Dipeptide derivative for detection of angiotensin Ⅱ and preparation method and application thereof

A derivative, phenylalanine dipeptide technology, applied in biological testing, biomaterial analysis, peptides, etc., can solve the problems of low detection sensitivity, poor specificity, and undetectability, and achieve high immunogenicity and high accuracy , The effect of improving the detection efficiency

Active Publication Date: 2020-06-30
苏州博源医疗科技有限公司
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  • 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 II detection reagent in the prior art has low detection sensitivity and poor specificity, and there is no proline-phenylalanine dipeptide derivative for detecting angiotensin II, It cannot be detected by homogeneous enzyme reagents, with low detection sensitivity and poor specificity

Method used

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  • Dipeptide derivative for detection of angiotensin Ⅱ and preparation method and application thereof
  • Dipeptide derivative for detection of angiotensin Ⅱ and preparation method and application thereof
  • Dipeptide derivative for detection of angiotensin Ⅱ and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. Synthesis of proline-phenylalanine dipeptide derivatives

[0057] The chemical structure of proline-phenylalanine dipeptide derivative is shown in formula (I):

[0058]

[0059] The synthetic route and preparation steps of the above-mentioned proline-phenylalanine dipeptide derivatives are as follows:

[0060]

[0061] Concrete synthetic steps are as follows:

[0062] (1) Synthesis of Compound 2

[0063] Weigh 40g (93.6mmol) of compound 1 and dissolve it in 400mL of DCM, then add 24g (112mmol) of compound A, 25.2g (187.2mmol) of 1-hydroxybenzotriazole (HOBT), 36g (187.2mmol) Carbodiimide (EDCI) and 47.2g (480mmol) 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 times. The extracted orga...

Embodiment 2

[0075] Example 2. Synthesis of Proline-Phenylalanine Dipeptide Immunogen

[0076] The proline-phenylalanine dipeptide immunogen is composed of bovine serum albumin (Bovine Serum Albumin, BSA) and -CO-(CH 2 ) 3 It is formed by connecting -CO- groups, and its structural formula is shown in the following formula (II):

[0077]

[0078] The concrete steps of the synthetic method of this immunogen are as follows:

[0079] 1. Dissolve 200mg bovine serum albumin in 50ml 0.2M, pH 8.5 phosphate buffer;

[0080] 2. Add the following chemicals into a small beaker and stir to dissolve: 200mg of the proline-phenylalanine dipeptide derivative synthesized in Example 1, 3.5ml of dimethylformamide, 3.5ml of ethanol, 7.0ml of 10mM, pH 5.0 potassium phosphate buffer, 200mg 1-ethyl-3-(-3-dimethylaminopropyl)carbodiimide, 50mg N-hydroxysulfosuccinimide, these chemicals were stirred and dissolved at room temperature React for 30 minutes;

[0081] 3. Add the dissolved solution dropwise to th...

Embodiment 3

[0082] Example 3. Preparation of anti-angiotensin II specific antibody

[0083] The proline-phenylalanine 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 proline-phenylalanine dipeptide immunogen synthesized above 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 conduct the experiment on rabbits injection.

[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 II specific antibody with a titer of 1:30000-1:50000.

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Abstract

The invention relates to a dipeptide derivative for detecting Ang II and a preparation method and application of the dipeptide derivative for detecting Ang II. The dipeptide derivative is a proline-phenylalanine dipeptide derivative, and has the structural formula as shown in the following formula (I). The proline-phenylalanine dipeptide derivative is prepared pioneeringly by selecting two amino acid residues at the carboxyl terminal of Ang II through repeated experiments, highly immunogenic small-molecule immunogen and a corresponding antibody are obtained through the prepared proline-phenylalanine dipeptide derivative, a homogeneous enzyme immunoassay reagent containing the anti-Ang II specific antibody can conveniently, quickly, and accurately determine the content of Ang II in a biological sample, and multiple samples can be determined at the same time on an automatic biochemical analyzer to realize high-throughput and rapid determination of Ang II, so that the precision is high, the specificity is strong and the accuracy and detection efficiency are greatly improved compared with the previous accuracy and detection efficiency.

Description

technical field [0001] The invention relates to a dipeptide derivative used for detecting angiotensin II and its preparation method and application, belonging to the technical field of biological detection. Background technique [0002] The chemical structural formula of angiotensin II 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 rate 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 cardiovascular and cerebrovascular diseases. Diseases of heart, kidney, etc. are 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, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/078G01N33/68
CPCG01N21/31G01N33/531G01N33/535G01N33/5436G01N33/6854G01N33/6893G01N2800/321
Inventor 陆丽华陈渡波虞留明
Owner 苏州博源医疗科技有限公司
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