Triplet-Triplet Energy Transfer in Bis-Cyclometalated Iridium Complexes with Pyrene-Substituted Isocyanides.

Nonlinear optical (NLO) materials are able to modulate responses of electromagnetic radiation, leading to phenomena critical to modern telecommunications technologies. The last two decades have seen significant advances in the area of molecular nonlinear chromophores, particularly with respect to reverse-saturable absorption (RSA). Here, we introduce a strategy for intense excited-state absorption (ESA) that involves bis-cyclometalated iridium complexes with isocyanide ancillary ligands decorated with pyrene triplet acceptors. Upon excitation, the complexes undergo rapid triplet-triplet energy transfer (TTET) to the acceptor excited states. This report describes five bis-cyclometalated iridium complexes using two different pyrene-substituted isocyanides with the general formula [Ir(C^N)2(CNAr)2]PF6 (C^N = cyclometalating ligand, CNAr = isocyanide ancillary ligand: CNArpyr = 2,6-dimethyl-4-(1-pyrenyl)phenyl isocyanide, CNpyr = 1-pyrenyl isocyanide). The synthesized complexes were thoroughly characterized via 1H and 13C{1H} NMR spectroscopy, Fourier-transform Infrared spectroscopy, and electrospray ionization mass spectrometry. The excited states were evaluated with UV-vis absorption, steady-state and time-resolved photoluminescence, and transient absorption spectroscopy. Phosphorescence is completely quenched at room temperature, but in the solvent glass matrix at 77 K, there is luminescence originating from a π → π* triplet state on the pyrene moiety, abbreviated herein as 3pyrene. All five complexes display intense and long-lived ESA originating from the 3pyrene state. The localization of the ground-state absorption on the cyclometalating ligands and the excited-state absorption on the pyrene moiety allows for independent tuning of ground-state absorption (GSA) and ESA to optimize RSA and other NLO attributes.

Panchromatic Excited-State Absorption in Bis-Cyclometalated Iridium Isocyanide Complexes.

In this work, we introduce a series of cyclometalated iridium complexes and evaluate the suitability of this class of compounds in nonlinear optical (NLO) applications, with an emphasis on long-lived, panchromatic reverse-saturable absorption (RSA). The investigated complexes are represented by the general formula [Ir(C^N)2(CNArdmp)2]+, (C^N = cyclometalating ligand, CNArdmp = 2,6-dimethylphenyl isocyanide). Seven such complexes were synthesized and characterized, including in-depth analysis of their photophysical properties (UV-vis absorption, photoluminescence, and transient absorption). This series of compounds contains seven different cyclometalating ligands (2-phenylbenzothiazole (bt) (Ir1), 5-nitro-2-phenylpyridine (ppyNO2) (Ir2), 5-nitro-2-(9-phenanthryl)pyridine (phenNO2) (Ir3), 2-(benzo[b]thiophen-2-yl)quinoline (btq) (Ir4), 6-(benzo[b]thiophen-2'-yl)phenanthridine (btph) (Ir5), 2,4-diphenylquinoline (dpq) (Ir6), and 6-nitro-2,4-diphenylquinoline (NO2dpq) (Ir7)), which have profound effects on their ground-state and excited-state absorption spectra. To evaluate the effects of the isocyanide ancillary ligands, some heteroleptic bis-cyclometalated iridium(III) acetylacetonate (acac) analogue complexes are included as points of comparison. In the ground state, the bis-isocyanide complexes display UV-vis absorption with the characteristic 1LC (π → π*) band at λ < 350 nm and 1MLCT bands at 350-500 nm. Five of the complexes (Ir1, Ir2, Ir4, Ir5, and Ir6) display broad, intense triplet excited-state absorption with no ground-state bleach (GSB) over the spectral window of 400-900 nm, with excited-state lifetimes spanning three orders of magnitude from ∼32 ns to 12 µs. The photophysical data suggests that the isocyanide ancillary ligand blue-shifts the GSB transient into the UV, which is normally found in the middle of the visible region for cyclometalated iridium complexes. This study demonstrates the applicability of cationic cyclometalated iridium(III) bis-isocyanide complexes as candidate RSA materials.

Ru(II)-Based Acetylacetonate Complexes Induce Apoptosis Selectively in Cancer Cells.

