SULFURIC-ACID ESTERS AS MAJOR ULTIMATE ELECTROPHILIC AND HEPATOCARCINOGENIC METABOLITES OF 4-AMINOAZOBENZENE AND ITS N-METHYL DERIVATIVES IN INFANT MALE C57BL/6JXC3H/HEJF1 (B6C3F1) MICE

Liver cytosols from 12-day-old male C57BL/6 .times. C3H/HeJ F1 (B6C3F1) mice contain 3''-phosphoadenosine-5''-phosphosulfate (PAPS)-dependent sulfotransferase activity for N-hydroxy-4-aminoazobenzene and N-hydroxy-N-methyl-4-aminoazobenzene. No acetyl co-enzyme A-dependent transacetylase activity for these hydroxylamines was detected in the cytosols. Pentachlorophenol (PCP) and 2,6-dichloro-4-nitrophenol were only moderately active inhibitors of the sulfotransferase activity; at a 100-.mu.M concentration each compound inhibited the activity by only 50-80%. A single dose of 0.04 .mu.mol/g body weight of PCP administered to 12-day-old male B6C3F1 mice 45 min prior to a single dose of 0.1 .mu.mol/g body weight of [3H]4-aminoazobenzene ([3H]AB) or [3H]N,N-dimethyl-4-aminoazobenzene ([3H]DAB) inhibited DNA adduct formation by .apprx. 50%. Under identical conditions, PCP also reduced the average number of hepatomas induced per mouse at 9 months by AB and N-methyl-4-aminoazobenzene (MAB) by 52 and 36%, respectively. PCP strongly inhibited the hepatocarcinogenicity of DAB or AB when this agent was administered in the diet with either dye to female CD-1 mice over a 10 month period. Single doses of 0.15 .mu.mol/g body weight of [3H]AB and [3H]DAB bound to hepatic DNA of 12-day-old brachymorphic B6C3F2 mice, which are deficient in the synthesis of PAPS, at levels 15 and 20%, respectively, of those found in their phenotypically normal litter mates. Under identical conditions, the incidence of hepatomas in brachymorphic mice at 9 months were 11 and 29%, with averages of 0.2 and 0.8 hepatomas/mouse for AB and MAB, respectively. Incidences of 77 and 86%, with averages of 6.6 and 5.4 hepatomas/mouse, respectively, were found in their phenotypically normal litter mates. These data strongly indicate that N-sulfooxy-AB is a major ultimate electrophilic and hepatocarcinogenic metabolite of AB in mice. Similarly, this ester and N-sulfooxy-N-methyl-4-aminoazobenzene appear to be critical metabolites for these activities of DAB and MAB.