Abstract

While artemisinin and its derivatives demonstrate broad antitumor potential, the stereochemical influence on the bioactivity of higher-order artemisinin assemblies remains inadequately characterized. Herein, we report the synthesis, chromatographic separation, and structural elucidation of four stereoisomeric artemisinin trimers, followed by systematic evaluation of their antitumor efficacy against MCF-7 and MDA-MB-231 breast cancer cell lines. All trimers exhibited potent cytotoxicity against MCF-7 cells (IC50< 0.09 μM), with trimer6a(β,β,β) demonstrating robust antitumor activity in both in vitro and in vivo xenograft models. Remarkably, pronounced stereochemistry-dependent activity emerged against MDA-MB-231 cells:6adisplayed approximately 100-fold greater potency than6b(β,β,α) and 6.6-fold superiority over gemcitabine. Mechanistic investigations revealed that6adownregulates Cyclin D1, CDK4, and CDK6 expression, thereby inducing G0/G1 phase cell cycle arrest. These findings underscore the pivotal role of stereochemical configuration in modulating artemisinin trimer bioactivity and provide rational guidance for structure-based design of artemisinin-derived anticancer therapeutics.