本邦で見出された新規多剤耐性変異N348IのHIV逆転写酵素に及ぼす影響

2009 Fiscal Year Annual Research Report

• Project/Area Number: 21590591
• Research Institution: Tohoku University
• Principal Investigator: 児玉 栄一 Tohoku University, 病院, 助教 (50271151)
• Keywords: ウイルス / HIV / 薬剤耐性 / 逆転写酵素 / 多剤耐性 / 臨床分離株

Research Abstract

本研究ではhuman immunodeficiency virus type1(HIV-1)逆転写酵素(RT)の酵素活性に直接関与していないと考えられてきたconnection subdomainに位置する新規多剤耐性変異N348Iの意義を解明することを目的としている。平成21年度はこのN348Iが他の耐性変異とどのような相互作用をするのか、検討するために感染性分子クローンHIVをsite directed mutagenesis法を用いて作製した。特に核酸系逆転写酵素阻害剤関連耐性変異を中心に作製し、293T細胞に遺伝子導入し感染性HIVが同収されることを確認した。
一方で他のグループからconnection subdomainの他の変異が多剤耐性に関与しているのではないかという報告が相次ぎ、研究代表者はN348Iと比較するために独自に本邦で薬剤治療歴のないHIV感染患者からsubtype Bを101株、non-B subtypeを16株分離し、connection subdomainにおける変異の導入程度を検討した。これまでの他のグループからの報告ではG312QやG335Dが関連するように報告されていたが、我々の解析では未治療者からもそのような変異を有するHIVが分離されること、特にG335Dはnon-B subtype HIVでは約70%程度に見られることから、薬剤によって誘導されたというよりはもともとのsubtype特異的なpolymorphismであると考えられた。さらに感染性分子クローンを用いた解析でN348I以外は多剤耐性を示さないことが分かり、おもに非核酸系逆転写酵素阻害剤であるネビラピンに対して耐性を未すことを明らかとした。現在本邦ではネビラピンではなくエファビレンツが主に使われているため、connection subdomainの変異としてはN348I以外はあまり留意する必要がないと考えられる。

