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Use of compound MEAN in preparation of medicine for inhibiting HCV replication

A technology of compounds and drugs, applied in the field of medicine, can solve problems such as unreported antiviral activity

Inactive Publication Date: 2015-01-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Compound MEAN (6-methoxyethylamino-numonafide) is a newly synthesized class of numonafide compounds. The synthetic route of this compound has been reported in the literature (John T.Norton, Anti-Cancer Drugs 2008, 19 (11)), but its antiviral activity has not been reported. According to the report, the present invention provides the new application of compound MEAN (6-methoxyethylamino-numonafide) anti-HCV activity

Method used

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  • Use of compound MEAN in preparation of medicine for inhibiting HCV replication
  • Use of compound MEAN in preparation of medicine for inhibiting HCV replication
  • Use of compound MEAN in preparation of medicine for inhibiting HCV replication

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 drug cytotoxicity analysis

[0028] cell culture

[0029] Huh-7.5.1 cells were cultured in DMEM medium containing 10% fetal bovine serum in an incubator at 37°C and 5% CO2 saturated humidity. 100U / ml of penicillin and 100ug / ml of streptomycin were added to the culture medium.

[0030] Preparation of HCV RNA Transcripts

[0031] Take 15 μg of pFL-JC1 plasmid, carry out enzyme digestion reaction with restriction endonuclease XbaI, incubate at 37°C for 2 hours, and then check whether it is completely linearized by agarose gel electrophoresis. The digested products were digested with mung bean nuclease and proteinase K, and then the DNA was extracted and purified by the phenol-chloroform method, and then detected and quantified by agarose gel electrophoresis. Then 2 μg of DNA template was taken, and T7 in vitro transcription kit was used for in vitro transcription according to the instructions. The obtained RNA transcripts were detected by agarose gel elect...

Embodiment 2

[0037] Embodiment 2 MEAN anti-HCV efficacy analysis

[0038] Analysis of anti-HCV effect in vitro

[0039] Huh7.5.1 cells were inoculated at 5000 per well in a 96-well plate and incubated overnight. The wild-type virus JC1-luc was used to infect the cells at MOI0.01 for 24 hours, and then given 0.625 μM, 1.25 μM, 2.5 μM, 5 μM MEAN, and 48 hours later, micro Luciferase activity was quantified with a plate luminometer (Turner Biosystems). The inhibitory activity of MEAN was verified by monitoring the expression levels of HCV RNA and protein in Huh7.5.1 cells infected with the virus.

[0040] Fluorescent quantitative PCR

[0041] Fluorescent quantitative PCR was used to detect the relative amount of HCV RNA in the cells. Cells were collected after trypsinization, and Trizol was added to each tube to extract total RNA according to the operating instructions, and the concentration was measured by an ultraviolet spectrophotometer. Take 4 μl of RNA and remove the genomic DNA acco...

Embodiment 3

[0048] Embodiment 3 MEAN anti-E2 mutant strain JC1 / N415D or JC1 / G451R effect

[0049] Anti-E2 mutant strain JC1 / N415D or JC1 / G451R effect analysis in vitro

[0050] Huh7.5.1 cells were inoculated in 96-well plates at 5000 per well and incubated overnight. The cells were infected with the mutant virus JC1 / N415D or JC1 / G451R at MOI0.01 for 24 hours, and MEAN was given at a concentration gradient. After 48 hours, a microplate luminometer was used to (Turner Biosystems) to quantify luciferase activity.

[0051] Such as Figure 5 As shown, MEAN can also inhibit the replication of mutant JC1 / N415D and JC1 / G451R viruses.

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Abstract

The invention relates to the technical field of medicines, and in particular relates to use of a compound MEAN represented in the formula (1) in the specification in preparation of a medicine for inhibiting HCV replication, wherein the medicine is a medicine for inhibiting genotype 2 HCV replication; the compound MEAN is combined with IFN-alpha / RBV / ITX 5061 so as to synergistically inhibit genotype 2 HCV replication; and MEAN is capable of inhibiting genotype 2 HCV replication.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the use of compound MEAN in the preparation of medicines for inhibiting HCV replication. Background technique [0002] Hepatitis C virus is a global infectious disease caused by hepatitis C virus (Hepatitis C virus, HCV) infection, and it is also one of the main causes of liver cirrhosis and liver cancer. About 80% of people infected with HCV will develop chronic hepatitis. Currently, pegylated interferon (PEG IFN) combined with ribavirin (RBV) is the standard treatment for chronic hepatitis C (CHC), but only about 50% of patients can achieve sustained virological response (SVR), And there are problems such as contraindications, individual differences and drug side effects. Therefore, there is an urgent need to develop new safe and effective alternative therapies. [0003] The successful establishment of HCV replicon in vitro culture model and infection model has played a hug...

Claims

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Application Information

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IPC IPC(8): A61K31/473A61P31/14
CPCA61K31/473
Inventor 薛继华陈智刘艳宁朱海红
Owner ZHEJIANG UNIV
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