Fmoc-Based Synthesis of Disulfide-Rich Cyclic Peptides
Fmoc-Based Synthesis of Disulfide-Rich Cyclic Peptides
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DOI：
10.1021/jo500699m
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发表时间：
2014-06-20
影响因子：
3.6
通讯作者：
Craik, David J.
中科院分区：
文献类型：
--
作者：
Cheneval, Olivier;Schroeder, Christina I.;Craik, David J.
富含二硫键的环肽作为药物发现的先导或框架具有令人兴奋的潜力;然而，它们的使用面临着一些合成挑战，主要与环状骨架的构建和正确二硫键的形成有关。在这里，我们描述了一个简单而有效的Fmoc固相肽合成（SPPS）为基础的方法合成富含二硫键的环肽。该方法包括在2-氯三苯甲基树脂上的SPPS，在溶液中部分保护的肽的环化，侧链保护基团的裂解，以及半胱氨酸的氧化以产生所需产物。我们用三种不同类型的含环胱氨酸结基序的环肽合成肽来说明这种方法：Mobius（M），胰蛋白酶抑制剂（T）和手镯（B）。我们表明，该方法是广泛适用于肽工程，说明了两个突变体和三个接枝类似物的kalata B1的合成。该方法减少了高腐蚀性和毒性试剂的使用，并且比先前报道的用于生产富含二硫化物的环肽的方法更适合于高通量合成，从而提供了促进这些支架的药物优化的巨大潜力。
Disulfide-rich cyclic peptides have exciting potential as leads or frameworks in drug discovery; however, their use is faced with some synthetic challenges, mainly associated with construction of the circular backbone and formation of the correct disulfides. Here we describe a simple and efficient Fmoc solid-phase peptide synthesis (SPPS)-based method for synthesizing disulfide-rich cyclic peptides. This approach involves SPPS on 2-chlorotrityl resin, cyclization of the partially protected peptide in solution, cleavage of the side-chain protecting groups, and oxidization of cysteines to yield the desired product. We illustrate this method with the synthesis of peptides from three different classes of cyclic cystine knot motif-containing cyclotides: Mobius (M), trypsin inhibitor (T), and bracelet (B). We show that the method is broadly applicable to peptide engineering, illustrated by the synthesis of two mutants and three grafted analogues of kalata B1. The method reduces the use of highly caustic and toxic reagents and is better suited for high-throughput synthesis than previously reported methods for producing disulfide-rich cyclic peptides, thus offering great potential to facilitate pharmaceutical optimization of these scaffolds.