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Background: Ferrocene is a potential pharmacophore for drug design and drug discovery. Methods: Based on our previous good achievements (Med. Chem. commun., 2014,7,968-972), nineteen novel structures of 1,1'-ferrocene diformates bearing isoxazole moiety (3a-3s) were firstly synthesized in the current work and characterized by 1H NMR, 13C NMR, ESI-MS. Then, their cytotoxicity to A549, HCT116 and MCF-7 cell lines was evaluated using the MTT method. Results: The results showed that most compounds exhibited higher potent cytotoxicity to A549, HCT116 and MCF-7 cell lines. Conclusion: Especially, 3b, 3h, 3k, 3l, 3m, 3n, 3o, 3p and 3s simultaneously exhibited stronger inhibitory activity towards A549, HCT116 and MCF-7 cell lines than that of the reference drug cisplatin, which can be regarded as very promising metal-based lead compounds for anticancer agents.