Assembly and inhibition thermodynamics

主要研究者/项目负责人（最后一名、第一名、中间名）：Chaiken、欧文M / Freire、Ernesto 产品说明： 病毒包膜糖蛋白gp 120负责HIV-1感染的初始事件。的结合 gp 120与细胞表面受体CD 4的结合诱导了一系列变构事件，最终导致gp 120与CD 4的融合。 病毒和细胞膜。整个系列事件定义了不同的干预点， 过程可以中断，因此为抑制剂的形成提供了各种机会。这些 干预点定义了两类抑制剂：竞争性抑制GP 120结合的化合物 其细胞伴侣和化合物阻断变构信号和下游激活。的 针对任何这些干预点的抑制剂的合理设计和优化需要精确的 对GP 120的结构能量学和构象稳定性，其结合相互作用， 变构途径和抑制剂靶向的潜在位点。此外，由于绝大多数HIV-1 感染发生在撒哈拉以南非洲，那里的主要病毒亚型是C和A，而不是B亚型 负责在美国和西欧的感染，重要的是，抑制剂是 对这些亚型有效，因为它们的基因组差异高达30%，包括gp 120。 这些是该项目所处理的主要问题，可归纳为以下具体目标： 1.热力学表征的结构稳定性和结合之间的合作联系 通过利用微量热和基于结构的热力学的组合， 问题研究 2.竞争性抑制剂热力学指南的发展。如何抑制gp 120 结合而不触发变构激活级联反应？ 3.变构抑制剂热力学指南的发展。潜在结合的鉴别 可以靶向阻断GP 120变构途径的位点。 4.别构抑制剂BMS-806及其类似物的表征，其是临床上唯一的gp 120抑制剂 问题研究BMS-806结合位点的鉴定。确定房舍管理系统的关键功能- 806.基于不同化学支架的BMS-806样抑制剂的开发。 5.开发对不同的抗肿瘤药物有效的gp 120抑制剂的热力学指南 HIV-1亚型，对潜在的耐药突变体表现出低易感性。 这些研究涉及实验热力学测量（等温滴定）的组合 量热法和差示扫描量热法）和基于结构的热力学分析。

Principal Investigator/Program Director (Last, First, Middle): Chaiken, Irwin M / Freire, Ernesto DESCRIPTION: The viral envelope glycoprotein gp120 is responsible for the initial events in HIV-1 infection. The binding of gp120 to the cell surface receptor CD4 induces a series of allosteric events that culminate with the fusion of the viral and cell membranes. The entire series of events defines different intervention points at which the process can be interrupted, therefore providing various opportunities for inhibitor development. These intervention points define two classes of inhibitors: compounds that competitively inhibit the binding of gp120 to its cellular partners and compounds that block allosteric signaling and downstream activation. The rational design and optimization of.inhibitors directed at any of these intervention points requires a precise knowledge of the structural energetics and conformational stability of gp120, its binding interactions, allosteric pathways and potential sites for inhibitor targeting. In addition, since the vast majority of HIV-1 infection occurs in Sub-Saharan Africa, where the main viral subtypes are C and A rather than the B subtype responsible for the infection in the United States and Western Europe, it is important that inhibitors are effective against those subtypes since they differ by as much as 30% in their genomes, including gp120. these are the main issues addressed in this project and can be summarized in the following specific aims: 1. Thermodynamic characterization of the structural stability and cooperative linkage between binding sites in gp120 by utilizing a combination of microcalorimetric and structure-based thermodynamic studies. 2. Development of thermodynamic guidelines for competitive inhibitors. How do we inhibit gp120 binding without triggering the allosteric activation cascade? 3. Development of thermodynamic guidelines for allosteric inhibitors. Identification of potential binding sites that can be targeted for blocking gp120 allosteric pathways. 4. Characterization of the allosteric inhibitor BMS-806 and analogs, the only gp120 inhibitor in clinical studies. Identification of the BMS-806 binding site. Identification of the critical functionalities in BMS- 806. Development of BMS-806-like inhibitors based upon different chemical scaffolds. 5. Development of thermodynamic guidelines for gp120 inhibitors that are effective against different HIV-1 subtypes and exhibit low susceptibility to potential drug resistant mutants. The studies involve a combination of experimental thermodynamic measurements (isothermal titration calorimetry and differential scanning calorimetry) and structure-based thermodynamic analysis.