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J Biol Chem. 2015 Apr 17;290(16):10504-17. doi: 10.1074/jbc.M114.626903. Epub 2015 Feb 27.

Identification of a novel HIV-1 inhibitor targeting Vif-dependent degradation of human APOBEC3G protein.

Author information

1
From the Departments of Cancer Immunology and AIDS and Departments of Pathology and.
2
Department of Biology, College of the Holy Cross, Worcester, Massachusetts 01610.
3
Southern Research Institute High Throughput Screening Center, Birmingham, Alabama 35205, and.
4
From the Departments of Cancer Immunology and AIDS and.
5
Cancer Biology, Dana Farber Cancer Institute and.
6
Southern Research Institute, Department of Infectious Disease Research, Frederick, Maryland 21701.
7
From the Departments of Cancer Immunology and AIDS and Neurology (Microbiology), Harvard Medical School, Boston, Massachusetts 02115, dana_gabuzda@dfci.harvard.edu.

Abstract

APOBEC3G (A3G) is a cellular cytidine deaminase that restricts HIV-1 replication by inducing G-to-A hypermutation in viral DNA and by deamination-independent mechanisms. HIV-1 Vif binds to A3G, resulting in its degradation via the 26 S proteasome. Therefore, this interaction represents a potential therapeutic target. To identify compounds that inhibit interaction between A3G and HIV-1 Vif in a high throughput format, we developed a homogeneous time-resolved fluorescence resonance energy transfer assay. A 307,520 compound library from the NIH Molecular Libraries Small Molecule Repository was screened. Secondary screens to evaluate dose-response performance and off-target effects, cell-based assays to identify compounds that attenuate Vif-dependent degradation of A3G, and assays testing antiviral activity in peripheral blood mononuclear cells and T cells were employed. One compound, N.41, showed potent antiviral activity in A3G(+) but not in A3G(-) T cells and had an IC50 as low as 8.4 μM and a TC50 of >100 μM when tested against HIV-1Ba-L replication in peripheral blood mononuclear cells. N.41 inhibited the Vif-A3G interaction and increased cellular A3G levels and incorporation of A3G into virions, thereby attenuating virus infectivity in a Vif-dependent manner. N.41 activity was also species- and Vif-dependent. Preliminary structure-activity relationship studies suggest that a hydroxyl moiety located at a phenylamino group is critical for N.41 anti-HIV activity and identified N.41 analogs with better potency (IC50 as low as 4.2 μM). These findings identify a new lead compound that attenuates HIV replication by liberating A3G from Vif regulation and increasing its innate antiviral activity.

KEYWORDS:

Antiviral Agent; Cytidine Deaminase; High Throughput Screening (HTS); Human Immunodeficiency Virus (HIV); Viral Protein; Virology

PMID:
25724652
PMCID:
PMC4400358
DOI:
10.1074/jbc.M114.626903
[Indexed for MEDLINE]
Free PMC Article

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