Intestinal and hepatic CYP3A4 catalyze hydroxylation of 1alpha,25-dihydroxyvitamin D(3): implications for drug-induced osteomalacia.
The decline in bone mineral density that occurs after long-term treatment with some antiepileptic drugs is thought to be mediated by increased vitamin D(3) metabolism. In this study, we show that the inducible enzyme CYP3A4 is a major source of oxidative metabolism of 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] in human liver and small intestine and could contribute to this adverse effect. Heterologously-expressed CYP3A4 catalyzed the 23- and 24-hydroxylation of 1,25(OH)(2)D(3). No human microsomal cytochrome P450 enzyme tested, other than CYP3A5, supported these reactions. CYP3A4 exhibited opposite product stereochemical preference compared with that of CYP24A1, a known 1,25(OH)(2)D(3) hydroxylase. The three major metabolites generated by CYP3A4 were 1,23R,25(OH)(3)D(3), 1,24S,25(OH)(3)D(3), and 1,23S,25(OH)(3)D(3). Although the metabolic clearance of CYP3A4 was less than that of CYP24A1, comparison of metabolite profiles and experiments using CYP3A-specific inhibitors indicated that CYP3A4 was the dominant source of 1,25(OH)(2)D(3) 23- and 24-hydroxylase activity in both human small intestine and liver. Consistent with this observation, analysis of mRNA isolated from human intestine and liver (including samples from donors treated with phenytoin) revealed a general absence of CYP24A1 mRNA. In addition, expression of CYP3A4 mRNA in a panel of duodenal samples was significantly correlated with the mRNA level of a known vitamin D receptor gene target, calbindin-D9K. These and other data suggest that induction of CYP3A4-dependent 1,25(OH)(2)D(3) metabolism by antiepileptic drugs and other PXR ligands may diminish intestinal effects of the hormone and contribute to osteomalacia.
Xu
Yang
Y
Department of Pharmaceutics, Box 357610, University of Washington, Seattle, WA 98195-7610, USA.
Hashizume
Takanori
T
Shuhart
Margaret C
MC
Davis
Connie L
CL
Nelson
Wendel L
WL
Sakaki
Toshiyuki
T
Kalhorn
Thomas F
TF
Watkins
Paul B
PB
Schuetz
Erin G
EG
Thummel
Kenneth E
KE
D001483:Base Sequence; D018938:Caco-2 Cells; D064026:Calbindins; D002117:Calcitriol / Q000378:metabolism*; D002384:Catalysis; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D017931:DNA Primers; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006900:Hydroxylation; D007421:Intestine, Small / Q000201:enzymology*; D007654:Ketoconazole / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008874:Midazolam / Q000378:metabolism; D010018:Osteomalacia / Q000188:drug therapy*; D011994:Recombinant Proteins / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D014217:Troleandomycin / Q000494:pharmacology
D064026:Calbindins; D017931:DNA Primers; D011994:Recombinant Proteins; D064030:S100 Calcium Binding Protein G; C576506:S100G protein, human; D003577:Cytochrome P-450 Enzyme System; D014217:Troleandomycin; C104464:CYP3A protein, human; C510164:CYP3A5 protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D002117:Calcitriol; D008874:Midazolam; D007654:Ketoconazole