Mechanism-Based Inactivation of Human Cytochrome P450 Enzymes and the Prediction of Drug-Drug Interactions

The ability to use vitro inactivation kinetic parameters in scaling to in vivo drug-drug interactions (DDIs) for mechanism-based inactivators of human cytochrome P450 (P450) enzymes was examined using eight human P450-selective marker activities in pooled human liver microsomes. These data were combined with other parameters (systemic Cmax, estimated hepatic inlet Cmax, fraction unbound, in vivo P450 enzyme degradation rate constants estimated from clinical pharmacokinetic data, and fraction of the affected drug cleared by the inhibited enzyme) to predict increases in exposure to drugs, and the predictions were compared with in vivo DDIs gathered from clinical studies reported in the scientific literature. In general, the use of unbound systemic Cmax as the inactivator concentration in vivo yielded the most accurate predictions of DDI with a mean -fold error of 1.64. Abbreviated in vitro approaches to identifying mechanism-based inactivators were developed. Testing potential inactivators at a single concentration (IC25) in a 30-min preincubation with human liver microsomes in the absence and presence of NADPH followed by assessment of P450 marker activities readily identified those compounds known to be mechanism-based inactivators and represents an approach that can be used with greater throughput. Measurement of decreases in IC50 occurring with a 30-min preincubation with liver microsomes and NADPH was also useful in identifying mechanism-based inactivators, and the IC50 measured after such a preincubation was highly correlated with the kinact/KI ratio measured after a full characterization of inactivation. Overall, these findings support the conclusion that P450 in vitro inactivation data are valuable in predicting clinical DDIs that can occur via this mechanism.

[1]  A. Rostami-Hodjegan,et al.  'In silico' simulations to assess the 'in vivo' consequences of 'in vitro' metabolic drug-drug interactions. , 2004, Drug discovery today. Technologies.

[2]  R. O'Reilly.  Ticrynafen‐racemic warfarin interaction: Hepatotoxic or stereoselective? , 1982, Clinical pharmacology and therapeutics.

[3]  J. Segura,et al.  Accumulation of caffeine in healthy volunteers treated with furafylline. , 1987, British journal of clinical pharmacology.

[4]  D. Greenblatt,et al.  Time course of recovery of cytochrome p450 3A function after single doses of grapefruit juice , 2003, Clinical pharmacology and therapeutics.

[5]  J. Houston,et al.  In vitro cytochrome P450 inhibition data and the prediction of drug‐drug interactions: Qualitative relationships, quantitative predictions, and the rank‐order approach , 2005, Clinical pharmacology and therapeutics.

[6]  O. Pelkonen,et al.  Effect of Clopidogrel and Ticlopidine on Cytochrome P450 2B6 Activity as Measured by Bupropion Hydroxylation , 2005, Clinical pharmacology and therapeutics.

[7]  P. Maurel,et al.  Effect of corticosteroids on the expression of cytochromes P450 and on cyclosporin A oxidase activity in primary cultures of human hepatocytes. , 1992, Molecular pharmacology.

[8]  M. Fromm,et al.  Differential induction of prehepatic and hepatic metabolism of verapamil by rifampin , 1996, Hepatology.

[9]  Yvonne S. Lin,et al.  Differences in the inhibition of cytochromes P450 3A4 and 3A5 by metabolite-inhibitor complex-forming drugs. , 2004, Drug metabolism and disposition: the biological fate of chemicals.

[10]  Hayley S. Brown,et al.  Prediction of in vivo drug-drug interactions from in vitro data: impact of incorporating parallel pathways of drug elimination and inhibitor absorption rate constant. , 2005, British journal of clinical pharmacology.

[11]  R Scott Obach,et al.  SELECTIVE INHIBITION OF HUMAN CYTOCHROME P4502C8 BY MONTELUKAST , 2005, Drug Metabolism and Disposition.

[12]  A. Colli,et al.  Ticlopidine‐theophylline interaction , 1987, Clinical pharmacology and therapeutics.

[13]  M. Eichelbaum,et al.  Potent Mechanism-Based Inhibition of Human CYP2B6 by Clopidogrel and Ticlopidine , 2004, Journal of Pharmacology and Experimental Therapeutics.

[14]  L. Dubé,et al.  Effect of Zileuton on Theophylline Pharmacokinetics , 1995, Clinical pharmacokinetics.

