Bioactivation of sitaxentan in liver microsomes, hepatocytes, and expressed human P450s with characterization of the glutathione conjugate by liquid chromatography tandem mass spectrometry
Abstract
Sitaxentan is a selective endothelin-A receptor antagonist that was marketed as Thelin in several European countries and Canada for pulmonary arterial hypertension. Sitaxentan was undergoing further clinical trials in the United States but due to four deaths and one case of liver transplantation from severe liver toxicity that appeared to be idiosyncratic in nature, it was withdrawn worldwide in December, 2010. Sitaxentan contains a 1,3-benzodioxole ring that undergoes enzymatic demethyleneation to an ortho-catechol metabolite that can further oxidize to a reactive ortho-quinone metabolite. Here, we report the detection and mass spectral characterization of a glutathione conjugate of this sitaxentan quinone reactive metabolite that was trapped in vitro using mouse, rat, dog, and human liver microsomes supplemented with NADPH and glutathione and that was also observed in rat and human hepatocytes. Using human liver microsomes, we also demonstrated that P450 3A4 undergoes time-dependent inhibition. Density functional calculations on the catechol metabolite of sitaxentan indicated that the reaction leading to the quinone was thermodynamically favorable with an enthalpy change of -6.3 kcal/mol. Using density functional methodology, we modeled the attack of glutathione on the quinone with an S-methyl thiolate anion which allowed us to predict, based on the difference in transition state energies, that the 2-position on the phenyl ring was more likely than the 5-position as the site of glutathione conjugation. Overall, our results demonstrated that sitaxentan is capable of facile formation of a reactive ortho-quinone metabolite capable of reacting with glutathione and may rationalize the idiosyncratic nature of the hepatotoxicity that led to its withdrawal.
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Sitaxentan caused no explicit liver injury in animal experiments, whereas hepatotoxicity was prominent in humans.
“Sitaxentan caused no explicit liver injury in animal experiments [8], whereas the hepatotoxicity was prominent in humans [4,5].”
From Machine Learning Based Toxicity Prediction: From Chemical Structural Description to Transcriptome Analysis (2018) · cites this paper as [4]
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- Authors
- Erve, J. C. L., Gauby, S., Maynard, J. W., Svensson, M. A., Tonn, G., & Quinn, K. P.
- Affiliations
- Department of Drug Metabolism and Pharmacokinetics, Elan Pharmaceuticals Inc., 180 Oyster Point Boulevard, South San Francisco, California 94080, United States
- Schrodinger (United States) (US)
- Corresponding author
- Erve, John C. L.
- Venue
- Chemical Research in Toxicology
- Publisher
- American Chemical Society (ACS)
- Published
- 2013-05-30
- Volume
- 26
- Issue
- 6
- Pages
- 926-936
- ISSN
- 0893-228X (ISSN-L)
- 1520-5010
- OpenAlex
- W2315575379
- Cited by
- 48
- References
- 42
- Citations by year
- 2026: 4 · 2025: 5 · 2024: 2 · 2023: 3 · 2022: 6 · 2021: 3 · 2020: 4 · 2019: 5 · 2018: 5 · 2017: 3 · 2016: 3 · 2015: 3 · 2014: 2
- FWCI
- 2.91
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Chemical Research in Toxicology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Bioactive Compounds and Antitumor Agents
- Keywords
- sitaxentan, metabolic activation, glutathione conjugates, endothelin A receptor antagonist, idiosyncratic hepatotoxicity, DFT calculations, reactive metabolites, drug withdrawal, liver microsomes, hepatocytes
- MeSH terms
- Animals, Biotransformation, Catechols, Chromatography, Liquid, Cytochrome P-450 Enzyme System, Dogs, Glutathione, Humans, Isoxazoles, Microsomes, Liver, Quantum Theory, Thiophenes, Benzoquinones, Hepatocytes, Mice, Rats, Tandem Mass Spectrometry, Cytochrome P-450 Enzyme Inhibitors
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