Involvement of CYP2J2 and CYP4F12 in the metabolism of ebastine in human intestinal microsomes.
The purpose of the study was to elucidate human intestinal cytochrome P450 isoform(s) involved in the metabolism of an antihistamine, ebastine, having two major pathways of hydroxylation and N-dealkylation. The ebastine dealkylase in human intestinal microsomes was CYP3A4, based on the inhibition studies with antibodies against CYP1A, CYP2A, CYP2C, CYP2D, CYP2E, and CYP3A isoforms and their selective inhibitors. However, ebastine hydroxylase could not be identified. We then examined the inhibitory effects of anti-CYP4F antibody and 17-octadecynoic acid, an inhibitor of the CYP4 family, on ebastine hydroxylation in intestinal microsomes, since CYP4F was recently found to be the predominant ebastine hydroxylase in monkey intestine; and a novel CYP4F isoform (CYP4F12), also capable of hydroxylating ebastine, was found to exist in human intestine. However, the inhibitory effects were only partial (about 20%) and thus it was thought that, although human CYP4F was involved in ebastine hydroxylation, another predominant enzyme exists. Further screening showed that the hydroxylation was inhibited by arachidonic acid. CYP2J2 was selected as a candidate expressed in the intestine and closely related to arachidonic acid metabolism. The catalytic activity of recombinant CYP2J2 was much higher than that of CYP4F12. Anti-CYP2J antibody inhibited the hydroxylation to about 70% in human intestinal microsomes. These results demonstrate that CYP2J2 is the predominant ebastine hydroxylase in human intestinal microsomes. Thus, the present paper for the first time indicates that, in human intestinal microsomes, both CYP2J and CYP4F subfamilies not only metabolize endogenous substrates but also are involved in the drug metabolism.
Hashizume
Takanori
T
Pharmacokinetics and Physico-Chemical Property Research Laboratories, Dainippon Pharmaceutical Company, Ltd., Osaka, Japan. hashidume@dainippon-pharm.co.jp
Imaoka
Susumu
S
Mise
Masashi
M
Terauchi
Yoshiaki
Y
Fujii
Toshihiko
T
Miyazaki
Hisashi
H
Kamataki
Tetsuya
T
Funae
Yoshihiko
Y
D019138:Antibodies, Blocking; D016718:Arachidonic Acid / Q000494:pharmacology; D001189:Aryl Hydrocarbon Hydroxylases; D002090:Butyrophenones / Q000378:metabolism*; D003001:Cloning, Molecular; D000090762:Cytochrome P-450 CYP2J2; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D003640:Dealkylation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006634:Histamine H1 Antagonists / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000201:enzymology / Q000648:ultrastructure; D007700:Kinetics; D007975:Leukotriene B4 / Q000494:pharmacology; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010105:Oxygenases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010880:Piperidines / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
D019138:Antibodies, Blocking; D002090:Butyrophenones; C000717350:CYP2J2 protein, human; D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D006634:Histamine H1 Antagonists; D010880:Piperidines; D011994:Recombinant Proteins; D007975:Leukotriene B4; D016718:Arachidonic Acid; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D010105:Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C422180:CYP4F12 protein, human; D000090762:Cytochrome P-450 CYP2J2; C058249:ebastine