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Polypeptide medicine against hepatitis B virus X protein

A hepatitis B virus, anti-hepatitis B technology, applied in the direction of viral peptides, antiviral agents, viruses, etc., can solve the problems of short half-life of polypeptide fragments, affecting pharmacodynamics, and difficulty in discovering polypeptide drugs, and achieve obvious efficacy. effect of learning

Active Publication Date: 2014-02-05
TIANJIN TOPTECH BIO SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the discovery of peptide drugs is difficult because most of the peptide fragments have a short half-life in the body, which directly affects the pharmacodynamic effect.

Method used

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  • Polypeptide medicine against hepatitis B virus X protein
  • Polypeptide medicine against hepatitis B virus X protein
  • Polypeptide medicine against hepatitis B virus X protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1: Design and preparation of polypeptides

[0101] Synthetic polypeptide functional fragment anti-HBxP2#:

[0102] The present invention has synthesized the polypeptide (hereinafter referred to as Anti -HBxP2#). The preparation of the polypeptide adopts a solid-phase synthesis method, such as using an AAPPTECAPex396 polypeptide synthesis instrument (purchased from Hong Kong Universal Analytical and Testing Instrument Co., Ltd.), in a closed explosion-proof glass reactor to make amino acids according to the sequence shown in SEQ ID NO: 1, From C-terminus-carboxyl-terminus to N-terminus-amino-terminus, this refers to the first one added to the amino acid sequence Pro-Asp-Leu-His-Lys-Asn-Glu-Leu-Lys-His-Val-Lys-Tyr The amino acid monomer is Tyr at the C-terminus, then Lys, then Val, until the last Asp and Pro at the N-terminus, which are continuously added, reacted, synthesized, and operated to finally obtain the desired amino acid sequence. The solid-phase synt...

Embodiment 2

[0109] Embodiment two: the anti-HBx activity of isolated (in vitro) polypeptide

[0110] Two methods are used to detect the anti-HBx activity of the polypeptide in Example 1 in vitro: the first method is to use molecular cloning technology to clone the cDNA expressing the polypeptide in Example 1 into the eukaryotic expression vector pcDNA3.1 On (+), through gene transfection, the purpose of expressing the studied polypeptide in liver cancer cells is realized, and then the effect of the researched polypeptide on inhibiting HBx is observed; the second method is to use artificially synthesized polypeptides and directly add them to cultured liver cancer cells In the culture medium, observe the effect of polypeptide on inhibiting HBx.

[0111] There are two kinds of liver cancer cells used in the experiment: one is liver cancer HepG2-X cells constitutively expressing HBx (hepatoma HepG2 cells stably transfected with HBx); the other is liver cancer HepG2.2.15 cells constitutively e...

Embodiment 3

[0190] Example 3: Experiments on the effectiveness of polypeptides in vivo

[0191] HepG2-X cells or HepG2.2.15 cells in the logarithmic growth phase were digested with trypsin to make a cell suspension, the number of cells was calculated, and the cells were diluted to 1×10 with sterile normal saline. 7 cells / ml and stored in ice water. Twelve 4- to 6-week-old female BALB / C nude mice were then randomly divided into 2 groups: ① Control group, subcutaneously inject 0.2ml of the above-mentioned diluted cells in the armpit of the right forelimb of each mouse, and only inject 0.5 ml sterilized distilled water (without polypeptide drugs); ② experimental group (administration dose is 10 mg / kg body weight). Subcutaneously inject 0.2ml of the above-mentioned diluted cells into the armpit of the right forelimb of each mouse, and after 7 days of injection, the tumor volume (V=L×W 2 ×0.5) up to 100mm 3 , and then subcutaneously inject the above polypeptide drugs (dissolve the freeze-dr...

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Abstract

Disclosed are a polypeptide having the functional activity of inhibiting hepatitis B virus x protein (HBx) and a polynucleotide encoding the peptide. Also provided are peptidomimetics of the peptide, comprising functional fragments and functional variants, as well as encoding genes thereof. The peptide and peptidomimetics can be used for the prevention and treatment of liver diseases caused by hepatitis B virus infection, including hepatitis, cirrhosis and liver cancer.

Description

technical field [0001] The invention relates to the field of polypeptide medicine, in particular to a polypeptide against hepatitis B virus X protein, a polynucleotide encoding the polypeptide, and their application. Background technique [0002] Liver cancer is one of the malignant tumors that lead to the death of patients. Its malignancy is high. In my country, the death rate of liver cancer is second only to gastric cancer, ranking second in the death rate of malignant tumors in my country. According to statistics, there are 300,000 new cases of liver cancer each year in my country, and 110,000 deaths from liver cancer each year. Hepatitis B virus (HBV) infection can lead to hepatitis, liver cirrhosis, and primary liver cancer. In my country, more than 80% of liver cancer patients are HCC after HBV infection, which can be called post-HBV HCC. [0003] HBV is a DNA virus with a length of about 3.2Kb, and its open reading frame expresses hepatitis B virus surface antigen (...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C12N15/11C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A61K38/10A61K48/00A61P1/16A61P31/20
CPCC07K14/02C07K7/08C07K14/00A61K38/00A61P1/16A61P31/20A61P35/00C07K14/005C12N2730/10122
Inventor 张晓东叶丽虹
Owner TIANJIN TOPTECH BIO SCI & TECH
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