Mechanism-based toxicity screening of organophosphate flame retardants using Tox21 assays and molecular docking analysis

As brominated flame retardants are phased out and regulations on their use become stricter, concerns over organophosphate flame retardants (OPFRs) have increased due to their high production. In response, this study aimed to screen the potential toxicity of emerging OPFRs using in vitro Tox21 assays and in silico molecular docking analysis. For 48 OPFRs collected from the literature, we investigated their bioactivity with human nuclear receptors using Tox21 data, focusing on pathways related to endocrine disruption (ERs, AR), stress response (GR), energy homeostasis (PPARs, FXR), and detoxification (PXR, CAR). For OPFRs not tested in Tox21 assays, molecular docking simulations were performed to predict binding potential. Results showed that CAR/PXR and FXR had relatively high reactivity with diverse OPFRs, indicating potential molecular initiating events (MIEs). Among the 48 OPFRs, 28 interacted with one or more receptors, suggesting they may act as potential stressors of adverse outcome pathways (AOPs) leading to various human diseases. Aryl- and halogenated-OPFRs displayed higher bioactivity compared to alkyl-OPFRs. Additionally, as the logKow value and carbon number of OPFRs increased, their interaction with nuclear receptors also increased. These structure- and physicochemistry-dependent bioactivities provide insights for designing safer OPFRs to avoid regrettable substitutions. Of these prioritized OPFRs, 13 showed low oral points-of-departure (POD) values under 100 mg/kg/day. In contrast, the other 15 OPFRs lacked sufficient data or exhibited less severe toxicity, despite being predicted to be of high concern in our analysis. Since several OPFRs are commonly used in consumer products that can lead to daily human exposure, we suggest that these OPFRs have the potential to reveal undisclosed effects and should therefore undergo further assessment.