Two pairs of iron(II) chiral enantiomers fac-Δ-[Fe(S-L1) 3 ][ClO 4 ] 2 and fac-Λ-[Fe(R-L1) 3 ][ClO 4 ] 2 (Δ-1 and Λ-1), fac-Δ-[Fe(S-L2) 3 ][ClO 4 ] 2 and fac-Λ-[Fe(R-L2) 3 ][ClO 4 ] 2 (Δ-2 and Λ-2), L1=(R/S)-(±)-1-naphthyl-N-(pyridine-2-ylmethylene)ethanamine, L2=(R/S)-(±)-2-naphthyl-N-(pyridine-2-ylmethylene)ethanamine were synthesized and characterized by elemental analysis, IR, UV–Vis, CD and 1 H NMR spectra. The X-ray structural analyses of Λ-1 and Δ-2 revealed that the iron(II) complexes possess octahedral coordination geometry for N6 donor atoms by three bidentate ligands. R-L1 ligand induces the fac-Λ isomer, while S-L2 ligand induces the fac-Δ isomer. The enantioselective binding of iron(II) chiral enantiomers to calf-thymus DNA (ct-DNA) has been investigated by methods of UV–Vis, fluorescence, and circular dichroism spectrometry. All the complexes could bind to ct-DNA and showed different binding affinities with the binding constants ranging from 0.91×10 5 to 1.43×10 5 M −1 . Moreover, the Δ enantiomers exhibited more efficient DNA interaction with respect to the Λ enantiomers.