Computer-aided design and development of isoform selective inhibitors of Casein Kinase 1

酪蛋白激酶 1 异构体选择性抑制剂的计算机辅助设计和开发

基本信息

批准号：
10629703
负责人：
Jun-Yong Choi
金额：
$ 15.4万
依托单位：
QUEENS COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-22 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10629703
关键词：
Amino Acids Area Binding Binding Sites Biochemical Biological Assay Biomedical Research Cell Survival Cell physiology Cells Chemical Agents Clinical Research Complex Computational Technique Computer Analysis Computer-Aided Design Core Facility Development EIF4EBP1 gene Electrostatics Enzymes Erinaceidae Family Future Goals Human Hydrogen Bonding Hydrophobicity Individual Interdisciplinary Study Investigation Knowledge Malignant Neoplasms Mass Spectrum Analysis Mediating Methods Modernization Molecular Molecular Conformation Neoplasm Metastasis Oncogenes Patients Pharmaceutical Chemistry Phosphorylation Phosphotransferases Physiological Play Proliferating Protein Isoforms Protein Kinase Rationalization Regulation Research Research Project Grants Resolution Roentgen Rays Role Series Signal Pathway Signal Transduction Signal Transduction Pathway Sodium Chloride Specificity Structure Students Synthesis Chemistry Techniques Therapeutic Therapeutic Agents Therapeutic Effect Tissues Toxic effect Tumor Suppressor Proteins Validation beta catenin cancer cell cancer therapy casein kinase I cellular development comparative computer studies design drug candidate drug discovery experience human disease inhibitor insight interest kinase inhibitor member molecular dynamics molecular modeling nanomolar novel therapeutics pharmacologic prospective skills small molecule structural biology therapeutic candidate therapeutic development tumorigenesis

项目摘要

Project Summary Protein kinases are essential to cellular function and development, and constitutively activated kinases have been widely targeted in the development of cancer therapies. Selectivity is a critical issue in kinase therapeutic discovery and biomedical research, especially in regard to off-target effects. Since the ATP binding site are highly conserved across the kinome, most ATP-competitive kinase inhibitors suffer from specificity issues, and the development of specific kinase inhibitors remains a significant bottleneck in modern drug discovery and biomedical research. Casein kinase 1 (CK1) is involved in various cellular signal transduction pathways including Wnt/β- catenin, Hedgehog, and Hippo signaling pathways, and its mis-regulation results in various human diseases. There are six CK1 isoforms in humans such as CK1α, CK1γ1, CK1γ2, CK1γ3, CK1δ, and CK1ε, and each isoform displays individual physiological roles in cellular signal transduction. Currently, CK1 isoform-selective inhibitors are rarely developed, and most of the known CK1-specific agents have been revealed as multi-target or pan-kinase inhibitors upon further investigation. Thus, it will be a compelling future direction to develop isoform selective inhibitors of CK1 for the investigation of specific signaling mechanism of CK1 isoforms in pathophysiological conditions. The objective of this research is to develop isoform selective inhibitors of CK1 via computational analysis, synthetic chemistry, biochemical assays, and structural biology. Molecular dynamics (MD) simulations of CK1 isoforms will be performed to identify distinct features in the ATP-binding pocket of each enzyme for the design of selective inhibitors. In addition, the computational techniques and analysis methods will be applied to explain the mechanisms responsible for selective inhibition of each isoform. X-ray co-crystal structures of CK1 and our new specific inhibitors will be solved to assist further design efforts and validate hypotheses generated by computational analysis and molecular modeling. Multi-step synthesis and characterization of small molecules will be mainly conducted for the optimization of the potency and specificity of our starting agents and designed compounds. ADP-Glo biochemical assays and high-resolution Mass Spectrometry analysis will be used to study the inhibition of kinase activity of CK1 isoforms. A series of cell-based proliferation and functional assays will be performed to assess cellular potency of selective inhibitors such as MTT or CellTiter-Glo cell viability assay and 4E-BP1- and LRP6-phosphorylation analyses.

项目概要蛋白激酶对于细胞功能和发育至关重要，并且组成型激活激酶选择性是激酶的一个关键问题。治疗发现和生物医学研究，特别是 ATP 结合以来的脱靶效应。位点在整个激酶组中高度保守，大多数 ATP 竞争性激酶抑制剂都存在特异性问题，并且特定激酶抑制剂的开发仍然是现代药物的一个重要瓶颈发现和生物医学研究。酪蛋白激酶 1 (CK1) 参与多种细胞信号转导途径，包括 Wnt/β- catenin、Hedgehog 和 Hippo 信号通路及其错误调节会导致多种人类疾病。人类中有六种 CK1 同工型，例如 CK1α、CK1γ1、CK1γ2、CK1γ3、CK1δ 和 CK1ε，每种亚型目前，CK1 异构体在细胞信号转导中具有个体生理作用。抑制剂很少被开发出来，大多数已知的 CK1 特异性药物已被揭示为多靶点或泛激酶抑制剂的进一步研究因此，这将是一个引人注目的未来发展方向。 CK1 异构体选择性抑制剂，用于研究 CK1 异构体的特定信号传导机制本研究的目的是开发 CK1 的异构体选择性抑制剂。通过计算分析、合成化学、生化测定和结构生物学。将进行 CK1 同工型的分子动力学 (MD) 模拟，以确定 CK1 亚型的不同特征每种酶的 ATP 结合口袋用于设计选择性抑制剂此外，还进行了计算。将应用技术和分析方法来解释选择性抑制的机制 CK1 和我们新的特异性抑制剂的 X 射线共晶结构将得到解决，以提供帮助。进一步设计工作并验证计算分析和分子建模产生的假设。小分子的多步合成和表征将主要进行优化我们的起始药物和设计的化合物的效力和特异性。高分辨率质谱分析将用于研究CK1激酶活性的抑制将进行一系列基于细胞的增殖和功能测定来评估细胞效力。选择性抑制剂，例如 MTT 或 CellTiter-Glo 细胞活力测定以及 4E-BP1- 和 LRP6-磷酸化分析。