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。对于拟议的发现基金，我们将专注于以下研究主题：*1)以新型共价疗法为靶点的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的化学物质，并建立一个新的弹头武器库。*2)基于准分子的新型开启荧光传感器的设计和应用，用于无菌液体中的细菌检测。*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.