Advertisement

AAPS PharmSciTech

, Volume 19, Issue 5, pp 2133–2143 | Cite as

Ceramide-Fabricated Co-Loaded Liposomes for the Synergistic Treatment of Hepatocellular Carcinoma

  • Xiaolan Yin
  • Yanan Xiao
  • Leiqiang Han
  • Bo Zhang
  • Tianqi Wang
  • Zhihui Su
  • Na Zhang
Research Article
  • 80 Downloads

Abstract

Combination therapy is one of the important methods to improve therapeutic effect on the treatment of hepatocellular carcinoma (HCC). Sorafenib (SF) is a canonical US Food and Drug Administration-approved multikinase molecule inhibitor against HCC. However, therapeutic benefit with Sorafenib alone was usually unsatisfactory. Ceramide (CE) is an endogenous bioactive sphingolipid, which has a strong potential to suppress various tumors. The combination of SF and CE was hoping to exert maximum synergistic antitumor effect through different tumor-suppressible mechanisms. In this respect, SF and CE co-loaded liposomes (SF/CE-liposomes) were developed to verify synergistic antitumor efficacy. The optimal molar ratio of SF and CE was determined through combination index. SF/CE-liposomes were prepared by thin-film hydration method, which exhibited spherical or ellipsoidal shape. Particle size of SF/CE-liposomes was 174 ± 4 nm with homogeneous distribution. Release profile of SF demonstrated that addition of CE imposed no significant impact on the release of SF. SF/CE-liposomes exhibited acceptable stability in different media and desirable storage stability over 30 days at 4°C. In vitro cellular uptake confirmed that SF/CE-liposomes could be efficiently internalized into HepG2 cells. In vitro cytotoxicity evaluation indicated that SF/CE-liposomes exhibited higher cytotoxicity on HepG2 cells. IC50 value of SF/CE-liposomes was 11.5 ± 0.44 μM, which was significantly lower than that of SF-liposomes (**p < 0.01). Evaluation of in vivo synergistic effect on H22-bearing mice verified that SF/CE-liposomes achieved robust antitumor activity in preventing tumor growth. All results suggested that SF/CE-liposomes might be served as an efficient co-delivery system for improving therapeutic efficacy of HCC.

KEY WORDS

hepatocellular carcinoma co-delivery sorafenib ceramide liposomes 

Notes

Acknowledgments

We are very thankful for Dr. Livesey David Olerile (School of Pharmaceutical Sciences, Shandong University) for modifying the English language of this manuscript.

Funding Information

This work was partially funded by the National Natural Science Foundation of China (no. 81573368) and the Science and Technology Development Project of Shandong Province (2014GGE27121).

Compliance with Ethical Standards

All of the animal experiments were performed with the approval of Institutional  Animal Care and Use Committee of Shandong University and in compliance with the Animal Management Rules of Ministry of Health of the People's Republic of China (document number 55, 2001).

Disclosure

The authors declare that they have no conflicts of interest in this work.

