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A tuberculosis polypeptide vaccine for preventing and treating tuberculosis and its preparation method and application

A polypeptide vaccine and tuberculosis technology, which is applied to medical preparations without active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas, etc., can solve the problem of unknown tuberculosis vaccines, unknown anti-tuberculosis immune effects, and no tuberculosis antigen polypeptide tuberculosis. And other issues

Active Publication Date: 2021-08-17
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complexity and integrity of the molecular network regulation in the living body, it is still unknown whether these polypeptide fragments also play the same role in the body, so that they can be used to prepare tuberculosis vaccines
In addition, whether these polypeptide fragments are suitable for in vitro synthetic production, which polypeptide fragments can induce better levels of humoral immunity and cellular immunity in vivo, and which adjuvant (or carrier) should be used to achieve a better anti-tuberculosis immune effect is still unknown
However, there is no relevant research and report on chitosan and its derivatives as an adjuvant combined with tuberculosis antigen polypeptide for the prevention and treatment of tuberculosis.

Method used

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  • A tuberculosis polypeptide vaccine for preventing and treating tuberculosis and its preparation method and application
  • A tuberculosis polypeptide vaccine for preventing and treating tuberculosis and its preparation method and application
  • A tuberculosis polypeptide vaccine for preventing and treating tuberculosis and its preparation method and application

Examples

Experimental program
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Embodiment 1

[0045] In this embodiment, the tuberculosis polypeptide nano-vaccine using chitosan-deoxycholic acid conjugate as a carrier includes chitosan-deoxycholic acid conjugate and tuberculosis polypeptide; the tuberculosis polypeptide is composed of Mycobacterium tuberculosis Rv0180c protein 81 ~100 amino acid fragments, Rv0227c protein 261~280 amino acid fragments and Rv2004c protein 281~300 amino acid fragments.

[0046] The preparation of above-mentioned tuberculosis polypeptide nano-vaccine comprises the following steps:

[0047] (1) Synthesis of three tuberculosis polypeptides

[0048] The 81-100 amino acid fragment of the Mycobacterium tuberculosis Rv0180c protein, the 261-280 amino acid fragment of the Rv0227c protein and the 281-300 amino acid fragment of the Rv2004c protein were synthesized.

[0049] The basic physicochemical properties of amino acids 81-100 of Rv0180c protein are as follows: figure 1 Shown; The basic physicochemical properties of the 261-280 amino acids o...

Embodiment 2

[0061] In this embodiment, the tuberculosis polypeptide nano-vaccine with chitosan-deoxycholic acid conjugate as carrier includes chitosan-deoxycholic acid conjugate and tuberculosis polypeptide; the tuberculosis polypeptide is composed of Mycobacterium tuberculosis Rv0180c protein The 81-100 amino acid fragment and the 261-280 amino acid fragment of the Rv0227c protein consist of two polypeptides.

[0062] The preparation of above-mentioned tuberculosis polypeptide nano-vaccine comprises the following steps:

[0063] (1) Synthesis of two tuberculosis polypeptides

[0064] Synthetic method is the same as embodiment 1.

[0065] (2) Preparation of chitosan-deoxycholic acid conjugate

[0066] Dissolve 0.15 g of chitosan in 20 mL of 1% aqueous acetic acid, and then dilute with 20 mL of methanol. Add 0.8g of deoxycholic acid and 0.6g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCL) successively to the diluent, stir and react at room temperature 20h. Pou...

Embodiment 3

[0074] In this embodiment, the tuberculosis polypeptide nano-vaccine with chitosan-deoxycholic acid conjugate as a carrier includes chitosan-deoxycholic acid conjugate and tuberculosis polypeptide; the tuberculosis polypeptide is composed of Mycobacterium tuberculosis Rv0227c protein The 261-280 amino acid fragment and the 281-300 amino acid fragment of Rv2004c protein consist of two polypeptides.

[0075] The preparation of above-mentioned tuberculosis polypeptide nano-vaccine comprises the following steps:

[0076] (1) Synthesis of two tuberculosis polypeptides

[0077] Synthetic method is the same as embodiment 1.

[0078] (2) Preparation of chitosan-deoxycholic acid conjugate

[0079] Dissolve 0.25 g of chitosan in 20 mL of 1% aqueous acetic acid, and then dilute with 20 mL of methanol. Add 0.9g of deoxycholic acid and 0.65g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCL) successively to the diluent, stir and react at room temperature 28h. Pou...

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Abstract

The invention relates to a tuberculosis polypeptide vaccine for preventing and treating tuberculosis and its preparation method and application, belonging to the technical field of biopharmaceutical engineering. The tuberculosis polypeptide vaccine in the present invention includes a tuberculosis polypeptide and a carrier, and the tuberculosis polypeptide includes the 81-100 amino acid fragment of the Rv0180c protein of Mycobacterium tuberculosis, the 261-280 amino acid fragment of the Rv0227c protein, and the 281-300 amino acid fragment of the Rv2004c protein. one or more. The polypeptide fragments selected in the present invention are the key polypeptide fragments in the important protective antigens of MTB, and the main protective epitopes of multiple antigens are concentrated, which can be tightly combined with susceptible cells and effectively prevent Mycobacterium tuberculosis H37Rv from invading cells The natural structure and hydrophilicity of these three polypeptides are good, so they are suitable for artificial synthesis. Using chitosan-deoxycholic acid conjugate as a carrier can stimulate the body's effective humoral and cellular immune responses.

Description

technical field [0001] The invention relates to a tuberculosis polypeptide vaccine for preventing and treating tuberculosis and its preparation method and application, belonging to the technical field of biopharmaceutical engineering. Background technique [0002] Tuberculosis (TB) is a chronic wasting infectious disease caused by Mycobacterium tuberculosis complex (MTC). Mycobacterium tuberculosis complex mainly includes Mycobacterium tuberculosis (Mycobacterium tuberculosis), Mycobacterium bovis (Mycobacterium bovis), Mycobacterium africanum (Mycobacterium africanum), three mycobacteria with obvious pathogenicity to humans and animals. Worldwide, the morbidity and mortality of tuberculosis rank among the forefront of various infectious diseases. my country is one of the countries with a high burden of tuberculosis, and the epidemic situation is very severe. The number of tuberculosis patients accounts for about 1 / 4 of the world, and it is one of the diseases with the larg...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/04A61K39/39A61K9/51A61K47/36A61P31/06
CPCA61K9/5161A61K39/04A61K39/39A61K2039/55583A61K2039/57A61K2039/575A61K2039/6087
Inventor 薛云王梦娅高伟娜赵战勤祖向阳
Owner HENAN UNIV OF SCI & TECH
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