Toxicity assessments of nonsteroidal anti-inflammatory drugs in isolated mitochondria, rat hepatocytes, and zebrafish show good concordance across chemical classes

• Published in: Toxicology and applied pharmacology Vol. 272; no. 2; pp. 272 - 280
• Main Authors: Nadanaciva, Sashi, Aleo, Michael D., Strock, Christopher J., Stedman, Donald B., Wang, Huijun, Will, Yvonne
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 15.10.2013; Elsevier
• Subjects: 60 APPLIED LIFE SCIENCES; ACETYLSALICYLIC ACID; Animals; Anti-Inflammatory Agents, Non-Steroidal - chemistry; Anti-Inflammatory Agents, Non-Steroidal - toxicity; Biological and medical sciences; Cell Survival - drug effects; Cells, Cultured; CONCENTRATION RATIO; Danio rerio; Drug Evaluation, Preclinical - methods; Drug toxicity and drugs side effects treatment; Embryo, Nonmammalian - drug effects; Embryo, Nonmammalian - pathology; ENDOPLASMIC RETICULUM; Freshwater; Hepatocytes - drug effects; Hepatocytes - enzymology; Hepatocytes - metabolism; Hepatocytes - pathology; High content imaging; INFLAMMATION; LIVER; LIVER CELLS; Liver toxicity; Male; Medical sciences; Membrane Potential, Mitochondrial - drug effects; METABOLITES; Miscellaneous (drug allergy, mutagens, teratogens...); MITOCHONDRIA; Mitochondria, Liver - drug effects; Mitochondria, Liver - enzymology; Mitochondria, Liver - metabolism; Mitochondria, Liver - pathology; Mitochondrial toxicity; MORPHOLOGY; No-Observed-Adverse-Effect Level; NSAIDS (non-steroidal anti-inflammatories); Oxygen Consumption - drug effects; Pharmacology. Drug treatments; Primary Cell Culture; RATS; RESPIRATION; SAFETY; TOXICITY; Toxicity Tests - methods; VIABILITY; Zebrafish; Zebrafish - embryology; GI; LOEC; AC50; NSAID; Zebrafish; Mitochondrial toxicity; COX; ER; UC50; ROR; IC50; NSAIDS (non-steroidal anti-inflammatories); NOEC; AERS; Liver toxicity; High content imaging; Evaluation; Rat; Digestive system; Toxicity; Chemical compound; Liver; Rodentia; In vitro; Non steroidal antiinflammatory agent; Vertebrata; Brachydanio rerio; Mitochondria; Mammalia; Pisces; Animal; Concordance; Content; Medical imagery; In vitro in vivo correlation; the concentration at which 50% inhibition occurred in State 3 respiration; UC; IC; adverse event reporting system; AC; the concentration at which 50% uncoupling occurred in State 2 respiration; cyclooxygenase; gastrointestinal; no observed effect concentration; half-maximal activity concentration; lowest observed effect concentration; endoplasmic reticulum; nonsteroidal anti-inflammatory drug; reporting odds ratio
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2013.06.019

To reduce costly late-stage compound attrition, there has been an increased focus on assessing compounds in in vitro assays that predict attributes of human safety liabilities, before preclinical in vivo studies are done. Relevant questions when choosing a panel of assays for predicting toxicity are (a) whether there is general concordance in the data among the assays, and (b) whether, in a retrospective analysis, the rank order of toxicity of compounds in the assays correlates with the known safety profile of the drugs in humans. The aim of our study was to answer these questions using nonsteroidal anti-inflammatory drugs (NSAIDs) as a test set since NSAIDs are generally associated with gastrointestinal injury, hepatotoxicity, and/or cardiovascular risk, with mitochondrial impairment and endoplasmic reticulum stress being possible contributing factors. Eleven NSAIDs, flufenamic acid, tolfenamic acid, mefenamic acid, diclofenac, meloxicam, sudoxicam, piroxicam, diflunisal, acetylsalicylic acid, nimesulide, and sulindac (and its two metabolites, sulindac sulfide and sulindac sulfone), were tested for their effects on (a) the respiration of rat liver mitochondria, (b) a panel of mechanistic endpoints in rat hepatocytes, and (c) the viability and organ morphology of zebrafish. We show good concordance for distinguishing among/between NSAID chemical classes in the observations among the three approaches. Furthermore, the assays were complementary and able to correctly identify “toxic” and “non-toxic” drugs in accordance with their human safety profile, with emphasis on hepatic and gastrointestinal safety. We recommend implementing our multi-assay approach in the drug discovery process to reduce compound attrition. •NSAIDS cause liver and GI toxicity.•Mitochondrial uncoupling contributes to NSAID liver toxicity.•ER stress is a mechanism that contributes to liver toxicity.•Zebrafish and cell based assays are complimentary.