Abstract: Objective: To better develop high-value products from deer antlers, this study determined the optimal process conditions for the enzymatic digestion of deer antler peptides by alkaline proteases and evaluated their in vitro anti-blue light activity. Methods: Select four factors: digestion temperature, substrate ratio, digestion time and pH, three levels of each factor, based on unifactor experiment optimization, the yield of the enzyme digestion of the deer antler peptide was obtained, and the content of the deer antler peptide was determined. The survival rate of the Human immortal keratinocyte line (HaCat) cultured with different concentrations of deer antler peptide was measured by the CCK-8 method. The method of ELISA was used to analyze the secretion levels of Superoxide dismutase (SOD) and reduced Glutathione-reduced (GSH) secretion levels. The UV method calculated the ability of antler peptides to scavenge DPPH free radicals and OH free radicals. Results: The optimum conditions for the enzymatic digestion of antler peptides were determined to be 45 °C, 5 h, pH8.5, and a 6% enzyme-substrate ratio. The extraction rate of antler peptide was 52.7% at this time. With different concentrations (12.5, 25, 50, 75, 100 μg/mL), the proliferation activity of HaCat cells was reduced, and the cell survival rate was increased, significant difference (P<0.01). Meanwhile, the level of SOD, GSH secretion increased and the MDA content decreased (P<0.05). In addition, the scavenging ability of antler peptide on DPPH free radicals and OH free radicals was higher, with IC₅₀ of 0.98 and 0.63 mg/mL, respectively. Conclusion: At the same time, this study had verified that deer antler peptides have better in vitro anti-blue light activity. The results proved that antler peptides have certain effects on skin whitening, anti-aging and damage repair. These conclusions provided a theoretical basis for deer antler's deep processing, development, and utilization.