In silico studies and cell-based assays to identify new therapeutic approaches to prostate cancer

Abstract

The prostate cancer has the highest incidence among all neoplasia in middle-age men. This pathology is hormone-dependent, being androgen receptor (AR) the target of choice for antitumoral drugs bicalutamide, flutamide e nilutamide. However, the androgen-ablation therapy may not be efficacious at the late-phase disease, and it is necessary to introduce AR-independent bioactive compounds. In this case, some compounds that target the apoptotic process, being under clinical studies. This work aims to identify AR-dependent or AR-independent new chemical entities (NCEs) that inhibit the AR-signaling pathway, or by leading to cell death independently of AR (through the modulation of mTOR e survivin cell signaling) in order to identify new therapeutic approaches to prostate cancer. The rational drug discovery workflow is based on the ligand (LBVS) and structure (SBVS) virtual screenings using AR, surviving and mTOR, in conjunction with the prediction of pharmacokinetics properties. It is estimated that 30 to 40 molecules will be cherry-picked with the aid of the computational methods to be studied by cell-based assays. These compounds will be tested by flow cytometry to observe the phenotypic alterations of two cell models of prostate cancer (LNCaP and DU 145) regarding the intracellular granularity, in a HCS-type study, together with the analysis of the cell cycle arrest. All phenotypic alterations will lead to activity "fingerprints" observed to a selected reference set (composed by a mix of drugs and bioactive molecules). Structure-activity relationships (SAR) will be devised to optimize the compound series and to analyze the models and hypotheses established during the in silico step. The cell-based assays will be cornerstone to the application of polypharmacology and biochemical networks concepts at the forefront of the drug discovery process, with the aim of integrating the medicinal chemistry knowledge to exploit the chemical-biological space of prostate cancer. (AU)