A series of methoxy- and fluorine-substituted [salophene]platinum(II) complexes (salophene=N,N'-bis(salicylidene)-1,2-phenylenediamine) were synthesized and characterized by (1) H NMR spectroscopy and mass spectrometry. The structure was confirmed on the example of [5-OCH3 -salophene]platinum(II) (4-Pt) by crystal structure analysis. The cytotoxicity of all complexes against MCF-7 cells showed strong dependence on the kind of substituent and its position on the salicylidene moiety, whereas 1-Pt (H), 3-Pt (4-OCH3 ), and 6-Pt (3-F) exhibited high antiproliferative effects (IC50 <2 μM). Drug lipophilicity and cellular accumulation were analyzed in an attempt to explain the differences in antitumor potency. To gain insight into their mode of action, DNA interaction studies were performed, in which compounds such as 1-Pt acted as non-DNA-binding platinum anticancer drugs, as neither intercalation nor DNA covalent binding were detected.