Kinetic and equilibrium studies on the polyazamacrocycle neotetren:: metal-complex formation and DNA interaction

The macrocyclic polyamine 2,5,8,11,14-pentaaza[15]-[15](2,9)[1,10] phenanthrolinophane (neotetren) is studied in its ability to coordinate Cu(II) even at very low pH values and to interact, as a metal complex, with DNA. The kinetics and equilibria for 1 : 1 and 2 : 1 metal - ligand complexes formation are studied by the stopped-flow method and UV spectrophotometry. Differently protonated complexes are formed, with rate constants much lower than that of water exchange at copper( II) and other Cu( II)/amine systems, this behaviour being ascribed to ring effects and intra-molecular hydrogen bonds. Concerning the DNA/copper(II) - neotetren complexes interaction, analysis of data suggests an intercalative mode of binding. The kinetic results for both DNA/CuL and DNA/Cu2L systems agree with the sequence D + S reversible arrow D, S reversible arrow DS where the metal complexes (D) react with the DNA sites (S) leading to fast formation of an externally bound form ( D, S) which is converted into an intercalated complex ( DS). A very slow process is also detected and ascribed to a conformational change in the polynucleotide secondary structure where the metal centre plays a crucial role. Chromatographic experiments demonstrate that both the investigated Cu(II)/ L complexes are able to cleave DNA, but only in the presence of hydrogen peroxide.