Structural analysis of a reverse transcriptase inhibitor, EFdA, potent against drug-resistant HIV

• Project/Area Number: 16K08826
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Virology
• Research Institution: National Center for Global Health and Medicine
• Principal Investigator: MAEDA KENJI 国立研究開発法人国立国際医療研究センター, その他部局等, 室長 (50758323)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: HIV / 逆転写酵素阻害剤 / 薬剤耐性 / 構造解析 / 新薬開発 / EFdA/MK-8591 / 逆転写酵素阻害剤（NRTI） / 逆転写酵素 / 抗ウイルス療法 / ウイルス / 感染症 / 微生物

Outline of Final Research Achievements

EFdA/MK- 8591, a nucleoside reverse transcriptase inhibitor (NRTI), is a potent and promising long-acting anti-HIV-1 agent. EFdA and its derivatives possess a modified 4’-moiety and potently inhibit the replication of HIV-1 strains resistant to existing NRTIs. Here, we report that EFdA and NRTIs with a 4’-ethynyl-moiety exerted activity against HIV-1 with an M184V mutation and multiple NRTI-resistant HIV-1s, whereas NRTIs with other moieties (e.g., 4'-methyl) did not show this activity. Structural analysis indicated that EFdA and 4’-ethynyl-NRTIs (but not other 4’-modified NRTIs), formed strong van der Waals interactions with critical amino acid residues of reverse transcriptase. Such interactions were maintained even in the presence of a broad resistance-endowing M184V substitution, thus potently inhibiting drug-resistant HIV-1 strains. These findings also explain the mechanism for the potency of EFdA and provide insights for further design of anti-HIV-1 therapeutics.

Academic Significance and Societal Importance of the Research Achievements

本研究で行われた強力な逆転写酵素阻害剤（EFdA）の薬剤耐性メカニズムに対する構造学的研究から得られた知見は薬剤耐性HIVに対して有効な新規抗HIV薬の設計・開発に寄与するものであり、構造学的エビデンスに基づいた効率的な薬剤開発の有効性を示唆するものである。
さらに本研究で得られた阻害剤のHIV薬剤耐性に関わる構造についてはHIV逆転写酵素（HIV-RT）とB型肝炎ウイルスのHBV-RTに対する阻害剤の親和性の違いにも関連していることが分かってきており、今後各種のRT（およびDNAポリメラーゼ）とその阻害剤に関する構造・活性比較解析研究の進展と新たな治療薬開発が期待される。

