Prostate cancer is a prevalent malignancy that ranks second in both incidence and mortality rates among males. Although different treatment options are available, they have drawbacks (e.g., limited specificity, high cytotoxicity, and development of resistance). To address these shortcomings, researchers are focusing on more efficient alternative strategies. One of such approach involves the development of more efficient drug delivery systems. Polymeric nanoparticles (NPs), particularly those composed of bovine serum albumin (BSA), are emerging as a promising, environmental-friendly strategy for targeted drug delivery. These NPs offer several advantages, including biocompatibility, stability, and biodegradability. The main objective of this study was to synthesize BSA nanoparticles, using a genipin-based crosslinking method, to be used as carriers for two compounds: an antimetabolite (5-fluorouracil) and an antidepressant (sertraline), which have shown potential efficacy in the treatment of prostate cancer. The effects of the synthesized particles (with and without loaded drugs) on the PNT-2 cell line (normal prostate cells) and the 22Rv1 cell line (prostate cancer cells) viability to validate its potential utility. Of the synthesized particles, BSA NPs with a size range of 200–400 nm with low size variability (polydispersity index < 0.2) were selected for encapsulation procedures. BSA NPs and genipin exhibited non-toxicity in both cell lines, while sertraline-loaded BSA NPs showed a nullifying effect. These results highlight the promising potential of BSA NPs for biological applications.