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Human endothelin type-A receptor immunogenic peptide fragment and vector vaccine thereof

An immunogenic, carrier vaccine technology, applied in hormone receptors, receptors/cell surface antigens/cell surface determinants, carrier-binding antigens/hapten components, etc. Effect

Active Publication Date: 2017-06-23
WUHAN HUAJIYUAN BIOTECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] How to select the appropriate target and carrier is the key to the successful development of therapeutic vaccines such as PAH, because therapeutic vaccines mainly target self-antigens or immune-tolerance foreign antigens, and therapeutic vaccines must be combined with carriers to break immune tolerance and produce therapeutic effects , selectable targets theoretically include endothelin 1 (Endothelin, ET-1) and human endothelin type A receptor (ET-1) A R), but due to the extremely low blood circulation concentration of ET-1, most of them act through paracrine and autocrine, and exert their effectiveness after ET-1 is produced before combining with ET-1 vaccine antibody. Therefore, ET-1 is not Ideal target for PAH therapy

Method used

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  • Human endothelin type-A receptor immunogenic peptide fragment and vector vaccine thereof
  • Human endothelin type-A receptor immunogenic peptide fragment and vector vaccine thereof
  • Human endothelin type-A receptor immunogenic peptide fragment and vector vaccine thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: ET A Preparation of R immunogenic peptides

[0024] According to the characteristics of bioinformatics technology, such as amino acid hydrophilicity, spatial conformation, and B cell epitope, the ET A ET of R extracellular amino acid sequence A 9 R immunogenic peptides, respectively named HI10, RF9, EC10, YC9, RC8, EM7, TF10, CK8, NE9, the specific amino acid sequences are, HI10: SEQ ID No.1; RF9: SEQ ID No.2 ; EC10: SEQ ID No.3; YC9: SEQ ID No.4; RC8: SEQ ID No.5; EM7: SEQ ID No.6; TF10: SEQ ID No.7; CK8: SEQ ID No.8, NE9: SEQ ID No. ID No. 9.

[0025] The above-mentioned 9 peptides were synthesized by PSSM-8 automatic peptide synthesizer (SHIMADZU company, Japan), and the purity of the 9 peptides was analyzed by high performance liquid chromatography, and the purity of the 9 peptides was tested. Reach more than 92%. The obtained 9 peptides were freeze-dried, aliquoted, placed in cryopreservation tubes, and stored at -80°C for later use.

Embodiment 2

[0026] Example 2: Preparation of ET A R immunogenic vector vaccine

[0027] 1. Using Qβ-2aa bacteriophage virus-like particle protein to prepare 9 kinds of vector vaccine ET A R RF9-Qβ, ET A REC10-Qβ, ET A R YC9-Qβ, ET A R RC8-Qβ, ET A R EM7-Qβ, ET A R TF10-Qβ, ET A R CK8-Qβ, ET A R HI10-Qβ, ET A RNE9-Qβ. Concrete preparation process is as follows:

[0028] 1) Preparation of Qβ-2aa bacteriophage VLP: the English abbreviation of Qβ-2aa phage VLP is: Qβ-2aa VLP, hereinafter Qβ-2aa VLP is used to represent Qβ-2aa phage VLP.

[0029] The preparation method of Qβ-2aa VLP is:

[0030] 1a) Obtaining a recombinant strain expressing Qβ-2aa VLP: the recombinant strain is Escherichia coli DH5α / pGEXQβ-A1, and this recombinant strain can induce the production of Qβ-2aa VLP protein. The preservation number of Escherichia coli DH5α / pGEXQβ-A1 is CCTCCNO: M209282. For the specific preparation process, please refer to the Chinese patent: A Qβ-2aa bacteriophage virus-like particle pr...

Embodiment 3

[0044] Example 3ET A In Vitro Functional Study of Antibodies Targeted by R Immunogenic Vector Vaccines

[0045] 1. ET A Screening of R immunogenic peptides: ET is preferred A R immunogenic peptides are EC10, YC9, RC8, EM7, TF10. The specific test process is:

[0046] 9 kinds of ET that embodiment 2 obtains A R immunogenic vector vaccine: ET A R RF9-Qβ, ET A R EC10-Qβ, ET A RYC9-Qβ, ET A R RC8-Qβ, ET A R EM7-Qβ, ET A R TF10-Qβ, ET A R CK8-Qβ, ET A R HI10-Qβ, ET A R NE9-Qβ subcutaneously immunized male New Zealand white rabbits at multiple points on days 1, 14, and 28. Among them, as a control group, 2 male New Zealand white rabbits were immunized with vector Qβ-2aa-CPG-ODN VLP, and each ET A R immunogenic carrier vaccine immunized 2 male New Zealand white rabbits, a total of 20 immunized male New Zealand white rabbits, and the immunization dose was 500ug per rabbit. On the 7th, 14th, 21st, 28th, and 35th days, the blood was taken to measure the antibody titer by E...

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Abstract

The invention belongs to the technical field of biology, and discloses a human endothelin type-A receptor immunogenic peptide fragment and a vector vaccine thereof. The human endothelin type-A receptor immunogenic peptide fragment has one amino acid sequence selected from the group consisting of SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 7, SEQ ID No. 8 and SEQ ID No. 9. The human endothelin type-A receptor immunogenic vector vaccine is prepared from the human endothelin type-A receptor immunogenic peptide fragment and a vector by coupling, wherein the vector is Q beta-2aa phage virus-like particle protein or keyhole limpet haemocyanin. The human endothelin type-A receptor immunogenic vector vaccine obtained in the invention can effectively treat pulmonary arterial hypertension.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a human endothelin type A receptor immunogenic peptide segment and a carrier vaccine thereof. Background technique [0002] Pulmonary arterial hypertension (PAH) is a severe chronic disease characterized by the hyperplasia and remodeling of pulmonary arterioles, leading to a progressive increase in pulmonary artery pressure and eventually right heart failure and death. Its incidence is relatively low, but its prognosis is extremely poor. The 5-year survival rate was 20.8%. In the past 20 years, drugs for the treatment of PAH have achieved certain curative effects, such as endothelin receptor antagonists, which prolong the median survival time of PAH from 2.8 years in the 1980s to 5 years, but these drugs are expensive and difficult to popularize in treatment. There is an urgent need to find new treatments that are convenient, effective and inexpensive. Therapeutic vacci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/72A61K39/385A61K39/00A61P11/00A61P9/12
CPCA61K39/008A61K39/385A61K2039/6081C07K14/72
Inventor 廖玉华宋霄霄周子华邱志华陈霄
Owner WUHAN HUAJIYUAN BIOTECH DEV
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