Investigational New Drugs

, Volume 29, Issue 6, pp 1132–1142 | Cite as

Combined anticancer effects of sphingosine kinase inhibitors and sorafenib

  • Vladimir Beljanski
  • Christian Knaak
  • Yan Zhuang
  • Charles D. SmithEmail author


The pro-apoptotic lipid sphingosine is phosphorylated by sphingosine kinases 1 and 2 (SK1 and SK2) to generate the mitogenic lipid sphingosine-1-phosphate (S1P). We previously reported that inhibition of SK activity delays tumor growth in a mouse mammary adenocarcinoma model. Because SK inhibitors and the multikinase inhibitor sorafenib both suppress the MAP kinase pathway, we hypothesized that their combination may provide enhanced inhibition of tumor growth. Therefore, we evaluated the effects of two SK inhibitors, ABC294640 (a SK2-specific inhibitor) and ABC294735 (a dual SK1/SK2 inhibitor), alone and in combination with sorafenib on human pancreatic adenocarcinoma (Bxpc-3) and kidney carcinoma (A-498) cells in vitro and in vivo. Exposure of either Bxpc-3 or A-498 cells to combinations of ABC294640 and sorafenib or ABC294735 and sorafenib resulted in synergistic cytotoxicity, associated with activation of caspases 3/7 and DNA fragmentation. Additionally, strong decreases in ERK phosphorylation were observed in Bxpc-3 and A-498 cells exposed to either the sorafenib/ABC294640 or the sorafenib/ABC294735 combination. Oral administration of either ABC294640 or ABC294735 to mice led to a delay in tumor growth in both xenograft models without overt toxicity to the animals. Tumor growth delay was potentiated by co-administration of sorafenib. These studies show that combination of an SK inhibitor with sorafenib causes synergistic inhibition of cell growth in vitro, and potentiates antitumor activity in vivo. Thus, a foundation is established for clinical trials evaluating the efficacy of combining these signaling inhibitors.


Targeted therapy Sphingosine kinase Sorafenib Apoptosis MAPK pathway 



Funding was provided by National Institutes of Health grant R01 CA122226.


