Identification of Novel ACTH small molecule Antagonists

新型ACTH小分子拮抗剂的鉴定

• 批准号: 6833598
• 负责人: ILYA OKUN
• 金额: $ 10万
• 依托单位: CHEMDIV, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6833598
• 关键词: adrenal disorder; adrenocorticotropic hormone; arrestins; biotechnology; computer assisted sequence analysis; computer simulation; computer system design /evaluation; cyclic AMP; drug discovery /isolation; green fluorescent proteins; hormone inhibitor; hormone receptor; microarray technology; molecular biology information system; molecular cloning; peptide library; receptor binding; small molecule; tissue /cell culture; transfection /expression vector

DESCRIPTION (provided by applicant): The production of adrenal gland hormones, including cortisol and androgens, is regulated by pituitary ACTH (adrenocorticotropin hormone), which acts via the melanocortin-2 receptor (MC2R) to stimulate steroid production. The ability to antagonize ACTH action would greatly facilitate the care of several clinical conditions, including congenital adrenal hyperplasia and Cushings Disease, which affect the adrenal gland. Yet, ACTH antagonists are not currently available. Thus the specific aim of this proposal is to discover new small molecule blockers of the MC2R with the long-term goal to develop them into drugs to treat the diseases mentioned above. For the purpose of this proposal, we will use Artificial Neural Networks and Support Vector Machines classification methods to design and synthesize targeted drug discovery library of potential ligands for peptide binding G-protein coupled receptors (GPCR), like MC2R. As a screening assay system, we have developed a dual expression sensor cell line, which simultaneously expresses MC2R and GFP-beta-Arrestin fusion protein. Upon the receptor stimulation with ACTH, the MC2R-GFP-beta-Arrestin complex migrates into endosomes. The process can be visualized and quantified with a fluorescent microscopy. In the presence of the receptor inhibition, we expect that the MC2R-GFP-beta-Arrestin complex will be prevented, which will allow for initial hit selection. The hits found will be entered into follow up investigation using both receptor binding techniques to characterize their affinities on membrane preparations containing the MC2R and functional cell responses, cAMP accumulation, to characterize their potencies and efficacies. These studies will involve the combined efforts of Chemical Diversity Labs, Inc., which has expertise in drug discovery chemistry, and a Yale University laboratory with expertise in GPCR action and the treatment of adrenal disorders.

描述(申请人提供)：肾上腺激素的产生，包括皮质醇和雄激素，由垂体ACTH(促肾上腺皮质激素)调节，它通过黑素皮质素-2受体(MC2R)刺激类固醇的产生。拮抗ACTH作用的能力将极大地促进几种临床疾病的治疗，包括影响肾上腺的先天性肾上腺增生症和库兴氏病。然而，ACTH拮抗剂目前还不可用。因此，这项建议的具体目标是发现新的MC2R小分子阻滞剂，长期目标是将它们开发成治疗上述疾病的药物。 为了达到这一目的，我们将使用人工神经网络和支持向量机分类方法来设计和合成针对多肽结合G蛋白偶联受体(GPCR)的潜在配体的靶向药物发现文库，如MC2R。作为筛选检测系统，我们建立了同时表达MC2R和GFP-β-arrestin融合蛋白的双表达传感细胞系。当ACTH刺激受体时，MC2R-GFP-β-arrestin复合体迁移到内体。这一过程可以用荧光显微镜进行可视化和量化。在受体抑制的存在下，我们预计MC2R-GFP-β-arrestin复合体将被阻止，这将允许最初的命中选择。已发现的HITS将进入后续研究，使用受体结合技术来表征它们在含有MC2R的膜制剂上的亲和力和功能细胞反应，cAMP积累，以表征它们的效力和有效性。 这些研究将涉及化学多样性实验室公司和耶鲁大学实验室的联合努力。化学多样性实验室公司在药物发现化学方面具有专业知识，耶鲁大学实验室在GPCR作用和肾上腺疾病治疗方面具有专业知识。