RESUMO
Photodynamic therapy (PDT) utilizes the potential of photosensitizing substances to absorb light energy and produce reactive oxygen species. Tetra-cationic porphyrins, which have organic or coordination compounds attached to their periphery, are heterocyclic derivatives with well-described antimicrobial and antitumoral properties. This is due to their ability to produce reactive oxygen species and their photobiological properties in solution. Consequently, these molecules are promising candidates as new and more effective photosensitizers with biomedical, environmental, and other biomedical applications. Prior to human exposure, it is essential to establish the toxicological profile of these molecules using in vivo models. In this study, we used Caenorhabditis elegans, a small free-living nematode, as a model for assessing toxic effects and predicting toxicity in preclinical research. We evaluated the toxic effects of porphyrins (neutral and tetra-cationic) on nematodes under dark/light conditions. Our findings demonstrate that tetra-methylated porphyrins (3TMeP and 4TMeP) at a concentration of 3.3⯵g/mL (1.36 and 0.93⯵M) exhibit high toxicity (as evidenced by reduced survival, development, and locomotion) under dark conditions. Moreover, photoactivated tetra-methylated porphyrins induce higher ROS levels compared to neutral (3TPyP and 4TPyP), tetra-palladated (3PdTPyP and 4PdTPyP), and tetra-platinated (3PtTPyP and 4PtTPyP) porphyrins, which may be responsible for the observed toxic effects.
Assuntos
Caenorhabditis elegans , Luz , Fármacos Fotossensibilizantes , Porfirinas , Animais , Caenorhabditis elegans/efeitos dos fármacos , Porfirinas/toxicidade , Porfirinas/química , Fármacos Fotossensibilizantes/toxicidade , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/química , Espécies Reativas de Oxigênio/metabolismo , Fotoquimioterapia/métodos , Cátions/toxicidade , Relação Dose-Resposta a DrogaRESUMO
Chronic wound healing, impeded by bacterial infections and drug resistance, poses a threat to global human health. Antibacterial phototherapy is an effective way to fight microbial infection without causing drug resistance. Covalent organic frameworks (COFs) are a class of highly crystalline functional porous carbon-based materials composed of light atoms (e.g., carbon, nitrogen, oxygen, and borane), showing potential applications in the biomedical field. Herein, we constructed porphyrin-based COF nanosheets (TP-Por CON) for synergizing photodynamic and photothermal therapy under red light irradiation (e.g., 635 nm). Moreover, a nitric oxide (NO) donor molecule, BNN6, was encapsulated into the pore volume of the crystalline porous framework structure to moderately release NO triggered by red light irradiation for realizing gaseous therapy. Therefore, we successfully synthesized a novel TP-Por CON@BNN6-integrated heterojunction for thoroughly killing Gram-negative bacteria Escherichia coli and Gram-positive bacteria Staphylococcus aureus in vitro. Our research identified that TP-Por CON@BNN6 has favorable biocompatibility and biodegradability, low phototoxicity, anti-inflammatory properties, and excellent mice wound healing ability in vivo. This study indicates that the TP-Por CON@BNN6-integrated heterojunction with multifunctional properties provides a potential strategy for COF-based gaseous therapy and microorganism-infected chronic wound healing.
Assuntos
Anti-Inflamatórios/uso terapêutico , Estruturas Metalorgânicas/uso terapêutico , Doadores de Óxido Nítrico/uso terapêutico , Fármacos Fotossensibilizantes/uso terapêutico , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Cicatrização/efeitos dos fármacos , Animais , Anti-Inflamatórios/efeitos da radiação , Anti-Inflamatórios/toxicidade , Linhagem Celular , Escherichia coli/efeitos dos fármacos , Luz , Estruturas Metalorgânicas/efeitos da radiação , Estruturas Metalorgânicas/toxicidade , Camundongos Endogâmicos BALB C , Doadores de Óxido Nítrico/efeitos da radiação , Doadores de Óxido Nítrico/toxicidade , Fármacos Fotossensibilizantes/efeitos da radiação , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/efeitos da radiação , Porfirinas/uso terapêutico , Porfirinas/toxicidade , Staphylococcus aureus/efeitos dos fármacosRESUMO
Overproduction of reactive oxygen species (ROS) within tumors can cause oxidative stress on tumor cells to induce death, which has motivated us to develop ROS-mediated tumor therapies, such as typical photodynamic therapy (PDT) and Fenton reaction-mediated chemodynamic therapy (CDT). However, these therapeutic modalities suffer from compromised treatment efficacy owing to their limited generation of highly reactive ROS in a tumor microenvironment (TME). In this work, a nanoscale iron-based metal-organic framework, MIL-101(Fe), is synthesized as a Fenton nanocatalyst to perform the catalytic conversion of hydroxyl radicals (·OH) from hydrogen peroxide (H2O2) under the acidic environment and as a biocompatible and biodegradable nanocarrier to deliver a 5,10,15,20-tetrakis(4-carboxyphenyl)porphyrin (TCPP) photosensitizer for light-activated singlet oxygen (1O2) generation. By coupling such chemodynamic/photodynamic effects, the photosensitizer-integrated nanoagents (MIL-101(Fe)@TCPP) could enable more ROS production within tumors to induce amplified oxidative damage for tumor-specific synergistic therapy. In vitro results show that MIL-101(Fe)@TCPP nanoagents achieve the acid-responsive CDT and effective PDT, and synergistic CDT/PDT provides an enhanced therapeutic effect. Ultimately, based on such synergistic therapy, MIL-101(Fe)@TCPP nanoagents cause a significant tumor growth inhibition in vivo without severe side effects, showing great potential for anti-tumor application.
