Opening the black box: study of the multi-target anticancer mechanisms of Ecballium elaterium cucurbitacins via in vitro, network pharmacology, and molecular docking analyses.

Ecballium elaterium is a perennial plant species that belongs to the Cucurbitaceae family. In this study, we examined the biological activities and analyzed the chemical profiles of ethyl acetate, acetone, aqueous acetone, and water extracts from different parts (aerial parts, fruit peels, and seeds) of E. elaterium. We performed chemical characterization using UPLC-ESI-MS/MS. To investigate the biological activities of the extracts, antioxidant capacity, enzyme inhibition, and cytotoxicity assays were conducted. Thirty-eight compounds were detected in the extracts, and cucurbitacins and their glycosides were the major components in the aerial parts and fruit peels. The aqueous acetone extract of the seeds exhibited the highest ABTS radical scavenging activity (155.82 mg TE per g), whereas the acetone extract of the aerial parts showed the greatest reducing power. The ethyl acetate extract of the aerial parts exhibited the highest inhibitory activity against AChE and BChE. Overall, the seed extracts showed glucosidase-inhibitory activity. The ethyl acetate extract of the aerial parts exhibited notable cytotoxic activity against the HCT-116 cell line (IC50 of 90.44 µg mL-1). Network pharmacology analysis linked the cucurbitacin-rich constituents to a converging set of cancer-related protein targets, and molecular docking predicted favorable binding affinities toward key hub proteins such as EGFR and STAT3, offering a preliminary computational rationale for the observed antitumor effects. As a result, the different E. elaterium extracts may serve as potential sources of bioactive compounds for further study and development in health-promoting applications.