Identification of Small Molecules that Influence Trafficking of the GnRH Receptor

影响 GnRH 受体运输的小分子的鉴定

• 批准号: 9193075
• 负责人: Jo Ann Janovick
• 金额: $ 48.63万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-10 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9193075
• 关键词: Address; Agonist; Animal Diseases; Animal Model; Biological; Biological Assay; Cell Culture Techniques; Cell membrane; Cells; Chemical Structure; Chemicals; Collaborations; Data; Development; Diffuse; Disease; Drug Kinetics; Endoplasmic Reticulum; Exhibits; Family; Follow-Up Studies; Funding Opportunities; G-Protein-Coupled Receptors; Gonadotropin Hormone Releasing Hormone; Gonadotropin-Releasing Hormone Receptor; Human; Hypogonadism; Hypothalamic structure; In Vitro; Individual; Laboratories; Libraries; Malignant Neoplasms; Methods; Molecular Bank; Mutation; Pharmaceutical Chemistry; Pharmaceutical Preparations; Property; Protocols documentation; Readiness; Reagent; Recording of previous events; Reproduction; Research; Research Institute; Research Personnel; Route; Screening Result; Series; Source; Structure; System; Techniques; Technology; Testing; Tetracyclines; Therapeutic; Therapeutic Agents; Toxic effect; Trans-Activators; Triage; Validation; Vendor; Work; analog; assay development; base; drug discovery; drug efficacy; drug metabolism; experience; high throughput screening; human disease; in vivo; misfolded protein; mutant; novel; programs; protein function; prototype; receptor; receptor expression; response; screening; small molecule; small molecule libraries; stable cell line; therapeutic target; trafficking

Project Summary This proposal is a resubmission in response to PAR-14-284, “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 Conn laboratory that developed the concept of pharmacoperones (and the assay) and the laboratories at Scripps Research Institute with extensive experience in HTS Drug Discovery, Medicinal Chemistry, and Drug Metabolism and Pharmacokinetics (DMPK). We will use an existing and well-validated high throughput assay to identify pharmacoperones of the gonadotropin releasing hormone (GnRH) receptor (GnRHR). 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. The efficacy of these drugs has been demonstrated both in cell cultures and in vivo. The assay will be applied to screen a library of more than 640,000 compounds and will identify therapeutic molecules for the treatment of misrouting of the GnRHR in diseases of reproduction. One such disease is hypothalamic hypogonadism (HH). HH is a family of disorders, a subset of which is caused by mutations in the GnRHR which result in retention in the endoplasmic reticulum. In addition to increasing the expression of mutants at the plasma membrane, pharmacoperones of this receptor increase plasma membrane expression of WT GnRHR. Only 50% of the synthesized human GnRHR is normally expressed at the plasma membrane and this percent can be increased by pharmacoperones. Accordingly, these drugs may be used in treatment of disorders in which this receptor is sub optimally expressed and cancers in which GnRH is a therapeutic target. 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 tractable hits.

项目概要 该提案是针对 PAR-14-284“高通量筛选 (HTS) 发现化学探针（R01）。这一融资机会支持以下机构之间的合作： 具有必要专业知识的学术、非营利或商业 HTS 筛选机构 以及实施 HTS 就绪检测以发现和开发小型药物的经验 分子化学探针。拟议的工作将由以下机构之间的合作进行： 康涅狄格州实验室开发了pharmaperones（和检测）的概念和 斯克里普斯研究所的实验室在高温超导药物发现方面拥有丰富的经验， 药物化学、药物代谢和药代动力学 (DMPK)。我们将使用一个 现有且经过充分验证的高通量测定法可识别 促性腺激素释放激素（GnRH）受体（GnRHR）。 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。救援基于新 赞赏的机制：纠正突变体的细胞路由，否则这些突变体将被 保留在内质网中，无法发挥功能。这些药物的功效有 已在细胞培养物和体内得到证实。该测定将用于筛选 超过 640,000 种化合物的库，并将鉴定治疗分子 生殖疾病中 GnRHR 错误路由的治疗。其中一种疾病是 下丘脑性腺功能减退症（HH）。 HH 是一系列疾病，其中一部分是由以下原因引起的 GnRHR 突变导致内质网滞留。此外 增加质膜突变体的表达，这 受体增加 WT GnRHR 的质膜表达。仅合成了50% 人类 GnRHR 通常在质膜上表达，这个百分比可以是 药用哌酮增加。因此，这些药物可用于治疗 该受体表达欠佳的疾病和 GnRH 为非最佳表达的癌症 治疗目标。由于此类屏幕的新颖性，该项目也将作为 用于鉴定大型化学库中存在的药效酮的原型。 因此，所提出的方法识别出具有显着新颖性的药物 治疗方法，依赖于目前尚未在研究中体现的细胞机制 分子文库检测管道；这项技术提供了一个未开发的机会来使用 高温超导方法。此类测定法的开发非常重要且新颖，因为有用的化学物质 具有控制细胞运输能力的结构可能已经存在于现有的 库，但尚未使用现有屏幕进行识别。初步数据显示， 检测已成功转移至 HTS 设施，早期筛选结果表明 所提出的方法可能会成功识别可处理的命中。