Rapid Selection and Evaluation of Cyclic Peptides in Parkinson's Disease Models

描述（由申请人提供）：我们的实验室在酵母中开发了人类疾病模型，发现了遗传性和散发性蛋白质错误折叠疾病背后的特定细胞缺陷。我们希望利用这些模型来鉴定具有治疗潜力的分子。环肽是一类已被证实的细胞效应，但作为分子治疗的来源，它在很大程度上尚未被探索。最近，我们开发了一种新的高通量方法来筛选小的，遗传编码的环肽分子文库，以防止我们酵母模型中的细胞死亡。环状肽的遗传编码允许以更低的成本更快地筛选数量级更多的化合物，并且比现有程序更少地需要机械化。我们已经将这种方法应用于我们的帕金森病模型，其中过度表达的人类α -突触核蛋白聚集并在酵母中引起毒性。从八聚体环肽库中得到了两个有希望的结果。在这项提议中，我们最初寻求通过构建和筛选几种不同环大小的环肽库来确定更多的命中点。我们将通过检查它们对α -突触核蛋白聚集和细胞器结构的影响，并通过确定它们的蛋白质靶标，来描述撞击的分子机制。我们还将在帕金森病的蠕虫和大鼠模型中测试每一类hit的代表，并开始通过SAR分析优化它们的活性。通过基因编码环肽使得快速选择过程成为可能，并且环肽支架的固有优势使其成为鉴定新化合物的有力方法，这些化合物可以导致蛋白质错误折叠疾病的治疗方法。帕金森氏症是一种毁灭性的神经退行性疾病，影响着超过2%的65岁以上的美国人，因此迫切需要加快寻找治疗帕金森氏症的药物。我们已经在酵母中重现了大部分疾病，酵母是一种非常小的生物，它生长得更快，比现在使用的任何模型系统都更容易操纵。这些显著的优势将使我们能够比现在更快地搜索更多潜在的药物，同时也能检测到迄今为止很少受到关注的有潜力的药物类型。

DESCRIPTION (provided by applicant): Our laboratory has developed models of human disease in yeast that have uncovered specific cellular defects underlying hereditary and sporadic protein misfolding diseases. We wish to make use of these models for the identification of molecules with therapeutic potential. Cyclic peptides are a proven class of cellular effector that is largely unexplored as a source of molecular therapeutics. Recently, we have developed a novel high throughput method of screening libraries of small, genetically encoded cyclic peptides for molecules that prevent cell death in our yeast models. The genetic encoding of the cyclic peptides allows orders of magnitude more compounds to be screened faster, at lower cost, and with less need for mechanization than with existing procedures. We have applied this method to our model of Parkinson's disease, in which overexpressed human alpha-synuclein aggregates and causes toxicity in yeast. Two promising hits from a library of octamer cyclic peptides have resulted. In this proposal, we seek initially to identify more hits by constructing and screening libraries of cyclic peptides of several different ring sizes. We will characterize the molecular mechanisms of the hits that result by examining their effects on alpha-synuclein aggregation and the structure of cellular organelles, and by determining their protein targets. We will also test representatives of each class of hit in worm and rat models of Parkinson's disease, and begin to optimize their activity through SAR analysis. The rapid selection procedure made possible by genetically encoding cyclic peptides, and the inherent advantages of the cyclic peptide scaffold make this a powerful method for identifying new compounds that can lead to therapeutics for protein misfolding diseases. Parkinson's Disease is a devastating neurodegenerative disorder that affects more than 2% of Americans over the age of 65, making the need acute to accelerate ways of finding drugs to treat it. We have recreated much of the disease in yeast, a very small organism that grows much faster and is much more manipulable than any of the model systems now in use. These significant advantages will allow us to search through many more potential drugs and do it much faster than is now possible, as well as to examine types of potentially promising drugs that so far have received little attention.