Capture the Inhibitor: Affinity Approaches to the Discovery of Secretase Leads

描述（由申请人提供）：根据淀粉样蛋白级联假说，β-淀粉样蛋白寡聚体的形成是阿尔茨海默病发展的关键因素之一。能够有效抑制A2产生从而减少聚集量的药物将使全世界数百万患者受益。最终，获得新的化学多样性，补充现有的筛选库，可能是解锁这些治疗方法的关键。因此，我们的长期目标是从海洋中发现新的类药小分子，可用于治疗阿尔茨海默病或作为生化探针，以进一步了解这种疾病的发展。该提案的目标是我们追求这一目标的下一步，是使用新的筛选方案来识别这些先导化合物，该方案将混合物中的特定化合物与观察到的生物效应快速联系起来，而不需要费力的生物测定引导分离（目标1）。对抑制剂进行严格的生物学评估将确定最佳先导药物（目标 2）。这一提议具有创新性，因为它解决了天然产物中如何以新的方式将化学结构与活性联系起来的基本障碍，并将这一解决方案应用于阿尔茨海默氏症药物发现的问题。该提案的具体目标是： 1. 使用创新方案鉴定 2-分泌酶 (BACE1) 抑制剂，该方案使用新的 LC-MS 同质亲和力测定快速将混合物中的特定化合物与观察到的生物活性联系起来。 2. 使用旨在确定最佳候选者的一系列测定来评估新抑制剂作为 BACE1 先导化合物的潜力。我们期望鉴定出结构上前所未有的 BACE1 抑制剂类别。这些新型潜在抗阿尔茨海默病先导药物将通过一系列生化测定进行评估，以评估对中枢神经系统药物开发至关重要的参数。满足这些要求的有限数量的化合物将与工业界合作进行进一步研究。如果成功，新发现的先导化合物预计将产生积极影响，最终验证 BACE1 作为药物靶点，验证淀粉样蛋白级联假说，并为阿尔茨海默病患者提供急需的治疗方法。 公共健康相关性：该提案是对海洋独特化学资源的首次系统检查，以寻找针对 BACE1 酶的先导药物，BACE1 酶是阿尔茨海默病发展的核心。为了实现这一目标，该提案利用了我们最近开发的新筛选方案，该方案旨在快速将生物活性与复杂混合物中的特定化学物质联系起来。

DESCRIPTION (provided by applicant): The formation of oligomers of amyloid-beta is one of the critical factors in the development of Alzheimer's disease, according to the Amyloid Cascade Hypothesis. Drugs that can effectively inhibit A2 production and thus reduce the amount of aggregation would benefit millions of patients world-wide. Ultimately, accessing novel chemical diversity, which complements existing screening libraries, may be the key to unlocking these treatments. Our long-term goal therefore is to discover new drug-like small molecules from marine sources that can be used for the treatment of Alzheimer's disease or as biochemical probes to further our under- standing of the development of this disorder. The objective of this proposal, which is our next step in pursuit of that goal, is to identify these lead compounds using a new screening protocol that will rapidly link a spe- cific compound in a mixture to the observed biological effect without requiring laborious bioassay guided isolation (Aim 1). Rigorous biological evaluation of the inhibitors will identify the best lead (Aim 2). This pro- posal is innovative because it addresses the fundamental barrier in natural products of how to link chemical structure to activity in a new way and applies this solution to the problem of Alzheimer's drug discovery. The specific aims of the proposal are: 1. To identify inhibitors of 2-secretase (BACE1) using an innovative protocol that rapidly links a specific compound within a mixture to the observed biological activity using a new LC-MS homogenous affinity assay. 2. To evaluate the potential of the new inhibitors as BACE1 leads using a gauntlet of assays designed to identify the best candidate. We expect to identify structurally unprecedented classes of BACE1 inhibitors. These new classes of poten- tial anti-Alzheimer drug leads will be evaluated in a series of biochemical assays to assess parameters criti- cal to CNS drug development. A limited number of compounds that meet these requirements will be ad- vanced to further studies in partnership with industry. If successful, the newly discovered leads are ex- pected to have a positive impact by finally validating BACE1 as a drug target, validating the Amyloid Cas- cade Hypothesis and providing desperately needed therapeutics for Alzheimer's patients. PUBLIC HEALTH RELEVANCE: This proposal is the first systematic examination of the ocean's unique chemical resources for drug leads that target the enzyme BACE1, which is central to the development of Alzheimer's disease. To accomplish this goal, the proposal leverages a new screening protocol that we recently developed, which is designed to rapidly link biological activity to a specific chemical in a complex mixture.