Research Products

• [Journal Article] Synthesis and Biological Evaluation of Selective CXCR4 Antagonists Containing Alkene Dipeptide Isosteres. (2010)
  • Author(s): Tetsuo Narumi, Ryoko Hayashi, Kenji Tomita, Kazuya Kobayashi, Noriko Tanahara, Hiroaki Ohno, Takeshi Naito, Eiichi Kodama, Masao Matsuoka, Shinya Oishi, Nobutaka Fujii.
  • Journal Title: Organic & Biomolecular Chemistry 8 Pages: 616-621
  • Peer Reviewed
• [Journal Article] Binding of Multivalent Anionic Porphyrins to V3 Loop Fragments of HIV-1 gp120 and Their Antiviral Activity. (2010)
  • Author(s): Kenji Watanabe, Shigeru Negi, Yukio Sugiura, Akiko Kiriyama, Akino Honbo, Katsumi Iga, Eiichi N.Kodama, Takeshi Naitoh, Masao Matsuoka, Koji Kano.
  • Journal Title: Chemistry-An Asian Journal 5 Pages: 825-834
  • Peer Reviewed
• [Journal Article] (2010)
  • Author(s): 児玉栄一, 他
  • Journal Title: 新規抗HIV薬の開発 特集・抗ウイルス薬物療法の現状と今後の展望(医薬ジャーナル) Pages: 99-102
• [Journal Article] Electrostatically constrained α-helical peptide inhibits replication of HIV-1 resistant to enfuvirtide. (2009)
  • Author(s): Hiroki Nishikawa, Shota Nakamura, Eiichi Kodama, Saori Ito, Keiko Kajiwara, Kazuki Izumi, Yasuko Sakagami, Shinya Oishi, Tadayasu Ohkubo, Yuji Kobayashi, Akira Otaka, Nobutaka Fujii, Masao Matsuoka
  • Journal Title: International Journal of Biochemistry and Cell Biology 41 Pages: 891-899
  • Peer Reviewed
• [Journal Article] Design of peptide-based inhibitors for HIV-1 strains resistant to T-20 (2009)
  • Author(s): Kazuki Izumi, Eiichi Kodama, Kazuya Shimura, Yasuko Sakagami, Kentaro Watanabe, Saori Ito, Tsuyoshi Watabe, Yukihiro Terakawa, Hiroki Nishikawa, Stefan G.Sarafianos, Kazuo Kitaura, Shinya Oishi, Nobutaka Fujii, Masao Matsuoka
  • Journal Title: Journal of Biological Chemistry 284 Pages: 4914-4920
  • Peer Reviewed
• [Journal Article] SC29EK, a peptide fusion inhibitor with enhanced α-helicity, inhibits replication of human immunodeficiency virus type 1 mutants resistant to enfuvirtide. (2009)
  • Author(s): akeshi Naito, Kazuki Izumi, Eiichi Kodama, Yasuko Sakagami, Keiko Kajiwara, Hiroki Nishikawa, Kentaro Watanabe, Stefan G.Sarafianos, Shinya Oishi, Nobutaka Fujii, Masao Matsuoka.
  • Journal Title: Antimicrobal Agents and Chemotherapy 53 Pages: 1013-1018
  • Peer Reviewed
• [Journal Article] Synonymous mutations in stem-loop III of Rev responsive elements enhance HIV-1 replication impaired by primary mutations for resistance to enfuvirtide. (2009)
  • Author(s): Mariko Ueno, Eiichi N.Kodama, Kazuya Shimura, Yasuteru Sakurai, Keiko Kajiwara, Yasuko Sakagami, Shinya Oishi, Nobutaka Fujii, Masao Matsuoka.
  • Journal Title: Antiviral Research 82 Pages: 67-72
  • Peer Reviewed
• [Journal Article] Clinical relevance of substitutions in the connection subdomain and R Nase H domains of HIV-1 reverse transcriptase from a cohort of antiretroviral treatment-naive patients. (2009)
  • Author(s): Atsuko Hachiya, Kazuki Shimane, Stefan G.Sarafianos, Eiichi N.Kodama, Yasuko Sakagami, Fujie Negishi, Hirokazu Koizumi, Hiroyuki Gatanaga, Masao Matsuoka, Masafumi Takiguchi, Shinichi Oka.
  • Journal Title: Antiviral Research 82 Pages: 115-121
  • Peer Reviewed
• [Journal Article] Peptide bond mimicry by (E)-alkene and (Z)-fluoroalkene peptide isosteres : synthesis and bioevaluation of alpha-helical anti-HIV peptide analogues. (2009)
  • Author(s): Shinya Oishi, Hirotaka Kamitani, Yasuyo Kodera, Kentaro Watanabe, Kazuya Kobayashi, Tetsuo Narumi, Kenji Tomita, Hiroaki Ohno, Takeshi Naito, Eiichi Kodama, Masao Matsuoka, Nobutaka Fujii
  • Journal Title: Organic & Biomolecular Chemistry 7 Pages: 2872-2877
  • Peer Reviewed
• [Journal Article] Design and synthesis of membrane fusion inhibitors against the feline immunodeficiency virus. (2009)
  • Author(s): Shinya Oishi, Yasuyo Kodera, Hiroki Nishikawa, Kamitani H, Tsuyoshi Watabe, Hiroaki Ohno, Tadafumi Tochikura, Kazuki Shimane, Eiichi Kodama, Masao Matsuoka, Fuminori Mizukoshi, Hajime Tsujimoto H, Nobutaka Fujii
  • Journal Title: Bioorganic & Medicinal Chemistry 17 Pages: 4916-4920
  • Peer Reviewed
