Background: Over 350 million people worldwide are infected with hepatitis B virus (HBV),a major cause of liver failure and hepatocellular carcinoma.Current therapeutic agents are highly effective,but are also associated with development of viral resistance.Therefore,strategies for identifying other anti-HBV agents with specific,but distinctive mechanisms of action are needed.The human La (hLa) protein,which forms a stabilizing complex with HBV RNA ribonucleoprotein to promote HBV replication,is a promising target of molecular therapy.Aims: This study aimed to discover novel inhibitors of hLa that could inhibit HBV replication and expression.Methods: A multistage molecular docking approach was used to screen a Specs database and an in-house library against hLa binding sites.Sequential in vitro evaluations were performed to detect potential compounds with high scores in HepG2.2.15 cells.Results: Of the 26 potential compounds with high scores chosen for experimental verification,12 had HBV DNA inhibition ratios of less than 50% with P<0.05.Six had significant inhibition of HBV e antigen (HBeAg) levels,and 13 had significant inhibition of HBV surface antigen (HBsAg) levels by in vitro assays.Compounds HBSC-11,HBSC-15 and HBSC-34 (HBSC is system prefix for active compounds screened by the library) were selected for evaluation.HBSC-11 was found to have an obvious inhibitory effect on hLa transcription and expression.Conclusions: Our findings suggest that anti-HBV activity of HBSC-11 may be mediated by a reduction in hLa levels.In addition,our data suggest the potential clinical use of hLa inhibitors,such as HBSC-11,for treating HBV infection.