Search for new classes of HCV therapeutics by large scale screening of antiviral compounds and host cellular factors

2008 Fiscal Year Final Research Report

• Project/Area Number: 19390196
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: SAKAMOTO Naoya Tokyo Medical and Dental University, 大学院・医歯学総合研究科, 寄附講座准教授 (10334418)
• Co-Investigator(Kenkyū-buntansha): WATANABE Mamoru 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (10175127) ; CHEN Cheng-hsin 東京医科歯科大学, 医学部附属病院, 助教 (10376783) ; NAKAGAWA Mina 東京医科歯科大学, 大学院・医歯学総合研究科, 助教 (30401342) ; ITSUI Yasuhiro 東京医科歯科大学, 大学院・医歯学総合研究科, メディカルフェロー (20401341) ; KAKINUMA Sei 東京大学, 医科学研究所, 産学官連携研究員 (30372444) ; OSAJIMA Yuko 東京医科歯科大学, 医学部附属病院, 医員 (40451934) ; TASAKA Megumi 東京医科歯科大学, 医学部附属病院, 医員 (50510369)
• Co-Investigator(Renkei-kenkyūsha): WATANABE Mamoru 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (10175127) ; CHEN Cheng-hsin 東京医科歯科大学, 医学部附属病院, 助教 (10376783) ; NAKAGAWA Mina 東京医科歯科大学, 大学院・医歯学総合研究科, 助教 (30401342) ; ITSUI Yasuhiro 東京医科歯科大学, 大学院・医歯学総合研究科, メディカルフェロー (20401341) ; KAKINUMA Sei 東京大学, 医科学研究所, 産学官連携研究員 (30372444)
• Project Period (FY): 2007 – 2008
• Keywords: HCVレプリコン / HCV-JFH1培養系

Research Abstract

当該研究期間において我々は、HCV感染・増殖細胞モデルを用いて薬剤・生理活性化合物・短鎖ペプチドの抗ウイルス作用の網羅的スクリーニング、およびHCV増殖に関連する宿主因子の探索を遂行し、以下の結果を得た。(1) HCV-Feo replicon細胞を用いてDiversity-OrientedSynthesis(DOS)法で合成した8,000種の化合物のスクリーニングを施行し、HCV増殖を抑制する41種の化合物を同定し、構造活性相関解析にIC50の優れた5個のepoxide誘導体を同定した。(2) HCV複製増殖に関与する代謝・シグナルネットワークの網羅的解析により、脂質・コレステロール代謝に関わる代謝・シグナルネットワークの関与、関連薬剤の効果を確認した。(3) 漢方生薬成分化合物の細胞内HCV増殖に対する効果をHCVレプリコンシステムを用いて解析を行い、甘草由来の2種の化合物、isoliquiritigenin、及びglycycoumarinにHCV増殖抑制作用を見出した。今後これらの知見を統合し、HCV感染サイクルにおけるウイルスおよび宿主蛋白相互作用の細胞・分子レベルでの理解を深め、新規クラス抗HCV療法薬剤の開発・実用化に必要な基盤情報の蓄積を到達目標とし引き続き研究を遂行する。本研究で同定された化合物の作用機構の解析、小動物モデルを用いた効果・安全性の検証を進めることにより新たな抗ウイルス薬剤の開発につながると思われる。

Research Products

• [Journal Article] New therapeutic approaches to HCV.J (2009)
  • Author(s): Naoya Sakamoto, Mamoru Watanabe
  • Journal Title: Gastroenterol (in press)
  • Peer Reviewed
• [Journal Article] FTY720 suppresses the development of colitis in lymphoid-null mice by modulating the trafficking of colitogenic CD4+ T cellsin bone marrow (2009)
  • Author(s): Fujii T, Kanai T, Tomita T, Nemoto Y,Totsuka T, Sakamoto N, Nakamura T,Tsuchiya K, Okamoto R, Watanabe M
  • Journal Title: Eur J Immunol 38(12) Pages: 3290-3303
  • Peer Reviewed
• [Journal Article] Two flavonoids extracts from a herb, Glycyrrhizae radix, inhibit in-vitro hepatitis C virus replication (2009)
  • Author(s): Sekine-Osajima Y, Sakamoto N, Nakagawa M, Itsui Y, Tasaka M, Nishimura-Sakurai Y, Chen CH, Suda G, Mishima K, Onuki Y, Yamamoto M, Maekawa S, Enomoto N, Kanai T, Tsuchiya K, Watanabe M
  • Journal Title: Hepatology Res 39(1) Pages: 60-69
  • Peer Reviewed
• [Journal Article] Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA (2008)
  • Author(s): Sakamoto N, Tanabe Y, Yokota T, Saito K,Sekine-Osajima Y, Nakagawa M, Itsui Y,Tasaka M, Sakurai Y, Chen CH, Yano M,Ohkoshi S, Aoyagi Y, Maekawa S, Enomoto N, Kohara M, Watanabe M
  • Journal Title: J Gastro Hepatol 23(9) Pages: 1437-1447
