User menu

Accès à distance ? S'identifier sur le proxy UCLouvain

Contribution of CYP2E1 to N-methyl-2-pyrrolidone metabolism.

Primary tabs

Ligocka, Danuta [UCL] Lison, Dominique [UCL] Haufroid, Vincent [UCL]

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.

  1. Åkesson B, Jönsson BA (1997) Major metabolic pathway for N-methyl-2-pyrrolidone in humans. Drug Metab Dispos 25:267–269
  2. Åkesson B, Jönsson BA (2000a) Dermal absorption study of N-methyl-2-pyrrolidone in male and female volunteers. Scientific Programme and Abstracts of the 26th International Congress on Occupational Health, Singapore, p 312
  3. Åkesson Bengt, Jönsson Bo AG, Biological monitoring of N-methyl-2-pyrrolidone using 5-hydroxy-N-methyl-2-pyrrolidone in plasma and urine as the biomarker, 10.5271/sjweh.534
  4. Akesson B, Paulsson K, Experimental exposure of male volunteers to N-methyl-2-pyrrolidone (NMP): acute effects and pharmacokinetics of NMP in plasma and urine., 10.1136/oem.54.4.236
  5. Beaulieu Harry J., Schmerber Kenneth R., M-Pyrol™(NMP) Use in the Microelectronics Industry, 10.1080/1047322x.1991.10387980
  6. BECCI P, KNICKERBOCKER M, REAGAN E, PARENT R, BURNETTE L, Teratogenicity study of N-methylpyrrolidone after dermal application to Sprague-Dawley rats, 10.1016/s0272-0590(82)80117-6
  7. Chung HC, Sung SH, Kim JS, Kim YC, Kim SG (2001) Lack of cytochrome P450 2E1 (CYP2E1) induction in the rat liver by starvation without coprophagy. Drug Metab Dispos 29:213–216
  8. Frye Reginald F., Branch Robert A., Effect of chronic disulfiram administration on the activities of CYP1A2, CYP2C19, CYP2D6, CYP2E1, and N-acetyltransferase in healthy human subjects : Effect of chronic disulfiram on drug metabolizing enzymes, 10.1046/j.1365-2125.2002.01522.x
  9. Haufroid V, Toubeau F, Clippe A, Buysschaert M, Gala JL, Lison D (2001) Real-time quantification of cytochrome P4502E1 mRNA in human peripheral blood lymphocytes by reverse transcription-PCR: method and practical application. Clin Chem 47:1126–1129
  10. HSDB (2002) Hazardous Substances Data Bank. 1-Methyl-2-pyrrolidinone. US National Library of Medicine, Bethesda MD. Available online via NIH TOXNET http://toxnet.nlm.nih.gov/cgi-bin/sis/search/
  11. Jungbauer F. H. W., Coenraads P. J., Kardaun S. H., Toxic hygroscopic contact reaction to N-methyl-2-pyrrolidone, 10.1034/j.1600-0536.2001.450514.x
  12. Kim RB, Yamazaki H, Chiba D, Oshea D, Mimura M, Guengerich FP, Ishizaki T, Shimada T, Wilkinson GR (1996) In vitro and in vivo characterization of CYP2E1 activity in Japanese and Caucasians. J Pharmacol Exp Ther 279: 4–11
  13. LEE K, Toxicity of N-methyl-2-pyrrolidone (NMP): Teratogenic, subchronic, and two-year inhalation studies, 10.1016/0272-0590(87)90045-5
  14. Leira Hakos L., Tiltnse Arne, Svendsen Kristin, Vetlesen Lars, Irritant cutaneous reactions to N-methyl-2-pyrrolidone (NMP), 10.1111/j.1600-0536.1992.tb05243.x
  15. Lieber, CS (1997) Cytochrome P-450: its physiological and pathological role. Physiol Rev 77:517–544
  16. Ligocka D., Lison D., Haufroid V., Quantitative determination of 5-hydroxy-N-methylpyrrolidone in urine for biological monitoring of N-methylpyrrolidone exposure, 10.1016/s0378-4347(01)00441-8
  17. Lucas Danièle, Berthou François, Girre Catherine, Poitrenaud Frédérique, Ménez Jean-François, High-performance liquid chromatographic determination of chlorzoxazone and 6-hydroxychlorzoxazone in serum: a tool for indirect evaluation of cytochrome P4502E1 activity in humans, 10.1016/0378-4347(93)80252-y
  18. Lucas Daniele, Ferrara Roberta, Gonzalez Eva, Bodenez Pierre, Albores Arnulfo, Manno Maurizio, Berthou Francois, Chlorzoxazone, a selective probe for phenotyping CYP2E1 in humans : , 10.1097/00008571-199906000-00013
  19. Mizuno Dai, Tanaka Einosuke, Tanno Kozo, Misawa Shogo, Chlorzoxazone: a probe drug whose metabolism can be used to monitor toluene exposure in rats, 10.1007/s002040050666
  20. Raucy Judy L., Schultz Eric D., Kearins Margaret C., Arora Sanjeev, Johnston David E., Omdahl John L., Eckmann Lars, Carpenter Susan P., CYP2E1 Expression in Human Lymphocytes From Various Ethnic Populations, 10.1111/j.1530-0277.1999.tb04085.x
  21. Solomon H. M., Burgess B. A., Kennedy G. L., Staples R. E., 1-methyl-2-pyrrolidone (nmp): reproductive and developmental toxicity study by inhalation in the rat, 10.3109/01480549509014324
  22. Wells DA, Digenis GA (1988) Disposition and metabolism of double-labeled [3H and 14C] N-methyl-2- pyrrolidinone in the rat. Drug Metab Dispos 16:243–249
  23. Wells DA, Hawi AA, Digenis GA (1992) Isolation and identification of the major urinary metabolite of N-methylpyrrolidinone in the rat. Drug Metab Dispos 20:124–126
Bibliographic reference Ligocka, Danuta ; Lison, Dominique ; Haufroid, Vincent. Contribution of CYP2E1 to N-methyl-2-pyrrolidone metabolism.. In: Archives of toxicology, Vol. 77, no. 5, p. 261-266 (2003)
Permanent URL http://hdl.handle.net/2078.1/22357