Altering Hepatitis B Virus assembly through pharmacological intervention

项目摘要/摘要 病毒是世界范围内发病率和死亡率的主要原因之一，其中乙肝病毒(乙肝病毒)是 其中最具传染性和流行的一种。全球约有2.5亿人患有慢性阻塞性肺病 乙肝病毒感染者，包括在美国的1-200万人。在感染者中，大约有15%-25%的人会患上硒-- 严重的肝脏问题，包括肝硬变、癌症和最终衰竭，每年导致60万人死亡。而当 存在管理慢性乙肝病毒感染的治疗方法，所有针对病毒基因组过程，如逆转录 和DNA复制，由于病毒的持久性和耐药性，没有一种能提供治愈方法。一种很有前途的正交表 消除感染的方法是瞄准乙肝病毒的衣壳，这是包裹病毒基因组的蛋白质外壳。 芬兰实验室之前已经开发出了诱导非功能性分子错误定向组装的化合物 乙肝病毒衣壳，表明小分子有能力影响这些大的多蛋白的结构 装配。虽然对这些化合物的分子间相互作用的结构有很多了解 Identifi事后，关于这种相互作用的动力学几乎没有被探索，并且预测低于... 功能性(抗病毒)结合的标准还有待开发。这两个Defi问题将在#年开始得到解决 这个节目。使用分子动力学(MD)模拟结合对HepAD38细胞的体外实验， PI已经根据以前未知的与乙肝病毒相互作用的基序鉴定了七种化合物，这些基序针对的是fi 衣壳组装工艺。这些化合物结合在两个衣壳蛋白(CP)交界处的已知口袋中。 二聚体，具有中等的抗病毒活性，毒性低。在fi的第一个目标中，使用对接，自由能微扰- PIS将优化先导化合物，以提高其抗乙肝病毒的效果fi。在 第二个目标，使用分子动力学模拟、先进的自由能计算和实验分析 在不同的乙肝病毒蛋白低聚物上，直到并包括整个病毒衣壳，PI将决定有多小 分子干扰衣壳的组装。这两个目标都将使fit受益于使用基于机器学习的Classifi- 阳离子方案以及新的增强型采样方法。在这里学到的关于多小的教训 分子可以将组装途径重定向为非生产性构象，可以推广应用 对其他病毒的影响。

Project Summary/Abstract Viruses are one of the leading causes of morbidity and mortality worldwide, with Hepatitis B Virus (HBV) being one of the most infectious and prevalent among them. Roughly 250 million people worldwide suffer from chronic HBV infection, including 1-2 million in the United States. Of those infected, approximately 15-25% will develop se- rious liver problems, including cirrhosis, cancer, and ultimately failure, leading to 600,000 deaths annually. While therapies exist to manage chronic HBV infection, all target viral genomic processes such as reverse transcription and DNA replication, and none provide a cure due to viral persistence and resistance. A promising orthogonal approach to eliminating infection is to target HBV's capsid, the protein shell that encapsulates the viral genome. The Finn laboratory has previously developed compounds that induce the misdirected assembly of non-functional HBV capsids, showing that small molecules have the power to affect the structures of these large multi-protein assemblies. While much is known about the structures of the intermolecular interactions with these compounds identified after the fact, little about the dynamics of such interactions has been explored, and a predictive under- standing of functional (antiviral) binding has yet to be developed. Both deficiencies will begin to be addressed in this program. Using molecular dynamics (MD) simulations coupled with in vitro experiments on HepAD38 cells, the PIs have identified seven compounds based on motifs previously unknown to interact with HBV that target the capsid assembly process. These compounds bind in a known pocket at the interface of two capsid-protein (Cp) dimers, possess moderate antiviral activity, and low toxicity. In the first aim, using docking, free-energy perturba- tion, and synthetic chemistry, the PIs will optimize the lead compound to improve its efficacy against HBV. In the second aim, using molecular dynamics simulations, advanced free-energy calculations, and experimental assays on different HBV protein oligomers, up to and including the whole virus capsid, the PIs will determine how small molecules interfere with capsid assembly. Both aims will benefit from using a machine-learning-based classifi- cation scheme as well as novel enhanced sampling methods. The lessons learned here concerning how small molecules can re-direct the assembly pathway into unproductive conformations can be generalized for application to other viruses.