Effects of calcium and lipophilicity on transport of clodronate and its esters through Caco-2 cells.

Clodronate, like other bisphosphonates, is poorly absorbed from the gastrointestinal tract, mainly due to its high hydrophilicity and ability to form complexes with divalent cations in the gastrointestinal tract. One strategy for improving oral absorption of these types of molecules is to develop more lipophilic derivatives. The importance of lipophilicity and calcium chelation in the absorption of clodronate was evaluated by studying the penetration of clodronate and its mono-, di-, and triphenyl esters through human intestinal Caco-2 cells. The transport rates of [(14)C]-clodronate and its mono-, di-, and triphenyl esters were quantified by calculating their apparent permeability coefficients (P(app)) both in normal (1.3 mM) calcium concentration and in 'minimum-calcium model'. The transport rate of 1 mM clodronate was very low (0.25 x 10(-7) cm/s), while the removal of calcium from the apical side increased this transport rate 6-fold. The transport rate of clodronate was increased with increasing dose. Mono- and diphenyl esters did not significantly enhance the transport of clodronate. Triphenyl ester, however, increased the transport rate 17-fold compared with parent clodronate. Removal of calcium did not affect the transport rates of di- or triphenyl esters, which indicated that the esterification of hydroxyl groups of clodronate decreased calcium complex formation. These results indicate that clodronate is transported paracellularly through Caco-2 cells and that calcium decreases strongly its absorption. They further suggest that at least three phosphate hydroxyl groups need to be substituted until the permeation route is changed from paracellular to transcellular.