Cysteine Arylation

描述（由申请人提供）：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，鉴于现有方法的固有局限性，需要新的化学工具来进行生物分子的常规修饰和工程。具体来说，在肽水平上，最近的进展包括使用大环化化学通过碳氢化合物连接物、芳香夹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，引入的约束导致多肽能够穿透细胞，从而使这些变体可以靶向大范围的细胞内蛋白质和相互作用。最近化学应用于蛋白质疗法的例子包括使用总蛋白合成来制备靶向VEGF的镜像蛋白，将非天然氨基酸结合用于peg样聚合物与蛋白质药物的位点特异性附着，以及小分子抗体治疗偶联物。目前，人们正在努力设计肽和蛋白质作为医学相关蛋白质相互作用的抑制剂；这些分子正被广泛开发，因为它们具有很大的表面积，可以结合和识别具有高亲和力和特异性的目标。无保护肽和蛋白质的化学修饰被证明是具有挑战性的，因为一个位点经常需要在许多其他官能团的存在下进行标记或修饰。因此，所采用的反应需要具有化学选择性，区域选择性，并在温和的生物分子“友好”条件下操作。在这里，我们建议发展半胱氨酸芳基化作为生物偶联的广泛工具，并破译这些全氟芳基如何影响一类生物活性肽和蛋白质的结构和功能。我们的目标是用各种全氟芳基试剂研究半胱氨酸芳基化，开发酶催化的化学反应，并对含有这些全氟芳基片段的生物活性蛋白和肽进行结构-功能研究。我们认为，这里提出的研究将通过引入一种新的方法来修饰肽和蛋白质，为化学生物学领域做出基础和实际的贡献，并将影响下一代生物治疗学领域。

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.