Three new metabolites, 5-hydroxy-3,7-dimethoxyflavone-40 -O-b-glucopyranoside (1), 2b,19-epoxy-3b,14b-dihydroxy-19-methoxy-5a-card-20(22)-enolide (4) and b-anhydroepidigitoxigenin-3b-O-glucopyranoside (5), along with two known compounds, uzarigenine (2) and b-anhydroepidigitoxigenin (3), were isolated from Calotropis procera (Asclepiadaceae). The structure elucidation was accomplished mainly by nuclear magnetic resonance (NMR) spectroscopic and mass spectrometric methods. To examine putative antimicrobial or cytotoxic activities, various bioassays were performed. Uzarigenine (2) demonstrated moderate cytotoxicity.