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：
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
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/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

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.

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。