Assuntos
Antineoplásicos/uso terapêutico , Estruturas Metalorgânicas/uso terapêutico , Nanoestruturas/uso terapêutico , Neoplasias/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Fármacos Fotossensibilizantes/uso terapêutico , Antineoplásicos/química , Antineoplásicos/farmacologia , Antineoplásicos/toxicidade , Catálise , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Peróxido de Hidrogênio/química , Radical Hidroxila/metabolismo , Ferro/química , Luz , Estruturas Metalorgânicas/química , Estruturas Metalorgânicas/farmacologia , Estruturas Metalorgânicas/toxicidade , Nanoestruturas/química , Nanoestruturas/toxicidade , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/efeitos da radiação , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/farmacologia , Porfirinas/efeitos da radiação , Porfirinas/uso terapêutico , Porfirinas/toxicidade , Oxigênio Singlete/metabolismoRESUMO
Due to conventional photodynamic therapy encountering serious problems of phototoxicity and low tissue-penetrating depth of light, other dynamic therapy-based therapeutic methods such as sonodynamic therapy (SDT) are expected to be developed. To improve the therapeutic response to SDT, more effective sonosensitizers are imperative. In this study, a novel water-soluble iridium(III)-porphyrin sonosensitizer (IrTMPPS) was synthesized and used for SDT. IrTMPPS generated ample singlet oxygen (1O2) under US irradiation and especially showed distinguished US-activatable abilities at more than 10 cm deep-tissue depths. Interestingly, under US irradiation, IrTMPPS sonocatalytically oxidized intracellular NADH, which would enhance SDT efficiency by breaking the redox balance in the tumor. Moreover, IrTMPPS displayed great sonocytotoxicity toward various cancer cells, and in vivo experiments demonstrated efficient tumor inhibition and anti-metastasis to the lungs in the presence of IrTMPPS and US irradiation. This report gives a novel idea of metal-based sonosensitizers for sonotherapy by fully taking advantage of non-invasiveness, water solubility, and deep tumor therapy.
Assuntos
Antineoplásicos/uso terapêutico , Neoplasias/tratamento farmacológico , Porfirinas/uso terapêutico , Radiossensibilizantes/uso terapêutico , Animais , Antineoplásicos/síntese química , Antineoplásicos/toxicidade , Linhagem Celular Tumoral , Irídio/química , Irídio/uso terapêutico , Irídio/toxicidade , Camundongos , NAD/química , NAD/metabolismo , Neoplasias/patologia , Oxirredução , Porfirinas/síntese química , Porfirinas/toxicidade , Radiossensibilizantes/síntese química , Radiossensibilizantes/toxicidade , Oxigênio Singlete/metabolismo , Ondas Ultrassônicas , Peixe-ZebraRESUMO
In this study, a family of porphyrins based on 5,10,15,20-Tetrakis(4-ethylphenyl)porphyrin (1, Ph) and six metallo-derivatives (Zn2+(2, Ph-Zn), Sn4+(3, Ph-Sn), Mn2+ (4, Ph-Mn), Ni2+ (5, Ph-Ni), Al3+ (6, Ph-Al), and V3+ (7, Ph-V)) were tested as photosensitizers for photodynamic therapy against Leishmania braziliensis and panamensis. The singlet oxygen quantum yield value (ΦΔ) for (1-7) was measured using 1,3-diphenylisobenzofuran (DPBF) as a singlet oxygen trapping agent and 5,10,15,20-(tetraphenyl)-porphyrin (H2TPP) as a reference standard; besides, parasite viability was estimated by the MTT assay. After metal insertion into the porphyrin core, the ΦΔ increased from 0.76-0.90 and cell viability changed considerably. The ΦΔ and metal type changed the cytotoxic activity. Finally, (2) showed both the highest ΦΔ (0.90) and the best photodynamic activity against the parasites studied (IC50 of 1.2 µM).
