The Construction Of Novel Peroxynitrite Fluorescent Probes And Thier Application In The Detection Of Biomarks

In recent years,fluorescent probes have been widely used in chemical medicine,environmental science,disease diagnosis and bioimaging,especially in real-time detection of samples and fluorescence imaging of biological cells,due to their unparalleled advantages of fast analysis speed,high spatial and temporal resolution,good biocompatibility and non-destructive in situ imaging.In this paper,we designed and synthesized three new types of fluorescent probes to detect the ONOO^-and successfully applied to the imaging of actual biological samples.1.We first designed and synthesized a novel ratiometric fluorescent probe（Cy-NH₂）for the detection of ONOO^-.The probe has a simple structure and can be obtained by a simple procedure;the destruction of theπ-conjugated system due to the strong oxidizing ability of ONOO^-causes the probe to have a large emission wavelength shift（248 nm）;in addition,the probe has high sensitivity with ONOO^-reaction,and the fluorescence intensity ratio is as high as 1728-fold;more importantly,the probe has been successfully applied to the fluorescence detection of exogenous and endogenous ONOO^-in living cells,and satisfactory results have been obtained.2.Based on the excellent performance of the probe in our previous chapter,we improved the synthesis method and synthesized a series of ratiometric fluorescent probes with different substituents,from which the best fluorescent probes（Cy-NEt₂）were selected.First of all,our probes can be synthesized de novo by reliable and inexpensive raw materials.In addition,the probe exhibits a large emission wavelength shift（255 nm）,which is beneficial for enhancing the signal background ratio.Furthermore,Cy-NEt₂ can specifically target mitochondria and thus can be used to assess mitochondrial oxidative stress in cells and mice.3.We designed and synthesized a new near-infrared fluorescent probe（DCP-B）for the detection of ONOO^-.Compared to the probes from the previous two chapters,the probe has larger Stokes shift（173 nm）,longer near infrared wavelength（763 nm）and lower detection limit（3.3 nM）.In the probe,we chose the excellent dicyanoisophorone derivative（DCP-OH）as the fluorophore and the borate group as the recognition unit.The probe DCP-B itself has no fluorescence because the intramolecular charge transfer（ICT）process is interrupted.When ONOO^-is added,the borate reacts with ONOO^-,and the ICT process recovers,thereby emitting strong near-infrared fluorescence.Due to the excellent performance of the probe,we successfully applied it imaging to exogenous and endogenous ONOO^-in living cells and achieved satisfactory results.