Real-world results from a Machine Learning-guided, phenotypic High-Throughput Screen for novel antibiotics

Antimicrobial resistance is an urgent global health threat, with over 2.8 million multidrug-resistant infections killing over 35,000 annually in the US. Machine Learning (ML) has emerged as a potential solution to improve efficiency of antibiotic high-throughput screens (HTS). We report ML-guided high-throughput screening against E. coli . Large-scale Learning-to-Rank models were trained on public and proprietary datasets to maximize phenotypic inhibition and minimize human cell cytotoxicity. We evaluated several pre-plated compound libraries and a set of “cherry-picked”, structurally novel compounds. We screened against a hyperpermeable lptD ^- mutant, followed by hit confirmation, profiling, cytotoxicity counter-screening, and MOA determination. Results demonstrated a doubled hit rate and 3X fewer toxic hits. Additionally, activity improved against both Wild Type E. coli and the lptD ^- mutant. ML models showed robust predictive power on structurally dissimilar compounds. The combination of large-scale HTS, ML innovation, and both library-wise selection and cherry-picking strategies distinguishes this study in the antibiotic discovery field.