Skip to main content
PMC home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1996 Jul;74(1):43–48. doi: 10.1038/bjc.1996.313

An improved method of encapsulation of doxorubicin in liposomes: pharmacological, toxicological and therapeutic evaluation.

P C Gokhale 1, B Radhakrishnan 1, S R Husain 1, D R Abernethy 1, R Sacher 1, A Dritschilo 1, A Rahman 1
PMCID: PMC2074597  PMID: 8679456

Abstract

We describe here an improved method of encapsulating doxorubicin in liposomes using phosphatidylcholine, cholesterol and synthetic tetramyristoyl cardiolipin. With this new composition of lipids the entrapment of doxorubicin was found to be > 90%. Cytotoxicity studies using vincristine-resistant HL-60/VCR leukaemia cells showed that liposome-encapsulated doxorubicin reverses multidrug resistance 5-fold compared with conventional doxorubicin and at levels equivalent to that obtained using liposomes with natural cardiolipin. In normal mice, liposome-encapsulated doxorubicin was much less toxic than the conventional drug. A dose of 25 mg kg-1 i.v. of conventional doxorubicin produced 100% mortality in mice by day 14, whereas liposomal doxorubicin exhibited only 10% mortality by day 60. Liposomal doxorubicin demonstrated enhanced anti-tumour activity against murine ascitic L1210 leukaemia compared with conventional doxorubicin. At a dose of 15 mg kg-1, liposomal doxorubicin increased the median life span with 12 of 18 long-term (60 days) survivors compared with only 3 of 18 with conventional drug. Mice injected i.v. with liposomal doxorubicin had plasma levels 44-fold higher than conventional doxorubicin, producing significantly higher (P < 0.02) area under the plasma concentration curve. An altered tissue distribution was also observed with liposomal doxorubicin; cardiac tissue demonstrating at least 2-fold lower levels with liposomal doxorubicin probably accounting for its lower toxicity. This altered pharmacokinetics of liposome-encapsulated doxorubicin, providing enhanced therapeutic advantage and the ability to modulate multidrug resistance, could be useful in a clinical setting.

