Application of dendrobine in preparation of anti-influenza virus medicines

An anti-influenza virus and influenza virus technology, applied in the field of medicine, can solve problems such as unreported application of anti-influenza virus, and achieve the effects of reducing immune cross-protection effect, solving drug resistance, and broad-spectrum antiviral activity

Active Publication Date: 2016-11-23
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dendrobine belongs to the sesquiterpene alkaloids, with a five-membered and six-membered ring structure containing a nitrogen atom as the mother core. Its structure is as follows. It can lower heart rate, blood pressure, and slow breathing, but its application in anti-influenza virus has not been reported

Method used

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  • Application of dendrobine in preparation of anti-influenza virus medicines
  • Application of dendrobine in preparation of anti-influenza virus medicines
  • Application of dendrobine in preparation of anti-influenza virus medicines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The cytotoxicity detection of embodiment 1 dendrobine

[0037] The cytotoxicity of dendrobine was detected by MTT method. The specific method is as follows:

[0038] MDCK or A549 cells by 1×10 4 / well seeded in 96-well plate at 37°C, 5% CO 2 Cultured in a constant temperature cell incubator until monolayer, DMEM or 1640 gradiently diluted dendrobine was added to a 96-well plate, 200 μl per well, and continued to culture for 48 hours. Discard the culture supernatant, add 100 μL of 1640 or DMEM medium containing 0.5 mg / ml MTT to each well, and incubate at 37°C for 4 hours. The absorbance at 570 nm was detected with a multifunctional microplate reader (Genios Pro, Tecan, US). The survival rate of the cells was used as the indicator of the toxicity of dendrobine to MDCK or A549 cells.

[0039] Cell viability (%)=E / N×100

[0040] E is the absorbance of the drug group, and N is the absorbance of the cell control group.

[0041] Test results: Dendrobine has low cytotoxi...

Embodiment 2

[0043] Example 2 Detection of Dendrobine Anti-Influenza A Virus Activity in Vitro

[0044] In the in vitro antiviral experiment of the present invention, various subtypes of influenza A viruses are involved, including H1N1 and H3N2, and the specific methods are as follows:

[0045] MDCK cells by 2 x 10 4 / well seeded in 96-well plate at 37°C, 5% CO 2 cultured to a monolayer in a constant temperature cell incubator. Use 100TCID 50 Infect the cells with influenza A virus, 100 μl per well, incubate at 37°C for 1 h, discard the virus solution, add dendrobine diluted in DMEM (containing 1 μg / ml TPCK), 200 μl per well, and continue to incubate for 48 h. The antiviral activity of dendrobine is determined by the protective effect of dendrobine on cells, including observing the cell virus phenomenon (CPE) caused by dendrobine inhibiting virus and detecting the survival rate of cells, and further calculating the half effective concentration IC 50 . Ribavirin (ribavirin) was used as...

Embodiment 3

[0050] Example 3 Dendrobine inhibits the replication of influenza A virus

[0051] In order to evaluate the inhibitory effect of dendrobine on influenza virus replication, the present invention adopts two methods of Q-PCR and Western blotting to detect the influence of dendrobine on virus replication from the expression levels of genes and proteins respectively. The specific method is as follows:

[0052] MDCK cells by 2 x 10 4 / well seeded in a 6-well plate at 37 °C, 5% CO 2 cultured to a monolayer in a constant temperature cell incubator. Use 100TCID 50 Infect the cells with influenza A virus, 1ml per well, incubate at 37°C for 1h, discard the virus liquid, add dendrobine diluted in DMEM (containing 1μg / ml TPCK), 2ml per well, and continue to incubate for 24h. After that, cells were fully lysed with 1ml TRIzol and total RNA was extracted, and samples were used for Q-PCR; while cells were lysed with 80 μl RIPA lysate containing protease and phosphatase inhibitors on ice a...

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Abstract

The invention discloses application of dendrobine in preparation of anti-influenza virus medicines. The influenza virus is influenza a virus, preferably H1N1, H3N2, H5N1, H7N1, H7N2, H7N3, H7N7, H7N9, H9N2 or H10N8. The dendrobine can be used for preparing medicines capable of inhibiting replication of influenza a virus in host cells, and is capable of inhibiting denucleation of a virus ribonucleoprotein complex vRNP or activity of the ribonucleoprotein complex vRNP, wherein loci at which the dendrobine is combined with influenza virus NP proteins are Arg267 at locus 267, Val270 at locus 270, Phe338 at locus 338 and Glu339 at locus 339.

Description

technical field [0001] The invention relates to the application of dendrobine in anti-influenza virus drugs, and belongs to the technical field of medicine. Background technique [0002] Influenza viruses belong to the single-stranded negative-sense RNA virus of the Orthomyxoviridae family and are divided into three subtypes: A (A), B (B) and C (C). Influenza A virus is prone to antigenic variation, adaptation and recombination, which will form new highly lethal strains and cause a worldwide pandemic. In addition to the great damage caused by several influenza outbreaks in history to human beings and society, new influenza viruses that have emerged in recent years, such as H5N1, H7N9, H9N2, H5N6, etc., have also seriously threatened human health. Therefore, effective methods for preventing and treating influenza are urgently needed. Early detection and early treatment can obtain better curative effect. For this, anti-influenza virus drugs have more advantages than vaccine...

Claims

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Application Information

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IPC IPC(8): A61K31/407A61P31/16
CPCA61K31/407
Inventor 杨洁刘叔文李日婵
Owner SOUTHERN MEDICAL UNIVERSITY
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