基于检测碱性磷酸酶的荧光探针的合成及其生物评价

Synthesis and Biological Evaluation of Fluorescent Probes Based on Detection ofAlkaline Phosphatase

目的:本实验利用磷酸酯为碱性磷酸酶(Alkaline Phosphatase,ALP)的反应位点,DCM类染料为荧光发色团,构建能特异性检测ALP的荧光诊断试剂。方法:以2-甲基-6-叔丁基-4H-吡喃酮为起始原料,经过缩合、取代反应得到目标化合物探针X。采用紫外光谱法与荧光光谱法,在化学体系中对其吸收、发射波长进行测定;MTT实验验证其细胞毒性,细胞成像实验检测活细胞中的碱性磷酸酶。结果:所得产物经核磁共振氢谱和碳谱、高分辨质谱鉴定了结构。细胞成像实验结果表明探针X可用于检测活细胞中的碱性磷酸酶。

Objective: In this experiment, phosphate was used as the reaction site of Alkaline Phosphatase (ALP), and DCM dyes were used as fluorescent chromophores to construct a fluorescent diagnostic reagent that can specifically detect ALP. Methods: Using 2-methyl-6-tert-butyl-4H-pyrone as the starting material, the target compound probe X was obtained through condensation and substitution reactions. Ultraviolet spectroscopy and fluorescence spectroscopy were used to determine its absorption and emission wavelengths in a chemical system;MTT experiments were used to verify its cytotoxicity, and cell imaging experiments were used to detect alkaline phosphatase in living cells. Results: The structures of the obtained products were identified by high resolution mass spectrometry, nuclear magnetic resonance proton spectroscopy and carbon spectroscopy. Cell imaging experiments show that probe X can be used to detect alkaline phosphatase in living cells.