An ER-Targeting Iridium(III) Complex That Induces Immunogenic Cell Death in Non-Small-Cell Lung Cancer.

Immunogenic cell death (ICD) is a vital component of therapeutically induced anti-tumor immunity. An iridium(III) complex (Ir1), containing an N,N-bis(2-chloroethyl)-azane derivate, as an endoplasmic reticulum-localized ICD inducer for non-small cell lung cancer (NSCLC) is reported. The characteristic discharge of damage-associated molecular patterns (DAMPs), that is, cell surface exposure of calreticulin (CRT), extracellular exclusion of high mobility group box 1 (HMGB1), and ATP, were generated by Ir1 in A549 lung cancer cells, accompanied by an increase in endoplasmic reticulum stress and reactive oxygen species (ROS). The vaccination of immunocompetent mice with Ir1-treated dying cells elicited an antitumor CD8+ T cell response and Foxp3+ T cell depletion, which eventually resulted in long-acting anti-tumor immunity by the activation of ICD in lung cancer cells. Ir1 is the first Ir-based complex that is capable of developing an immunomodulatory response by immunogenic cell death.

FerriIridium: A Lysosome-Targeting Iron(III)-Activated Iridium(III) Prodrug for Chemotherapy in Gastric Cancer Cells.

Reported is the FeIII -activated lysosome-targeting prodrug FerriIridium for gastric cancer theranostics. It contains a meta-imino catechol group that can selectively bond to, and be oxidized by, free FeIII inside the cell. Subsequent oxidative rearrangement releases FeII and hydrolyses the amine bond under acidic conditions, forming an aminobipyridyl Ir complex and 2-hydroxybenzoquinone. Thus, FeII catalyzes the Fenton reaction, transforming hydrogen peroxide into hydroxyl radicals, the benzoquinone compounds interfere with the respiratory chain, and conversion of the prodrug into the Ir complex leads to an increase in phosphorescence and toxicity. These properties, combined with the high FeIII content and acidity of cancer cells, make FerriIridium a selective and efficient theranostic agent (IC50 =9.22 µm for AGS cells vs. >200 µm for LO2 cells). FerriIridium is the first metal-based compound that has been developed for chemotherapy using FeIII to enhance both selectivity and potency.

Bimodal Visualization of Endogenous Nitric Oxide in Lysosomes with a Two-Photon Iridium(III) Phosphorescent Probe.

Nitric oxide (NO) is a fundamental signaling molecule that shows complex effects on the catabolic autophagy process, which is closely linked with lysosomal function. In this study, a new lysosome-targeted, pH-independent, and two-photon phosphorescent iridium(III) complex, Ir-BPDA, has been investigated for endogenous NO detection and imaging. The rational design of the probe, as the addition of the morpholine moieties and the substitution of a benzyl group in the amino group in Ir-BPDA, facilitates its accumulation in lysosomes and makes the reaction product with NO, Ir-BPDA-NO, insusceptible in its phosphorescence intensity and lifetime against pH changes (pH 4-10), well suited for lysosomal NO detection (pH 4-6). Furthermore, Ir-BPDA exhibits a fast and 50-fold response to NO in phosphorescence intensity and a two-photon cross-section as high as 60 GM after the reaction, as well as a notably increased phosphorescence lifetime from 200.1 to 619.6 ns. Thus, accompanied by its photostability, Ir-BPDA enabled the detection of NO in the lipopolysaccharide-stimulated macrophages and zebrafish model, revealing the endogenous lysosomal NO distribution during inflammation in vivo by means of both TPM and PLIM imaging techniques.

Super-Resolution Tracking of Mitochondrial Dynamics with An Iridium(III) Luminophore.

