Melanin is a group of natural pigments and is the major factor responsible for skin and hair color in mammals. It has several beneficial functions, including skin protection from harmful ultraviolet (UV) radiation and prevention of cancer development. However, abnormal accumulation and hyperpigmentation of melanin result in pigmentation disorders such as freckles, melasma, chloasma, ephelides, senile lentigines and melanoderma and can induce inflammation such as eczema, allergic contact dermatitis, and irritant contact dermatitis, which can lead to serious and emotionally distressing problems. We previously demonstrated that the chalcone derivative 1-(2-cyclohexylmethoxy-6-hydroxy-phenyl)-3-(4-hydroxymethyl-phenyl)-propenone (chalcone-21) exhibited anti-melanogenic activity in B16F10 melanoma cells. To increase its anti-melanogenic activity, we synthesized 21-kinds of novel compounds as an its derivative and determined their inhibition effects of melanin production in a-MSH-induced B16F10 cells. Among them, chalcone-21-21 exhibited the strongest inhibition of cellular melanin production with an IC50 value of 0.54 mM, compared with 4.9, 38.5 and 148.4 mM of IC50 values of chalcone 21, kojic acid and arbutin, respectively. As the molecular mechanisms, we identified chalcone 21-21 suppressed the expression and transcriptional activity of tyrosinase, the rate-limiting enzyme of melanogenesis, as well as its enzyme activity. Chalcone 21-21 suppressed also the expression of TRP-1, TRP-2, and MITF. In addition, chalcone 21-21 suppressed not only phosphorylation of CREB and ERK1/2, but also the transcriptional activity of MITF and CRE. Our results demonstrated chalcone-21-21 is an effective anti-melanogenic agent and may benefit the treatment of hyperpigmentation disorders.