Novel Technology for Screening Receptor and Compound Libraries in Drug Discovery

描述（由申请人提供）：制药/生物技术行业中的发现研究采用高通量、无标记的药物结合细胞分析，其中许多是功能分析。在这种情况下，受体-配体相互作用仅是结合的间接测量，并且可能取决于激活途径的结合阈值。辐射监测设备公司(RMD)提出开发能够高通量筛选生物/生物活性分子与其同源受体之间的关联的平台技术和装置。该技术基于测定纳米颗粒的磁双折射弛豫，并将特别设计用于筛选痛敏肽受体和相互作用配体。RMD已经证明，用配体分子修饰的磁赤铁矿纳米颗粒在与蛋白质、病毒和小尺寸微生物结合时表现出更长的磁双折射弛豫时间。双折射的绝对大小和弛豫时间仅取决于“受体-配体复合物”形成时的流体动力学体积变化，并且不受过量未结合配体的影响。这些特性使得该检测方法非常适合用于筛选固定在磁性纳米颗粒表面上的分子对抗受体。用于证明概念验证的测定平台将利用固定在磁性纳米颗粒表面上的肽配体（痛敏肽和衍生物），其将在结合到在微生物（例如细菌和酵母）的细胞表面上过表达的痛敏肽受体后针对磁双折射弛豫的变化进行筛选。使用磁光双折射来鉴定配体和受体相互作用的基本原理是，它消除了将未结合的配体与被受体捕获的配体分离的需要。由于在检测之前不需要将样品与捕获颗粒分离，因此缩短了鉴定时间。重要的是，不需要将外源性荧光或放射性标记附接至受体或配体。此外，这种新方法适用于配体与同源受体的相互作用不激活易于用比率荧光平台监测的细胞内途径的系统。该方法/装置将可用于制药/生物技术工业中的广泛应用，包括开发小分子药物的发现研究和从展示文库中鉴定生物活性肽和蛋白质。另一个应用是孤儿受体的鉴定和感兴趣的配体的受体的发现。 公共卫生相关性：内源性神经肽配体-受体系统，即痛敏素/痛敏素受体，参与广泛的生理反应，包括乙醇消耗和药物滥用引起的奖赏，并且代表了可行的药物靶标。我们将开发一种策略和平台，用于筛选化合物库，以开发用于伤害感受素/伤害感受素受体系统的新型激动剂和拮抗剂。这种技术具有巨大的健康和社会经济影响的潜力，因为定义药物和酒精成瘾行为的潜在分子成分将提供更有效治疗成瘾的方法。

DESCRIPTION (provided by applicant): Discovery research in the pharmaceutical/biotech industries utilizes high throughput, label-free drug binding cell-based assays, many of which are functional assays. In such instances, the receptor-ligand interaction is only an indirect measurement of binding and might depend on a threshold of binding for activation of the pathway. Radiation Monitoring Devices, Inc. (RMD) proposes to develop a platform technology and device capable of high throughput screening for association between biologically/pharmacologically active molecules and their cognate receptors. This technology is based on assaying magnetic birefringence relaxation of nanoparticles and will be particularly designed for screening nociceptin receptor and interacting ligands. RMD has demonstrated that maghemite nanoparticles, decorated with ligand molecules, exhibit longer magnetic birefringence relaxation times when associated with proteins, viruses and small size microorganisms. The absolute magnitude of birefringence and the relaxation time depends only on the hydrodynamic volume change upon formation of the "receptor-ligand complex" and is not affected by excess unbound ligand. These properties make this assay ideal for screening molecules immobilized on the surface of magnetic nanoparticles against receptors. The assay platform to demonstrate the proof of concept will utilize a peptide ligand (nociceptin and derivatives) immobilized on the surface of magnetic nanoparticles, which will be screened for a change in magnetic birefringence relaxation upon binding to nociceptin receptor overexpressed on the cell surface of microorganisms such as bacteria and yeast. The rationale for using magneto-optical birefringence to identify ligand and receptor interactions is that it eliminates th need for separating the unbound ligands from ligands captured by receptors. The time to identification is reduced since there is no need to segregate sample from the capture particle before detection. Importantly, attachment of an exogenous fluorescent or radioactive label to the receptor or ligand is not required. Further, this novel approach is applicable in systems where the interaction of ligand with cognate receptor does not activate intracellular pathways that are easy to monitor with a ratiofluorometric platform. This procedure/device will be useful for a broad range of applications in the pharmaceutical/biotechnological industry including discovery research to develop small molecule drugs and the identification of biologically active peptides and proteins from display libraries. Another application is the identification of orphan receptors and discovery of receptors for ligands of interest. PUBLIC HEALTH RELEVANCE: The endogenous neuropeptide ligand-receptor system, the nociceptin/nociceptin receptor, is involved in a wide range of physiological responses including in reward elicited by ethanol consumption and drug abuse and represents a viable drug target. We will develop a strategy and platform for screening compound libraries for the development of novel agonists and antagonists for the nociceptin/nociceptin receptor system. Such technology carries the potential of great health and socioeconomic impact as the underlying molecular components that define drug and alcohol addiction behavior will provide insight into ways that addiction can be treated more effectively.