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 蛋白激酶对于细胞功能和发育至关重要，并且是组成型激活的激酶 已广泛针对癌症疗法的针对性。选择性是激酶中的关键问题 治疗发现和生物医学研究，尤其是在脱靶效应方面。由于ATP结合 位点在整个Kinome中高度保守，大多数ATP竞争激酶抑制剂都具有特异性 问题，并且特定激酶抑制剂的发展仍然是现代药物的重要瓶颈 发现与生物医学研究。 酪蛋白激酶1（CK1）参与了包括Wnt/β-在内的各种细胞信号转导途径 Catenin，Hedgehog和Hippo信号通路及其错误调节导致各种人类疾病。 人类中有六种CK1同工型，例如CK1α，CK1γ1，CK1γ2，CK1γ3，CK1δ和CK1ε和CK1ε，每个人都有 同工型在细胞信号转导中显示各个物理作用。目前，CK1同工型选择性 抑制剂很少开发，大多数已知的CK1特异性剂已被揭示为多目标 或进一步研究后的泛激酶抑制剂。那将是一个令人信服的未来方向 CK1的同工型选择性抑制剂用于投资CK1同工型在 病理生理状况。这项研究的目的是开发CK1的同工型选择性抑制剂 通过计算分析，合成化学，生化测定和结构生物学。 将执行CK1同工型的分子动力学（MD）模拟，以识别不同的特征 每个酶的ATP结合口袋，用于设计选择性抑制剂。另外，计算 技术和分析方法将用于解释负责选择性抑制的机制 每个同工型。 CK1的X射线共晶结构和我们的新特异性抑制剂将得到解决以帮助 进一步的设计工作和验证通过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用于优化 我们的起始代理和设计化合物的效力和特异性。 ADP-GLO生化测定法和 高分辨率质谱分析将用于研究CK1激酶活性的抑制 同工型。将进行一系列基于细胞的增殖和功能评估以评估细胞效力 选择性抑制剂，例如MTT或CellTiter-GLO细胞活力测定法和4E-BP1和LRP6-磷酸化 分析。