High Throughput Screening for Pharmacoperones of the V2 Receptor

描述（由申请人提供）：该提案是对PAR-12-058“高通量筛选（HTS）检测以发现化学探针（R01）的征求”的回应。该资助机会支持学术，非营利或商业HTS筛选机构之间的合作，这些机构具有必要的专业知识和经验，可以实施用于发现和开发小分子化学探针的HTS检测。拟议的工作将由提出药物酮概念（和测定）的实验室与另外两个在HTS方面具有丰富经验并可获得大型化学文库的实验室合作进行。我们将使用现有的、经过充分验证的高通量测定来鉴定抗利尿激素2型（V2）受体（V2R）的药酮。该检测将用于筛选约64万种化合物的文库，并将确定治疗肾源性尿囊性糖尿病（一种罕见且使人衰弱的疾病，目前尚无有效的药物治疗方法）中V2R通路错误的治疗分子。药物酮是一种靶向性的小分子，它扩散到细胞中，拯救错误折叠的蛋白质突变体并使其恢复功能。救援是基于一种新发现的机制：纠正突变体的细胞路径，否则这些突变体将保留在内质网中而无法发挥作用。由于这种类型筛选的新颖性，该项目也将作为鉴定大型化学文库中存在的药物酮的原型。因此，所提出的方法在治疗方法中识别具有显著新颖性的药物，依赖于目前在分子库检测管道中未表示的细胞机制；这项技术为使用HTS方法提供了一个尚未开发的机会。这种检测的发展是重要的和新颖的，因为有用的化学结构具有控制细胞的能力

DESCRIPTION (provided by applicant): This proposal is a response to PAR-12-058, "Solicitation of Assays for High Throughput Screening (HTS) to Discover Chemical Probes (R01)." This funding opportunity supports collaboration between academic, nonprofit, or commercial HTS screening facilities that have the requisite expertise and experience to implement an HTS-ready assay for the discovery and development of small molecule chemical probes. The work proposed will be conducted by collaboration between the laboratory that developed the concept of pharmacoperones (and the assay) and two other laboratories with extensive experience in HTS and access to large chemical libraries. We will use an existing and well-validated high throughput assay to identify pharmacoperones of the vasopressin type 2 (V2) receptor (V2R). The assay will be applied to screen a library of approximately 640,000 compounds and will identify therapeutic molecules for the treatment of misrouting of the V2R in nephrogenic diabetes insipidus, a rare and debilitating disease for which there are presently no effective drug treatments. Pharmacoperones are target-specific and small molecules that diffuse into cells, rescue misfolded protein mutants and restore them to function. Rescue is based on a newly appreciated mechanism: correcting the cellular routing of mutants that would otherwise be retained in the endoplasmic reticulum and unable to function. Because of the newness of this type of screen, this project will also serve as a prototype for the identification of pharmacoperones present in large chemical libraries. Accordingly, the proposed approach identifies drugs with a significant degree of novelty in therapeutic approach, relying on cellular mechanisms that are not currently represented in the Molecular Libraries assay pipeline; this technique offers an untapped opportunity for use of the HTS approach. Development of such assays is important and novel since useful chemical structures with the ability to control cellular trafficking may already be present in existing libraries, but have not been identified using existing screens. Preliminary data show that the assay was successfully transferred to the HTS facility and early screening results show that the proposed approach is likely to be successful in identification of hits.