References

  1. 1.
    Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology. 2006;24(14):2137–50.CrossRefGoogle Scholar
  2. 2.
    Bruix J, Sherman M, American Association for the Study of Liver D. Management of hepatocellular carcinoma: an update. Hepatology. 2011;53(3):1020–2.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Bruix J, Gores GJ, Mazzaferro V. Hepatocellular carcinoma: clinical frontiers and perspectives. Gut. 2014;63(5):844–55.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Hsu CH, Shen YC, Shao YY, Hsu C, Cheng AL. Sorafenib in advanced hepatocellular carcinoma: current status and future perspectives. Journal of Hepatocellular Carcinoma. 2014;1(1):85–99.PubMedPubMedCentralGoogle Scholar
  5. 5.
    Ogasawara S, Chiba T, Ooka Y, Suzuki E, Inoue M, Wakamatsu T, et al. Analysis of Sorafenib outcome: focusing on the clinical course in patients with hepatocellular carcinoma. PLoS One. 2016;11(8):e0161303.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Thillai K, Ross P, Sarker D. Molecularly targeted therapy for advanced hepatocellular carcinoma—a drug development crisis? World Journal of Gastrointestinal Oncology. 2016;8(2):173–85.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Huang J, Tang Q, Wang C, Yu H, Feng Z, Zhu J. Molecularly targeted therapy of human hepatocellular carcinoma xenografts with radio-iodinated anti-VEGFR2 murine-human chimeric Fab. Sci Rep. 2015;5:10660.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, et al. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 2008;359(4):378–90.CrossRefPubMedGoogle Scholar
  9. 9.
    Cheng AL, Kang YK, Chen Z, Tsao CJ, Qin S, Kim JS, et al. Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma: a phase III randomised, double-blind, placebo-controlled trial. The Lancet Oncology. 2009;10(1):25–34.CrossRefPubMedGoogle Scholar
  10. 10.
    Small R, Lubezky N, Ben-Haim M. Current controversies in the surgical management of colorectal cancer metastases to the liver. The Israel Medical Association Journal: IMAJ. 2007;9(10):742–7.PubMedGoogle Scholar
  11. 11.
    Kemp JA, Shim MS, Heo CY, Kwon YJ. “Combo” nanomedicine: co-delivery of multi-modal therapeutics for efficient, targeted, and safe cancer therapy. Adv Drug Deliv Rev. 2016;98:3–18.CrossRefPubMedGoogle Scholar
  12. 12.
    Galuppo R, Ramaiah D, Ponte OM, Gedaly R. Molecular therapies in hepatocellular carcinoma: what can we target? Dig Dis Sci. 2014;59(8):1688–97.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Huynh H. Molecularly targeted therapy in hepatocellular carcinoma. Biochem Pharmacol. 2010;80(5):550–60.CrossRefPubMedGoogle Scholar
  14. 14.
    Richly H, Kupsch P, Passage K, Grubert M, Hilger RA, Voigtmann R, et al. Results of a phase I trial of BAY 43-9006 in combination with doxorubicin in patients with primary hepatic cancer. Int J Clin Pharmacol Ther. 2004;42(11):650–1.CrossRefPubMedGoogle Scholar
  15. 15.
    Abou-Alfa GK, Johnson P, Knox JJ, Capanu M, Davidenko I, Lacava J, et al. Doxorubicin plus sorafenib vs doxorubicin alone in patients with advanced hepatocellular carcinoma: a randomized trial. JAMA. 2010;304(19):2154–60.CrossRefPubMedGoogle Scholar
  16. 16.
    Woo HY, Heo J. Sorafenib in liver cancer. Expert Opin Pharmacother. 2012;13(7):1059–67.CrossRefPubMedGoogle Scholar
  17. 17.
    Huynh H, Ngo VC, Koong HN, Poon D, Choo SP, Thng CH, et al. Sorafenib and rapamycin induce growth suppression in mouse models of hepatocellular carcinoma. J Cell Mol Med. 2009;13(8B):2673–83.CrossRefPubMedGoogle Scholar
  18. 18.
    Morad SA, Cabot MC. Ceramide-orchestrated signalling in cancer cells. Nat Rev Cancer. 2013;13(1):51–65.CrossRefPubMedGoogle Scholar
