In search of new class of cytotoxic molecules we have designed and synthesized novel scaffold by linking nitric oxide donating group to acridone moiety. Synthesized derivatives were evaluated for their cytotoxic potentials in comparison with reference drugs doxorubicin (DX) against Human colon cancer cell line (HT-29) by using Sulphorhodamine-B (SRB) assay. Compound 15 exhibited good cytotoxic activity with an IC50 value 8.6 μM. Cytotoxic effects are presumed to be due to the presence of nitric oxide donating group, acetyl group on 2nd position and butyl side chain at N10 position. Further, we tested whether the levels of NO production by the indicated compounds in human colon cancer cell line were associated with their cytotoxic effects. Good correlations were observed when cytotoxic data of these molecules compared with nitrite production.