Malignant tumor is one of the major diseases that seriously threaten human health today. Compared with traditional chemotherapy, targeted drug therapy has become a new idea of tumor therapy. And EGFR (epidermal growth factor receptor) is highly expressed in many human tumor cell lines, which is a biomarker of tumor proliferation. In this paper, small molecule tyrosine kinase inhibitors with quinazoline structure aiming at EGFR were studied. A series of novel quinazoline derivatives (4a~4l) have been designed and synthesized from 4-hydroxyquinazoline as the parent core. Structures of target compounds were characterized by ¹H NMR and ¹³C NMR spectra. The in vitro anticancer activity of compounds 4a~4l was evaluated by MTT assay against Hela, MCF-7 and A549 tumor cell lines, and apoptosis-inducing capacity was investigated by Annexin-V/PI staining assay. The results showed that all compounds had good antitumor activity against the test tumor cell lines. Especially, compound 4a exhibited the best anticancer activity (IC₅₀ = 10.23 μM) against Hela cell lines, remarkable ability to induce apoptosis, and low toxicity, which identified 4a as a promising anticancer drug aiming at EFGR.