Pyruvate Dehydrogenase Complex Activation as a Strategy to Ameliorate Metabolic Disease

丙酮酸脱氢酶复合物激活作为改善代谢疾病的策略

基本信息

批准号：
10795189
负责人：
Michael A Moxley
金额：
$ 40.28万
依托单位：
UNIVERSITY OF NEBRASKA KEARNEY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-20 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10795189
关键词：
Address Aerobic Binding Binding Sites Bioenergetics Biological Assay Cells Characteristics Citric Acid Cycle Coenzyme Q10 Complex Computers Databases Disease Docking Down-Regulation Drug Targeting Electron Transport Enzyme Kinetics Enzymes Exercise Gatekeeping Generations Glucose Glycolysis Health Heart Diseases Homeostasis Hybrids In Vitro Individual Isoenzymes Knowledge Libraries Malignant Neoplasms Membrane Potentials Metabolic Metabolic Diseases Metabolism Methods Mitochondria Modeling Monitor Multienzyme Complexes NADH Non-Insulin-Dependent Diabetes Mellitus PDH kinase PDPK1 gene Peripheral Pharmaceutical Chemistry Pharmaceutical Preparations Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Physiological Production Protein Dephosphorylation Pyruvate Dehydrogenase (Lipoamide)-Phosphatase Pyruvate Dehydrogenase Complex Quinones Reaction Reactive Oxygen Species Research Site Source Spermine Stress Structure Superoxides Therapeutic Time Tissues Ubiquinone Validation Zinc Compounds analog attenuation computational chemistry drug candidate drug-like compound glucose metabolism inhibitor insight interest kinase inhibitor kinetic model lipoamide mathematical model novel novel therapeutic intervention novel therapeutics para-benzoquinone screening simulation small molecule therapeutic target virtual virtual experiments virtual library virtual screening

项目摘要

Metabolic remodeling is an underlying theme in diseases such as heart disease, type 2 diabetes, and cancer, where cells deviate from their typical fuel utilization profile. The pyruvate dehydrogenase complex (PDC) is the gatekeeper for aerobic glucose utilization and its downregulation is greatly associated with these diseases. As such, recent efforts have focused on the activation of PDC as a therapeutic. Currently, efforts have focused only on inhibition of the pyruvate dehydrogenase kinase (PDK), which inhibits PDC by phosphorylation. However, activation of the pyruvate dehydrogenase phosphatase (PDP) as a therapeutic, which de- phosphorylates PDC to recover activity, has been ignored. Furthermore, it is well-known that PDC activity is inhibited by NADH but efforts to therapeutically target this regulatory mechanism have been overlooked. As a more comprehensive PDC activation strategy, we look to expand PDK inhibitors, identify PDP small molecule activators, and optimize quinone compounds that have been shown to recover PDC from NADH inhibition as a new therapeutic strategy for metabolic disease. Secondly, reactive oxygen species (ROS) generated from the mitochondrial electron transport chain (mETC) are thought to be enhanced by PDC activation but the impact of PDC as a source of ROS and how specific conditions such as NADH/NAD and ATP demand, as in exercise, influence ROS homeostasis is unclear. Site-specific quantification of ROS generation requires detailed enzymatic simulations in various conditions to be elucidated. The aims of this project are to 1) virtually screen and experimentally validate PDC activators and 2) apply mathematical modeling to determine effects of PDC activation on site-specific mETC ROS generation. We obtained 15M drug-like virtual compounds from the ZINC database to individually screen all PDK isozymes (1-4) at three known inhibition sites: lipoamide, ATP/ADP, and pzf3 to find isozyme specific and pan inhibitors. This virtual compound set will also be used to identify PDP activators by virtually screening a hybrid crystal/computationally derived PDPc- PDPr complex. We will use the NADH binding domain of PDC (E3), in a structure guided approach, to identify optimal quinone analogs for PDC. Detailed enzyme kinetics models of PDC, TCA cycle, and mETC will be integrated to simulate various oxidative states including reductive stress and exercising conditions to quantify site-specific mitochondrial ROS production. We believe that our proposal addresses significant gaps in strategies to activate PDC as a therapeutic and will provide a quantitative understanding of the influence of PDC activity on mitochondrial ROS production.

代谢重塑是 2 型心脏病等疾病的一个基本主题糖尿病和癌症，其中细胞偏离其典型的燃料利用情况。丙酮酸脱氢酶复合物（PDC）是有氧葡萄糖利用的看门人，其下调与这些疾病密切相关。因此，最近的努力集中在激活 PDC 作为治疗剂。目前，努力仅集中于抑制丙酮酸脱氢酶激酶 (PDK)，通过磷酸化抑制 PDC。然而，激活丙酮酸脱氢酶磷酸酶（PDP）作为治疗剂，磷酸化 PDC 以恢复活性，已被忽略。此外，众所周知的是 PDC 活性被 NADH 抑制，但针对这种调节机制的治疗努力被忽视了。作为更全面的 PDC 激活策略，我们希望扩展 PDK 抑制剂、鉴定 PDP 小分子激活剂并优化醌化合物已被证明可以从 NADH 抑制中恢复 PDC，作为一种新的治疗策略代谢性疾病。其次，线粒体产生的活性氧（ROS）电子传递链 (mETC) 被认为可以通过 PDC 激活得到增强，但影响 PDC 作为 ROS 来源的作用以及 NADH/NAD 和 ATP 等特定条件的需求，与运动中一样，对 ROS 稳态的影响尚不清楚。 ROS 的位点特异性定量生成需要在各种条件下进行详细的酶模拟才能阐明。这该项目的目标是 1) 虚拟筛选和实验验证 PDC 激活剂，2) 应用数学模型确定 PDC 激活对位点特异性 mETC ROS 的影响一代。我们从 ZINC 数据库中获得了 1500 万个类药虚拟化合物在三个已知抑制位点单独筛选所有 PDK 同工酶 (1-4)：硫辛酰胺、ATP/ADP、和 pzf3 寻找同工酶特异性抑制剂和泛抑制剂。该虚拟复合组也将被使用通过虚拟筛选混合晶体/计算得出的 PDPc 来识别 PDP 激活剂 PDPr复合物。我们将在结构引导中使用 PDC (E3) 的 NADH 结合域方法，以确定 PDC 的最佳醌类似物。详细的酶动力学模型 PDC、TCA 循环和 mETC 将被集成来模拟各种氧化状态，包括还原应激和运动条件以量化位点特异性线粒体 ROS 生产。我们相信，我们的提案解决了激活战略方面的重大差距 PDC 作为一种治疗方法，将提供对 PDC 影响的定量理解线粒体 ROS 产生的活性。