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Three-Photon AIE Pt(II) Complexes as Cysteine-Targeting Theranostic Agents for Tumor Imaging and Chemotherapy
Abstract
Herein, we have synthesized a series of three-photon fluorescent Pt(II) complexes targeting a tumor-associated biothiol, cysteine (Cys), which allows it to be detected without any interference from other intracellular proteins. We focused on how to significantly improve the fluorescence response of Cys via regulating the recognition units in probes. The reaction of K2PtCl4 with L-CH3 or L-COOEt in DMSO solution gave Lyso-Pt-CH3 and Lyso-Pt-COOEt, respectively, which present four-coordinated square-planar geometries in mononuclear structures. Lyso-Pt-CH3 consists of a Cys aptamer labeled with typical aggregation-induced emission (AIE) characteristics, which shows strong three-photon absorption cross section (3PA) only in the presence of Cys. It was found that Lyso-Pt-CH3 displayed a perfect signal-to-noise ratio for imaging lysosomes and for rapid detection of Cys. Using Lyso-Pt-CH3, Cys-related cellular mechanisms were proposed. We confirm that cystine (Cyss) could be absorbed in cells through cystine/glutamate antiporters (system xc-) and is then converted to Cys under the effect of enzymes. All of these suggest that Lyso-Pt-CH3 might be a potential candidate as a simple and straightforward biomarker of lysosome-related Cys in vitro. Lyso-Pt-CH3 can effectively identify tumor tissues with excessive levels of Cys. Lyso-Pt-CH3 also showed excellent antitumor activity than cisplatin. This work provides a novel strategy for the rational design of controllably activated and Cys-targeted Pt(II) anticancer prodrugs for clinical diagnosis and treatment.
Cysteine is an important regulator of redox processes. Due to the nucleophilic and oxidative sensitivity, cysteine residues in proteins can be oxidized by intracellular reactive oxygen species (ROS), which can lead to protein structural and functional changes. Hence, the development of fluorescent probes to image cysteine and cysteine oxidation is of great significance for the study of redox homeostasis in living system. In this review, the development of fluorescent probes for imaging cysteine and cysteine oxidation was summarized. Moreover, we further analyzed defects of the reported fluorescent probes and made suggestions for the future development of fluorescent probes. We expect that this review can not only provide a deeper understanding of the role of cysteine and cysteine oxidation in oxidative stress, but also broaden the application of fluorescent probes in imaging cysteine and cysteine oxidation.
The early detection of tumors and precancerous conditions is vital for cancer diagnosis. Advances in fluorescence microscopic techniques and materials synthesis processes have revolutionized biomarker detection and image-guided cancer surveillance. In particular, novel materials-based diagnostic tools and innovative therapies have facilitated a precise understanding of biological processes at the molecular level. This critical review presents an overview of bioimaging probes, including functionalized chromophoric systems, non-functionalized chromophoric systems, and nanoscale biosensors. Technical challenges and future directions related to these approaches are considered. Published under an exclusive license by AIP Publishing. https://doi.
The discovery of the aggregation-induced emission (AIE) phenomenon in certain classes of organic molecules has revolutionized our understanding of the photoluminescence properties of materials. This breakthrough has opened up new...
The implementation of early cancer detection benefits the treatment outcomes with remarkably improved survival rate through the detection of rare circulating biomarkers in body fluids. Spectroscopic technologies play a crucial role in sensitive biomarker measurements by outputting extremely strong signals. In particular, the aggregation enhanced fluorescence and Raman technologies feature the detection of targets down to single-molecule level, thereby demonstrating the great promise of early cancer detection. In this review, we focus on the aggregation-induced emission (AIE) and aggregation-related surface-enhanced Raman scattering (SERS) spectroscopic strategies for detecting cancer biomarkers. We discuss the AIE and SERS based biomarker detection using target-driven aggregation as well as the aggregated nanoprobes. Furthermore, we deliberate on the progress of developing AIE and SERS integrated platforms. Ultimately, we put forth the potential challenges and perspectives on the way to use these two spectroscopic technologies in clinical settings. It is expected this review can inspire the design of AIE and SERS integrated platform for highly sensitive and accurate cancer detection.
Specific intervention of senescent cells (SnCs) is emerging as a powerful means to counteract aging and age‐related diseases. Canonical methods are generally designed to target SnC‐associated signaling pathways, which are however dynamically changing and highly heterogeneous in SnCs, significantly limiting the effectiveness. Here, we present a tailor‐made molecular prodrug targeting lysosome dysfunction, a unique feature shared by virtually all types of SnCs. The prodrug comprises three modules all targeting the altered lysosomal programs in SnCs, namely, a recognizing unit towards the elevated lysosome content, a linker cleavable by the activated lysosomal enzyme, and a lysosomotropic agent targeting the increased lysosomal membrane sensitivity. This spatially confined design enables killing broad‐spectrum SnCs, with high specificity over non‐SnCs. Along with in vivo benefits, this work offers a way to significantly expand the applicability of senotherapy in a wide range of diseases.
