Exploiting protein stability changes to identify next generation chemical probes and target engagement technologies

利用蛋白质稳定性变化来识别下一代化学探针和目标接合技术

• 批准号: RGPIN-2022-03107
• 负责人: Page, Brent
• 金额: $ 2.7万
• 依托单位: University of British Columbia
• 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=762008
• 关键词: Exploiting; protein; stability; changes; identify

Background  My research focusses on cutting-edge chemical biology and medicinal chemistry projects that prioritize the concept of "target engagement" - the direct physical interaction of chemical probes with their cellular target. Major aims within this program include developing innovative target engagement technologies and using them to study interactions between chemical probes and their target proteins. Additionally, this program includes an innovative new approach to produce targeted protein degraders (proteolysis targeting chimeras, PROTACs). Objectives and Approach Long-term goal: My research program aims to produce target engagement technologies and PROTACs that are used to explore complex biological phenomena. These tools will allow us to develop a better understanding of our target proteins, such as STAT1, STAT3, CNOT3 and TK1. Further, our novel target engagement technologies will have far-reaching effects within chemical probe discovery, facilitating the development of future generations of chemical probes. Short-term Objectives: 1.Optimize protocols for producing target engagement biosensors within our CeTEAM platform My research team has generated a novel target engagement technology called "CeTEAM" - Cellular Target Engagement by Accumulation of Mutant. This technique uses a biosensor approach for target engagement that is high-throughput and does not require an external stimulus, such as heat. We have demonstrated that CeTEAM biosensors can effectively indicate if an endogenous target protein is engaged by a chemical probe. Within this objective we will expand our repertoire of CeTEAM biosensors and focus on optimizing our process for CeTEAM biosensor development. 2.Develop novel target engagement assays that exploit unique aspects of E. coli biology To improve throughput and reduce barriers to implementation, we have turned to non-mammalian systems for investigating cellular target engagement. In this objective we aim to exploit unique aspects of E. coli biology to produce a novel target engagement technique called LIBR-TEA (ligand-induced Inclusion Body Rescue - Target Engagement Assay, "liberty"). 3. Exploit small biomolecules to produce targeted degraders as chemical biology tools As a highly innovative approach to PROTACs, we will employ naturally occurring biomolecules to target our PROTACs to proteins of interest. As a proof of concept study, we have developed thymidine-inspired PROTACs to target thymidine kinase 1 (TK1) and demonstrated that they can reduce TK1 protein levels in cells. This objective will focus on optimizing bio-inspired PROTACs for TK1 and other targets.  Impact Our novel target engagement techniques and new approach to PROTAC development is set to have a transformative impact on the chemical biology field. These tools will allow us to develop a better understanding of complex biological systems and will facilitate the discovery and validation of future chemical probes and targeted degraders.

我的研究重点是前沿化学生物学和药物化学项目，优先考虑“靶标参与”的概念-化学探针与其细胞靶标的直接物理相互作用。该项目的主要目标包括开发创新的靶标接合技术，并利用它们来研究化学探针与其靶蛋白之间的相互作用。此外，该计划还包括一种创新的新方法来生产靶向蛋白质降解物（靶向嵌合体蛋白水解，PROTACs）。长期目标：我的研究项目旨在生产用于探索复杂生物现象的目标接合技术和protac。这些工具将使我们能够更好地了解我们的靶蛋白，如STAT1， STAT3， CNOT3和TK1。此外，我们的新目标接合技术将对化学探针的发现产生深远的影响，促进未来几代化学探针的发展。短期目标：我的研究团队已经产生了一种新的目标交战技术，称为“CeTEAM”——通过突变体积累的细胞目标交战。该技术使用生物传感器方法进行高通量目标接触，不需要外部刺激，如热。我们已经证明CeTEAM生物传感器可以有效地指示内源性靶蛋白是否被化学探针参与。在此目标下，我们将扩大CeTEAM生物传感器的范围，并专注于优化CeTEAM生物传感器的开发过程。2.开发利用大肠杆菌生物学独特方面的新型靶标结合分析，为了提高通量并减少实施障碍，我们已经转向非哺乳动物系统来研究细胞靶标结合。在这个目标中，我们的目标是利用大肠杆菌生物学的独特方面来产生一种新的靶标结合技术，称为lib - tea（配体诱导包涵体拯救-靶标结合测定，“自由”）。3. 作为一种高度创新的PROTACs方法，我们将利用天然存在的生物分子将我们的PROTACs靶向到感兴趣的蛋白质上。作为概念验证研究，我们开发了胸腺嘧啶激发的PROTACs靶向胸腺嘧啶激酶1 (TK1)，并证明它们可以降低细胞中TK1蛋白的水平。该目标将集中于优化生物启发的PROTACs用于TK1和其他靶点。我们的新型靶标接合技术和PROTAC开发的新方法将对化学生物学领域产生变革性影响。这些工具将使我们能够更好地理解复杂的生物系统，并将促进未来化学探针和靶向降解剂的发现和验证。