Fragment-based design for the development of N-domain-selective angiotensin-1-converting enzyme inhibitors.
Fragment-based design for the development of N-domain-selective angiotensin-1-converting enzyme inhibitors.
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DOI：
10.1042/cs20130403
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发表时间：
2014-02
期刊：
影响因子：
--
通讯作者：
Sturrock ED
中科院分区：
文献类型：
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作者：
Douglas RG;Sharma RK;Masuyer G;Lubbe L;Zamora I;Acharya KR;Chibale K;Sturrock ED
ACE（血管紧张素1转换酶）是一种锌金属肽酶，在血压调节和电解质稳态中发挥着重要作用。 ACE 由两个同源结构域组成，尽管序列和拓扑结构相似，但在底物加工和抑制剂结合方面显示出差异。选择性抑制 N 结构域（N 选择性）的抑制剂的设计可用于治疗由于 N 结构域特异性底物 Ac-SDKP（N-乙酰基-Ser-Asp-Lys-Pro）积聚而导致的组织损伤和纤维化。使用基于受体的 SHOP（支架跳跃）方法和 N-选择性抑制剂 RXP407，生成了由具有新化学型的修饰 RXP407 主链组成的支架候选列表。这些支架是根据与 N 结构域残基（不同于其 C 结构域对应物）增强的预测相互作用能量来选择的。合成了一种支架并使用荧光 ACE 测定测试了抑制性结合。在 P2 位点掺入四唑部分的分子（化合物 33RE）表现出有效的抑制作用 (Ki=11.21±0.74 nM)，并且对 N 结构域的选择性比对 C 结构域的选择性高 927 倍。与 N 结构域复合的化合物 33RE 的晶体结构揭示了其通过与 His388 的芳族堆积以及与 N 结构域特异性 Tyr369 的羟基的直接氢键结合的模式。这项工作进一步阐明了 N 结构域选择性抑制的分子基础，并有助于设计可用于治疗纤维化疾病的新型 N 选择性 ACE 抑制剂。本文报道了 N 结构域选择性 ACE 抑制剂的开发以及关键活性位点残基与配体相互作用的分子基础。
ACE (angiotensin-1-converting enzyme) is a zinc metallopeptidase that plays a prominent role in blood pressure regulation and electrolyte homeostasis. ACE consists of two homologous domains that despite similarities of sequence and topology display differences in substrate processing and inhibitor binding. The design of inhibitors that selectively inhibit the N-domain (N-selective) could be useful in treating conditions of tissue injury and fibrosis due to build-up of N-domain-specific substrate Ac-SDKP (N-acetyl-Ser–Asp–Lys–Pro). Using a receptor-based SHOP (scaffold hopping) approach with N-selective inhibitor RXP407, a shortlist of scaffolds that consisted of modified RXP407 backbones with novel chemotypes was generated. These scaffolds were selected on the basis of enhanced predicted interaction energies with N-domain residues that differed from their C-domain counterparts. One scaffold was synthesized and inhibitory binding tested using a fluorogenic ACE assay. A molecule incorporating a tetrazole moiety in the P2 position (compound 33RE) displayed potent inhibition (Ki=11.21±0.74 nM) and was 927-fold more selective for the N-domain than the C-domain. A crystal structure of compound 33RE in complex with the N-domain revealed its mode of binding through aromatic stacking with His388 and a direct hydrogen bond with the hydroxy group of the N-domain specific Tyr369. This work further elucidates the molecular basis for N-domainselective inhibition and assists in the design of novel N-selective ACE inhibitors that could be employed in treatment of fibrosis disorders. The present paper reports the development of an N-domain selective ACE inhibitor and the molecular basis for the interaction of key active site residues with the ligand.