CN-122010835-A - HSO is detected to target lysosomal assay3-AIE fluorescent probe of (2)

CN122010835ACN 122010835 ACN122010835 ACN 122010835ACN-122010835-A

Abstract

The invention provides an aggregation-induced emission (AIE) fluorescent probe (Z) -3- [4- (bis {4- [ 2-morpholinoethoxy ] phenyl } amino) phenyl ] -2- (4-quinolinyl) acrylonitrile for targeting a lysosome to detect HSO 3 ‑ , namely a fluorescent probe MoTAQ, a preparation method thereof and application thereof in detection. The weak alkalinity of morpholine ring groups in the probe and weak acidity in a cell lysosome are mutually attracted to realize lysosome targeting, the molecular structure of the fluorescent probe is modified to enable the molecular structure to realize specific detection of HSO 3 ‑ , and meanwhile, the fluorescent groups in the fluorescent probe are blue-shifted from red light to blue light before and after detecting HSO 3 ‑ , so that effective observation can be carried out through color change during fluorescent detection, and secondly, moTAQ has good fluorescent emission spectrum characteristics, the fluorescent probe has high signal to noise ratio in vivo and in vitro, and the fluorescent probe also has good biocompatibility.

Inventors

XUE KE
LI CHANGQING
WAN WENJUN
WU QI
ZHOU YU
CHEN YUNYI
XU YIFAN

Assignees

盐城师范学院

Dates

Publication Date: 20260512
Application Date: 20260201

Claims (7)

1. An aggregation-induced emission (AIE) fluorescent probe for targeting lysosomes to detect hydrogen sulfite (HSO 3 - ) is characterized in that the fluorescent probe has a chemical structural formula: 。
2. The AIE fluorescent probe for targeting the lysosome to detect the HSO 3 - according to claim 1, wherein the detection process of the fluorescent probe on the HSO 3 - in vitro cells is that the fluorescent probe is dissolved in PBS buffer solution containing the HSO 3 - , wherein the pH value of the PBS buffer solution is 7.4-7.5, the temperature of the solution is 37 ℃, the concentration of the fluorescent probe in the PBS buffer solution is 10 mu M, the living cells of the cells HepG2 are incubated with a culture medium containing the fluorescent probe, and the color change generated by the reaction of the fluorescent probe and the HSO 3 - in the cells after 10 minutes is measured as an evaluation index of the concentration of the HSO 3 - .
3. The method for preparing the AIE fluorescent probe for targeting the lysosome to detect the HSO 3 - , which is characterized in that 4-bis (4-hydroxyphenyl) aminobenzaldehyde (TTBH), 4- (2-chloroethyl) morpholine, potassium iodide and cesium carbonate are placed in a reaction bottle, acetonitrile is added into the reaction bottle, the mixed material is heated to not lower than 85 ℃ under the protection of nitrogen, the reacted reaction liquid is cooled to room temperature, the organic phases are extracted and combined, 2- (quinolin-4-yl) acetonitrile and piperidine are added into the reaction bottle after the mixed reaction liquid is distilled, ethanol is added into the reaction bottle, the mixed material is heated to not lower than 85 ℃ under the protection of nitrogen, the reacted reaction liquid is cooled to room temperature, the organic phases are extracted and combined, drying and reduced pressure concentration treatment are carried out, a crude product is obtained, dichloromethane/methanol is used as an eluent, and the crude product is purified by a column chromatography method, so that the fluorescent probe (Z) -3- [4- (bis {4- [ 2-morpholinoethoxy ] phenyl } phenyl ] -2- (4-quinolinyl) acrylonitrile MoTAQ is obtained.
4. The method for preparing the AIE fluorescent probe for targeting the lysosome for detecting HSO 3 - , according to claim 6, wherein the mixing molar ratio of 4-bis (4-hydroxyphenyl) aminobenzaldehyde (TTBH) to 4- (2-chloroethyl) morpholine is 1:1-1:10.
5. The method for preparing the AIE fluorescent probe for targeting the lysosome for detecting HSO 3 - , according to claim 6, wherein the mixing molar ratio of 4-bis (4-hydroxyphenyl) aminobenzaldehyde (TTBH) to 2- (quinolin-4-yl) acetonitrile is 1:0.5-1:10.
6. The method for preparing the lysosome targeted aggregation-induced emission ionic probe according to claim 6, wherein the method comprises the following steps: the reaction time is at least 1h.
7. The method for preparing the lysosome targeted aggregation-induced emission ionic probe according to claim 6, wherein the feeding ratio of dichloromethane/methanol as an eluent is 1:1-100:1.

