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Short peptide for inhibiting human hepatitis B virus infection and its application

A hepatitis B virus, short peptide technology, applied in the direction of viral peptides, applications, antiviral agents, etc., can solve the problems of drug cross-resistance, single target, etc., to achieve good specificity, high inhibition efficiency, good application Foreground effect

Active Publication Date: 2012-11-14
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the single target of the existing antiviral drugs, once the hepatitis B virus develops drug resistance to a certain drug, it is very easy to cause cross-resistance to other drugs

Method used

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  • Short peptide for inhibiting human hepatitis B virus infection and its application
  • Short peptide for inhibiting human hepatitis B virus infection and its application
  • Short peptide for inhibiting human hepatitis B virus infection and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Short peptides are able to bind preS1 13-59 .

[0043] Biotin-coupled short peptides were bound to Streptavidin-coated 96-well plates, 50ug Biotin-coupled short peptides were added to each well, and combined in 100ul PBS at 4°C for 4 hours. Gently tap off the PBS, rinse with PBS again. Re-add 100ul PBS and add 50ug myristoylated preS1 13-59 , 4°C for 8-12 hours. Gently tap off the PBS, rinse with PBS again. Add 200ul of PBS containing 0.1% BSA, combine at 37°C for 1 hour, wash once with PBST, 5 seconds each time. Then add 100ul anti-preS1 monoclonal antibody (1:1000, diluted in PBS containing 0.1% BSA), bind at 4°C for 4 hours, and then wash three times with PBST, 5 seconds each time. Add 100ul rabbit anti-mouse secondary antibody (1:1000, diluted in PBS containing 0.1% BSA), combine for 2 hours at 4 degrees, and then wash three times with PBST, 5 seconds each time. Finally, 100ul of TMB (premixed hydrogen peroxide) was added to each well, and the color...

Embodiment 2

[0046] Example 2: Short peptides can inhibit the infection of hepatitis B virus in tree shrew primary liver cell model

[0047] 2.1 Culture of tree shrew primary hepatocytes

[0048] Tree shrews were purchased from the Experimental Animal Center of Kunming Institute of Zoology, Chinese Academy of Sciences. Tree shrew primary hepatocytes were isolated by a two-step perfusion method. Select adult tree shrews with a weight of 160-200 grams. After feeding for 5-7 days, fast for 12 hours before operation. Anesthesia: inject ketamine hydrochloride 5mg / 100g body weight into each muscle, add xylazine 1mg / 100g body weight. Perfusion: Fix the tree shrew on the support, open the abdominal cavity, expose the portal vein, insert the catheter, and start perfusing Hanks solution preheated at 42°C (5mM EGTA, 1% glucose, 50 u / ml ampicillin, 100 μg / ml streptomycin, 100mM L-glutamine, 15mM HEPES, pH7.4), 20 ml / min, perfuse for 5 minutes. Hanks solution (Hanks solution Add 0.5 mg / ml t...

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Abstract

The invention belongs to the biomedicine field, and relates to a short peptide for inhibiting human hepatitis B virus infection. The short peptide comprises anyone of the following amino acid sequences: 1, N-leucine-arginine-asparagine-isoleucine-arginine-C; 2, an amino acid sequence obtained through deleting one or two amino acids from the above amino acid sequence 1 and capable of inhibiting the human hepatitis B virus infection; 3, an amino acid sequence obtained through substituting one or two amino acids in the amino acid sequence 1 and capable of inhibiting the human hepatitis B virus infection; 4, adding one or more amino acids to the amino acid sequence 1 and capable of inhibiting the human hepatitis B virus infection; and 5, an amino acid sequence obtained through repeated splicing of the amino acid sequences 1-4 and capable of inhibiting the human hepatitis B virus infection. The invention also provides a construct and a cell of an amino acid sequence comprising the short peptide or of a nucleic acid coding sequence of the short peptide, and an application of the short peptide.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a short peptide capable of inhibiting human hepatitis B virus from infecting cells, and the application of the short peptide in preparing medicines for treating human hepatitis B virus infection or hepatitis B. Background technique [0002] China is a high-risk area for human hepatitis B virus (abbreviation: hepatitis B virus) infection, and more than 8% of the population are carriers of hepatitis B virus. Chronic hepatitis B caused by hepatitis B virus infection is prone to develop into liver cirrhosis and liver cancer. Currently, only interferon and nucleoside (acid) analogues are available for the treatment of chronic hepatitis B. The response rate of the former is only about 30%, and the side effects are large; the latter is a hepatitis B virus polymerase inhibitor, and patients must receive long-term antiviral drug treatment, and severe drug resistance often occurs. Due to the si...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K7/08C07K14/02C12N15/51C12N15/63C12N1/21C12N5/10A61K39/29A61P31/20A61P1/16
Inventor 谢幼华何永刚汪垣邓强刘晶周佳海董琦蕾
Owner FUDAN UNIV
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