Objective: To investigate and evaluate the anti-HBV activity of two kings of Matijin-su in vivo. Methods: Chronic HBV infection was established in C57BL/6J mice by injecting pAAV-HBV1.2 plasmid into high pressure caudal vein. Mice successfully modeled w.ere randomly divided into 4 groups, namely negative control group, positive control group, Matijin-su derivatives-1 group and Matijin-su derivatives-2 group, with 10 mice in each group. They were intraperitoneally injected with 200 μL volume of PBS, positive drug Birinapant 30 mg/kg, Matijin-su derivatives-1 (20 mg/kg) and Matijin-su derivatives-2 (20 mg/kg), respectively. Dosing was given once a week for 4 weeks. The anti-HBV effects of each group were observed before and after treatment, and the pathological morphology of liver tissue was detected after treatment. Results: After treatment, the serum levels of HBsAg, HBeAg, AST and ALT in group 1 and group 2 decreased significantly (P＜0.05). Compared with the positive control group, there was no significant difference in the degree of decline (P＞0.05). Histopathological comparison of liver tissue showed no significant difference among the groups. Conclusion: Matijin-su derivatives have some anti-HBV effects in vivo.