Role of cytochrome P-450 2E1 in methacrylonitrile metabolism and disposition.
Methacrylonitrile (MAN) is a widely used aliphatic nitrile and is structurally similar to the known rat carcinogen and suspected human carcinogen acrylonitrile (AN). There is evidence that AN is metabolized via the cytochrome P-450 (CYP) 2E1. Recently, we identified two biliary conjugates originating from the interaction of MAN and its epoxide with glutathione. Mercapturic acids formed via the degradation of the two conjugates were also identified in rat and mouse urine. Additionally, a significant portion of MAN was eliminated in the expired air as CO2 (formed via the epoxide pathway) and unchanged MAN. The objective of the present work was to determine whether CYP2E1 is involved in the oxidative metabolism of MAN as was suggested for AN. 2-14C-MAN was administered to CYP2E1-null or wild-type mice by gavage at 12 mg/kg. Although total urinary and fecal excretion of MAN-derived radioactivity was slightly different in CYP2E1-null versus wild-type mice, the ratio of mercapturic acids originating from the epoxide-glutathione versus MAN-glutathione conjugates were lower in urine of CYP2E1-null mice than in that of wild-type animals. Exhalation of MAN-derived organic volatiles (primarily parent MAN) was 12- and 42-fold greater in female and male CYP2E1-null mice than in wild-type mice, respectively. Additionally, exhalation of CO2 derived from metabolism of MAN via the CYP2E1 pathway was 3- to 5-fold greater in wild-type than in CYP2E1-null animals. Although these data indicate that CYP2E1 is the principal enzyme responsible for the oxidative metabolism of MAN, other cytochrome P-450 enzymes may be involved. Assessment of MAN metabolism in CYP2E1-null mice pretreated with 1-aminobenzotriazole (CYP inhibitor) resulted in a further decrease in oxidative metabolites of MAN. Comparison of the tissue concentrations of MAN-derived radioactivity in mouse tissues revealed that MAN-derived radioactivity is generally higher in wild-type > CYP2E1-null mice > CYP2E1-null mice pretreated with 1-aminobenzotriazole, suggesting a direct relationship between MAN oxidative metabolism and the half-life of MAN and/or its metabolites in various tissues. It is therefore concluded that MAN oxidative metabolites such as the epoxide intermediate have greater reactivity than parent MAN.
Ghanayem
B I
BI
National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA. Ghanayem@niehs.nih.gov
Sanders
J M
JM
Chanas
B
B
Burka
L T
LT
Gonzalez
F J
FJ
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000652:urine; D000818:Animals; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism*; D005243:Feces / Q000737:chemistry; D005260:Female; D008297:Male; D008689:Methacrylates / Q000378:metabolism / Q000493:pharmacokinetics* / Q000633:toxicity; D051379:Mice; D018345:Mice, Knockout; D009570:Nitriles / Q000378:metabolism / Q000493:pharmacokinetics* / Q000633:toxicity; D010084:Oxidation-Reduction; D014018:Tissue Distribution
D008689:Methacrylates; D009570:Nitriles; C008385:methacrylonitrile; C054605:N-acetyl-S-(2-carboxypropyl)cysteine; D019392:Cytochrome P-450 CYP2E1; D000111:Acetylcysteine