Assuntos
Leishmania braziliensis/efeitos dos fármacos , Leishmania/efeitos dos fármacos , Metaloporfirinas/química , Metaloporfirinas/toxicidade , Fármacos Fotossensibilizantes/química , Porfirinas/química , Oxigênio Singlete/química , Benzofuranos/química , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Concentração Inibidora 50 , Leishmania/efeitos da radiação , Leishmania braziliensis/efeitos da radiação , Luz , Metaloporfirinas/síntese química , Fotoquimioterapia , Porfirinas/síntese química , Porfirinas/toxicidadeRESUMO
The discovery of aggregation inhibitors and the elucidation of their mechanism of action are key in the quest to mitigate the toxic consequences of amyloid formation. We have previously characterized the antiamyloidogenic mechanism of action of sodium phtalocyanine tetrasulfonate ([Na4(H2PcTS)]) on α-Synuclein (αS), demonstrating that specific aromatic interactions are fundamental for the inhibition of amyloid assembly. Here we studied the influence that metal preferential affinity and peripheral substituents may have on the activity of tetrapyrrolic compounds on αS aggregation. For the first time, our laboratory has extended the studies in the field of the bioinorganic chemistry and biophysics to cellular biology, using a well-established cell-based model to study αS aggregation. The interaction scenario described in our work revealed that both N- and C-terminal regions of αS represent binding interfaces for the studied compounds, a behavior that is mainly driven by the presence of negatively or positively charged substituents located at the periphery of the macrocycle. Binding modes of the tetrapyrrole ligands to αS are determined by the planarity and hydrophobicity of the aromatic ring system in the tetrapyrrolic molecule and/or the preferential affinity of the metal ion conjugated at the center of the macrocyclic ring. The different capability of phthalocyanines and meso-tetra (N-methyl-4-pyridyl) porphine tetrachloride ([H2PrTPCl4]) to modulate αS aggregation in vitro was reproduced in cell-based models of αS aggregation, demonstrating unequivocally that the modulation exerted by these compounds on amyloid assembly is a direct consequence of their interaction with the target protein.
Assuntos
Proteínas Amiloidogênicas/metabolismo , Indóis/metabolismo , Porfirinas/metabolismo , Multimerização Proteica/efeitos dos fármacos , Zinco/metabolismo , alfa-Sinucleína/metabolismo , Sequência de Aminoácidos , Proteínas Amiloidogênicas/química , Linhagem Celular Tumoral , Complexos de Coordenação/química , Complexos de Coordenação/metabolismo , Humanos , Interações Hidrofóbicas e Hidrofílicas , Indóis/química , Indóis/toxicidade , Porfirinas/química , Porfirinas/toxicidade , Ligação Proteica , Zinco/química , alfa-Sinucleína/químicaRESUMO
Hyaluronic acid (HA)-based nanocarriers are of great interest in the drug delivery field due to the tumor targetability via CD44-mediated recognition and endocytosis. However, sufficient tumor-specific release of encapsulated cargoes with steady controllability is necessary to optimize their outcome for cancer therapy. In this study, we constructed a light-activated nanocarrier TKHCENPDOX to enable on-demand drug release at the desired site (tumor). Particularly, TKHCENPDOX encapsulating doxorubicin (DOX) was self-assembled from a HA-photosensitizer conjugate (HA-TK-Ce6) containing reactive oxygen species (ROS)-sensitive thioketal (TK) linkers. Following i.v. injection, TKHCENPDOX was accumulated in the MDA-MB-231 breast tumor xenograft more efficiently through preventing drug leakage in the bloodstream and the HA-mediated targeting effect. Upon internalization into tumoral cells, 660 nm laser irradiation generated ROS during a photodynamic (PDT) process to cleave the TK linker next to Ce6, resulting in light-induced TKHCENPDOX dissociation and selective DOX release in the tumor area. Consequently, TKHCENPDOX showed a remarkable therapeutic effect and minimized toxicity in vivo. This strategy might provide new insight for designing cancer-selective nanoplatforms with active targeting and locoregional drug release simultaneously.
Assuntos
Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/uso terapêutico , Portadores de Fármacos/química , Ácido Hialurônico/química , Nanoconjugados/química , Animais , Antineoplásicos/farmacocinética , Linhagem Celular Tumoral , Clorofilídeos , Doxorrubicina/farmacocinética , Portadores de Fármacos/efeitos da radiação , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos/efeitos da radiação , Feminino , Humanos , Ácido Hialurônico/efeitos da radiação , Ácido Hialurônico/toxicidade , Luz , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos ICR , Nanoconjugados/efeitos da radiação , Nanoconjugados/toxicidade , Nanopartículas/química , Nanopartículas/efeitos da radiação , Nanopartículas/toxicidade , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/efeitos da radiação , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/farmacologia , Porfirinas/efeitos da radiação , Porfirinas/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Antibacterial photocatalytic therapy has been reported as a promising alternative water disinfection technology for combating antibiotic-resistant bacteria. Numerous inorganic nanosystems have been developed as antibiotic replacements for bacterial infection treatment, but these are limited due to the toxicity risk of heavy metal species. Organic semiconductor photocatalytic materials have attracted great attention due to their good biocompatibility, chemically tunable electronic structure, diverse structural flexibility, suitable band gap, low cost, and the abundance of the resources they require. An all-organic composite photocatalytic nanomaterial C3 N4 /perylene-3,4,9,10-tetracarboxylic diimide (PDINH) heterostructure is created through recrystallization of PDINH on the surface of C3 N4 in situ, resulting in enhanced photocatalytic efficiency due to the formation of a basal heterostructure. The absorption spectrum of this composite structure can be extended from ultraviolet to near-infrared light (750 nm), enhancing the photocatalytic effect to produce more reactive oxygen species, which have an excellent inactivation effect on both Gram-negative and positive bacteria, while demonstrating negligible toxicity to normal tissue cells. An efficient promotion of infectious wound regeneration in mice with Staphylococcus aureus infected dermal wounds is demonstrated. This all-organic heterostructure shows great promise for use in wound disinfection.
