Abstract
Interleukin 6 and the signal transducer and activator of transcription (STAT) 3 proteins have important roles in cancer cell survival and proliferation. Recent studies demonstrate that abnormal STAT3 activation promotes tumor growth and supports survival of many human cancers, and thus, this protein or the pathway responsible for its activation is a potential target for the new anticancer therapy. STAT3 is a DNA binding transcription factor, and therefore, its function depends on nuclear translocation. To discover inhibitors of the STAT3 pathway, we designed a cell-based screening assay capable of identifying small molecules that inhibit nuclear translocation. Among the 2000-compound National Cancer Institute Diversity set, we identified 8-benzyl-4-oxo-8-azabicyclo[3.2.1]oct-2-ene-6,7-dicarboxylic acid (SD-1008) as a micromolar inhibitor of interleukin-6 or oncostatin-induced STAT3 nuclear translocation. In addition, SD-1008 inhibits tyrosyl phosphorylation of STAT3, Janus kinase 2 (JAK2), and Src. SD-1008 also reduces STAT3-dependent luciferase activity. Biochemical studies with recombinant JAK2 proteins demonstrate that high concentrations of SD-1008 directly inhibit JAK2 kinase autophosphorylation. Exposure of various cell lines to SD-1008 decreases levels of the STAT3-dependent proteins, Bcl-XL and survivin, inducing apoptosis. SD-1008 also enhances apoptosis induced by paclitaxel in ovarian cancer cells. These results demonstrate that SD-1008 directly blocks the JAK-STAT3 signaling pathway in human cancer cells that express constitutively active Stat and add to the growing literature that identifies this pathway as a viable target for drug development. Finally, SD-1008 may be a suitable prototype for further chemical modification and exploration as a therapeutic agent.
Footnotes
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This project was supported by the Ovarian Cancer Specialized Program in Research Excellence at the Dana-Farber/Harvard Cancer Center, grant 1P50-CA105009 from the Ovarian Cancer Education and Awareness Network, and support through the Lana Vento Foundation. Z.D. is supported, in part, through a grant from the Marsha Rivkin Foundation. M.V.S. is supported through a Mid Career Development Award, 1K24-CA109416-01A1. J.B. is supported by grants from the Multiple Myeloma Research Foundation and the Clinical Investigator Training Program: Harvard-MIT Health Sciences and Technology–Beth Israel Deaconess Medical Center, in collaboration with Pfizer Inc. and Merck and Company, Inc.
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Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
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doi:10.1124/mol.107.038117.
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ABBREVIATIONS: STAT3, signal transducer and activator of transcription 3; IL-6, interleukin 6; JAK, Janus-activated kinase; NCI, National Cancer Institute; SD-1008, 8-benzyl-4-oxo-8-azabicyclo[3.2.1]oct-2-ene-6,7-dicarboxylic acid; DMSO, dimethyl sulfoxide; EGFP, enhanced green fluorescent protein; PARP, poly(ADP-ribose) polymerase; ELISA, enzyme-linked immunosorbent assay; AG490, α-cyano-(3,4-dihydroxy)-N-benzylcinnamide; SD-1029, 9-(3,4-dichlorophenyl)-2,7-bis(dimethylaminomethyl)-3,4,5,6,7,9-hexahydro-2H-xanthene-1,8-dione.
- Received May 14, 2007.
- Accepted August 3, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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