A remaining challenge in the treatment of glioblastoma multiforme (GBM) is surmounting the blood-brain barrier (BBB). Such challenge prevents the development of efficient theranostic approaches that combine reliable diagnosis with targeted therapy. In this study, we developed brain-targeted near-infrared Ⅱb (NIR-Ⅱb) aggregation-induced emission (AIE) nanoparticles via rational design that involved twisting the planar molecular backbone with steric hindrance. The resulting nanoparticles can balance competing responsiveness demands for radiation-mediated NIR fluorescence imaging at 1550 nm and non-radiation NIR photothermal therapy (NIR-PTT). The brain targeting peptide apolipoprotein E peptide (ApoE) is grafted onto these nanoparticles (termed as ApoE-Ph NPs) to target glioma and promote efficient BBB traversal. A long imaging wavelength 1550 nm band-pass filter was utilized to monitor the in vivo biodistribution and accumulation of our nanoparticles in a model of orthotopic glioma, which overcame previous limitations in wavelength range and equipment. The results demonstrate that our ApoE-Ph NPs have a higher PTT efficiency and significantly enhanced survival of mice bearing orthotopic GBM with moderate irradiation (0.5 W cm–2). Collectively, our work highlights the smart design of a brain-targeted NIR-Ⅱ AIE theranostic approach that opens new diagnosis and treatment options in the photonic therapy of GBM.
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Chromophores that exhibit aggregation-induced emission (i.e., aggregation-induced emission luminogens [AIEgens]) emit intense fluorescence in their aggregated states, but show negligible emission as discrete molecular species in solution due to the changes in restriction and freedom of intramolecular motions. As solvent-swollen quasi-solids with both a compact phase and a free space, gels enable manipulation of intramolecular motions. Thus, AIE-active gels have attracted significant interest owing to their various distinctive properties and promising application potential. Herein, a comprehensive overview of AIE-active gels is provided. The fabrication strategies employed are detailed, and the applications of AIEgens are summarized. In addition, the gel functions arising from the AIE moieties are revealed, along with their structure-property relationships. Furthermore, the applications of AIE-active gels in diverse areas are illustrated. Finally, ongoing challenges and potential means to address them are discussed, along with future perspectives on AIE-active gels, with the overall aim of inspiring research on novel materials and ideas.
The unique advantages and the exciting application prospects of AIEgens have triggered booming developments in this area in recent years. Among them, stimuli-responsive AIEgens have received particular attention and impressive progress, and they have been demonstrated to show tremendous potential in many fields from physical chemistry to materials science and to biology and medicine. Here, the recent achievements of stimuli-responsive AIEgens in terms of seven most representative types of stimuli including force, light, polarity, temperature, electricity, ion, and pH, are summarized. Based on typical examples, it is illustrated how each type of systems realize the desired stimuli-responsive performance for various applications. The key work principles behind them are ultimately deciphered and figured out to offer new insights and guidelines for the design and engineering of the next-generation stimuli-responsive luminescent materials for more broad applications.
BODIPYs are renowned fluorescent dyes with strong and tunable absorption in the visible region, high thermal and photo-stability and exceptional fluorescence quantum yields. Transition metal complexes are the most commonly used triplet photosensitisers, but, recently, the use of organic dyes has emerged as a viable and more sustainable alternative. By proper design, BODIPY dyes have been turned from highly fluorescent labels into efficient triplet photosensitizers with strong absorption in the visible region (from green to orange). In this perspective, we report three design strategies: (i) halogenation of the dye skeleton, (ii) donor-acceptor dyads and (iii) BODIPY dimers. We compare pros and cons of these approaches in terms of optical and electrochemical properties and synthetic viability. The potential applications of these systems span from energy conversion to medicine and key examples are presented.
Abnormal concentrations of biothiols such as cysteine, homocysteine and glutathione are associated with various major diseases. In biological systems, the structural similarity and functional distinction of these three small molecular thiols has not only required rigorous molecular design of the fluorescent probes used to detect each thiol specifically, but it has also inspired scientists to uncover the ambiguous biological relationships between these bio-thiols. In this minireview, we will discuss the evolution of small organic molecular fluorescent probes for the detection of thiols over the past 60 years, highlighting the potent methodologies used in the design of thiol probes and their particular applications in the semi-quantification of cellular thiols and real-time labelling. At the same time, the present challenges that limit their further application will be discussed. We hope that this minireview will promote future research to enable deeper insight into the crucial role of thiols in biological systems.
Crystallization-induced photoluminescence weakening was recently revealed in ultrasmall metal nanoparticles. However, the fundamentals of the phenomenon are not understood yet. By obtaining conformational isomer crystals of gold nanoclusters, we investigate crystallization-induced photoluminescence weakening and reveal that the shortening of interparticle distance decreases photoluminescence, which is further supported by high-pressure photoluminescence experiments. To interpret this, we propose a distance-dependent non-radiative transfer model of excitation electrons and support it with additional theoretical and experimental results. This model can also explain both aggregation-induced quenching and aggregation-induced emission phenomena. This work improves our understanding of aggregated-state photoluminescence, contributes to the concept of conformational isomerism in nanoclusters, and demonstrates the utility of high pressure studies in nanochemistry.
