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Xenobiotica
the fate of foreign compounds in biological systems
Volume 45, 2015 - Issue 11
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General Xenobiochemistry

A comprehensive assay for nine major cytochrome P450 enzymes activities with 16 probe reactions on human liver microsomes by a single LC/MS/MS run to support reliable in vitro inhibitory drug–drug interaction evaluation

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Pages 961-977 | Received 12 Feb 2015, Accepted 30 Mar 2015, Published online: 26 May 2015

References

  • Bapiro TE, Egnell AC, Hasler JA, Masimirembwa CM. (2001). Application of higher throughput screening (HTS) inhibition assays to evaluate the interaction of antiparasitic drugs with cytochrome P450s. Drug Metab Dispos 29:30–5
  • Busby WF Jr, Ackermann JM, Crespi CL. (1999). Effect of methanol, ethanol, dimethyl sulfoxide, and acetonitrile on in vitro activities of cDNA-expressed human cytochromes P-450. Drug Metab Dispos 27:246–9
  • Chauret N, Gauthier A, Nicoll-Griffith DA. (1998). Effect of common organic solvents on in vitro cytochrome P450-mediated metabolic activities in human liver microsomes. Drug Metab Dispos 26:1–4
  • De Bock L, Boussery K, Colin P, et al. (2012). Development and validation of a fast and sensitive UPLC–MS/MS method for the quantification of six probe metabolites for the in vitro determination of cytochrome P450 activity. Talanta 89:209–16
  • Dierks EA, Stams KR, Lim HK, et al. (2001). A method for the simultaneous evaluation of the activities of seven major human drug-metabolizing cytochrome P450s using an in vitro cocktail of probe substrates and fast gradient liquid chromatography tandem mass spectrometry. Drug Metab Dispos 29:23–9
  • Dinger J, Meyer MR, Maurer HH. (2014). Development and validation of a liquid-chromatography high-resolution tandem mass spectrometry approach for quantification of nine cytochrome P450 (CYP) model substrate metabolites in an in vitro CYP inhibition cocktail. Anal Bioanal Chem 406:4453–64
  • Dixit V, Hariparsad N, Desai P, Unadkat JD. (2007). In vitro LC–MS cocktail assays to simultaneously determine human cytochrome P450 activities. Biopharm Drug Dispos 28:257–62
  • Donato MT, Jimenez N, Castell JV, Gomez-Lechon MJ. (2004). Fluorescence-based assays for screening nine cytochrome P450 (P450) activities in intact cells expressing individual human P450 enzymes. Drug Metab Dispos 32:699–706
  • Eagling VA, Tjia JF, Back DJ. (1998). Differential selectivity of cytochrome P450 inhibitors against probe substrates in human and rat liver microsomes. Br J Clin Pharmacol 45:107–14
  • Echizen H, Tanizaki M, Tatsuno J, et al. (2000). Identification of CYP3A4 as the enzyme involved in the mono-N-dealkylation of disopyramide enantiomers in humans. Drug Metab Dispos 28:937–44
  • Foti RS, Wahlstrom JL. (2008). CYP2C19 inhibition: the impact of substrate probe selection on in vitro inhibition profiles. Drug Metab Dispos 36:523–8
  • Gonzalez-Perez V, Connolly EA, Bridges AS, et al. (2012). Impact of organic solvents on cytochrome P450 probe reactions: filling the gap with (S)-Warfarin and midazolam hydroxylation. Drug Metab Dispos 40:2136–42
  • Guengerich FP. (2007). Mechanisms of cytochrome P450 substrate oxidation: MiniReview. J Biochem Mol Toxicol 21:163–8
  • Hartman JH, Knott K, Miller GP. (2014). CYP2E1 hydroxylation of aniline involves negative cooperativity. Biochem Pharmacol 87:523–33
  • He F, Bi HC, Xie ZY, et al. (2007). Rapid determination of six metabolites from multiple cytochrome P450 probe substrates in human liver microsome by liquid chromatography/mass spectrometry: application to high-throughput inhibition screening of terpenoids. Rapid Commun Mass Spectrom 21:635–43
  • Hesse LM, Venkatakrishnan K, Court MH, et al. (2000). CYP2B6 mediates the in vitro hydroxylation of bupropion: potential drug interactions with other antidepressants. Drug Metab Dispos 28:1176–83
  • Higashi E, Nakajima M, Katoh M, et al. (2007). Inhibitory effects of neurotransmitters and steroids on human CYP2A6. Drug Metab Dispos 35:508–14