• [Journal Article] X-ray Crystallographic Study of an HIV-1 Fusion Inhibitor with the gp41 S138A Substitution. (2009)
  • Author(s): Tsuyoshi Watabe, Yukihiro Terakawa, Kentaro Watanabe, Hiroaki Ohno, Hiroaki Nakano, Toru Nakatsu, Hiroaki Kato, Kazuki Izumi, Eiichi Kodama, Masao Matsuoka, Kazuo Kitaura, Shinya Oishi, Nobutaka Fujii.
  • Journal Title: Journal of Molecular Biology 392 Pages: 657-665
  • Peer Reviewed
• [Journal Article] Bioorganic Synthesis of End-capped Anti-HIV Peptides by Simultaneous Cyanocysteine-mediated Cleavages of Recombinant Proteins. (2009)
  • Author(s): Michinori Tanaka, Kazumi Kajiwara, Rei Tokiwa, Kentaro Watanabe, Hiroaki Ohno, Hiroko Tsutsumi, Yoji Hata, Kazuki Izumi, Eiichi Kodama, Masao Matsuoka, Shinya Oishi, Nobutaka Fujii
  • Journal Title: Bioorganic & Medicinal Chemistry 17 Pages: 7487-7492
  • Peer Reviewed
• [Journal Article] Bioorganic Synthesis of a Recombinant HIV-1 Fusion Inhibitor, SC35EK, with an N-Terminal Pyroglutamate Capping Group. (2009)
  • Author(s): Kazumi Kajiwara, Kentaro Watanabe, Rei Tokiwa, Tomoko Kurose, Hiroaki Ohno, Hiroko Tsutsumi, Yoji Hata, Kazuki Izumi, Eiichi Kodama, Masao Matsuoka, Shinya Oishi, Nobutaka Fujii.
  • Journal Title: Bioorganic & Medicinal Chemistry 17 Pages: 7964-7970
  • Peer Reviewed
• [Journal Article] Mechanism of inhibition of HIV-1 reverse transcriptase by 4'-ethynyl-2-fluoro-2'-doxyadenosine triphosphate, a translocation defective reverse transcriptase inhibitor. (2009)
  • Author(s): Eleftherios Michailidis, Bruno Marchand, Eiichi N.Kodama, Kamlendra Singh, Masao Matsuoka, Karen A.Kirby, Emily M.Ryan, Ali M.Sawani, Eva Nagy, Noriyuki Ashida, Hiroaki Mitsuya, Michael A.Parniak, Stefan G.Sarafianos.
  • Journal Title: Journal of Biological Chemistry 284 Pages: 35681-35691
  • Peer Reviewed
• [Journal Article] (2009)
  • Author(s): 児玉栄一、服部俊夫, 他
  • Journal Title: HIV感染症と増殖のメカニズム-抗ウイルス剤の作用点を中心に- 特集HIV感染症 流行の現状と最新の治療(日本内科学会雑誌) Pages: 2754-2761
• [Presentation] Biochemical characterization of the HIV reverse transcriptase connection domain multi-class drug resistance mutation N348I. (2009)
  • Author(s): Matthew Schuckmann, Atsuko Hachiya, Ei-ichi N.Kodama, Bruno Marchand, Eleftherios Michailidis, Kamlendra Singh, Yasuko Sakagami, Shinichi Oka, Stefan G.Sarafianos.
  • Organizer: 10th Annual Symposium on Antiviral Drug Resistance
  • Place of Presentation: アメリカ、リッチモンド
  • Year and Date: 2009-11-16
• [Presentation] The NNRTI-resistant Y181C HIV-1 reverse transcriptase is hyper-susceptible to 4'-ethynyl-2-fluoro deoxyadenosine(EFDA) (2009)
  • Author(s): Yee Tsuey Ong, Eleftherios Michailidis, Bruno Marchand, Matthew S.Schuckmann, Kamlendra Singh, Ei-Ichi Kodama, Masao Matsuoka, Hiroaki Mitsuya, Eva Nagy, Michael A.Parniak, Stefan G.Sarafianos.
  • Organizer: 10th Annual Symposium on Antiviral Drug Resistance
  • Place of Presentation: アメリカ、リッチモンド
  • Year and Date: 2009-11-16
• [Presentation] K70Q expands multi-drug resistance (MDR) of HIV-1 containing "Q151M complex" reverse transcriptase mutations. (2009)
  • Author(s): Atsuko Hachiya, Eiichi N.Kodama, Matthew M.Schuckmann, Karen A.Kirby, Eleftherios Michailidis, Yasuko Sakagami, Kamlendra Singh, Shinichi Oka, Stefan G.Sarafianos.
  • Organizer: 10th Annual Symposium on Antiviral Drug Resistance
  • Place of Presentation: アメリカ、リッチモンド
  • Year and Date: 2009-11-16
• [Presentation] Parniak 4'-Ethynyl-2-fluoro-2'-deoxyadenosine(EFDA), a translocation defective reverse transcriptase inhibitor with highly potent activity against diverse multi-drug-resistant HIV-1 variants. (2009)
  • Author(s): Hiroaki Mitsuya, Stefan G.Sarafianos, Eiichi Kodama, Hirotomo Nakata, Seiji Okada, Michael A.
  • Organizer: 10th Annual Symposium on Antiviral Drug Resistance
  • Place of Presentation: アメリカ、リッチモンド
  • Year and Date: 2009-11-15
• [Presentation] Drug design for resistant HIV (2009)
  • Author(s): Kodama E.
  • Organizer: 10th Kumamoto AIDS seminar/AIDS GCOE International Symposium Satellite symposium
  • Place of Presentation: 熊本(招待講演)
  • Year and Date: 2009-09-30