The Design And Synthesis Of Novel Calcium Ion Fluorescence Probes

Calcium is a divalent metal ion, which is abundant in human cells. Ionic calcium (Ca2+) act as important messenger in cells. Calcium ions involved in a number of physiological processes, such as cell division and differentiation, metabolism, etc. Therefore, it is essential to accurately determine the concentration and spatial distribution of the intracellular calcium. This laid foundation of many studies of diseases, physiological and pathological processes.To date, the most common method for determining intracellular calcium is based on small molecule fluorescent probes. In particular, the most famous calcium fluorescent indicators are the second (Fura-2) and third generation (Fluo-3) ones developed by Tisen and coworkers. However, the excitation wavelength of Fura-2is in ultraviolet region, which can damage the cells. As for Fluo-3, it detects ionic calcium in the mode of fluorescence enhancement, which cannot provide quantitative information of ionic calcium concentration. Currently, there is few reports of excellent performance calcium ion fluorescent indicators. Therefore, novel calcium ion fluorescence probes need to be designed and synthesized.In this thesis, we have carried out the following researches:(1)O-nitro phenol was used as the starting material, we have synthesized the important intermediates BAPTA-CH3-CHO and BAPTA-CH3-NH2. Two intermediates were adequately characterized by1H NMR,13C NMR and MALDI-TOF-MS. Meanwhile, we have studied the reaction conditions of the synthesis of Two intermediates, have improved synthesis methods, have optimized the reaction conditions, and have increased the reaction yield.(2)BAPTA-CH3-CHO, as the receptor of Ca2+, was introduced into BODIPY fluorescent chromophore to develop Ca2+fluorescent probes BODIPPY-S and BODIPY-D. Two probes were adequately characterized by1H NMR,13C NMR and MALDI-TOF-MS. Their maximum emission wavelength were646nm and709nm respectively.(3)BAPTA-CH3-NH2, as the receptor of Ca2+, was introduced into perylene fluorescent chromophore to develop another Ca2+fluorescent probe PDI-Ca. The structure was adequately characterized by1H NMR,13C NMR and MALDI-TOF MS.