title: Inhibitory effects and oxidation of 6-methylcoumarin, 7-methylcoumarin and 7-formylcoumarin via human CYP2A6 and its mouse and pig orthologous enzymes.
abstract: 1. Information about the metabolism of compounds is essential in drug discovery and development, risk assessment of chemicals and further development of predictive methods. 2. In vitro and in silico methods were applied to evaluate the metabolic and inhibitory properties of 6-methylcoumarin, 7-methylcoumarin and 7-formylcoumarin with human CYP2A6, mouse CYP2A5 and pig CYP2A19. 3. 6-Methylcoumarin was oxidized to fluorescent 7-hydroxy-6-methylcoumarin by CYP2A6 (Km: 0.64-0.91 µM; Vmax: 0.81-0.89 min(-1)) and by CYP2A5 and CYP2A19. The reaction was almost completely inhibited at 10 µM 7-methylcoumarin in liver microsomes of human and mouse, but in pig only 40% inhibition was obtained with the anti-CYP2A5 antibody or with methoxsalen and pilocarpine. 7-Methylcoumarin was a mechanism-based inhibitor for CYP2A6, but not for the mouse and pig enzymes. 7-Formylcoumarin was a mechanism-based inhibitor for CYP2As of all species. 4. Docking and molecular dynamics simulations of 6-methylcoumarin and 7-methylcoumarin in the active sites of CYP2A6 and CYP2A5 demonstrated a favorable orientation of the 7-position of 6-methylcoumarin towards the heme moiety. Several orientations of 7-methylcoumarin were possible in CYP2A6 and CYP2A5. 5. These results indicate that the active site of CYP2A6 has unique interaction properties for ligands and differs in this respect from CYP2A5 and CYP2A19.
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.
authors.forename: Risto O
authors.initials: RO
authors.lastname: Juvonen
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.|b Department of Biological and Environmental Science & Nanoscience Center , University of Jyvaskyla , Jyvaskyla , Finland.
authors.forename: Mira
authors.initials: M
authors.lastname: Kuusisto
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.
authors.forename: Carolin
authors.initials: C
authors.lastname: Fohrgrup
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.
authors.forename: Mari H
authors.initials: MH
authors.lastname: Pitkänen
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.
authors.forename: Tapio J
authors.initials: TJ
authors.lastname: Nevalainen
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.
authors.forename: Seppo
authors.initials: S
authors.lastname: Auriola
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.
authors.forename: Hannu
authors.initials: H
authors.lastname: Raunio
authors.affiliation: a Faculty of Health Sciences , School of Pharmacy, University of Eastern Finland , Kuopio , Finland and.
authors.forename: Markku
authors.initials: M
authors.lastname: Pasanen
authors.affiliation: b Department of Biological and Environmental Science & Nanoscience Center , University of Jyvaskyla , Jyvaskyla , Finland.
authors.forename: Olli T
authors.initials: OT
authors.lastname: Pentikäinen
mesh_terms: D000818:Animals; D003374:Coumarins / Q000494:pharmacology*; D065637:Cytochrome P-450 CYP2A6 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D065607:Cytochrome P-450 Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D006900:Hydroxylation; D020128:Inhibitory Concentration 50; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D010084:Oxidation-Reduction; D034421:Sus scrofa; D013997:Time Factors
chemical_list: D003374:Coumarins; D065607:Cytochrome P-450 Enzyme Inhibitors; C585710:CYP2A6 protein, human; D065637:Cytochrome P-450 CYP2A6; C018096:6-methylcoumarin; C580890:7-methylcoumarin
keywords: Coumarin derivative; cytochrome P450; enzyme assay; fluorescence; metabolism