A new series of p-tertbutylcalix[4]arenes integrated with isomer trifluoromethyl anilines has been designed as potential anticancer agents. The integration of trifluoromethyl anilines to p-tertbutylcalix[4]arene allows the access of C4, C5, C6, C10, C11 and C12 analogues in good yields (more than 85%). To evaluate the antiproliferative activity of the prepared derivatives, the cancer cell growth inhibition assays have been performed for three cancer cell lines (HeLa, A549, MCF-7, Hep-2) and kidney normal epithelial cells (Vero). The compounds including CF3 groups at ortho position (C4 and C10) show 25 and 12 fold higher antiproliferative activity against MCF-7 cells over Vero cells, respectively. Also, C6 and C12 show 11 and 8 fold higher antiproliferative activity against HeLa cells over Vero, respectively. These results highlight that new calixarene derivates can be worthy of further studies as potential anticancer agents.