Identification of Old-Age-Associated Alzheimer's Disease Drug Targets

描述（由申请人提供）：到目前为止，老年是大多数神经退行性疾病的最大风险因素，但很少有人尝试确定任何这些疾病（包括阿尔茨海默氏痴呆症（AD））的衰老相关治疗靶点。在过去的十年中，索尔克实验室已经开发了神经细胞培养药物筛选试验，该试验基于老年相关的CNS病理，包括营养支持的丧失、能量代谢降低、氧化损伤和细胞内聚集蛋白的积累。使用这些测定结合迭代药物化学，制备了一种特别有效的化合物，称为J147，其在所有上述测定中具有神经保护作用，EC 50在低纳摩尔范围内。这种化合物还可以增强年轻和非常老（30个月）小鼠的记忆力，改善老年（23个月）AD转基因小鼠的认知功能和突触结构，并延长果蝇的寿命。由于J147在缺乏淀粉样蛋白途径的任何公认的AD药物靶标的多种细胞培养模型中防止细胞死亡，因此它必须作用于与淀粉样蛋白途径无关并且有助于其治疗功效的高度相关的分子靶标和靶标途径。鉴于J147在多种功能不同的年龄相关毒性试验中具有神经保护作用， 因此，有可能定义共同的、共享的神经保护途径，然后将其用作鉴定治疗AD的新治疗靶点的基础。这种化学生物学方法用于识别药物靶点，起源于1785年从毛地黄中分离出地高辛，随后是1806年从罂粟中分离出吗啡，以及1897年从水杨酸中分离出阿司匹林。这些分子不仅对医学做出了巨大的贡献，它们也是分别涉及Na-K-ATP泵、阿片受体和环加氧酶的分子途径和药物靶点的++鉴定的关键因素，导致了今天在临床上的一系列衍生药物。本申请的总体目标是使用基于J147化合物的化学生物学方法来鉴定和验证用于AD治疗和预防的新型老年相关靶标。这将通过使用分子生物学和蛋白质组学来定义在多种毒性试验中导致神经保护的共享分子信号传导途径，以及化学和蛋白质组学来分离J147的分子靶标来完成。体内靶点验证将部分在动物模型中进行， 并不偏向于家族性AD/淀粉样蛋白通路，而是反映了人类衰老和AD的大多数方面，即加速衰老的SAMP 8小鼠。

DESCRIPTION (provided by applicant): Old age is by far the greatest risk factor for the majority of neurodegenerative diseases, yet few attempts have been made to identify aging-associated therapeutic targets for any of these diseases, including Alzheimer's dementia (AD). During the last ten years the Salk laboratories have developed nerve cell culture drug screening assays that are based upon old-age-associated CNS pathologies, including the loss of trophic support, reduced energy metabolism, oxidative damage, and the accumulation of intracellular aggregated proteins. Using these assays in conjunction with iterative medicinal chemistry, an exceptionally potent compound, called J147, was made that is neuroprotective in all of the above assays with EC50s in the low nanomolar range. This compound also enhances memory in young and very old (30 mo) mice, improves cognitive function and synaptic structure in old (23 mo) AD transgenic mice, and extends the life span of flies. SinceJ147 prevents cell death in multiple cell culture models that lack any of the recognized AD drug targets of the amyloid pathway, it must be acting on highly relevant molecular target(s) and target pathways that are unrelated to the amyloid pathway and that contribute to its therapeutic efficacy. Given that J147 is neuroprotective in multiple, functionally distinct, age-associated toxicity assays, it should be possible to define common, shared neuroprotective pathway(s) which will then be used as the basis for identifying new therapeutic targets for the treatment of AD. This proposed chemical biology approach to identify drug targets originated in 1785 with the isolation of digoxin from foxglove, followed by morphine from poppies in 1806, and aspirin from salicylic acid in 1897. Not only did these molecules make tremendous contributions to medicine, they also were the key factors in the ++ identification of the molecular pathways and drug targets involving the Na -K -ATP pump, opiate receptors, and the cyclooxygenase enzymes, respectively, leading to a succession of derivative drugs that are in the clinic today. The overall goal of this application i to use a chemical biology approach based upon the J147 compound to identify and validate novel, old-age-associated targets for AD treatment and prevention. This will be done by using molecular biology and proteomics to define the shared molecular signaling pathways that lead to neuroprotection in multiple toxicity assays, and chemistry and proteomics to isolate the molecular target(s) of J147. In vivo target validation will be done in part in an animal model that is not biased toward the familial AD/amyloid pathway, but rather reflects most aspects of human aging and AD, the senescence-accelerated SAMP8 mice.