| Organic small-molecule fluorescent probe has the characteristics of excellent sensitivity,good selectivity,short response time and transient in-situ non-destructive detection,and are widely used in the detection and analysis of substances in various fields.Including quantitative monitoring of metal ions,small molecules and additives in the field of food safety,gas emission in the field of environmental sanitation,treatment and monitoring of waste water and waste,and analysis of genetic material and organic metabolism in organisms.Organic small-molecule fluorescent probes are usually used in combination with fluorescence imaging technology,and occupy a certain position in the field of bioimaging due to their high biocompatibility and strong tissue penetration.As a biomarker related to various diseases,the development and performance improvement of enzyme detection methods have become a major focus in the field of scientific research in the world today.Therefore,the design and development of more fluorescent probes for enzyme activity detection is of great significance for disease diagnosis and treatment.In this paper,referring to the reported design ideas of various fluorescent probes for enzyme activity detection and their application in bioimaging,new sensitive fluorescent probes corresponding to different enzymes were prepared by using the catalytic reaction mechanism of enzymes.The synthesis steps are simple,the cost is low,and it has excellent performance in cell imaging and enzyme-related disease model imaging.The content is mainly divided into the following three parts:1.The new highly selective red-emitting fluorescent probe(DCDHF-Ala)for Leucine aminopeptidase(LAP)has been synthesized based on 2-dicyanomethyldiene-3-cyano-2,5-dihydrofuran(DCDHF)as fluorophore,and alanine(Ala)as the detection group,has a low detection limit(0.20 U/L),excellent water solubility and cell membrane permeability.In addition,the refined probe has been successfully applied to fluorescent imaging of living cells and zebrafish.DCDHF-Ala has a high biosafety and enables a real-time detection of LAP levels in mice model.2.Developed two novel bioassay-based fluorescent probes for monitoring BCh E in vitro and vivo in an enhanced model.The refined probes DCD-P1 and DCD-P2 used cyclopropanecarboxylic acid chloride as detection group,not only possessed excellent discrimination between BCh E and other analytes but also had high specificity,excellent cell membrane permeability and very short response time(7 min)for BCh E.Moreover,DCD-P1 has been used to image BCh E in living cells,brain tissues and mouse models successfully.3.Developed a novel activatable near-infrared(NIR)fluorescent probe(Chy-1)for specific detection BCh E activity.Excellent sensitivity,good biocompatibility and lower limit of detection(LOD)of 0.12 ng/m L made the probe illustrated extremely specific for the BCh E and successfully used to biological imaging.What's more,Chy-1 can not only clearly distinguish tumor cells from normal cells,but also form a clear boundary between normal tissues and cancer tissues due to the obvious difference in fluorescence intensity produced via in situ spraying.Most important of all,Chy-1 was also successfully applied to track the BCh E activity in AD mouse model.Due to the wide application of organic small-molecule fluorescent probes in the detection of enzyme activitiy,these novel probes studied in this paper may become powerful tools for clinical diagnosis and treatment of tumors and neurodegenerative diseases. |
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