Eleven novel 4-anilinoquinazoline derivatives were synthesized and evaluated for their in vitro antiproliferative activity. Among them, compound 9a exhibited the best potency, with IC values of 25-682 nM against various types of cancer cell lines. In addition, 9a was confirmed that it could arrest the cell cycle at G /M phase and trigger apoptosis. Indirect immunofluorescence staining revealed its anti-tubulin property. Importantly, 9a significantly inhibited tumor growths in SM-7721 xenograft models (57.0% tumor mass change) without causing significant loss of body weight, suggesting that 9a is a promising new anticancer agent to be developed. This article is protected by copyright. All rights reserved.