Intralesional (IL) treatment of cutaneous leishmaniasis with pentavalent antimonials has greatly reduced the systemic toxicity imposed by the conventional intramuscular and intravenous injections. Nevertheless, therapeutic failure with antimonials can occur due to drug resistance and/or short retention in the inflamed skin. Here we evaluated the safety and efficacy of IL treatment with injectable amphotericin B (AmB), a powerful antileishmanial drug that is less prone to drug resistance, but due to its amphiphilic nature is formulated with the ionic detergent sodium deoxycholate (Deox). Thus, Deox-AmB reconstituted with injectable water as directed was evaluated in vivo in L. amazonensis - infected mice in comparison with equivalent Deox or AmB alone. On days 7, 10, 14 and 17 of infection, the infected ears were injected with 10, 50 or 150 μg of AmB/dose. After 34 days of infection, AmB was most effective in killing the parasites at 150 μg/dose. However, at that effective dose Deox-AmB produced severe skin necrosis due to Deox. Our findings indicate that the necrotizing effect of Deox should be considered before using commercial Deox-AmB formulations containing this stabilizing agent for intralesional treatment.