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Oxazepam hapten, oxazepam antigen and preparation method and application thereof

A hapten and antigen technology, applied in the biological field, can solve the problems of being expensive, unable to meet the requirements of fast, convenient, and accurate, and achieve the effects of improving the detection rate, reducing cross-substance interference, and high reaction yield

Inactive Publication Date: 2021-01-05
GUANGZHOU WONDFO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, these methods not only require expensive instruments and equipment, but also have relatively high requirements for inspection materials, which require further purification and treatment, which can no longer meet the requirements of modern detection for speed, convenience, and accuracy.

Method used

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  • Oxazepam hapten, oxazepam antigen and preparation method and application thereof
  • Oxazepam hapten, oxazepam antigen and preparation method and application thereof
  • Oxazepam hapten, oxazepam antigen and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0054] This embodiment prepares a kind of oxazepam hapten, and the preparation method is as follows:

[0055] Weigh 10mmol of anhydrous aluminum trichloride and 5mmol of succinic anhydride in a 50mL round-bottomed flask, add 10mL of dichloromethane, and stir at 0°C; weigh 5mmol of oxazepam, and add in three times to the above reaction After the addition, it was raised to 25°C and reacted for more than 12 hours; under vigorous stirring, slowly added 20mL of ultrapure water, and after standing for stratification, the organic layer was taken for vacuum distillation, and the obtained residue was dissolved in 5ml of ethyl acetate. Wash twice with saturated sodium chloride solution, 5 mL each time, dry the organic layer, evaporate to dryness, and obtain 1.8 mmol hapten after purification by column chromatography. Carry out ESI-MS analysis (385.1[M-1]) to the prepared hapten, the result is as follows figure 1 As shown, it was proved that the oxazepam hapten was successfully prepared...

Embodiment 2

[0058] This embodiment prepares a kind of oxazepam hapten, and the preparation method is as follows:

[0059] Weigh 20mmol of anhydrous aluminum trichloride and 5mmol of succinic anhydride into a 50mL round-bottomed flask, add 10mL of dichloromethane, and stir at 0°C; weigh 5mmol of oxazepam, and add it to the above reaction in 3 times After the addition, it was raised to 25°C and reacted for more than 12 hours; under vigorous stirring, slowly added 20mL of ultrapure water, and after standing for stratification, there were a large amount of flocculent precipitates, and the organic layer was taken for vacuum distillation, and the obtained residual Dissolve in 5ml ethyl acetate, wash twice with saturated sodium chloride solution, 5mL each time, dry the organic layer, evaporate to dryness, and obtain 0.03mmol hapten after purification by column chromatography. The obtained hapten was compared with the hapten obtained in Example 1, and it was found to be the same substance, but th...

Embodiment 3

[0062] This embodiment prepares a kind of oxazepam antigen, and the preparation method is as follows:

[0063] (1) Preparation of oxazepam hapten active ester:

[0064] In a 25 mL dry single-necked bottle, add 1 mmol of the hapten prepared in Example 1, 1.2 mmol of N-hydroxysuccinimide, and 1.2 mmol of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride and 10mL N,N-dimethylformamide were stirred at 25°C for 18h under the protection of helium. After the reaction was completed, the reaction mixture was divided into eight 1.5mL centrifuge tubes, centrifuged at 10000rpm for 30min, and the supernatant was divided into eight 2mL glass sealed bottles and stored under the protection of helium gas for later use.

[0065] (2) Preparation of Oxazepam Antigen (Oxazepam-Bovine Serum Albumin):

[0066] Take 100mg BSA in a 25mL single-necked bottle, add 8mL of 0.1M sodium carbonate-sodium bicarbonate buffer solution with pH 9.6 and 1.4mL N,N-dimethylformamide and stir to dissolve,...

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Abstract

The present invention relates to an oxazepam hapten, an oxazepam antigen, a preparation method and applications thereof, the structure of the oxazepam hapten is represented by a formula (I), the preparation method of the oxazepam hapten comprises that oxazepam and dianhydride are adopted as raw materials, a Friedel-Crafts acylation reaction is adopted, and carboxyl is introduced to the 5-positionbenzene ring of oxazepam, and the oxazepam hapten is obtained. The preparation method of the oxazepam antigen comprises the following steps: activating the oxazepam hapten, and carrying out a couplingreaction on the oxazepam hapten and carrier protein to obtain the oxazepam antigen. The invention initiates the preparation method of the oxazepam artificial hapten and the oxazepam artificial antigen for the first time, the synthesis steps are simple and effective, the oxazepam artificial hapten and the oxazepam artificial antigen can be completely used in immunoassay, and a convenient way is provided for people to detect and research oxazepam in the future.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an oxazepam hapten, an oxazepam antigen, a preparation method and application thereof. Background technique [0002] Oxazepam (Oxazepam) is a benzodiazepine sedative and hypnotics, alias: Shuning, chemical name: 7-chloro-2,3-dihydro-3-hydroxy-5-phenyl-1,4(2H )-benzodiazepine-2-one. Oxazepam is the main active metabolite of diazepam and belongs to the short-acting benzodiazepine class. Clinically, oxazepam is used for short-term treatment of anxiety and anxiety-related insomnia. Oxazepam has less effect on liver function and is more suitable for elderly patients and patients with liver diseases. Many patients will develop dependence after long-term use of oxazepam tablets, and withdrawal reactions will occur after stopping the drug. For this reason, many countries include these drugs in the list of prohibited drugs and strictly control them. At present, the detection of oxazepam at ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D243/24C07D243/26C07K14/765C07K14/77C07K14/795C07K14/805C07K14/47C07K16/00C07K1/107C07K1/34G01N33/558G01N33/58
CPCC07D243/24C07D243/26C07K14/47C07K14/765C07K14/77C07K14/795C07K14/805C07K16/00G01N33/558G01N33/585G01N33/9486
Inventor 袁强洪裕好黄俊兴
Owner GUANGZHOU WONDFO BIOTECH
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