Investigational New Drugs

, Volume 26, Issue 3, pp 223–232 | Cite as

A novel quinoline, MT477: suppresses cell signaling through Ras molecular pathway, inhibits PKC activity, and demonstrates in vivo anti-tumor activity against human carcinoma cell lines

  • Piotr Jasinski
  • Brandon Welsh
  • Jorge Galvez
  • David Land
  • Pawel Zwolak
  • Lori Ghandi
  • Kaoru Terai
  • Arkadiusz Z. Dudek


MT477 is a novel thiopyrano[2,3-c]quinoline that has been identified using molecular topology screening as a potential anticancer drug with a high activity against protein kinase C (PKC) isoforms. The objective of the present study was to determine the mechanism of action of MT477 and its activity against human cancer cell lines. MT477 interfered with PKC activity as well as phosphorylation of Ras and ERK1/2 in H226 human lung carcinoma cells. It also induced poly-caspase-dependent apoptosis. MT477 had a dose-dependent (0.006 to 0.2 mM) inhibitory effect on cellular proliferation of H226, MCF-7, U87, LNCaP, A431 and A549 cancer cell lines as determined by in vitro proliferation assays. Two murine xenograft models of human A431 and H226 lung carcinoma were used to evaluate tumor response to intraperitoneal administration of MT477 (33 μg/kg, 100 μg/kg, and 1 mg/kg). Tumor growth was inhibited by 24.5% in A431 and 43.67% in H226 xenografts following MT477 treatment, compared to vehicle controls (p < 0.05). In conclusion, our empirical findings are consistent with molecular modeling of MT477’s activity against PKC. We also found, however, that its mechanism of action occurs through suppressing Ras signaling, indicating that its effects on apoptosis and tumor growth in vivo may be mediated by Ras as well as PKC. We propose, therefore, that MT477 warrants further development as an anticancer drug.


MT477 Protein Kinase C Ras-MEK-ERK pathway inhibition Caspase-dependent apoptosis New drug development 



Protein kinase C


Glycogen synthase kinase-3β


3-(4,5-dimethylthiazol-2y1)-2,5-diphenyltetrazolium bromide


Fluromethyl ketone




Valylalanylaspartic acid


Fluoromethyl ketone



This study was partially supported by Experimental Therapeutics Fund from University of Minnesota (we are grateful to Audrey and Denis Anderson for ongoing support for this Fund) and a grant from Medisyn Technologies. We would like to thank Michael Franklin for editorial support.


