Novel Pharmacological Agents from Conus regius

描述(由申请人提供)：这一领域的提案旨在从西大西洋锥形蜗牛种--注册圆锥螺的毒液中分离和表征新的成分。锥形蜗牛是一种海洋软体动物，用毒液捕捉猎物。蛇毒的组成成分可引起广泛的神经生理反应，并显示出巨大的治疗潜力。在这项提议中，我们将研究新的王冠扇贝蛇毒中的鱼腥多肽成分。注册圆锥具有独特的生物地理学特性，我们的初步数据表明，它具有广泛的新成分，具有明显的神经调节特性。我们建议利用我们实验室开发的高分辨率层析方案，有效地将复杂的华支睾吸虫毒液分离成单一组分。这些由本地毒液提供的纳摩尔量分离的单一成分，将通过我们实验室开发的新的生物分子核磁共振技术进行分析。这些非破坏性技术依赖于使用在低温下工作的高分辨率/魔角旋转探头，以及使用脉冲场梯度和选择性激发的同核和异核2D-核磁共振实验。核磁共振数据将得到使用fempto-摩尔水平的ESI-MS/MS技术的补充，这将使我们能够以非破坏性的方式对翻译后大量修饰的椰子多肽进行核磁共振/MS测序。这些分析将与使用基于荧光的/特定于金属离子的小鼠皮质细胞的高通量体外功能生物检测技术一起完成。这些分析将评估从王冠花中提取的蛇床子多肽的神经调节特性。我们将产生一组新的刀豆肽，除了它们潜在的治疗价值外，它们还可以用作分子探针，从而能够确定神经元受体的功能和作用机制。

DESCRIPTION (provided by applicant): This AREA proposal is aimed at the isolation and characterization of novel components from the venom of Conus regius, a Western Atlantic cone snail species. Cone snails are marine mollusks that use venom to capture their prey. The components of the venom (conopeptides) elicit a wide range of neurophysiological responses and show great potential as therapeutic agents. In this proposal we will investigate the novel conopeptides components of the venom of C. regius. Conus regius is biogeographically unique and our preliminary data shows that it possesses a wide array of new components with distinct neuromodulatory characteristics. We are proposing to utilize a high-resolution chromatographic scheme, developed in our laboratory, to efficiently separate the complex venom of C. regius into single components. These single components, isolated in nanomolar quantities provided by the native venom, will be analyzed by novel biomolecular NMR techniques, also developed in our lab. These nondestructive techniques rely on the use of a high resolution/magic angle spinning probe operating at low temperatures with homonuclear and heteronuclear 2D-NMR experiments that use pulse field gradients and selective excitation. The NMR data will be complemented by the use fempto-molar level ESI-MS/MS techniques that will allow us to carry out NMR/MS sequencing of the heavily post-translationally modified conopeptides in a non-destructive fashion. These analyses will be done in conjunction with high throughput in vitro functional bioassays using fluorescent-based/metal ion-specific techniques on murine cortical cells. These assays will assess the neuromodulatory characteristics of the conopeptides from C. regius. We will generate a novel set of conopeptides, which in addition of their potential therapeutic value; they can be used as molecular probes that would allow for the determination of function and mechanisms of actions of neuronal receptors.