High Throughput Screening Assay Development for Pharmacoperones

药用酮的高通量筛选试验开发

• 批准号: 8816427
• 负责人: P. MICHAEL CONN
• 金额: $ 9.55万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8816427
• 关键词: Throughput; Screening; Assay; Development; Pharmacoperones

DESCRIPTION (provided by applicant): This proposal is a response to PA-07-320, "Development of Assays for High-throughput Drug Screening (HTS) (R01)." The Program Announcement is a component of the Molecular Libraries Initiative, part of the NIH Roadmap for Medical Research, and supports "the development of innovative assays that may ultimately be adapted for automated screening" of molecular libraries. Our specific aim is to produce and validate screens for identification of drugs from molecular libraries which exert their actions by a newly appreciated therapeutic approach-namely, control of post-translational protein folding and, as a consequence, cellular trafficking and rescue of mutants. G protein coupled receptors (GPCRs) are frequently targeted in library screening, yet this approach generally relies on screens that identify agonists or antagonists and would have missed many of the drugs that will be identified in the proposed screens. Our 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 offers an untapped opportunity for use of the HTS approach. This proposal addresses one of the specific areas that the PA identifies as relevant, "assays for molecular chaperones or molecules that improve the post- translational targeting, folding or assembly of proteins, especially involving mutant proteins responsible for inborn errors of metabolism... (or) rare diseases." In the planned studies we will develop and characterize assays for pharmacological chaperones that improve the post-translational folding and targeting of two mutant GPCRs which cause human disease. The products of the proposed HTS assays will be useful for treatment of two rare diseases, nephrogenic diabetes insipidus and hypogonadotropic hypogonadism, each caused by an inborn error. These screens will also serve as prototypes for identification of other therapeutic molecules, especially those involving GPCRs. Many diseases are now understood to be caused by protein misfolding. Development of such assays is important and novel since the extensive use of agonist/antagonist screens alone means that useful chemical structures with the ability to control trafficking (without receptor agonism or antagonism) may already be present in existing libraries, but have not been identified using existing methods.

描述（由申请人提供）：该提案是对PA-07-320的回应，“开发高通量药物筛查（HTS）的测定法（R01）”。该计划公告是分子图书馆倡议的组成部分，这是NIH医学研究路线图的一部分，并支持“开发创新测定法，最终可以适应分子图书馆的自动筛选”。我们的具体目的是生产和验证筛查，以鉴定分子文库中的药物，这些药物通过新近欣赏的治疗方法（即控制后翻译后蛋白折叠的控制）来发挥其作用，并因此控制了细胞运输和对突变体的挽救。 G蛋白偶联受体（GPCR）经常针对库筛选，但是这种方法通常依赖于鉴定激动剂或拮抗剂的筛选，并且会错过许多将在拟议筛选中鉴定出来的药物。我们的方法依赖于当前在分子文库中未代表的细胞机制来确定具有很大新颖性的药物。这为使用HTS方法提供了未开发的机会。该建议介绍了PA识别为相关的特定领域之一：“分子伴侣或分子的测定方法可以改善蛋白质的转化后靶向，折叠或组装，尤其是涉及导致代谢前提的突变蛋白...（OR）罕见疾病的突变蛋白。”在计划的研究中，我们将开发和表征药理学伴侣的测定法，以改善引起人类疾病的两个突变GPCR的翻译后折叠和靶向。拟议的HTS分析的产物将有助于治疗两种罕见疾病，肾脏基因糖尿病肠和性疾病性疾病性疾病，每种疾病是由天生误差引起的。这些筛选还将用作鉴定其他治疗分子的原型，尤其是涉及GPCR的分子。现在，许多疾病被认为是由于蛋白质错误折叠引起的。这种测定的开发是重要且新颖的，因为仅仅对激动剂/拮抗剂筛选的广泛使用意味着具有控制运输能力的有用化学结构（没有受体激动或拮抗作用）可能已经存在于现有文库中，但尚未使用现有方法来识别。

项目成果

Pharmacological chaperones correct misfolded GPCRs and rescue function: protein trafficking as a therapeutic target.

药理学伴侣纠正错误折叠的 GPCR 和救援功能：蛋白质运输作为治疗靶点。

• DOI: 10.1007/978-94-007-4765-4_14
• 发表时间: 2012
• 期刊: Sub-cellular biochemistry
• 作者: Maya-Núñez,Guadalupe;Ulloa-Aguirre,Alfredo;Janovick,JoAnn;Conn,PMichael
• 通讯作者: Conn,PMichael

Assay strategies for identification of therapeutic leads that target protein trafficking.

用于鉴定靶向蛋白质运输的治疗先导化合物的测定策略。

• DOI: 10.1016/j.tips.2015.05.004
• 发表时间: 2015
• 期刊: Trends in pharmacological sciences
• 影响因子: 13.8
• 作者: Conn,PMichael;Spicer,TimothyP;Scampavia,Louis;Janovick,JoAnn
• 通讯作者: Janovick,JoAnn

Therapeutic rescue of misfolded/mistrafficked mutants: automation-friendly high-throughput assays for identification of pharmacoperone drugs of GPCRs.

错误折叠/错误运输突变体的治疗拯救：自动化友好的高通量测定，用于鉴定 GPCR 的 pharmaperone 药物。

• DOI: 10.1016/b978-0-12-391862-8.00001-6
• 发表时间: 2013
• 期刊: Methods in enzymology
• 作者: Smithson,DavidC;Janovick,JoAnn;Conn,PMichael
• 通讯作者: Conn,PMichael

Gonadotropin-releasing hormone receptor activates GTPase RhoA and inhibits cell invasion in the breast cancer cell line MDA-MB-231.

• DOI: 10.1186/1471-2407-12-550
• 发表时间: 2012-11-23
• 期刊: BMC cancer
• 影响因子: 3.8
• 作者: Aguilar-Rojas A;Huerta-Reyes M;Maya-Núñez G;Arechavaleta-Velásco F;Conn PM;Ulloa-Aguirre A;Valdés J
• 通讯作者: Valdés J