  1. 1.
    Galvez J, Garcia-Domenech R, de Julian-Ortiz JV, Soler R (1995) Topological approach to drug design. J Chem Inf Comput Sci 35(2):272–284PubMedCrossRefGoogle Scholar
  2. 2.
    Llompart J, Galvez J, Pal K (2006) Inventors; US Patent 20060014770Google Scholar
  3. 3.
    Goekjian PG, Jirousek MR (2001) Protein kinase C inhibitors as novel anticancer drugs. Expert Opin Investig Drugs 10(12):2117–2140PubMedCrossRefGoogle Scholar
  4. 4.
    Yoshiji H, Kuriyama S, Ways DK et al (1999) Protein kinase C lies on the signaling pathway for vascular endothelial growth factor-mediated tumor development and angiogenesis. Cancer Res 59(17):4413–4418PubMedGoogle Scholar
  5. 5.
    Hans CP, Weisenburger DD, Greiner TC et al (2005) Expression of PKC-beta or cyclin D2 predicts for inferior survival in diffuse large B-cell lymphoma. Mod Path 18(10):1377–1384CrossRefGoogle Scholar
  6. 6.
    da Rocha AB, Mans DR, Regner A, Schwartsmann G (2002) Targeting protein kinase C: new therapeutic opportunities against high-grade malignant gliomas? Oncologist 7(1):17–33PubMedCrossRefGoogle Scholar
  7. 7.
    Gokmen-Polar Y, Murray NR, Velasco MA, Gatalica Z, Fields AP (2001) Elevated protein kinase C betaII is an early promotive event in colon carcinogenesis. Cancer Res 61(4):1375–1381PubMedGoogle Scholar
  8. 8.
    Lahn M, Kohler G, Sundell K et al (2004) Protein kinase C alpha expression in breast and ovarian cancer. Oncology 67(1):1–10PubMedCrossRefGoogle Scholar
  9. 9.
    Regala RP, Weems C, Jamieson L et al (2005) Atypical protein kinase C iota is an oncogene in human non-small cell lung cancer. Cancer Res 65(19):8905–8911PubMedCrossRefGoogle Scholar
  10. 10.
    Mandil R, Ashkenazi E, Blass M et al (2001) Protein kinase Calpha and protein kinase Cdelta play opposite roles in the proliferation and apoptosis of glioma cells. Cancer Res 61(11):4612–4619PubMedGoogle Scholar
  11. 11.
    Liu JF, Crepin M, Liu JM, Barritault D, Ledoux D (2002) FGF-2 and TPA induce matrix metalloproteinase-9 secretion in MCF-7 cells through PKC activation of the Ras/ERK pathway. Biochem Biophys Res Commun 293(4):1174–1182PubMedCrossRefGoogle Scholar
  12. 12.
    Villalonga P, Lopez-Alcala C, Chiloeches A et al (2002) Calmodulin prevents activation of Ras by PKC in 3T3 fibroblasts. J Biol Chem 277(40):37929–37935PubMedCrossRefGoogle Scholar
  13. 13.
    Zhang J, Anastasiadis PZ, Liu Y, Thompson EA, Fields AP (2004) Protein kinase C (PKC) betaII induces cell invasion through a Ras/Mek-, PKC iota/Rac 1-dependent signaling pathway. J Biol Chem 279(21):22118–22123PubMedCrossRefGoogle Scholar
  14. 14.
    Dudek AZ, Zwolak P, Jasinski P et al (2007) Protein kinase C-beta inhibitor enzastaurin (LY317615.HCI) enhances radiation control of murine breast cancer in an orthotopic model of bone metastasis. Invest New Drugs, Sep 6.Google Scholar
  15. 15.
    Slupsky JR, Kamiguti AS, Harris RJ, Cawley JC, Zuzel M (2007) Central role of protein kinase Cepsilon in constitutive activation of ERK1/2 and Rac1 in the malignant cells of hairy cell leukemia. Am J Pathol 170(2):745–754PubMedCrossRefGoogle Scholar
  16. 16.
    Song MS, Park YK, Lee JH, Park K (2001) Induction of glucose-regulated protein 78 by chronic hypoxia in human gastric tumor cells through a protein kinase C-epsilon/ERK/AP-1 signaling cascade. Cancer Res 61(22):8322–8330PubMedGoogle Scholar
  17. 17.
    Kolch W, Heidecker G, Kochs G et al (1993) Protein kinase C alpha activates RAF-1 by direct phosphorylation. Nature 364(6434):249–252PubMedCrossRefGoogle Scholar
  18. 18.
    Goode N, Hughes K, Woodgett JR, Parker PJ (1992) Differential regulation of glycogen synthase kinase-3 beta by protein kinase C isotypes. J Biol Chem 267(24):16878–16882PubMedGoogle Scholar
  19. 19.
    Downward J (2003) Targeting RAS signalling pathways in cancer therapy. Nat Rev 3(1):11–22CrossRefGoogle Scholar
  20. 20.
    Johnson L, Mercer K, Greenbaum D et al (2001) Somatic activation of the K-ras oncogene causes early onset lung cancer in mice. Nature 410(6832):1111–1116PubMedCrossRefGoogle Scholar
  21. 21.
    Shields JM, Pruitt K, McFall A, Shaub A, Der CJ (2000) Understanding Ras: ‘it ain’t over ‘til it’s over’. Trends Cell Biol 10(4):147–154PubMedCrossRefGoogle Scholar
  22. 22.
    Downward J (2006) Signal transduction. Prelude to an anniversary for the RAS oncogene. Science 314(5798):433–434PubMedCrossRefGoogle Scholar
  23. 23.
    D’Argenio D (1997) Adapt II User’s Guide: Pharmacokinetic/Pharmacodynamic System Analysis Software. Los Angeles: Biomedical Simulations ResourceGoogle Scholar
  24. 24.
