RESUMO
This paper describes the synthesis of trans-bis-(3-aminoflavone)dichloridoplatinum(ii) (trans-Pt(3-af)2Cl2; TCAP) for use as a potential anticancer compound, and the evaluation of its structure by elemental and spectral analyses, and X-ray crystallography. The complex demonstrated a significant cytotoxic effect against human and murine cancer cell lines, as well as weaker toxicity towards healthy cells (human peripheral blood lymphocytes) in comparison with cisplatin. Various biochemical and morphological methods confirm that the proapoptotic activity of trans-Pt(3-af)2Cl2 is markedly higher than the reference cisplatin. Our results suggest that trans-Pt(3-af)2Cl2 may have a different antitumour specificity from that of cisplatin.
Assuntos
Complexos de Coordenação/química , Flavonoides/química , Platina/química , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Linhagem Celular , Complexos de Coordenação/síntese química , Complexos de Coordenação/toxicidade , Cristalografia por Raios X , Fragmentação do DNA/efeitos dos fármacos , Humanos , Isomerismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Microscopia de Fluorescência , Conformação MolecularRESUMO
In this study we examined their proapoptotic activity of cis-dichloridobis(3-imino-2-methoxyflavanone)ruthenium(II)3H(2)O (1) and cis-dichloridobis(3-imino-2-ethoxyflavanone)ruthenium(II)2H(2)O (2) towards human bladder carcinoma cell line EJ and its cisplatin resistant subline EJcisR. On the basis of the experiments we carried out, it may be concluded, that: CDDP (cis-diamminedichloridoplatinum) resistance of EJcisR cells is probably based on partial loss of apoptotic pathway activating caspase-8 and increased resistance to DNA strand breaks and/or alkali-labile sites. Increased glutathione levels, as well as activity of P-gp transporter seems to be not relevant in this case. The proapoptotic activity of the ruthenium compounds is higher than that of cisplatin. Higher proapoptotic activity of 1 and 2 when compared to CDDP may be due to the presence of large, lipophilic flavanone-based ligands that may facilitate their trans-membrane transport and their redox activity. 1 and 2 induce apoptosis apparently in more than one way. Although caspase-8 activation and DNA strand breaks and/or alkali-labile sites are caused by the compounds, their ability to cause the oxidative stress in the cells may also participate in apoptosis induction.
Assuntos
Antineoplásicos/farmacologia , Apoptose , Cisplatino/farmacologia , Complexos de Coordenação/farmacologia , Carcinoma/tratamento farmacológico , Carcinoma/metabolismo , Caspase 8/metabolismo , Complexos de Coordenação/química , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Flavanonas/química , Humanos , Ligantes , Estresse Oxidativo , Rodamina 123/farmacologia , Compostos de Rutênio/química , Compostos de Rutênio/farmacologia , Células Tumorais Cultivadas , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/metabolismoRESUMO
Synthesis, structure and properties of two new flavanone complexes of Ru(ii) are described. The new complexes form during the reaction of ruthenium(iii) chloride with 3-aminoflavone (3-af) dissolved in an aliphatic alcohol. The formed products depend on the alcohol used and were found to be: cis-dichloridobis(3-imino-2-methoxyflavanone)ruthenium(ii)·3H(2)O (1) from a methanolic solution and cis-dichloridobis(3-imino-2-ethoxyflavanone)ruthenium(ii)·2H(2)O (2) from an ethanolic solution, in which the original ligand 3-af had been converted by dehydrogenative alcoholysis to an entirely new ligand. This paper presents the X-ray structure and detailed (1)H-NMR analysis of both new compounds, as well as the study of their antiproliferative activity. The coordination of Ru(ii) is octahedral with [RuCl(2)N(2)O(2)] chromophores, having trans chlorides and common Ru-L distances. Both 1 and 2 are highly cytotoxic towards the cisplatin resistant EJ and L1210 cell lines, and both complexes are as active as cisplatin in the sensitive cell lines. They display the ability to overcome cisplatin resistance in the drug resistant sub-lines EJcisR and L1210R. The present evidence suggests that the mechanism of biological activity may be different for these ruthenium compounds compared to cisplatin.
