Electrochemical and electron spin resonance studies of actinomycin D and other phenoxazones

Electrochemical studies on actinomycin D (1) and two analogs, 2-amino-3-phenoxazone (2) and 1,2,4-trichloro-7-nitrophenoxazone (3) were analyzed by polarography and ESR spectroscopy. The polarograms of the three compounds in acetonitrile all show two reduction waves. ESR experiments confirm that the first reduction wave corresponds to a one-electron transfer process which produces a phenoxazone free radical anion and the second wave corresponds to a subsequent one-electron transfer producing a diamagnetic dianion. Substitution with electron-withdrawing groups such as NO2 (at C-7) and chloro (at C-1, C-2 and C-4)3 facilitated the reduction of the phenoxazone ring system to a free radical (i.e., half-wave potentials; 1, -0.815 V; 2, -0.920 V; 3, -0.135 V). It was found, by computer simulation of the ESR spectra, that the spin density in the electrochemically generated free radicals from 1, 2 and 3 was preferentially located in the benzenoid ring and at the N-10 nitrogen. For radicals obtained from 1 and 2, only a small residual spin density could be detected in the quinoid ring. Since 1 can be metabolized to a free radical in cells, these free radical forms of 1 and its analogs may represent reactive forms of the phenoxazone nucleus.