Patil Vishal S, Harish Darasaguppe R, Charla Rajitha, Bhandare Vishwambhar Vishnu, Gujarathi Swarup S, Beerwala Faizan A, Patil Priyanka P, Jalalpure Sunil S, Hegde Harsha V, Roy Subarna
ICMR-National Institute of Traditional Medicine, Nehru Nagar, Belagavi, Karnataka, 590010, India.
KLE College of Pharmacy, Belagavi, KLE Academy of Higher Education and Research, Nehru Nagar, Belagavi,, Karnataka, 590010, India.
BMC Complement Med Ther. 2025 Mar 8;25(1):95. doi: 10.1186/s12906-025-04807-z.
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.
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.
AP exhibits lower cytotoxicity (CC = 832.915 µg/mL) than TP (CC = 593.122 µg/mL) after 24 h, with Tenofovir disoproxil fumarate (TDF) showing minimal cytotoxicity (CC > 500 µM). Both AP and TP significantly decreased intracellular HBV DNA with a > 2 fold reduction at higher concentrations (125-500 µg/mL) but had no significant effect on pgRNA level. AP and TP 500 µg/mL effectively inhibited HBsAg secretion (95% and 80% inhibition, respectively), over 120 h. AP also showed inhibition of HBeAg secretion (75-82%), while TP exhibited a higher inhibition of 90% at 24 h. TDF showed consistent but lower inhibitory effects on HBsAg and HBeAg. The HBx-HBXIP interaction inhibition assay showed AP's greater inhibitory capacity (IC < 62.5 µg/mL) compared to TP (IC = 806.69 µg/mL). Computational studies further validated these findings, showing stable binding interactions of AP compounds (flavonoids) with HBx protein (with Arg138 and His139, Lys140, and Trp141 residues participating in the interaction with HBXIP), corroborating their potential in disrupting HBV replication. Molecular dynamics simulations confirmed the stability of these interactions over 100ns.
AP exhibits potent anti-HBV activities, making it a promising candidate for further therapeutic development.
乙型肝炎病毒（HBV）的X蛋白（HBx）在HBV发病机制中起关键作用，然而，目前诸如HIV逆转录酶（RT）抑制剂这类因活性位点相似而靶向HBV RT的治疗方法，具有严重的副作用、耐药风险和高昂成本。本研究通过计算分析，研究穿心莲（AP）和杨叶肖槿（TP）对表达HBV的HepG2.2.15细胞的抗乙型肝炎病毒（HBV）特性。
进行体外细胞毒性、逆转录酶抑制、通过qRT-PCR进行DNA和pgRNA定量、HBsAg和HBeAg的时间进程分析以及HBX-HBXIP相互作用抑制研究。通过分子对接和动力学研究分析HBX与HBXIP的相互作用，以及植物化合物与HBx的相互作用。
24小时后，AP的细胞毒性（CC = 832.915μg/mL）低于TP（CC = 593.122μg/mL），替诺福韦酯（TDF）的细胞毒性最小（CC＞500μM）。AP和TP在较高浓度（125 - 500μg/mL）时均显著降低细胞内HBV DNA，降低幅度超过2倍，但对pgRNA水平无显著影响。AP和TP 500μg/mL在120小时内有效抑制HBsAg分泌（分别为95%和80%抑制）。AP还显示出对HBeAg分泌的抑制作用（75 - 82%），而TP在24小时时的抑制率更高，为90%。TDF对HBsAg和HBeAg显示出持续但较低的抑制作用。HBx-HBXIP相互作用抑制试验表明，与TP（IC = 806.69μg/mL）相比，AP具有更强的抑制能力（IC＜62.5μg/mL）。计算研究进一步验证了这些发现，表明AP化合物（黄酮类）与HBx蛋白具有稳定的结合相互作用（Arg138、His139、Lys140和Trp141残基参与与HBXIP的相互作用），证实了它们在破坏HBV复制方面的潜力。分子动力学模拟证实了这些相互作用在100ns内具有稳定性。
AP具有强大的抗HBV活性，使其成为进一步治疗开发的有希望的候选药物。
