Endometrial cancer is one of the most common gynecological cancers, with global new cases, approximately 420,000 new cases and 98,000 global deaths annually. Emerging evidence suggests that the prostaglandin E2 receptor (EP2) plays a critical role in tumor progression, angiogenesis, and immune evasion. Withanolides, a class of naturally occurring compounds, possess anticancer activity; however, effect on EP2 receptor in endometrial cancer remains largely unexplored. This study aims to explore the interaction between Withanolides and EP2 receptor using molecular docking techniques, with PF-04418948 as the reference antagonist. A select number of these ligands, with anti-cancer activity, were evaluated for their ADMET and toxicity properties, and those with favorable drug-like properties, low toxicity profile, and no more than 1 Lipinski’s rule violation, were docked to EP2 (PDB ID: 7CX2). Molecular docking studies revealed three ligands, (Pubchem 161671, 265237, and 21679027) with significantly higher binding affinity scores compared to that of the reference compound. Pubchem 161671, showed the highest binding affinity at -16.6 kcal/mol. Post-dock analysis revealed interactions with key amino acids, VAL89, LEU298, SER305, and MET31, which are essential for the antagonist activity of the EP2 receptor enzyme (Yano et al., 2017). Significant interaction with critical amino acid residues suggested potential inhibition of EP2 receptor activity, offering a potential therapeutic approach for treating endometrial cancer. Overall, this study profers a deeper understanding of the potential of Withanolides as leads for EP2 targeted therapy in endometrial cancer.