Effect of ancillary ligands on the DNA interaction of [Cr(diimine)3]3+ complexes containing the intercalating dipyridophenazine ligand.

The synthesis of photoluminescent Cr(III) complexes of the type [Cr(diimine)(2)(DPPZ)](3+) are described, where DPPZ is the intercalating dipyridophenazine ligand, and diimine corresponds to the ancillary ligands bpy, phen, DMP, and TMP (where bpy = 2,2'-bipyridine, phen = 1,10-phenanthroline, DMP = 5,6-dimethyl-1,10-phenanthroline, and TMP = 3,4,7,8-tetramethyl-1,10-phenanthroline). For TMP, DMP, and phen as ancillary ligands, the complexes have also been resolved into their Lambda and Delta optical isomers. A comparison of the photophysical and electrochemical properties reveal similar (2)E(g) --> (4)A(2g) (O(h)) emission wavelengths and lifetimes, and a variation of 110 mV in the (2)E(g) excited state oxidizing power. A detailed investigation has been undertaken of ancillary ligand effects on the DNA binding of these complexes with a range of polynucleotides. For all four complexes, emission is quenched by the addition of calf thymus B-DNA, with the emission lifetime data yielding bimolecular quenching rate constants close to the diffusion controlled limit. Equilibrium dialysis studies have established a general predilection for AT base binding sites, while companion experiments with added distamycin (a selective minor groove binder) provide evidence for a minor groove binding preference. For the case of [Cr(TMP)(2)(DPPZ)](3+), concomitant equilibrium dialysis and circular dichroism measurements have demonstrated very strong enantioselective binding by the Lambda optical isomer. The thermodynamics of DNA binding have also been explored via isothermal titration calorimetry (ITC). The ITC data establish that the primary binding mode for all four Cr(III) complexes is entropically driven, a result that is attributed to the highly favorable free energy contribution associated with the hydrophobic transfer of the Cr(III) complexes from solution into the DNA binding site.

Capillary electrophoresis as a probe of enantiospecific interactions between photoactive transition metal complexes and DNA.

Recently, we have demonstrated the capacity to separate chiral transition metal (TM) complexes of the type [M(diimine)(3)](n+) using CE buffers containing chiral tartrate salts. In separate work, several chromium(III)-tris-diimine complexes in particular have been shown to bind enantioselectively with calf-thymus (CT) DNA, and a qualitative assessment of the relative strength and enantiospecificity of this interaction is of significant interest in the characterization of these complexes as potential DNA photocleavage agents. Here, we describe two convenient approaches to investigate such binding behavior using chiral CE. For complexes with lower DNA affinities exhibiting primarily surface binding, DNA itself is used as the chiral resolving agent in the electrophoretic buffer. In this approach, resolution of the TM complexes into their Lambda and Delta isomers is achieved with the isomer eluting later exhibiting superior binding affinity toward DNA. For more strongly bound TM complexes containing ligands known to intercalate with DNA, the [Cr(diimine)(3)](3+) complexes are preincubated with oligonucleotide and subsequently enantiomerically resolved in a dibenzoyl-L-tartrate buffer system that facilitates analysis of the unbound TM species only. Differences in isomer binding affinity are distinguished by the relative peak areas of the Lambda- and Delta-isomers, and relative binding strengths of different complexes can be inferred from comparison of the total amount of unbound complex at equivalent DNA/TM ratios.