  • Huang SM, Strong JM, Zhang L, et al. (2008). New era in drug interaction evaluation: US Food and Drug Administration update on CYP enzymes, transporters, and the guidance process. J Clin Pharm 48:662–70
  • Huang SM, Temple R, Throckmorton DC, Lesko LJ. (2007). Drug interaction studies: study design, data analysis, and implications for dosing and labeling. Clin Pharm Ther 81:298–304
  • Hummel MA, Gannett PM, Aguilar JS, Tracy TS. (2004). Effector-mediated alteration of substrate orientation in cytochrome P450 2C9. Biochemistry 43:7207–14
  • Hummel MA, Gannett PM, Aguilar J, Tracy TS. (2008). Substrate proton to heme distances in CYP2C9 allelic variants and alterations by the heterotropic activator, dapsone. Arch Biochem Biophys 475:175–83
  • Hutzler JM, Balogh LM, Zientek M, et al. (2009). Mechanism-based inactivation of cytochrome P450 2C9 by tienilic acid and (+/−)-suprofen: a comparison of kinetics and probe substrate selection. Drug Metab Dispos 37:59–65
  • Hutzler JM, Frye RF, Korzekwa KR, et al. (2001a). Minimal in vivo activation of CYP2C9-mediated flurbiprofen metabolism by dapsone. Eur J Pharm Sci 14:47–52
  • Hutzler JM, Hauer MJ, Tracy TS. (2001b). Dapsone activation of CYP2C9-mediated metabolism: evidence for activation of multiple substrates and a two-site model. Drug Metab Dispos 29:1029–34
  • Hutzler JM, Kolwankar D, Hummel MA, Tracy TS. (2002). Activation of CYP2C9-mediated metabolism by a series of dapsone analogs: kinetics and structural requirements. Drug Metab Dispos 30:1194–200
  • Hutzler JM, Tracy TS. (2002). Atypical kinetic profiles in drug metabolism reactions. Drug Metab Dispos 30:355–62
  • Ibeanu GC, Ghanayem BI, Linko P, et al. (1996). Identification of residues 99, 220, and 221 of human cytochrome P450 2C19 as key determinants of omeprazole activity. J Biol Chem 271:12496–501
  • Johnson WW. (2008). Cytochrome P450 inactivation by pharmaceuticals and phytochemicals: therapeutic relevance. Drug Metab Rev 40:101–47
  • Kawalek JC, Andrews AW. (1980). The effect of solvents on drug metabolism in vitro. Drug Metab Disposition 8:380–4
  • Kenworthy KE, Bloomer JC, Clarke SE, Houston JB. (1999). CYP3A4 drug interactions: correlation of 10 in vitro probe substrates. Br J Clin Pharmacol 48:716–27
  • Kim MJ, Kim H, Cha IJ, et al. (2005). High-throughput screening of inhibitory potential of nine cytochrome P450 enzymes in vitro using liquid chromatography/tandem mass spectrometry. Rapid commun Mass Spectrom 19:2651–8
  • Ko JW, Sukhova N, Thacker D, et al. (1997). Evaluation of omeprazole and lansoprazole as inhibitors of cytochrome P450 isoforms. Drug Metab Dispos 25:853–62
  • Korzekwa KR, Krishnamachary N, Shou M, et al. (1998). Evaluation of atypical cytochrome P450 kinetics with two-substrate models: evidence that multiple substrates can simultaneously bind to cytochrome P450 active sites. Biochemistry 37:4137–47
  • Kozakai K, Yamada Y, Oshikata M, et al. (2012). Reliable high-throughput method for inhibition assay of 8 cytochrome P450 isoforms using cocktail of probe substrates and stable isotope-labeled internal standards. Drug Metab Pharmacokinet 27:520–9
  • Kumar V, Wahlstrom JL, Rock DA, et al. (2006). CYP2C9 inhibition: impact of probe selection and pharmacogenetics on in vitro inhibition profiles. Drug Metab Dispos 34:1966–75
  • Lee KS, Kim SK. (2013). Direct and metabolism-dependent cytochrome P450 inhibition assays for evaluating drug–drug interactions. J Appl Toxicol 33:00–8
  • Li XQ, Bjorkman A, Andersson TB, et al. (2002). Amodiaquine clearance and its metabolism to N-desethylamodiaquine is mediated by CYP2C8: a new high affinity and turnover enzyme-specific probe substrate. J Pharm Exp Ther 300:399–407
  • Li T, Li N, Guo Q, et al. (2011). Inhibitory effects of wogonin on catalytic activity of cytochrome P450 enzyme in human liver microsomes. Eur J Drug Metab Pharmacokinet 36:249–56
  • Liu KH, Kim MJ, Shon JH, et al. (2005). Stereoselective inhibition of cytochrome P450 forms by lansoprazole and omeprazole in vitro. Xenobiotica 35:27–38
