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

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/10534903
• 关键词: Address; Alkaloids; Anabolism; Antimalarials; Binding; Bioinformatics; Biological; Biological Testing; Combinatorial Synthesis; Coupled; Coupling; Cytochrome P450; Detection; Development; Directed Molecular Evolution; Drug Design; Drug Targeting; Engineering; Enzymes; Evaluation; Gene Cluster; Goals; Hydrogen Bonding; Image; Indoles; Investigation; Lead; Libraries; Methods; Minor; Natural Products; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Plants; Preparation; Property; Protein Engineering; Reaction; Research; Rotation; Route; Site; Specificity; Stereoisomer; Structure-Activity Relationship; Testing; Therapeutic; Variant; analog; base; biological systems; chiral molecule; design; drug development; high throughput screening; improved; interest; malaria transmission; monomer; physical property; racemization; scaffold; screening; single bond; small molecule; tool; transmission process

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.

项目总结/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， 传统上被避免作为药物设计中的设计策略。作为这一结果， 尽管越来越多的证据表明，阻转异构体可以是不发达的， 策略性地应用于通过增加其结合来改善潜在药物的效力和选择性 对生物目标的特异性。特别地，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轴的手性聚合物通常依赖于具有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实用性。在这里，我建议 开发了一种生物催化方法用于二芳基醚的合成，C，N-偶联 萘异喹啉生物碱和N-芳基吲哚。这些类别的化合物是基于 目前的限制，他们的合成和他们的潜在的治疗性质，与拟议的研究 旨在解决这两点。为达致这个目标，我们会分三个阶段进行，包括 筛选野生型酶、生物催化剂工程、组合合成和生物测试 复合图书馆。 在第一阶段，将针对野生型P450酶文库筛选单体。此库 使用生物信息学方法构建，以鉴定与那些具有序列相似性的P450酶。 已知的反应性。在这个阶段，将分析反应的反应性证据和检测的 目标产品。一旦确定了合适的工程起点，就可以指导 酶将被引导以改善目标产物的形成。最后，一旦找到合适的P450变体， 鉴定后，将在96孔板中通过组合合成构建化合物文库。这些 然后将对化合物进行生物学测试，通过分析它们的抗疟潜力， 影响传播和生存能力。上述研究建议将有助于高彻底性 实验和反应分析，结合生物测试的高通量平台。 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 访问。这将加速潜在治疗化合物的制备和评估，加快药物的开发。 确定抗疟药物的目标。