  • Peer Reviewed
• [Journal Article] Effects of immunosuppressants on the progression ofhepatitis C in hepatitis C virus-positive renal transplantation and the usefulness of interferon therapy (2008)
  • Author(s): Nanmoku K, Imaizumi R, Tojimbara T,Nakajima I, Fuchinoue S, Sakamoto N, Watanabe M, Teraoka S
  • Journal Title: Transplant Proc 40(7) Pages: 2382-2385
  • Peer Reviewed
• [Journal Article] Griseofulvin, an oral antifungal agent, suppresses HCV replication in vitro (2008)
  • Author(s): Jin H, Yamashita A, Maekawa S, Yang P, HeL, Takayanagi S, Wakita T, Sakamoto N,Enomoto N, Ito M
  • Journal Title: Hepatology Res 38(9) Pages: 909-918
  • Peer Reviewed
• [Journal Article] Potential relevance of cytoplasmic viralsensors and related regulators involving innate immunity in antiviral response (2008)
  • Author(s): Asahina Y, Izumi N, Hirayama I, Tanaka T,Sato M, Yasui Y, Komatsu N, Umeda N,Hosokawa T, Ueda K, Tsuchiya K,Nakanishi H, Itakura J, Kurosaki M,Enomoto N, Tasaka M, Sakamoto N, Miyake S
  • Journal Title: Gastroenterology 134(5) Pages: 1396-1405
  • Peer Reviewed
• [Journal Article] Proteasomal degradation of Atoh1 by aberrant Wnt signaling maintains the undifferentiated state of colon cancer (2008)
  • Author(s): Aragaki M, Tsuchiya K, Okamoto R,Yoshioka S, Nakamura T, Sakamoto N,Kanai T, Watanabe M
  • Journal Title: Biochem Biophys Res Commun 368(4) Pages: 923-929
  • Peer Reviewed
• [Journal Article] Targeting lipid metabolism in the treatment of hepatitis C (2008)
  • Author(s): Amemiya F, Maekawa S, Itakura Y,Kanayama A, Takano S, Yamaguchi T,Itakura J, Kitamura T, Inoue T, Sakamoto M,Yamauchi K, Okada S, Sakamoto N, Enomoto N
  • Journal Title: J Infect Dis 197(3) Pages: 361-370
  • Peer Reviewed
• [Journal Article] Systemic, but not intestinal,IL-7 is essential for the persistence ofchronic colitis (2008)
  • Author(s): Tomita T, Kanai T, Nemoto Y, Totsuka T,Okamoto R, Tsuchiya K, Sakamoto N, Watanabe M
  • Journal Title: J immunol 180(1) Pages: 383-390
  • Peer Reviewed
• [Journal Article] Development of plaque assaysfor hepatitis C virus and isolation of mutantswith enhanced cytopathogenicity and replication capacity (2008)
  • Author(s): Sekine-Osajima Y, Sakamoto N(equal contribution), Nakagawa M, Itsui Y, TasakaM, Nishimura-Sakurai Y, Chen CH, Kanai T,Tsuchiya K, Wakita T, Enomoto N and Watanabe M
  • Journal Title: Virology 371 Pages: 71-85
  • Peer Reviewed
• [Journal Article] Identification of novel epoxide inhibitors of HCV replication、. a high-throughput screen (2007)
  • Author(s): Peng LF, Kim SS, Matchacheep S, Lei X, SuS, Lin W, Runguphan W, Choe WH, Sakamoto N, Ikeda M, Kato N, Beeler AB,Porco JA Jr, Schreiber SL and Chung RT
  • Journal Title: Antimicrob Agent Chemother 51(10) Pages: 3756-3759
  • Peer Reviewed
• [Journal Article] HCV nonstructural proteins responsible for suppression of RIG-I/Cardif-induced interferon response (2007)
  • Author(s): Tasaka M, Sakamoto N(equal contribution), Itakura Y, Nakagawa M, Itusi Y,Sekine-Osajima Y, Nishimura-Sakurai Y,Chen CH, Yoneyama M, Fujita T, Wakita T,Maekawa S, Enomoto N, Watanabe M
