Synthesis And Study Of Fluoroboron Fluorescent Organic Small Molecular Probes

In recent years,the identification of important analytes in the environment and organisms and their mechanism of action has been an important research topic.Compared with other analytical tools,fluorescence detection technology has several advantages,including simple application,sensitive detection,and most importantly real-time dynamic cell imaging with confocal microscopy.The main purpose of this thesis is to use BODIPY as the fluorophore and to select the appropriate linker and recognition group so as to obtain a metal ion recognition probe and an active oxygen recognition probe with good fluorescence performance,high specificity,recognition in aqueous solution and apply it to biological imaging.In order to study the metal ions and reactive oxygen species in vivo metabolism,biological function and toxicology provide a powerful tool.In this dissertation,BODIPY fluorescent dyes were used as fluorophores,macrocyclic polyamines as recognition groups,and connection by appropriate connecting arms to obtain water-soluble probes with specific recognition of silver ions.The probe has excellent selectivity and sensitivity to silver ions,and has low cytotoxicity and can be used for imaging silver ions in cells,which is of great help in revealing the mechanism of action of silver ions in vivo.In the synthesis of the probe BOIDPY-Ag,the by-product in the hydrogenation process was structurally characterized to obtain the ring-opening addition product of tetrahydrofuran,and the fluorescence property was measured to obtain the copper ion recognition probe.The probe has good selectivity and is sensitive to copper ions with a detection limit of 25 nm.And this probe has low cytotoxicityn.So it can be used for intracellular copper ion imaging.During the selection of the probe BOIDPY-Ag arm it was found that when the amide group on the benzene ring of the BOIDPY group forms an amide bond.The obtained material has a very strong fluorescence signal and has excellent fluorescence performance.Therefore,we chose this material as a fluorescent chromophore,and connected pyridinium ions and triphenylphosphine cations as mitochondria targeting groups to synthesize mitochondrial targeting probes.Experiments show that the probe BODIPY-Py fluorescence quantum yield in aqueous solution,fluorescence signal stability,and low cytotoxicity of the probe can be applied to cell imaging.Thus,the probe can serve as a mitochondria-targeting fluorescent probe and verify the mitochondrial targeting of pyridinium ions.Based on the mitochondrial targeting of pyridinium ions,fluoroboropyridine derivatives were used as fluorophores and phenylboronic acid as recognition groups.The two groups were connecting by the pyridinium salt linkage structure to designed and synthesized BODIPY-BB.The mechanism of recognition was identified by high-resolution mass spectrometry.The probe showed good water solubility and an "off-on" response mechanism that allowed the selective recognition of CkO-without interference from other reactive oxygen species reactive nitrogen molecules.There was an excellent linear relationship between fluorescence signal intensity and ClO-concentration.The detection limit is 0.6 pM.In addition,the recognition is rapid and the fluorescence signal is stable.Cell experiments show that BODIPY-BB cytotoxic low,and has excellent mitochondrial targeting,so it can be applied to vivo ClO-imaging.Subsequently,based on the protection mechanism of the thiocarbamoyl-p-phenol group,the recognition group was replaced by dimethylamino-p-thiocarbonylbenzyl alcohol.The probe BODIPY-DMTC was designed and synthesized.By HNMR and HRMS,we verifed the probe's protection-deprotection recognition mechanism.The probe showed good water solubility and an "off-on" response mechanism that allowed the one-way recognition of ClO-without interference from other reactive oxygen species reactive nitrogen molecules and an excellent linear relationship between fluorescence signal intensity and ClO-concentration.The detection limit is 95 nM,and recognition is rapid and the fluorescence signal is stable.Cell experiments show that BODIPY-DMTC low cytotoxicity,has excellent mitochondrial targeting.And the probe is sensitive and can be applied to the detection of endogenous ClO-and biological living body imaging.Therefore,this probe can identify the physiological and pathological processes and functions of ClO-at organelle level or at the level of biological tissues.Peroxynitrite is a high-energy reactive oxygen species that is involved in the pathogenesis of many diseases.In this thesis,a near-infrared fluorescent probe(DCM-PN)was designed and synthesized using dicyanomethylene-4H-pyrans as a fluorescent group to detect the concentration of endogenous peroxynitrite in vivo.The probe has an extremely fast response rate(5 S),extremely high sensitivity(53 nM)and excellent specificity,and the probe can be successfully used for cell imaging and mouse model imaging.Therefore,the probe can be an effective tool for studying the physiological and pathological effects of peroxynitrite in vivo.