IMPROVED HANDLES FOR SOLID-PHASE PEPTIDE SYNTHESIS

梅里菲尔德提出的固相法现已确立。 作为研究生物重要多肽的一种强有力的技术。 合成通常是通过共价连接C端氨基酸开始的 将所需的多肽残基连接到不溶的聚合物载体上。这 锚定步骤是整体综合计划的组成部分，而 实验细节可以显著影响总体纯度和 最终产品的产量。把手定义为双功能间隔物， 或链接器，这些链接器用于在两个 离散的化学步骤。手柄方法允许精确控制 锚定连杆的稳定性和最终劈裂，以及 促进量化附件，从而避免相关问题 具有无关的聚合物结合官能团。在美国政府的支持下 当前授权、多个句柄(PAL、HAL、XAL、NPE、NONB等)有 已开发并应用于分段冷凝或分段冷凝 具有挑战性的多肽靶标的合成。相容的N-α-氨基 保护主要由酸不稳定的叔丁氧基提供。 (BOC)或不稳定碱的9-荧甲氧羰基(FMOC)功能，以及 最终的卵裂发生在相对温和的条件下，侧边最小。 接触酸、碱、光、氟离子或钯时的反应 (O)在亲核分子在场的情况下。取决于实验中的 设计，该产品可以是多肽酸或酰胺，或完全 自由或保留正交侧链保护。其中一些 在这个研究项目中开发的试剂和程序已经实现了 在世界各地广泛使用。 本续期申请旨在扩大这一进展，并 继续演示上述句柄如何帮助解决 合成肽化学在生物学上的重大挑战。 同时，正在检查改进的和新的化学物质，这些化学物质可能 导致新的手柄具有更好的性能和/或更广泛的 用途，包括制备组织靶向的多肽结合物 送货。这些方法将适应一系列规模，并将 与药物发现计划和疾病诊断程序兼容 基于多肽的合成。

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.