One of the most critical factors for the survival of glioblastoma (GBM) patients is the precision diagnosis and the tracking of the treatment progress. At the moment, there are various sophisticated and specific diagnostic procedures being used, but there are relatively few simple diagnosis methods. This work introduces a sensing probe based on a turn-on type fluorescence response that can measure cysteine (Cys) level in the human-derived cells, as well as within on-site human clinical biopsy samples, which recognize Cys as a new biomarker of GBM. The Cys-initiated chemical reactions of the probe cause a significant fluorescence response with high selectivity, high sensitivity, a fast response time, and a two-photon excitable excitation pathway, which allows the imaging of GBM in both mouse models and human tissue samples. The probe can distinguish the GBM cells, as well as disease sites in clinical samples from individual patients. Besides, the probe has no short or long-term toxicity and immune response. The present findings hold promise for application of probe to a relatively simple and straightforward following of GBM at clinical sites.
To probe the regulatory roles of cysteine (Cys) in cancel cell survival, a highly selective and sensitive fluorescent Cys probe SiR was developed by employing a novel “lock and key” strategy, which allows Cys to be detected without any interference or probe consumption caused by intracellular high concentration of glutathione (GSH). Using SiR, we confirmed that inhibiting cystine (Cys2) transporter system xc⁻ to deplete intracellular Cys is more efficient than inhibiting glutamate–cysteine ligase GCL to deplete intracellular GSH for sensitizing cancer cells to chemotherapy. Moreover, with the probe, a possible self-protection mechanism of cancer cells was indicated: when extracellular Cys sources are blocked, cancer cells could still survive by multidrug resistance protein transporter (Mrp1)-mediated export of intracellular GSH/GSSG as sources to supply intracellular Cys for resisting detrimental oxidative stress. Based on this finding, we further confirmed that abrogating the self-protection mechanism is an even more efficient strategy for sensitizing cancer cells to chemotherapy.
Cysteine (Cys) is important in biosynthesis, detoxification, and metabolism. The selective detection of Cys over structurally similar homocysteine (Hcy) or glutathione (GSH) remains an immense challenge. Although there are many methods for detecting Cys, photoluminescence (PL) and electrochemiluminescence (ECL) techniques are well-suited for clinical diagnostics and analytical technology because of their high sensitivities. Herein, we report PL and ECL dual-channel sensors using cyclometalated iridium(III) complexes for the discrimination of Cys from Hcy and GSH. The sensors react with cysteine preferentially because of kinetic differences in intramolecular conjugate addition/cyclization, enabling phosphorescence enhancement and ECL decrease in the blue-shifted region. Sensor 1 shows ratiometric PL turn-on and ECL turn-off for Cys. In addition, unique ECL-enhancing behavior of sensor 1 toward GSH enables discrimination between Cys and GSH. Sensor 1 was successfully applied to the detection of Cys in human serum by the ECL method. We demonstrate the first case of a Cys-selective PL and ECL dual-channel chemodosimetric sensor based on cyclometalated iridium(III) complexes and expect that the rational design of efficient PL and ECL dual-channel sensors will be useful in diagnostic technology.
Cisplatin‐based chemotherapeutic regimens are frequently used for treatments of solid tumors. However, tumor cells may have inherent or acquired cisplatin resistance, and the underlying mechanisms are largely unknown. We performed genome‐wide screening of genes implicated in cisplatin resistance in A375 human melanoma cells. A substantial fraction of genes whose disruptions cause cisplatin sensitivity or resistance overlap with those whose disruptions lead to increased or decreased cell growth, respectively. Protein translation, mitochondrial respiratory chain complex assembly, signal recognition particle‐dependent cotranslational protein targeting to membrane, and mRNA catabolic processes are the top biologic processes responsible for cisplatin sensitivity. In contrast, proteasome‐mediated ubiquitin‐dependent protein catabolic process, negative regulations of cellular catabolic process, and regulation of cellular protein localization are the top biologic processes responsible for cisplatin resistance. ZNRF3, a ubiquitin ligase known to be a target and negative feedback regulator of Wnt–β‐catenin signaling, enhances cisplatin resistance in normal and melanoma cells independently of β‐catenin. Ariadne‐1 homolog (ARIH1), another ubiquitin ligase, also enhances cisplatin resistance in normal and melanoma cells. By regulating ARIH1, neurofibromin 2, a tumor suppressor, enhances cisplatin resistance in melanoma but not normal cells. Our results shed new lights on cisplatin resistance mechanisms and may be useful for development of cisplatin‐related treatment strategies.—Ko, T., Li, S. Genome‐wide screening identifies novel genes and biological processes implicated in cisplatin resistance. FASEB J. 33, 7143–7154 (2019). www.fasebj.org
Although fluorescence imaging diagnosis of the differences between cancer cells and normal cells by the targeting ligand-based fluorescent probes is useful to classify patients towards personalized therapy, using the differences to diagnose a wide range of cancers is often not possible due to the genetic or phenotypic heterogeneity of cancer cells. In this work, a 2-(diphenylphosphino)phenol-functionalized pyronin POP was presented as a dual-channel fluorescence agent for diagnosing a wide range of cancer cell types. The agent could efficiently penetrate cancer cell, rather than normal cell, membranes by active transport of organic-anion transporting polypeptide (OATP) transporters overexpressed in many types of cancer cells, and then is activated by intracellular cysteine (Cys) and glutathione (GSH) to produce green-emission aminopyronin NP and red-emission thiopyronin SP, thereby enabling its use in dual-channel fluorescence diagnosis of a wide range of cancer cells with excellent contrast. Crucially, POP also displays the ability of dual-channel fluorescence diag¬nosis of cancer tissues from tumour xenograft models of mice and harvested surgical specimens of patients, thus holding great potential for the clinical applications.
