Cancer is a leading cause of death worldwide, currently the cancer treatments are radiation therapy, surgery or chemotherapy with the well-known side effects. So far there is no curative therapy available for most of disseminated cancer and thereby the development and discovery of novel therapeutic agents are largely needed. From the development of cisplatin, many scientific groups are actively worked/working for the inorganic anticancer drugs. and have developed a number of Ru(II) complexes with lower systemic toxicity. Ruthenium complexes can penetrate effectively with the tumor cells and bind with DNA. In view of this, we have synthesized novel Ru(II) isatin complex cis-[Ru II (Phen)2 FPIMI] ClO4.2H2O from the reactions between cis-[Ru (Phen)2 Cl2].2H2O and 4fluoro phenyl imino methyl isatin(FPIMI). The new complex was analyzed by elemental analysis and spectral (UV-Vis, FTIR, and 1 H NMR) studies. Further, the complex was subjected to DNA intercalation studies, in vitro anticancer studies and cytotoxic activities. DNA intercalative assay was followed by UV-Vis spectral titration studies with relatively high DNA binding constant, Kb=4.1±2 10 5 M -1 at room temperature. The above studies showed intercalative mode of cleavage with Calf thymus(CT) DNA. The newly synthesized ruthenium(II) complex exhibited more effective in vitro cytotoxic activity against selected cell lines with IC50 value 26±0.5 µM. Also, the anticancer activity of the compound is apparently four times more potent than one of the presently used clinical drug Cisplatin against human tumor cell lines with less cytotoxicity.