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CN-121991055-A - Coumarin fluorescent probe with large Stokes displacement, preparation method and application thereof in HClO detection

CN121991055ACN 121991055 ACN121991055 ACN 121991055ACN-121991055-A

Abstract

The invention provides a coumarin fluorescent probe with large Stokes displacement, a preparation method and application thereof in biological detection, and belongs to the field of coumarin fluorescent probes. The coumarin fluorescent probe with large Stokes displacement has the following molecular structural formula: Wherein R is one of dimethylamino, nitro, methoxy and hydroxy. The coumarin fluorescent probe with large Stokes displacement can realize high-selectivity and rapid-response effective detection of endogenous or exogenous HClO of living cells on the premise of large Stokes displacement and near infrared emission, and meanwhile, the preparation raw materials of the coumarin fluorescent probe are easy to obtain, the preparation is simple, and the preparation efficiency is high.

Inventors

  • DU YANFENG
  • WANG JIANYONG
  • LI PEI
  • LI CHEN
  • LIU YINGKUN
  • CHU XUEZHI

Assignees

  • 山东探花药业有限公司

Dates

Publication Date
20260508
Application Date
20260121

Claims (10)

  1. 1. Coumarin fluorescent probe with large Stokes displacement is characterized by having the following molecular structural formula: ; wherein R is one of dimethylamino, nitro, methoxy and hydroxyl.
  2. 2. The method for preparing the coumarin fluorescent probe with large Stokes shift according to claim 1, which is characterized by comprising the following steps of primary reaction, secondary reaction and tertiary reaction; The one-time reaction method comprises the steps of contacting 7-diethylamino-4-methylcoumarin with Lawson reagent in a solvent environment, and heating to react to obtain a compound 2; the compound 2 has the following molecular structural formula: ; the secondary reaction method comprises the steps of contacting benzothiazole-2-acetonitrile with a compound 2 in the presence of Lewis acid and organic base under the protection of inert gas in a solvent environment, and reacting at room temperature to obtain a compound 3; the compound 3 has the following molecular structural formula: ; the three-time reaction method comprises the steps of contacting a compound 3 with a compound 4 in the presence of piperidine in a solvent environment under the protection of inert gas, and carrying out a heating reaction to obtain a coumarin fluorescent probe with large Stokes displacement; the compound 4 has the following molecular structural formula: ; the molecular structural formula of the compound 4 is one of dimethylamino, nitro, methoxy and hydroxyl.
  3. 3. The method for preparing the coumarin fluorescent probe with large Stokes shift according to claim 2, wherein the molar ratio of 7-diethylamino-4-methylcoumarin to Lawson reagent in the single reaction is 1:1.2-1.8.
  4. 4. The method for preparing the coumarin fluorescent probe with large Stokes shift according to claim 2, wherein in the one-time reaction, the temperature of the heating reaction is 105-115 ℃, and the reaction time of heat preservation is 5.5-6.5h.
  5. 5. The method for preparing the coumarin fluorescent probe with large Stokes shift according to claim 2, wherein the molar ratio of the compound 2 to benzothiazole-2-acetonitrile in the secondary reaction is 1:1.05-1.25.
  6. 6. The method for preparing coumarin fluorescent probe with large Stokes shift according to claim 2, wherein the secondary reaction is performed at room temperature for 10-12 hours.
  7. 7. The method for preparing a coumarin fluorescent probe with large Stokes shift according to claim 2, wherein in the secondary reaction, the Lewis acid is silver nitrate and the organic base is triethylamine.
  8. 8. The method for preparing a coumarin fluorescent probe having a large Stokes shift according to claim 2, wherein the molar ratio of the compound 3 to the compound 4 in the three reactions is 1:1.1-1.5.
  9. 9. The method for preparing the coumarin fluorescent probe with large Stokes shift according to claim 2, wherein in the three reactions, the temperature of the heating reaction is 88-92 ℃, and the reaction time is 10-12h.
  10. 10. Use of the coumarin fluorescent probe with large Stokes shift according to claim 1 or the coumarin fluorescent probe with large Stokes shift prepared by the preparation method according to any one of claims 2-9 for detecting endogenous or exogenous HClO at living cell level.

