A series of novel 4-anilinoquinazoline derivatives with (E)-propen-1-yl moiety were designed, synthesized and evaluated for biological activities in vitro. Most compounds exhibited highly antiproliferative activities against all tested tumor cell lines including A431, A549, NCI-H1975 and SW480 cells. Especially, compound 6e not only presented strong antiproliferative activities against the tested four tumor cell lines (IC50 of 1.35, 8.83, 5.53 and 6.08 μM, respectively) which expressed wild type or L858R/T790M double mutant epidermal growth factor receptor (EGFR), but also showed potent inhibitory activity against wild type EGFR (IC50 = 20.72 nM). The result of molecular docking with EGFR suggested the binding mode of 6e was similar to gefitinib, but different from lapatinib. Additionally, western blot analysis showed that 6e inhibited the phosphorylation of EGFR and its downstream signaling proteins in lung cancer cells. The work could be very useful starting point for developing a new series of tyrosine kinase inhibitors targeting EGFR.