Metallopeptide-promoted inactivation of angiotensin-converting enzyme and endothelin-converting enzyme 1: toward dual-action therapeutics
Metallopeptide-promoted inactivation of angiotensin-converting enzyme and endothelin-converting enzyme 1: toward dual-action therapeutics
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DOI：
10.1007/s00775-006-0145-2
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发表时间：
2006-10-01
影响因子：
3
通讯作者：
Cowan, J. A.
中科院分区：
文献类型：
--
作者：
Gokhale, Nikhil H.;Cowan, J. A.
基于氨基末端铜/镍（ATCUN）结合基序的一系列金属肽已被评价为兔和人血管紧张素转换酶（hACE）以及人内皮素转换酶1（hECE-1）的经典抑制剂和催化灭活剂。钴络合物[KGHK-Co（NH3）（2）]（2+），其中KGHK是赖氨酰甘氨酰-组氨酰赖氨酸，显示出与[KGHK-Cu]（+）相似的K-I和IC 50值，尽管电荷增加，因此电荷的影响被轴向配位的氨胺配体的空间影响抵消，或者结合由氨基酸侧链的贡献主导，特别是模拟赖诺普利所观察到的结合模式的C-末端赖氨酸。此外，观察到的[KGHK-Co（NH3）（2）]（2+）的抑制作用与Co2+（aq）对hACE的激活作用形成对比，反映了在两个活性位点中的每一个处用钴离子替换催化性锌辅因子后对酶活性的刺激。定量分析的剂量依赖性的刺激活性的Co 2+（水溶液）产生明显的亲和力为1.3 +/- 0.2和56 +/- 8 μ M的两个网站在饱和的Zn 2+（10 μ M）的存在下。先前已表征了亚饱和浓度下[KGHK-Cu]（+）对hACE的催化灭活作用，k（obs）= 2.9 +/- 0.5 x 10（-2）min（-1）。在类似的条件下，发现相同的络合物催化hECE-1，k（obs）= 2.12 +/- 0.16 x 10（-2）min（-1），证明了针对心血管疾病中两个关键药物靶标的双重作用活性的潜力。药物靶点的不可逆失活代表了一种新的药物作用机制，它补充了现有的经典抑制剂策略，这些策略是当前药物发现工作的基础。
A series of metallopeptides based on the amino terminal copper/nickel (ATCUN) binding motif have been evaluated as classical inhibitors and catalytic inactivators of both rabbit and human angiotensin-converting enzyme (hACE), and human endothelin-converting enzyme 1 (hECE-1). The cobalt complex [KGHK - Co(NH3)(2)] (2+), where KGHK is lysylglycyl-histidyllysine, displayed similar K-I and IC50 values to those found for [KGHK-Cu](+), in spite of the enhanced charge, and so either the influence of charge is offset by the steric influence of the axially coordinated ammine ligands, or binding is dominated by contributions from the amino acid side chains, especially the C-terminal lysine that mimics the binding pattern observed for lisinopril. Moreover, the inhibition observed for [KGHK-Co(NH3)(2)](2+) contrasts with the activation of hACE by Co2+(aq), reflecting the stimulation of enzyme activity following replacement of the catalytic zinc cofactor by cobalt ion at each of the two active sites. Quantitative analysis of the dose-dependent stimulation of activity by Co2+(aq) yielded apparent affinities of 1.3 +/- 0.2 and 56 +/- 8 mu M for the two sites in the presence of saturating Zn2+ (10 mu M). Catalytic inactivation of hACE by [KGHK-Cu](+) at subsaturating concentrations had previously been characterized, with k(obs) = 2.9 +/- 0.5 x 10(-2) min(-1). Under similar conditions, the same complex is found to catalytically inactivate hECE-1, with k(obs) = 2.12 +/- 0.16 x 10(-2) min(-1), demonstrating the potential for dual-action activity against two key drug targets in cardiovascular disease. Irreversible inactivation of a drug target represents a novel mechanism of drug action that complements existing classical inhibitor strategies that underlie current drug discovery efforts.