Abstract
Aim:
To investigate the molecular targets of DCLAK11, a novel compound discovered from a series of substituted pyridin-3-amine derivatives, and to characterize its anti-tumor properties in vitro.
Methods:
Kinase inhibition was measured by an ELISA assay. Cell viability was assessed with an SRB or a CCK8 assay. The alterations induced by kinase signaling proteins in cancer cells were detected by Western blot. Apoptosis was determined by an Annexin V-PI assay. The following assays were used to evaluate the impact on angiogenesis: wound-healing, Transwell, tube formation and microvessel outgrowth from rat aortic rings.
Results:
DCLAK11 was a multi-targeted kinase inhibitor that primarily inhibited the EGFR, HER2, and VEGFR2 tyrosine kinases with IC50 value of 6.5, 18, and 31 nmol/L, respectively. DCLAK11 potently inhibited the proliferation of EGFR- and HER2-driven cancer cells: its IC50 value was 12 and 22 nmol/L, respectively, in HCC827 and HCC4006 cells with EGFR exon deletions, and 19 and 81 nmol/L, respectively, in NCI-N87 and BT474 cells with HER2 amplification. Consistently, DCLAK11 blocked the EGFR and HER2 signaling in cancer cells with either an EGFR or a HER2 aberration. Furthermore, DCLAK11 effectively induced EGFR/HER2–driven cell apoptosis. Moreover, DCLAK11 exhibited anti-angiogenic activity, as shown by its inhibitory effect on the proliferation, migration and tube formation of human umbilical vascular endothelial cells and the microvessel outgrowth of rat aortic rings.
Conclusions:
DCLAK11 is a multi-targeted kinase inhibitor with remarkable potency against tyrosine kinases EGFR, HER2 and VEGFR2, which confirms its potent anti-cancer activity in EGFR- and HER2-addicted cancers and its anti-angiogenic activity.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Hunter T . Tyrosine phosphorylation: thirty years and counting. Curr Opin Cell Biol 2009; 21: 140–6.
Hanahan D, Weinberg RA . The hallmarks of cancer. Cell 2000; 100: 57–70.
Luo J, Solimini NL, Elledge SJ . Principles of cancer therapy: oncogene and non-oncogene addiction. Cell 2009; 136: 823–37.
Herbst RS, Heymach JV, Lippman SM . Lung cancer. N Engl J Med 2008; 359: 1367–80.
Seshacharyulu P, Ponnusamy MP, Haridas D, Jain M, Ganti AK, Batra SK . Targeting the EGFR signaling pathway in cancer therapy. Expert Opin Ther Targets 2012; 16: 15–31.
Bose R1, Kavuri SM, Searleman AC, Shen W, Shen D, Koboldt DC, et al. Activating HER2 mutations in HER2 gene amplification negative breast cancer. Cancer Discov 2013; 3: 224–37.
Lee J, Ou SH . Towards the goal of personalized medicine in gastric cancer--time to move beyond HER2 inhibition. Part I: Targeting receptor tyrosine kinase gene amplification. Discov Med 2013; 15: 333–41.
Stephens P1, Hunter C, Bignell G, Edkins S, Davies H, Teague J, et al. Lung cancer: intragenic ERBB2 kinase mutations in tumours. Nature 2004; 431: 525–6.
Shawver LK, Slamon D, Ullrich A . Smart drugs: tyrosine kinase inhibitors in cancer therapy. Cancer Cell 2002; 1: 117–23.
Hubbard SR . Protein tyrosine kinases: autoregulation and smallmolecule inhibition. Curr Opin Struct Biol 2002; 12: 735–41.
Huang M, Shen A, Ding J, Geng M . Molecularly targeted cancer therapy: some lessons from the past decade. Trends Pharmacol Sci 2014; 35: 41–50.
Gschwind A, Fischer OM, Ullrich A . The discovery of receptor tyrosine kinases: targets for cancer therapy. Nat Rev Cancer 2004; 4: 361–70.
Baselga J, Albanell J . Epithelial growth factor receptor interacting agents. Hematol Oncol Clin North Am 2002; 16: 1041–63.
Grandis JR, Sok JC . Signaling through the epidermal growth factor receptor during the development of malignancy. Pharmacol Ther 2004; 102: 37–46.
Tzahar E1, Waterman H, Chen X, Levkowitz G, Karunagaran D, Lavi S, et al. A hierarchical network of interreceptor interactions determines signal transduction by Neu differentiation factor/neuregulin and epidermal growth factor. Mol Cell Biol 1996; 16: 5276–87
Graus-Porta D, Beerli RR, Daly JM, Hynes NE . ErbB-2, the preferred heterodimerization partner of all ErbB receptors, is a mediator of lateral signaling. EMBO J 1997; 16: 1647–55.
Levkowitz G, Alroy I, Klapper L, et al. Diversification of Neu differentiation factor and epidermal growth factor signaling by combinatorial receptor interactions. EMBO J 1996; 15: 2452–67.
