Benzylidene-indolinones are effective as multi-targeted kinase inhibitor therapeutics against hepatocellular carcinoma.
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.
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Block S4, Singapore 117543, Singapore. Electronic address: phahohk@nus.edu.sg.
Han Kiat
HK
Ho
Singapore OncoGenome Laboratory, Institute of Molecular and Cell Biology, A*STAR, Singapore.
Boon Tin
BT
Chua
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.
Winnie
W
Wong
Singapore OncoGenome Laboratory, Institute of Molecular and Cell Biology, A*STAR, Singapore.
Kah Suan
KS
Lim
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Block S4, Singapore 117543, Singapore.
Vivian
V
Teo
Laboratory of Cancer Genomics, Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore.
Hooi-Tin
HT
Ong
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Block S4, Singapore 117543, Singapore.
Xiao
X
Chen
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Block S4, Singapore 117543, Singapore.
Wei
W
Zhang
Laboratory of Cancer Genomics, Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore.
Kam M
KM
Hui
Department of Pharmacy, National University of Singapore, 18 Science Drive 4, Block S4, Singapore 117543, Singapore.
Mei Lin
ML
Go
Singapore OncoGenome Laboratory, Institute of Molecular and Cell Biology, A*STAR, Singapore; Department of Molecular Biology, Max-Planck-Institute of Biochemistry, Martinsried, Germany.
Axel
A
Ullrich
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D001597:Benzylidene Compounds / Q000737:chemistry / Q000627:therapeutic use*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D007211:Indoles / Q000737:chemistry / Q000627:therapeutic use*; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D000078183:Oxindoles; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000627:therapeutic use*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
D000970:Antineoplastic Agents; D001597:Benzylidene Compounds; D007211:Indoles; D000078183:Oxindoles; D047428:Protein Kinase Inhibitors; C022960:2-oxindole; D011505:Protein-Tyrosine Kinases
Alpha-fetoprotein; Hepatocellular carcinoma; Indolinones; Tyrosine kinase inhibitors; Xenograft