Translocation of the HIV-1 pre-integration complex (PIC) into the nucleus of an infected cell is a critical step in viral replication. During the previous funding period of this grant, we demonstrated that arylene bis(methylketone) compounds effectively and selectively inhibit HIV-1 nuclear translocation and replication of the virus in primary cells. We also demonstrated that the molecular mechanism of action of these compounds involves drug interaction with both reverse transcriptase (RT) and matrix (MA) proteins. We initiated studies of viral resistance to these compounds and obtained partially resistant isolate. We now propose to clone and genetically characterize this isolate. identification of mutations responsible for resistance to arylene bis(methylketone) compounds would validate the proposed mechanism of their activity and help evaluate their usefulness for future clinical applications as an addition to multi-drug cocktail. In an attempt to eliminate dependence of nuclear import inhibitors on interaction with RT, we designed a novel class of naphthoquinone compounds. Preliminary results demonstrated that some of these compounds specifically inhibited HIV-1 nuclear import in vitro and blocked viral replication in macrophage cultures without cytotoxic effects. We propose studies to determine the molecular mechanism of action of naphthoquinone compounds. We also expect these new compounds to be active against the virus resistant to arylene bis(methylketone) drugs. Studies by our and other groups demonstrated the important role of Vpr in HIV-I nuclear import and replication in macrophages. This Vpr function depends on its interaction with the cellular nuclear import factor, karyopherin. Based on our preliminary results that identify interacting sites on Vpr and karyopherin molecules, we propose to design and test compounds that will inhibit interaction between these proteins. Upon completion, these studies are expected to define potent anti-HIV compounds working through a novel mechanism different from that of any other currently used drug.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI040386-04A1
Application #
6313481
Study Section
AIDS and Related Research 8 (AARR)
Program Officer
Miller, Roger H
Project Start
1997-04-15
Project End
2001-08-31
Budget Start
2001-02-01
Budget End
2001-08-31
Support Year
4
Fiscal Year
2001
Total Cost
$176,198
Indirect Cost
Name
Picower Institute for Medical Research
Department
Type
DUNS #
City
Manhasset
State
NY
Country
United States
Zip Code
11030
Levin, Aviad; Loyter, Abraham; Bukrinsky, Michael (2011) Strategies to inhibit viral protein nuclear import: HIV-1 as a target. Biochim Biophys Acta 1813:1646-53
Li, Ge; Elder, Robert T; Dubrovsky, Larisa et al. (2010) HIV-1 replication through hHR23A-mediated interaction of Vpr with 26S proteasome. PLoS One 5:e11371
Iordanskiy, Sergey N; Bukrinsky, Michael I (2009) Analysis of viral and cellular proteins in HIV-1 reverse transcription complexes by co-immunoprecipitation. Methods Mol Biol 485:121-34
Iordanskiy, Sergey; Berro, Reem; Altieri, Maria et al. (2006) Intracytoplasmic maturation of the human immunodeficiency virus type 1 reverse transcription complexes determines their capacity to integrate into chromatin. Retrovirology 3:4
Haffar, Omar; Dubrovsky, Larisa; Lowe, Richard et al. (2005) Oxadiazols: a new class of rationally designed anti-human immunodeficiency virus compounds targeting the nuclear localization signal of the viral matrix protein. J Virol 79:13028-36
Al-Abed, Yousef; Dubrovsky, Larisa; Ruzsicska, Bela et al. (2002) Inhibition of HIV-1 nuclear import via schiff base formation with arylene bis(methylketone) compounds. Bioorg Med Chem Lett 12:3117-9
Glushakova, S; Dubrovsky, L; Grivel, J et al. (2000) Small molecule inhibitor of HIV-1 nuclear import suppresses HIV-1 replication in human lymphoid tissue ex vivo: a potential addition to current anti-HIV drug repertoire. Antiviral Res 47:89-95
Bukrinsky, M I; Haffar, O K (1999) HIV-1 nuclear import: in search of a leader. Front Biosci 4:D772-81
Haffar, O K; Smithgall, M D; Popov, S et al. (1998) CNI-H0294, a nuclear importation inhibitor of the human immunodeficiency virus type 1 genome, abrogates virus replication in infected activated peripheral blood mononuclear cells. Antimicrob Agents Chemother 42:1133-8
Bukrinsky, M I; Haffar, O K (1997) HIV-1 nuclear import: in search of a leader. Front Biosci 2:d578-87