The genus Anchusa L. (Boraginaceae) is represented by 15 species in the flora of Turkey. Anchusa species are used traditionally as wound healing and diuretic agent. In addition, some Anchusa species are used as demulcent, expectorant, analgesic, sedative, diaphoretic and antihypertensive purposes. Carminative, antidiabetic, hemostatic and antirheumatical usage is also recorded. The aim of this study is to determine in vitro antioxidant, anti-AChE, anti-BuChE and α-glucosidase activity of methanol extract (Methanol: water = 80:20, v/v) prepared from roots and aerial parts of Anchusa undulata L. subsp. hybrida (Ten.) Coutinho. Antioxidant activity was evaluated using ABTS and DPPH radical scavenging assay, total phenolic and total flavonoid content. IC50 values for DPPH were 239.47 and 292.04 µg/mL and for ABTS were calculated as 41.15 and 32.3 µg/ml. The total phenolic content of root and herbal extract were found as 130.39 and 122.94 mg/g, while flavonoid content as 65.25 and 117.5 mg/g. In the enzyme inhibition studies, the methanol extract showed cholinesterase and α-glucosidase enzyme inhibition activities with the increasing concentrations. The significant enzyme inhibition activities are depended on higher amount of phenolic compounds.