Mechanism of resistance to translocation defective reverse transcriptase inhibitors

2013 Fiscal Year Final Research Report

• Project/Area Number: 23590636
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied pharmacology
• Research Institution: Tohoku University
• Principal Investigator: ASHINO Yugo 東北大学, 医学(系)研究科(研究院), 准教授 (20361082)
• Co-Investigator(Kenkyū-buntansha): KODAMA Eiichi 東北大学, 大学院医学系研究科, 講師 (50271151)
• Project Period (FY): 2011-04-28 – 2014-03-31
• Keywords: ウイルス / ＨＩＶ / 薬剤耐性 / 逆転写酵素 / 多剤耐性

Research Abstract

A novel nucleoside reverse transcriptase (RT) inhibitor, 4'-ethynyl-2-fluoro-deoxyadenosine (EFdA) inhibits translocation activity of RT. To elucidate mechanism of the action in detail, resistant HIV-1 and -2 variants to EFdA were induced in vitro. M184I/V in the RT was identified as the primary mutation, while A158T and T165M/R were considered as the secondary mutations. M184I/V much reduced replication kinetics of HIV, but some of the secondary mutations improved the reduced replication. Data obtained from enzymatic assays with recombinant RT well supported those from viral assays. In this study, we provide crucial information of key amino aicds for tranlocation activity and of drug design for novel inhibitors.

Research Products

• [Journal Article] Effect of translocation defective reverse transcriptase inhibitors on the activity of N348I, a connection subdomain drug resistant HIV-1 re- verse transcriptase mutant (2012)
  • Author(s): Michailidis E, Singh K, Ryan EM, Hachiya A, Ong YT, Kirby KA, Marchand B, Kodama EN, Mitsuya H, Parniak MA, Sarafianos SG
  • Journal Title: Cell Mol Biol(Noisy-le-grand) Volume: 58 Pages: 187-95
  • DOI: 10.1155/2012/586401
  • Peer Reviewed
• [Journal Article] Elevated anti-tuberculous glycolipid antibody titres in healthy adults and tuberculosis patients in Thailand (2012)
  • Author(s): Siddiqi UR, Punpunich W, Chuchottaworn C, Jindakul S, Ashino Y, Saitoh H, Okada M, Chotpittayasunondh T, Hattori T
  • Journal Title: Int J Tuberc Lung Dis Volume: 16(4) Pages: 532-538
  • DOI: 10.5588/ijtld.10.0764
  • Peer Reviewed
• [Journal Article] HIV-1 re- verse transcriptase (RT) polymorphism 172K suppresses the effect of clinically relevant drug resistance mutations to both nucleoside and non-nucleoside RT inhibitors (2012)
  • Author(s): Hachiya A, Marchand B, Kirby KA, Michai- lidis E, Tu X, Palczewski K, Ong YT, Li Z, Grif- fin DT, Schuckmann MM, Tanuma J, Oka S, Singh K, Kodama EN, Sarafianos SG
  • Journal Title: J Biol Chem Volume: 287(35) Pages: 29988-99
  • DOI: 10.1074/jbc.M112.351551
  • Peer Reviewed
• [Journal Article] Biochemical, inhibition and inhibitor re- sistance studies of xenotropic murine leukemia virus-related virus reverse transcriptase (2012)
  • Author(s): Ndongwe TP, Adedeji AO, Michailidis E, Ong YT, Hachiya A, Marchand B, Ryan EM, Rai DK, Kirby KA, Whatley AS, Burke DH, Johnson M, Ding S, Zheng YM, Liu SL, Kodama E, Delviks-Frankenberry KA, Pathak VK, Mitsuya H, Parniak MA, Singh K, Sarafianos SG
  • Journal Title: Nucleic Acids Res Volume: 40(1) Pages: 345-59
  • DOI: 10.1093/nar/gkr694
  • Peer Reviewed
• [Journal Article] K70Q adds high-level tenofovir resistance to "Q151M complex" HIV reverse transcriptase through the enhanced discrimination mechanism (2011)
  • Author(s): Hachiya A, Kodama EN, Schuckmann MM, Kirby KA, Michailidis E, Sakagami Y, Oka S, Singh K, Sarafianos SG
  • Journal Title: PLoS One Volume: 6(1) Pages: e16242
  • DOI: 10.1371
  • Peer Reviewed
• [Journal Article] The sugar ring conformation of 4'-ethynyl-2-fluoro-2'-deoxyadenosine and its recognition by the polymerase active site of HIV reverse transcriptase (2011)
  • Author(s): Kirby KA, Singh K, Michailidis E, Marchand B, Kodama EN, Ashida N, Mitsuya H, Parniak MA, Sarafianos SG
  • Journal Title: Cell Mol Biol (Noisy-le-grand) Volume: 57(1) Pages: 40-6
  • URL: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3119259/
  • Peer Reviewed
• [Journal Article] Potent anti-HIV-1 activity of N-HR-derived peptides including a deep pocket-forming region without antagonistic effects
  • Author(s): Watanabe K, Oishi S, Fujii on T-20.Izumi K, N, Matsuoka M, Sarafianos SG, Kodama EN
  • Journal Title: Antivir Chem
  • DOI: 10.3851/IMP1836
• [Presentation] MINICICLIN投与によるヒト免疫不全ウイルス(HIV)持続感染への効果 (2012)
  • Author(s): 芦野有悟、宇佐美修、齊藤弘樹、服部俊夫、賀来満夫
  • Organizer: 第26回日本エイズ学会学術集会
  • Place of Presentation: 神奈川、日本
  • Year and Date: 20121124-26
• [Presentation] Sustained Activity of 4'-Ethynyl Nucleosides to Variants with M184V Mutation in HIV-1 Reverse Transcriptase (2012)
  • Author(s): Fusako Miyamoto,Kumi Kawaji,Toshio Hattori, Hiroaki Mitsuya, Stefan G Sarafianos is, Eiichi N Kodama
  • Organizer: 25th In- ternational Conference on Antiviral Research
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 20120416-19
• [Presentation] Effect of 4'- and 2-NRTI Substitu- tions on the Inhibition Mechanism of HIV Re- verse Transcriptase and Toxicity (2012)
  • Author(s): Eleftherios Michailidis, Jordan Wilkins, Emily M. Ryan, Atsuko Hachiya, Eiichi N. Kodama, Hiroaki Mitsuya, Michael A. Parniak, Stefan G. Sarafianos
  • Organizer: 25th In- ternational Conference on Antiviral Research
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 20120416-19
• [Presentation] The Combination of 4'-Ethynyl-2-Fluoro-2'-Deoxyadenosine with Rilpivirine Shows Synergistic Anti-HIV-1 Activ- ity In Vitro (2012)
  • Author(s): Atsuko Hachiya, Bruno Marchand, Eleftherios Michailidis, Eiichi N Kodama, Michael A Parni- ak, Hiroaki Mitsuya, Shinichi Oka, Stefan G Sarafianos
  • Organizer: 25th In- ternational Conference on Antiviral Research
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 20120416-19