An evaluation of the mutagenicity, metabolism, and DNA adduct formation of5-nitrobenzo[b]naphtho[2,1-d]thiophene

Thioarenes, sulfur-containing polycyclic aromatic compounds, are environmen tal contaminants suspected of posing human health risks. In this study, 5-n itrobenzo [b]naphtho[2,1-d]thiophene (5-nitro-BNT), a nitrated-thioarene, w as examined for its mutagenicity, metabolism and subsequent formation of DN A adducts. 5-Nitro-BNT was weakly mutagenic in Salmonella typhimurium strai ns TA98 and TA100 without Aroclor-1254-induced rat liver S9 (S9), and its a ctivity was increased in the presence of S9. Anaerobic metabolism of 5-nitr o-BNT by S9 or xanthine oxidase (XO) produced one major metabolite, identif ied as 5-amino-BNT by NMR, MS, and W spectroscopy and by comparison with an authentic standard. Aerobic S9 metabolism of 5-nitro-BNT produced a major metabolite, identified as trans-9,10-dihydroxy-9, 10-dihydro-5-nitro-BNT (5 -nitro-BNT-9,10-diol). Also present was a minor amount of 5-amino-BNT and t rans-9,10-dihydroxy 9,10-dihydro-5-amino-BNT (5-amino-BNT-9,10-diol). DNA a dduct analyses were performed using the P-32-postlabeling assay and reverse d-phase HPLC. Three major XO-derived calf thymus DNA adducts were detected. On the basis of their chromatographic mobilities, two adducts were identif ied as reaction products of 5-nitro-BNT with 2 ' -deoxyguanosine and one ad duct with 2 ' -deoxyadenosine. Incorporation of allopurinol (a specific XO inhibitor) in the incubation mixture resulted in loss of all three adducts, confirming enzymatic mediation by XO. Aerobic S9 activation of 5-nitro-BNT with calf thymus DNA produced three adducts. On the basis of their chromat ographic mobilities, two were identified as reaction products of 5-nitro-BN T with 2 ' -deoxyguanosine and one with 2 ' -deoxyadenosine. Incorporation of 1-aminobenzotriazole (a P450 inhibitor) in the incubation mixture result ed in a loss of these adducts, confirming enzymatic mediation by P450. Aero bic S9-catalyzed metabolism of 5-nitro-BNT-9, 10-diol produced the same DNA adducts as observed with 5-nitro-BNT. Aerobic S9-catalyzed metabolism of 5 -amino-BNT-9,10-diol produced the same deoxyadenosine-derived DNA adducts a s observed with 5-nitro-BNT and 5-nitro-BNT-9,10-diol. These results provid e additional information that both ring oxidation and nitroreduction are in volved in the metabolism, DNA adduct formation and mutagenicity of 5-nitro- BNT.