Amphiphilic Photosensitizer Research Articles

Tracking Microenvironmental Response on Self-Assembled Phthalocyanine Systems - Adaptive and Non-Adaptive Antibacterial Photosensitization.

Self-assembly has proven to be one of the effective methods for the formation of nanoscale therapeutics without the need to use nanodelivery systems. Such minimal models of supramolecular systems formed from amphiphilic photosensitizers (PS) have recently emerged as a new class of photoactive systems, providing unique and in some cases superior activities. Although the mechanism of photogenerated reactive oxygen species (ROS) in such systems is studied and to a certain extent understood, there are very limited studies investigating the influence of intricate environmental factors, including those occurring in the cellular environment, on the self-assembly and thus the activity of the system. Understanding the optimal conditions for the formation of active PS aggregates is an important area of research in the field of photodynamic therapy (PDT), as it is directly linked to the optimal treatment dose. In this study, we describe the synthesis, self-assembly properties, photophysical characterization, and photobiological efficacy of structurally closely related low-symmetry phthalocyanine derivatives. Studying the decay behavior of the PS fluorescence lifetime in the presence of molecular crowders and different bacterial strains, we found that certain derivatives exhibited adaptive behavior and change in activity, while others demonstrated non-adaptive characteristics.

Acidity-activatable dynamic hybrid nanoplatforms derived from extracellular vesicles of M1 macrophages enhance cancer immunotherapy through synergistic triple immunotherapy

Immunotherapy exhibits considerable promise for sustained tumor reduction. However, current cancer immunotherapy methods elicit limited responses due to the inadequate immunogenicity exhibited by cancer cells. This obstacle may be addressed using nanoplatforms that can activate synergistic therapies (photodynamic therapy and ferroptosis) in response to the acidic pH of the tumor microenvironment. We previously developed an amphiphilic photosensitizer, SR780, which displays satisfactory photodynamic effects. This photosensitizer is inactivated when bound to Fe3+ (SR780Fe) but is activated upon release in mildly acidic conditions. In this study, M1 macrophage-derived extracellular vesicles (EVs) were fused with REV and SR780Fe–loaded liposomes (REV@SR780Fe@Lip) to form REV@SR780Fe@LEV hybrid nanovesicles. Further modification with the RS17 peptide for tumor targeting enabled a combination of photodynamic therapy, ferroptosis, and cGAS-STING pathway activation, resulting in enhanced antitumor efficacy through a synergistic effect. Upon laser irradiation, REV@SR780Fe@LEV-RS17 demonstrated antitumor effects in 4T1 breast cancer models, including the inhibition of lung and liver metastasis, as well as prevention of tumor recurrence.Graphical

Specific photodamage on HT-29 cancer cells leads to endolysosomal failure and autophagy blockage by cathepsin depletion

Endolysosomes perform a wide range of cellular functions, including nutrient sensing, macromolecule digestion and recycling, as well as plasma membrane repair. Because of their high activity in cancerous cells, endolysosomes are attractive targets for the development of novel cancer treatments. Light-activated compounds termed photosensitizers (PS) can catalyze the oxidation of specific biomolecules and intracellular organelles. To selectively damage endosomes and lysosomes, HT-29 colorectal cancer cells were incubated with nanomolar concentrations of meso-tetraphenylporphine disulfonate (TPPS2a), an amphiphilic PS taken up via endocytosis and activated by green light (522 nm, 2.1 J.cm−1). Several cellular responses were characterized by a combination of immunofluorescence and immunoblotting assays. We showed that TPPS2a photosensitization blocked autophagic flux without extensive endolysosomal membrane rupture. Nevertheless, there was a severe functional failure of endolysosomes due to a decrease in CTSD (cathepsin D, 55%) and CTSB (cathepsin B, 52%) maturation. PSAP (prosaposin) processing (into saposins) was also considerably impaired, a fact that could be detrimental to glycosphingolipid homeostasis. Therefore, photosensitization of HT-29 cells previously incubated with a low concentration of TPPS2a promotes endolysosomal dysfunction, an effect that can be used to improve cancer therapies.

Light-Induced Liposomal Drug Delivery with an Amphiphilic Porphyrin and Its Chlorin and Bacteriochlorin Analogues.

