Ginsenosides are a class of natural steroid glycosides and triterpene saponins found in Panax ginseng. After screening of a commercial ginsenoside compound library for low cellular cytotoxicity and the ability to mediate efficient reductions in hepatitis B virus (HBV) mRNA expression levels in HepG2.2.15 cells, three ginsenosides (Rg6, Rh4, and Rb3) are selected. Thereafter, using the same cellular model, all three ginsenosides are shown to mediate efficient, selective inhibition of HBV mRNA expression levels, and also interfere with the secretion of both HBV particles and hepatitis B surface antigen (HBsAg). Drug combination studies are performed in both HepG2.2.15 and HBV-infected HepG2-NTCPsec+ cell models with the selected ginsenosides and lamivudine (LMV), a nucleoside analogue used to treat chronic hepatitis B (CHB) infections. These studies, involving RT-qPCR and ELISA, suggest that Rh4/LMV combinations in particular act synergistically to inhibit the secretion of HBV particles and HBsAg. Therefore, on the assumption that appropriate in vivo data are in future agreement, Rh4, in particular, might be used in combination with nucleoside/nucleotide analogues (NUCs) to devise an effective, cost-efficient combination therapy for the treatment of patients with CHB infections.

Applied Molecular Virology Laboratory Institut Pasteur Korea 696 Sampyeong dong Bundang gu Seongnam si Gyeonggi do South Korea

Department of Chemistry and Biochemistry Mendel University in Brno Zemědělská 1665 1 CZ 613 00 Brno Czech Republic

Department of Experimental Biology Faculty of Science Masaryk University Kamenice 735 5 CZ 625 00 Brno Czech Republic

Division of Bio Medical Science and Technology University of Science and Technology Daejeon South Korea

Institute of Parasitology Biology Centre of the Czech Academy of Sciences Branišovská 1160 31 CZ 370 05 České Budějovice Czech Republic

KP Therapeutics s r o Purkyňova 649 127 CZ 612 00 Brno Czech Republic

Screening Discovery Platform Institut Pasteur Korea 696 Sampyeong dong Bundang gu Seongnam si Gyeonggi do South Korea

Veterinary Research Institute Hudcova 296 70 CZ 621 00 Brno Czech Republic

See more in PubMed

Arbab A.H., Parvez M.K., Al-Dosari M.S., Al-Rehaily A.J. In vitro evaluation of novel antiviral activities of 60 medicinal plants extract against hepatitis B virus. Exp. Ther. Med. 2017;14:626–634. PubMed PMC

Attele A.S., Wu J.A., Yuan C.S. Ginseng pharmacology: multiple constituents and multiple actions. Biochem. Pharmacol. 1999;58:1685–1693. PubMed

Bouredja N., Bouthiba M., Kebir M. Ethnobotanical study of medicinal plants used by herbalists for the treatment of respiratory diseases in the region of Oran, Algeria. Br. J. Med. Health Sci. 2020;2:92–97.

Caly L., Druce J.D., Catton M.G., Jans D.A., Wagstaff K.M. The FDA-approved drug ivermectin inhibits the replication of SARS-CoV-2 in vitro. Antivir. Res. 2020;178 PubMed PMC

Chen Y., Zhu J. Anti-HBV effect of individual traditional Chinese herbal medicine in vitro and in vivo: an analytic review. J. Viral Hepat. 2013;20:445–452. PubMed

Cheng Z., Sun G., Guo W., Huang Y., Sun W., Zhao F., Hu K. Inhibition of hepatitis B virus replication by quercetin in human hepatoma cell lines. Virol. Sin. 2015;30:261–268. PubMed PMC

Chou T.C. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol. Rev. 2006;58:621–681. PubMed

Chou T.C. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010;70:440–446. PubMed

Chou S.C., Huang T.J., Lin E.H., Huang C.H., Chou C.H. Anti-hepatitis B virus constituents of Solanum erianthum. Nat. Prod. Commun. 2012;7:153–156. PubMed

Duraisamy G.S., Bhosale D., Lipenská I., Huvarova I., Růžek D., Windisch M.P., Miller A.D. Advanced therapeutics, vaccinations, and precision medicine in the treatment and management of chronic hepatitis B viral infections; where are we and where are we going? Viruses. 2020;12:998. PubMed PMC

Farrell G.C., Teoh N.C. Management of chronic hepatitis B virus infection: a new era of disease control. Intern. Med. J. 2006;36:100–113. PubMed

Glatthaar-Saalmüller B., Rauchhaus U., Rode S., Haunschild J., Saalmüller A. Antiviral activity in vitro of two preparations of the herbal medicinal product Sinupret® against viruses causing respiratory infections. Phytomedicine. 2011;19:1–7. PubMed PMC

Goh V.S.L., Mok C.K., Chu J.J.H. Antiviral natural products for arbovirus infections. Molecules. 2020;25:2796. PubMed PMC