Assuntos
Antibacterianos/química , Imidas/química , Nitrilas/química , Perileno/análogos & derivados , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/química , Semicondutores , Animais , Antibacterianos/uso terapêutico , Antibacterianos/toxicidade , Catálise , Sobrevivência Celular , Escherichia coli/efeitos dos fármacos , Feminino , Luz , Camundongos , Camundongos Endogâmicos BALB C , Células NIH 3T3 , Nanoestruturas/química , Nitrilas/uso terapêutico , Nitrilas/toxicidade , Perileno/química , Fármacos Fotossensibilizantes/uso terapêutico , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/uso terapêutico , Porfirinas/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Cicatrização , Infecção dos Ferimentos/tratamento farmacológico , Infecção dos Ferimentos/microbiologiaRESUMO
We demonstrate a MnO2-based nanoreactor to achieve continuous oxygen generation and efficient conversion from glucose to singlet oxygen for combined photodynamic-starvation therapy.
Assuntos
Membrana Celular/química , Compostos de Manganês/uso terapêutico , Nanopartículas/uso terapêutico , Neoplasias/tratamento farmacológico , Óxidos/uso terapêutico , Fotoquimioterapia/métodos , Animais , Linhagem Celular Tumoral , Clorofilídeos , Enzimas Imobilizadas/química , Enzimas Imobilizadas/uso terapêutico , Enzimas Imobilizadas/toxicidade , Feminino , Glucose/química , Glucose Oxidase/química , Glucose Oxidase/uso terapêutico , Glucose Oxidase/toxicidade , Peróxido de Hidrogênio/química , Compostos de Manganês/química , Camundongos Endogâmicos BALB C , Nanopartículas/química , Nanopartículas/toxicidade , Óxidos/química , Óxidos/toxicidade , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/uso terapêutico , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/química , Porfirinas/uso terapêutico , Porfirinas/toxicidade , Oxigênio Singlete/química , Hipóxia Tumoral/efeitos dos fármacosRESUMO
Photodynamic therapy (PDT) involves a photosensitizing agent activated with light to induce cell death. Two-photon excited PDT (TPE-PDT) offers numerous benefits compared to traditional one-photon induced PDT, including an increased penetration depth and precision. However, the in vitro profiling and comparison of two-photon photosensitizers (PS) are still troublesome. Herein, we report the development of an in vitro screening platform of TPE-PS using a 3D osteosarcoma cell culture. The platform was tested using three different two-photon (2P) active compounds - a 2P sensitizer P2CK, a fluorescent dye Eosin Y, and a porphyrin derivative (TPP). Their 2P absorption cross-sections (σ2PA) were characterised using a fully automated z-scan setup. TPP exhibited a remarkably high σ2PA at 720 nm (8865 GM) and P2CK presented a high absorption at 850 nm (405 GM), while Eosin Y had the lowest 2P absorption at the studied wavelengths (<100 GM). The cellular uptake of PS visualized using confocal laser scanning microscopy showed that both TPP and P2CK were internalized by the cells, while Eosin Y stayed mainly in the surrounding media. The efficiency of the former two TPE-PS was quantified using the PrestoBlue metabolic assay, showing a significant reduction in cell viability after two-photon irradiation. The possibility of damage localization was demonstrated using a co-culture of adipose derived stem cells together with osteosarcoma spheroids showing no signs of damage to the surrounding healthy cells after TPE-PDT.
