Metal-binding Isosteres for Influenza Endonuclease Inhibitors and Beyond

流感核酸内切酶抑制剂及其他药物的金属结合等排体

• 批准号: 10594905
• 负责人: SETH M COHEN
• 金额: $ 37.07万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594905
• 关键词: Acidity; Active Sites; Address; Affinity; Affinity Chromatography; Bacterial Infections; Binding; Biochemical; Biological Assay; Biological Process; Biotechnology; Carboxylic Acids; Case Study; Cells; Characteristics; Chemicals; Clinical; Collaborations; Cytoprotection; Data; Development; Disclosure; Disease; Enzymes; Equipment and supply inventories; Funding; HIV/AIDS; Health; Human; Hypertension; In Vitro; Influenza; Ions; Laboratories; Legal patent; Length; Libraries; Malignant Neoplasms; Mammalian Cell; Measures; Mentors; Metal Ion Binding; Metals; Methods; Modeling; Multi-Drug Resistance; N-terminal; Nature; Pharmaceutical Chemistry; Pharmaceutical Preparations; Play; Polymerase; Prevalence; Proliferating; Property; Proteins; Publications; Research; Role; Therapeutic; Therapeutic Uses; Translations; Viral; Viral Physiology; Virus; Virus Diseases; cellular targeting; clinically relevant; design; drug development; drug discovery; endonuclease; experimental study; flu; flu activity; functional group; human disease; improved; in vitro activity; influenzavirus; inhibitor; innovation; insight; lipophilicity; metalloenzyme; meter; pharmacophore; programs; scaffold; skills; small molecule; small molecule inhibitor; small molecule therapeutics; symposium; uptake; Acidity; Active Sites; Address; Affinity; Affinity Chromatography; Bacterial Infections; Binding; Biochemical; Biological Assay; Biological Process; Biotechnology; Carboxylic Acids; Case Study; Cells; Characteristics; Chemicals; Clinical; Collaborations; Cytoprotection; Data; Development; Disclosure; Disease; Enzymes; Equipment and supply inventories; Funding; HIV/AIDS; Health; Human; Hypertension; In Vitro; Influenza; Ions; Laboratories; Legal patent; Length; Libraries; Malignant Neoplasms; Mammalian Cell; Measures; Mentors; Metal Ion Binding; Metals; Methods; Modeling; Multi-Drug Resistance; N-terminal; Nature; Pharmaceutical Chemistry; Pharmaceutical Preparations; Play; Polymerase; Prevalence; Proliferating; Property; Proteins; Publications; Research; Role; Therapeutic; Therapeutic Uses; Translations; Viral; Viral Physiology; Virus; Virus Diseases; cellular targeting; clinically relevant; design; drug development; drug discovery; endonuclease; experimental study; flu; flu activity; functional group; human disease; improved; in vitro activity; influenzavirus; inhibitor; innovation; insight; lipophilicity; metalloenzyme; meter; pharmacophore; programs; scaffold; skills; small molecule; small molecule inhibitor; small molecule therapeutics; symposium; uptake

