5-fluorocytosine derivative, its preparation method and its application in 5-fluorocytosine immunoassay reagent

A technology for flucytosine and derivatives, which is applied in the preparation thereof, the application of 5-fluorocytosine immunodetection reagents, and the field of 5-fluorocytosine derivatives can solve the problems of complicated operation, high analysis cost, and rising blood drug concentration. Advanced problems, to achieve the effect of overcoming complex operation, good detection effect and accurate results

Active Publication Date: 2021-07-13
长沙博源医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The combined use of 5-fluorocytosine and amphotericin B has a synergistic effect, but it can reduce the excretion of this product from the kidneys, increase the blood drug concentration, and promote the occurrence of toxic reactions in the kidney and blood system; the simultaneous use of bone marrow inhibitors can also increase the amount of this product. haematotoxicity
These methods have their unavoidable shortcomings. For example, high performance liquid chromatography and liquid chromatography-tandem mass spectrometry often have cumbersome operations, low efficiency, long measurement cycle and high analysis costs, which limit their application in clinical large-scale drug concentration. It is widely used in monitoring, while capillary electrophoresis is mostly used in scientific research laboratories, and it is difficult to meet the requirements of clinical accurate quantification
At present, there is a lack of 5-fluorocytosine detection reagents with good stability, high sensitivity and strong specificity in the market, especially high-throughput automated detection reagents with good quality

Method used

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  • 5-fluorocytosine derivative, its preparation method and its application in 5-fluorocytosine immunoassay reagent
  • 5-fluorocytosine derivative, its preparation method and its application in 5-fluorocytosine immunoassay reagent
  • 5-fluorocytosine derivative, its preparation method and its application in 5-fluorocytosine immunoassay reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1: Preparation of 5-fluorocytosine derivatives

[0073] The structural formula of 5-fluorocytosine derivatives is shown in formula (II):

[0074]

[0075] Formula (Ⅱ),

[0076] The specific preparation steps of the above-mentioned 5-fluorocytosine derivatives are as follows:

[0077] ;

[0078] (1) Synthesis of compound 3: 2.6 g of compound 1 (0.02 mmol) and 3.6 g of compound 2 (0.02 mmol) were mixed in 2.8 g of K 2 CO 3 (0.02 mmol) in 25 ml of acetonitrile (ACN) and heated to reflux for 20 hours to prepare a reaction mixture. The reaction mixture was diluted with ethyl acetate, washed with purified water and brine, dried over sodium sulfate, and finally evaporated to obtain 2.5 g of compound 3 as a white foam.

[0079]

[0080] (2) Synthesis of 5-fluorocytosine acid derivatives: 2.3 g of compound 3 (0.01 mmol) was dissolved in 20 ml of methanol (MeOH) to prepare a reaction solution, and 0.8 g of LiOH was added to the reaction solution at room temper...

Embodiment 2

[0087] Embodiment 2: Preparation of 5-fluorocytosine immunoassay reagent

[0088] The specific preparation steps of 5-fluorocytosine immunoassay reagent are as follows:

[0089] (1) Add 0.25% bovine serum albumin, 50mmol / L glucose-6-phosphate and 50mmol / L oxidized nicotinamide adenine dinucleotide to 50mmol / L Tris buffer and stir to dissolve to make R1 buffer, Add the anti-5-fluorocytosine specific antibody to the above R1 buffer at a volume ratio of 1:1500 and mix well, then adjust the pH to 8.0 with 6 mol / L hydrochloric acid to prepare the R1 reagent;

[0090] (2) Add 0.25% bovine serum albumin to 100mmol / L Tris buffer, stir and dissolve to make R2 buffer, then add 5-fluorocytosine enzyme-labeled conjugate to the above R2 buffer at a volume ratio of 1:3000 Mix well in the solution, then adjust the pH to 7.6 with 6 mol / L hydrochloric acid to make R2 reagent;

[0091] (3) Add the pure powder of 5-fluorocytosine to 6 parts of Tris-HCl buffer solution with a concentration of 5...

Embodiment 3

[0105] Example 3: 5-fluorocytosine sample detection using 5-fluorocytosine immunoassay reagent

[0106] 1. Make a standard curve:

[0107] (1) Set the reaction parameters of the Mindray BS480 automatic biochemical analyzer (Table 1).

[0108] (2) The operation steps are: first add the R1 reagent, then add the calibrator, and finally add the R2 reagent. After adding reagent R2, measure the OD340nm absorbance value at different time points, and calculate the reaction rate at different concentrations of the calibration solution. Optimal reaction standard curve plots, such as figure 1 shown.

[0109] Table 1 The reaction parameters of Mindray BS480 automatic biochemical analyzer to detect 5-fluorocytosine

[0110]

[0111] 2. Sample detection: use the standard curve obtained by the 5-fluorocytosine immunoassay reagent of the present invention to repeatedly measure low, medium and high-concentration quality control samples 10 times, and the above-mentioned quality control sa...

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Abstract

The present invention obtains a brand-new 5-fluorocytosine derivative, and uses the 5-fluorocytosine derivative to prepare a highly immunogenic 5-fluorocytosine artificial antigen, and then immunizes experimental animals to obtain a high-titer anti-antigenic antigen. 5-fluorocytosine-specific antibody; meanwhile, use the 5-fluorocytosine derivative to prepare a 5-fluorocytosine enzyme-labeled conjugate. The 5-fluorocytosine immunoassay reagent containing the above-mentioned anti-5-fluorocytosine specific antibody and 5-fluorocytosine enzyme-labeled conjugate can realize the automatic determination of 5-fluorocytosine content on a fully automatic biochemical analyzer , can high-throughput, rapid and accurate determination of 5-fluorocytosine content in biological samples, and has the advantages of simple operation, high sensitivity, strong specificity, accurate results, etc., effectively reducing the cost of 5-fluorocytosine detection, It is beneficial to widely popularize and use clinically.

Description

technical field [0001] The invention belongs to the field of biomedical testing, and specifically relates to 5-fluorocytosine derivatives, a preparation method thereof and an application in 5-fluorocytosine immunodetection reagents. Background technique [0002] 5-fluorocytosine (5-FC) is an antifungal drug, which has antibacterial effect at low concentration and bactericidal effect at high concentration. Its mechanism of action is that the drug enters the cell through the osmotic enzyme system of the fungal cell, and is converted into 5-fluorouracil under the action of the cytosine deaminase possessed by the fungal cell, and replaces uracil into the ribonucleic acid chain of the fungus, thereby blocking Normal synthesis of nucleic acids, leading to apoptosis of fungal cells. 5-Fluorocytosine is mainly used to treat fungal infectious diseases caused by Cryptococcus and Candida, such as fungal sepsis, endocarditis, meningitis, lung and genitourinary tract infections, etc. I...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D403/12C07K14/47C07K14/765C07K14/77C07K14/795C07K16/44G01N33/535G01N33/543
CPCC07D403/12C07K14/47C07K14/765C07K14/77C07K14/795C07K16/44G01N33/535G01N33/543
Inventor 张小可封飒飒成志鹏张英
Owner 长沙博源医疗科技有限公司
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