Peptide Macrocycle Inhibitor of Coagulation Factor XII with Subnanomolar Affinity and High Target Selectivity
Peptide Macrocycle Inhibitor of Coagulation Factor XII with Subnanomolar Affinity and High Target Selectivity
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DOI：
10.1021/acs.jmedchem.6b01548
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发表时间：
2017-02-09
影响因子：
7.3
通讯作者：
Heinis, Christian
中科院分区：
文献类型：
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作者：
Middendorp, Simon J.;Wilbs, Jonas;Heinis, Christian
因子XII（FXII）是近年来作为治疗或预防病理性血栓形成、抑制体外循环中的接触活化以及治疗肿胀病症遗传性血管性水肿的潜在靶标而出现的血浆蛋白酶。虽然开发了几种对活化的FXII（FXIIa）具有高亲和力的基于蛋白质的抑制剂，但小分子抑制剂的产生一直具有挑战性。在这项工作中，我们通过优化最近通过噬菌体展示进化的肽大环化合物（Ki = 0.84 +/- 0.03 nM）产生了有效的和选择性的FXIIa抑制剂。在苯丙氨酸的对位引入的氟原子使结合亲和力提高了10倍之多。此外，我们通过用去甲精氨酸取代N-末端精氨酸来提高蛋白水解稳定性。所得抑制剂结合了高抑制亲和力和选择性以及在血浆中的良好稳定性（Ki = 1.63 +/- 0.18 nM，> 27000倍选择性，t（1/2）（血浆）=16 +/- 4 h）。该抑制剂有效地阻断了人血液中内源性凝血途径的激活。
Factor XII (FXII) is a plasma protease that has emerged in recent years as a potential target to treat or prevent pathological thrombosis, to inhibit contact activation in extracorporeal circulation, and to treat the swelling disorder hereditary angioedema. While several protein based inhibitors with high affinity for activated FXII (FXIIa) were developed, the generation of small molecule inhibitors has been challenging. In this work, we have generated a potent and selective FXIIa inhibitor by optimizing a peptide macrocycle that was recently evolved by phage display (K-i = 0.84 +/- 0.03 nM). A fluorine atom introduced in the para-position of phenylalanine enhanced the binding affinity as much as 10-fold. Furthermore, we improved the proteolytic stability by substituting the N-terminal arginine by norarginine. The resulting inhibitor combines high inhibitory affinity and selectivity with a good stability in plasma (K-i = 1.63 +/- 0.18 nM, >27 000-fold selectivity, t(1/2)(plasma) =16 +/- 4 h). The inhibitor efficiently blocked activation of the intrinsic coagulation pathway in human blood ex vivo.