Strategic Nanosuspension of Darolutamide for Enhanced Dissolution, Bioavailability, and Potentiated Prostate Cancer Treatment: Insights from Multiscale Computational Modeling

Darolutamide (DA), a nonsteroidal antiandrogen, demonstrates significant potential as a targeted therapeutic agent for prostate cancer; however, its clinical application is limited due to its poor aqueous solubility and low oral bioavailability. To overcome these limitations, we developed a nanosuspension-based formulation (DA-NS) with a mean particle size (PS) of 143.36 ± 5.24 nm, which, upon lyophilization, yielded nanocrystals (DA-NC) with a mean PS of 156.26 ± 7.58 nm, aimed at enhancing the solubility and dissolution rate. Molecular dynamics simulations provided mechanistic insights into the stabilization and formation of DA-NS at the atomic level. In vitro studies demonstrated superior antitumor activity of DA-NC in PC3 prostate cancer cells, characterized by enhanced cellular internalization, nuclear condensation, and apoptotic morphology under fluorescence microscopy. Mechanistic investigations further validated its pro-apoptotic action via elevated reactive oxygen species generation and induction of DNA damage. Western blot analysis confirmed the molecular signature of apoptosis, showing downregulation of antiapoptotic Bcl-2 and upregulation of caspase-3. Pharmacokinetic evaluation revealed a 5.81-fold increase in <i>C</i>_max and a 3.33-fold enhancement in AUC_0-48 over pure DA, confirming a significant improvement in oral bioavailability (<i>p</i> < 0.0001). Importantly, DA-NC exhibited a favorable safety profile, underscoring its potential as a biocompatible and efficacious oral therapeutic strategy for advanced prostate cancer.