Tools to Study Electrophilic Stress and Develop Covalent Drugs

摘要 活性电泳体物种不断地修饰生物分子并以至今仍在影响人类健康的方式 人们对此知之甚少。本研究方案为严谨检验奠定了方法学基础 关于活性亲电物种的假说和开发新的共价不可逆药物。生物活性 亲电性物质，如α，β-不饱和羰基化合物，存在于环境和饮食中，但它们也 在新陈代谢和氧化应激过程中内源性产生的。它们被认为与多种疾病的病因有关。 病理状态，如动脉粥样硬化、癌症、神经变性和炎症。了解 活性亲电物种在动态平衡和疾病中的作用将为疾病提供重要的指导 预防和开发新的治疗方法。然而，缺乏研究工具阻碍了这一点 这一领域的努力和许多研究缺乏实验的严谨性。问题是这些反应性的亲电性 物种不可逆地改变了大量的生物分子，科学家几乎不能控制。该计划 将填补亲电体在生物学效应研究中的关键方法学空白。预期的 可交付成果是能够在特定时间控制此类亲电体生成的方法，以及 地点。此外，可控制地逆转形成的这种亲电性的共价加合物的方法将 被开发出来。此外，筛选技术以开发针对特定蛋白质残留物的分子将 被建立起来。这些方法是创新的，因为它们允许回答重要的生物医学问题 目前还遥不可及。该计划的初步研究已经为实现 这些目标。我们已经开发了化学反应，可以按需生成α，β-不饱和羰基 在生命系统中，我们已经建立了DNA编码库，用于发现生物活性物种。使用 有了这些独特的能力，研究计划就有能力推进对 生物学中的活性亲电物种及其对疾病的影响。为了演示它们的效用，我们将应用 以研究4-羟基壬烯醛在动脉粥样硬化中的作用，并研究适应状态的动力学。 与胰腺癌KRASG12C抑制剂的耐药性有关。我们会采取措施，确保 开发的方法将可供广泛的科学家使用，以最大限度地发挥该计划的影响。

Abstract Reactive electrophile species constantly modify biomolecules and impact human health in ways that are still poorly understood. This research program establishes the methodological groundwork to rigorously test hypotheses regarding reactive electrophilic species and to develop new covalent irreversible drugs. Bioactive electrophiles such as α,β-unsaturated carbonyls are present in the environment and diet, but they are also produced endogenously during metabolism and oxidative stress. They have been linked to the etiology of diverse pathological states such as atherosclerosis, cancer, neurodegeneration, and inflammation. Understanding the roles of reactive electrophilic species in homeostasis and disease will provide important guidance for disease prevention and the development of new therapeutics. However, a lack of research tools has impeded such endeavors and many studies in this field lack experimental rigor. The problem is that these reactive electrophilic species irreversibly modify a large number of biomolecules with very little control by the scientist. The program will close critical methodological gaps in the research on the effects of electrophiles in biology. Anticipated deliverables are methods that enable the controlled generation of such electrophiles at specific times and locations. Furthermore, methods to controllably reverse the formed covalent adducts of such electrophiles will be developed. Additionally, screening technologies to develop molecules that target specific protein residues will be established. The methods are innovative because they allow answering important biomedical questions that are currently out of reach. The program’s preliminary studies have established a solid foundation to achieve these goals. We have developed chemical reactions that allow generating α,β-unsaturated carbonyls on-demand in living systems, and we have established DNA-encoded libraries for the discovery of bioactive species. With these unique capabilities in hand, the research program is in the position to advance the understanding of reactive electrophilic species in biology and their impact on disease. To demonstrate their utility, we will apply them to study the involvement of 4-hydroxynonenal in atherosclerosis and study the dynamics of adaptive states associated with resistance to KRASG12C inhibitors of pancreatic cancer. We will take steps to ensure that the developed methods will be available to a broad range of scientists to maximize the impact of the program.