A NAMs-based framework for screening the endocrine-disrupting potential of plastic additives using cross-species...

The increasing global production of plastics has raised concerns about additive chemicals that can leach from polymer matrices and lead to widespread human and environmental exposure. While regulatory attention to endocrine-disrupting chemicals (EDCs) and their association with plastic additives has increased in recent years, hazard information for numerous plastic additives remains limited. Here, we applied an integrated new approach methodologies (NAMs)-based framework combining in silico molecular docking and in vivo Caenorhabditis elegans assays to efficiently screen the endocrine-disrupting potential of plastic additives. Molecular docking of 162 EU REACH-registered additives was performed against human estrogen and androgen receptors (ERα and AR), followed by cross-species docking using AlphaFold-predicted homology models of the C. elegans nuclear receptors (NHR-14 and NHR-69). Chemicals showing strong binding interactions across species were further examined through a systematic literature review focused on ER- and AR-mediated effects. By integrating evidence, 1,3-diphenylpropane-1,3-dione (DBM) and 2-(benzotriazol-2-yl)-4-methylphenol (UV-P) were identified as potential ER and AR disruptors and were subsequently validated through reproductive toxicity assays using C. elegans wild-type N2 and loss-of-function mutant strains (nhr-14 and nhr-69). This integrated approach provides a practical NAMs-based prioritization strategy by linking molecular interactions to phenotypic outcomes. By positioning C. elegans as a biological bridge between human toxicology and ecotoxicology, the framework supports cross-species hazard characterization within a One Health perspective.