Medetomidine hapten and preparation method thereof, medetomidine antigen and preparation method thereof, kit and medetomidine detection method

Abstract

The invention relates to the technical field of biochemical engineering, in particular to a medetomidine hapten, a preparation method of the medetomidine hapten, a medetomidine antigen, a preparation method of the medetomidine antigen, a kit and a medetomidine detection method. The medetomidine hapten is selected from a compound shown in the following structural formula: # imgabs0 #, q is any integer between 1 and 5, and R represents amido or carbonyl. The medetomidine hapten can be used for synthesizing a medetomidine antigen, so that the medetomidine hapten can be used for immunizing to generate an antibody, and adverse reaction caused by using medetomidine or hydrochloride thereof is reduced. Meanwhile, the medetomidine can be applied to rapid diagnosis products such as kits, medetomidine can be simply, conveniently and rapidly detected, and then the phenomenon of drug abuse can be effectively reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of biochemistry, and in particular, to a medetomidine hapten and a preparation method thereof, a medetomidine antigen and a preparation method thereof, a kit, and a detection method for medetomidine. Background Art

[0002] The structural formula of medetomidine is shown as follows: It is a very important pharmaceutical ingredient. As an α2 - adrenergic receptor agonist, by acting on the locus coeruleus in the central nervous system, it inhibits the release of norepinephrine, thereby exerting sedative and analgesic effects. It has the advantages of rapid onset, short action time, and weak respiratory depression effect, so it has high safety and applicability and is thus widely used in clinical practice.

[0003] In actual clinical applications, the hydrochloride form of medetomidine, namely dexmedetomidine hydrochloride, is generally used. Its molecular formula is C 13 H 16 N2·HCl, and its molecular weight is 236.7. Dexmedetomidine hydrochloride is mainly used in the tracheal intubation and mechanical ventilation processes of patients undergoing general anesthesia surgery. It can not only relieve the pain and anxiety of patients, but also effectively maintain the stability of perioperative hemodynamics and reduce the stress response caused by operations such as tracheal intubation.

[0004] Although dexmedetomidine hydrochloride has important medical value, there are also certain potential hazards during its use. For example, rapid injection of this drug may cause adverse reactions such as hypotension, hypertension, bradycardia, and sinus arrest. At the same time, long - term or excessive use of dexmedetomidine hydrochloride may also lead to addiction.

[0005] Therefore, in order to ensure the safe use of dexmedetomidine hydrochloride, effective detection means are needed to monitor its content and purity. Currently, a commonly used detection method is liquid chromatography, but this method has disadvantages such as long detection time and complex procedures, and it cannot achieve rapid detection to determine whether the detector is addicted.

[0006] In view of this, the present invention is specifically proposed. Summary of the Invention

[0007] The purpose of the present invention is to provide a medetomidine hapten and a preparation method thereof, a medetomidine antigen and a preparation method thereof, a kit, and a detection method for medetomidine. An embodiment of the present invention provides a medetomidine hapten, which can be used to synthesize a medetomidine antigen, and then can generate antibodies through immunization, reducing the adverse reactions caused by the use of medetomidine or its hydrochloride. At the same time, it can be applied to a kit, and then can simply and rapidly detect medetomidine, thereby effectively reducing the phenomenon of drug abuse.

[0008] The present invention is implemented as follows:

[0009] In a first aspect, the present invention provides a metomidate hapten, which is selected from the compounds represented by the following structural formulas:

[0010] Wherein, q is any integer between 1 and 5, and R represents an amino group or a carbonyl group.

[0011] In an alternative embodiment, it is selected from any one of the compounds represented by the following structural formulas: Wherein, p represents any integer between 1 and 3, and n represents any integer between 1 and 4.