Abnormal concentrations of Cys have been reported to be implicated in various health problems including cancer, neuropathy, and cardiomyopathy. We present a novel two-photon fluorescent probe for the specific recognition of cysteine over homocysteine and glutathione, and the bioapplication of this probe for the imaging of live cancerous cells and thick tissues.
The dynamic chemical diversity of elements, ions and molecules that form the basis of life offers both a challenge and an opportunity for study. Small-molecule fluorescent probes can make use of selective, bioorthogonal chemistries to report on specific analytes in cells and in more complex biological specimens. These probes offer powerful reagents to interrogate the physiology and pathology of reactive chemical species in their native environments with minimal perturbation to living systems. This Review presents a survey of tools and tactics for using such probes to detect biologically important chemical analytes. We highlight design criteria for effective chemical tools for use in biological applications as well as gaps for future exploration.
Cancer is a health threat worldwide, and it is urgent to develop more sensitive cancer detection methods. Herein, a polarity-sensitive cell membrane probe (named COP) was developed for detecting cancer cells and tumors sensitively and selectively at the cell membrane level. The probe shows a strong polarity-dependent fluorescence and excellent cell membrane targeting ability to visualize cell membrane with red fluorescence with a non-washing process. Notably, COP can selectively light up the tumor cell membranes, which reveals that cancer cell membranes have lower polarity than normal cell membranes. The giant unilamellar vesicle model and cell imaging studies proved this. Moreover, COP can effectively and selectively light up tumors. Overall, this work demonstrates that the polarity of the tumor cell membrane is quite different to normal cell membranes, and based on this, sensitive membrane probes can be developed to selectively visualize cancer cells and tumors, which opens up a new way for tumor diagnosis at the cellular level.
Lysosome is one of the most crucial organelles in living systems. Lysosomal viscosity is an important microenvironment parameter in lysosomes, which is closely related to the occurrence and development of many diseases, including cancer. To monitor the highly dynamic lysosomes and the lysosomal viscosity changes, we developed a versatile high-performance fluorescent probe DCMP in this work. DCMP shows stronger fluorescence under acidic conditions or in solutions with higher viscosity, especially with high sensitivity to the change of viscosity. More importantly, DCMP can use a low dose (100 nM) and can target lysosomes and image lysosomes with high signal-to-noise ratio and high fidelity. It can also track the lysosomal motility and viscosity changes in real-time, and effectively monitor the interaction between lysosomes and damaged organelles. We also found that DCMP can be successfully used to selectively and sensitively light up cancer cells and cancer tissues. All the results show that this new probe has great potential not only in lysosomal high-fidelity imaging, but also in cancer detection.
Both glutathione (GSH) and viscosity play an important role in mitochondria. They are closely related to various physiological and pathological processes and are important biomarkers in cancer analysis. Herein, we developed the first dual responsive fluorescence probe MGV that can simultaneously detect mitochondrial GSH and viscosity. MGV is smart and shows a selective ratiometric response to GSH at 535/650 nm with a significant increase of green fluorescence. MGV can also show a distinct red fluorescence enhancement at 627 nm as viscosity increases. In addition, MGV has low cytotoxicity and good mitochondrial-targeting ability. With these features, MGV was successfully applied to image mitochondrial GSH at dual fluorescence channels and track mitochondrial viscosity changes on the red fluorescence channel. With MGV, the formation of mitochondrial bleb vesicles was observed with nystatin stimulation, and during the Cisplatin-induced apoptosis, both the level of GSH and the viscosity showed an increase. Finally, based on the dual-response to GSH and viscosity, MGV was successfully used for dual-channel imaging of cancer cells/tumors. Overall, this work provided a smart probe for GSH and viscosity and new insights/methods for apoptosis and tumor imaging.
In this work, by installing a free-rotating benzene ring to suppress the intermolecular π-π stacking effect in the aggregated state, a naphthalimide-derived hypochlorite (ClO-) fluorescent probe, Probe A, with the aggregation-caused quenching (ACQ) effect was successfully transformed into Probe B possessing typical aggregation-induced emission (AIE) characteristics. The experimental results indicated that Probe B with good selectivity and a low detection limit (LOD) of 0.02 µM can also exhibit a significant ratiometric fluorescence color change from cyan to dark blue within 2 min in a nearly pure water solvent system after the addition of ClO-. Finally, by virtue of the good photophysical properties and ClO- sensing performance, Probe B and the Probe B loaded portable test paper were successfully applied to live cell imaging and the naked eye recognition of ClO-, respectively.