Full text

PDF
47

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Biedler J. L., Riehm H. Cellular resistance to actinomycin D in Chinese hamster cells in vitro: cross-resistance, radioautographic, and cytogenetic studies. Cancer Res. 1970 Apr;30(4):1174–1184. [PubMed] [Google Scholar]
  2. Dano K. Active outward transport of daunomycin in resistant Ehrlich ascites tumor cells. Biochim Biophys Acta. 1973 Oct 25;323(3):466–483. doi: 10.1016/0005-2736(73)90191-0. [DOI] [PubMed] [Google Scholar]
  3. Delgado G., Potkul R. K., Treat J. A., Lewandowski G. S., Barter J. F., Forst D., Rahman A. A phase I/II study of intraperitoneally administered doxorubicin entrapped in cardiolipin liposomes in patients with ovarian cancer. Am J Obstet Gynecol. 1989 Apr;160(4):812–819. doi: 10.1016/0002-9378(89)90296-2. [DOI] [PubMed] [Google Scholar]
  4. Forssen E. A., Tökès Z. A. Use of anionic liposomes for the reduction of chronic doxorubicin-induced cardiotoxicity. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1873–1877. doi: 10.1073/pnas.78.3.1873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gabizon A., Dagan A., Goren D., Barenholz Y., Fuks Z. Liposomes as in vivo carriers of adriamycin: reduced cardiac uptake and preserved antitumor activity in mice. Cancer Res. 1982 Nov;42(11):4734–4739. [PubMed] [Google Scholar]
  6. Goormaghtigh E., Ruysschaert J. M. Anthracycline glycoside-membrane interactions. Biochim Biophys Acta. 1984 Sep 3;779(3):271–288. doi: 10.1016/0304-4157(84)90013-3. [DOI] [PubMed] [Google Scholar]
  7. Gottesman M. M., Pastan I. The multidrug transporter, a double-edged sword. J Biol Chem. 1988 Sep 5;263(25):12163–12166. [PubMed] [Google Scholar]
  8. Gregoriadis G., Wills E. J., Swain C. P., Tavill A. S. Drug-carrier potential of liposomes in cancer chemotherapy. Lancet. 1974 Jun 29;1(7870):1313–1316. doi: 10.1016/s0140-6736(74)90682-5. [DOI] [PubMed] [Google Scholar]
  9. Herman E. H., Rahman A., Ferrans V. J., Vick J. A., Schein P. S. Prevention of chronic doxorubicin cardiotoxicity in beagles by liposomal encapsulation. Cancer Res. 1983 Nov;43(11):5427–5432. [PubMed] [Google Scholar]
  10. Horio M., Gottesman M. M., Pastan I. ATP-dependent transport of vinblastine in vesicles from human multidrug-resistant cells. Proc Natl Acad Sci U S A. 1988 May;85(10):3580–3584. doi: 10.1073/pnas.85.10.3580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Juliano R. L., Ling V. A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta. 1976 Nov 11;455(1):152–162. doi: 10.1016/0005-2736(76)90160-7. [DOI] [PubMed] [Google Scholar]
  12. Kartner N., Evernden-Porelle D., Bradley G., Ling V. Detection of P-glycoprotein in multidrug-resistant cell lines by monoclonal antibodies. 1985 Aug 29-Sep 4Nature. 316(6031):820–823. doi: 10.1038/316820a0. [DOI] [PubMed] [Google Scholar]
  13. Kim S. Liposomes as carriers of cancer chemotherapy. Current status and future prospects. Drugs. 1993 Oct;46(4):618–638. doi: 10.2165/00003495-199346040-00004. [DOI] [PubMed] [Google Scholar]
  14. McGrath T., Center M. S. Mechanisms of multidrug resistance in HL60 cells: evidence that a surface membrane protein distinct from P-glycoprotein contributes to reduced cellular accumulation of drug. Cancer Res. 1988 Jul 15;48(14):3959–3963. [PubMed] [Google Scholar]
  15. Mickisch G. H., Rahman A., Pastan I., Gottesman M. M. Increased effectiveness of liposome-encapsulated doxorubicin in multidrug-resistant-transgenic mice compared with free doxorubicin. J Natl Cancer Inst. 1992 May 20;84(10):804–805. doi: 10.1093/jnci/84.10.804. [DOI] [PubMed] [Google Scholar]
