Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter December 14, 2019

Synergistic anti-hepatitis C virus activity of Ruta angustifolia extract with NS3 protein inhibitor

  • Tutik Sri Wahyuni EMAIL logo , Humairoh Mahfud , Adita Ayu Permatasari , Aty Widyawaruyanti and Achmad Fuad



Medicinal plants are known to perform many pharmacological actions due to their chemical metabolites, which include antiviral effects. Previously, the extract of Ruta angustifolia was shown to have potential anti-hepatitis C virus (HCV) activity without any cytotoxicity, with a 50% inhibitory concentration of 3.0 μg/mL and a 50% cytotoxicity concentration of >100 μg/mL. Furthermore, the combination of medicinal plants and current anti-HCV agents, such as a direct-acting antiviral agent, was shown to potentiate their overall effectiveness. In the course of this study, the ethanolic extract of R. angustifolia was evaluated for its anti-HCV effects; specifically, the mechanism of action on HCV NS3 and NS5A protease was investigated.


Analysis of the use of this extract in conjunction with current NS3 inhibitor drugs, simeprevir (SMV) and telaprevir (TVR), was performed. Anti-HCV activity was performed by in vitro culture of hepatocyte cells. The cells were infected and treated with various concentrations of the sample. HCV inhibition was calculated and CompuSyn software analysis was used to determine the synergistic effect of the combination.


Results demonstrated that R. angustifolia extract inhibited the post-entry step and decreased the protein levels of HCV NS3 and NS5A. The combination of extract and SMV and TVR mediated a synergistic effect.


These findings suggest that combining R. angustifolia extract with current anti-HCV drugs should be considered when developing alternative and complementary anti-HCV medicines.


The authors are sincerely grateful to Prof. Hak Hotta and Dr. Chie Aoki Utsubo for providing HCV and hepatocyte cells.

  1. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  2. Research funding: None declared.

  3. Competing interests: The authors state no conflict of interest.


[1] Wahyuni TS, Utsubo CA, Hotta H. Promising anti-hepatitis C virus compounds from natural resources. Nat Prod Commun 2016;11:1193–200.10.1177/1934578X1601100840Search in Google Scholar

[2] Pollio A, De Natale A, Appetiti E, Aliotta G, Touwaide A. Continuity and change in the Mediterranean medical tradition: Ruta spp. (rutaceae) in Hippocratic medicine and present practices. J Ethnopharmacol 2008;116:469–82.10.1016/j.jep.2007.12.013Search in Google Scholar PubMed

[3] Wahyuni TS, Widyawaruyanti A, Lusida MI, Fuad A, Soetjipto, Fuchino H, et al. Inhibition of hepatitis C virus replication by chalepin and pseudane IX isolated from Ruta angustifolia leaves. Fitoterapia 2014;99:276–83.10.1016/j.fitote.2014.10.011Search in Google Scholar PubMed

[4] Pawlotsky JM, Negro F, Aghemo A, Berenguer M, Dalgard O, Dusheiko G, et al. EASL recommendations on treatment of hepatitis C 2018. J Hepatol 2018;69:461–511.10.1016/j.jhep.2018.03.026Search in Google Scholar PubMed

[5] Manns MP, Foster GR, Rockstroh JK, Zeuzem S, Zoulim F, Houghton M. The way forward in HCV treatment – finding the right path. Nat Rev Drug Discov 2007;6:991–1000.10.1038/nrd2411Search in Google Scholar PubMed

[6] Gonzalez-Grande R, Jimenez-Perez M, Gonzalez Arjona C, Mostazo Torres J. New approaches in the treatment of hepatitis C. World J Gastroenterol 2016;22:1421–32.10.3748/wjg.v22.i4.1421Search in Google Scholar PubMed PubMed Central

[7] Ashfaq UA, Masoud MS, Nawaz Z, Riazuddin S. Glycyrrhizin as antiviral agent against hepatitis C virus. J Transl Med 2011;9:112.10.1186/1479-5876-9-112Search in Google Scholar PubMed PubMed Central

[8] Lee JC, Chen WC, Wu SF, Tseng CK, Chiou CY, Chang FR, et al. Anti-hepatitis C virus activity of Acacia confusa extract via suppressing cyclooxygenase-2. Antiviral Res 2011;89:35–42.10.1016/j.antiviral.2010.11.003Search in Google Scholar PubMed

[9] Wahyuni TS, Tumewu L, Permanasari AA, Apriani E, Adianti M, Rahman A, et al. Antiviral activities of Indonesian medicinal plants in the East Java region against hepatitis C virus. Virol J 2013;10:259.10.1186/1743-422X-10-259Search in Google Scholar PubMed PubMed Central

[10] Chou TC. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 2006;58:621–81.10.1124/pr.58.3.10Search in Google Scholar PubMed

[11] Tallarida RJ. An overview of drug combination analysis with isobolograms. J Pharmacol Exp Ther 2006;319:1–7.10.1124/jpet.106.104117Search in Google Scholar PubMed

[12] Tallarida RJ. Drug combinations: tests and analysis with isoboles. Curr Prot Pharmacol 2016;72:9.19.1–19.10.1002/0471141755.ph0919s72Search in Google Scholar PubMed PubMed Central

[13] Ciesek S, von Hahn T, Colpitts CC, Schang LM, Friesland M, Steinmann J, et al. The green tea polyphenol, epigallocatechin-3-gallate, inhibits hepatitis C virus entry. Hepatology 2011;54:1947–55.10.1002/hep.24610Search in Google Scholar PubMed

[14] Khachatoorian R, Arumugaswami V, Raychaudhuri S, Yeh GK, Maloney EM, Wang J, et al. Divergent antiviral effects of bioflavonoids on the hepatitis C virus life cycle. Virology 2012;433:346–55.10.1016/j.virol.2012.08.029Search in Google Scholar PubMed PubMed Central

[15] Dai J, Carte BK, Sidebottom PJ, Sek Yew AL, Ng S, Huang Y, et al. Circumdatin G, a new alkaloid from the fungus Aspergillus ochraceus. J Nat Prod 2001;64:125–6.10.1021/np000381uSearch in Google Scholar PubMed

[16] Elsebai MF, Koutsoudakis G, Saludes V, Perez-Vilaro G, Turpeinen A, Mattila S, et al. Pan-genotypic hepatitis C virus inhibition by natural products derived from the wild Egyptian artichoke. J Virol 2016;90:1918–30.10.1128/JVI.02030-15Search in Google Scholar PubMed PubMed Central

[17] Tamori A, Enomoto M, Kawada N. Recent advances in antiviral therapy for chronic hepatitis C. Mediat Inflamm 2016;2016:11.10.1155/2016/6841628Search in Google Scholar PubMed PubMed Central

[18] Izquierdo L, Helle F, Francois C, Castelain S, Duverlie G, Brochot E. Simeprevir for the treatment of hepatitis C virus infection. Pharmgenomics Pers Med 2014;7:241–9.10.2147/PGPM.S52715Search in Google Scholar

[19] Sarrazin C, Zeuzem S. Resistance to direct antiviral agents in patients with hepatitis C virus infection. Gastroenterology 2010;138:447–62.10.1053/j.gastro.2009.11.055Search in Google Scholar PubMed

Received: 2019-11-17
Accepted: 2019-11-18
Published Online: 2019-12-14

© 2019 Walter de Gruyter GmbH, Berlin/Boston

Downloaded on 2.6.2023 from
Scroll to top button