BACKGROUND: The HBx protein of hepatitis B virus (HBV) plays a crucial role in HBV pathogenesis, yet current treatments like HIV reverse transcriptase (RT) inhibitors, which target HBV RT due to similar active sites, have severe side effects, risk of drug resistance, and high costs. The present study investigates the anti-hepatitis B virus (HBV) properties of Andrographis paniculata (AP) and Thespesia populnea (TP) on HBV expressing HepG2.2.15 cells and by computational analysis. METHODS: In vitro cytotoxicity, reverse transcriptase inhibitory, DNA and pgRNA quantification by qRT-PCR, time course analysis of HBsAg and HBeAg, and HBX-HBXIP interaction inhibition studies were conducted. The interaction of HBX with HBXIP, and phytocompounds' interaction with HBx was analyzed through molecular docking and dynamics studies. RESULTS: AP exhibits lower cytotoxicity (CC CONCLUSIONS: AP exhibits potent anti-HBV activities, making it a promising candidate for further therapeutic development.