PROJECT SUMMARY/ABSTRACT This research program is focused on developing new strategies for the discovery of metalloenzyme inhibitors. Metalloenzymes are essential to numerous biological processes and are relevant to treating diseases, including cancer, bacterial/viral infections, hypertension, and others. Despite the prevalence of metalloenzymes (>40% of all enzymes are metalloenzymes) and their critical role in disease proliferation, the development of new metalloenzyme inhibitors is extremely underexplored. The PI (Cohen) has developed a research program that combines the principles of bioinorganic with medicinal chemistry and is widely recognized as one of the few efforts focused on the challenges of metalloenzyme inhibition. Small molecules that inhibit metalloenzymes utilize a metal-binding pharmacophore (MBP) functional group to bind to the active site metal ion(s) in the target. In the last project period, a focused MBP fragment library for use in fragment-based drug discovery (FBDD) against metalloenzymes was assembled. In this renewal application, the drug-like features of these MBP fragments will be improved by the application of isostere replacement. This is expected to yield new chemical matter for identifying metalloenzyme inhibitors, while accessing a wider range of physicochemical properties (e.g., acidity, lipophilicity) in these scaffolds. These metal-binding isosteres (MBIs) will then be used to improve a class of highly active inhibitors developed during the last project period against the influenza N-terminal endonuclease domain of the polymerase acidic protein (PAN). Although active against PAN endonuclease, the poor uptake properties of these inhibitors have led to suboptimal activity against the virus in cells. MBIs will be used to improve physicochemical properties, while retaining enzyme-based activity, to produce highly active inhibitors against the virus in live cells. Finally, to examine the on-target activity and selectivity of metalloenzyme inhibitors, our MBPs, MBIs, and PAN inhibitors will be examined by Cellular Thermal Shift Assay (CETSA) and affinity chromatography. These experiments will verify target engagement and evaluate how selectivity improves as the MBP is developed into a full-length PAN endonuclease inhibitor. Detailed cellular target engagement data using these methods for metalloenzyme inhibitors is scarce; therefore, these studies will be valuable for clarifying the selectivity, and hence the clinical prospects, of these therapeutic compounds. The previous project period generated many collaborations, patent disclosures, conference proceedings, and ~13 publications. In addition, skilled trainees for the biotechnology workforce were mentored, and translation of our results into startup companies was achieved. We will continue to nurture collaborations to discover best- and first-in-class metalloenzyme inhibitors that have the potential to improve human health. Overall, this research program will continue to play a leading role in identifying small molecule inhibitors for a challenging target class that has great merit for improving human health.

项目摘要/摘要 该研究计划的重点是制定新策略以发现金属酶 抑制剂。金属酶对于众多生物过程至关重要，与治疗有关 包括癌症，细菌/病毒感染，高血压等疾病。尽管流行 金属酶（>所有酶的40％是金属酶）及其在疾病增殖中的关键作用， 新的金属酶抑制剂的开发极为充实。 PI（Cohen）具有 制定了一个研究计划，该计划将生物有机剂的原理与药物化学结合在一起，并且 广泛认为是少数努力的努力之一，这些努力集中在金属酶抑制的挑战上。 抑制金属酶使用金属结合药效团（MBP）功能的小分子 组与目标中的活性位点金属离子结合。在最后一个项目时期，集中的MBP片段 组装了用于基于碎片的药物发现（FBDD）针对金属酶的库。在这个 续订应用，这些MBP片段的类似药物的特征将通过应用来提高 ISOSTORE替代。预计这将产生新的化学物质来鉴定金属酶 抑制剂，在这些抑制剂中访问这些物理化学特性（例如酸度，亲脂性）的同时 脚手架。然后将使用这些金属结合等值剂（MBI）来改善一类高度活性 在最后一个项目时期对抑制剂的抑制剂针对流感的N末端核酸内切酶结构域 聚合酶酸性蛋白（PAN）。虽然对PAN核酸内切酶有活跃，但摄入效果差 这些抑制剂已导致对细胞中病毒的次优活性。 MBI将用于改进 物理化学特性，同时保留基于酶的活性，以产生高度活性的抑制剂 针对活细胞中的病毒。最后，检查靶标活动和金属酶的选择性 抑制剂，我们的MBP，MBI和PAN抑制剂将通过细胞热移测定法检查（CETSA） 和亲和力色谱。这些实验将验证目标参与并评估选择性 随着MBP发展为全长PAN核酸酶抑制剂，改善。详细的细胞靶 使用这些方法用于金属酶抑制剂的参与数据很少。因此，这些研究将 对于阐明这些治疗性化合物的选择性以及临床前景的有价值。 上一个项目时期产生了许多合作，专利披露，会议 程序，约有13个出版物。此外，生物技术劳动力的熟练学员是 受到指导，并将结果转换为初创公司。我们将继续养育 发现有可能改进的最佳和一流金属酶抑制剂的合作 人类健康。总体而言，该研究计划将继续在确定小型 具有挑战性目标类别的分子抑制剂，具有改善人类健康的功能。