The target compounds were obtained through amino protection, acylation, condensation, deprotection and salification of tenofovir as the starting material, eight compounds were synthesized(Ⅲa～Ⅲh). The effect of different alkaline reagents on the conversion of compound Ⅰ during the preparation of compound Ⅱ and ratio of acetic acid to prodrug parent was studied. The optimal conditions were as following: Triethylamine was selected as the alkaline reagent; The molar ratio of acetic acid to prodrug parent was 0.74～0.93∶1. The structures of target products were confirmed by ESI-MS and 1HNMR. The synthesis method was simple and the yield of eight compounds was from 16.1% to 40.7%. A potential compound Ⅲh (tenofovir dihydroxyacetone ethylene ketal) was screened by preliminary activity study. Compared with tenofovir dipivoxil fumarate(TDF), the stability of compound Ⅲh in rat and human plasma in vitro were respectively increased by 2-fold and 2.5-fold. The live and kidney PMPA AUC(0-t) of compound Ⅲh in mice in vivo were respectively (47314.75±10128.11) (μg•h)/L and (21670.64±8964.98) (μg•h)/L while the AUC(0-t) of TDF were respectively (213269.79±10750.47) (μg•h)/L and (46379.24±3944.65) (μg•h)/L. This prodrug showed a certain liver targeting and low nephrotoxicity, which was worthy of further research.