Journal of Neuro-Oncology

, Volume 110, Issue 2, pp 163–177 | Cite as

Mechanism of anti-glioma activity and in vivo efficacy of the cannabinoid ligand KM-233

  • Steven N. Gurley
  • Ammaar H. Abidi
  • Patrick Allison
  • Peihong Guan
  • Christopher Duntsch
  • Jon H. Robertson
  • Stanley D. Kosanke
  • Stephen T. Keir
  • Darell D. Bigner
  • Andrea J. Elberger
  • Bob M. MooreII
Laboratory Investigation


Glioblastoma multiforme (GBM) is the most common and devastating form of primary central nervous system malignancy. The prognosis for patients diagnosed with GBM is poor, having a median survival rate of 12–15 months. Despite modern advances in the development of antineoplastic agents, the efficacy of newer anti-cancer agents in the treatment of GBM is yet to be determined. Thus, there remains a significant unmet need for new therapeutic strategies against GBM. A promising chemotherapeutic intervention has emerged from studies of cannabinoid receptor agonists wherein tetrahydrocannabinol has been the most extensively studied. The novel cannabinoid ligand KM-233 was developed as a lead platform for future optimization of biopharmaceutical properties of classical based cannabinoid ligands. Treatment of U87MG human GBM cells with KM-233 caused a time dependent change in the phosphorylation profiles of MEK, ERK1/2, Akt, BAD, STAT3, and p70S6K. Almost complete mitochondrial depolarization was observed 6 h post-treatment followed by a rapid increase in cleaved caspase 3 and significant cytoskeletal contractions. Treatment with KM-233 also resulted in a redistribution of the Golgi-endoplasmic reticulum structures. Dose escalation studies in the orthotopic model using U87MG cells revealed an 80 % reduction in tumor size after 12 mg/kg daily dosing for 20 days. The evaluation of KM-233 against primary tumor tissue in the side flank model revealed a significant decrease in the rate of tumor growth. These findings indicate that structural refinement of KM-233 to improve its biopharmaceutical properties may lead to a novel and efficacious treatment for GBM.


