This study evaluated the sub-chronic oral toxicity of freeze-dried deer velvet antler (DVA) extract in ICR mice over a 90-day period. The extract was administered orally to male and female mice at doses of 1,000, 2,000, and 5,000 mg/kg body weight daily; the control group (0 mg/kg body weight) received vehicle only. No clinical signs of toxicity or mortality were observed. Body weight gain, food consumption, and general behavior remained normal across all groups. Hematological analysis revealed no dose-dependent alterations in hemoglobin levels, platelet characteristics, or red or white blood cell counts. Males administered 2,000 mg/kg DVA exhibited a slight increase in reticulocyte count, whereas females exhibited reduced count at 5,000 mg/kg; however, neither finding was doseresponsive or toxicologically significant. Serum biochemistry including liver and kidney function markers (ALT, AST, -GT, BUN, and creatinine) remained within normal range, and no evidence of organ damage was observed. Notably, male mice exhibited a significant reduction in serum triglycerides compared with those in the control group, suggesting potential lipid-modulating effects of DVA; however, this finding requires further investigation. Based on these findings, the no-observed-adverse-effect level (NOAEL) of DVA extract was determined to be greater than 5,000 mg/kg/day in both sexes. These results support the safety profile of DVA extract at the tested doses, and its potential metabolic benefits.