To investigate the protective performance of prestressed concrete structures against external impact and the influence of prestress level on the impact resistance of the structure, impact tests were conducted on reinforced concrete target plates under non-prestressed, low-prestressed, and high-prestressed conditions. The effects of different prestress levels on the impact resistance of reinforced concrete target plates were analyzed. Additionally, numerical simulations were performed for the impact tests of reinforced concrete target plates with varying prestress levels, and the simulation results were in good agreement with the experimental results. The research results show that, in comparison with the ordinary reinforced concrete target plates, prestress can enhance the local anti-penetration capacity of the target plates to a certain degree, reducing the penetration depth as well as the cratering range. On the other hand, the application of prestress increases the overall stiffness of the reinforced concrete target plate, once the target plate is penetrated to a certain depth, the spalling area is larger compared to that of non-prestress or low-prestress reinforced concrete target plates. Hence, the effects of rear fragmentation need to be taken into account.