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Meloxicam hapten and artificial antigen as well as preparation method and application of meloxicam hapten and artificial antigen

A technology of meloxicam and artificial antigen, which is applied in chemical instruments and methods, animal/human peptides, animal/human proteins, etc., can solve the problems of limited promotion and use, complex and cumbersome processing, and long detection time, and achieve practical value High, simple preparation process, high sensitivity

Active Publication Date: 2022-04-12
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods have strong specificity and high sensitivity, the sample pretreatment is complex and cumbersome, the detection time is long, and the cost is high, so the promotion and use are limited.

Method used

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  • Meloxicam hapten and artificial antigen as well as preparation method and application of meloxicam hapten and artificial antigen
  • Meloxicam hapten and artificial antigen as well as preparation method and application of meloxicam hapten and artificial antigen
  • Meloxicam hapten and artificial antigen as well as preparation method and application of meloxicam hapten and artificial antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Preparation and characterization of embodiment 1 meloxicam hapten

[0058] 1. Preparation of meloxicam hapten

[0059] 1. Preparation of meloxicam hapten shown in formula Ⅰ

[0060] (1) Preparation of Compound A

[0061]

[0062] Weigh 14.4g of 2-aminothiazole-5-carboxylic acid in 10mL of absolute ethanol, add 5% concentrated sulfuric acid, react overnight at 50°C, cool to room temperature, pour into 100mL of saturated sodium carbonate aqueous solution, add ethyl acetate to extract, spin dry to obtain Compound A.

[0063] (2) Preparation of Compound C

[0064]

[0065] Compound A and 27g of sulfonamide methyl (compound B) were added into 10mL of xylene, refluxed at 70°C for 24h, cooled to room temperature, poured into 20mL of petroleum ether, and filtered with suction to obtain compound C.

[0066] (3) Preparation of Compound D

[0067]

[0068] Dissolve compound C in 50mL of tetrahydrofuran, add dropwise 0.5mL of 1M sodium hydroxide solution, react at 50°...

Embodiment 2

[0087] Preparation and characterization of embodiment 2 meloxicam artificial antigen

[0088] 1. Preparation and characterization of meloxicam coating agent

[0089] In the preparation methods of the immunogen and the coating source, the difference mainly lies in the type of carrier protein used. Among them, KLH is mainly used for the carrier protein of the immunogen, BSA is mainly used for the carrier protein of the coating source, and the coupling method used is the active ester method.

[0090] 1. Preparation of meloxicam coater

[0091] (1) Dissolve 19 mg of the compound represented by formula I prepared in Example 1 in 0.5 mL of DMF, add 15 mg of EDC·HCL and 8 mg of NHS, and stir for 10 hours at room temperature to obtain solution A.

[0092] (2) 20 mg of BSA was dissolved in 10 mL of PBS buffer to obtain solution B.

[0093] (3) Add solution A to solution B dropwise, stir at room temperature, and react overnight to obtain the reaction product, namely meloxicam coatogen...

Embodiment 3

[0104] The preparation of embodiment 3 meloxicam antiserum

[0105] Mix and emulsify the meloxicam immunogen prepared in Example 2 with an equal amount of Freund's adjuvant, and then immunize female Balb / c mice aged 6-8 weeks and weighing 18-20 g, using complete Freund's adjuvant for the initial immunization (purchased from Sigma Company), and incomplete Freund's adjuvant (purchased from Sigma Company) was used for booster immunization. The way of immunization was multi-point subcutaneous injection on the back of the neck, the immunization dose was 100 μg per mouse, and the immunization interval was 4 weeks, a total of three immunizations. On the seventh day after each immunization, blood was collected from the tail vein, centrifuged at 4000 rpm for 10 min, and the supernatant was collected as antiserum, which was stored at -20°C.

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Abstract

The invention provides a meloxicam hapten and an artificial antigen as well as a preparation method and application thereof. The structure of the meloxicam hapten is shown as a formula I, a formula II or a formula III, and the meloxicam artificial antigen is obtained by coupling the hapten with carrier protein. A high-specificity antibody can be obtained by immunizing animals with the meloxicam artificial antigen. The hapten, the artificial antigen and the anti-meloxicam antibody of the meloxicam are prepared for the first time, the blank at home and abroad is filled, and a new means is provided for establishing a rapid, simple, convenient, cheap, sensitive and specific meloxicam detection method.

Description

technical field [0001] The invention belongs to the technical field of biochemical engineering, and in particular relates to a meloxicam hapten and an artificial antigen and a preparation method and application thereof. Background technique [0002] Meloxicam (MLX) is a non-steroidal anti-inflammatory drug of the Xicam class, and its mechanism of action is to reduce inflammatory mediators such as prostaglandins by selectively inhibiting cyclooxygenase-2 (COX-2). Antipyretic, analgesic and anti-inflammatory effects. Meloxicam is widely used in animal husbandry, and it is commonly used clinically for acute inflammation and pain in large animals such as pigs, cattle, and horses caused by various diseases and infections; it has a significant drug synergistic effect when combined with antibiotics, and can Effectively improve the clinical symptoms of the disease. [0003] Because meloxicam has good anti-inflammatory, analgesic effects and low price, it lacks restraint and excess...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D417/12C07D279/02C07K14/765C07K14/77C07K14/795C07K14/47C07K16/44G01N33/68G01N33/577G01N33/53
Inventor 王战辉温凯余文博沈建忠米佳飞史为民
Owner CHINA AGRI UNIV