Assessment of Anti‐Diarrheal Effect of Schaftoside Through μ‐Opioid and Cyclooxygenase Interaction Pathway: In Vivo and In Silico Studies

BACKGROUND: Diarrhea remains a significant global health burden, necessitating the discovery of safer and more effective therapeutic agents. Schaftoside (SCF), a bioactive flavonoid, has demonstrated diverse pharmacological properties, though its anti-diarrheal potential remains underexplored. This study aimed to evaluate the anti-diarrheal activity of SCF and elucidate its underlying mechanisms. METHODS: The in vivo efficacy of SCF was assessed in a castor oil-induced diarrhea model using 2-day-old chicks. Animals were pretreated orally with SCF (5, 10, 20 mg/kg), loperamide (3 mg/kg), or bismuth subsalicylate (10 mg/kg). Diarrheal parameters, including latency period, stool frequency, and secretion weight, were recorded. Complementary in silico molecular docking was performed to investigate SCF's binding affinity (BA) and interactions with the μ-opioid receptor (PDB ID: 8EFB), cyclooxygenase-1 (COX-1; 6Y3L), and cyclooxygenase-2 (COX-2; 5F19). RESULTS: SCF administration produced a significant, dose-dependent anti-diarrheal effect. The highest dose (20 mg/kg) markedly reduced diarrheal secretion and stool frequency while prolonging the onset of diarrhea, with efficacy comparable to standard drugs. In silico analysis revealed strong binding affinities of SCF for the μ-opioid receptor (-9.8 kcal/mol) and COX-2 (-9.0 kcal/mol), supported by multiple hydrogen bond and hydrophobic interactions with key active-site residues. CONCLUSION: These findings demonstrate that SCF possesses substantial anti-diarrheal activity, potentially mediated through dual modulation of the μ-opioid receptor and COX-2 pathways. SCF represents a promising natural candidate for further development as a therapeutic agent for diarrhea management.