Cysteine Arylation

描述(申请人提供)：应用于多肽和蛋白质的精细化学工具导致了无与伦比的结构-功能研究和量身定制的生物疗法。然而，考虑到现有方法的固有局限性，需要新的化学工具来进行常规的生物分子修饰和工程。具体地说，在多肽水平上，最近取得进展的领域包括使用大环化化学通过碳氢连接物、芳香族夹子或内酰胺桥来限制这些物种。这些修饰在多肽治疗领域引起了兴奋，因为在某些情况下，引入的限制导致了能够穿透细胞的多肽，从而使这些变体可以针对广泛的细胞内蛋白质和相互作用。最近应用于创建蛋白质疗法的化学例子包括：利用总蛋白质合成来制备靶向血管内皮生长因子的镜像蛋白质，将非天然氨基酸用于聚乙二醇类聚合物与蛋白质药物的定点结合，以及小分子抗体治疗性结合物。目前，设计多肽和蛋白质作为医学上相关的蛋白质相互作用的抑制剂的工作正在进行中；这些分子正在被广泛开发，因为它们显示出较大的表面积，可以结合和识别具有高亲和力和特异性的靶标。事实证明，对未受保护的多肽和蛋白质进行化学修饰具有挑战性，因为一个位点经常需要在许多其他官能团存在的情况下进行标记或修饰。因此，所采用的反应必须是化学选择性的、区域选择性的，并且在温和的生物分子“友好”条件下可操作。在这里，我们建议开发半胱氨酸芳基化作为一种广泛的生物偶联工具，并破译这些全氟芳基如何影响一类生物活性多肽和蛋白质的结构和功能。我们的目标是研究半胱氨酸与各种全氟芳香族试剂的芳基化反应，开发酶催化版本的化学，并对含有这些全氟芳基的生物活性蛋白质和多肽进行结构和功能研究。我们认为，本文提出的研究将通过引入一种新的多肽和蛋白质修饰方法，为化学生物学领域做出基础性和实用性的贡献，并将对下一代生物治疗领域产生影响。

DESCRIPTION (provided by applicant): Elaborate chemistry tools applied to peptides and proteins have led to unparalleled structure-function studies and tailor-made bio-therapeutics. Yet, given inherent limitations of existing methods new chemical tools are needed for routine modification and engineering of biomolecules. Specifically, at the peptide level, areas of recent advancement include the use of macrocyclization chemistry to constrain these species via hydrocarbon linkers, aromatic clips, or lactam bridges. These modifications have generated excitement in the field of peptide therapeutics because in some cases the introduced constraint has led to peptides capable of penetrating cells thereby making a wide range of intracellular proteins and interactions targetable by these variants. Recent examples for chemistry applied to the creation of protein therapeutics involve the use of total protein synthesis to prepare mirror image proteins that target VEGF, the incorporation of non-natural amino acids for site-specific attachment of PEG-like polymers to protein drugs, and the small molecule antibody therapeutic conjugates. Currently, significant efforts are underway to design peptides and proteins as inhibitors of medically relevant protein-protein interactions; these molecules are being widely developed because they display a large surface area that can bind and recognize a target with high affinity and specificity. The chemical modification of unprotected peptides and proteins has proved challenging because one-site often needs to be labeled or modified in the presence of many other functional groups. Consequently, the reactions employed need to be chemoselective, regioselective, and operational under mild, biomolecule "friendly" conditions. Here we propose to develop cysteine arylation as an extensive tool for bioconjugation and to decipher how these perfluoroaryl groups affect the structure and function of a select class of bioactive peptides and proteins. We aim to investigate cysteine arylation with a variety of perfluoroaromatic reagents, to develop enzyme-catalyzed version of the chemistry, and to carry out structure-function studies on bioactive proteins and peptides containing these perfluoroaryl moieties. In our opinion, the research proposed here will make fundamental and practical contributions to the field of chemical biology by introducing a new method to modify peptides and proteins, and will impact the area of next-generation bio-therapeutics.