  19. 19.
    Dany M, Ogretmen B. Ceramide induced mitophagy and tumor suppression. Biochim Biophys Acta. 2015;1853(10):2834–45.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Zhang P, Fu C, Hu Y, Dong C, Song Y, Song E. C6-ceramide nanoliposome suppresses tumor metastasis by eliciting PI3K and PKCzeta tumor-suppressive activities and regulating integrin affinity modulation. Sci Rep. 2015;5:9275.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Li F, Zhang N. Ceramide: therapeutic potential in combination therapy for cancer treatment. Curr Drug Metab. 2015;17(1):37–51.CrossRefPubMedGoogle Scholar
  22. 22.
    Overbye A, Holsaeter AM, Markus F, Skalko-Basnet N, Iversen TG, Torgersen ML, et al. Ceramide-containing liposomes with doxorubicin: time and cell-dependent effect of C6 and C12 ceramide. Oncotarget. 2017;8(44):76921–34.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Carvalho VFM, Migotto A, Giacone DV, de Lemos DP, Zanoni TB, Maria-Engler SS, et al. Co-encapsulation of paclitaxel and C6 ceramide in tributyrin-containing nanocarriers improve co-localization in the skin and potentiate cytotoxic effects in 2D and 3D models. Eur J Pharm Sci. 2017;109:131–43.CrossRefPubMedGoogle Scholar
  24. 24.
    Wang T, Feng L, Yang S, Liu Y, Zhang N. Ceramide lipid-based nanosuspension for enhanced delivery of docetaxel with synergistic antitumor efficiency. Drug Delivery. 2017;24(1):800–10.CrossRefPubMedGoogle Scholar
  25. 25.
    Fonseca NA, Gomes-da-Silva LC, Moura V, Simoes S, Moreira JN. Simultaneous active intracellular delivery of doxorubicin and C6-ceramide shifts the additive/antagonistic drug interaction of non-encapsulated combination. Journal of Controlled Release: Official Journal of the Controlled Release Society. 2014;196:122–31.CrossRefGoogle Scholar
  26. 26.
    Wang Y, Ding Y, Liu Z, Liu X, Chen L, Yan W. Bioactive lipids-based pH sensitive micelles for co-delivery of doxorubicin and ceramide to overcome multidrug resistance in leukemia. Pharm Res. 2013;30(11):2902–16.CrossRefPubMedGoogle Scholar
  27. 27.
    Yang L, Zheng LY, Tian Y, Zhang ZQ, Dong WL, Wang XF, et al. C6 ceramide dramatically enhances docetaxel-induced growth inhibition and apoptosis in cultured breast cancer cells: a mechanism study. Exp Cell Res. 2015;332(1):47–59.CrossRefPubMedGoogle Scholar
  28. 28.
    Feng LX, Li M, Liu YJ, Yang SM, Zhang N. Synergistic enhancement of cancer therapy using a combination of Ceramide and Docetaxel. Int J Mol Sci. 2014;15(3):4201–20.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Sriraman SK, Pan J, Sarisozen C, Luther E, Torchilin V. Enhanced cytotoxicity of folic acid-targeted liposomes co-loaded with C6 ceramide and doxorubicin: in vitro evaluation on HeLa, A2780-ADR, and H69-AR cells. Mol Pharm. 2016;13(2):428–37.CrossRefPubMedGoogle Scholar
  30. 30.
    Rivera IG, Ordonez M, Presa N, Gangoiti P, Gomez-Larrauri A, Trueba M, et al. Ceramide 1-phosphate regulates cell migration and invasion of human pancreatic cancer cells. Biochem Pharmacol. 2016;102:107–19.CrossRefPubMedGoogle Scholar
  31. 31.
    Barth BM, Cabot MC, Kester M. Ceramide-based therapeutics for the treatment of cancer. Anti Cancer Agents Med Chem. 2011;11(9):911–9.CrossRefGoogle Scholar
  32. 32.
    Stover T, Kester M. Liposomal delivery enhances short-chain ceramide-induced apoptosis of breast cancer cells. J Pharmacol Exp Ther. 2003;307(2):468–75.CrossRefPubMedGoogle Scholar
  33. 33.
    Tagaram HR, Divittore NA, Barth BM, Kaiser JM, Avella D, Kimchi ET, et al. Nanoliposomal ceramide prevents in vivo growth of hepatocellular carcinoma. Gut. 2011;60(5):695–701.CrossRefPubMedGoogle Scholar
  34. 34.
