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Angiotensin II receptor 1 type polypeptide-vector vaccine and application thereof

An angiotensin, carrier vaccine technology, applied in cardiovascular system diseases, antibody medical ingredients, medical preparations with non-active ingredients, etc. Poor treatment compliance and other problems, to achieve good target organ protection and lower blood pressure

Active Publication Date: 2011-11-23
WUHAN HUAJIYUAN BIOTECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned renin inhibitors, ACEI and ARB all need to be taken daily, and it is long-term or even life-long. The overall economic burden is heavy, and the adverse reactions of the drugs themselves lead to poor treatment compliance of hypertensive patients. And the target organ protection effect also failed to achieve the maximum effect

Method used

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  • Angiotensin II receptor 1 type polypeptide-vector vaccine and application thereof
  • Angiotensin II receptor 1 type polypeptide-vector vaccine and application thereof
  • Angiotensin II receptor 1 type polypeptide-vector vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0033] Implementation Case 1 Preparation and Identification of ATRQβ-001 Vaccine

[0034] The short peptide CGGAFHYESQ is directly synthesized by Shanghai Gil Biochemical Co., Ltd., and the purity of the short peptide is 95%.

[0035] The vector Qβ-2aa virus-like particle was constructed, expressed and purified by our laboratory, with a purity of more than 90%. It was prepared according to the following method:

[0036] 1) After the stop codon of Qβ bacteriophage CP protein gene was mutated from TGA to strong stop codon TAA, it was cloned into the prokaryotic expression vector pET28a(+), and the CP protein particle pETQβ-CP was obtained;

[0037] 2) In the Qβ phage CP elongation protein gene encoding the position of the immunodominance determining region, that is, between the 72nd and 73rd codons of the Qβ bacteriophage CP elongation protein gene, insert the nucleotide AAGCTT encoding lysine and leucine, and The stop codon of the CP protein gene was mutated from TGA to GGA an...

Embodiment example 2

[0048] Implementation Case 2 Anti-ATRQβ-001 Vaccine Short Peptide Antibody Titer

[0049] Male spontaneously hypertensive rats came from the Shanghai Slack Experimental Animal Center, and the animals were raised in the SPF Animal Experiment Center of Tongji Medical College, using 12h / 12h light and non-restricted ordinary diet. Specifically divided into 4 groups: ATRQβ-001 vaccine immunization group; Ang II-Qβ vaccine immunization group (Ang II derivative and Qβ-2aa virus-like particle coupled vaccine); valsartan drug (Novartis, Switzerland) gavage group; PBS control SHR group. Except for the 8-week-old valsartan drug administration group, the other groups were all 6-week-old male SHR rats, with 9 rats in each group.

[0050] Take an equal volume of vaccine (not enough to make up with PBS) and PBS, inject subcutaneously at 3-4 points on the back of the rats, 400ul / rat, the vaccine dose of the vaccine group is about 300ug / rat, and boost twice every two weeks after the initial i...

Embodiment example 3

[0051] Implementation Case 3 Specificity of Anti-ATRQβ-001 Vaccine Short Peptide Antibody

[0052] The anti-peptide antibody CQ-8 of ATRQβ-001 vaccine was purified, and the specificity of ATRQβ-001 anti-peptide antibody CQ-8 was identified by immunoblotting and cellular immunofluorescence. Three groups were set up: CQ-8 was the anti-peptide antibody of animals immunized with ATRQβ-001 vaccine, DCQ-8 was the mixture after CQ-8 was neutralized by ATR-001 short peptide, and CON was the negative control antibody. Extract SD rat mesenteric tertiary arterial smooth muscle cell total protein, carry out immunoblotting analysis, the result shows that CQ-8 can combine with the protein of about 41kDa size in this total protein extract, and this protein has the same molecular weight as rat AT1A receptor (approximately 41kDa) consistent ( image 3 A). At the same time, it was found by cellular immunofluorescence that CQ-8 can bind to the AT1A receptor on the surface of rat mesenteric ter...

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Abstract

The invention discloses an angiotensin II receptor 1 type polypeptide-vector vaccine and an application thereof, belonging to the field of biotechnology drugs and biologic therapeutics. An angiotensin II receptor 1 type immunogenic peptide containing a first connection locus, a derivant thereof and one or more of carriers are preferably coupled and connected with recombined Qbeta-2aa bacteriophage viruslike particle protein to form an ordered and repeated polypeptide-carrier vaccine. The invention also discloses an application of the vaccine on the aspect of treating primary hypertension. Thevaccine can generate one section of efficient specific immunogenic peptide resisting human angiotensin II receptor 1 type and the antibody of the derivant thereof. RAS (Renin-Angiotensin System) can be effectively prevented from activating, the blood pressure of spontaneously hypertensive rats can be obviously lowered, and the vector vaccine has good function on protecting target organs.

Description

field of invention [0001] The invention belongs to the field of biotechnology medicine and biotherapeutics, and in particular relates to an immunogenic peptide segment of angiotensin II receptor type 1 (AT1R) and its derivatives and a carrier, especially a virus-like particle (VLP) The polypeptide-carrier vaccine prepared after coupling, and the effect of the vaccine in treating essential hypertension. Background of the invention [0002] Hypertension is a worldwide problem that seriously endangers human health. The prevalence rate of hypertension in China is 18.8% (2002), and there are nearly 200 million hypertensive patients in the country at present. Hypertension is the main risk factor for cardiovascular and cerebrovascular diseases. The incidence of stroke in China is about 250 / 100,000 people, and the incidence of coronary heart disease is about 50 / 100,000. Actively controlling high blood pressure is an important measure to prevent stroke and coronary heart disease. At...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61K39/00A61P9/12
Inventor 廖玉华周子华陈霄邱志华杨仕俊陈芬王敏余淼
Owner WUHAN HUAJIYUAN BIOTECH DEV
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