Cysteine Arylation

描述（由申请人提供）：应用于肽和蛋白质的精细化学工具已经导致了无与伦比的结构-功能研究和定制的生物疗法。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化。具体而言，在肽水平上，最近的进展领域包括使用大环化化学通过烃接头、芳族夹或内酰胺桥来限制这些物质。这些修饰在肽治疗领域中产生了兴奋，因为在某些情况下，引入的限制导致肽能够穿透细胞，从而使广泛的细胞内蛋白质和相互作用可被这些变体靶向。应用于产生蛋白质治疗剂的化学的最新实例涉及使用总蛋白质合成来制备靶向VEGF的镜像蛋白质，掺入非天然氨基酸用于PEG样聚合物与蛋白质药物的位点特异性连接，以及小分子抗体治疗性缀合物。目前，正在进行大量的努力来设计肽和蛋白质作为医学相关蛋白质-蛋白质相互作用的抑制剂;这些分子正在被广泛开发，因为它们显示出可以以高亲和力和特异性结合和识别靶标的大表面积。未经保护的肽和蛋白质的化学修饰已被证明是具有挑战性的，因为一个位点通常需要在许多其他官能团的存在下被标记或修饰。因此，所采用的反应需要是化学选择性的，区域选择性的，并且在温和的生物分子“友好”条件下操作。在这里，我们建议开发半胱氨酸芳基化作为一个广泛的工具，生物共轭和破译这些全氟芳基基团如何影响一类选择的生物活性肽和蛋白质的结构和功能。我们的目标是调查半胱氨酸芳基化与各种全氟芳香族试剂，开发酶催化的化学版本，并进行生物活性的蛋白质和肽的结构-功能研究，含有这些全氟芳基部分。我们认为，本文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性和实用性的贡献，并将影响下一代生物治疗领域。

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.