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Broad-spectrum antiviral drug or composition

A technology of antiviral drugs and compositions, applied in the field of broad-spectrum antiviral drugs or compositions, can solve the problems of high viral infectivity, no specific treatment drugs, huge harm to aquaculture and human health, etc. Broad-spectrum and high-efficiency antiviral activity, the effect of ensuring sustainable development and human health

Active Publication Date: 2019-11-05
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Viruses such as PEDV, PRV, and HSV-1 are highly contagious and pose great hazards to the breeding industry and human health. At present, most of them have no specific therapeutic drugs

Method used

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  • Broad-spectrum antiviral drug or composition
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  • Broad-spectrum antiviral drug or composition

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0051] 1. Preparation of cells

[0052] The DMEM medium containing 10% fetal bovine serum and 100U penicillin and streptomycin was used for Vero cell passage, and the DMEM medium containing 2% fetal bovine serum and 100U penicillin streptomycin was used as the maintenance solution.

[0053] 2. Virus proliferation

[0054] Proliferation of HSV-1, NDV, VSV, and PRV cytotoxicity: Inoculate the virus stock solution according to the amount of 1 / 10 of the cell flask culture medium, and wait for the cytopathic changes (such as NDV to form large syncytia) to reach 75% (usually 48 hours) to use repeatedly Harvest the virus by freezing and thawing, that is, store the cell bottle at -80°C and store it at room temperature until it partially thaws. The virus was released from the cells and frozen at -80°C.

[0055] Proliferation of NDV chicken embryo virus: 9-10 day-old chicken embryos were inoculated with NDV virus stock solution through the allantoic cavity, and the allantoic fluid was...

Embodiment 1

[0107] Example 1 Effect of RAF265 on PRV virus virulence

[0108] Infect Vero cells with PRV Fa strain at 0.1 MOI, add different doses of RAF265 (0, 10, 25, 50 μM) (prepared in DMSO, the same below) at the same time during virus infection, incubate for 1 hour, discard the supernatant, wash with PBS 2 times, add cell maintenance solution, culture in a cell incubator for 24 hours, and observe cell lesions. The results showed that RAF265 could inhibit the cytopathic effect caused by PRV in a dose-dependent manner ( Figure 1A ).

[0109] Infect Vero cells with 0.1 MOI of PRV Fa strain, add different doses of RAF265 (0, 5, 10, 25, 50 μM) at the same time of virus infection, after incubation for 1 h, discard the supernatant, wash 2 times with PBS, add cells to maintain The cells were cultured in a cell incubator for 24 hours, the cells were collected, and the expression of the viral protein gC protein was detected by immunoblotting, or the virus virulence was detected after repeat...

Embodiment 2

[0114] Example 2 Effect of RAF265 on virulence of HSV-1 virus

[0115] Vero cells were infected with 0.1MOI GFP-HSV-1 virus (Xin-Jing L et al., 2011). At the same time, three RAF inhibitors were added, namely RAF265, CEP32496 and BGB283 (CAS No. 927880-90-8 , 1188910-76-0 and 1446090-79-4 were all dissolved with DMSO), after 1 hour of action, the supernatant was discarded, and washed twice with PBS, cultivated in a cell culture box for 24 hours, and observed the virus with a fluorescence microscope Olympus IX73 Replicate the situation, and collect the cells for western blotting. The results showed that among the three inhibitors, RAF265 had the best effect on inhibiting the virus, and at a dose of 25 μM, almost no virus particles could be observed in the fluorescence microscope ( Figure 2A ). Similarly, in Western blot experiments, compared with the other two inhibitors, 25 μMRAF265 can inhibit the expression of HSV-1ICP8 protein to a level that is almost undetectable ( F...

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Abstract

The invention provides a broad-spectrum antiviral drug or composition. The drug or composition is prepared from the active components of a compound RAF265 or a medically acceptable salt thereof. According to the broad-spectrum antiviral drug or composition, the RAF265 is first found to have the broad-spectrum high-efficiency antiviral activity, 7 types of viruses (PRV, HSV-1, PEDV, FMDV, NDV, VSVand IBDV) of 6 families can be efficiently inhibited from infecting host cells and mice, the amount of the viruses in the mice is significantly decreased, and the symptoms of infection are relieved; and the RAF265 has the low cytotoxicity (CC50 / IC50 is greater than 100), and the toxic and side effects on chick embryos and the mice are avoided. According to the broad-spectrum antiviral drug or composition, a provided kinase inhibitor capable of efficiently resisting multiple viral infections has broad application prospects in the fields of aquaculture and biomedicine.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a broad-spectrum antiviral drug or composition. Background technique [0002] In recent years, the research on broad-spectrum antiviral agents has made great progress, breaking through the bottleneck and limitation of a single host of antiviral agents and the relatively weak activity of broad-spectrum agents, and some achievements are already facing application. The research on antiviral preparations is based on two aspects of design, one is from the level of virus infection, and the other is from the level of host cell defense. Antiviral preparations are studied from the perspective of host cell defense, that is, by changing or eliminating proteins or molecules that are required for virus replication but not important for the growth of the cell itself, to affect the life cycle of the virus, so as to achieve antiviral effects. Antiviral agents targeting host cells can truly achieve br...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4439A61P31/22A61P31/14
CPCA61K31/4439A61P31/22A61P31/14
Inventor 王晓佳薛彦李翠翠杨德吉路金金
Owner CHINA AGRI UNIV
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