Estudo in silico e in vitro do redirecionamento farmacológico de um benzimidazol para o tratamento de Leucemia Mielóide Crônica quimiorresistente

Abstract

Chronic Myeloid Leukemia (CML) is a neoplasm among the most common for adult patients and the chemotherapy of this disease faces the problem of chemoresistance. Redirection and drug combination are powerful tools in this context and bring attention to families of compounds such as benzimidazoles, known for their antiparasitic effect, but which has already been studied for some neoplasms. Thus, the present work aims to attest to the cytotoxic potential of a benzimidazole derivative isolated and in combination with drugs used in the treatment of sensitive and resistant CML. It is worth mentioning that the name of the compound under study will be kept confidential until obtaining its legal protection, remaining on the acronym of BNZ for that purpose. Thus, the cell viability assay by Alamar blue of the treatments was carried out, in addition to the assessment of cell membrane integrity and mitochondrial membrane potential, by double marking IP / Rho123, and the analysis of cell cycle by cytometry. In addition, an in silico search for pharmacokinetic and pharmacodynamic characteristics was carried out, with the servers admetSAR, Molinspiration and SwissTarget Prediction, as well as molecular anchoring with Autodock 4.2.6. The isolated BNZ showed an IC50 of 106 nM for the sensitive strain (K562) and 1917 nM for the resistant strain (FEPS). The combination with Imatinib provided a 6-fold dose reduction index for the drug alone, demonstrating the effectiveness of this combination. Cytometry revealed that the isolated BNZ and its combination with Imatinib reduced the percentage of cells in the G0 / G1 phases, suggesting a washer for the Sub-G1 phase. In addition, the double label assay showed a reduction in the number of viable cells and an increase in the population of cells in late apoptosis / necrosis for treatments with BNZ. In silico assays revealed the ABL1 protein as a potential target for BNZ. Molecular docking against 3 ABL1 structures deposited in the protein structure database (PDBID: 2GQG, 2HYY and 5MO4), using Autodock 4.2.6, presented better results for the allosteric site (2GQG: -7.04 kcal / mol; 2HYY: -8.9 kcal / mol; 5MO4: -6.98 kcal / mol). This methodology also highlighted residues Ala433, Glu462 and Gly463 as the main ones for the interaction with BNZ at the allosteric site. Therefore, to date, the findings of this study suggest the anticancer potential of BNZ in CML through its interaction with ABL1 protein, as well as its synergistic effect in combination with Imatinib and Dasatinib.