Owing to their unique properties, ferrocene compounds are gaining increasing interest for biological applications, particularly as enzyme inhibitors. Phosphonate derivatives including a ferrocenyl moiety were synthesized by reaction of dimethyl- and diphenylphosphite with ferrocenyl methyl maleimide. The ferrocenyl diphenyl phosphonate complex was characterized by X-ray diffraction. The ability of these organometallic compounds to inhibit the enzymatic activity of the serine esterases acetyl- and butyrylcholinesterase was investigated. It appeared that the new ferrocenyl phosphonates inhibited both enzymes by competitive, mixed or non competitve mechanisms with inhibition constants in the range 35–1000 µM. Both compounds also behave as slow time-dependent inactivators of butyrylcholinesterase. The presence of the ferrocenyl entity seems then to have a dramatic effect on the biochemical behavior of the systems.