Tight-Binding Hydroxypyrazole HIV-1 Nef Inhibitors Suppress Viral Replication in Donor Mononuclear Cells and Reverse Nef-Mediated MHC-I Downregulation
| Autor: | Marianne Carlsen, Thomas E. Smithgall, Colin M. Tice, Wing Fai Li, Jay E. Wrobel, Haibin E. Shi, Allen B. Reitz, Li Chen, Lori A. Emert-Sedlak |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
viruses T cell 030106 microbiology Down-Regulation Virus Replication Major histocompatibility complex Article Cell Line Inhibitory Concentration 50 03 medical and health sciences Drug Development Downregulation and upregulation MHC class I medicine Humans Cytotoxic T cell nef Gene Products Human Immunodeficiency Virus biology Chemistry Histocompatibility Antigens Class I virus diseases Acquired immune system Tissue Donors Cell biology 030104 developmental biology Infectious Diseases medicine.anatomical_structure Anti-Retroviral Agents Viral replication HIV-1 Leukocytes Mononuclear biology.protein Pyrazoles Tyrosine kinase |
| Zdroj: | ACS Infect Dis |
| ISSN: | 2373-8227 |
| DOI: | 10.1021/acsinfecdis.9b00382 |
| Popis: | The HIV-1 Nef accessory factor is critical to the viral life cycle in vivo and promotes immune escape of infected cells via downregulation of cell-surface MHC-I. Previously, we discovered small molecules that bind directly to Nef and block many of its functions, including enhancement of viral infectivity and replication in T cell lines. These compounds also restore cell-surface MHC-I expression in HIV-infected CD4 T cells from AIDS patients, enabling recognition and killing by autologous cytotoxic T lymphocytes (CTLs). In this study, we describe the synthesis and evaluation of a diverse set of analogs based on the original hydroxypyrazole Nef inhibitor core. All analogs were screened for the interaction with recombinant HIV-1 Nef by surface plasmon resonance (SPR) and for antiretroviral activity in TZM-bl reporter cells infected with HIV-1. Active analogs were ranked on the basis of an activity score that integrates three aspects of the SPR data (affinity, residence time, and extent of binding) with antiretroviral activity. The top scoring compounds bound tightly to Nef by SPR, with KD values in the low nM to pM range, and displayed very slow dissociation from their Nef target. These analogs also suppressed HIV-1 replication in donor peripheral blood mononuclear cells (PBMCs) with IC50 values in the 1-10 nM range without cytotoxicity, inhibited Nef-mediated IL-2-inducible tyrosine kinase (Itk) and hematopoietic cell kinase (Hck) activation, and rescued MHC-I downregulation in a Nef-transfected T cell line. The development of Nef inhibitors based on the structure-activity relationships defined here has promise as a new approach to antiretroviral therapy that includes a path to eradication of HIV-infected cells via the adaptive immune response. |
| Databáze: | OpenAIRE |
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