Photoactivated Chemotherapy Research Articles

Partially Solvated Dinuclear Ruthenium Compounds Bridged by Quinoxaline-Functionalized Ligands as Ru(II) Photocage Architectures for Low-Energy Light Absorption.

Ruthenium compounds with coordinated photolabile molecules that can be selectively released by irradiation with a visible light source are finding increasing applications in photoactivated chemotherapy (PCT) as photocages. Earlier photocages based on mononuclear Ru(II) compounds lack absorption in the therapeutic window (λ > 600 nm). In previous work, we synthesized the first partially solvated tppz bridged (tppz= 2,3,5,6-tetrakis(pyridin-2-yl)pyrazine) dinuclear Ru(II) complex capable of photoinduced ligand exchange at both metal centers. To further explore the effect of the bridging ligand on Ru(II) photocage design, we used quinoxaline-functionalized bridging ligand platforms to prepare [{RuII(NCCH3)4}2(μ-BL)](PF6)4[BL = dpq, 2,3-di(pyridin-2-yl)quinoxaline (1); BL = dpb, 2,3-di(pyridin-2-yl)benzo[g]quinoxaline (2)]. The compounds are capable of absorbing green light with tails extending beyond 650 nm which can be exploited for applications as PCT agents. Experimental results were additionally verified by DFT calculations. The use of two Ru(II) centers equipped with quinoxaline-based bridging ligands is a promising design strategy for the synthesis of a new family of dinuclear Ru(II) photocage prototypes with the ability to absorb low-energy visible light.

Photo-Uncaging of a Microtubule-Targeted Rigidin Analogue in Hypoxic Cancer Cells and in a Xenograft Mouse Model.

Marine alkaloid rigidins are cytotoxic compounds known to kill cancer cells at nanomolar concentrations by targeting the microtubule network. Here, a rigidin analogue containing a thioether group was "caged" by coordination of its thioether group to a photosensitive ruthenium complex. In the dark, the coordinated ruthenium fragment prevented the rigidin analogue from inhibiting tubulin polymerization and reduced its toxicity in 2D cancer cell line monolayers, 3D lung cancer tumor spheroids (A549), and a lung cancer tumor xenograft (A549) in nude mice. Photochemical activation of the prodrug upon green light irradiation led to the photosubstitution of the thioether ligand by water, thereby releasing the free rigidin analogue capable of inhibiting the polymerization of tubulin. In cancer cells, such photorelease was accompanied by a drastic reduction of cell growth, not only when the cells were grown in normoxia (21% O2) but also remarkably in hypoxic conditions (1% O2). In vivo, low toxicity was observed at a dose of 1 mg·kg-1 when the compound was injected intraperitoneally, and light activation of the compound in the tumor led to 30% tumor volume reduction, which represents the first demonstration of the safety and efficacy of ruthenium-based photoactivated chemotherapy compounds in a tumor xenograft.

Towards Light-Activated Ruthenium-Arene (RAPTA-Type) Prodrug Candidates.

Cancer is currently one of the deadliest diseases worldwide. Based on the high incidence of this disease, the side effects associated with current chemotherapies and the appearance of drug resistance, considerable efforts have been directed towards the development of new anticancer drugs with new modes of action. Metal-based compounds are particularly attractive candidates due to their metabolic mechanisms, which differ substantially from those of organic drugs. Of special interest in this context are organometallic ruthenium(II) complexes of the type [Ru(η6 -arene)(pta)Cl2 ] (arene: p-cymene, toluene, benzene, etc.; pta: 1,3,5-triaza-7-phosphaadamantane), which are abbreviated to RAPTA. Complementary to chemotherapy, photoactivated chemotherapy is a technique that has received increasing attention towards the development of treatment for numerous kinds of cancer. With this in mind, a photoactive RAPTA-type complex bearing azide ligands has been designed. The diazide complex, [Ru(η6 -p-cymene)pta-(N3 )2 ], is inert in water, but slowly releases the azide ligand upon exposure to light. Consequently, the in vitro cytotoxicity of the complex in the dark and upon light exposure at λ=450 nm in human cervical carcinoma (HeLa) and noncancerous retinal pigment epithelium (RPE-1) cells was investigated. Although the cytotoxicity of the complex was found to be modest in the dark, an increase in toxicity upon light exposure was observed.

