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

项目摘要 富含蛋白质的酪氨酸磷酸酶（STEP）是一种神经元特异性蛋白质酪氨酸磷酸酶（PTP）和一种 阿尔茨海默病（AD）的新治疗靶点，阿尔茨海默病是一种使人衰弱的神经退行性疾病， 目前没有治愈方法。最近的研究表明，STEP在AD和其他神经退行性疾病中过度活跃， 神经精神障碍涌现模型表明，STEP活动的增加干扰了 突触功能，并有助于这些疾病的特征性认知和行为缺陷。 在AD小鼠模型中STEP的敲除或药理学抑制降低了AD小鼠模型中的生化和代谢。 这些小鼠的认知缺陷，验证STEP作为治疗AD的新型药物靶标。但 唯一报道的具有细胞和体内活性的STEP抑制剂是苯并噻庚因（TC-2153），已知其 修饰DNA，因此长期服用可能会引起不良反应。在本提案中，我们计划开发 首个STEP选择性和药物样抑制剂，用于AD小鼠模型的概念验证（POC）研究。先前 产生更多药物样STEP抑制剂的努力已经失败。这些先前的高通量筛选（HTS） 这些努力利用了简单的生化筛选测定，其中截短的STEP构建体仅包含 催化域因此，他们倾向于鉴定靶向高度保守的活性位点的化合物 对STEP没有选择性。我们已经开发了一个强大的基于蛋白质热位移（PTS）的HTS平台 该技术可以检测与384孔格式的全长STEP结合的小分子。的序列 进一步表征命中的二级分析已经到位，以及关键的合作，确保最大的 成功寻找适合POC研究的小分子的可能性， AD中基于STEP的治疗策略。在目标1中，我们将使用我们的PTS对STEP变构抑制剂进行HTS- 基于筛选平台。我们将确认和表征命中化合物，并选择最有前途的 目标2中的化学优化支架，我们将提高STEP抑制剂的效力，选择性和药物- 比如财产在目标3中，我们将在细胞模型中评估所选抑制剂的疗效和特异性。我们 总体目标是开发至少一个先导系列和一个备用系列的强效和特异性STEP抑制剂 这些探针可用于后续的测试和优化，用于体内研究。

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.