Identifying Chemical Modulators of CRF-Binding Protein and CRF Receptor Complexes

鉴定 CRF 结合蛋白和 CRF 受体复合物的化学调节剂

• 批准号: 8514550
• 负责人: Nicholas David Cosford
• 金额: $ 46.8万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8514550
• 关键词: Addictive Behavior; Address; Arts; Back; Behavioral; Binding; Biochemical; Biological; Biological Assay; Biology; CRF receptor type 2; Calcium; Cell Line; Cell surface; Cells; Central Nervous System Diseases; Chemicals; Chemistry; Chemosensitization; Chronic; Clinical; Cocaine; Cocaine Dependence; Collection; Complex; Corticotropin-Releasing Hormone; Corticotropin-Releasing Hormone Receptors; DA10; Data; Data Analyses; Development; Direct Costs; Disease; Dose; Drug Addiction; Economic Burden; Facilities and Administrative Costs; Fluorescence; Funding; Funding Mechanisms; G Protein-Coupled Receptor Genes; Genomics; Human; In Vitro; Income; Institutes; Institution; Laboratories; Lead; Libraries; Ligands; Measures; Mediating; Medical Research; Membrane; Metabolism; Midbrain structure; N-Methyl-D-Aspartate Receptors; Peptides; Permeability; Persons; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phase; Phase I Clinical Trials; Physiological; Physiological Adaptation; Powder dose form; Preclinical Testing; Process; Productivity; Property; Proteins; Public Health; Reagent; Receptor Signaling; Relapse; Research; Research Personnel; Role; Safety; Self Administration; Series; Services; Signal Pathway; Signal Transduction; Slice; Social Welfare; Societies; Stress; Structure; Structure-Activity Relationship; Substance abuse problem; System; Testing; Therapeutic; Toxic effect; Validation; Vendor; Ventral Tegmental Area; absorption; addiction; assay development; base; cheminformatics; cocaine use; corticotropin releasing factor-binding protein; data mining; disability; disorder later incidence prevention; dopaminergic neuron; drug seeking behavior; effective therapy; extracellular; health care delivery; high throughput screening; improved; in vivo; inhibitor/antagonist; innovation; meetings; multidisciplinary; new therapeutic target; novel; novel therapeutics; pharmacophore; pre-clinical; prevent; programs; public health relevance; radioligand; receptor; release of sequestered calcium ion into cytoplasm; research study; response; screening; small molecule; small molecule libraries; stable cell line; stressor; success; tool

DESCRIPTION (provided by applicant): There is accumulating scientific evidence showing that stressors enhance addictive behaviors and are a common cause of relapse to substance abuse. Corticotrophin releasing factor (CRF) is a 41-aa peptide that has been shown to induce various behavioral changes related to adaptation to stress. The CRF system, including the CRF-binding protein (CRF-BP) and the CRF receptors, CRF-R1 and CRF-R2, are thought to contribute, to the physiological adaptations that result from stress. It has also been shown that CRF interaction with CRF-BP may positively modulate CRF-R2 function and, further that when CRF binds to the CRF-BP, it modulates CRF-R2 signaling and contributes to stress-induced relapse to drug seeking. The aim of this application is to identify a chemical series of ligands and compounds that disrupt the interaction between CRF- BP and CRF-R2 to prevent relapse to drug seeking behaviors. Non-peptidyl chemical inhibitors would have advantages over CRF peptides, in terms of cell permeability, stability, and in vivo pharmacology. In the past, this has been difficult due to an inability to develop a suitable high through-put assay for screening against CRF-BP. To address this problem, we have developed an innovative and novel fluorescence based calcium assay where CRF-BP is expressed and tethered at the cell surface in a heterodimeric complex with CRF-R2. This has greatly facilitated our ability to find molecules that inhibit CRF-R2 activation in the presence and absence of CRF-BP. This innovation forms the basis of the high-throughput assay that we have optimized for chemical library screening. We propose to screen a targeted synthetic compound library using this assay and identify chemical inhibitors of CRF-BP-CRF-R2 receptor complex-mediated signaling. Two types of secondary assays will independently confirm any hits. Structure Activity Relations (SAR) and hit to lead optimization will be performed for prototypical inhibitors of the CRF-BP-CRF-R2 receptor complex leading to exploratory pharmacology and preclinical development. Altogether, these efforts will result in validated chemical probes for studying the biology of CRF-BP-CRF-R2 interaction in a variety of cellular and physiological contexts, with the view to developing new therapeutics for treatment of addiction.

描述（由申请人提供）：有越来越多的科学证据表明，压力增强成瘾行为，是药物滥用复发的常见原因。促肾上腺皮质激素释放因子（CRF）是一种由41个氨基酸组成的多肽，它能诱导与应激适应有关的各种行为变化。CRF系统，包括CRF结合蛋白（CRF-BP）和CRF受体CRF-R1和CRF-R2，被认为有助于应激引起的生理适应。还已经显示，CRF与CRF-BP的相互作用可以正向调节CRF-R2功能，并且进一步地，当CRF与CRF-BP结合时，其调节CRF-R2信号传导并有助于应激诱导的药物寻求复发。本申请的目的是鉴定一系列化学配体和化合物，其破坏CRF-BP和CRF-R2之间的相互作用以防止药物寻求行为的复发。在细胞渗透性、稳定性和体内药理学方面，非肽基化学抑制剂将具有优于CRF肽的优势。在过去，由于不能开发用于针对CRF-BP进行筛选的合适的高通量测定，这一直是困难的。为了解决这个问题，我们已经开发了一种创新的和新的基于荧光的钙测定法，其中CRF-BP表达并与CRF-R2以异二聚体复合物的形式拴系在细胞表面。 这极大地促进了我们发现在存在和不存在CRF-BP的情况下抑制CRF-R2活化的分子的能力。这一创新形成了我们为化学文库筛选优化的高通量测定的基础。我们建议使用该测定筛选靶向合成化合物库，并鉴定CRF-BP-CRF-R2受体复合物介导的信号传导的化学抑制剂。两种类型的二次检测将独立确认任何命中。将对CRF-BP-CRF-R2受体复合物的原型抑制剂进行构效关系（SAR）和靶向优化，从而进行探索性药理学和临床前开发。总之，这些努力将导致验证化学探针，用于研究在各种细胞和生理背景下CRF-BP-CRF-R2相互作用的生物学，以期开发用于治疗成瘾的新疗法。