Predicting acute oral toxicity using AcutoX: An animal product-free and metabolically relevant human cell-based test
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Abstract
AcutoX is a human in vitro test method for the evaluation of acute oral toxicity, developed using a library of 67 curated test chemicals. These chemicals cover a wide variety of chemistries, industrial sectors, rodent toxicities, and all EPA and GHS hazard categories. The test uses two different cytotoxicity endpoints (Neutral Red uptake and MTT metabolism), performed both in the presence and absence of a pooled human liver extract (S9), to produce four EC50 values. The EC50 values are used in prediction models to assign a “highly toxic” and “low toxicity” category for both EPA and GHS classification, which can be further refined to assign a hazard category. The binary “highly toxic” / “low toxicity” prediction model has an accuracy of 73.8% and 63.1% for EPA and GHS, respectively, with the subsequent hazard categorization offering a protective prediction (correct or higher category) in 90.0% and 93.3% of cases, respectively. Moreover, the AcutoX test can identify chemicals activated or detoxified by liver metabolism.
Plain language summary
AcutoX is a human-relevant laboratory test that can help to determine the toxicity of a chemical to human health if a chemical was to be ingested. The AcutoX test does not use animal models, or any components derived from animals, and contains a function that mimics human metabolism. Over 60 chemicals of varying degrees of known toxicity were run through the AcutoX test and the results were compared to widely available toxicity data that was obtained in animal models. The data comparisons revealed that the AcutoX test could correctly predict the safety of a significant number of chemicals.
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Alberga D, Trisciuzzi D, Mansouri K, et al. (2019). Prediction of Acute Oral Systemic Toxicity Using a Multifingerprint Similarity Approach. Toxicol Sci 167; 484-495. doi:10.1093/toxsci/kfy255
Clothier, R, Gomez-Lechon, MJ, Kinsner-Ovaskainen, et al. (2013). Comparative analysis of eight cytotoxicity assays evaluated within the ACuteTox Project. Toxicol In Vitro 27, 1347-1356. doi:10.1016/j.tiv.2012.08.015
Daniel AB, Choksi N, Abedini J et al. (2022). Data curation to support toxicity assessments using the Integrated Chemical Environment. Front Toxicol 3; 987848. doi:10.3389/ftox.2022.987848
Furukawa T, Naritomi Y, Tetsuka K, et al. (2014). Species differences in intestinal glucuronidation activities between humans, rats, dogs and monkeys. Xenobiotica 44, 205-16. doi:10.3109/00498254.2013.828362
Furness JB, Cottrell JJ, and Bravo DM. (2015). Comparative gut physiology symposium: comparative physiology of digestion. J Anim Sci 93, 485-91. doi:10.2527/jas.2014-8481
Hamm J, Allen D, Ceger P et al. (2021). Performance of the GHS mixtures equation for predicting acute oral toxicity. Regul Toxicol Pharmacol 125, 105007. doi:10.1016/j.yrtph.2021.105007
Hammer H, Schmidt F, Marx-Stoelting P, et al. (2021). Cross species analysis of hepatic cytochrome P450 and transport protein expression. Arch Toxicol 95, 117-33. doi:10.1007/s00204-020-02939-4
Kararli TT. (1995). Comparison of the gastrointestinal anatomy, physiology, and biochemistry of humans and commonly used laboratory animals. Biopharm Drug Dispos 16, 351-80. doi:10.1002/bdd.2510160502
Karmaus AL, Mansouri K, To KT et al. (2022). Evaluation of variability across rat acute oral systemic toxicity studies. Toxicol Sci 188, 34-47. doi:10.1093/toxsci/kfac042
Kogure K, Ishizaki M, Nemoto M, et al. (1999). A comparative study of the anatomy of rat and human livers. J Hepatobiliary Pancreat Surg 6, 171-5. doi:10.1007/s005340050101
Kojima H, Nakada T, Yagami AT et al. (2023). A step-by-step approach for assessing acute oral toxicity without animal testing for additives of quasi-drugs and cosmetic ingredients. Curr Res Toxicol 4; doi:10.1016/j.crtox.2022.100100
Mansouri K, Karmaus AL, Fitzpatrick J et al. (2021). CATMoS: Collaborative acute toxicity modeling suite. Environ Health Perspect 129, doi:10.1289/EHP8495. Erratum in: Environ Health Perspect 129, 79001.
