Abstract: The purpose for this study was to evaluate the biological, antioxidant, anti-inflammatory and anti-cancer properties of Calotropis procera alcoholic extract and its various solvent subfractions. The total antioxidant property was estimated by the Ferric Reducing Antioxidant Power (FRAP), 1,1-diphenyl-2-picrylhydrazyl (DPPH·), 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS·+) and β-carotene bleaching tests. Among the ethanol extract and three fractions, the ethyl acetate fraction showed the highest phenolic content (108.26 mg gallic acid g-1) and the best antioxidant activity. The ascorbic acid equivalent antioxidant capacities of the ethyl acetate fraction were 0.26, 0.53, 0.17 mmol g-1 in FRAP, ABTS and DPPH assays, respectively. This study verified that the ethanol extract and ethyl acetate fraction from Calotropis procera have strong antioxidant activity that were correlated their high level of phenolic content. Furthermore, the same extract showed appreciable anti-inflammatory via lipoxygenase inhibitory activity (LOX), IC50 values ranged from 7.12-52.68 μg mL-1. Moreover, ethyl acetate fraction showed the strongest cytotoxic effect (IC50 = 10.6 μg mL-1) against MCF-7 breast cancer cell line.