Combining luminescent transition metal complex with super-resolution microscopy is an excellent strategy for the long-term visualization of the dynamics of subcellular structures in living cells. However, it remains unclear whether iridium(III) complexes are applicable for a particular type of super-resolution technique, structured illumination microscopy (SIM), to image subcellular structures. Herein, an iridium(III) dye, to track mitochondrial dynamics in living cells under SIM is described. The dye demonstrates excellent specificity and photostability and satisfactory cell permeability. While using SIM to image mitochondria, an ≈80 nm resolution is achieved that allows the clear observation of the structure of mitochondrial cristae. The dye is used to monitor and quantify mitochondrial dynamics relative to lysosomes, including fusion involved in mitophagy, and newly discovered mitochondria-lysosome contact (MLC) under different conditions. The MLC remains intact and fusion vanishes when five receptors, p62, NDP52, OPTN, NBR1, and TAX1BP1, are knocked out, suggesting that these two processes are independent.

Iridium(III) Anthraquinone Complexes as Two-Photon Phosphorescence Probes for Mitochondria Imaging and Tracking under Hypoxia.

In the present study, four mitochondria-specific and two-photon phosphorescence iridium(III) complexes, Ir1-Ir4, were developed for mitochondria imaging in hypoxic tumor cells. The iridium(III) complex has two anthraquinone groups that are hypoxia-sensitive moieties. The phosphorescence of the iridium(III) complex was quenched by the functions of the intramolecular quinone unit, and it was restored through two-electron bioreduction under hypoxia. When the probes were reduced by reductase to hydroquinone derivative products under hypoxia, a significant enhancement in phosphorescence intensity was observed under one- (λ=405 nm) and two-photon (λ=720 nm) excitation, with a two-photon absorption cross section of 76-153 GM at λ=720 nm. More importantly, these probes possessed excellent specificity for mitochondria, which allowed imaging and tracking of the mitochondrial morphological changes in a hypoxic environment over a long period of time. Moreover, the probes can visualize hypoxic mitochondria in 3D multicellular spheroids and living zebrafish through two-photon phosphorescence imaging.

Cyclometalated Iridium(III) Complexes as Two-Photon Phosphorescent Probes for Specific Mitochondrial Dynamics Tracking in Living Cells.

Five cyclometalated iridium(III) complexes with 2-phenylimidazo[4,5-f][1,10]phenanthroline derivatives (IrL1-IrL5) were synthesized and developed to image and track mitochondria in living cells under two-photon (750 nm) excitation, with two-photon absorption cross-sections of 48.8-65.5 GM at 750 nm. Confocal microscopy and inductive coupled plasma-mass spectrometry (ICP-MS) demonstrated that these complexes selectively accumulate in mitochondria within 5 min, without needing additional reagents for membrane permeabilization, or replacement of the culture medium. In addition, photobleaching experiments and luminescence measurements confirmed the photostability of these complexes under continuous laser irradiation and physiological pH resistance. Moreover, results using 3D multicellular spheroids demonstrate the proficiency of these two-photon luminescent complexes in deep penetration imaging. Two-photon excitation using such novel complexes of iridium(III) for exclusive visualization of mitochondria in living cells may substantially enhance practical applications of bioimaging and tracking.

Metal-Free Carbon Nitride Nanosheet Supported the Pentacoordinated Silicon Intermediates for Photocatalytic Overall Water Splitting.

Photocatalytic overall water splitting is a promising approach to overcome the environmental and energy crisis. However, developing effective photocatalysts with well activity, light absorption, and photogenerated carrier lifetime is still a challenge. Herein, combining extensive first-principles and nonadiabatic molecular dynamics calculations, we find that microporous carbon-nitride nanosheets with a pyridinic nitrogen, such as C2N and C6N6, possess the pentacoordinated silicon intermediates' bonding environment. The pentacoordinated silicon as intermediates exhibits good photocatalytic performance for the difficult four-electronic oxygen evolution reaction. The overpotential is only 0.55 V for C2N, which is significantly lower than that of the tetracoordinated silicon intermediates (2.00 V). Simultaneously, the decoration of the silicon group not only widens the absorption range of visible light but also maintains the lifetime of photogenerated carriers on the nanosecond scale, which enhances the application efficiency of solar energy. Our work paves a new route for advancing photocatalytic overall water splitting.

