Biocatalytic Enantioselective Synthesis of Non-Biaryl and Hetero-Biaryl Atropisomers and Testing of their Antimalarial Properties

项目摘要/摘要 异构化，或由于键旋转受阻而产生的立体异构化， 传统上在药物设计中避免将其作为一种设计策略。因此，催化方法可用于 阿托品选择性合成不发达，尽管越来越多的证据表明阿托品异构症 战略性地应用于通过增加潜在药物的结合力来提高其效力和选择性 对生物靶标的专一性。特别是非联芳基和杂联芳基异构体的合成 含有手性C-O和C-N轴通常依赖于具有C-X键的底物的官能化 已经安装，从而限制了这些方法在设计收敛合成中的应用。在这里，我建议 生物催化合成C，N偶联二芳醚的研究进展 萘基异喹啉生物碱和N-芳基吲哚。这些化合物的类别是根据 目前对它们的合成和潜在治疗特性的限制，以及拟议的研究 旨在解决这两个问题。为了达到这一目标，我们将采取三个阶段的方法，包括 野生型酶的筛选，生物催化剂工程，组合合成和生物测试 复合库。 在第一阶段，将根据野生型P450酶的文库筛选单体。这个图书馆 使用生物信息学方法构建，以鉴定P450酶，其序列与 具有已知的反应性。在这个阶段，将对反应进行分析，以寻找反应性的证据和检测 目标产品。在确定了合适的工程起点后，定向进化 将进行酶促目标产物的形成。最后，一旦一个合适的P450变体 经过鉴定，将在96个井板上通过组合合成构建化合物文库。这些 然后，化合物将接受生物测试，以评估其抗疟疾潜力，通过分析它们的 对传播和生存能力的影响。以上提出的研究将以高通量为基础 实验和反应分析，配合高通量生物检测平台。 我们预计，这项研究将简化以前具有挑战性的分子类的合成 才能访问。这将加快潜在治疗化合物的制备和评估，加快 确定抗疟疾药物靶标。

Project Summary/Abstract Atropisomerism, or stereoisomerism arising as a consequence of hindered bond rotation, has traditionally been avoided as a design strategy in drug design. As a result of this, catalytic methods for atroposelective synthesis are underdeveloped, despite the growing evidence that atropisomerism can be strategically applied to improve the potency and selectivity of a potential drug by increasing its binding specificity to a biological target. In particular, the synthesis of non-biaryl and hetero-biaryl atropisomers containing chiral C–O and C–N axes often relies on the functionalization of a substrate with the C–X bond already installed, thus limiting the utility of these methods in designing convergent syntheses. Here, I propose the development of a biocatalytic approach for the atroposelective synthesis of diarylethers, C,N-coupled naphthylisoquinoline alkaloids, and N-aryl indoles. These classes of compounds were chosen based on the current limitations for their syntheses and their potential therapeutic properties, with the proposed research aiming to address both points. To achieve this goal, we will take a three stage approach consisting of screening wild-type enzymes, biocatalyst engineering, and combinatorial synthesis and biological testing of compound libraries. In the first stage, monomers will be screened against a library of wild-type P450 enzymes. This library was constructed using a bioinformatics approach to identify P450 enzymes with sequence similarity to those with known reactivity. In this stage, reactions will be analyzed for evidence of reactivity and detection of the target product. Upon identification of a suitable starting point for engineering, directed evolution of the enzyme will be conducted to improve formation of the target product. Finally, once a suitable P450 variant is identified, a library of compounds will be constructed through combinatorial synthesis in 96 well plates. These compounds will then undergo biological testing to evaluate their antimalarial potential by analyzing their impact on transmission and viability. The research proposed above will be facilitated by high-thoroughput experimentation and reaction analysis, in conjunction with high-thoroughput platform for biological testing. We anticipate that that this research will streamline the synthesis of molecule classes previously challenging to access. This will accelerate preparation and evaluation of potential therapeutic compounds, expediting the identification of antimalarial drug targets.