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Kratom antigen as well as preparation method and application thereof

An antigen and hapten technology, applied in the field of kratom antigen and its preparation, can solve the problems of poor specificity, low sensitivity, and unprotected secondary amines

Pending Publication Date: 2021-09-07
HANGZHOU CLONGENE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the Chinese patent document with the publication number CN108732345A discloses a detection test paper and a preparation method for detecting mitragynine and its derivatives and metabolites in human saliva. The formation of mitragynine hemisuccinate derivatives, so that it can be coupled with bovine serum albumin to produce the whole antigen, this method can be used to prepare the whole antigen of kratom through a one-step reaction, the preparation method is simple, but the secondary amine in this method If it is not protected, two sites will appear when succinic anhydride is introduced. The coupling site is unknown and the arm length is too short. The active site is easily covered by the macromolecular carrier protein and the active group in the kratom molecule cannot be retained. Therefore, it is obtained The whole antigen has the problems of low antigenic activity, poor specificity and low sensitivity

Method used

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  • Kratom antigen as well as preparation method and application thereof
  • Kratom antigen as well as preparation method and application thereof
  • Kratom antigen as well as preparation method and application thereof

Examples

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preparation example Construction

[0061] A preparation method of kratom antigen, comprising the following steps:

[0062] Preparation of mitragynine complete antigen:

[0063] a) Protected amino groups: dissolve mitragynine in an organic solvent, stir and dissolve at room temperature, and slowly add trifluoroacetic anhydride which is 1-3 times the molar amount of mitragynine in an ice-water bath under the protection of an inert gas, and remove the ice Water bath, stirring at room temperature for 2-4 hours; after the reaction, evaporate the solvent under reduced pressure in a 40-50°C water bath to obtain a light yellow oily liquid, add 5-15ml of n-hexane to dissolve, and crystallize into a white solid in an ice-water bath. Wash the white solid with n-hexane;

[0064]

[0065] mitragynine-COCF 3

[0066]b) Hydrolysis of the ester group: Add 5-15mL of methanol and 1-3ml of water to the obtained white solid, then add 5-10 times the molar amount of lithium hydroxide, stir and dissolve and continue to react fo...

Embodiment 1

[0086] Example 1: Preparation of Kratom Antigen

[0087] Preparation of complete antigen of mitragynine (Antigen1):

[0088] a) Protected amino group

[0089] Weigh 398.5mg (1mmol) of mitragynine powder in a 150ml three-necked flask, add 60ml of dichloromethane, stir and dissolve at room temperature, and slowly add 420mg (2mmol) of trifluoroacetic anhydride dropwise under the condition of ice-water bath and nitrogen protection, The ice-water bath was removed, and stirred at room temperature for 3 hours. After the reaction, the solvent was evaporated under reduced pressure in a water bath at 45° C. to obtain a light yellow oily liquid, which was dissolved by adding 10 ml of n-hexane. It can crystallize into a white solid in an ice-water bath. 420 mg (0.86 mmol) of the white solid were washed with n-hexane.

[0090] b) Hydrolysis of the ester group Add 10 mL of methanol and 2 mL of water to the above white solid, then add 158 mg (6.88 mmol) of lithium hydroxide, stir to diss...

Embodiment 2

[0110] Example 2: Preparation of Kratom Antigen

[0111] Preparation of complete antigen of mitragynine (Antigen1):

[0112] a) Protected amino group

[0113] Weigh 398.5mg (1mmol) of mitragynine powder in a 150ml three-necked flask, add 60ml of dichloromethane, stir and dissolve at room temperature, and slowly add 210mg (1mmol) of trifluoroacetic anhydride dropwise under the condition of an ice-water bath under nitrogen protection, The ice-water bath was removed, and stirred at room temperature for 3 hours. After the reaction, the solvent was evaporated under reduced pressure in a water bath at 405° C. to obtain a light yellow oily liquid, which was dissolved by adding 10 ml of n-hexane. It can crystallize into a white solid in an ice-water bath. 239 mg (0.6 mmol) of the white solid were washed with n-hexane.

[0114] b) Hydrolysis of the ester group Add 10 mL of methanol and 2 mL of water to the above white solid, then add 110 mg (4.8 mmol) of lithium hydroxide, stir to ...

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Abstract

The invention relates to the field of biological detection, and discloses a Kratom antigen as well as a preparation method and application of the Kratom antigen, the Mitragynine complete antigen and the 7-hydroxy Mitragynine complete antigen completely reserve active groups in molecular structures, so that the Kratom antigen has relatively strong specificity and relatively high sensitivity.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a kratom antigen and its preparation method and application. Background technique [0002] Kratom, of the Rubiaceae family, is a psychoactive plant that grows in Southeast Asia, especially in Thailand and Malaysia. .Fresh or dried kratom leaves can be smoked, chewed or consumed as a tea. Low doses of kratom have a stimulating effect and have long been used by manual laborers in Southeast Asia as a stimulant to offset work fatigue; high doses of kratom have a sedative-anesthetic effect. [0003] The pharmacological function of kratom is mainly realized by the alkaloids 7-hydroxy mitragynine (formula II) and mitragynine (formula I). Although the molecular structure of these alkaloids is similar to that of hallucinogens, these substances do not have hallucinogenic activity. Instead, these alkaloids primarily interact with adrenergic and opioid receptors. Therefore, it is ofte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K1/107C12N5/20C07K16/16C07D471/14
CPCC07K14/765C07K16/16C07D471/14C07K2317/92C07K2317/94Y02P20/55
Inventor 郑智彪郑曙剑
Owner HANGZHOU CLONGENE BIOTECH
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