Kinome-Wide Spectroscopic Study of Drug Binding Site Electrostatics

药物结合位点静电的全激酶组光谱研究

• 批准号: 8973668
• 负责人: Nicholas Mark Levinson
• 金额: $ 24.82万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-13 至 2017-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8973668
• 关键词: Address; Affect; Benchmarking; Binding; Binding Sites; Biological Assay; Cancer Etiology; Cause of Death; Cell Proliferation; Chemical Structure; Chemicals; Chronic Myeloid Leukemia; Clinic; Comparative Study; Complex; Cytotoxic agent; Data Set; Development; Disease; Drug Binding Site; Drug Industry; Effectiveness; Electrostatics; Environment; Fluorescence; Fluorescence Spectroscopy; Fluorescent Probes; Goals; Growth; Human; Hydrogen Bonding; Location; Malignant Neoplasms; Malignant neoplasm of lung; Maps; Measurement; Measures; Mentors; Methods; Molecular Target; Mutate; Mutation; Pathology; Patients; Pharmaceutical Preparations; Phase; Phosphotransferases; Physical Chemistry; Physical environment; Play; Property; Protein Family; Protein Kinase; Protein Kinase Inhibitors; Proteins; Radiation therapy; Radiosurgery; Reporting; Research; Role; Series; Signal Pathway; Site; Spectrum Analysis; Structure; Techniques; Testing; Time; Translating; Variant; Water; Work; abstracting; analog; base; cancer type; chemical group; design; electric field; experience; inhibitor/antagonist; insight; kinase inhibitor; member; molecular dynamics; physical property; protein kinase inhibitor; research study; scaffold; small molecule; spectroscopic survey; success; tumorigenesis

Project Summary / Abstract Protein kinases play central roles in the signaling pathways that regulate the growth and proliferation of cells, and aberrant kinase activity contributes to the development of many cancers. Recent success in treating particular cancers with targeted protein kinases inhibitors, notably lung cancer and chronic myeloid leukemia, underscores the importance of these proteins in oncogenesis, and highlights the need for additional kinase inhibitors to treat other cancers. The development of new kinase inhibitors is challenging because the high sequence conservation of the kinase ATP-binding site, the major site targeted by these small molecules, makes it difficult to obtain compounds that are selective for particular kinases. The current study aims to address this problem through an entirely new experimental approach that utilizes advances in physical chemistry. In Aim 1, a new spectroscopic technique called vibrational Stark spectroscopy will be used to construct a map of the electrostatics of the ATP-binding site and how it varies across the ~500 members of this protein family. These measurements will be made using kinase inhibitors that possess vibrational probes of electric field, in which the probes report on the electrostatics they experience when bound in the ATP- binding site. Because these electrostatic maps relate to how the physical environment in the ATP- binding site appears from the perspective of the inhibitors, they will yield direct insight into how changes to the chemical structure of the inhibitors would affect the interaction with kinases. Differences uncovered between kinases in these measurements could be exploited to design more selective drugs. In Aim 2, this possibility will be quantified by performing large-scale binding assays in which the selectivity of panels of kinase inhibitors will be revealed and directly compared to the electrostatics measurements to reveal how electrostatic variation dictates selectivity. While selectivity profiling is commonplace in the pharmaceutical industry, the comparison with the electrostatic maps determined in Aim 1 will allow the physical basis of inhibitor selectivity to be determined for the first time, guiding the way to the development of inhibitors with new selectivity profiles. In Aim 3 the characterization of the ATP-binding site will be completed by studying how this environment is affected by the dynamic rearrangements of protein groups and bound water molecules. The protein kinases now constitute a major group of pharmacological targets, and taken together this work will constitute the first comprehensive experimental study of how the physical properties of these proteins dictate their interaction with drug molecules.

项目概要/摘要 蛋白激酶在调节生长和增殖的信号通路中发挥核心作用 细胞和异常的激酶活性导致许多癌症的发展。最近的 使用靶向蛋白激酶抑制剂成功治疗特定癌症，特别是肺癌和 慢性粒细胞白血病，强调这些蛋白质在肿瘤发生中的重要性，以及 强调需要额外的激酶抑制剂来治疗其他癌症。新的发展 激酶抑制剂具有挑战性，因为激酶 ATP 结合的序列高度保守 位点是这些小分子的主要靶向位点，因此很难获得 对特定激酶具有选择性。目前的研究旨在通过完全解决这个问题 利用物理化学进步的新实验方法。在目标 1 中，一个新的 称为振动斯塔克光谱的光谱技术将用于构建地图 ATP 结合位点的静电特性及其在该蛋白质约 500 个成员中的变化 家庭。这些测量将使用具有振动探针的激酶抑制剂进行 电场，其中探针报告它们在 ATP- 中结合时所经历的静电 结合位点。因为这些静电图与 ATP 中的物理环境有关 结合位点是从抑制剂的角度出现的，它们将直接洞察如何 抑制剂化学结构的改变会影响与激酶的相互作用。 这些测量中发现的激酶之间的差异可用于设计更多 选择性药物。在目标 2 中，将通过在以下环境中进行大规模结合测定来量化这种可能性： 激酶抑制剂组的选择性将被揭示并直接与 静电测量揭示静电变化如何决定选择性。同时选择性 分析在制药行业很常见，与静电图的比较 目标 1 中确定的结果将允许首先确定抑制剂选择性的物理基础 时间，指导开发具有新选择性的抑制剂。在目标 3 中 ATP 结合位点的表征将通过研究该环境的情况来完成 受蛋白质基团和结合水分子动态重排的影响。蛋白质 激酶现在构成了一组主要的药理学靶点，综合起来，这项工作将 构成了关于这些蛋白质的物理性质如何变化的第一个全面的实验研究 决定它们与药物分子的相互作用。