Ternary Copper Complexes for Photocleavage of DNA by Red Light: Direct Evidence for Sulfur-to-Copper Charge Transfer and d-d Band Involvement

A new class of ternary copper(II) complexes of formulation $[Cu(L^n)B](ClO_4)$ (1-4), where $HL^n$ is a NSO-donor Schiff base $(HL^1, HL^2)$ and B is a NN-donor heterocyclic base viz. 1,10-phenanthroline (phen) and 2,9-dimethyl-1,10-phenanthroline (dmp), are prepared, structurally characterized, and their DNA binding and photocleavage activities studied in the presence of red light. Ternary complex $[Cu(L^3)(phen)](ClO_4)$ (5) containing an ONO-donor Schiff base and a binary complex $[Cu(L^2)_2]$ (6) are also prepared and structurally characterized for mechanistic investigations of the DNA cleavage reactions. While 1-4 have a square pyramidal (4 + 1) $CuN_3OS$ coordination geometry with the Schiff base bonded at the equatorial sites, 5 has a square pyramidal (4 + 1) geometry with $CuN_3O_2$ coordination with the alcoholic oxygen at the axial site, and 6 has a square planar trans-$CuN_2O_2$ geometry. Binding of the complexes 1-4 to calf thymus DNA shows the relative order: phen >> dmp. Mechanistic investigations using distamycin reveal minor groove binding for the complexes. The phen complexes containing the Schiff base with a thiomethyl or thiophenyl moiety show red light induced photocleavage. The dmp complexes are essentially photonuclease inactive. Complexes 5 and 6 are cleavage inactive under similar photolytic conditions. A $10 \mu M$ solution of 1 displays a 72% cleavage of SC DNA $(0.5 \mu g)$ on an exposure of 30 min using a 603 nm Nd:YAG pulsed laser (60 mJ/P) in Tris-HCl buffer (pH 7.2). Significant cleavage of 1 is also observed at 694 nm using a Ruby laser. Complex 1 is cleavage inactive under argon or nitrogen atmosphere. It shows a more enhanced cleavage in pure oxygen than in air. Enhancement of cleavage in $D_2O$ and inhibition with sodium azide addition indicate the possibility of the formation of singlet oxygen as a reactive intermediate leading to DNA cleavage. The d-d band excitation with red light shows significant enhancement of cleavage yield. The results indicate that the phen ligand is necessary for DNA binding of the complex. Both the sulfur-to-copper charge transfer band and copper d-d band excitations helped the DNA cleavage. While the absorption of a red photon induces a metal d-d transition, excitation at shorter visible wavelengths leads to the sulfur-to-copper charge transfer band excitation at the initial step of photocleavage. The excitation energy is subsequently transferred to ground state oxygen molecules to produce singlet oxygen that cleaves the DNA