Drug target expression and purification method for high blood pressure

A purification method and high blood pressure technology, which is applied in botany equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve the problems of large side effects and lack of specificity of antihypertensive drugs, and achieve the promotion of cell growth , reduce cell aggregation, and promote the effect of research and development

Inactive Publication Date: 2020-11-20
江西晶美瑞生物医药有限公司
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Problems solved by technology

[0005] In order to solve the defects of lack of specificity and large side effects of antihypertensive drugs on the market, we designed corresponding specific small molecule inhibitors and related antibodies for TRPM4, a key drug target of hypertension, to treat hypertension, etc. The treatment of cardiovascular related diseases has important theoretical significance and practical application value

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  • Drug target expression and purification method for high blood pressure

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Embodiment Construction

[0025] In order to make the object, technical solution and technical effect of the present invention clearer, the present invention will be further described below in conjunction with specific embodiments. It should be understood that the specific embodiments and related drawings described here are only used to explain the present invention, not to limit the present invention.

[0026] 1. Expression of drug targets

[0027] ①Construction of drug target expression plasmid

[0028] The human full-length TRPM4 construct, whose amino acid sequence is shown in Sequence No.1, was cloned into the pAntigenMem vector, and the pAntigenMem vector map is shown in figure 1 shown. A maltose-binding protein (MBP) tag (to improve the solubility of the drug target) was added to its N-terminus. The constructed recombinant plasmid was transformed into DH5α competent cells, plated on LB plates containing a certain concentration of antibiotics, colonies were selected, plasmids were extracted, a...

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Abstract

The invention relates to the technical field of biology, in particular to a drug target expression and purification method for high blood pressure. The drug target expression and purification method for high blood pressure comprises following steps: a human full-length TRPM4 gene segment is cloned to a pAntigen Mem vector, positive clones are screened; plasmids are extracted, competent DH10 strains are transformed, and positive clones are selected; rod granules are extracted, and P3 baculoviruses are produced in a baculovirus expression system; the P3 baculoviruses are transduced into HEK293SGnTI<-> cells, induced expression is performed, and purified drug targets are collected. After that, according to the TRPM4 drug target for the high blood pressure, design of corresponding specific small-molecule inhibitors and related antibodies is facilitated, and the method is applied to treatment of cardiovascular diseases such as the high blood pressure and the like. The drug target expression and purification method for the high blood pressure can produce the drug targets with normal biological activity and yield up to milligram level for the high blood pressure, and promote corresponding research and development of related antibodies.

Description

technical field [0001] The invention relates to the technical field of hypertension drugs, in particular to a method for expressing and purifying drug targets for hypertension. Background technique [0002] Cardiovascular disease (CVD), such as hypertension, stroke and coronary artery disease, is one of the most serious diseases threatening human beings in the world today, and has become an increasingly urgent major public health problem. Among them, hypertension is one of the most common cardiovascular diseases, and its incidence is increasing year by year, and the clinical application of antihypertensive drugs is also constantly developing and updating. [0003] Drugs clinically used in the treatment of hypertension mainly include calcium antagonists (CCB), angiotensin II receptor blockers (ARB), β-blockers, angiotensin converting enzyme inhibitors (ACEI) and diuretics. These medicines all have different degrees of toxic and side effects, and there are great application l...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/866C12N15/12C12P21/02
CPCC07K14/47C12N15/86C12N2710/14043C12N2800/107
Inventor 张进李健万双燕
Owner 江西晶美瑞生物医药有限公司
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