Bioactivation of lamotrigine in vivo in rat and in vitro in human liver microsomes, hepatocytes, and epidermal keratinocytes: characterization of thioether conjugates by liquid chromatography/mass spectrometry and high field nuclear magnetic resonance spectroscopy.
Previous studies suggested that lamotrigene (LTG) underwent bioactivation to a reactive aryl epoxide intermediate in rats. Nevertheless, definitive structures of these thioether conjugates, which are often needed to substantiate the mechanism of bioactivation and identity of reactive intermediate(s), were not fully established. In the present study, GSH, cysteinylglycine, and N-acetyl cysteine conjugates of LTG were isolated from bile of rats orally dosed with LTG (100 mg/kg), and their structures were fully elucidated by LC/MS and NMR. The definitive structural characterization of these metabolites provided evidence for the existence of a reactive aryl epoxide that was trapped as a GSH adduct. In vitro studies using various hepatic cellular and subcellular fractions obtained from human and rat were performed to demonstrate that LTG underwent bioactivation to form a GSH conjugate that was identical to the one initially characterized from in vivo studies. Human P450 2A6 and rat P450 2C11 appeared to be the primary enzymes activating LTG in human and rat liver microsomes, respectively. Interindividual variation in the bioactivation of LTG was demonstrated with 20 individual human liver microsomes. Furthermore, it was shown that human epidermal keratinocytes were capable of forming the same GSH conjugate, suggesting that LTG could be bioactivated in skin cells. The results from these studies suggest that LTG has the potential to undergo hepatic and nonhepatic bioactivation, leading to a reactive aryl epoxide intermediate in human. The bioactivation of LTG in epidermal cells provides a possible explanation for the idiosyncratic cutaneous reactions associated with LTG therapy.
Chen
Hao
H
Department of Pharmacokinetics, Dynamics, and Metabolism, Pfizer Global Research and Development, 2800 Plymouth Road, Ann Arbor, Michigan 48105, USA.
Grover
Scott
S
Yu
Linning
L
Walker
Gregory
G
Mutlib
Abdul
A
D000284:Administration, Oral; D000818:Animals; D000927:Anticonvulsants / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D065637:Cytochrome P-450 CYP2A6; D000072467:Cytochrome P450 Family 2; D005260:Female; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D007700:Kinetics; D000077213:Lamotrigine; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D008968:Molecular Conformation; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization; D039181:Steroid 16-alpha-Hydroxylase / Q000378:metabolism; D013440:Sulfides / Q000737:chemistry; D014227:Triazines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
D000927:Anticonvulsants; D013440:Sulfides; D014227:Triazines; D001189:Aryl Hydrocarbon Hydroxylases; C450279:CYP2C11 protein, rat; D065637:Cytochrome P-450 CYP2A6; D000072467:Cytochrome P450 Family 2; D039181:Steroid 16-alpha-Hydroxylase; D005978:Glutathione; D000077213:Lamotrigine