Calceolarioside B alleviates airway barrier dysfunction and inflammation via targeting P2Y6R in OVA-triggered asthma.

Asthma is a chronic inflammatory airway disease characterized by barrier dysfunction and excessive inflammatory response. In this study, we identified that a traditional Chinese medicine Akebia quinate extract alleviated asthmatic phenotype in vivo. Transcriptomic analysis revealed that Akebia quinate extract associated with inflammatory response and airway epithelial barrier remodeling. Using network pharmacology screened the top four active ingredients from Akebia quinate and further evaluated their effects on inflammatory cytokines and airway barrier indicators. Among them, compound Calceolarioside B exhibited the most potent anti-inflammatory and barrier-protective activities. Pharmacodynamic validation confirmed that Calceolarioside B alone significantly ameliorated asthmatic features. The candidate target of Calceolarioside B was identified as P2Y6R using AI-assisted deep learning, virtual target fishing, and molecular dynamics simulations. In vitro binding assays validated the interaction between Calceolarioside B and P2Y6R. Furthermore, Calceolarioside B treatment failed to produce additional therapeutic effects in the P2Y6R knockout asthmatic mouse model, confirming that P2Y6R was the functionally relevant target of Calceolarioside B. Subsequent molecular docking and point mutation experiments identified LYS-25 as the effective binding site for Calceolarioside B and P2Y6R. Collectively, these findings demonstrate that Calceolarioside B alleviates asthma by targeting P2Y6R, providing a mechanistic basis for its therapeutic potential and a rationale for developing target-specific interventions.