Site-Selective Modification of Peptides and Proteins through Noncovalent Interactions

项目摘要/摘要 这项拟议的研究集中在开发新的技术来进行现场选择性修饰。 基于多肽的药物。多肽已经成为有希望的替代治疗药物，但许多 缺点阻碍了它们的广泛使用。这些限制可以通过后期修改来解决 例如通过生物偶联或装订的多肽初级序列，然而选择性地 将这些化学复杂的、无保护的多肽功能化是具有挑战性的。这项提案的重点是 利用多肽的这种丰富的官能团来实现试剂和 在所需的亲核芳香取代反应(SNAR)位置附近的残留物。第一，亲电芳烃 SNAR将附加有导向基团，以与多肽上的匹配序列相互作用，指导反应性 对于邻近的半胱氨酸残基进行官能化；第二，能够进一步活化的Lewis酸性催化剂 SNAR的芳烃将被设计成同样与某些肽残基相互作用，从而实现 催化剂和配体对修饰部位的控制。这两个战略都将允许配对 具有与目标亲核性相邻的特定残基序列的特定导向基团或催化剂 残留物，如半胱氨酸。用工具箱实现多活性残基多肽的正交衍生化 可以想象，每种试剂都是针对不同环境中的亲核残基而设计的。这 建议将促进各种相关分子的快速生物偶联到多肽疗法，以 增强它们的效力，如小分子受体或药物、荧光团、其他多肽，甚至 蛋白质，如抗体。多个功能化，例如通过装订，也可以用于 提高多肽的结构稳定性和体内抗蛋白水解性。由于很少有方法 对于存在位置选择性修改的情况，这项研究将有助于研究如何对不同位置进行更改 多肽药物改变了它们的结构和功能，允许更广泛地获得更多不同的药物类似物。 用于位点选择性多肽修饰的定向反应性范例不仅对SNAR有用 反应，但代表了一种通用的方法，可以服从于许多其他标记试剂或金属- 调解的过程。这些发现将有助于实现该项目的总体目标，即 进一步发展以多肽为基础的药物成为一种强大的、备受欢迎的解决方案 当今人类健康面临的诸多挑战。麻省理工学院的潘特鲁特小组将是进行这项工作的理想地点 研究，鉴于他们在多肽合成和修饰以及多肽开发方面的专业知识 治疗学。他们高度协作和以团队为基础的实验室环境将促进思想交流， 在解决现代治疗学这一极其重要的话题方面的创造力和问题解决。

PROJECT SUMMARY/ABSTRACT This proposed research is centered on the development of new techniques for the site-selective modification of peptide-based drugs. Peptides have emerged as promising, alternative therapeutic agents, yet a number of drawbacks preclude their widespread use. These limitations can be resolved through the late-stage modification of peptide primary sequences, such as through bioconjugation or stapling, yet methods for selectively functionalizing these chemically complex, unprotected peptides is challenging. This proposal focuses on harnessing this functional group richness of peptides for noncovalent interactions between reagents and residues near sites of desired nucleophilic aromatic substitution (SNAr) reactions. First, electrophilic arenes for SNAr will be appended with directing groups to interact with matched sequences on peptides, directing reactivity toward adjacent Cys residues for functionalization Second, Lewis acidic catalysts that are able to further activate arenes for SNAr will be designed to likewise interact with certain peptide residues, enabling the achievement of catalyst- and ligand-control over sites of modification. Both of these strategies will allow for the pairing of particular directing groups or catalysts with specific sequences of residues adjacent to targeted nucleophilic residues, such as Cys. Orthogonal derivatization of peptides containing many reactive residues with a toolbox of reagents, each designed to target a nucleophilic residue in a different environment, could be imagined. This proposal will facilitate the rapid bioconjugation of a variety of relevant molecules to peptide therapeutics to enhance their efficacies, such as small molecule receptors or drugs, fluorophores, other peptides, or even proteins such as antibodies. Multiple functionalizations, such as through stapling, could also be utilized to enhance the peptides’ structural stabilities and their resistance to proteolytic cleavage in vivo. Since few methods for site-selective modification exist, this research would facilitate studies of how alterations to different locations on peptide medicines alter their structure and function, allowing for wider access to more diverse drug analogs. The paradigm of directed reactivity for site-selective peptide modifications would not only be useful for SNAr reactions, but represents a general approach amenable to a multitude of other tagging reagents or metal- mediated processes. These discoveries would serve to accomplish the overarching goal of this project, which is the further development of peptide-based medicines into a powerful, and highly sought-out approach to solve the many challenges to human health today. The Pentelute group at MIT will be an ideal location to conduct this research, given their expertise in peptide synthesis and modification and in the development of peptide therapeutics. Their highly collaborative and team-based laboratory environment will promote exchange of ideas, creativity, and problem solving in tackling the incredibly important topic of modern therapeutics.