Biocatalytic approaches to antiepileptic drug targets

抗癫痫药物靶标的生物催化方法

• 批准号: 9761129
• 负责人: April Lukowski
• 金额: $ 3.66万
• 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 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9761129
• 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; Techniques; Toxin; Unspecified or Sulfate Ion Sulfates; 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性和特异性的小分子配体 疾病相关的同种型具有挑战性。常见的合成策略需要预官能化以引入 杂原子和更大的官能团，使得分子难以处理、纯化和经受 进一步多样化。大自然通过构建简单的核心来解决这种合成瓶颈， 在生物合成后期装饰支架。化学家从大自然的技术中获得灵感， 晚期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的因此，生物催化代表了一种独特的方法来解决 与药物设计相关的合成挑战。 麻痹性贝类毒素是一种尚未开发的抗癫痫药物靶点。超过50种天然来源 已识别出PSTs，并已评估了少数与电压门控钠的结合 已经证明了阻断VGSC通道（VGSC）的能力。这种分子反应对应于 抗癫痫药物靶点所需的物理反应。PSTs作为抗癫痫药物靶点的研究 由于具有挑战性的合成路线和无法分离足够量的大多数>50 类似物与麻痹性贝类毒素生物合成相关的基因簇已经被鉴定， 利用酶的机会，即使是最熟练的化学家也无法获得化学能力。这 该提案描述了阐明麻痹性贝类毒素生物合成途径、评估酶 底物范围，并从生物催化反应中分离新型化合物，用于使用VGSC进行分析， 电生理技术。 总之，这项工作的目的是多样化的PST支架使用PST生物合成酶从蓝藻， 使得能够在复杂的、富含杂原子的分子上进行化学转化，否则这些分子是难以处理的。的 该提案中建立的方法将通过开发新的抗癫痫药物来加速发现新的抗癫痫药物。 使用来自已知VGSC阻断化合物的生物合成途径的生物催化剂的化学反应。