Targeting the IKK-Binding Domain of NEMO for Inhibitors Discovery

摘要 NF-κB必需调节剂（NEMO）是激活NF-κ B的必需组分。 通过募集κB激酶抑制剂IKKα发挥其功能 和IKKβ与IKK复合物的结合。抑制IKKα和IKKβ激酶之间的相互作用 而NEMO被认为是抑制NF-κB的有吸引力的治疗途径，这是由于 途径在人类疾病中的作用，包括炎症和自身免疫性疾病， 癌该策略被证明是有效的，其利用了对应于 IKKs的NEMO结合结构域（NBD），作为IKK复合物抑制剂的体外和体内研究 疾病的模型。该项目的总体目标是了解 NEMO抑制作用及其对小分子抑制剂的开发。很长的- 该项目的长期期望是建立缓蚀剂设计的结构基础， 需要高亲和力结合和特异性。该项目将通过三个 具体目标：1.将利用结构和计算方法来表征 NEMO-抑制剂复合物的结构，它依赖于IKK结合的新结构， NEMO域，容易产生X射线结构。此外，现有的结构 封闭和半开放结构的未结合NEMO将用于抑制剂设计。2.的 NEMO结构将指导肽和小分子抑制剂的合理设计， 发展成为具有与NBD不同的物理和化学性质的强效抑制剂， 测试不同结合位点的抑制潜力。3. NEMO之间的互动 和抑制剂将通过生物化学、生物物理学和细胞方法表征， 定量结合亲和力，验证结合，鉴定结合位点，并表征 装订方式和要求。此外，在细胞模型系统中的生物学评价 依赖于NF-κB信号传导的癌症的存活率，将提供以下的基本原理和理由： 未来的临床前开发和临床研究。相关研究者的专业知识 在研究相匹配的多学科性质的项目，与经验，分析 开发和生物物理表征，基于NMR的筛选和结构 测定、X射线晶体学、肽化学和合成药物化学， 癌症生物学和抗癌药物，以及在NEMO表征方面的长期经验。 这项研究的结果将进一步加深我们对需求的理解， NEMO抑制的后果，并开辟了新疗法的发展道路， 人类疾病

Abstract The NF-κB essential modulator (NEMO) is an essential component in the activation of the canonical NF-κB pathway and exerts its function by recruiting the inhibitor of κB kinases IKKα and IKKβ to the IKK complex. Inhibition of the interaction between the IKKα and IKKβ kinases and NEMO is considered an attractive therapeutic avenue to the inhibition of NF-κB, due to the pathway’s role in human diseases, encompassing inflammatory and autoimmune diseases and cancer. The strategy was shown to be effective utilizing a peptide, corresponding to the NEMO-binding domain of the IKKs (NBD), as an IKK complex inhibitor in in vitro and in vivo models of disease. The overall goal of the project is the understanding of the mechanism for NEMO inhibition and its exploitation for the development of small molecule inhibitors. The long- term expectation of the project is to establish the structural basis for inhibitor design and the requirements for high affinity binding and specificity. The project will evolve through three specific aims: 1. Structural and computational methods will be utilized to characterize the structure of NEMO-inhibitor complexes, which rely on a new construct of the IKK-binding domain of NEMO which easily yields X-ray structures. Furthermore, the available structures of unbound NEMO in a closed and semi-open structure will be utilized for inhibitor design. 2. The NEMO structures will guide the rational design of peptide and small molecule inhibitors, to develop into potent inhibitors with distinct physical and chemical properties from the NBD and testing the potential of different binding sites for inhibition. 3. The interaction between NEMO and inhibitors will be characterized by biochemical, biophysical and cellular methods, to quantify the binding affinity, validate binding, identify the binding site and characterize the binding mode and requirements. In addition, the biological evaluation in cellular model systems of cancer that rely on NF-κB signaling for survival, will provide the rationale and justification for future preclinical development and clinical studies. The expertise of the Investigators involved in the research matches the multidisciplinary character of the project, with experience in assay development and biophysical characterization, NMR-based screening and structure determination, X-ray crystallography, peptide chemistry and synthetic medicinal chemistry, cancer biology and anticancer drugs, and a long experience in the characterization of NEMO. The outcome of this research will further our understanding of the requirements and consequences of NEMO inhibition and open the way to the development of new therapies for human disease.