Ho Han Kiat, Chua Boon Tin, Wong Winnie, Lim Kah Suan, Teo Vivian, Ong Hooi-Tin, Chen Xiao, Zhang Wei, Hui Kam M, Go Mei Lin, Ullrich Axel
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Block S4, Singapore 117543, Singapore.
Singapore OncoGenome Laboratory, Institute of Molecular and Cell Biology, A*STAR, Singapore.
Mol Oncol. 2014 Oct;8(7):1266-77. doi: 10.1016/j.molonc.2014.04.008. Epub 2014 May 2.
Effective pharmacological intervention of advanced hepatocellular carcinoma (HCC) is currently lacking. Despite the use of tyrosine kinase inhibitors (TKIs) for the targeted therapy of several malignancies, no agent has been developed to specifically interfere with the oncogenic tyrosine kinase signaling aberrations found in HCC. Therefore, we adopted an orthogonal biological phenotypic screening approach to uncover candidate compounds: based on a potent cytotoxicity toward HCC-derived cell lines, and minimal toxicity toward normal liver cells. Given the success of indolinone as a chemical scaffold in deriving potent multi-kinase inhibitors (e.g. sunitinib), we screened a group of newly synthesized benzylidene-indolinones. Among the candidates, E/Z 6-Chloro-3-(3-trifluoromethyl-benzyliden)-1,3-dihydroindol-2-one (compound 47) exhibited potent anti-proliferative, anti-migratory, pro-apoptotic properties and good safety profile as compared to known multi-targeted tyrosine kinase inhibitors sunitinib and sorafenib. Additionally, an accompanying suppression of alpha-fetoprotein (AFP) transcription, an HCC tumor marker, implies a favorable selectivity and efficacy on HCC. The in vivo efficacy was demonstrated in an HCC xenograft where 47 was administered once weekly (60 mg/kg) and suppressed tumor burden to the same extent as sorafenib (30 mg/kg daily). A receptor tyrosine kinase (RTK) array study revealed promising inhibition of multiple tyrosine kinases such as IGF-1R, Tyro3 and EphA2 phosphorylation. Gene silencing of these targets ameliorated the cytotoxic potential of 47 on the HuH7 cell line, thereby implicating their contribution to the tumorigenicity of HCC. Hence, 47 exhibits potent anti-cancer effects on HCC cell lines, and is a suitable lead for developing multi-targeted kinase inhibitors of relevance to HCC.
目前，晚期肝细胞癌（HCC）缺乏有效的药物干预措施。尽管酪氨酸激酶抑制剂（TKIs）已用于多种恶性肿瘤的靶向治疗，但尚未开发出专门针对HCC中发现的致癌酪氨酸激酶信号异常的药物。因此，我们采用了一种正交生物表型筛选方法来发现候选化合物：基于对HCC衍生细胞系的强效细胞毒性以及对正常肝细胞的最小毒性。鉴于吲哚酮作为一种化学骨架在开发强效多激酶抑制剂（如舒尼替尼）方面取得了成功，我们筛选了一组新合成的亚苄基吲哚酮。在这些候选化合物中，E/Z 6-氯-3-（3-三氟甲基-亚苄基）-1,3-二氢吲哚-2-酮（化合物47）与已知的多靶点酪氨酸激酶抑制剂舒尼替尼和索拉非尼相比，表现出强效的抗增殖、抗迁移、促凋亡特性以及良好的安全性。此外，伴随的肝癌肿瘤标志物甲胎蛋白（AFP）转录抑制表明对HCC具有良好的选择性和疗效。在HCC异种移植模型中证明了其体内疗效，其中每周一次给予化合物47（60 mg/kg），其抑制肿瘤负荷的程度与索拉非尼（每日30 mg/kg）相同。受体酪氨酸激酶（RTK）阵列研究显示，化合物47有望抑制多种酪氨酸激酶，如IGF-1R、Tyro3和EphA2的磷酸化。对这些靶点进行基因沉默可改善化合物47对HuH7细胞系的细胞毒性潜力，从而表明它们对HCC致瘤性的作用。因此，化合物47对HCC细胞系具有强效抗癌作用，是开发与HCC相关的多靶点激酶抑制剂的合适先导化合物。
