CN-122010923-A - Benzothiadiazole fluorescent probe and preparation and application thereof

CN122010923ACN 122010923 ACN122010923 ACN 122010923ACN-122010923-A

Abstract

The invention discloses a benzothiadiazole fluorescent probe and preparation and application thereof. According to the invention, through a two-step Suzuki coupling reaction, 4-pyridyl and N-methylpiperazine groups are sequentially introduced, and the benzothiadiazole fluorescent probe for lysosome polarity detection is simply, conveniently and efficiently synthesized. The probe has obvious positive solvent color effect, and the fluorescence intensity of the probe is reduced along with the increase of the polarity of a medium, so that the specific fluorescence enhancement can be realized in a low-polarity lysosome microenvironment. The probe has excellent light stability and is suitable for long-time dynamic fluorescence imaging monitoring.

Inventors

ZHANG ZIWEI
Wu Maowang
YANG KE
YANG YANYAN
FENG QIANYI
SU LI
FANG SHIYONG
YANG LI

Assignees

皖南医学院

Dates

Publication Date: 20260512
Application Date: 20260130

Claims (10)

1. The benzothiadiazole fluorescent probe is characterized by having the following structural formula: wherein the X group is a substituted or unsubstituted piperazinyl group, and the Y group is a substituted or unsubstituted pyridinyl group.
2. The benzothiadiazole fluorescent probe according to claim 1, wherein the X group is Wherein, R 1 、R 2 、R 3 、R 4 、R 5 groups are all independently selected from one of H, methyl and ethyl.
3. The benzothiadiazole fluorescent probe according to claim 2, wherein the chemical structural formula is as follows: 。
4. The method for preparing the benzothiadiazole fluorescent probe according to claim 1, comprising the following steps: wherein Z is halogen and R 1 、R 2 、R 3 、R 4 、R 5 groups are each independently selected from H or methyl.
5. The method for preparing the benzothiadiazole fluorescent probe according to claim 4, wherein the phase transfer catalyst comprises at least one of tetrabutylammonium bromide, tetrabutylammonium tribromide, tetramethylammonium bromide, tetrapropylammonium chloride and tetrabutylammonium iodide, and the palladium catalyst comprises at least one of tetrakis (triphenylphosphine) palladium, bis (triphenylphosphine) palladium dichloride, tris (dibenzylideneacetone) dipalladium and bis (acetonitrile) palladium dichloride; the R 1 groups are methyl groups, the R 2 、R 3 、R 4 、R 5 groups are H, and Z is Br or Cl.
6. The method for preparing a benzothiadiazole fluorescent probe according to claim 4, wherein the molar ratio of the compound I to the compound II is 0.5-1.0:1.0-2.0.
7. The method for preparing the benzothiadiazole fluorescent probe according to claim 4, wherein the heating reaction condition is that a mixed solution is obtained by mixing a compound I, a compound II, a phase transfer catalyst, a palladium catalyst, an organic solvent and water, and the mixed solution is heated to 80-100 ℃ to stir and react for 12-36 h, and the organic solvent comprises at least one of toluene, xylene, methylene dichloride and dioxane.
8. The method for preparing the benzothiadiazole fluorescent probe according to claim 4, wherein the preparation method of the compound I is as follows: Wherein Z is Br or Cl, and the palladium catalyst comprises at least one of tetra (triphenylphosphine) palladium, bis (triphenylphosphine) palladium dichloride, tris (dibenzylideneacetone) dipalladium and bis (acetonitrile) palladium dichloride.
9. The method for preparing the benzothiadiazole fluorescent probe according to claim 8, wherein the heating condition is that the heating reflux is 12-36 h under the temperature of 80-100 ℃, the molar ratio of the 4-pyridine boric acid to the compound IV is 3-4:3-4, and the solvent for the reaction is a mixture of water and at least one of toluene, xylene, methylene dichloride and dioxane.
10. Use of a benzothiadiazole fluorescent probe according to any of claims 1-3 for targeting lysosomes in a labeled cell and/or for detecting changes in the polarity level of lysosomes in a cell.