The synthesis, chemical and biological characterization of seven Ru(II) polypyridyl complexes containing acetylacetonate (acac) ligands are reported. Electronic absorption spectra were determined and electrochemical potentials consistent with Ru(III/II) couples ranging from +0.60 to +0.73 V vs Ag/AgCl were measured. A series of complexes were screened against MDA-MB-231, DU-145, and MCF-10A cell lines to evaluate their cytotoxicities in cancer and normal cell lines. Although most complexes were either nontoxic or equipotent in cancer cells and normal cell lines, compound 1, [Ru(dpqy)(acac)(py)](PF6), where dqpy is 2,6-di(quinolin-2-yl)pyridine, showed up to 2.5:1.0 selectivity for cancer as compared to normal cells, along with nanomolar EC50 values in MDA-MB-231 cells. Lipophilicity, determined as the octanol/water partition coefficient, log Po/w, ranged from -0.33 (0.06) to 1.15 (0.10) for the complexes. Although cytotoxicity was not correlated with electrochemical potentials, a moderate linear correlation between lipophilicity and toxicities was observed. Cell death mechanism studies indicated that several of the Ru-acac compounds, including 1, induce apoptosis in MDA-MB-231 cells.

ARL Spectral Fitting as an Application to Augment Spectral Data via Franck-Condon Lineshape Analysis and Color Analysis.

The ARL Spectral Fitting application provides a free, publicly accessible, and fully transparent method for performing Franck-Condon Lineshape Analysis (FCLSA) on spectral data, in addition to CIE color coordinate determination and basic spectral processing. While some of the features may be found in commercial software or in programs made by academic research groups, we believe that ARL Spectral Fitting is the only application that possesses all three of the aforementioned attributes. This program is intended as a standalone, GUI-based application for use by an average laboratory researcher without requiring any coding knowledge or proprietary software. In addition to the standalone executable hosted on ARL GitHub, the associated MATLAB files are available for use and further development. FCLSA augments the information found in luminescence spectra, providing meaningful insight into the relationship between the ground and excited states of a photoluminescent species. This insight is achieved by modeling spectra with two versions (modes) of an equation that are characterized by either four or six parameters, depending on which mode is used. Once optimized, the value of each of these parameters can be used to gain insight into the molecule, as well as to perform further analysis (for example, the free energy content of the excited-state molecule). This application provides tools for easy by-hand fitting of imported data, as well as two methods for optimizing this fit-damped least-squares fitting, powered by the Levenberg-Marquardt algorithm, and derivative-free fitting utilizing the Nelder-Mead simplex algorithm. Furthermore, estimations of sample color can be performed and reported in CIE and RGB coordinates.

Photosensitive Ru(II) Complexes as Inhibitors of the Major Human Drug Metabolizing Enzyme CYP3A4.

We report the synthesis and photochemical and biological characterization of the first selective and potent metal-based inhibitors of cytochrome P450 3A4 (CYP3A4), the major human drug metabolizing enzyme. Five Ru(II)-based derivatives were prepared from two analogs of the CYP3A4 inhibitor ritonavir, 4 and 6: [Ru(tpy)(L)(6)]Cl2 (tpy = 2,2':6',2″-terpyridine) with L = 6,6'-dimethyl-2,2'-bipyridine (Me2bpy; 8), dimethylbenzo[i]dipyrido[3,2-a:2',3'-c]phenazine (Me2dppn; 10) and 3,6-dimethyl-10,15-diphenylbenzo[i]dipyrido[3,2-a:2',3'-c]phenazine (Me2Ph2dppn; 11), [Ru(tpy)(Me2bpy)(4)]Cl2 (7) and [Ru(tpy)(Me2dppn)(4)]Cl2 (9). Photochemical release of 4 or 6 from 7-11 was demonstrated, and the spectrophotometric evaluation of 7 showed that it behaves similarly to free 4 (type II heme ligation) after irradiation with visible light but not in the dark. Unexpectedly, the intact Ru(II) complexes 7 and 8 were found to inhibit CYP3A4 potently and specifically through direct binding to the active site without heme ligation. Caged inhibitors 9-11 showed dual action properties by combining photoactivated dissociation of 4 or 6 with efficient 1O2 production. In prostate adenocarcinoma DU-145 cells, compound 9 had the best synergistic effect with vinblastine, the anticancer drug primarily metabolized by CYP3A4 in vivo. Thus, our study establishes a new paradigm in CYP inhibition using metalated complexes and suggests possible utilization of photoactive CYP3A4 inhibitory compounds in clinical applications, such as enhancement of therapeutic efficacy of anticancer drugs.

Anticancer and antitrypanosomal activities of trinuclear ruthenium compounds with orthometalated phenazine ligands.

