Small molecule mimetics of Humanin that normalize neuronal p-Akt as novel therapeutics for AD

项目摘要/摘要 我们最近发现的口腔脑通透性小分子模拟人的生物活性(HN) 多肽提高和正常化神经元磷酸化Akt(p-Akt)水平提供了独特的机会 评估这一新方法在阿尔茨海默病(AD)中的作用。在这项建议中，我们将把我们的努力指向 优化这类新的药物，专注于筛选更多的热门药物，增强其效力，类似于药物 在AD模型中的性质、溶解性、口腔脑渗透性和对开发该新型药物的有效性 阿尔茨海默病的治疗方法。我们还将使用建模来识别用于测试的基于HN的多肽仿制药。我们的 数据表明，这些小分子HN模拟物，如HN，可以通过激活抑制神经元死亡 通过PI3/Akt通路调节gp130受体和信号转导，为原发性高血压患者提供神经保护。 海马神经元对抗N-甲基-D-天冬氨酸和A-β诱导的神经毒性。Hn是一个天生的 线粒体衍生的脑多肽，随年龄增长而减少，可能作为神经保护因子 对抗AD相关的神经毒性。HN模拟化合物2治疗海马神经元的实验研究 导致p-Akt增加，这与其观察到的神经保护作用有关。在AD患者中， 据报道，p-Akt显著下降。同样，在老年载脂蛋白E4(ApoE4)小鼠中，有一种 与年龄匹配的ApoE3小鼠相比，脑内p-Akt显着降低，提示PI3/Akt信号转导 受ApoE4的影响，ApoE4是AD的一个危险因素。PI3/Akt信号通路的激活可转录调控基因 与记忆有关，如胆碱乙酰转移酶(ChAT)和囊泡乙酰胆碱转运体(VAChT) 也可能调节与神经可塑性有关的突触后蛋白。广告是最普遍的与年龄有关的 痴呆症，目前困扰着美国540多万人。鉴于对新技术的迫切需求 阿尔茨海默病的治疗方法，这些HN模拟物可能为治疗提供有前途的候选药物 发展。在目标1中，我们计划评估小分子HN模拟物和肽模拟物对激活 Gp130和p-Akt的正常化及其对Aβ和N-甲基-D-天冬氨酸诱导的神经毒性的神经保护作用。 在目标2中，我们将使用当前的合成孔径雷达和新的对接/建模进行设计和综合活动 识别小分子模拟物、多肽和多肽模拟物的数据。我们将优化效力、类似药物的特性， 用于药效试验的溶解度和口服脑部生物利用度。AIMS 1和AIMS 2中最好的类似物/模拟多肽 将进行体外ADMET图谱和药代动力学(PK)研究，以及磷酸蛋白质组学 分析，目标3为ApoE4(Tr)：5XFAD体内有效性测试的最佳化合物排序 作为目标4的一部分的阿尔茨海默病小鼠模型。目标是确定口服可用的HN模拟物 增强/正常化大脑p-Akt水平，改善认知。就像HN本身一样，他们也可以有更广泛的 创伤性脑损伤、中风、β所致脑血管性痴呆的治疗应用 肌萎缩侧索硬化症(ALS)，并可能导致一类新的AD临床前候选者。

PROJECT SUMMARY/ABSTRACT Our recent discovery of orally brain permeable small molecules that mimic the bioactivity of Humanin (HN) peptide to enhance and normalize neuronal phospho-Akt (p-Akt) levels provides a unique opportunity for evaluating this new approach in Alzheimer's disease (AD). In this proposal, we will direct our efforts to optimize this new class of agents, focusing on screening additional hits, enhancing their potency, drug-like properties, solubility, oral brain permeability and efficacy in an AD model towards development of this novel therapeutic approach for AD. We will also use modeling to identify HN based peptidomimetics for testing. Our data show that these small molecule HN mimetics, like HN, can suppress neuronal death through its activation of the gp130 receptor and signaling via the PI3/Akt pathway and provide neuroprotection for primary hippocampal neurons against N-methyl D-aspartate (NMDA) and Aβ-induced neurotoxicity. HN is a naturally occurring mitochondrial-derived brain peptide that decreases with age and may act as a neuroprotective factor against AD-relevant neurotoxicity. Treatment of hippocampal neurons with our HN mimetic compound 2 resulted in an increase in p-Akt, and this correlated to its observed neuroprotective effects. In AD patients, a significant decrease in p-Akt has been reported. Similarly, in aged apolipoprotein E4 (ApoE4) mice, there is a significant decrease in p-Akt in the brain relative to age-matched ApoE3 mice suggesting that PI3/Akt signaling is affected by ApoE4, a risk factor in AD. Activation of PI3/Akt signaling can transcriptionally modulate genes related to memory such as choline acetyltransferase (ChAT) and vesicular acetylcholine transporter (VAChT) and may also regulate postsynaptic proteins involved in neuroplasticity. AD is the most prevalent age-related dementia, currently afflicting more than 5.4 million people in the US. Given the urgent need for new therapeutic approaches for AD, these HN mimetics could provide promising lead candidates for therapeutic development. In Aim 1, we plan to evaluate small molecule HN mimetics and peptidomimetics for activation of gp130 and normalization of p-Akt along with their neuroprotection against Aβ and NMDA induced neurotoxicity. In Aim 2, we would conduct a design and synthesis campaign using current SAR and new docking/modeling data to identify small mimetics, peptides and peptidomimetics. We will optimize potency, drug-like properties, solubility and oral brain bioavailability for efficacy testing. The best analogs/peptidomimetics from Aims 1 and 2 will undergo in vitro ADMET profiling and pharmacokinetic (PK) studies, along with phosphoproteomics analyses, in Aim 3 to prioritize the optimal compounds for in vivo efficacy testing in the ApoE4(TR):5XFAD murine model of AD as part of Aim 4. The goal is to identify orally available HN-mimetics that enhance/normalize brain p-Akt levels and improve cognition. Like HN itself, they could also have broader therapeutic applications in traumatic brain injury (TBI), stroke, Aβ-induced cerebrovascular dementia, amyotrophic lateral sclerosis (ALS) and could lead to a new class of preclinical candidates for AD.