Development of STEP Allosteric Inhibitors as Novel Therapeutics for Alzheimer's Disease

项目总结 纹状体富含酪氨酸磷酸酶(STEP)是一种神经元特异性蛋白酪氨酸磷酸酶(PTP)，是一种 阿尔茨海默病(AD)的新治疗靶点，这是一种衰弱的神经退行性疾病， 目前还没有治愈方法。最近的研究表明，STEP在AD和其他神经退行性变和 神经精神障碍。浮现模型表明，阶跃活动的增加干扰了 突触功能，并导致这些疾病特有的认知和行为缺陷。 敲除或药物抑制阿尔茨海默病小鼠模型的STEP可降低生化和 这些小鼠的认知缺陷，验证了STEP作为治疗AD的新药物靶点。然而， 已报道的唯一具有细胞和体内活性的步进抑制剂是一种苯并五硫氨酸(TC-2153)，已知 修改DNA，因此长期给药可能会导致不良反应。在这项提案中，我们计划开发 首次在AD小鼠模型中进行概念验证(POC)研究的STEP的选择性和类药物抑制剂。上一首 生产更多类药物步骤抑制剂的努力失败了。这些先前的高通量筛选(HTS) 努力利用了简单的生化筛选分析，其步骤结构被截断，只包含 催化域。因此，他们倾向于鉴定以高度保守的活性部位为目标的化合物 并且对步骤没有选择性。我们开发了一个基于蛋白质热位移(PTS)的健壮HTS平台 可以检测到384孔格式的全长步长的小分子结合的技术。一系列 二次化验以进一步确定HITS的特征已经到位，以及关键的合作，确保最大 在寻找适合于POC研究的小分子方面成功的可能性 阿尔茨海默病的阶梯治疗策略。在目标1中，我们将使用我们的PTS-进行步骤变构抑制剂的HTS- 基于筛选平台。我们将确认和表征Hit化合物，并选择最有希望的 目标2的化学优化支架，我们将提高阶跃抑制剂的效力、选择性和药物- 就像房产一样。在目标3中，我们将在细胞模型中评估选定的抑制剂的有效性和特异性。我们的 总体目标是开发至少一个有效和特定的阶跃抑制剂的先导系列和后备系列 为体内研究的后续测试和优化做好准备的探针。

PROJECT SUMMARY STriatal-Enriched Tyrosine Phosphatase (STEP) is a neuron-specific protein tyrosine phosphatase (PTP) and a novel therapeutic target for Alzheimer's disease (AD), a debilitating neurodegenerative disorder for which currently no cure exists. Recent studies indicate that STEP is overactive in AD and other neurodegenerative and neuropsychiatric disorders. The emergent model suggests that the increase in STEP activity interferes with synaptic function and contributes to the characteristic cognitive and behavioral deficits in these diseases. Knockout or pharmacological inhibition of STEP in a mouse model of AD decreases the biochemical and cognitive deficits in these mice, validating STEP as a novel drug target for the treatment of AD. However, the only reported STEP inhibitor with cellular and in vivo activity is a benzopentathiepin (TC-2153) that is known to modify DNA, thus likely causing adverse effects when given chronically. In this proposal we plan to develop the first selective and drug-like inhibitors of STEP for proof-of-concept (POC) studies in AD mouse models. Previous efforts to generate more drug-like STEP inhibitors have failed. These prior high-throughput screening (HTS) efforts utilized simple biochemical screening assays with truncated STEP constructs that only contained the catalytic domain. Thus, they favored the identification of compounds that target the highly conserved active site and are not selective for STEP. We have developed a robust HTS platform based on protein thermal shift (PTS) technology that can detect small molecule binding to full-length STEP in 384-well format. A sequence of secondary assays to further characterize hits is in place, as well as crucial collaborations, ensuring the greatest likelihood of success in the search for small molecules that are suitable for POC studies aimed at establishing a STEP-based treatment strategy in AD. In Aim 1 we will perform HTS for STEP allosteric inhibitors using our PTS- based screening platform. We will confirm and characterize hit compounds and select the most promising scaffolds for chemical optimization in Aim 2, where we will improve STEP inhibitor potency, selectivity, and drug- like properties. In Aim 3, we will evaluate selected inhibitors in cellular models for efficacy and specificity. Our overall goal is to develop at least one lead series and one back-up series of potent and specific STEP inhibitor probes that are ready for subsequent testing and optimization for in vivo studies.