Brain malignancy Cannabinoid Chemotherapeutic Glioma Therapy 


  1. 1.
    Jemal A, Siegel R, Ward E, Hao Y, Xu Z, Thun TJ (2009) Cancer statistics, 2009. Cancer J Clin 59:225–249CrossRefGoogle Scholar
  2. 2.
    Surawicz TS, Davis F, Freels S, Laws ER Jr, Menck HR (1998) Brain tumor survival: results from the National Cancer Data Base. J Neurooncol 40(2):151–160PubMedCrossRefGoogle Scholar
  3. 3.
    Van Meir EG, Hadjipanayis CG, Norden AD, Shu HK, Wen PY, Olson JJ (2010) Exciting new advances in neuro-oncology: the avenue to a cure for Malignant Glioma. Cancer J Clin 60:166–193CrossRefGoogle Scholar
  4. 4.
    Parolaro D, Massi P (2008) Cannabinoids as potential new therapy for the treatment of gliomas. Expert Rev Neurother 8:37–49PubMedCrossRefGoogle Scholar
  5. 5.
    Velasco G, Carracedo A, Blázquez C, Lorente M, Aguado T, Haro A, Sánchez C, Galve-Roperh I, Guzmán M (2007) Cannabinoids and gliomas. Mol Neurobiol 36:60–67PubMedCrossRefGoogle Scholar
  6. 6.
    Sanchez C, Galve-Roperh I, Canova C, Brachet P, Guzman M (1998) Delta9-tetrahydrocannabinol induces apoptosis in C6 glioma cells. FEBS Lett 436(1):6–10PubMedCrossRefGoogle Scholar
  7. 7.
    Duntsch C, Divi MK, Jones T, Zhou Q, Krishnamurthy M, Boehm P, Wood G, Sills A, Moore BM (2006) Safety and efficacy of a novel cannabinoid chemotherapeutic, KM-233, for the treatment of high-grade glioma. J Neurooncol 77:143–152PubMedCrossRefGoogle Scholar
  8. 8.
    Galve-Roperh I, Rueda D, Gómez dPT, Velasco G, Guzmán M (2002) Mechanism of extracellular signal-regulated kinase activation by the CB(1) cannabinoid receptor. Mol Pharmacol 62:1385–1392PubMedCrossRefGoogle Scholar
  9. 9.
    Jacobsson SO, Wallin T, Fowler CJ (2001) Inhibition of rat C6 glioma cell proliferation by endogenous and synthetic cannabinoids. Relative involvement of cannabinoid and vanilloid receptors. J Pharmacol Exp Ther 299(3):951–959PubMedGoogle Scholar
  10. 10.
    Krishnamurthy M, Gurley S, Moore BM (2008) Exploring the substituent effects on a novel series of C1′-dimethyl-aryl Delta8-tetrahydrocannabinol analogs. Bioorg Med Chem 16:6489–6500PubMedCrossRefGoogle Scholar
  11. 11.
    Massi P, Vaccani A, Ceruti S, Colombo A, Abbracchio MP, Parolaro D (2004) Antitumor effects of cannabidiol, a nonpsychoactive cannabinoid, on human glioma cell lines. J Pharmacol Exp Ther 308(3):838–845PubMedCrossRefGoogle Scholar
  12. 12.
    Sanchez C, de Ceballos ML, del Pulgar TG, Rueda D, Corbacho C, Velasco G, Galve-Roperh I, Huffman JW, Ramon y Cajal S, Guzman M (2001) Inhibition of glioma growth in vivo by selective activation of the CB(2) cannabinoid receptor. Cancer Res 61(15):5784–5789PubMedGoogle Scholar
  13. 13.
    Marcu JP, Christian RT, Lau D, Zielinski AJ, Horowitz MP, Lee J, Pakdel A, Allison J, Limbad C, Moore DH, Yount GL, Desprez PY, McAlliste rSD (2010) Cannabidiol enhances the inhibitory effects of delta9-tetrahydrocannabinol on human glioblastoma cell proliferation and survival. Mol Cancer Ther 9:180–189PubMedCrossRefGoogle Scholar
  14. 14.
    Eichele K, Weinzierl U, Ramer R, Brune K, Hinz B (2006) R(+)-methanandamide elicits a cyclooxygenase-2-dependent mitochondrial apoptosis signaling pathway in human neuroglioma cells. Pharm Res 23:90–94PubMedCrossRefGoogle Scholar
  15. 15.
    Blázquez C, González-Feria L, Alvarez L, Haro A, Casanova ML, Guzmán M (2004) Cannabinoids inhibit the vascular endothelial growth factor pathway in gliomas. Cancer Res 64:5617–5623PubMedCrossRefGoogle Scholar
  16. 16.
    Blázquez C, Salazar M, Carracedo A, Lorente M, Egia A, González-Feria L, Haro A, Velasco G, Guzmán M (2008) Cannabinoids inhibit glioma cell invasion by down-regulating matrix metalloproteinase-2 expression. Cancer Res 68:1945–1952PubMedCrossRefGoogle Scholar
  17. 17.
    Carracedo A, Lorente M, Egia A, Blázquez C, García S, Giroux V, Malicet C, Villuendas R, Gironella M, González-Feria L, Piris MA, Iovanna JL, Guzmán M, Velasco G (2006) The stress-regulated protein p8 mediates cannabinoid-induced apoptosis of tumor cells. Cancer Cell 9:301–312Google Scholar
