Synthesis Of Novel Fluorescent Probe Based On Quinoline Skeleton And Its Application In Environmental Analysis

Fluorescence imaging analysis is an important method to identify and detect biologically active molecules,and has been widely used in the fields of biology,chemistry,environment and materials.Fluorescent probe based on 4-trifluoromethyl-7-aminoquinoline structure exhibits excellent selectivity,stability and low cytotoxicity.At the same time,this kind of probe has targeting ability,which is of great significance for monitoring changes in the intracellular environment and preventing diseases.The main work of this thesis is to design and synthesize several novel Golgi-targeted fluorescent probes based on 4-trifluoromethyl-7-aminoquinoline skeleton,which can be used to detect the specific active molecules,qualitatively and quantitatively in organisms,and be practically applied in biological imaging,etc.The details are as follows:1.By using 4-trifluoromethyl-7-aminoquinoline dye as the fluorophore and Golgi targeting unit,and 2,4-dinitrobenzenesulfonate as the cysteine(Cys)recognition group,we synthesized a kind of Golgi-targeted Cys fluorescent probe Gol-1,and the related recognizing performance for Cys of the probe Gol-1 was studied in detail.The results showed that the probe Gol-1 had good selectivity and stability to Cys in HEPES buffer(10 mM.pH=7.4),and exhibited obvious fluorescence "off-on" changes after interacting with Cys.In addition,the probe Gol-1 has satisfactory Golgi apparatus targeting ability,and exhibits low toxicity and good permeability in cell imaging,which can be used to monitor Cys levels during Golgi stress in living cells.2.The azo group can undergo cleavage reaction under the catalysis of azoreductase,and can be used as the responsive group of azoreductase in the construction of fluorescent probes.By using 4-trifluoromethyl-7-aminoquinoline dye as the parent group,the fluorescent probe Gol-2 was synthesized by linking N,N-dimethyl-p-phenylenediamine through azo bonds.In the hypoxic environment of cells,the probe Gol-2 exhibited excellent selectivity for azoreductase in the EtOH:HEPES(5:5,pH=7.4,10 mM HEPES)system.Upon the addition of azoreductase,the azo group of the probe was converted to amine group through the reduction reaction,which can reverse the fluorescence of the probe.The probe can be applied as an ideal candidate for detecting azoreductase in enviroment and biological processes.3.Taking advantage of the property of quenching fluorescence by photoisomerization of azo groups,using 4-trifluoromethyl-7-aminoquinoline dye as the backbone,N,N-bis(dichloroethyl)-1,4-benzene Diamine is a pharmacophore,and a fluorescent probe Gol-3 for the detection of azoreductase and targeted controlled release was synthesized by chemical coupling of azo bonds.By simulating the physiological environment in hypoxic tumors,the fluorescence spectrum of the probe Gol-3 was recorded with DMSO:Tris(10 mM,pH=7.4,v/v=5:5)as the test system,and the results showed that the probe Gol-3 have specific selectivity for azoreductase.Similar to the mechanism of probe Gol-2,under the action of azo reductase,the azo bond is broken,releasing fluorophore and DNA alkylating active drug.Theoretically,it is one of the few fluorescent probes that has a certain therapeutic effect on tumors.