High-Purity Peptide Libraries without Chromatographic Separation

无需色谱分离的高纯度肽文库

• 批准号: 8715569
• 负责人: KRISHNA KUMAR
• 金额: $ 22.5万
• 依托单位: VERAQUEL TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-05 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8715569
• 关键词: Acquired Immunodeficiency Syndrome; Address; Amino Acid Motifs; Amino Acids; Area; Biological; Centrifugation; Chemicals; Chromatography; Coupling; Diabetes Mellitus; Disease; Elements; Equipment; Filtration; Fluorocarbons; Fuzeon; G-Protein-Coupled Receptors; Generations; Goals; Growth; HIV; High Pressure Liquid Chromatography; Hydrophobicity; Individual; Inflammation; Isoelectric Point; Laboratories; Length; Libraries; Life; Ligand Binding; Ligands; Malignant Neoplasms; Marketing; Methods; Metric; Modification; Non-Insulin-Dependent Diabetes Mellitus; Organic solvent product; Patients; Peptide Library; Peptide Synthesis; Peptides; Pharmaceutical Preparations; Phase; Physical condensation; Process; Proteins; Reagent; Research; Sales; Scientist; Side; Site; Solid; Solubility; Solutions; T-20; Technology; Testing; Therapeutic; Training; Variant; Water; Work; analog; biophysical properties; cost; design; empowered; exenatide; experience; glucagon-like peptide 1; innovation; large scale production; manufacturing process; member; milligram; novel; prevent; programs; public health relevance; success

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.

描述（由申请人提供）：肽和蛋白质产品的市场每年达 300 亿美元，并且仍在持续增长。自动化固相肽合成 (SPPS) 是现代医学研究的支柱，已应用于微摩尔至公吨的工业规模。如果没有 SPPS，诸如艾塞那肽（GLP-1 类似物，许多 2 型糖尿病患者的救星）和治疗 HIV-AIDS 的 Fuzeon 等肽类药物就不会存在。不完整的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。我们的技术通过在后台（标准缩合步骤之间）进行化学校对，为该问题提供了一种新颖的解决方案，无需对现有方法进行修改。几分钟之内，任何未反应的位点都会被惰性碳氟化合物残留物覆盖。因此，错误序列被减少为不溶性构建体，最终可以轻松地去除。我们的初步研究始终提供分离的全长肽，特别是在之前经过重复 HPLC 纯化的情况下。我们的目标是将这项技术扩展到肽库的无差错合成。达到高纯度将建立 肽研究的新授权标准。我们塔夫茨大学的研究项目在 G 蛋白偶联受体 (GPCR) 的研究方面拥有丰富的经验，并在新型肽配体的设计和合成方面产生了显着的创新。因此，我们建议合成 GLP-1 类似物文库，体现新颖且非天然的氨基酸基序。总而言之，这些研究将实现纯度超过 70% 的肽库的高效合成，无需通过色谱法纯化，并可能在项目期结束时产生治疗糖尿病的相关化合物。