Archives of Toxicology

, Volume 88, Issue 7, pp 1439–1449 | Cite as

A testing strategy to predict risk for drug-induced liver injury in humans using high-content screen assays and the ‘rule-of-two’ model

  • Minjun Chen
  • Chun-Wei Tung
  • Qiang Shi
  • Lei Guo
  • Leming Shi
  • Hong Fang
  • Jürgen Borlak
  • Weida Tong
In vitro systems

Abstract

Drug-induced liver injury (DILI) is a major cause of drug failures in both the preclinical and clinical phase. Consequently, improving prediction of DILI at an early stage of drug discovery will reduce the potential failures in the subsequent drug development program. In this regard, high-content screening (HCS) assays are considered as a promising strategy for the study of DILI; however, the predictive performance of HCS assays is frequently insufficient. In the present study, a new testing strategy was developed to improve DILI prediction by employing in vitro assays that was combined with the RO2 model (i.e., ‘rule-of-two’ defined by daily dose ≥100 mg/day & logP ≥3). The RO2 model was derived from the observation that high daily doses and lipophilicity of an oral medication were associated with significant DILI risk in humans. In the developed testing strategy, the RO2 model was used for the rational selection of candidates for HCS assays, and only the negatives predicted by the RO2 model were further investigated by HCS. Subsequently, the effects of drug treatment on cell loss, nuclear size, DNA damage/fragmentation, apoptosis, lysosomal mass, mitochondrial membrane potential, and steatosis were studied in cultures of primary rat hepatocytes. Using a set of 70 drugs with clear evidence of clinically relevant DILI, the testing strategy improved the accuracies by 10 % and reduced the number of drugs requiring experimental assessment by approximately 20 %, as compared to the HCS assay alone. Moreover, the testing strategy was further validated by including published data (Cosgrove et al. in Toxicol Appl Pharmacol 237:317–330, 2009) on drug-cytokine-induced hepatotoxicity, which improved the accuracies by 7 %. Taken collectively, the proposed testing strategy can significantly improve the prediction of in vitro assays for detecting DILI liability in an early drug discovery phase.

Keywords

Drug-induced liver injury DILI High-content screening assay Primary rat hepatocytes Drug safety assessment 

Supplementary material

204_2014_1276_MOESM1_ESM.pdf (48 kb)
Supplementary material 1 (PDF 48 kb)
204_2014_1276_MOESM2_ESM.pdf (25 kb)
Supplementary material 2 (PDF 25 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg (outside the USA) 2014

Authors and Affiliations

  • Minjun Chen
    • 1
  • Chun-Wei Tung
    • 1
    • 5
  • Qiang Shi
    • 2
  • Lei Guo
    • 3
  • Leming Shi
    • 6
  • Hong Fang
    • 4
  • Jürgen Borlak
    • 7
  • Weida Tong
    • 1
  1. 1.Division of Bioinformatics and Biostatistics, National Center for Toxicological ResearchUS Food and Drug AdministrationJeffersonUSA
  2. 2.Division of Systems Biology, National Center for Toxicological ResearchUS Food and Drug AdministrationJeffersonUSA
  3. 3.Division of Biochemical Toxicology, National Center for Toxicological ResearchUS Food and Drug AdministrationJeffersonUSA
  4. 4.Office of Scientific Coordination, National Center for Toxicological ResearchUS Food and Drug AdministrationJeffersonUSA
  5. 5.School of PharmacyKaohsiung Medical UniversityKaohsiungTaiwan
  6. 6.School of PharmacyFudan UniversityShanghaiChina
  7. 7.Center of Pharmacology and ToxicologyHannover Medical SchoolHannoverGermany

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