Avaliação toxicogenética e estudo dos mecanismos de resposta antiproliferativa de metalofármacos de rutênio contendo bioligantes naftoquinônicos em modelos celulares tumorais convencionais e em 3D
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Data
2020-01-29
Autores
De Grandis, Rone Aparecido [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Recentemente, complexos de rutênio têm ocupado lugar de destaque na busca por novas terapias anticâncer em razão de sua potencialidade de ação, seletividade em tecido tumoral e toxicidade inferior aos complexos de platina, atualmente utilizados na clínica. A diversidade de seus ligantes é fundamental para que esses metalofármacos atinjam múltiplos alvos biológicos e assim, superem as limitações das terapias existentes. Uma abordagem para moléculas multialvo é a combinação de ligantes bioativos com propriedades antitumorais comprovadas, como os derivados de naftoquinonas naturais. Baseados nisso, esse estudo propôs a avaliação e caracterização das propriedades toxicogenéticas e antitumorais de oito novos complexos de rutênio contendo os ligantes naturais lapachol e lausona. De maneira geral, a série de complexos fosfínicos apresentou satisfatória atividade antiproliferativa frente a diferentes linhagens celulares tumorais, em especial para células derivadas de carcinoma de próstata. Os valores de IC50 desses complexos foram consideravelmente menores do que os valores encontrados para a cisplatina, no qual foi observada atividade até 28 vezes mais seletiva, em relação a esse fármaco. Além disso, os ensaios de eficiência clonogênica sugerem que a substituição do ligante bipy pela phen aumenta a atividade dos compostos, induzindo a perda da clonogenicidade das células tumorais de próstata DU-145. Os complexos fosfínicos também apresentaram atividade antiproliferativa em modelos celulares em três dimensões (3D). Nesses ensaios, os complexos contendo o ligante lapachol demonstraram atividade citotóxica independente do estado de hipóxia encontrado no interior dos esferoides. Ademais, os resultados indicam que o complexo DP-LP é capaz de permear através do esferoide tumoral e desestabilizar as agregações celulares, resultando na perturbação das interações célula-matriz. De fato, o complexo DP-LP desencadeia apoptose em células DU-145 acompanhado de um acúmulo de células na fase G2/M, o que é consistente com a inibição da atividade da proteína AURORA-B. O mecanismo central relacionado com esses efeitos é de que a presença do lapachol no complexo DP-LP promova a geração de espécies reativas intracelulares que podem causar danos ao DNA. A fragmentação do DNA, evidenciada pelo ensaio do cometa pode ativar sistemas de reparos de DNA que mantém as células no checkpoint G2/M devido a diminuição da expressão do gene AURKB. Além disso, os danos ao DNA configuram o estímulo que desencadeia a ativação de caspases que conduz as células DU-145 à apoptose. Em suma, os resultados encontrados neste estudo contribuem para a seleção e elucidação das propriedades antiproliferativas e citotóxicas de novos compostos, demonstrando que a coordenação do lapachol em complexos fosfínicos de rutênio(II) possibilita a formação de complexos com potencial de impacto na área de saúde tendo como alvo o tratamento do câncer.
Recently, ruthenium complexes have occupied a leading position in the search for new anti-cancer therapies due to their potential of action, selectivity in tumor tissues and less toxicity compared to current platinum complexes. The diversity of ligands is crucial for these complexes to reach multiple biological targets and thus, overcome the limitation of contemporary therapies. One approach for multi-target molecules is the combination of bioactive ligands with recognized antitumor properties, such as natural naphthoquinones. Based on this, this study had proposed the evaluation and characterization of toxicogenetics and antitumor properties of eight new ruthenium complexes containing the natural ligands lapachol and lawsone. In general, the phosphinic series showed satisfactory antiproliferative activity against different cell lines, especially prostate carcinoma-derived cells. The IC50 values of these complexes were considerably lower than the values found for cisplatin. Furthermore, it was observed up to 28-fold selective for the complexes than cisplatin. Also, clonogenic assay suggests that the replacement of bipy ligand for phen increases the compound activity, inducing loss of DU-145 cells clonogenicity. Phosphinic complexes also described antiproliferative activities in three-dimensional (3D) cell models. In these assays, the lapachol-complexes have demonstrated cytotoxic activity independent of the hypoxia found in the spheroids. Besides, the results suggest that the DP-LP complex goes through the spheroid and destabilizes the cell aggregates, resulting in disruption of matrix-cell interactions. The DP-LP complex triggers apoptosis in DU-145 cells accompanied by an increase of cells in the G2 / M phase, which is consistent with the inhibition of AURORA-B protein activity. The central mechanism related to these effects is the presence of lapachol in DP-LP, which promotes the generation of intracellular reactive species that can cause DNA damage. DNA fragmentation, evidenced by comet assay, could activate the DNA repair systems that maintain the cells in the G2 / M checkpoint, due to a decrease of the AURKB gene expression. Also, DNA damage could activate caspases that conducted the DU-145 cells to apoptosis. In summary, the results found in this study contributed to the selection and elucidation of antiproliferative and cytotoxic properties of new compounds. We demonstrate the coordination of lapachol in a phosphinic ruthenium complex allowed the formation of compounds with potential impact in health targeting cancer treatment.
Recently, ruthenium complexes have occupied a leading position in the search for new anti-cancer therapies due to their potential of action, selectivity in tumor tissues and less toxicity compared to current platinum complexes. The diversity of ligands is crucial for these complexes to reach multiple biological targets and thus, overcome the limitation of contemporary therapies. One approach for multi-target molecules is the combination of bioactive ligands with recognized antitumor properties, such as natural naphthoquinones. Based on this, this study had proposed the evaluation and characterization of toxicogenetics and antitumor properties of eight new ruthenium complexes containing the natural ligands lapachol and lawsone. In general, the phosphinic series showed satisfactory antiproliferative activity against different cell lines, especially prostate carcinoma-derived cells. The IC50 values of these complexes were considerably lower than the values found for cisplatin. Furthermore, it was observed up to 28-fold selective for the complexes than cisplatin. Also, clonogenic assay suggests that the replacement of bipy ligand for phen increases the compound activity, inducing loss of DU-145 cells clonogenicity. Phosphinic complexes also described antiproliferative activities in three-dimensional (3D) cell models. In these assays, the lapachol-complexes have demonstrated cytotoxic activity independent of the hypoxia found in the spheroids. Besides, the results suggest that the DP-LP complex goes through the spheroid and destabilizes the cell aggregates, resulting in disruption of matrix-cell interactions. The DP-LP complex triggers apoptosis in DU-145 cells accompanied by an increase of cells in the G2 / M phase, which is consistent with the inhibition of AURORA-B protein activity. The central mechanism related to these effects is the presence of lapachol in DP-LP, which promotes the generation of intracellular reactive species that can cause DNA damage. DNA fragmentation, evidenced by comet assay, could activate the DNA repair systems that maintain the cells in the G2 / M checkpoint, due to a decrease of the AURKB gene expression. Also, DNA damage could activate caspases that conducted the DU-145 cells to apoptosis. In summary, the results found in this study contributed to the selection and elucidation of antiproliferative and cytotoxic properties of new compounds. We demonstrate the coordination of lapachol in a phosphinic ruthenium complex allowed the formation of compounds with potential impact in health targeting cancer treatment.
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Palavras-chave
Câncer, Lapachol, Complexo de rutênio, Apoptose, ERO