Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof

A technology for rosiglitazone and artificial antigen, which is applied in the field of artificial antigen and antibody preparation, can solve the problems such as the lack of an immunological detection method for rosiglitazone, and achieves the effects of high affinity, high speed and low detection cost

Active Publication Date: 2009-09-23
华农(潮州)食品研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is no effective immunological detection method for rosiglitazone

Method used

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  • Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof
  • Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof
  • Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (1) Preparation of rosiglitazone hapten (n=1), as shown in the following reaction formula:

[0058]

[0059] Weigh 0.43g (1mmol) of rosiglitazone (purity: 98.5%) and 0.232g (2mmol) of sodium chloroacetic acid (analytical purity) into a round bottom flask, and dissolve the mixture with 5ml of absolute ethanol At the same time, 2 mmol potassium carbonate was added, and the reaction was refluxed and stirred at 60° C. During the reaction, 1 mmol potassium iodide was added as a catalyst, and the reaction was refluxed and stirred for 4 hours. The reaction mixture was filtered to obtain a yellow-brown filtrate, which was concentrated to obtain a brown-yellow oily substance. The brown-yellow oily substance is passed through a silica column chromatography silica gel column, and the eluent with a volume ratio of 8:1 chloroform / methanol eluent is collected, and the eluent is concentrated to dryness to obtain 0.09 g of rosiglitazone hapten.

[0060] (2) Preparation of rosiglitazone a...

Embodiment 2

[0068] (1) Preparation of rosiglitazone hapten (n=6), as shown in the following reaction formula:

[0069]

[0070] Weigh 0.43g (1mmol) of rosiglitazone (purity: 98.5%) and 0.76g (5mmol) of chloroheptanoic acid (analytical purity) sodium in a round bottom flask, and use 5ml of absolute ethanol for the mixture Dissolve, while adding 5mmol of potassium carbonate, reflux and stir the reaction at 50°C, add 5mmol of potassium iodide as a catalyst during the reaction, and stop the reaction after refluxing and stirring for 5 hours. The reaction mixture was filtered to obtain a yellow-brown filtrate, which was concentrated to obtain a brown-yellow oily substance. Chromatography on a silica gel column, collect the eluent with a volume ratio of 7:1 chloroform / methanol eluent, concentrate the eluent to dryness, and obtain 0.08 g of rosiglitazone hapten.

[0071] (2) Preparation of rosiglitazone artificial antigen, using active ester method to prepare immune antigen, and mixed acid anhydrid...

Embodiment 3

[0076] Example 3 Preparation of rosiglitazone hapten (n=1), R is Br, as shown in the following reaction formula:

[0077]

[0078] Weigh 0.43g (1mmol) of rosiglitazone (purity: 98.5%) and 0.16g (1mmol) of sodium bromoacetate (analytical purity) in a round bottom flask, and dissolve the mixture with 5ml of absolute ethanol At the same time, 1mmol of potassium carbonate was added, and the reaction was refluxed and stirred at 70°C. During the reaction, 0.75mmol of potassium iodide was added as a catalyst, and the reaction was stopped after refluxing and stirring for 6 hours. The reaction mixture was filtered to obtain a yellow-brown filtrate, which was concentrated to obtain a brown-yellow oily substance. Chromatography silica gel column through silica column, collect the eluent of the chloroform / methanol eluent phase with a volume ratio of 6:1, and concentrate the eluent to dryness to obtain 0.10 g of rosiglitazone hapten.

[0079] (2) Preparation of rosiglitazone artificial antig...

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Abstract

The invention discloses a rogridone hapten as well as a corresponding artificial antigen and an antibody thereof. The invention also discloses a preparation method and an application for the rogridone hapten as well as the corresponding artificial antigen and the antibody. The method comprises the following steps: adopting rogridone as a raw material which reacts with halogenated benzyl benzoate to generate the carboxyl-contained rogridone hapten; then, coupling the rogridone hapten and protein by an activated fat method or a mixed acid anhydride method to prepare the rogridone artificial antigen; and using the rogridone artificial antigen to immunize animals so as to generate the high-specificity rogridone antibody. An immunodetection method for the rogridone antibody and the rogridone artificial antigen can be applied to the on-site rapid detection of the rogridone and has an important realistic meaning to realize the rapid detection of the health food safety.

Description

Technical field [0001] The present invention relates to a rosiglitazone hapten and its corresponding artificial antigen and antibody, and also relates to the preparation method and application of the rosiglitazone hapten and its corresponding artificial antigen and antibody. Background technique [0002] Rosiglitazone (Rosiglitazone) is also known as Avandia, its chemical name is 5-[4-[2-(N-methyl-N-(2-pyridyl)amino)ethoxy]benzyl]thiazolidine -2,4-dione, molecular weight is 429.92, almost insoluble in water, its structural formula is: [0003] [0004] Rosiglitazone is a thiazolidinedione anti-diabetic drug. It is a highly selective peroxisome proliferation-activated receptor γ (PPAR-γ) agonist. It activates fat, skeletal muscle and liver by binding to PPAR-γ. PPAR-γ in the tissues that insulin acts on increases the synthesis of a variety of proteins, regulates the transcription of insulin response genes, controls the production, transport and utilization of blood sugar, correc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D417/12C07K14/765C07K14/795C07K14/77C07K14/42C07K14/435C07K16/44C12N15/06C12P21/08G01N33/64G01N33/02
Inventor 孙远明雷红涛沈玉栋石敏杨金易王弘徐晓艳
Owner 华农(潮州)食品研究院有限公司
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