Small Molecular Screen for Thymocyte Drug Target (RMI)

描述(申请人提供)：免疫调节是药物发现研究的一个主要目标，对传染病、免疫缺陷和自身免疫性疾病有影响。胸腺中的一种小分子药物靶点被认为是通过逆转免疫多样性的病理丧失来促进艾滋病和其他免疫缺陷的免疫系统重建的。在质量上得到改善的免疫重建是艾滋病--除了在联合逆转录病毒疗法或HAART期间发生的事情--对于减少对现有抗病毒疗法的依赖及其伴随的严重副作用，以及实施艾滋病毒治疗性疫苗至关重要。 靶标被指定为“孤儿”受体，因为选择性和/或有效的配体是未知的，因此其药理还没有被研究。这项建议的目标是为这个目标开发一种高通量筛选(HTS)方法，以便能够筛选大型(&gt；50,000)化合物文库。我们将使用HTS来确定一系列化合物作为潜在的线索，选择其中最好的来设计更有效的化合物，以阐明受体功能和药物开发。在最初的小规模筛选中，我们已经确定了一些合成的小分子配体作为HTS发展的阳性对照。该提案的目标是(I)开发用于识别激动剂和拮抗剂的分析条件，以及(Ii)将分析缩减到384孔板格式，以便在HTS环境中实施。

DESCRIPTION (provided by applicant): The regulation of immunity is a major goal of drug discovery research, with impacts on infectious disease, immunodeficiency, and autoimmune disease. A small molecule drug target has been partially characterized in the thymus that is predicted to enhance reconstitution of the immune system in AIDS and other immunodeficiencies by reversing the pathological loss of immunological diversity. Qualitatively improved immune reconstitution is AIDS-beyond what takes place during combined retroviral therapy or HAART-will be essential to reduce reliance on current antiviral therapies, with their attendant serious side effects, and to implement a therapeutic vaccine for HIV. The target of interest is designated as an "orphan" receptor because selective and/or potent ligands are not known and consequently its pharmacology has not been investigated. The objective of this proposal is to develop a high-throughput screening (HTS) assay for this target to enable screening of large (>50,000) compound libraries. We will use HTS to identify a range of compounds as potential leads, selecting the best of these to design more potent compounds for elucidation of receptor function and drug development. We have identified a few synthetic small molecule ligands as positive controls for HTS development in an initial, small-scale screen. The goals of the proposal are (i) to develop assay conditions for identifying both agonists and antagonists and (ii) to scale down the assay to a 384-well plate format for implementation in an HTS environment.