Trinuclear ruthenium complexes with orthometalated phenazines of general formula [Ru3(µ3-O)(µ2-OAc)5(L)(py)2]PF6 (L = dppn, benzo[i]dipyrido[3,2-a:2',3'-c]phenazine, 1; dppz, dipyrido[3,2-a:2',3'-c]phenazine, 2; CH3-dppz, 7-methyldipyrido[3,2-a:2',3'-c]phenazine, 3; Cl-dppz, 7-chlorodipyrido[3,2-a:2',3'-c]phenazine, 4) were investigated for their cytotoxic activity toward the B16F10 murine melanoma and the L929 non-cancer cell lines and against Trypanosoma cruzi (2-4). This study also reports a multi-technique investigation into how complexes 1-4 interact with DNA and human serum albumin, HSA. At concentrations ranging from 2 to 50 µM, all the complexes reduced B16F10 murine melanoma cell viability by over 50%. Complex 4 had the highest cytotoxic effect in the series, diminishing B16F10 cell viability to 38% at 2 µM, with an overall order for anticancer activity of 4 > 2 > 3 > 1. Complexes 2-4 showed remarkable activity in inhibiting epimastigote and amastigote forms of T. cruzi. Complex 2 showed better antitrypanosomal activity than the reference drug (IC50 = 1.19 µM and IC50 = 0.25 µM for epimastigote and amastigotes forms, respectivily). Ethidium bromide (EB) displacement assays showed that DNA intercalation progressively increases with the extension of the π-conjugation of the cyclometalating ligand and the presence of substituents in the phenazinic portion (1 > 4-3 > 2), showing that complex 1 is a stronger intercalator than EB itself (Kapp > 107 M-1). Viscosity measurements followed the same trend. Cytotoxicity against cancer cells and antitrypanosomal activity follow the same order, which is different to the tendency of DNA intercalation, suggesting DNA is not the main target of these complexes. Compound 1-4 showed very high affinity with HSA (Kb ∼109 M-1). Circular dichroism results also showed that the complexes alter significantly the secondary structure of the HSA, lowering the α-helix % from 86.2 (pure protein) to less than 5% for compounds 1, 2 and 4 at 2.8 µM. These findings demonstrated the important role of phenazines for the biological activity of triruthenium compounds.

New Ru(ii) photocages operative with near-IR light: new platform for drug delivery in the PDT window.

A series of Ru(ii) complexes bearing the tridentate 2,6-di(quinolin-2-yl)pyridine (dqpy) ligand were designed to undergo photoinduced ligand dissociation with red/near-IR light. The complexes [Ru(dqpy)(L)(CH3CN)]2+, where L = 2,2'-bipyridine (bpy, 1), 4,4'dimethyl-2,2'-bipyridine (Me2bpy, 2), and 1,10-phenanthroline (phen, 3). Complexes 1-3 exhibit red-shifted lowest energy metal-to-ligand charge transfer (MLCT) absorption maxima at ∼600 nm, as compared to the corresponding tpy (2,2';6',2''-terpyridine) complexes with MLCT bands at ∼565 nm which appear as shoulders to the MLCT bands at ∼455 nm. This shift is attributed to the lower energy LUMO afforded by the dqpy ligand when compared to tpy, as evidenced by the shift of the first reduction wave to ∼0.3 V more positive potentials in the former. In addition, the lowest MLCT maximum of [Ru(dqpy)(acac)(CH3CN)]+ (4; acac- = acetylacetonate) is observed at 770 nm, attributed to the additional increase in energy of the HOMO afforded by the presence of the π-donating acac- ligand and supported by calculations. Complexes 1-3 undergo ligand substitution upon irradiation with red light, λ irr ≥ 610 nm, and the ligand substitution photochemistry of 4 is accessible with near-IR light, λ irr ≥ 715 nm and λ irr = 735 nm. Complexes 1-4 exhibit similar quantum yields of ligand exchange, Φ L, with 450 and 600 nm irradiation, however, that of 4 is 2-3 times greater than those measured for 1-3. This enhancement is explained by the difference in ligand contributions to the HOMO. Density functional theory calculations predict partial dqpy ππ* character in the MLCT states of 1-3 and a mixed Ru/acac- → dqpy metal/ligand-to-ligand charge transfer (ML-LCT) state in 4. The photoreactivity of 1-4 with tissue-penetrating red and near-IR light, together with their exceptional dark stability (>48 h), makes the new Ru(ii)-dqpy platform ideal for the development of new complexes for photoinduced drug release and for other applications that require broad absorption from the ultraviolet and visible ranges into the near-IR, such as solar energy conversion.

