Design,Synthesis And Activity Assay Of The Bioactive Fluorescent Probes Based On The TPE

Activated protein is the basic ingredient of life,whose specific detection is an important topic in chemical biology.Because of their high sensitivity,good selectivity,non-invasive detection and the requirement of a tiny amount of sample,fluorescent probes have been attracted wide interests.Compared with traditional fluorescent molecules which have an aggregation-caused quenching(ACQ)effect,the new fluorescent molecules tetraphenylethene(TPE)could improve the sensitivity with aggregation-induced emission(AIE)effect.In this thesis,a series of fluorescent probes based on TPE were successfully designed and synthesized,and their applications in the detection of proteins,such as protease,monoamine oxidase,phosphatases were further studied.This thesis includes the following parts:1.Methoxypolyethylene glycols(mPEG,1000)was used to increase the water solubility of probes to reduce the fluorescence signal,then based on the research method that fluorescence enhanced when the probe was combined with protein,five different TPE probes were synthesized with different charge amino acids.The results show that probe 1-5 were combined with protein to enhance fluorescence intensity,therefore,the probes sensitive to isoelectric point.2.Three water-soluble TPE probes for monoamine oxidase(MAO)were designed and synthesized.The 4-methyl pyridinium was used as monoamine oxidase affinity group and TPE was linked by three different spacers.The results revealed that these three probes were able to test the activity of monoamine oxidase.Furthermore,the probe with phenyl spacer and trans structure displayed the highest selectivity to MAO-A.Cellular imaging studies also proved that the probes was able to monitor the activity of MAOs in living cells.3.In terms of the principle of fluorescence resonance energy transfer(FRET),we designed and synthesized a "OFF-ON" FRET probe that phosphate group was regarded as the substrate of alkaline phosphatase,TPEs and 5(6)-carboxyfluorescein were as donor and acceptor respectively.The results displayed that the two fluorophores were a good pair of FRET and the probe showed the highly specificity to alkaline' phosphatase.Probe was able to report alkaline phosphatase activity in living cells directly.In conclusion,we successfully synthesed nine probes,probe 1-5 could be used to measure the positive and negative protein;probes6,probe7a and probe7b were able to test the activity of MAO-A;probe8 had the specificity detection of alkaline phosphatase.