<p>Leishmaniasis is a public health problem caused by protozoan of the genus Leishmania. This parasitic vector-borne disease, presents different clinical manifestations including cutaneous, mucocutaneous and visceral leishmaniasis. The lack of an effective and nontoxic antiparasitic treatment highlights the need to develop new leishmanicidal agents. To address this challenge, the use of bioinformatics and molecular modelling will help on the identification of specific biological targets and to determine the structural and molecular properties that confer biological activity to specific compounds. On the other hand, biological evaluation is a critical aspect in the discovery of new chemical structures with chemotherapeutic properties, and flow cytometry has emerged as a potent tool for the realization of biological assays. The conjunction of in silico and in vitro studies provide valuable information to direct the selection of molecules for in vivo experiments, reducing costs, time and the excessive use of experimental animals. This chapter aims to discuss theoretical techniques and in vitro evaluation tools available for a rational design of new, effective and low cost leishmanicidal agents.</p
