In vitro selection of highly modified cyclic peptides that act as tight binding inhibitors.
In vitro selection of highly modified cyclic peptides that act as tight binding inhibitors.
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DOI：
10.1021/ja301017y
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发表时间：
2012-06-27
影响因子：
15
通讯作者：
Szostak, Jack W.
中科院分区：
文献类型：
--
作者：
Schlippe, Yollete V. Guillen;Hartman, Matthew C. T.;Josephson, Kristopher;Szostak, Jack W.
人们迫切需要发现新的治疗分子，这些分子将生物药物的高特异性和亲和力与小分子的生物利用度和低成本相结合。小的天然产物样肽在弥合这一差距方面有很大的希望;然而，获得这些化合物的库一直是一个限制。由于核糖体肽可以进行体外选择技术，因此高度修饰的大环肽的极大文库（>1013）的产生可以为新的有用的生物活性分子的产生和选择提供强有力的替代。此外，将许多非蛋白质氨基酸掺入核糖体肽中并与大环化结合，应增强这些文库的药物样特征。在这里，我们表明，mRNA展示，一种技术，允许在体外选择的肽，可以适用于大环肽，含有大多数非天然氨基酸的演变。我们描述了两个这样的非天然的环状肽，结合蛋白酶凝血酶与低纳摩尔亲和力的分离和表征，我们表明，在这些肽中的非天然残基是必不可少的观察到的高亲和力结合。我们证明，所选的肽是紧密结合的凝血酶抑制剂，Kiapp值在低纳摩尔范围内。在实验室中进化高度修饰的大环肽的能力是迈向容易产生有用的分子试剂和治疗性先导分子的第一个关键步骤，所述分子试剂和治疗性先导分子将生物制剂的有利特征与小分子药物的有利特征联合收割机结合。
There is a great demand for the discovery of new therapeutic molecules that combine the high specificity and affinity of biologic drugs with the bioavailability and lower cost of small molecules. Small, natural-product-like peptides hold great promise in bridging this gap; however, access to libraries of these compounds has been a limitation. Since ribosomal peptides may be subjected to in vitro selection techniques, the generation of extremely large libraries (>1013) of highly modified macrocyclic peptides may provide a powerful alternative for the generation and selection of new useful bioactive molecules. Moreover, the incorporation of many non-proteinogenic amino acids into ribosomal peptides in conjunction with macrocyclization should enhance the drug-like features of these libraries. Here we show that mRNA-display, a technique that allows the in vitro selection of peptides, can be applied to the evolution of macrocyclic peptides that contain a majority of unnatural amino acids. We describe the isolation and characterization of two such unnatural cyclic peptides that bind the protease thrombin with low nanomolar affinity, and we show that the unnatural residues in these peptides are essential for the observed high-affinity binding. We demonstrate that the selected peptides are tight-binding inhibitors of thrombin, with Kiapp values in the low nanomolar range. The ability to evolve highly modified macrocyclic peptides in the laboratory is the first crucial step toward the facile generation of useful molecular reagents and therapeutic lead molecules that combine the advantageous features of biologics with those of small-molecule drugs.
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影响因子：
14.8
作者：
Heinis, Christian;Rutherford, Trevor;Winter, Greg
通讯作者：
Winter, Greg
DOI：
10.1073/pnas.0509219103
发表时间：
2006-03-21
影响因子：
11.1
作者：
Hartman, MCT;Josephson, K;Szostak, JW
通讯作者：
Szostak, JW
影响因子：
14.9
作者：
Kurz, M;Gu, K;Lohse, P A
通讯作者：
Lohse, P A
影响因子：
3.7
作者：
Hartman, Matthew C. T.;Josephson, Kristopher;Szostak, Jack W.
通讯作者：
Szostak, Jack W.
影响因子：
5.2
作者：
Dewkar, Gajanan K.;Carneiro, Pedro B.;Hartman, Matthew C. T.
通讯作者：
Hartman, Matthew C. T.