Mitomycin C linked to DNA minor groove binding agents: synthesis, reductive activation, DNA binding and cross-linking...

Mitomycin C (MC) is a natural cytotoxic agent used in clinical anticancer chemotherapy. Its antitumor target appears to be DNA. Upon bioreductive activation MC alkylates and cross-links DNA. MC derivatives were synthesized in which MC was linked to DNA minor groove binding agents, analogous to netropsin and distamycin. One, two and three N-methylpyrrole carboxamide units were conjugated with MC by a (CH2)5-tether to the 7-amino group of MC (11, 12 and 13, respectively). In contrast to MC 11, 12 and 13 displayed non-covalent affinity to DNA. Their bioreductive activation by NADPH-cytochrome c reductase proceeded as fast as that of MC. Metabolites arising from reductive and low-pH activation were characterized and found to be analogous to those of MC. DNA cross-linking activities were weak and decreased with an increasing number of N-methylpyrrole carboxamide units linked with the mitomycin molecule. No adducts were formed with calf thymus DNA in detectable amounts. In vitro antitumor activities of 11-13 were determined using the NCI in vitro antitumor screen. The conjugates 11-13 are growth inhibitory; however, their activities are 1.5-2 orders of magnitude lower than that of MC. COMPARE analysis indicates that the mechanism of the action of 11 and 12 correlates moderately with MC but negatively with distamycin. Conjugate 13 correlates neither with MC nor with distamycin. The results suggest that the basic cause of the observed low activity of the MC-minor groove binder conjugates is the fast irreversible decay of the activated MC, competing effectively with the slow drug delivery to CpG sites, required for the alkylation.