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CN-122011043-A - Hypochlorous acid fluorescent probe with large Stokes shift, and preparation method and application thereof

CN122011043ACN 122011043 ACN122011043 ACN 122011043ACN-122011043-A

Abstract

The invention provides a hypochlorous acid fluorescent probe with large Stokes displacement, a preparation method and application thereof, and belongs to the technical field of hypochlorous acid fluorescent probes, wherein the hypochlorous acid fluorescent probe is a cationic iridium (III) complex and contains anions; the Stokes shift of the hypochlorous acid fluorescent probe is more than or equal to 170 nm, is favorable for reducing background interference caused by overlapping of excitation light and emission spectrum, improves detection signal to noise ratio, has quick response characteristic on HOCl, has higher detection sensitivity and excellent selectivity, can induce detectable fluorescent signal change after the hypochlorous acid fluorescent probe acts with HOCl in cells, thereby realizing in-situ detection and imaging of HOCl in cells, can be applied to a lambda-carrageenan-induced mouse arthritis model, realizes in-vivo fluorescent imaging of endogenous HOCl signals in inflammatory joint areas, and provides a sensitive and reliable imaging tool for research of HOCl related to inflammatory related diseases.

Inventors

  • GUO ZHIJUN
  • WEI PENG
  • WANG JINGYU
  • HU XINRU
  • LI MING

Assignees

  • 山东第一医科大学附属肿瘤医院(山东省肿瘤防治研究院、山东省肿瘤医院)

Dates

Publication Date
20260512
Application Date
20260205

Claims (10)

  1. 1. A hypochlorous acid fluorescent probe with large stokes shift, characterized in that the hypochlorous acid fluorescent probe has a structure shown in formula I: I is a kind of The hypochlorous acid fluorescent probe is a cationic iridium (III) complex and comprises an anti-anion; wherein the anti-anion is selected from one or more of PF 6 ⁻、BF 4 ⁻、Cl⁻、ClO 4 -, OTf-, or BPh 4 -.
  2. 2. The fluorescent probe for hypochlorous acid having a large stokes shift as claimed in claim 1, wherein the stokes shift of the fluorescent probe for hypochlorous acid is not less than 170 nm.
  3. 3. A method of preparing a hypochlorous acid fluorescent probe having a large stokes shift according to any one of claims 1-2, comprising the following synthetic route: S1, adding a tetrakis (triphenylphosphine) palladium catalyst into a reaction system containing 6-chlorophenanthridine, benzo [ b ] thiophene-2-boric acid, a solvent and alkali in an inert atmosphere, heating for reaction, extracting by an organic solvent, drying, concentrating, and purifying by silica gel column chromatography to obtain a ligand intermediate A; S2, adding IrCl 3 ·xH 2 O and the ligand intermediate A obtained in the step S1 into a mixed solvent, heating and refluxing, cooling after the reaction is finished, filtering to obtain a precipitate, washing with water and petroleum ether in sequence, and drying under vacuum to obtain a chloro-bridge iridium dimer intermediate B; and S3, adding the chlorobridge iridium intermediate B obtained in the step S2 and 4-carboxyl-4 '-methyl-2, 2' -bipyridine into a reaction vessel, adding a solvent, heating and refluxing under an inert atmosphere for reaction, then adding salt containing anions for anion exchange, and stirring at room temperature to obtain the compound of the formula I.
  4. 4. The method for preparing the fluorescent probe with large Stokes shift according to claim 3, wherein in the step S1, the equivalent ratio of tetra (triphenylphosphine) palladium to 6-chlorophenanthridine is 1 (20-50), the solvent is one or more selected from dioxane, tetrahydrofuran, toluene, dimethylformamide and ethanol/water mixed solvents, the base is one or more selected from potassium carbonate, cesium carbonate, potassium phosphate and sodium tert-butoxide, and the extraction organic solvent is one or more selected from dichloromethane, ethyl acetate and toluene.
  5. 5. The method for preparing a hypochlorous acid fluorescent probe with large Stokes shift according to claim 3, wherein in the step S2, the molar ratio of IrCl 3 ·xH 2 O to the ligand intermediate A obtained in the step S1 is 1 (1-10), and the mixed solvent is selected from glycol ether/water systems or alcohol/water systems.
  6. 6. The method for preparing a fluorescent probe for hypochlorous acid with large Stokes shift according to claim 3, wherein in the step S3, the molar ratio of the chlorobridge iridium intermediate B to 4-carboxyl-4 '-methyl-2, 2' -bipyridine is 1 (1-10), the solvent is selected from one or more of dichloromethane, chloroform, methanol, ethanol and acetonitrile, and the salt containing anions is selected from one or more of ammonium hexafluorophosphate, ammonium tetrafluoroborate, potassium hexafluorophosphate and tetraphenylborate.
  7. 7. Use of a fluorescent probe of hypochlorous acid having a large stokes shift as claimed in claim 1 for selective detection of hypochlorous acid.
  8. 8. The use according to claim 7, wherein the detection mechanism of the hypochlorous acid fluorescent probe is: After the compound with the structure of the formula I acts with HOCl, sulfur atoms in the structure of the compound of the formula I are oxidized into sulfoxide, so that the fluorescence signal of the sulfoxide is subjected to detectable change, and the compound is used for realizing qualitative or quantitative detection of HOCl.
  9. 9. Use of the hypochlorous acid fluorescent probe with large stokes shift according to claim 1 in situ detection and imaging analysis of HOCl in a model of inflammatory disease.
  10. 10. The use according to claim 9, wherein the model of inflammatory disease comprises a model associated with inflammatory joint disease, which is a model of lambda carrageenan-induced mouse arthritis.