Description

Coumarin fluorescent probe with large Stokes displacement, preparation method and application thereof in HClO detection Technical Field The invention relates to the field of coumarin fluorescent probes, in particular to a coumarin fluorescent probe with large Stokes displacement, a preparation method and application thereof in HClO detection. Background As a fluorescent basic structure widely used, coumarin uses benzo-alpha-pyrone as a parent nucleus, and its original form has almost no visible fluorescence. In the prior art, the electron donating groups are introduced at the 6 and 7 positions or the electron withdrawing groups are introduced at the 3 and 4 positions, so that a molecular conjugated system can be prolonged, and the emission wavelength can be red shifted. Coumarin fluorescent probes developed based on the strategy have the advantages of high quantum yield, good light stability, easiness in modification, adjustable wavelength and the like, and have been widely used in the fields of biosensing, analytical detection, functional materials, medical diagnosis and treatment and the like. Hypochlorous acid (HClO) is one of the endogenous active oxygen, produced by the catalysis of H 2O2 and Cl - by Myeloperoxidase (MPO). Under physiological conditions, HClO exerts an antibacterial function through strong oxidation, and abnormal excessive HClO is closely related to various pathological processes such as cardiovascular diseases, arthritis, neurodegenerative diseases and the like and is confirmed to be a potential biomarker for early diagnosis of Osteoarthritis (OA). Meanwhile, HClO also exists in daily environments such as drinking water, household bleaching agents, swimming pools and the like, and the detection requirement on the HClO in the environments is wider. Aiming at the detection of HClO in living cells, the existing methods such as chemiluminescence, colorimetry, electrochemistry and the like are sensitive, but cannot meet the non-invasive real-time imaging requirement of the living cells. The reported fluorescent probe has the defects of small Stokes displacement (usually less than 50 nm), easiness in interference by ROS (reactive oxygen species), poor stability, short emission wavelength (usually less than 600 nm), incapability of detecting the spatial-temporal distribution of HClO in living cells in situ and the like, and can not realize the efficient detection of high selectivity and quick response of endogenous or exogenous HClO in living cells on the premise of large Stokes displacement and near infrared emission, and becomes a technical bottleneck of the fluorescent probe for detecting the HClO in the living cells. Furthermore, the existing preparation method of the fluorescent probe for detecting HClO in living cells also has the problems of difficult acquisition of raw materials, higher preparation difficulty and low preparation efficiency. Based on the method, the coumarin fluorescent probe with large Stokes displacement can realize the effective detection of high selectivity and quick response of endogenous or exogenous HClO of living cells on the premise of having large Stokes displacement and near infrared emission, and meanwhile, the preparation raw materials of the coumarin fluorescent probe are easy to obtain, the preparation is simple, the preparation efficiency is high, and the coumarin fluorescent probe has important technical significance and research value. Disclosure of Invention In order to solve the technical problems in the prior art, the coumarin fluorescent probe with large Stokes displacement provided by the invention can realize high-selectivity and rapid-response effective detection of endogenous or exogenous HClO of living cells on the premise of having large Stokes displacement and near infrared emission, and meanwhile, the preparation raw materials of the coumarin fluorescent probe are easy to obtain, the preparation is simple, and the preparation efficiency is high. In order to solve the technical problems, the technical scheme adopted by the invention is as follows: Coumarin fluorescent probe with large Stokes displacement has the following molecular structural formula: ; wherein R is one of dimethylamino, nitro, methoxy and hydroxyl. The preparation method of the coumarin fluorescent probe with large Stokes displacement comprises the following steps of primary reaction, secondary reaction and tertiary reaction; The one-time reaction method comprises the steps of contacting 7-diethylamino-4-methylcoumarin with Lawson reagent in a solvent environment, and heating to react to obtain a compound 2; the compound 2 has the following molecular structural formula: ; the secondary reaction method comprises the steps of contacting benzothiazole-2-acetonitrile with a compound 2 in the presence of Lewis acid and organic base under the protection of inert gas in a solvent environment, and reacting at room temperature to obtain a compound 3; the compound 3 has the followi