Phage-encoded combinatorial chemical libraries based on bicyclic peptides
Phage-encoded combinatorial chemical libraries based on bicyclic peptides
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DOI：
10.1038/nchembio.184
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发表时间：
2009-07-01
影响因子：
14.8
通讯作者：
Winter, Greg
中科院分区：
文献类型：
--
作者：
Heinis, Christian;Rutherford, Trevor;Winter, Greg
在这里，我们描述了一种噬菌体策略，用于选择基于共价连接到有机核心的双环或线性肽的配体。我们设计了具有三个反应性半胱氨酸残基的肽库，每个残基由几个随机氨基酸残基隔开，并且我们将这些肽库融合到噬菌体基因-3-蛋白。通过反应性半胱氨酸与三-（溴甲基）苯缀合产生具有锚定至均三甲苯核心的两个肽环的肽缀合物的库。迭代亲和选择产生了几种酶抑制剂;在进一步诱变和选择后，我们能够化学合成对人血浆激肽释放酶特异性的先导抑制剂（PK 15; K（i）= 1.5 nM），其有效地中断了离体测试的人血浆中的固有凝血途径。该方法提供了产生和选择双环大环（或如果裂解，其线性衍生物）作为在小分子药物和生物制剂的界面处稳定的配体的有力手段。
Here we describe a phage strategy for the selection of ligands based on bicyclic or linear peptides attached covalently to an organic core. We designed peptide repertoires with three reactive cysteine residues, each spaced apart by several random amino acid residues, and we fused the repertoires to the phage gene-3-protein. Conjugation with tris-(bromomethyl) benzene via the reactive cysteines generated repertoires of peptide conjugates with two peptide loops anchored to a mesitylene core. Iterative affinity selections yielded several enzyme inhibitors; after further mutagenesis and selection, we were able to chemically synthesize a lead inhibitor (PK15; K(i) = 1.5 nM) specific to human plasma kallikrein that efficiently interrupted the intrinsic coagulation pathway in human plasma tested ex vivo. This approach offers a powerful means of generating and selecting bicyclic macrocycles (or if cleaved, linear derivatives thereof) as ligands poised at the interface of small-molecule drugs and biologics.