Assuntos
Complexos de Coordenação/química , Flavonoides/química , Rutênio/química , Animais , Linhagem Celular Tumoral , Complexos de Coordenação/síntese química , Complexos de Coordenação/toxicidade , Cristalografia por Raios X , Humanos , Ligação de Hidrogênio , Ligantes , Espectroscopia de Ressonância Magnética , Camundongos , Conformação MolecularRESUMO
cis-DDP (cis-diamminedichloroplatinum(II), CAS 15663-27-1) is widely used in chemotherapy of many types of cancer. However, besides effectiveness, it gives many side effects which limit its clinical application. Therefore, nowadays studies are focused on searching for novel analogs of cis-DDP, at least equally effective in chemotherapy but less toxic. One of them might be cis-BAFDP (cis-bis(3-aminoflavone)dichloroplatinum(II)) with one of the hydrogen atoms of the amino group of cis-DDP replaced by a flavone ring. 3-Aminoflavone (AF) which posseses the desired NH2 group has been used as non-leaving ligand. The complex has been obtained in the reaction of AF and K2PtCl4. There are no data concerning evaluation of structural studies of cis-BAFDP, the beneficial anticancer properties of which were proved in vitro and in vivo. Therefore it was worthwhile to undertake a confirmation of the chemical structure of this compound by applying various spectroscopic techniques especially because of its potential pharmacological application. With this aim in mind this compound was characterized by: IR, 1H NMR, 195Pt NMR, UV absorption and fluorescence spectroscopy. Moreover, stronger apoptosis induction by cis-BAFDP than cis-DDP in the human nonsmall cancer cell line A549 was observed using Hoechst 33258/propidium iodide double staining.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Compostos Organoplatínicos/síntese química , Compostos Organoplatínicos/farmacologia , Bisbenzimidazol , Linhagem Celular Tumoral , Corantes , Humanos , Espectroscopia de Ressonância Magnética , Necrose , Espectrometria de Fluorescência , Espectrofotometria Infravermelho , Espectrofotometria Ultravioleta , Relação Estrutura-AtividadeRESUMO
Currently cis-diamminedichloroplatinum(II) (cis-DDP) is one of the most commonly applied compounds in chemotherapy of many types of cancer. However, a drawback is that its effectiveness presents with many side effects. Therefore, human normal lymphocytes were chosen as a model system to study cis-bis(3-aminoflavone)dichloroplatinum(II) (the cis-Pt(II) complex of 3-aminoflavone) in comparison with cis-DDP. We examined the effect of both tested compounds on cell viability and induction of apoptosis and necrosis. Trypan blue and acridine orange/ethidium bromide staining were carried out, as well as quantitative analysis of the apoptotic signal of P53 and BAX induction caused by the cis-Pt(II) complex of 3-aminoflavone in comparison with cis-DDP. cis-DDP induced a decrease of cell viability and led to a higher increase in necrosis and apoptosis than did the cis-Pt(II) complex of 3-aminoflavone. Moreover, at the molecular level cis-DDP increased P53 and BAX expression in comparison with the other tested compound. The cis-Pt(II) complex of 3-aminoflavone showed a weaker genotoxic effect in normal lymphocytes in comparison with cis-DDP, which was a stronger inducer of apoptosis and necrosis.
