Heterocycles are an important class of compounds, making up more than half of all known organic compounds. Benzimidazole is a dicyclic compound having imidazole ring (containing two nitrogen atoms at nonadjacent positions) fused to benzene. Benzimidazole and its derivatives are used in organic synthesis. The main aim of this proposed research work is to design and synthesize a novel class of (E)-2-(2-(5-nitro-1H-benzo[d]imidazol-2-yl)vinyl)-4H-chromen-4-one derivatives and characterization by using IR, 1H NMR, 13C NMR, and mass spectroscopy, in vitro anticancer activity. Synthesized compound were evaluated for their in vitro anti cancer activity by SRB assay method against Human Cancer Cell line A-549. From the result the compound R1 and R6 allowed only 21.9 and 39.4 of % cell growth at the concentration of 80µg/mL. Whereas other compounds has showed 57-94% 0f %cell growth. From the results, it may conclude that the substitution on the Coumarin nucleus alter the biological activity. The un-substituted (R4) and Flouro (R6) and Chloro (R1) substituted Coumarin derivatives were found to be potent by producing biological activity.