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We have isolated anti-diabetic activity of bioassay guided active fraction from Anisomeles malabrica, which was increased the glycemic control and insulin levels, regulated the carbohydrate and lipid metabolism and protected from the oxidative stress. An interesting effect of the active fraction has enhanced the expression of Hepatic Glucose Transporter (Glut-2) and down-regulated the Hepatic Phosphoenolpyruvate Kinase (PEPCK) enzyme. Further using UHPLC-MS analysis of active fraction, we identified that the phytoconstituents Luteolin, Caffeic acid, Kaempferol, and Chlorogenic acid are responsible for glycemic controlling and/or enhance the secretion of insulin from beta cells of the pancreas.