Cyclometalated Iridium(III) Complexes as Mitochondria-targeting Photosensitizers against Cisplatin-resistant Cells.

Four iridium (III) complexes Ir1-Ir4 were synthesized and characterized. Possessing high singlet oxygen production ability and specific mitochondria-localization, Ir1 was developed as a mitochondria-targeting photosensitizer. Ir1 exhibited strong phototoxicity against cancer cell line A549 and its corresponding cisplatin-resistant one A549R. In contrast, Ir1 showed low cytotoxicity toward normal cell HLF. This selectivity resulted from the different uptake amount. With 405 nm irradiation, Ir1 induced mitochondria-mediated cell death in A549R cells, achieving the overcome of drug-resistant.

Anti-metastasis and anti-proliferation effect of mitochondria-accumulating ruthenium(II) complexes via redox homeostasis disturbance and energy depletion.

The antiproliferative activity of three cyclometalated Ru(II) complexes with the formula [Ru(bpy)2L]PF6, where bpy = 2,2'-bipyridine, Ru1: L1 = phenanthro[4,5-fgh]quinoxaline; Ru2: L2 = benzo[f]naphtho[2,1-h]quinoxaline; and Ru3: L3 = phenanthro[9,10-b]pyrazine, have been synthesized and characterized. The lipophilicity of the three Ru(II) complexes was modulated by the alteration of the planarity in the ligands of the complexes. With appropriate lipophilicity, Ru1-Ru3 exhibited mitochondrial accumulating property and cytotoxic activity against a spectrum of cancer cell lines. The underlying mechanism study indicated that these Ru(II) complexes can selectively accumulate in mitochondria and disrupt physiological processes, including the redox balance and energy generation in cancer cells. Elevation of iron content in triple-negative breast cancer (MDA-MB-231 cells) was observed after treatment with Ru(II) complexes, which may contribute to the production of reactive oxygen species (ROS) via Fenton reaction chemistry. Besides, the Ru(II) complexes decreased the intracellular glutathione (GSH) in cancer cells, leading to the failure in the cells to combat oxidative damage. Both of the mentioned processes contribute to the high oxidative stress and eventually lead to cancer cell death. On the other hand, Ru1-Ru3 significantly induced the depletion of adenosine triphosphate (ATP), causing disturbance of energy generation. Moreover, the results of wound-healing assay and transwell invasion assay, as well as the tube formation assay indicated the anti-migration and anti-angiogenesis properties of Ru1-Ru3. Our study demonstrated that these Ru(II) complexes are promising chemotherapeutic agents with oxidative stress induction and energy generation disturbance.

Iridium(iii) complexes as mitochondrial topoisomerase inhibitors against cisplatin-resistant cancer cells.

Herein, we developed the first metal-based mitochondrial topoisomerase inhibitors to achieve an effective therapeutic outcome for the therapy of cisplatin-resistant tumour cells.

Metal complexes for mitochondrial bioimaging.

Mitochondria are essential organelles in eukaryotic cells, containing various signaling molecules and important enzymes associated with cell growth, death, and proliferation. The visualization of mitochondria and their biochemistry with confocal microscopy, fluorescence (phosphorescence) lifetime microscopy (FLIM, PLIM), and super-resolution microscopy has therefore been of great interest in recent years. In particular, transition metal complexes have emerged as excellent mitochondria-targeting probes, due to their high photostabilities, large Stokes shifts, tunable chemical structures and long luminescence lifetimes. In this review, we focus on platinum, ruthenium and iridium complexes, and their application as detectors of micro-environmental alterations as well as for the imaging of signaling molecules inside mitochondria.

Quantitative analysis of interactive behavior of mitochondria and lysosomes using structured illumination microscopy.