[0012] In an alternative embodiment, it is selected from any one of the compounds represented by the following structural formulas:

[0013]

[0014] In a second aspect, the present invention provides a method for preparing the metomidate hapten described in the foregoing embodiment. When R is an amino group, the synthesis is carried out with reference to the following synthetic route:

[0015] Wherein, X represents a halogen, and q is any integer between 1 and 5;

[0016] When R represents a carbonyl group, the synthesis is carried out with reference to the following synthetic route:

[0017] q is any integer between 1 and 5.

[0018] In an alternative embodiment, it includes: mixing compound 1, a basic material, and an amino acid ester for reaction to form compound 2, and then carrying out a hydrolysis reaction, wherein the molar ratio of compound 1 to the amino acid ester is 1:(1.0 - 1.5); the molar ratio of compound 1 to the basic material is 1:(1.2 - 1.5);

[0019] Preferably, the basic material is selected from metal hydrides, preferably sodium hydride;

[0020] Preferably, it includes: mixing metomidate and an acid anhydride compound and reacting at 50 - 60 °C, wherein the molar ratio of metomidate to the acid anhydride compound is 1:(1.2 - 1.8).

[0021] In a third aspect, the present invention provides a metomidate antigen, and its structural formula is as follows: Wherein, q is any integer between 1 and 5, and R represents an amino group or a carbonyl group.

[0022] In an alternative embodiment, the protein comprises a carrier protein, preferably any one of BSA, KLH, BGG, and BTG.

[0023] In a fourth aspect, the present invention provides a method for preparing the meptazinol antigen described in the foregoing embodiments, comprising: activating the meptazinol hapten described in the foregoing embodiments, and then coupling it with a protein.

[0024] In a fifth aspect, the present invention provides a kit comprising the meptazinol antigen described in the foregoing embodiments.

[0025] In a sixth aspect, the present invention provides a method for detecting meptazinol, comprising: performing qualitative and / or quantitative detection using the kit described in the foregoing embodiments.

[0026] The present invention has the following beneficial effects: The embodiments of the present invention provide a meptazinol hapten, which can be used to synthesize a meptazinol antigen. This meptazinol antigen has a high immune titer and excellent antibody affinity, and can be used for immunization, thereby generating antibodies in the patient's body to reduce adverse reactions such as hypotension, hypertension, bradycardia, and sinus arrest caused during the use of meptazinol or its hydrochloride. At the same time, this meptazinol antigen can be used to prepare a kit for rapid and simple detection of meptazinol, thereby effectively preventing drug abuse. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] To more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for use in the embodiments. It should be understood that the following drawings only show some embodiments of the present invention, and therefore should not be regarded as limiting the scope. For those of ordinary skill in the art, without creative efforts, other relevant drawings can also be obtained based on these drawings.

[0028] Figure 1 Mass spectrometry diagram of hapten B provided in Example 2 of the present invention;

[0029] Figure 2 Liquid chromatography diagram of hapten B provided in Example 2 of the present invention;

[0030] Figure 3 Mass spectrometry diagram of hapten A provided in Example 1 of the present invention;

[0031] Figure 4 Liquid chromatography diagram of hapten A provided in Example 1 of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0032] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. For those not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are followed. For reagents or instruments without indicating the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0033] An embodiment of the present invention provides a metomidate hapten, which is selected from the compounds shown in the following structural formulas:

[0034] Among them, q is any integer between 1 and 5, and R represents an amino group or a carbonyl group. For example, q is any value between 1 and 5 such as 1, 2, 3, 4, and 5.

[0035] Only by using the above-mentioned metomidate hapten provided by the embodiments of the present invention can a metomidate antigen with high immune titer and excellent antibody affinity be formed. If haptens with other structures are used, the antigens formed have low immune titer and insufficient antibody affinity, and cannot be industrially and practically applied to the kit detection of metomidate or cannot produce effective antibodies.

[0036] Specifically, the metomidate hapten is selected from any one of the compounds shown in the following structural formulas: Among them, p represents any integer between 1 and 3, and n represents any integer between 1 and 4. For example, p is any one of the values 1, 2, and 3, and n is any one of the values 1, 2, 3, and 4.

