title: Effect of commonly used organic solvents on the kinetics of cytochrome P450 2B6- and 2C8-dependent activity in human liver microsomes.
abstract: The effect of common organic solvents on the activities of various human cytochromes P450 has been reported. However, very little is known about their influence on CYP2B6 and CYP2C8 enzymes. The purpose of this study was to investigate the effect of solvents on the kinetics of representative CYP2B6 (bupropion hydroxylase) and CYP2C8 (paclitaxel hydroxylase) reactions in human liver microsomes. Methanol, ethanol, dimethyl sulfoxide (DMSO), and acetonitrile were studied at increasing volumes (v/v). Acetonitrile, DMSO, and ethanol were shown to increase the Km and decrease the intrinsic clearance (CLint) of CYP2B6-mediated bupropion hydroxylation in a concentration-dependent manner. These solvents did not noticeably alter the Vmax at concentrations of < or =1% (v/v). Unlike the other solvents studied, the effect of methanol (< or =0.5%, v/v) on CYP2B6 kinetics was negligible. Both DMSO and ethanol increased the Km and decreased the CL(int) of CYP2C8-mediated paclitaxel hydroxylation in a concentration-dependent manner. Acetonitrile had minimal influence on CYP2C8 enzyme kinetics at concentrations of < or =1% (v/v). Methanol decreased the Km of paclitaxel at low concentrations followed by an increase at concentrations of > or =2% (v/v). This differential influence on Km resulted in an increased CLint at low concentrations followed by a decrease at high concentrations. The studied solvents had minimal influence on Vmax of paclitaxel. Collectively, DMSO and ethanol were not suitable for characterizing CYP2B6- and CYP2C8-mediated reactions because they showed concentration-dependent inhibition. Methanol and acetonitrile at concentrations of < or =0.5% and < or =1% (v/v) appeared to be suitable for the measurement of CYP2B6- and CYP2C8-mediated activities, respectively.
authors: Metabolism and Pharmacokinetics, Department of Pharmaceutical Candidate Optimization, Bristol-Myer's Squibb Co., P.O. Box 4000, Princeton, NJ 08543, USA. ragini.vuppugalla@bms.com
authors: Ragini
authors: R
authors: Vuppugalla
authors: Shu-Ying
authors: SY
authors: Chang
authors: Hongjian
authors: H
authors: Zhang
authors: Punit H
authors: PH
authors: Marathe
authors: David A
authors: DA
authors: Rodrigues
mesh_terms: D000097:Acetonitriles / Q000494:pharmacology; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D016642:Bupropion / Q000031:analogs & derivatives / Q000378:metabolism; D065702:Cytochrome P-450 CYP2B6; D065727:Cytochrome P-450 CYP2C8; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D000431:Ethanol / Q000494:pharmacology; D006801:Humans; D006900:Hydroxylation / Q000187:drug effects; D007700:Kinetics; D000432:Methanol / Q000494:pharmacology; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D017239:Paclitaxel / Q000378:metabolism; D012997:Solvents / Q000494:pharmacology*; D043823:Taxoids / Q000378:metabolism
chemical_list: D000097:Acetonitriles; D012997:Solvents; D043823:Taxoids; D016642:Bupropion; C086219:6-hydroxytaxol; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; C585599:CYP2B6 protein, human; C450259:CYP2C8 protein, human; D065702:Cytochrome P-450 CYP2B6; D065727:Cytochrome P-450 CYP2C8; D017239:Paclitaxel; C496215:radafaxine; D000432:Methanol; D004121:Dimethyl Sulfoxide; C032159:acetonitrile