Super-resolution optical microscopy has extended the spatial resolution of cell biology from the cellular level to the nanoscale, enabling the observation of the interactive behavior of single mitochondria and lysosomes. Quantitative parametrization of interactions between mitochondria and lysosomes under super-resolution optical microscopy, however, is currently unavailable, which has severely limited our understanding of the molecular machinery underlying mitochondrial functionality. Here, we introduce an M-value to quantitatively investigate mitochondria and lysosome contact (MLC) and mitophagy under structured illumination microscopy. We found that the M-value for an MLC is typically less than 0.4, whereas in mitophagy it ranges from 0.5 to 1.0. This system permits further investigation of the detailed molecular mechanism governing the interactive behavior of mitochondria and lysosomes.

Endoplasmic reticulum targeted cyclometalated iridium(iii) complexes as efficient photodynamic therapy photosensitizers.

The endoplasmic reticulum (ER) is an indispensable organelle that undertakes the synthesis and export of proteins and membrane lipids. Subtle interferences of the ER redox signaling pathway are very likely to cause ER-stress induced apoptosis. In view of this, we herein present a series of ER-targeted Ir(iii) complexes (Ir1-Ir3) as photodynamic therapy (PDT) photosensitizers with a gradually extended conjugation area in the main ligand, and study the correlation between the conjugation area and PDT performance. The results showed that all of these complexes can accumulate in the ER and effectively induce cell apoptosis after PDT therapeutics (405 nm, 6 J cm-2) by an ER stress mechanism, and both their singlet oxygen quantum yields and cytotoxicities increase as the conjugation area extends. All complexes showed PDT efficacy towards different cancer cell lines. Among them, Ir2 exhibited the highest PI value (94.3) against A549 cells with an IC50 down to 0.65 µM. In addition, the post PDT ER-stress induced apoptosis along with the efflux of Ca2+ from the ER system in A549 cells in a short period of time (45-90 min) with the pretreatment of Ir2 was demonstrated. All of these results indicate the promising potential of Ir2 as an effective PDT photosensitizer.

A mitochondria-targeting dinuclear Ir-Ru complex as a synergistic photoactivated chemotherapy and photodynamic therapy agent against cisplatin-resistant tumour cells.

A mitochondria-targeting hetero-binuclear Ir(iii)-Ru(ii) complex was developed as a photoactivated chemotherapy (PACT) and photodynamic therapy (PDT) bifunctional agent to achieve a synergistic effective therapeutic outcome for the therapy of cisplatin-resistant tumour cells.

Oncosis-inducing cyclometalated iridium(iii) complexes.

Oncosis is a non-apoptotic form of programmed cell death (PCD), which differs from apoptosis in both morphological changes and inner pathways, and might hold the key to defeating a major obstacle in cancer therapy - drug-resistance, which is often a result of the intrinsic apoptosis resistance of tumours. However, despite the fact that the term "oncosis" was coined and used much earlier than apoptosis, little effort has been made to discover new drugs which can initiate this form of cell death, in comparison to drugs inducing apoptosis or any other type of PCD. So herein, we present the synthesis of a series of mitochondria-targeting cyclometalated Ir(iii) complexes, which activated the oncosis-specific protein porimin and calpain in cisplatin-resistant cell line A549R, and determined their cytotoxicity against a wide range of drug-resistant cancer types. To the best of our knowledge, these complexes are the very first metallo-components to induce oncosis in drug-resistant cancer cells.

Rational design of NIR-emitting iridium(iii) complexes for multimodal phosphorescence imaging of mitochondria under two-photon excitation.

A series of NIR-emitting iridium(iii) complexes were developed for multimodal phosphorescence imaging (NIR imaging, phosphorescence lifetime imaging and time-gated imaging) of mitochondria in living cells, 3D multicellular spheroids (MTCCs) and hippocampus slice under two-photon excitation.

Ruthenium(II) Complexes with 2-Phenylimidazo[4,5-f][1,10]phenanthroline Derivatives that Strongly Combat Cisplatin-Resistant Tumor Cells.

