<p>Diabetes is a 21st-century epidemic affecting 10.1 crores Indians as of 2023. Ayurveda, a traditional indigenous medical practice, can be leveraged to manage diabetes, as the allopathic antihyperglycemic drugs are reported to cause severe repercussions. This study employs a combinatorial integrated approach to explore the scientific anti-diabetic prophylaxis of Madhumehantak Churna, an Ayurvedic formulation. Hydroalcoholic extract of Madhumehantak Churna (MCH) exhibited robust in-vitro enzyme inhibition and antioxidant assays. Out of 36 major phytocompounds identified by LC–MS profiling in MCH, ProTox 3.0 identified 29 phytocompounds with no significant toxicity, screened based on their toxicity (LD₅₀). These were subjected to integrated Network pharmacology and bioinformatics, involving in-silico molecular docking, ADMET, homology modelling and simulation studies. In-silico molecular docking studies of 29 compounds with 28 proteins, including GLUT12, Myeloperoxidase, Phosphodiesterase 4D, Cathepsin B, Cathepsin S identified from literature studies and network pharmacology, devised by STRING and CYTOSCAPE, highlighted that Ptelatoside A showed the strongest binding affinities. In-silico pharmacokinetics and ADMET analysis predicted Ptelatoside A, a glucoside, to possess the requisite drug likeness parameters. GROMACS-assisted Molecular dynamics simulation (MDS), PCA and KEGG pathway identified insulin resistance and insulin signalling pathways as the probable underlying molecular actions of Ptelatoside A against diabetes mellitus. The combinatorial study involving in-vitro anti-diabetic assessments, network pharmacology, molecular docking, in-silico pharmacokinetics, PCA analysis and simulation studies, underscores that the glycoside Ptelatoside A from the Ayurvedic formulation, Madhumehantak Churna, can be a future solution for anti-hyperglycemic drug development.</p> Graphical abstract <p></p>