Objective：To explore anti-cancer activities of a novel bis-aryl urea compound（code name：N69B）using non-small cell lung cancer A549 cells. Methods：The non-small cell lung cancer A549 cells were choosed and treated by N69B at different concentrations. The cell proliferation was quantified by CCK-8 method；the cell cycle was determined by flow cytometry；the apoptosis rate were detected by Annexin V/PI double staining；and the relative expression of Cleaved caspase-3，8，9 was measured by the Western blot. Results：Compared with the DMSO control group，N69B inhibited the proliferation of A549 cells in manners that are time-and dose-dependent（P < 0.05 or P < 0.01）. Twenty-four hours after the treatment with N69B at 0.5，1.0 and 1.5 μM，respectively，the number of cells in G2 phase significantly increased（P < 0.01）；the apoptosis rate increased in different degrees；Cleaved caspase-3，8，9 expression showed a dose-dependent elevation. Conclusion：N69B inhibits the proliferation of A549 cells，possibly through cell cycle arrest in G2-M phase and apoptosis induced by caspase activation.