The interaction of DNA molecule with various actinocine derivatives, which are analogues of the antibiotic actinomycin D, was investigated by spectral, hydrodynamic and optical methods. The thermodynamic parameters of the interaction and the stoichiometry of the complexes were determined by spectrophotometric titration. A mode of binding and the structure of the complexes were determined by analyzing the changes in the intrinsic viscosity and the optical anisotropy of the macromolecule upon complexation. The dependence of thermodynamic parameters and mode of binding with DNA of compounds The dependence of thermodynamic parameters and mode of binding of the actinocine derivatives with DNA on substitutes’ nature and ion strength of a medium was studied. It was shown that protonization of substitute of the chromophore gives rise to compaund affinity to DNA. The intercalation of the actinocin chromophore in the DNA double helix occurs only under optimal linker’s length which binding the chromophore with protonized group of the substitute. In the case of crown-containing derivatives of actinocine, the nature of the counterions and the ionic strength of a medium affect not only on the affinity of compounds to DNA, but on its mode of binding with the macromolecule.