Novel Chemical Approaches to Protein Targeting and Detection

我们的研究计划专注于使用有机/生物无机化学来开发创新的分子解决方案，从根本上解决与疾病相关的难题。这项研究可以分为两类：A)药物化学，我们寻求开发识别和抑制生物相关分子的新的化学结构；B)分子识别，我们寻求为疾病/感染的分子生物标记物开发新的诊断工具。至于建议的探知金，我们会集中研究以下课题： 1)用新型共价药物靶向电泳性反应性蛋白质组。 我们和其他人利用反应性电泳剂五氟苯磺酰胺(PFBS)来修饰目标蛋白上的亲核残基。例如，巴布林是一种含有PFBS的分子，它可以修饰B-微管蛋白秋水仙碱结合部位的半胱氨酸残基，以防止微管形成。然而，巴布林与含半胱氨酸的谷胱甘肽(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的效用并限制其对抗GSH的活性。 -一种新的亲电弹头平台：反应性光谱。我们工作的一个主要目标是创建一个弹头工具包，为每种类型的亲核分子提供选择性的反应性曲线。为此，我们在实验室开发的新化学的基础上，建议开发新的四氟苯磺酰胺类化合物，在6-位上用给电子和引出基团的光谱取代，以改变反应活性。我们的目标是探索这种新奇的化学反应，并建立一座新型弹头的武器库。 2)新型准分子启动型荧光传感器的设计与应用。 血液和脑脊液(CSF)感染的快速诊断仍然是一个高度未得到满足的临床需求。目前，由于需要培养细菌样本以进行检测，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(&lt；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，它使用传统的荧光计选择性地检测到少至10个细菌细胞。我们将对传感器进行化学优化，开发最适合信号检测的方法。

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.