The development of targeted drug delivery mechanisms in the human body is a matter of growing interest in medical science. The selective release of therapeutic agents at a specific target site can increase the therapeutical efficiency and at the same time reduce the side effects. Light-sensitive liposomes can release a drug by an externally controlled light trigger. Liposomes containing photosensitizers that can be activated in the longer wavelength range (650-800 nm) are particularly intriguing for medical purposes. This is because light penetration into a tissue is more efficient within this wavelength range, increasing their potential applications. For this study, liposomes with an encapsulated amphiphilic photosensitizer, the porphyrin 5,10-DiOH (5,10-di(4-hydroxyphenyl)-15,20-diphenyl-21,23H-porphyrin), its chlorin (5,10-DiOH-chlorin) and its bacteriochlorin (5,10-DiOH-bacteriochlorin) were synthesized. The porphyrin 5,10-DiOH showed previously effective cargo release after liposomal encapsulation when irradiated at a wavelength of 420 nm. The new synthesized chlorin and bacteriochlorin photosensitizers show additional absorption bands in the longer wavelength range, which would enable excitation in deeper layers of tissue. Effective cargo release with chlorin at a longer wavelength of 650 nm and bacteriochlorin at 740 nm was possible. Irradiation of chlorin allowed more than 75% of the cargo to be released and more than 60% for bacteriochlorin. The new liposomes would enable selective drug release in deeper tissue layers and expand the range of possible applications.

Amphiphilic photosensitizer polymer as a nanocarrier of cytotoxic molecule for carrier‐free combination therapy

AbstractVarious nanocarriers have been explored to deliver drugs for combination therapy. However, most nanocarriers are composed of inert materials without contribution for improving the cancer therapeutic effect. Herein, a hydrophobic photosensitizer is conjugated to poly (ethylene glycol) to form an amphiphilic polymer, which further self‐assembles into nanomicelle. The generated nanomicelle can act as a nanocarrier to encapsulate a cytotoxic molecule, IR‐775, for combination therapy. The yielded nanodrug is totally composed of pharmacologically active ingredients to avoid any possible toxicity resulted from carrier materials. The nanodrug performs enhanced therapeutic effect compared with any monotherapy and exhibits negligible hemolysis, indicating good biocompatibility for further in vivo applications.

A self-assembled nanophotosensitizer targets lysosomes and induces lysosomal membrane permeabilization to enhance photodynamic therapy.

We report the self-assembly of amphiphilic BDQ photosensitizers into lysosome-targeting nanophotosensitizer BDQ-NP for highly effective photodynamic therapy (PDT). Molecular dynamics simulation, live cell imaging, and subcellular colocalization studies showed that BDQ strongly incorporated into lysosome lipid bilayers to cause continuous lysosomal membrane permeabilization. Upon light irradiation, the BDQ-NP generated a high level of reactive oxygen species to disrupt lysosomal and mitochondrial functions, leading to exceptionally high cytotoxicity. The intravenously injected BDQ-NP accumulated in tumours to achieve excellent PDT efficacy on subcutaneous colorectal and orthotopic breast tumor models without causing systemic toxicity. BDQ-NP-mediated PDT also prevented metastasis of breast tumors to the lungs. This work shows that self-assembled nanoparticles from amphiphilic and organelle-specific photosensitizers provide an excellent strategy to enhance PDT.

Photophysical properties and aggregation behavior of diethylene glycol substituted pyropheophorbide-a in cationic surfactant solutions

The aggregation behavior of a novel amphiphilic photosensitizer - pyropheophorbide-[Formula: see text] 17-diethylene glycol ester 1 was studied in aqueous and aqueous-organic mixtures by means of steady-state absorption and fluorescence emission spectroscopy. The formation of [Formula: see text]-aggregates with a bathochromic shift of the absorption bands, weak fluorescence and resonance enhancement of the light scattering was observed in the mixed water-organic solvents. Solubilization studies in micellar cationic surfactant solutions of alkyl trimethylammonium bromides (C[Formula: see text]TAB) and alkyl triphenylphosphonium bromides (C[Formula: see text]TPPB) resulted in the most effective stabilization of the fluorescent monomolecular form of 1 in C[Formula: see text]TPPB micelles. It was shown that [Formula: see text]-aggregate formation in submicellar surfactant solutions is the initial stage of the solubilization process, which significantly increases in phosphate buffer medium at pH 7.4. Fluorescence lifetime ([Formula: see text] of 1 in DMSO was found to be 7.4 ns, fluorescence quantum yield - 34% and singlet oxygen quantum yield - 62%. The results obtained indicate the need for further studies of compound 1 solubilized in C[Formula: see text]TPPB micelles as the most promising prototype of the drug formulation for mitochondria-targeted photodynamic therapy of cancer.

