In vitro approach to assess the potential for risk of idiosyncratic adverse reactions caused by candidate drugs.

Idiosyncratic adverse drug reactions (IADRs) in humans can result in a broad range of clinically significant toxicities leading to attrition during drug development as well as postlicensing withdrawal or labeling. IADRs arise from both drug and patient related mechanisms and risk factors. Drug related risk factors, resulting from parent compound or metabolites, may involve multiple contributory mechanisms including organelle toxicity, effects related to compound disposition, and/or immune activation. In the current study, we evaluate an in vitro approach, which explored both cellular effects and covalent binding (CVB) to assess IADR risks for drug candidates using 36 drugs which caused different patterns and severities of IADRs in humans. The cellular effects were tested in an in vitro Panel of five assays which quantified (1) toxicity to THLE cells (SV40 T-antigen-immortalized human liver epithelial cells), which do not express P450s, (2) toxicity to a THLE cell line which selectively expresses P450 3A4, (3) cytotoxicity in HepG2 cells in glucose and galactose media, which is indicative of mitochondrial injury, (4) inhibition of the human bile salt export pump, BSEP, and (5) inhibition of the rat multidrug resistance associated protein 2, Mrp2. In addition, the CVB Burden was estimated by determining the CVB of radiolabeled compound to human hepatocytes and factoring in both the maximum prescribed daily dose and the fraction of metabolism leading to CVB. Combining the aggregated results from the in vitro Panel assays with the CVB Burden data discriminated, with high specificity (78%) and sensitivity (100%), between 27 drugs, which had severe or marked IADR concern, and 9 drugs, which had low IADR concern, we propose that this integrated approach has the potential to enable selection of drug candidates with reduced propensity to cause IADRs in humans.

[1]  J. Landy,et al.  Trends in hospital admissions for pulmonary embolism. , 2008, Clinical medicine.

[2]  M. Al-Omary,et al.  Late-Onset Rosiglitazone-Associated Acute Liver Failure in a Patient with Hodgkin's Lymphoma , 2008, The Annals of pharmacotherapy.

[3]  J. G. Kenna,et al.  Cell based approaches for evaluation of drug-induced liver injury. , 2010, Toxicology.

[4]  Cynthia A Afshari,et al.  Interference with bile salt export pump function is a susceptibility factor for human liver injury in drug development. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[5]  Steven D. Cohen,et al.  A comparative study of mouse liver proteins arylated by reactive metabolites of acetaminophen and its nonhepatotoxic regioisomer, 3'-hydroxyacetanilide. , 1995, Chemical research in toxicology.

[6]  Kuresh Youdim,et al.  Application of PBPK modelling in drug discovery and development at Pfizer , 2012, Xenobiotica; the fate of foreign compounds in biological systems.

[7]  H. Zimmerman,et al.  Cholestatic hepatic injury related to warfarin exposure. , 1986, Archives of internal medicine.

[8]  H. Hosomi,et al.  Development of a Highly Sensitive Cytotoxicity Assay System for CYP3A4-Mediated Metabolic Activation , 2011, Drug Metabolism and Disposition.

[9]  Peter Greaves,et al.  First dose of potential new medicines to humans: how animals help , 2004, Nature Reviews Drug Discovery.

[10]  M. Sulkowski.  Drug-induced liver injury associated with antiretroviral therapy that includes HIV-1 protease inhibitors. , 2004, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[11]  Osamu Okazaki,et al.  A Zone Classification System for Risk Assessment of Idiosyncratic Drug Toxicity Using Daily Dose and Covalent Binding , 2009, Drug Metabolism and Disposition.

[12]  R. Andrade,et al.  Phenotypic characterization of idiosyncratic drug‐induced liver injury: The influence of age and sex , 2009, Hepatology.

[13]  Janet Clarke.  Mechanisms of adverse drug reactions to biologics. , 2010, Handbook of experimental pharmacology.

[14]  B. Andrews,et al.  New Technologies and Screening Strategies for Hepatotoxicity: Use of In Vitro Models , 2005, Toxicologic pathology.

[15]  J. Uetrecht,et al.  Immune-mediated adverse drug reactions. , 2009, Chemical research in toxicology.

[16]  I. Edwards,et al.  Adverse drug reactions: definitions, diagnosis, and management , 2000, The Lancet.

[17]  A Semi-Automated Method for Measuring White Matter Hyperintensity , 2013 .

[18]  G. Aithal.  Diclofenac-induced liver injury: a paradigm of idiosyncratic drug toxicity , 2004, Expert opinion on drug safety.

[19]  D. Keppler,et al.  The apical conjugate efflux pump ABCC2 (MRP2) , 2007, Pflügers Archiv - European Journal of Physiology.

[20]  A. Groen,et al.  Genetic defects in hepatobiliary transport. , 2002, Biochimica et biophysica acta.

[21]  J. Lacher,et al.  Zomepirac-induced renal failure. , 1983, Archives of internal medicine.

[22]  T. Y. Chan,et al.  Aminopyrine‐Induced Blood Dyscrasias—Still a Problem in Many Parts of the World , 1996, Pharmacoepidemiology and drug safety.

[23]  W. Humphreys,et al.  In vitro screening of 50 highly prescribed drugs for thiol adduct formation--comparison of potential for drug-induced toxicity and extent of adduct formation. , 2009, Chemical research in toxicology.

[24]  J. Burgunder,et al.  Liver injury due to verapamil. , 1988, Hepato-gastroenterology.

[25]  Mechanisms of Drug Hepatotoxicity in Man: Novel Insights Provided by the THLE-CYP Cell Panel , 2009 .

[26]  R. Ulrich,et al.  Idiosyncratic toxicity: a convergence of risk factors. , 2007, Annual review of medicine.

[27]  M. Pirmohamed.  Pharmacogenetics of idiosyncratic adverse drug reactions. , 2010, Handbook of experimental pharmacology.

[28]  S. Davies,et al.  Fatal acute fulminant liver failure due to clozapine: a case report and review of clozapine-induced hepatotoxicity. , 1997, Gastroenterology.

[29]  Dennis A. Smith,et al.  Multiple factors govern the association between pharmacology and toxicity in a class of drugs: toward a unification of class effect terminology. , 2011, Chemical research in toxicology.

[30]  M. Beconi,et al.  A semi-automated method for measuring the potential for protein covalent binding in drug discovery. , 2005, Journal of pharmacological and toxicological methods.

[31]  Chandan Saha,et al.  Relationship between daily dose of oral medications and idiosyncratic drug‐induced liver injury: Search for signals , 2008, Hepatology.

[32]  M. Monshouwer,et al.  An in vitro approach to detect metabolite toxicity due to CYP3A4-dependent bioactivation of xenobiotics. , 2005, Toxicology.

[33]  P. Ganey,et al.  Intrinsic versus Idiosyncratic Drug-Induced Hepatotoxicity—Two Villains or One? , 2010, Journal of Pharmacology and Experimental Therapeutics.

[34]  Yan Li,et al.  Risk assessment and mitigation strategies for reactive metabolites in drug discovery and development. , 2011, Chemico-biological interactions.

[35]  O. Yokosuka,et al.  Fulminant hepatic failure associated with benzbromarone treatment: A case report , 2002, Journal of Gastroenterology and Hepatology.

[36]  Jiri Aubrecht,et al.  Predicting safety toleration of pharmaceutical chemical leads: cytotoxicity correlations to exploratory toxicity studies. , 2010, Toxicology letters.

[37]  William M. Lee,et al.  Drug‐induced acute liver failure: Results of a U.S. multicenter, prospective study , 2010, Hepatology.

[38]  K. Jadallah,et al.  Acute hepatocellular-cholestatic liver injury after olanzapine therapy. , 2003, Annals of internal medicine.

[39]  T. Baillie,et al.  Drug-protein adducts: an industry perspective on minimizing the potential for drug bioactivation in drug discovery and development. , 2004, Chemical research in toxicology.

[40]  A. Daly.  Using genome-wide association studies to identify genes important in serious adverse drug reactions. , 2012, Annual review of pharmacology and toxicology.

[41]  P. Lurie,et al.  Case series of liver failure associated with rosiglitazone and pioglitazone , 2009, Pharmacoepidemiology and drug safety.

[42]  B. Dubois.  Using genome-wide association studies to better understand multiple sclerosis , 2012 .

