Targeting multiple conformations leads to small molecule inhibitors of the uPAR·uPA protein-protein interaction that block cancer cell invasion.
Targeting multiple conformations leads to small molecule inhibitors of the uPAR·uPA protein-protein interaction that block cancer cell invasion.
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DOI：
10.1021/cb200180m
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发表时间：
2011-11-18
影响因子：
4
通讯作者：
Meroueh SO
中科院分区：
文献类型：
--
作者：
Khanna M;Wang F;Jo I;Knabe WE;Wilson SM;Li L;Bum-Erdene K;Li J;W Sledge G;Khanna R;Meroueh SO
尿激酶受体（uPAR）与其结合伙伴（包括细胞表面尿激酶型纤溶酶原激活物（uPA））的相互作用触发一系列蛋白水解和信号事件，促进侵袭和转移。在这里，我们报告了一种小分子（IPR-456）及其衍生物的发现，这种小分子（IPR-456）可以抑制uPAR·uPA蛋白之间的紧密相互作用。IPR-456是通过对从显式溶剂分子动力学模拟中采样的uPAR的多个构象进行虚拟筛选发现的。生化表征表明，该化合物以亚微摩尔亲和力（Kd = 310 nM）与uPAR结合，抑制蛋白-蛋白紧密相互作用，IC50为10 μM。基于显式溶剂分子动力学模拟的自由能计算表明，羧酸段对IPR-456的重要性，这被包括IPR-803在内的几个衍生物的活性所证实。免疫荧光成像显示，IPR-456抑制乳腺癌MDA-MB-231肿瘤细胞uPA与uPAR的结合，IC50为8 μM。这些化合物阻断了MDA-MB-231的细胞侵袭，但IPR-456对MDA-MB-231的迁移几乎没有影响，对粘附也没有影响，这表明uPAR通过其其他结合伙伴介导了这些过程。
Interaction of the urokinase receptor (uPAR) with its binding partners including the urokinase-type plasminogen activator (uPA) at the cell surface triggers a series of proteolytic and signaling events that promote invasion and metastasis. Here, we report the discovery of a small molecule (IPR-456) and its derivatives that inhibit the tight uPAR·uPA protein-protein interaction. IPR-456 was discovered by virtual screening against multiple conformations of uPAR sampled from explicit-solvent molecular dynamics simulations. Biochemical characterization reveal that the compound binds to uPAR with sub-micromolar affinity (Kd = 310 nM) and inhibits the tight protein-protein interaction with an IC50 of 10 μM. Free energy calculations based on explicit-solvent molecular dynamics simulations suggested the importance of a carboxylate moiety on IPR-456, which was confirmed by the activity of several derivatives including IPR-803. Immunofluorescence imaging showed that IPR-456 inhibited uPA binding to uPAR of breast MDA-MB-231 tumor cells with an IC50 of 8 μM. The compounds blocked MDA-MB-231 cell invasion, but IPR-456 showed little effect on MDA-MB-231 migration, and no effect on adhesion, suggesting that uPAR mediates these processes through its other binding partners.