[0037] For example, it is selected from any one of the compounds shown in the following structural formulas:

[0038]

[0039] In a second aspect, the present invention provides a preparation method for the metomidate hapten described in the foregoing embodiment, including the following steps:

[0040] When R is an amino group, the synthesis is carried out with reference to the following synthetic route:

[0041] Among them, X represents a halogen, for example, any one of halogens such as bromine and chlorine; q is any integer between 1 and 5.

[0042] Specifically, S1. Synthesize compound 1:

[0043] React metomidate and a halogen (such as bromine) in a molar ratio of 1:(1.2 - 2.0). Specifically, dissolve metomidate in an organic solvent (such as glacial acetic acid), then cool it to 4 - 10 °C, and then add the halogen. After adding the halogen, warm it to room temperature (20 - 30 °C) for reaction.

[0044] S2. Synthesize Compound 2;

[0045] Mix Compound 1, a basic material (such as selected from metal hydrides, preferably sodium hydride;) and an amino acid ester (such as ethyl glycinate) for reaction. Among them, the molar ratio of Compound 1 to the amino acid ester is 1:(1.0 - 1.5); for example, any value between 1:(1.0 - 1.5) such as 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4, and 1:1.5 or the range value between any two values. The molar ratio of Compound 1 to the basic material is 1:(1.2 - 1.5); for example, any value between 1:(1.2 - 1.5) such as 1:1.2, 1:1.3, 1:1.4, and 1:1.5 or the range value between any two values.

[0046] Specifically, in an amino acid ester and a solvent (such as DMF), under the protection of an inert gas, after cooling to 4 - 10 °C, add the basic material and continue stirring, and then add Compound 1, and react at room temperature (20 - 30 °C).

[0047] S3. Hydrolysis;

[0048] Mix Compound 2 and a basic substance (such as a hydroxide, including but not limited to sodium hydroxide) for reaction. Among them, the molar ratio of Compound 2 to the basic substance is 1:(8 - 20), for example, any value between 1:(8 - 20) such as 1:8, 1:10, 1:15, and 1:20.

[0049] Or when R represents a carbonyl group, refer to the following synthetic route for synthesis:

[0050] q is any integer between 1 and 5.

[0051] Specifically, mix metomidate and an acid anhydride compound (such as succinic anhydride) and react under the condition of 50 - 60 °C. Among them, the molar ratio of metomidate to the acid anhydride compound is 1:(1.2 - 1.8), for example, any value between 1:(1.2 - 1.8) such as 1:1.2, 1:1.4, 1:1.5, 1:1.7, and 1:1.8 or the range value between any two values.

[0052] It should be noted that after each of the above reactions, post-treatment will be carried out, and the post-treatment method used is a conventional post-treatment method, which will not be elaborated in the embodiments of the present invention.

[0053] In the third aspect, the present invention provides a metomidate antigen, and its structural formula is as follows: Among them, q is any integer between 1 and 5, and R represents an amino group or a carbonyl group. This meptazinol antigen has excellent immunogenicity and antibody affinity, and can then be used in an immune reaction to form antibodies to reduce adverse reactions such as hypotension, hypertension, bradycardia, and sinus arrest caused during the use of meptazinol or its hydrochloride. It can also be used to prepare a detection kit, and then can quickly and simply detect meptazinol, and can effectively prevent drug abuse.

[0054] Specifically, the protein includes a carrier protein, such as any one of BSA, KLH, BGG, and BTG, including but not limited to.

[0055] In a fourth aspect, the present invention provides a method for preparing the meptazinol antigen described in the foregoing embodiments, including: activating the meptazinol hapten described in the foregoing embodiments, and then coupling with a protein.

[0056] Among them, the steps and conditions for activating the above-mentioned meptazinol hapten are conventional operations, and will not be described in detail in the embodiments of the present invention. For example, NHS and / or EDC are used to activate the meptazinol hapten, and then coupled with a protein.

