Policetídeos citotóxicos da esponja marinha Plakortis angulospiculatus

Abstract

As part of a bioprospecting study to identify compounds from marine organisms with anticancer potencial compounds from the brazilian coast, we investigated the cytotoxicity of an ethanolic extract derived from the marine sponge Plakortis angulospiculatus, widely known as a source of cyclic endoperoxides with various pharmacological activities, including cytotoxic. The bioguided fractionation of the extract led to the isolation of 4 polyketides with a dihydrofuran ring (3, 4, 5, 6), 5- (1) and 6- membered cyclic endoperoxides (2, 7-10), including three new plakortides, 7,8-dihydroplakortide E (1), 2 and 10, and known compounds such as spongosoritin A (5), plakortide P (9). To compare the cytotoxicity of both groups in human tumor cells of colon (HCT-116), prostate (PC-3M) and non-tumor (MRC-5) by the MTT assay, we observed different activity profiles, as compounds that contained a dihydrofuran ring were generally less activeand displayed a time-dependent activity. Six membered cyclic plakortides, on the other hand, exhibited greater cytotoxicity and time-independent effect. The modes underlying the cytotoxic actions of 2, 3, 5, 7, and 9 were further investigated using HCT-116 cells. Flow cytometry studies demonstrated cell density reduction accompanied by a reduction in membrane integrity at higher concentrations, with the exception of 2. Further inspection through cell cycle analyses indicated that 3 and 5 induced blockage at G0/G1. To the contrary, 2, 7 and 9 delivered a G2/M arrest and presence of mitotic figures, however this effect was independent of microtubule polymerization, as demonstrated by confocal microscopy. The evaluation of plakortides 2 and 9 on breast cancer cell line (MCF-7) revealed no changes on cell cycle, but increased DNA fragmentation, indicating cell death by apoptosis. Using the trypan blue assay, we verified a synergistic effect between plakortide P (9) and cyclosporin A, a calcineurin inhibitor, which induced a decrease of over 80% in viability of HCT-116 cells when compared to cells treated only with plakortide P (9) or cyclosporin A, which suggests the involvement of calcium-dependent pathways in the effect presented by plakortide P.