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Vascular smooth muscle L type calcium ion channel immunogenic peptide and application thereof

A calcium ion channel, vascular smooth muscle technology, applied in the preparation of vascular smooth muscle L-type calcium ion channel immunogenic peptide segment, the application field of treating essential hypertension, can solve the problem of lowering blood pressure, low control rate of hypertension treatment, Patients with poor treatment compliance and other problems to achieve the effect of lowering blood pressure

Inactive Publication Date: 2012-07-18
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because chemical drugs need to be taken regularly every day, the patient's treatment compliance is not good, resulting in a low rate of hypertension treatment control. The development of immunology provides a new way for the treatment of hypertension. So far, the vaccines used for the treatment of hypertension mainly target In the RAS system, the CYT006-AngQβ vaccine from Cytos, Switzerland has completed Phase IIa clinical trials and has a good antihypertensive effect
At present, there is no relevant report on the use of active or passive immunization methods to block calcium ion channels in order to reduce blood pressure.

Method used

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  • Vascular smooth muscle L type calcium ion channel immunogenic peptide and application thereof
  • Vascular smooth muscle L type calcium ion channel immunogenic peptide and application thereof
  • Vascular smooth muscle L type calcium ion channel immunogenic peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0017] Implementation Case 1 Preparation and Identification of QβCa-001 Vaccine

[0018] The short peptide CPAEEDPS is directly synthesized by Shanghai Gil Biochemical Co., Ltd., and the purity of the short peptide is 95%.

[0019] The vector Qβ-2aa virus-like particle is constructed, expressed and purified by our laboratory, with a purity of over 90% (a national invention patent has been applied for, patent application number: 201010028904.7). It is prepared as follows:

[0020] 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;

[0021] 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 st...

Embodiment example 2

[0029] Implementation Case 2 Specificity of Anti-QβCa-001 Vaccine Short Peptide Antibody

[0030] Anti-peptide antibody Anti-CN-8 of QβCa-001 vaccine was purified, and the specificity of Anti-CN-8 antibody was identified by immunoblotting and cellular immunofluorescence. Anti-CN-8 is the anti-peptide antibody of animals immunized with QβCa-001 vaccine, and Con is the negative control antibody. The total protein of SD rat mesenteric tertiary arterial smooth muscle cells was extracted and analyzed by Western blot. The results showed that Anti-CN-8 could bind to a protein of about 240kDa in the total protein extract, which was associated with rat L-type calcium channel α1c subunit molecular weight (about 240kDa) consistent ( figure 2 A). At the same time, it was found by cell immunofluorescence that Anti-CN-8 could bind to the L-type calcium channel on the surface of rat mesenteric tertiary artery smooth muscle cells, and the control antibody had basically no binding to the L-...

Embodiment example 3

[0031] Example 3 Effect of Anti-CN-8 Antibody on the Stimulation of Calcium Channel Opener Bay K8644

[0032] The effect of Anti-CN-8 on the increase of calcium ion concentration in vascular smooth muscle cells stimulated by the opener Bay K8644 was observed by laser confocal microscope. f 0 , F maxRepresent the basal and maximum fluorescence intensity of VSMCs after equilibration, respectively. Vascular smooth muscle cells were pre-incubated with Anti-CN-8 and nifedipine for 30 minutes, and then stimulated with Bay K8644. The results showed that Anti-CN-8 could significantly inhibit the increase of calcium ion concentration in vascular smooth muscle cells stimulated by Bay K8644, and the inhibitory effect of nifedipine was more significant (* represents Anti-CN-8 group and Con-Ab group Compared, P image 3 ).

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Abstract

The invention discloses a vascular smooth muscle L type calcium ion channel immunogenic peptide and application thereof, and belongs to the field of biotechnology and medicine. The immunogenic peptide has the following amino acid sequence: CGGPAEEDPS, CPAEEDPS, PAEEDPSGGC or PAEEDPSC, preferably CPAEEDPS. The vascular smooth muscle L type calcium ion channel immunogenic peptide containing a first binding site is coupled and combined with one or more carriers, preferably recombinant Qbeta-2aa phage virus-like particle protein to form ordered and repeated polypeptide-carrier vaccine. The invention discloses application of the vaccine to treatment of primary hypertension. The vaccine can produce high-efficiency specific antibody against the vascular smooth muscle L type calcium ion channel immunogenic peptide. The antibody can be used for effectively obstructing an L type calcium ion channel, so that the blood pressure of a spontaneously hypertensive rat can be obviously reduced.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and in particular relates to the preparation of an immunogenic peptide segment of L-type calcium ion channel of vascular smooth muscle and the application of a vaccine prepared by using the peptide segment in treating essential hypertension. Background technique [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 present, the treatment control rate of outpatient hypertensive patients in China ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K39/385A61K47/48A61P9/12
Inventor 廖玉华陈霄周子华邱志华陈芬杨仕俊林吉斌丁丹王敏
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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