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Application of a kind of human β-defensin-1 in the preparation of medicine for treating or preventing hepatitis B virus infection

A technology of hepatitis B virus and defensin, which can be applied to antiviral agents, pharmaceutical formulas, medical preparations containing active ingredients, etc., and can solve problems such as unclear specific mechanisms

Active Publication Date: 2018-02-13
广东龙帆生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The only research on the effect of β-defensin-1 on viruses is its anti-HIV activity, and its specific mechanism is still unclear

Method used

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  • Application of a kind of human β-defensin-1 in the preparation of medicine for treating or preventing hepatitis B virus infection
  • Application of a kind of human β-defensin-1 in the preparation of medicine for treating or preventing hepatitis B virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Evaluation of anti-HBV activity of human β-defensin-1 by overexpression experiment.

[0028] A kind of application of human β-defensin-1 in the preparation of medicine for treating or preventing hepatitis B virus infection, the steps are:

[0029] 1. Experimental materials:

[0030] 1.1 Cells and plasmids:

[0031] 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 hBD1 gene fragment with specific primers. The primer sequences are as follows:

[0032] hBD1CDS Forward: 5'-CGCGAATTCGATGAGAACTTCCTACCTTC-3'

[0033] hBD1CDS Reverse:5'-TATCTCGAGCTTGCAGCACTTGGCCTTC-3'

[0034] The hBD1 gene was cloned into the vector pCAGGS (purchased from addgene) by specific restriction endonuclease (EcoR I and Xho I) digestion and ligation to obtain the overexpression plasmid of pCAGGS-hBD1, which was ide...

Embodiment 2

[0043] Example 2: Evaluation of the anti-HBV activity of human β-defensin-1 by interference experiments.

[0044] A kind of application of human β-defensin-1 in the preparation of medicine for treating or preventing hepatitis B virus infection, the steps are:

[0045] 1. Experimental materials:

[0046] The interference RNA package was purchased from Guangzhou Ruibo Company.

[0047] 2. The experimental method is the same as in Example 1.

[0048] 3. Experimental results:

[0049] Firstly, the interference effect of interfering RNA was detected, such as figure 1 As shown in C, the inhibition efficiency is about 80%. like figure 1 D, Interference hBD1 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 hBD1 inhibits HBV replication and hBD1 has anti-HBV activity.

[0050] Other steps are the same as in Example 1.

[0051] The experimental results of the above examples ...

Embodiment 3

[0052] Example 3 MTS detects the effect of transfection pCAGGS-hBD1 on cell viability

[0053] like figure 2 The pCAGGS-hBD1 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.

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Abstract

The invention discloses the application of β-defensin-1 in the preparation of medicines for treating or preventing hepatitis B virus infection. Study on the effect of β-defensin-1 on hepatitis B virus by co-transfection of β-defensin-1 overexpression plasmid or small interfering RNA targeting β-defensin-1 and HBV1.3 ploidy plasmid into liver cells Effects on replication, the surface antigen and e-antigen secreted by the virus were examined, as well as the levels of intracellular viral nucleocapsid-associated DNA, showing that β‑defensin‑1 inhibits HBV replication in hepatocytes. β-defensin-1 may affect the activity of HBV or affect the replication process of HBV infection or after infection. Therefore, by using alone or in combination with other anti-HBV drugs, β-defensin-1 can play an important role in the prevention or treatment of hepatitis B virus infection. Human β‑defensin‑1 is an antimicrobial polypeptide constitutively expressed in the human body, and has no toxic side effects on the human body at physiological concentrations.

Description

technical field [0001] The present invention relates to the field of antiviral drugs, relates to a new application of human β-defensin-1 (β-defensin-1, hBD1), more specifically relates to a kind of human β-defensin-1 in the preparation of treatment or prevention of hepatitis B The application of hepatitis virus (Hepatitis Bvirus, HBV) infection medicine. 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. The surface of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P31/20
Inventor 吴建国邬开朗谢雪萍陈俊波谭秋萍刘芳
Owner 广东龙帆生物科技有限公司
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