Establishment of in vitro selection system by multi anti-HIV drugs

• Project/Area Number: 24590199
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Medical pharmacy
• Research Institution: National Institute of Infectious Diseases (2012, 2014); The University of Tokyo (2013)
• Principal Investigator: HARADA Shigeyoshi 国立感染症研究所, その他部局等, 主任研究官 (30508643)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: HIV / AIDS / 抗HIV療法 / 薬剤耐性 / 耐性誘導 / 臨床分離株

Outline of Final Research Achievements

The aim of study was to develop an in vitro selection system using human immunodeficiency virus type 1 (HIV-1) by multi anti-HIV drugs. In this study, we determined that (i) the HIV integrase inhibitor raltegravir (RAL)-resistant clone became highly sensitive to the CCR5 antagonist maraviroc (MVC) and (ii) MVC-resistant clone became highly sensitive to the anti-envelope (Env) neutralizing antibodies, respectively.
Furthermore, using established this system, it was determined that acquisitions of resistance mutations were significantly decreased during in vitro selection by multi anti-HIV drugs rather than only single drugs. Thus, this novel system is indicated a useful tool for investigating the compatibility of combined anti-retroviral therapy.

Research Products

• [Journal Article] Impact of maraviroc-resistant and low-CCR5- adapted mutations induced by in vitro passage on sensitivity to anti-envelope neutralizing antibodies. (2014)
  • Author(s): Kazuhisa Yoshimura, Shigeyoshi Harada, Samatchaya Boonchawalit, Yoko Kawanami, Shuzo Matsushita.
  • Journal Title: Journal of General Virology Volume: 95 Issue: 8 Pages: 1816-1826
  • DOI: 10.1099/vir.0.062885-0
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A CD4 Mimic as an HIV Entry Inhibitors: Pharmacokinetics (2013)
  • Author(s): Hashimoto, C., Narumi, T., Otsuki, H., Hirota, Y., Arai, H., Yoshimura, K., Harada, S., Ohashi, N., Nomura, W., Miura, T., Igarashi, T., Matsuhita, S., Tamamura, H.
  • Journal Title: Bioorg. Med. Chem. Volume: 21 Issue: 24 Pages: 7884-7889
  • DOI: 10.1016/j.bmc.2013.10.005
  • Peer Reviewed
• [Journal Article] CD4 Mimics as HIV Entry Inhibitors: Lead Optimization Studies of the Aromatic Substituents. (2013)
  • Author(s): Narumi, T., Arai, H., Yoshimura, K., Harada, S., Hirota, Y., Ohashi, N., Hashimoto, C.. Nomura, W., Matsushita, S. and Tamamura, H.
  • Journal Title: Bioorg. Med. Chem. Volume: 8 Issue: 9 Pages: 2518-2526
  • DOI: 10.1016/j.bmc.2013.02.041
  • Peer Reviewed
• [Journal Article] Impact of antiretroviral pressure on selection of primary human immunodeficiency virus type 1 envelope sequences in vitro. (2013)
  • Author(s): Shigeyoshi Harada, Kazuhisa Yoshimura, Aki Yamaguchi, Samatchaya Boonchawalit, Keisuke Yusa, Shuzo Matsushita.
  • Journal Title: Journal of General Virology Volume: 94 Issue: 5 Pages: 933-943
  • DOI: 10.1099/vir.0.047167-0
  • Peer Reviewed
• [Presentation] Mutations at the bottom of the Phe43 cavity are responsible for cross-resistance to NBD analogues. (2015)
  • Author(s): Shigeyoshi Harada, Yu Irahara, Samatchaya Boonchawalit, Mai Goryo, Hirokazu Tamamura, Tetsuro Matano, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: 22th Conference on Retroviruses and Opportunistic Infections (CROI 2015)
  • Place of Presentation: Seattle, WA, U.S.A.
  • Year and Date: 2015-02-23 – 2015-02-26
• [Presentation] CD4類似低分子化合物 (CD4MCs) とgp120. (2014)
  • Author(s): 原田恵嘉
  • Organizer: 第28回 日本エイズ学会学術集会・総会
  • Place of Presentation: 大阪
  • Year and Date: 2014-12-03 – 2014-12-05
