Synthesis And Interaction Of Enzyme With Perylene Monomimide-Sugar Compounds

Cancer is one of the most serious health problems in the world.In recent years,the incidence of malignant tumors in China has increased by 3.9%annually and the mortality by2.5%per year.After normal cell produces cancerous change,the normal physiology environment will change.Abnormal enzyme metabolism,partial acidity and hypoxia are all characteristics of tumor microenvironment.Therefore,early diagnosis is very important for the treatment of tumor patients.Fluorescence-based optical probe can be used to real-time monitor the target enzymes in the body of a biological process.Therefore,in recent years,many related fluorescent probes have been synthesized against nitroreductase and glutamate transpeptidase,but these probes are only used to monitor,and there are few probes for monitoring that can both monitor and treat,Therefore,the research space of enzyme-induced therapy is very large.Perylene monoimides derivatives can be used as probes as well as for treatment.Meanwhile,perylene monoamide?PMI?has good photostability and multiple modification sites,which can be connected with different groups to regulate their absorption wavelength to reach near-infrared.In order to accurately target tumor cells to achieve selective diagnosis and treatment,sugar chain branches are selected.Modification of perylene monoimide with mannose or lactose can also increase the water solubility and biocompatibility of perylene derivatives.Based on this,several compounds were designed and synthesized from nitroreductase and?-glutamine transpeptidase,and their enzymatic reactions in vitro were studied.The specific contents are as follows:1.Nitro-PMI-sugar conjugates?PMI-1NO₂-Man,PMI-1NO₂-Lac,PMI-2NO₂-Man,PMI-2NO₂-Lac?were synthesized through a multistep reactions such as esterification,nitration,hydrolysis,and Click.The imides of perylene monoimides were modified with mannose and lactose,and the 9,10-positions of perylene monoimides were modified with nitro.The optimal conditions for the nitration reaction of perylene monoimide were explored.The compounds were characterized by ¹H NMR,¹³C NMR and HRMS.2.Glutamine-PMI-lactose compounds?P-7-7?were synthesized with several steps such as substitution,hydrolysis,condensation,and Click.Perylene monoimide bays were modified with lactose,and 9-position of perylene monoimide was first modified with3R-aminopyrrolidine,the amino group of pyrrolidine was condensed with glutamate.The compounds were characterized by ¹H NMR,¹³C NMR and HRMS.3.The spectroscopic properties of Nitro-PMI-sugar compounds were studied for ther spectral properties.It was found that the ultraviolet absorption wavelength shifted from480nm to 540nm after the reaction of PMI-1NO₂-Man,PMI-1NO₂-Lac,with nitroreductase.The properties of PMI-2NO₂-Man compounds in aqueous solution cannot be measured due to poor water solubility.After the reaction of PMI-2NO₂-Lac with nitroreductase,the UV absorption was found in all wavelengths,and the absorption was obvious at 800-1050 nm.The effects of different concentrations of NADH and NTR on the enzymatic reaction were investigated;The spectral variation of the reaction time between the compound and the enzyme was also investigated.These compounds,which have almost no fluorescence,can convert absorbed light energy into heat energy for the possibility of photothermal therapy.