New Designs for Phototherapeutic Transition Metal Complexes

AbstractIn this Minireview, we highlight recent advances in the design of transition metal complexes for photodynamic therapy (PDT) and photoactivated chemotherapy (PACT), and discuss the challenges and opportunities for the translation of such agents into clinical use. New designs for light‐activated transition metal complexes offer photoactivatable prodrugs with novel targeted mechanisms of action. Light irradiation can provide spatial and temporal control of drug activation, increasing selectivity and reducing side‐effects. The photophysical and photochemical properties of transition metal complexes can be controlled by the appropriate choice of the metal, its oxidation state, the number and types of ligands, and the coordination geometry.

New Designs for Phototherapeutic Transition Metal Complexes.

In this Minireview, we highlight recent advances in the design of transition metal complexes for photodynamic therapy (PDT) and photoactivated chemotherapy (PACT), and discuss the challenges and opportunities for the translation of such agents into clinical use. New designs for light‐activated transition metal complexes offer photoactivatable prodrugs with novel targeted mechanisms of action. Light irradiation can provide spatial and temporal control of drug activation, increasing selectivity and reducing side‐effects. The photophysical and photochemical properties of transition metal complexes can be controlled by the appropriate choice of the metal, its oxidation state, the number and types of ligands, and the coordination geometry.

Diastereoselective Synthesis and Two-Step Photocleavage of Ruthenium Polypyridyl Complexes Bearing a Bis(thioether) Ligand.

Thioethers are good ligands for photoactivatable ruthenium(II) polypyridyl complexes, as they form thermally stable complexes that are prone to ligand photosubstitution. Here, we introduce a novel symmetric chelating bis(thioether) ligand scaffold, based on 1,3-bis(methylthio)-2-propanol (4) and report the synthesis and stereochemical characterization of the series of novel ruthenium(II) polypyridyl complexes [Ru(bpy)2(L)](PF6)2 ([1]–[3](PF6)2), where L is ligand 4, its methyl ether, 1,3-bis(methylthio)-2-methoxypropane (5), or its carboxymethyl ether, 1,3-bis(methylthio)-2-(carboxymethoxy)propane (6). Coordination of ligands 4–6 to the bis(bipyridine)ruthenium center gives rise to 16 possible isomers, consisting of 8 possible Λ diastereoisomers and their Δ enantiomers. We found that the synthesis of [1]–[3](PF6)2 is diastereoselective, yielding a racemic mixture of the Λ-(S)-eq-(S)-ax-OHeq-[Ru]2+ and Δ-(R)-ax-(R)-eq-OHeq-[Ru]2+ isomers. Upon irradiation with blue light in water, [1]–[3](PF6)2 selectively substitute their bis(thioether) ligands for water molecules in a two-step photoreaction, ultimately producing [Ru(bpy)2(H2O)2]2+ as the photoproduct. The relatively stable photochemical intermediate was identified as cis-[Ru(bpy)2(κ1-L)(H2O)]2+ by mass spectrometry. Global fitting of the time evolution of the UV–vis absorption spectra of [1]–[3](PF6)2 was employed to derive the photosubstitution quantum yields (Φ443) for each of the two photochemical reaction steps separately, revealing very high quantum yields of 0.16–0.25 for the first step and lower values (0.0055–0.0093) for the second step of the photoreaction. The selective and efficient photochemical reaction makes the photocleavable bis(thioether) ligand scaffold reported here a promising candidate for use in e.g. ruthenium-based photo-activated chemotherapy.

NIR-Light-Driven Generation of Reactive Oxygen Species Using Ru(II)-Decorated Lipid-Encapsulated Upconverting Nanoparticles.

