Rational Design of Near-infrared Fluorescent Probes for Superoxide Anion Radical: Enhancement of Self-stability and Sensitivity by Self-immolative Linker.
Rational Design of Near-infrared Fluorescent Probes for Superoxide Anion Radical: Enhancement of Self-stability and Sensitivity by Self-immolative Linker.
复制标题
DOI：
10.1016/j.freeradbiomed.2021.02.029
复制
发表时间：
2021-03
影响因子：
7.4
通讯作者：
Kaiyun Ji;J. Shan;Xing Wang;Xiaoli Tan;Jingli Hou;Yangping Liu;Yuguang Song
中科院分区：
文献类型：
--
作者：
Kaiyun Ji;J. Shan;Xing Wang;Xiaoli Tan;Jingli Hou;Yangping Liu;Yuguang Song
细胞超氧阴离子自由基(O2·-)的荧光成像对于研究活性氧相关的病理生理过程和药物代谢具有重要意义。然而，由于缺乏合适的探针，该技术的应用还远未达到最大限度。在这项工作中，我们提出了一种新的设计近红外O2·-荧光探针的策略，其中以对甲酚为自焚连接剂，将近红外荧光基团DDAO(9H-1，3-Dichloro-7-hydroxy-9，9-dimethylacridine-2-one)与O2·-敏感基团(即三氟甲磺酸)偶联。自焚连接体的引入有效地提高了这些探针在生理条件下的自稳定性。重要的是，连接基上的吸电子卤素取代基大大提高了探针对O2·-的灵敏度。因此，具有代表性的探针DLS4在很宽的PHS范围内(5.0-8.5)表现出高的自稳定性、高的选择性以及对O2·-的优异灵敏度，检测下限(LOD)为7.3 nm，与O2·-反应时荧光增强720倍。此外，DLS4能够成像PMA刺激的RAW 264.7细胞和HeLa细胞产生O2·-，并且荧光强度与PMA浓度成正比。此外，还用DLS4评价了阿霉素对H9c2细胞的细胞毒作用。本研究为小分子O2·-荧光探针的分子设计提供了一种新的策略，所得到的探针作为研究O2·相关疾病的发生和发展以及不同系统中药物代谢的可靠工具具有巨大的潜力。
Fluorescent imaging of cellular superoxide anion radical (O2•-) is of great significance to investigate reactive oxygen species-related pathophysiological processes and drug metabolism. However, the application of this technique is far away from maximum partially due to the lack of suitable probes. In this work, we propose a new strategy for design of near-infrared (NIR) O2•-fluorescent probes in whichp-cresol is used as a self-immolative linker to conjugate the NIR fluorophore DDAO (9H-1,3-Dichloro-7-hydroxy-9,9-dimethylacridine-2-one) with the O2•--sensing group (i.e., trifluoromethanesulfonate). The introduction of self-immolative linker effectively increases the self-stability of these probes under physiological conditions. Importantly, the electron-withdrawing halogen substituents on the linker greatly enhance the sensitivity of the probes to O2•-. As such, the representative probe DLS4 exhibits high self-stability over a broad range of pHs (5.0–8.5), high selectivity as well as excellent sensitivity to O2•-with a detection limit (LOD) of 7.3 nM and 720-fold fluorescence enhancement upon reaction with O2•-. Moreover, DLS4 enables imaging of O2•-generation in PMA-stimulated RAW 264.7 cells and HeLa cells, and the fluorescence intensities are proportional to the PMA concentrations. In addition, the doxorubicin-induced cytotoxicity of H9c2 cells was also evaluated using DLS4. The present study provides a novel strategy for molecular design of small-molecule O2•-fluorescent probes and the resulting probes show great potential as reliable tools to study the development and progression of O2•--related diseases and drug metabolism in various systems.