Prostate cancer is the third most diagnosed cancer worldwide, and the second cause of cancer deaths in men. The currently available treatments are not always effective and may be associated with unwanted side effects. The process of developing new drugs is expensive and can take several years. Thus, drug repurposing emerges as an interesting alternative since it uses clinically studied and available drugs for a new clinical use. The present study aimed to explore the effects of a β-blocker (carvedilol), a selective serotonin reuptake inhibitor (sertraline) and an antimetabolite drug (5-fluorouracil), alone or in binary mixtures, on the cancer cell line (22Rv1) as well as on the normal prostate cell line (PNT-2) cell viability. Overall, the tested conditions demonstrated the ability of the drugs to induce toxic effects and allowed the estimation of median lethal concentrations. The cell line 22Rv1, compared to the normal cell line, was more sensitive to sertraline and 5-fluorouracil but more resistant to carvedilol. Data from combined exposures conditions demonstrated the potential value of these substances