Design of Antivirals and Immunogens Targeting Paramyxoviruses

摘要 病毒感染每年导致数千人死亡和巨大的人道主义负担，然而 与治疗细菌感染的抗生素不同，抗病毒药物很少。目前，针对这一问题的研究方法很少。 产生有效的抗病毒药物。原子分辨率结构信息的不断增加 各种病毒表面蛋白都有可能出现这种情况。此应用程序的总体目标是使用 以副粘病毒表面糖蛋白(PMV)为模型系统生成设计方法学 这将利用广泛的病毒中存在的结构特征，导致强大的 为疫苗接种设计新的疗法、诊断和免疫原的平台。PMV是一种 理想的模型系统作为他们的家庭成员有相同的折叠受体识别，但绑定到非常 不同的宿主细胞受体。 我们最近证明了计算机蛋白质设计可以用来产生新的抗病毒药物。 这在很大程度上中和了各种流感病毒株。这些计算机产生的蛋白质也可以发挥作用 作为高度敏感的诊断。在这些成果的指导下，将实现以下具体目标：(I) 制定针对病毒的一般设计策略：宿主细胞受体相互作用和设计抗病毒药物 Hendra和Nipah病毒作为模型系统；(Ii)通过靶向RSV抑制RSV的膜融合 中间聚合态；以及(Iii)选择性地稳定F-的聚变前后的状态稳定。 并探讨其在传染性和疫苗设计中的作用。 第一个目标是基于观察到被包裹的病毒的许多受体结合部位 在一个凹陷的口袋里，可以逃避免疫系统。计算设计策略 其中专门针对口袋将使开发一种强大的算法来产生抗病毒 结合在这些位点上的蛋白质。第二个目标是基于这样一种假设，即后融合 病毒表面蛋白的结构为靶向其过渡态提供了蓝图。小蛋白质会 被设计成在分子上“堵塞”大多数I型融合蛋白常见的3-螺旋核心结构 因此将提供一种通用方法来抑制I型融合蛋白，包括诸如 如HIV-1、埃博拉、SARS等。最后，目标三的目标是同时对预模型进行建模 以及RSV F蛋白融合后的状态，以产生偏爱一种状态的变体。 将对变异体进行传染性变化检测。捕获的预聚变状态通过不利于 融合后的状态将为免疫原设计的新角度提供依据。如果成功，数据将在 设计将反馈到开发的算法中，导致新的抗病毒药物的快速开发 对抗新出现的流行病。

Abstract Viral infection results in thousands of deaths and an enormous humanitarian burden every year, yet unlike antibiotics for bacterial infection, very few antivirals are available. At present, few methods exist for generating effective antivirals. The increasing availability of atomic resolution structural information of various viral surface proteins promises to chance this. The overall objective of this application is to use the surface glycoproteins of Paramyxoviruses (PMV) as a model system to generate design methodologies that will take advantage of structural features present in a broad range of viruses, resulting in a robust platform for the design of new therapeutics, diagnostics and immunogens for vaccination. PMVs are an ideal model system as their family members have the same fold for receptor recognition yet bind to very different host cell receptors. We recently demonstrated that computational protein design can be used to generate de novo antivirals that broadly neutralize diverse strains of influenza. These computer-generated proteins can also function as highly sensitive diagnostics. Guided by these results, the following specific aims will be pursued: (i) Develop general design strategies to target virus:host cell receptor interactions and design antivirals using Hendra and Nipah Viruses as model systems; (ii) inhibit membrane fusion of RSV by targeting the intermediate fusion states; and (iii) selectively stabilize the pre- and post-fusion state stabilization of the F- protein of RSV and probe their contributions to infectivity and vaccine design. The first aim is based on the observation that many receptor-binding sites of enveloped viruses lay within a recessed pocket, enabling evasion from the immune system. Computational design strategies which specifically target pockets will enable the development a robust algorithm to generate antiviral proteins which bind at these sites. The second aim is based on the hypothesis that the post-fusion structure of viral surface proteins provides the blueprint to targeting their transition state. Small proteins will be designed to molecularly “jam” the 3-helical core structure that is common to most type I fusion proteins and therefore will be provide a general method to inhibit type I fusion proteins, which include viruses such as HIV-1, Ebola, SARS and others. Lastly, the objective of aim three is to simultaneously model the pre- and post-fusion states of the F-protein of RSV to generate variants to favor one state over the other. Variants will be assayed for changes in infectivity. The trapped pre-fusion state stabilized by disfavoring the post-fusion state will provide the basis for a new angle on immunogen design. If successful, data on designs will be fed back into the developed algorithm, leading to rapid development of new antivirals against emerging epidemics.