The biological application of ruthenium anticancer prodrugs for photodynamic therapy (PDT) and photoactivated chemotherapy (PACT) is restricted by the need to use poorly penetrating high-energy photons for their activation, i.e., typically blue or green light. Upconverting nanoparticles (UCNPs), which produce high-energy light under near-infrared (NIR) excitation, may solve this issue, provided that the coupling between the UCNP surface and the Ru prodrug is optimized to produce stable nanoconjugates with efficient energy transfer from the UCNP to the ruthenium complex. Herein, we report on the synthesis and photochemistry of the two structurally related ruthenium(II) polypyridyl complexes [Ru(bpy)2(5)](PF6)2 ([1](PF6)2) and [Ru(bpy)2(6)](PF6)2 ([2](PF6)2), where bpy = 2,2-bipyridine, 5 is 5,6-bis(dodecyloxy)-2,9-dimethyl-1,10-phenanthroline, and 6 is 5,6-bis(dodecyloxy)-1,10-phenanthroline. [1](PF6)2 is photolabile as a result of the steric strain induced by ligand 5, but the irradiation of [1](PF6)2 in solution leads to the nonselective and slow photosubstitution of one of its three ligands, making it a poor PACT compound. On the other hand, [2](PF6)2 is an efficient and photostable PDT photosensitizer. The water-dispersible, negatively charged nanoconjugate UCNP@lipid/[2] was prepared by the encapsulation of 44 nm diameter NaYF4:Yb3+,Tm3+ UCNPs in a mixture of 1,2-dioleoyl-sn-glycero-3-phosphate and 1,2-dioleoyl-sn-glycero-3-phosphocholine phospholipids, cholesterol, and the amphiphilic complex [2](PF6)2. A nonradiative energy transfer efficiency of 12% between the Tm3+ ions in the UCNP and the Ru2+ acceptor [2]2+ was found using time-resolved emission spectroscopy. Under irradiation with NIR light (969 nm), UCNP@lipid/[2] was found to produce reactive oxygen species (ROS), as judged by the oxidation of the nonspecific ROS probe 2′,7′-dichlorodihydrofluorescein (DCFH2–). Determination of the type of ROS produced was precluded by the negative surface charge of the nanoconjugate, which resulted in the electrostatic repulsion of the more specific but also negatively charged 1O2 probe tetrasodium 9,10-anthracenediyl-bis(methylene)dimalonate (Na4(ADMBMA)).

Diazido platinum(iv) complexes for photoactivated anticancer chemotherapy

Diazido Pt(iv) complexes with a general formula [Pt(N3)2(L)(L′)(OR)(OR′)] are a new generation of anticancer prodrugs designed for use in photoactivated chemotherapy.

Incorporation of 7-dehydrocholesterol into liposomes as a simple, universal and efficient way to enhance anticancer activity by combining PDT and photoactivated chemotherapy.

Cholesterol (CHOL) is an indispensable component of liposomes. Incorporation of 7-dehydrocholesterol (7-DHC) instead of CHOL can efficiently enhance the anticancer activity of photosensitizer-encapsulated liposomes upon irradiation, yielding an IC50 value about half of that of CHOL-based controls. The photo-oxidation of 7-DHC into its endoperoxide form by singlet oxygen may account for the enhanced therapeutic effect, realizing an efficient combination of photodynamic therapy (PDT) and photoactivated chemotherapy.

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.

Fluorination on non-photolabile dppz ligands for improving Ru(ii) complex-based photoactivated chemotherapy.

Ru(ii) polypyridine complexes which can undergo photo-induced ligand dissociation and subsequent DNA covalent binding may potentially serve as photoactivated chemotherapeutic (PACT) agents. In this paper, three fluorinated dppz ligand coordinated Ru(ii) complexes (2-4) containing four monodentate pyridine ligands were studied. All complexes released one pyridine and covalently bound to DNA upon 470 nm irradiation. Compared with the parent complex [Ru(dppz)(py)4]2+ (1), 2-4 displayed enhanced phototoxicity but diminished dark cytotoxicity, more favorable for PACT application. Complex 3 is the most efficient one with IC50 values of about 8 μM toward HeLa and SKOV-3 cell lines, and also has a much higher IC50 value toward normal L-02 cells. Our results indicate that fluorination on the retaining ligand may be an efficient way to improve the drug activity of Ru(ii) PACT agents.

Photoswitchable Antimetabolite for Targeted Photoactivated Chemotherapy.

