IMPROVED HANDLES FOR SOLID-PHASE PEPTIDE SYNTHESIS

固相肽合成的改进方法

• 批准号: 3301546
• 负责人: George Barany
• 金额: $ 14.83万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3301546
• 关键词: amides; asparagine; branched chain aminoacid; carboxyl group; chemical cleavage; chemical reaction; chemical stability; crosslink; glutamine; peptide chemical synthesis; peptides; polymers; solid state

The solid-phase method introduced by Merrifield is now firmly established as a powerful technique for the study of biologically important peptides. Synthesis normally begins by covalently liking the C-terminal amino acid residue of the desired peptide to an insoluble polymeric support. This anchoring step is an integral part of the overall synthetic plan, and the experimental details can impact significantly on the overall purity and yield of the final product. As part of our general interest in achieving milder chemical methods for peptide synthesis, we have had several occasions to pursue the advantages of "handle" approaches to anchoring. Handles are defined as bifunctional spacers which serve to attach the initial residue to the polymeric support in two discrete steps. They allow precise control over the stability and ultimate cleavage of the anchoring linkage, and facilitate quantitative attachments which circumvent problems associated with extraneous polymer-bound functional groups. Handles developed in our laboratory are compatible with readily removable N alpha-amino protecting groups such as the highly acid-labile N alpha-biphenylyl- isopropyloxycarbonyl (Bpoc), the base-labile N alpha-9-fluorenylmethyloxycarbonyl (Fmoc), and the thiolysable N alpha-dithiasuccinoyl (Dts) or N alpha-3-nitro-2-pyridine-sulfenyl (Npys) functions. Cleavages of the anchoring linkages occur under relatively mild conditions with acid, light, or fluoride ion; the products may be peptide acids or amides. Application of these handles in orthogonal protection schemes offers the further possibility of preparing partially protected segment suitable for purification followed by restitching to make longer and more complicated products. The present proposal aims to take several handles that have already been worked out in simple systems and demonstrate their scope when applied to challenging peptide targets. Simultaneously, some modified and new chemistries are suggested that may lead to new handles with even better properties and/or a wider range of applications.

Merrifield 推出的固相方法现已得到广泛认可 被确立为生物学研究的强大技术 重要的肽。 合成通常从共价喜欢开始 将所需肽的 C 端氨基酸残基转化为不溶性的 聚合物支撑。 该锚定步骤是 总体合成计划，实验细节会影响 显着影响最终产品的整体纯度和收率。 作为我们对实现更温和的化学方法的普遍兴趣的一部分 肽合成，我们曾多次追求 “手柄”锚定方法的优点。 句柄定义为 双功能间隔基，用于将初始残基连接到 聚合物支撑分两个独立的步骤进行。 它们允许精确控制 锚定连接的稳定性和最终裂解，以及 促进定量附件，从而避免相关问题 具有外来聚合物结合的官能团。 手柄开发于 我们的实验室与易于去除的 N α-氨基兼容 保护基团，例如高度酸不稳定的 N α-联苯基- 异丙氧基羰基 (Bpoc)，对碱不稳定 N α-9-芴基甲氧基羰基 (Fmoc)，以及可硫解的 N α-二硫代琥珀酰基 (Dts) 或 N α-3-硝基-2-吡啶-硫基 (Npys) 函数。 锚定连接的断裂发生在 相对温和的酸、光或氟离子条件；这 产品可以是肽酸或酰胺。 这些句柄的应用 在正交保护方案中提供了进一步的可能性 制备适合随后纯化的部分受保护的片段 通过重新缝合来制造更长、更复杂的产品。 目前的提案旨在采用已经存在的几个句柄 在简单的系统中进行计算并展示其应用范围 具有挑战性的肽靶标。 同时，一些修改和新增 建议使用化学物质，可能会产生性能更好的新手柄 属性和/或更广泛的应用。