Four compounds were isolated from the aerial parts of Justicia spicigera Schltdl. (Acanthaceae) and were identified as uracil (1), uridine (2),megastigmane vomifoliol-9-O-β-D-Glucopyranoside (3) and β-sitosterol (4). The structures of these compounds were characterized by spectroscopic methods (IR, ¹H-NMR, APT, 2D NMR and EI/MS). The different crude extracts showed selective inhibitory activity against Mycobacterium tuberculosis (TB), remarkable cytotoxic activities against Human breast cancer (MCF-7) cell lines and Human lung carcinoma (A-549) cell lines and weak antimicrobial activities. This is the first report concerning the isolation of compounds (1-3) from genus Justicia.