Introduction: This study investigated the anti-asthmatic effects of Spondias pinnata using isolated guinea pig ileum and performed molecular docking analysis to identify potential asthma-related molecular targets. Methodology: The extract of S. pinnata underwent phytochemical analysis and isolated β-sitosterol, Epicatechin using column chromatography. The anti-asthmatic activity of the extract was assessed using isolated guinea pig ileum, measuring its inhibitory effect on histamine-induced contractions, mast cell degranulation inhibition, and acute toxicity studies were conducted. The extract's impact on bronchospasm inhibition was measured, including its effect on pre-convulsive time. Molecular docking analysis assessed interactions between β-sitosterol, Epicatechin, IL-13, and endothelial nitric oxide synthase.