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 in vitro and in vivo 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 in vitro and in vivo 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.

Boronate-Based Probes for Biological Oxidants: A Novel Class of Molecular Tools for Redox Biology

“…There are numerous reported examples of fluorescent probes designed for the detection of HOCl utilizing the strong oxidizing character of HOCl and specific redox reactions of HOCl (Zhang et al, 2018 ; Wu D. et al, 2019 ). Nonetheless, in most cases, the chemistry of those probes has not been fully characterized, and their biological validation should include an independent and established method of HOCl detection.…”

Malononitrile as the ‘double-edged sword’ of passivation-activation regulating two ICT to highly sensitive and accurate ratiometric fluorescent detection for hypochlorous acid in biological system

“…In addition, we also studied the limits of AI , and observed a good linear relationship I 495 in the range of 0−50 μM. The LOD of AI for HOCl (3σ/slope) [ [25] , [26] , [27] , [28] , [29] , [30] , [31] , [32] , [33] , [34] ] was calculated to be 0.84 μM ( Fig. 4 A).…”

“…On the other hand, the excessive HOCl could cause various diseases such as cardiovascular disease [ 8 ], arthritis [ 9 ], arteriosclerosis [ 10 ] and cancer [ 11 , 12 ]. Therefore, it was of profound significance to realize accurate detection of HOCl in vivo [ [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] , [29] , [30] , [31] , [32] ].…”

Reversible detection of hypochlorite using the deprotonation–protonation strategy: a search for new building blocks

“…4,5 Though the whole process is well regulated, however, a slight over production of hypochlorite may result in the accumulation of ClO À in living systems which eventually generates oxidative stress and hence can cause various neurodegenerative and cardiovascular diseases and cancer. [6][7][8] Over the last few years, a variety of reaction-based fluorescent probes involving irreversible changes around carbon-carbon, [9][10][11][12][13][14] carbon-nitrogen, 15,16 carbonsulphur, 17,18 and carbon-oxygen 19 bonds have been reported for the detection of hypochlorite (Table S1, ESI †). The reaction-based strategy is efficient but hypochlorite induced irreversible chemical changes are introduced in the probe as a result of which the probe is rendered useless after its first application.…”