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Trypanosomatids diseases are well known for a long time. These blood-borne parasites peculiarly present vector and host alternating forms with well-differentiated biochemistry, morphology and ways to escape. Recent studies about the metabolic ways used by these parasites are of great scientific interest, especially the divergence of parasite and human host metabolic pathways. Thus, the obtaining of energy (glycolytic pathway), polyamine biosynthesis, sterol biosynthesis, microtubule biosynthesis, folate metabolism, and the DNA topoisomerase, trypanothione redutase, hypoxanthine-guanine phosphoribosyltransferase, nitric oxide synthase, arginase, cysteine protease, and superoxide dismutase enzymes have proved to be important targets to new anti-trypanosomal agents.