Biocatalytic approaches to antiepileptic drug targets

建议书摘要 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 与疾病相关的亚型是具有挑战性的。常见的综合策略需要预先功能化才能引入 杂原子和更大的官能团，使分子难以处理、提纯和受制于 进一步多元化。大自然通过构建简单的核心和 在以后的生物合成中装饰支架。化学家从大自然的技术中获得设计灵感 后期的C-H功能化途径，但酶产生分子复杂性的能力是 是最先进的合成方法所无法比拟的。因此，生物催化是一种独特的解决方法。 与药物设计相关的合成挑战。 麻风性贝类毒素(PST)是抗癫痫药物靶标的未开发来源。超过50个天然来源 已经确定了PSTs，并选择了少数几个被评估为与电压门控钠结合的PSTs 通道(VGSC)已经展示了阻止VGSC的能力。这种分子反应对应于 抗癫痫药物靶点所需的生理反应。PSTs作为抗癫痫药物靶点的研究进展 由于具有挑战性的合成路线以及无法分离出足够数量的大多数&gt；50 类比。与麻痹性贝类毒素生物合成相关的基因簇已经被发现，使 即使是最熟练的化学家也无法获得利用能够产生化学作用的酶的机会。这 方案描述了阐明麻痹性贝类毒素生物合成途径、评价酶的策略 底物范围，并从生物催化反应中分离出新化合物，用于VGSCS分析 电生理技术。 综上所述，本工作旨在利用蓝藻合成PST的酶使PST支架多样化， 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。这个 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封闭化合物的生物合成途径的生物催化剂的化学反应。

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.