Small Molecules Targeting the Mitochondrial Permeability Transition

描述（由申请人提供）：线粒体通透性过渡孔（PTP）的激活显然在一些最广泛传播和治疗上具有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)瞬时运作，即PTP作为线粒体Ca2+释放通道；2)持续运作，最终导致细胞死亡和疾病。虽然在功能水平上有很好的特征，但我们没有专门针对PTP或从短暂到持续-从正常到病理转变的小分子。因此，我们在这个应用程序中的目标是使用MLPCN网络中可用的资源来识别唯一针对PTP的探测。如果我们能够有效地识别和/或设计针对PTP从正常到病理转变的有价值的治疗方法，这些信息是至关重要的。该应用程序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，以识别能够抑制PTP打开的小分子探针，使用一种简单的体外试验，该试验已经适应了1536板格式，以便在高通量（HTS）格式下进行筛选。目标2 -由于主要筛选被设计为“撒下一张大网”，二级筛选已经被开发出来，也可以用于HTS格式，以限制我们未来对特异性靶向PTP的分子的研究。目标3：我们将启动基于线粒体功能测定的活性化合物的作用机制的研究，并在原位全细胞环境中进行评估。这些三级筛选也将作为一种机制来评估化学修饰的活性化合物，以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，这些结果将是开发新的治疗策略的基础，特别是针对许多疾病过程中的孔，其中PTP已明确涉及。

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.