Assuntos
Antineoplásicos/farmacologia , Compostos de Benzilideno/farmacologia , Amarelo de Eosina-(YS)/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , Antineoplásicos/efeitos da radiação , Antineoplásicos/toxicidade , Compostos de Benzilideno/efeitos da radiação , Compostos de Benzilideno/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/farmacologia , Amarelo de Eosina-(YS)/efeitos da radiação , Amarelo de Eosina-(YS)/toxicidade , Humanos , Células-Tronco Mesenquimais , Osteossarcoma/tratamento farmacológico , Fótons , Fármacos Fotossensibilizantes/efeitos da radiação , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/efeitos da radiação , Porfirinas/toxicidadeRESUMO
Nanocarriers for drug delivery have made great progress in the treatment of cancer, but the dense extracellular collagen of tumors has greatly limited the efficiency of drug delivery. In this study, losartan is used to deplete tumor collagen and improve the delivery efficiency and photodynamic therapeutic efficacy of chlorine 6 (Ce6)-loaded periodic mesoporous organosilica nanoplatform (Ce6-PMO) for breast cancer. After pretreatment with losartan in vivo, the tumor collagen I fraction is significantly reduced by 53% compared to that of mice pretreated with saline. Importantly, the accumulation of the Ce6-PMO nanoplatforms in the tumor is remarkably enhanced via peritumoral and intravenous injection, respectively, after the mice are pretreated with losartan. Further, combination of losartan with the Ce6-PMO nanoplatforms shows the best therapeutic efficacy, and the suppression rate of tumor volume is measured up to 82%. Taken together, this study provides a very promising synergetic strategy to improve the tumor photodynamic treatment efficacy of nanoplatforms.
Assuntos
Colágeno Tipo I/metabolismo , Losartan/administração & dosagem , Neoplasias Mamárias Experimentais/tratamento farmacológico , Fotoquimioterapia , Porfirinas/administração & dosagem , Radiossensibilizantes/administração & dosagem , Dióxido de Silício/administração & dosagem , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Clorofilídeos , Feminino , Losartan/toxicidade , Masculino , Neoplasias Mamárias Experimentais/patologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos ICR , Porfirinas/toxicidade , Radiossensibilizantes/toxicidade , Dióxido de Silício/toxicidade , Carga Tumoral/efeitos dos fármacosRESUMO
Photodynamic therapy (PDT) has been applied in clinical cancer treatment. Here we report an aptamer-functionalized nanoscale metal-organic framework for targeted PDT. Our nanosystem can be easily prepared and successfully used for targeted PDT with a significantly enhanced therapeutic efficacy in vitro and in vivo. Methods: By combining the strong binding ability between phosphate-terminated aptamers and Zr-based nanoscale metal-organic frameworks (Zr-NMOFs) and the intercalation of photosensitizer TMPyP4 within the G-quadruplex DNA structure, TMPyP4-G4-aptamer-NMOFs were prepared. The characteristics and photodynamic performance of TMPyP4-G4-aptamer-NMOFs were examined after preparation. Then, we studied their stability, specific recognition ability, and phototoxicity in vitro. For in vivo experiments, the nanosystem was intratumorally injected into a HeLa subcutaneous xenograft tumor mouse model. After irradiation on day 0, mice were further injected with the nanosystem on day 5 and were again subjected to laser irradiation for 30 min. Tumor volumes and body weights of all mice were measured by caliper every 2 days after the treatment. Results: The nanosystem induced 90% cell death of targeted cells. In contrast, the control cells maintained about 40% cell viability at the same concentration of nanosystem. For the in vivo experiments, the nanosystem-treated group maintained more than 76% inhibition within the entire experimental period. Conclusion: We have demonstrated that our smart TMPyP4-G4-sgc8-NMOFs nanosystem can be used for targeted cancer therapy with high efficiency.
Assuntos
Aptâmeros de Nucleotídeos/administração & dosagem , Quadruplex G , Estruturas Metalorgânicas/administração & dosagem , Terapia de Alvo Molecular/métodos , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/administração & dosagem , Porfirinas/administração & dosagem , Animais , Aptâmeros de Nucleotídeos/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Portadores de Fármacos/administração & dosagem , Estabilidade de Medicamentos , Células HeLa , Xenoenxertos , Humanos , Estruturas Metalorgânicas/toxicidade , Camundongos , Transplante de Neoplasias , Neoplasias Experimentais/patologia , Neoplasias Experimentais/terapia , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/toxicidade , Resultado do TratamentoRESUMO
Ovarian Cancer is one of the deadliest gynecological cancer showing high resistance to chemotherapy. Non-overlapping and synergistic combination therapies are the best option to overcome this multi-pathological silent disease. Cationic peptides (CPs) with high targeting feature and ability to pass through cell membrane induce apoptosis via disruption of cancer cell membrane. Photodynamic Therapy (PDT) is a noninvasive clinically approved treatment modality combining light activated photosensitizer, light and oxygen. In this study we present, combination therapy composed of 9-mer +4 charge bearing CP and Benzoporphyrin derivative monoacid, (BPD-MA, Verteporfin) mediated PDT. In order to evaluate the effect of sequence on the outcome of the therapy, CP and BPD-MA mediated PDT was applied in two different sequence: 'CP first' 'BPD-MA first'. Treatment efficacy of combination therapy in SKOV-3 ovarian cancer cell line has been evaluated based on cell inhibition, cell death pathway, Combination index (CI), and Dose Reduction Index (DRI) values. When SKOV-3 ovarian cancer cell line treated with BPD-MA mediated PDT (5â¯J/cm2) and CP individually, IC30 values for each drug were determined as 1.1⯵M and 240⯵M respectively and apoptosis was the major death cell pathway for both of the drugs. In the case of combination therapy, SKOV-3 cell line treated with drugs in constant ratio yet on different sequence. Drugs were used in constant ratio so that one of them would not de-emphasize the effect of other in any concentration point. Our theoretical and experimental results were in agreement and showed that the treatment outcome significantly depends on the order of the treatment. For instance, while BPD-MA mediated PDT was applied prior to CP, cell inhibition at IC30 value of BPD-MA was roughly 28% with CI =3.3 suggesting antagonistic interaction between each therapy. When the sequence of treatment was changed to CP first, cell inhibition at IC30 concentration of CP was determined as 98% with CIâ¯=â¯0.3 creating substantial synergism between the drugs. Moreover, synergistic interactions were observed at all concentration points at CP first scenario. DRI value for CP first treatment option was much higher compared to BPD-MA first treatment making the former treatment sequence more attractive option for clinically relevant combination therapies. Based on our results, we strongly believe that 9-mer CP and BPD-MA-PDT based combination therapy, offering synergistic therapeutic outcome, may increase chances of treatment of ovarian cancer in comparison to 9-mer CP and/or BPD-MA alone case.
