Studies Related to the Antitumor Antibiotic Mitomycin B and Models
The base-catalyzed methanolysis of model monofunctional indoles was studied in order to gain information about the relative reactivity of 2- and 3-indolylcarbinyl systems, analogous to the proposed reactive sites of mito- sanes which have been reductively activated. The leaving groups studied were also analogous to those found in the mitomycins. Reaction kinetics were studied by the method of nmr spectroscopy at a number of temperatures, thereby allowing the calculation of activation parameters. The interaction of mitomycin B with DNA was examined using rapid and convenient ethidium fluorescence assays. Studies on the pH dependence of covalent cross- linking and alkylation of DNA by mitomycin B and its azir- idine-ring-opened derivative have confirmed the involvement of both the aziridine and carbamate groups in the covalent attachment to DNA. In conjunction with the pH studies the pKa of mitomycin B was determined. By using a number of DNA's and variations of the ethidium assay it has been possible to partially define the DNA sites involved in covalent attachment. It was found that mitomycin B induces single strand scission in DNA by the generation of superoxide and hydroxyl radicals; this scission is also produced by mitomycin B and mitomycin C when they are covalently bound to the DNA. The effect of intercalated ethidium on the scission produced by mitomycin B and mitomycin C was also investi-gated. Electroanalytical experiments with mitomycin B and its aziridine-ring-opened derivative yielded results consistent with the comparative reactivities found for mitomycin B and mitomycin C.