[0057] In a fifth aspect, the present invention provides a kit, which includes the meptazinol antigen described in the foregoing embodiments.

[0058] In a sixth aspect, the present invention provides a method for detecting meptazinol, including: performing qualitative and / or quantitative detection using the kit described in the foregoing embodiments.

[0059] The features and properties of the present invention will be further described in detail below with reference to the embodiments.

[0060] Example 1

[0061] This example provides a meptazinol hapten (hereinafter referred to as hapten A), and its structural formula is as follows:

[0062] This example provides a method for preparing the above-mentioned hapten A, including: synthesizing according to the following synthetic route:

[0063] The specific process is as follows:

[0064] S1. Synthesize compound 1;

[0065] 500 mg of metomidate was dissolved in 5 ml of glacial acetic acid. The temperature was lowered to 10 °C in an ice-water bath, and 360 mg of bromine was added dropwise. After the addition was complete, the ice-water bath was removed, and the temperature was slowly raised to room temperature. The reaction was continued for two hours. Work-up: The solvent was removed by concentration under reduced pressure. The residue was dissolved in 20 ml of DCM, and then slowly washed once with saturated sodium bicarbonate solution. The organic phase was separated and washed once with water, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated to dryness to obtain 700 mg of Compound 1.

[0066] S2. Synthesis of Compound 2;

[0067] 200 mg of ethyl glycinate hydrochloride was dissolved in 10 ml of DMF. Under nitrogen protection and cooling in an ice-water bath, 64 mg of NaH was added. After the addition, the reaction was stirred for an additional 30 minutes, and then 500 mg of Compound 1 was added. The reaction was carried out overnight at room temperature. Work-up: The organic solvent was removed by concentration. Then it was dissolved in 20 ml of DCM, and the organic phase was washed three times with 10 ml of water, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 400 mg of Compound 2.

[0068] S3. Synthesis of hapten A;

[0069] 300 mg of Product 2 was dissolved in 6 ml of methanol and stirred magnetically for 10 minutes. After dissolution, 4 ml of 4 mol / l NaOH was added. After the addition, the reaction was carried out overnight at room temperature. Work-up: The temperature was lowered in an ice-water bath, and the pH of the reaction solution was adjusted to 3 with dilute hydrochloric acid. It was concentrated under reduced pressure in a 40 °C water bath to remove methanol. Then it was extracted three times with 10 ml of DCM. The organic phases were combined, washed once with 10 ml of water, and the organic phase was dried over 5 g of anhydrous sodium sulfate, filtered, and the organic phase was purified by column chromatography with silica gel. The eluent ratio was DCM:MeOH = 50:1, and 210 mg of hapten A was obtained after purification.

[0070] Example 2

[0071] This example provides a metomidate hapten (hereinafter referred to as hapten B), and its structural formula is as follows:

[0072] This example provides a preparation method of the above hapten B, including: synthesizing according to the following synthetic route:

[0073]

[0074] Dissolve 300 mg of metomidate in 10 ml of pyridine, then add 180 mg of succinic anhydride, heat up to 55 °C, and react overnight. Post-treatment: Concentrate under reduced pressure in a 50 °C water bath to remove the solvent, add 10 ml of ethanol to dissolve, concentrate to dryness again, dissolve the residue in 20 ml of DCM, add silica gel for sample mixing and purify by column chromatography. The eluent ratio is DCM:MeOH = 30:1; 350 mg of hapten B is obtained.