Platinum-based complexes are one of the most successful chemotherapeutic agents having a significant ground in cancer chemotherapy despite their side effects. During the past few decades, Ru(II) complexes have been emerging as efficient alternatives owing to their promising activities against platinum-resistant cancer. The pathway of action, lipophilicity, and cytotoxicity of a Pt or Ru complex may be tuned by varying the attached ligands, the coordination mode, and the leaving group. In this work, we report a family of Pt(II) and Ru(II) complexes (1-5) of three N,O and N,N donor-based trimethoxyanilines containing Schiff bases with the general formula [PtII(L)(DMSO)Cl], [RuII(L)(p-cymene)Cl], [RuII(L)(p-cymene)Cl]+, and [PtII(L)Cl2]. All of the complexes are characterized by different analytical techniques. 1H NMR and electrospray ionization mass spectrometry (ESI-MS) data suggest that the N,O-coordinated Pt(II) complexes undergo slower aquation compared to the Ru(II) analogues. The change of the coordination mode to N,N causes the Ru complexes to be more inert to aquation. The N,O-coordinating complexes show superiority over N,N-coordinating complexes by displaying excellent in vitro antiproliferative activity against different aggressive cancer cells, viz., triple-negative human metastatic breast adenocarcinoma MDA-MB-231, human pancreatic carcinoma MIA PaCa-2, and hepatocellular carcinoma Hep G2. In vitro cytotoxicity studies suggest that Pt(II) complexes are more effective than their corresponding Ru(II) analogues, and the most cytotoxic complex 3 is 10-15 times more toxic than the clinical drugs cisplatin and oxaliplatin against MDA-MB-231 cells. Cellular studies show that all of the N,O-coordinated complexes (1-3) initiate disruption of the microtubule network in MDA-MB-231 cells in a dose-dependent manner within 6 h of incubation and finally lead to the arrest of the cell cycle in the G2/M phase and render apoptotic cell death. The disruption of the microtubule network affects the agility of the cytoskeleton rendering inhibition of tyrosine phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2), a key step in angiogenesis. Complexes 1 and 2 inhibit VEGFR2 phosphorylation in a dose-dependent fashion. Among the Pt(II) and Ru(II) complexes, the former displays higher cytotoxicity, a stronger effect on the cytoskeleton, better VEGFR2 inhibition, and strong interaction with the model nucleobase 9-ethylguanine (9-EtG).
Epilepsy is characterized by oxidative stress in the brain. As the crucial reductive biothiol, cysteine (Cys) directly regulates the occurrence of oxidative stress in the brain. Observations suggest that the decreased cysteine in plasma could potentially serve as a redox biomarker in temporal lobe epilepsy. However, due to the complexity of the brain and lack of appropriate in-situ detecting tools, the concentration change and regulation of Cys in epileptic brains remain unclear. Here, we report a near-infrared imaging probe (named Mito-CP) for tracking endogenous Cys in brains of pentylenetetrazole (PTZ)-induced epileptic seizures with high sensitivity and selectivity. Using this probe, we achieved an in-situ visualization of the increased Cys in PC12 cells under dithiothreitol stimulation. In addition, Mito-CP was able to real-time monitor Cys fluctuation in lipopolysaccharide-mediated oxidative stress in zebrafish. Ultimately, we directly visualized the precipitous reduction of Cys in epileptic brains for the first time. Mito-CP also revealed the fluctuation of Cys in epileptic brains during the treatment by an antiepileptic drug, curcumin. This work provides an effective tool for Cys imaging in brains and will help to expand the understanding of the pathogenesis of epilepsy.
A yolk−shell structured Au@Ag@mSiO2 probe was fabricated by coating a layer of mesoporous silica on the surface of Au@Ag core-shell nanosphere, followed by partially removing the Ag shell. Mirror symmetrical chiral signals at ~258 nm in the UV region were observed for the probe upon coupling with cysteine enantiomers on the surface of Au@Ag. The intensity of the CD signal of the probe is enhanced with increasing L/D-Cys concentration, allowing the quantitatively determination of the cysteine enantiomers. The developed method exhibits an excellent linear relationship between the CD signal and L-Cys concentration ranging from 10 μM to 90 μM with a limit of detection of 8.5 μM. In the presence of Cu2+, the CD signal of probe was weakened due to the oxidation of L-cysteine to L-cystine catalyzed by Cu2+. Based on this phenomenon, a new strategy for detection of Cu2+ can be established. Under optimized condition, the CD signal decreases linearly with the log of the concentration of Cu2+ in the range from 1 to 250 nM with a detection limit of 0.1 nM.