The efficacy and tolerability of systemically administered anticancer agents are limited by their off-target effects. Precise spatiotemporal control over their cytotoxic activity would allow improving chemotherapy treatments, and light-regulated drugs are well suited to this purpose. We have developed phototrexate, the first photoswitchable inhibitor of the human dihydrofolate reductase (DHFR), as a photochromic analogue of methotrexate, a widely prescribed chemotherapeutic drug to treat cancer and psoriasis. Quantification of the light-regulated DHFR enzymatic activity, cell proliferation, and in vivo effects in zebrafish show that phototrexate behaves as a potent antifolate in its photoactivated cis configuration and that it is nearly inactive in its dark-relaxed trans form. Thus, phototrexate constitutes a proof-of-concept to design light-regulated cytotoxic small molecules and a step forward to develop targeted anticancer photochemotherapies with localized efficacy and reduced adverse effects.

Dynamics of Oxygen-Independent Photocleavage of Blebbistatin as a One-Photon Blue or Two-Photon Near-Infrared Light-Gated Hydroxyl Radical Photocage

Development of versatile, chemically tunable photocages for photoactivated chemotherapy (PACT) represents an excellent opportunity to address the technical drawbacks of conventional photodynamic therapy (PDT) whose oxygen-dependent nature renders it inadequate in certain therapy contexts such as hypoxic tumors. As an alternative to PDT, oxygen free mechanisms to generate cytotoxic reactive oxygen species (ROS) by visible light cleavable photocages are in demand. Here, we report the detailed mechanisms by which the small molecule blebbistatin acts as a one-photon blue light-gated or two-photon near-infrared light-gated photocage to directly release a hydroxyl radical (•OH) in the absence of oxygen. By using femtosecond transient absorption spectroscopy and chemoselective ROS fluorescent probes, we analyze the dynamics and fate of blebbistatin during photolysis under blue light. Water-dependent photochemistry reveals a critical process of water-assisted protonation and excited state intramolecular proton transfer (ESIPT) that drives the formation of short-lived intermediates, which surprisingly culminates in the release of •OH but not superoxide or singlet oxygen from blebbistatin. CASPT2//CASSCF calculations confirm that hydrogen bonding between water and blebbistatin underpins this process. We further determine that blue light enables blebbistatin to induce mitochondria-dependent apoptosis, an attribute conducive to PACT development. Our work demonstrates blebbistatin as a controllable photocage for •OH generation and provides insight into the potential development of novel PACT agents.

Red‐Light‐Controlled Release of Drug–Ru Complex Conjugates from Metallopolymer Micelles for Phototherapy in Hypoxic Tumor Environments

AbstractTraditional photodynamic phototherapy is not efficient for anticancer treatment because solid tumors have a hypoxic microenvironment. The development of photoactivated chemotherapy based on photoresponsive polymers that can be activated by light in the “therapeutic window” would enable new approaches for basic research and allow for anticancer phototherapy in hypoxic conditions. This work synthesizes a novel Ru‐containing block copolymer for photoactivated chemotherapy in hypoxic tumor environment. The polymer has a hydrophilic poly(ethylene glycol) block and a hydrophobic Ru‐containing block, which contains red‐light‐cleavable (650–680 nm) drug–Ru complex conjugates. The block copolymer self‐assembles into micelles, which can be efficiently taken up by cancer cells. Red light induces release of the drug–Ru complex conjugates from the micelles and this process is oxygen independent. The released conjugates inhibit tumor cell growth even in hypoxic tumor environment. Furthermore, the Ru‐containing polymer for photoactivated chemotherapy in a tumor‐bearing mouse model is applied. Photoactivated chemotherapy of the polymer micelles demonstrates efficient tumor growth inhibition. In addition, the polymer micelles do not cause any toxic side effects to mice during the treatment, demonstrating good biocompatibility of the system to the blood and healthy tissues. The novel red‐light‐responsive Ru‐containing polymer provides a new platform for phototherapy against hypoxic tumors.

Photoactivation of the Cytotoxic Properties of Platinum(II) Complexes through Ligand Photoswitching.

