Abstract: Using wheat peptide as the raw material, the wheat peptide sports beverage was developed. The optimum formula of the beverage was researched by single-factor and response surface test. The climbing pole time, content of lactic acid (LA), blood urea nitrogen (BUN) and malondialdehyde (MDA), activity of superoxide dismutase (SOD) and glutathione peroxide dismutase (GSH-Px) were respectively measured in different group mice after exercise, to explore anti-exercise fatigue and antioxidant of this beverage in vivo. The results showed that the optimal formula of this beverage was the combination of 3.5% of wheat peptide, 4.2% of erythritol, 0.2% of citric acid, and 0.35% of compound stabilizer. Under predicted condition, sensory score of the beverage reached 95.7 scores. The beverage presented light yellow, possessed peculiar fragrance of wheat peptide, proper sweet and sour taste, unique flavor, and without obvious precipitate, and content of glutamine reached 0.69%±0.02%. Compared with control group, the wheat peptide sports beverage at different dosage could prolonged climbing pole time on mice (P<0.05, P<0.01), increased activity of SOD and GSH-Px that were endogenous antioxidant enzymes (P<0.05, P<0.01), meanwhile decreased generated content of MDA, LA and BUN (P<0.05, P<0.01) after exercise. Therefore, the wheat peptide sports beverage had better activity anti-exercise fatigue and antioxidant in vivo.