Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

A near-infrared fluorescent probe for detecting cytochrome p450 2C9 and its application

A cytochrome and fluorescent probe technology, applied in the field of near-infrared fluorescent probes, can solve the problems of drug side effects and increased drug exposure level, and achieve the effect of sensitive detection

Active Publication Date: 2022-07-19
XUZHOU MEDICAL UNIV
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the inhibitory effect of co-administered drugs on CYP2C9 metabolism is very likely to lead to increased drug exposure levels and induce severe drug side effects

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A near-infrared fluorescent probe for detecting cytochrome p450 2C9 and its application
  • A near-infrared fluorescent probe for detecting cytochrome p450 2C9 and its application
  • A near-infrared fluorescent probe for detecting cytochrome p450 2C9 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. 1,3-Dichloro-7-methoxy-9,9-dimethyl-2(9H)-acridone

[0035] Dissolve 61.6 mg (0.2 mmol) of 1,3-dichloro-7-hydroxy-9,9-dimethyl-2(9H)-acridone in 30 mL of acetonitrile, add 69.5 mg (0.5 mmol) of carbonic acid Potassium and 31 μL (0.5 mmol) methyl iodide were heated to reflux for 8 hours, cooled to room temperature, filtered, and acetonitrile was removed by distillation under reduced pressure. The remaining solid was dissolved in dichloromethane and separated by silica gel thin layer chromatography. The developing solvent was dichloro Methane: n-hexane=1:1 (volume ratio), 15.6 mg of orange solid was obtained as 1,3-dichloro-7-methoxy-9,9-dimethyl-2(9H)-acridone. Characterized by H NMR, C NMR and high-resolution mass spectrometry, the data are as follows: 1 HNMR (600MHz, CDCl 3 )δ7.63(s,1H),7.61(d,J=8.7Hz,1H),7.00(d,J=2.7Hz,1H),6.92(dd,J=8.7,2.7Hz,1H),3.92( s,3H),1.88(s,6H). 13 C NMR (150MHz, CDCl 3)δ173.22,163.06,147.47,140.66,139.44,136.62,135.86,134.47,1...

Embodiment 3

[0037] Example 3. Human recombinant CYP single enzyme incubation experiment

[0038] (1) Prepare CYP metabolism reaction incubation solution in advance, including pH 7.4 phosphate buffer (100mM), human recombinant CYP single enzyme, 1,3-dichloro-7-methoxy-9,9-dimethyl-2 The concentration of (9H)-acridone was 10 μM, and it was pre-incubated with shaking at 37°C for 3 minutes;

[0039] (2) Add 20 μL of 10 mM NADP to the reaction system + Solution initial reaction;

[0040] (3) After 30 minutes of reaction, 100 μL of ice acetonitrile was added, and the reaction was terminated after vigorous shaking;

[0041] (4) The samples were centrifuged at high speed for 20 minutes at 4°C and 20,000×g, and the supernatant was taken for fluorescence detection (E x =600nm, E m = 658 nm).

Embodiment 4

[0042] Example 4. Quantitative assessment of CYP2C9 activity in individual human liver microsomes from different sources

[0043] (1) A number of individual human liver microsomes (HLM) were selected and added to the reaction system for in vitro incubation experiments. Incubation system includes pH 7.4 phosphate buffer (100 mM), human liver microsomes, glucose 6-phosphate (10 mM), glucose-6-phosphate dehydrogenase (1 unit / mL), MgCl 2 (4mM), 1,3-dichloro-7-methoxy-9,9-dimethyl-2(9H)-acridone (10μM), pre-incubated with shaking at 37°C for 3 minutes;

[0044] (2) Add 20 μL of 10 mM NADP to the reaction system + initial reaction;

[0045] (3) After 30 minutes, 100 μL of ice acetonitrile was added, and the reaction was terminated after vigorous shaking;

[0046] (4) Under the conditions of 4°C, 20,000×g, centrifuge at high speed for 20 minutes, take the supernatant, and perform fluorescence detection (E x =600nm, E m =658nm), the rate of HLM-catalyzed dealkylation of 1,3-dichl...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A near-infrared fluorescent probe for detecting cytochrome P450 2C9 and its application belong to the technical field of biomedicine. The specific probe substrate can be used to measure the enzymatic activity of CYP2C9 in biological systems. The process of CYP2C9 enzyme activity determination is as follows: 1,3-dichloro-7-alkoxy-9,9-dimethyl-2(9H)-acridone was selected O The dealkylation reaction is a probe reaction, which quantitatively detects the elimination amount of the substrate 1,3-dichloro-7-alkoxy-9,9-dimethyl-2(9H)-acridone in unit time The activity of CYP2C9 enzymes in various biological samples was determined by the amount of its dealkylated metabolites. The invention can be used for quantitative assessment of CYP2C9 enzyme activity in biological samples from different sources, species and individuals and imaging of CYP2C9 in organisms, so as to realize the assessment of CYP2C9's ability to dispose of drugs. It can also be used for rapid in vitro screening of CYP2C9 activity modulators, and to evaluate the potential of drug-drug interactions due to inhibition or activation of CYP2C9 catalytic activity, such as drugs and candidate compounds.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a near-infrared fluorescent probe for detecting cytochrome P450 2C9 (CYP2C9) and its application. Background technique [0002] Cytochrome P450 enzymes are a class of heme-thiolate protein superfamily that are widely distributed in nature. In the human body, it can mediate the biotransformation of endogenous substances such as fatty acids, steroid hormones and cholesterol, and also play an important role in the metabolic clearance or biological activation of exogenous substances such as drugs, carcinogens, and environmental pollutants. [0003] CYP2C9 is a member of the cytochrome P450 superfamily, which is mainly distributed in the important metabolic organs of the human body such as the liver and intestine. According to statistics, CYP2C9 is involved in the metabolism of more than 20% of commonly used clinical drugs (more than 100 kinds), such as phenytoin, war...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64C07D219/06C09K11/06C12Q1/26
CPCG01N21/6428C07D219/06C09K11/06C12Q1/26C09K2211/1029G01N2333/90245
Inventor 马骁驰冯磊
Owner XUZHOU MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com