title: An evaluation of the mutagenicity, metabolism, and DNA adduct formation of 5-nitrobenzo[b]naphtho[2,1-d]thiophene.
abstract: Thioarenes, sulfur-containing polycyclic aromatic compounds, are environmental contaminants suspected of posing human health risks. In this study, 5-nitrobenzo[b]naphtho[2,1-d]thiophene (5-nitro-BNT), a nitrated-thioarene, was examined for its mutagenicity, metabolism and subsequent formation of DNA adducts. 5-Nitro-BNT was weakly mutagenic in Salmonella typhimurium strains TA98 and TA100 without Aroclor-1254-induced rat liver S9 (S9), and its activity was increased in the presence of S9. Anaerobic metabolism of 5-nitro-BNT by S9 or xanthine oxidase (XO) produced one major metabolite, identified as 5-amino-BNT by NMR, MS, and UV 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 trans-9,10-dihydroxy-9,10-dihydro-5-amino-BNT (5-amino-BNT-9,10-diol). DNA adduct analyses were performed using the (32)P-postlabeling assay and reversed-phase HPLC. Three major XO-derived calf thymus DNA adducts were detected. On the basis of their chromatographic mobilities, two adducts were identified as reaction products of 5-nitro-BNT with 2'-deoxyguanosine and one adduct 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 chromatographic mobilities, two were identified as reaction products of 5-nitro-BNT with 2'-deoxyguanosine and one with 2'-deoxyadenosine. Incorporation of 1-aminobenzotriazole (a P450 inhibitor) in the incubation mixture resulted in a loss of these adducts, confirming enzymatic mediation by P450. Aerobic 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 as observed with 5-nitro-BNT and 5-nitro-BNT-9,10-diol. These results provide additional information that both ring oxidation and nitroreduction are involved in the metabolism, DNA adduct formation and mutagenicity of 5-nitro-BNT.
authors: U.S. Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Environmental Carcinogenesis Division, MD-68, Research Triangle Park, North Carolina 27711, USA.
authors: L C
authors: LC
authors: King
authors: M J
authors: MJ
authors: Kohan
authors: L
authors: L
authors: Brooks
authors: G B
authors: GB
authors: Nelson
authors: J A
authors: JA
authors: Ross
authors: J
authors: J
authors: Allison
authors: L
authors: L
authors: Adams
authors: D
authors: D
authors: Desai
authors: S
authors: S
authors: Amin
authors: W
authors: W
authors: Padgett
authors: G R
authors: GR
authors: Lambert
authors: A M
authors: AM
authors: Richard
authors: S
authors: S
authors: Nesnow
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D018736:DNA Adducts; D004785:Environmental Pollutants / Q000009:adverse effects* / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D009152:Mutagenicity Tests; D010084:Oxidation-Reduction; D011084:Polycyclic Aromatic Hydrocarbons / Q000009:adverse effects* / Q000378:metabolism; D051381:Rats; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics*; D013876:Thiophenes / Q000009:adverse effects* / Q000378:metabolism; D013950:Thymus Gland / Q000187:drug effects
chemical_list: D018736:DNA Adducts; D004785:Environmental Pollutants; D011084:Polycyclic Aromatic Hydrocarbons; D013876:Thiophenes; C050001:benzo(b)naphtho(2,1-d)thiophene