A Leishmania donovani promastigote line resistant to 160microM sitamaquine, named SITA-160R, was selected in vitro by continuous stepwise drug pressure. SITA-160R line was able to infect murine peritoneal macrophages in vitro in the same manner as the wild-type line (WT) but was less infective for Balb/c mice than its parent WT clone. This line was about five and three times more resistant to sitamaquine than the WT line on promastigote and intramacrophage amastigote forms, respectively. No cross-resistance with other antileishmanial agents was observed and this resistance was stable when parasites were subcultured in drug-free medium for a long time or after in vivo passage.