3%BSA+2.5...

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Abstract

The invention discloses application of mepyramine maleate in preparing a drug for treating or preventing influenza virus. The application comprises the following steps of: selecting the mepyramine maleate with complete non-toxic concentration for carrying out an antiviral experiment, wherein the result shows that the compound has remarkable antiviral activity and is dose-dependent; analyzing an anti-influenza virus action period of the mepyramine maleate, wherein the result indicates that the mepyramine maleate can be mainly used for inhibiting virus infection early events including virus adsorption, virus entry and the like; and detecting the antiviral activity of the mepyramine maleate to different types and subtypes of influenza viruses, wherein the result shows that the mepyramine maleate can be developed as a novel anti-influenza virus drug, so that a novel way and means is provided for treating the influenza.

Description

technical field [0001] The invention belongs to the technical field of medicine, and more specifically relates to an application of pyrilamine maleate 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,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/4402A61P31/16
Inventor 陈绪林廖庆姣
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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