Knuefermann C, Lu Y, Liu B, Jin W, Liang K, Wu L, et al. HER2/PI-3K/Akt activation leads to a multidrug resistance in human breast adenocarcinoma cells. Oncogene 2003; 22: 3205–12.
Folkman J . Tumor angiogenesis: therapeutic implications. N Engl J Med 1971; 285: 1182–6.
Folkman J . Endothelial cells and angiogenic growth factors in cancer growth and metastasis. Introduction. Cancer Metastasis Rev 1990; 9: 171–4.
Rini BI . Vascular endothelial growth factor-targeted therapy in renal cell carcinoma: current status and future directions. Clin Cancer Res 2007; 13: 1098–106.
Ferrara N, Davis-Smyth T . The biology of vascular endothelial growth factor. Endocr Rev 1997; 18: 4–25.
Bergers G, Benjamin LE . Tumorigenesis and the angiogenic switch. Nat Rev Cancer 2003; 3: 401–10
Rini BI . Vascular endothelial growth factor-targeted therapy in renal cell carcinoma: current status and future directions. Clin Cancer Res 2007; 13: 1098–106.
Bhargava P, Robinson MO . Development of second generation VEGFR tyrosine kinase inhibitors: current status. Curr Oncol Rep 2011; 13: 103–11.
Viloria-Petit A, Crombet T, Jothy S, Hicklin D, Bohlen P, Schlaeppi JM, et al. Acquired resistance to the antitumor effect of the epidermal growth factor receptor-blocking antibodies in vivo: a role for altered tumor angiogenesis. Cancer Res 2001; 61: 5090–101.
Ferrara N, Kerbel RS . Angiogenesis as a therapeutic target. Nature 2005; 438: 967–74.
Ciardiello F, Bianco R, Damiano V, Fontanini G, Caputo R, Pomatico G, et al. Antiangiogenic and antitumor activity of anti-epidermal growth factor receptor C225 monoclonal antibody in combination with vascular endothelial growth factor antisense oligonucleotide in human GEO colon cancer cells. Clin Cancer Res 2000; 6: 3739–47.
Jung YD, Mansfield PF, Akagi M, Takeda A, Liu W, Bucana CD, et al. Effects of combination anti-vascular endothelial growth factor receptor and anti-epidermal growth factor receptor therapies on the growth of gastric cancer in a nude mouse model. Eur J Cancer 2002; 38: 1133–40
Yazici S, Kim SJ, Busby JE, He J, Thaker P, Yokoi K, et al. Dual inhibition of the epidermal growth factor and vascular endothelial growth factor phosphorylation for antivascular therapy of human prostate cancer in the prostate of nude mice. Prostate 2005; 65: 203–15.
Herbst RS, Johnson DH, Mininberg E, Carbone DP, Henderson T, Kim ES, et al. Phase I/II trial evaluating the antivascular endothelial growth factor monoclonal antibody bevacizumab in combination with the HER-1/epidermal growth factor receptor tyrosine kinase inhibitor erlotinib for patients with recurrent non-small-cell lung cancer. J Clin Oncol 2005; 23: 2544–55.
Guo XN, Zhong L, Zhang XH, Zhao WM, Zhang XW, Lin LP, et al. Evaluation of active recombinant catalytic domain of human ErbB-2 tyrosine kinase, and suppression of activity by a naturally derived inhibitor, ZH-4B. Biochim Biophys Acta 2004; 1673: 186–93.
Yarden Y, Pines G . The ERBB network: at last, cancer therapy meets systems biology. Nat Rev Cancer 2012; 12: 553–63.
Ciardiello F, Caputo R, Damiano V, Caputo R, Troiani T, Vitagliano D, et al. Antitumor effects of ZD6474, a small molecule vascular endothelial growth factor receptor tyrosine kinase inhibitor, with additional activity against epidermal growth factor receptor tyrosine kinase. Clin Cancer Res 2003; 9: 1546–56.
Tortora G, Ciardiello F, Gasparini G . Combined targeting of EGFR dependent and VEGF-dependent pathways: rationale, preclinical studies and clinical applications. Nat Clin Pract Oncol 2008; 5: 521–30.
Vailhé B, Vittet D, Feige JJ . In vitro models of vasculogenesis and angiogenesis. Lab Investig 2001; 81: 439–52.
Acknowledgements
We gratefully acknowledge financial support from the National Natural Science Foundation of China for Innovation Research Group (No 81321092), the National Marine “863” project (No 2013AA092902), and the National Natural Science Foundation of China Grants (91229204).
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Scheme 1 is available at the Acta Pharmacologica Sinica's website.
Supplementary information
Materials and methods (download DOCX )
Synthesis of the compound DCLAK11 (DOCX 86 kb)
Rights and permissions
About this article
Cite this article
Guo, Xb., Chen, Xj., Tong, Lj. et al. DCLAK11, a multi-tyrosine kinase inhibitor, exhibits potent antitumor and antiangiogenic activity in vitro. Acta Pharmacol Sin 36, 1266–1276 (2015). https://doi.org/10.1038/aps.2015.25
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/aps.2015.25