  1. 1.
    Kessler T, Bayer M, Schwoppe C, Liersch R, Mesters RM, Berdel WE (2010) Compounds in clinical Phase III and beyond. Recent Results Cancer Res 180:137–163PubMedCrossRefGoogle Scholar
  2. 2.
    Wymann MP, Schneiter R (2008) Lipid signalling in disease. Nat Rev Mol Cell Biol 9:162–176PubMedCrossRefGoogle Scholar
  3. 3.
    Tani M, Ito M, Igarashi Y (2007) Ceramide/sphingosine/sphingosine 1-phosphate metabolism on the cell surface and in the extracellular space. Cell Signal 19:229–237PubMedCrossRefGoogle Scholar
  4. 4.
    Alemany R, van Koppen CJ, Danneberg K, Ter Braak M, Meyer Zu Heringdorf D (2007) Regulation and functional roles of sphingosine kinases. Naunyn-Schmiedeberg’s Arch Pharmacol 374:413–428CrossRefGoogle Scholar
  5. 5.
    Baran Y, Salas A, Senkal CE, Gunduz U, Bielawski J, Obeid LM, Ogretmen B (2007) Alterations of ceramide/sphingosine 1-phosphate rheostat involved in the regulation of resistance to imatinib-induced apoptosis in K562 human chronic myeloid leukemia cells. J Biol Chem 282:10922–10934PubMedCrossRefGoogle Scholar
  6. 6.
    Le Scolan E, Pchejetski D, Banno Y, Denis N, Mayeux P, Vainchenker W, Levade T, Moreau-Gachelin F (2005) Overexpression of sphingosine kinase 1 is an oncogenic event in erythroleukemic progression. Blood 106:1808–1816PubMedCrossRefGoogle Scholar
  7. 7.
    Safadi-Chamberlain F, Wang LP, Payne SG, Lim CU, Stratford S, Chavez JA, Fox MH, Spiegel S, Summers SA (2005) Effect of a membrane-targeted sphingosine kinase 1 on cell proliferation and survival. Biochem J 388:827–834PubMedCrossRefGoogle Scholar
  8. 8.
    Sarkar S, Maceyka M, Hait NC, Paugh SW, Sankala H, Milstien S, Spiegel S (2005) Sphingosine kinase 1 is required for migration, proliferation and survival of MCF-7 human breast cancer cells. FEBS Lett 579:5313–5317PubMedCrossRefGoogle Scholar
  9. 9.
    French KJ, Schrecengost RS, Lee BD, Zhuang Y, Smith SN, Eberly JL, Yun JK, Smith CD (2003) Discovery and evaluation of inhibitors of human sphingosine kinase. Cancer Res 63:5962–5969PubMedGoogle Scholar
  10. 10.
    French KJ, Upson JJ, Keller SN, Zhuang Y, Yun JK, Smith CD (2006) Antitumor activity of sphingosine kinase inhibitors. J Pharmacol Exp Ther 318:596–603PubMedCrossRefGoogle Scholar
  11. 11.
    French KJ, Zhuang Y, Maines LW, Gao P, Wang W, Beljanski V, Upson JJ, Green CL, Keller SN, Smith CD (2010) Pharmacology and antitumor activity of ABC294640, a selective inhibitor of sphingosine kinase-2. J Pharmacol Exp Ther 333:129–139PubMedCrossRefGoogle Scholar
  12. 12.
    Maines LW, French KJ, Wolpert EB, Antonetti DA, Smith CD (2006) Pharmacologic manipulation of sphingosine kinase in retinal endothelial cells: implications for angiogenic ocular diseases. Invest Ophthalmol Vis Sci 47:5022–5031PubMedCrossRefGoogle Scholar
  13. 13.
    Young A, Lyons J, Miller AL, Phan VT, Alarcon IR, McCormick F (2009) Ras signaling and therapies. Adv Cancer Res 102:1–17PubMedCrossRefGoogle Scholar
  14. 14.
    Wilhelm S, Carter C, Lynch M, Lowinger T, Dumas J, Smith RA, Schwartz B, Simantov R, Kelley S (2006) Discovery and development of sorafenib: a multikinase inhibitor for treating cancer. Nat Rev Drug Discov 5:835–844PubMedCrossRefGoogle Scholar
  15. 15.
    Wilhelm SM, Adnane L, Newell P, Villanueva A, Llovet JM, Lynch M (2008) Preclinical overview of sorafenib, a multikinase inhibitor that targets both Raf and VEGF and PDGF receptor tyrosine kinase signaling. Mol Cancer Ther 7:3129–3140PubMedCrossRefGoogle Scholar
  16. 16.
    Sharma A, Trivedi NR, Zimmerman MA, Tuveson DA, Smith CD, Robertson GP (2005) Mutant V599EB-Raf regulates growth and vascular development of malignant melanoma tumors. Cancer Res 65:2412–2421PubMedCrossRefGoogle Scholar
  17. 17.
    Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82:1107–1112PubMedCrossRefGoogle Scholar
  18. 18.
    Chou TC, Talalay P (1984) Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27–55PubMedCrossRefGoogle Scholar
  19. 19.
    Beljanski V, Knaak C, Smith CD. A novel sphingosine kinase inhibitor induces autophagy in tumor cells. J Pharmacol Exp Ther 333:454-464Google Scholar
  20. 20.
    Juang SH, Lung CC, Hsu PC, Hsu KS, Li YC, Hong PC, Shiah HS, Kuo CC, Huang CW, Wang YC, Huang L, Chen TS, Chen SF, Fu KC, Hsu CL, Lin MJ, Chang CJ, Ashendel CL, Chan TC, Chou KM, Chang JY (2007) D-501036, a novel selenophene-based triheterocycle derivative, exhibits potent in vitro and in vivo antitumoral activity which involves DNA damage and ataxia telangiectasia-mutated nuclear protein kinase activation. Mol Cancer Ther 6:193–202PubMedCrossRefGoogle Scholar
