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(申请人提供)：多肽和蛋白质产品的市场规模为每年300亿美元，而且还在持续增长。固相肽的自动化合成是现代医学研究的支柱，已在微摩尔到吨级的工业规模上得到了应用。多肽药物，如埃克塞那肽，一种GLP-1类似物，是许多2型糖尿病患者的救命稻草，以及治疗艾滋病毒-艾滋病的Fuzeon，如果没有SPPS，就不会存在。合成步骤不完整导致的错误序列限制了SPP的效率和吞吐量，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)进行的化学校对为该问题提供了一种新的解决方案，而不需要对现有方法进行任何修改。在几分钟内，任何未反应的部位都会被惰性的氟碳残留物覆盖。因此，错误序列被简化为不能溶解的结构，可以在结束时以容易的方式移除。我们的初步研究一致地提供了分离的全长多肽，特别是在以前需要重复进行高效液相纯化的病例中。我们的目标是将这一技术扩展到无误合成多肽库的合成中。达到高纯度将建立 多肽研究中的一个新赋权标准。我们在塔夫茨的研究项目在G蛋白偶联受体(GPCRs)的研究方面拥有经验，并在设计和合成新型多肽配体方面取得了显着的创新。因此，我们建议合成GLP-1类似物的文库，包含新的和非自然的氨基酸基序。综上所述，这些研究将导致有效地合成纯度超过70%的多肽，而不需要通过层析进行纯化，并可能在项目期结束时产生对糖尿病具有治疗意义的化合物。