Development of fluorescent chemosensors based on different signal transduction mechanisms

  • Qihua You

Student thesis: Doctoral Thesis


A series of fluorescent probes based on different signal transduction mechanisms for the detection of Fe3+, Zn2+, histidine and pH was designed and synthesized. Their photophysical properties, binding abilities and the further application in cell imaging were fully evaluated.Building on the groundwork of our previous study, molecular scaffold 19 has been appended to spirobenzopyran fluorophore to furnish a highly selective and sensitive Zn2+ sensor. To broaden the application scope of this trifunctional receptive molecule, 19 was incorporated onto rhodamine, antipyrine and coumarin moieties to give 20, 21 and 23, respectively. Probe 20 operative on a chelation-enhanced fluorescence mechanism exhibited highly selective response to Fe3+ with 2:1 stoichiometry of 20-Fe3+ complex. However, a possible tendency of probe 20 to hydrolyze induced by Fe3+ and the unsuccessful attempt of cell imaging would limit its application scope.Probe 21 with O-N-N-N-N-ligand showed a highly selective and sensitive detection of Zn2+. The probe displayed suppressed response to Cd2+ which is the most common interference ion in zinc metal detection. The binding of Zn2+ to probe 21 inhibited the photoinduced electron transfer process originating from the lone pair of the nitrogen atom in the antipyrine moiety to quinoline fluorophore. Therefore, a turn-on fluorescent probe was developed. A moderate binding constant with 1:1 stoichiometry of 21-Zn2+ complex was established by fluorescence titration. The binding mechanism was fully explained by 1H NMR titration. To our delight, probe 21 was successfully applied for recognizing Zn2+ in living cells.The preparation of probe 23 was achieved by appendage of 19 to coumarin derived fluorophore and the probe exhibited a good selectivity and fluorescent turn-off property to Cu2+. The 1:1 stoichiometry of 23-Cu2+ ensemble can serve as an efficient probe for the detection of histidine and biothiols. In the presence of NEM, the influence of biothiols could be eliminated. Furthermore, this sensing ensemble was also used in the detection of histidine in hard-to-transfect U87MG cells with very low cytotoxicity.Based on our group’s previous work on the spiropyran platform, a novel ratiometric near-infrared pH probe 27 operating on an excited-state intramolecular electron transfer mechanism was developed. The pKa was calculated to be 5.9 and the ring-opening/ring-closing mechanism triggered by protons was reasonably explained by 1H NMR titration. However, this spiropyran-based probe was found to be unsuitable for cell imaging.To continue the innovation of pH sensing and extend its application in bioimaging, a series of ratiometric pH probes 32 and 38 characterized by their high quantum yield working in the NIR range was developed. The appendage of N,O-disubstituted hemiaminal ether moiety onto coumarin fluorophore with C=C double bond conferred the sensory material with the ability to display a pH-dependent ratiometric output operating on the ring-opening/ring-closing mechanism. The pKa of 32 and 38 were 6.9 and 5.8 – 6.0, respectively, which rendered them suitable for pH measurement in near-neutral and acidic media. A preliminary work of intracellular pH measurement was also conducted and promising results were obtained
Date of Award9 Jul 2014
Original languageEnglish
SupervisorWing Hong CHAN (Supervisor)

User-Defined Keywords

  • Chemical detectors
  • Design and construction
  • Fluorescent probes

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