宿主・ウイルスを標的とした抗HCV薬剤・化合物の大規模スクリーニング

2011 Fiscal Year Annual Research Report

• Project/Area Number: 21390226
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 坂本 直哉 東京医科歯科大学, 大学院・医歯学総合研究科, 寄附講座教員 (10334418)
• Co-Investigator(Kenkyū-buntansha): 中川 美奈 東京医科歯科大学, 医学部附属病院, 助教 (30401342) ; 東 正新 東京医科歯科大学, 医学部附属病院, 助教 (10376783) ; 渡辺 守 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (10175127)
• Keywords: C型肝炎 / レプリコンシステム

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-Oriented Synthesis(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個のエポキシ誘導体を同定した。さらに新たな化学合成ライブラリーの4,000種の化合物をスクリーニングし、HCV replicon増殖、HCV-JFH1感染を抑制する5種の化合物を同定し、構造類似化合物のスクリーニングを追加し構造活性相関があることを確認した。(3)SRタンパク質特異的キナーゼを標的とした小分子ライブラリーのスクリーニングを行い、特異的な抗HCV活性を持つ分子4個を同定した。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] Identification of novel N-(morpholine-4-carbonyloxy) amidine compounds as potent inhibitors against hepatitis C virus replication (2012)
  • Author(s): Kusano-Kitazume A, Sakamoto N, Okuno Y, Sekine-Osajima Y, Nakagawa M, Kakinuma S, Kiyonashi K, Nitta S, Murakawa M, Azuma S, Nishimura-Sakurai Y, Hagiwara M, Watanabe M
  • Journal Title: Antimicrob Agent Chemother Volume: 56(3) Pages: 1315-1323
• [Journal Article] Inhibition of hepatitis C virus NS5B polymerase by S-trityl-L-cysteine derivatives (2012)
  • Author(s): Kaushik-Basu N, Sakamoto N
  • Journal Title: Eur J Med Chem Volume: 49 Pages: 191-199
  • DOI: 10.1016/j.ejmech.2012.01.010
  • Peer Reviewed
• [Journal Article] Age and total ribavirin dose are independent predictors of relapse after interferon therapy in chronic hepatitis C revealed by data mining analysis (2012)
  • Author(s): Kurosaki M, Sakamoto N, Nakagawa M, et al.
  • Journal Title: Antivir Ther Volume: 17(1) Pages: 35-43
  • Peer Reviewed
• [Journal Article] Data mining model using simple and readily available factors could identify patients at high risk for hepatocellular carcinoma in chronic hepatitis C (2012)
  • Author(s): Kurosaki M, Sakamoto N, Nakagawa M, et al.
  • Journal Title: J Hepatol Volume: 56(3) Pages: 602-608
  • DOI: 10.1016/j.jhep.2011.09.011
  • Peer Reviewed
• [Journal Article] Use of human hepatocyte-like cells derived from induced pluripotent stem cells as a model for hepatocytes in hepatitis C virus infection (2011)
  • Author(s): Yoshida T, Takayama K, Kondoh M, Sakurai F, Tani H, Sakamoto N, Matsuura Y, Mizuguchi H, Yagi K
  • Journal Title: Biochem Biophys Res Commun Volume: 416(1-2) Pages: 119-124
• [Journal Article] Analysis of interferon signaling by infectious hepatitis C virus clones with substitutions of core amino acids 70 and 91 (2011)
  • Author(s): Funaoka Y, Sakamoto N, Nakagawa M, Kakinuma S, Suda S, Watanabe T, Nitta S, Kitazume A, Kiyohashi K, Murakawa M, Azuma S, Tsuchiya K, and Watanabe M
  • Journal Title: J Virol Volume: 85(12) Pages: 5986-5994
• [Journal Article] Studies on virus kinetics using infectious fluorescence-tagged hepatitis C virus cell culture (2011)
  • Author(s): Yamamoto M, Sakamoto N, Nakamura T, Itsui Y, Nakagawa M, Nishimura-Sakurai Y, Kakinuma S, Azuma S, Tsuchiya K, Kato T, Wakita T, Watanabe M
  • Journal Title: Hepatol Res Volume: 41 Pages: 258-269
• [Journal Article] Association between lipid accumulation and the cannabinoid system in Huh7 cells expressing HCV genes (2011)
  • Author(s): Toyoda M, Kitaoka A, Machida K, Nishinakagawa T, Yada R, Kohjima M, Kato M, Kotoh K, Sakamoto N, Shiota G, Nakamuta M, Nakashima M, Enjoji M
  • Journal Title: Int J Mol Med Volume: 27 Pages: 619-624
• [Journal Article] Association of gene expression involving innate immunity and genetic variation in IL28B with antiviral response (2011)
  • Author(s): Asahina Y, Sakamoto N, et al.
