Kinetics of Trihalogenated Acetic Acid Metabolism and Isoform Specificity in Liver Microsomes

• Published in: International journal of toxicology 2011-10, Vol.30 (5), p.551-561
• Main Authors: Saghir, Shakil A., Ghanayem, Burhan I., Schultz, Irvin R.
• Format: Article
• Language: English
• Subjects: Acetates - pharmacokinetics; Acetates - toxicity; Animals; Chloroacetates; Cytochrome P-450 CYP2E1 - metabolism; Cytochrome P-450 Enzyme System - metabolism; Disinfection; Drinking Water - analysis; Halogenation; Humans; Hydrocarbons, Brominated; Male; Mice; Mice, Knockout; Microsomes, Liver - drug effects; Microsomes, Liver - metabolism; NADPH-Ferrihemoprotein Reductase - metabolism; Protein Isoforms - metabolism; Rats; Rats, Inbred F344; Trichloroacetic Acid - pharmacokinetics; Trichloroacetic Acid - toxicity
• ISSN: 1091-5818; 1092-874X
• DOI: 10.1177/1091581811414213

This study determined the metabolism of 3 drinking water disinfection by-products (halogenated acetic acids [HAAs]), bromodichloroacetic acid (BDCAA), chlorodibromoacetic acid (CDBAA), and tribromoacetic acid (TBAA), using rat, mouse, human liver microsomes, and recombinant P450. Metabolism proceeded by reductive debromination forming a di-HAA; the highest under nitrogen >>2% oxygen > atmospheric headspaces. V max for the loss of tri-HAA was 4 to 5 times higher under nitrogen than atmospheric headspace. Intrinsic metabolic clearance was TBAA>CDBAA>>BDCAA. At the high substrate concentrations, tri-HAA consumption rate was 2 to 3 times higher than the formation of di-HAA. Liberation of Br− from TBAA corresponded to the expected amount produced after DBAA formation, indicating retention of Br− by additional metabolite/metabolites. Subsequent experiments with CDBAA detected negligible formation of chlorodibromomethane (CDBM) and failed to account for the missing tri-HAA. Carbon monoxide and especially diphenyleneiodonium ([DPI] P450 reductase inhibitor) blocked CDBAA metabolism. Other chemical inhibitors were only partially able to block CDBAA metabolism. Most effective were inhibitors of CYP 2E1 and CYP 3A4. Immunoinhibition studies using human liver microsomes and anti-human CYP 2E1 antibodies were successful in reducing CDBAA metabolism. However, CDBAA metabolism in wild-type (WT) and CYP 2E1 knockout (KO) mouse liver microsomes was similar, suggesting significant interspecies differences in CYP isoform in tri-HAA metabolism. Additional assessment of CYP isoform involvement was complicated by the finding that recombinantly expressed rat and human P450 reductase was able to metabolize CDBAA, which may be a contributing factor in interspecies differences in tri-HAA metabolism.