GC-MS-guided phytochemical profiling of Morus alba leaves identifies 5-O-methylnaringenin as a promising antidiabetic agent.

Objective: Type 2 diabetes mellitus (T2DM) is a rapidly increasing metabolic disorder linked with cardiovascular, renal, and neurological complications. This study investigated the phytochemical composition, antioxidant, antimicrobial, and antidiabetic potential of methanolic leaf extract of Morus alba using in vitro and in silico approaches. Methods: Gas chromatography-mass spectrometry analysis of the methanolic extract identified 16 bioactive compounds, including 5-O-methylnaringenin 2TMS derivative and Benzenepropanoic acid derivatives. Antidiabetic activity was evaluated through α-amylase and α-glucosidase inhibition assays, while antioxidant activity was assessed using the DPPH assay. Antimicrobial activity was tested against Bacillus subtilis and Staphylococcus aureus. Molecular docking, molecular dynamics (MD) simulation, SwissADME, and ProTox-II analyses were also performed. All in vitro experiments were conducted in triplicate (n = 3), and data were analyzed using one-way analysis of variance followed by Tukey's multiple-comparison test, with p < .05 considered statistically significant. Results: The extract showed concentration-dependent inhibition of α-amylase (IC50 = 184.6 µg/mL) and α-glucosidase (IC50 = 112.3 µg/mL), with stronger inhibition of α-glucosidase. DPPH analysis confirmed antioxidant activity (IC50 = 230 µg/mL). Antimicrobial assays demonstrated inhibitory effects against both tested bacterial strains. Molecular docking identified 5-O-methylnaringenin as the most active compound with stable binding interactions at the α-glucosidase active site, supported by 100 ns MD simulation. SwissADME predicted favorable gastrointestinal absorption, blood-brain barrier permeability, and compliance with Lipinski's rules, while ProTox-II suggested no organ-specific toxicity. Conclusion: The findings indicate that M. alba leaves possess promising antidiabetic, antioxidant, and antimicrobial properties. Among the identified compounds, 5-O-methylnaringenin demonstrated strong therapeutic potential with favorable pharmacokinetic and safety characteristics, supporting further in vivo studies for T2DM management.