Helfer M., Koppensteiner H., Schneider M., Rebensburg S., Forcisi S., Müller C., Schmitt-Kopplin P., Schindler M., Brack-Werner R. The root extract of the medicinal plant Pelargonium sidoides is a potent HIV-1 attachment inhibitor. PLoS One. 2014;9 PubMed PMC

Hong C., Yang P., Li S., Guo Y., Wang D., Wang J. In vitro/in vivo metabolism of ginsenoside Rg5 in rat using ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry. Molecules. 2018;23:2113. PubMed PMC

Huang Q., Zhang S., Huang R., Wei L., Chen Y., Lv S., Liang C., et al. Isolation and identification of an anti-hepatitis B virus compound from Hydrocotyle sibthorpioides Lam. J. Ethnopharmacol. 2013;150:568–575. PubMed

Huang J., Su D., Feng Y., Liu K., Song Y. Antiviral herbs--present and future. Infect. Disord.: Drug Targets. 2014;14:61–73. PubMed

Huang S., Liu Y., Zhang Y., Zhang R., Zhu C., Fan L., Pei G., Zhang B., Shi Y. Baicalein inhibits SARS-CoV-2/VSV replication with interfering mitochondrial oxidative phosphorylation in a mPTP dependent manner. Signal Transduct. Targeted Ther. 2020;5:266. PubMed PMC

Kang L.J., Choi Y.J., Lee S.G. Stimulation of TRAF6/TAK1 degradation and inhibition of JNK/AP-1 signalling by ginsenoside Rg3 attenuates hepatitis B virus replication. Int. J. Biochem. Cell Biol. 2013;45:2612–2621. PubMed

Kim S.J., Jang J.Y., Kim E.J., Cho E.K., Ahn D.G., Kim C., Park H.S., et al. Ginsenoside Rg3 restores hepatitis C virus-induced aberrant mitochondrial dynamics and inhibits virus propagation. Hepatology. 2017;66:758–771. PubMed PMC

Kim S.S., Ahn E.K., Cho S.Y., Park R.W., Cho H.J., Kim J.H., Kim H.G., et al. Impact of nucleos(t)ide analog treatment on the development of malignancy in patients with chronic hepatitis B. Medicine (Baltim.) 2018;97 PubMed PMC

König A., Yang J., Jo E., Park K.H.P., Kim H., Than T.T., Song X., Qi X., Dai X., et al. Efficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells. J. Hepatol. 2019;71:289–300. PubMed

Lee S., Lee M.S., Kim C.T., Kim I.H., Kim Y. Ginsenoside Rg3 reduces lipid accumulation with AMP-activated protein kinase (AMPK) activation in HepG2 cells. Int. J. Mol. Sci. 2012;13:5729–5739. PubMed PMC

Lee M.H., Lee B.H., Lee S., Choi C. Reduction of hepatitis A virus on FRhK-4 cells treated with Korean red ginseng extract and ginsenosides. J. Food Sci. 2013;78:M1412–M1415. PubMed

León P., Pozo F., Echevarría J.M. Detection of hepatitis B virus variants resistant to lamivudine and famciclovir among randomly selected chronic carriers from Spain. Enferm. Infecc. Microbiol. Clín. 2004;22:133–137. PubMed

Li J., Meng A.P., Guan X.L., Li J., Wu Q., Deng S.P., Su X.J., et al. Anti-hepatitis B virus lignans from the root of Streblus asper. Bioorg. Med. Chem. Lett. 2013;23:2238–2244. PubMed

Liang Y.Y., Wang B., Qian D.M., Li L., Wang Z.H., Hu M., Song X.X. Inhibitory effects of Ginsenoside Rb1 on apoptosis caused by HSV-1 in human glioma cells. Virol. Sin. 2012;27:19–25. PubMed PMC

Livak K.J., Schmittgen T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25:402–408. PubMed

Lok A.S., Zoulim F., Dusheiko G., Ghany M.G. Hepatitis B cure: from discovery to regulatory approval. Hepatology. 2017;66:1296–1313. PubMed PMC

Lü J.M., Yao Q., Chen C. Ginseng compounds: an update on their molecular mechanisms and medical applications. Curr. Vasc. Pharmacol. 2009;7:293–302. PubMed PMC

Musarra-Pizzo M., Pennisi R., Ben-Amor I., Mandalari G., Sciortino M.T. Antiviral activity exerted by natural products against human viruses. Viruses. 2021;13:828. PubMed PMC

Parvez M.K., Arab A.H., Al-Dosari M.S., Al-Rehaily A.J. Antiviral natural products against chronic hepatitis B: recent developments. Curr. Pharmaceut. Des. 2016;22:286–293. PubMed

Qiu L.P., Chen K.P. Anti-HBV agents derived from botanical origin. Fitoterapia. 2013;84:140–157. PubMed

Ryu Y.B., Jeong H.J., Kim J.H., Kim Y.M., Park J.Y., Kim D., Nguyen T.T., et al. Biflavonoids from Torreya nucifera displaying SARS-CoV 3CL(pro) inhibition. Bioorg. Med. Chem. 2010;18:7940–7947. PubMed PMC

