Hepatitis C virus replication is inhibited by 22beta-methoxyolean-12-ene-3beta, 24(4beta)-diol (ME3738) through enhancing interferon-beta.
A derivative of soyasapogenol, 22beta-methoxyolean-12-ene-3beta, 24(4beta)-diol (ME3738), ameliorates liver injury induced by Concanavalin A in mice. We examined whether ME3738 has independent antiviral effects against hepatitis C virus (HCV) using an established HCV replication model that expresses the full-length genotype 1a HCV complementary DNA plasmid (pT7-flHCV-Rz) under the control of a replication-defective adenoviral vector expressing T7 polymerase. Hepatocellular carcinoma (HepG2) cells, human hepatoma (Huh7) cells, or monkey kidney (CV-1) cells were transfected with pT7-flHCV-Rz, and infected with adenoviral vector expressing T7 polymerase. ME3738 or interferon-alpha (IFN-alpha) was added thereafter and then protein and RNA were harvested from the cells at 9 days after infection. HCV-positive and HCV-negative strands were measured by real-time reverse-transcription polymerase chain reaction and HCV core protein expression was measured using an enzyme-linked immunosorbent assay. The messenger RNA levels of innate antiviral response-related genes were assessed using real-time reverse-transcription polymerase chain reaction. ME3738 dose-dependently reduced HCV-RNA and core protein in hepatocyte-derived cell lines. The antiviral effect was more pronounced in HepG2 than in Huh7 cells. ME3738 increased messenger RNA levels of interferon-beta (IFN-beta) and of IFN-stimulated genes (2'-5' oligoadenylate synthetase, myxovirus resistance protein A [MxA]). Interferon-beta knockdown by small interfering RNA abrogated the anti-HCV effect of ME3738. Moreover, the anti-HCV effects were synergistic when ME3738 was combined with IFN-alpha.


CONCLUSIONS
ME3738 has antiviral effects against HCV. The enhancement of autocrine IFN-beta suggests that ME3738 exerts antiviral action along the type I IFN pathway. This anti-HCV action by ME3738 was synergistically enhanced when combined with IFN-alpha. ME3738 might be a useful anti-HCV drug either with or without IFN-alpha.
Hiasa
Yoichi
Y
Department of Gastroenterology and Metabology, Ehime University Graduate School of Medicine, Ehime, Japan.
Kuzuhara
Hiroyuki
H
Tokumoto
Yoshio
Y
Konishi
Ichiro
I
Yamashita
Nobuyuki
N
Matsuura
Bunzo
B
Michitaka
Kojiro
K
Chung
Raymond T
RT
Onji
Morikazu
M
D000818:Animals; D000998:Antiviral Agents / Q000008:administration & dosage / Q000494:pharmacology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D018076:DNA, Complementary; D004259:DNA-Directed DNA Polymerase; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D019204:GTP-Binding Proteins / Q000235:genetics; D005838:Genotype; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D016899:Interferon-beta / Q000378:metabolism; D064547:Myxovirus Resistance Proteins; D009828:Oleanolic Acid / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D010957:Plasmids; D012367:RNA, Viral / Q000037:antagonists & inhibitors; D014162:Transfection; D015854:Up-Regulation; D014758:Viral Core Proteins / Q000037:antagonists & inhibitors; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D018076:DNA, Complementary; D016898:Interferon-alpha; C477463:ME3738; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D012367:RNA, Viral; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus; D009828:Oleanolic Acid; D016899:Interferon-beta; D004259:DNA-Directed DNA Polymerase; D019204:GTP-Binding Proteins