Synthesis Of 9-Morpholine-Pyronin And Its Reversible Fluorescence Response To Hypochlorous Acid/Glutathione

In recent years,fluorescent probes have been widely used in the field of chemical biology by virtue of their own perfect temporal-spatial resolution,which can achieve micro-nondestructive detection and other advantages.Pyronin is a commonly used fluorescent dye in biological labeling and imaging for its good light stability and high fluorescence quantum yield.The quaternary ammonium salt structure included in the structure has been certified as a group with mitochondrial positioning function.A series of fluorescent probes are obtained by modifying the 9th position of pyronin Pyronin-9-thione is the key synthetic intermediates for the development of various fluorescent probes for biological labeling and imaging.In this paper,metal sulfides(potassium sulfide,sodium sulfide)are used as inorganic sulfur sources to react in the N,N-dimethylformamide system to prepare Pyronin-9-thione.The reaction time is short and the yield could reach as high as 90%.Further research has shown that S3^·-is the key species for the construction of carbon-sulfur double bonds in the reaction,then the reaction mechanism is inferred based on experimental phenomena and theoretical calculations.Compared with the reported methods,this method greatly simplifies the synthesis steps,reduces the reaction toxicity,and improves the reaction yield.Reactive oxygen species(ROS)are important signaling molecules in cells,including O2^-·,^·OH,¹O₂,H₂O₂,HOCl/Cl O^-,ONOO^-,etc.,which play an important role in maintaining the normal physiological functions of the body and show abnormal expression in pathological processes such as inflammation and inflammation.Excessive production of ROS can cause oxidative stress and destroy cell homeostasis.Glutathione is located in living cells and plays an important role in maintaining the dynamic redox environment of cells and repairing or preventing oxidative damage that leads to cell death or dysfunction.Endogenous glutathione protects cells from damage caused by HOCl.However,the biological activities of HOCl and GSH have not been fully established.In this thesis,9-morpholine-pyronine(abbreviation:MP)was synthesized by modifying the 9-position of pyronine.MP itself has yellow fluorescence(?_ex=492 nm,?_em=560 nm).When it is oxidized by HOCl,blue light was gained(?_ex=392 nm,?_em=455 nm),if reduced by GSH,it emits red light(?_em=605 nm).The probe is a reversible fluorescent probe that can detect HOCl and GSH in cells.In addition,MP has low detection limit,high sensitivity,and has good mitochondrial and lysosome localization function,which has certain significance for the quantitative detection of intracellular redox processes.