Identification of hyaluronidase inhibitory peptides from the hydrolysates of royal jelly proteins for skin anti-aging applications

Skin aging is characterized by reduced hydration, elasticity loss, wrinkles, and impaired skin function. Hyaluronidase-mediated hyaluronic acid degradation, together with oxidative stress, inflammation, and fibroblast senescence, is a key driver of this process. Therefore, safe hyaluronidase inhibitors with additional bioactivities may offer potential for skin-aging intervention. In this study, royal jelly protein hydrolysates (RJPH) were generated by enzymatic hydrolysis and characterized using UHPLC-MS/MS-based peptidomics. Potential hyaluronidase-inhibitory peptides were screened through virtual screening, molecular docking, and in vitro enzyme inhibition assays. A novel hyaluronidase-inhibitory peptide, with the amino acid sequence abbreviated as LVSAFK, was further evaluated using molecular dynamics simulation, the oxygen radical absorbance capacity (ORAC) assay, and cell-based assays in human skin fibroblastsusing lipopolysaccharide (LPS)-induced inflammation and H 2 O 2 -induced senescence models. LVSAFK displayed hyaluronidase inhibitory activity with an IC50 of approximately 301.6 μM. Docking and molecular dynamics analyses indicated stable binding mediated by hydrogen bonds, salt bridges, and cation-π interactions. In addition, LVSAFK exhibited peroxyl radical-scavenging activity, suppressed LPS-induced IL-1 β production, and reduced SA- β -galactosidase-positive senescent fibroblasts under H 2 O 2 stress. Collectively, these findings identify LVSAFK as a multifunctional royal jelly-derived peptide with in vitro activities relevant to skin aging, warranting further mechanistic and in vivo studies for potential anti-skin-aging applications.