Objective To study the pharmacokinetics and bioequivalence of asparaginase (Asp) hyaluronic acid-graft-poly (ethylene glycol)/dimethyl-β-cyclodextrin (DCD) nanocapsules (AHDPs) in male Sprague-Dawley (SD) rats. Methods AHDPs were observed under the transmission electron microscope. The size, zeta potential and entrapment efficiency of AHDPs were examined. Asp activities were assayed after intravenous injection of AHDPs or free Asp in rats. Pharmacokinetic parameters were calculated by software DAS 2.1.1. Then the bioequivalence of AHDPs and free Asp were evaluated. Results The average particle size of AHDPs was (439.63±8.49) nm, zeta potential was (-20.43±2.20) mV, and entrapment efficiency was (55.75±4.11)% (n=3). AUC_{(0-48 h)} of AHDPs and free Asp were (138.93±0.89) U·mL^-1·h and (46.38±1.98) U·mL^-1·h, the AUC_0-∞ were (175.22±13.59) U·mL^-1·h and (51.44±3.01) U·mL^-1·h, and t_1/2 was (4.46±1.04) h and (1.86±0.38) h, respectively. Compared with free Asp, the AUC_{0-48 h}, AUC_0-∞ and t_1/2 of AHDPs were increased by 3.00, 3.40 and 2.40 times, respectively. The 90% confidential intervals of AUC_{0-48 h}, AUC_0-∞ and C_max were 76.9%-78.3%, 76.9%-78.3% and 92.8%-94.4%, respectively. Conclusion AHDPs can prolong the biological half-life and improve the bioavailability of Asp in rats. AHDPs and free Asp are not bioequivalent.