Naturally Occurring New Peptides From Velvet Antler of Cervus nippon Temminck: Structural Characterization and Organic Synthesis, Antioxidant and Anti-melanogenic Effects Using In Vitro Cell Models and In Vivo Zebrafish Models.

The aim of this study was to reveal the chemical basis and mechanism of antioxidant and anti-melanin effects of the Cervus nippon Temminck velvet antler. The water extract and four enzymatic products (Alcalase, pepsin, protamex, and trypsin) were screened for their activities by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 1,1-diphenyl-2-trinitrophenyl hydrazine free radical scavenging activities. Ultrafiltration and liquid chromatography-tandem mass spectrometry characterization followed. Molecular docking predicted the most active peptide (VVYPW, a known oligopeptide first identified in velvet antler), which was then synthesized using solid-phase peptide synthesis technology. The mechanism of antioxidant activity and antimelanotic activity of the synthesized peptide was tested in vitro (HaCaT and B16F10 cells) and in vivo (zebrafish model). The synthesized VVYPW peptide (yield: 67.3%; purity: 98.21%) modulated intracellular GSH-PX, SOD, and MDA activities, neutralized reactive oxygen species production, inhibited tyrosinase activity and melanin synthesis, and down-regulated tyrosinase, tyrosinase-related protein-1, tyrosinase-related protein-2 and microphthalmia-associated transcription factor messenger RNA expression and reduced zebrafish skin pigmentation. These results suggest that VVYPW has excellent antioxidant and anti-melanin effects properties and has the potential for the development of anti-aging and skin-whitening products.