  • Journal Title: J Gen Virol 88 Pages: 3323-3333
  • Peer Reviewed
• [Journal Article] Bone morphogenetic protein-7 and interferon-alpha synergistically suppress hepatitis C virus replicon (2007)
  • Author(s): Sakamoto N, Yoshimura M, Kimura T, Toyama K, Sekine-Osajima Y, Watanabe M, Muramatsu M
  • Journal Title: Biochem Biophys Res Commun 357 Pages: 467-473
  • Peer Reviewed
• [Journal Article] A cell-based, high-throughput screen for small molecule regulators of HCV replication (2007)
  • Author(s): Kim SS, Peng LF, Lin W, Choe WH,Sakamoto N, Schreiber SL, Ikeda M, Kato N, Chung RT
  • Journal Title: Gastroenterology2007 132 Pages: 311-320
  • Peer Reviewed
• [Journal Article] Human Cord Blood Cells Transplanted into Chronic-Damaged Liver Exhibit Similar Characteristics to Functional Hepatocytes (2007)
  • Author(s): Kakinuma S, Asahina K, Okamura K,Teramoto K, Tateno C, Yoshizato K, TanakaY, Yasumizu T, Sakamoto N, Watanabe M,Teraoka H
  • Journal Title: Transplantation Proceedings 39 Pages: 240-243
  • Peer Reviewed
• [Journal Article] A functional genomic screen identifies cellular cofactors of hepatitis C virus replication
  • Author(s): Andrew W. Tai, Yair Benita, Lee F. Peng, Sun-Suk Kim, Naoya Sakamoto, Ramnik J.Xavier, Raymond T. Chung
  • Journal Title: Cell Host & Microbe (in press)
  • Peer Reviewed
• [Journal Article] Synergistic inhibition of intracellular hepatitis C virus replication by nelfinavir and interferon-α
  • Author(s): Satoshi Toma, Tsuyoshi Yamashiro, ShingoArakaki, Joji Shiroma, Tatsuji Maeshiro, Kenji Hibiya, Naoya Sakamoto, FukunoriKinjo, Masao Tateyama, Jiro Fujita
  • Journal Title: Journal of Viral Hepatitis (in press)
  • Peer Reviewed
• [Presentation] Establishment and genetic analyses of cytopatho genic HCV-JFH1 mutants by plaque-forming assay (2008)
  • Author(s): Kako Mishima, Naoya Sakamoto, YukoSekine-Osajima, Mina Nakagawa, MegumiTasaka, Yuki Nishimura-Sakurai, YasuhiroItsui, Takaji Wakita,and Mamoru Watanabe
  • Organizer: Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco,CA
  • Year and Date: 2008-11-01
• [Presentation] Suppression of interferon induction and response pathway by Hepatitis C virus NS4B (2008)
  • Author(s): M. Tasaka, N. Sakamoto, M. Nakagawa, Y.Itsui, Y. Nishimura-Sakurai, C. Chen, Goki Suda, Kako Mishima, Machi Yamamoto,Yuko Onuki, M. Watanabe
  • Organizer: Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco,CA
  • Year and Date: 2008-11-01
• [Presentation] Pretreatment assessment of PEG-interferon plus ribavirin anti-HCV therapy using viral genetic profiles, NS5A-ISDR and Core mutations (2008)
  • Author(s): Naoya Sakamoto, Mina Nakagawa,Cheng-Hsin Chen, Yasuhiro Itsui, Megumi Tasaka, Nobuyuki Enomoto,and Mamoru Watanabe
  • Organizer: Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco,CA
  • Year and Date: 2008-11-01
• [Presentation] THE ISOMERASE ACTIVITY OF CYCLOPHILIN A IS CRITICAL FOR HCV REPLICATION (2008)
  • Author(s): Bobardt M., Chatterji U., Selvarajah S., TangH., Sakamoto N., Coelmont L., Neyts J.,Dumont J.-M., Vuagniaux G.and Gallay P.
