Hecogenin‑nitrogen mustard hybrids with improved anti-breast cancer activity: Design, synthesis, and biological evaluation

To enhance the efficacy of hecogenin (HCG) against breast Cancer cells, we designed and synthesized two series of new HCG‑nitrogen mustard hybrids (4a-4f and 5a-5f) by linking benzoic acid mustard or chlorambucil to HCG via amino acid residues. The derivatives were screened to assess their anti-proliferative activity against three human breast Cancer cell lines (MDA-MB-231, MDA-MB-468, and MCF-7), and one normal human breast MCF-10A cell line. Among the synthesized compounds, hybrid 5d exhibited the most potent anti-proliferative activity against the triple-negative breast Cancer cell line MDA-MB-231, with an IC₅₀ value of 2.2 μM. This represents a 27.2-fold increase in potency compared to the parent compound HCG (IC₅₀ = 59.8 μM). Furthermore, hybrid 5d exhibited low toxicity toward MCF-10A cells (IC₅₀ > 100 μM), indicating certain selectivity. Notably, the transwell migration assay revealed that hybrid 5d significantly inhibited the invasion of MDA-MB-231 cells. Preliminary mechanism studies indicated that hybrid 5d induced G2/M phase arrest and Apoptosis via the mitochondria-related apoptotic pathway, as well as caused DNA damage. Collectively, these results suggest that hybrid 5d is a promising lead compound for anti-breast Cancer research worthy of further investigation.

Amino acid; Anti-breast cancer; Hecogenin; Mitochondrial pathway; Nitrogen mustard.