IMPROVED HANDLES FOR SOLID-PHASE PEPTIDE SYNTHESIS

由梅里菲尔德引进的固相法现在已牢固地建立起来 作为研究生物重要肽的有力技术。 合成通常通过共价连接C-末端氨基酸开始 将所需肽的残基转移到不溶性聚合物载体上。 这 锚定步骤是整个综合计划的组成部分， 实验细节可显著影响总纯度， 最终产品的产量。 手柄被定义为双功能间隔物， 或连接体，其用于在两个连接体中实现所需的连接。 离散的化学步骤。手柄方法允许精确控制 锚定连接的稳定性和最终断裂，以及 便于定量连接， 与外来的聚合物结合的官能团。 主持下 当前授权，一些句柄（PAL，HAL，XAL，NPE，Nonb等）有 发展并应用于分步或分段缩合 挑战性肽靶标的合成。 相容性N-α-氨基 保护主要由酸不稳定的叔丁氧羰基提供 (Boc)或碱不稳定的9-芴基甲氧基羰基（Fmoc）官能团，和 最后的分裂发生在相对温和的条件下， 暴露于酸、碱、光、氟离子或钯时的反应 (O)在亲核试剂的存在下 根据实验 设计，产物可以是肽酸或酰胺，或者完全 游离或保留正交侧链保护。 一些 在这项研究计划中开发的试剂和程序已经实现了 在世界各地广泛使用。 目前的更新申请旨在扩大这一进展， 继续演示上述手柄如何帮助解决 在合成肽化学中的生物学显著挑战。 与此同时，正在研究改进的和新的化学物质， 导致具有更好性能和/或更宽范围的 用途，包括制备用于组织靶向的肽缀合物， 交付. 这些方法将适用于一系列尺度， 与药物发现计划和疾病诊断程序兼容 基于多肽合成。

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 linking 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. Handles are defined as bifunctional spacers, or linkers, which serve to achieve the required attachment in two discrete chemical steps. The handle approach allows precise control over the stability and ultimate cleavage of the anchoring linkage, and facilitates quantitative attachments which circumvent problems associated with extraneous polymer-bound functional groups. Under the aegis of the present grant, a number of handles (PAL, HAL, XAL, NPE, Nonb, etc.) have been developed and applied to the stepwise or segment condensation synthesis of challenging peptide targets. Compatible N-alpha-amino protection is provided principally by acid-labile tert-butyloxycarbonyl (Boc) or base-labile 9-fluorenylmethyloxycarbonyl (Fmoc) functions, and final cleavages occur under relatively mild conditions with minimal side reactions upon exposure to acid, base, light, fluoride ion, or palladium (O) in the presence of nucleophiles. Depending on the experimental design, the products can be peptide acids or amides, either completely free or else retaining orthogonal side-chain protection. Some of the reagents and procedures developed in this research program have achieved widespread use throughout the world. The present renewal application aims to expand on this progress and continue to demonstrate how the aforementioned handles can help address biologically significant challenges in synthetic peptide chemistry. Simultaneously, modified and new chemistries are being examined that may lead to new handles with even better properties and/or a wider range of uses, including the preparation of peptide conjugates for tissue-targeted delivery. These methods will accommodate a range of scales, and be compatible with drug discovery programs and disease diagnosis procedures based on multiple peptide synthesis.