Butt_2026_Naunyn.Schmiedebergs.Arch.Pharmacol__

Cholic acid-derived hydrazones represent a promising scaffold for multifunctional drug discovery. We synthesized and characterized two novel cholic acid-based hydrazone derivatives: DFBCH (2,4-difluorobenzylidene conjugate) and MEBCH (3-methoxybenzylidene conjugate) through various spectroscopic techniques, confirming the structural integrity of the synthesized compounds, followed by their comprehensive biological evaluation. In vitro screening showed a greater antibacterial zone of inhibition of MEBCH compound as compared to DFBCH against the tested strains. MEBCH demonstrated potent antioxidant activity (IC(50) 1.87 +/- 0.20 mg/mL) relative to ascorbic acid (IC(50) 7.3 +/- 1.40 mg/mL) by DPPH assay illustrating superior antioxidant activity. Both the compounds exhibited greater % inhibition against Hela cell line as compared to doxorubicin, suggesting greater inhibitory activity. MEBCH was the superior inhibitor of acetylcholinesterase (IC(50) 1.67 +/- 0.61 microM), surpassing the standard donepezil (IC(50) 3.13 +/- 0.1 microM), and demonstrated effective tyrosinase inhibition (IC(50) 1.87 +/- 0.88 microM), comparable to kojic acid (IC(50) 2.38 +/- 0.75 microM). Both of these compounds demonstrated effective beta-glucosidase inhibition with an IC(50) = 2.23 microM, comparable to standard miglustat (IC(50) 2.02 +/- 1.05 microM). Molecular docking (Glide) revealed MEBCH producing a favorable docking score with AChE (- 6.711; E-model - 57.480), indicating a stable predicted binding pose relative to DFBCH. BioTransformer 3.0 predicted CYP 450-mediated pathways for both analogues; docking of predicted metabolites showed retained or improved affinity (MEBCH metabolite to AChE - 6.3 kcal/mol) and lower RMSD bounds (AChE; lb/ub = 5.8/6.2 A; beta-glucosidase lb/ub = 5.25/5.73 A). DFT calculations revealed small HOMO-LUMO gaps and solvent-sensitive dipole moments consistent with moderate chemical reactivity and solvent stabilization. Single-dose oral pharmacokinetics (10 mg/kg) in rats revealed MEBCH with higher C(max), AUC, longer t(1/2), and greater oral bioavailability than DFBCH. Collectively, these data nominate MEBCH as a lead cholic-hydrazone for further preclinical development against enzyme targets relevant to neurological and oxidative-stress pathways.