Contribution of CYP2E1 to N-methyl-2-pyrrolidone metabolism.
The involvement of cytochrome P450 2E1 (CYP2E1) in the metabolism of N-methyl-2-pyrrolidone (NMP) was studied with three experimental approaches: in the rat, in vitro in human microsomes, and in human volunteers. NMP was administered dermally (40 mg/kg) to OFA rats to examine the influence of CYP2E1 inhibition (5 mg/kg diethyldithiocarbamate, DETC, 30 min before) and CYP2E1 induction (after 4 days of fasting). The main NMP metabolite 5-hydroxy- N-methylpyrrolidone (5HNMP) in the urine fractions collected during the following 48 h was analysed by gas chromatography-mass spectrometry. CYP2E1 inhibition led to a statistically significant retardation of 5HNMP excretion in urinary fractions collected during the first 12 h. In the group of fasted rats, a two-fold increase of CYP2E1 activity was observed in comparison with the control group. During the first 6 h after dermal administration of NMP to fasted rats, about 33% of the dose was excreted in urine versus 22% in controls. In vitro, NMP (15 mM) was incubated (up to120 min) with human liver microsomes and the formation of 5HNMP followed Michaelis-Menten kinetics with V(max) of 1.1 nmol/min per mg protein and K(m) of 2.4 mM. The formation of 5HNMP was inhibited by 35% in the presence of a monoclonal antibody against CYP2E1, but not by CYP1A2 antibody. In a dermal application experiment, 12 humans volunteers were exposed by means of a dermal patch to 300 mg NMP; five urine fractions were collected during the 48 h following the onset of application in order to measure the major metabolites 5HNMP and 2-hydroxymethylsuccinimide (2HMSI). Before NMP application, a blood sample was collected for the quantification of CYP2E1 mRNA in peripheral blood lymphocytes (PBLs). The mean dermal absorption of NMP was 67.9%. The highest amount of 5HNMP was excreted in urine in the fraction collected between 6-12 h (12.6% of dose), while 2HMSI peaked in fractions 12-24 h and 36-48 h (3.3 and 3.2% of dose, respectively). A significant relationship was found between CYP2E1 mRNA content in PBLs and the amount of both the metabolites excreted in urine within 24 h ( r(2)=0.54, P<0.01). It is concluded that CYP2E1 is involved in the first steps of NMP metabolism in the rat and, to a lesser extent, in humans. Since large variations in CYP2E1 activity exist in the human population (at least 5-fold range), it seems justified to take into account the activity of this enzyme in an individual for an accurate interpretation of biological monitoring of exposure to NMP when relying on 5HNMP and/or 2HMSI determination in urine.
Industrial Toxicology and Occupational Medicine Unit, Catholic University of Louvain, Clos Chapelle-aux-Champs 30-54, 1200, Brussels, Belgium. Danuta.Ligocka@toxi.ucl.ac.be
Danuta
D
Ligocka
Dominique
D
Lison
Vincent
V
Haufroid
D000279:Administration, Cutaneous; D000328:Adult; D000818:Animals; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism* / Q000633:toxicity; D065691:Cytochrome P-450 CYP2E1 Inhibitors; D004050:Ditiocarb / Q000494:pharmacology; D004785:Environmental Pollutants / Q000493:pharmacokinetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D010328:Patch Tests; D011760:Pyrrolidinones / Q000008:administration & dosage / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012869:Skin Absorption / Q000187:drug effects; D013723:Teratogens / Q000493:pharmacokinetics*
D065691:Cytochrome P-450 CYP2E1 Inhibitors; D004785:Environmental Pollutants; D004791:Enzyme Inhibitors; D011760:Pyrrolidinones; D013723:Teratogens; D004050:Ditiocarb; D019392:Cytochrome P-450 CYP2E1; C038678:N-methylpyrrolidone