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

• 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
• 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

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.

代谢重塑是心脏病等疾病中的基本主题 糖尿病和癌症，细胞偏离典型的燃料利用率。丙酮酸 脱氢酶复合物（PDC）是有氧葡萄糖利用及其的守门人 下调与这些疾病非常相关。因此，最近的努力集中在 关于PDC作为治疗性的激活。目前，努力仅着重于抑制 丙酮酸脱氢酶激酶（PDK），通过磷酸化抑制PDC。然而， 丙酮酸脱氢酶磷酸酶（PDP）的激活作为治疗性 磷酸化PDC以恢复活性，被忽略了。此外，众所周知 NADH抑制了PDC活性，但努力以治疗为目标这种调节机制 被忽略了。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，并优化喹酮化合物 已被证明可以从NADH抑制中恢复PDC，作为一种新的治疗策略 代谢疾病。其次，由线粒体产生的活性氧（ROS） 电子传输链（METC）被认为可以通过PDC激活增强，但是影响 PDC作为ROS的来源以及NADH/NAD和ATP需求等特定条件的来源， 与运动一样，影响ROS的稳态尚不清楚。 ROS的位点特异性量化 生成需要在各种条件下进行详细的酶模拟才能阐明。这 该项目的目的是1）实际上筛选并实验验证PDC激活剂和2） 应用数学建模以确定PDC激活对位点特异性METC ROS的影响 一代。我们从锌数据库中获得了1500万药物样的虚拟化合物 单独筛选所有PDK同工酶（1-4）在三个已知抑制位点：lipoamide，ATP/ADP， 和PZF3以找到同工酶特异性和PAN抑制剂。此虚拟复合集也将使用 通过筛选混合晶体/计算得出的PDPC-来识别PDP激活剂 PDPR复合物。我们将在指导的结构中使用PDC（E3）的NADH结合域 方法，以识别PDC的最佳喹酮类似物。详细的酶动力学模型 PDC，TCA循环和METC将集成以模拟各种氧化态 还原应力和锻炼条件以量化位点特异性线粒体ROS 生产。我们认为，我们的提议解决了激活策略的巨大差距 PDC作为治疗性，将对PDC的影响提供定量的理解 线粒体ROS产生的活性。