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: 10113523
• 负责人: SETH M COHEN
• 金额: $ 37.04万
• 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/10113523
• 关键词: Acidity; Acquired Immunodeficiency Syndrome; Active Sites; Address; Affinity; Affinity Chromatography; Antiviral Agents; 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/antagonist; 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.

项目摘要/摘要 这项研究计划致力于开发发现金属酶的新策略。 抑制剂。金属酶在许多生物过程中是必不可少的，并与治疗相关。 疾病，包括癌症、细菌/病毒感染、高血压等。尽管流行的是 金属酶(40%的酶是金属酶)及其在疾病扩散中的关键作用， 新的金属酶抑制剂的开发极不充分。少年派(科恩)有 开发了一项将生物无机物原理与药物化学相结合的研究计划，并 被广泛认为是关注金属酶抑制挑战的为数不多的努力之一。 抑制金属酶的小分子利用金属结合药效团(MBP)功能 与靶标中活性中心金属离子(S)结合的基团。在上一个项目期间，一个重点关注的MBP片段 组装了用于抗金属酶片段药物发现(FBDD)的文库。在这 续期申请，这些MBP片段的药物样特征将通过应用 等张力替换术。这有望产生用于鉴定金属酶的新的化学物质。 缓蚀剂，同时获得更广泛的物理化学性质(例如，酸度、亲脂性) 脚手架。这些金属结合等位体(Mbi)将被用来改进一类高活性的 在上一个项目期间开发的针对流感病毒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内切酶有活性，但摄取性能较差 这些抑制剂中的一种导致了在细胞中抗病毒的活性不佳。将使用MBIS来改进 物理化学性质，同时保持基于酶的活性，以产生高活性的抑制剂 在活细胞中对抗病毒。最后，检测金属酶的靶向活性和选择性 我们的Mbps、MBIS和PAN抑制剂将通过细胞热位移分析(CETSA)进行检测。 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发展成为全长PAN内切酶抑制剂而得到改善。详细的蜂窝目标 使用这些方法检测金属酶抑制剂的接合数据很少；因此，这些研究将 对于阐明这些治疗性化合物的选择性和临床前景是有价值的。 之前的项目期产生了许多合作、专利披露、会议 会议记录，以及~13种出版物。此外，生物技术工作人员的熟练实习生 得到了指导，并将我们的成果转化为创业公司。我们将继续培育 合作发现最好和一流的有潜力改进的金属酶抑制剂 人类健康。总体而言，这项研究计划将继续在识别小型 分子抑制剂，用于具有挑战性的目标类别，对改善人类健康具有很大价值。