Relationship between Passive Permeability, Efflux, and Predictability of Clearance from In Vitro Metabolic Intrinsic Clearance

In vitro intrinsic metabolic clearance (CLint) is used routinely for compound selection in drug discovery; however, in vitro CLint often underpredicts in vivo clearance (CL). Forty-one proprietary compounds and 16 marketed drugs were selected to determine whether permeability and efflux status could influence the predictability of CL from in vitro CLint obtained from liver microsomal and hepatocyte incubations. For many of the proprietary compounds examined, rat CL was significantly underpredicted using the well stirred model incorporating both fraction of unbound drug in blood and fraction of unbound drug in the microsomal or hepatocyte incubation. Further analysis revealed that the accuracy of the prediction was differentiated by permeability and P-glycoprotein- (P-gp) and mouse breast cancer resistance protein (mBcrp)-mediated efflux. For proprietary compounds with passive permeability greater than 5 × 10−6 cm/s and efflux ratios less than 5 in both P-gp- and mBcrp-expressing cells, CLint provided reasonable prediction. The average -fold error (AFE) was 1.8 for rat liver microsomes (RLMs) and 2.3 for rat hepatocytes. In contrast, CL was dramatically underpredicted for compounds with passive permeability less than 5 × 10−6 cm/s; AFEs of 54.4 and 29.2 were observed for RLM and rat hepatocytes, respectively. In vivo CL was also underpredicted for compounds that were good efflux substrates (permeability >5 × 10−6 cm/s). The AFEs were 7.4 and 8.1 for RLM and rat hepatocytes, respectively. A similar relationship between permeability, efflux status, and human CL prediction reported in the literature was observed for 16 marketed drugs. These data show that permeability and efflux status are determinants for the predictability of CL from in vitro metabolic CLint.

[1]  J. Lima,et al.  Absorption of montelukast is transporter mediated: a common variant of OATP2B1 is associated with reduced plasma concentrations and poor response , 2009, Pharmacogenetics and genomics.

[2]  J. Schellens,et al.  The effect of P-gp (Mdr1a/1b), BCRP (Bcrp1) and P-gp/BCRP inhibitors on the in vivo absorption, distribution, metabolism and excretion of imatinib , 2009, Investigational New Drugs.

[3]  S. Bates,et al.  ABCG2: a perspective. , 2009, Advanced drug delivery reviews.

[4]  J. Houston,et al.  Reliability of human cryopreserved hepatocytes and liver microsomes as in vitro systems to predict metabolic clearance , 2008 .

[5]  Bo Feng,et al.  In Vitro P-glycoprotein Assays to Predict the in Vivo Interactions of P-glycoprotein with Drugs in the Central Nervous System , 2008, Drug Metabolism and Disposition.

[6]  B. Stieger,et al.  Bosentan Is a Substrate of Human OATP1B1 and OATP1B3: Inhibition of Hepatic Uptake as the Common Mechanism of Its Interactions with Cyclosporin A, Rifampicin, and Sildenafil , 2007, Drug Metabolism and Disposition.

[7]  J Brian Houston,et al.  Evaluation of Cryopreserved Human Hepatocytes as an Alternative in Vitro System to Microsomes for the Prediction of Metabolic Clearance , 2007, Drug Metabolism and Disposition.

[8]  Yuichi Sugiyama,et al.  Transporters as a determinant of drug clearance and tissue distribution. , 2006, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[9]  R. Austin,et al.  A UNIFIED MODEL FOR PREDICTING HUMAN HEPATIC, METABOLIC CLEARANCE FROM IN VITRO INTRINSIC CLEARANCE DATA IN HEPATOCYTES AND MICROSOMES , 2005, Drug Metabolism and Disposition.

[10]  Mikko Niemi,et al.  Polymorphic Organic Anion Transporting Polypeptide 1B1 is a Major Determinant of Repaglinide Pharmacokinetics , 2005, Clinical pharmacology and therapeutics.

[11]  Mariël Brok,et al.  Imatinib mesylate (STI571) is a substrate for the breast cancer resistance protein (BCRP)/ABCG2 drug pump. , 2004, Blood.

[12]  Lasse Murtomäki,et al.  Drug permeation in biomembranes: in vitro and in silico prediction and influence of physicochemical properties. , 2004, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[13]  Kiyomi Ito,et al.  Comparison of the Use of Liver Models for Predicting Drug Clearance Using in Vitro Kinetic Data from Hepatic Microsomes and Isolated Hepatocytes , 2004, Pharmaceutical Research.

[14]  D. Nakai,et al.  Evaluation of the protein binding ratio of drugs by a micro-scale ultracentrifugation method. , 2004, Journal of pharmaceutical sciences.

