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

基本信息

批准号：
8351780
负责人：
Nicholas Mark Levinson
金额：
$ 9万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-13 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8351780
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 DNA Sequence Rearrangement 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 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 success tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): 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, guidingthe 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. PUBLIC HEALTH RELEVANCE: Cancer is one of the primary causes of death in the developed world, and for most patients treatment revolves mainly around the use of non-selective cytotoxic drugs, in addition to radiation therapy and surgery. Recent dramatic success in the treatment of several types of cancer with kinase inhibitors has demonstrated that a different approach, which selectively targets the molecular anomaly responsible for the disease, has many advantages. This project will study the physical principles that govern the ability of drugs to selectivity target particular protein kinases, potentially leading to new treatments for cancers caused by mutated protein kinases.

描述（由申请人提供）：蛋白激酶在调节细胞生长和增殖的信号传导途径中起着核心作用，异常激酶活性有助于许多癌症的发展。最近成功地治疗特定的癌症蛋白激酶抑制剂，尤其是肺癌和慢性髓样白血病，强调了这些蛋白质在肿瘤发生中的重要性，并突出了需要其他激酶抑制剂治疗其他癌症的需求。新的激酶抑制剂的发展是具有挑战性的，因为这些小分子靶向的主要位点的激酶ATP结合位点的高序列保守使得很难获得对特定激酶具有选择性的化合物。当前的研究旨在通过采用全新的实验方法来解决这个问题，该方法利用物理化学的进步。在AIM 1中，将使用一种称为振动stark光谱的新光谱技术来构建ATP结合位点的静电图及其在该蛋白质家族的约500个成员中的变化。这些测量值将使用具有电场振动探针的激酶抑制剂进行，其中探针报告了在ATP结合位点结合时经历的静电剂。由于这些静电图与从抑制剂的角度出现ATP结合位点中的物理环境如何出现，因此它们将直接深入了解抑制剂化学结构如何影响与激酶的相互作用。这些测量中激酶之间发现的差异可以被利用以设计更多选择性药物。在AIM 2中，将通过执行大规模结合测定法来量化这种可能性，在该测定中，将揭示激酶抑制剂面板的选择性并直接与静电测量结果进行比较，以揭示静电变化如何决定选择性的方法。而选择性分析很普遍在制药行业中，与在AIM 1中确定的静电图的比较将使抑制剂选择性的物理基础首次确定，并指导开发具有新选择性概况的抑制剂的方法。在AIM 3中，将通过研究该环境如何受蛋白质基和结合水分子的动态重排的影响来完成ATP结合位点的表征。现在，蛋白激酶构成了一组主要的药理学靶标，并且这项工作将构成第一个全面的实验研究，以了解这些蛋白质的物理特性如何决定其与药物分子的相互作用。公共卫生相关性：癌症是发达国家的主要死亡原因之一，对于大多数患者而言，除了放射治疗和手术外，主要围绕使用非选择性细胞毒性药物的治疗。最近在几种类型的癌症治疗激酶抑制剂方面取得了巨大的成功，这表明，另一种方法可以选择性地靶向负责该疾病的分子异常，具有许多优势。该项目将研究控制药物选择性特定蛋白激酶能力的物理原理，这可能会导致因突变蛋白激酶引起的癌症的新疗法。