High Throughput Screening for Pharmacoperones of the V2 Receptor

V2 受体药用酮的高通量筛选

• 批准号: 8696856
• 负责人: P. MICHAEL CONN
• 金额: $ 53.74万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8696856
• 关键词: Agonist; Biological Assay; Cell Line; Cells; Chemical Structure; Chemicals; Collaborations; Data; Development; Diffuse; Disease; Endoplasmic Reticulum; Funding Opportunities; G-Protein-Coupled Receptors; Laboratories; Libraries; Methods; Molecular Bank; Nephrogenic Diabetes Insipidus; Pharmaceutical Preparations; Property; Protocols documentation; Reagent; Recording of previous events; Research; Research Personnel; Screening Result; Series; Source; Structure; Techniques; Technology; Testing; Tetracyclines; Therapeutic; Therapeutic Agents; Toxic effect; Trans-Activators; V2 Receptors; Validation; Vasopressins; Vendor; Work; analog; assay development; base; drug discovery; experience; follow-up; high throughput screening; miniaturize; mutant; novel; programs; protein function; protein misfolding; prototype; public health relevance; receptor; receptor expression; response; screening; small molecule; small molecule libraries; trafficking

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.

描述（由申请人提供）：该提案是对PAR-12-058的回应，“对高通量筛选（HTS）进行测定以发现化学探针（R01）。”这种资金机会支持学术，非营利组织或商业HTS筛查设施之间的合作，这些设施具有必要的专业知识和经验，以实施HTS就绪的测定法，以发现和开发小分子化学探针。提出的工作将通过实验室之间的合作进行，该实验室开发了Pharmacoperones（和测定法）的概念和其他两个在HTS方面具有丰富经验并获得大型化学图书馆的实验室。我们将使用现有且经过充分验证的高通量测定法来识别加压素2型（V2）受体（V2R）的药物蛋白酶。该测定法将用于筛选大约640,000种化合物的库，并将鉴定出治疗肾脏糖尿病静脉内V2R误差的治疗分子，这是一种罕见且令人衰弱的疾病，目前没有有效的药物治疗。药物蛋白酶是靶标特异性和小分子，可扩散到细胞中，营救错误折叠的蛋白质突变体并恢复其功能。救援是基于一种新欣赏的机制：纠正突变体的细胞路由，否则这些突变体将保留在内质网中，无法发挥作用。由于这种类型的屏幕的新颖性，该项目还将作为鉴定大型化学文库中存在的药物蛋白酶的原型。因此，所提出的方法在治疗方法中鉴定出具有很高新颖性的药物，依赖于当前在分子文库中尚未表示的细胞机制测定管道。该技术为使用HTS方法提供了未开发的机会。这种测定的开发非常重要且新颖，因为有用的化学结构能够控制细胞 现有图书馆可能已经存在贩运，但尚未使用现有屏幕识别。初步数据表明，该测定法已成功地转移到HTS设施，并提早筛选结果表明，所提出的方法可能在识别命中率方面成功。