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Evaluation of incorporation characteristics of mitoxantrone into unilamellar liposomes and analysis of their pharmacokinetic properties, acute toxicity, and antitumor efficacy


Mitoxantrone (MTO) was incorporated into small unilamellar liposomes by formation of a complex between the anticancer drug and negatively charged lipids. The complex was formed at a 2:1 molar ratio between the lipids and MTO, with phosphatidic acid (PA) being the strongest complex-forming lipid. Weaker complexes and lower incorporation rates of MTO resulted when liposomes containing dicetylphosphate, phosphatidyl inositol, phosphatidyl serine, phosphatidyl glycerol, oleic acid, and tridecylphosphate were used. Thus, all further experiments were performed with PA-MTO liposomes that contained 0.1–3 mg MTO/ml and had mean vesicle sizes of 40–150 nm, depending on the drug concentration and the method of liposome preparation. In vitro incubations of free and liposomal MTO with human plasma showed that the drug is slowly transferred from the liposome membranes to the plasma proteins. For liposomal MTO a transfer rate of 48% was determined, whereas 75.8% of free MTO was bound to the plasma proteins. The organ distribution of the two preparations in mice showed that higher and longer-lasting concentrations of liposomal MTO were found in the liver and spleen. The terminal elimination halflives in the liver were 77 h for liposomal MTO and 14.4 h for free MTO. In the blood, slightly higher concentrations were detected for liposomal MTO, which also had slower biphasic elimination kinetics as compared with the free drug. Drug distribution in the heart was not significantly different from that in the kidneys. The LD25 of PA-MTO liposomes in mice was 19.6 mg/kg and that of free MTO was 7.7 mg/kg. The antitumor effects of PA-MTO liposomes were evaluated in murine L 1210 leukemia, in various xenografted human tumors, and in methylnitrosourea-induced rat mammary carcinoma. Generally, the liposomal application form was more effective and less toxic than the free drug. The cytostatic effects were dependent on the tumor model, the application schedule, and the drug concentration. At doses that were toxic when free MTO was used, the liposomal preparation produced strong antitumor effects in some cases. In summary, the incorporation of MTO into liposomes changes the drug's plasmabinding properties, alters its organ distribution, reduces its acute toxicity, and increases its cytostatic efficiency in various tumor models. The liposomal PA-MTO complex represents a new application form of MTO that has advantageous properties.

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  1. 1.

    Allen TM, Hansen C, Rutledge J (1989) Liposomes with prolonged circulation times: factors affecting uptake by reticuloendothelial and other tissues. Biochim Biophys Acta 981: 27–35

  2. 2.

    Balazsovits JAE, Mayer LD, Bally MB, Cullis PR, McDonell M, Ginsberg RS, Falk RE (1989) Analysis of the effect of liposome encapsulation on the vesicant properties, acute and cardiac toxicities, and antitumor efficacy of doxorubicin. Cancer Chemother Pharmacol 23: 81–86

  3. 3.

    Bartkowiak J, Kapuscinski J, Melamed MR, Darzynkiewicz Z (1989) Selective displacement of nuclear proteins by antitumor drugs affinity for nucleic acids. Proc Natl Acad Sci USA 86: 5151–5154

  4. 4.

    Berger MR, Floride J, Schmähl D, Schreiber J, Eisenbrand G (1986) Estrogen-linked 2-chloroethylnitrosoureas: anticancer efficacy in MNU-induced rat mammary carcinoma, uterine activity in mice and receptor interactions. Eur J Cancer Clin Oncol 22: 1179–1191

  5. 5.

    Brito-Babapulle F, Catovsky D, Newland AC, Goldman JM, Galton DAG (1987) Treatment of acute myeloid leukemia with intermediate-dose cytosine arabinoside and mitoxantrone. Semin Oncol 14 [Suppl 1]: 51–52

  6. 6.

    Burke TG, Tritton TR (1985) Structural basis of anthracyline selectivity for unilamellar phosphatidylcholine vesicles: an equilibrium binding study. Biochemistry 24: 1768–1776

  7. 7.

    Burke TG, Sartorelli AC, Tritton TR (1988) Selectivity of the anthracyclines for negatively charged model membranes: role of the amino group. Cancer Chemother Pharmacol 21: 274–280

  8. 8.

    Burns CP, Haugstad BN, Mossman CJ, North JA, Ingraham LM (1988) Membrane lipid alteration: effect on cellular uptake of mitoxantrone. Lipids 23: 393–397

  9. 9.

    Coltman CA, Coltman TM, Balcerzak SP, Morrison FS, Von Hoff DD (1984) Mitoxantrone in refractory non-Hodgkin's lymphoma. A Southwest Oncology Group study. Semin Oncol 11 [Suppl 1]: 50–53

  10. 10.

    Conrad MJ, Singer SJ (1981) The solubility of amphipathic molecules in biological membranes and lipid bilayers and its implications for membrane structure. Biochemistry 20: 808–818

  11. 11.

    Delgado G, Potkul RK, Treat JA, Lewandowski GS, Barter JF, Forst D, Rahman A (1989) A phase I/II study of intraperitoneally administered doxorubicin entrapped in cardiolipin liposomes in patients with ovarian cancer. Am J Obstet Gynecol 160: 812–819

  12. 12.

