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Topiramate derivative, immunogen, anti-topiramate specific antibody and preparation method and application thereof

A technology for topiramate and derivatives, applied in the field of topiramate derivatives, can solve the problems of difficult control of conditions, low quantitative lower limit, wide linear range and the like, and achieve the effects of accurate results, high sensitivity and improved immunogenicity

Pending Publication Date: 2022-07-01
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 can greatly simplify the sample purification process, and the results are not interfered by topiramate metabolites, and have a wide linear range and a low lower limit of quantitation, but the requirements for the test are high, the pretreatment is more complicated, and the conditions are not easy to control

Method used

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  • Topiramate derivative, immunogen, anti-topiramate specific antibody and preparation method and application thereof
  • Topiramate derivative, immunogen, anti-topiramate specific antibody and preparation method and application thereof
  • Topiramate derivative, immunogen, anti-topiramate specific antibody and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1: Synthesis of Topiramate Derivatives

[0082] Topiramate derivatives were synthesized by the following synthetic routes:

[0083]

[0084] The specific synthesis steps are as follows:

[0085] (1) Synthesis of compound 2:

[0086]

[0087] Compound 1 (10 g, 38.46 mmol) and Et3N (5.8 g, 57.69 mmol) were dissolved in DCM (100 mL), followed by the addition of sulfamoyl chloride (6.6 g, 57.69 mmol) to make a reaction mixture, which was prepared in Stir overnight at room temperature. After the reaction, the reaction mixture was washed with purified water, brine, and then passed through Na 2 SO 4 Dry and concentrate. The residue after solvent removal was purified by silica gel chromatography to obtain 5.0 g of compound 2 as a white solid.

[0088] (2) Synthesis of Levetiracetam Derivatives:

[0089]

[0090] Compound 2 (5.0 g, 14.75 mmol) was dissolved in THF (50 mL), then dihydropyran-2,6-dione (5.0 g, 44.25 mmol) and potassium tert-butoxide (5.0 g,...

Embodiment 2

[0091] Example 2: Preparation of Topiramate Immunogen

[0092] The specific steps of the preparation method of topiramate immunogen are as follows:

[0093] (1) Preparation of carrier protein solution: Dissolve recombinant camel serum albumin in 0.35mol / L potassium phosphate buffer (pH=8.5), and the final concentration of recombinant camel serum albumin is 5.0mg / mL to obtain carrier protein solution;

[0094] (2) Preparation of topiramate derivative solution: 250.0 mg of the above-mentioned topiramate derivative, 7.5 mL of dimethylformamide, 7.5 mL of ethanol, 15.0 mL of potassium phosphate buffer (10.0 mmol / L, pH=8.0), 150.0 mL of mg 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide and 90.0 mg N-hydroxysulfosuccinimide were mixed, and stirred and dissolved for 3 hours to obtain a topiramate derivative solution;

[0095] (3) Synthesis of topiramate immunogen: add the topiramate derivative solution obtained in step (2) dropwise to the carrier protein solution obtained in step ...

Embodiment 3

[0096] Example 3: Preparation of Anti-Topiramate Specific Antibodies

[0097] The specific steps of the preparation method of anti-topiramate-specific antibody are as follows:

[0098] (1) Dilute the above-mentioned topiramate 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. The complete adjuvant was mixed, and the experimental animals were injected into rabbits at multiple points;

[0099] (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;

[0100] (3) Blood is collected from the experimental animal rabbits injected in step (2), separated and purified to obtain an anti-topiramate specific antib...

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Abstract

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

Description

technical field [0001] The invention relates to a topiramate derivative, an immunogen, an anti-topiramate specific antibody, a preparation method and application thereof, and belongs to the technical field of biomedical detection. Background technique [0002] Topiramate (TPM) is a new type of antiepileptic drug with a chemical structure of 2,3:4,5-bis-0(1-methylethylene)-β-dextro-sulfamate fructopyranose . Topiramate has a completely different structural formula from other antiepileptic drugs. It is the thiol of natural monosaccharide-based dextrose, its molecular formula is: C12H12NO8S, and its relative molecular weight is: 339.36. Topiramate is mainly used for the treatment of simple and complex partial seizures and generalized tonic-clonic seizures. Clinically, monotherapy or as an additive drug on the basis of the original antiepileptic drug have shown good effects. It has been widely used in Europe and the United States to treat various types of epilepsy, and now it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D493/14C07K14/765C07K16/44C07K16/06G01N33/546G01N33/543G01N33/535
CPCC07D493/14C07K14/765C07K16/44C07K16/065G01N33/94G01N33/54313G01N33/535
Inventor 张小可王翰博
Owner 长沙博源医疗科技有限公司
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