Studies on induction of lamotrigine metabolism in transgenic UGT1 mice.
A transgenic 'knock-in' mouse model expressing a human UGT1 locus (Tg-UGT1) was recently developed and validated. Although these animals express mouse UGT1A proteins, UGT1A4 is a pseudo-gene in mice. Therefore, Tg-UGT1 mice serve as a 'humanized' UGT1A4 animal model. Lamotrigine (LTG) is primarily metabolized to its N-glucuronide (LTGG) by hUGT1A4. This investigation aimed at examining the impact of pregnane X receptor (PXR), constitutive androstane receptor (CAR) and peroxisome proliferator-activated receptor (PPAR) activators on LTG glucuronidation in vivo and in vitro. Tg-UGT1 mice were administered the inducers phenobarbital (CAR), pregnenolone-16alpha-carbonitrile (PXR), WY-14643 (PPAR-alpha), ciglitazone (PPAR-gamma), or L-165041 (PPAR-beta), once daily for 3 or 4 days. Thereafter, LTG was administered orally and blood samples were collected over 24 h. LTG was measured in blood and formation of LTGG was measured in pooled microsomes made from the livers of treated animals. A three-fold increase in in vivo LTG clearance was seen after phenobarbital administration. In microsomes prepared from phenobarbital-treated Tg-UGT1 animals, 13-fold higher CL(int) (Vmax/K(m)) value was observed as compared with the untreated transgenic mice. A trend toward induction of catalytic activity in vitro and in vivo was also observed following pregnenolone-16alpha-carbonitrile and WY-14643 treatment. This study demonstrates the successful application of Tg-UGT1 mice as a novel tool to study the impact of induction and regulation on metabolism of UGT1A4 substrates.
Argikar
U A
UA
Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota, Minneapolis, MN 55455, USA.
Senekeo-Effenberger
K
K
Larson
E E
EE
Tukey
R H
RH
Remmel
R P
RP
D000818:Animals; D000927:Anticonvulsants / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid; D004790:Enzyme Induction / Q000187:drug effects / Q000502:physiology; D005260:Female; D018757:GABA Modulators / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D000077213:Lamotrigine; D008297:Male; D008657:Metabolic Clearance Rate / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D047492:Peroxisome Proliferator-Activated Receptors / Q000494:pharmacology; D020025:Peroxisome Proliferators / Q000494:pharmacology; D010634:Phenobarbital / Q000494:pharmacology; D011285:Pregnenolone Carbonitrile / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D014227:Triazines / Q000493:pharmacokinetics*
D000927:Anticonvulsants; D018757:GABA Modulators; D047492:Peroxisome Proliferator-Activated Receptors; D020025:Peroxisome Proliferators; D011743:Pyrimidines; D014227:Triazines; C019787:bilirubin glucuronoside glucuronosyltransferase; D011285:Pregnenolone Carbonitrile; C006253:pirinixic acid; D014453:Glucuronosyltransferase; D000077213:Lamotrigine; D010634:Phenobarbital