[0075] Comparative Example 1

[0076] This comparative example provides a metomidate hapten (hereinafter referred to as hapten C), and its structural formula is as follows:

[0077] This comparative example provides a synthesis method of the above hapten C, including synthesizing according to the following synthetic route:

[0078] The specific process is as follows:

[0079] S1. Synthesize compound 3;

[0080] Weigh 200 mg of metomidate and add it to a 100 ml dry two-necked flask. At room temperature, add 10 ml of DMF, stir magnetically to dissolve the raw materials, protect with nitrogen, then add 243 mg of SeCO3, and dropwise add 234 mg of ethyl 4-bromobutyrate while stirring. After the addition is complete, heat up to 80 °C and react overnight; Post-treatment: Cool the reaction solution in an ice-water bath, then concentrate the reaction solution under reduced pressure in a 60 °C water bath to remove DMF. The residue is dissolved in 20 ml of DCM, add 30 ml of water, wash three times with 10 ml of DCM, separate and combine the organic phases. The organic phase is dried over 5 g of anhydrous sodium sulfate, filtered, and concentrated to obtain 350 mg of compound 3.

[0081] S2. Synthesize hapten C;

[0082] Take 300 mg of compound 3 and dissolve it in 10 ml of methanol, stir magnetically for 10 minutes. After dissolution, add 6 ml of 4 mol / l NaOH. After adding, react overnight at room temperature. Post-treatment: Cool in an ice-water bath, add dilute hydrochloric acid to adjust the pH of the reaction solution to 3, concentrate under reduced pressure in a 40 °C water bath to remove methanol, then extract three times with 10 ml of DCM, combine the organic phases, wash once with 10 ml of sodium chloride aqueous solution, dry the organic phase over 5 g of anhydrous sodium sulfate, filter, add silica gel to the organic phase for sample mixing and purify by column chromatography. The eluent ratio is DCM:MeOH = 30:1, and 230 mg of hapten C is obtained after purification.

[0083] Comparative Example 2

[0084] This comparative example provides a metomidate hapten (hereinafter referred to as hapten D), and its structural formula is as follows:

[0085] This comparative example provides a synthesis method of the above hapten D, including synthesis according to the following synthesis route:

[0086]

[0087] The specific operations are as follows:

[0088] S1. Synthesize compound 4;

[0089] Take 500 mg of metomidate and dissolve it in 10 ml of DCM. After stirring until clear, under nitrogen protection, cool it to about 0 °C in an ice-water bath, add 280 mg of DIPEA and 570 mg of TFAA, and stir at room temperature overnight. Post-treatment: Add 30 ml of pure water for washing, then add 10 ml of DCM for extraction three times, combine the organic phases, add 20 ml of sodium chloride aqueous solution for washing, collect the organic phase, add 5 g of anhydrous sodium sulfate for drying, filter, and concentrate to obtain 580 mg of compound 4.

[0090] S2. Synthesize hapten D;

[0091] Take 500 mg of compound 4 and dissolve it in 20 ml of DCM. After stirring until clear, under nitrogen protection, cool it in an ice-water bath, add 400 mg of anhydrous aluminum chloride, and then add 274 mg of succinic anhydride. After adding the materials, react at room temperature overnight. Post-treatment: Control the temperature in an ice-water bath, add 5 ml of 1 mol / l dilute hydrochloric acid to the reaction, filter, take the filtrate, wash it twice with water, dry it, mix it with silica gel and purify it by column chromatography. The eluent ratio is DCM:CH3OH = 20:1, and concentrate to dryness to obtain 520 mg of hapten D.

[0092] Synthesis of a series of antigens

[0093] Synthesize artificial antigen A-BSA according to the following steps:

[0094] Take 50 mg of hapten A prepared in Example 1 and dissolve it in 2 ml of DMF, then add 30 mg of NHS and 50 mg of EDC, and react at room temperature overnight. Then weigh 500 mg of BSA and dissolve it in 50 ml of PBS until evenly dissolved. Then add the reaction solution to the prepared protein solution and react at room temperature for 8 hours. Then place the solution in a dialysis bag and dialyze it in PBS buffer. During this period, replace the PBS buffer every 2 hours, and a total of 8 replacements are required. After the reaction is completed, centrifuge at 10000 r / min for 10 min to obtain the supernatant. Measure the concentration by the Lorwy method to obtain the final synthesized antigen, artificial antigen A-BSA.