Catch and Release Photosensitizers: Combining Dual-Action Ruthenium Complexes with Protease Inactivation for Targeting Invasive Cancers.

Dual action agents containing a cysteine protease inhibitor and Ru-based photosensitizer for photodynamic therapy (PDT) were designed, synthesized, and validated in 2D culture and 3D functional imaging assays of triple-negative human breast cancer (TNBC). These combination agents deliver and release Ru-based PDT agents to tumor cells and cause cancer cell death upon irradiation with visible light, while at the same time inactivating cathespin B (CTSB), a cysteine protease strongly associated with invasive and metastatic behavior. In total five Ru-based complexes were synthesized with the formula [Ru(bpy)2(1)](O2CCF3)2 (3), where bpy = 2,2'-bipyridine and 1 = a bipyridine-based epoxysuccinyl inhibitor; [Ru(tpy)(NN)(2)](PF6)2, where tpy = terpiridine, 2 = a pyridine-based epoxysuccinyl inhibitor and NN = 2,2'-bipyridine (4); 6,6'-dimethyl-2,2'-bipyridine (5); benzo[ i]dipyrido[3,2- a:2',3'- c]phenazine (6); and 3,6-dimethylbenzo[ i]dipyrido[3,2- a:2',3'- c]phenazine (7). Compound 3 contains a [Ru(bpy)3]2+ fluorophore and was designed to track the subcellular localization of the conjugates, whereas compounds 4-7 were designed to undergo either photoactivated ligand dissociation and/or singlet oxygen generation. Photochemical studies confirmed that complexes 5 and 7 undergo photoactivated ligand dissociation, whereas 6 and 7 generate singlet oxygen. Inhibitors 1-7 all potently and irreversibly inhibit CTSB. Compounds 4-7 were evaluated against MDA-MB-231 TNBC and MCF-10A breast epithelial cells in 2D and 3D culture for effects on proteolysis and cell viability under dark and light conditions. Collectively, these data reveal that 4-7 potently inhibit dye-quenched (DQ) collagen degradation, whereas only compound 7 causes efficient cell death under light conditions, consistent with its ability to release a Ru(II)-based photosensitizer and to also generate 1O2.

Photoactivation of imatinib-antibody conjugate using low-energy visible light from Ru(ii)-polypyridyl cages.

Ru(ii)-polypyridyl cages with sterically bulky bidentate ligands provide efficient photochemical release of the anticancer drug imatinib using low energy visible light, imparting spatiotemporal control over drug bioavailability. The light-activated drug release is maintained when the Ru(ii) cage is covalently coupled to an antibody, which is expected to localize selectively on the tumor.

New Ru(ii) complex for dual photochemotherapy: release of cathepsin K inhibitor and 1O2 production.

A new complex, [Ru(tpy)(dppn)(Cbz-Leu-NHCH2CN)]2+ (1, tpy = 2,2':6',2''-terpyridine, dppn = benzo[i]dipyrido[3,2-a:2',3'-c]phenazine) was synthesized and its photochemical properties were investigated. This complex undergoes photorelease of the Cbz-Leu-NHCH2CN ligand, a known cathepsin K inhibitor, with a quantum yield, Φ450, of 0.0012(4) in water (λirr = 450 nm). In addition, 1 sensitizes the production of singlet oxygen upon visible light irradiation with quantum yield, ΦΔ, of 0.64(3) in CH3OH. The photophysical properties of 1 were compared with those of [Ru(tpy)(bpy)(Cbz-Leu-NHCH2CN)]2+ (2, bpy = 2,2'-bipyridine), [Ru(tpy)(dppn)(CH3CN)]2+ (3), and [Ru(tpy)(bpy)(CH3CN)]2+ (4) to evaluate the effect of the release of the Cbz-Leu-NHCH2CN inhibitor relative to the CH3CN ligand, as well as the role of dppn as the bidentate ligand for 1O2 production instead of bpy. Nanosecond transient absorption spectroscopy confirms the formation of the long-lived dppn-centered 3ππ* state in 1 and 3 with a maximum at ∼540 nm and τ ∼20 µs in deaerated acetonitrile. Complexes 1 and 3 are able to cause photoinduced damage to DNA (λirr ≥ 395 nm), whereas 2 and 4 do not photocleave DNA under similar experimental conditions. These results suggest that 1 is a promising agent for dual activity, both releasing a drug and producing singlet oxygen, and is poised to exhibit enhanced biological activity in phototochemotherapy upon irradiation with visible light.