PH-activated nanoplatform for visualized photodynamic and ferroptosis synergistic therapy of tumors.

To overcome drug resistance and improve precision theranostics for hepatocellular carcinoma (HCC), a nanoplatform with an "off/on" function for multimodality imaging (near-infrared-II (NIR-II) fluorescence imaging, magnetic resonance imaging (MRI), and photoacoustic imaging) and synergistic therapy (photodynamic therapy and ferroptosis) activated by an acidic pH in the tumor microenvironment is proposed. Although many photosensitizers with photodynamic effects have been reported, very few of them have outstanding photodynamic effect and high stability with response to endogenous stimuli capable of NIR-II imaging. Herein, a new amphiphilic photosensitizer SR780 derived from croconaine dye, was developed with satisfactory photodynamic effects and pH-responsive NIR-II imaging. Interestingly, it was deactivated by coordination with Fe3+ (SR780@Fe) and activated during their release under mild acidic condition. Ferroptosis can generate hydroxyl free radical and lipid peroxide, which aggravate the oxidative stress of tumor cells and mediate their death while depleting glutathione (GSH) to enhance photodynamic effect. In situ pH-activatable theranostic nanoplatform, SR780@Fe-PAE-GP, was thus developed by loading SR780@Fe with pH-responsive polymers, modified by a glypican-3 (GPC-3) receptor-targeting peptide. The synergistic antitumor effects were confirmed both in vitro and in vivo, and the tumor inhibition rate of the SR780@Fe-PAE-GP+L treatment group reached 98%.

Interfacial Characterization of Ruthenium-Based Amphiphilic Photosensitizers.

Nonreactive surfactant molecules have long been used and characterized for a wide range of applications in industries, life science, and everyday life. Recently, new types of functional amphiphilic molecules have emerged that bear another function, for example, a light-absorbing action, or catalytic properties. However, the surfactant properties of these molecules remain to date essentially unknown. In this context, we investigated here the interfacial activity of photocatalytic surfactants based on a ruthenium(II) tris-bipyridine core, functionalized with two alkyl tails. We realized a systematic characterization of the surfactant properties of these molecules at a water–air interface and studied the effect of the alkyl chain length and of the counterions (hexafluorophosphate or chloride) on these properties. Our data demonstrate that ruthenium surfactants with chloride counteranions form a denser layer at the interface, but their surfactant properties can dramatically deteriorate when the chain length of the alkyl tail increases, leading to simple hydrophobic molecules with poor surfactant properties for the longest chains (C17). These findings pave the way for a better use and understanding of photocatalytic soft interfaces.

Self-Assembled Liposomes Enhance Electron Transfer for Efficient Photocatalytic CO2 Reduction.

Light-driven conversion of CO2 to chemicals provides a sustainable alternative to fossil fuels, but homogeneous systems are typically limited by cross reactivity between different redox half reactions and inefficient charge separation. Herein, we present the bioinspired development of amphiphilic photosensitizer and catalyst pairs that self-assemble in lipid membranes to overcome some of these limitations and enable photocatalytic CO2 reduction in liposomes using precious metal-free catalysts. Using sodium ascorbate as a sacrificial electron source, a membrane-anchored alkylated cobalt porphyrin demonstrates higher catalytic CO production (1456 vs 312 turnovers) and selectivity (77 vs 11%) compared to its water-soluble nonalkylated counterpart. Time-resolved and steady-state spectroscopy revealed that self-assembly facilitates this performance enhancement by enabling a charge-separation state lifetime increase of up to two orders of magnitude in the dye while allowing for a ninefold faster electron transfer to the catalyst. Spectroelectrochemistry and density functional theory calculations of the alkylated Co porphyrin catalyst support a four-electron-charging mechanism that activates the catalyst prior to catalysis, together with key catalytic intermediates. Our molecular liposome system therefore benefits from membrane immobilization and provides a versatile and efficient platform for photocatalysis.