[0095] Compound characterization experiment

[0096] The hapten A of metopimidine prepared in Example 1 was detected by high performance liquid chromatography (HPLC). The detection conditions were as follows: The hapten A was detected by HPLC. The detection conditions were as follows: Detection wavelength: 254 nm; Mobile phase: acetonitrile / water, where gradient elution was performed (time program: 0.01 - 5 min, 10% - 100% acetonitrile / water; 5.01 - 12 min, 100% acetonitrile); The chromatographic column was C18.

[0097] The hapten A of metopimidine prepared in Example 1 was detected by mass spectrometry.

[0098] The results are shown in Figure 3 and Figure 4 According to Figure 3 and Figure 4 it was known that the measured value of m / z [M + H] was 274.3, which was consistent with the theoretical value. According to Figure 4 it was known that the peak emergence position was at 9.983 min and the purity was above 95%. It indicated that the compound synthesized in Example 1 of the present invention was the hapten A of metopimidine with the required structure.

[0099] The hapten B of metopimidine prepared in Example 2 was detected by HPLC. The detection conditions were as follows: The hapten B was detected by HPLC. The detection conditions were as follows: Detection wavelength: 214 nm; Mobile phase: acetonitrile / water, where gradient elution was performed (time program: 0.01 - 5 min, 10% - 100% acetonitrile / water; 5.01 - 12 min, 100% acetonitrile); The chromatographic column was C18.

[0100] The hapten B of metopimidine prepared in Example 2 was detected by mass spectrometry.

[0101] The results are shown in Figure 1 and Figure 2 According to Figure 1 and Figure 2 it was known that the measured value of m / z [M - H] was 299.1, which was consistent with the theoretical value. According to Figure 2 it was known that the peak emergence position was at 8.018 min and the purity was above 95%. It indicated that the compound synthesized in Example 2 of the present invention was the hapten B of metopimidine with the required structure.

[0102] Experiment for determining molar extinction coefficient, coupling ratio and conjugate protein concentration

[0103] The hapten A of Example 1, the hapten B of Example 2, the hapten C of Comparative Example 1 and the hapten D of Comparative Example 2 were combined with different proteins to synthesize corresponding artificial antigens according to the above synthesis steps, and the molar extinction coefficient, coupling ratio and conjugate protein concentration of each artificial antigen were determined. The determination results are shown in Table 1.

[0104] Specifically, the measurement method:

[0105] Molar absorption coefficient ε: Prepare metomidate artificial hapten solutions with concentrations of 0 μg / ml, 5 μg / ml, 10 μg / ml, 20 μg / ml, 30 μg / ml, and 40 μg / ml using PBS buffer. From the UV scanning graph, it can be seen that the maximum absorption wavelength of the metomidate hapten is 242 nm. Use PBS as a control sample to measure the absorbance at 242 nm, and make parallel samples for each concentration. The calculation formula for the molar absorptivity (i.e., molar absorption coefficient) is: ε = absorbance / molar concentration.

[0106] Absorbance measurement: Prepare a 100 μg / ml bovine serum albumin PBS solution, dilute the conjugate (i.e., metomidate artificial antigen) to 100 μg / ml with PBS, and measure the absorbance values A2 and A1 at 255 nm using PBS as a blank control.

[0107] The coupling ratio γ is: γ = [(A1 - A2) / ε] / (100 × 10 3 / 68000).

[0108] Where ε is the molar absorption coefficient (L / mol), 68000 is the molecular weight of bovine serum albumin, and 100 × 10 3 is the bovine serum albumin concentration (g / L).

