Novel Chemical Approaches to Protein Targeting and Detection

我们的研究项目侧重于利用有机/生物无机化学来开发创新的分子解决方案，以解决与疾病相关的根本难题。研究可以分为两类：A)药物化学，我们寻求开发新的化学结构来识别和抑制生物学相关分子；B)分子识别，我们寻求开发新的疾病/感染分子生物标志物诊断工具。对于拟议的发现资助，我们将专注于以下研究主题：1)利用新型共价疗法靶向亲电反应性蛋白质组。我们和其他人利用活性亲电试剂五氟苯磺酰胺（PFBS）修饰靶蛋白上的亲核残基。例如，含有pfbs的Batabulin修饰b -微管蛋白秋水仙碱结合位点的Cys残基，以防止微管的形成。然而，巴蟾灵与含cys的谷胱甘肽（GSH）反应迅速，失去疗效。在这里，我们提出了一种不同的策略来挽救PFBS的效用并限制其对GSH的活性。-一种新型亲电战斗部平台：反应谱。我们工作的一个主要目标是为每种类型的亲核试剂创建一个具有选择性反应性特征的弹头工具包。为此，在本实验室发展的新化学基础上，我们建议开发新的四氟苯磺酰胺，在6位上取代具有不同反应活性的供电子和吸电子基团谱。我们的目标是探索这种新型化学物质，建立新型弹头的军械库。2)无菌液体细菌检测新型准分子荧光传感器的设计与应用。快速诊断血液和脑脊液（CSF）感染仍然是一个高度未满足的临床需求。目前，由于需要培养细菌样本才能进行检测，诊断只能在至少24小时内完成。在这里，我们的目标是开发一种新的诊断工具，用于快速（< 20分钟）检测血液和脑脊液感染。我们技术的核心是一种新型的高灵敏度荧光化学传感器，它可以使用传统的荧光计选择性地检测到10个细菌细胞。我们将对传感器进行化学优化，并开发最适合的信号检测方法。

Our research program focuses on using organic/bioinorganic chemistry to develop innovative molecular solutions to fundamentally difficult disease-related problems. The research can be placed in two categories: A) Medicinal chemistry, where we seek to develop novel chemical architectures for the recognition and inhibition of biologically relevant molecules; B) Molecular recognition, where we seek to develop novel diagnostic tools for molecular biomarkers of disease/infection. For the proposed discovery grant, we will focus on the following research topics: 1) Targeting the electrophile-reactive proteome with novel covalent therapeutics. We and others have utilized the reactive electrophile, pentafluorobenzenesulfonamide (PFBS), to modify nucleophilic residues on target proteins. For example, Batabulin, a PFBS-containing molecule modifies a Cys residue in the colchicine binding site of B-tubulin to prevent microtubule formation. However, Batabulin reacts rapidly with the Cys-containing glutathione (GSH) and loses efficacy. Here, we propose a different strategy for rescuing the utility of PFBS and limiting its activity against GSH. - A novel electrophilic warhead platform: spectrum of reactivity. A major goal of our work is to create a toolkit of warheads with selective reactivity profiles for every type of nucleophile. To this end, building on new chemistry developed in our lab, we propose to develop novel tetra-fluorobenzene sulfonamides, substituted in the 6-position with a spectrum of electron donating and withdrawing groups to vary reactivity. Our goal is to explore this novel chemistry and establish an armory of novel warheads. 2) Design and applications of novel excimer-based turn-on fluorescent sensors for bacterial detection in sterile fluids. Rapid diagnosis of infections of blood and cerebrospinal fluid (CSF) remains a highly unmet clinical need. Currently, diagnosis can only be achieved in a minimum of 24 hours, owing to the need of culturing bacteria samples to enable their detection. Here, we aim to develop a novel diagnostic tool for rapid (< 20 min) detection of blood and CSF infections. At the core of our technology is a novel class of highly sensitive fluorescent chemical sensors, which selectively detects as few as 10 bacterial cells using a conventional fluorometer. We will chemically optimize the sensors and develop the most suitable methods for signal detection.