Planejamento, síntese e avaliação anticâncer de derivados de lapachol e umbeliferona

Abstract

Cancer is the second cause of death in the world and is considered a public health problem. Cancer is the second leading cause of death in the world and is considered a public health problem. Due to this scenario, the search for substances that possess anticancer potential from compounds of natural or semi-synthetic origin is increasing, as well as the use of in silico methods in order to predict pharmacokinetic, toxicological and biological activity of molecules for rational planning of drugs. The objective of this research was to plan, synthesize and evaluate the anticancer potential of lapachol and umbelliferone derivatives, associating in silico and in vitro methods in order to potentiate the chance of finding bioactive compounds. 25 umbelliferone derivatives and 25 lapachol derivatives were designed, which were evaluated by molecular anchoring using the Autodock vina® software having topoisomerase I and II enzymes with biological targets, as well as their molecular, toxicological and pharmacokinetic properties evaluated by through Molinspiration®, Osiris® and PreADMET® software. The selected derivatives were synthesized and confirmed by NMR analysis and mass spectrometry and evaluated in vitro for their anticancer potential by cytotoxicity, genotoxicity analysis that was evaluated by the alkaline pH comet assay, which detects any damage caused to the DNA, of neutral pH, specific for breaks of the double tape and damage caused to the DNA by topoisomerase inhibitors I and II, as well as the evaluation of the gene expression of these enzymes. After the in silico analysis, the derivatives LP04, LP017, LP18, LP20, LP23, LP24, LP25, UMB22, UMB24 and UMB25 were selected because they presented a better complementarity profile with the active site of topoisomerase I and II, as well as interactions with critical amino acid residues characteristic of the inhibitors of these enzymes. In addition, they presented good molecular profiles, pharmacokinetics and low toxicological effect, characteristics desired in the planning of drugs. The cytotoxicity evaluation was performed by the Alamar Blue method, the derivatives LP04, LP17, LP18, LP20 and LP25 presented cytotoxic activity in the breast cancer (MCF-7) and colorectal (HCT116) strains, being selected for the subsequent tests, exhibiting an IC 50 in 72 hours for the MCF7 strain ranging from 2.41 to 8.20 μM and from 5.40 to 18.68 μM in the HCT116 strain, all derivatives having better activity when compared to the starting molecule . Both the umbelliferone and the starter molecules showed cytotoxic activity greater than 20 μM, and did not present an anticancer potential. Through the analysis of the damage index, it was observed that the derivatives presented significant damage (p <0.05) in alkaline pH, neutral pH and in the analysis of induction of DNA damage by topoisomerase inhibitors, presenting a higher index of neutral comet, which ranged from 37.6 ± 0.76 to 83.0 ± 3.5 in the HCT116 line and from 40.6 ± 2.7 to 105.3 ± 2.0 in the MCF7 strain suggesting that to act as inhibitors of topoisomerase II. In the analysis of topoisomerase gene expression, only the activity of LP25, which showed a reduction in the expression of the topoisomerase I, II alpha and II beta genes at the concentration of 7.5 μM and 15 μM, could be evaluated. Thus, the derivatives showed anticancer potential and more specific studies were necessary to evaluate topoisomerase inhibition directly as well as to evaluate other pathways of inhibition and mechanisms of death of these derivatives.