Sagar S., Kaur M., Minneman K.P. Antiviral lead compounds from marine sponges. Mar. Drugs. 2010;8:2619–2638. PubMed PMC

Sanna G., Farci P., Busonera B., Murgia G., La Colla P., Giliberti G. Antiviral properties from plants of the Mediterranean flora. Nat. Prod. Res. 2015;29:2065–2070. PubMed

Serkedjieva J., Nikolova E., Kirilov N. Synergistic inhibition of influenza A virus replication by a plant polyphenol-rich extract and epsilon-aminocaproic acid in vitro and in vivo. Acta Virol. 2010;54:137–145. PubMed

Shaikh T., Cooper C. Reassessing the role for lamivudine in chronic hepatitis B infection: a four-year cohort analysis. Can. J. Gastroenterol. 2012;26:148–150. PubMed PMC

Simões C.M., Falkenberg M., Schenkel Mentz L.A., Amoros E.P., Girre M. Antiviral activity of south Brazilian medicinal plant extracts. Phytomedicine. 1999;6:205–214. PubMed

Song M.Y., Kim B.S., Kim H. Influence of Panax ginseng on obesity and gut microbiota in obese middle-aged Korean women. J. Ginseng Res. 2014;38:106–115. PubMed PMC

Sells M.A., Chen M.L., Acs G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc. Natl. Acad. Sci. U.S.A. 1987;84:1005–1009. PubMed PMC

Tavakolpour S., Darvishi M., Mirsafaei H.S., Ghasemiadl M. Nucleoside/nucleotide analogues in the treatment of chronic hepatitis B infection during pregnancy: a systematic review. Inf. Disp. 2018;50:95–106. PubMed

Tian Y., Sun L.M., Liu X.Q., Li B., Wang Q., Dong J.X. Anti-HBV active flavone glucosides from Euphorbia humifusa Willd. Fitoterapia. 2010;81:799–802. PubMed

Watanabe Y., Yamamoto H., Oikawa R., Toyota M., Yamamoto M., Kokudo N., Tanaka S., et al. DNA methylation at hepatitis B viral integrants is associated with methylation at flanking human genomic sequences. Genome Res. 2015;25:328–337. PubMed PMC

Wu Y.H. Naturally derived anti-hepatitis B virus agents and their mechanism of action. World J. Gastroenterol. 2016;22:188–204. PubMed PMC

Xiao Y., Liu C., Tang W., Zhang H., Chen X. Evans blue inhibits HBV replication through a dual antiviral mechanism by targeting virus binding and capsid assembly. Front. Microbiol. 2019;10:2638. PubMed PMC

Xu R., Hu P., Li Y., et al. Advances in HBV infection and replication systems in vitro. Virol. J. 2021;18:105. PubMed PMC

Yan H., Zhong G., Xu G., He W., Jing Z., Gao Z., Huang Y., et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. Elife. 2012;1 PubMed PMC

Yu J.S., Roh H.S., Baek K.H., Lee S., Kim S., So H.M., Moon E., et al. Bioactivity-guided isolation of ginsenosides from Korean Red Ginseng with cytotoxic activity against human lung adenocarcinoma cells. J. Ginseng Res. 2018;42:562–570. PubMed PMC

Zandi K., Musall K., Oo A., Cao D., Liang B., Hassandarvish P., Lan S., Slack R.L., Kirby K.A., Bassit L., Amblard F., Kim B., AbuBakar S., Sarafianos S.G., Schinazi R.F. Baicalein and baicalin inhibit SARS-CoV-2 RNA-dependent-RNA polymerase. Microorganisms. 2021;9:893. PubMed PMC

Zhang J., Li W., Yuan Q., Zhou J., Zhang J., Cao Y., Fu G., Hu W. Transcriptome analyses of the anti-proliferative effects of 20(S)-ginsenoside Rh2 on HepG2 cells. Front. Pharmacol. 2019;10:1331. PubMed PMC

Zhang Y.B., Zhang X.L., Chen N.H., Wu Z.N., Ye W.C., Li Y.L., Wang G.C. Four matrine-based alkaloids with antiviral activities against HBV from the seeds of Sophora alopecuroides. Org. Lett. 2017;19:424–427. PubMed

Zhao L., Ma Y., Chen C., Liu S., Wu W. Pharmacokinetic and metabolic studies of ginsenoside Rb3 in rats using RRLC-Q-TOF-MS. J. Chromatogr. Sci. 2018;56:480–487. PubMed

Zhou X., Liu J., Yang B., Lin X., Yang X.W., Liu Y. Marine natural products with anti-HIV activities in the last decade. Curr. Med. Chem. 2013;20:953–973. PubMed

Zhou N.J., Geng C.A., Huang X.Y., Ma Y.B., Zhang X.M., Wang J.L., Chen J.J. Anti-hepatitis B virus active constituents from Swertia chirayita. Fitoterapia. 2015;100:27–34. PubMed