Synthesis and Computational Evaluation of <i>N</i>-Acetyl-Derived Schiff Bases Incorporating 1,2,4-Triazoles for Dual Inhibition of Prostate Cancer Cells and Carbonic Anhydrases

In this study, we synthesized a series of novel <i>N</i>-acetyl Schiff bases (<b>6a</b>-<b>e</b>) containing 1,2,4-triazole moiety and evaluated their potential as anticancer agents through both experimental and computational approaches. Cytotoxicity assays on prostate cancer (PC) (DU145) and normal epithelial cells (PNT1a) demonstrated selective inhibition, particularly for compounds <b>6a</b>, <b>6d</b>, and <b>6e</b>, with IC₅₀ values of 73.25, 49.80, and 111.73 μM, respectively, in DU145 cells. Notably, <b>6d</b> exhibited a 10-fold selectivity toward cancer cells over normal cells. Enzyme inhibition studies demonstrated that compound <b>6d</b> exhibited the most potent inhibitory activity against the carbonic anhydrase isoforms hCAI and hCAII, with the lowest recorded IC₅₀ and <i>K</i> _i values (7.12 and 9.26 μM for hCAI, and 10.62 and 11.72 μM for hCA II, respectively), suggesting strong potential for antiglaucoma therapeutic application. To elucidate molecular interactions, QM/MM molecular docking highlighted the strong affinity of compound <b>6d</b> for the active sites of CYP17A1, hCAI, and hCAII enzymes. The coordination of functional groups with key residues, particularly the Zn²⁺ ion and HEM group, was confirmed by detailed binding analyses. Molecular dynamics simulations further validated the stability of these interactions over a 100 ns trajectory, with <b>6d</b> maintaining robust engagement with the protein targets. This stability was reflected in consistent RMSD and RMSF profiles, with minimal fluctuations, particularly in CYP17A1 complexes, suggesting a stable binding conformation. The Markov State Model (MSM) analysis, integrated with TICA-FES and MM-GBSA calculations, revealed rapid conformational stabilization of <b>6d</b>, especially in CYP17A1 complexes. The observed deeper energy wells in diffusion maps indicate stronger binding affinities and reduced conformational transitions compared to reference inhibitors, such as abiraterone and acetazolamide. These computational insights align with experimental findings, suggesting that <b>6d</b> holds significant promise as a potent dual-target inhibitor with applications in prostate cancer therapy and glaucoma treatment.