Assuntos
Apoptose/efeitos dos fármacos , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/toxicidade , Sequência de Aminoácidos , Apoptose/efeitos da radiação , Cátions/química , Linhagem Celular Tumoral , Sinergismo Farmacológico , Feminino , Humanos , Luz , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Peptídeos/química , Fármacos Fotossensibilizantes/química , Porfirinas/química , VerteporfinaRESUMO
BACKGROUND: Photodynamic therapy (PDT) of malignant brain tumors is a promising adjunct to standard treatment, especially if tumor stem cells thought to be responsible for tumor progression and therapy resistance were also susceptible to this kind of treatment. However, some photosensitizers have been reported to be substrates of ABCG2, one of the membrane transporters mediating resistance to chemotherapy. Here we investigate, whether inhibition of ABCG2 can restore sensitivity to photosensitizer chlorin e6-mediated PDT. METHODS: Accumulation of chlorin e6 in wild type U87 and doxycycline-inducible U251 glioblastoma cells with or without induction of ABCG2 expression or ABCG2 inhibition by KO143 was analyzed using flow cytometry. In U251 cells, ABCG2 was inducible by doxycycline after stable transfection with a tet-on expression plasmid. Tumor sphere cultivation under low attachment conditions was used to enrich for cells with stem cell-like properties. PDT was done on monolayer cell cultures by irradiation with laser light at 665nm. RESULTS: Elevated levels of ABCG2 in U87 cells grown as tumor spheres or in U251 cells after ABCG2 induction led to a 6-fold lower accumulation of chlorin e6 and the light dose needed to reduce cell viability by 50% (LD50) was 2.5 to 4-fold higher. Both accumulation and PDT response can be restored by KO143, an efficient non-toxic inhibitor of ABCG2. CONCLUSION: Glioblastoma stem cells might escape phototoxic destruction by ABCG2-mediated reduction of photosensitizer accumulation. Inhibition of ABCG2 during photosensitizer accumulation and irradiation promises to restore full susceptibility of this crucial tumor cell population to photodynamic treatment.
Assuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Dicetopiperazinas/toxicidade , Expressão Gênica/efeitos dos fármacos , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Fármacos Fotossensibilizantes/metabolismo , Porfirinas/metabolismo , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Clorofilídeos , Doxiciclina/farmacologia , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Luz , Niacinamida/análogos & derivados , Niacinamida/farmacologia , Compostos de Fenilureia/farmacologia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/química , Porfirinas/toxicidade , SorafenibeRESUMO
Photodynamic inactivation (PDI) of bacteria may play a major role in facing the challenge of the ever expanding antibiotic resistances. Here we report about the direct correlation of singlet oxygen luminescence kinetics and phototoxicity in E. coli cell suspension under PDI using the widely applied cationic photosensitizer TMPyP. Through direct access to the microenvironment, the time resolved investigation of singlet oxygen luminescence plays a key role in understanding the photosensitization mechanism and inactivation pathway. Using the homemade set-up for highly sensitive time resolved singlet oxygen luminescence detection, we show that the cationic TMPyP is localized predominantly outside the bacterial cells but in their immediate vicinity prior to photodynamic inactivation. Throughout following light exposure, a clear change in singlet oxygen kinetics indicates a redistribution of photosensitizer molecules to at least one additional microenvironment. We found the signal kinetics mirrored in cell viability measurements of equally treated samples from same overnight cultures conducted in parallel: A significant drop in cell viability of the illuminated samples and stationary viability of dark controls. Thus, for the system investigated in this work - a Gram-negative model bacteria and a well-known PS for its PDI - singlet oxygen kinetics correlates with phototoxicity. This finding suggests that it is well possible to evaluate PDI efficiency directly via time resolved singlet oxygen detection.
