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Imatinib immunogen, derivative, synthesis method, specific antibody and detection reagent and preparation methods

A technology of imatinib and its derivatives, which is applied in biological testing, immunoglobulin, biomaterial analysis, etc., can solve the problems that are not suitable for large-scale clinical application and cannot meet the needs of clinical testing, so as to reduce testing costs and combine Powerful, accurate results

Active Publication Date: 2015-07-29
长沙博源医疗科技有限公司
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AI Technical Summary

Problems solved by technology

The detection and analysis of imatinib mainly include HPLC-UV, LC-MS-MS, MESED-LC-MS-MS, C 14 Radioactivity measurement, capillary electrophoresis analysis, HPLC-ESI-MS and electrochemical analysis, etc., but these methods are not suitable for large-scale clinical application, and cannot meet the growing demand for clinical testing
At present, there is a lack of imatinib detection reagents with good stability, high sensitivity and strong specificity in the market, especially good quality automatic detection reagents

Method used

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  • Imatinib immunogen, derivative, synthesis method, specific antibody and detection reagent and preparation methods
  • Imatinib immunogen, derivative, synthesis method, specific antibody and detection reagent and preparation methods
  • Imatinib immunogen, derivative, synthesis method, specific antibody and detection reagent and preparation methods

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Experimental program
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Embodiment 1

[0067] Synthesis and Quantitative Detection of Embodiment 1 Imatinib Derivatives

[0068] The chemical structure of imatinib derivative is shown in formula (IV):

[0069]

[0070] The synthetic route and preparation steps of the above-mentioned imatinib derivatives are as follows:

[0071]

[0072] Concrete synthetic steps are as follows:

[0073] Synthesis of compound 3

[0074]

[0075] Weigh 20g (87.31mmol) of compound 1, dissolve in CH 3 CN, then weigh 18g (130.96mmol) K 2 CO 3 and 19.5g (104.77mmol) of compound 2 (1-tert-butoxycarbonylpiperazine) were sequentially added to the above solution to prepare a reaction mixture solution. After the addition of all ingredients was complete, the above reaction mixture was stirred overnight at 60°C. The solvent was evaporated by a reduced pressure method, and then the obtained residue was dissolved in 300mL of EA, washed with 60mL of brine, and finally the organic phase was passed through Na 2 SO 4 After drying and ...

Embodiment 2

[0095] The synthesis of embodiment two imatinib immunogens

[0096] The imatinib immunogen is composed of bovine serum albumin (Bovine Serum Albumin, BSA) and imatinib derivatives represented by formula (II) -(CH 2 ) n The -COO- group is connected. In this embodiment, the synthesis method of the immunogen is described in detail by taking n=4 as an example. The specific steps are as follows:

[0097] 1. Dissolve 200mg bovine serum albumin in 50ml 0.2M, pH 8.5 phosphate buffer;

[0098] 2. Add the following chemicals into a small beaker and stir to dissolve: 200mg of synthetic imatinib derivatives, 3.5ml of dimethylformamide, 3.5ml of ethanol, 7.0ml of 10mM potassium phosphate buffer at pH 5.0, 200mg of 1 -Ethyl-3-(-3-dimethylaminopropyl)carbodiimide, 50mg N-hydroxysulfosuccinimide, these chemicals were stirred and dissolved at room temperature for 30min;

[0099] 3. The dissolved solution was added dropwise to the BSA solution, and stirred overnight at 2-8°C to obtain the an...

Embodiment 3

[0100] Example 3: Preparation of Anti-Imatinib Specific Antibody

[0101] The imatinib 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:

[0102] 1. Dilute the above synthesized imatinib 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.

[0103] 2. After 2-3 weeks, inject 1.0ml of the same antigen solution and incomplete Freund's adjuvant to the above-mentioned experimental animal rabbit once, and then inject once every four weeks, a total of 4 injections.

[0104] 3. Take blood from the experimental animal rabbit in step 2, separate and purify to obtain anti-imatinib specific antibody with a titer of 1:30000-1:50000.

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Abstract

The invention discloses an imatinib immunogen, derivative, synthesis method, specific antibody and detection reagent and preparation methods. The imatinib immunogen prepared by the invention is high in immunogenicity, can be induced to obtain a high-titer anti-imatinib specific antibody and does not crossly react with sixty-two common drugs; the imatinib detection reagent prepared by using the antibody can accurately and quickly determine the content of imatinib in samples. Compared with the existing detection reagent in the market, the detection reagent disclosed by the invention has the advantages of simplicity and convenience in operation, high sensitivity, high specificity, accurate result and the like, the imatinib detection cost can be effectively reduced and the large-scale clinical popularization and use are facilitated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to imatinib immunogens, derivatives and synthesis methods, specific antibodies, detection reagents and preparation methods. Background technique [0002] Imatinib (Imitimib), its structural formula is shown in formula (Ⅲ): [0003] [0004] Imatinib is an anticancer drug, and its pharmacologically active salt: imatinib mesylate (trade name Gleevec), is mainly used for the treatment of chronic myelogenous leukemia, especially for the treatment failure of α-interferon Patients with chronic myelogenous leukemia in blast crisis, accelerated or chronic stages. In addition, this product can also be used for the treatment of various diseases such as gastrointestinal stromal cell tumor, dermatofibrosarcoma protuberans, Philadelphia chromosome-positive acute lymphoblastic leukemia, eosinophilia, and systemic mastocytosis. The pharmacokinetics of imatinib and its salts have been reported to be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K14/795C07K14/435C07K16/44C07D401/04G01N33/53G01N33/535
CPCC07D401/04C07K14/435C07K14/765C07K14/795C07K16/44C07K19/00G01N33/535G01N33/94G01N2500/04
Inventor 虞留明李振荣曹永彤王海滨
Owner 长沙博源医疗科技有限公司
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