The current work describes the ultrasonication assisted extraction followed by comprehensive investigation of antioxidant activity and α-amylase inhibition potential of hydroethanolic leaf extracts of Vitexnegundo. Freeze drying assisted ultrasonicated extraction revealed highest level of total phenolic contents (258.29±2.52 mgGAE/g DE) and total flavonoid content (155.91±1.75 mgRE/g DE) in 60% hydro-ethanolic V. negundo leaf extract. Furthermore, total antioxidant power (254.35±1.56 mg AAE/g DE) and antidiabetic effectbased on α-amylase inhibitory assay (IC50=42.91±1.24 μg/mL) was also depicted to be highest in the same extract. Based on current findings, 60% ethanolic extract of V. negundo leaves was revealed to be most potent for its antioxidant and α-amylase inhibition activity, hence could be used for formulation of new plant based functional food sornutraceuticals to improve human health.