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Olanzapine derivative, immunogen, anti-olanzapine specific antibody and preparation method and application thereof

A technology for olanzapine and its derivatives, which is applied in the field of olanzapine derivatives, can solve problems such as difficult control of conditions, high requirements and costs, complex pretreatment, etc., achieve accurate results, reduce detection costs, and improve immunogenicity Effect

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 olanzapine metabolites, and have a wide linear range and a low lower limit of quantitation, but the requirements for the test and the cost are high, and the pre-treatment is relatively complicated , the conditions are not easy to control

Method used

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  • Olanzapine derivative, immunogen, anti-olanzapine specific antibody and preparation method and application thereof
  • Olanzapine derivative, immunogen, anti-olanzapine specific antibody and preparation method and application thereof
  • Olanzapine derivative, immunogen, anti-olanzapine specific antibody and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1: Synthesis of Olanzapine Derivatives

[0084] Olanzapine derivatives were synthesized by the following synthetic routes:

[0085] ;

[0086] The specific synthesis steps are as follows:

[0087] (1) Synthesis of compound 2:

[0088]

[0089] Compound 1 (6 g, 26 mmol) and piperazine (18 g, 0.19 mol) were added to butanol (100 mL) at room temperature to prepare a reaction mixture, and the reaction mixture was stirred under heating at 125°C for 18 hours. After the reaction was complete, the above reaction mixture was concentrated in vacuo. The concentrated residue was purified by flash chromatography (methanol:dichloromethane 0-10%) to give 6 g of compound 2 as a yellow solid.

[0090] (2) Synthesis of compound 3:

[0091]

[0092] Compound 2 (6 g, 20 mmol), tert-butyl 5-bromovalerate (5.7 g, 24 mmol), K 2 CO 3 (4.2 g, 28 mmol), KI (0.3 g, 2 mmol) were added to acetonitrile (100 mL) at room temperature to make a reaction mixture. The reaction mixt...

Embodiment 2

[0096] Example 2: Preparation of Olanzapine Immunogen

[0097] The specific steps of the preparation method of olanzapine immunogen are as follows:

[0098] (1) Preparation of carrier protein solution: Dissolve recombinant sheep serum albumin in 0.35 mol / L potassium phosphate buffer (pH=8.5), and the final concentration of recombinant sheep serum albumin is 5.0 mg / mL to obtain carrier protein solution;

[0099] (2) Preparation of olanzapine derivative solution: 250.0 mg of the above-mentioned olanzapine 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 olanzapine Derivative solution;

[0100] (3) Synthesis of olanzapine immunogen: The olanzapine derivative solution obtained in step (2) was added dropwise to the carrier protein solution obtained...

Embodiment 3

[0101] Example 3: Preparation of anti-olanzapine-specific antibodies

[0102] The specific steps of the preparation method of anti-olanzapine-specific antibody are as follows:

[0103] (1) Dilute the above-mentioned olanzapine 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;

[0104] (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;

[0105] (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-olanzapine-specific ...

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Abstract

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

Description

technical field [0001] The invention relates to an olanzapine derivative, an immunogen, an anti-olanzapine specific antibody, a preparation method and application thereof, and belongs to the technical field of biomedical detection. Background technique [0002] Olanzapine is a novel atypical neuroleptic that binds to dopamine receptors, serotonin (5-HT) receptors and cholinergic receptors and exerts an antagonistic effect. Olanzapine is mainly used clinically for the acute phase and maintenance treatment of schizophrenia and other psychoses with severe positive symptoms / or negative symptoms; it can also be used in relieving secondary affective symptoms common to schizophrenia and related diseases. play a certain role. The chemical name for olanzapine is 2-methyl-4-(4-methyl-1-piperazinyl)-10H-thieno[2,3-b][1,5]benzodiazepine Zhuo, closed the ring with sulfur, propionaldehyde and malononitrile as raw materials, and then reacted with o-nitrofluorobenzene to obtain 2-(2-nitro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D495/04C07K14/765C07K1/02C07K1/107C07K1/34C07K16/06C07K16/44G01N33/94G01N33/68G01N33/535G01N33/543
CPCC07D495/04C07K14/765C07K16/44C07K16/065G01N33/94G01N33/6854G01N33/535G01N33/54313
Inventor 张小可王永霞封飒飒胡振阳
Owner 长沙博源医疗科技有限公司
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