Elucidation of TPV's protease dimerization inhibition (PDI) mechanism and development of potent compounds possessing PDI activity

• Project/Area Number: 24791031
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Infectious disease medicine
• Research Institution: Kumamoto Health Science University
• Principal Investigator: AOKI Manabu 熊本保健科学大学, 保健科学部, 講師 (70389542)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: HIV / プロテアーゼ阻害剤 / 薬剤耐性 / HIV-1

Outline of Final Research Achievements

Tipranavir(TPV), a protease inhibitor, has protease dimerization inhibition(PDI)activity. 31 TPV-related resistance mutations were examined to determine whether those mutations affect TPV’s PDI activity. 17 of those mutations reduced TPV’s PDI activity and located in three PR regions critical for PR monomer folding. Furthermore, tests were conducted to determine if TPV binds to monomer PR subunits by using ESI-MS. The tests suggest that TPV binds to both monomer and dimer PR, indicating that TPV likely inhibits PR dimerization through binding to monomer PR subunits. 14 tripeptides carrying identical amino acid sequences as the aforementioned three regions affecting TPV’s PDI activity were synthesized and examined for their influence on PDI activity. Some of the tripeptides were shown to inhibit PR dimerization. Further studies on the mechanisms of dimerization inhibition by compounds should provide better insights required for developing more potent protease dimerization inhibitors.

Research Products

• [Journal Article] Dimerization of HIV-1 protease occurs through two steps relating to the mechanism of protease dimerization inhibition by darunavir. (2014)
  • Author(s): Hayashi H, Takamune N, Nirasawa T, Aoki M, Morishita Y, Das D, Koh Y, Ghosh AK, Misumi S, Mitsuya H.
  • Journal Title: Proc. Natl. Acad. Sci. U S A. Volume: 111 Issue: 33 Pages: 12234-9
  • DOI: 10.1073/pnas.1400027111
  • Peer Reviewed / Open Access
• [Journal Article] Conserved hydrogen-bonding network of P2 bis-tetrahydrofuran containing HIV-1 protease inhibitors (PI) with protease active site amino acid-backbone aid in their activity against PI-resistant HIV. (2014)
  • Author(s): Yedidi RS, Garimella H, Aoki M, Aoki H, Desai DV, Chang SB, Davis DA, Fyvie WS, Kaufman JD, Smith DW, Das D, Wingfield PT, Maeda K, Ghosh AK and Mitsuya H.
  • Journal Title: Antimicrob Agents Chemother Volume: 58 Issue: 7 Pages: 3679-88
  • DOI: 10.1128/aac.00107-14
  • Peer Reviewed
• [Journal Article] Delayed emergence of HIV-1 variants resistant to 4’-ethynyl-2-fluoro-2’-deoxyadenosine: comparative sequential passage study with lamivudine, tenofovir, emtricitabine and BMS-986001 (2013)
  • Author(s): Kenji Maeda, Darshan V Desai, Manabu Aoki, Hirotomo Nakata, Eiichi N Kodama, Hiroaki Mitsuya
  • Journal Title: Antiviral Therapy Volume: 19 Issue: 2 Pages: 179-189
  • DOI: 10.3851/imp2697
  • Peer Reviewed / Open Access
• [Journal Article] P2' benzene carboxylic acid moiety is associated with decrease in cellular uptake: evaluation of novel nonpeptidic HIV-1 protease inhibitors containing P2 bis-tetrahydrofuran moiety. (2013)
  • Author(s): Yedidi RS, Maeda K, Fyvie WS, Steffey M, Davis DA, Palmer I, Aoki M, Kaufman JD, Stahl SJ, Garimella H, Das D, Wingfield PT, Ghosh AK, Mitsuya H.
  • Journal Title: Antimicrobial Agents and Chemotherapy Volume: 57 Issue: 10 Pages: 4920-4927
  • DOI: 10.1128/aac.00868-13
  • Peer Reviewed
• [Journal Article] GRL-0519, a Novel Oxatricyclic Ligand-Containing Nonpeptidic HIV-1 Protease Inhibitor (PI), Potently Suppresses Replication of a Wide Spectrum of Multi-PI-Resistant HIV-1 Variants In Vitro, Antimicrob (2013)
  • Author(s): Amano M, Tojo Y, Gomez PMS, Campbell JR, Das D, Aoki M, Xu CX, Rao KV, Ghosh AK, and Mitsuya H
  • Journal Title: Agents Chemother Volume: 57 Issue: 5 Pages: 2036-2046
  • DOI: 10.1128/aac.02189-12
  • Peer Reviewed
• [Journal Article] Loss of the Protease Dimerization Inhibition Activity of Tipranavir (TPV) and Its Association with the Acquisition of Resistance to TPV by HIV- 1 (2012)
  • Author(s): Manabu Aoki, Matthew L. Danish, Hiromi Aoki- Ogata, Masayuki Amano, Kazuhiko Ide, Debananda Das, Yasuhiro Koh, and Hiroaki Mitsuya
  • Journal Title: Journal of Virology Volume: 86 Issue: 24 Pages: 13384-13396
  • DOI: 10.1128/jvi.07234-11
  • Peer Reviewed