Background: The traditional information of Acalypha alnifolia is captivate towards its therapeutic efficacy against diabetes along with the scientific confirmation were leads to an instinct to extract phytocompounds and to evaluate antidiabetic-molecular mechanism. Methods: Column chromatographic compound isolation were adopted to extract the pure compounds from bioactive leaf extract and interactions of the compounds with diabetic metabolic proteins were accomplished through in-silico docking studies. By the direction, the anti-diabetic property was valued by the expressions of Glycogen Synthase Kinase 3β(GSK 3β) and Insulin on insulin-Receptor on Hep G2 cells. Results: The flavone compound - 5,7-dihydroxy-2-(4-hydroxyphenyl)-3-methoxy-4H-chromen-4-one and pyridine alkaloid - (2,6-dihydroxypyridin-4-yl)(3,5-dihydroxytetrahydro-2H-pyran-4-yl) acetic acid were isolated. Both the compounds reduced the GSK 3β expression and the flavone contributed also to enhance insulin binding ability expediently. Conclusions: The results concluded that the compounds from A. alnifolia having potent to maintain glucose homeostasis in liver insulin-metabolism. By exploring pharmaceutical importance of the isolated compounds, this study will be an important breakthrough in developing new drug for Diabetes. © 2022 The Author(s)