Influence of Parameters on Photodynamic Therapy of Au@TiO2-HMME Core-Shell Nanostructures.

Photodynamic therapy (PDT) is a promising tumor therapy and has been proven to be an effective, safe and minimally invasive technique. Hematoporphyrin monomethyl ether (HMME) mediated PDT has been used in clinical treatment of port wine stain (PWS) due to its single component, high yield of singlet oxygen and short light-sensitive period. However, as an amphiphilic photosensitizer, HMME is easy to aggregate due to the presence of a hydrophobic group, which undesirably reduced its generation of singlet oxygen and bioavailability. In this study, we synthesized the stable conjugate of Au@TiO2 core-shell nanostructure with HMME, and the influence of different factors on PTD efficiency were studied. The results showed that the nanostructure had higher PTD efficiency for KB cells than that of HMME. The irradiation wavelength, gold nanoparticle shape and the shell thickness are all important factors for KB cell PDT.

Engineering Cellular Membrane for Dual Mode Therapy Using NIR Responsive Photosensitizer and Reversible Topoisomerase Inhibition Activity.

Extensive research over past few decades has highlighted the challenges of chemotherapy and prompted the need for multimodality therapy because chemotherapy alone cannot fully eradicate the tumor due to physiological barriers in its effective delivery and systemic side effects. It can be mitigated by adopting nanoparticles as more effective delivery method, but none of them completely prevents drug toxicities. Utilizing multiple therapeutic modes such as phototherapy that can act synergistically with chemotherapy in controlling tumor growth, while reducing the overall dosage, could become a preferred route for cancer management. Careful selection of nanoparticle system, which can simultaneously deliver both drug and photosensitizer, can significantly enhance the therapeutic outcome. Therefore, in this paper, we report development and potential of immune-compatible and long circulating nanoerythrosomes for enhancing the therapeutic potential of camptothecin and indocyanine green against murine cancer model. The RBCs membrane simultaneously loaded the nonpolar drug and amphiphilic photosensitizer in its lipid bilayer, which self-assembled to form stable nanoparticles. These nano constructs absorbed light in the near-infrared region and hence are suitable for targeting deep seated tissues. The dual chemo-phototherapy had great effect on cell viability and had therapeutic value.

Biosupramolecular complexes of amphiphilic photosensitizers with human serum albumin and cucurbit[7]uril as carriers for photodynamic therapy.

In the present work, we evaluated the supramolecular interactions between three photosensitizers, namely toluidine blue O (TBO, positively charged) and two fatty acid conjugates of 6 and 14 carbon atoms chain lengths (TBOC6 and TBOC14), with human serum albumin (HSA) and the macrocycle cucurbit[7]uril (CB[7]), alone or in combination within a biosupramolecular system as potential carriers of photosensitizers for Photodynamic therapy (PDT). Binding studies were carried out using photophysical and calorimetric techniques and accompanied with molecular docking simulations. Amphiphilic photosensitizers, particularly TBOC14, showed stronger binding to HSA and (CB[7]). Comparing the different delivery systems, (CB[7]) had a marginal effect on cell uptake and phototoxicity in HeLa cells, while HSA showed enhanced cell uptake with phototoxicities that depended on the photosensitizer. Despite low cell uptake, the combination of both (CB[7]) and HSA was the most phototoxic, which illustrates the potential of combining these systems for PDT applications.

Multifunctional photo-responsive liposomes for tumor imaging and phototherapy to enhance the antitumor efficacy and reduce the hepatotoxicity of methotrexate

