On basis of our previous research of the enteric-coated pellets loaded with aspirin or esomeprazole magnesium, the co-loaded enteric-coated pellets were prepared in this paper. Firstly, the aspirin pellet cores were fabricated by extrusion-spheronization method, then the cores were sequentially coated with the isolation layer Ⅰ, esomeprazole magnesium layer, isolation layer Ⅱ and enteric layer by bottom spray-type fluid-bed coating process to obtain the final product, the aspirin and esomeprazole magnesium co-loaded enteric-coated pellets. The results of in vitro test showed that the cumulative releases of two drugs from the pellets were below 5％ in 0.1 mol/L hydrochloric acid within two hours, while in pH 6.8 phosphate buffer, the cumulative releases of aspirin and esomeprazole magnesium were (5.9±1.1)％ and (78.5±1.4)％ for 15 min, (77.4±3.3)％ and (83.5±1.9)％ for 60 min, respectively. The pharmacokinetic test of the co-loaded enteric-coated pellets was carried out with rats as the models. The tmax, AUC0→∞, cmax and MRT0→∞ in rats after gavage administration were (1.50±0.00) and (3.50±0.50)h, (15.73±2.50) and (1 158.39±73.73)mg·L-1·h, (2.89±0.09) and (75.13±2.14)mg/L, (8.30±1.30) and (11.679±0.60)h respectively. There results showed that esomeprazole magnesium could burst release in pH 6.8 medium, while aspirin could release after a certain time lag, which was conducive to play a synergistic effect and reduced the gastrointestinal irritation of aspirin in long-term application.