[0109] Determination of the conjugate protein concentration: Prepare 1 ml of bovine serum albumin solutions with concentrations of 0 μg / ml, 10 μg / ml, 20 μg / ml, 30 μg / ml, 40 μg / ml, 60 μg / ml, 80 μg / ml, and 100 μg / ml using PBS buffer, add 3 ml of Coomassie Brilliant Blue staining solution, mix immediately, warm in a water bath at 30 °C for 5 minutes, make parallel samples for each concentration, measure the absorbance at 655 nm, and plot the relationship curve between protein concentration and absorbance. Dilute the artificial antigen solution (prepared with PBS buffer) in a certain proportion and measure the absorbance of the artificial antigen at 655 nm, and read the corresponding protein concentration value of the artificial antigen solution from the curve.

[0110] Table 1 Test results of the molar absorption coefficient, coupling ratio, and conjugate protein concentration of each artificial antigen

[0111]

[0112] According to Table 1, it can be seen that each hapten small molecule is conjugated to the carrier protein to form a synthetic antigen.

[0113] Animal immunization experiment

[0114] The artificial antigens formed from the meptazinol haptens of Examples 1-2 and Comparative Examples 1-2 were respectively added with adjuvants and injected into mice for immunization. A booster injection was given after seven days, the third immunization was carried out on the 20th day, and blood was collected after one month to detect the serum titer. The results are shown in Table 2.

[0115] Table 2 Immunoserum Titer

[0116]

[0117]

[0118] As can be seen from Table 2, (1) the artificial antigens formed from hapten A and hapten B provided in the examples of the present invention have good immunization effects, and the immunization titers all meet the requirements. Further cell fusion is carried out, and through ELISA screening and then purification, a suitable antibody is finally obtained to match the antigen, which can be used in the immunoassay of meptazinol, providing a more rapid and accurate way for the detection of meptazinol.

[0119] (2) The antigens formed from the meptazinol haptens provided in the examples of the present invention have good immunization titers. If the structure of the meptazinol hapten is changed, the titer of the formed antigen is low and cannot meet the requirements of practical applications.

[0120] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A metoclopramide hapten, characterized in that: It is selected from the compounds shown in the following structural formula: Wherein, q is any integer between 1 and 5, and R represents an amine group or a carbonyl group.

2. The metoclopramide hapten according to claim 1, characterized in that It is selected from any one of the compounds shown in the following structural formulas: Wherein, p represents any integer between 1 and 3, and n represents any integer between 1 and 4.

3. The metoclopramide hapten according to claim 1 or 2, characterized in that It is selected from any one of the compounds shown in the following structural formulas:

4. A method for preparing the metoclopramide hapten according to claim 1, characterized in that: When R is an amine group, the synthesis is carried out according to the following synthesis route: Wherein, X represents a halogen, and q is any integer between 1 and 5; When R represents a carbonyl group, the synthesis is carried out according to the following synthesis route: q is any integer between 1 and 5.

5. The preparation method according to claim 4, characterized in that: include: Compound 1, an alkaline material and an amino acid ester are mixed and reacted to form compound 2, and then a hydrolysis reaction is performed, wherein the molar ratio of the compound 1 to the amino acid ester is 1:(1.0-1.5); the molar ratio of the compound 1 to the alkaline material is 1:(1.2-1.5); Preferably, the alkaline material is selected from metal hydrides, preferably sodium hydride; Preferably, the method comprises: mixing metoclopramide and an acid anhydride compound and reacting them at 50-60° C., wherein the molar ratio of the metoclopramide to the acid anhydride compound is 1:(1.2-1.8).

6. A metoclopramide antigen, characterized in that: Its structural formula is as follows: Wherein, q is any integer between 1 and 5, and R represents an amine group or a carbonyl group.

7. The metoclopramide antigen according to claim 6, characterized in that The protein includes a carrier protein, and is preferably any one of BSA, KLH, BGG and BTG.

8. A method for preparing the metoclopramide antigen according to claim 6, characterized in that: include: The metoclopramide hapten according to claim 1 is activated and then coupled to a protein.

9. A kit, characterized in that: It comprises the metoclopramide antigen according to claim 6.

10. A method for detecting metoclopramide, characterized in that: include: The kit according to claim 9 is used for qualitative and / or quantitative detection.

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