Synthesis Of Active Site-oriented Fluorescent Labeled Molecules Of Matrix Metalloproteinase-7

The degradation of extracellular matrix is essential for the growth,invasion,metastasis and angiogenesis of malignant tumors.Matrix metalloproteinases(MMPs)are a family of zinc-dependent neutral endopeptidases that can jointly degrade almost all matrix components.It can be detected in the tumor tissue of patients with advanced cancer or to elevated serum levels of various MMPs and their role has been widely examined as a prognostic indicator of cancer.Matrix metalloproteinase(MMP-7),which also known as matrix lysin,is the smallest molecular weight enzyme in the matrix metalloproteinase family,and exhibits proteolytic activity on the components of the extracellular matrix(ECM).A tumor is related to cell growth,invasion,metastasis and angiogenesis multi-step process.In this process,matrix metalloproteinase-7 is usually overexpressed in tumor cells.By degrading elastin in blood vessel walls and lymphatic vessel walls,it plays an important role in tumor invasion and metastasis,and is closely related to cancer progression.Activity-based protein profiling(ABPP)is a powerful post-genomic technology that uses active site-specific chemical probes to monitor enzyme functions in complex biological systems.The probe for specific fluorescent labeling of MMP-7 was designed based on the ABPP method.It can study cell processes and molecular dynamics in real time at the single cell level,and also produce a more comprehensive understanding of the cell environment where protein levels in tumor tissues have changed at the tissue level.Based on the existing literature reports and the previous results of this research group,this paper designed and synthesized a new type of active site-oriented fluorescent labeling molecule of matrix MMP-7(compound 13).The molecule consists of six small molecules: L-dopa,piperazine,4-fluorobenzenesulfonyl chloride,diethyl isobutylmalonate,N,N-dimethyl meta-hydroxyphenol,and L-tert-leucine.It synthesized through 20-step reactions such as condensation,esterification,substitution,and reduction.Based on the ABPP method,pyronin is used as the fluorescent reporter group,and diethyl isobutylmalonate,L-tert-leucine and L-dopa are used as reactive groups to design and synthesize a new fluorescent probe for real-time detection MMP-7.It is expected that compound 13 can selectively label MMP-7 and determine the temporal and spatial distribution of MMP-7 in a specific cancer pathology model,thereby revealing the mechanism of occurrence and development in the cancer pathology model.Thus providing strong theoretical support for the development of new in vitro diagnostic reagents.