Background: Considering the difficulties, adverse effects and unsuitable response to commonly used drugs; it is essential to find an alternative, particularly local treatment for cutaneous leishmaniasis (CL). Objective: Determination of efficacy of topical paromomycin (Paromo-U ointment) against CL caused by L.major in mouse model. Materials and Methods: Skin lesions were created 1 to 3 months after inoculation of L.major promastigotes to the base of tail of small, white mice (Outbreed). Then the mice were randomly divided to three groups including interventional (Treatment by Paromo-U), control 1 (Treatment by urea) and control 2 (Treatment by distilled water) groups. The mice were treated topically twice a day for 8 weeks. Results: The mean lesion diameter of the lesions prior to treatment in interventional and control groups (Urea and distilled water) were measured 10.9, 5.9 and 6.0 mm respectively and changed to 4.0, 12.7 and 14.3 mm 8 weeks after treatment. No Leishman bodies were observed within the lesions of interventional group, whereas they were seen in all control groups. Conclusion: Paromo-U ointment was effective in the treatment CL caused by L.major in mouse model.