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

了解HIV-1感染中的Env功能： 小分子HIV-1 Env抑制剂的设计、合成与验证 项目3：摘要： 本计划项目（P0-1-GM-06550）的中心目标包括识别和理解 HIV-1包膜（Env）三聚体的功能状态：（a）在独立的病毒体中;（B）在HIV-1细胞中 进入过程;和（c）在HIV-1感染的抗体依赖性细胞毒性（ADCC）过程中， 细胞具体来说，我们建议设计、合成和验证与HIV-1 Env结合的小分子， 糖蛋白（gp 120和gp 41），并反过来调节HIV-1 Env功能状态。这一目标将 最高需要积累有关Env三聚体性质和功能的知识 可能的原子分辨率。由此得出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敏化剂，其保持 预防和根除艾滋病毒/艾滋病的承诺。特定的目标化合物， 预期结合CD 4结合口袋、gp 120亚基的其它区域和/或gp 41亚基， 将通过密切合作使用smFRET研究（程序1），交联质谱， 抑制剂结合生物测定数据（项目2、4和核心B），高分辨率X射线/冷冻EM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 cryoEM和smFRET三聚体可视化的技术进展，结合生物 测量（项目1、2、4和核心B），将允许阐明小分子 Env调节剂具有对HIV-1 Env的功能。因此，P01的努力显然将有助于 设计和验证新的药物策略，以应对艾滋病的流行。 项目3，本计划项目的综合推力，将特别关注计划项目 通过开发和验证与HIV-1 Env独特相互作用的新型小分子， 三聚体（gp 120和gp 41），从而提供了对Env构象景观的了解（即，环境状态） 及其生物学意义。我们之间的密切合作已经并将继续发生， 五个计划项目和两个核心项目将明显地增强更多的发现、设计和验证 HIV-1感染细胞的高功能、广谱小分子抑制剂/ADDC敏化剂，以及 导致标记的探针以鉴定、稳定和/或调节重要的Env构象变化， 细胞进入和/或感染的细胞根除。因此，通过高通量筛选，计算建模， 设计和合成，沿着结构分析（smFRET/晶体/cryoEM）和生物测定/结合验证 项目3将在本计划项目中发挥核心、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.