Identification of novel binding interactions in the development of potent, selective 2-naphthamidine inhibitors of urokinase.: Synthesis, structural analysis, and SAR of N-phenyl amide 6-substitution
Identification of novel binding interactions in the development of potent, selective 2-naphthamidine inhibitors of urokinase.: Synthesis, structural analysis, and SAR of N-phenyl amide 6-substitution
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DOI：
10.1021/jm0300072
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发表时间：
2004-01-15
影响因子：
7.3
通讯作者：
Giranda, VL
中科院分区：
文献类型：
--
作者：
Wendt, MD;Rockway, TW;Giranda, VL
描述了丝氨酸蛋白酶尿激酶纤溶酶原激活剂（uPA或尿激酶）的6-取代2-萘脒抑制剂的制备和生物活性评估。基于综合考虑和取代基向量建模，选择 2-萘脒作为起点。发现 6 位的苯酰胺可以改善结合；用其他双原子连接体替换酰胺被证明是无效的。苯基本身位于S1'亚位附近；苯基基团的取代进入S1'和其他远距离结合区域。通过环取代定义和探索了三个新的相互作用点。使用 4-烷基氨基与 Asp60A 羧酸盐形成暴露于溶剂的盐桥，从而提高对 K-i = 40 nM 的亲和力。抑制剂还访问两个疏水区域。一种相互作用的特征是紧密的疏水配合，该配合由主要由 His57 和 His99 定义的小凹坑形成；较弱、特异性较低的相互作用涉及烷基进入 Va141 和 Cys42-Cys58 二硫化物附近的广泛质侧蛋白结合区域，取代水分子并导致活性小幅增加。许多抑制剂访问了这三个区域中的两个。亲和力范围低至 Ki = 6 nM，许多化合物的 K-i < 100 nM，而与相关丝氨酸蛋白酶组的五个成员中的四个相比，获得了中等至优异的选择性。此外，通过与 Asp60A 相互作用产生了一些针对胰蛋白酶的选择性。许多这些化合物的 X 射线结构被用来为我们的抑制剂设计提供信息，并增加我们对关键相互作用的理解。结合我们对 8 取代模式的探索，我们已经确定了 uPA 抑制剂的许多新颖的结合相互作用。
The preparation and assessment of biological activity of 6-substituted 2-naphthamidine inhibitors of the serine protease urokinase plasminogen activator (uPA, or urokinase) is described. 2-Naphthamidine was chosen as a starting point based on synthetic considerations and on modeling of substituent vectors. Phenyl amides at the 6-position were found to improve binding; replacement of the amide with other two-atom linkers proved ineffective. The phenyl group itself is situated near the S1' subsite; substitutions off of the phenyl group accessed S1' and other distant binding regions. Three new points of interaction were defined and explored through ring substitution. A solvent-exposed salt bridge with the Asp60A carboxylate was formed using a 4-alkylamino group, improving affinity to K-i = 40 nM. Inhibitors also accessed two hydrophobic regions. One interaction is characterized by a tight hydrophobic fit made with a small dimple largely defined by His57 and His99; a weaker, less specific interaction involves alkyl groups reaching into the broad prime-side protein binding region near Va141 and the Cys42-Cys58 disulfide, displacing water molecules and leading to small gains in activity. Many inhibitors accessed two of these three regions. Affinities range as low as Ki = 6 nM, and many compounds had K-i < 100 nM, while moderate to excellent selectivity was gained versus four of five members of a panel of relevant serine proteases. Also, some selectivity against trypsin was generated via the interaction with Asp60A. X-ray structures of many of these compounds were used to inform our inhibitor design and to increase our understanding of key interactions. In combination with our exploration of 8-substitution patterns, we have identified a number of novel binding interactions for uPA inhibitors.