Antidiarrheal Potential of Psoralidin Through Modulation of <scp>COX</scp> , μ‐Opioid, and L‐Type Calcium Channel Signaling Pathways: In Vivo and In Silico Investigations

ABSTRACT Background Psoralidin (PSN), a prenylated coumestrol, demonstrates a broad spectrum of pharmacological properties, such as anti‐inflammatory, anticancer, anti‐diabetic, and neuroprotective activities. This study aimed to evaluate the therapeutic potential of PSN as an antidiarrheal agent using in vivo experimentation in a castor oil‐induced chick model, supported by in silico receptor‐binding analyses. Methods Chicks were treated orally with PSN at doses of 5, 15, and 30 mg/kg and compared with standard reference drugs, including bismuth subsalicylate (BSS, 10 mg/kg), loperamide (LOP, 3 mg/kg), and nifedipine (NFN, 2.5 mg/kg). Diarrheal onset, stool frequency, and intestinal fluid secretion were recorded. Complementary molecular docking, pharmacokinetic, and toxicity assessments were performed to predict drug–receptor interactions and safety profiles. Results Among all treatment groups, PSN‐15 at its most effective dose produced the most significant reduction in parameters, lowering the diarrheal secretion (DRS) score to 138.00 ± 2.91 mg and the number of stool output (NSO) to 6.80 ± 1.09 events compared to the NC group. In silico analysis demonstrated that PSN formed stable binding complexes with high binding affinity (BA) for cyclooxygenase‐2 (COX‐2) (−9.2 kcal/mol), as well as for μ‐opioid receptor, COX‐1, and L‐type calcium channel. Computational toxicity profiling suggests a favorable preliminary safety profile for PSN. Conclusion In conclusion, PSN demonstrated significant antidiarrheal activity by reducing stool frequency, intestinal secretion, and delaying onset of diarrhea, likely through multi‐target modulation of COX, μ‐opioid, and L‐type calcium channel pathways. Nonetheless, future studies will focus on detailed pharmacokinetic profiling, long‐term toxicity assessment, and validation of PSN's antidiarrheal efficacy in advanced preclinical models to support its therapeutic potential.