Biocatalytic approaches to antiepileptic drug targets

抗癫痫药物靶标的生物催化方法

• 批准号: 9922670
• 负责人: April Lukowski
• 金额: $ 1.2万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9922670
• 关键词: Action Potentials; Affinity; Anabolism; Antiepileptic Agents; Architecture; Arrhythmia; Binding; Biochemical; Biological Assay; Brain; Breathing; Cardiac; Chemicals; Chemistry; Childhood; Climacteric; Complex; Cyanobacterium; Development; Disease; Dose; Drug Design; Drug Targeting; Electrophysiology (science); Enzymes; Epilepsy; Family; Gene Cluster; Generations; Genes; Guanidines; Human; Human body; Hydroxylation; Individual; Lead; Ligands; Mediating; Methods; Molecular; Mus; Mutate; Mutation; Myotonia; Natural Products; Nature; Neuraxis; Neurons; Paralysed; Pathway interactions; Patients; Pattern; Positioning Attribute; Protein Isoforms; Rattus; Reaction; Reporting; Research; Route; Sampling; Saxitoxin; Shellfish; Signal Transduction; Skeletal Muscle; Sodium Channel; Sodium Channel Binding; Sodium Channel Blockers; Source; Specificity; Sulfate; Techniques; Toxin; Variant; Work; analog; base; chemical reaction; design; dravet syndrome; drug market; enzyme substrate; epileptic encephalopathies; functional group; gonyautoxins; hydroxyl group; novel; oxidation; response; scaffold; side effect; skeletal; small molecule; student mentoring; sulfotransferase; voltage

Proposal Summary Discovering small molecule ligands with a high affinity for voltage-gated sodium channels and specificity for disease relevant isoforms is challenging. Common synthetic strategies require prefunctionalization to introduce heteroatoms and larger functional groups, rendering the molecules difficult to handle, purify, and subject to further diversification. Nature approaches such synthetic bottlenecks by constructing simple cores and decorating scaffolds later on in biosynthesis. Chemists take inspiration from Nature’s techniques in designing late-stage C–H functionalization routes, but the ability of enzymes to generate molecular complexity is unmatched by state-of-the-art synthetic methods. Thus, biocatalysis represents a unique approach to tackling the synthetic challenges associated with drug design. Paralytic shellfish toxins (PSTs) are an untapped source of antiepileptic drug targets. Over 50 naturally derived PSTs have been identified, and the select few that have been assessed for binding to voltage-gated sodium channels (VGSCs) have demonstrated the ability to block VGSCs. This molecular response corresponds to physical responses desired in antiepileptic drug targets. The study of PSTs as antiepileptic drug targets has been hindered by challenging synthetic routes and the inability to isolate sufficient quantities of most of the >50 analogs. Gene clusters associated with paralytic shellfish toxin biosynthesis have been identified, enabling opportunities to leverage enzymes capable of chemistry inaccessible to even the most skilled chemist. This proposal describes strategies to elucidate the paralytic shellfish toxin biosynthetic pathway, evaluate enzyme substrate scopes, and isolate novel compounds from biocatalytic reactions for analysis with VGSCs using electrophysiological techniques. In summary, this work aims to diversify the PST scaffold using PST biosynthetic enzymes from cyanobacteria, enabling chemical transformations on complex, heteroatom-rich molecules that are otherwise intractable. The methods established in this proposal will accelerate the discovery of new antiepileptic drugs by developing new chemical reactions using biocatalysts from the biosynthetic pathway of known VGSC blocking compounds.

提案摘要 发现对电压门控钠通道具有高亲和力和特异性的小分子配体 疾病相关亚型具有挑战性。常见的合成策略需要预功能化来引入 杂原子和较大的官能团，使得分子难以处理、纯化和经受 进一步多元化。大自然通过构建简单的核心来解决此类合成瓶颈 随后在生物合成中装饰支架。化学家从大自然的设计技术中汲取灵感 后期 C-H 功能化路线，但酶产生分子复杂性的能力是 最先进的合成方法无法比拟。因此，生物催化代表了解决这一问题的独特方法。 与药物设计相关的综合挑战。 麻痹性贝类毒素（PST）是抗癫痫药物靶标的尚未开发的来源。超过 50 种自然衍生的 PST 已被鉴定，并且选定的少数几个已被评估与电压门控钠的结合 通道（VGSC）已证明具有阻止 VGSC 的能力。该分子反应对应于 抗癫痫药物靶标所需的物理反应。 PST 作为抗癫痫药物靶点的研究 由于具有挑战性的合成路线以及无法分离足够数量的大多数> 50 类似物。与麻痹性贝类毒素生物合成相关的基因簇已被鉴定，使得 即使是最熟练的化学家也无法利用具有化学能力的酶的机会。这 提案描述了阐明麻痹性贝类毒素生物合成途径、评估酶的策略 底物范围，并从生物催化反应中分离出新化合物，以便使用 VGSC 进行分析 电生理学技术。 总之，这项工作旨在利用蓝藻中的 PST 生物合成酶使 PST 支架多样化， 能够对复杂的、富含杂原子的分子进行化学转化，否则这些分子很难处理。这 该提案中建立的方法将通过开发新的抗癫痫药物来加速新的抗癫痫药物的发现 使用来自已知 VGSC 阻断化合物生物合成途径的生物催化剂进行化学反应。