Research Products

• [Journal Article] Active-site deformation in the structure of HIV-1 RT with HBV-associated septuple amino acid substitutions rationalizes the differential susceptibility of HIV-1 and HBV against 4?-modified nucleoside RT inhibitors (2019)
  • Author(s): Yasutake Yoshiaki、Hattori Shin-ichiro、Tamura Noriko、Matsuda Kouki、Kohgo Satoru、Maeda Kenji、Mitsuya Hiroaki
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 509 Issue: 4 Pages: 943-948
  • DOI: 10.1016/j.bbrc.2019.01.026
  • Peer Reviewed / Open Access
• [Journal Article] The High Genetic Barrier of EFdA/MK-8591 Stems from Strong Interactions with the Active Site of Drug-Resistant HIV-1 Reverse Transcriptase (2018)
  • Author(s): Takamatsu Yuki、Das Debananda、Kohgo Satoru、Hayashi Hironori、Delino Nicole S.、Sarafianos Stefan G.、Mitsuya Hiroaki、Maeda Kenji
  • Journal Title: Cell Chemical Biology Volume: 25 Issue: 10 Pages: 1268-1278
  • DOI: 10.1016/j.chembiol.2018.07.014
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Synthesis of 4’‐Substituted Purine 2’‐Deoxynucleosides and Their Activity against Human Immunodeficiency Virus Type 1 and Hepatitis B Virus. (2018)
  • Author(s): 3.Satoru Kohgo, Shuhei Imoto, Ryoh Tokuda, Yuki Takamatsu, Nobuyo Higashi-Kuwata, Manabu Aoki, Masayuki Amano, Hisao Kansui, Kengo Onitsuka, Kenji Maeda, Hiroaki Mitsuya.
  • Journal Title: ChemistrySelect. Volume: 3(11) Issue: 11 Pages: 3313-3317
  • DOI: 10.1002/slct.201800527
  • Peer Reviewed / Open Access
• [Journal Article] HIV-1 with HBV-associated Q151M substitution in RT becomes highly susceptible to entecavir: structural insights into HBV-RT inhibition by entecavir. (2018)
  • Author(s): Yasutake Y, Hattori SI, Hayashi H, Matsuda K, Tamura N, Kohgo S, Maeda K, Mitsuya H.
  • Journal Title: Sci Rep. Volume: 8(1) Issue: 1 Pages: 1624-1624
  • DOI: 10.1038/s41598-018-19602-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Synthesis of 4’-Substituted Purine 2’-Deoxyucleosides and Their Activity against Human Immunodeficiency Virus and Hepatitis B Virus. (2018)
  • Author(s): Kohgo S, Imoto S, Tokuda R, Takamatsu Y, Higashi-Kuwata N, Aoki M, Amano M, Kansui H, Onitsuka K, Maeda K, and Mitsuya H
  • Journal Title: ChemistrySelect Volume: in press
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Restoration of immune surface molecules in Kaposi sarcoma-associated herpes virus infected cells by lenalidomide and pomalidomide (2017)
  • Author(s): Davis David A.、Mishra Suraj、Anagho Holda A.、Aisabor Ashley I.、Shrestha Prabha、Wang Victoria、Takamatsu Yuki、Maeda Kenji、Mitsuya Hiroaki、Zeldis Jerome B.、Yarchoan Robert
  • Journal Title: Oncotarget Volume: 8 Issue: 31 Pages: 50342-50358
  • DOI: 10.18632/oncotarget.17960
  • Open Access
• [Presentation] 新規抗HBV化合物CFCPの発見とその抗HBV活性及び薬理学的動態の解析 (2018)
  • Author(s): 鍬田伸好、青木宏美、林佐奈衣、熊本浩樹、服部真一朗、前田賢次、田中靖人、満屋裕明
  • Organizer: 第28回抗ウイルス療法学会学術集会・総会
• [Presentation] 4’-Modified NRTIs’ Potent Anti-HIV Activity Stems from Strong RT Active Site Binding. (2017)
  • Author(s): Takamatsu Y, Das D, Kohgo S, Hattori S, Hayashi H, Matsuda K, Stefan G. Sarafianos SG, Hiroaki Mitsuya H, and Maeda K.
  • Organizer: 2017 Conference on Retroviruses and Opportunistic Infections (CROI)
  • Place of Presentation: Seattle WA
  • Year and Date: 2017-02-12
  • Int'l Joint Research
• [Presentation] HIV潜伏感染細胞を標的とした新規治療薬開発に有効なin vitro評価系の開発 (2017)
  • Author(s): 1.松田 幸樹、服部 真一朗、土屋 亮人、小早川 拓也、大橋 南美、野村 渉、原田 恵嘉、佐藤 賢文、吉村 和久、玉村 啓和、満屋 裕明、前田 賢次
  • Organizer: 第31回日本エイズ学会学術集会・総会
• [Presentation] Analysis of molecular mechanism of HIV-1 latent infection & reactivation and development of novel therapeutics active against latent HIV-1 infection. (2017)
  • Author(s): 3.Kouki Matsuda, Shin-ichiro Hattori, Kiyoto Tsuchiya, Takuya Kobayakawa, Nami Ohashi, Wataru Nomura, Shigeyoshi Harada, Yorifumi Satou, Kazuhisa Yoshimura, Hirokazu Tamamura, Hiroaki Mitsuya, and Kenji Maeda
  • Organizer: 第65回日本ウイルス学会学術集会
• [Presentation] Molecular simulation of EFdA and related analogs to determine the structural basis of HIV-1 drug resistance (2017)
  • Author(s): 8.Debananda Das, Yuki Takamatsu, Satoru Kohgo, Shinichiro Hattori, Hironori Hayashi, Kouki Matsuda, Stefan G. Sarafianos, Hiroaki Mitsuya, and Kenji Maeda
  • Organizer: ACS' 253rd National Meeting & Exposition
  • Int'l Joint Research
• [Presentation] 逆転写酵素阻害剤EFdA(MK-8591)に対する薬剤耐性機序の構造学的解析と耐性株に有効な新規薬剤の設計・開発 (2016)
  • Author(s): 前田賢次、高松悠樹、向後悟、服部真一朗、鍬田伸好、林宏典、Debananda Das、満屋裕明.
  • Organizer: 第30回日本エイズ学会学術集会・総会.
  • Place of Presentation: 鹿児島
  • Year and Date: 2016-11-24
• [Presentation] Progress in the therapy for HIV/AIDS: development of molecular-targeting approach for antiretroviral drugs. (2016)
  • Author(s): Maeda K, and Mitsuya H.
  • Organizer: 2nd Kumamoto IRCMS International Symposium and 17th Kumamoto AIDS Seminar
  • Place of Presentation: 熊本
  • Year and Date: 2016-10-31
  • Invited
• [Presentation] Development of novel nucleoside reverse transcriptase inhibitors active against 4’-ethynyl -2-fluoro-2’-deoxyadenosine (EFdA/MK-8591)-resistant HIV-1s. (2016)
  • Author(s): Maeda K, Takamatsu Y, Kohgo S, Das D, Hattori S, Hayashi H, Stefan G. Sarafianos SG, and Mitsuya H.
  • Organizer: The 15th Awaji International Forum on Infection and Immunity.
  • Place of Presentation: 兵庫
  • Year and Date: 2016-09-06
  • Int'l Joint Research
• [Presentation] 新規の逆転写酵素阻害剤EFdA(MK-8591)に対する薬剤耐性機序の構造学的解析と耐性株に有効な新規薬剤の設計・開発. (2016)
  • Author(s): 前田賢次、高松悠樹、向後悟、服部真一朗、鍬田伸好、林宏典、Debananda Das、満屋裕明.
  • Organizer: 第２６回抗ウイルス療法学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 2016-05-13