Discovery and development of novel anti-HCMV agents targeting the UL89 terminase protein

项目摘要/摘要 人巨细胞病毒(HCMV)感染缺乏完全功能的免疫系统的个人，如 新生儿和移植患者，可能会导致严重和虚弱的后果。美国的大多数人 美国食品和药物管理局批准针对病毒聚合酶和对这些病毒的耐药性的抗巨细胞病毒药物 毒品已经出现了。因此，需要来自新靶点的抗HCMV药物与 或者用现有的聚合酶抑制剂来代替。这项提议的长期目标是开发结构新颖的 针对目前病毒复制以外的病毒复制的直接作用抗病毒药物 批准的药物。为此，一种与病毒基因组密切相关的病毒ATPase/核酸内切酶(PUL89) 包装和病毒组装已被选为抗病毒干预的目标。这样做的目的是 研究目的是鉴定、优化和表征pUL89缓蚀剂支架。为了实现这些目标， 将实现以下具体目标：1.制定和实施筛查试验，以确定pUL89和 人巨细胞病毒抑制剂。这些检测包括pUL89功能的生化检测以及人巨细胞病毒复制 细胞培养中的分析。2.HCMV pUL89抑制剂HIT的产生及优化热门一代将成为中心 围绕内部合成小分子和精选化合物文库的重点筛选。 确认的HITS将通过模拟合成、迭代SAR和体外ADME分析进行优化，以产生 具有良好物理化学性质的强效缓蚀剂。3.先进缓蚀剂的表征 病毒学、结构和药代动力学技术。将分析与pUL89结合的优化的缓蚀剂 并通过结构研究进行提炼。将进行详细的病毒学实验，以确定 作用机制，验证这些抑制剂的靶点，并记录它们与当前FDA的合作能力 批准的药物和抗耐药病毒。此外，优化的体内药动学参数 将确定并使用抑制剂来改善类药物的性能。在概述的研究结束时 在这项建议中，更好地理解了pUL89内切酶活性的作用机制和作用 在病毒复制方面将获得进展，针对pUL89的多种有效临床前候选药物将 发展起来的。

Project Summary/Abstract Human cytomegalovirus (HCMV) infection in individuals lacking a fully functioning immune system, such as newborns and transplant patients, can result in severe and debilitating consequences. The majority of the US Food and Drug Administration approved anti-HCMV drugs target the viral polymerase and resistance to those drugs has appeared. Therefore, anti-HCMV drugs from novel targets are needed for use in combination with, or instead of, current polymerase inhibitors. The long-term goal of this proposal is to develop structurally novel direct-acting antivirals against HCMV targeting aspects of virus replication outside of those for currently approved drugs. Towards this end, a viral ATPase/endonuclease (pUL89) critically involved in viral genome packaging and virus assembly has been chosen as a target for antiviral intervention. The objective of this investigation is to identify, optimize and characterize pUL89 inhibitor scaffolds. To accomplish these goals, the following specific aims will be pursued: 1. Develop and implement screening assays to identify pUL89 and HCMV inhibitors. These assays include biochemical assays for pUL89 function as well as HCMV replication assays in cell culture. 2. HCMV pUL89 inhibitor hit generation and optimization. Hit generation will center around the focused screening of in-house synthetic small molecules and select compound libraries. Confirmed hits will be optimized via analogue synthesis, iterative SAR and in vitro ADME assays to generate potent inhibitors with favorable physicochemical properties. 3. Characterization of advanced inhibitors using virological, structural and pharmacokinetic techniques. Optimized inhibitor binding to pUL89 will be analyzed and refined using structural studies. Detailed virological experiments will be performed to characterize the mechanism of action, verify the target of these inhibitors and document their ability to work with current FDA approved drugs and against resistant virus. In addition, in vivo pharmacokinetic parameters of the optimized inhibitors will be determined and used to improve drug-like properties. At the conclusion of the studies outlined in this proposal, a better understanding of the mechanism of action and role of pUL89 endonuclease activity in virus replication will be gained and multiple potent preclinical drug candidates targeting pUL89 will be developed.