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Application of GW3965 in preparation of drugs treating or preventing hepatitis C

A hepatitis C virus and drug technology, applied in the field of medicine, can solve the problem of no application of GW3965 in anti-hepatitis C virus, and achieve the effect of improving the cure rate

Inactive Publication Date: 2013-04-24
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently no applications of GW3965 against HCV

Method used

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  • Application of GW3965 in preparation of drugs treating or preventing hepatitis C
  • Application of GW3965 in preparation of drugs treating or preventing hepatitis C
  • Application of GW3965 in preparation of drugs treating or preventing hepatitis C

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Evaluation of GW3965 anti-hepatitis C virus activity

[0036] 1. Experimental materials

[0037] 1.1 Cells, plasmids, viruses and drugs

[0038] Huh7.5.1 cells were donated by Dr.F.V.Chisari; plasmid pJFH1 containing the complete genome sequence of HCV type 2a JFH1 strain was donated by Prof. Dr. Takaji Wakita; plasmid pEGFP-N1 was purchased from Clontech; plasmid pGL4.70[hRLuc] was purchased from promega company; the virus JFH1-Luc-5AGFP with double reporter genes was prepared by our laboratory; GW3965 was purchased from sigma company.

[0039] 1.2 Reagents

[0040] DMEM medium was purchased from GIBCO Company; Renilla luciferase detection kit was purchased from Promega Company; anti-HCV NS3 mouse monoclonal antibody was purchased from Henan Bioengineering Technology Research Center; anti-GAPDH mouse monoclonal antibody was purchased from Zhongshan Jinqiao, Beijing Company; HRP-labeled goat anti-mouse secondary antibody was purchased from Thermo Company; ...

Embodiment 2

[0059] Embodiment 2: The research of GW3965 enters the influence of hepatitis C pseudovirus (HCVpp)

[0060] 1. Experimental materials

[0061] 1.1 Cells, viruses and drugs

[0062] Huh7.5.1 cells; GW3965; plasmid PNL4.3-R-E-Luc and plasmid pVpack-VSV-G containing the VSV viral envelope protein were a gift from the Division of AIDS Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health. The plasmids containing HCV1a and 1b envelope proteins were donated by Mr. Qi Zhongtian.

[0063] 1.2 Reagents

[0064] DMEM medium was purchased from GIBCO; transfection reagent Lipofectamine2000 was purchased from Invitrogen; Firefly luciferase detection kit was purchased from Promega.

[0065] 1.3 Experimental Instruments

[0066] The 20 / 20 detector is a product of Promega.

[0067] 2. Experimental methods and results

[0068] 2.1 Compared with the live virus, the HCV pseudovirus has similar cell infection characteristics, which can be used to ...

Embodiment 3

[0070] Example 3: Combination study of GW3965 and other drugs with anti-HCV activity.

[0071] 1. Experimental materials

[0072] 1.1 Cells, viruses and drugs

[0073] Huh7.5.1; virus JFH1-Luc-5AGFP; GW3965; CsA (purchased from Sigma Company) and MK-7009 (purchased from Zannan Company).

[0074] 1.2 Reagents

[0075] DMEM medium was purchased from GIBCO; Renilla luciferase detection kit was purchased from Promega.

[0076] 1.3 Experimental Instruments

[0077] The 20 / 20 detector is a product of Promega.

[0078] 2. Experimental methods and results

[0079] 2.1 Divide Huh7.5.1 cells into 8×10 cells 3 Each cell / well was inoculated in a 96-well cell culture plate, cultured in a 37°C cell culture incubator for 14-18 hours, and then used after the cells grew into a monolayer. Cyclosporine A (CsA) or MK-70092-fold gradient dilution was added to the orifice plate as a control group for separate medication, and each group had 3 repetitions; in addition, 500nM GW3965 was added...

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Abstract

The invention discloses application of GW3965 (3-[3-[[[2-chloro-3-(trifluoromethyl)phenyl]methyl](2, 2)-biphenylethyl)amino]propoxy)phenylacetate hydrochloride) in preparation of drugs treating or preventing hepatitis C virus. A drug without any toxic concentration is chosen for an antiviral experiment, the influence of GW3965 on virus gene duplication is detected, and results show that the small molecule compound has significant antiviral activity and is dose dependent. Then, the GW3965 is employed to treat the hepatitis C pseudovirus system of different gene subtypes, and results show that the compound inhibits the virus entrance link at the early stage of a virus lifecycle. The anti-virus capability of drug combination of GW3965 and other anti-HCV drugs is detected, and results show that the compound has the potential of drug combination with other drugs. GW3965 can inhibit 1a, 1b, and 2a genotype virus entrance, and has broad-spectrum antiviral activity and a novel target, while there is no anti-hepatitis C virus drug with entrance as the target clinically. And drug combination can achieve a good effect.

Description

technical field [0001] The present invention belongs to the technical field of medicine, and more specifically relates to a small molecule compound GW3965 (3-[3-[[[2-chloro-3-(trifluoromethyl)phenyl]methyl](2,2-bi Phenylethyl) amino] propoxy] phenylacetic acid hydrochloride) in the preparation of medicines for treating or preventing hepatitis C virus (HCV). Background technique [0002] Hepatitis C Virus (HCV), discovered in 1989, is the main pathogen causing non-A, non-B hepatitis after blood transfusion. About 170 million people worldwide are currently infected with HCV. HCV infection is distributed worldwide, the infection rate is about 3%, and 3 to 4 million new cases are added every year. According to the serological epidemiological survey data in my country, the positive rate of HCV in the general population is 3.2%. The main routes of transmission of HCV include blood transfusion or blood products, intravenous drug use, sexual contact, and mother-to-child transmiss...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/195A61P1/16A61P31/14
Inventor 陈绪林曾晶廖庆姣吴阳
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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