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Therapeutic hepatitis b vaccine and preparation method and use thereof

A hepatitis B vaccine and therapeutic technology, applied in peptide preparation methods, chemical instruments and methods, pharmaceutical formulations, etc., can solve the problems of incomplete antiviral treatment and difficult removal, so as to avoid immune escape, strong antigenicity, and overcome less antigenic effect

Active Publication Date: 2009-02-11
广州市茵良强生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many difficulties and controversies in the implementation process
The persistence and replication of HBV is the main reason for the progressive development of the disease, so antiviral treatment is the key means, although the nucleoside (acid) analogues have made great progress, but due to the drug-resistant mutation of HBV, HBV DNA may be associated with Due to factors such as the integration of host genes and the difficulty of removing circular covalently closed DNA (cccDNA), antiviral therapy is not thorough

Method used

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  • Therapeutic hepatitis b vaccine and preparation method and use thereof
  • Therapeutic hepatitis b vaccine and preparation method and use thereof
  • Therapeutic hepatitis b vaccine and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Preparation of polypeptide epitope

[0039] The preparation of polypeptides uses the Fmoc solid-phase peptide synthesis method. First, an amino acid protected by an Fmoc group on the α-amino group is connected to an insoluble carrier through an arm, and then the α-amino group is deprotected, and the amino acid is washed with a solution- Arm - Resin. The second pre-activated α-amino protected amino acid is connected through a coupling reaction. After the condensation reaction is completed, it is washed with a solution, and the deprotection and coupling are repeated until the target peptide is obtained. Finally the peptide-arm-resin is cleaved. Peptide purification, using HPLC method, the crude peptide product is separated and purified by C18 high-pressure column, and the required effluent is tracked and collected by liquid chromatography, the sample peaks are combined, desalted, and freeze-dried to obtain a high-quality peptide. The following takes the prepar...

Embodiment 2

[0076] Example 2 Quality Control of Polypeptide Epitopes

[0077] Quality control testing items for peptide epitopes include: appearance, purity and molecular weight.

[0078] Appearance of the polypeptide: the appearance of the prepared epitope peptide should be white powder; chromatographic analysis of the polypeptide: use RP-HPLC to analyze the polypeptide, chromatographic conditions: Waters company C18 chromatographic column (250×4.6mm, 5μm, 10nm). Mobile phase A (acetonitrile with 0.1% trifluoroacetic acid). Mobile phase B (water with 0.1% trifluoroacetic acid). Linear gradient elution: 0.01 to 20.00min, B from 35% to 50%, flow rate: 1.0ml / min. Detection wavelength: 220nm. Injection volume: 10 μl. Column temperature: room temperature. Quality requirements: purity>98%; mass spectrometry identification of peptides: dissolve the sample into a 0.05 mg / ml solution with 30% acetonitrile aqueous solution. Take 0.51 points on the sample plate, and use 0.5 μl HCCA as the matri...

Embodiment 3

[0081] Example 3 Screening of polypeptide epitopes

[0082] At present, the design of hepatitis B therapeutic vaccine tends to cover most of the population. Only in this way can it be effective for most of the population and have better commercial value. Broad coverage can be achieved by selecting epitope peptides with high affinity to HLA supertypes that are highly expressed in the overall population.

[0083] HLA class I molecules can bind to multiple overlapping but independent peptides. According to the characteristics of the bound peptides, most HLA class I molecules can be classified into several major HLA class I supertypes. By examining the ability of these epitopes to bind to all or most HLA molecules of a specific supertype, the epitopes required for the preparation of effective multi-epitope vaccines can be screened. Selecting the population with the widest coverage and targeted selection of the most common HLA supertypes in some populations is conducive to the rat...

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Abstract

The invention discloses a curative hepatitis B vaccine and a preparation method and applications thereof. The key of the technical proposal is that: a plurality of CTL and HTL polypeptide epitopes originating from HBV envelope protein, core antigen and polymerase are selected to form a multiple antigenic peptide (MAP) system and recombinated human interleukin 12 (rhIL-12) is adopted as an adjuvant, so as to improve the immunogenicity and the response level, thus providing a biological preparation for curing chronic hepatitis B clinically.

Description

technical field [0001] The invention relates to a biological preparation, more specifically, it relates to a therapeutic hepatitis B vaccine for chronic hepatitis B, and the invention also relates to a preparation method and application of the vaccine. Background technique [0002] Chronic hepatitis B is one of the common infectious diseases in my country. my country is a high prevalence area of ​​hepatitis B virus (HBV). HBV infection is a serious public health problem, but no breakthrough has been made in the treatment of chronic hepatitis B. . [0003] Until now, the clinical treatment of chronic hepatitis B is still facing a lot of confusion, although the overall goal of treatment is clear, that is, to maximize the long-term suppression or elimination of HBV, reduce liver cell inflammation, necrosis and fibrosis, delay and prevent disease progression, etc. However, there are still many difficulties and disputes in the implementation process. The persistence and replica...

Claims

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Application Information

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IPC IPC(8): A61K38/20A61K38/08A61K38/10A61K47/42A61K47/36A61K47/32A61K47/26A61K47/10C07K7/06C07K7/08C07K1/04A61P31/20A61P35/00
Inventor 张宜俊邱向南夏书奇曾振飞叶倩君赵峰段详王艳王翠玲吴思纹于源李平黄观凤熊伟宏王增松
Owner 广州市茵良强生物科技有限公司
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