Cysteine (Cys) and homocysteine (Hcy) are essential for maintaining the cellular redox homeostasis and play critical roles in patho-logical and physiological processes. The development of Cys/Hcy-specific responsive fluorescent probe that are independent to the surrounding environment, equipment and abundant endogenous GSH is critical to accurately investigate the roles of Cys/Hcy in living biological systems. In this work, a novel ratiometric and mitochondria-anchored fluorescence chemosensor, PYR, was con-structed on the basis of 4-methylphenol-substituted pyronin fluorophore. The probe exhibited ratiometric fluorescence emission (F540 nm/F620 nm) for the detection of Cys/Hcy with high selectivity, sensitivity (Cys, 22 nM; Hcy, 23 nM), rapid response (Cys, 5 min) and a merit enhancement of ratio fluorescent signal (Cys, 163-fold; Hcy, 125-fold). The probe showed excellent membrane permeability and was applied to visualize mitochondrial biothiols in living cells under H2O2-induced redox imbalance, kidney tis-sues with a penetration depth of 100 μm and Daphnia magna model for the first time. The results demonstrate that PYR will pro-vide a promising platform for the diagnosis of thiol-related diseases.
A water-soluble turn-on fluorescent probe has been rationally designed and synthesized to distinguish Cys from Hcy and GSH in less than five seconds. It has high sensitivity and strong anti-interference...
Depression is characterized by oxidative stress in the brain. As the crucial reductive biothiol, cysteine (Cys) directly regulate the occurrence of oxidative stress in brain. Despite its significance, the precise exploration of Cys in mouse brains remains a challenge, primarily owing to the limitations of Cys-monitoring tools, especially the interference from unavoidable reaction with other biothiols. Thus, we developed a novel two-photon fluorescence probe for Cys based on a new specific recognition site, thiobenzoate. Encountering Cys, the carbon-sulfur double bond in the probe formed a stable five-membered ring via the selective nucleophilic addition reaction, triggering a remarkable fluorescence increase. Notably, this reaction can’t occur with other biothiols, which afford the probe unprecedented selectivity to Cys. With two-photon excitation at 754 nm, we achieved in situ visualization of the increased Cys in PC12 cells under dithiothreitol stimulation. Furthermore, we directly visualized the precipitous reduction of Cys in the brains of mice with depression phenotypes for the first time. This work opens up new vistas for Cys imaging and expands the understanding of pathogenesis of depression.
Herein we utilize the similar though divergent nucleophilic properties of cysteine, homocysteine, and glutathione to achieve the selective detection of cysteine under mildly acidic conditions. This enables the specific in situ detection of lysosomal cysteine. Employing time dependent fluorescent imaging of probe-labeled A549 cells, we demonstrate that dexamethasone-induced apoptosis is not dependent on lysosomal cysteine. This methodology can thus produce useful information about pathogenesis associated with cysteine and lysosomes.
Coumarinocoumarin, one of the coumarin derivatives, is easy to synthesize and have rich modification sites. The large conju-gate plane of coumarinocoumarin make it has more excellent optical property than conventional coumarin, for example, the two-photon fluorescence property. So, the coumarinocoumarin-based probe (CCy) has been designed and synthesized, which is the first lysosomal targeting fluorescent biosensor for cysteine. This probe was prepared by a 3-step procedure as a latent fluorescence probe to achieve high sensitivity and fluorescence turn-on response toward cysteine (Cys) over homocys-teine (Hcy), glutathione (GSH) and other various natural amino acids under physiological conditions. Upon addition of Cys to the solution of CCy, remarkable enhancement on 520 nm of fluorescence spectra can be monitored. This probe was then successfully used for fluorescence imaging of Cys in mice organ tissues and HeLa cells, and quantitative determination had been achieved within a certain range, which proved the permeability of CCy. The concentration of Cys in animal serum was measured and the viability exceeded 80%, indicating that CCy can be a biocompatible and rapid probe for Cys in vivo. Simul-taneously, its ability to detect Cys in lysosome has been validated by its lysosomal targeting.
This work describes the synthesis, characterization and biological evaluation of three platinum complexes of the type [Pt(DMSO)(L)Cl]Cl, in which L represents a fluoroquinolone, namely, ciprofloxacin (cpl), ofloxacin (ofl), or sparfloxacin (spf). The new complexes were characterized by elemental analysis, high-resolution mass spectrometry (HRESIMS) and1H,13C and195Pt NMR (nuclear magnetic resonance). The spectral data suggest that the fluoroquinolones act as bidentate ligands coordinated to Pt(II) through the nitrogen atoms of the piperazine ring. Microbiological assays against wild type Mycobacterium tuberculosis (ATCC 27294) showed that all complexes have been very potent, exhibiting antitubercular potency at concentrations <2 μM, although none of the complexes presented higher potency than established anti-TB drugs. As to the resistant strains, the complex with sparfloxacin, [Pt(DMSO)(spf)Cl]Cl exhibited the best potential against most Mycobacterium tuberculosis clinical isolates. The cytotoxicity of these compounds was also evaluated in three breast cell lines: MCF-10 (a healthy cell), MCF-7 (a hormone responsive cancer cell) and MDA-MB-231 (triple negative breast cancer cell). In both tumor cell lines, [Pt(DMSO)(spf)Cl]Cl was more active and more selective than cisplatin. Flow cytometry analysis revealed that [Pt(DMSO)(spf)Cl]Cl induced late apoptotic cell death in MDA-MB-231 cells.
