Optimizing phenyl selenide-based BODIPYs as fluorescent probes for diagnosing cancer and drug-induced liver injury via cysteine.

Herein, by optimizing phenyl selenide-based BODIPYs, BDP-Se-MOS was obtained, which possessed resistance to ROS and could selectively detect Cys. BDP-Se-MOS could not only discriminate between normal and cancer cells, but also image Cys levels in tumor-bearing mice in real time as well as image the fluctuations of Cys levels in an APAP-induced DILI model.

Three asymmetric BODIPY derivatives as fluorescent probes for highly selective and sensitive detection of cysteine in living cells.

Biothiols are widely involved in various important physiological activities and play a significant role in maintaining redox homeostasis in living organisms. Herein, we designed and synthesized three new asymmetric fluorescent probes (BDP-S-Ph, BDP-S-ENE and BDP-S-R) to discriminate Cys from Hcy/GSH. These probes reacted with Cys to form meso-amino-BODIPYs via SNAr substitution-rearrangement, thereby inducing a fluorescence turn-on effect. Moreover, they could selectively and sensitively detect Cys in solution with low detection limits (50 nM, 28 nM and 87 nM, respectively). Through comparing the response rates of the three probes to Cys, we concluded that the increase of conformational restrictions led to a decrease in probe reactivity. Besides, the sensing mechanism was demonstrated by mass spectrometry. Furthermore, cell experiments indicated that the probes were able to image exogenous and endogenous Cys through green or red channels in living cells.

BODIPY-based Fluorescent Probe for Fast Detection of Hydrogen Sulfide and Lysosome-targeting Applications in Living Cells.

Hydrogen sulfide (H2 S) is recognized as an endogenous gaseous signaling agent in many biological activities. Lysosomes are the main metabolic site and play a pivotal role in cells. Herein, we designed and synthesized two new fluorescent probes BDP-DNBS and BDP-DNP with a BODIPY core to distinguish H2 S. The sensing mechanism is based on the inhibition-recovery of the photo-induced electron transfer (PET) process. Through comparing the responsive behaviors of the two probes toward H2 S, BDP-DNBS showed a fast response time (60 s), low limit of detection (LOD, 51 nM), high sensitivity and selectivity. Moreover, the reaction mechanism was demonstrated by mass spectrometry and fluorescence off-on mechanism was proved by density functional theory (DFT). Significantly, confocal fluorescence imaging indicated that BDP-DNBS was successfully used to visualize H2 S in lysosomes in living HeLa cells.

Detecting Cysteine in Bioimaging with a Near-Infrared Probe Based on a Novel Fluorescence Quenching Mechanism.

Near-infrared (NIR) fluorescent probes are very significant for detecting cysteine in biological systems. Herein, we report a highly selective and sensitive NIR turn-on fluorescent probe (BDP-NIR) based on BODIPY with large Stokes shift (105 nm) for detecting Cys. We clarified the sensing mechanism based on the different thiol-induced SN Ar substitution/rearrangement reaction of the probe with cysteine and homocysteine/glutathione, which leads to the corresponding amino- and thiol-BODIPY dyes with distinct photophysical properties. Moreover, a novel mechanism of fluorescence quenching was demonstrated by density functional theory calculation. The reason for the fluorescence quenching of the probe might be intersystem crossing (from singlet to triplet excited state). Moreover, BDP-NIR had a high linear dynamic range of 0-500 µM, which was promising for detecting cysteine quantificationally. Significantly, BDP-NIR was capable of sensing endogenous cysteine in living cells and in vivo.

Selective Detection and Visualization of Exogenous/endogenous Hypochlorous Acid in Living Cells using a BODIPY-based Red-emitting Fluorescent Probe.

Herein, a red-emitting fluorescent probe DM-BDP-OCl containing a para-DMTC benzyl pyridinium moiety at the meso position of BODIPY as self-immolative portion for the detection of HOCl was designed and synthesized. DM-BDP-OCl exhibited excellent specificity and a fast response for HOCl beyond other ROS/RNS. It was used for the accurately measurable detection of HOCl with a linear range from 0 µM to 50 µM, and the detection limit for HOCl reached 60 nM. Moreover, the probe could directly monitor fluctuations of exogenous and endogenous HOCl in living HeLa and RAW 264.7 cells. This work provided a powerful and convenient imaging tool for probing pathological and physiological actions of HOCl.

