RuII (η 6 -arene) complexes, especially with bioactive ligands, are considered as very promising compounds for anticancer drug design. We have shown recently that Ru II (η 6 -p-cymene) complexes with 3-hydroxyflavone ligands exhibit very high in vitro cytotoxic activities correlating with a strong inhibition of topoisomerase IIα. In order to expand the structure-activity relationships and to determine the impact 10 of lipophilicity of the arene ligand and of the hydrolysis rate on anticancer activity, a series of novel 3-hydroxyflavone derived Ru II (η 6 -arene) complexes were synthesised. Furthermore, the impact of the heteroatom in the bioactive ligand backbone was studied by comparing the cytotoxic activity of Ru II (η 6 -pcymene) complexes of 3-hydroxyquinolinone ligands with that of their 3-hydroxyflavone analogues. To better understand the behaviour of these Ru II complexes in aqueous solution, the stability constants and 15 pK a values for complexes and corresponding ligands were determined. Furthermore, the interaction with the DNA model 5'-GMP and with a series of amino acids was studied in order to elucidate potential biological target structures.