Description

AIE fluorescent probe for targeting lysosome to detect HSO 3- Technical Field The invention relates to the field of organic fluorescent probe molecules, in particular to a preparation method and application of an AIE fluorescent probe for detecting HSO 3- by targeting lysosomes. Background The lysosome is used as a highly dynamic acid digestion center in eukaryotic cells, is not only responsible for degradation recovery and metabolic steady state regulation of biomacromolecules, but also participates in cell fate decision through regulating and controlling ion balance, signal transduction and other ways. HSO 3- in the cavity is used as a key sulfur-containing metabolic intermediate to directly influence lysosomal enzyme activity, membrane permeability and autophagy-apoptosis balance, and abnormal concentration fluctuation of the HSO 3- is proved to be closely related to pathological processes such as lysosomal storage diseases, neurodegenerative diseases (such as Alzheimer disease and Huntington disease), tumor drug resistance and immune metabolism disorder. Because of the unique acidic microenvironment (pH 4.5-5.5) and the high-activity hydrolase system of the lysosome, the traditional detection probe is easy to generate protonation fluorescence quenching or enzymatic degradation, and is difficult to realize in-situ accurate quantification of the HSO 3-. Therefore, development of a novel detection tool with lysosome targeting specificity, acidic environment stability and dynamic response sensitivity has urgent need for revealing the molecular mechanism of lysosome-related diseases and pushing targeting therapeutic strategies. In the field of lysosome microenvironment probe design, the prior art generally faces three challenges, namely that firstly, a conventional probe lacks lysosome membrane penetration capability and subcellular organelle positioning accuracy, and is easy to be distributed in cytoplasm in a nonspecific manner, secondly, acidic environment-induced molecular protonation can significantly change the electronic structure of the probe, so that absorption/emission spectrum deviation and fluorescence quantum efficiency are reduced, thirdly, complicated redox crosstalk exists between HSO 3- and other biological thiols (such as glutathione and cysteine) and active sulfur species (such as H 2S、SO32⁻), and the conventional probe is difficult to realize specific identification. In recent years, functional materials with AIE characteristics provide a new path for breaking through the bottleneck because of the unique advantage of 'hydrophobic aggregation to enhance luminescence'. Compared with an ACQ type probe with a planar conjugated structure, the AIE probe can maintain high brightness in a lysosome acidic water-rich environment through an intramolecular motion limited effect, and meanwhile, accurate positioning of subcellular organelles is realized by virtue of modification of a targeting group. More importantly, through rationally designing the molecular rotor structure and the response site of the probe, the fluorescence signal amplification of the HSO 3- specific activation can be realized, and multidimensional microenvironment parameters such as lysosome viscosity, pH and the like can be synchronously perceived. The research in the field still has obvious technical limitations in that the synthesis strategy of the existing probe depends on complex multi-step modification, and the cooperative optimization of a lysosome targeting module, a signal response unit and an AIE luminescent core is difficult to consider. In addition, conventional detection systems often require exogenous stimuli (e.g., hydrogen peroxide pretreatment or ionophore intervention) to enhance probe response sensitivity, and such manipulations are extremely prone to damage to lysosome membrane integrity, resulting in imbalance in ion homeostasis, and ultimately in distortion of detection signals. Therefore, the development of AIE probes which can adapt to the acidic environment of lysosomes, have in-situ activation characteristics and do not need external intervention can be a key break through the barrier of the prior art and realize the accurate analysis of the metabolic network of the lysosomes HSO 3-. . Disclosure of Invention Aiming at the defects and shortcomings of the prior art, the invention provides a preparation method and application of an AIE fluorescent probe for detecting HSO 3- by targeting lysosomes, wherein weak alkalinity of morpholine ring groups in the probe and weak acidity in cell lysosomes are mutually attracted to realize lysosome targeting, the fluorescent probe can realize specific detection of HSO 3- by modifying a molecular structure, so that the fluorescent probe has high selectivity, and meanwhile, fluorescent groups in the fluorescent probe are blue-shifted from red light to blue light before and after detecting HSO 3-, so that effective observation can be realized through color change during fl