Skip to main content

Advertisement

SpringerLink
Log in

Palmitoyl Ascorbate Liposomes and Free Ascorbic Acid: Comparison of Anticancer Therapeutic Effects Upon Parenteral Administration

  • Research Paper
  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

ABSTRACT

Purpose

To evaluate and compare anticancer therapeutic effect of palmitoyl ascorbate liposomes (PAL) and free ascorbic acid (AA).

Methods

Liposomes incorporating palmitoyl ascorbate (PA) were prepared and evaluated for PA content by HPLC. To elucidate mechanism of action of cell death in vitro, effect of various H2O2 scavengers and metal chelators on PA-mediated cytotoxicity was studied. Effect of various combinations of PAL and free AA on in vitro cytotoxicity was evaluated on 4T1 cells. In vivo, PAL formulation was modified with polyethylene glycol; effect of PEGylation on in vitro cytotoxicity was evaluated. Biodistribution of PEG-PAL formulation was investigated in female Balb/c mice bearing murine mammary carcinoma (4T1 cells). In vivo anticancer activity of PEG-PAL (PEG-PAL equivalent to 20 mg/kg of PA injected intravenously on alternate days) was compared with free AA therapy in same model.

Results

PEG-PAL treatment was significantly more effective than free AA treatment in slowing tumor growth.

Conclusions

Nanoparticle formulations incorporating PA can kill cancer cells in vitro. The mechanism of PA cytotoxicity is based on production of extracellular reactive oxygen species and involves intracellular transition metals.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

AA:

ascorbic acid

DHAA:

dehydroascorbic acid

DFO:

desferrioxamine mesylate

DMEM:

Dulbecco’s Modified Eagle Medium

DTPA-PE:

1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-diethylenetriaminepentaacetic acid

EDTA:

ethylenediaminetetraacetic acid

FBS:

fetal bovine serum

GLUTs:

glucose transporters

HIF:

hypoxia-inducible factors

PA:

palmitoyl ascorbate

PAL:

palmitoyl ascorbate liposomes

PEG2000-PE:

1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (poly (ethylene glycol))-2000] (PEG2000-PE)

PL:

plain liposomes

ROS:

reactive oxygen species

SOD:

superoxide dismutase

TCEP:

tris(2-carboxyethyl) phosphine hydrochloride

REFERENCES

  1. Chen Q, Espey MG, Sun AY, Pooput C, Kirk KL, Krishna MC, et al. Pharmacologic doses of ascorbate act as a prooxidant and decrease growth of aggressive tumor xenografts in mice. Proc Natl Acad Sci USA. 2008;105(32):11105–9.

    Article  PubMed  CAS  Google Scholar 

  2. Chen Q, Espey MG, Sun AY, Lee JH, Krishna MC, Shacter E, et al. Ascorbate in pharmacologic concentrations selectively generates ascorbate radical and hydrogen peroxide in extracellular fluid in vivo. Proc Natl Acad Sci USA. 2007;104(21):8749–54.

    Article  PubMed  CAS  Google Scholar 

  3. Belin S, Kaya F, Duisit G, Giacometti S, Ciccolini J, Fontes M. Antiproliferative effect of ascorbic acid is associated with the inhibition of genes necessary to cell cycle progression. PLoS One. 2009;4(2):e4409.

    Article  PubMed  Google Scholar 

  4. Verrax J, Calderon PB. Pharmacologic concentrations of ascorbate are achieved by parenteral administration and exhibit antitumoral effects. Free Radic Biol Med. 2009;47(1):32–40.

    Article  PubMed  CAS  Google Scholar 

  5. Hoffer LJ, Levine M, Assouline S, Melnychuk D, Padayatty SJ, Rosadiuk K, et al. clinical trial of i.v. ascorbic acid in advanced malignancy. Ann Oncol. 2008;19(11):1969–74.

    Article  PubMed  CAS  Google Scholar 

  6. Cameron E, Pauling L. Supplemental ascorbate in the supportive treatment of cancer: prolongation of survival times in terminal human cancer. Proc Natl Acad Sci USA. 1976;73(10):3685–9.

    Article  PubMed  CAS  Google Scholar 

  7. Cameron E, Pauling L. Supplemental ascorbate in the supportive treatment of cancer: reevaluation of prolongation of survival times in terminal human cancer. Proc Natl Acad Sci USA. 1978;75(9):4538–42.

    Article  PubMed  CAS  Google Scholar 

  8. Yeom CH, Jung GC, Song KJ. Changes of terminal cancer patients’ health-related quality of life after high dose vitamin C administration. J Korean Med Sci. 2007;22(1):7–11.

