High-Purity Peptide Libraries without Chromatographic Separation

描述（由申请人提供）：肽和蛋白质产品的市场每年为300亿美元，并继续增长。自动化固相肽合成（SPPS）是现代药物研究的支柱，并已用于微摩尔到公吨的工业规模。肽类药物，如Exengland，GLP-1类似物和许多2型糖尿病患者的救命药，以及用于HIV-AIDS的Fuzeon，如果没有SPPS，就不会存在。由不完全合成步骤导致的错误序列限制了SPPS的效率和通量，通常需要费力且昂贵的分离。我们的技术通过在背景中（标准缩合步骤之间）进行化学校对，为该问题提供了一种新的解决方案，无需对现有方法进行修改。在几分钟内，任何未反应的位点都被惰性碳氟化合物残基覆盖。因此，错误序列被还原成不溶性的构建体，其可以在最后以容易的方式去除。我们的初步研究一致地提供了分离的全长肽，特别是在先前进行重复HPLC纯化的情况下。我们的目标是将这一技术扩展到合成无错合成肽库。达到高纯度将建立 肽研究的新授权标准。我们在塔夫茨大学的研究项目在G蛋白偶联受体（GPCR）的研究方面有经验，并在新型肽配体的设计和合成方面取得了显着的创新。因此，我们提出GLP-1类似物的库的合成，体现新的和非天然的氨基酸基序。总之，这些研究将导致纯度超过70%的肽的有效文库合成，而不需要通过色谱法纯化，并且可能在项目期结束时产生治疗糖尿病的相关化合物。

DESCRIPTION (provided by applicant): The market for peptide and protein products is $30 billion annually and continues to grow. Automated solid phase peptide synthesis (SPPS) is a mainstay of modern medicinal research, and has been used on micromole up to metric ton industrial scale. Peptide drugs such as Exenatide, a GLP-1 analogue and a life-saver to many type-2 diabetes patients, and Fuzeon for HIV-AIDS would not exist but for SPPS. Error sequences resulting from incomplete synthetic steps limit the efficiency and throughput of SPPS, often requiring laborious and costly separations. Our technology offers a novel solution to the problem via a chemical proofreading that takes place in the background (between standard condensation steps) with no modification to established methods. Within minutes, any unreacted sites are capped with inert fluorocarbon residues. The error sequences are thus reduced into insoluble constructs that can be removed in a facile manner at the end. Our preliminary studies have consistently afforded isolated full- length peptides, and especially in cases previously subject to repeated HPLC purification. We aim to expand this technology to the synthesis of error-free synthesis of peptide libraries. Achieving high purities would establish a new empowering standard in peptide research. Our research program at Tufts has experience in the study of G- Protein coupled receptors (GPCRs) and has produced notable innovations in the design and synthesis of novel peptide ligands. We therefore propose the synthesis of libraries of GLP-1 analogs, embodying novel and unnatural amino-acid motifs. Taken together, these studies will result in efficient library synthesis of peptides with purities exceeding 70% without the need for purification by chromatography and may result in a therapeutically relevant compound for diabetes at the end of the project period.