A novel series of five 7-piperazin-substituted [1,3]oxazolo[4,5-d]pyrimidines have been synthesized and characterized by Infrared (IR), Proton Nuclear Magnetic Resonance (1H-NMR), Carbon-13 Nuclear Magnetic Resonance (13C-NMR) spectroscopy, elemental analysis and chromato-mass-spectrometry. The anticancer activities of the all the newly synthesized compounds were valuated via single high dose (10-5M) against 60 cancer cell lines by the National Cancer Institute according to its own screening protocol. In the next phase, the compounds have been selected for five-dose assay. Among these compounds 5-phenyl-7-piperazin-1-yl-2-p-tolyl[1,3]oxazolo[4,5-d]pyrimidine displayed the most growth inhibitory (GI50 was in range of 0.2−2.0 μM), cytostatic (TGI − 0.3-4.2 μM) and cytotoxic (LC50 − 0.6-7.8 μM with the exception of Leukemia CCRF-CEM cell line, LC50>100 μM) activities against all cancer cell lines. The most selectivity 5-phenyl-7-piperazin-1-yl-2-p-tolyl[1,3]oxazolo[4,5-d]pyrimidine demonstrated against Leukemia and Colon Cancer subpanels. These results provided evidence that compound could be useful for developing new anticancer drugs.