Chemo- and Regio- Selective Lysine Modification on the Surface of Native Proteins: Synthetic Methods for the Improvement of Cancer Therapeutics

天然蛋白质表面的化学和区域选择性赖氨酸修饰：改进癌症治疗的合成方法

• 批准号: 10054101
• 负责人: Heemal Dhanjee
• 金额: $ 6.64万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10054101
• 关键词: Alkynes; Antibodies; Azides; Biological; Biological Testing; Biology; Chemicals; Chemistry; Communities; Complement; Coupled; Coupling; Cysteine; Development; Drug Delivery Systems; Drug Targeting; Environment; Esters; Fluorides; Generations; Goals; Imaging Techniques; Industry; Interdisciplinary Study; Ligation; Link; Lysine; Malignant Neoplasms; Manuscripts; Mediating; Mentors; Metals; Methods; Modification; Names; Occupations; Oral; Outcome; Oximes; Palladium; Peptides; Pharmacologic Substance; Physiological; Post-Translational Protein Processing; Preparation; Proteins; Protocols documentation; Pseudomonas aeruginosa toxA protein; Reagent; Reporting; Research; Scaffolding Protein; Sulfhydryl Compounds; Surface; Technology; Testing; Therapeutic; Training; Transition Elements; Trastuzumab; Universities; Variant; Work; Writing; bioimaging; experience; functional group; graduate student; hands on research; imaging biomarker; lectures; mindfulness; mutant; nanomolar; novel; post-doctoral training; scaffold; skills; small molecule; symposium; technology development; tool; trafficking; undergraduate student

PROJECT SUMMARY/ABSTRACT Protein modification has a variety of use cases, from cellular trafficking, targeted drug delivery, to imaging biomarkers. For a medicinal chemist, a single small molecule scaffold can set the stage for a nearly limitless number of controlled modifications to obtain an equally boundless set of well-defined derivatives for relevance in the context of pharmaceuticals and related industries. However, while the chemical biologist can imagine an infinite set of derivatives from a single protein scaffold, the tools available by which they may construct these bioconjugates is severely bounded therefore limiting their ability to test biological hypotheses. Moreover, the technology to enable large biomolecule coupling with stable linkages in stoichiometric quantities remains a formidable challenge and any such technology is projected to have a significant impact in the field. The preliminary results I have obtained not only enables robust, large protein-protein couplings at nanomolar concentrations, but more generally serves as a platform for rapid biomolecule diversification under biologically relevant conditions in the presence of diverse, unprotected functional groups. By using chemoselective palladium-thiol chemistry for chemoselective modification of low abundant cysteine residues, this proposal aims to expand the synthetic toolbox and chemical space available to chemical biologists. This is accomplished by the development of organometallic palladium reagents for the preparation stable, isolable, electrophilic organometallic palladium proteins for subsequent use in thiol couplings (Aims 1 and 2). To further demonstrate the utility of these organometallic palladium proteins, Aim 3 of this proposal will take advantage of this technology for the development of novel antibody-protein conjugates, an underexplored avenue of research due to the lack of technology to prepare these constructs. Thus, this proposal aims to expand the synthetic toolbox available for the chemical biologist to create a diversity of well-defined conjugates with the developed palladium reagents. Mindful of their practical use, reagents will be readily prepared, robust, and bench stable with protocols that can be easily implemented. With the goal of obtaining an academic job at a major university conducting interdisciplinary research, my training here at MIT will allow me to gain needed experience in chemical biology to complement my skills as a synthetic organic chemist. This will continue to entail hands-on research experience. In addition, I will have ample experience in writing, both in the context of proposal writing and manuscript preparation, mentoring, where I will continue to work with graduate and undergraduate students to hone this skill, and oral presentation skills by attending various conferences and guest lecturing courses. At MIT, I can continue to draw from the wealth of expertise in organometallic transformations from my colleagues in the Buchwald lab as well as that of chemical biology in the Pentelute lab making this the ideal environment for my post-doctoral training.

项目总结/摘要 蛋白质修饰有各种各样的用例，从细胞运输，靶向药物递送到成像 生物标志物。对于一个药物化学家来说，一个小分子支架可以为几乎无限的 控制修改的数量，以获得同样无限的定义明确的相关衍生物集 在制药和相关行业的背景下。然而，虽然化学生物学家可以想象一个 从一个单一的蛋白质支架衍生物的无限集，他们可以构建这些可用的工具， 生物缀合物受到严格限制，因此限制了它们测试生物学假设的能力。而且 能够使大生物分子与化学计量量的稳定键联偶联的技术仍然是一种 这是一项艰巨的挑战，预计任何此类技术都将在实地产生重大影响。的 我获得的初步结果不仅能够在纳摩尔浓度下实现强大的，大的蛋白质-蛋白质偶联， 浓度，但更一般地用作在生物学条件下快速生物分子多样化的平台。 在不同的，未保护的官能团的存在下的相关条件。 通过使用化学选择性钯-硫醇化学用于低丰度的化学选择性修饰， 半胱氨酸残基，该提案旨在扩大化学合成工具箱和化学空间， 生物学家这是通过开发用于制备的有机金属钯试剂来实现的。 稳定的、可分离的、亲电子的有机金属钯蛋白，用于随后的硫醇偶联（目的1和 2）。为了进一步证明这些有机金属钯蛋白的效用，本提案的目标3将采用 这种技术对于开发新型抗体-蛋白质缀合物的优势， 由于缺乏制备这些构建体的技术，研究途径受到限制。因此，该提案旨在扩大 化学生物学家可以使用合成工具箱来创建多种定义明确的缀合物， 开发了钯试剂。考虑到它们的实际用途，试剂将易于制备，耐用， 稳定的协议，可以很容易地实现。为了在一个专业获得一份学术工作， 作为一所进行跨学科研究的大学，我在麻省理工学院的训练将使我获得所需的经验 来补充我作为合成有机化学家的技能这将继续需要亲自动手， 研究经验。此外，我将有丰富的写作经验，无论是在提案写作方面， 和手稿准备，指导，在那里我将继续与研究生和本科生 通过参加各种会议和客座演讲课程来磨练这种技能和口头表达技能。在 麻省理工学院，我可以继续从我的同事那里汲取丰富的有机金属转化专业知识， 在Buchwald实验室以及Pentelute实验室的化学生物学实验室， 我的博士后训练