  18. 18.
    Galve-Roperh I, Sanchez C, Cortes ML, del Pulgar TG, Izquierdo M, Guzman M (2000) Anti-tumoral action of cannabinoids: involvement of sustained ceramide accumulation and extracellular signal-regulated kinase activation. Nat Med 6(3):313–319PubMedCrossRefGoogle Scholar
  19. 19.
    Salazar M, Carracedo A, Salanueva IJ, Hernández-Tiedra S, Lorente M, Egia A, Vázquez P, Blázquez C, Torres S, García S, Nowak J, Fimia GM, Piacentin iM, Cecconi F, Pandolf iPP, González-Feria L, Iovanna JL, Guzmán M, Boya P, Velasco G (2009) Cannabinoid action induces autophagy-mediated cell death through stimulation of ER stress in human glioma cells. J Clin Invest 119:1359–1372Google Scholar
  20. 20.
    Lorente M, Torres S, Salazar M, Carracedo A, Hernández-Tiedra S, Rodríguez-Fornés F, García-Taboada E, Meléndez B, Mollejo M, Campos-Martín Y, Lakatosh SA, Barcia J, Guzmán M, Velasco G (2011) Stimulation of the midkine/ALK axis renders glioma cells resistant to cannabinoid antitumoral action. Cell Death Differ 18:959–973PubMedCrossRefGoogle Scholar
  21. 21.
    Galanti G, Fisher T, Kventsel I, Shoham J, Gallily R, Mechoulam R, Lavie G, Amariglio N, Rechavi G, Toren A (2008) Delta 9-tetrahydrocannabinol inhibits cell cycle progression by downregulation of E2F1 in human glioblastoma multiforme cells. Acta Oncol 47:1062–1070PubMedCrossRefGoogle Scholar
  22. 22.
    Guzmán M, Duarte MJ, Blázquez C, Ravina J, Rosa MC, Galve-Roperh I, Sánchez C, Velasco G, González-Feria L (2006) A pilot clinical study of Delta9-tetrahydrocannabinol in patients with recurrent glioblastoma multiforme. Br J Cancer 95:197–203PubMedCrossRefGoogle Scholar
  23. 23.
    Ellert-Miklaszewska A, Grajkowska W, Gabrusiewicz K, Kaminska B, Konarska L (2007) Distinctive pattern of cannabinoid receptor type II (CB2) expression in adult and pediatric brain tumors. Brain Res 1137:161–169PubMedCrossRefGoogle Scholar
  24. 24.
    De Jesús ML, Hostalot C, Garibi JM, Sallés J, Meana JJ, Callado LF (2010) Opposite changes in cannabinoid CB1 and CB2 receptor expression in human gliomas. Neurochem Int 56:829–833PubMedCrossRefGoogle Scholar
  25. 25.
    Grotenhermen F (2003) Pharmacokinetics and pharmacodynamics of cannabinoids. Clin Pharmacokinet 42:327–360PubMedCrossRefGoogle Scholar
  26. 26.
    Friedman HS, Colvin OM, Skapek SX, Ludeman SM, Elion GB, Schold SC Jr, Jacobsen PF, Muhlbaier LH, Bigner DD (1988) Experimental chemotherapy of human medulloblastoma cell lines and transplantable xenografts with bifunctional alkylating agents. Cancer Res 48(15):4189–4195PubMedGoogle Scholar
  27. 27.
    Friedman HS, Colvin OM, Skapek SX, Ludeman SM, Elion GB, Schold SCJ, Jacobsen PF, Muhlbaier LH, Bigner DD (1988) Experimental chemotherapy of human medulloblastoma cell lines and transplantable xenografts with bifunctional alkylating agents. Cancer Res 48:4189–4195PubMedGoogle Scholar
  28. 28.
    Gehan EAA (1965) Generalized wilcoxon test for comparing arbitrarily singly-censored samples. Biometrika 52:203–223PubMedGoogle Scholar
  29. 29.
    Gowan SM, Hardcastle A, Hallsworth AE, Valenti MR, Hunter LJ, de Haven Brandon AK, Garrett MD, Raynaud F, Workman P, Aherne W, Eccles SA (2007) Application of meso scale technology for the measurement of phosphoproteins in human tumor xenografts. Assay Drug Dev Technol 5:391–401Google Scholar
  30. 30.
    Eccles SA, Massey A, Raynaud FI, Sharp SY, Box G, Valenti M, Patterson L, de Haven Brandon A, Gowan S, Boxall F, Aherne W, Rowlands M, Hayes A, Martins V, Urban F, Boxall K, Prodromou C, Pearl L, James K, Matthews TP, Cheung KM, Kalusa A, Jones K, McDonald E, Barril X, Brough PA, Cansfield JE, Dymock B, Drysdale MJ, Finch H, Howes R, Hubbard RE, Surgenor A, Webb P, Wood M, Wright L, Workman P (2008) NVP-AUY922: a novel heat shock protein 90 inhibitor active against xenograft tumor growth, angiogenesis, and metastasis. Cancer Res 68:2850–2860Google Scholar
