Background: The azole anti-mycotic agents, such as ketoconazole and sertaconazole, have been shown to exhibit anti-inflammatory activity and anti-cancer effect via p38 signaling pathway. p38 pathway has been known to be related to melanogenesis. Objectives: We hypothesized that azole antifungal agents might affect the skin pigmentation. We herein investigated the effect of clotrimazole, one of azole antifungal agents, on melanogenesis. Methods: The toxic effects of clotrimazole on Mel-ab melanoma cells were assessed. To understand the mechanism of the effect of clotrimazole on melanogenesis in Mel-ab cells, melanin content and tyrosinase activity were measured. We further investigated the expression and degradation of tyrosinase and related signal transduction pathways. Results: We showed that clotrimazole led to reduced melanin content in melanoma cells. Clotrimazole led to decrease in tyrosinase protein level, without altering mRNA expression. Treatment of MG132, a proteasomal inhibitor, abolished both the suppression of melanin synthesis and the downregulationof tyrosinase level by clotrimazole. Furthermore, we demonstrated that the anti-melanogenic effect of clotrimazole is independent of p38 pathway and is related to extracellular signal-regulated kinase (ERK) and Akt signaling pathways. Conclusion: Clotrimazole inhibits melanin synthesis via proteasomal degradation of tyrosinase, and its anti-melanogenic effect is mediated through activation of ERK and Akt signaling pathways.