Anti-Melanogenic Activity of <i>p</i>-Chlorophenyl Benzyl Ether in α-MSH-Induced Mouse Melanoma B16F10 Cells

Abstract

Melanin is cutaneous pigment which level of its production determines skin complexion. Overproduction of melanin, frequently promoted by UV rays, results in darkening of the skin. Inhibition of tyrosinase activity, a core component in melanin biosynthesis, is one of the mechanisms of depigmenting agents. Hydroquinone and kojic acid are the examples of well-known whitening agents widely used in both pharmaceutical and cosmetic products. However, their adverse effect issues still needed to be overcome. A recent study showed that p-chlorophenyl benzyl ether (Cl-benz), a new synthetic compound, more strongly inhibited mushroom tyrosinase than kojic acid. In the current study, cytotoxicity, anti-melanogenic activity and anti-tyrosinase activity of Cl-benz were performed in mouse B16F10 melanoma cells compared to kojic acid. After 24 h of treatment on B16F10 cells, the cytotoxicity was not observed with Cl-benz and kojic acid. However, after incubation for 48 h, kojic acid at a concentration of 500 μM reduced cell viability less than 50%, whereas Cl-benz-treated cells showed negligible cytotoxicity. For cell-based assay, Cl-benz exhibited inhibitory effect similar to kojic acid. Melanin production in B16F10 cells was suppressed by Cl-benz in a dose dependent manner. One hundred micrograms of Cl-benz decreased melanin content in α-MSH by 66%. Moreover, the percentage of cellular tyrosinase activity of Cl-benz showed positive association with its corresponding melanin content. These results revealed that Cl-benz could inhibit melanogenesis via the mechanism of cellular tyrosinase inhibition. Accordingly, Cl-benz has potential to become a novel skin whitening agent in terms of efficacy and safety.

Synthesis of Fluorinated <i>N</i>- Benzylaniline Derivatives and Evaluations on Anti-Tyrosinase and Anti-Melanogenic Activities

“…The melanin measurement was determined as described previously with some modifications. [10] In brief, B16F10 mouse melanoma cells were cultured in 12-well plates at a density of 1 × 10 5 cells/well and incubated with 100 nM α-MSH and test samples or kojic acid (as a positive control) at different concentrations for 72 h. Subsequently, cells were detached with trypsin and transferred to Eppendorf tube then centrifuged at 3,000×g for 10 minutes. The supernatant was discarded then 10% DMSO/1 N NaOH was added into the Eppendorf tubes and heated at 80 ºC for 1 h to extract the melanin.…”