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Application of indole-3-acetonitrile in preparation of medicine for treating or preventing influenza virus infection

A technology for influenza virus infection and influenza prevention, applied in the field of biomedicine, can solve problems such as unreported anti-influenza virus function, and achieve the effects of easy large-scale production, low cytotoxicity, and economical speed.

Active Publication Date: 2021-02-12
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on indole-3-acetonitrile is limited to plant growth hormone and the inhibitory effect on some bacterial biofilms, and there is no report on the anti-influenza virus function

Method used

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  • Application of indole-3-acetonitrile in preparation of medicine for treating or preventing influenza virus infection
  • Application of indole-3-acetonitrile in preparation of medicine for treating or preventing influenza virus infection
  • Application of indole-3-acetonitrile in preparation of medicine for treating or preventing influenza virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Effect of indole-3-acetonitrile on cell viability:

[0030] The effect of indole-3-acetonitrile on cell viability was determined by CCK-8 kit, and the optimal experimental concentration was screened out. details as follows:

[0031] The A549 cells in the 96-well plate grow to about 80%, incubate different concentrations (12.54 μM, 125.4 μM, 300 μM, 350 μM, 376.2 μM) of indole-3-acetonitrile, and cultivate them in a CO2 incubator at 37 ° C for 24 hours, and then protect them from light. Then add CCK-8 (10 μL / well). After culturing in a 37°C CO2 incubator for 2 hours, use a microplate reader to measure the OD 450nm absorbance at .

[0032] The results showed that the cell activity incubated with 376.2 μM indole-3-acetonitrile was 84.9%, and the cell activity incubated with 350 μM indole-3-acetonitrile was 98.8% (Table 1). Since there was no statistical difference between 98.8% and 100%, , 350 μM indole-3-acetonitrile was used for subsequent experimental studies.

[0...

Embodiment 2

[0037] Preliminary screening of the inhibitory effect of indole-3-acetonitrile on the proliferation of influenza A virus in vitro:

[0038] 1. Fluorescence experiment.

[0039] h 5 N 6 - After GFP virus infects cells, GFP fluorescence will increase with the proliferation of influenza virus. By observing the intensity of GFP fluorescence, it can indirectly reflect the proliferation of influenza virus. When the compound inhibits the proliferation of influenza virus, GFP fluorescence will be weakened. details as follows:

[0040] A549 in a 12-well plate was cultured in a CO2 incubator at 37°C until a cell monolayer was formed, and infected with H at 0.01MOI 5 N 6 -GFP, after 1h, they were replaced with different compounds (indole-3-acetonitrile, indole-3-carbaldehyde (Indole-3-carboxaldehyde), gramine (Gramine), indole-3-carboxylic acid (3-Indoleformic acid) acid)) cell maintenance solution (2% serum F-12), after 24 hours, the fluorescence was observed under an inverted micr...

Embodiment 3

[0047] Effects of indole-3-acetonitrile on viral mRNA and host mRNA:

[0048] In order to study the effect of indole-3-acetonitrile on mRNA, a fluorescence quantitative test was carried out, as follows:

[0049] First, extract RNA and convert it into cDNA, use Trizol reagent to fully lyse the cells, then transfer to RNase-free EP tubes, fully invert, and ice-bath for 10 minutes. Add 200 μL of chloroform to each tube, vortex and centrifuge. Take the supernatant to a new RNase-free EP tube, add an equal volume (500 μL) of isopropanol, mix gently, let stand and centrifuge, and discard the supernatant. Add 900 μL of ethanol, vortex and centrifuge, discard the supernatant. Blow dry while working on the clean bench. Add the DNA digestion system, bathe in a water bath at 37°C for 1 hour, then add 5 μL of DNAStop Solution to each tube, flick it briefly, and then bathe in water at 65°C for 15 minutes. Take a small amount to measure the RNA concentration. After the preparation of t...

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Abstract

The invention belongs to the field of biological medicines, and particularly discloses an application of indole-3-acetonitrile in preparation of a medicine for treating or preventing influenza virus infection. The applicant finds that indole-3-acetonitrile can improve weight loss caused by infection of influenza viruses, remarkably inhibit proliferation of the influenza viruses in a mouse body andimprove lung tissue lesion caused by infection of the influenza viruses, and has an obvious anti-influenza virus infection effect. Meanwhile, indole-3-acetonitrile can inhibit proliferation of various influenza A viruses including human influenza virus PR8, swine influenza virus G4 and avian influenza virus H5N6, has broad spectrum, is simple in synthesis process, economical and rapid, is easy toproduce on a large scale, and can be developed into anti-influenza virus drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of indole-3-acetonitrile in the preparation of medicines for treating or preventing influenza virus infection. Background technique [0002] Influenza virus (influenza virus) referred to as influenza virus, including human influenza virus and animal influenza virus, can cause human, poultry, pig, horse, bat and other animal infection and disease, human influenza virus is divided into A (A), B (B), C (C), D (D) four types. Among them, influenza A virus spreads rapidly, easily spreads from person to person, and even spreads across species. Because the antigenicity of influenza virus is easy to mutate, it has repeatedly caused worldwide pandemics. The influenza pandemic has brought heavy disasters to human health and the global economy. [0003] Currently clinically used western medicine anti-influenza drugs are mainly divided into neuraminidase (NA) inhibit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/404A61P31/16A61P11/00
CPCA61K31/404A61P31/16A61P11/00
Inventor 金梅林赵雪锦黄坤赵亚张强盛锋杜雪竹龚文孝邹忠李再云杨丽孙小美
Owner HUAZHONG AGRI UNIV
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