    Drewinko B, Yang L-Y, Barlogie B, Trujillo JM (1983) Comparative cytotoxicity of bisantrene, mitoxantrone, ametantrone, dihydroxyan-thracenedione, dihydroxy-anthracenedione diacetate, and doxorubicin on human cells in vitro. Cancer Res 43: 2648–2653

  13. 13.

    Durr FE (1984) Biologic and biochemical effects of mitoxantrone. Semin Oncol 11 [Suppl 1]: 3–10

  14. 14.

    Fiebig HH (1988) Comparison of tumor response in nude mice and in patients. In: Winograd B, Peckham MJ, Pineo HM (eds) Human tumor xenografts in anticancer drug development. Springer, Berlin Heidelberg New York London Paris Tokyo, pp 25–30

  15. 15.

    Fiebig HH, Löhr GW (1984) Wachstum menschlicher Malignome in der thymusaplastischen Nacktmaus. Med Welt 35: 52–58, 81–86

  16. 16.

    Ford JM, Panasci L, Leclerc Y, Margolese R (1987) Phase II trial of a combination of doxorubicin and mitoxantrone in metastatic breast cancer. Cancer Treat Rep 71: 921–925

  17. 17.

    Fortmeyer HP, Bastert G (1981) Breeding and maintenance of nu/nu mice and rnu/rnu rats. In: Bastert G, Fortmeyer HP, Schmidt-Mathiesen H (eds) Thymus aplastic nude mice and rats in clinical oncology. G. Fischer, Stuttgart New York, pp 25–38

  18. 18.

    Gabizon A, Papahadjopoulos D (1989) Liposome formulations with prolonged circulation time in blood and enhanced uptake by tumors. Proc Natl Acad Sci USA 85: 6949–6953

  19. 19.

    Gabizon A, Sulkes A, Peretz T, Druckmann S, Goren D, Amselem S, Barenholz Y (1989) Liposome-associated doxorubicin: preclinical pharmacology and exploratory clinical phase I/II. In: Lopez-Berenstein G, Fidler IJ (eds) Liposomes in the therapy of infectious diseases and cancer. Alan R. Liss, New York, pp 391–407

  20. 20.

    Goormaghtigh E, Chatelain P, Caspers J, Ruysschaert JM (1980) Evidence of a specific complex between Adriamycon and negatively-charged phospholipids. Biochim Biophys Acta 597: 1–14

  21. 21.

    Grant Prentice H, Robbins G, Ma DDF, Ho AD (1984) Mitoxantrone in relapsed and refractory acute leukemia. Semin Oncol 11 [Suppl 1]: 32–35

  22. 22.

    Heath TD, Lopez NG, Piper JR, Montgomery JA, Stern WH, Papahadjopoulos D (1986) Liposome-mediated delivery of pteridine antifolates to cells in vitro: potency of methotrexate, and its alpha and gamma substituents. Biochim Biophys Acta 862: 72–86

  23. 23.

    Hiddemann W, Kreutzmann H, Straif K, Ludwig W-D, Mertelsmann R, Planker M, Donhuijsen-Ant R, Lengfelder E, Arlin Z, Büchner T (1984) High-dose cytosine arabinoside in combination with mitoxantrone for the treatment of refractory acute myeloid and lymphoblastic leukemia. Semin Oncol 14 [Suppl 1]: 73–77

  24. 24.

    Hoesel QCGM van, Steerenberg PA, Crommelin DJA, Dijk A van, Oort W van, Klein S, Douze JMC, Wildt DJ de, Hillen FC (1984) Reduced cardiotoxicity and nephrotoxicity with preservation of antitumor activity of doxorubicin entrapped in stable liposomes in the LOU/M Wsl rat. Cancer Res 44: 3698–3705

  25. 25.

    Johnson RK, Zee-Cheng RK-Y, Lee WW, Acton EM, Henry DW, Cheng CC (1979) Experimental antitumor activity of anthraquinones. Cancer Treat Rep 63: 425–439

  26. 26.

    Kapuscinski J, Darzynkiewicz Z, Traganos F, Melamed MR (1981) Interactions of a new antitumor agent, 1,4-dihydroxy-5,8-bis[[2-[(2-hydroxyethyl)amino]-ethyl]amino]-9,10-anthracenedione, with nucleic acids. Biochem Pharmacol 30: 231–240

  27. 27.

    Khokhar AR, Al-Baker S, Krakoff IH, Perez-Soler R (1989) Toxicity and antitumor activity ofcis-bis-carboxylato(trans-R,R-1,2-diaminocyclohexane) platinum(II) complexes entrapped in liposomes. Cancer Chemother Pharmacol 23: 219–224

  28. 28.

    Landys K, Borgstrom S, Andersson T, Noppa H (1985) Mitoxantrone as a first-line treatment of advanced breast cancer. Invest New Drugs 3: 133–137

  29. 29.

    Lu K, Savaraj N, Loo TL (1984) Pharmacological disposition of 1,4-dihydroxy-5,8-bis[[2-[(2-hydroxyethyl)amino]ethyl]amino]-9,10-anthracenedione dihydrochloride in the dog. Cancer Chemother Pharmacol 13: 63–66

  30. 30.

    Mayhew EG, Goldrosen MH, Vaage J, Rustum YM (1987) