Minjun Chen, PhD Principal Investigator National Center for Toxicological Research, FDA

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Minjun Chen, PhD Principal Investigator National Center for Toxicological Research, FDA Click to view Biosketch and Presentation Abstract or page down to review presentation

The Rule of 2 Do Drug Properties Predict Drug-Induced Liver Injury (DILI)? Minjun Chen, Ph.D. Division of Bioinformatics and Biostatics, The US FDA s National Center for Toxicological Research Minjun.chen@fda.hhs.gov Views expressed in this presentation are those of the presenter and not necessarily those of the U.S. FDA. March 19 th, 2015

Chen M, Suzuki A, Borlak, Lucena M, Andrade R, (2015) J Hepatol, submitted

Translational Challenges in DILI Risk Assessment Preclinical toxicology often failed to predict DILI risk in humans High-dosing of healthy animals test identified < 50% DILI liability in humans Conventional in vitro assays have very limited prediction for DILI risk assessment No biomarker is available to identify the susceptible patients prior to drug treatment Kaplowitz N (2005) Nat Rev Drug Disov,4:489-99; Avigan M (2014) Seminars in Liver Disease,34:215-226; Chen M, Borlak J, Tong W (2014) Expert Rev Gastroenterol Hepatol, 8: 721-23.

Assess DILI risk of new chemicals Liver Toxicity Knowledge Base (LTKB) Search LTKB @ Google Link to external resources, e.g., PubMed, Wiki, LiverTox Chemical structure & similarity search Excel-like spreadsheet Query interface Drug specific data 4

- Chemical structure (SAR/QSAR) - Daily dose (Cmax) - Lipophilicity - Reactive Metabolites - P450 activities Therapeutic uses - Pharmacological class ATC FDA classification - Side effects - Japanese TGX Project >130 drugs 4 testing systems Multi-doses and times >20,000 arrays - DrugMatrix ~700 chemicals ~5000 arrays - Others (e.g., in-house) >40K arrays (>500 drugs) - In house in vitro data Human primary hepatocytes Rat primary hepatocytes HepG2 - Tox21 and ToxCast - Others In vitro assay (>200 drugs) ~3000 Side effects DILI risk - DILI types - DILI ontology - DILI annotation based on FDA drug label - Severe DILI - Case reports Chen M, Zhang J, Wang Y, Liu Z, Kelly R, Zhou G, Fang H, Borlak J, Tong W (2013) Clin Pharmacol Ther, 93 : 409-412

How to Assess DILI Risk in Humans for a Drug? Three attributes of a drug are important for its DILI assessment: Causality: was liver injury caused by drug or other cause Incidence: how many case reports are considered significant Severity: elevated ALT; Hy s law; disability and hospitalization, liver failure; liver transplantation or death Risk = (How likely) x (How many) x (How severe) This is opinion based!!! 6

Drug Labeling is not Perfect but Probably the Best and Mostly Consistent Information We Have Drug labeling is not perfect Opinion based, not based on pre-defined criteria Not according to a consistent or scientifically justified master plan Guilty by Association carryover from other drugs Adverse Reactions: This section must list the adverse reactions that occur with the drug and with drugs in the same pharmacologically active and chemically related class, if applicable Can we use drug labeling to identify more dangerous drugs? The question is how to mitigate the inherent defects in labeling! 7

Chen M, Vijay V, Shi Q, Liu Z, Fang H, Tong W (2011) Drug Discov Today, 16: 697-703 Drug labels 8

The Rule of 2 ( RO2 ): High Lipophilicity (logp 3) + High Daily Dose (DD 100 mg) Predicts DILI 10 8 6 4 2 Most DILI-concern No DILI-concern DILI logp 0-2 -4-6 -8 - Observed in 164 drugs - Verified by 179 drugs - Demonstrated on 5 drug pairs - Applied to co-medication -10 0.001 0.01 0.1 1 10 100 1000 10000 100000 Daily dose (mg) Chen M, Borlak J, Tong W (2013) Hepatology, 58(1): 388-396

Assess RO2 Model by Drug Pairs Compound Name Daily dose (mg/day) AlogP RO2 test Label Section DILI Concern Macitentan 10 3.5 Negative Ambrisentan 7.5 3.66 Negative Warning & Precaution Adverse Reaction Less-DILI-Concern Less-DILI-Concern Bosentan 250 3.95 Positive Box Warning Most-DILI-Concern Sitaxsentan 100 3.7 Positive Withdrawn Most-DILI-Concern Compound Daily dose Name (mg/day) AlogP RO2 test Label Section DILI Concern Zolpidem 10 3.36 Negative Adverse reaction Less-DILI-concern Alpidem 150 5.46 Positive Withdrawn Most-DILI-Concern Piglitazone 6 3.91 Negative Warnings & Precautions Less-DILI-concern Rosiglitazone 30 3.36 Negative Warnings & Precautions Less-DILI-concern Troglitazone 400 5.1 Positive Withdrawn Most-DILI-Concern 10

More of RO2 Successful and Failed Cases Compound Name Daily dose (mg/day) AlogP RO2 test Label Section DILI Concern Entacapone 1000 1.66 Negative No mentioned No-DILI-concern Tolcapone 450 3.13 Positive Box warning Most-DILI-Concern Trazodone 300 2.42 Negative Adverse reaction Less-DILI-concern Nefazodone 400 4.42 Positive Withdrawn Most-DILI-Concern RO2 failed cases (30-35%sensitivity and 90-95%specificity) Compound Name Daily dose (mg/day) AlogP RO2 test DILI concern Comments Trovafloxacin 200-0.91 False Negative Withdrawn Ximelagatran 48 1.16 False Negative Withdrawn Aliskiren 150 3.32 False Positive No-DILI-concern Bioavailability (2.6%) Diphenhydram ine 300 3.38 False Positive No-DILI-concern Antihistamine, Modulate immune 11

RO2 Validation by FDA-Approved Oral Medications 748 oral drugs RO2 positive RO2 negative 168 most-dili-concern Sensitivity = 72/168 (43%) 193 no-dili-concern Specificity = 1-11/193 (94%) 387 less-dili-concern RO2 pos% = 50/387 (13%) 95 182 72 11 337 50 * Withdrawn RO2 pos%= 51% Box warning RO2 pos%= 38% Warnings RO2 pos%= 40% 24 18 54 25 11 36 Immune modulation: diphenhydramine, orphenadrine, chlorcyclizine, flavoxate, benzogetamine, mifepristone, pentazocine, ursodeoxycholic acid Poor bioavailability(%): aliskiren (3%) Unknown: retigabine, tapentadol * 23 of 50 RO2 positives were assigned as DILI positives by at least one literature 12

Selected Failed Drug Development Due to Hepatotoxicity Compound Daily Dose Name (mg/day) AlogP RO2 test Safety Concerns NP260 3000* 5 Positive Liver toxicity were found in beagle dog Casopitant 150 4.73 Positive Liver toxicity were found in Phase III patients Tasosartan 200 3.051 Positive Phase III clinical trials showed elevated transaminases Fiduxosin 100 4.215 Positive Discontinued due to liver toxicity Alaproclate 100* 2.695 Negative Observed liver complications in rodent studies Pafuramidine 100 2.77 Negative Liver abnormalities identified in healthy volunteers Aplaviroc 1200 1.386 Negative Severe hepatotoxicity observed in Phase III studies Pralnacasan 300 0.132 Negative Liver abnormality observed at one species of animals Fialuridine 15-1.228 Negative Unexpected fulminant liver failure in patients * Equivalent human dose Call for the sharing of the failed drugs!

Improve RO2 by Integrating High-Content Screening (HCS) Assays Human hepatotoxicity # drugs Model Test result Positive (N=49) Negative (N=21) Accuracy Sensitivity Specificity requiring HCS test RO2-HCS Positive 27 1 Negative 22 20 67% 55% 95% 56 HCS Positive 19 0 Negative 30 21 57% 39% 100% 70 Positive 13 1 RO2 47% 27% Negative 36 20 95% N/A HCS RO2 13 6 7 Total positives (N=49) Chen M, Tung C, Shi Q, Guo L, Shi L, Fang H, Borlak J, Tong W, (2014) Arch Toxicol, 88:1439-49 14

Drug Properties or Host Factors? It is dose! It is mitochondria! DILI It is genome! It is HLA! It is immune! It is reactive metabolites! Finally, they stop talking, start listening and collaborate to "see" the full elephant. 15

Take-home Messages Along with host factors, drug properties also contribute to the prediction of DILI LTKB provides a centralized repository of diverse DILI-related drug properties data. Several predictive models were developed The RO2 has added value in identifying idiosyncratic DILI In vitro assays can be enhanced by integrating the RO2 Improvements have been made, but still a long way to go. 16

Acknowledgements LTKB interest group: Eileen E Navarro Almario John Senior Marc Stone Tina Burgrass Ruyi He Shashi Amur Jane Bai Crentsil Victor Andrew Mulberg Mark Avigan Non-FDA collaborators: Jurgen Borlak (Hannover Medical School, Germany) Ayako Suzuki (UAMS, US) Raul Andrade & Maribel Lucena (Spanish hepatotoxicity registry) Xiaowei Xu (UALR, US) NCTR: Weida Tong Gerry Zhou Ke Yu Jie Zhang Yuping Wang Hong Fang Huixiao Hong Joshua Xu Halil Bisgin Zhichao Liu Chuchu Hu FDA Supports: Critical Path Initiatives Office of Women s Health (OWH) Chief Scientist Challenge Grant Office of International Program 17