Fragment-Based Protein-Protein Interaction Antagonists of a Viral Dimeric Protease.
Fragment-Based Protein-Protein Interaction Antagonists of a Viral Dimeric Protease.
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DOI：
10.1002/cmdc.201500526
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发表时间：
2016-04-19
期刊：
影响因子：
3.4
通讯作者：
Craik CS
中科院分区：
文献类型：
--
作者：
Gable JE;Lee GM;Acker TM;Hulce KR;Gonzalez ER;Schweigler P;Melkko S;Farady CJ;Craik CS
基于片段的药物发现已经显示出作为一种挑战目标的方法的前景，例如蛋白质-蛋白质界面。我们开发并应用了一种基于活性的片段筛选，以对抗二聚体卡波西肉瘤相关疱疹病毒蛋白(KSHV Pr)，使用优化的荧光底物。剂量反应测定作为确认筛选，核磁共振波谱用于定位与KSHV Pr结合的片段抑制物。动力学分析表明，几种初始HIT也抑制人巨细胞病毒蛋白水解酶(HCMV Pr)。核磁共振波谱也证实了这些HITS与HCMV Pr的结合。尽管使用了靶标不可知的片段文库，但超过80%的确认命中干扰了二聚反应，并与先前报道的KSHV Pr二聚体界面上的口袋结合，而不是与活性部位结合。使用商业上可获得的类似物进一步研究了一类碎片，即氨基噻唑支架。这些化合物的抑制率提高了100倍以上。这项研究说明了基于片段的筛选这些具有挑战性的酶靶点的能力，并提供了一个在蛋白质-蛋白质界面上的口袋潜在的可药性的例子。
Fragment-based drug discovery has shown promise as an approach for challenging targets such as protein-protein interfaces. We developed and applied an activity-based fragment screen against dimeric Kaposi’s sarcoma-associated herpesvirus protease (KSHV Pr) using an optimized fluorogenic substrate. Dose response determination was performed as a confirmation screen and NMR spectroscopy was used to map fragment inhibitor binding to KSHV Pr. Kinetic assays demonstrated that several initial hits also inhibit human cytomegalovirus protease (HCMV Pr). Binding of these hits to HCMV Pr was also confirmed via NMR spectroscopy. Despite the use of a target-agnostic fragment library, more than 80% of confirmed hits disrupted dimerization and bound to a previously reported pocket at the dimer interface of KSHV Pr, not to the active site. One class of fragments, an aminothiazole scaffold, was further explored using commercially available analogs. These compounds demonstrated greater than 100-fold improvement of inhibition. This study illustrates the power of fragment-based screening for these challenging enzymatic targets and provides an example of the potential druggability of pockets at protein-protein interfaces.