Methotrexate (MTX) is a chemotherapy drug widely applied in clinical treatment for various tumors. However, side effects caused by MTX greatly affect the therapeutic efficacy. Hence, photo-triggered release liposomes were constructed by amphiphilic photosensitizers ZnPc(PEG)4 and phospholipid materials DPPC to load MTX for improved antitumor effect. It was demonstrated that ZnPc(PEG)4 integrated into lipid layers endowed final product ZnPc(PEG)4:MTX@LiPOs improved stability. ZnPc(PEG)4:MTX@LiPOs was verified to have excellent characteristics in terms of photo-triggered release, ROS generation and heat production. In particular, cellular experiments indicated that ZnPc(PEG)4:MTX@LiPOs had synergistic cytotoxicity of phototherapy and chemotherapy. The cell uptake assay demonstrated that the internalization of MTX into tumor cells was significantly increased by the encapsulation of ZnPc(PEG)4@LiPOs. ZnPc(PEG)4:MTX@LiPOs was confirmed to possess excellent tumor targeting reflected in faster accumulation, higher drug level and longer retention at tumor sites than free ZnPc(PEG)4. In addition, ZnPc(PEG)4:MTX@LiPOs had significantly enhanced tumor inhibition rate. What's more, liver histopathology and serum enzyme tests indicated that ZnPc(PEG)4:MTX@LiPOs had distinctly reduced hepatotoxicity than MTX. The constructed liposomes with photo-triggered release can be applied to deliver other chemotherapy drugs for synergistic antitumor ability and reduced side effect.

Self-assembly of Amphiphilic Porphyrins To Construct Nanoparticles for Highly Efficient Photodynamic Therapy.

Hydrophobic photosensitizers greatly affect cell permeability and enrichment in tumors, but they cannot be used directly for clinical applications because they always aggregate in water, preventing their circulation in the blood and accumulation in tumor cells. As a result, amphiphilic photosensitizers are highly desirable. Although nanomaterial-based photosensitizers can solve water solubility, they have the disadvantages of complicated operation, poor reproducibility, low drug loading, and poor stability. In this work, an efficient synthesis strategy is proposed that converts small molecules into nanoparticles in 100 % aqueous solution by molecular assembly without the addition of any foreign species. Three photosensitizers with triphenylphosphine units and ethylene glycol chains of different lengths, TPP-PPh3 , TPP-PPh3 -2PEG and TPP-PPh3 -4PEG, were synthesized to improve amphiphilicity. Of the three photosensitizers, TPP-PPh3 -4PEG is the most efficient (singlet oxygen yield: 0.89) for tumor photodynamic therapy not only because of its definite constituent, but also because its amphiphilic structure allows it to self-assemble in water.

Self-Assembled Porphyrinoids: One-Component Nanostructured Photomedicines.

Photodynamic therapy (PDT) is becoming a promising way to treat various kinds of cancers, with few side effects. Porphyrinoids are the most relevant photosensitizers (PS) in PDT, because they present high extinction coefficients, biocompatibility, and excellent photochemical behavior. To maximize therapeutic effects, polymer‐PS conjugates, and PS‐loaded nanoparticles have been developed, with insights in improving tumor delivery. However, some drawbacks such as non‐biodegradability, multistep fabrication, and low reagent loadings limit their clinical application. A novel strategy, noted by some authors as the “one‐for‐all” approach, is emerging to circumvent the use of additional delivery agents. This approach relies on the self‐assembly of amphiphilic PS to fabricate nanostructures with improved transport properties. In this review we focus on different rational designs of porphyrinoid PS to achieve some of the following attributes in nanoassembly: i) selective uptake, through the incorporation of recognizable biological vectors; ii) responsiveness to stimuli; iii) combination of imaging and therapeutic functions; and iv) multimodal therapy, including photothermal or chemotherapy abilities.

Cytoplasmic delivery of small interfering RNA by photoresponsive non-cationic liposomes

Small interfering RNA (siRNA) can specifically suppress gene expression by cleaving mRNA in the cytoplasm, termed RNA interference (RNAi). Although a nanoparticle-based siRNA delivery system has been approved for clinical use, the efficiency of cytoplasmic delivery of siRNA is still low. Recently, our group has established a highly efficient siRNA encapsulation method using non-cationic liposomes (LPs), which are more biocompatible than conventional cationic LPs. While non-cationic LPs containing siRNA were taken up by cells in vitro, the cytoplasmic release of siRNA and subsequent phenomena involving RNAi were not observed. It was considered that cytoplasmic delivery of siRNA, and siRNA release from LPs, should occur sequentially. We utilized amphiphilic photosensitizer Al (III) phthalocyanine chloride disulfonic acid (AlPcS2a), which can generate singlet oxygen under exposure to red light. When SK-HEP-1 cells expressing luciferase were treated with non-cationic LPs containing both siRNA and AlPcS2a, RNAi could be observed upon red light exposure, suggesting that singlet oxygen efficiently disrupts the membranous structures of endo/lysosomes and LPs. Thus, the combined use of AlPcS2a and red light is effective for utilizing non-cationic LPs for cytoplasmic delivery of siRNA.

