On The Formation of Oxygenated Radicals by Fredericamycin A and Implications to its Anticancer Activity: An ESR Investigation

"It has been recently suggested that the exceptionally high antitumor and antibacterial activity of natural fredericamycin A (FMA) is related to its ability to spontaneously generate the superoxide anion (02-) and hydroxyl (""OH) radicals in aerobic solutions [Hilton, B. D., Misra, R., & Zweier, J. L. (1986) Biochemistry 25, 5533]. With a view to understand the mechanistic details, attempts were made to reproduce earlier electron spin resonance (ESR) evidence for the oxygenated free radical formation in well-aerated solutions of natural FMA in dimethyl sulfoxide and dilute H 20 2• Little or no evidence was obtained for the formation of the 0 2- and methoxy (""OCH3) radicals, while the detected formation of the 'OH and methyl ('CH3) radicals was attributable largely to mechanisms not involving FMA. These results thus reopen the T question regarding the mechanism of its exceptionally high tumoricidal-bacteriocidal activity. This paper reports on an electron spin resonance (ESR) investigation of the formation of oxygenated free radicals in solutions of natural fredericamycin A (abbreviated as FMA) which has recently been reported to be a highly potent anti¬tumor antibiotic (Pandey et al., 1981; Warnick-Pickle et al., 1981). The impetus for this undertaking was provided by recent reports (Hilton et al., 1986; Stinson, 1986) suggesting that the strong tumoricidal activity with high therapeutic—toxic index of FMA was related to its ability for the spontaneous generation of superoxide anion (02-) and hydroxyl (OH) radicals. For example, the suggested mechanism for the formation of 02- by FMA was (Hilton et al., 1986)FMA-OH + O2 FMA-O + H+ + 02-Here FMA—OH is simply the FMA molecule emphasizing the —OH moiety which becomes FMA-O, the free radical rep¬resenting the oxidized form of the drug. While some other highly effective anticancer drugs in current clinical application, such as bleomycin, are also thought to exert their antitumor effects via the formation of reactive drug-based or oxygenated free radicals (Carmichael et al., 1985; Gianni et al., 1983), FMA was considered unique in that only in this case the free radical formation was thought to be spontaneous, rather than through a metal- or an enzyme-activated process (Hilton et al., 1986; Stinson, 1986). Thus it was suggested (Hilton et al., 1986; Stinson, 1986) that FMA could be the forerunner of a new class of anticancer drugs. However, we noted that while the hypothesis seemed plausible, the reported ESR spectral evidence appeared to be ambiguous. Because of the high significance of the earlier conclusions (Stinson, 1986), we undertook a systematic ESR study of the radical formation in FMA solutions, and as discussed below, our conclusions disagree with those reported earlier."