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: 10375483
• 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/10375483
• 关键词: Acidity; Acquired Immunodeficiency Syndrome; Active Sites; Address; Affinity; Affinity Chromatography; Bacterial Infections; Binding; Biochemical; Biological Assay; Biological Process; Biotechnology; Carboxylic Acids; Case Study; Cells; Characteristics; Chemicals; Clinical; Collaborations; Data; Development; Disclosure; Disease; Enzymes; Equipment and supply inventories; Funding; HIV; 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; Property; Proteins; Publications; Research; Role; Therapeutic; Therapeutic Uses; Translations; Viral; Virus; Virus Diseases; base; 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; pharmacophore; programs; scaffold; 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.

项目概要/摘要 该研究计划的重点是开发发现金属酶的新策略 抑制剂。金属酶对于许多生物过程至关重要，并且与治疗相关 疾病，包括癌症、细菌/病毒感染、高血压等。尽管普遍存在 金属酶（所有酶的 40% 以上是金属酶）及其在疾病增殖中的关键作用， 新型金属酶抑制剂的开发尚未得到充分探索。 PI（科恩）有 开发了一项将生物无机与药物化学原理相结合的研究计划 被广泛认为是为数不多的专注于金属酶抑制挑战的努力之一。 抑制金属酶的小分子利用金属结合药效团 (MBP) 功能 基团与靶标中的活性位点金属离子结合。在最后一个项目期间，一个集中的 MBP 片段 组装了用于针对金属酶的基于片段的药物发现（FBDD）的库。在这个 更新应用时，这些MBP片段的类药特性将通过应用得到改善 等排体置换。预计这将产生用于识别金属酶的新化学物质 抑制剂，同时获得更广泛的理化特性（例如酸性、亲脂性） 脚手架。这些金属结合电子等排体 (MBI) 将用于改进一类高活性 在上一个项目期间开发的针对流感 N 末端核酸内切酶结构域的抑制剂 聚合酶酸性蛋白（PAN）。虽然对 PAN 核酸内切酶有活性，但摄取特性较差 这些抑制剂导致细胞中对抗病毒的活性不佳。 MBI将用于改善 理化特性，同时保留酶活性，以产生高活性抑制剂 对抗活细胞中的病毒。最后，检查金属酶的靶向活性和选择性 抑制剂、我们的 MBP、MBI 和 PAN 抑制剂将通过细胞热位移测定 (CETSA) 进行检查 和亲和层析。这些实验将验证目标参与度并评估选择性 随着 MBP 开发成全长 PAN 核酸内切酶抑制剂，其性能得到改善。详细的细胞目标 使用这些方法进行金属酶抑制剂的参与数据很少；因此，这些研究将 对于阐明这些治疗化合物的选择性以及临床前景具有重要意义。 之前的项目期间产生了许多合作、专利披露、会议 会议记录和约 13 种出版物。此外，生物技术劳动力的熟练培训生 指导，并将我们的成果转化为初创公司。我们将继续培育 合作发现最好的、一流的金属酶抑制剂，这些抑制剂有可能改善 人类健康。总体而言，该研究计划将继续在识别小型 针对具有挑战性的目标类别的分子抑制剂，对于改善人类健康具有巨大的价值。