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Oxcarbazepine derivative, immunogen, anti-oxcarbazepine specific antibody as well as preparation method and application of oxcarbazepine specific antibody

A technology of oxcarbazepine and its derivatives, which is applied in the preparation method of peptides, biological testing, immunoglobulin, etc., can solve the problems of cumbersome operation, inconvenient large-scale clinical testing, and difficult control of the process, and achieve high sensitivity and effective Conducive to clinical promotion and use, high coupling efficiency

Pending Publication Date: 2022-06-28
长沙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

Liquid chromatography-tandem mass spectrometry simultaneously determines the concentration of oxcarbazepine and MHD in human plasma, the amount of sample pretreatment plasma is small, and the analysis time is short, but the operation is cumbersome, the process is not easy to control, and it is inconvenient to form a large-scale clinical test

Method used

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  • Oxcarbazepine derivative, immunogen, anti-oxcarbazepine specific antibody as well as preparation method and application of oxcarbazepine specific antibody
  • Oxcarbazepine derivative, immunogen, anti-oxcarbazepine specific antibody as well as preparation method and application of oxcarbazepine specific antibody
  • Oxcarbazepine derivative, immunogen, anti-oxcarbazepine specific antibody as well as preparation method and application of oxcarbazepine specific antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1: Synthesis of Oxcarbazepine Derivatives

[0097] Oxcarbazepine derivatives were synthesized by the following synthetic routes:

[0098]

[0099] The specific synthesis steps are as follows:

[0100] (1) Synthesis of compound 2:

[0101]

[0102] Weigh 2.5 g of potassium nitrosodisulfonate (Fremy's salt, (KSO 3 ) 2 NO), 9.32 mmol) and 1.8 g Na 2 HPO 4 (12.7 mmol), put it into a beaker, add 95 ml of double distilled water to dissolve, adjust the pH to 7.22; weigh 0.6 g of compound 1 (12.76 mmol) into 60 ml of acetone solution; mix the above two solutions and stir vigorously to obtain a purple color solution; the above purple solution was added to the acetone solution, stirred continuously for 10 minutes, filtered, and placed in a refrigerator overnight; the overnight solution was concentrated by an argon rotary evaporator, extracted with 500 ml of ether, and after the extraction was completed, the organic The phase solvent was evaporated; the evapora...

Embodiment 2

[0121] Example 2: Preparation of Oxcarbazepine Immunogen

[0122] The specific steps of the preparation method of oxcarbazepine immunogen are as follows:

[0123] (1) Preparation of carrier protein solution: Dissolve recombinant human serum albumin in 0.35mol / L potassium phosphate buffer (pH=8.5), and the final concentration of recombinant human serum albumin is 5.0mg / mL to obtain carrier protein solution;

[0124] (2) Preparation of oxcarbazepine derivative solution: 250.0 mg of the above oxcarbazepine derivative, 7.5 mL of dimethylformamide, 7.5 mL of ethanol, and 15.0 mL of potassium phosphate buffer (10.0 mmol / L, pH= 8.0), 150.0 mg of 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide, and 90.0 mg of N-hydroxysulfosuccinimide were mixed, stirred and dissolved for 3 hours to obtain oxcarbazepine Derivative solution;

[0125] (3) Synthesis of oxcarbazepine immunogen: The oxcarbazepine derivative solution obtained in step (2) was added dropwise to the carrier protein solution...

Embodiment 3

[0126] Example 3: Preparation of anti-oxcarbazepine-specific antibodies

[0127] The specific steps of the preparation method of anti-oxcarbazepine-specific antibody are as follows:

[0128] (1) Dilute the above oxcarbazepine immunogen with 0.15mol / L sodium phosphate buffer (pH=7.0) to a final concentration of 3.5mg / mL to obtain an artificial antigen solution, and then mix 3.0mL of the artificial antigen solution with an equal amount of Freund's complete adjuvant was mixed and injected into experimental animals rabbits at multiple points;

[0129] (2) After 4 weeks, 3.0 mL of the same artificial antigen solution and the same amount of incomplete Freund's adjuvant were injected into the above-mentioned experimental animal rabbits at multiple points, and then once every 5 weeks, for a total of 6 injections;

[0130] (3) Blood is collected from the experimental animal rabbit that has been injected in step (2), and is separated and purified to obtain an anti-oxcarbazepine-specifi...

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Abstract

The invention discloses an oxcarbazepine derivative, an immunogen, an anti-oxcarbazepine specific antibody as well as a preparation method and application of the oxcarbazepine specific antibody. Firstly, a novel oxcarbazepine derivative and recombinant human serum albumin obtained through genetic engineering modification are coupled to prepare an oxcarbazepine artificial antigen, then the oxcarbazepine artificial antigen is used for immunizing an experimental animal to obtain an anti-oxcarbazepine specific antibody, and ELISA detection shows that the specific antibody is high in specificity and sensitivity and can be used for preparing the oxcarbazepine specific antibody. An interference experiment shows that the specific antibody does not have any cross reaction with 100 common drugs; the anti-oxcarbazepine specific antibody is applied to preparation of an oxcarbazepine detection reagent, the oxcarbazepine detection reagent comprises an oxcarbazepine average phase enzyme immunoassay reagent and an oxcarbazepine latex enhanced immunoturbidimetry detection reagent, and the detection reagent can realize high-flux and rapid detection of oxcarbazepine on a full-automatic biochemical analyzer.

Description

technical field [0001] The invention relates to an oxcarbazepine derivative, an immunogen, an anti-oxcarbazepine specific antibody, a preparation method and application thereof, and belongs to the technical field of biomedical detection. Background technique [0002] Oxcarbazepine is an antiepileptic drug approved as an adjunctive or monotherapy for the treatment of partial-onset epilepsy in adults and children. The chemical structure of oxcarbazepine is 10,11-dihydro-10-oxo-5H-dibenzo[b,f]azepine-5-carboxamide, and the chemical structure of its metabolite 10-hydroxycarbamazepine It is 10,11-dihydro-10-hydrocarbamazepine. Oxcarbazepine is soluble in DMSO, methanol, water, ethanol and acetone, and the solubility in DMSO is ≤9 mg / mL. The solubility of 10-hydroxycarbamazepine in methanol is 100 μg / mL. Oxcarbazepine is rapidly absorbed through the gastrointestinal tract and rapidly degraded to a pharmacologically active metabolite, 10-hydroxycarbamazepine (MHD). Several stud...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D223/22C07K14/765C07K1/02C07K1/107C07K1/34C07K16/44C07K16/06G01N33/68G01N33/535G01N33/532
CPCC07D223/22C07K14/765C07K16/44C07K16/065G01N33/94G01N33/6854G01N33/535G01N33/532
Inventor 张小可张韶斌陈斯亮
Owner 长沙博源医疗科技有限公司
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