Abstract
Oral antidiabetic drugs mainly act by stimulating pancreatic β-cells, but prolonged stimulation may lead to exhaustion and impaired insulin secretion. In contrast, AMP-activated protein kinase (AMPK) lowers blood glucose through an insulin-independent pathway, offering therapeutic potential. This study aims to identify AMPK activators in oyster mushrooms and fenugreek using GC-MS and bioinformatics analysis. The bioactive compounds in both samples were identified through GC-MS analysis. Molecular docking, pharmacokinetics, drug-likeness, toxicity, and molecular dynamics simulation (MDS) were employed to evaluate the therapeutic potential of the identified phytochemicals. The results showed that the ester class of compounds was the most abundant in both fenugreek and oyster mushroom extracts, identified by GC-MS analysis. Molecular docking revealed that the selected ligands exhibited strong binding affinities to the AMPK target protein. Pharmacokinetic and drug-likeness analyses confirmed that most compounds met acceptable ranges for potential AMPK activators. Molecular dynamics simulations further demonstrated that the selected ligands formed more stable ligand-protein complexes with fewer fluctuations compared to the control drug, glibenclamide. Overall, this study suggests that 2',6'-Dihydroxyacetophenone, 1-Methyl-2-phenylbenzimidazole, and N-Methylphthalimide are potential AMPK direct activators capable of mitigating Type 2 diabetes through an insulin-independent pathway. However, further in vitro and in vivo validation is required.