RESUMO
Controlled photoreduction of Pt(IV) prodrugs is a challenging task due to the possibility of targeted light-controlled activation of anticancer agents without affecting healthy tissues. Also, a conjugation of photosensitizers and clinically used platinum drugs into one Pt(IV) prodrug allows combining photodynamic therapy and chemotherapy approaches into one molecule. Herein, we designed the cisplatin-based Pt(IV) prodrug Riboplatin with tetraacetylriboflavin in the axial position. A novel Pt(IV) prodrug is able to act both as a photodynamic therapy (PDT) agent through the conversion of ground-state 3O2 to excited-state 1O2 and as an agent of photoactivated chemotherapy (PACT) through releasing of cisplatin under gentle blue light irradiation, without the requirement of a reducing agent. The light-induced behavior of Riboplatin was investigated using an electrochemical sensor in MCF-7 tumor spheroids. Photocontrolled cisplatin release and ROS generation were detected electrochemically in real time. This appears to be the first confirmation of simultaneous photoactivated release of anticancer drug cisplatin and ROS from a dual-action Pt(IV) prodrug observed from the inside of living tumor spheroids.
Assuntos
Antineoplásicos , Pró-Fármacos , Cisplatino/farmacologia , Cisplatino/química , Pró-Fármacos/farmacologia , Pró-Fármacos/química , Espécies Reativas de Oxigênio , Antineoplásicos/farmacologia , Antineoplásicos/química , Platina/química , Linhagem Celular TumoralRESUMO
We report herein a Pt(IV) prodrug with metronidazole in axial positions Pt-Mnz. The nitroaromatic axial ligand was conjugated with a cisplatin scaffold to irreversibly reduce under hypoxic conditions, thereby retaining the Pt(IV) prodrug in the area of hypoxia. X-ray near-edge adsorption spectroscopy (XANES) on dried drug-preincubated tumor cell samples revealed a gradual release of cisplatin from the Pt-Mnz prodrug instead of rapid intracellular degradation. The ability of the prodrug to penetrate into three-dimensional (3D) spheroid cellular cultures was evaluated by a novel electrochemical assay via a platinum-coated carbon nanoelectrode, capable of single-cell measurements. Using a unique technique of electrochemical measurements in single tumor spheroids, we were able to both detect the real-time response of the axial ligand to hypoxia and establish the depth of penetration of the drug into the tumor model.
Assuntos
Antineoplásicos , Pró-Fármacos , Antineoplásicos/química , Antineoplásicos/farmacologia , Carbono , Linhagem Celular Tumoral , Cisplatino/química , Humanos , Hipóxia , Ligantes , Metronidazol/farmacologia , Platina/química , Pró-Fármacos/química , Pró-Fármacos/farmacologiaRESUMO
We report herein the design, synthesis, and biological investigation of a series of novel Pt(IV) prodrugs with non-steroidal anti-inflammatory drugs naproxen, diclofenac, and flurbiprofen, as well as these with stearic acid in the axial position. Six Pt(IV) prodrugs 5-10 were designed, which showed superior antiproliferative activity compared to cisplatin as well as an ability to overcome tumor cell line resistance to cisplatin. By tuning the drug lipophilicity via variation of the axial ligands, the most potent Pt(IV) prodrug 7 was obtained, with an enhanced cellular accumulation of up to 153-fold that of cisplatin and nanomolar cytotoxicity both in 2D and 3D cell cultures. Pt2+ species were detected at different depths of MCF-7 spheroids after incubation with Pt(IV) prodrugs using a Pt-coated carbon nanoelectrode. Cisplatin accumulation in vivo in the murine mammary EMT6 tumor tissue of BALB/c mice after Pt(IV) prodrug injection was proved electrochemically as well. The drug tolerance study on BALB/c mice showed good tolerance of 7 in doses up to 8 mg/kg.
Assuntos
Anti-Inflamatórios não Esteroides , Antineoplásicos , Compostos de Platina , Pró-Fármacos , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Cisplatino/farmacologia , Desenho de Fármacos , Humanos , Células MCF-7 , Camundongos , Camundongos Endogâmicos BALB C , Compostos de Platina/farmacologia , Pró-Fármacos/farmacologiaRESUMO
A series of 73 ligands and 73 of their Cu+2 and Cu+1 copper complexes with different geometries, oxidation states of the metal, and redox activities were synthesized and characterized. The aim of the study was to establish the structure-activity relationship within a series of analogues with different substituents at the N(3) position, which govern the redox potentials of the Cu+2/Cu+1 redox couples, ROS generation ability, and intracellular accumulation. Possible cytotoxicity mechanisms, such as DNA damage, DNA intercalation, telomerase inhibition, and apoptosis induction, have been investigated. ROS formation in MCF-7 cells and three-dimensional (3D) spheroids was proven using the Pt-nanoelectrode. Drug accumulation and ROS formation at 40-60 µm spheroid depths were found to be the key factors for the drug efficacy in the 3D tumor model, governed by the Cu+2/Cu+1 redox potential. A nontoxic in vivo single-dose evaluation for two binuclear mixed-valence Cu+1/Cu+2 redox-active coordination compounds, 72k and 61k, was conducted.
