Nowadays, peony seed oil (PSO) and chia seed (CS) provide a vast potential in the industries of animal feed, food, nutraceutical, pharmaceuticals, and health. In this paper, study on PSO and CS were added into biscuits, and improvement in the nutraceutical properties of biscuits. The studies indicated that biscuits of acceptable overall quality could be prepared using the content of incorporation of PSO (15 g) and CS (12 g) formulations. The total dietary fiber content was positively correlated with different content of PSO and CS. The body weight of the mice in high dose group was lower than that of control group and other groups, indicating that the components PSO and CS in the biscuits could significantly control the body weight of the mice. From the fourth week, average daily food intake of mice in high dose group was significantly lower compared to control group. However, the grasping power of mice in high dose group increased significantly. With the increase in the content of PSO and CS of biscuits, the DPPH radical scavenging activity increased, too. Thus, the results indicated that whole wheat biscuits supplemented with PSO and CS could be considered as dietary fiber-enriched food, and have antioxidant properties.