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）：本提案是对 PAR-12-058“用于发现化学探针 (R01) 的高通量筛选 (HTS) 测定的征集”的回应。这一资助机会支持学术、非营利或商业 HTS 筛选机构之间的合作，这些机构拥有必要的专业知识和经验来实施 HTS 就绪检测，以发现和开发小分子化学探针。拟议的工作将由开发pharmaperones（和检测）概念的实验室与另外两个在高温超导方面拥有丰富经验并可访问大型化学库的实验室之间合作进行。我们将使用现有且经过充分验证的高通量测定法来鉴定 2 型加压素 (V2) 受体 (V2R) 的药用哌酮。该测定将用于筛选大约 640,000 种化合物的库，并将鉴定用于治疗肾性尿崩症中 V2R 错误路由的治疗分子，肾性尿崩症是一种罕见且令人衰弱的疾病，目前尚无有效的药物治疗方法。药用哌酮是具有靶标特异性的小分子，可以扩散到细胞中，拯救错误折叠的蛋白质突变体并使其恢复功能。拯救基于一种新近认识的机制：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。由于此类筛选的新颖性，该项目还将作为鉴定大型化学库中存在的药用哌酮的原型。因此，所提出的方法依赖于目前分子库分析管道中尚未体现的细胞机制，识别出在治疗方法中具有显着新颖性的药物；该技术为 HTS 方法的使用提供了一个尚未开发的机会。此类测定的开发非常重要且新颖，因为具有控制细胞能力的有用化学结构 现有图书馆中可能已经存在贩运行为，但尚未使用现有屏幕进行识别。初步数据表明，该检测已成功转移到 HTS 设施，早期筛选结果表明，所提出的方法很可能成功识别命中。