Cysteine (Cys), as an important bio-thiol, plays a major role in many physiological processes like protein synthesis, detoxification and metabolism, and also is closely associated with a variety of diseases; thus the design of novel highly selective and sensitive near-infrared (NIR) fluorescent probes for Cys detection in vivo is of great significance. Herein, we report a selective and sensitive NIR turn-on fluorescent probe (CP-NIR) with large Stokes shift for detecting Cys in vivo. Upon addition of Cys to the solution of the probe, it’s absorption wavelength shifts from 550 to 600 nm, accompanying with an obvious enhancement of NIR fluorescence emission centering around 760 nm. This Michael-addition-reaction based probe shows a large Stokes shift (160 nm), low detection limit (48 nM), fast response time and low toxicity. Moreover, this novel NIR probe with good cell permeability was successfully applied to monitoring endogenous Cys in living cells and in a mouse model.
The development of effective molecular probes to detect and image the levels of oxidative stress in cells remains a challenge. Herein we report the design, synthesis and preliminary biological evaluation of a novel optical probe to monitor oxidation of thiol groups in cysteine-based phosphatases (CBPs). Following orthogonal protecting approaches we synthesised a new vanadyl complex designed to bind to CBPs. This complex is functionalised with a well-known dimedone derivative (to covalently trap sulfenic acids, SOHs) and a coumarin-based fluorophore for optical visualization. We show that this new probe efficiently binds to a range of phosphatases in vitro with nanomolar affinity. Moreover, preliminary flow cytometry and microscopy studies in live HCT116 cells show that this probe can successfully image cellular levels of sulfenic acids – one of the species resulting from protein oxidative damage.
A dual mode fluorescent probe, which is based on an integration of fluorescein and coumarin fluorophores, was developed for the discrimination of Cys from Hcy and GSH. This probe (2) shows the advantage of quick reaction (5 min) with Cys, resulting in a strong fluorescence turn-on response when excited at 450 nm. Notably, it also demonstrates the ratiometric fluorescence property while excited by a shorter wavelength (332 nm). All of results suggest probe 2 has a high selectivity toward Cys even in the presence of other amino acids, cations and anions. The detection limit of Cys was calculated as 0.084 μM, which was much lower than the intracellular concentration. 1H NMR, MS and DFT calculation were used to reveal the detection mechanism further. Finally, this low cytotoxic probe was successfully applied in bioimaging within HepG2 cells.
The design of small molecule fluorescent sensors is currently a popular research field, which stands to contribute much to the understanding of biological systems. However, it is essential that efforts be better directed, to maximize the utility of such systems. This Sensor Issues article discusses the current challenges and opportunities in the field, highlighting the need for better collaboration between chemists and biological researchers in this domain.
A long lifetime iridium(iii) complex chemosensor for cysteine detection has been synthesized. The luminescence signal of could be recognized in strongly fluorescent media through time-resolved emission spectroscopy (TRES). The detection of cysteine in a living zebrafish was demonstrated.
The serendipitous discovery of the anticancer drug cisplatin cemented medicinal inorganic chemistry as an independent discipline in the 1960s. Luminescent metal complexes have subsequently been widely applied for sensing, bio-imaging, and in organic light-emitting diode applications. Transition-metal complexes possess a variety of advantages that make them suitable as therapeutics and as luminescent probes for biomolecules. It is thus highly desirable to develop new luminescent metal complexes that either interact with DNA through different binding modes or target alternative cellular machinery such as proteins as well as to provide a more effective means of monitoring disease progression. In this Review, we highlight recent examples of biologically active luminescent metal complexes that can target and probe a specific biomolecule, and offer insights into the future potential of these compounds for the investigation and treatment of human diseases.
Luminescent transition metal complexes are currently revolutionizing many areas of photochemistry and photophysics. In particular, they are proving useful as molecular probes and sensors. We discuss the design consideration in producing useful sensors and probes. As we show, complexes are amenable to rational design. Applications of inorganic complexes to a variety of sensor technologies are discussed. In addition, problem areas such as sensor–support interactions are covered.
A series of the new ruthenium(II) complexes with different number of aldehyde groups have been synthesized and characterized for the simple and selective sensing of homocysteine (Hcy) and cysteine (Cys). The reaction of these ruthenium(II) complexes with Hcy and Cys afforded thiazinane or thiazolidine derivatives which resulted in the obvious changes in the UV-visible spectra and strong enhancement of the luminescence intensity of the system. The luminescence enhancement of [Ru(dmb)(2)(L2)](2+) (dmb: 4,4'-dimethyl-2,2'-bipyridine) showed a good linearity in the concentration of 4.2-350 μM and 6-385 μM with the detection limits of 0.3 μM and 1 μM for Hcy and Cys, respectively. The absorption and emission bands from metal-to-ligand charge transfer transition in the visible region and the large Stokes shift of the ruthenium(II) complex chromophore made it suitable for biological applications.