Cisplatin was the first metal-based therapeutic agent approved for the treatment of human cancers, but its clinical activity is greatly limited by tumor drug resistance. This work utilized the parent complex [Ru(phen)2(PIP)](2+) (1) to develop three Ru(II) complexes (2-4) with different positional modifications. These compounds exhibited similar or superior cytotoxicities compared to cisplatin in HeLa, A549 and multidrug-resistant (A549R) tumor cell lines. Complex 4, the most potent member of the series, was highly active against A549R cancer cells (IC50 = 0.8 µM). This complex exhibited 178-fold better activity than cisplatin (IC50 = 142.5 µM) in A549R cells. 3D multicellular A549R tumor spheroids were also used to confirm the high proliferative and cytotoxic activity of complex 4. Complex 4 had the greatest cellular uptake and had a tendency to accumulate in the mitochondria of A549R cells. Further mechanistic studies showed that complex 4 induced A549R cell apoptosis via inhibition of thioredoxin reductase (TrxR), elevated intracellular ROS levels, mitochondrial dysfunction and cell cycle arrest, making it an outstanding candidate for overcoming cisplatin resistance.

Cyclometalated Iridium(III) Complexes as AIE Phosphorescent Probes for Real-Time Monitoring of Mitophagy in Living Cells.

Mitophagy, which is a special autophagy that removes damaging mitochondria to maintain sufficient healthy mitochondria, provides an alternative path for addressing dysfunctional mitochondria and avoiding cellular death. In the present study, by coupling the triphenylamine group with 2-phenylimidazo[4,5-f][1,10]phenanthroline derivatives, we synthesized five Ir(III) complexes with an AIE property that are expected to fulfill requirements for real-time monitoring of mitophagy. Ir1-Ir5 were exploited to image mitochondria with a short incubation time by confocal microscopy and inductive coupled plasma-mass spectrometry (ICP-MS). Due to aggregation-induced emission (AIE), Ir1-Ir5 exhibited excellent photostability compared to MitoTracker Green (MTG). Moreover, Ir1-Ir5 manifested satisfactory photostability in the mitochondrial physiological pH range. In addition, the uptake mechanism of Ir1 was investigated using confocal microscopy and flow cytometry analysis. Finally, using both Ir1 and LysoTracker Green, we were able to achieve real-time monitoring of mitophagy.

Cyclometalated iridium(III) complexes as mitochondria-targeted anticancer agents.

Four cyclometalated iridium(III) complexes [Ir(dfppy)2(L)](+) (dfppy = 2-(2,4-difluorophenyl)pyridine, L = 6-(pyridin-2-yl)-1,3,5-triazine-2,4-diamine, Ir1; 6-(isoquinolin-1-yl)-1,3,5-triazine-2,4-diamine, Ir2; 6-(quinolin-2-yl)-1,3,5-triazine-2,4-diamine, Ir3; 6-(isoquinolin-3-yl)-1,3,5-triazine-2,4-diamine, Ir4) have been synthesized and characterized. Distinct from cisplatin, Ir1-Ir4 could specifically target mitochondria and induced apoptosis against various cancer cell lines, especially for cisplatin resistant cells. ICP-MS results indicated that Ir1-Ir4 were taken up via different mechanism for cancer cells and normal cells, which resulted in their high selectivity. The structure-activity relationship and signaling pathways were also discussed.

Cyclometalated iridium(III) complexes with imidazo[4,5-f][1,10]phenanthroline derivatives for mitochondrial imaging in living cells.

A new series of cyclometalated iridium(III) complexes with imidazo[4,5-f][1,10]phenanthroline derivatives (i.e., MitoIr1-MitoIr7) were synthesized and developed to image mitochondria in living cells. In comparison with commercially available mitochondrial trackers, these complexes exhibit a superior capacity to selectively accumulate in mitochondria with no requirement of any membrane permeabilization or replacement of the culture medium. In addition, the excellent photostability under continuous laser irradiation as well as the stable physiological pH resistance of these complexes were confirmed by photobleaching experiments and luminescence measurements. Importantly, MitoIr7, which exhibited both excellent luminescence and high ability to locate in mitochondria, was developed to track the mitochondrial morphological changes over a long period of time.