Chemical Biology of the Control of Neddylation by DCN1

DCN1 控制 Neddylation 的化学生物学

• 批准号: 10655433
• 负责人: Rodney Kiplin Guy
• 金额: $ 62.32万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655433
• 关键词: 3q26; Acetylation; Affinity; Amines; Amino Acids; Animals; Automobile Driving; Binding; Biochemical; Biological; Biological Availability; Biological Process; Biology; Brain; Bromodomain; C-terminal; CUL1 gene; CUL3 gene; CUL5 gene; Cell Nucleus; Cells; Cervical; Charge; Chemicals; Clinical; Complex; Contact Inhibition; Cullin Proteins; Cytosol; Data; Development; Disease; Disease Progression; Drug Targeting; Enzymes; Excretory function; Family member; Foundations; Generations; Genetic; Genetic Models; Goals; Growth Factor; Health; Homeostasis; Human; Hydrophobicity; Immune System Diseases; In Vitro; Ligase; Ligation; Location; Lysine; Malignant Neoplasms; Mammals; Measures; Mediating; Metabolism; Methionine; Modeling; Molecular; Mus; N-terminal; Names; Nature; Oncogenes; Oncoproteins; Oral; Pathway interactions; Pattern; Pharmaceutical Chemistry; Pharmacodynamics; Phenotype; Post-Translational Protein Processing; Process; Prostate; Proteasome Inhibitor; Protein Dynamics; Protein Isoforms; Proteins; RBX1 gene; Regulation; Reporting; Research Design; Research Methodology; Role; Sequence Homology; Side; Signal Transduction; Squamous cell carcinoma; Structure; Substrate Specificity; System; Testis; Tissues; Toxic effect; Ubiquitin; Ubiquitin Like Proteins; Ubiquitin family; Ubiquitination; antitumor drug; cancer therapy; clinically significant; combinatorial; drug discovery; improved; in vivo; inhibitor; knock-down; malignant phenotype; mouse genetics; novel therapeutics; pharmacologic; pre-clinical; receptor; response; small molecule; spatiotemporal; success; three dimensional cell culture; tool; tumor; tumor progression; ubiquitin ligase; ubiquitin-protein ligase

Abstract The long-term goal is to generate and use complimentary chemical and biological probes to study the cullin RING ubiquitin ligases (CRL’s) and understand their activation controlled by the interaction of by the Defective in Cullin Neddylation 1 (DCN1) and UBE2M proteins. Because the CRL’s ultimately control ubiquitination of many diverse proteins, thus regulating their stability, intracellular localization, and function, having spatiotemporal control over DCN-mediated CRL activity has the potential to unravel the mechanism regulating key cellular signaling networks and driving disease progression. The health relatedness of this project lies in two facts: 1) DCN1 is an oncoprotein, amplified in squamous cell carcinomas, that drives a highly malignant phenotype, and 2) CRL driven ubiquitination is a validated target in multiple diseases, particularly cancer and immune dysfunction. Therefore, inhibitors of the DCN1-UB2M interaction that are potent, selective, and bioavailable have the potential to be developed as antitumor drugs and possibly for other diseases. Ubiquitination is regulated by a highly complex, dynamic, and redundant network. Inhibitors of DCN1-UB2M will allow direct interrogation of the function of sub-portions of the network and are likely to unveil fundamental principles of the regulation ubiquitination. The generation of complementary cellular and mouse genetic models will enable independent verification of hypotheses. Finally, the DCN1-UBE2M interaction requires N-terminal acetylation of UBE2M, a common posttranslational modification controlling protein interactions. Therefore, a strategy for targeting N- terminal acetylation dependent protein interactions could be widely applicable. The research design and methods for achieving these goals involves the integrated and recursive use of structure-driven, hypothesis- based medicinal chemistry; in vitro biochemical measures of affinity and inhibitory potency; in vivo measures of compound efficacy and pharmacodynamic responses; and in vitro and in vivo measures of compound bioavailability, distribution, metabolism, excretion, and toxicity. The overall goal is to develop new complimentary chemical and biological tools to understand the regulation of the ubiquitin-like protein NEDD8, and uncover the specific role of DCN-mediated neddylation in Cullin-RING ligase substrate receptor exchange, growth factor signaling, and driving tumor progression. Our aims are: Aim 1: Improve the potency and oral exposure of our current DCN1/2 inhibitors and generate new chemical probes with sufficient potency and selectivity to enable ourselves and others to study the consequences of inhibiting this E2-E3 interaction in cells and animals. Aim 2. Investigate how the composition of cellular CUL1 and CUL3 ligases dynamically responds to environmental perturbations and the importance of DCN1/2 in this process. Aim 3: Use genetic and pharmacological approaches to study the effects of inhibiting DCN1 activity in animal tumor models.

抽象的 长期目标是生成和使用免费的化学和生物学问题来研究库林 环泛素连接酶（CRL），并理解其激活因有缺陷的相互作用而控制的 在Cullin Neddylation 1（DCN1）和UBE2M蛋白中。因为CRL最终控制着许多 各种蛋白质，因此控制其稳定性，细胞内定位和功能，具有时空 对DCN介导的CRL活性的控制有可能阐明调节钥匙细胞的机制 信号网络和推动疾病进展。该项目的健康相关性在于两个事实：1） DCN1是一种在鳞状细胞癌中扩增的癌蛋白，可驱动高度恶性表型，并且 2）CRL驱动泛素化是多种疾病（尤其是癌症和免疫）的验证靶标 功能障碍。因此，潜在的，选择性和生物利用的DCN1-UB2M相互作用的抑制剂具有 作为抗肿瘤药物开发的潜力，可能用于其他疾病。泛素化受到 一个高度复杂，动态和冗余的网络。 DCN1-UB2M的抑制剂将允许直接询问 网络子部分的功能，可能会揭示该法规的基本原则 泛素化。完整的细胞和小鼠遗传模型的产生将使独立 验证假设。最后，DCN1-UBE2M相互作用需要UBE2M的N末端乙酰化A 常见的翻译后修饰控制蛋白质相互作用。因此，针对n-的策略 末端乙酰化依赖性蛋白相互作用可能广泛适用。研究设计和 实现这些目标的方法涉及结构驱动的假设的综合和递归使用 - 基于医学化学；体外生化测量亲和力和抑制效力；体内测量 复合效率和药效反应；以及化合物的体外和体内测量 生物利用度，分布，代谢，极端和毒性。总体目标是发展新的免费 理解泛素样蛋白NEDD8的化学和生物学工具，并发现 DCN介导的Neddylation在Cullin-Ring连接酶底物接收器交换，生长因子中的特定作用 信号传导和驱动肿瘤进展。我们的目标是：目标1：提高我们的效力和口腔暴露 当前的DCN1/2抑制剂，并产生具有足够效力和选择性的新化学问题 我们自己和他人研究抑制细胞和动物中E2-E3相互作用的后果。目标2。 研究细胞CUL1和CUL3连接酶的组成如何动态响应环境 在此过程中，扰动和DCN1/2的重要性。目标3：使用遗传和药物 研究动物肿瘤模型中抑制DCN1活性的影响的方法。