Cancer is a major health problem in the developed and developing Countries.Inspired by the wide range of useful activities possessed by the acridine derivatives and in continuation of our efforts in search of potential anticancer agents, we have synthesized 9-anilinoacridine derivative and 9-anilino-2-methoxy acridine derivatives. The newly synthesized compounds evaluated for anticancer activities against two cancer cell lines i.e. human small cell lung cancer (A-549) cell line and human endothelial cervical cancer (HeLa) cell line in vitro by MTT assay. The most potent compound among the tested compound from above described was evaluated for in vitro short term anticancer study by Dalton’s Lymphoma Ascites (DLA). An effect of acridine derivatives on DNA synthesis raised the hypothesis that acridine moieties secondary effect on biochemical pathways, including protein and lipid metabolism exists, which suggest acridine derivatives could be considered multi targets drugs.Structural modification may lead to synthesize more acridine derivatives. The present study opens new vista for DNA intercalating anticancer compounds and their further in vivo investigation.