title: Identification of Antiviral Agents Targeting Hepatitis B Virus Promoter from Extracts of Indonesian Marine Organisms by a Novel Cell-Based Screening Assay.
abstract: The current treatments of chronic hepatitis B (CHB) face a limited choice of vaccine, antibody and antiviral agents. The development of additional antiviral agents is still needed for improvement of CHB therapy. In this study, we established a screening system in order to identify compounds inhibiting the core promoter activity of hepatitis B virus (HBV). We prepared 80 extracts of marine organisms from the coral reefs of Indonesia and screened them by using this system. Eventually, two extracts showed high inhibitory activity (>95%) and low cytotoxicity (66% to 77%). Solvent fractionation, column chromatography and NMR analysis revealed that 3,5-dibromo-2-(2,4-dibromophenoxy)-phenol (compound 1) and 3,4,5-tribromo-2-(2,4-dibromophenoxy)-phenol (compound 2), which are classified as polybrominated diphenyl ethers (PBDEs), were identified as anti-HBV agents in the extracts. Compounds 1 and 2 inhibited HBV core promoter activity as well as HBV production from HepG2.2.15.7 cells in a dose-dependent manner. The EC50 values of compounds 1 and 2 were 0.23 and 0.80 µM, respectively, while selectivity indexes of compound 1 and 2 were 18.2 and 12.8, respectively. These results suggest that our cell-based HBV core promoter assay system is useful to determine anti-HBV compounds, and that two PBDE compounds are expected to be candidates of lead compounds for the development of anti-HBV drugs.
authors.affiliation: Department of Microbiology, Division of Medical Sciences, Graduate School of Interdisciplinary Research, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. atsuyay@yamanashi.ac.jp.
authors.forename: Atsuya
authors.initials: A
authors.lastname: Yamashita
authors.affiliation: Department of Microbiology, Division of Medical Sciences, Graduate School of Interdisciplinary Research, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. yfujimoto@greiner-bio-one.co.jp.
authors.forename: Yuusuke
authors.initials: Y
authors.lastname: Fujimoto
authors.affiliation: Department of Chemistry, Biology and Marine Science, University of the Ryukyus, 1 Senbaru, Nishihara, Okinawa 903-0213, Japan. m.tamaki.4@jaist.ac.jp.
authors.forename: Mayumi
authors.initials: M
authors.lastname: Tamaki
authors.affiliation: Department of Chemistry, Faculty of Science, Lampung University, Jl. Sumantri Brodjonegoro No. 1, Bandar Lampung 35145, Indonesia. andi.setiawan@fmipa.unila.ac.id.
authors.forename: Andi
authors.initials: A
authors.lastname: Setiawan
authors.affiliation: Department of Microbiology, Division of Medical Sciences, Graduate School of Interdisciplinary Research, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. tomohisat@yamanashi.ac.jp.
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Department of Microbiology, Division of Medical Sciences, Graduate School of Interdisciplinary Research, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. kaorid@yamanashi.ac.jp.
authors.forename: Kaori
authors.initials: K
authors.lastname: Okuyama-Dobashi
authors.affiliation: Department of Microbiology, Division of Medical Sciences, Graduate School of Interdisciplinary Research, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. hirotake@yamanashi.ac.jp.
authors.forename: Hirotake
authors.initials: H
authors.lastname: Kasai
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. kwatashi@nih.go.jp.
authors.forename: Koichi
authors.initials: K
authors.lastname: Watashi
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. wakita@nih.go.jp.
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
authors.affiliation: Division of Antiviral Chemotherapy Center for Chronic Viral Disease, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8544, Japan. toyama@m2.kufm.kagoshima-u.ac.jp.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Toyama
authors.affiliation: Division of Antiviral Chemotherapy Center for Chronic Viral Disease, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8544, Japan. m-baba@m2.kufm.kagoshima-u.ac.jp.
authors.forename: Masanori
authors.initials: M
authors.lastname: Baba
authors.affiliation: Naturalis, National Museum of Natural History, P.O. Box 9517, Leiden 2300 RA, The Netherlands. Nicole.devoogd@naturalis.nl.
authors.forename: Nicole J
authors.initials: NJ
authors.lastname: de Voogd
authors.affiliation: The First Department of Internal Medicine, Faculty of Medicine, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. maekawa@yamanashi.ac.jp.
authors.forename: Shinya
authors.initials: S
authors.lastname: Maekawa
authors.affiliation: The First Department of Internal Medicine, Faculty of Medicine, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. enomoto@yamanashi.ac.jp.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Enomoto
authors.affiliation: Department of Chemistry, Biology and Marine Science, University of the Ryukyus, 1 Senbaru, Nishihara, Okinawa 903-0213, Japan. jtanaka@sci.u-ryukyu.ac.jp.
authors.forename: Junichi
authors.initials: J
authors.lastname: Tanaka
authors.affiliation: Department of Microbiology, Division of Medical Sciences, Graduate School of Interdisciplinary Research, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. kmoriishi@yamanashi.ac.jp.
authors.forename: Kohji
authors.initials: K
authors.lastname: Moriishi
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D059001:Aquatic Organisms / Q000378:metabolism*; D045744:Cell Line, Tumor; D057894:Coral Reefs; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics; D057166:High-Throughput Screening Assays; D006801:Humans; D007214:Indonesia; D011401:Promoter Regions, Genetic
chemical_list: D000998:Antiviral Agents
keywords: HBV; HBV core promoter; antiviral agent; hepatitis B virus; high-throughput screening; marine organism