    Okudela K, Hayashi H, Ito T et al (2004) K-ras gene mutation enhances motility of immortalized airway cells and lung adenocarcinoma cells via Akt activation: possible contribution to non-invasive expansion of lung adenocarcinoma. Am J Pathol 164(1):91–100PubMedGoogle Scholar
  25. 25.
    Toulany M, Dittmann K, Kruger M, Baumann M, Rodemann HP (2005) Radioresistance of K-Ras mutated human tumor cells is mediated through EGFR-dependent activation of PI3K-AKT pathway. Radiother Oncol 76(2):143–150PubMedCrossRefGoogle Scholar
  26. 26.
    Gorczyca W, Bigman K, Mittelman A et al (1993) Induction of DNA strand breaks associated with apoptosis during treatment of leukemias. Leukemia 7(5):659–670PubMedGoogle Scholar
  27. 27.
    Gorczyca W, Gong J, Ardelt B, Traganos F, Darzynkiewicz Z (1993) The cell cycle related differences in susceptibility of HL-60 cells to apoptosis induced by various antitumor agents. Cancer Res 53(13):3186–3192PubMedGoogle Scholar
  28. 28.
    Wyllie AH, Kerr JF, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68:251–306PubMedCrossRefGoogle Scholar
  29. 29.
    Scholz C, Wieder T, Starck L et al (2005) Arsenic trioxide triggers a regulated form of caspase-independent necrotic cell death via the mitochondrial death pathway. Oncogene 24(11):1904–1913PubMedCrossRefGoogle Scholar
  30. 30.
    Qiu RG, Chen J, Kirn D, McCormick F, Symons M (1995) An essential role for Rac in Ras transformation. Nature 374(6521):457–459PubMedCrossRefGoogle Scholar
  31. 31.
    Clark JA, Black AR, Leontieva OV et al (2004) Involvement of the ERK signaling cascade in protein kinase C-mediated cell cycle arrest in intestinal epithelial cells. J Biol Chem 279(10):9233–9247PubMedCrossRefGoogle Scholar
  32. 32.
    Basu A, Tu H (2005) Activation of ERK during DNA damage-induced apoptosis involves protein kinase Cdelta. Biochem Biophys Res Commun 334(4):1068–1073PubMedCrossRefGoogle Scholar
  33. 33.
    Muscella A, Greco S, Elia MG, Storelli C, Marsigliante S (2004) Differential signalling of purinoceptors in HeLa cells through the extracellular signal-regulated kinase and protein kinase C pathways. J Cell Physiol 200(3):428–439PubMedCrossRefGoogle Scholar
  34. 34.
    Weisstein JS, Majeska RJ, Klein MJ, Einhorn TA (2001) Detection of c-fos expression in benign and malignant musculoskeletal lesions. J Orthop Res 19(3):339–345PubMedCrossRefGoogle Scholar
  35. 35.
    Wang ZQ, Grigoriadis AE, Mohle-Steinlein U, Wagner EF (1991) A novel target cell for c-fos-induced oncogenesis: development of chondrogenic tumours in embryonic stem cell chimeras. EMBO J 10(9):2437–2450PubMedGoogle Scholar
  36. 36.
    Zajchowski DA, Bartholdi MF, Gong Y et al (2001) Identification of gene expression profiles that predict the aggressive behavior of breast cancer cells. Cancer Res 61(13):5168–5178PubMedGoogle Scholar
  37. 37.
    Kustikova O, Kramerov D, Grigorian M et al (1998) Fra-1 induces morphological transformation and increases in vitro invasiveness and motility of epithelioid adenocarcinoma cells. Mol Cell Biol 18(12):7095–7105PubMedGoogle Scholar
  38. 38.
    Lewis TS, Shapiro PS, Ahn NG (1998) Signal transduction through MAP kinase cascades. Adv Cancer Res 74:49–139PubMedGoogle Scholar
  39. 39.
    Chen Z, Gibson TB, Robinson F et al (2001) MAP kinases. Chem Rev 101(8):2449–2476PubMedCrossRefGoogle Scholar
  40. 40.
    Celano P, Berchtold CM, Mabry M et al (1993) Induction of markers of normal differentiation in human colon carcinoma cells by the v-rasH oncogene. Cell Growth Differ 4(4):341–347PubMedGoogle Scholar
  41. 41.
    Wang X, Studzinski GP (2001) Activation of extracellular signal-regulated kinases (ERKs) defines the first phase of 1,25-dihydroxyvitamin D3-induced differentiation of HL60 cells. J Cell Biochem 80(4):471–482PubMedCrossRefGoogle Scholar
  42. 42.
    Cheng M, Zhen E, Robinson MJ, Ebert D, Goldsmith E, Cobb MH (1996) Characterization of a protein kinase that phosphorylates serine 189 of the mitogen-activated protein kinase homolog ERK3. J Biol Chem 271(20):12057–12062PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Piotr Jasinski
    • 1
    • 2
  • Brandon Welsh
    • 1
  • Jorge Galvez
    • 3
  • David Land
    • 4
  • Pawel Zwolak
    • 1
  • Lori Ghandi
    • 1
  • Kaoru Terai
    • 1
  • Arkadiusz Z. Dudek
    • 1
  1. 1.Division of Hematology, Oncology and Transplantation, Department of MedicineUniversity of MinnesotaMinneapolisUSA
  2. 2.Department of PathophysiologyMedical University of ViennaViennaAustria
  3. 3.Molecular Connectivity & Drug Design Research Unit, Department of Physical ChemistryUniversity of ValenciaValenciaSpain
  4. 4.Medisyn Technologies, Inc.MinnetonkaUSA

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