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。这一令人震惊的统计数字反映了人类对小说的主要健康需求 癌症治疗学。环状GMP-AMP的小分子模拟物(CGAMP)代表了一类新兴的 免疫治疗癌症药物和许多cGAMP-模拟物在过去的几年中已经进入临床试验 三年了。不幸的是，由于它们异常复杂的结构，这些临床候选对象是例外的 很难通过经典的合成方法进行准备，这对它们的发展构成了限制。近期 然而，生物催化方面的进展表明，简化cGAMP-模拟物的合成具有重要的机会。 自然产生cGAMP的酶被称为cGAMP合成酶(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得各种cGAMP-模拟物的方法。 这项建议旨在开发生物催化方法，用于合成cGAMP-模拟物，使用天然和 经过改造的cGAS变种。由于已从自然界中编目了500多个cGAS酶序列， 已经是一个很大的蛋白质文库，可以从中开发一个扩展的生物催化词典来合成 CGAMP-模拟。在非天然cGAMP-模拟物的合成中存在三个主要挑战，这些挑战将被认为是 该奖学金旨在1)用非天然的碱基Toler构建一组cGAS酶-- 2)为硫代中心创造立体互补催化剂，3)工程cGAS以创造 CGAMP的非典型构型异构体。我们的假设是可以实现新的反应性和选择性 通过两阶段重复询问cGAS蛋白：高通量筛选cGAS文库 酶对照参考目标，然后设计出最成功的变种。从这一点上的洞察 这项工作将增加对cGAS特定结构特征的了解，这些结构特征支配着碱基TOL- 大环化反应中的磷中心立体选择性和位置选择性。这项工作将使SCI- 以更快的速度开发未来的免疫治疗癌症药物。