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Application of diphenhydramine hydrochloride in preparation of medicines for treating or preventing influenza viruses

A technology of diphenhydramine hydrochloride and influenza virus, applied in the field of medicine, can solve the problem of no diphenhydramine hydrochloride against influenza virus and the like, and achieve the effect of broad-spectrum antiviral activity

Inactive Publication Date: 2014-11-12
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

Diphenhydramine hydrochloride can be used to treat rhinitis, currently there is no direct application of diphenhydramine hydrochloride in anti-influenza virus

Method used

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  • Application of diphenhydramine hydrochloride in preparation of medicines for treating or preventing influenza viruses
  • Application of diphenhydramine hydrochloride in preparation of medicines for treating or preventing influenza viruses
  • Application of diphenhydramine hydrochloride in preparation of medicines for treating or preventing influenza viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Evaluation of anti-influenza virus activity of diphenhydramine hydrochloride

[0033] 1. Experimental materials and methods

[0034] 1.1 Cells, viruses and drugs

[0035] MDCK cells were purchased from ATCC, virus A / PR8 / 34H1N1 was amplified from chicken embryo culture, and diphenhydramine hydrochloride was purchased from Sigma;

[0036] 1.2 Experimental Instruments

[0037] Multifunctional detector PerkinElmer, inverted microscope Costar

[0038] 2. experimental method:

[0039] 2.1 Cell culture:

[0040] 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.

[0041] 2.2 Virus culture:

[0042] Take 9-11 day-old SPF chicken embryos, check them with an egg tester before inoculating the virus, and mark them at a posit...

Embodiment 2

[0057] Embodiment 2: the inhibitory effect of diphenhydramine hydrochloride on different stages of virus 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 ...

Embodiment 3

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

[0063] In this experiment, the antiviral activity of diphenhydramine hydrochloride on influenza A virus subtype H1N1 strain, influenza A virus H3N2 strain and influenza B virus strain was detected.

[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 cell culture incubator at 37°C 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, add 100TCID50 / well virus solution to infect the cells, and add drugs of various concentration gradients at the same time (starting at 200μM, serially dilute 6 gradients by 2 times, each gradient 2 A total of 200 μl of maintenance medium (DMEM+0.3%BSA+2.5ug / L...

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Abstract

The invention discloses an application of diphenhydramine hydrochloride in preparation of medicines for treating or preventing influenza viruses. Firstly, diphenhydramine hydrochloride with completely nontoxic concentration is selected for an antiviral test, and the result shows that the small molecular compound has remarkable antiviral activity and is in dose-dependent association; then, the action period of diphenhydramine hydrochloride which resists influenza viruses is analyzed, and the result shows that diphenhydramine hydrochloride mainly inhibits events in early stage of virus infection such as virus absorption and entrance; and finally, the antiviral activity of diphenhydramine hydrochloride to different types and sub-types of influenza viruses is detected, and the result shows that diphenhydramine hydrochloride can inhibit replication of all detection viral strains and has the dose-dependent effect, thereby representing that diphenhydramine hydrochloride is of certain broad spectrum to anti-influenza virus activity. The diphenhydramine hydrochloride disclosed by the invention can be developed as a novel anti-influenza virus medicine and provides a novel path and means for treating influenza.

Description

technical field [0001] The invention belongs to the technical field of medicine, and more specifically relates to the application of diphenhydramine hydrochloride 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 (P...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/135A61P31/16
Inventor 陈绪林廖庆姣
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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