Comprehensive assessment of 3-benzyloxyflavones as β-glucosidase inhibitors: <i>in vitro</i>, <i>in vivo</i>, kinetic, SAR and computational studies

In this study, a series of 3-benzyloxyflavone derivatives (1-10) was designed and, for the first time, evaluated for both <i>in vitro</i> and <i>in vivo</i> inhibitory activity against the β-glucosidase enzyme. The enzyme inhibitory potential of these derivatives was further assessed in an antihyperglycemic context using <i>in vivo</i> mechanism-based assays on <i>p</i>-nitrophenyl-β-d-glucopyranoside (PGLT) induced diabetic models. Additionally, structure-activity relationship (SAR) was employed to identify structural features crucial for activity. Molecular docking analyses revealed that both the potent compounds and co-crystallized ligands shared similar binding orientations within the active sites of β-glucosidase (PDB IDs: 3AJ7; 66K1). Molecular dynamics (MD) simulations validated the stability of the inhibitor-enzyme complexes under physiological conditions, while density functional theory (DFT) calculations helped elucidate electronic properties critical for activity. Drug-likeness analysis was also conducted to assess the pharmacokinetic potential of the derivatives. The results highlighted several derivatives with significant inhibitory activity, desirable pharmacokinetic profiles, and promising drug-like properties, making them potential candidates for therapeutic development. The target derivatives (1-10) demonstrated strong potential as lead compounds for developing new anti-diabetic agents with effective anti-hyperglycemic properties.