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

未经修改，来自原始应用程序。根据授予1F32GM131592-01A1 项目摘要/摘要 蛋白质修饰有多种用途，从细胞运输、靶向药物输送到成像 生物标志物。对于药物化学家来说，一个单一的小分子支架就可以为几乎无限的 受控修改的数量，以获得同样无穷无尽的明确定义的相关性衍生工具集 在制药和相关行业的背景下。然而，当化学生物学家可以想象一个 来自单个蛋白质支架的无限组衍生品，他们可以用来构建这些衍生品的工具 生物偶联物受到严格的限制，因此限制了它们检验生物假说的能力。此外， 能够以化学计量比实现具有稳定连接的大生物分子偶联的技术仍然是 这是一项艰巨的挑战，任何此类技术都预计将在该领域产生重大影响。这个 我所获得的初步结果不仅实现了纳摩尔的强健、大蛋白质-蛋白质偶联 浓缩，但更普遍地作为一个平台，快速生物分子多样化在生物学上 在存在不同的、不受保护的官能团的情况下，可能会出现相关的情况。 用化学选择性钯-硫醇化学对低丰度进行化学选择性改性 半胱氨酸残基，这项建议旨在扩大合成工具箱和化学可用化学空间 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、亲电的有机金属钯蛋白，用于随后的硫醇偶联(目标1)。至 进一步证明这些有机金属钯蛋白的效用，本提案的目标2将采取 这项技术在开发新型抗体-蛋白质结合物方面的优势尚未得到探索 由于缺乏准备这些结构的技术，研究的道路。因此，这项提案旨在扩大 为化学生物学家提供的合成工具箱，可用于创建各种定义明确的与 开发了钯试剂。考虑到它们的实际用途，试剂将随时准备好，坚固耐用 协议稳定，易于实施。目标是在一个专业获得一份学术工作 大学进行跨学科研究，我在麻省理工学院的培训将使我获得所需的经验 化学生物学来补充我作为合成有机化学家的技能。这将继续需要亲身实践 研究经验。此外，我将在写作方面有丰富的经验，无论是在提案撰写的背景下 以及手稿准备和指导，在那里我将继续与研究生和本科生合作 通过参加各种会议和客座演讲课程，磨练这项技能和口头表达能力。在… 麻省理工学院，我可以继续借鉴我的同事们在有机金属转化方面的丰富专业知识 布赫瓦尔德实验室和潘特鲁特实验室的化学生物学使这里成为 我的博士后培训。

Unmodified, from original application. Awarded under Grant 1F32GM131592-01A1 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 (Aim 1). To further demonstrate the utility of these organometallic palladium proteins, Aim 2 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.