Description

Benzothiadiazole fluorescent probe and preparation and application thereof Technical Field The invention relates to an organic small molecule fluorescent probe, in particular to a benzothiadiazole fluorescent probe and preparation and application thereof. Background Polarity is an important parameter for representing the change of the microenvironment of an organism, and is involved in most cellular processes such as catalysis of enzymes, protein activation, lipid composition and the like. Cell polarity is a characteristic feature of the cellular functioning and regulation mechanisms, and when the physiological and pathological activities of the cells change, their polarity changes accordingly. Therefore, detecting cell polarity is of great importance for monitoring different cell states and for facilitating a deep understanding of the relevant physiological and pathological processes. In addition, different organelles within the cell assume specific biological functions, and there are also differences in the polarity levels of their local microenvironments. Therefore, detecting the polarity of different organelles is of great value for accurately resolving changes in cellular state. Lysosomes are important organelles in eukaryotic cells, and the core functions include biological macromolecular degradation, recycling of cellular components, metabolic regulation and other physiological processes. Lysosomal dysfunctions, when present, may trigger pathological conditions such as lysosomal storage disorders, neurodegenerative diseases, cardiovascular diseases, and cancer. As an important degradation organelle in a cell, dynamic regulation of the polar characteristics of lysosome membrane potential, pH value, ion homeostasis, etc. are critical to maintaining cell function. Recent studies have shown that lysosomal polarity abnormalities are closely related to the pathological course of neurodegenerative diseases, cancers, metabolic diseases (e.g. lysosomal storage diseases, diabetes). Therefore, the establishment of a technical method for accurately detecting the dynamic change of the lysosome polarity is of great significance for revealing the regulation and control mechanism of the lysosome polarity in life activities. Fluorescent imaging technology has been widely used in the field of cell biology due to its characteristics of non-invasiveness, high sensitivity, excellent spatial-temporal resolution, and the like. The fluorescent probe is used as a core tool of the technology, and is widely applied to the fields of dynamic monitoring of cell microenvironment, bioactive substance sensing, disease diagnosis, living body imaging and the like by virtue of the advantages of high specificity, real-time visualization, noninvasive detection and the like. In research and application of fluorescent probes, classical fluorescent parent nuclei such as coumarin, BODIPY, rhodamine and fluorescein are often used as core frameworks. As a novel hybridized structure fluorophore, the benzothiadiazole has the photophysical properties of good light stability, high fluorescence brightness, easy structure modification and the like, and has been applied to the design synthesis of fluorescent probes. At present, although related reports about detecting lysosome polarity fluorescent probes exist, the existing researches are mainly focused on obesity and inflammation models, the researches on the polarity level and dynamic change of nerve cell lysosomes are not yet reported, and how to realize quantitative determination of the polarity of nerve cell lysosomes based on small molecular fluorescent probes becomes a technical problem to be solved urgently. Disclosure of Invention The first object of the invention is to provide a benzothiadiazole fluorescent probe which can target lysosomes and is sensitive to dynamic changes of lysosome polarity. The second purpose of the invention is to provide a benzothiadiazole fluorescent probe, which solves the problem of how to prepare the benzothiadiazole fluorescent probe. The third purpose of the invention is to provide application of the benzothiadiazole fluorescent probe in targeting and labeling lysosomes in cells and/or detecting the polarity level change of lysosomes in cells, and solve the problem of how to target and label lysosomes in cells and/or detect the polarity level change of lysosomes in cells. The technical scheme is that the benzothiadiazole fluorescent probe has the following structural formula: wherein the X group is a substituted or unsubstituted piperazinyl group, and the Y group is a substituted or unsubstituted pyridinyl group. Preferably, the X group isWherein, R 1、R2、R3、R4、R5 groups are all independently selected from one of H, methyl and ethyl. Specifically, the chemical structural formula of the benzothiadiazole fluorescent probe is as follows: 。 According to the invention, through a two-step Suzuki coupling reaction, 4-pyridyl and N-methylpiperazine groups are sequentia