  16. Minow R. A., Benjamin R. S., Lee E. T., Gottlieb J. A. Adriamycin cardiomyopathy--risk factors. Cancer. 1977 Apr;39(4):1397–1402. doi: 10.1002/1097-0142(197704)39:4<1397::aid-cncr2820390407>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  17. O'Bryan R. M., Baker L. H., Gottlieb J. E., Rivkin S. E., Balcerzak S. P., Grumet G. N., Salmon S. E., Moon T. E., Hoogstraten B. Dose response evaluation of adriamycin in human neoplasia. Cancer. 1977 May;39(5):1940–1948. doi: 10.1002/1097-0142(197705)39:5<1940::aid-cncr2820390505>3.0.co;2-0. [DOI] [PubMed] [Google Scholar]
  18. Olson F., Mayhew E., Maslow D., Rustum Y., Szoka F. Characterization, toxicity and therapeutic efficacy of adriamycin encapsulated in liposomes. Eur J Cancer Clin Oncol. 1982 Feb;18(2):167–176. doi: 10.1016/0277-5379(82)90060-8. [DOI] [PubMed] [Google Scholar]
  19. Oudard S., Thierry A., Jorgensen T. J., Rahman A. Sensitization of multidrug-resistant colon cancer cells to doxorubicin encapsulated in liposomes. Cancer Chemother Pharmacol. 1991;28(4):259–265. doi: 10.1007/BF00685532. [DOI] [PubMed] [Google Scholar]
  20. Rahman A., Fumagalli A., Barbieri B., Schein P. S., Casazza A. M. Antitumor and toxicity evaluation of free doxorubicin and doxorubicin entrapped in cardiolipin liposomes. Cancer Chemother Pharmacol. 1986;16(1):22–27. doi: 10.1007/BF00255281. [DOI] [PubMed] [Google Scholar]
  21. Rahman A., Ganjei A., Neefe J. R. Comparative immunotoxicity of free doxorubicin and doxorubicin encapsulated in cardiolipin liposomes. Cancer Chemother Pharmacol. 1986;16(1):28–34. doi: 10.1007/BF00255282. [DOI] [PubMed] [Google Scholar]
  22. Rahman A., Husain S. R., Siddiqui J., Verma M., Agresti M., Center M., Safa A. R., Glazer R. I. Liposome-mediated modulation of multidrug resistance in human HL-60 leukemia cells. J Natl Cancer Inst. 1992 Dec 16;84(24):1909–1915. doi: 10.1093/jnci/84.24.1909. [DOI] [PubMed] [Google Scholar]
  23. Rahman A., Kessler A., More N., Sikic B., Rowden G., Woolley P., Schein P. S. Liposomal protection of adriamycin-induced cardiotoxicity in mice. Cancer Res. 1980 May;40(5):1532–1537. [PubMed] [Google Scholar]
  24. Rahman A., Treat J., Roh J. K., Potkul L. A., Alvord W. G., Forst D., Woolley P. V. A phase I clinical trial and pharmacokinetic evaluation of liposome-encapsulated doxorubicin. J Clin Oncol. 1990 Jun;8(6):1093–1100. doi: 10.1200/JCO.1990.8.6.1093. [DOI] [PubMed] [Google Scholar]
  25. Rahman A., White G., More N., Schein P. S. Pharmacological, toxicological, and therapeutic evaluation in mice of doxorubicin entrapped in cardiolipin liposomes. Cancer Res. 1985 Feb;45(2):796–803. [PubMed] [Google Scholar]
  26. Tan C., Etcubanas E., Wollner N., Rosen G., Gilladoga A., Showel J., Murphy M. L., Krakoff I. H. Adriamycin--an antitumor antibiotic in the treatment of neoplastic diseases. Cancer. 1973 Jul;32(1):9–17. doi: 10.1002/1097-0142(197307)32:1<9::aid-cncr2820320102>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  27. Thierry A. R., Jorgensen T. J., Forst D., Belli J. A., Dritschilo A., Rahman A. Modulation of multidrug resistance in Chinese hamster cells by liposome-encapsulated doxorubicin. Cancer Commun. 1989;1(5):311–316. doi: 10.3727/095535489820874931. [DOI] [PubMed] [Google Scholar]
  28. Thierry A. R., Rahman A., Dritschilo A. A new procedure for the preparation of liposomal doxorubicin: biological activity in multidrug-resistant tumor cells. Cancer Chemother Pharmacol. 1994;35(1):84–88. doi: 10.1007/BF00686289. [DOI] [PubMed] [Google Scholar]
  29. Thierry A. R., Vigé D., Coughlin S. S., Belli J. A., Dritschilo A., Rahman A. Modulation of doxorubicin resistance in multidrug-resistant cells by liposomes. FASEB J. 1993 Apr 1;7(6):572–579. doi: 10.1096/fasebj.7.6.8097173. [DOI] [PubMed] [Google Scholar]
  30. Treat J., Greenspan A., Forst D., Sanchez J. A., Ferrans V. J., Potkul L. A., Woolley P. V., Rahman A. Antitumor activity of liposome-encapsulated doxorubicin in advanced breast cancer: phase II study. J Natl Cancer Inst. 1990 Nov 7;82(21):1706–1710. doi: 10.1093/jnci/82.21.1706. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES