COMPARATIVE METABOLISM OF PHENANTHRO[3,4-B]THIOPHENE, PHENANTHRO[4,3-B]THIOPHENE AND THEIR CARBON ANALOG BENZO[C]PHENANTHRENE BY RAT LIVER MICROSOMES

Abstract

Phenanthro[3,4-b]thiophene (P[3,4-b]T) and phenanthro[4,3-b]thiophene (P[4,3-b]T) are thiasters of weakly mutagenic benzo[c]phenanthrene (B[c]P). These polycyclic sulfur heterocycles (thia-PAHs) represent a group of chemicals which have been identified in cigarette smoke. P[3,4-b]T is a potent mutagen in Salmonella typhimurium strain TA100 in the presence of rat liver S9, whereas its isosteric isomer P[4,3-b]T is a nonmutagenic compound. In order to understand the mechanism underlying the differences in the mutagenic activity of P[3,4-b]T and P[4,3-b]T, we have investigated the metabolism of P[3,4-b]T, P[4,3-b]T, and their carbon analogue B[c]P by rat liver microsomes. The liver microsomes from rats treated with Aroclor 1254 metabolized P[3,4-b]T, P[3,4-b]T, and B[c]P at a rate nearly 7- to 9-fold greater than of the control liver microsomes. High-performance liquid chromatography (HPLC) analysis of the metabolites formed showed that B[c]P was metabolized almost exclusively to its dihydrodiols which comprised predominantly K-region diol as noted in the previous studies. Our preliminary studies on the metabolism of P[3,4-b]T, P[4,3-b]T and B[c]P by liver microsomes from control and Aroclor 1254-treated rats have shown a significant reduction in the formation of 6,7-diol (K-region diol) and 8,9-diol (diol with a bay-region double bond) of the two thia-PAHs compared to the formation of analogous 5,6-diol (K-region diol) and 3,4-diol (diol with a bay-region double bond) from B[c]P. Both P[3,4-b]T and P[4,3b]T produced a major, relatively nonpolar metabolite(s) (80–96% of total metabolites). These studies indicate that the highly mutagenic P[3,4-b]T is not metabolized to dihydrodiol with a bay-region double bond to any greater extent than the weakly or nonmutagenic B[c]P or P[4,3-b]T, suggesting that the metabolite(s) other than P[3,4-b]T8,9-diol is likely to be involved in the mutagenicity of P[3,4-b]T.

Keywords:

• benzo[c]phenanthrene
• metabolism
• phenanthro[3,4-b]thiophene
• phenanthro[4,3-b]thiophene
• rat liver microsomes

Acknowledgments

This research was supported by a grant awarded by Philip Moris U.S.A., Inc.

Notes

^a Values are means of duplicate determinations and represent the percentage of total radioactivity which emerges from the column prior to [G-³H]-BPT.

^b Unknown A represents total unidentifiable metabolites other than B that elutes as a single major peak at ∼ 46–47 min; peak B coelutes with one of the products produced by the treatment of P[3,4-b]T with H₂O₂/TFA¹⁵, or m-CPBA¹⁶ (data not shown).

^c Unknown A represents total unidentifiable metabolites other than B which elutes as a single major peak; peak B coelutes with 10-hydroxy-P[4,3-b]T⁹ but not with the product obtained after treating P[4,3-b]T with H₂O₂/TFA (data not shown).