    Article  PubMed  CAS  Google Scholar 

  9. Torchilin VP. Targeted pharmaceutical nanocarriers for cancer therapy and imaging. AAPS J. 2007;9(2):E128–47.

    Article  PubMed  CAS  Google Scholar 

  10. Miwa N, Yamazaki H, Nagaoka Y, Kageyama K, Onoyama Y, Matsui-Yuasa I, et al. Altered production of the active oxygen species is involved in enhanced cytotoxic action of acylated derivatives of ascorbate to tumor cells. Biochim Biophys Acta. 1988;972(2):144–51.

    Article  PubMed  CAS  Google Scholar 

  11. D’Souza GG, Wang T, Rockwell K, Torchilin VP. Surface modification of pharmaceutical nanocarriers with ascorbate residues improves their tumor-cell association and killing and the cytotoxic action of encapsulated paclitaxel in vitro. Pharm Res. 2008;25(11):2567–72.

    Article  PubMed  Google Scholar 

  12. Sawant RR, Vaze OS, Rockwell K, Torchilin VP. Palmitoyl ascorbate-modified liposomes as nanoparticle platform for ascorbate-mediated cytotoxicity and paclitaxel co-delivery. Eur J Pharm Biopharm. 2010;75(3):321–6.

    Article  PubMed  CAS  Google Scholar 

  13. Agus DB, Vera JC, Golde DW. Stromal cell oxidation: a mechanism by which tumors obtain vitamin C. Cancer Res. 1999;59(18):4555–8.

    PubMed  CAS  Google Scholar 

  14. Agus DB, Gambhir SS, Pardridge WM, Spielholz C, Baselga J, Vera JC, et al. Vitamin C crosses the blood-brain barrier in the oxidized form through the glucose transporters. J Clin Invest. 1997;100(11):2842–8.

    Article  PubMed  CAS  Google Scholar 

  15. Rumsey SC, Kwon O, Xu GW, Burant CF, Simpson I, Levine M. Glucose transporter isoforms GLUT1 and GLUT3 transport dehydroascorbic acid. J Biol Chem. 1997;272(30):18982–9.

    Article  PubMed  CAS  Google Scholar 

  16. Rumsey SC, Daruwala R, Al-Hasani H, Zarnowski MJ, Simpson IA, Levine M. Dehydroascorbic acid transport by GLUT4 in Xenopus oocytes and isolated rat adipocytes. J Biol Chem. 2000;275(36):28246–53.

    PubMed  CAS  Google Scholar 

  17. Dai J, Weinberg RS, Waxman S, Jing Y. Malignant cells can be sensitized to undergo growth inhibition and apoptosis by arsenic trioxide through modulation of the glutathione redox system. Blood. 1999;93(1):268–77.

    PubMed  CAS  Google Scholar 

  18. Lu H, Dalgard CL, Mohyeldin A, McFate T, Tait AS, Verma A. Reversible inactivation of HIF-1 prolyl hydroxylases allows cell metabolism to control basal HIF-1. J Biol Chem. 2005;280(51):41928–39.

    Article  PubMed  CAS  Google Scholar 

  19. Pan Y, Mansfield KD, Bertozzi CC, Rudenko V, Chan DA, Giaccia AJ, et al. Multiple factors affecting cellular redox status and energy metabolism modulate hypoxia-inducible factor prolyl hydroxylase activity in vivo and in vitro. Mol Cell Biol. 2007;27(3):912–25.

    Article  PubMed  CAS  Google Scholar 

  20. Knowles HJ, Raval RR, Harris AL, Ratcliffe PJ. Effect of ascorbate on the activity of hypoxia-inducible factor in cancer cells. Cancer Res. 2003;63(8):1764–8.

    PubMed  CAS  Google Scholar 

  21. Vissers MC, Gunningham SP, Morrison MJ, Dachs GU, Currie MJ. Modulation of hypoxia-inducible factor-1 alpha in cultured primary cells by intracellular ascorbate. Free Radic Biol Med. 2007;42(6):765–72.

    Article  PubMed  CAS  Google Scholar 

  22. Knowles HJ, Mole DR, Ratcliffe PJ, Harris AL. Normoxic stabilization of hypoxia-inducible factor-1alpha by modulation of the labile iron pool in differentiating U937 macrophages: effect of natural resistance-associated macrophage protein 1. Cancer Res. 2006;66(5):2600–7.

    Article  PubMed  CAS  Google Scholar 

  23. Chen Q, Espey MG, Krishna MC, Mitchell JB, Corpe CP, Buettner GR, et al. Pharmacologic ascorbic acid concentrations selectively kill cancer cells: action as a pro-drug to deliver hydrogen peroxide to tissues. Proc Natl Acad Sci USA. 2005;102(38):13604–9.

    Article  PubMed  CAS  Google Scholar 

  24. Rosenblat G, Graham MF, Jonas A, Tarshis M, Schubert SY, Tabak M, et al. Effect of ascorbic acid and its hydrophobic derivative palmitoyl ascorbate on the redox state of primary human fibroblasts. J Med Food. 2001;4(2):107–15.