  21. 21.
    Van Quaquebeke E, Mahieu T, Dumont P, Dewelle J, Ribaucour F, Simon G, Sauvage S, Gaussin JF, Tuti J, El Yazidi M, Van Vynckt F, Mijatovic T, Lefranc F, Darro F, Kiss R (2007) 2, 2, 2-Trichloro-N-({2-[2-(dimethylamino)ethyl]-1, 3-dioxo-2, 3-dihydro-1H-be nzo[de]isoquinolin- 5-yl}carbamoyl)acetamide (UNBS3157), a novel nonhematotoxic naphthalimide derivative with potent antitumor activity. J Med Chem 50:4122–4134PubMedCrossRefGoogle Scholar
  22. 22.
    Sridhar SS, Hedley D, Siu LL (2005) Raf kinase as a target for anticancer therapeutics. Mol Cancer Ther 4:677–685PubMedCrossRefGoogle Scholar
  23. 23.
    Rodriguez-Viciana P, Oses-Prieto J, Burlingame A, Fried M, McCormick F (2006) A phosphatase holoenzyme comprised of Shoc2/Sur8 and the catalytic subunit of PP1 functions as an M-Ras effector to modulate Raf activity. Mol Cell 22:217–230PubMedCrossRefGoogle Scholar
  24. 24.
    Klionsky DJ, Abeliovich H, Agostinis P, Agrawal DK, Aliev G, Askew DS, Baba M, Baehrecke EH, Bahr BA, Ballabio A, Bamber BA, Bassham DC, Bergamini E, Bi X, Biard-Piechaczyk M, Blum JS, Bredesen DE, Brodsky JL, Brumell JH, Brunk UT, Bursch W, Camougrand N, Cebollero E, Cecconi F, Chen Y, Chin LS, Choi A, Chu CT, Chung J, Clarke PG, Clark RS, Clarke SG, Clave C, Cleveland JL, Codogno P, Colombo MI, Coto-Montes A, Cregg JM, Cuervo AM, Debnath J, Demarchi F, Dennis PB, Dennis PA, Deretic V, Devenish RJ, Di Sano F, Dice JF, Difiglia M, Dinesh-Kumar S, Distelhorst CW, Djavaheri-Mergny M, Dorsey FC, Droge W, Dron M, Dunn WA Jr, Duszenko M, Eissa NT, Elazar Z, Esclatine A, Eskelinen EL, Fesus L, Finley KD, Fuentes JM, Fueyo J, Fujisaki K, Galliot B, Gao FB, Gewirtz DA, Gibson SB, Gohla A, Goldberg AL, Gonzalez R, Gonzalez-Estevez C, Gorski S, Gottlieb RA, Haussinger D, He YW, Heidenreich K, Hill JA, Hoyer-Hansen M, Hu X, Huang WP, Iwasaki A, Jaattela M, Jackson WT, Jiang X, Jin S, Johansen T, Jung JU, Kadowaki M, Kang C, Kelekar A, Kessel DH, Kiel JA, Kim HP, Kimchi A, Kinsella TJ, Kiselyov K, Kitamoto K, Knecht E et al (2008) Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 4:151–175PubMedGoogle Scholar
  25. 25.
    Facchini G, Perri F, Caraglia M, Pisano C, Striano S, Marra L, Fiore F, Aprea P, Pignata S, Iaffaioli RV (2009) New treatment approaches in renal cell carcinoma. Anticancer Drugs 20:893–900PubMedCrossRefGoogle Scholar
  26. 26.
    Motzer RJ, Bander NH, Nanus DM (1996) Renal-cell carcinoma. N Engl J Med 335:865–875PubMedCrossRefGoogle Scholar
  27. 27.
    Gkialas IK, Papadopoulos G (2009) New therapeutic approaches in the management of metastatic renal cell carcinoma. J BUON 14:399–404PubMedGoogle Scholar
  28. 28.
    Lee T, Kim J, Sohn U (2002) Sphingosylphosphorylcholine-induced contraction of feline ileal smooth muscle cells is mediated by Galphai3 protein and MAPK. Cell Signal 14:989–997PubMedCrossRefGoogle Scholar
  29. 29.
    Saddoughi SA, Song P, Ogretmen B (2008) Roles of bioactive sphingolipids in cancer biology and therapeutics. Subcell Biochem 49:413–440PubMedCrossRefGoogle Scholar
  30. 30.
    Kolesnick R, Fuks Z (2003) Radiation and ceramide-induced apoptosis. Oncogene 22:5897–5906PubMedCrossRefGoogle Scholar
  31. 31.
    Spiegel S, Kolesnick R (2002) Sphingosine 1-phosphate as a therapeutic agent. Leukemia 16:1596–1602PubMedCrossRefGoogle Scholar
  32. 32.
    Bolz SS, Vogel L, Sollinger D, Derwand R, Boer C, Pitson SM, Spiegel S, Pohl U (2003) Sphingosine kinase modulates microvascular tone and myogenic responses through activation of RhoA/Rho kinase. Circulation 108:342–347PubMedCrossRefGoogle Scholar
  33. 33.
    Stadler WM (2005) Targeted agents for the treatment of advanced renal cell carcinoma. Cancer 104:2323–2333PubMedCrossRefGoogle Scholar
  34. 34.
    Chadha KS, Khoury T, Yu J, Black JD, Gibbs JF, Kuvshinoff BW, Tan D, Brattain MG, Javle MM (2006) Activated Akt and Erk expression and survival after surgery in pancreatic carcinoma. Ann Surg Oncol 13:933–939PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Vladimir Beljanski
    • 1
  • Christian Knaak
    • 1
  • Yan Zhuang
    • 2
  • Charles D. Smith
    • 1
    • 2
    Email author
  1. 1.Drug Discovery Core, Hollings Cancer Center and Department of Pharmaceutical and Biomedical SciencesMedical University of South CarolinaCharlestonUSA
  2. 2.Apogee Biotechnology CorporationHummelstownUSA

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