Design Principles, Sensing Mechanisms, and Applications of Highly Specific Fluorescent Probes for HOCl/OCl^–

Abstract

Hypochlorous acid/hypochlorite (HOCl/OCl^-), one of the most important reactive oxygen species (ROS), plays vital roles in various physiological and pathological processes. At normal concentrations, OCl^- acts as part of an immune defense system by destroying invasive bacteria and pathogens. However, nonproperly located or excessive amounts of OCl^- are related to many diseases, including cancers. Thus, detection of OCl^- has great importance. Owing to their high sensitivities, selectivities, fast response times, technical simplicities, and high temporal and spatial resolution, fluorescent probes are powerful tools for <i>in vitro</i> and <i>in vivo</i> sensing of target substances. This Account focuses on the development of new chemosensors for detection of OCl^-, which operate by undergoing a chemical reaction with this ROS in conjunction with a change in emission properties. As part of the presentation, we first introduce several important factors that need to be considered in the design of fluorescent chemosensors for OCl^-, including fluorophores, reaction groups, cosolvents, and buffers. Discussion here revolves around the need to select fluorophores that resist oxidation by OCl^-. As well, attention is given to the sensitivities and selectivities of groups in the sensors that react with OCl^- to trigger a fluorescence response. Moreover, well-known reaction groups, which react with highly reactive ROS (hROS), have been redesigned to be specific for OCl^-. In addition, it is pointed out that several cosolvents and buffers such as DMSO and HEPES are not suitable for use in systems for the detection of OCl^- because they are readily oxidized by this ROS. We further discuss recent investigations carried out by us and others aimed at the development of fluorescent probes for <i>in vitro</i> and <i>in vivo</i> detection of OCl^-. These efforts led to the new "dual lock" strategy for designing OCl^- chemosensors as well as several new specific reaction groups such as imidazoline-2-thiones and imidazoline-2-boranes. Probes created using this strategy and the new reacting groups have been successfully applied to imaging exogenous and endogenous OCl^- in live cells and/or tissues. The design concepts and strategies emanating from our studies of fluorescent OCl^- probes have provided insight into the general field of fluorescent probes. Despite the progress made thus far, challenges still remain in developing and applying fluorescent OCl^- probes. For example, more highly specific and sensitive fluorescent OCl^- probes are still in great demand for studies of the biological roles played by OCl^-. Thus, interdisciplinary collaborations of chemists, biologists, and medical practitioners are needed to drive future developments of OCl^- probes for disease diagnosis and drug screening.