Assuntos
Apoptose/efeitos dos fármacos , Cisplatino/toxicidade , Flavonoides/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Genes p53/efeitos dos fármacos , Linfócitos/citologia , Proteína X Associada a bcl-2/genética , Sequência de Bases , Técnicas de Cultura de Células , Sobrevivência Celular/efeitos dos fármacos , Primers do DNA , Humanos , Linfócitos/efeitos dos fármacos , Linfócitos/fisiologia , Necrose , RNA Mensageiro/genética , Proteína X Associada a bcl-2/efeitos dos fármacosRESUMO
Lung cancer remains one of the most common causes of cancer-related death worldwide. Approximately 80% is histologically non-small cell lung carcinoma (NSCLC) and in about 70% of patients it is an unresectable type. Clinical studies indicated that application of platinum derivatives caused good results and combinations of platinum with other agents could improve median survivals. In view of the central problem of sufficient efficiency of drugs in chemotherapy, efforts have focused on the development of alternative platinum-based analogues that can be more effective in cancer treatment. cis-bis(3-aminoflavone)dichloroplatinum(II) (cis-Pt(II) complex of 3-aminoflavone) represents a novel class of platinum-based potential antitumour agents. In order to evaluate the degree of apoptosis, acridine orange/ethidium bromide and Hoechst 33258/propidum iodide double staining as well as RT-PCR (P53 and BAX expression evaluation) were used in lung cancer cell line A549 after treatment with this compound in comparison with cis-diamminedichloroplatinum(II) (cis-DDP). Apoptotic cells at early and late stages and also necrotic ones were observed after usage of cis-Pt(II) complex of 3-aminoflavone and the percentage of these cells outnumbered the values obtained after cis-DDP application. The former compound induced a higher percentage of P53 and BAX expression in A549 cells in comparison with the latter one. Results indicate the beneficial properties of cis-Pt(II) complex of 3-aminoflavone as a potential antitumor drug.
Assuntos
Antineoplásicos/farmacologia , Cisplatino/farmacologia , Compostos Organoplatínicos/farmacologia , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismo , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , RNA Mensageiro/metabolismo , Fatores de Tempo , Proteína Supressora de Tumor p53/biossíntese , Proteína Supressora de Tumor p53/genética , Proteína X Associada a bcl-2/biossíntese , Proteína X Associada a bcl-2/genéticaRESUMO
The interaction of cis-diamminedichloroplatinum(II) (cis-DDP) and the potential novel chemotherapeutic agent, cis-bis(3-aminoflavone)dichloroplatinum(II) (cis-BAFDP) was studied electrochemically with calf thymus double-stranded DNA (dsDNA) by using differential pulse voltammetry (DPV) with disposable pencil graphite electrode (PGE) at the surface. These studies were prompted by beneficial biological properties of cis-BAFDP in comparison with cis-DDP, which were proven in vitro both in human normal and cancer cells and in vivo. The changes in the experimental parameters such as the concentration of cis-DDP and cis-BAFDP were studied by using DPV; in addition, the reproducibility of this genosensor and the detection limit for each compound were determined. After the interaction of cis-DDP with dsDNA, the DPV signal of guanine and adenine was found to be decreasing. In comparison with cis-DDP, a dramatic decrease at adenine signal was also obtained after the interaction of cis-BAFDP and dsDNA. Similar results were also found in solution phase after the latter compound interacts with poly[A]. The features of the proposed electrochemical method for the detection of cis-BAFDP with DNA in comparison with cis-DDP are discussed and compared with those methods previously reported for the other type of DNA-targeted agents in the literature.
Assuntos
Antineoplásicos/farmacologia , Cisplatino/farmacologia , DNA/efeitos dos fármacos , DNA/genética , Compostos Organoplatínicos/farmacologia , Adenina/química , Eletroquímica , Eletrodos , Grafite , Oxirredução , Tensoativos , Sais de Tetrazólio , TiazóisRESUMO
Non-small cell lung cancer (NSCLC) includes a group of tumors that respond poorly to drugs. cis-Diamminedichloroplatinum(II) (cis-DDP) toxicity still remains problematic, and not completely solved by the improvement of supportive care. Therefore, the cis-Pt(II) complex of 3-aminoflavone was selected from cis-DDP analogues with a more favourable toxic profile towards normal cells and at least similar or better anti-tumor activity in comparison with cis-DDP. The aim of this research is to compare the ability of the cis-Pt(II) complex of 3-aminoflavone and cis-DDP to induce apoptosis and necrosis in the human non-small cancer cell line A549. Trypan blue dye exclusion, fluorochrome staining (acridine orange/ethidium bromide double staining), MTT and TUNEL (TdT-mediated dUTP Nick-End Labeling) assays were used. The results obtained show that the cis-Pt(II) complex of 3-aminoflavone is more active in inducing apoptosis and necrosis and in decreasing viability in A549 cells than cis-DDP, which suggests that it could be a potential chemotherapeutic drug.
