Ov-16 [4-(3,4-dihydroxybenzoyloxymethyl)phenyl-O-β-D-glucopyranoside] inhibits melanin synthesis by regulating expressions of melanogenesis-regulated gene and protein.

Ov-16 (4-(3,4-dihydroxybenzoyloxymethyl)phenyl-O-β-D-glucopyranoside), a polyphenolic glycoside that is isolated from oregano (Origanum vulgare L.), can scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals. This investigation is the first to study in detail the hypopigmentary properties of Ov-16. It demonstrates that 0-1000 μg/ml Ov-16 inhibits the activity of mushroom tyrosinase (Tyr) in a concentration-dependent manner. The inhibitionary Tyr kinetics of Ov-16 towards the oxidation of L-DOPA was found to be uncompetitive. Following the treatment of human skin premalignant kerationcyte HaCaT cells, human skin fibroblast Hs68 cells and mice melanoma B16 cells with Ov-16 (0-100 μg/ml), cell viability was >98%, suggesting that Ov-16 is non-toxic. Ov-16 can reduce cellular Tyr activity, DOPA oxidase activity and melanin synthesis in B16 cells that are stimulated by the α-melanocyte-stimulating hormone (α-MSH). Moreover, Ov-16 inhibited the production of melanin in Streptomyces bikiniensis without affecting the growth of the microorganism. The treatment of B16 cells with Ov-16 considerably reduced the gene expressions of melanocortin-1 receptor (Mc1r), microphthalmia-associated transcription factor (Mitf), Tyr, tyrosinase-related proteins-2 (Trp-2) and Trp-1, as determined by RT-PCR. The expressions of Mc1r, Mift, Tyr, Trp-2 and TrpP-1 protein in Ov-16-treated B16 cells were also significantly reduced, as determined by western blotting and fluorescent staining analysis. These results suggest that Ov-16 exhibits hypopigmentary performance.