Identification and characterization of inhibitors of cytoplasmic 5′-nucleotidase cN-II issued from virtual screening
Identification and characterization of inhibitors of cytoplasmic 5′-nucleotidase cN-II issued from virtual screening
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DOI：
10.1016/j.bcp.2012.11.024
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发表时间：
2013-02-15
影响因子：
5.8
通讯作者：
Chaloin, Laurent
中科院分区：
文献类型：
--
作者：
Jordheim, Lars Petter;Marton, Zsuzsanna;Chaloin, Laurent
临床和临床前观察表明，5'-核苷酸酶cN-II可能是肿瘤治疗的靶点，无论是本身还是增加细胞毒性核苷类似物的活性。为了确定潜在的cN-II抑制剂，我们对可免费获得的化学数据库进行了硅筛选，用重组cN-II进行了体外酶分析，用截短的cN-II晶体进行了浸泡实验，并对选定的化合物单独或与细胞毒性核苷类似物联合对癌细胞进行了生物学评估。虚拟筛选中排名最高的化合物包括蒽醌衍生物（AdiS），其K-i为2.0 mM。用截断的cN-II晶体进行的浸泡实验可以获得2.9埃的晶体学数据，并表明AdiS与位于cN-II效应位点1的F354/I152之间的相互作用。此外，该化合物在体外对几种癌细胞表现出不同程度的细胞毒性，并在0.5或1.51 μ M克拉瑞滨、0.05 μ M氯法拉滨或30 μ M氟达拉滨孵育的RL细胞中增强了对细胞凋亡的诱导作用。最后，AdiS与克拉宾表现出协同作用，与氯法拉滨表现出可加性。这项研究表明，虚拟筛选是鉴定有效的cN-II抑制剂的有用工具，我们的生物学结果表明，一种先导化合物具有有趣的活性，可以进一步开发为治疗药物。(C) 2012爱思唯尔公司版权所有。
Clinical and preclinical observations have lead to the hypothesis that 5'-nucleotidase cN-II could constitute a therapeutic target in oncology, either per se or to increase the activity of cytotoxic nucleoside analogs.To identify potential cN-II inhibitors, we performed in silico screening of freely available chemical databases, in vitro enzymatic assays with recombinant cN-II, soaking experiments with crystals of truncated cN-II as well as biological evaluation of selected compounds, alone or in combination with cytotoxic nucleoside analogs, on cancer cells.The top ranked compounds from virtual screening included an anthraquinone derivative (AdiS) that were shown to block the enzyme activity with a K-i of 2.0 mM. Soaking experiments performed with crystals of truncated cN-II allowed to obtain crystallographic data at a resolution of 2.9 angstrom and indicating interaction between AdiS and F354/I152 situated in the effector site 1 of cN-II. In addition, this compound exhibited different levels of cytotoxicity in vitro on several cancer cell lines and increased the induction of apoptosis in RL cells incubated with 0.5 or 1.51 mu M cladribine, 0.05 mu M clofarabine or 30 mu M fludarabine. Finally, AdiS showed synergy with cladribine and additivity with clofarabine.This study showed that virtual screening is a useful tool for the identification of potent cN-II inhibitors, and our biological results indicated interesting activity for one lead compound that can be further developed as therapeutics. (C) 2012 Elsevier Inc. All rights reserved.