Chemical Biology of the Control of Neddylation by DCN1

DCN1 控制 Neddylation 的化学生物学

基本信息

批准号：
10198872
负责人：
Rodney Kiplin Guy
金额：
$ 63.59万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10198872
关键词：
3q26 Acetylation Affinity Amines Amino Acids Animals Automobile Driving Binding Bioavailable Biochemical Biological Biological Availability Biological Process Biology Bromodomain C-terminal CUL1 gene CUL3 gene CUL5 gene Cell Nucleus Cells 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 Lysine Malignant Neoplasms Mammals Measures Mediating Metabolism Methionine Modeling Molecular Mus N-terminal Names Nature Oncogenes Oncoproteins Oral Pathway interactions Pattern Pharmaceutical Chemistry Pharmacodynamics Pharmacology Phenotype Post-Translational Protein Processing Process Proteasome Inhibitor 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 Brain Tissues Toxic effect Ubiquitin Ubiquitin Like Proteins Ubiquitin family Ubiquitination antitumor drug base cancer therapy clinically significant combinatorial drug discovery improved in vivo inhibitor/antagonist knock-down malignant phenotype mouse genetics novel therapeutics pre-clinical receptor response small molecule spatiotemporal success three dimensional cell culture tool tumor tumor progression ubiquitin ligase ubiquitin-protein ligase

项目摘要

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并使用互补的化学和生物探针来研究剔除 RING 泛素连接酶 (CRL) 并了解其激活是由缺陷相互作用控制的 Cullin Neddylation 1 (DCN1) 和 UBE2M 蛋白中的 CRL 最终控制着许多蛋白的泛素化。不同的蛋白质，从而调节它们的稳定性、细胞内定位和功能，具有时空性对 DCN 介导的 CRL 活性的控制有可能揭示调节关键细胞的机制该项目的健康相关性在于两个事实：1) DCN1 是一种癌蛋白，在鳞状细胞癌中扩增，导致高度恶性表型，并且 2) CRL 驱动的泛素化是多种疾病（特别是癌症和免疫疾病）中经过验证的靶点因此，具有有效、选择性和生物利用度的 DCN1-UB2M 相互作用抑制剂。被开发为抗肿瘤药物以及可能用于其他疾病的潜力受到监管。 DCN1-UB2M 的高度复杂、动态和冗余网络将允许直接询问。网络子部分的功能，并可能揭示监管的基本原则互补的细胞和小鼠遗传模型的产生将使独立成为可能。最后，DCN1-UBE2M 相互作用需要 UBE2M 的 N 端乙酰化。控制蛋白质相互作用的常见翻译后修饰因此是一种针对 N- 的策略。末端乙酰化依赖性蛋白质相互作用可广泛应用。实现这些目标的方法涉及结构驱动、假设的综合和递归使用基于药物化学；亲和力的体外生化测量和体内抑制效力的测量；化合物功效和药效学反应；以及化合物的体外和体内测量总体目标是开发新的互补药物的生物利用度、分布、代谢、排泄和毒性。化学和生物工具来了解泛素样蛋白 NEDD8 的调节，并揭示 DCN 介导的 neddylation 在 Cullin-RING 连接酶底物受体交换、生长因子中的特定作用我们的目标是：目标 1：提高我们药物的效力和口服暴露。当前的 DCN1/2 抑制剂并生成具有足够效力和选择性的新化学探针，以实现我们自己和其他人研究抑制细胞和动物中这种 E2-E3 相互作用的后果。研究细胞 CUL1 和 CUL3 连接酶的组成如何动态响应环境干扰以及 DCN1/2 在此过程中的重要性目标 3：利用遗传和药理学。研究抑制动物肿瘤模型中 DCN1 活性的影响的方法。