The development of photoactivatable metal complexes with potential anticancer properties is a topical area of current investigation. Photoactivated chemotherapy using coordination compounds is typically based on photochemical processes occurring at the metal center. In the present study, an innovative approach is applied that takes advantage of the remarkable photochemical properties of diarylethenes. Following a proof-of-concept study with two complexes, namely, C1 and C2, a series of additional platinum(II) complexes from dithienylcyclopentene-based ligands was designed and prepared. Like C1 and C2, these new coordination compounds exhibit two thermally stable, interconvertible photoisomers that display distinct properties. The photochemical behavior of ligands L3-L7 has been analyzed by 1H NMR and UV-vis spectroscopies. Subsequently, the corresponding platinum(II) complexes C3-C7 were synthesized and fully characterized, including by single-crystal X-ray diffraction for some of them. Next, the interaction of each photoisomer (i.e., containing the open or closed ligand) of the metal complexes with DNA was examined thoroughly using various techniques, revealing their distinct DNA-binding modes and affinities, as observed for the earlier compounds C1 and C2. The antiproliferative activity of the two forms of the complexes was then assessed with five cancer cell lines and compared with that of C1 and C2, which supported the use of such diarylethene-based systems for the generation of a new class of potential photochemotherapeutic metallodrugs.

Photolabile Ru Model Complexes with Chelating Diimine Ligands for Light‐Triggered Drug Release

A series of water‐soluble photolabile model complexes of the general formula [Ru([9]aneS3)(chel)(py)]Cl2 ([9]aneS3 = 1,4,7‐trithiacyclononane, chel = chelating diimine) was prepared and fully characterized. The phototriggered release of pyridine with visible light as a function of the nature of the diimine {chel = 2,2′‐bipyridine (6) 1,10‐phenanthroline (7), 4,7‐diphenil‐1,10‐phenanthroline (8), dipyrido‐[3,2‐a:2′,3′‐c]phenazine (dppz, 9), 2,2′‐biquinoline (bq, 10)} was investigated. Our aim is to establish whether this type of complexes might be realistically used in the photo‐uncaging strategy of photoactivated chemotherapy (PACT) in the future. Compounds 6–9 present a MLCT absorption in the blue region of the visible spectrum. When irradiated with light at 470 nm, they rapidly and quantitatively release the coordinated pyridine. Complex 10 turned out to be quite different from the others in the series. Structure‐wise, in 10 the average plane of coordinated bq – owing to its steric demand – is remarkably tilted relative to the equatorial coordination plane [37.43(4)°, with the “front“ of the ligand pointing towards the axial py] and the orientation of py is nearly orthogonal compared to that found in 6 and 7 for minimizing steric clashes with bq. The low‐lying acceptor orbitals of bq induce a red‐shift of the MLCT absorption maximum to ca. 500 nm. Contrary to the expectations, complex 10 is more photostable compared to 6–9 and photo‐dissociation of both py and bq, in nearly equal amounts, occurs. A detailed theoretical investigation was performed on 10 (and on 6 for comparison), for explaining its peculiar spectral features and photochemical behavior.

Beyond Photodynamic Therapy: Light-Activated Cancer Chemotherapy.

Cancer chemotherapy is limited by severe side effects due to unspecific cytotoxic activity of currently used therapeutics. In order to minimize these unwanted effects, several approaches have been taken, relying on the use of light to activate drugs. As light can be delivered with a very high spatiotemporal resolution, this technique is a promising strategy to selectively activate cytotoxic drugs at their site of action and thus to improve the tolerability and safety of chemotherapy. This review summarizes different approaches towards photoactivated chemotherapy and identifies its challenges and opportunities. The respective papers were summarized and evaluated in terms of their phototherapeutic indices and the wavelength needed for activation. First, the design, synthesis and/or evaluation of photoactivated metal complexes including platinum- , ruthenium-, and rhodium-complexes is described. Next, photocaged metal complexes and photoacaged organic chemotherapeutics are reported, with a wide range of cytotoxicity mechanisms. The final part includes, examples of photoswitchable drugs for cancer therapy. Some designs, especially metal complexes, stand out due to their very high phototherapeutic index (> 1880) but the common drawback of light-responsive metal complexes and organic chemotherapeutics is the irreversibility of activation. Photoswitchable drugs, however, address this challenge. Nevertheless, the need of UV light for their activation still limits their application. The field of photoactivated cancer chemotherapy is rapidly growing and already includes very promising approaches with designs providing high phototherapeutic indices and also NIR or visible light-activatable drugs.