Assuntos
Escherichia coli/efeitos dos fármacos , Fármacos Fotossensibilizantes/toxicidade , Oxigênio Singlete/metabolismo , Escherichia coli/metabolismo , Escherichia coli/efeitos da radiação , Cinética , Luz , Fármacos Fotossensibilizantes/química , Porfirinas/química , Porfirinas/toxicidade , Oxigênio Singlete/química , Água/químicaRESUMO
Amphiphilic porphyrin photosensitisers (PSs) having combinations of directly substituted pyridyl group(s) at the meso-position of a porphyrin macrocycle, and/or indirectly linked pyridyl groups as benzamide derivatives are reported. The compounds 5,10,15,20-tetrakis-(4-pyridylbenzamide)porphyrin (A.2), 5,10,15,20-tetra[N-(pyridine-4-yl)benzamidium] porphyrin (A.3), 5-mono-(4-pyridyl)-10,15,20-tris-(4-pyridylbenzamide)porphyrin (B.2) and 5-mono-(4-methylpyridinium)-10,15,20-tris-(4-pyridiniumbenzamide)porphyrin (B.3) were synthesised. The compounds were successfully characterised through UV-Vis, Emission, 1H NMR, and ESI-HRMS techniques. To evaluate the effect of this combination of directly conjugated and non-conjugated pyridyl/cationic pyridinium groups on the porphyrin macrocycle, the efficacy of the synthesised compounds was compared to a known standard 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin (TMPyP). These compounds show better efficacy (IC50's ranging between 0.66±0.04µM to 3.71±1.01µM) against A549 (human epithelial adenocarcinoma lung cancer) cell line under in vitro photodynamic conditions in comparison to MDA-MB-231 (breast cancer) (IC50's ranging between 3.7±0.087µM to 12.1±0.12µM) and Pa-1 (ovarian cancer) (IC50's ranging between 17.9±0.01µM to 42.45±0.02µM) cell lines. It was found that B.3, having a pyridinium group attached to the meso-position of the macrocycle along with three distal cationic pyridinium groups, independent of the porphyrinic electron delocalisation cycle, showed better photocytotoxic efficacy (IC50=0.66±0.04µM, A549 lung cancer cell line) and higher potential to promote apoptosis and hence better efficacy as PS towards cancer photodynamic therapy (PDT). The PDT activity of B.3 was further verified and established by various biological assays, viz. Annexin V assay, cell cycle assay, and reactive oxygen species (ROS) activity assay.
Assuntos
Benzamidas/química , Fármacos Fotossensibilizantes/química , Porfirinas/química , Células A549 , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Luz , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/síntese química , Porfirinas/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Espectrofotometria , Espectrofotometria UltravioletaRESUMO
Photodynamic therapy is selective destruction of cells stained with a photosensitizer upon irradiation with light at a specific wavelength in the presence of oxygen. Cell death upon photodynamic treatment is known to occur mainly due to free radical production and subsequent development of oxidative stress. During photodynamic therapy of brain tumors, healthy cells are also damaged; considering this, it is important to investigate the effect of the treatment on normal neurons and glia. We employed live-cell imaging technique to investigate the cellular mechanism of photodynamic action of radachlorin (200 nM) on neurons and astrocytes in primary rat cell culture. We found that the photodynamic effect of radachlorin increases production of reactive oxygen species measured by dihydroethidium and significantly decrease mitochondrial membrane potential. Mitochondrial depolarization was independent of opening of mitochondrial permeability transition pore and was insensitive to blocker of this pore cyclosporine A. However, irradiation of cells with radachlorin dramatically decreased NADH autofluorescence and also reduced mitochondrial NADH pool suggesting inhibition of mitochondrial respiration by limitation of substrate. This effect could be prevented by inhibition of poly (ADP-ribose) polymerase (PARP) with DPQ. Thus, irradiation of neurons and astrocytes in the presence of radachlorin leads to activation of PARP and decrease in NADH that leads to mitochondrial dysfunction.
