De novo protein neurotoxins for ion channels

离子通道的从头蛋白神经毒素

基本信息

批准号：
9247713
负责人：
Steve A N Goldstein
金额：
$ 16.47万
依托单位：
BRANDEIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9247713
关键词：
Affect Affinity American Arrhythmia Atrial Fibrillation Bacteriophages Binding Cardiac Cell surface Cells Clinical Cloning Diabetes Mellitus Diagnosis Disease Disulfides Diversity Library Drug Targeting Elan brand of omega-conopeptide MVIIA Fertility Generations Gland Glucose Grant Health Heart Heart Atrium Human Insulin-Dependent Diabetes Mellitus Investigation Ion Channel Ion Channel Protein Laboratories Lead Left Libraries Ligands Membrane Methodology Methods Morbidity - disease rate Neurotoxins Non-Insulin-Dependent Diabetes Mellitus Orphan Peptide Hydrolases Peptides Phage Display Pharmaceutical Preparations Population Potassium Potassium Channel Preparation Proteins Protons Public Health Reporting Resistance Role Scorpions Sorting - Cell Movement Source Specificity Sperm Capacitation Spiders Structure Structure of beta Cell of islet Surface Targeted Toxins Technology Testing Therapeutic Therapeutic immunosuppression Toxin Validation Variant Venoms Voltage-Gated Potassium Channel Work base biophysical techniques combinatorial gene cloning improved inhibitor/antagonist insulin secretion interest male mortality pancreatic juice preclinical study public health relevance receptor scaffold sensor success synthetic peptide targeted treatment therapeutic target tool treatment strategy voltage

项目摘要

DESCRIPTION (provided by applicant): Protein neurotoxins are among the most powerful tools in the arsenal for investigation of membrane receptors and are now used as well for treatment of disease. This grant seeks, first, to advance our method of creation of de novo toxins by targeting potassium and proton channels that lack these useful ligands; second, to use de novo toxins to assess the potential of Kv1.5 and Kv2.1 channels as targets for treatment of two diseases of broad public impact- Atrial Fibrillation (AF) and Type 2 Diabetes (T2D), respectively. Success on these two fronts will allow wider use of these tools to produce toxins for other targets and motivate analogous work directed towards other types of membrane receptors important to human health and disease. To date, toxins have been derived primarily from limited sources: crude venoms, venom gland gene cloning, and laboratory synthesis of known toxins. Therefore, most receptors of interest lack a recognized, specific, high-affinity ligand. This state-of-affairs is easily understood: neither the purpose of toxins in the wild nor screens of venoms favor target specificity. Here, these problems are avoided by cloning toxins based on desired attributes using a new, high- throughput method we developed to generate a selective inhibitor of Kv1.3 channel pore. Specific Aim 1 seeks to improve the method by creating highly-diverse libraries on native scaffolds that bind to ion channel pores or voltage sensors, first using purified ion channel receptors, and then using channels on cell surfaces so the many receptors that do not retain their structure when purified can also be targeted. A recently identified channel for protons (Hv1) is used here to develop tools for voltage sensors (and to test directly the postulated role of this orphan receptor in male fertility). Specific Aim seeks de novo toxins directed to the Kv1.5 potassium channel. This orphan receptor is widely pursued as a potential therapeutic target for AF and lack of specific blockers has left unclear fundamental questions about its role in the heart and precluded direct tests of its utility as a drg target. Specific Aim 3 seeks de novo toxins specific for the Kv2.1 channel. Kv2.1 inhibition enhances glucose-stimulated insulin secretion from pancreatic -cells in a SUMO-dependent manner and inhibitors that do not cross inhibit are sought as potential drugs for T2D. De novo toxins for Kv1.5 and Kv2.1 are not only useful to validate the channels as drug targets but offer potential as therapeutic leads. Thus, small, disulfide-stabilized peptide toxins in use as drugs (Byetta(r), Integrilin(r), and Prialt(r)) have proven effective, protease resistant and non-immunogenic.

描述（由适用提供）：蛋白质神经毒素是用于投资膜受体的最强大工具之一，现在也用于治疗疾病。该赠款首先寻求通过靶向缺乏这些有用的配体的钾和质子渠道来推动我们创建从头毒素的方法。其次，使用从头毒素来评估KV1.5和KV2.1通道的潜力，作为治疗两种广泛的公共影响 - 房颤（AF）和2型糖尿病（T2D）的靶标。在这两个方面的成功将使这些工具更广泛地用于其他靶标生产毒素，并融合针对其他对人类健康和疾病重要的膜受体的类似工作。迄今为止，毒素主要来自有限的来源：粗毒液，毒腺基因克隆和实验室合成已知的毒素。因此，大多数感兴趣的接收者都缺乏公认的，特定的高亲和力配体。这种最新的伴随很容易理解：野生毒素的目的也不是毒素筛选的目的。在这里，通过基于所需属性克隆毒素，使用一种新的高通量方法来避免这些问题，我们开发了用于生成KV1.3通道孔的选择性抑制剂。特定的目标1试图通过在与离子通道孔或电压传感器结合的天然支架上创建高度多样性的文库，首先使用纯化的离子通道受体，然后在细胞表面上使用通道，从而将其当纯化时不保留其结构的许多受体也可以针对目标。这里使用了最近确定的质子通道（HV1）来开发用于电压传感器的工具（并直接测试该孤儿受体在男性生育中的作用）。具体目的是寻求针对KV1.5钾通道的从头毒素。该孤儿接收器被广泛作为AF的潜在治疗靶标，并且缺乏特定的阻滞剂对其在心脏中的作用以及排除其作为DRG目标的实用性的直接测试留下了不清楚的基本问题。特定的目标3寻求对KV2.1渠道特定的从头毒素。 Kv2.1抑制作用以SUMO依赖性方式增强了胰腺细胞的葡萄糖刺激的胰岛素分泌，并且不交叉抑制的抑制剂被视为T2D的潜在药物。 KV1.5和KV2.1的从头毒素不仅有助于验证通道作为药物靶标，而且具有治疗铅的潜力。该用作药物（BYETTA（R），整合蛋白（R）和Prialt（R））的小型，二硫键稳定的肽毒素已被证明有效，抗蛋白酶耐药性和非免疫原性。