A trimer of cholic acid was prepared via a simple isocyanate linkage. Fluorescence and UV-vis spectroscopic studies showed that the same cholane trimer can form either a hydrophobic cavity in aqueous solutions or a hydrophilic cavity in an apolar solvent using pyrene and a derivative of pyrene as probes. The dynamic inversion of the cavities was demonstrated for the first time by a simple change of the polarity of the solvent media by adjusting from an aqueous buffer to tetrahydrofuran and vice versa. The NMR study of a dimer model compound of cholic acid indicated a restricted conformation exist in the solvent mixture of chloroform and methanol.