Sayar_2026_Chem.Biol.Drug.Des_107_e70285

A series of benzimidazoles and its analogs (benzoxazoles and benzothiazoles) bearing a benzohydrazide-hydrazones moiety at the 2-position were designed and synthesized as potential acetylcholinesterase (AChE) inhibitors. Cholinesterase (ChE) inhibitory activity results showed that all compounds displayed good inhibition of AChE, whereas only some of the compounds were active against BChE. Among the title compounds, it was found that all benzimidazole series were selective towards AChE. Kinetic analysis and molecular modeling studies were conducted on the most active compounds, namely 1j for AChE and 3g for BChE inhibition. The kinetic results showed that the tested compounds exhibited a mixed-type inhibition mechanism on both enzymes. In cell culture studies, all compounds were evaluated for their neuroprotective effects against hydrogen peroxide (H(2)O(2))-induced oxidative stress in the SH-SY5Y human neuroblastoma cell line. Several compounds demonstrated more than 30% neuroprotection at a low concentration (1 microM) in the presence of H(2)O(2). Selected compounds, chosen based on their EC(50) values, were further examined for their effects on glycogen synthase kinase-3 beta (GSK3beta), an enzyme whose activity has been reported to be increased in the brains of patients with Alzheimer's disease (AD). The inhibition of GSK3beta was assessed by measuring phosphorylated GSK3beta (pGSK3beta, Ser9) protein expression via Western Blot analysis under oxidative stress conditions. Among the tested compounds, 1l, 2b, 2j, and 3l significantly increased pGSK3beta (Ser9) protein levels compared to cells treated with tideglusib, a known GSK3beta inhibitor.