Phosphono Bisbenzguanidines as Irreversible Dipeptidomimetic Inhibitors and Activity-Based Probes of Matriptase-2
Phosphono Bisbenzguanidines as Irreversible Dipeptidomimetic Inhibitors and Activity-Based Probes of Matriptase-2
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DOI：
10.1002/chem.201600206
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发表时间：
2016-06-13
影响因子：
4.3
通讯作者：
Guetschow, Michael
中科院分区：
文献类型：
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作者：
Haeussler, Daniela;Mangold, Martin;Guetschow, Michael
间质蛋白酶-2是一种II型跨膜丝氨酸蛋白酶，在人体铁稳态中起关键作用。抑制matriptase-2被认为是治疗铁超载疾病（例如血色素沉着症和b-地中海贫血）的一种有吸引力的策略。在本研究中，合成路线九二肽模拟灭活剂的开发。鉴定了五种活性化合物（41-45），并在动力学上表征为间质蛋白酶-2的不可逆抑制剂。除了膦酸酯弹头之外，这些二肽还具有两个苯胍部分作为精氨酸模拟物，以通过分别结合S1和S3/S4亚袋来提供对间质蛋白酶-2的亲和力。这种结合模式得到了共价对接分析的有力支持。化合物41-45作为两种非对映异构体的混合物获得，因此分离成单一差向异构体。化合物45 A在N-末端氨基酸处具有S构型，在膦酸酯碳原子处具有R构型，是最有效的间质蛋白酶-2灭活剂，其灭活速率常数为2790 m（-1）s（-1），并且消除了完整细胞表面上的膜结合间质蛋白酶-2的活性。基于膦酰基双苯胍的化学性质，设计并合成了一种插入香豆素标记的荧光探针（51 A）。通过应用51 A作为该酶的第一个基于活性的探针，证明了间质蛋白酶-2的凝胶内荧光检测。
Matriptase-2, a type II transmembrane serine protease, plays a key role in human iron homeostasis. Inhibition of matriptase-2 is considered as an attractive strategy for the treatment of iron-overload diseases, such as hemochromatosis and b-thalassemia. In the present study, synthetic routes to nine dipeptidomimetic inactivators were developed. Five active compounds (41-45) were identified and characterized kinetically as irreversible inhibitors of matriptase-2. In addition to a phosphonate warhead, these dipeptides possess two benzguanidine moieties as arginine mimetics to provide affinity for matriptase-2 by binding to the S1 and S3/S4 subpockets, respectively. This binding mode was strongly supported by covalent docking analysis. Compounds 41-45 were obtained as mixtures of two diastereomers and were therefore separated into the single epimers. Compound 45A, with S configuration at the N-terminal amino acid and R configuration at the phosphonate carbon atom, was the most potent matriptase-2 inactivator with a rate constant of inactivation of 2790 m(-1) s(-1) and abolished the activity of membrane-bound matriptase-2 on the surface of intact cells. Based on the chemotyp of phosphono bisbenzguanidines, the design and synthesis of a fluorescent probe (51A) by insertion of a coumarin label is described. The in-gel fluorescence detection of matriptase-2 was demonstrated by applying 51A as the first activity-based probe for this enzyme.