Novel Neuronal nAChR-Targeted Peptides

描述(申请人提供)：选择性操作烟碱型乙酰胆碱受体(NAChRs)的功能可能有益于治疗各种影响精神和神经健康的疾病，包括情绪障碍、帕金森病、认知障碍和慢性疼痛。目前研究的一个主要方向是确定nAChRs不同功能通道亚型中的亚基，并了解它们在健康和疾病中的作用。针对nAChR亚型的选择性探针的可获得性将极大地促进这些努力，并为开发能够调节神经系统中特定nAChR中介功能的药物提供平台。圆锥螺属肉食性海洋蜗牛的毒液是一种非常丰富的化合物(α-螺毒素)，可以有效和选择性地针对特定的nAChR亚型。这项建议的目的是鉴定和表征这些毒液中区别于神经元nAChRs不同的乙酰胆碱结合亚单位接口的化合物。这一目标将通过使用基于受体的分析来跟踪活性毒液化合物的纯化来实现。还将克隆编码α-芋螺毒素的基因，以分离新的毒素。无论采用哪种方法鉴定的多肽都将进行生化表征和合成。然后，将根据受体亚型特异性对这些多肽进行充分的表征。随后，将利用合成策略来开发具有精细特异性的第二代配体。结构/功能研究将确定关键的受体结合残基。使用多种方法的组合，包括使用选择性的芋螺毒素探针，将从功能和生物化学上确定天然受体的分子组成。

DESCRIPTION (provided by applicant): Selective manipulation of the function of nicotinic acetylcholine receptors (nAChRs) may be beneficial in the treatment of a variety of disorders that impact mental and neurological health, including mood disorders, Parkinson's Disease, cognitive disorders and chronic pain. A major thrust of current research is to identify the subunits in the different functional channel subtypes of nAChRs and to understand their roles in health and disease. The availability of selective probes for the nAChR subtypes will greatly facilitate these efforts, and provide a platform for development of medications capable of modulating specific nAChR-mediated functions in the nervous system. Venoms from carnivorous marine snails of the genus Conus are an extraordinarily rich source of compounds (alpha-conotoxins) that potently and selectively target specific nAChR subtypes. The aim of this proposal is to identify and characterize compounds from these venoms that discriminate among distinct acetylcholine-binding subunit interfaces of neuronal nAChRs. This aim will be accomplished by using receptor-based assays to track the purification of active venom compounds. Genes encoding alpha-conotoxins will also be cloned to isolate new toxins. Peptides identified by either approach will be biochemically characterized and synthesized. These peptides will then be fully characterized with respect to receptor subtype specificity. Subsequently, synthetic strategies will be utilized to develop second-generation ligands with refined specificity. Structure/function studies will determine critical receptor binding residues. Using a combination of approaches, including the use of selective conotoxin probes, the molecular composition of native receptors will be functionally and biochemically defined.