Fluorescent probe for detecting generation of nitric oxide and use thereof

Abstract

The invention relates to a fluorescent probe for measuring generation of nitric monoxide and application of the fluorescent probe. The fluorescent probe is copper complex formed by Cu<2+> and organic ligand 2-(2-hydroxy phenyl) benzimadazoles derivatives, and has the structure of a formula (I) or a formula (II). The complex can simply and quickly enter into viable cells for specific acquisition of cells or active NO in tissues, thereby the fluorescence intensity of the complex is obviously increased, consequently the complex can be applied to the detection of NO in a chemical system, analytic detection and fluorescence imaging detection of NO in viable cells or living tissues of organisms, the detection of NO in lesion tissues clinically, and real-time detection of NO of living biological systems. The fluorescent probe has high NO measurement sensitivity; the minimum lower detectable limit of the fluorescent probe is 17 nmol / L; the fluorescent probe has good selectivity on the NO; fluorescence change is quick before and after the reaction with the NO; and the fluorescence is stable. Simultaneously, the probe has good stability, can be stored for use for a long time, and is suitable for various environments of neutrality, subacidity and alkalinity.

Description

【Technical field】: [0001] The invention belongs to the technical field of biological detection and clinical medical detection, and relates to the measurement of NO in living cells or tissues, in particular to a copper complex fluorescent probe which can be used for imaging measurement of NO in living cells and its application. 【Background technique】: [0002] NO is a gas with small molecules and simple structure. It is insoluble in water and easily diffuses through cell membranes. Because there is an unpaired electron in its molecule, it has a free radical structure and is extremely unstable, and it is easy to react with oxygen, superoxide free radicals, and transition metal ions. The half-life in the tissue is extremely short, generally less than 1 min, but the half-life can be extended to about 4 min outside the tissue or under the condition of oxygen. NO can be quickly oxidized to nitrate or nitrite, combined with superoxide ions, hemoglobin and heme-containing proteins,...

AI Technical Summary

Problems solved by technology

Since 1995, laboratories around the world have devoted themselves to the development of bioluminescence detection reagents for NO, such as diaminonaphthalene and its derivatives (Diaminonaphthalene, DAN), dichlorofluorescin (DCFH), ferrous iron and cobalt ions. Complexes, diaminofluoresceins (DAFs) and diaminorhodamines (DARs), etc., but there are various defects in the current fluorescent detection reagents that hinder its further application, such as lack of specificity for NO detection (DAN and DCFH), low detection sensitivity (complexes of ferrous iron and cobalt ions), requires the participation of other reactants (typically molecular oxygen) (such as DAFs and DARs), etc., making the detection of NO in cells or in vivo progress slowly

The most successful NO fluorescent detection reagents currently used are DAF and DAR, but O 2 Although it is detected by directly reacting with NO to generate fluorescence, under certain conditions (such as cells or tissues in anaerobic environment), real-time three-dimensional space detection of NO in cells or in the body cannot be realized.

Images