Engineering inhibitors highly selective for the S1 sites of Ser190 trypsin-like serine protease drug targets
Engineering inhibitors highly selective for the S1 sites of Ser190 trypsin-like serine protease drug targets
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DOI：
10.1016/s1074-5521(01)00084-9
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发表时间：
2001-11-01
影响因子：
--
通讯作者：
Mackman, RL
中科院分区：
文献类型：
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作者：
Katz, BA;Sprengeler, PA;Mackman, RL
背景资料：胰蛋白酶样丝氨酸蛋白酶涉及或牵涉到广泛的疾病，包括充分研究的药物靶标和抗靶标，其可以细分为两大类。在一类中，在S1位点的190位有丝氨酸，如在尿激酶型纤溶酶原激活物（尿激酶或uPA）和因子VIIa中，而在另一类中，在190位有丙氨酸，如在组织型纤溶酶原激活物（tPA）和因子Xa中。O γ（Ser 190）和抑制剂脒之间的Ser 190蛋白酶芳基脒复合物所特有的氢键赋予了这种抑制剂对Ser 190蛋白酶相对于Ala 190对应物的内在偏好。基于Ser 190和Ala 190蛋白酶-芳基脒复合物的S1位点之间的结构差异，我们放大了脒抑制剂对uPA和对tPA的选择性，通过在先导抑制剂支架的脒的邻位引入卤代基团，比较这种卤素取代的和母体抑制剂对一组Ser 190和Ala 190蛋白酶的Ki值，证明卤素类似物对Ser 190蛋白酶的选择性明显高于Ala 190对应物。Ser 190蛋白酶，uPA和胰蛋白酶，以及Ala 190对应物，凝血酶的晶体结构，由一组邻位（卤代，脒基）芳基抑制剂和非卤代亲本的结构基础揭示了精致的选择性，并验证了设计原理。相对于高度相似的tPA抗UPA靶点，非常小的抑制剂实现了对UPA靶点的显著选择性增强。通过在其它相对非选择性支架上单原子取代来靶向。在生理pH下，uPA的总体选择性超过tPA高达980倍。选择性的增加是由于uPA靶标和tPA抗靶标的S1位点共有的单个结合水分子的置换所致，这是因为当结合在Ala 190蛋白酶抗靶标中时，芳基脒抑制剂的氢键键合随之不足。(C)2001爱思唯尔科技有限公司版权所有。
Background: Involved or implicated in a wide spectrum of diseases, trypsin-like serine proteases comprise well studied drug targets and anti-targets that can be subdivided into two major classes. In one class there is a serine at position 190 at the S1 site, as in urokinase type plasminogen activator (urokinase or uPA) and factor VIIa, and in the other there is an alanine at 190, as in tissue type plasminogen activator (tPA) and factor Xa. A hydrogen bond unique to Ser190 protease arylamidine complexes between O gamma (Ser190) and the inhibitor amidine confers an intrinsic preference for such inhibitors toward Ser190 proteases over Ala190 counterparts.Results: Based on the structural differences between the S1 sites of Ser190 and Ala190 protease-arylamidine complexes, we amplified the selectivity of amidine inhibitors toward uPA and against tPA, by factors as high as 220-fold, by incorporating a halo group ortho to the amidine of a lead inhibitor scaffold. Comparison of K-i values of such halo-substituted and parent inhibitors toward a panel of Ser190 and Ala190 proteases demonstrates pronounced selectivity of the halo analogs for Ser190 proteases over Ala190 counterparts. Crystal structures of Ser190 proteases, uPA and trypsin, and of an Ala190 counterpart, thrombin, bound by a set of ortho (halo, amidino) aryl inhibitors and of non-halo parents reveal the structural basis of the exquisite selectivity and validate the design principle.Conclusions: Remarkable selectivity enhancements of exceptionally small inhibitors are achieved toward the uPA target over the highly similar tPA anti-target through a single atom substitution on an otherwise relatively non-selective scaffold. Overall selectivities for uPA over tPA as high as 980-fold at physiological pH were realized. The increase in selectivity results from the displacement of a single bound water molecule common to the S1 site of both the uPA target and the tPA anti-target because of the ensuing deficit in hydrogen bonding of the arylamidine inhibitor when bound in the Ala190 protease anti-target. (C) 2001 Elsevier Science Ltd. All rights reserved.