Near-infrared fluorescent probe substrate of COMT and application of near-infrared fluorescent probe substrate

Abstract

The invention discloses a COMT near-infrared fluorescent probe substrate and an application thereof. The probe substrate takes 7, 8-dihydroxy coumarin as a mother nucleus, conjugated double bonds areintroduced to the C3 position of 7, 8-dihydroxy coumarin to serve as a connecting group, the connecting group is connected with an electron withdrawing group EWG with hydrophilic / hydrophobic properties, and the EWG group is DMIC or TCFC or NIFC or BTC. Compared with the prior art, the probe substrate has the following advantages: (1) high specificity and sensitivity: a probe substrate can be specifically metabolized by COMT, and the product has good fluorescence emission characteristic; (2) easy high-throughput detection: the probe substrate can be used for determination on various common fluorescence microplate readers and clinical large biochemical instruments in laboratories, and 96 or 386 microwell plates can be used for batch detection; and (3) strong anti-interference capability: a methylation product of a probe substrate can emit fluorescence in a near-infrared region, has better capability of resisting autofluorescence of a biological matrix, and improves the quantification accuracy of an enzymatic activity method;.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a functional substrate and application thereof, in particular to a near-infrared fluorescent probe substrate of COMT and application thereof. Background technique [0002] Catechol-O-methyltransferase (Catechol-O-methyltransferase, COMT, EC 2.1.1.6) is an important phase II-binding metabolic enzyme in the human body, which can convert S-adenosylmethionine (S- adenosyl-L-methionine, SAM) to the catechol group of the substrate. Human COMT mainly exists in two forms, soluble COMT (S-COMT) and membrane-bound COMT (MB-COMT). S-COMT is the predominant form in peripheral tissues including liver and blood, and plays an important role in the detoxification and metabolism of exogenous catechol compounds and catechol estrogens. MB-COMT is the main form in the brain, and its physiological function is to metabolize and inactivate catecholamine neurotransmitters (such as dopamine, epinephr...

AI Technical Summary

Problems solved by technology

[0004] However, the currently commonly used COMT probe substrates have the following defects: 1. Low detection throughput: the methylated products that are catalyzed by COMT into two isomers need to be separated and enriched by means of HPLC, and the analysis process cumbersome and time-consuming; 2. Weak anti-interference ability: commonly used substrates are mostly endogenous compounds, which are easily interfered by biological matrices; 3. Poor stability: substrates are prone to oxidative degradation in the reaction system, which affects the accuracy of enzyme activity detection

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