Füllbeck Melanie, Huang Xiaohua, Dumdey Renate, Frommel Cornelius, Dubiel Wolfgang, Preissner Robert
Institute of Biochemistry, Charité, Universitätsmedizin Berlin, Monbijoustr, 2, 10117 Berlin, Germany.
BMC Cancer. 2005 Aug 5;5:97. doi: 10.1186/1471-2407-5-97.
Inhibition of the COP9 signalosome (CSN) associated kinases CK2 and PKD by curcumin causes stabilization of the tumor suppressor p53. It has been shown that curcumin induces tumor cell death and apoptosis. Curcumin and emodin block the CSN-directed c-Jun signaling pathway, which results in diminished c-Jun steady state levels in HeLa cells. The aim of this work was to search for new CSN kinase inhibitors analogue to curcumin and emodin by means of an in silico screening method.
Here we present a novel method to identify efficient inhibitors of CSN-associated kinases. Using curcumin and emodin as lead structures an in silico screening with our in-house database containing more than 10(6) structures was carried out. Thirty-five compounds were identified and further evaluated by the Lipinski's rule-of-five. Two groups of compounds can be clearly discriminated according to their structures: the curcumin-group and the emodin-group. The compounds were evaluated in in vitro kinase assays and in cell culture experiments.
The data revealed 3 compounds of the curcumin-group (e.g. piceatannol) and 4 of the emodin-group (e.g. anthrachinone) as potent inhibitors of CSN-associated kinases. Identified agents increased p53 levels and induced apoptosis in tumor cells as determined by annexin V-FITC binding, DNA fragmentation and caspase activity assays.
Our data demonstrate that the new in silico screening method is highly efficient for identifying potential anti-tumor drugs.
姜黄素对COP9信号体（CSN）相关激酶CK2和PKD的抑制作用可导致肿瘤抑制因子p53稳定。研究表明，姜黄素可诱导肿瘤细胞死亡和凋亡。姜黄素和大黄素可阻断CSN介导的c-Jun信号通路，从而导致HeLa细胞中c-Jun稳态水平降低。本研究旨在通过计算机模拟筛选方法寻找与姜黄素和大黄素类似的新型CSN激酶抑制剂。
本文介绍了一种鉴定CSN相关激酶有效抑制剂的新方法。以姜黄素和大黄素为先导结构，对我们内部包含超过10^6个结构的数据库进行了计算机模拟筛选。鉴定出35种化合物，并根据Lipinski五规则进行进一步评估。根据结构可将两组化合物清晰区分：姜黄素组和大黄素组。对这些化合物进行了体外激酶测定和细胞培养实验。
数据显示，姜黄素组有3种化合物（如白皮杉醇）和大黄素组有4种化合物（如蒽醌）是CSN相关激酶的有效抑制剂。通过膜联蛋白V-FITC结合、DNA片段化和半胱天冬酶活性测定确定，所鉴定的药物可提高p53水平并诱导肿瘤细胞凋亡。
我们的数据表明，新的计算机模拟筛选方法在鉴定潜在抗肿瘤药物方面具有很高的效率。
