Understanding Envelope Function in HIV-1 Infection: The Design, Synthesis and Validation of Small Molecule HIV-1 Env Inhibitors

了解环境病毒在HIV-1感染中的作用： 小分子HIV-1包膜抑制剂的设计、合成与验证 项目3：总结： 本计划项目(P0-1-GM-06550)的中心目标包括确定和理解 HIV-1膜(Env)三聚体的功能状态：(A)在独立的病毒粒子中；(B)在HIV-1细胞中 进入过程；以及(C)在HIV-1感染的抗体依赖的细胞毒性(ADCC)过程中 细胞。具体地说，我们建议设计、合成和验证与HIV-1环境结合的小分子 糖蛋白(gp120和gp41)，进而调节HIV-1Env的功能状态。这样的目标将 最高要求积累有关环境三聚体的性质和功能的知识 原子分辨率的可能水平。由此产生的理解反过来将允许设计和合成 有效的HIV-1病毒粒子灭活剂、细胞进入抑制剂和HIV-1感染细胞的ADCC增敏剂，它们保持 预防和根除艾滋病毒/艾滋病的承诺。特定的靶向化合物， 预期结合CD4结合口袋，与gp120亚单位的其他区域和/或gp41亚单位结合， 将通过密切合作使用smFRET研究(计划1)、交联质谱、 抑制物结合生物分析数据(项目2、4和核心B)、高分辨率X射线/低温电子显微镜结构(项目5 和合作者Pamela Björkman)和计算建模(核心A)。重要的是，最近的 结合生物学的低温EM和smFRET三聚体可视化技术进展 测量(项目1、2、4和核心B)将有助于阐明小分子 环境调节因子对HIV-1环境蛋白的功能具有调节作用。因此，P01的努力显然将有助于 设计和验证应对艾滋病大流行的新的药理策略。 项目3是本计划项目的综合推力，将专门侧重于计划项目 通过开发和验证与HIV-1环境独特相互作用的新型小分子来实现目标 Trimer(gp120和gp41)，从而提供对环境构象(即环境状态)的洞察 以及它们的生物学意义。双方之间已经并将继续进行的密切合作 五个计划项目和两个核心将明显增强对更多 HIV-1感染细胞的高功能、广谱小分子抑制剂/ADDC增敏剂，以及 引导标记探针以识别、稳定和/或调节导致的重要环境构象变化 进入细胞和/或根除受感染细胞。因此，通过高吞吐量筛选、计算建模、 设计和合成，以及结构分析(smFRET/晶体/低温EM)和生物测定/结合验证 测量，项目3将在该计划项目中发挥核心的、高度集中的作用。

Understanding Env Function in HIV-1 Infection: The Design, Synthesis and Validation of Small Molecule HIV-1 Env Inhibitors Project 3: Summary: The Central Goal of this Program Project (P0-1-GM-06550) comprises the identification and understanding of the functional states of the HIV-1 envelope (Env) trimer: (a) in the standalone virion; (b) during the HIV-1 cell entry process; and (c) during the process of antibody dependent cellular cytotoxicity (ADCC) of HIV-1 infected cells. Specifically we propose to design, synthesize and validate small molecules that bind to the HIV-1 Env glycoproteins (gp120 and gp41), and in turn modulate the HIV-1 Env functional states. Such an objective will require the accumulation of knowledge regarding the nature and function of the Env trimer at the highest possible level of atomic resolution. The derived understanding in turn will permit the design and synthesis of effective HIV-1 virion inactivators, cell entry inhibitors and ADCC sensitizers of HIV-1 infected cells, which hold the promise for both the prevention and eradication of HIV/AIDS. The specific targeted compounds, anticipated to bind the CD4 binding pocket, to other regions of the gp120 subunit and/or to the gp41 subunit, will arise through the close collaborative use of smFRET studies (Program 1), crosslinking-mass spectrometry, inhibitor binding bioassay data (Projects 2, 4 and Core B), high-resolution X-ray/cryoEM structures (Project 5 and collaborator Pamela Björkman) and computational modeling (Core A). Importantly, the recent technological advances in cryoEM and smFRET trimer visualization, in conjunction with biological measurements (Projects 1, 2, 4 and Core B), will permit elucidation of the specific effects that small molecule Env modulators have on the function of the HIV-1 Env. As such, the efforts of the P01 will clearly aid in the design and validation of new pharmacological tactics to address the AIDS pandemic. Project 3, The Synthetic Thrust of this Program Project, will focus specifically on the Program Project goals by developing and validating new classes of small molecules that interact uniquely with the HIV-1 Env trimer (gp120 and gp41), thereby providing insights into the landscape of Env conformations (i.e., Env states) and their biological implications. The close collaboration that has and will continue to occur between each of the five Program Projects and two Cores will clearly enhance the discovery, design, and validation of more highly functional, broad spectrum small molecule inhibitors/ADDC sensitizers of HIV-1 infected cells, as well as lead to labeled probes to identify, stabilize and/or modulate the important Env conformational changes leading to cell entry and/or infected cell eradication. Thus, through high throughput screens, computational modeling, design, and synthesis, along with structural analysis (smFRET/crystal/cryoEM) and bioassay/binding validation measurements, Project 3 will play a central, highly focused role in this Program Project.