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

基本信息

批准号：
8486038
负责人：
Nicholas David Cosford
金额：
$ 48.75万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
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/8486038
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

项目摘要

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）：越来越多的科学证据表明，压力源会增强成瘾行为，并且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 41 个氨基酸的肽，已被证明可诱导与适应压力相关的各种行为变化。 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。研究还表明，CRF 与 CRF-BP 的相互作用可能正向调节 CRF-R2 功能，并且进一步表明，当 CRF 与 CRF-BP 结合时，它会调节 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，这些配体和化合物会破坏 CRF-BP 和 CRF-R2 之间的相互作用，以防止药物寻求行为复发。非肽基化学抑制剂在细胞通透性、稳定性和体内药理学方面比 CRF 肽具有优势。过去，由于无法开发合适的高通量检测方法来筛选 CRF-BP，这一直很困难。为了解决这个问题，我们开发了一种基于荧光的创新型钙测定法，其中 CRF-BP 在细胞表面表达并与 CRF-R2 形成异二聚体复合物。这极大地促进了我们寻找在存在和不存在 CRF-BP 的情况下抑制 CRF-R2 激活的分子的能力。这项创新构成了我们针对化学库筛选进行优化的高通量测定的基础。我们建议使用该测定法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结构活性关系 (SAR) 和先导化合物优化，从而进行探索性药理学和临床前开发。总而言之，这些努力将产生经过验证的化学探针，用于研究各种细胞和生理环境中 CRF-BP-CRF-R2 相互作用的生物学，以期开发治疗成瘾的新疗法。公共健康相关性：我们开发了一项多学科合作研究计划，生产一系列化学配体和化合物，抑制 CRF-BP-CRF-R2 受体复合物，这是治疗成瘾的新型治疗靶点。我们建议开发研究工具来探索 CRF-BP-CRF-R2 受体复合物在成瘾发展中的功能。我们将结合新颖的基于细胞的测定、高通量筛选、结构活性关系分析、基于化学的“先导化合物”优化来生成新颖配体和化合物的靶向库。该应用将促进针对 CRF-BP-CRF-R2 受体复合物的药物的开发，用于治疗成瘾。