Assuntos
Astrócitos/efeitos da radiação , Lasers Semicondutores/efeitos adversos , Mitocôndrias/efeitos da radiação , Neurônios/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Fármacos Fotossensibilizantes/toxicidade , Animais , Astrócitos/metabolismo , Técnicas de Cocultura , Combinação de Medicamentos , Mitocôndrias/metabolismo , Neurônios/metabolismo , Estresse Oxidativo/fisiologia , Porfirinas/toxicidade , RatosRESUMO
The inhibitor of DNA binding and cell differentiation (Id) proteins are dominant negative regulators of the helix-loop-helix transcription factor family and play a key role during development as well as in vascular disorders and cancer. In fact, impairing the Id-protein activity in cancer cells reduces cell growth and even chemoresistance. Recently, we have shown that a synthetic Id-protein ligand (1Y) consisting of a cyclic nonapeptide can reduce the viability of the two breast cancer cell lines MCF-7 and T47D and of the bladder cancer cells T24 to about 50% at concentrations ≥100µM. Moreover, the cyclopeptide displays both proapoptotic and antiproliferative effects on MCF-7 cells. Herein, we show that the cyclopeptide does not induce cell death at the dose of 5µΜ, but it still inhibits MCF-7 and T24 cell proliferation, which correlates with an increased protein level of the cell-cycle regulator p27Kip1. Furthermore, 1Y-pretreated MCF-7, T47D, and T24 cells are more susceptible than untreated cells to the phototoxic effects of the three photosensitizers meta-tetra(hydroxyphenyl)chlorin, porfimer sodium, and hypericin, which are applied in photodynamic therapy (PDT). The combination of the Id-protein ligand with each of the light-activated photosensitizers shows synergistic effects on the reduction of cell viability. In conclusion, an Id-protein ligand with moderate cancer cell killing activity at concentrations ≥100µM can be applied at a 20-fold lower and barely toxic dose to raise the sensitivity of cancer cells towards phototoxicity associated with photodynamic treatment. This suggests the potential benefit of targeting the Id proteins in combined drug approaches for cancer therapy.
Assuntos
Proliferação de Células/efeitos dos fármacos , Proteína 1 Inibidora de Diferenciação/antagonistas & inibidores , Peptídeos Cíclicos/toxicidade , Fármacos Fotossensibilizantes/toxicidade , Antracenos , Linhagem Celular Tumoral , Proliferação de Células/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Éter de Diematoporfirina/toxicidade , Sinergismo Farmacológico , Humanos , Proteína 1 Inibidora de Diferenciação/metabolismo , Luz , Células MCF-7 , Nanoestruturas/química , Nanoestruturas/toxicidade , Peptídeos Cíclicos/química , Perileno/análogos & derivados , Perileno/toxicidade , Fármacos Fotossensibilizantes/química , Porfirinas/toxicidadeRESUMO
A treatment of onychomycosis using the photodynamic effect would be a favorable alternative to currently used antimycotic drugs. This study should be considered as a first step towards development and control of an efficient photodynamic inactivation of onychomycosis causative pathogens. Here, we evaluate the usage of time-resolved 2D singlet oxygen luminescence detection in combination with 2D fluorescence scanning as a tool to understand the behavior of the photosensitizer when applied to fungi on Petri dishes. To investigate the interaction of photosensitizer with fungi in various concentrations and in different stages of live, a photodynamic inactivation was avoided by keeping the samples in darkness. Scans of singlet oxygen luminescence and photosensitizer fluorescence were performed over a period of 24days. Two different photosensitizer, a cationic porphyrin and cationic corrole and two fungi strains, the dermatophyte Trichophyton rubrum and the mold Scopulariopsis brevicaulis, were investigated in this study. The two-dimensional correlation of photosensitizer fluorescence and singlet oxygen luminescence revealed differences in the diffusion of both photosensitizer. Even though the singlet oxygen luminescence was quenched with increasing growth of fungi, it was found that the kinetics of singlet oxygen luminescence could be detected on Petri dishes for both photosensitizers and both fungi strains for up to seven days.
Assuntos
Medições Luminescentes , Fármacos Fotossensibilizantes/química , Oxigênio Singlete/química , Cinética , Luz , Fármacos Fotossensibilizantes/toxicidade , Porfirinas/química , Porfirinas/toxicidade , Scopulariopsis/efeitos dos fármacos , Scopulariopsis/efeitos da radiação , Oxigênio Singlete/metabolismo , Trichophyton/efeitos dos fármacos , Trichophyton/efeitos da radiaçãoRESUMO
DNA polymorphism exerts a fascination on a large scientific community. Without crystallographic structural data, clarification of the binding modes between G-quadruplex (G4) and ligand (complex) is a challenging job. In the present work, three porphyrin compounds with different flexible carbon chains (arms) were designed, synthesized and characterized. Their binding, folding and stabilizing abilities to human telomeric G4 DNA structures were comparatively researched. Positive charges at the end of the flexible carbon chains seem to be favorable for the DNA-porphyrin interactions, which were evidenced by the spectral results and further confirmed by the molecular docking calculations. Biological function analysis demonstrated that these porphyrins show no substantial inhibition to Hela, A549 and BEL 7402 cancer cell lines under dark while exhibit broad inhibition under visible light. This significantly enhanced photocytotoxicity relative to the dark control is an essential property of photochemotherapeutic agents. The feature of the flexible arms emerges as critical influencing factors in the cell photocytotoxicity. Moreover, an ROS-mediated mitochondrial dysfunction pathway was suggested for the cell apoptosis induced by these flexible-armed porphyrins. It is found that the porphyrins with positive charges located at the end of the flexible arms represent an exciting opportunity for photochemotherapeutic anti-cancer drug design.