[43]  R. Shepherd,et al.  Ibuprofen-Induced Liver Injury in an Adolescent Athlete , 2009, Clinical pediatrics.

[44]  R. Riley,et al.  Functional Consequences of Active Hepatic Uptake on Cytochrome P450 Inhibition in Rat and Human Hepatocytes , 2008, Drug Metabolism and Disposition.

[45]  B. Luxon,et al.  Nefazodone-Induced Liver Failure: Report of Three Cases , 1999, Annals of Internal Medicine.

[46]  Yvonne Will,et al.  The significance of mitochondrial toxicity testing in drug development. , 2007, Drug discovery today.

[47]  M. Roma,et al.  Hepatocellular transport in acquired cholestasis: new insights into functional, regulatory and therapeutic aspects. , 2008, Clinical science.

[48]  J. G. Kenna,et al.  In Vitro Inhibition of the Bile Salt Export Pump Correlates with Risk of Cholestatic Drug-Induced Liver Injury in Humans , 2012, Drug Metabolism and Disposition.

[49]  A. Kalgutkar,et al.  Can in vitro metabolism-dependent covalent binding data distinguish hepatotoxic from nonhepatotoxic drugs? An analysis using human hepatocytes and liver S-9 fraction. , 2009, Chemical research in toxicology.

[50]  R. Sandler.  Anaphylactic reactions to zomepirac. , 1985, Annals of emergency medicine.

[51]  A. Baumann.  Nonclinical development of biopharmaceuticals. , 2009, Drug discovery today.

[52]  Ian D. Wilson,et al.  Managing the challenge of chemically reactive metabolites in drug development , 2011, Nature Reviews Drug Discovery.

[53]  R Scott Obach,et al.  Can in vitro metabolism-dependent covalent binding data in liver microsomes distinguish hepatotoxic from nonhepatotoxic drugs? An analysis of 18 drugs with consideration of intrinsic clearance and daily dose. , 2008, Chemical research in toxicology.

[54]  Masashi Yabuki,et al.  Evaluation of the Potential for Drug-Induced Liver Injury Based on in Vitro Covalent Binding to Human Liver Proteins , 2009, Drug Metabolism and Disposition.

[55]  B. H. Ch. Stricker,et al.  Drug-induced Hepatic Injury , 1985 .

[56]  Robin Christensen,et al.  Efficacy and safety of the weight-loss drug rimonabant: a meta-analysis of randomised trials , 2007, The Lancet.

[57]  A. Stepan,et al.  Structural alert/reactive metabolite concept as applied in medicinal chemistry to mitigate the risk of idiosyncratic drug toxicity: a perspective based on the critical examination of trends in the top 200 drugs marketed in the United States. , 2011, Chemical research in toxicology.

[58]  J. Duan,et al.  A high throughput in vitro mrp2 assay to predict in vivo biliary excretion , 2012, Xenobiotica; the fate of foreign compounds in biological systems.

[59]  P. Meier,et al.  Functional expression of the canalicular bile salt export pump of human liver. , 2002, Gastroenterology.

[60]  K. Jemnitz,et al.  ABCC2/Abcc2: a multispecific transporter with dominant excretory functions , 2010, Drug metabolism reviews.

[61]  L. James,et al.  Mechanisms of acetaminophen-induced liver necrosis. , 2010, Handbook of experimental pharmacology.

[62]  B. Stieger,et al.  Genetic variations of bile salt transporters as predisposing factors for drug-induced cholestasis, intrahepatic cholestasis of pregnancy and therapeutic response of viral hepatitis , 2011, Expert opinion on drug metabolism & toxicology.

[63]  D. Wysowski,et al.  Adverse drug event surveillance and drug withdrawals in the United States, 1969-2002: the importance of reporting suspected reactions. , 2005, Archives of internal medicine.

[64]  I. E. El Hajj,et al.  Celecoxib-induced cholestatic liver failure requiring orthotopic liver transplantation. , 2009, World journal of gastroenterology.

[65]  Yuan Chen,et al.  Application of modern drug metabolism structure determination tools and assays to the in vitro metabolism of imiloxan. , 2010, Drug metabolism letters.

[66]  Josip Car,et al.  Trends in hospital admissions for adverse drug reactions in England: analysis of national hospital episode statistics 1998–2005 , 2007, BMC clinical pharmacology.