HIV-1 Env-Dependent Cell Killing by Bifunctional Small-Molecule/Peptide Conjugates

作者：Althea Gaffney, Aakansha Nangarlia, Charles G. Ang, Steven T. Gossert, Adel A. Rashad, Md. Alamgir Hossain, Cameron F. Abrams, Amos B. Smith, Irwin Chaiken · 发表于：ACS Chemical Biology · 年份：2021 · DOI：10.1021/acschembio.1c00039 · 研究领域：RNA Interference and Gene Delivery、HIV Research and Treatment、Immunotherapy and Immune Responses

A strategy\nhas been established for the synthesis of a family of\nbifunctional HIV-1 inhibitor covalent conjugates with the potential\nto bind simultaneously to both the gp120 and gp41 subunits of the\nHIV-1 envelope glycoprotein trimeric complex (Env). One component\nof the conjugates is derived from BNM-III-170, a small-molecule CD4\nmimic that binds to gp120. The second component, comprised of the\npeptide DKWASLWNW (“Trp3”), was derived from the N-terminus\nof the HIV-1 gp41 Membrane Proximal External Region (MPER) and found\npreviously to bind to the gp41 subunit of Env. The resulting bifunctional\nconjugates were shown to inhibit virus cell infection with low micromolar\npotency and to induce lysis of the HIV-1 virion. Crucially, virolysis\nwas found to be dependent on the covalent linkage of the BNM-III-170\nand Trp3 domains, as coadministration of a mixture of the un-cross-linked\ncomponents proved to be nonlytic. However, a significant magnitude\nof lytic activity was observed in Env-negative and other control pseudoviruses,\nsuggesting parallel mechanisms of action of the conjugates involving\nEnv interaction and direct membrane disruption. Computational modeling\nsuggested strong membrane-binding activity of BNM-III-170, which may\nunderly the nonspecific virolytic effects of the conjugates. To investigate\nthe scope of the membrane effect, cell-based cytotoxicity and membrane\npermeability assays were performed employing flow cytometry. Here,\nwe observed a dose-...