  • Organizer: 15th. International Meeting on Hepatitis C Virus & Related Viruses
  • Place of Presentation: San Antonio, TX
  • Year and Date: 2008-10-04
• [Presentation] Establishment and genetic analyses of cytopathogenic HCV-JFH1 mutants by plaque-forming assay (2008)
  • Author(s): Naoya Sakamoto, Kako Mishima, YukoSekine-Osajima, Mina Nakagawa, MegumiTasaka, Yuki Nishimura-Sakurai, YasuhiroItsui, Takaji Wakita,and Mamoru Watanabe
  • Organizer: 15th. International Meeting on Hepatitis C Virus & RelatedViruses
  • Place of Presentation: San Antonio, TX
  • Year and Date: 2008-10-04
• [Presentation] A novel NS5B-interacting protein modulates hepatitis C virus replication (2008)
  • Author(s): Jing-Tang Huang, Ching-Ping Tseng, Naoya Sakamoto, Shao-Chun Lu, Chuan-Mu Chen,Ju-Chien Cheng
  • Organizer: 15th. International Meeting on Hepatitis C Virus & Related Viruses
  • Place of Presentation: San Antonio, TX
  • Year and Date: 2008-10-04
• [Presentation] Antiviral effects of interferon-induced proteins, GBP-1, IFI6-16 and IFI27 and their interactions with hepatitis C virus nonstructural proteins (2007)
  • Author(s): Yasuhiro Itsui, Naoya Sakamoto, Mina Nakagawa, Yuko Sekine-Osajima, Megumi Tasaka,and Mamoru Watanabe
  • Organizer: Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: Boston, MA
  • Year and Date: 2007-10-31
• [Presentation] Plaque-forming assays for hepatitis C virus and isolation of HCV-JFH1 mutants with enhanced cytopathogenicity and replication capacity (2007)
  • Author(s): Naoya Sakamoto, Yuko Sekine-Osajima, Kako Mishima, Mina Nakagawa, Megumi Tasaka, Yuki Nishimura-Sakurai, YasuhiroItsui, Takaji Wakita,and Mamoru Watanabe
  • Organizer: Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: Boston, MA
  • Year and Date: 2007-10-31
• [Presentation] for the Ochanomizu Liver Conference : Clinical and virological factors that predict outcomes of interferon plus ribavirin combination therapy for HCV infection (2007)
  • Author(s): Sakamoto N, Nakagawa M, Watanabe M
  • Organizer: APDW 2007 Symposium
  • Place of Presentation: Kobe, Japan
  • Year and Date: 2007-10-18
• [Presentation] Development of plaque-forming assays for hepatitis C virus and isolation of mutants with enhanced cytopathogenicity and replication capacity (2007)
  • Author(s): Naoya Sakamoto, Yuko Sekine-Osajima, Kako Mishima, Mina Nakagawa, Megumi Tasaka, Yuki Nishimura-Sakurai, Yasuhiro Itsui, Takaji Wakita,and Mamoru Watanabe
  • Organizer: 14th. International Meeting on Hepatitis C Virus & Related Viruses
  • Place of Presentation: Glasgow, UK
  • Year and Date: 2007-09-09
• [Patent(Industrial Property Rights)] C型肝炎ウイルスのタンパク質合成及び/又はC型肝炎ウイルスの複製を抑制することができる二本鎖オリゴヌクレオチド (2003)
  • Inventor(s): 横田隆徳、榎本信幸、坂本直哉、多比良和誠、宮岸真
  • Industrial Property Rights Holder: 独立行政法人産業技術総合研究
  • Industrial Property Number: 特願2003-104940
  • Filing Date: 2003-05-11