Biological activity and DNA sequence specificity of synthetic carbamoyl analogues of distamycin

A new penta(N-methylpyrrole carboxamide) analogue of the antibiotic distamycin has been synthesized in which the N-terminal formylamino group was replaced by a carbamoyl moiety It was substantially more stable than distamycin in aqueous solution and bound to DNA with about the same affinity constant. It had an exemplary margin of selectivity against herpes simplex virus type 1-infected HEp-2 cells in culture compared to uninfected control cells, and was equipotent with distamycin. For comparison, data for analogues containing fewer N-methylpyrrole carboxamide units and/or lacking the carbamoyl replacement are presented. Extensive DNase I footprinting experiments were conducted and revealed that all the distamycin analogues bound to AT-rich nucleotide sequences in three different restriction fragments, irrespective of how many pyrrole rings or which terminal moiety they contained. However, the relative strength of footprints differed significantly among the various compounds, though the apparent size of the binding site did not. With semi-synthetic DNA containing inosine and 2,6-diaminopurine residues in place of guanosine and adenine, respectively, the compounds recognized new binding sites composed of IC-rich clusters and were excluded from binding to their canonical sites. This showed that the process of specific sequence recognition was critically dominated by the placement of the purine 2-amino group in the minor groove of the double helix.