Synthesis And Photo-physical Properties Of Novel Enzyme-reactive Fluorescent Probes

It is well known that various chemical and biological activity of the enzyme are very important diagnostic indicators in clinical testing. The highly sensitivity, visibility, simple operation of the fluorescent probe are more and more being concerned with the technical progress of fluorescent probe. Especially in recent years, fluorescent probe technology are more and more extensively being applied to clinical medicine, biology and other fields, used in cell biology, bio-medical as specialty drugs and diagnostic reagents.2-(2-Hydroxyphenyl) benzimidazole (HPBI) and its derivatives are a series of important fluorescent compounds in the case of their intense emission properties via excited-state intramolecular proton transfer (ESIPT). Meanwhile, the HPBI derivatives are useful as a new series of fluorescent probes, high-energy radiation detector, plastic scintillators and polymer ultraviolet stabilizers. So it is of great importance to study the synthesis and fluorescent characteristics of these compounds.This artical includes two parts as follows:1. HPBI derivatives (2a~2d) were synthesized with sub-salicylaldehyde and 1,2-phenylenediamine as the raw materials. (2a~2d) entered into reaction with phosphorus oxychloride in pyridine prior to hydrolysis in an alkaline medium to yield the targer compound—2-benzimidazolephenylphosphate and better than the AMP-buffer method used for the determination of alkaline phosphatase activity to raise the accuracy of analysis.2. A convenient method to synthesize three kinds of novel 2-(2-hydroxyphenyl) benzimidazole in order to improve the solubilities of the fluorophores with substituent(s) of carboxyl, ester and double-bond which are aimed for determining the sulphydryl group was reported. The corresponding photo-physical properties were characterized by ultraviolet absorption and fluorescence spectra in methanol. They also be used for–SH in HepG2 and L-Cys, and the cellular experiment results showed that BIHPMA, BIHPPA and BIHPM can be used as fluorescent probes for–SH in cells.