Martignoni M, Groothuis GM, and de Kanter R. (2006). Species differences between mouse, rat, dog, monkey and human CYP-mediated drug metabolism, inhibition and induction. Expert Opin Drug Metab Toxicol. 2, 875-94. doi:10.1517/17425255.2.6.875
Movia D, Bruni-Favier S and Prina-Mello A. (2020). In vitro alternatives to acute inhalation toxicity studies in animal models - a perspective. Front Bioeng Biotechnol 8, 549. doi:0.3389/fbioe.2020.00549
Mukai M, Isobe T, Okada K, et al. (2015). Species and sex differences in propofol glucuronidation in liver microsomes of humans, monkeys, rats and mice. Pharmazie 70; 466-70.
Nishimuta H, Nakagawa T, Naruaki N, et al. (2013). Species differences in hepatic and intestinal metabolic activities for 43 human cytochrome P450 substrates between humans and rats or dogs. Xenobiotica 43, 948-55. doi:10.3109/00498254.2013.787155
OECD (2016). Guidance Document No. 237: Guidance Document on Considerations for Waiving or Bridging of Mammalian Acute Toxicity Tests. OECD Publishing, Paris.
OECD (2002a). Test No. 420: Acute oral toxicity - fixed dose procedure. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris. doi.org/10.1787/9789264070943-en.
OECD (2002b). Test No. 423: Acute oral toxicity - acute toxic class method. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris. doi.org/10.1787/9789264071001-en.
OECD (2010). Series on testing and assessment No. 129. Guidance document on using cytotoxicity tests to estimate starting doses for acute oral systemic toxicity tests. https://one.oecd.org/document/env/jm/mono(2010)20/en/pdf
OECD (2022). Test No. 425: Acute oral toxicity: up-and-down procedure. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris. doi:10.1787/9789264071049-en
OECD (2023). Guideline No. 497: Defined Approaches on Skin Sensitisation. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris. doi:10.1787/b92879a4-en
Prieto P, Kinsner-Ovaskainen S, Stanzel S, et al (2013). The value of selected in vitro and in silico methods to predict oral acute toxicity in a regulatory context: Results from the European Project ACuteTox. Toxicol In Vitro 27, 1357-1376. doi:10.1016/j.tiv.2012.07.013
Schrage A, Hempel K, Schulz M, et al. (2011). Refinement and reduction of acute oral toxicity testing: a critical review of the use of cytotoxicity data. Altern Lab Anim 39, 273-95. doi:10.1177/026119291103900311
Sjöström M, Kolman A, Clemedson C et al. (2008). Estimation of human blood LC50 values for use in modeling of in vitro-in vivo data of the ACuteTox project. Toxicol In Vitro 22, 1405-11. doi:10.1016/j.tiv.2008.04.017
Strickland J, Clippinger AJ, Brown J et al. (2018). Status of acute systemic toxicity testing requirements and data uses by U.S. regulatory agencies. Regul Toxicol Pharmacol 94, 183-196. doi:10.1016/j.yrtph.2018.01.022
Strickland J, Haugabrooks E, Allen DG, et al. (2023). International regulatory uses of acute systemic toxicity data and integration of new approach methodologies. Crit Rev Toxicol 53, 385-411. doi:10.1080/10408444.2023.2240852
Taberner PV and Pearce MJ. (1974) Hypothermic and toxic actions of 2–butyne-1, 4-diol and other related diols in the rat, J Pharm Pharmacol 26; 597–604. doi:10.1111/j.2042-7158.1974.tb10671.x
UN – United Nations (2023). Globally harmonized system of classification and labelling of chemicals. 10th revised edition. https://unece.org/transport/documents/2023/07/standards/ghs-rev10
van der Zalm AJ, Barroso J, Browne P et al. (2022). A framework for establishing scientific confidence in new approach methodologies. Arch Toxicol 96, 2865-2879. doi:10.1007/s00204-022-03365-4
Williams J (1928). The decomposition of hydrogen peroxide by liver catalase. J Gen Physiol 11, 309-37. doi:10.1085/jgp.11.4.309