ZeroPM prioritization workshop presentation Data Integration and Risk Matrix

Keynote speaker – Elisabet Berggren •Deputy Head Systems Toxicology Unit, Joint Research Centre (JRC) - European Commission •European Union Reference Laboratory for Alternatives to Animal Testing (ECVAM) established to protect animals used for scientific purposes •Commission roadmap towards phasing out of animal testing in chemicals safety assessment •EPAA Designathon based on a future vision of chemicals safety assessment - Chemicals 2.0 •United Nations Globally Harmonised System (GHS) for Classification and Labelling of Chemicals •Future regulatory framework for chemicals in the European Union This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 101036756. Data Integration and Risk Matrix S. E. Escher, T. Gouin and T. Hamers Goal To develop a risk-based prioritization framework for substances qualified as Persistent and Mobile •Taking into account toxicological hazard and exposure •With a focus on human exposure through drinking water •As part of a tiered approach Starting points: •Based on new-approach methodologies (NAMs) •Outcomes to be visualized in a risk matrix RISK21, Embry et al. 2014 Crit Rev Toxicol 44: 6-16 Integrate “T” into grouping TriazolesTriazines Per-and polyfluoroalkyl substances (PFAS) TebuconazoleCyromazine PFOS Grouping based on biological and chemical Similarity - global approach usingToxCast data Tebuconazole Bitertanol 0% 20% 40% 60% 80% 100% Biological similarity 0% 20% 40% 60% 80% 100% Chemical similarity Epoxiconazole Fenbuconazole Tetraconazole Triadimenol Cyproconazole Difenoconazole Itraconazole Propiconazole Paclobutrazol Ametryn Prothioconazole PFOS Atrazine PFHxS PFOA PFHxA Melamine Cyanuric acid GenX 5-Methyl-1H-benzotriazole TFMSA 1H-1,2,4-Triazole PFBA PFBS PFPrA Deethylatrazine Cyromazine Benzoguanamine 1,2,3-Benzotriazole Step 1: Select one compound as target Step 2: Extract all in vitro activity data from ToxCast Step 3: Calculate similarity based on binary hitcall data* Chem similarity: Maccs/Tanimoto; Biological similarity: Tanimoto Biological similarity can be used to better distinguish compound classess Tebuconazole Data density Name total assays shared_active Biol_sim Chem_sim Tebuconazole 668 (100%) 82 100% 100% Cyproconazole 484 (72%) 21 45% 86% Paclobutrazol 483 (72%) 19 32% 84% Prothioconazole 214 (32%) 520% 73% Propiconazole 481 (72%) 28 35% 72% Tetraconazole 388 (58%) 30 48% 69% Epoxiconazole 204 (31%) 19 59% 69% Fenbuconazole 458 (69%) 24 48% 68% Triadimenol 495 (74%) 35 45% 67% Difenoconazole 322 (48%) 33 41% 66% triazole acetic acid 64% triazole alanine 60% Bitertanol 232 (35%) 11 69% 58% Itraconazole 43 (6%) 440% 57% Atrazine 538 (81%) 10 15% 43% 5-methylbenotriazole 313 (47%) 00% 43% 4-methylbenzotriazole 43% Benzotriazole 516 (77%) 00% 39% 1, 2, 4, triazole 414 (62%) 00% 39% Pyroxsulam 38% Deethylatrazine 208 (31%) 00% 36% Cyromazine 505 (76%) 00% 35% Ametryn 494 (74%) 12 22% 34% Ammeline 28% Benzoguanamine 334 (50%) 00% 27% Ammelide 27% Cyanuric acid 489 (73%) 00% 25% Melamine 55 (8%) 00% 23% GenX 287 (43%) 00% 13% PFOA 499 (75%) 46% 12% PFHxA 512 (77%) 35% 12% PFBA 272 (41%) 00% 12% PFPrA 266 (40%) 00% 12% TFA 12% PFOS 455 (68%) 17 16% 11% PFHxS 259 (39%) 111% 11% S2 3 ( %) 00% % - Data density needs to be considered -Atrazine for example has no ToxCast data -Relevance of in vitro data is also an open question in this existing dataset Ranking of compounds based on NAMs 29.10.2025 8 13 12 12 12 11 10 10 8 8 7 6 4 1111111 0 2 4 6 8 10 12 14 Propiconazole Tebuconazole Fenbuconazole Tetraconazole Ametryn Cyproconazole Difenoconazole Paclobutrazol Triadimenol Bitertanol Atrazine Deethylatrazine 4-… PFBS PFHxS PFOS PFHxA PFOA GenX Cyromazine Melamine Cyanuric acid Ammeline Ammelide Benzoguanamine 1, 2, 4, triazole Benzotriazole triazole acetic… 5-… triazole alanine Pyroxsulam TFMSA TFA PFPrA PFBA PFEtS Ranking of the compounds -Based on in vitro Activity Grouping of Compounds in ZeroPM -log units (µg/L) Azoles cluster together Triazines cluster together, mainly inactive PFAS cluster together Ranking according to „toxicity“ Prioritize candidate PMT/vPvM chemicals for additional data acquisition or towards identifying solutions aimed at preventing exposure