Assuntos
Complexos de Coordenação/química , Cobre/química , Imidazóis/química , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Complexos de Coordenação/metabolismo , Complexos de Coordenação/farmacologia , Cristalografia por Raios X , Dano ao DNA/efeitos dos fármacos , Humanos , Ligantes , Células MCF-7 , Modelos Biológicos , Conformação Molecular , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Esferoides Celulares/efeitos dos fármacos , Relação Estrutura-Atividade , Telomerase/antagonistas & inibidores , Telomerase/metabolismoRESUMO
Current in vitro and animal tests for drug development are failing to emulate the systemic organ complexity of the human body and, therefore, often do not accurately predict drug toxicity, leading to high attrition rates in clinical studies (Paul et al., 2010). The phylogenetic distance between humans and laboratory animals is enormous, this affects the transferability of animal data on the efficacy of neuroprotective drugs. Therefore, many neuroprotective treatments that have shown promise in animals have not been successful when transferred to humans (Dragunow, 2008; Gibbons and Dragunow, 2010). We present a multi-organ chip capable of maintaining 3D tissues derived from various cell sources in a combined media circuit which bridges the gap in systemic and human tests. A steady state co-culture of human artificial liver microtissues and human neurospheres exposed to fluid flow over two weeks in the multi-organ chip has successfully proven its long-term performance. Daily lactate dehydrogenase activity measurements of the medium and immunofluorescence end-point staining proved the viability of the tissues and the maintenance of differentiated cellular phenotypes. Moreover, the lactate production and glucose consumption values of the tissues cultured indicated that a stable steady-state was achieved after 6 days of co-cultivation. The neurospheres remained differentiated neurons over the two-week cultivation in the multi-organ chip, proven by qPCR and immunofluorescence of the neuronal markers ßIII-tubulin and microtubule-associated protein-2. Additionally, a two-week toxicity assay with a repeated substance exposure to the neurotoxic 2,5-hexanedione in two different concentrations induced high apoptosis within the neurospheres and liver microtissues, as shown by a strong increase of lactate dehydrogenase activity in the medium. The principal finding of the exposure of the co-culture to 2,5-hexanedione was that not only toxicity profiles of two different doses could be discriminated, but also that the co-cultures were more sensitive to the substance compared to respective single-tissue cultures in the multi-organ-chip. Thus, we provide here a new in vitro tool which might be utilized to predict the safety and efficacy of substances in clinical studies more accurately in the future.
Assuntos
Técnicas de Cocultura/métodos , Fígado/citologia , Técnicas Analíticas Microfluídicas/métodos , Neurônios/metabolismo , Esferoides Celulares/citologia , Diferenciação Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Hexanonas/toxicidade , Humanos , Fígado/efeitos dos fármacos , Esferoides Celulares/efeitos dos fármacos , Técnicas de Cultura de Tecidos/métodosRESUMO
Exercise can alter human health in both beneficial (e. g. reduced risk of infection and of atherosclerosis) and adverse (e. g. anaphylaxis, exercise-induced asthma, and exacerbation of chronic illness) ways. Hitherto, the mechanisms linking exercise and health are not fully understood, but may rest on the capability of exercise to both increase circulating immune cells and modulate their activity. Natural killer (NK) cells, a major component of innate immunity, are one of the most sensitive populations of immune cells to exercise stress. NK cells play an important role in the detection and elimination of tumours and virus-infected cells. To mediate NK cell functions, there is an array of activating and inhibitory receptors with distinct specificities on their surface. Killer-cell immunoglobulin-like receptors (KIRs) which bind to MHC class I are a key example of receptors expressed by NK cells. The combination of MHC class I and KIR variants influences resistance to infections, susceptibility to autoimmune diseases, as well as complications of pregnancy. It is suggested that KIRs may also determine a considerable part of the effects of physical activity on human health. In this review we discuss KIRs in more detail, their role in the onset of human diseases, and the influence of acute exercise on KIR gene expression.