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Application of synthetic peptide in preparation of medicine for preventing and treating hepatitis virus infection

A technology for hepatitis C virus and synthesizing peptides, applied in the field of biomedicine, can solve the problems of low virus response rate, large toxic and side effects, high price and the like, and achieve the effects of simple and convenient purification process, low production cost and high safety.

Active Publication Date: 2021-01-29
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The former treatment has severe side effects and low viral response rate, especially for patients with HCV I genotype (the main popular genotype in my country), the response rate is only 40-50%; DAAs mainly target NS3 / 4A protease and NS5A protease of HCV And NS5B polymerase, since the approval of the US Food and Drug Administration (FDA) in 2011, the overall clinical cure rate has reached about 90%, but with the increasing number of DAAs users, some problems of the drug have gradually begun to be exposed, such as , drug-resistant mutation, medication in patients with HBV / HCV co-infection will stimulate HBV replication (unfortunately, there are many hepatitis B infections in my country), there are interactions between DAAs, and the price is relatively expensive (so most HCV patients in my country are still Peg-IFN / RBV-based treatment plan), the adverse reactions of the drug itself are gradually appearing, including liver damage, kidney damage, skin damage, etc. In addition, whether the all-oral regimen can reduce the risk of hepatocellular carcinoma needs to be further investigated. Clearly wait for a series of questions

Method used

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  • Application of synthetic peptide in preparation of medicine for preventing and treating hepatitis virus infection
  • Application of synthetic peptide in preparation of medicine for preventing and treating hepatitis virus infection
  • Application of synthetic peptide in preparation of medicine for preventing and treating hepatitis virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Synthetic peptide inhibits the effective concentration screening experiment of HCV infection

[0028] 1.1 HCV infection experiment of Huh7.5.1 cells

[0029] Normal Huh7.5.1 cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Culture in an incubator with saturated humidity, and add 2mM L-glutamine, 0.1mM non-essential amino acids, 100μg / mL streptomycin and 100U / mL penicillin to the cell culture medium.

[0030] The day before infection, Huh7.5.1 cells were divided into 3 × 10 4 cells / well were seeded in a 96-well cell culture plate, placed at 37°C, 5% CO 2 Incubate overnight in an incubator. On the next day, first suck out the culture supernatant, rinse twice with pre-warmed PBS, inoculate HCV with a viral load of MOI=1, infect at 37°C for 5 hours, discard the virus solution, rinse three times with pre-warmed PBS, and replace with Fresh culture medium was cultured at 37°C for 48 hours, and the positive cells infected w...

Embodiment 2

[0045] Example 2: Cytotoxicity Test of Synthetic Peptides

[0046] The CCK-8 method was used to detect the effect of adding synthetic peptides on the proliferation of Huh7.5.1 cells, and the specific steps were as follows:

[0047] Collect Huh7.5.1 cells in the logarithmic growth phase at approximately 3×10 4 seeded in 96-well plates. After the cells grew overnight, the synthetic peptides of various concentrations in Example 1 were added, and the culture was continued for 48 hours, and then the cell proliferation was detected by the CCK-8 method. The specific detection method is as follows: discard the original medium in the cells, add 110 μL of fresh medium containing 10 μL CCK-8 to each well, place it in a 37°C incubator and continue to cultivate for 3 hours, and then use a multi-functional microplate reader to detect at a wavelength of 450nm The absorbance value of each well. The experiment was repeated three times independently, and the mean and standard error were calc...

Embodiment 3

[0049] Embodiment 3: the experiment that synthetic peptide suppresses HCV protein expression

[0050] 3.1 Inhibition experiment of synthetic peptides on HCV infection

[0051] The day before infection, Huh7.5.1 cells were divided into 2 × 10 5 cells / well were seeded in 24-well cell culture plates, placed at 37°C, 5% CO 2 Incubate overnight in an incubator. On the next day, the culture supernatant was first aspirated, rinsed twice with pre-warmed PBS, and HCV was inoculated at a viral load of MOI=1, in which 80 μM of the above-mentioned synthetic peptide QLP-98 was added, and after infecting the cells for 5 hours, the viral liquid was discarded. Replace with fresh culture medium and continue culturing for 48 hours, and detect the expression of virus protein by Western blotting.

[0052] 3.2 Western blotting to detect the expression of HCV Core protein

[0053] 3.2.1 Preparation of cell samples

[0054] After the cells in the control group and synthetic peptide treatment gr...

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Abstract

The present invention relates to the technical field of biomedical engineering, and provides a synthetic peptide having the activity of inhibiting hepatitis C virus infection and its use in the preparation of medicines for preventing or treating hepatitis C virus infection. The synthetic peptide of the present invention has such as SEQ ID The amino acid sequence shown in NO.1. It has been confirmed by experiments that the synthetic peptide can inhibit the infection of HCV to target cells, and can effectively block the expression and replication of viral proteins and genomic RNA in target cells, thereby confirming that the synthetic peptide of the present invention has the ability to effectively block the infection of HCV on the host. Infectious ability of cells. In addition, through cytotoxicity experiments, the synthetic peptides of various concentrations in the present invention will not have any impact on the normal physiological functions of cells, and are highly safe. Therefore, the present invention provides a new idea for the prevention and treatment of hepatitis C, and has potential good clinical application value.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a synthetic peptide having the activity of inhibiting hepatitis C virus infection, its potential medical application in the preparation of anti-hepatitis C drugs, and a drug using the synthetic peptide as an active ingredient. Background technique [0002] In recent years, the study of bioactive peptides has become one of the hot spots in the development of global medicine. It not only has the advantages of extensive sources, good safety, specific targeting and strong biological activity, but also has various physiological functions such as immune regulation, anti-virus, anti-tumor, anti-thrombosis, anti-oxidation, and cholesterol-lowering. It has become the current international drug It is a research object that is competing with the health care products industry, and its application development prospects are very promising. [0003] Since most bioactive peptides are isolat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00A61K38/16A61P1/16A61P31/14
CPCA61K38/00A61P1/16A61P31/14C07K14/001
Inventor 秦照玲戚中田刘彬
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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