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CN-121991018-A - CO fluorescent probe, preparation method thereof and application of CO fluorescent probe in reagent for detecting inflammatory reaction in organism

CN121991018ACN 121991018 ACN121991018 ACN 121991018ACN-121991018-A

Abstract

The invention relates to the field of fluorescent probes, in particular to a CO fluorescent probe, a preparation method thereof and application thereof in a reagent for detecting an inflammatory reaction in a living body. The probe uses DBX-OH with a push-pull electronic structure as a red fluorophore, and is connected with a recognition unit allyl chloroformate through a phenolic hydroxyl group of the red fluorophore. The detection mechanism is that when the fluorescent probe does not react with CO, the fluorescent probe is in an off state due to the photoinduced electron transfer effect, and when the fluorescent probe reacts with CO specifically under the catalysis of Pd (0), the allyl chloroformate chain breaks away, the PET process is inhibited, so that the strong red fluorescent light of DBX-OH is recovered, and the on detection is realized. The probe has the outstanding advantages of red light emission, excellent water solubility, extremely low detection limit and the like. And the method is successfully applied to in vivo inflammation detection of mice and zebra fish models, and the dual application potential of the method in the field of biomedical research is highlighted.

Inventors

  • Zhai Heshu
  • MENG QINGTAO
  • SHU LI
  • DENG YINJIE
  • ZHU SHIHAI

Assignees

  • 辽宁科技大学

Dates

Publication Date
20260508
Application Date
20260130

Claims (10)

  1. 1. The CO fluorescence probe is characterized in that the structural formula of the CO fluorescence probe is shown as the following formula I: Formula I.
  2. 2. The method for preparing the CO fluorescence probe according to claim 1, comprising the following steps: mixing 4- (diethylamino) benzaldehyde, 6-hydroxy-1-tetralone and an acid solution, and performing a first reaction to obtain an intermediate; and mixing the intermediate, allyl chloroformate, cesium carbonate and a solvent, and performing a second reaction to obtain the CO fluorescent probe.
  3. 3. The method for preparing a CO fluorescent probe according to claim 2, wherein the molar ratio of the 4- (diethylamino) benzaldehyde to the 6-hydroxy-1-tetralone is 1:1; the acid solution comprises aqueous solution of perchloric acid and glacial acetic acid; the volume ratio of the perchloric acid aqueous solution to the glacial acetic acid is 3:20; The concentration of the aqueous solution of perchloric acid is 70 wt%; The dosage ratio of the 4- (diethylamino) benzaldehyde to the perchloric acid aqueous solution is 10 mmol/3 mL.
  4. 4. The method for preparing a CO fluorescent probe according to claim 2, wherein the molar ratio of the intermediate, allyl chloroformate and cesium carbonate is 1:2:2; the solvent comprises acetonitrile.
  5. 5. The method for preparing a CO fluorescent probe according to claim 2, wherein the temperature of the first reaction is 90-100 ℃, and the time of the first reaction is 1.5-3 hours.
  6. 6. The method for preparing a CO fluorescent probe according to claim 2, wherein the temperature of the second reaction is 80-82 ℃, the time of the second reaction is 8-10 hours, and the atmosphere of the second reaction is inert gas.
  7. 7. The method for preparing a CO fluorescent probe according to claim 2, further comprising cooling the obtained reaction liquid to 10-30 ℃, adding a mixed solution of ethyl acetate and petroleum ether into the reaction liquid, performing solid-liquid separation, and purifying a solid product obtained by the solid-liquid separation.
  8. 8. The method for preparing a CO fluorescent probe according to claim 2, further comprising the steps of cooling the obtained reaction liquid to 10-30 ℃ after the second reaction is finished, then performing solid-liquid separation, and performing vacuum concentration and purification on the obtained solid product.
  9. 9. Use of the CO fluorescent probe of claim 1 in a reagent for detecting inflammatory reactions in a living body.
  10. 10. The application according to claim 9, characterized in that it comprises the following steps: preparing the CO fluorescent probe into a detection solution, and then using the detection solution in a reagent for detecting inflammatory reaction in a living body; Pd 2+ is also included in the detection liquid.