Description

Hypochlorous acid fluorescent probe with large Stokes shift, and preparation method and application thereof Technical Field The invention belongs to the technical field of hypochlorous acid fluorescent probe detection, and particularly relates to a hypochlorous acid fluorescent probe with large Stokes displacement, and a preparation method and application thereof. Background Hypochlorous acid (HOCl) is one of the important reactive oxygen species in the organism, produced mainly by the catalysis of the myeloperoxidase system, playing a key role in the immune defenses and inflammatory response processes. Studies show that abnormal elevation of HOCl levels is closely related to inflammation-related pathological processes such as inflammatory joint diseases. Therefore, an analysis tool capable of rapidly and accurately detecting HOCl in a complex biological environment and realizing visual imaging is constructed, and the analysis tool has important significance for mechanism research and early evaluation of related diseases. At present, the fluorescent probe detection technology is widely used for detecting and imaging bioactive small molecules due to the advantages of high sensitivity, quick response, capability of realizing real-time in-situ imaging, relatively simple and convenient operation and the like. In the HOCl detection field, organic small molecular probes based on fluorophores such as coumarin, 1, 8-naphthalimide, fluorescein, rhodamine, and dipyrromethene difluoride (BODIPY) have been reported, and show a certain application value in HOCl imaging at the cellular and even animal level. However, the problem of small stokes shift of the conventional multi-class organic small molecule HOCl fluorescent probes is generally existed, which is easy to cause overlap of excitation light and emission spectrum, thereby causing background interference, signal crosstalk, fluorescence self-absorption/self-quenching and other phenomena, causing signal-to-noise reduction and limited quantitative accuracy, and being more prominent especially in deep tissues or biological systems with complex backgrounds. Based on the above reasons, development of an HOCl-responsive fluorescent probe which has both large stokes shift and high selectivity and can be suitable for complex biological environments is still a technical problem to be solved in the art. Disclosure of Invention The invention aims to solve the problems in the prior art, and provides a hypochlorous acid fluorescent probe with large Stokes displacement, a preparation method and application thereof, wherein the hypochlorous acid fluorescent probe can realize quick response and visual detection on HOCl in a complex biological environment and has the advantages of quick response, high sensitivity, good selectivity, higher biological safety and the like, thereby overcoming the problems of small Stokes displacement, obvious spectrum overlapping and background interference, insufficient signal-to-noise ratio and the like of the conventional multi-class organic micromolecule HOCl probes and improving the detection and imaging effects of HOCl. In order to achieve the above purpose, the technical scheme of the invention is as follows: One of the objects of the present invention is to provide a hypochlorous acid fluorescent probe having a large stokes shift, the hypochlorous acid fluorescent probe having a structure represented by formula I: I is a kind of The hypochlorous acid fluorescent probe is a cationic iridium (III) complex and comprises an anti-anion; Wherein the anti-anion is selected from one or more of PF 6⁻、BF4⁻、Cl⁻、ClO4 -, OTf-, or BPh 4 -. Further, the Stokes shift of the hypochlorous acid fluorescent probe is more than or equal to 170 nm. The second object of the present invention is to provide a method for preparing a hypochlorous acid fluorescent probe with large stokes shift, comprising the following synthetic route: S1, adding a tetrakis (triphenylphosphine) palladium catalyst into a reaction system containing 6-chlorophenanthridine, benzo [ b ] thiophene-2-boric acid, a solvent and alkali in an inert atmosphere, heating for reaction, extracting by an organic solvent, drying, concentrating, and purifying by silica gel column chromatography to obtain a ligand intermediate A; S2, adding IrCl 3·xH2 O and the ligand intermediate A obtained in the step S1 into a mixed solvent, heating and refluxing, cooling after the reaction is finished, filtering to obtain a precipitate, washing with water and petroleum ether in sequence, and drying under vacuum to obtain a chloro-bridge iridium dimer intermediate B; and S3, adding the chlorobridge iridium intermediate B obtained in the step S2 and 4-carboxyl-4 '-methyl-2, 2' -bipyridine into a reaction vessel, adding a solvent, heating and refluxing under an inert atmosphere for reaction, then adding salt containing anions for anion exchange, and stirring at room temperature to obtain the compound of the formula I. In the