BODIPY-Based Fluorescent Probes for Biothiols.

Fluorescent probes for biothiols have aroused increasing interest owing to their potential to enable better understanding of the diverse physiological and pathological processes related to the biothiol species. BODIPY fluorophores exhibit excellent optical properties, which can be readily tailored by introducing diverse functional units at various positions of the BODIPY core. In the present review, the development of fluorescent probes based on BODIPYs for the detection of biothiols are systematically summarized, with emphasis on the preferable detection of individual biothiols, as well as simultaneous discrimination among cysteine (Cys), homocysteine (Hcy), reduced glutathione (GSH). In addition, organelle-targeting probes for biothiols are also highlighted. The general design principles, various recognition mechanisms, and biological applications are elaboratively discussed, which could provide a useful reference to researchers worldwide interested in this area.

Development of Lysosome-Targeted Fluorescent Probes for Cys by Regulating the Boron-dipyrromethene (BODIPY) Molecular Structure.

Our previous discovery suggested that substituents on the 1,7 positions delicately modulate the sensing ability of the meso-arylmercapto boron-dipyrromethene (BODIPY) to biothiols. In this work, the impact of delicate modulations on the sensing ability is investigated. Therefore, 1,7-dimethyl, 3,5-diaryl substituted BODIPY is designed and developed and its conformationally restricted species with a meso-arylmercapto moiety (DM-BDP-SAr and DM-BDP-R-SAr) as selective fluorescent probes for Cys. Moreover, the lysosome-target probes (Lyso-S and Lyso-D) based on DM-BDP-SAr carrying one or two morpholinoethoxy moieties were developed. They were able to detect Cys selectively in vitro with low detection limits. Both Lyso-S and Lyso-D localized nicely in lysosomes in living HeLa cells and exhibited red fluorescence for Cys. Moreover, a novel fluorescence quenching mechanism was proposed from the calculations by density functional theory (DFT). The probes may go through intersystem crossing (from singlet excited state to triplet excited state) to result in fluorescence quenching.

A water-soluble BODIPY-based fluorescent probe for rapid and selective detection of hypochlorous acid in living cells.

We designed and synthesized 4,4-di-(4'-methylmercaptophenoxy)-8-(N-methylpyridinium-2-yl)-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene (probe 1) as a water-soluble BODIPY derivative for rapid and selective detection of hypochlorous acid. The pyridinium-2-yl linked at the meso position of BODIPY core was used to maintain highly fluorescent nature and to increase water solubility. Methylmercaptophenoxy was selected as responsive site installed on the boron atom (to replace the fluorine atom) and induced the photoinduced electron-transfer (PeT) effect to quench the fluorescence of BODIPY. The probe exhibited 83.9 µg mL-1 solubility in PBS (10 mM, pH 7.4), and possessed very low fluorescence (Φf = 0.0013). Upon addition of HClO, the probe could display a distinct response in 1 min and generate marked fluorescence enhancement by 100-fold due to the oxidation of thioether into sulfoxide to terminate PeT process. A limit of detection of 53 nM was calculated for HClO in the linear response range from 0 µM to 10 µM, and the probe was successfully applied to image HClO in living cells.

BODIPY-based turn-on fluorescent probes for cysteine and homocysteine.

Cysteine (Cys), homocysteine (Hcy), and glutathione (GSH) are interconnected and play essential roles in many biological processes. It is significant importance to detect these thiols for investigating their functions in cells and disease diagnosis. In this work, we have designed and synthesized two novel BODIPY-based turn-on fluorescent probes (BDP-Ph and BDP-R-Ph) carrying 4-methoxythiophenol moiety at meso position as good leaving group for highly selective detection of Cys and Hcy. Furthermore, the probes have been successfully applied to detect intracellular Cys and Hcy by fluorescent imaging in living cells.