Neutron encoded activity based probes

基于中子编码活动的探针

• 批准号: 10241549
• 负责人: Damian Winston Young
• 金额: $ 43.05万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10241549
• 关键词: Amino Acids; Area; Bar Codes; Basic Science; Binding; Biochemical Pathway; Biological; Catalogs; Cell Line; Cells; Chemicals; Chromatin; Clinical; Collection; Data Analyses; Defect; Detection; Development; Disease; Drug Design; Drug Targeting; Elements; Enzymes; Generations; Histone Acetylation; Histone Deacetylase; Histone Deacetylase Inhibitor; Isotopes; Lead; Libraries; Life; Ligation; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Metabolic; Methodology; Methods; Molecular Mechanisms of Action; Neutrons; Nuclear; Pattern; Pharmaceutical Preparations; Pharmacology; Photoaffinity Labels; Physiological; Piperazines; Process; Proteins; Proteome; Reaction; Reagent; Regulation; Resistance; Resolution; Sampling; Savings; Site; Specificity; Statistical Data Interpretation; Structure; Support Groups; System; Systems Analysis; Technology; Testing; Toxic effect; Treatment Efficacy; Work; base; blind; candidate selection; clinical application; combinatorial; design; drug candidate; drug development; drug discovery; high throughput screening; in vivo; inhibitor/antagonist; innovation; novel; novel drug class; scaffold; side effect; small molecule; small molecule inhibitor; success

Drugs are designed to modulate the function of a critical protein involved in a disease; however, almost all small molecules have off-targets which can mitigate or altogether terminate their therapeutic efficacy. Unfortunately, there are no general methods that can identify all the protein targets that a drug binds to in an unbiased manner. For example, Histone Deacetylase Inhibitors (HDACIs) show promising clinical activity in many diseases; however, they are generally of low to moderate specificity and may act in part through, or be hindered by, uncharacterized off-target interactions. We propose a strategy that will allow for rapid and deep pharmacological profiling of early drug candidates that 1) identifies protein targets with extraordinary confidence, 2) localizes the precise site of interaction often with amino acid residue specificity, 3) is robust against metabolic alterations, 4) distinguishes the pharmacology of metabolites, 5) quantitates differences in pharmacological activity between cellular contexts. With this approach we will quantitate all interactions that each inhibitor has with proteins in a living cell. An neutron-encoded ‘bar code’ is added to activity-based probes during a click capture and release that allows us to blindly trace the drug in a nominal mass independent manner and simultaneously introduce quantitation channels. The barcodes are revealed in the isotopic fine structure by high resolution mass spectrometry yet do not compromise sensitivity at lower resolution fragmentation spectra. The neutron bar code is implemented by moving neutrons between elements and results in a prescribed pattern of relativistic nuclear mass defects embedded in the framework of a small molecule. With this method we can confidently retrieve drug-protein reaction products from in vivo systems regardless of metabolic alterations to the HDAC inhibitors, measure a pharmacological profile and determine molecular mechanism of action. Our central hypothesis is that neutron encoded activity based probe pharmacological profiling combined with innovative fragment-based discovery to generate selective HDACIs will enable the generation of a library of highly characterized and diversely selective HDACI probes. This will be realized through three specific aims: in aim 1 we will employ a novel and systematically diverse group of sp3-enriched fragments to generate HDACIs that sample the rim region of HDACs leading to highly selective interactions. In aim 2 we will measure the pharmacological profile of our novel HDACi’s and their effect on histone acetylation. In aim 3, we will develop a multiplexed barcoding system which to enable higher detection and resolution of drug targets over the entire proteome. This overcomes many challenges universal to early stage drug development efforts that have frustrated the development of specific HDAC inhibitors in particular. Although HDACIs will be the general focus of this proposal our method is general for drug development in any area.

药物旨在调节与疾病有关的关键蛋白质的功能。但是，几乎全部很小 分子具有脱靶，可以减轻或完全终止其治疗效率。很遗憾， 没有一般方法可以识别药物以公正方式与之结合的所有蛋白质靶标。 例如，组蛋白脱乙酰基酶抑制剂（HDACIS）在许多疾病中显示出有希望的临床活性。 但是，它们通常具有低至中等的特异性，可能部分地采取行动，或者受到，或受到的阻碍 未表征的脱靶相互作用。我们提出了一种策略，该策略将允许快速而深刻 早期候选药物的药理分析1）以非凡的方式识别蛋白质靶标 置信度2）将通常具有氨基酸保留特异性的精确相互作用位置，3） 代谢改变，4）区分代谢物的药理学，5）量化 细胞环境之间的药理活性。通过这种方法，我们将量化所有互动 每种抑制剂都有活细胞中的蛋白质。中子编码的“条形码”被添加到基于活动的 点击捕获和释放期间的问题，使我们能够盲目地以名义质量独立地追踪该药物 方式并简单地引入定量渠道。条形码在同位素罚中显示 通过高分辨率质谱法进行结构，但在较低分辨率下不会损害灵敏度 碎片光谱。中子条代码是通过在元素和结果之间移动中子来实现的 以相对论核质量缺陷的规定模式嵌入了小分子的框架中。和 这种方法我们可以自信地从体内系统中检索药物蛋白质反应产物，而不管 对HDAC抑制剂的代谢改变，测量药物剖面并确定分子 作用机理。我们的中心假设是基于中子的基于活性的探针药物 分析结合创新的基于基于片段的发现以产生选择性HDACIS将使 产生高度特征和多样性的HDACI问题的库。这将通过 三个特定的目标：在AIM 1中，我们将采用一个新颖且系统地多样化的SP3增益片段 为了生成HDACI，以采样HDAC的RIM区域，从而导致高度选择性相互作用。在目标2中我们 将测量我们新型HDACI的药物谱及其对组蛋白乙酰化的影响。在AIM 3中， 我们将开发一个多路复用的条形码系统，以更高的检测和解决药物靶标的 在整个蛋白质组上。这克服了许多普遍到早期药物开发的挑战 尤其是特定HDAC抑制剂的发展的努力。虽然hdacis 将是该提案的一般重点，我们的方法是任何领域的药物开发的一般方法。