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.

项目摘要/摘要 20％的美国人死于癌症。这种令人震惊的统计数据反映了人类对新颖的主要健康需求 癌症治疗。循环GMP-AMP（CGAMP）的小分子模仿代表新兴的新类别 在最后一次，已将免疫治疗性癌症药物和许多CGAMP仿真药纳入临床试验 三年。不幸的是，由于它们异常复杂的结构，这些临床候选者异常是 很难通过经典的合成方法来准备，这是其发展的限制。最近的 然而，生物催化的进展表明，有很大的机会简化了CGAMP仿真的合成。 自然产生CGAMP的酶称为CGAMP合酶（CGA），初步结果表明 CGA的工程可以通过高度直接的流程提供一种访问潜水员CGAMP仿真的方法。 该提案旨在开发生物催化方法，用于使用自然和 设计的CGA变体。由于已将超过500个CGA酶序列与自然分类，因此 已经是一个大型蛋白质文库，从中开发出扩展的生物催化词典来合成 CGAMP模拟。在合成不自然的CGAMP仿真中面临三个主要挑战，这将是 由这个团契打扮，目的是1）用非本地核碱酶耐受的CGA酶构建一组 Ance，2）为磷光体中心创建立体配置催化剂，3）工程师CGAS创建 CGAMP的非典型宪法异构体。我们的假设是可以实现新颖的反应性和选择性 通过CGAS蛋白的两相迭代询问：CGA库的高通量筛选 针对参考目标的酶，然后是最成功的变体的工程。洞察力 工作将增加对CGA的特定结构特征的了解 Erance，以磷为中心的立体选择性和大环化的位点选择性。这项工作将使Sci- 愿意更快地开发明天的免疫治疗癌药物。