  • Journal Title: Hepatology Volume: 55(1) Pages: 20-29
  • DOI: 10.1 002/hep.24623
  • Peer Reviewed
• [Journal Article] Analysis of the complete open reading frame of hepatitis C virus in genotype 2a infection reveals critical sites influencing the response to peginterferon and ribavirin therapy (2011)
  • Author(s): Kadokura M, Nakagawa M, Sakamoto N, Watanabe M, et al.
  • Journal Title: Hepatol Int Volume: 5(3) Pages: 789-799
  • DOI: 10.1007/s12072-011-9267-x
  • Peer Reviewed
• [Journal Article] A new method for induced fit docking (GENIUS) and its application to virtual screening of novel HCV NS3-4A protease inhibitors (2011)
  • Author(s): Takaya D, Sakamoto N, et al.
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 19(22) Pages: 6892-6905
  • DOI: 10.1016/j.bmc.2011.09.023
  • Peer Reviewed
• [Journal Article] Serum interleukin-6 levels correlate with resistance to treatment of chronic hepatitis C infection with pegylated-interferon-alpha2b plus ribavirin (2011)
  • Author(s): Ueyama M, Nakagawa M, Sakamoto N, et al.
  • Journal Title: Antivir Ther Volume: 16 Pages: 1081-1091
  • Peer Reviewed
• [Journal Article] Genome-wide association study identified ITPA/DDRGK1 variants reflecting thrombocytopenia in pegylated interferon and ribavirin therapy for chronic hepatitis C (2011)
  • Author(s): Tanaka Y, Sakamoto N, et al.
  • Journal Title: Human Molecular Genetics Volume: 20(17) Pages: 3507-3516
  • DOI: 10.1093/hmg/ddr249
  • Peer Reviewed
• [Journal Article] Pretreatment Prediction of Anemia Progression by Peginterferon Plus Ribavirin Therapy in Chronic Hepatitis C : Decision-Tree Analysis (2011)
  • Author(s): Hiramatsu N, Sakamoto N, et al.
  • Journal Title: J Gastroenterol Volume: 46(9) Pages: 1111-1119
  • DOI: 10.1007/s00535-011-0412-z
  • Peer Reviewed
• [Journal Article] Analysis of the complete open reading frame of genotype 2b hepatitis C virus in association with the response to peginterferon and ribavirin therapy (2011)
  • Author(s): Kadokura M, Nakagawa M, Sakamoto N, Watanabe M, et al.
  • Journal Title: PLoS One Volume: 6(9) Pages: e24514
  • DOI: 10.1371/journal.pone.0024514
  • Peer Reviewed
• [Journal Article] Pretreatment prediction of response to peginterferon plus ribavirin therapy in genotype 1 chronic hepatitis C using data mining analysis (2011)
  • Author(s): Kurosaki M, Sakamoto N, et al.
  • Journal Title: J Gastroenterol Volume: 46(3) Pages: 401-409
  • DOI: 10.1007/s00535-010-0322-5
  • Peer Reviewed
• [Journal Article] Adenovirus vector-mediated assay system for hepatitis C virus replication (2011)
  • Author(s): Yoshida T, Sakamoto N, et al.
  • Journal Title: Nucleic Acids Res Volume: 39(10) Pages: e64
  • DOI: 10.1093/nar/gkrO47
  • Peer Reviewed
• [Journal Article] Inhibitory effect of a triterpenoid compound, with or without alpha interferon, on hepatitis C virus infection (2011)
  • Author(s): Watanabe T, Sakamoto N, Nakagawa M, Azuma S, Watanabe M, et al.
  • Journal Title: Antimicrob Agents Chemother Volume: 55(6) Pages: 2537-2545
  • DOI: 10.1128/AAC.01780-10
  • Peer Reviewed
• [Journal Article] Association of IL28B variants with response to pegylated-interferon alpha plus ribavirin combination therapy reveals intersubgenotypic differences between genotypes 2a and 2b (2011)
  • Author(s): Sakamoto N, Nakagawa M, Azuma S, Watanabe M, et al.
  • Journal Title: J Med Virol Volume: 83 Pages: 871-878
  • DOI: 10.1002/jmv.22038
  • Peer Reviewed
• [Journal Article] Cholestatic liver fibrosis and toxin-induced fibrosis are exacerbated in matrix metalloproteinase-2 deficient mice (2011)
  • Author(s): Onozuka I, Sakamoto N, Nakagawa M, Watanabe M, et al.
  • Journal Title: Biochem Biophys Res Commun Volume: 406 Pages: 134-140
  • DOI: 10.1016/j.bbrc.2011.02.012
  • Peer Reviewed
• [Journal Article] Pre-treatment prediction of response to pegylated-interferon plus ribavirin for chronic hepatitis C using genetic polymorphism in IL28B and viral factors (2011)
  • Author(s): Kurosaki M, Sakamoto N, Nakagawa M, Watanabe M, et al.
