Entadamide A-CO-C12 is a derivative of Entadamide A, demonstrating potent anticancer activity, particularly effective against breast cancer cell lines. This compound promotes apoptosis, inhibits cellular migration, spheroid formation, and stem cell-like populations. Entadamide A-CO-C12 also suppresses tumor growth in 4T1 mouse models and is utilized in breast cancer research.

Entadamide A-CO-C12 (compound 3u) (48 hours) demonstrates antiproliferative effects against various human cancer cell lines (MDA MB-231, DU-145, MCF-7, 4T1, NCI-H460, HepG2, and HEK-293) with IC50 values of 19.11, 3.04, 2.82, 3.26, 20.98, 18.89, and 77.69 μM, showing selective anticancer activity against breast cancer cells (MCF-7 and 4T1). At concentrations of 1.5-3 μM for 48 hours to 5 days, it inhibits MCF-7 cell migration and downregulates EMT-related transcription factors. Additionally, Entadamide A-CO-C12 (1.5-3 μM; 7 days) impedes spheroid formation and reduces cancer stem cell characteristics in MCF-7 cells. It promotes apoptosis in MCF-7 cells (1.5-3 μM; 24-48 hours) by balancing the transcriptional and translational regulation of anti-apoptotic (BCL-2 and survivin) and pro-apoptotic (BAX) factors, triggering mitochondrial depolarization and activation of intrinsic apoptotic pathways. Furthermore, Entadamide A-CO-C12 (1.5-3 μM; 8-24 hours) induces a significant ROS increase in MCF-7 cells in a dose-dependent mannerand, at 1.5-3 μM for 48 hours, reduces CD90 and CD133 levels, induces DNA damage, and triggers apoptotic nuclear condensation in MCF-7 cells dose-dependently. In addition, Entadamide A-CO-C12 (1.5-3 μM; 48 hours) enhances the sensitivity of MCF-7 cells to Doxorubicin by modulating ABCG2-mediated efflux pathways. It interacts partially with ABCB1/ABCG2 and BMI1, aiding in the regulation of drug resistance and stemness.

Entadamide A-CO-C12, administered intraperitoneally at 30 mg/kg once daily for 15 days starting on day 7, demonstrates potent in vivo anticancer activity without systemic toxicity in a 4T1 orthotopic breast cancer mouse model.