title: Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin: In silico predictions and experimental validation.
abstract: Deoxypodophyllotoxin is stereoselectively converted into epipodophyllotoxin by recombinant human cytochrome P450 3A4 (CYP3A4). Further kinetic analysis revealed that the Michaelis-Menten K(m) and V(max) for hydroxylation of deoxypodophyllotoxin by CYP3A4 at C7 position were 1.93 microM and 1.48 nmol/min/nmol, respectively. Deoxypodophyllotoxin was subjected to automated docking analysis in order to get better knowledge of the interaction between the CYP3A4 enzyme and the substrate, using the PatchDock algorithm with distance constraints. Automated docking showed that the beta-hydrogen atom at C7 position is in the most appropriate binding orientation at the site of oxidation. The docking results are consistent with the experimental data for the bioconversion of deoxypodophyllotoxin into epipodophyllotoxin by CYP3A4. In addition, the effects of five lignans, deoxypodophyllotoxin, epipodophyllotoxin, podophyllotoxin, demethylenedeoxypodophyllotoxin, and demethylenepodophyllotoxin, on CYP3A4 were compared in order to investigate the influence of the methylenedioxy group on the biotransformation process, to give insight into the mode of metabolization and to explain inhibitory activity of lignans.
authors.affiliation: Department of Pharmaceutical Biology, Graduate School for Drug Exploration (GUIDE), University of Groningen, Antonius Deusinglaan 1, NL-9713 AV Groningen, The Netherlands.
authors.forename: Mattijs K
authors.initials: MK
authors.lastname: Julsing
authors.forename: Nikolay P
authors.initials: NP
authors.lastname: Vasilev
authors.forename: Dina
authors.initials: D
authors.lastname: Schneidman-Duhovny
authors.forename: Remco
authors.initials: R
authors.lastname: Muntendam
authors.forename: Herman J
authors.initials: HJ
authors.lastname: Woerdenbag
authors.forename: Wim J
authors.initials: WJ
authors.lastname: Quax
authors.forename: Haim J
authors.initials: HJ
authors.lastname: Wolfson
authors.forename: Iliana
authors.initials: I
authors.lastname: Ionkova
authors.forename: Oliver
authors.initials: O
authors.lastname: Kayser
mesh_terms: D000465:Algorithms; D002851:Chromatography, High Pressure Liquid; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism*; D004365:Drugs, Chinese Herbal; D006801:Humans; D008958:Models, Molecular; D011034:Podophyllotoxin / Q000031:analogs & derivatives* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D004365:Drugs, Chinese Herbal; D011994:Recombinant Proteins; C014451:deoxypodophyllotoxin; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D011034:Podophyllotoxin