Ten novel chalcone derivatives bearing thieno[3,2-d]pyrimidine moiety were successfully designed and synthesized based on the principle of drug molecular fragment and splicing principle and evaluate their antiproliferative activities. The structures of the target compounds were confirmed by 1H NMR, 13C NMR and MS. A549, HepG2 and PC-3 cell lines were employed to evaluate anticancer activities of the target compounds using MTT assay in vitro．All the target compounds showed excellent antiproliferative activities against all tested cancer cell lines. Among them, compound Ⅶh exhibited remarkable inhibitory activity against A549, HepG2 and PC-3 cell lines with IC50 value of 0.87 μmol/L, 2.43 μmol/L and 2.02 μmol/L, respectively, which were more potent than that of the positive control sorafenib. Chalcone derivatives bearing thieno[3,2-d]pyrimidine moiety showed excellent antiproliferative activities, and could serve as a scaffold for anticancer drug development.