Fatty Acid Conjugates of Toluidine Blue O as Amphiphilic Photosensitizers: Synthesis, Solubility, Photophysics and Photochemical Properties†.

Toluidine blue O (TBO) is a water-soluble photosensitizer that has been used in photodynamic antimicrobial and anticancer treatments, but suffers from limited solubility in hydrophobic media. In an effort to incrementally increase TBO's hydrophobicity, we describe the synthesis of hexanoic (TBOC6) and myristic (TBOC14) fatty acid derivatives of TBO formed in low to moderate percent yields by condensation with the free amine site. Covalently linking 6 and 14 carbon chains led to modifications of not only TBO's solubility, but also its photophysical and photochemical properties. TBOC6 and TBOC14 derivatives were more soluble in organic solvents and showed hypsochromic shifts in their absorption and emission bands. The solubility in phosphate buffer solution was low for both TBOC6 and TBOC14, but unexpectedly slightly greater in the latter. Both TBOC6 and TBOC14 showed decreased triplet excited-state lifetimes and singlet oxygen quantum yields in acetonitrile, which was attributed to heightened aggregation of these conjugates particularly at high concentrations due to the hydrophobic "tails." While in diluted aqueous buffer solution, indirect measurements showed similar efficiency in singlet oxygen generation for TBOC14 compared to TBO. This work demonstrates a facile synthesis of fatty acid TBO derivatives leading to amphiphilic compounds with a delocalized cationic "head" group and hydrophobic "tails" for potential to accumulate into biological membranes or membrane/aqueous interfaces in PDT applications.

Enhanced Cellular Uptake and Photodynamic Effect with Amphiphilic Fluorinated Porphyrins: The Role of Sulfoester Groups and the Nature of Reactive Oxygen Species

A class of amphiphilic photosensitizers for photodynamic therapy (PDT) was developed. Sulfonate esters of modified porphyrins bearing—F substituents in the ortho positions of the phenyl rings have adequate properties for PDT, including absorption in the red, increased cellular uptake, favorable intracellular localization, low cytotoxicity, and high phototoxicity against A549 (human lung adenocarcinoma) and CT26 (murine colon carcinoma) cells. Moreover, the role of type I and type II photochemical processes was assessed by fluorescent probes specific for various reactive oxygen species (ROS). The photodynamic effect is improved not only by enhanced cellular uptake but also by the high generation of both singlet oxygen and oxygen-centered radicals. All of the presented results support the idea that the rational design of photosensitizers for PDT can be further improved by better understanding the determinants affecting its therapeutic efficiency and explain how smart structural modifications can make them suitable photosensitizers for application in PDT.

In situ self-assembled biosupramolecular porphyrin nanofibers for enhancing photodynamic therapy in tumors.

Due to the complicated environment and high tissue hydraulic pressure in tumors that easily pumps the nanomedicines back to the systemic circulation, the concentration of released photosensitizers (PSs) retained in a tumor by a traditional nano-delivery system is very low, causing an unsatisfactory photodynamic therapy (PDT) effect. Therefore, we prepared a pH/H2O2-responsive nano-system (ZnP-OC-M) through modified porphyrin PS units with a long-unsaturated oleoyl chloride chain, and by the further introduction of hydrophilic hydroxyl groups and MnO2 through a cis-addition reaction between the unsaturated double bonds of oleoyl chloride and dilute KMnO4 solution. Making use of the sensitivity of MnO2 to the H2O2 in the acid environment of tumor cells, ZnP-OC-M selectively realized responsive disintegration and O2 generation. More importantly, the rich amphiphilic PS units were shedded simultaneously and spontaneously completed the self-assembly into nanofibers in situ by helical stacking, which displayed a 1.85-fold higher retention effect of PSs in vivo compared with free PS groups and showed a great tumor inhibition effect in enhancing PDT. This nanosystem effectively solves the problem of the low retention abilities leading to a poor PS concentration in a tumor, prolonging the treatment time window efficiently after only a single administration and achieving the purpose of PDT enhancement.