  • Ludden LK, Ludden TM, Collins JM, et al. (1997). Effect of albumin on the estimation, in vitro, of phenytoin Vmax and Km values: implications for clinical correlation. J Pharm Exp Ther 282:391–6
  • Manevski N, Moreolo PS, Yli-Kauhaluoma J, Finel M. (2011). Bovine serum albumin decreases Km values of human UDP-glucuronosyltransferases 1A9 and 2B7 and increases Vmax values of UGT1A9. Drug Metab Dispos 39:2117–29
  • Masimirembwa CM, Otter C, Berg M, et al. (1999). Heterologous expression and kinetic characterization of human cytochromes P-450: validation of a pharmaceutical tool for drug metabolism research. Drug Metab Dispos 27:1117–22
  • Mo SL, Zhou ZW, Yang LP, et al. (2009). New insights into the structural features and functional relevance of human cytochrome P450 2C9. Part I. Curr Drug Metab 10:1075–126
  • Nebert DW, Russell DW. (2002). Clinical importance of the cytochromes P450. Lancet 360:1155–62
  • Nishiya Y, Nakamura K, Okudaira N, et al. (2010). Effects of organic solvents on the time-dependent inhibition of CYP3A4 by diazepam. Xenobiotica 40:1–8
  • O'Donnell CJ, Grime K, Courtney P, et al. (2007). The development of a cocktail CYP2B6, CYP2C8, and CYP3A5 inhibition assay and a preliminary assessment of utility in a drug discovery setting. Drug Metab Dispos 35:381–5
  • Otten JN, Hingorani GP, Hartley DP, et al. (2011). An in vitro, high throughput, seven CYP cocktail inhibition assay for the evaluation of new chemical entities using LC–MS/MS. Drug Metab Lett 5:17–24
  • Patki KC, Von Moltke LL, Greenblatt DJ. (2003). In vitro metabolism of midazolam, triazolam, nifedipine, and testosterone by human liver microsomes and recombinant cytochromes p450: role of cyp3a4 and cyp3a5. Drug Metab Dispos 31:938–44
  • Pelkonen O, Turpeinen M, Uusitalo J, et al. (2005). Prediction of drug metabolism and interactions on the basis of in vitro investigations. Basic Clin Pharmacol Toxicol 96:167–75
  • Qin CZ, Ren X, Tan ZR, et al. (2014). A high-throughput inhibition screening of major human cytochrome P450 enzymes using an in vitro cocktail and liquid chromatography–tandem mass spectrometry. Biomed chromatogr 28:197–203
  • Ren S, Zeng J, Mei Y, et al. (2013). Discovery and characterization of novel, potent, and selective cytochrome P450 2J2 inhibitors. Drug Metab Dispos 41:60–71
  • Rowland A, Elliot DJ, Knights KM, et al. (2008). The “albumin effect” and in vitro–in vivo extrapolation: sequestration of long-chain unsaturated fatty acids enhances phenytoin hydroxylation by human liver microsomal and recombinant cytochrome P450 2C9. Drug Metab Dispos 36:870–7
  • Rowland A, Gaganis P, Elliot DJ, et al. (2007). Binding of inhibitory fatty acids is responsible for the enhancement of UDP-glucuronosyltransferase 2B7 activity by albumin: implications for in vitro–in vivo extrapolation. J Pharmacol Exp Ther 321:137–47
  • Schrag ML, Wienkers LC. (2001). Covalent alteration of the CYP3A4 active site: evidence for multiple substrate binding domains. Arch Biochem Biophys 391:49–55
  • Sevior DK, Hokkanen J, Tolonen A, et al. (2010). Rapid screening of commercially available herbal products for the inhibition of major human hepatic cytochrome P450 enzymes using the N-in-one cocktail. Xenobiotica 40:245–54
  • Shou M, Mei Q, Ettore MW Jr, et al. (1999). Sigmoidal kinetic model for two co-operative substrate-binding sites in a cytochrome P450 3A4 active site: an example of the metabolism of diazepam and its derivatives. Biochem J 340:845–53
  • Stephens ES, Walsh AA, Scott EE. (2012). Evaluation of inhibition selectivity for human cytochrome P450 2A enzymes. Drug Metab Dispos 40:1797–802
  • Tang C, Lin Y, Rodrigues AD, Lin JH. (2002). Effect of albumin on phenytoin and tolbutamide metabolism in human liver microsomes: an impact more than protein binding. Drug Metab Dispos 30:648–54
  • Tang C, Shou M, Rodrigues AD. (2000). Substrate-dependent effect of acetonitrile on human liver microsomal cytochrome P450 2C9 (CYP2C9) activity. Drug Metab Dispos 28:567–72
  • Testino SA Jr, Patonay G. (2003). High-throughput inhibition screening of major human cytochrome P450 enzymes using an in vitro cocktail and liquid chromatography–tandem mass spectrometry. J Pharm Biomed Anal 30:1459–67
