The Design,Synthesis Of Reversible Fluorescent Probe And Application In Drug Research

Intracellular glutathione(GSH),the most abundant nonprotein thiol(1-10 mM)in cells,plays pleiotropic roles in maintaining intracellular redox homeostasis through equilibrium between its reduced thiol form(GSH)and its oxidized disulfide form(GSSG).Hnece,real time detection of variations in cellular levels of GSH is good for understand changes of redox state inside cells.Until now,most existing fluorescent probes can be utilized to determine GSH in only a qualitative manner,unsuitable for real time quantification of variations in the levels of GSH.To overcome this issue,we design and synthesize a new type of reversible fluorescence probe based on a,?-unsaturated ketone compound,and researches the changes of GSH content in vitro and in cells which achieve the analysis of GSH in situ,real time and dynamic fluorescence imaging.Through the regulation of structure,the reactivity of probe is controlled.One of them,the cyanoacrylamide compound 4d can react with GSH in reversible addition manner with the dissociation constant at 2.59 mM.By using this probe,the concentration of GSH in Hela cell is caculated(5.40 ± 0.87 mM)and tracked.In addition,the cyanoacrylate compound 4c which is a reproducible fluorescent probe with high utilization rate can recognize the bisulfite specifically.Using this reversible fluorescence detection of bisulfite,it is facilitate to track the status of bisulfite-induced disease and process of treatment which bring a great help to understand the causes of the disease and the effect of treatment.Couple the oleanolic acid with reversible GSH fluorescent probe to understand on the intrinsic relationship between the drug action and GSH.From the fluorescence imaging study,it could reveal the real time viration of GSH in the process of drug action which help to explore the relationship between GSH level and efficacy of drug.Bond the oleanolic acid with non-reactive probe to study the specificity of the drug and the changes of oxidative stress in the cell.In a word,this study explored the problem of quantitative and real time detection of GSH by reversible fluorescence probe,and coupled it with the drug to explore the mechanism of drug action by using the anticancer activity of the drug and the localization or identification ability of the probe.