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.

项目摘要/摘要因阻碍粘结旋转而引起的逆性异构体或立体异构主义的具有传统上，避免了作为药物设计的设计策略。因此，催化方法阳性性合成不发达，dospite越来越多的证据表明肉骨分散性可以是从策略上应用来通过增加其结合来提高潜在药物的效力和选择性生物靶标的特异性。特别是，非生物和杂脂异构体的合成含有手性C – O和C – N轴通常依赖于C – X键的底物的功能化已经安装了，因此限制了这些方法在设计收敛合成中的实用性。我建议在这里开发日记室的肿瘤性合成的生物催化方法C，N偶联萘基喹啉生物碱和N-芳基吲哚。这些化合物是根据拟议的研究旨在解决这两个要点。为了实现这一目标，我们将采用三个阶段的方法筛选野生型酶，生物催化剂工程以及组合合成和生物学测试复合库。在第一阶段，将对野生型P450酶的库进行筛选。这个库是使用生物信息学方法构建的，以识别与序列相似性的P450酶具有已知的反应性。在此阶段，将分析反应的反应性证据和检测目标产品。在确定了适当的工程起点时，将进行酶以改善目标产物的形成。最后，一旦合适的P450变体是鉴定出，化合物库将通过96个井板中的组合合成来构建。这些然后，化合物将经过生物学测试，以通过分析它们对传输和生存能力的影响。上面提出的研究将由高途径准备实验和反应分析与用于生物学测试的高震动平台结合使用。我们预计这项研究将简化以前挑战的分子类的合成访问。这将加快潜在治疗化合物的准备和评估，并加快鉴定抗疟药靶标。