Oxazole-Benzenesulfonamide Derivatives Inhibit HIV-1 Reverse Transcriptase Interaction with Cellular eEF1A and Reduce Viral Replication.

Abstract

HIV-1 replication requires direct interaction between HIV-1 reverse transcriptase (RT) and cellular eukaryotic translation elongation factor 1A (eEF1A). Our previous work showed that disrupting this interaction inhibited HIV-1 uncoating, reverse transcription, and replication, indicating its potential as an anti-HIV-1 target. In this study, we developed a sensitive, live-cell split-luciferase complementation assay (NanoBiT) to quantitatively measure inhibition of HIV-1 RT interaction with eEF1A. We used this to screen a small molecule library and discovered small-molecule oxazole-benzenesulfonamides (C7, C8, and C9), which dose dependently and specifically inhibited the HIV-1 RT interaction with eEF1A. These compounds directly bound to HIV-1 RT in a dose-dependent manner, as assessed by a biolayer interferometry (BLI) assay, but did not bind to eEF1A. These oxazole-benzenesulfonamides did not inhibit enzymatic activity of recombinant HIV-1 RT in a homopolymer assay but did inhibit reverse transcription and infection of both wild-type (WT) and nonnucleoside reverse transcriptase inhibitor (NNRTI)-resistant HIV-1 in a dose-dependent manner in HEK293T cells. Infection of HeLa cells was significantly inhibited by the oxazole-benzenesulfonamides, and the antiviral activity was most potent against replication stages before 8 h postinfection. In human primary activated CD4+ T cells, C7 inhibited HIV-1 infectivity and replication up to 6 days postinfection. The data suggest a novel mechanism of HIV-1 inhibition and further elucidate how the RT-eEF1A interaction is important for HIV-1 replication. These compounds provide potential to develop a new class of anti-HIV-1 drugs to treat WT and NNRTI-resistant strains in people infected with HIV.IMPORTANCE Antiretroviral drugs protect many HIV-positive people, but their success can be compromised by drug-resistant strains. To combat these strains, the development of new classes of HIV-1 inhibitors is essential and a priority in the field. In this study, we identified small molecules that bind directly to HIV-1 reverse transcriptase (RT) and inhibit its interaction with cellular eEF1A, an interaction which we have previously identified as crucial for HIV-1 replication. These compounds inhibit intracellular HIV-1 reverse transcription and replication of WT HIV-1, as well as HIV-1 mutants that are resistant to current RT inhibitors. A novel mechanism of action involving inhibition of the HIV-1 RT-eEF1A interaction is an important finding and a potential new way to combat drug-resistant HIV-1 strains in infected people.

• Authors: Daniel J Rawle, Dongsheng Li, Zhonglan Wu, Lu Wang, Marcus Choong, Mary Lor, Robert C Reid, David P Fairlie, Jonathan Harris, Gilda Tachedjian, Sally-Ann Poulsen, David Harrich
• Journal: Journal of virology (J Virol) Vol. 93 Issue 12 (06 15 2019) ISSN: 1098-5514 [Electronic] United States
• PMID: 30918071 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2019 American Society for Microbiology.

Chemical References

• Anti-HIV Agents
• EEF1A1 protein, human
• Oxazoles
• Peptide Elongation Factor 1
• Reverse Transcriptase Inhibitors
• Sulfonamides
• benzenesulfonamide
• reverse transcriptase, Human immunodeficiency virus 1
• HIV Reverse Transcriptase

Topics

• Anti-HIV Agents (pharmacology)
• HEK293 Cells
• HIV Infections (drug therapy)
• HIV Reverse Transcriptase (drug effects, metabolism)
• HIV-1 (physiology)
• HeLa Cells
• Humans
• Oxazoles (metabolism, pharmacology)
• Peptide Elongation Factor 1 (metabolism)
• Reverse Transcriptase Inhibitors (pharmacology)
• Reverse Transcription (drug effects)
• Sulfonamides (metabolism, pharmacology)
• Virus Replication (drug effects)