[15]  A. D. Rodrigues,et al.  Mechanism-based inhibition of human liver microsomal cytochrome P450 1A2 by zileuton, a 5-lipoxygenase inhibitor. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[16]  J. Houston,et al.  The Utility of in Vitro Cytochrome P450 Inhibition Data in the Prediction of Drug-Drug Interactions , 2006, Journal of Pharmacology and Experimental Therapeutics.

[17]  S. Risch,et al.  Differential Time Course of Cytochrome P450 2D6 Enzyme Inhibition by Fluoxetine, Sertraline, and Paroxetine in Healthy Volunteers , 2002, Journal of clinical psychopharmacology.

[18]  R. Obach,et al.  Validated assays for human cytochrome P450 activities. , 2004, Drug metabolism and disposition: the biological fate of chemicals.

[19]  R. Obach,et al.  IN VITRO-IN VIVO EXTRAPOLATION OF CYP2D6 INACTIVATION BY PAROXETINE: PREDICTION OF NONSTATIONARY PHARMACOKINETICS AND DRUG INTERACTION MAGNITUDE , 2005, Drug Metabolism and Disposition.

[20]  S D Hall,et al.  An in vitro model for predicting in vivo inhibition of cytochrome P450 3A4 by metabolic intermediate complex formation. , 2000, Drug metabolism and disposition: the biological fate of chemicals.

[21]  S. Krähenbühl,et al.  Biotransformation of caffeine by cDNA-expressed human cytochromes P-450 , 2004, European Journal of Clinical Pharmacology.

[22]  G. Tucker,et al.  MECHANISM-BASED INACTIVATION OF CYP2D6 BY METHYLENEDIOXYMETHAMPHETAMINE , 2004, Drug Metabolism and Disposition.

[23]  P. Neuvonen,et al.  Plasma buspirone concentrations are greatly increased by erythromycin and itraconazole , 1997, Clinical pharmacology and therapeutics.

[24]  A. D. Rodrigues,et al.  Drug-drug interactions , 2001, Atkinson's Principles of Clinical Pharmacology.

[25]  L. Sayre,et al.  Mechanism-based inactivation of cytochromes P450 2B1 and P450 2B6 by 2-phenyl-2-(1-piperidinyl)propane. , 2000, Drug metabolism and disposition: the biological fate of chemicals.

[26]  Kiyomi Ito,et al.  Prediction of the in vivo interaction between midazolam and macrolides based on in vitro studies using human liver microsomes. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[27]  Y. Sawada,et al.  Pharmacokinetic analysis of felodipine-grapefruit juice interaction based on an irreversible enzyme inhibition model. , 2000, British journal of clinical pharmacology.

[28]  J. Backman,et al.  Dose of midazolam should be reduced during diltiazem and verapamil treatments. , 1994, British journal of clinical pharmacology.

[29]  J. Miners,et al.  Mechanism-Based Inactivation of Human Cytochrome P4502C8 by Drugs in Vitro , 2004, Journal of Pharmacology and Experimental Therapeutics.

[30]  G. Tucker,et al.  MECHANISM-BASED INACTIVATION OF CYP2D6 BY METHYLENEDIOXYMETHAMPHETAMINE , 2004, Drug Metabolism and Disposition.

[31]  C. Ernest,et al.  Mechanism-Based Inactivation of CYP3A by HIV Protease Inhibitors , 2005, Journal of Pharmacology and Experimental Therapeutics.

[32]  K Rowland-Yeo,et al.  A critical evaluation of the experimental design of studies of mechanism based enzyme inhibition, with implications for in vitro-in vivo extrapolation. , 2006, Current drug metabolism.

[33]  Aleksandra Galetin,et al.  PREDICTION OF TIME-DEPENDENT CYP3A4 DRUG-DRUG INTERACTIONS: IMPACT OF ENZYME DEGRADATION, PARALLEL ELIMINATION PATHWAYS, AND INTESTINAL INHIBITION , 2006, Drug Metabolism and Disposition.

[34]  Y. Sugiyama,et al.  Prediction of In Vivo Interaction Between Triazolam and Erythromycin Based on In Vitro Studies Using Human Liver Microsomes and Recombinant Human CYP3A4 , 2000, Pharmaceutical Research.

[35]  D. Greenblatt,et al.  Apparent mechanism-based inhibition of human CYP2D6 in vitro by paroxetine: comparison with fluoxetine and quinidine. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[36]  U. Fuhr,et al.  Time response of cytochrome P450 1A2 activity on cessation of heavy smoking , 2004, Clinical pharmacology and therapeutics.

