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.

项目总结/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药物的主要需求。 癌症治疗学环GMP-AMP（cGAMP）的小分子模拟物代表了一类新兴的新的生物活性物质。 免疫抗癌药物和许多cGAMP模拟物已进入临床试验，在过去的几年里， 三年不幸的是，由于它们异常复杂的结构，这些临床候选物异常地 难以通过经典的合成方法制备，这限制了它们的发展。最近 然而，生物催化的进展表明简化cGAMP模拟物的合成的重要机会。 天然产生cGAMP的酶被称为cGAMP合成酶（cGAS），初步结果表明， cGAS的工程化可以提供一种通过高度直接的过程来获得不同的cGAMP模拟物的方法。 该提案旨在开发使用天然和天然的生物催化剂合成cGAMP模拟物的生物催化方法。 工程化cGAS变体。由于已经从自然界中编目了超过500种cGAS酶的序列， 已经是一个大的蛋白质库，从中可以开发一个扩展的生物催化词典，用于合成 cGAMP模拟物。在合成非天然cGAMP模拟物中存在三个主要挑战， 由该奖学金装扮，其目的是1）建立一组具有非天然核碱基耐受的cGAS酶， 例如，2）创建用于硫代磷酸酯中心的立体互补催化剂，以及3）工程化cGAS以创建 cGAMP的非典型结构异构体。这是我们的假设，新的反应性和选择性可以实现 通过cGAS蛋白的两阶段迭代询问：cGAS文库的高通量筛选 针对参考靶标的酶，然后工程化最成功的变体。从中获得的见解 工作将提供更多的知识，具体的结构特征的cGAS，支配核碱基tol- 在大环化反应中，磷中心的立体选择性和位点选择性。这项工作将使科学- 更快地开发明天的免疫抗癌药物。