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Application of cellular target liver X receptor in preparation of drugs treating hepatitis C virus

A hepatitis C virus and target technology, applied in the field of medicine, to 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 is currently no application of LXR as a target for screening anti-HCV drugs

Method used

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  • Application of cellular target liver X receptor in preparation of drugs treating hepatitis C virus
  • Application of cellular target liver X receptor in preparation of drugs treating hepatitis C virus
  • Application of cellular target liver X receptor in preparation of drugs treating hepatitis C virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Evaluation of the anti-hepatitis C virus activity of the LXR agonists GW3965 and T0901317.

[0042] 1. Experimental materials

[0043] 1.1 Cells, plasmids, viruses and drugs

[0044] 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 and T0901317 were purchased from sigma company.

[0045] 1.2 Reagents

[0046] 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 Beijing Zhongshan Golden Bridge Company; HRP-labeled goat anti-mouse secondary ...

Embodiment 2

[0065] Example 2: Study on the Effect of LXR Agonist GW3965 and T0901317GW3965 on the Entry of Hepatitis C Pseudovirus (HCVpp)

[0066] 1. Experimental materials

[0067] 1.1 Cells, viruses and drugs

[0068] Huh7.5.1 cells; GW3965 and T0901317; plasmid PNL4.3-R-E-Luc and plasmid pVpack-VSV-G containing the VSV viral envelope protein were obtained from the AIDS Research Division of the 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.

[0069] 1.2 Reagents

[0070]DMEM medium was purchased from GIBCO; transfection reagent Lipofectamine2000 was purchased from Invitrogen; Firefly luciferase detection kit was purchased from Promega.

[0071] 1.3 Experimental Instruments

[0072] The 20 / 20 detector is a product of Promega.

[0073] 2. Experimental methods and results

[0074] 2.1 Compared with the live virus, the HCV pseudovirus has similar cell infe...

Embodiment 3

[0076] Example 3: Combination study of LXR agonists GW3965 and T0901317 with other drugs with anti-HCV activity.

[0077] 1. Experimental materials

[0078] 1.1 Cells, viruses and drugs

[0079] Huh7.5.1; virus JFH1-Luc-5AGFP; GW3965 and T0901317; CsA (purchased from Sigma Company) and MK-7009 (purchased from Zannan Company).

[0080] 1.2 Reagents

[0081] DMEM medium was purchased from GIBCO; Renilla luciferase detection kit was purchased from Promega.

[0082] 1.3 Experimental Instruments

[0083] The 20 / 20 detector is a product of Promega.

[0084] 2. Experimental methods and results

[0085] 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 well plate as a control group for separate medication, and each group had 3 repetitions...

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Abstract

The invention discloses application of a cellular target liver X receptor in preparation of drugs treating hepatitis C virus (HCV), specifically a cellular target that can be used for screening anti-HCV drugs. The cellular target is a liver X receptor (LXR). The agonist GW3965 and T0901317 of the liver X receptor have significant anti-hepatitis C activity in invitro anti-hepatitis C virus experiments. The cellular target liver X receptor can be used for screening drugs treating the hepatitis C virus. Therefore, the cellular target LXR involved in the invention can be applied to drug development as a new anti-hepatitis C virus target, thus providing a new approach and means for treatment and cure of hepatitis C.

Description

technical field [0001] The invention belongs to the technical field of medicine, and more specifically relates to the application of a cell target liver X receptor in the preparation of medicines for treating or preventing hepatitis C virus. 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 transmission. HCV infection can easily lead to chronicity, only about 20% of HCV-infected patients can heal spontaneously, and ...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N21/64
Inventor 陈绪林曾晶廖庆姣吴阳
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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