Description

CO fluorescent probe, preparation method thereof and application of CO fluorescent probe in reagent for detecting inflammatory reaction in organism Technical Field The invention relates to the field of fluorescent probes, in particular to a CO fluorescent probe, a preparation method thereof and application thereof in a reagent for detecting an inflammatory reaction in a living body. Background CO is a gaseous signal molecule of the inflammatory reaction in the living body. In mammals, CO can be produced by Heme Oxygenase (HOs) during endogenous heme degradation, the lipid-soluble nature of which allows it to freely cross cell membranes and bind to heme proteins. CO activates soluble guanylate cyclase (sGC) mainly through classical pathway, increases intracellular cyclic guanosine monophosphate (cGMP) level, thus mediating physiological processes such as vasodilation, and at the same time, it also regulates ion channels and inhibits cytochrome P450 enzymes and mitochondrial respiratory chain complexes. The HO-1/CO system forms a key stress response module, is induced and activated under the conditions of oxidative stress and the like, and exerts a strong cytoprotective effect through the synergistic effect of the antioxidant circulation of biliverdin/bilirubin and CO signal transmission, and the effect is particularly remarkable in inflammatory diseases. Notably, in bacterial pneumonia, pathogen metabolism can significantly increase alveolar CO concentration, making it a potential biomarker for distinguishing bacterial from viral pneumonia. Elucidating these diverse pathophysiological functions of CO in animals is highly dependent on advanced technologies that enable real-time, selective, in situ monitoring in complex living systems. The CO fluorescent probe technology has the advantages of no damage to tissues, simple operation and strong tissue penetrating power, and is suitable for real-time detection of inflammation of organisms. However, the related properties of the existing CO fluorescent probe, such as shorter emission wavelength, weak tissue penetration, longer reaction time, poor water solubility, and the like, limit the further application of the probe in living bodies. The CO fluorescent probe with good water solubility and low detection limit has remarkable advantages in-vivo application. The excellent water solubility ensures the excellent biocompatibility, can be directly used for real-time and in-situ detection in physiological environment, and avoids the interference and toxicity of organic solvents on cells or biological systems. The extremely low detection limit gives the probe extraordinary sensitivity, and can accurately capture and visualize the extremely low concentration carbon monoxide molecules in organisms, which is important for revealing the trace change of the carbon monoxide molecules in the physiological and pathological processes such as nerve signal transduction, anti-inflammation and the like. The combination of the two makes the probe a powerful tool for researching the biological function of CO on the living cell level and the living body level. Therefore, it is of great importance to provide a CO fluorescent probe that has good water solubility and a low detection limit. Disclosure of Invention The invention provides a CO fluorescent probe, a preparation method thereof and application thereof in a reagent for detecting an inflammatory reaction in a living body, wherein the CO fluorescent probe has red light emission, excellent water solubility and extremely low detection limit. In order to solve the technical problems, the invention provides the following technical scheme: The invention provides a CO fluorescence probe (ZHS-CO), wherein the structural formula of CO fluorescence is shown in the following formula I: Formula I. The mechanism of action of the CO fluorescent probe is shown in figure 1, and is based on the Pd (0) -mediated Tsuji-Trost reaction principle, wherein DBX-OH fluorophores serving as signal units in ZHS-CO are connected with allyl chloroformate serving as a recognition unit through phenolic hydroxyl groups. In the absence of reaction with carbon monoxide (CO), the probe is initially in the "off" state of fluorescence quenching due to Photoinduced Electron Transfer (PET) from the DBX-OH fluorophore (electron donor) to the allyl chloroformate moiety (electron acceptor). When the probe encounters CO, a specific reaction occurs under Pd (0) catalysis, and the allyl chloroformate chain is cleaved and detached. The key step effectively inhibits the PET process, so that the inherent strong red fluorescence of the DBX-OH fluorophore is recovered, and the sensitive 'open' -type fluorescence detection for CO is realized. The second aspect of the present invention also provides a preparation method of the CO fluorescent probe, wherein the synthetic route is shown in fig. 2, and the preparation method comprises the following steps: mixing 4- (diethylamino) benzal