Inducers of Mitochondrial Biogenesis

线粒体生物发生诱导剂

• 批准号: 8200080
• 负责人: Craig Cano Beeson
• 金额: $ 31.21万
• 依托单位: MITOHEALTH, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8200080
• 关键词: Academia; Acute; Aging; Biochemical; Biochemical Markers; Biogenesis; Biological Assay; Biological Markers; Brain; Cell Death; Cell model; Cells; Chemicals; Chronic; Chronic Disease; Computer Simulation; Diabetes Mellitus; Disease; Drug Exposure; Drug Industry; Exhibits; Genus Hippocampus; Goals; Gold; Heart; Injury; Ischemia; Kidney; Lead; Libraries; Link; Liver; Measures; Metabolic syndrome; Metabolism; Mitochondria; Mitochondrial DNA; Mitochondrial Proteins; Modeling; Molecular; Mus; Natural regeneration; Nerve Degeneration; Organ; Pathway interactions; Pharmaceutical Preparations; Phase; Process; Quantitative Structure-Activity Relationship; Recovery; Reperfusion Therapy; Research; Screening procedure; Signal Pathway; Surrogate Markers; System; Testing; Time; Tissues; Toxicant exposure; Trauma; Tubular formation; Validation; analog; base; cell injury; design; drug development; effective therapy; high throughput screening; improved; in vivo; kidney cell; mitochondrial dysfunction; novel; pharmacophore; protein expression; repaired; research study; single molecule; small molecule libraries; toxicant

DESCRIPTION (provided by applicant): The long-term goal of this project is to identify and develop drugs that induce mitochondrial biogenesis. A variety of diverse acute insults such as ischemia/reperfusion (I/R), drug, toxicant, and trauma lead to mitochondrial dysfunction and results in cell injury and death in many organs (e.g. heart, brain, liver and kidney). Furthermore, mitochondrial dysfunction is a component of many chronic diseases such as metabolic syndrome and neurodegeneration, and aging. There are no effective therapies that can restore mitochondrial function and, thus, a drug that produces mitochondrial biogenesis may have extensive utility in the treatment of a broad group of diseases. Mitochondrial biogenesis drug development has been hindered by poor cellular models with limited mitochondrial function, surrogate markers of biogenesis that have high false-positive and false negative-rates, and no moderate to high throughput screening assay to measure functional mitochondrial biogenesis. We have studied mitochondrial biogenesis for the past five years in primary cultures of renal proximal tubular cells (RPTC) that maintain in vivo like mitochondrial function. We recently adapted and optimized our RPTC system to the 96-well Seahorse Biosciences respirometer and validated the assay using chemicals known to induce mitochondrial biogenesis. It was found that the maximum FCCP-uncoupled rates in these cells are limited only by the number of functional mitochondria and, thus, increases in the FCCP- uncoupled rates are functional biomarkers of mitochondrial biogenesis. Using this assay we screened a test library of 500 compounds randomly selected from the DIVERSet 50,000-compound library. We identified 7 hits that were confirmed with secondary assays of increased expression of mitochondrial proteins, the "gold standard". Thus, we can start immediately to use our phenotypic assay with compound libraries to identify new inducers of mitochondrial biogenesis. We hypothesize that screening a large, diverse chemical library with our assay will result in the identification of new chemotypes that produce mitochondrial biogenesis, leading to new chemical entities that produce mitochondrial biogenesis in vivo. In Phase I of the project we propose to screen the DIVERSet 50,000 compound library. The library will be initially screened as pools of five compounds (Aim 1) followed by deconvolution to single molecules (Aim 2), and finally validation of the hits using biochemical assays for mitochondrial proteins and DNA (Aim 3). In Phase II, we will identify chemotypes using in silico pharmacophore analyses and QSAR, synthesize analogues to improve activity, and determine efficacy and potency of the best analogues in inducing mitochondrial biogenesis in RPTC and in mice. ) PUBLIC HEALTH RELEVANCE: Mitochondrial dysfunction is a major contributor to acute tissue injuries and chronic diseases. At this time, there are no effective therapies that can restore mitochondrial function to promote cell and organ repair/regeneration after diverse acute and chronic injuries. Thus, research is needed to develop new drugs that promote mitochondrial biogenesis.

描述（由申请人提供）：本项目的长期目标是鉴定和开发诱导线粒体生物合成的药物。多种急性损伤如缺血/再灌注（I/R）、药物、毒物和创伤导致线粒体功能障碍并导致许多器官（例如心脏、脑、肝脏和肾脏）中的细胞损伤和死亡。此外，线粒体功能障碍是许多慢性疾病如代谢综合征和神经变性以及衰老的组成部分。没有可以恢复线粒体功能的有效疗法，因此，产生线粒体生物发生的药物可能在治疗广泛的疾病中具有广泛的用途。线粒体生物发生药物的开发受到以下因素的阻碍：线粒体功能有限的细胞模型不佳、具有高假阳性率和假阴性率的生物发生替代标志物、以及没有中到高通量筛选测定来测量功能性线粒体生物发生。在过去的五年中，我们研究了维持体内线粒体功能的肾近端小管细胞（RPTC）的原代培养物中的线粒体生物发生。我们最近调整和优化了我们的RPTC系统，以96孔Seahorse Biosciences呼吸计，并使用已知诱导线粒体生物合成的化学物质验证了该测定。发现这些细胞中的最大FCCP解偶联速率仅受功能性线粒体的数量限制，因此，FCCP解偶联速率的增加是线粒体生物发生的功能性生物标志物。使用该测定，我们筛选了从DIVERSet 50，000化合物库中随机选择的500种化合物的测试库。我们确定了7个命中，证实了与线粒体蛋白质的表达增加，“金标准”的二次测定。因此，我们可以立即开始使用我们的表型测定与化合物文库来鉴定线粒体生物发生的新诱导剂。我们假设，用我们的测定筛选大的、多样的化学文库将导致产生线粒体生物发生的新化学型的鉴定，从而导致在体内产生线粒体生物发生的新化学实体。在项目的第一阶段，我们建议筛选DIVERSet 50，000化合物库。文库将首先筛选为五种化合物的库（目标1），然后解卷积为单个分子（目标2），最后使用线粒体蛋白和DNA的生化测定验证命中（目标3）。在第二阶段，我们将使用计算机药效团分析和QSAR确定化学型，合成类似物以提高活性，并确定最佳类似物在诱导RPTC和小鼠线粒体生物合成中的功效和效力。) 公共卫生相关性：线粒体功能障碍是急性组织损伤和慢性疾病的主要原因。目前，还没有有效的治疗方法可以在各种急性和慢性损伤后恢复线粒体功能以促进细胞和器官修复/再生。因此，需要研究开发促进线粒体生物合成的新药。