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A kind of zopiclone artificial hapten, artificial antigen and preparation method and application thereof

An artificial hapten, zopiclone technology, applied in the direction of animal/human protein, organic chemistry, specific peptides, etc., can solve the problem of artificial hapten and artificial antigen without zopiclone, and achieve high sensitivity and specificity. , the effect of high immunogenicity

Active Publication Date: 2020-07-28
HANGZHOU TONGZHOU BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the prior art, there are no related reports on zopiclone artificial hapten and artificial antigen

Method used

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  • A kind of zopiclone artificial hapten, artificial antigen and preparation method and application thereof
  • A kind of zopiclone artificial hapten, artificial antigen and preparation method and application thereof
  • A kind of zopiclone artificial hapten, artificial antigen and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The preparation method of this implementation a kind of zopiclone artificial antigen (reaction process is as follows figure 1 ), including the following steps:

[0056] (1) Preparation of artificial hapten:

[0057] ① Weigh 500 mg (2.06 mmol) of 6-(5-amino-2-pyridyl)-6,7-dihydro-7-hydroxyl-5H-pyrrolo[3,4-b]pyrazin-5-one In a 50ml round bottom flask, add 20ml tetrahydrofuran and 5ml water to dissolve, then add 519μL (2.26mmol) of di-tert-butyl dicarbonate and 240mg (2.26mmol) of sodium carbonate, put it into a stirring bar, stir the reaction at room temperature for 3 hours, and the reaction is over , the solvent was evaporated to dryness under reduced pressure, 30ml of purified water was added, pH=3 was adjusted with 1N hydrochloric acid, and extracted twice with 30ml of ethyl acetate, the organic phase was collected, dried with anhydrous magnesium sulfate, filtered, and evaporated to dryness under reduced pressure to obtain Pale yellow oily product A548mg (1.60mmol); ...

Embodiment 2

[0138] The performance measurement of embodiment 2 zopiclone artificial antigen

[0139] (1) Identification of zopiclone artificial antigen:

[0140] Molar absorption coefficient ε: Zopiclone 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 were prepared with PBS buffer solution, which can be seen from the ultraviolet scanning diagram The maximum absorption wavelength of the zopiclone hapten is 273nm, the absorbance value is measured at 273nm, and parallel samples are made for each concentration. The formula for calculating the molar absorptivity (ie, the molar absorptivity) is: ε=absorbance value / molar concentration.

[0141]Determination of conjugate protein concentration: Prepare the concentration of 0μg / ml, 10μg / ml, 20μg / ml, 30μg / ml, 40μg / ml, 60μg / ml, 80μg / ml, 100μg / ml, 120μg / ml in PBS buffer Add 3ml of Coomassie Brilliant Blue staining solution to 1ml of bovine serum albumin solution, mix immediately, warm i...

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Abstract

The invention discloses an artificial zopiclone hapten, an artificial zopiclone antigen, a preparation method of the artificial zopiclone antigen and application of the artificial zopiclone antigen inpreparation of a zopiclone antibody. The molecular structural formula of the artificial zopiclone hapten is represented by formula (I), and the molecular structural formula of the artificial zopiclone antigen is represented by formula (II). The artificial zopiclone hapten maintains the characteristic structure of zopiclone to the greatest extent, contains active groups which can be coupled with carrier protein, and can be used as an antigenic determinant; the zopiclone antibody with high affinity, sensitivity and specificity can be prepared from the further obtained zopiclone artificial antigen in an immune manner, the valence of immune serum obtained from an immune white rabbit of New Zealand is 1:70000, and the zopiclone antibody can be used for rapid and accurate immunodetection and immunoassay of the zopiclone.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a zopiclone artificial hapten, an artificial antigen, a preparation method and application thereof. Background technique [0002] Zopiclone, as a third-generation sedative-hypnotic drug, can shorten sleep latency, increase slow-wave sleep, prolong sleep time, improve sleep quality and depth of sleep, reduce nighttime awakenings and early awakening times, and will not cause psychosis. Motility disorder. Zopiclone has anti-anxiety, anti-muscle relaxation and anti-convulsant effects. The residual effect is low in the next morning, and there is no hangover reaction after waking up. It can be used for insomnia caused by various reasons, especially for those who have difficulty falling asleep and maintaining sleep. patient. This product is absorbed quickly after oral administration, with a half-life of 3.5 to 5 hours, and has no accumulation effect after repeated adm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D487/04C07K14/765G01N33/53
Inventor 邵越水王镇
Owner HANGZHOU TONGZHOU BIOTECHNOLOGY CO LTD
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