Platinum-containing drugs are widely used to treat cancer in a variety of clinical settings. Their mode of action involves the formation of DNA adducts, which facilitate apoptosis in cancer cells. Cisplatin binds to the N7 position of the purine DNA bases forming intrastrand cross-links between either two adjacent guanines [cis-Pt(NH(3))(2)d(pGpG), 1,2-GG] or an adjacent adenine and guanine [cis-Pt(NH(3))(2)d(pApG), 1,2-AG)]. The cytotoxic efficacy for each of the different types of DNA adducts and the relationship between adduct levels in tumor cells and blood are not well understood. By using these Pt-containing adduct species as biomarkers, information on a patient's response to chemotherapy would be directly related to the mode of action of the drug. This type of analysis requires the most sensitive and specific methods available, to facilitate detection limits sufficient to measure the DNA adduct in the limited sample quantities available from patients. This was achieved in the current study by coupling a highly specific enzyme-based adduct isolation method with a sensitive detection system based on HPLC coupled to inductively coupled plasma mass spectrometry to measure the 1,2-GG cisplatin adducts formed in DNA. The method was developed and validated using calf thymus DNA and two different adenocarcinoma cell lines. The values for the limit of detection (LOD) and the limit of quantitation determined for the 1,2-GG cisplatin adduct were 0.21 and 0.67 fmol per microg DNA, respectively. This corresponds to an absolute LOD of 0.8 pg as Pt for the 1,2-GG adduct. Cisplatin-sensitive (H23) and -resistant (A549) tumor cells were exposed to the drug, and the 1,2-GG adduct levels were measured over a 24 h time period. The results showed a statistically significant (P < 0.05) higher concentration in the sensitive cells as compared to the resistant cells after repair for 7 h. Although the adduct concentration present fell at subsequent time points (12 and 24 h), the levels in each cell line were broadly similar. The protocol was then applied to the analysis of patient samples taken before and then 1 h after treatment. The 1,2-GG cisplatin adduct was present in the range from 113 to 1245 fg Pt per microg DNA in all of the patient samples taken after treatment. Although the adduct was not present at levels greater than the LOD in the initial pretreatment samples, trace amounts were discernible in some patient samples on their third treatment cycle.
A novel circular dichroism (CD) method to probe 1-cysteine based on the self-assembly of chiral complex nanoparticles from Ag1 and L-cysteine was reported. To speed up the reaction between Ag1 and L-cysteine, the mixture of Ag1 and L-cysteine was ultrasonicated and heated in a water bath. As a result, the time for the reaction to reach equilibrium was reduced from 100 h to 30 mm because of the synergistic effects of temperature and ultrasonication. To confirm that the CD spectra, after treatment of L-cysteine with Ag1, originated from the differential absorption of left- and right-circularly polarized light rather than the effect of light scattering on small particles in solution, L-cysteine was replaced with D-cysteine. The results show that replacement of L-cysteine with D-cysteine produced an opposite CD signal, and demonstrate that the observed behavior did result from the differential absorption of left- and right circularly polarized light.
Parthenolide is one of the main components responsible for the anti-inflammatory property of Feverfew. Recently, parthenolide has shown to induce apoptosis in cancer cells. Here we further studied the mechanism of parthenolide-induced apoptosis by focusing on the role of intracellular thiols and calcium in a human colorectal cancer cell, COLO 205. Parthenolide rapidly depleted intracellular thiols, including both free glutathione (GSH) and protein thiols. Concomitantly, there were dose- and time-dependent increases in intracellular reactive oxygen species (ROS) and calcium levels. Increased expression of GRP78 protein, a marker for endoplasmic reticulum stress was also detected. All these changes preceded parthenolide-induced apoptotic cell death. More importantly, pretreatment with N-acetylcysteine, a precursor of GSH synthesis, protected the cells from parthenolide-induced thiol depletion, ROS production, cytosolic calcium increase and completely blocked parthenolide-induced apoptosis. On the contrary, pretreatment of buthionine sulfoximine, an inhibitor of GSH synthesis sensitized the cell to apoptosis. These data clearly demonstrate that the intracellular thiols and calcium equilibrium play a critical role in parthenolide-induced apoptotic cell death.
The aggregation-induced emission (AIE) properties of 1,1,2,3,4,5-hexaphenylsilole (HPS) and poly{11-[(1,2,3,4,5-pentaphenylsilolyl)oxy]-1-phenyl-1-undecyne} (PS9PA) were studied by time-resolved fluorescence technique. The enhanced fluorescence and long fluorescent lifetime were obtained for the sample in an aggregate state as compared to the sample in solution. The time-decay of fluorescence of HPS and PS9PA in high viscosity solvents and low-temperature glasses has also been measured in detail to further investigate the possible mechanism for AIE. Enhanced light emission and long fluorescence lifetime were detected for both HPS and PS9PA in the solution-thickening and -cooling experiments. These results provided direct evidence that the enhanced photoluminescence (PL) efficiency is due to restricted intramolecular motion, which ascribes AIE to the deactivation of nonradiative decay caused by restricted torsional motions of the molecules in the solid state or aggregate form.