    Article  PubMed  CAS  Google Scholar 

  25. Yuan F, Leunig M, Huang SK, Berk DA, Papahadjopoulos D, Jain RK. Microvascular permeability and interstitial penetration of sterically stabilized (stealth) liposomes in a human tumor xenograft. Cancer Res. 1994;54(13):3352–6.

    PubMed  CAS  Google Scholar 

  26. Padayatty SJ, Sun H, Wang Y, Riordan HD, Hewitt SM, Katz A, et al. Vitamin C pharmacokinetics: implications for oral and intravenous use. Ann Intern Med. 2004;140(7):533–7.

    PubMed  CAS  Google Scholar 

  27. Levine M, Conry-Cantilena C, Wang Y, Welch RW, Washko PW, Dhariwal KR, et al. Vitamin C pharmacokinetics in healthy volunteers: evidence for a recommended dietary allowance. Proc Natl Acad Sci USA. 1996;93(8):3704–9.

    Article  PubMed  CAS  Google Scholar 

  28. Torchilin VP. Recent advances with liposomes as pharmaceutical carriers. Nat Rev Drug Discov. 2005;4(2):145–60.

    Article  PubMed  CAS  Google Scholar 

  29. Hashizume H, Baluk P, Morikawa S, McLean JW, Thurston G, Roberge S, et al. Openings between defective endothelial cells explain tumor vessel leakiness. Am J Pathol. 2000;156(4):1363–80.

    Article  PubMed  CAS  Google Scholar 

  30. Maeda H, Wu J, Sawa T, Matsumura Y, Hori K. Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. J Control Release. 2000;65(1–2):271–84.

    Article  PubMed  CAS  Google Scholar 

  31. Bartlett DW, Su H, Hildebrandt IJ, Weber WA, Davis ME. Impact of tumor-specific targeting on the biodistribution and efficacy of siRNA nanoparticles measured by multimodality in vivo imaging. Proc Natl Acad Sci USA. 2007;104(39):15549–54.

    Article  PubMed  CAS  Google Scholar 

  32. Kirpotin DB, Drummond DC, Shao Y, Shalaby MR, Hong K, Nielsen UB, et al. Antibody targeting of long-circulating lipidic nanoparticles does not increase tumor localization but does increase internalization in animal models. Cancer Res. 2006;66(13):6732–40.

    Article  PubMed  CAS  Google Scholar 

  33. Kurbacher CM, Wagner U, Kolster B, Andreotti PE, Krebs D, Bruckner HW. Ascorbic acid (vitamin C) improves the antineoplastic activity of doxorubicin, cisplatin, and paclitaxel in human breast carcinoma cells in vitro. Cancer Lett. 1996;103(2):183–9.

    Article  PubMed  CAS  Google Scholar 

  34. Bahlis NJ, McCafferty-Grad J, Jordan-McMurry I, Neil J, Reis I, Kharfan-Dabaja M, et al. Feasibility and correlates of arsenic trioxide combined with ascorbic acid-mediated depletion of intracellular glutathione for the treatment of relapsed/refractory multiple myeloma. Clin Cancer Res. 2002;8(12):3658–68.

    PubMed  CAS  Google Scholar 

Download references

ACKNOWLEDGMENTS & DISCLOSURES

This research is based on a hypothesis originated and proposed by Anthony R. Manganaro. Funding was provided by Anthony R. Manganaro.

Author information

Author notes

  1. Gerard G. M. D’Souza

    Present address: Department of Pharmaceutical Sciences, Massachusetts College of Pharmacy and Health Sciences, 179 Longwood Avenue, Boston, Massachusetts, 02115, USA

Authors and Affiliations

  1. Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, Mugar Building, Room 312, 360 Huntington Avenue, Boston, Massachusetts, 02115, USA

    Rupa R. Sawant, Onkar S. Vaze, Tao Wang, Gerard G. M. D’Souza, Karen Rockwell, Keyur Gada, Ban-An Khaw & Vladimir P. Torchilin

Authors
  1. Rupa R. Sawant
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Onkar S. Vaze
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gerard G. M. D’Souza
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Karen Rockwell
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Keyur Gada
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ban-An Khaw
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Vladimir P. Torchilin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vladimir P. Torchilin.

Additional information

Rupa R. Sawant and Onkar S. Vaze contributed equally to this study.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sawant, R.R., Vaze, O.S., Wang, T. et al. Palmitoyl Ascorbate Liposomes and Free Ascorbic Acid: Comparison of Anticancer Therapeutic Effects Upon Parenteral Administration. Pharm Res 29, 375–383 (2012). https://doi.org/10.1007/s11095-011-0557-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-011-0557-8

KEY WORDS

Search

Navigation