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: 8107634
• 负责人: Selena E. Bartlett
• 金额: $ 39.34万
• 依托单位: ERNEST GALLO CLINIC AND RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8107634
• 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; Screening procedure; 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; 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. PUBLIC HEALTH RELEVANCE: We have developed a multidisciplinary collaborative research program to produce a chemical series of ligands and compounds inhibit the CRF-BP-CRF-R2 receptor complex a novel therapeutic target for the treatment of addiction. We propose to generate research tools for probing the function of the CRF-BP-CRF-R2 receptor complex in the development of addiction. We will combine novel cell based assays, high throughput screening, structure activity relationship analysis, chemistry-based 'hit to lead' optimization to generate a targeted library of novel ligands and compounds. This application will facilitate the development of medications that target the CRF-BP-CRF-R2 receptor complex for the treatment of addiction.

描述（由申请人提供）：存在累积的科学证据，表明压力源增强了成瘾行为，并且是滥用药物滥用的常见原因。皮质营养素释放因子（CRF）是一种41-AA肽，已证明会诱导与压力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）被认为是对压力引起的生理适应的贡献。还表明，与CRF-BP的CRF相互作用可能会正调节CRF-R2功能，进一步，当CRF与CRF-BP结合时，它会调节CRF-R2信号传导，并有助于压力引起的对药物寻求药物的复发。该应用的目的是确定一系列化学系列的配体和化合物，这些配体破坏了CRF-BP和CRF-R2之间的相互作用，以防止对吸毒行为复发。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比CRF肽具有优势。过去，由于无法开发适合CRF-BP筛查的合适的高put测定法，这很困难。为了解决这个问题，我们开发了一种创新和新颖的基于荧光的钙测定法，其中CRF-BP在与CRF-R2的异二聚体复合物中表达和束缚在细胞表面。 这极大地促进了我们在存在和不存在CRF-BP的情况下找到抑制CRF-R2激活的分子的能力。这项创新构成了我们为化学库筛选优化的高通量测定法的基础。我们建议使用此测定法筛选靶向的合成化合物库，并确定CRF-BP-CRF-R2受体复合物介导的信号传导的化学抑制剂。两种类型的次要测定将独立确认任何命中。对于CRF-BP-CRF-R2受体复合物的原型抑制剂，将对探索性药理学和临床前开发进行结构活性关系（SAR）和打击到铅优化。总的来说，这些努力将导致在多种细胞和生理环境中研究CRF-BP-CRF-R2相互作用的生物学的经过验证的化学探针，以期开发新的治疗剂来治疗成瘾。 公共卫生相关性：我们已经开发了一项多学科的合作研究计划，以生产一系列化学的配体和化合物，抑制了CRF-BP-CRF-R2受体复合物，是成瘾治疗的新型治疗靶标。我们建议生成研究工具，以探测CRF-BP-CRF-R2受体复合物在成瘾发展中的功能。我们将结合基于新颖的细胞测定，高吞吐量筛选，结构活动关系分析，基于化学的“命中率”优化，以生成针对性的新型配体和化合物的靶向库。该应用将促进针对CRF-BP-CRF-R2受体复合物治疗成瘾的药物的开发。

项目成果

Stress and addiction: contribution of the corticotropin releasing factor (CRF) system in neuroplasticity.

• DOI: 10.3389/fnmol.2012.00091
• 发表时间: 2012
• 期刊: Frontiers in molecular neuroscience
• 影响因子: 4.8
• 作者: Haass-Koffler CL;Bartlett SE
• 通讯作者: Bartlett SE