Zeslawska Ewa, Jacob Uwe, Schweinitz Andrea, Coombs Gary, Bode Wolfram, Madison Edwin
Pedagogical University, Department of Chemistry, Podchorazych 2, 30-084 Cracow, Poland.
J Mol Biol. 2003 Apr 18;328(1):109-18. doi: 10.1016/s0022-2836(03)00267-5.
Urokinase type plasminogen activator (uPA), a trypsin-like serine proteinase, plays an important role in normal tissue re-modelling, cell adhesion, and cell motility. In addition, studies utilizing normal animals and potent, selective uPA inhibitors or genetically modified mice that lack functional uPA genes have demonstrated that uPA can significantly enhance tumor initiation, growth, progression and metastasis, strongly suggesting that this enzyme may be a promising anti-cancer target. We have investigated the structure-activity relationship (SAR) of peptidomimetic inhibitors of uPA and solved high resolution X-ray structures of key, lead small molecule inhibitors (e.g. phenethylsulfonamidino(P4)-D-seryl(P3)-L-alanyl(P2)-L-argininal(P1) and derivatives thereof) in complex with the uPA proteinase domain. These potent inhibitors are highly selective for uPA. The non-natural D-seryl residue present at the P3 position in these inhibitors contributes substantially to both potency and selectivity because, due to its D-configuration, its side-chain binds in the S4 pocket to interact with the uPA unique residues Leu97b and His99. Additional potency and selectivity can be achieved by optimizing the inhibitor P4 residue to bind a pocket, known as S1sub or S1beta, that is adjacent to the primary specificity pocket of uPA.
尿激酶型纤溶酶原激活剂(uPA)是一种胰蛋白酶样丝氨酸蛋白酶,在正常组织重塑、细胞黏附和细胞运动中发挥重要作用。此外,利用正常动物以及强效、选择性uPA抑制剂或缺乏功能性uPA基因的基因工程小鼠进行的研究表明,uPA可显著增强肿瘤的起始、生长、进展和转移,强烈提示该酶可能是一个有前景的抗癌靶点。我们研究了uPA拟肽类抑制剂的构效关系(SAR),并解析了关键先导小分子抑制剂(如苯乙基磺脒基(P4)-D-丝氨酰(P3)-L-丙氨酰(P2)-L-精氨醛(P1)及其衍生物)与uPA蛋白酶结构域复合物的高分辨率X射线结构。这些强效抑制剂对uPA具有高度选择性。这些抑制剂P3位存在的非天然D-丝氨酰残基对效力和选择性均有显著贡献,因为由于其D-构型,其侧链结合在S4口袋中,与uPA独特的残基Leu97b和His99相互作用。通过优化抑制剂的P4残基以结合一个与uPA主要特异性口袋相邻的口袋(称为S1sub或S1beta),可以实现额外的效力和选择性。