Assuntos
Antineoplásicos/toxicidade , Apoptose , Cisplatino/toxicidade , Flavonoides/metabolismo , Necrose , Laranja de Acridina/metabolismo , Antineoplásicos/química , Divisão Celular , Linhagem Celular Tumoral , Sobrevivência Celular , Cisplatino/química , Cisplatino/metabolismo , Etídio/metabolismo , Flavonoides/química , Corantes Fluorescentes/metabolismo , Humanos , Marcação In Situ das Extremidades Cortadas , Estrutura Molecular , Sais de Tetrazólio/metabolismo , Tiazóis/metabolismoRESUMO
Cis-diamminedichloroplatinum(II) (cis-DDP) is one of the most successful antineoplastic drugs. However, besides effectiveness it gives many side effects. Therefore, current studies are concentrated on searching for new analogs equally effective in chemotherapy but less toxic. Comparison of genotoxic properties of cis-Pt(II) complex of 3-aminoflavone and cis-DDP in a comet assay with and without H2O2 application was performed in A549 cell line. The higher tail moment values were noticed for the former compound contrary to the latter one in both variants. It suggests mainly DNA breaks (besides cross-links) appearance after cis-Pt(II) complex of 3-aminoflavone application and might indicate DNA degradation in comparison with cis-DDP.
Assuntos
Cisplatino/análogos & derivados , Cisplatino/toxicidade , Ensaio Cometa/métodos , Flavonoides/toxicidade , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Flavonoides/química , HumanosRESUMO
Short-term tests that detect genetic damage have provided information needed for evaluating carcinogenic risks of chemicals to man. The mutagenicity of cis-bis(3-aminoflavone)dichloroplatinum(II) (cis-[Pt(AF)2Cl2]) in comparison with cis-diamminedichloroplatinum(II) (cis-DDP) was evaluated in the standard plate-incorporation assay in four strains of Salmonella typhimurium: TA97a, TA98, TA100 and TA102, in experiments with and without metabolic activation. It was shown that cis-[Pt(AF)2Cl2] acts directly and is mutagenic for three strains of S. typhimurium: TA97a, TA98 and TA100. In comparison with cis-DDP this compound showed a weaker genotoxicity. Contrary to cis-DDP it has not shown toxic properties in the tester bacteria. The genotoxicity of both tested compounds was evaluated using chromosomal aberration, sister chromatid exchange and micronucleus assays, without and with metabolic activation, in human lymphocytes in vitro. The inhibitory effects of both compounds on mitotic activity, cell proliferation kinetics and nuclear division index were also compared. In all test systems applied, cis-[Pt(AF)2Cl2] was a less effective clastogen and a weaker inducer of both sister chromatid exchanges and micronuclei in comparison with cis-DDP, with and without metabolic activation. cis-[Pt(AF)2Cl2] has a direct mechanism of action and is less cytostatic and cytotoxic than the other compound. These results provide important data on the genotoxicity of cis-[Pt(AF)2Cl2] and indicate its beneficial properties as a potential anticancer drug, especially in comparison with cis-DDP.
Assuntos
Cisplatino/farmacologia , Compostos Organoplatínicos/toxicidade , Aberrações Cromossômicas , Humanos , Testes de Mutagenicidade , Salmonella typhimurium/efeitos dos fármacosRESUMO
cis-Diamminedichloroplatinum(II) (cis-DDP) is one of the most widely administrated antitumor drugs. However, the use of cis-DDP is severely limited because of its toxic side effects. Therefore, efforts are concentrated on the development of improved platinum compounds with a broader activity spectrum and effectiveness in chemotherapy, but lower toxicity. Beneficial properties of flavonoids, e.g. their antitumor activity, encouraged scientists to synthesize cis-bis(3-aminoflavone)dichloroplatinum(II). Abilities of these compounds to induce apoptosis and necrosis were compared by use of trypan blue, fluorochrome staining (Hoechst 33258/propidium iodide double staining) and TUNEL assays. The cytotoxicity was evaluated by MTT. The results obtained show that the cis-Pt(II) complex of 3-aminoflavone is less toxic than cis-DDP. However, the former compound has a faster rate of apoptosis induction in lymphocytes than the latter. The cis-Pt(II) complex of 3-aminoflavone induces apoptosis in normal lymphocytes to a lesser degree and could be a potential antitumor drug.