Novel Chemical Approaches to Protein Targeting and Detection

蛋白质靶向和检测的新化学方法

• 批准号: RGPIN-2019-07109
• 负责人: Gunning, Patrick
• 金额: $ 4.66万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=746967
• 关键词: Novel; Chemical; Approaches; Protein; Targeting

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.

我们的研究计划着重于使用有机/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困难问题。这项研究可以分为两类：a）药物化学，我们试图开发新型的化学体系结构，以识别和抑制生物学相关的分子； b）分子识别，我们试图开发用于疾病/感染分子生物标志物的新型诊断工具。对于拟议的发现赠款，我们将重点介绍以下研究主题：1）针对新型共价疗法的电力反应蛋白质组。 我们和其他人利用了反应性电力，五氟苯甲苯甲酰胺（PFBS）来修饰靶蛋白上的亲核残基。例如，含PFBS的分子巴达布蛋白修饰了B-微管蛋白的秋水仙碱结合位点的Cys残基，以防止微管形成。然而，巴达布蛋白与含Cys的谷胱甘肽（GSH）反应迅速并失去疗效。在这里，我们提出了一种不同的策略，以挽救PFB的实用性并限制其对GSH的活动。 - 一种新型的亲电弹头平台：反应性频谱。我们工作的主要目标是为每种类型的亲核器创建具有选择性反应性概况的弹头工具包。为此，基于我们实验室开发的新化学反应的建设，我们建议开发新型的四氟苯磺酰胺，在6位中取代，并用一系列电子捐赠和撤回基团以改变反应性。我们的目标是探索这种新颖的化学反应并建立一支新颖的弹头军械库。 2）在无菌流体中，新型基于Impimer的转交荧光传感器的设计和应用。快速诊断出血液和脑脊液（CSF）的感染仍然是高度未满足的临床需求。目前，由于需要培养细菌样品才能检测到诊断，才能至少在24小时内实现诊断。在这里，我们旨在开发一种新型的诊断工具，以快速（<20分钟）对血液和CSF感染的检测。我们技术的核心是一类新型的高度敏感荧光化学传感器，使用常规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，并开发最合适的信号检测方法。