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.0c00888 · 被引用次数：7 · 研究领域：HIV Research and Treatment、HIV/AIDS drug development and treatment、RNA Interference and Gene Delivery

A strategy has been established for the synthesis of a family of bifunctional HIV-1 inhibitor covalent conjugates with the potential to bind simultaneously to both the gp120 and gp41 subunits of the HIV-1 envelope glycoprotein trimeric complex (Env). One component of the conjugates is derived from BNM-III-170, a small-molecule CD4 mimic that binds to gp120. The second component, comprised of the peptide DKWASLWNW ("Trp3"), was derived from the N-terminus of the HIV-1 gp41 Membrane Proximal External Region (MPER) and found previously to bind to the gp41 subunit of Env. The resulting bifunctional conjugates were shown to inhibit virus cell infection with low micromolar potency and to induce lysis of the HIV-1 virion. Crucially, virolysis was found to be dependent on the covalent linkage of the BNM-III-170 and Trp3 domains, as coadministration of a mixture of the un-cross-linked components proved to be nonlytic. However, a significant magnitude of lytic activity was observed in Env-negative and other control pseudoviruses, suggesting parallel mechanisms of action of the conjugates involving Env interaction and direct membrane disruption. Computational modeling suggested strong membrane-binding activity of BNM-III-170, which may underly the nonspecific virolytic effects of the conjugates. To investigate the scope of the membrane effect, cell-based cytotoxicity and membrane permeability assays were performed employing flow cytometry. Here, we observed a dose-dependent and specific...