Abstract: OBJECTIVE To design and synthesis of ferrocenyl heterocyclic derivatives and investigate anti-triple negative breast cancer activity. METHODS A series of ferrocenyl derivatives were designed and synthesized from ferrocenyl chalcone, and their anti-breast cancer activities were evaluated by CCK8 assay. RESULTS Twenty-eight ferrocenyl heterocyclic derivatives were synthesized and the structures had been confirmed by ¹H-NMR and MS spectra. The preliminary biological results showed that all synthesized ferrocenyl derivatives showed selective anticancer activity that were more potent against MDA-MB-231 cells than MCF-7, which also showed moderate inhibitory activity, against MDA-MB-231 cell lines, and imidazole heterocyclic compounds had more potent anti-tumor than corresponding pyrazole and pyrimidine derivatives, specifically, compound 28aIC₅₀=(1.6±0.23)μmol·L^-1 showed about 6 and 10-fold potency than lead compound 3IC₅₀=(10.7±1.41)μmol·L^-1 and tamoxifenIC₅₀=(13.7±1.17)μmol·L^-1, against MDA-MB-231 cell lines, and these ferrocenyl derivatives were not toxic to normal cells. CONCLUSION This study provides information and basis for development of ferrocenyl derivatives with anti-triple negative breast cancer activity.