Applications of cardiac glycoside compound in resisting Ebola virus infection

A technology of Ebola virus and cardiac glycosides, applied in antiviral agents, medical preparations containing active ingredients, organic active ingredients, etc., to achieve the effect of blocking invasion and inhibiting replication

Inactive Publication Date: 2017-04-19
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 Institut...

Method used

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  • Applications of cardiac glycoside compound in resisting Ebola virus infection
  • Applications of cardiac glycoside compound in resisting Ebola virus infection
  • Applications of cardiac glycoside compound in resisting Ebola virus infection

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0034] Embodiment 1: Screening model principle

[0035] 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. 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 env, nef and vpr genes on the HIV vector, the virus can only enter the host cell at one time and Can not replicate, so the virus is safe; 3) The HIV vector has a luciferase reporter gene, so the infect...

Embodiment 2

[0038] Embodiment 2: Utilize the Ebola pseudovirus reporter system (EBVG / HIV-Luc) to evaluate the effect of digoxin, digitoxin, lanatoside C, and oleuroside G in Ebola virus invasion infection

[0039] Recombinant virus preparation: co-transfect 2ug pcDNA3.1 / EBV-GP plasmid and 20ug pNL4-3.Luc.R - E. - Put the plasmid into 293T cells, collect the supernatant after 48 hours of transfection, and filter the supernatant through a 0.45uM filter membrane. The supernatant contains EBV-GP / HIV virus particles, and the recombinant virus can be used for infection.

[0040] Infection and detection: the day before infection, 10 per well 4Hela 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 methanol, add it to the cell culture medium 15 minutes before infection, use methanol as a blank control, add 0.1ul of EBV-GP / HIV-luc virus solution (10 9 rluc / ml) infected cells, 48 ​​hours after infection, discard t...

Embodiment 3

[0041] Example 3: Using the Ebola Minigenome System (EBOV-Minigenome System) to evaluate the effects of digoxin, digitoxin, ouanthaside C, and ouanthaside G in the transcription and replication of Ebola virus The implementation is as follows:

[0042] 10 per well the day before transfection 4 293T cells were plated in a 96-well plate, and the next day, a small replicon system was used at 200ng per well (NP: 12.5ng; VP35: 12.5ng; VP30: 7.5ng; L: 100ng; MINI-Rluc: 25ng; T7: 50ng) Carry out transfection (wherein each well is transfected with 2ng internal reference plasmid), after 24h, different concentration gradients (500nM; 250nM; 125nM; 62.5nM; 31.25nM, 0nM) were added digoxin, digoxigenin, trichophyllin Glycoside C; incubation with elatoside G (methanol as the control group); after 8 hours, remove the supernatant and replace with a new medium, and after another 24 hours, use the dual luciferase reporter gene detection kit to detect, the results show that Digoxin, Digitoxin,...

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Abstract

The invention discloses applications of cardiac glycoside medicines in resisting Ebola virus infection. The infection stages comprise invasion, transcription and replication; the cardiac glycoside medicines comprise but are not limited to digoxin, digitoxin, lanatoside C and ouabain G. The four representative cardiac medicines can effectively prevent the Ebola virus from infecting host cells, thus a theoretical basis is provided for developing the antiviral compound medicines taking the cardiac glycoside as the main ingredient, and meanwhile, a scientific and technical support is provided for preventing and controlling the viral emerging infectious diseases.

Description

technical field [0001] The present invention belongs to the field of new application of compounds, and specifically relates to the application of cardiac glycosides in anti-Bora virus infection, wherein cardiac glycosides include but not limited to digoxin, digitoxin, lanatoside C and toxin Electraside G. 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 heli...

Claims

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Application Information

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IPC IPC(8): A61K31/7048A61P31/14
CPCA61K31/7048
Inventor 秦晓峰左向阳吴飞
Owner SUZHOU INST OF SYST MEDICINE
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