A kind of preparation method of eddp artificial antigen

Abstract

The invention provides a preparation method for an artificial antigen of a methadone metabolite-2-ethylidene-1,5-dimethyl-3,3-diphenyl pyrrolidine (hereinafter referred to as EDDP). The method comprises the following steps: firstly, performing preparation and detection on a hapten, wherein diphenylacetonitrile and 2-chloro-N,N-dimethyl propylamine hydrochloride are taken as raw materials, and an addition reaction, an esterification reaction, a cyclization reaction, a nitratlon reaction and a Friedel-Crafts reaction are performed to obtain the hapten containing carboxyl; secondly, performing preparation and detection on the artificial antigen, wherein the hapten is combined with bovine gamma globulin (BGG) through a carbodiimide method to prepare the EDDP artificial antigen, namely EDDP-BGG. The prepared EDDP artificial antigen can be subjected to animal immunization to obtain a corresponding EDDP antibody, and can be used for making a research on various methadone immune analysis methods; a relatively convenient, quick and accurate way is provided for detection of methadone.

Description

technical field

[0001] The invention belongs to the technical field of biochemical engineering, and in particular relates to a method for preparing an artificial antigen of methadone metabolite (EDDP). Background technique

[0002] Methadone, also known as Mesantol, is a narcotic analgesic. Its chemical structure is far from that of morphine, but its pharmacological effects are very similar to morphine. Its prominent features are strong analgesic effect, effective oral administration, long-lasting inhibitory effect on withdrawal symptoms of opioid drug addicts, continuous and effective repeated application, and suitable for detoxification treatment of addicted patients.

[0003] The metabolic site of methadone is mainly in the liver, and the metabolic pathway is demethylation on the nitrogen atom, and then undergoes self-cyclization to obtain the main metabolite 2-acetal-1,5-dimethyl-3,3-diphenyl Dipyrrolidine (EDDP), the structural formula is as follows:

[0004] [00...

Examples

Embodiment 1

[0035] (1) Preparation of EDDP artificial hapten:

[0036] (a) Weigh 20g of diphenylacetonitrile, 26g of 2-chloro-N,N-dimethylpropylamine hydrochloride, and 20g of solid sodium hydroxide into a 150ml single-necked round-bottomed flask, and slowly heat to 100°C. Dissolve and precipitate a white solid at the same time, the solution is lemon yellow, continue to react for 6 hours; after the reaction, cool to room temperature, the reaction solution is extracted three times with 150ml ether, collect the ether phase, and extract the ether phase three times with 120ml concentration of 5% dilute hydrochloric acid solution , collect the hydrochloric acid phase, and adjust the alkali with 25% sodium hydroxide aqueous solution for the hydrochloric acid phase, until there is no more precipitation, the solution is divided into two layers, and the alkaline solution is extracted three times with 150ml ether, the ether phase is collected, and the ether phase is used Dry with anhydrous potassiu...

technical field

[0001] The invention belongs to the technical field of biochemical engineering, and in particular relates to a method for preparing an artificial antigen of methadone metabolite (EDDP). Background technique

[0002] Methadone, also known as Mesantol, is a narcotic analgesic. Its chemical structure is far from that of morphine, but its pharmacological effects are very similar to morphine. Its prominent features are strong analgesic effect, effective oral administration, long-lasting inhibitory effect on withdrawal symptoms of opioid drug addicts, continuous and effective repeated application, and suitable for detoxification treatment of addicted patients.

[0003] The metabolic site of methadone is mainly in the liver, and the metabolic pathway is demethylation on the nitrogen atom, and then undergoes self-cyclization to obtain the main metabolite 2-acetal-1,5-dimethyl-3,3-diphenyl Dipyrrolidine (EDDP), the structural formula is as follows:

[0004] [00...

Examples

Embodiment 1

[0035] (1) Preparation of EDDP artificial hapten:

[0036] (a) Weigh 20g of diphenylacetonitrile, 26g of 2-chloro-N,N-dimethylpropylamine hydrochloride, and 20g of solid sodium hydroxide into a 150ml single-necked round-bottomed flask, and slowly heat to 100°C. Dissolve and precipitate a white solid at the same time, the solution is lemon yellow, continue to react for 6 hours; after the reaction, cool to room temperature, the reaction solution is extracted three times with 150ml ether, collect the ether phase, and extract the ether phase three times with 120ml concentration of 5% dilute hydrochloric acid solution , collect the hydrochloric acid phase, and adjust the alkali with 25% sodium hydroxide aqueous solution for the hydrochloric acid phase, until there is no more precipitation, the solution is divided into two layers, and the alkaline solution is extracted three times with 150ml ether, the ether phase is collected, and the ether phase is used Dry with anhydrous potassiu...

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