• [Presentation] CD4類似低分子化合物誘導体 (CD4MCs) の耐性プロファイルと分子動力学的機構解析. (2014)
  • Author(s): 原田恵嘉, 横山勝, Samatchaya Boonchawalit, 佐藤裕徳, 松下修三, 吉村和久.
  • Organizer: 第62回 日本ウイルス学会学術集会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-10 – 2014-11-12
• [Presentation] Resistance Profile of CD4 Mimic Small Compounds (CD4MCs) and the Structure Analysis by Molecular Dynamic (MD) Simulation. (2014)
  • Author(s): Shigeyoshi Harada, Masaru Yokoyama, Samatchaya Boonchawalit, Hironori Sato, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: HIV Research for Prevention 2014
  • Place of Presentation: Cape Town, South Africa
  • Year and Date: 2014-10-28 – 2014-10-31
• [Presentation] Impact of maraviroc (MVC)-resistant mutations in the C1 and C4 regions of gp120 on sensitivity to antibody-mediated neutralization. (2014)
  • Author(s): Samatchaya Boonchawalit, Shigeyoshi Harada, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: 15th Kumamoto AIDS Seminar
  • Place of Presentation: Kumamoto, Japan
  • Year and Date: 2014-10-01 – 2014-10-03
• [Presentation] Impact of Maraviroc (MVC)-resistant mutations in C1 and C4 regions of gp120 on sensitivity to antibody-mediated neutralization. (2014)
  • Author(s): amatchaya Boonchawalit, Shigeyoshi Harada, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: 20th International AIDS Conference Melbourne
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 2014-07-20 – 2014-07-25
• [Presentation] 新規エントリー阻害剤の組み合わせによる抗ウイルス効果と耐性変異の解析. (2014)
  • Author(s): 原田恵嘉.
  • Organizer: 第17回 日本レトロウイルス研究会 夏期セミナー
  • Place of Presentation: 静岡
  • Year and Date: 2014-07-03 – 2014-07-05
• [Presentation] 新規エントリー阻害剤の組み合わせによる抗ウイルス効果と耐性変異の解析. (2014)
  • Author(s): 原田恵嘉, 横山勝, Samatchaya Boonchawalit, 佐藤裕徳, 松下修三, 吉村和久.
  • Organizer: 第16回 白馬シンポジウム
  • Place of Presentation: 熊本
  • Year and Date: 2014-06-13 – 2014-06-14
• [Presentation] Impact of CD4 mimetics-resistant mutations on susceptibilities to anti-Env nMAbs. (2014)
  • Author(s): Shigeyoshi Harada, Samatchaya Boonchawalit, Tetsuo Narumi, Hirokazu Tamamura, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: 21th Conference on Retroviruses and Opportunistic Infections (CROI 2014)
  • Place of Presentation: Boston, MA, U.S.A.
• [Presentation] バルクおよびクローンウイルスを用いたCD4類似低分子化合物誘導体に対するin vitro 耐性ウイルス誘導. (2013)
  • Author(s): 原田恵嘉, 鳴海哲夫, Samatchaya Boonchawalit, 玉村啓和, 松下修三, 吉村和久.
  • Organizer: 第27回 日本エイズ学会学術集会・総会
  • Place of Presentation: 熊本
• [Presentation] 抗V3抗体および低分子CD4ミミック曝露後投与によるアカゲザルでのSHIV複製抑制. (2013)
  • Author(s): 大附寛幸,丸田泰広,橋本智恵,鳴海哲夫,廣田雄樹, 原田恵嘉, 三浦智行, 吉村和久,玉村啓和,松下修三, 五十嵐樹彦.
  • Organizer: 第27回 日本エイズ学会学術集会・総会
  • Place of Presentation: 熊本
• [Presentation] Analysis of relationships between Maraviroc (MVC) resistant mutations and sensitivity to antibody-mediated neutralization. (2013)
  • Author(s): Samatchaya Boonchawalit, 原田恵嘉, 松下修三, 吉村和久.
  • Organizer: 第27回 日本エイズ学会学術集会・総会
  • Place of Presentation: 熊本
• [Presentation] 細胞内ドメイン欠損Envを有するHV/SIV粒子の作製. (2013)
  • Author(s): 五領舞衣, 原田恵嘉, 石井洋, 吉村和久, 俣野哲朗.
  • Organizer: 第27回 日本エイズ学会学術集会・総会
  • Place of Presentation: 熊本
• [Presentation] インテグラーゼ阻害剤ラルテグラビルがMVC耐性HIV-1 Env領域に与える影響. (2013)
  • Author(s): 原田恵嘉, Samatchaya Boonchawalit, 松下修三, 吉村和久.
  • Organizer: 第61回 日本ウイルス学会学術集会
  • Place of Presentation: 神戸
• [Presentation] In vitro selection of bifunctional CD4 mimic small compounds (NBD analogues) using bulk and cloned primary isolates. (2013)
  • Author(s): Shigeyoshi Harada, Samatchaya Boonchawalit, Tetsuo Narumi, Hirokazu Tamamura, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: 14th Kumamoto AIDS Seminar, Kumamoto
