Peptide segment 502-518 of the highly conserved region of hcv envelope protein and its use

A highly conserved, envelope protein technology, applied in the field of biomedical engineering, can solve the problems of unconserved amino acid sequence, induce high-titer antibodies, and cannot be used to treat or prevent hepatitis C, and achieve the effect of preventing infection

Active Publication Date: 2021-06-29
GUANGZHOU EIGHTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these studies have not verified whether the above epitope peptides can induce high-titer antibodies in vivo, or whether the antibodies have a preventive effect on genotype 1-6 HCV infection in infection models.
[0006] Chinese patent CN102227445B discloses an antibody that binds to the envelope protein of hepatitis C virus and a method for identifying the genotype of hepatitis C virus using the antibody. The antibody disclosed in this invention specifically binds to the envelope of HCV of genotype 2a protein, but does not immunoreact with the envelope protein of HCV genotype 1a, resulting in the amino acid sequence corresponding to the antibody is not conserved in HCV virus; moreover, the antibody can only be used to alleviate the hepatitis C caused by HCV Adverse reactions, not for the treatment or prevention of hepatitis C caused by HCV

Method used

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  • Peptide segment 502-518 of the highly conserved region of hcv envelope protein and its use
  • Peptide segment 502-518 of the highly conserved region of hcv envelope protein and its use
  • Peptide segment 502-518 of the highly conserved region of hcv envelope protein and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 A pharmaceutical composition containing HCV highly conserved region peptides 502-518

[0044] The pharmaceutical composition containing highly conserved region peptides 502-518 of HCV includes peptides 502-518 of the highly conserved region on the HCV envelope protein, the pharmaceutical carrier is water for injection, the pharmaceutical excipient lubricant simethicone, Cross-linking agent sodium alginate, binder ethyl cellulose, preservative ethylparaben, pharmaceutically acceptable barbiturate.

[0045] The preparation method of the 1-6 type HCV pharmaceutical composition is:

[0046] a. Obtain 536 sequences covering genotype 1-6 HCV from the NCBI website (see literature: Deng K.etal..PloS one 10, 2015, e0138756) and compare them to obtain highly conserved on the HCV envelope protein The peptide segment 502-518 in the region, and the peptide segment was synthesized at Shanghai Gil Biochemical Company to obtain the amino acid peptide segment; the purity of t...

Embodiment 2

[0050] Embodiment 2 A kind of vaccine for prevention and treatment of hepatitis C

[0051] The preventive and therapeutic hepatitis C vaccine includes the coupling protein obtained by coupling the peptide segment 502-518 of the highly conserved region of the envelope protein with KLH, Freund's adjuvant, preservative thimerosal, inactivator formaldehyde, and stabilizer lactose and phosphate buffered saline.

[0052] The preparation method of the preventive and therapeutic hepatitis C vaccine is:

[0053] ①Coupling the peptide segment 502-518 of the highly conserved region of the HCV envelope protein with KLH to obtain a coupled protein; the coupling process of the peptide segment and KLH is in accordance with the manufacturer's instructions of the KLH-peptide conjugation kits kit from CellMosaic operating;

[0054] ②Introducing the coupling protein obtained in step ① into isolated chimpanzee hepatocytes to obtain recombined chimpanzee hepatocytes;

[0055] ③ adding inactivat...

Embodiment 3

[0058] The immunization of embodiment 3 Balb / c mice

[0059] A total of 536 HCV sequences of 1-6 genotypes were downloaded from the NCBI (https: / / www.ncbi.nlm.nih.gov / ) website, through the website https: / / www.ebi.ac.uk / Tools / msa / Sequence analysis on clustalo / obtained peptides 502-518 of the highly conserved region of the HCV envelope protein; at the same time, since 685-693 is located in the transmembrane region of the envelope protein and is not exposed, neutralizing antibodies will not be produced theoretically. It is used as a negative control peptide in the experiment of the present invention.

[0060] The peptides 502-518 and the negative control peptides 685-693 were synthesized in Shanghai Gil Biochemical Co., Ltd. with a purity of over 99%, and were coupled with KLH as immune antigens.

[0061] Two peptide immunization antigens were used to immunize six 8-10-week-old Balb / c mice, and six unimmunized negative control mice were set up. The specific immunization proce...

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Abstract

The invention belongs to the technical field of biomedical engineering, and in particular relates to a peptide segment 502-518 in a highly conserved region of HCV envelope protein and its application. The peptide segment of the highly conserved region of the HCV envelope protein provided by the present invention is a peptide segment 502 of a highly conserved region of the HCV envelope protein provided by sequence comparison of the highly conserved region of the HCV envelope protein and combining the spatial structure of the envelope protein ‑518, the amino acid sequence is Val‑Cys‑Gly‑Pro‑Val‑Tyr‑Cys‑Phe‑Thr‑Pro‑Ser‑Pro‑Val‑Val‑Val‑Gly‑Thr, which has been verified by animal experiments that this peptide can Induce highly efficient antibodies with broad-spectrum neutralizing effect on genotype 1-6 HCV. The highly conserved peptide segment 502-518 derived from the HCV envelope protein provided by the present invention can also be used to prepare vaccines for preventing and treating hepatitis C, and has the advantages of high conservation and wide application range.

Description

technical field [0001] The invention belongs to the technical field of biomedical engineering, and specifically relates to a highly conserved peptide segment 502-518 derived from HCV envelope protein and its application. Background technique [0002] Hepatitis C virus (HCV) is a single-stranded positive-sense RNA virus belonging to the family Flaviviridae. About 3% of the world's population is infected with HCV, of which 75%-85% develop chronic hepatitis and are at risk of developing cirrhosis and liver cancer. In addition, HCV has very high gene variability, and HCV can be divided into 1-7 genotypes according to the differences in genome phylogeny and sequence. The dominant genotypes in different countries and regions are not the same, and the 1b and 2a genotypes are more common in my country. Due to the successful development of direct acting antivirals (DAAs), the treatment of hepatitis C has achieved good results. However, there is still a lack of vaccines to prevent ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/18A61K39/12A61P31/14
CPCA61K39/12A61P31/14C07K14/005C12N2770/24222C12N2770/24234
Inventor 邓凯
Owner GUANGZHOU EIGHTH PEOPLES HOSPITAL
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