Caffeine, although not a nucleoside analog, is known to inhibit the replication of herpes simplex virus-1 (HSV-1) and has been shown to significantly limit the spread of HSV infection in vitro. The therapeutic efficacy of caffeine was examined in a murine cutaneous infection model. The midflanks of 6-week-old BALB/c mice were infected with HSV cutaneously after application of 10% caffeine (Cafon) gel, and was reapplied to the midflank 5 times daily thereafter. Treatment with Cafon gel significantly retarded the development of skin lesions. Both midflanks were cutaneously infected, and a placebo and active gel were applied to the right and left midflanks respectively. Cafon gel significantly retarded the appearance of vesiculation and reduced the number of vesicles compared with the placebo gel. Cafon gel was as effective as 5% acyclovir ointment, and no significant difference was observed in the development of local lesions between these two topical preparations. The efficacy of Cafon gel also corresponded to that of oral treatment with 5 mg/kg or more of acyclovir in our cutaneous infection system. These results suggest that Cafon gel can be useful for the topical treatment of cutaneous HSV infection.