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

发现和开发针对 UL89 终止酶蛋白的新型抗 HCMV 药物

• 批准号: 10318943
• 负责人: Robert James Geraghty
• 金额: $ 65.45万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-25 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318943
• 关键词: ATP phosphohydrolase; Acquired Immunodeficiency Syndrome; Active Sites; Antiviral Agents; Binding; Biochemical; Biochemistry; Biological Assay; Brain Injuries; Cell Culture Techniques; Cell Survival; Chemistry; Cidofovir; Clinic; Collaborations; Complex; Crystallization; Cytomegalovirus; DNA Packaging; Data; Development; Disease; Drug Kinetics; Drug Targeting; Drug resistance; Enzymes; FDA approved; Fetal Development; Foscarnet; Foundations; Ganciclovir; Generations; Goals; Human; Immunocompromised Host; In Vitro; Individual; Infection; Infectious Agent; Intervention; Investigation; Libraries; Metals; Molecular Target; Morbidity - disease rate; Mutation; Mutation Analysis; Newborn Infant; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Polymerase; Prodrugs; Production; Property; Prophylactic treatment; Proteins; Publishing; Research; Resistance; Role; Signal Transduction; Structure; Techniques; Toxic effect; Transplant Recipients; Transplantation; United States Food and Drug Administration; Valganciclovir; Viral; Viral Genome; Virus; Virus Assembly; Virus Replication; Work; allograft rejection; analog; base; cost; drug candidate; endonuclease; experimental study; hearing impairment; high throughput screening; immune system function; improved; in vivo; inhibitor; innovation; interdisciplinary approach; mortality; new therapeutic target; novel; novel therapeutics; pharmacophore; post-transplant; pre-clinical; prevent; resistant strain; scaffold; screening; side effect; small molecule; standard of care; structural biology; success; targeted agent; terminase; tool; viral genomics; viral resistance; virology; virtual screening

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.

项目总结/摘要 人巨细胞病毒（HCMV）感染缺乏完全功能的免疫系统的个体，如 新生儿和移植患者，可能导致严重和衰弱的后果。美国多数 美国食品和药物管理局批准的抗HCMV药物靶向病毒聚合酶和耐药性 毒品出现了。因此，需要来自新靶点的抗HCMV药物与以下药物组合使用， 或代替现有的聚合酶抑制剂。该提案的长期目标是开发结构新颖的 针对HCMV的直接作用的抗病毒药物靶向病毒复制的方面， 批准的药物。为此，病毒ATP酶/内切核酸酶（pUL 89）关键地参与病毒基因组， 包装和病毒装配被选为抗病毒干预的目标。的目的 研究是鉴定、优化和表征pUL 89抑制剂支架。为了实现这些目标， 具体目标如下：开发和实施筛选试验，以鉴定pUL 89和 HCMV抑制剂。这些试验包括pUL 89功能以及HCMV复制的生化试验 细胞培养中的测定。2. HCMV pUL 89抑制剂命中生成和优化。点击生成将集中 围绕内部合成小分子和选择化合物库的集中筛选。 确认的命中将通过类似物合成、迭代SAR和体外ADME测定进行优化，以产生 具有良好物理化学性质的有效抑制剂。3.先进抑制剂的表征， 病毒学、结构和药代动力学技术。将分析优化的抑制剂与pUL 89的结合 并利用结构研究进行了改进。将进行详细的病毒学实验以表征 作用机制，验证这些抑制剂的靶点，并记录其与当前FDA合作的能力 批准的药物和抵抗耐药病毒。此外，优化的药物在体内的药代动力学参数 抑制剂将被确定并用于改善药物样性质。在概述的研究结束时， 在这个建议中，更好地理解pUL 89内切酶活性的作用机制和作用， 在病毒复制中将获得，靶向pUL 89的多种有效的临床前候选药物将被 开发