Inhibition of thermolysin and neutral endopeptidase 24.11 by a novel glutaramide derivative: X-ray structure determination of the thermolysin-inhibitor complex.
Inhibition of thermolysin and neutral endopeptidase 24.11 by a novel glutaramide derivative: X-ray structure determination of the thermolysin-inhibitor complex.
复制标题
新型戊二酰胺衍生物对嗜热菌蛋白酶和中性内肽酶 24.11 的抑制：嗜热菌蛋白酶抑制剂复合物的 X 射线结构测定。
DOI：
10.1021/bi00167a007
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
James,K
中科院分区：
文献类型：
--
作者：
Holland,DR;Barclay,PL;Danilewicz,JC;Matthews,BW;James,K
1993年10月11日 * 摘要：嗜热菌蛋白酶抑制剂复合物的X射线结构的测定已被证明有助于帮助我们理解相关的、生理上重要的哺乳动物锌肽酶（包括中性内肽酶EC 3.4）的抑制剂的结合模式。24.11和血管紧张素转化酶。本文描述了7V-{1-（2（R，S ′）-羧基-4-苯基丁基）-环戊基羰基}-（S ′）-色氨酸（CCT）与结晶嗜热菌蛋白酶的结合方式。CCT是坎多沙曲拉（中性内肽酶24.11的有效抑制剂）和5-茚满酯前药坎多沙曲（正在临床评估中，作为充血性心力衰竭的潜在治疗方法）的类似物。CCT在几个重要方面不同于先前研究的N-羧烷基二肽CLT [A-（S ′）-（1-羧基-3-苯基丙基）-（S ′）-亮氨酰-（S ′）-色氨酸]。它具有高度受限的偕-环戊基P/取代基，并且缺乏CLT的特征亚氨基氮取代基。结构测定表明，尽管构象的偕环戊基取代基的影响，CCT结合在嗜热菌蛋白酶的活性位点内以类似的方式CLT。虽然CLT和嗜热菌蛋白酶的亚氨基氮之间的特征性氢键在CCT中不存在，但两种抑制剂对酶的亲和力几乎相同。这些结果说明了不仅要考虑在酶-配体复合物中形成的那些氢键，而且要考虑在复合物形成时由于酶和配体的去溶剂化而可能失去的其他氢键的重要性。此外，为了实现受体相互作用，对配体的整体构象要求可能是至关重要的。锌依赖性肽酶在许多生物活性肽的生物合成和代谢中起关键作用。因此，这些酶的抑制剂作为治疗剂具有相当大的潜力（Powers和哈珀，1986; Rich，1990）。在血管紧张素转化酶（ACE）抑制剂的情况下，这种潜力已经得到实现，其已经证明在治疗高血压和充血性心力衰竭中的临床益处（Patchett & Cordes，1985;赖亚等人，1990年）。最近，中性内肽酶EC 3.4的抑制剂。24.11（内肽酶24.11）（Sybertz，1990; Roques等人，1990）和基质金属蛋白酶（Wahl等人，1990）分别作为充血性心力衰竭和类风湿性关节炎的治疗方法。我们对这些生理上重要的哺乳动物肽酶的水解机制及其与抑制剂的相互作用的理解，已经通过相关的细菌内肽酶嗜热菌蛋白酶（TLN）的研究得到了很大的帮助，其X射线晶体结构已经在其天然状态（Holmes &马修斯，1982）和在其非天然状态下（TLN）被报道。
Revised Manuscript Received October 11, 1993* abstract: Determination of the X-ray structure of thermolysin-inhibitor complexes has proven useful in aiding our understanding of the mode of binding of inhibitors of related, physiologically important, mammalian zinc peptidases including neutral endopeptidase EC 3.4. 24.11 and angiotensin-converting enzyme. Here we describe the mode of binding to crystalline thermolysin of 7V-{1-(2 (R, S')-carboxy-4-phenylbutyl)-cyclopentylcarbonyl}-(S')-tryptophan (CCT). CCT is an analogue of both candoxatrilat, a potent inhibitor of neutral endopeptidase 24.11, and of the 5-indanyl ester prodrug candoxatril, which is under clinical evaluation as a potential therapy for congestive heartfailure. CCT differs from the previously studied 7V-carboxyalkyl dipeptide CLT [A-(S’)-(l-carboxy-3-phenylpropyl)-(S,)-leucyl-(S')-tryptophan] in several important respects. It has a highly constrained gem-cyclopentyl P/substituent and lacks the characteristic imino nitrogen substituent of CLT. The structure determination shows that, notwithstanding the conformational influence of the gem-cyclopentyl substituent, CCT binds within the active site of thermolysin in a similar manner to CLT. Although the characteristic hydrogen bond between the imino nitrogen of CLT and thermolysin is absent in CCT, the affinities of the two inhibitors for the enzyme are virtually identical. These results illustrate the importance of considering not only those hydrogen bonds that are formed in an enzyme-ligand complex but also the other hydrogen bonds that may be lost due to desolvation of the enzyme and ligand on formation of thecomplex. In addition, the overall conformational demands placed upon a ligand in order to achieve receptor interaction may be critically important.Zinc-dependent peptidases play a key role in the biosynthesis and metabolism of a number of bioactive peptides. Conse-quently, inhibitors of these enzymes have considerable potential as therapeutic agents (Powers & Harper, 1986; Rich, 1990). This potential has been fulfilled inthe case of angiotensinconverting enzyme (ACE) inhibitors, which have demonstrated clinical benefit in the treatment of hypertension and congestive heart failure (Patchett & Cordes, 1985; Raia et al., 1990). More recently, inhibitors of neutral endopeptidase EC 3.4. 24.11 (endopeptidase 24.11)(Sybertz, 1990; Roques et al., 1990) and matrix metalloproteases (Wahl et al., 1990) are reported to be under investigation as therapies for congestive heartfailure and rheumatoid arthritis, respectively. Our understanding of the hydrolytic mechanism of these physiologically important mammalian peptidases, and their interaction with inhibitors, has been aided greatly by study of the related bacterial endopeptidase thermolysin (TLN), the X-ray crystal structure of which has been reported, both in its native state (Holmes & Matthews, 1982) and in