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

• 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
• 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

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.

项目摘要 蛋白激酶是细胞功能和发育所必需的，也是结构性激活的激酶。 已被广泛用于癌症治疗的开发。选择性是激酶的一个关键问题 治疗发现和生物医学研究，特别是在非靶点效应方面。由于ATP结合 位点在整个基因组中高度保守，大多数ATP竞争性激酶抑制剂都具有专一性 问题，以及特定的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1参与多种细胞信号转导途径，包括Wnt/β-1。 Catenin、Hedgehog和Hippo信号通路及其错误调节导致多种人类疾病。 人类有6种CK1亚型，分别为CK1α、CK1γ1、CK1γ2、CK1γ3、CK1δ和CK1ε 异构体在细胞信号转导中发挥着个体的生理作用。目前，CK1亚型具有选择性 抑制剂很少被开发出来，大多数已知的CK1特异性药物已经被发现是多靶点的 或者在进一步研究的基础上再加用泛激酶抑制剂。因此，它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异构体的特异性信号机制 病理生理条件。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、生化分析和结构生物学。 将进行CK1亚型的分子动力学(MD)模拟，以确定CK1亚型的明显特征 每种酶的ATP结合口袋，用于设计选择性抑制剂。此外，计算量 将运用技术和分析方法来解释选择性抑制的机制。 每一种异构体的。CK1和我们新的特异性抑制剂的X射线共晶结构将被解决以辅助 进一步的设计工作和验证由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是为了优化 我们的起始剂和设计的化合物的效力和专一性。ADP-Glo生化分析和 高分辨质谱学分析将用于研究对CK1激酶活性的抑制 异构体。将进行一系列基于细胞的增殖和功能分析，以评估细胞潜能。 四甲基偶氮唑盐或CellTiter-Glo细胞存活率测定和4E-BP1-和LRP6-磷酸化等选择性抑制物 分析。