近红外比率型氧化还原可逆荧光探针的设计合成及其在细胞内活性氧物种原位动态检测中的应用

Reactive oxygen species play crucial role in the physiological process. However, current research in the relationships between the reactive oxygen species and disease formation mechanism still stays at the macro level. The imaging analysis and quantitative analysis of reactive oxygen species is significant to elucidate the nature of reactive oxygen species and their related chemical biology at the molecular level. Therefore, to develop in situ, real-time, visualization and quantitative analytical methods of reactive oxygen species is a challenge for analytical chemistry. The project proposes a series of near-infrared, ratiometric and reversible redox fluorescent probes with good biological compatibility for reactive oxygen species analysis using the BODIPY as fluorescent units with the tuning molecular structure and superior fluorescence properties. Intracellular reactive oxygen species will be detected by the bioprobe’s fluorescence signal. Thus, combined with the laser confocal imaging technique, a novel, in situ, real-time analytical method for intracellular reactive oxygen species detection will be developed. It is expected to provide one excellent method and technology to understand the relationships between the reactive oxygen species and the diseases under the molecular level.

活性氧物种在生理过程中起着举足轻重的作用，但其与疾病形成机制研究还停留在宏观水平上。活性氧物种的成像分析和定量分析对从分子水平上阐明活性氧物的本质和相关化学生物学研究有重要意义。因此，发展原位、实时、动态可视化定量检测细胞中活性氧的方法是分析化学面临的挑战和实际需求。本项目将以分子结构和荧光性质可调的BODIPY作为荧光探针母体，设计合成生物兼容性好的系列新型近红外比率型氧化还原可逆荧光探针；研究这些荧光探针与机体内活性氧物种的相互作用机制；通过检测荧光探针与活性氧物种作用时荧光的变化，从中筛选出灵敏度高、线性范围宽、选择性好的荧光探针；结合荧光成像技术，建立原位、实时、动态的可视化检测机体内活性氧物种的新方法，为深入认识活性氧物种的生物学本质及其化学生物学的研究提供技术支撑，同时也将促进生物分析化学和分析检测技术的进步。

本项目以含氰基苯乙烯、吲哚盐、咪唑和香豆素等单元为新型荧光基团，设计并合成了几种反应型荧光探针，以机体内活性氧、活性氮和活性硫物种作为分析对象，利用检测物和荧光探针之间氧化还原反应将荧光探针上的识别基团可逆转化建立检测物浓度-探针荧光信号之间的相关关系，实现了对生物体内活性小分子的荧光检测，并在此基础上将合成的荧光探针用于荧光显微细胞、斑马鱼成像，建立并发展了新型荧光探针用于检测体内生物活性物质的新方法。项目基本上按计划完成，相关成果通过撰写论文和申请国家专利的形式体现，在Angewandte Chemie International Edition, Sensors and Actuators B: Chemical, Talanta等杂志上共发表本研究基金资助的论文51篇（其中SCI论文51篇，IF>3论文45篇），授权国家发明专利1项。培养博士研究生2名，硕士研究生6名。