Photoresponsive ruthenium-containing polymers: potential polymeric metallodrugs for anticancer phototherapy.

This Frontier presents the recent development of photoresponsive Ru-containing polymers for cancer treatment. These novel Ru-containing polymers are prepared by introducing photoresponsive Ru complexes into polymers. Based on their chemical structures in aqueous solutions, these polymers can self-assemble into different nanostructures. The self-assembled nanostructures can circulate in the blood stream, accumulate at tumor tissue, and can be taken up by tumor cells. Red light, which can penetrate into tissue deeply, can induce the photodissociation of these polymers and sensitize singlet oxygen (1O2) generation. Both dissociated Ru complexes and generated 1O2 can inhibit the growth of tumor cells. Photoresponsive Ru-containing polymers provide a new platform for combined photodynamic therapy and photoactivated chemotherapy. The design strategies, self-assembly, photoresponsiveness, and anticancer effects of these polymers are introduced. Some remaining challenges for Ru-containing polymers for phototherapy are discussed.

Why develop photoactivated chemotherapy?

Photoactivated chemotherapy is an approach where a biologically active compound is protected against interaction with the cell environment by a light-cleavable protecting group, and unprotected by light irradiation. As such, PACT represents a major scientific opportunity for developing new bioactive inorganic compounds. However, the societal impact of this approach will only take off if the PACT field is used to address real societal challenges, i.e., therapeutic questions that make sense in a clinical context, rather than purely chemical questions. In particular, I advocate here that the field has become mature enough to switch from a compound-based approach, where a particular cancer model is chosen only to demonstrate the utility of a compound, to a disease-based approach, where the question of which disease to cure comes first: which PACT compound should I make to solve that particular clinical problem? The advantages and disadvantages of PACT vs. other phototherapeutic techniques are discussed, and a roadmap towards real clinical applications of PACT is drawn.

Ruthenium-based PACT compounds based on an N,S non-toxic ligand: a delicate balance between photoactivation and thermal stability

In photoactivated chemotherapy, the photocleavable protecting group that prevents the bioactive compound from interacting with biomolecules in the dark is sometimes cytotoxic, which makes interpretation of phototoxicity challenging. For ruthenium polypyridyl complexes new, non-toxic protecting ligands that prevent a toxic metal complex from binding to biomolecules in the dark, but that can be efficiently photosubstituted upon visible light irradiation to recover the high toxicity of the metal complex, are necessary. In this work, we report on the synthesis, stereochemical characterization and cytotoxicity of a series of polypyridyl complexes; [Ru(bpy)2(mtpa)](PF6)2 ([1](PF6)2, bpy = 2,2’-bipyridine), [Ru(bpy)(dmbpy)(mtpa)](PF6)2 ([2](PF6)2, dmbpy = 6,6’-dimethyl-2,2’-bipyridine), and [Ru(dmbpy)2(mtpa)](PF6)2 ([3](PF6)2) based on the non-toxic 3-(methylthio)propylamine protecting ligand (mtpa). The number of methyl groups had a crucial effect on the photochemistry and cytotoxicity of these complexes. The non-strained complex [1]2+ was not capable of fully releasing mtpa and was not phototoxic in lung cancer cells (A549). In the most strained complex [3]2+, thermal stability was lost, leading to poor photoactivation in vitro and a generally high toxicity also without light activation. The heteroleptic complex [2]2+ with intermediate strain showed, upon blue light irradiation, efficient mtpa photosubstitution and increased cytotoxicity in cancer cells, but photosubstitution was not selective. Overall, fine-tuning of the lipophilicity and steric strain of ruthenium complexes appears as an efficient method to obtain phototoxic ruthenium-based photoactivated chemotherapeutic prodrugs, at the cost of synthetic simplicity and photosubstitution selectivity.