  • Place of Presentation: Kumamoto, Japan
• [Presentation] Analysis of relationships between Maraviroc (MVC) resistant mutations and sensitivity to antibody-mediated neutralization. (2013)
  • Author(s): Samatchaya Boonchawalit, Shigeyoshi Harada, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: 14th Kumamoto AIDS Seminar, Kumamoto
  • Place of Presentation: Kumamoto, Japan
• [Presentation] Analysis of interaction between gp120 and CD4 mimic small compounds that enhance the activity of anti-HIV neutralizing antibodies. (2013)
  • Author(s): Shigeyoshi Harada, Samatchaya Boonchawalit, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: AIDS Vaccine 2013
  • Place of Presentation: Barcelona, Spain
• [Presentation] Characterization of highly neutralizing antibody sensitive HIV-1 gp120 induced under high concentrations of maraviroc (MVC) in vitro. AIDS Vaccine 2013 (2013)
  • Author(s): Samatchaya Boonchawalit, Shigeyoshi Harada, Shuzo Matsushita, Kazuhisa Yoshimura.
  • Organizer: AIDS Vaccine 2013
  • Place of Presentation: Barcelona, Spain
• [Presentation] In vitro selection of bifunctional CD4 mimic small compounds (NBD analogues) using bulk and cloned primary isolates. (2013)
  • Author(s): 原田恵嘉, Samatchaya Boonchawalit, 松下修三, 吉村和久.
  • Organizer: 第15回 白馬シンポジウム
  • Place of Presentation: 名古屋
• [Presentation] Impact od deletion in cytoplasmic tail on Env incorporation into HIV/SIV particles. (2013)
  • Author(s): 五領舞衣, 原田恵嘉, 石井洋,俣野哲朗, 吉村和久.
  • Organizer: 第15回 白馬シンポジウム
  • Place of Presentation: 名古屋
• [Presentation] バルクおよびクローンウイルスを用いたCD4類似低分子化合物の耐性誘導. (2013)
  • Author(s): 原田恵嘉.
  • Organizer: 第16回 日本レトロウイルス研究会 夏期セミナー
  • Place of Presentation: 静岡
• [Presentation] CD4 Mimic Small Compounds, NBD-556 and Its Analogues Bind at 3 Amino Acid Positions in the gp120 CD4 Cavity (2013)
  • Author(s): Shigeyoshi Harada, Samatchaya Boonchawalit, Tetsuo Narumi, Hirokazu Tamamura, Shuzo Matsushita, and Kazuhisa Yoshimura
  • Organizer: 20th Conference on Retroviruses and Opportunistic Infections (CROI)
  • Place of Presentation: Atlanta, GA, USA
• [Presentation] Impact of Maraviroc-resistant and Low CCR5-adapted Mutations Induced in vitro Passage on Sensitivity to Anti-Env Neutralizing Antibodies (2013)
  • Author(s): Kazuhisa Yoshimura, Shigeyoshi Harada, Samatchaya Boonchawalit, and Shuzo Matsushita
  • Organizer: 20th Conference on Retroviruses and Opportunistic Infections (CROI)
  • Place of Presentation: Atlanta, GA, USA
• [Presentation] R5臨床分離株を用いたCD4類似低分子化合物誘導体に対するin vitro 耐性ウイルス誘導 (2012)
  • Author(s): 原田恵嘉、鳴海哲夫、玉村啓和、松下修三、吉村和久
  • Organizer: 第26回日本エイズ学会学術集会・総会
  • Place of Presentation: 横浜市
• [Presentation] In vitro induction of ten CD4 mimic small compounds, NBD-556 and its analogues, resistant variants using primary R5 HIV-1. (2012)
  • Author(s): Shigeyoshi Harada, Hiroshi Arai, Tetsuo Narumi, Hirokazu Tamamura, Shuzo Matsushita, and Kazuhisa Yoshimura
  • Organizer: 19th International AIDS Conference.
  • Place of Presentation: Washington, D.C., U.S.A.
• [Presentation] CD4類似低分子化合物の耐性機序および結合部位の検討. (2012)
  • Author(s): 原田 恵嘉
  • Organizer: 第15回 SUMMER RETROVIRUS CONFERENCE
  • Place of Presentation: 京都市
• [Presentation] HIV-1におけるCD4類似低分子化合物に対する耐性機序および結合部位の検討 (2012)
  • Author(s): 原田 恵嘉
  • Organizer: 第9回ウイルス学キャンプ in 湯河原
  • Place of Presentation: 静岡県熱海市
• [Presentation] R5臨床分離株を用いた10種のCD4類似低分子化合物に対するin vitro 耐性ウイルス誘導 (2012)
  • Author(s): 原田 恵嘉
  • Organizer: 第14回白馬シンポジウム
  • Place of Presentation: 京都市