Small Molecules Targeting the Mitochondrial Permeability Transition

针对线粒体通透性转变的小分子

• 批准号: 8435345
• 负责人: Dennis Neil Bourdette
• 金额: $ 3.06万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8435345
• 关键词: Affinity Chromatography; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Biological; Biological Assay; Brain; Castes; Cell Death; Cells; Chemicals; Chemistry; Collaborations; Collagen Type VI; Disease; Exhibits; Future; Genomics; Goals; Heart; Human; In Situ; In Vitro; Inner mitochondrial membrane; Ion Channel; Laboratories; Letters; Medical Research; Mitochondria; Modeling; Molecular; Molecular Bank; Molecular Structure; Morphologic artifacts; Multiple Sclerosis; Mus; Muscular Dystrophies; Myocardial Infarction; Nature; Organelles; Outcome; Pathology; Pathway interactions; Permeability; Pharmaceutical Preparations; Physiological; Play; Process; Production; Program Development; Property; Proteins; Publications; Regulation; Reperfusion Injury; Research Institute; Research Personnel; Resistance; Resources; Staging; Stroke; Structure; Therapeutic; United States National Institutes of Health; base; clinically significant; contextual factors; design; high throughput screening; human disease; improved; in vitro Assay; inhibitor/antagonist; insight; mitochondrial dysfunction; mitochondrial permeability transition pore; novel; novel strategies; novel therapeutics; repository; screening; small molecule; stem; therapeutic target

DESCRIPTION (provided by applicant): Activation of the mitochondrial permeability transition pore (PTP) clearly plays a key role some of the most wide-spread and therapeutically challenging human diseases. Our studies have established that the PTP operates in two modes 1) transiently, whereby the PTP acts as a mitochondrial Ca2+ release channel or 2) persistently, which ultimately results in the cell death and disease. Although well characterized on a functional level, we have no small molecules that specifically target the PTP or the transition from transient to persistent - from normal to pathological. As a result, our goal in this applicatin is to use the resource available in the MLPCN network is to identify the probes that uniquely target the PTP. This information is critical if we are to be able to effectively identify and/or design valuable therapeutics targeting the transition of the PTP from normal to pathological. The specific objectives of this application are based in the synergy possible through the unique combination of novel approaches available in our three laboratories; Aim 1 - We will screen the available NIH SMR to identify small molecule probes able to inhibit PTP opening using a simple in vitro assay that has already been adapted to the 1536 plate format to allow screening in high throughput (HTS) formats. Aim 2 - Since the primary screen is designed to "caste a wide net", secondary screens have been developed that can also be used in HTS formats to limit to our future studies to molecules that specifically target the PTP. Aim 3 - We will initiate studies on te mechanism of action of active compounds based on assays of mitochondrial function as assessed in an in situ, whole cell context. These tertiary screens will also serve as a mechanism to assess chemically modified active compounds in an attempt to improve their biological activity. These studies will set the stage for future interrogation aimed at extending our understanding of mitochondria and PTP activity in physiological and pathological settings. Clearly, these outcomes will be fundamental to developing novel therapeutic strategies specifically targeting the pore in the many disease processes in which the PTP has been clearly implicated.

描述（由申请人提供）：线粒体渗透性转换孔（PTP）的激活显然在一些最广泛和治疗上最具挑战性的人类疾病中起着关键作用。我们的研究已经确定PTP以两种模式运作：1）瞬时性，PTP充当线粒体Ca 2+释放通道;或2）持续性，最终导致细胞死亡和疾病。虽然在功能水平上得到了很好的表征，但我们没有特异性靶向PTP或从短暂到持续-从正常到病理的转变的小分子。因此，我们在这个应用程序中目标是使用MLPCN网络中可用资源来识别唯一针对PTP的探测。如果我们要能够有效地识别和/或设计有价值的治疗靶向PTP从正常到病理的转变，则该信息是至关重要的。本申请的具体目标是基于通过我们三个实验室中可用的新方法的独特组合可能产生的协同作用;目标1 -我们将筛选可用的NIH SMR，以使用简单的体外测定来鉴定能够抑制PTP开放的小分子探针，该体外测定已经适应于1536板格式，以允许以高通量（HTS）格式进行筛选。目标2 -由于初级筛选被设计为“广撒网”，因此已经开发了次级筛选，其也可以用于HTS形式，以将我们未来的研究限制在特异性靶向PTP的分子上。目的3 -我们将启动对活性化合物的作用机制的研究，其基于在原位、全细胞背景下评估的线粒体功能测定。这些三级筛选也将作为一种机制，以评估化学修饰的活性化合物，试图提高其生物活性。这些研究将为未来的审讯奠定基础，旨在扩大我们对线粒体和PTP活性在生理和病理环境中的理解。显然，这些结果将是根本的发展新的治疗策略，特别是针对孔在许多疾病的过程中，PTP已明确牵连。