  • Tolonen A, Petsalo A, Turpeinen M, et al. (2007). In vitro interaction cocktail assay for nine major cytochrome P450 enzymes with 13 probe reactions and a single LC/MSMS run: analytical validation and testing with monoclonal anti-CYP antibodies. J Mass Spectrom 42:960–6
  • Tsao CC, Wester MR, Ghanayem B, et al. (2001). Identification of human CYP2C19 residues that confer S-mephenytoin 4′-hydroxylation activity to CYP2C9. Biochemistry 40:1937–44
  • Turpeinen M, Uusitalo J, Jalonen J, Pelkonen O. (2005). Multiple P450 substrates in a single run: rapid and comprehensive in vitro interaction assay. Eur J Pharm Sci 24:123–32
  • VandenBrink BM, Foti RS, Rock DA, et al. (2012). Prediction of CYP2D6 drug interactions from in vitro data: evidence for substrate-dependent inhibition. Drug Metab Dispos 40:47–53
  • Vuppugalla R, Chang SY, Zhang H, et al. (2007). Effect of commonly used organic solvents on the kinetics of cytochrome P450 2B6- and 2C8-dependent activity in human liver microsomes. Drug Metab Dispos 35:1990–5
  • Walsky RL, Obach RS. (2004). Validated assays for human cytochrome P450 activities. Drug Metab Dispos 32:647–60
  • Wang Y, Unadkat JD. (1999). Enzymes in addition to CYP3A4 and 3A5 mediate N-demethylation of dextromethorphan in human liver microsomes. Biopharm Drug Dispos 20:341–6
  • Wang L, Zhang D, Raghavan N, et al. (2010). In vitro assessment of metabolic drug–drug interaction potential of apixaban through cytochrome P450 phenotyping, inhibition, and induction studies. Drug Metab Dispos 38:448–58
  • Wattanachai N, Polasek TM, Heath TM, et al. (2011). In vitro–in vivo extrapolation of CYP2C8-catalyzed paclitaxel 6alpha-hydroxylation: effects of albumin on in vitro kinetic parameters and assessment of interindividual variability in predicted clearance. Eur J Clin Pharmacol 67:815–24
  • Wattanachai N, Tassaneeyakul W, Rowland A, et al. (2012). Effect of albumin on human liver microsomal and recombinant CYP1A2 activities: impact on in vitro–in vivo extrapolation of drug clearance. Drug Metab Dispos 40:982–9
  • Williams PA, Cosme J, Ward A, et al. (2003). Crystal structure of human cytochrome P450 2C9 with bound warfarin. Nature 424:464–8
  • Yao M, Zhu M, Sinz MW, et al. (2007). Development and full validation of six inhibition assays for five major cytochrome P450 enzymes in human liver microsomes using an automated 96-well microplate incubation format and LC–MS/MS analysis. J Pharm Biomed Anal 44:211–23
  • Yoda N, Emoto C, Date S, et al. (2012). Characterization of intestinal and hepatic P450 enzymes in cynomolgus monkeys with typical substrates and inhibitors for human P450 enzymes. Xenobiotica 42:719–30
  • Yuan R, Madani S, Wei XX, et al. (2002). Evaluation of cytochrome P450 probe substrates commonly used by the pharmaceutical industry to study in vitro drug interactions. Drug Metab Dispos 30:1311–19
  • Zambon S, Fontana S, Kajbaf M. (2010). Evaluation of cytochrome P450 inhibition assays using human liver microsomes by a cassette analysis /LC–MS/MS. Drug Metab Lett 4:120–8
  • Zhang T, Zhu Y, Gunaratna C. (2002). Rapid and quantitative determination of metabolites from multiple cytochrome P450 probe substrates by gradient liquid chromatography–electrospray ionization-ion trap mass spectrometry. J Chromatogr B 780:371–9
  • Zhao P, Kunze KL, Lee CA. (2005). Evaluation of time-dependent inactivation of CYP3A in cryopreserved human hepatocytes. Drug Metab Dispos 33:853–61
  • Zhou S, Yung CS, Cher GB, et al. (2005). Mechanism-based inhibition of cytochrome P450 3A4 by therapeutic drugs. Clin Pharmacokinet 44:279–304
  • Zhou SF, Zhou ZW, Yang LP, Cai JP. (2009). Substrates, inducers, inhibitors and structure-activity relationships of human Cytochrome P450 2C9 and implications in drug development. Curr Med Chem 16:3480–675
  • Zientek M, Miller H, Smith D, et al. (2008). Development of an in vitro drug–drug interaction assay to simultaneously monitor five cytochrome P450 isoforms and performance assessment using drug library compounds. J Pharmacol Toxicol Methods 58:206–14

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