In this study, we synthesized a series of novel N-acetyl Schiff bases (6a–e) 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 6a, 6d, and 6e, with IC₅₀ values of 73.25, 49.80, and 111.73 μM, respectively, in DU145 cells. Notably, 6d exhibited a 10-fold selectivity toward cancer cells over normal cells. Enzyme inhibition studies demonstrated that compound 6d exhibited the most potent inhibitory activity against the carbonic anhydrase isoforms hCAI and hCAII, with the lowest recorded IC₅₀ and K _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 6d 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 6d 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 6d, 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 6d holds significant promise as a potent dual-target inhibitor with applications in prostate cancer therapy and glaucoma treatment.

In this study, the structure of 4-[4-(diethylamino)-benzylideneamino]-5-benzyl-2 H -1,2,4-triazol-3(4 H )-one (DBT) was examined through spectroscopic and theoretical analyses. In this respect, the geometrical, vibrational frequency, ¹ H and ¹³ C-nuclear magnetic resonance (NMR) chemical shifts, thermodynamic, hyperpolarizability, and electronic properties including the highest occupied molecular orbital–lowest unoccupied molecular orbital (HOMO–LUMO) energies of DBT as a potential non-linear optical (NLO) material were investigated using density functional theory at the B3LYP level with the 6-311G basis set. ¹ H and ¹³ C-NMR chemical shifts of DBT with the gauge-invariant atomic orbital and continuous set of gauge transformation methods (in the solvents) were estimated, and the computed chemical shift values displayed excellent alignment with observed ones. Time-dependent density-functional theory (TD-DFT) calculations with the integral equation formalism polarizable continuum model within various solvents and gas phases in the ground state were used to evaluate UV-vis absorption and fluorescence emission wavelengths. Thermodynamic parameters including enthalpy, heat capacity, and entropy for DBT were also calculated at various temperatures. Moreover, calculations of the NLO were carried out to obtain the title compound’s electric dipole moment and polarizability properties. To illustrate the effect of the theoretical method on the spectroscopic and structural properties of DBT, experimental data of structural and spectroscopic parameters were used. The correlational analysis results were observed to indicate a strong relationship between the experimental and theoretical results.