Development and Characterization of a Fluorescent Probe for GLS1 and the Application for High-Throughput Screening of Allosteric Inhibitors.
Development and Characterization of a Fluorescent Probe for GLS1 and the Application for High-Throughput Screening of Allosteric Inhibitors.
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DOI：
10.1021/acs.jmedchem.9b01035
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发表时间：
2019-10
影响因子：
7.3
通讯作者：
Xi Xu;Zijian Kuang;Jie Han;Yingfen Meng;Lei Li;Hongyu Luan;Pengfei Xu;Jubo Wang;C. Luo;H. Ding;Zhiyu Li;Jinlei Bian
中科院分区：
文献类型：
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作者：
Xi Xu;Zijian Kuang;Jie Han;Yingfen Meng;Lei Li;Hongyu Luan;Pengfei Xu;Jubo Wang;C. Luo;H. Ding;Zhiyu Li;Jinlei Bian
谷氨酰胺酶（Glutaminase，GLS 1）是一种在细胞生长和增殖中起重要作用的肿瘤能量代谢蛋白。由于其主要参与恶性肿瘤的发生，迫切需要小分子GLS 1抑制剂来评估其治疗潜力并探索其潜在的生物学功能。最近的研究表明，靶向变构结合位点代表了一种有前途的策略，用于识别有效的和选择性的GLS 1抑制剂。在此，我们提出了两个荧光探针的合成靶向的变构结合位点的GLS 1和它们的用途作为机械工具，在多个适用的检测平台。基于荧光偏振（FP）的结合测定能够从我们通过点击化学方法获得的内部化合物库中简单、快速和可靠地筛选变构抑制剂。实验结果表明，C147（命名为CPU-L1）比对照化合物CB 839具有更强的抑制活性和更好的溶解性，有望作为新型GLS 1抑制剂进一步优化。
Glutaminase (GLS1) is a cancer energy metabolism protein which plays a predominant role in cell growth and proliferation. Due to its major involvement in malignant tumor, small-molecule GLS1 inhibitors are urgently needed to assess its therapeutic potential and for probing their underlying biology function. Recent studies showed that targeting the allosteric binding site represented a promising strategy for identifying potent and selective GLS1 inhibitors. Herein, we present the synthesis of two fluorescent probes targeting the allosteric binding site of GLS1 and their usage as mechanistic tools in multiple applicable assay platform. The fluorescence polarization (FP)-based binding assay enables easy, fast and reliable screen of allosteric inhibitors from our in-house compound library obtained through click chemistry method. The obtained compound C147 (named as CPU-L1) has been proved to be more potent and with greater solubility than the control compound CB839, which could serve as promising leads for further optimization as novel GLS1 inhibitors.