Dual-action virolytic entry inhibitors against HIV-1

 描述（由申请人提供）：该项目的总体目标是开发一种新的靶向HIV-1包膜的范例，以便特异性地抑制病毒， 宿主细胞相遇。独特的HIV-1特异性包膜刺突蛋白，由包埋在病毒膜中的gp 120和gp 41异二聚体的三聚体复合物组成，是暴露在病毒粒子外部的唯一病毒特异性蛋白，因此是中和病毒的有吸引力的靶点。通过靶向Env蛋白的HIV-1灭活可以预防初始感染，并抑制病毒在已感染个体中的传播。最近，我们报道了一种名为CVN-DAVEI的分子的发现，该分子选择性地诱导完全感染性的BaL HIV-1和假型病毒的破坏性裂解。我们的设想是，CVN-DAVEI利用Env复合物固有的亚稳定性来破坏病毒，并且这种亚稳定性是一种致命的弱点，可以针对这种弱点设计一类新的HIV灭活“猎人杀手”分子。虽然HIV-1失活表型很诱人，但我们对DAVEI功能的结构机制的理解还不完整。在这项提案中，我们将确定这一机制，并使用由此产生的机制基础，朝着更小，更临床相关的HIV-1灭活剂先导化合物。我们的项目团队将体现所需的蛋白质科学，分子设计，化学合成和计算结构分析的专业知识，以实现这些总体目标。提出了三个相互关联的具体目标。目标1。确定DAVEI MPER结构域在HIV-1裂解失活中的机制。目标2.通过DAVEI凝集素结构域和DAVEI的化学计量确定Env聚糖结合的机制：Env尖峰相遇。目标3。通过设计新型DAVEI融合构建体，定义Env结构和构象可塑性在DAVEI裂解灭活HIV-1中的作用。总体而言，这项工作将确定双重结合分子对病毒裂解功能的最低要求。反过来，这些结果将扩大我们对病毒亚稳定性的理解，这是HIV-1细胞感染的必要条件，可以用于DAVEI诱导的HIV-1裂解。该项目将产生“猎人杀手”先导化合物，为预防传播和对抗感染开辟新的分子设计策略。最后，工程化到这些抗HIV构建体中的病毒裂解活性将为设计对抗其他亚稳态包膜病毒（如流感、埃博拉和登革热）的分子提供先例。

 DESCRIPTION (provided by applicant): The overall goal of this project is to develop a new paradigm for targeting the HIV-1 envelope in order to inactivate the virus specifically, and before host cell encounter. The uniquely HIV-1-specific envelope spike protein, comprised of the trimeric complex of gp120 and gp41 heterodimers embedded in the virus membrane, is the only virus-specific protein exposed on the outside of the virion particle and thus is an attractive targt for neutralizing the virus. HIV-1 inactivation by targeting the Env protein could prevent initial infection, and suppress virus spread in already-infected individuals. Recently, we reported the discovery of a molecule termed CVN-DAVEI that selectively induced destructive lysis of both fully infectious BaL HIV-1 and pseudotyped virus. Our vision is that CVN-DAVEI takes advantage of the inherent metastability of the Env complex to destroy the virus, and that this metastability is an Achilles' heel against which a new class of HIV- inactivating "hunter-killer" molecules can be designed. While the HIV-1 inactivation phenotype is enticing, our understanding of the structural mechanism of DAVEI function is incomplete. In this proposal, we will determine this mechanism and use the resulting mechanistic underpinning to move towards smaller and more clinically relevant HIV-1 inactivator lead compounds. Our project team will embody the required protein science, molecular design, chemical synthesis and computational structure analysis expertise to accomplish these overarching goals. Three interconnected Specific Aims are proposed. Aim 1. Determine the mechanism of the DAVEI MPER domain in the lytic inactivation of HIV-1. Aim 2. Determine the mechanism of Env glycan engagement by the DAVEI lectin domain and the stoichiometry of DAVEI:Env spike encounter. Aim 3. Define the roles of Env structure and conformational plasticity in the lytic inactivation of HIV-1 by DAVEI via the design of novel DAVEI fusion constructs. Overall, this work will define the minimum requirements for virolytic function by dual-binding molecules. In turn, the results will expand our understanding on how the virus metastability, that is a requirement for HIV-1 cell infection, can be exploited for DAVEI-induced HIV-1 lysis. The project will yield "hunter-killer" lead compounds that will open up new molecular design strategies for preventing transmission and battling infection. Finally, the virolysis activity engineered into these anti-HIV constructs wll provide precedent to design molecules to combat other metastable enveloped viruses, such as influenza, Ebola, and dengue.