[15]  R. de Kanter,et al.  Prediction of whole-body metabolic clearance of drugs through the combined use of slices from rat liver, lung, kidney, small intestine and colon , 2004, Xenobiotica; the fate of foreign compounds in biological systems.

[16]  I. Bjørnsdottir,et al.  CYP2C8 and CYP3A4 are the principal enzymes involved in the human in vitro biotransformation of the insulin secretagogue repaglinide. , 2003, British journal of clinical pharmacology.

[17]  Yuichi Sugiyama,et al.  Utility of hepatocytes in predicting drug metabolism: comparison of hepatic intrinsic clearance in rats and humans in vivo and in vitro. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[18]  W. Elmquist,et al.  Distribution of STI-571 to the Brain Is Limited by P-Glycoprotein-Mediated Efflux , 2003, Journal of Pharmacology and Experimental Therapeutics.

[19]  Yuichi Sugiyama,et al.  Inhibition of Transporter-Mediated Hepatic Uptake as a Mechanism for Drug-Drug Interaction between Cerivastatin and Cyclosporin A , 2003, Journal of Pharmacology and Experimental Therapeutics.

[20]  A. Volz,et al.  Altered drug disposition of the platelet activating factor antagonist apafant in mdr1a knockout mice. , 2002, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[21]  Terry R Stouch,et al.  Progress in understanding the structure-activity relationships of P-glycoprotein. , 2002, Advanced drug delivery reviews.

[22]  William J Egan,et al.  Prediction of intestinal permeability. , 2002, Advanced drug delivery reviews.

[23]  L. Benet,et al.  Unmasking the dynamic interplay between intestinal P-glycoprotein and CYP3A4. , 2002, The Journal of pharmacology and experimental therapeutics.

[24]  J. Polli,et al.  Rational use of in vitro P-glycoprotein assays in drug discovery. , 2001, The Journal of pharmacology and experimental therapeutics.

[25]  Y. Sugiyama,et al.  Prediction of human hepatic clearance from in vivo animal experiments and in vitro metabolic studies with liver microsomes from animals and humans. , 2001, Drug metabolism and disposition: the biological fate of chemicals.

[26]  R. Obach,et al.  Prediction of human clearance of twenty-nine drugs from hepatic microsomal intrinsic clearance data: An examination of in vitro half-life approach and nonspecific binding to microsomes. , 1999, Drug metabolism and disposition: the biological fate of chemicals.

[27]  G. Hopfgartner,et al.  Absorption, excretion, and metabolism of the endothelin receptor antagonist bosentan in healthy male subjects. , 1999, Drug metabolism and disposition: the biological fate of chemicals.

[28]  R. Miller,et al.  Metabolic profiles of montelukast sodium (Singulair), a potent cysteinyl leukotriene1 receptor antagonist, in human plasma and bile. , 1997, Drug metabolism and disposition: the biological fate of chemicals.

[29]  J. Beijnen,et al.  Limited oral bioavailability and active epithelial excretion of paclitaxel (Taxol) caused by P-glycoprotein in the intestine. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Piet Borst,et al.  Substantial excretion of digoxin via the intestinal mucosa and prevention of long‐term digoxin accumulation in the brain by the mdrla P‐glycoprotein , 1996, British journal of pharmacology.

[31]  Tim Morris,et al.  Physiological Parameters in Laboratory Animals and Humans , 1993, Pharmaceutical Research.

[32]  P. Seglen.  Preparation of rat liver cells. 3. Enzymatic requirements for tissue dispersion. , 1973, Experimental cell research.

[33]  R. Evers,et al.  Importance of mechanistic drug metabolism studies in support of drug discovery: A case study with an N -sulfonylated dipeptide VLA-4 antagonist in rats. , 2008, Xenobiotica; the fate of foreign compounds in biological systems.

[34]  Mariël Brok,et al.  Imatinib mesylate ( STI 571 ) is a substrate for the breast cancer resistance protein ( BCRP ) / ABCG 2 drug pump , 2004 .

[35]  Leslie Z. Benet,et al.  Predicting Drug Disposition via Application of BCS: Transport/Absorption/ Elimination Interplay and Development of a Biopharmaceutics Drug Disposition Classification System , 2004, Pharmaceutical Research.

[36]  J. Tolan,et al.  MDCK (Madin-Darby canine kidney) cells: A tool for membrane permeability screening. , 1999, Journal of pharmaceutical sciences.

[37]  P. Seglen.  Preparation of rat liver cells. II. Effects of ions and chelators on tissue dispersion. , 1973, Experimental cell research.