Abstract:

Currently, there is little research on tetrapeptides with melanin inhibitory activity. This article aims to study melanin inhibition by constructing a virtual peptide library and establishing a design and screening method to select and evaluate tetrapeptides with melanin inhibitory activity. Based on a virtual peptide library, technologies such as molecular docking, activity prediction, and force analysis were used to screen the target active tetrapeptides, and then solid-phase peptide synthesis was employed for directed synthesis of the target tetrapeptides. The efficacy of the target active tetrapeptides was further evaluated from biochemical detection, cell models, and molecular transcription perspectives. Four tetrapeptides are selected: Peptide-1 （CHWH）, Peptide-2 （RVGW）, Peptide-3 （LHSY）, and Peptide-4 （LFSY）. Compared to the other three tetrapeptides, Peptide-1 exhibits better Cu²⁺ chelation, ABTS positive ion scavenging, and tyrosinase inhibitory activity. The IC₅₀ values for tyrosinase activity and melanin production inhibition in B16 cell models are 0.28 and 0.20 g/L, respectively, and it dose not significantly affect cell proliferation below a mass concentration of 10 g/L. Transcriptional studies also further show that Peptide-1 can decrease the intracellular levels of MITF （microphthalmia-associated transcription factor） and TYR （tyrosinase） gene expression in a concentration ration-dependent manner.

Key words: active tetrapeptide, B16 cells, inhibition of tyrosinase activity, melanin production