GLI1 Inhibitors Identified by Target Protein Oriented Natural Products Isolation (TPO-NAPI) with Hedgehog Inhibition
GLI1 Inhibitors Identified by Target Protein Oriented Natural Products Isolation (TPO-NAPI) with Hedgehog Inhibition
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DOI：
10.1021/acschembio.8b00492
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发表时间：
2018-09-01
影响因子：
4
通讯作者：
Ishibashi, Masami
中科院分区：
文献类型：
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作者：
Arai, Midori A.;Ochi, Fumie;Ishibashi, Masami
本文介绍了一种针对Hedgehog抑制剂和直接GLI1抑制剂5‘-O-甲基-3-羟基flemingin A(3)的目标蛋白导向天然产物分离(TPO-NAPI)方法的发展，该抑制剂可抑制Hedgehog(HH)信号转导并降低肿瘤干细胞的特性。利用GLI1蛋白固定化小球开发的TPO-NAPI方法，快速获得了8个直接与GLI1结合的天然产物(包括3个新描述的产物)。5‘-O-甲基-3-羟基荧光素A(3)抑制HH信号转导(IC50 7.3µM)，导致HH靶蛋白BCL2、ptch1和BMI1的产生减少。5‘-O-甲基-3-羟基荧光素A(3)对HH相关癌细胞具有细胞毒作用。CD实验表明，5‘-0-甲基-3-羟基荧光素A(3)直接与GM结合(K-d=7.7mM)。此外，5‘-0-甲基-3-羟基荧光素A(3)抑制了HuH7的肿瘤干细胞特征，如球体的形成和肿瘤干细胞标记物EpCAM的产生。这些结果表明，HH抑制剂可以有效地抑制肿瘤干细胞的活性。
This report describes the development of a target-protein-oriented natural-products-isolation (TPO-NAPI) method for Hedgehog inhibitors and the direct GLI1 inhibitor, 5'-O-methyl-3-hydroxyflemingin A (3), which inhibited hedgehog (Hh) signal transduction and diminished characteristics of cancer stem cells. Eight natural products (including three newly described products) that directly bind to GLI1 were rapidly obtained via the TPO-NAPI method developed using GLI1 protein-immobilized beads. 5'-O-Methyl-3-hydroxyflemingin A (3) inhibited Hh signaling (IC50 7.3 mu M), leading to decreasing production of the Hh target proteins BCL2, PTCH1, and BMI1. 5'-O-Methyl-3-hydroxyflemingin A (3) was cytotoxic to Hh-related cancer cells. CD experiments revealed that 5'-0-methyl-3-hydroxyflemingin A (3) directly bound GM (K-d = 7.7 mu M). Moreover, 5'-0-methyl-3-hydroxyflemingin A (3) diminished cancer stem cell characters of Huh7 such as sphere formation and production of the cancer stem cell marker EpCAM. These results suggest that Hh inhibitors can efficiently suppress the activity of cancer stem cells.