The Influence of Para-Substituent on the Molecular Properties and Biological Activities of Aroylhydrazone Derivatives: DFT Analysis and Anti-Cervical Cancer Evaluation Through In Silico and In Vitro Studies

The synthesis of four aroylhydrazone-derived compounds was successfully conducted via the condensation reaction between benzoyl hydrazine and four different aromatic aldehydes. The resulting comppunds were identified as 4-Methoxybenzylidene ben-zo hydrazide (L1), 4-hidroksi-benzaldehid benzoyl hydrazone (L2), 4-methylbenzylidene benzohydrazide (L3) and 4-nitrobenziliden-2-benzoyl hydrazine (L4). Their spectroscopi cal and physicochemical characteristics were analyzed through UV-Vis, IR,¹H NMR, and¹³C NMR instruments. Structural optimization was subsequently performed using density functional theory (DFT), followed by theoretical evaluations of UV-Vis and IR spectral modeling, reactivity and stability, and several quantum chemical descriptors. It was revealed that the theoretical results agreed with the experimental yields. The results of DFT analysis showed that L1 (ΔE_GAP = 4.58 eV) and L3 (ΔE_GAP = 4.42 eV) have high molecular stability and reactivity. These parameters indicate L1 and L3 as candidates for anticancer compounds. Besides, the antioxidant and cytotoxicity of the prepared compounds were observed to elucidate their specific mechanisms. L1 and L3 showed moderate anticancer activity with IC₅₀ values of 82.12 and 42.64 µg/mL, respectively. In addition, insilico analysis, indicated strong binding affinities for L1 and L4, with docking binding energies of −9.37 and −10.57 kcal/mol. The Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) predictions and pharmacokinetic parameters of the samples further supported their favorable oral bioavailability.