[37]  T. Kumai,et al.  Ticlopidine decreases the in vivo activity of CYP2C19 as measured by omeprazole metabolism. , 1999, British journal of clinical pharmacology.

[38]  A. Schwarz,et al.  Pharmacokinetics of meloxicam in patients with end-stage renal failure on haemodialysis: a comparison with healthy volunteers , 1996, European Journal of Clinical Pharmacology.

[39]  M. Jamei,et al.  Kinetic values for mechanism-based enzyme inhibition: assessing the bias introduced by the conventional experimental protocol. , 2005, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[40]  D. Greenblatt,et al.  CYP2B6 mediates the in vitro hydroxylation of bupropion: potential drug interactions with other antidepressants. , 2000, Drug metabolism and disposition: the biological fate of chemicals.

[41]  P. Neuvonen,et al.  Effects of verapamil and diltiazem on the pharmacokinetics and pharmacodynamics of buspirone , 1998, Clinical pharmacology and therapeutics.

[42]  S. Hall,et al.  Prediction of cytochrome P450 3A inhibition by verapamil enantiomers and their metabolites. , 2004, Drug metabolism and disposition: the biological fate of chemicals.

[43]  Kiyomi Ito,et al.  IMPACT OF PARALLEL PATHWAYS OF DRUG ELIMINATION AND MULTIPLE CYTOCHROME P450 INVOLVEMENT ON DRUG-DRUG INTERACTIONS: CYP2D6 PARADIGM , 2005, Drug Metabolism and Disposition.

[44]  D. Greenblatt,et al.  Desipramine pharmacokinetics when coadministered with paroxetine or sertraline in extensive metabolizers. , 1997, Journal of clinical psychopharmacology.

[45]  H. Kotaki,et al.  Prediction of midazolam-CYP3A inhibitors interaction in the human liver from in vivo/in vitro absorption, distribution, and metabolism data. , 2001, Drug metabolism and disposition: the biological fate of chemicals.

[46]  O. Wisén,et al.  Caffeine metabolism in patients with chronic liver disease. , 1995, Scandinavian journal of clinical and laboratory investigation.

[47]  D. Mansuy,et al.  Substrate selectivity of human cytochrome P450 2C9: importance of residues 476, 365, and 114 in recognition of diclofenac and sulfaphenazole and in mechanism-based inactivation by tienilic acid. , 2003, Archives of biochemistry and biophysics.

[48]  D. Nicoll-Griffith,et al.  In Vitro Models for Studying Induction of Cytochrome P450 Enzymes , 2001, Drug-Drug Interactions.

[49]  D. Greenblatt,et al.  Differential Impairment of Triazolam and Zolpidem Clearance by Ritonavir , 2000, Journal of acquired immune deficiency syndromes.

[50]  Jiunn H. Lin,et al.  CYP Induction-Mediated Drug Interactions: in Vitro Assessment and Clinical Implications , 2006, Pharmaceutical Research.

[51]  A. Rettie,et al.  Mechanism-based inactivation of human cytochrome P450 1A2 by furafylline: detection of a 1:1 adduct to protein and evidence for the formation of a novel imidazomethide intermediate. , 1998, Biochemistry.

[52]  M. Eichelbaum,et al.  Inhibition of human CYP2B6 by N,N',N''-triethylenethiophosphoramide is irreversible and mechanism-based. , 2005, Biochemical pharmacology.

[53]  René H. Levy,et al.  Metabolic Drug Interactions , 2000 .

[54]  T. Maurer,et al.  Use of Immortalized Human Hepatocytes to Predict the Magnitude of Clinical Drug-Drug Interactions Caused by CYP3A4 Induction , 2006, Drug Metabolism and Disposition.

[55]  Yuichi Sugiyama,et al.  Quantitative Prediction of In Vivo Drug-Drug Interactions from In Vitro Data Based on Physiological Pharmacokinetics: Use of Maximum Unbound Concentration of Inhibitor at the Inlet to the Liver , 2000, Pharmaceutical Research.

[56]  R. Dal-Ré,et al.  Lack of Pharmacologic Interaction Between Paroxetine and Alprazolam at Steady State in Healthy Volunteers , 2004, Journal of clinical psychopharmacology.