A new series of thirty-six dihydroxylated 2, 6-diphenyl-4-aryl pyridines containing hydroxyl groups at the orho, meta, or para position of 2- and 6-phenyl rings attached to the central pyridine were designed and synthesized. They were evaluated for topoisomerase I and II inhibitory activity and cytototxicity against several human cancer cell lines for the meta or para position of 2- or 6-phenyl ring in combination with thienyl or furyl group at 4-posintion of central pyridine displayed significant topoisomerase II inhibitory activity and cytotoxicity. Positive correlation between topoisomerase II inhibitory activity and cytotoxicity was observed for the compounds 9-11, 15-17, 19, 21-23, 28, and 41. Among all the synthesized compounds, compound 17emerged as the most promising topoisomerase II inhibitor with significant cytotoxicity. ⓒ2015 Elsevier Ltd. All rights reserved.