    Liu M, Gu P, Guo W, Fan X. C6 ceramide sensitizes the anti-hepatocellular carcinoma (HCC) activity by AZD-8055, a novel mTORC1/2 dual inhibitor. Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine. 2016;37(8):11039–48.CrossRefGoogle Scholar
  35. 35.
    Kester M, Bassler J, Fox TE, Carter CJ, Davidson JA, Parette MR. Preclinical development of a C6-ceramide nanoliposome, a novel sphingolipid therapeutic. Biol Chem. 2015;396(6–7):737–47.PubMedGoogle Scholar
  36. 36.
    Jiang S, Wang Q, Feng M, Li J, Guan Z, An D, et al. C2-ceramide enhances sorafenib-induced caspase-dependent apoptosis via PI3K/AKT/mTOR and Erk signaling pathways in HCC cells. Appl Microbiol Biotechnol. 2017;101(4):1535–46.CrossRefPubMedGoogle Scholar
  37. 37.
    Lim SB, Banerjee A, Onyuksel H. Improvement of drug safety by the use of lipid-based nanocarriers. J Control Release. 2012;163(1):34–45.CrossRefPubMedGoogle Scholar
  38. 38.
    Yang F, Jin C, Jiang Y, Li J, Di Y, Ni Q, et al. Liposome based delivery systems in pancreatic cancer treatment: from bench to bedside. Cancer Treat Rev. 2011;37(8):633–42.CrossRefPubMedGoogle Scholar
  39. 39.
    Allen TM, Cullis PR. Liposomal drug delivery systems: from concept to clinical applications. Adv Drug Deliv Rev. 2013;65(1):36–48.CrossRefPubMedGoogle Scholar
  40. 40.
    Damitz R, Chauhan A. Parenteral emulsions and liposomes to treat drug overdose. Adv Drug Deliv Rev. 2015;90:12–23.CrossRefPubMedGoogle Scholar
  41. 41.
    Iwabuchi K, Nakayama H, Oizumi A, Suga Y, Ogawa H, Takamori K. Role of Ceramide from glycosphingolipids and its metabolites in immunological and inflammatory responses in humans. Mediat Inflamm. 2015;2015(10):120748.Google Scholar
  42. 42.
    Castro BM, Prieto M, Silva LC. Ceramide: a simple sphingolipid with unique biophysical properties. Prog Lipid Res. 2014;54(1):53–67.CrossRefPubMedGoogle Scholar
  43. 43.
    Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzym Regul. 1984;22(84):27–55.CrossRefGoogle Scholar
  44. 44.
    Dhule SS, Penfornis P, He JB, Harris MR, Terry T, John V, et al. The combined effect of encapsulating curcumin and C6 ceramide in liposomal nanoparticles against osteosarcoma. Mol Pharm. 2014;11(2):417–27.CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Menon P, Yin TY, Misran M. Preparation and characterization of liposomes coated with DEAE-dextran. Colloid Surface A. 2015;481(7):345–50.CrossRefGoogle Scholar
  46. 46.
    Xiao Y, Liu Y, Yang S, Zhang B, Wang T, Jiang D, et al. Sorafenib and gadolinium co-loaded liposomes for drug delivery and MRI-guided HCC treatment. Colloids Surf B: Biointerfaces. 2016;141:83–92.CrossRefPubMedGoogle Scholar
  47. 47.
    Suk JS, Xu QG, Kim N, Hanes J, Ensign LM. PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev. 2016;99:28–51.CrossRefPubMedGoogle Scholar
  48. 48.
    Yang SM, Zhang B, Gong XW, Wang TQ, Liu YJ, Zhang N. In vivo biodistribution, biocompatibility, and efficacy of sorafenib-loaded lipid-based nanosuspensions evaluated experimentally in cancer. Int J Nanomedicine. 2016;11:2329–43.PubMedPubMedCentralGoogle Scholar
  49. 49.
    Matsumura Y, Maeda H. A new concept for macromolecular therapeutics in cancer-chemotherapy—mechanism of tumoritropic accumulation of proteins and the antitumor agent Smancs. Cancer Res. 1986;46(12):6387–92.PubMedGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Xiaolan Yin
    • 1
  • Yanan Xiao
    • 1
  • Leiqiang Han
    • 1
  • Bo Zhang
    • 1
  • Tianqi Wang
    • 1
  • Zhihui Su
    • 1
  • Na Zhang
    • 1
  1. 1.School of Pharmaceutical Sciences, Key Laboratory of Chemical Biology (Ministry of Education)Shandong UniversityJi’nanPeople’s Republic of China

Personalised recommendations