Novel Chemical Approaches to Protein Targeting and Detection

我们的研究计划侧重于使用有机/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。该研究可以分为两类：A）药物化学，我们寻求开发新的化学结构，用于识别和抑制生物相关分子; B）分子识别，我们寻求开发新的诊断工具，用于疾病/感染的分子生物标志物。对于拟议的发现资助，我们将专注于以下研究课题：1）以新型共价疗法靶向亲电反应蛋白质组。 我们和其他人已经利用反应性亲电试剂五氟苯磺酰胺（PFBS）来修饰靶蛋白上的亲核残基。例如，Batabulin，一种含有PFBS的分子，修饰B-微管蛋白的秋水仙素结合位点中的Cys残基以防止微管形成。然而，Batabulin与含Cys的谷胱甘肽（GSH）迅速反应并失去功效。在这里，我们提出了一种不同的策略，拯救PFBS的效用和限制其对GSH的活性。- 一种新型亲电子弹头平台：反应谱。我们工作的一个主要目标是创建一个工具包的弹头具有选择性的反应性档案的每一种类型的亲核试剂。为此，在我们实验室开发的新化学的基础上，我们建议开发新的四氟苯磺酰胺，在6-位上取代，具有供电子和吸电子基团的光谱以改变反应性。我们的目标是探索这种新型化学物质并建立新型弹头军械库。2)新型准分子开启荧光传感器在无菌液体细菌检测中的设计与应用。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，因此最少只能在24小时内进行诊断。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CSF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.