Multiclass Machine Learning-Based Discovery of Novel Scaffold Inhibitors Targeting ALK

Background: Anaplastic Lymphoma Kinase (ALK) is an oncogenic receptor tyrosine kinase implicated in several cancers. Despite the clinical success of ALK inhibitors, acquired resistance continues to drive the search for novel chemotypes. We developed a multiclass machine learning framework to classify ALK inhibitory activity using a curated ChEMBL dataset. Methods: Models were built using 2D molecular descriptors together with MACCS and ECFP4 fingerprints. Three widely used algorithms, Support Vector Machine (SVM), Random Forest (RF), and XGBoost, were applied for model development. Results: RF and XGBoost models demonstrated the best performance, achieving accuracies of ~0.75–0.79 with consistently high ROC–AUC values, particularly for fingerprint-based features. Bemis–Murcko scaffold analysis identified enriched chemotypes and underexplored scaffolds for further prioritization. The validated models were subsequently used to screen the Maybridge library, and compounds predicted to possess potential ALK inhibitory activity were prioritized for further computational evaluation. Applicability-domain filtering confirmed that the selected compounds occupied the predicted ALK inhibitor chemical space across multiple activity classes. The shortlisted compounds were subsequently evaluated by molecular docking to characterize their binding modes and interactions. Three candidate hits (SCR00078, SCR00073, and AW01085) were selected for further evaluation using 500 ns molecular dynamics simulations alongside the reference inhibitor Brigatinib. Simulation analyses revealed stable protein–ligand complexes and reduced conformational fluctuations relative to apo ALK, while MM/PBSA calculations identified SCR00078 and AW01085 as the most favorable binders. Conclusions: This integrated ML-to-simulation workflow prioritizes structurally novel candidate hits with predicted ALK inhibitory activity and provides an effective strategy for scaffold discovery and hit prioritization.