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

非联芳基和杂联芳基阻转异构体的生物催化对映选择性合成及其抗疟性能测试

基本信息

批准号：
10534903
负责人：
Casey Bard Roos
金额：
$ 6.68万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-15 至 2025-08-14
项目状态：
未结题

项目摘要

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.

项目概要/摘要阻转异构现象，或由于键旋转受阻而产生的立体异构现象，传统上在药物设计中避免使用催化方法。尽管越来越多的证据表明阻转异构现象可以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特别是非联芳基和杂联芳基阻转异构体的合成。含有手性 C-O 和 C-N 轴通常依赖于具有 C-X 键的底物的功能化已经安装，因此限制了这些方法在设计收敛合成中的效用。开发用于 C,N 偶联二芳基醚的生物催化方法萘基异喹啉生物碱和 N-芳基吲哚这些类别的化合物是根据以下性质选择的。目前其合成的局限性及其潜在的治疗特性，以及拟议的研究为了解决这两点，我们将采取三阶段方法：野生型酶的筛选、生物催化剂工程以及组合合成和生物测试化合物库。在第一阶段，将针对野生型 P450 酶库筛选单体。使用生物信息学方法构建，以鉴定与那些具有相似序列的 P450 酶在此阶段，将分析反应性的证据并检测反应性。确定合适的工程起点后，即可定向开发目标产品。最后，一旦找到合适的 P450 变体，将进行酶促反应以改善目标产物的形成。确定后，将通过在 96 孔板中组合合成来构建化合物库。然后，化合物将接受生物测试，通过分析其抗疟潜力来评估其抗疟潜力。高通量将促进上述研究。实验和反应分析，结合高通量生物测试平台。我们预计这项研究将简化以前具有挑战性的分子类别的合成这将加速潜在治疗化合物的制备和评估，从而加快开发速度。抗疟药物靶点的鉴定。