Development of a Biocatalytic Toolbox for the Synthesis of Small-Molecule Mimics of cyclic GMPAMP

开发用于合成环状 GMPAMP 小分子模拟物的生物催化工具箱

• 批准号: 10751277
• 负责人: Cole Christian Meyer
• 金额: $ 6.87万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751277
• 关键词: Address; American; Antineoplastic Agents; Chemicals; Clinical; Clinical Trials; Communities; Complex; Constitution; Constitutional; Cyclic GMP; Data; Development; Dinucleoside Phosphates; Drug Compounding; Drug Targeting; Eating; Engineering; Enzymes; Fellowship; Friends; Generations; Health; Homologous Gene; Human; Hydroxyl Radical; Immunotherapeutic agent; Investigation; Isomerism; Knowledge; Libraries; Malignant Neoplasms; Methodology; Methods; Modification; Nature; Nucleosides; Periodicity; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phosphorus; Preparation; Process; Protein Engineering; Proteins; Reference Standards; Reporting; Research; Route; STING agonists; Scientist; Site; Structure; Technology; Therapeutic; Variant; Vertebral column; Work; analog; cancer immunotherapeutics; cancer therapy; catalyst; clinical candidate; cost; drug efficacy; high throughput screening; improved; inorganic phosphate; insight; manufacture; non-Native; novel; novel therapeutics; nucleobase; phosphorothioate; screening; small molecule; statistics

PROJECT SUMMARY/ABSTRACT Twenty percent of Americans die of cancer. This alarming statistic reflects a major human health need for novel cancer therapeutics. Small molecule mimics of cyclic GMP-AMP (cGAMP) represent an emerging new class of immunotherapeutic cancer drugs, and numerous cGAMP-mimics have been entered into clinical trials in the last three years. Unfortunately, due to their unusually complex structure, these clinical candidates are exceptionally difficult to prepare via classic synthetic methodology, which serves as a limitation to their development. Recent advances in biocatalysis, however, indicate significant opportunity to simplify the synthesis of cGAMP-mimics. The enzyme that naturally produces cGAMP is called cGAMP synthase (cGAS), and preliminary results suggest that engineering of cGAS may provide a way to access diverse cGAMP-mimics through a highly direct process. This proposal aims to develop biocatalytic approaches for the synthesis of cGAMP-mimics using natural and engineered cGAS variants. As over 500 sequences of cGAS enzymes have been catalogued from nature, there is already a large protein library from which to develop an expanded biocatalytic lexicon for the synthesis of cGAMP-mimics. There are three major challenges in the synthesis of unnatural cGAMP-mimics that will be ad- dressed by this fellowship, which aims to 1) build a panel of cGAS enzymes with non-native nucleobase toler- ance, 2) create stereocomplementary catalysts for phosphorothioate centers, and 3) engineer cGAS to create atypical constitutional isomers of cGAMP. It is our hypothesis that novel reactivity and selectivity can be realized through a two-phase iterative interrogation of cGAS proteins: high-throughput screening of a library of cGAS enzymes against a reference target followed by engineering of the most successful variants. Insights from this work will afford an increased knowledge of the specific structural features of cGAS that govern nucleobase tol- erance, phosphorus-centered stereoselectivity, and site-selectivity in macrocyclization. This work will enable sci- entists to develop tomorrow’s immunotherapeutic cancer drugs more rapidly.

项目概要/摘要 百分之二十的美国人死于癌症。这一令人震惊的统计数据反映了人类健康对新型药物的主要需求 癌症治疗。环 GMP-AMP (cGAMP) 的小分子模拟物代表了一类新兴的新型药物 近年来，免疫治疗癌症药物和众多cGAMP模拟物已进入临床试验 三年。不幸的是，由于其异常复杂的结构，这些临床候选者异常 通过经典的合成方法很难制备，这限制了它们的发展。最近的 然而，生物催化的进步表明有机会简化 cGAMP 模拟物的合成。 天然产生 cGAMP 的酶称为 cGAMP 合酶 (cGAS)，初步结果表明 cGAS 的工程可能提供一种通过高度直接的过程获取多种 cGAMP 模拟物的方法。 该提案旨在开发利用天然和合成 cGAMP 模拟物的生物催化方法。 工程化的 cGAS 变体。由于自然界中已经对 500 多个 cGAS 酶序列进行了编目，因此 已经是一个大型蛋白质库，可以从中开发扩展的生物催化词典来合成 cGAMP-模拟物。非天然 cGAMP 模拟物的合成存在三个主要挑战： 该奖学金的目的是 1) 建立一组具有非天然核碱基耐受性的 cGAS 酶 - ance，2) 为硫代磷酸酯中心创建立体互补催化剂，以及 3) 设计 cGAS 以创建 cGAMP 的非典型结构异构体。我们的假设是可以实现新颖的反应性和选择性 通过 cGAS 蛋白的两阶段迭代询问：cGAS 文库的高通量筛选 针对参考目标的酶，然后设计最成功的变体。由此得出的见解 工作将提供更多关于控制核碱基 tol 的 cGAS 特定结构特征的知识 erance、以磷为中心的立体选择性和大环化中的位点选择性。这项工作将使科学 科学家们更快地开发未来的免疫治疗癌症药物。