Design,Synthesis And Application Of Cu²⁺,PSA And Thiophenol Fluorescence Probes

BackgroundCopper ions widely exist in nature and life,and their existence plays an important role in the sustainable development of nature and the maintenance of life health.Thiophenol is used as an intermediate in fine chemicals such as dyes,pharmaceuticals,pesticides,polymerization inhibitors,antioxidants,etc.,and has attracted attention due to its extremely toxic properties.Prostate specific antigen(PSA)is the first choice for the diagnosis of prostate cancer because of its high tissue and organ specificity and species specificity.It is also used for forensic seminal plaque identification.Fluorescence detection is widely used in many fields such as life science,medicine,environmental science and so on,because of its high selectivity,low detection limit,low toxicity and side effects,as well as the characteristics of in-situ real-time visualization and intelligent analysis.It has been widely used in the identification and detection of ions,molecules and even biological macromolecules in the fields of life science,medicine,environmental science and so forth.Therefore,the design and synthesis of fluorescent probes for copper ion,thiophenol and prostate specific antigen have become the focus and hot spot in the research of organic small molecular probes.ObjectivesDesign and synthesize high-selectivity,low detection limit and specific visualization of copper ion,thiophenol and a series of prostate-specific antigen fluorescent probes,and apply cell imaging experiments and semen sample imaging experiments to the practical application value of the corresponding probes.Methods1.Design and synthesis of copper ion fluorescent probes L1,L2.Design and synthesis of thiophenol fluorescent probe L3.Design and synthesis of a series of prostate specific antigen fluorescent probes 1-7.The probes were structurally characterized by mass spectrometry,nuclear magnetic resonance,high performance liquid chromatography,and the like.2.Ultraviolet-visible spectroscopy and fluorescence spectroscopy were used to measure the response of the corresponding probes to copper ions,thiophenols and prostate specific antigens.3.The HepG-2 cells and SiHa cells were cultured,and the semen samples with different attachments were prepared.The biomedical application value of the probes was investigated by using an inverted fluorescence microscope.Results1.Copper ion fluorescence probe:Two copper ion fluorescence probes were successfully designed and synthesized.The fluorescence probe L1(a compound based on coumaryl hydrazide and 3-aldehyde indazole)could recognize copper ions by fluorescence quenching,but there’s interference with mercury.The fluorescence probe L2(a dual-arm compound based on biphenyl dihydrazone and 9-formyl-8-hydroxyduronidine)also could recognize and detect copper ions by fluorescence quenching.However,because the two-arm structure of L2 was more sensitive and strong in binding to copper ion,the probe had better selectivity and lower detection limit The probe L1 and L2 were co-incubated with cells and observed under the inverted fluorescence microscope.The results showed that both probes could be used for the quantitative measurement of copper ions in the biological system.2.Thiophenol fluorescent probe:A reactive thiophenol fluorescent probe L3 was successfully designed and synthesized.The probe had obvious aggregation-induced luminescence(AIE)phenomenon and intramolecular proton transfer(ESIPT)effect in the process of recognition and detection of thiophenol.The fluorescence emission wavelength and large stokes shift in the near infrared region were also observed.There was a good linear relationship between the fluorescence intensity of the probe and the concentration of thiophenol(1.0 x 10.0 μM).The minimum detection limit was 4.5 × 10-7 M,and the probe could be considered as a sensitive detector for thiophenol.3.Prostate specific antigen fluorescent probe:Based on the mechanism of fluorescence resonance energy transfer(FRET),a series of fluorescent probes for prostate specific antigen 1-7 were synthesized.Two targeted fluorescent probes,4 and 6,with good response to prostate specific antigen were screened by fluorescence spectrum test.The response time of probe 4 is faster and the sensitivity is higher.The practical application value of probe 4 was verified by the application of probe 4 in semen sample.Conclusions1.Two copper ion fluorescent probes were successfully designed,synthesized and applied to cell fluorescence imaging experiments;2.A sample of thiophenol fluorescent probe was successfully designed and synthesized,which was expected to be used for the detection of water pollutants;3.A series of prostate specific antigen fluorescent probes were designed and synthesized.Two of the sensitive fluorescent probes were screened,laying the foundation for optimization and improvement and development of near-infrared multifunctional fluorescent probes.