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Application of a kind of human homeobox protein a10 in the preparation of medicine for treating or preventing hepatitis B virus infection

A technology of hepatitis B virus and homology box, which is applied in the direction of drug combination, antiviral agent, peptide/protein composition, etc.

Active Publication Date: 2020-07-10
广东龙帆生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, no studies have reported the relationship between HOXA10 expression and HBV replication

Method used

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  • Application of a kind of human homeobox protein a10 in the preparation of medicine for treating or preventing hepatitis B virus infection
  • Application of a kind of human homeobox protein a10 in the preparation of medicine for treating or preventing hepatitis B virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028]Example 1: Evaluation of anti-HBV activity of human homeobox protein A10 by overexpression experiment.

[0029] A kind of application of human homeobox protein A10 in the preparation treatment or prevention medicine of hepatitis B virus infection, its steps are:

[0030] 1. Experimental materials:

[0031] 1.1 Cells and plasmids:

[0032] HepG2 cells (from ATCC) and pHBV1.3 plasmid. Using the total mRNA extracted from Huh7 cells as a template, first amplify the total cDNA by reverse transcription PCR, and then use the cDNA as a template to amplify the complete HOXA10 gene fragment with specific primers. The primer sequences are as follows:

[0033] HOXA10CDS Forward: 5'-CGCGAATTCGATGAGAACTTCCTACCTTC-3';

[0034] HOXA10 CDS Reverse: 5'-TATCTCGAGCTTGCAGCACTTGGCCTTC-3'.

[0035] The HOXA10 gene was cloned into the vector pCAGGS (purchased from addgene) by specific restriction endonucleases (EcoRI and XhoI) and ligated to obtain an overexpression plasmid of pCAGGS-HOXA10...

Embodiment 2

[0046] Example 2: Evaluation of anti-HBV activity of human homeobox protein A10 by interference experiment.

[0047] A kind of application of human homeobox protein A10 in the preparation treatment or prevention medicine of hepatitis B virus infection, its steps are:

[0048] 1. Experimental materials:

[0049] Interfering RNA package (purchased from Guangzhou Ruibo Company).

[0050] 2. Experimental method

[0051] Same as Example 1.

[0052] 3. Experimental results:

[0053] Firstly, the interference effect of interfering RNA was detected, such as figure 1 As shown in C, the inhibition efficiency is about 80%. Such as figure 1 D, Interference HOXA10 shown in 1E up-regulates the expression of HBeAg and HBsAg and the level of HBV replication intermediate nucleocapsid-related DNA in cells, indicating that HOXA10 inhibits HBV replication and HOXA10 has anti-HBV activity.

[0054] Other steps are the same as in Example 1.

[0055] The experimental results of the above exa...

Embodiment 3

[0056] Example 3: MTS detects the effect of transfection pCAGGS-HOXA10 on cell viability

[0057] A kind of application of human homeobox protein A10 in the preparation treatment or prevention medicine of hepatitis B virus infection, its steps are:

[0058] 1. Experimental materials:

[0059] Interfering RNA package (purchased from Guangzhou Ruibo Company).

[0060] 2. Experimental method.

[0061] Such as figure 2 The pCAGGS-HOXA10 plasmid corresponding to the amount of transfection shown in the HepG2 cells was used as a control with pCAGGS empty load, and the cell viability was detected with MTS (purchased from Promega) 48 hours after transfection. HepG2 cell viability was not affected.

[0062] Other implementation steps are the same as in Example 1.

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Abstract

The invention discloses application of human HOXA10 (Homeobox A10) in preparing a drug for treating or preventing HBV (Hepatitis B Virus) infection. The influence of HOXA10 on HBV replication can be researched through a way of cotransferring HOXA10 overexpression plasmids or small interfering RNA (Ribonucleic Acid) and HBV 1.3-ploid plasmids of targeted HOXA10 to hepatic cells, a surface antigen and an e antigen secreted by viruses and the level of DNA (Deoxyribonucleic Acid) related to virus nucleocapsid in cells are detected, and the HBV replication in hepatic cell inhibition by the HOXA10 is displayed; the HOXA10 can affect the activity of HBV or affect the HBV infection or a replication process after infection. Therefore, through independent use or combined use with other anti-HBV drugs, the HOXA10 can play an important role during a process of preventing or treating the HBV infection; the HOXA10 is a cell factor which is constitutively expressed in a human body, and the HOXA10 hasno toxic and side effects on the human body under physiological concentration.

Description

technical field [0001] The present invention relates to the field of antiviral drugs, relates to a new application of human homeobox A10 (homeobox A10, HOXA10), more specifically relates to a kind of human homeobox A10 in the preparation of treatment or prevention of hepatitis B virus (HepatitisB virus , HBV) infection drug application. Background technique [0002] Hepatitis B virus (HBV) infection is one of the main factors causing liver cancer, which seriously threatens human life and health. my country is a high-incidence area of ​​hepatitis B virus infection. There are about 120 million chronic hepatitis B patients, and about 300,000 people die of hepatocellular carcinoma caused by chronic hepatitis B every year. HBV belongs to the hepadnaviridae family, and its genome is partially double-stranded circular DNA. The core particle diameter of HBV is 28nm, which is a 20-hedron, and the viral capsid is composed of the core protein HBc. There is an envelope on the surface...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P31/20A61P1/16
CPCA61K38/17A61P1/16A61P31/20
Inventor 吴建国邬开朗杨庆雨刘映乐谭秋萍
Owner 广东龙帆生物科技有限公司
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