  31. 31.
    Koul D (2008) PTEN signaling pathways in glioblastoma. Cancer Biol Ther 7:1321–1325PubMedCrossRefGoogle Scholar
  32. 32.
    Steck PA, Pershouse MA, Jasser SA, Yung WK, Lin H, Ligon AH, Langford LA, Baumgard ML, Hattier T, Davis T, Frye C, Hu R, Swedlund B, Teng DH, Tavtigian SV (1997) Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat Genet 15:356–362PubMedCrossRefGoogle Scholar
  33. 33.
    Sun MG, Williams J, Munoz-Pinedo C, Perkins GA, Brown JM, Ellisman MH, Green DR, Frey TG (2007) Correlated three-dimensional light and electron microscopy reveals transformation of mitochondria during apoptosis. Nat Cell Biol 9:1057–1065PubMedCrossRefGoogle Scholar
  34. 34.
    Jacobs VL, Valdes PA, Hickey WF, De Leo JA (2011) Current review of in vivo GBM rodent models: emphasis on the CNS-1 tumour model. ASN Neuro 3:171–181CrossRefGoogle Scholar
  35. 35.
    Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312PubMedCrossRefGoogle Scholar
  36. 36.
    Thornberry NA, Lazebnik Y (1998) Caspases: enemies within. Science 281:1312–1316PubMedCrossRefGoogle Scholar
  37. 37.
    Ellert-Miklaszewska A, Kaminska B, Konarska L (2005) Cannabinoids down-regulate PI3K/Akt and Erk signalling pathways and activate proapoptotic function of Bad protein. Cell Signal 17:25–37PubMedCrossRefGoogle Scholar
  38. 38.
    Greenhough A, Patsos HA, Williams AC, Paraskeva C (2007) The cannabinoid delta(9)-tetrahydrocannabinol inhibits RAS-MAPK and PI3K-AKT survival signalling and induces BAD-mediated apoptosis in colorectal cancer cells. Int J Cancer 121:2172–2180PubMedCrossRefGoogle Scholar
  39. 39.
    Jia W, Hegde VL, Singh NP, Sisco D, Grant S, Nagarkatti M, Nagarkatti PS (2006) Delta9-tetrahydrocannabinol-induced apoptosis in Jurkat leukemia T cells is regulated by translocation of Bad to mitochondria. Mol Cancer Res 4:549–562PubMedCrossRefGoogle Scholar
  40. 40.
    Cudaback E, Marrs W, Moeller T, Stella N (2010) The expression level of CB1 and CB2 receptors determines their efficacy at inducing apoptosis in astrocytomas. PLoS ONE 5(1):e8702PubMedCrossRefGoogle Scholar
  41. 41.
    Cattaneo E, Magrassi L, De-Fraja C, Conti L, Di Gennaro I, Butti G, Govoni S (1998) Variations in the levels of the JAK/STAT and ShcA proteins in human brain tumors. Anticancer Res 18:2381–2387PubMedGoogle Scholar
  42. 42.
    Caldera V, Mellai M, Annovazzi L, Valente G, Tessitore L, Schiffer D (2008) Stat3 expression and its correlation with proliferation and apoptosis/autophagy in gliomas. J Oncol Article ID 219241Google Scholar
  43. 43.
    Mizoguchi M, Betensky RA, Batchelor TT, Bernay DC, Louis DN, Nutt CL (2006) Activation of STAT3, MAPK, and AKT in malignant astrocytic gliomas: correlation with EGFR status, tumor grade, and survival. J Neuropathol Exp Neurol 65:1181–1188PubMedCrossRefGoogle Scholar
  44. 44.
    Liu Y, Li C, Lin J (2010) STAT3 as a therapeutic target for glioblastoma. Anticancer Agents Med Chem 10:512–519PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Steven N. Gurley
    • 1
  • Ammaar H. Abidi
    • 1
  • Patrick Allison
    • 1
  • Peihong Guan
    • 1
  • Christopher Duntsch
    • 2
  • Jon H. Robertson
    • 2
  • Stanley D. Kosanke
    • 3
  • Stephen T. Keir
    • 4
  • Darell D. Bigner
    • 5
  • Andrea J. Elberger
    • 6
  • Bob M. MooreII
    • 1
  1. 1.Department of Pharmaceutical SciencesThe University of Tennessee Health Science CenterMemphisUSA
  2. 2.Department of NeurosurgeryThe University of Tennessee Health Science CenterMemphisUSA
  3. 3.Department of PathologyUniversity of Oklahoma Health Sciences CenterOklahoma CityUSA
  4. 4.Department of SurgeryDuke UniversityDurhamUSA
  5. 5.Department of PathologyDuke UniversityDurhamUSA
  6. 6.Department of Anatomy and NeurobiologyThe University of Tennessee Health Science CenterMemphisUSA

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