Identification of an Antiretroviral Small Molecule That Appears To Be a Host-Targeting Inhibitor of HIV-1 Assembly.
Identification of an Antiretroviral Small Molecule That Appears To Be a Host-Targeting Inhibitor of HIV-1 Assembly.
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DOI：
10.1128/jvi.00883-20
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发表时间：
2021-01-13
影响因子：
5.4
通讯作者：
Lingappa JR
中科院分区：
文献类型：
--
作者：
Reed JC;Solas D;Kitaygorodskyy A;Freeman B;Ressler DTB;Phuong DJ;Swain JV;Matlack K;Hurt CR;Lingappa VR;Lingappa JR
HIV-1 抗逆转录病毒治疗的成功面临着因日益严重的耐药问题而受到损害的风险。因此，需要确定抗逆转录病毒药物，以作用于所用药物未针对的病毒生命周期阶段，例如 HIV-1 Gag 组装事件。为了解决这一差距，我们开发了一种复合筛选，可以概括 HIV-1 组装的细胞内事件，包括促进组装的病毒与宿主的相互作用。这项工作导致鉴定出一种新的化学型，该化学型可能通过作用于组装而在纳摩尔浓度下抑制 HIV-1 复制。该化合物与 Gag 和两种促进衣壳组装的宿主酶共定位。然而，抗性选择并没有导致 gag 中化合物特异性的突变，这表明化学型并不直接针对 Gag。我们假设这种化学型代表了一种一流的病毒产生抑制剂，它通过靶向对 HIV-1 Gag 组装至关重要的病毒宿主复合物发挥作用。鉴于预计多重耐药 HIV-1 的增加，迫切需要开发抗逆转录病毒药物，以作用于目前使用的药物未针对的病毒生命周期阶段。靶向宿主的化合物特别令人感兴趣，因为它们可以提供很高的耐药屏障。在这里，我们报告了抑制 HIV-1 晚期事件的两种相关小分子的鉴定，这是 HIV-1 生命周期的一部分，缺乏有效和特异性的抑制剂。这种化学型是使用无细胞蛋白质合成和组装系统发现的，该系统概括了细胞内宿主催化的病毒衣壳组装途径。这些化合物可抑制人类 T 细胞系和外周血单核细胞中 HIV-1 的复制，并能有效对抗初级分离株。它们可能通过抑制 HIV-1 Gag 组装中的翻译后步骤来减少病毒产生。值得注意的是，该化合物与 HIV-1 Gag 在原位共定位；然而，出乎意料的是，选择实验未能鉴定出 gag 或 pol 中的化合物特异性耐药突变，尽管已知的耐药突变是在平行奈非那韦选择时产生的。因此，我们假设这些化合物不是直接与 Gag 结合，而是定位于组装中间体，即在未成熟的 HIV-1 衣壳组装过程中形成的含有 Gag 和宿主因子的细胞内多蛋白复合物。事实上，受感染细胞的成像显示该化合物与组装中间体 ABCE1 和 DDX6 中发现的两种宿主酶共定位，但其他复合物中没有发现两种宿主蛋白。虽然该化学型的确切靶点和作用机制仍有待确定，但我们的研究结果表明，这些化合物代表了 HIV-1 组装细胞内事件的一流宿主靶向抑制剂。重要性 HIV-1 抗逆转录病毒治疗的成功面临着因日益严重的耐药问题而受到损害的风险。因此，需要确定抗逆转录病毒药物，以作用于所用药物未针对的病毒生命周期阶段，例如 HIV-1 Gag 组装事件。为了解决这一差距，我们开发了一种复合筛选，可以概括 HIV-1 组装的细胞内事件，包括促进组装的病毒与宿主的相互作用。这项工作导致鉴定出一种新的化学型，该化学型可能通过作用于组装而在纳摩尔浓度下抑制 HIV-1 复制。该化合物与 Gag 和两种促进衣壳组装的宿主酶共定位。然而，抗性选择并没有导致 gag 中化合物特异性的突变，这表明化学型并不直接针对 Gag。我们假设这种化学型代表了一种一流的病毒产生抑制剂，它通过靶向对 HIV-1 Gag 组装至关重要的病毒宿主复合物发挥作用。
The success of antiretroviral treatment for HIV-1 is at risk of being undermined by the growing problem of drug resistance. Thus, there is a need to identify antiretrovirals that act on viral life cycle stages not targeted by drugs in use, such as the events of HIV-1 Gag assembly. To address this gap, we developed a compound screen that recapitulates the intracellular events of HIV-1 assembly, including virus-host interactions that promote assembly. This effort led to the identification of a new chemotype that inhibits HIV-1 replication at nanomolar concentrations, likely by acting on assembly. This compound colocalized with Gag and two host enzymes that facilitate capsid assembly. However, resistance selection did not result in compound-specific mutations in gag, suggesting that the chemotype does not directly target Gag. We hypothesize that this chemotype represents a first-in-class inhibitor of virus production that acts by targeting a virus-host complex important for HIV-1 Gag assembly. Given the projected increase in multidrug-resistant HIV-1, there is an urgent need for development of antiretrovirals that act on virus life cycle stages not targeted by drugs currently in use. Host-targeting compounds are of particular interest because they can offer a high barrier to resistance. Here, we report identification of two related small molecules that inhibit HIV-1 late events, a part of the HIV-1 life cycle for which potent and specific inhibitors are lacking. This chemotype was discovered using cell-free protein synthesis and assembly systems that recapitulate intracellular host-catalyzed viral capsid assembly pathways. These compounds inhibit replication of HIV-1 in human T cell lines and peripheral blood mononuclear cells, and are effective against a primary isolate. They reduce virus production, likely by inhibiting a posttranslational step in HIV-1 Gag assembly. Notably, the compound colocalizes with HIV-1 Gag in situ; however, unexpectedly, selection experiments failed to identify compound-specific resistance mutations in gag or pol, even though known resistance mutations developed upon parallel nelfinavir selection. Thus, we hypothesized that instead of binding to Gag directly, these compounds localize to assembly intermediates, the intracellular multiprotein complexes containing Gag and host factors that form during immature HIV-1 capsid assembly. Indeed, imaging of infected cells shows compound colocalized with two host enzymes found in assembly intermediates, ABCE1 and DDX6, but not two host proteins found in other complexes. While the exact target and mechanism of action of this chemotype remain to be determined, our findings suggest that these compounds represent first-in-class, host-targeting inhibitors of intracellular events in HIV-1 assembly. IMPORTANCE The success of antiretroviral treatment for HIV-1 is at risk of being undermined by the growing problem of drug resistance. Thus, there is a need to identify antiretrovirals that act on viral life cycle stages not targeted by drugs in use, such as the events of HIV-1 Gag assembly. To address this gap, we developed a compound screen that recapitulates the intracellular events of HIV-1 assembly, including virus-host interactions that promote assembly. This effort led to the identification of a new chemotype that inhibits HIV-1 replication at nanomolar concentrations, likely by acting on assembly. This compound colocalized with Gag and two host enzymes that facilitate capsid assembly. However, resistance selection did not result in compound-specific mutations in gag, suggesting that the chemotype does not directly target Gag. We hypothesize that this chemotype represents a first-in-class inhibitor of virus production that acts by targeting a virus-host complex important for HIV-1 Gag assembly.