  • Journal Title: J Hepatol Volume: 54 Pages: 439-448
  • DOI: 10.1016/j.jhep.2010.07.037
  • Peer Reviewed
• [Journal Article] Sequences in the interferon sensitivity-determining region and core region of hepatitis C virus impact pretreatment prediction of response to PEG-interferon plus ribavirin : data mining analysis (2011)
  • Author(s): Kurosaki M, Sakamoto N, Nakagawa M, et al.
  • Journal Title: J Med Virol Volume: 83 Pages: 445-452
  • DOI: 10.1002/jmv.22005
  • Peer Reviewed
• [Presentation] ITPA gene variant protects against treatment-induced anemia and improves viral clearance by pegylated interferon-alfa and ribavirin therapy in chronic henatitis C patients (2011)
  • Author(s): Sakamoto N, Nakagawa M, Azuma S, Watanabe M, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] MMP-2 and MMP-14 derived from donor cells enhance therapeutic efficacy of liver cell transplantation in mice (2011)
  • Author(s): Kakinuma S, Sakamoto N, Watanabe M, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] A high-content screening assay using infectious fluorescence-tagged hepatitis C virus reveals candidates for small molecule inhibitors of viral entry (2011)
  • Author(s): Kusano-Kitazume A, Sakamoto N, Nakagawa M, Watanabe M, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] Loww of Wnt5A promotes biliary differentiation of murine hepatic stem/progenitor cells (2011)
  • Author(s): Kiyohashi K, Sakamoto N, Watanabe M, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] Antiviral effect of a novel interferon-inducible protein, IFI-27, against hepatitis C virus replication (2011)
  • Author(s): Itsuil Y, Sakamoto N, Watanabe M, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] IL-6-mediated intersubgenotypic variation of interferon sensitivity in hepatitis C virus genotype 2a/2b chimeric clones (2011)
  • Author(s): Suda G, Sakamoto N, Nakagawa M, Azuma S, Watanabe M, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] Prediction model of ribavirin-induced anemia incorporating ITPA genotype could identify chronic hepatitis C patients at high risk of relapse among virological responders to pegylated-interferon and ribavirin (2011)
  • Author(s): Kurosaki M, Sakamoto N, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] IL28B and ITPA gene variants correlate with treatment efficacy in pegylated-interferon plus ribavirin therapy for chronic hepatitis C (2011)
  • Author(s): Matsuura K, Sakamoto N, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] Mutations in the interferon sensitivity determining region of HCV, age and total ribavirin dose is an independent predictor of relapse among early virological responders to peg-interferon plus ribavirin therapy (2011)
  • Author(s): Kurosaki M, Sakamoto N, Nakagawa M, et al.
  • Organizer: 62th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: San Francisco
  • Year and Date: 2011-11-04
• [Presentation] Development of an RNA polymerase I-driven adenoviral vector and its application in an HCV replication assay (2011)
  • Author(s): Yoshida T, Sakamoto N, et al.
  • Organizer: 18th. International Meeting on Hepatitis C Virus & Related Viruses
  • Place of Presentation: Seattle
  • Year and Date: 2011-09-08
• [Presentation] Analysis of interferon signaling by infectious hepatitis C virus clones with substitutions of core amino acids 70 and 91 (2011)
  • Author(s): Sakamoto N, Nakagawa M, Watanabe M, et al.
  • Organizer: 18th. International Meeting on Hepatitis C Virus & Related Viruses
  • Place of Presentation: Seattle
  • Year and Date: 2011-09-08
• [Presentation] HCV-NS4B targets STING and abrogates RIG-I-mediated type-I interferon-dependent innate immune response (2011)
  • Author(s): Nitta S, Sakamoto N, Nakagawa M, Watanabe M, et al.
  • Organizer: 18th. International Meeting on Hepatitis C Virus & Related Viruses
  • Place of Presentation: Seattle
  • Year and Date: 2011-09-08
• [Presentation] Development of RNA pol-driven adenovirus vector expressing hepatitis C virus replicon (2011)
  • Author(s): Yamagishi Y, Sakamoto N, et al.
  • Organizer: Experimental Biology 2011
  • Place of Presentation: Washington
  • Year and Date: 2011-04-13
• [Patent(Industrial Property Rights)] C型肝炎ウイルスの増殖を抑制する医薬組成物 (2011)
  • Inventor(s): 坂本直哉、渡辺守、北詰晶子、萩原正敏、奥野友紀子
  • Industrial Property Rights Holder: 東京医科歯科大学
  • Industrial Property Number: 特許、特願2011-194082
  • Filing Date: 2011-09-06