Discovery of 12/15-Lipoxygenase Inhibitors for Alzheimer's Disease

项目总结： 这项研究的主要目标是开发选择性和有效的人类12/15-LOX抑制剂， 它将探索炎症在阿尔茨海默病(AD)中的作用，从而产生治疗阿尔茨海默病的药物 毁灭性的人类疾病。阿尔茨海默病缺乏任何治疗方法，突显了治疗其新方法的必要性 治疗。在新的靶点中，12/15-脂氧合酶(12/15-LOX或15-LOX-1)在许多 理由。12/15-LOX在中枢神经系统中广泛表达，其蛋白水平和酶活性 阿尔茨海默病患者和临床诊断为轻度认知障碍的患者活动显著增加 损伤，提示该通路早期参与了AD的发病。我们的实验室已经证明 基因缺失或早期药物阻断12/15-LOX可防止认知障碍和 阿尔茨海默病转基因小鼠模型中类AD神经病理学的研究进展。此外，我们还有 还表明，12/15-LOX的药物阻断是有益的后病理表型 疾病已经发展起来。PD146176，第一代，非专利的12/15-LOX抑制剂，拯救学习和 记忆障碍，减少Aβ水平和沉积，促进tau清除和改善突触完整性 老龄3xTg小鼠。然而，PD146176不是一种选择性的12/15-LOX抑制剂，它对 小鼠12/15-LOX相对于人类12/15-LOX。因此，发现更具选择性的下一代 可申请专利的12/15-LOX抑制剂可能会提供更好的多效性益处。 当前提案的三个目标是首先测试我们的特征良好、获得专利、强大的 选择性12/15-LOX抑制剂ML351，对抗我们的小鼠AD模型。ML351具有优异的ADME/PK性能 因此，我们将首先使用ML351重现之前在我们实验室中发现的PD146176结果，以确认它是 可行的AD治疗对12/15-LOX。第二，我们将发现更多的候选分子 抗阿尔茨海默病的治疗进展，利用我们的检测方法为人类寻找新的选择性抑制剂 12/15-LOX。我们已经成功执行了12/15-LOX高通量(HTP)筛查 与美国国立卫生研究院合作建立新的500,000个化合物图书馆。我们将筛选出前1000种化合物 在此筛选中，测试体外对人12/15-LOX的最高效力/选择性打击，并确定其在体内 对培养的小鼠HT-22神经细胞的抑制作用。第三，我们将优化我们新发现的12/15- LOX抑制剂的ADME和PK特性，并确认其LOX/COX选择性。考虑到 ML351已经被证明可以穿越血脑屏障，保护中风免受损害，我们正在 相信这些研究将表明ML351可以预防AD症状，因此具有极好的 有机会成为治疗阿尔茨海默病的有效工具。

PROJECT SUMMARY: The principle goal of this research is to develop selective and potent human 12/15-LOX inhibitors, which will probe the role of inflammation in Alzheimer's disease (AD), leading to drug candidates for this devastating human disease. The lack of any therapeutic for AD highlights the need for new approaches to its treatment. Among the novel targets, 12/15-lipoxygenase (12/15-LOX or 15-LOX-1) stands out for numerous reasons. 12/15-LOX is widely expressed in the central nervous system, and its protein levels and enzymatic activity are significantly elevated in patients with AD and those with a clinical diagnosis of mild cognitive impairment, suggesting an early involvement of the pathway in AD pathogenesis. Our laboratories have shown that genetic absence or the early pharmacological blockade of 12/15-LOX prevents cognitive impairment and the development of AD-like neuropathology in transgenic mouse models of the disease. In addition, we have also shown that the 12/15-LOX pharmacological blockade is beneficial after pathological phenotype of the disease has developed. PD146176, a first-generation, off-patent 12/15-LOX inhibitor, rescues learning and memory deficits, reduces Aβ levels and deposition, facilitates tau clearance and improves synaptic integrity in aged 3xTg mice. However, PD146176 is not a selective 12/15-LOX inhibitor and it has poor activity against the mouse 12/15-LOX relative to the human 12/15-LOX. Therefore, discovering more selective next-generation 12/15-LOX inhibitors that are patentable may provide better pleiotropic benefits. The three objectives of the current proposal are to first test our well-characterized, patented, potent, selective 12/15-LOX inhibitor, ML351, against our mouse AD model. ML351 has excellent ADME/PK properties so we will first reproduce the PD146176 results found previously in our laboratory with ML351 to confirm it as a viable AD therapeutic against 12/15-LOX. Second, we will discover additional candidate molecules toward therapeutic development against AD, utilizing our assays to identify novel, selective inhibitors for the human 12/15-LOX. We have already performed a successful 12/15-LOX high-throughput (HTP) screen in collaboration with the NIH of their new 500,000 compound library. We will screen the top 1000 compounds of this screen, test the top potent/selective hits against in vitro human 12/15-LOX, and determine their in vivo effectiveness against cultured mouse HT-22 neuronal cells. Third, we will optimize our newly discovered 12/15- LOX inhibitors for their ADME and PK properties and confirm their LOX/COX selectivity. Considering that ML351 has already been shown to cross the blood-brain barrier and protect against stroke damage, we are confident these studies will show that ML351 protects against AD symptoms and thus have an excellent chance of becoming an effective therapeutic tool against AD.