Tight-Binding Hydroxypyrazole HIV-1 Nef Inhibitors Suppress Viral Replication in Donor Mononuclear Cells and Reverse Nef-Mediated MHC-I Downregulation

被引:17
作者
Shi, Haibin [1 ]
Tice, Colin M. [2 ]
Emert-Sedlak, Lori [1 ]
Chen, Li [1 ]
Li, Wing Fai [1 ,3 ]
Carlsen, Marianne [2 ]
Wrobel, Jay E. [2 ]
Reitz, Allen B. [2 ]
Smithgall, Thomas E. [1 ]
机构
[1] Univ Pittsburgh, Dept Microbiol & Mol Genet, Sch Med, Suite 523 Bridgeside Point II,450 Technol Dr, Pittsburgh, PA 15219 USA
[2] Fox Chase Chem Divers Ctr Inc, Penn Biotechnol Ctr, 3805 Old Easton Rd, Doylestown, PA 18902 USA
[3] Tsinghua Univ, Sch Med, Beijing 100084, Peoples R China
来源
ACS INFECTIOUS DISEASES | 2020年 / 6卷 / 02期
基金
美国国家卫生研究院;
关键词
HIV/AIDS; antiretroviral drug development; HIV-1 Nef inhibitors; HIV accessory proteins; surface plasmon resonance; CRYSTAL-STRUCTURE; SRC; INFECTION; KINASES; ACTIVATION; SERINC5; PROTEIN; ASSAY; HCK; ITK;
D O I
10.1021/acsinfecdis.9b00382
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
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 K-D 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.
引用
收藏
页码:302 / 312
页数:21
相关论文
共 42 条
  • [31] How HIV-1 Nef hijacks the AP-2 clathrin adaptor to downregulate CD4
    Ren, Xuefeng
    Park, Sang Yoon
    Bonifacino, Juan S.
    Hurley, James H.
    [J]. ELIFE, 2014, 3
  • [32] HIV-1 Nef promotes infection by excluding SERINC5 from virion incorporation
    Rosa, Annachiara
    Chande, Ajit
    Ziglio, Serena
    De Sanctis, Veronica
    Bertorelli, Roberto
    Goh, Shih Lin
    McCauley, Sean M.
    Nowosielska, Anetta
    Antonarakis, Stylianos E.
    Luban, Jeremy
    Santoni, Federico Andrea
    Pizzato, Massimo
    [J]. NATURE, 2015, 526 (7572) : 212 - +
  • [33] Saksela K, 1997, FRONT BIOSCI, V2, pd606
  • [35] Targeting the Latent Reservoir for HIV-1
    Sengupta, Srona
    Siliciano, Robert F.
    [J]. IMMUNITY, 2018, 48 (05) : 872 - 895
  • [36] The Accessory Factor Nef Links HIV-1 to Tec/Btk Kinases in an Src Homology 3 Domain-dependent Manner
    Tarafdar, Sreya
    Poe, Jerrod A.
    Smithgall, Thomas E.
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2014, 289 (22) : 15718 - 15728
  • [37] Current status of antiretroviral therapy
    Temesgen, Zelalem
    Warnke, David
    Kasten, Mary J.
    [J]. EXPERT OPINION ON PHARMACOTHERAPY, 2006, 7 (12) : 1541 - 1554
  • [38] HIV-1 Nef selectively activates Src family kinases Hck, Lyn, and c-Src through direct SH3 domain interaction
    Trible, Ronald P.
    Emert-Sedlak, Lori
    Smithgall, Thomas E.
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2006, 281 (37) : 27029 - 27038
  • [39] Discovery of a diaminoquinoxaline benzenesulfonamide antagonist of HIV-1 Nef function using a yeast-based phenotypic screen
    Trible, Ronald P.
    Narute, Purushottam
    Emert-Sedlak, Lori A.
    Alvarado, John Jeff
    Atkins, Katelyn
    Thomas, Laurel
    Kodama, Toshiaki
    Yanamala, Naveena
    Korotchenko, Vasiliy
    Day, Billy W.
    Thomas, Gary
    Smithgall, Thomas E.
    [J]. RETROVIROLOGY, 2013, 10
  • [40] SERINC3 and SERINC5 restrict HIV-1 infectivity and are counteracted by Nef
    Usami, Yoshiko
    Wu, Yuanfei
    Goettlinger, Heinrich G.
    [J]. NATURE, 2015, 526 (7572) : 218 - +