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Dipeptide derivatives for detection of angiotensin I, preparation method and application thereof

An angiotensin and anti-angiotensin technology, applied in the fields of peptides, measuring devices, instruments, etc., can solve the problems of low detection sensitivity, poor specificity, and inability to detect, and achieve high immunogenicity, high accuracy, and full realization. automated effects

Active Publication Date: 2020-08-07
苏州博源医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is: the angiotensin Ⅰ detection reagent in the prior art has low detection sensitivity and poor specificity, and there is no leucine-histidine dipeptide derivative for detecting angiotensin Ⅰ, which cannot It is detected by homogeneous enzyme reagents, with low detection sensitivity and poor specificity

Method used

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  • Dipeptide derivatives for detection of angiotensin I, preparation method and application thereof
  • Dipeptide derivatives for detection of angiotensin I, preparation method and application thereof
  • Dipeptide derivatives for detection of angiotensin I, preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Embodiment 1: the synthesis of leucine-histidine dipeptide derivative

[0056] The chemical structure of the leucine-histidine dipeptide derivative is shown in formula (I):

[0057]

[0058] The synthetic route and preparation steps of the above-mentioned leucine-histidine dipeptide derivatives are as follows:

[0059]

[0060] Concrete synthetic steps are as follows:

[0061] (1) Synthesis of compound 2

[0062] Weigh 25g (86.2mmol) of compound 1 and dissolve it in 250mL of dichloromethane (DCM), then add 17.2g (103.5mmol) of compound A, 26.5g (172.4mmol) of 1-hydroxybenzotriazole (HOBT ), 38g (172.4mmol) of carbodiimide (EDCI), and 50g (500mmol) of triethanolamine (TEA) were prepared as reaction mixture solution I. The reaction mixture I was stirred overnight at 30°C. After the reaction, an appropriate amount of purified water was added to the reaction mixture solution I, and then 150 mL of DCM was used for extraction, and the extraction step was repeated 4 ...

Embodiment 2

[0074] Example 2: Synthesis of Leucine-Histidine Dipeptide Immunogen

[0075] The leucine-histidine dipeptide immunogen is composed of bovine serum albumin (Bovine Serum Albumin, BSA) and -CO-(CH 2 ) 3 The -CO- group is connected, and the structural formula is shown in the following formula (II):

[0076]

[0077] The concrete steps of the synthetic method of this immunogen are as follows:

[0078] 1. Dissolve 200mg bovine serum albumin in 50ml 0.2M, pH 8.5 phosphate buffer;

[0079] 2. Add the following chemicals into a small beaker and stir to dissolve: 200mg of the leucine-histidine dipeptide derivative synthesized in Example 1, 3.5ml of dimethylformamide, 3.5ml of ethanol, 7.0ml of 10mM, pH 5.0 Potassium phosphate buffer solution, 200mg 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide, 50mg N-hydroxyl sulfosuccinimide, these chemicals were stirred and dissolved at room temperature for 30min ;

[0080] 3. Add the dissolved solution dropwise to the BSA solution, and s...

Embodiment 3

[0082] Example 3: Preparation of Anti-Angiotensin I Specific Antibody

[0083] The leucine-histidine dipeptide immunogen prepared in Example 2 was inoculated into experimental animal rabbits by conventional methods, and the antiserum was taken after booster immunization. The specific steps were as follows:

[0084] 1. Dilute the above synthesized leucine-histidine dipeptide immunogen to 1.0 mg / ml with PBS to obtain an antigen solution, then mix 1.0 ml of the antigen solution with Freund's complete adjuvant, and inject the experimental animal rabbit .

[0085] 2. After 2 to 3 weeks, inject 1.0ml of the same antigen solution and incomplete Freund's adjuvant to the above-mentioned experimental rabbits once, and then inject once every four weeks, a total of 4 injections.

[0086] 3. Take blood from the experimental animal rabbit in step 2, separate and purify to obtain anti-angiotensin I specific antibody with a titer of 1:30000-1:50000.

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Abstract

The invention relates to a dipeptide derivative for detecting angiotensin I as well as a preparation method and application thereof. The dipeptide derivative is a leucine-histidine dipeptide derivative and a structural formula is shown as a formula (I) as follows: the formula (I) is shown in the description. According to the dipeptide derivative provided by the invention, the leucine-histidine dipeptide derivative, repeated tests are carried out and an immunogen and a corresponding antibody are prepared in a pioneering manner by selecting two amino acid residues at the tail end of carboxyl ofthe angiotensin I; the obtained antibody can be used for identifying and combining the angiotensin I and a plurality of samples can be simultaneously determined on a full-automatic biochemical analyzer, so that high-throughput and rapid determination of the angiotensin I is realized; the dipeptide derivative has high accuracy and strong specificity; the precision and the detection efficiency are greatly improved when being compared with previous precision and detection efficiency; meanwhile, full automation of a detection process is realized, requirements on inspectors are not high and the dipeptide derivative is easy to realize, popularize and apply.

Description

technical field [0001] The invention relates to a dipeptide derivative used for detecting angiotensin I, a preparation method and application thereof, and belongs to the technical field of biological detection. Background technique [0002] The chemical structural formula of angiotensin I is shown in formula (Ⅲ): [0003] [0004] Hypertension is a worldwide common disease with a high incidence and serious harm to human health. The prevalence of the global population is as high as 10% to 20%. Hypertension is an independent risk factor for cardiovascular and cerebrovascular diseases, which can lead to lesions of cardiovascular and cerebrovascular, heart, kidney, etc., and is one of the main diseases that endanger human health. According to the latest data from the National Health and Family Planning Commission, there are more than 200 million hypertensive patients in my country. With the development of the economy and the improvement of people's living standards, hyperten...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/078
CPCG01N33/535G01N33/543
Inventor 余琳程黎明虞留明
Owner 苏州博源医疗科技有限公司
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