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Application of triparanol to resisting Ebola virus infection

A technology of Ebola virus and tripalarol, applied in the field of medicine

Inactive Publication Date: 2017-02-01
SUZHOU INST OF SYST MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

EBOV vaccines are mainly divided into three categories based on antigen delivery methods, including vaccines based on non-replicating viral vectors, vaccines based on replicating viral vectors, and vaccines based on viral protein antigens, among which, GlaxoSmithKline and the National Institute of Allergy and Infectious Diseases The ChAd3-ZEBOV jointly developed by the Institute and the VSV-EBOV hybrid vaccine developed by the Public Health Agency of Canada have the most potential and have entered the clinical research stage, but more studies are still needed to prove their effectiveness

Method used

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  • Application of triparanol to resisting Ebola virus infection
  • Application of triparanol to resisting Ebola virus infection
  • Application of triparanol to resisting Ebola virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: EBOV GP-HIV system screening principle

[0020] The entry of Ebola virus into the host cell is the first step of virus infection, and inhibiting the entry of the virus can effectively block the virus infection. EBOV envelope surface glycoprotein (Glycoprotein.GP) is a key protein that mediates Ebola virus adsorption and entry into target cells.

[0021] We synthesized the envelope GP gene of Zaire Ebola virus (EBV-Zaire GP, Gene Accesion No.L11365). By co-transfecting EBV-GP and pNL4-3.Luc.R - E. - Co-transfect cells to obtain EBV recombinant virus EBV-GP / HIV with EBV-GP as the HIV core wrapped in the shell. The virus particle has the following characteristics: 1) The selectivity of the virus to the host cell depends on the characteristics of EBOV-GP; 2) Due to the deletion of the env, nef and vpr genes on the HIV vector, the virus can only enter the host cell at one time and Cannot replicate, so the virus is safe; 3) The HIV vector has a luciferase report...

Embodiment 2

[0022] Embodiment 2: Utilize VSV GP / HIV and EBOV-GP / HIV pseudovirus systems to evaluate the antiviral effect of tripalarol respectively.

[0023] In the present invention, EBV-Zaire (Gene Accession No. L11365) is used to evaluate the pharmacological activity of trippalatol against Ebola virus infection.

[0024] Recombinant virus preparation: co-transfect 2ug pcDNA3.1 / EBV-GP plasmid and 20ug pNL4-3.Luc.

[0025] R - E. - The plasmid was transferred to 293T cells, and the supernatant was collected after 48 hours of transfection. The supernatant was filtered through a 0.45uM filter. The supernatant contained EBV-GP / HIV virus particles, and the recombinant virus could be used for infection. Prepare VSV-G / HIV recombinant virus according to the same method.

[0026] Infection and detection: the day before infection, 10 per well 4 Hela cells were seeded on a 96-well plate at a density of 1 cell. Dissolve the positive control compound or the compound to be screened in ethanol, a...

Embodiment 3

[0027] Embodiment 3: Cytotoxicity detection test

[0028] The MTS method was used to measure the cytotoxicity of all the involved drugs to Hela cells in our experimental system, and the results showed that tripalarol had no cytotoxicity at a final concentration of 5uM.

[0029] We use the Ebola virus reporter system (EBOV GP-HIV pseudovirus system) that can be researched in the P-2 laboratory to evaluate a variety of small molecular compounds. It has obvious inhibitory effect when invading infection. Tripalarol is a cholesterol-lowering drug listed in the 1960s, which inhibits 24-dehydrocholesterol synthesis reductase (DHCR24: is located in the endoplasmic reticulum flavin adenine dinuclear Glycine-dependent oxidoreductase, which catalyzes the reduction of sterol δ-24 double bonds to single bonds, and synthesizes cholesterol) reduces the synthesis of cholesterol in cells, thereby feedback stimulating low-density lipoprotein (low density lipoprotein) on the surface of cell memb...

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Abstract

The invention provides an application of triparanol to resisting Ebola virus infection, and particularly relates to an application of triparanol to blocking Ebola invasion infection. A theoretical basis is provided for developing an antiviral combined drug taking the triparanol as a main component, and technical support is provided for prevention and control of viral emerging infectious diseases.

Description

technical field [0001] The invention discloses the application of tripalamol in blocking Ebola virus invasion and infection, and belongs to the technical field of medicine. Background technique [0002] Ebola hemorrhagic fever is an acute hemorrhagic infectious disease caused by Ebola virus (EBOV). It is mainly infected through contact with the blood, body fluids, secretions and excreta of patients or infected animals. The main clinical manifestations are prominent fever, bleeding and multiple organ damage. The Ebola virus epidemic that broke out in 2014 swept across West Africa, and the extremely high fatality rate caused panic all over the world. [0003] Ebola virus (EBOV) belongs to the Filoviridae family and is filamentous. It is a single-stranded negative-sense RNA virus with 18959 bases and an envelope. Pure virions consist of a helical ribose-nucleocapsid complex, containing negative-strand linear RNA molecules and four virion structural proteins. The Ebola viral ...

Claims

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Application Information

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IPC IPC(8): A61K31/138A61P31/14
CPCA61K31/138
Inventor 秦晓峰左向阳吴飞
Owner SUZHOU INST OF SYST MEDICINE
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