The metabolic sulfonation and side-chain oxidation of 3′-hydroxyisosafrole in the mouse and its inactivity as a hepatocarcinogen relative to 1′-hydroxysafrole

The chemically synthesized sulfuric acid esters of 1'-hydroxysafrole and 3'-hydroxyisosafrole, 1'-sulfooxysafrole and 3'-sulfooxyisosafrole, respectively, are both strong electrophiles. Each ester reacted with deoxyguanosine (dGuo) in aqueous solution to form both safrol-1'-yl- and isosafrol-3'-yl-deoxyguanosine adducts. Both 1'-hydroxysafrole and 3'-hydroxyisosafrole were also formed from each ester in the presence of water. When either 1'-[3H]hydroxysafrole or 3'-[3H]hydroxyisosafrole was incubated with mouse liver cytosols fortified with 3'-phosphoadenosine-5'-phosphosulfate (PAPS) and RNA, similar levels of RNA- and protein-bound adducts were formed; thus, the hepatic sulfotransferase activities for these two substrates appear to be similar. In contrast, the levels of hepatic nucleic acid and protein adducts formed after administration of 3'-[3H]hydroxyisosafrole to mice were only 2-4% and 8-14%, respectively, of those obtained after an equimolar dose of 1'-[3H]hydroxysafrole. Likewise, when 3'-hydroxyisosafrole was injected into 12-day-old male B6C3F1 mice at a level of 0.1 or 2.5 mumol/g body wt., the average numbers of hepatomas per mouse (0.2 and 0.4, respectively) were not significantly increased over the average number for mice treated only with the solvent (0.2). By contrast, mice that received 0.1 mumol of 1'-hydroxysafrole/g body wt. developed about 2 hepatomas per mouse. The metabolism of 3'-hydroxyisosafrole in the rat and mouse differed markedly from that of 1'-hydroxysafrole. 3'-Hydroxyisosafrole rapidly underwent side-chain oxidation to yield 3,4-methylenedioxycinnamic acid and 3,4-methylenedioxybenzoic acid. In the first 4 h, 3,4-methylenedioxybenzoyl glycine and 3,4-methylenedioxycinnamoyl glycine, the major urinary metabolites, together accounted for 39% and 63% of the dose administered to rats and mice, respectively. The glucuronide of 3'-hydroxyisosafrole was not detected in the urine, whereas urinary excretion of the glucuronide of 1'-hydroxysafrole at 2 h accounted for approx. 40% of a dose of 1'-hydroxysafrole.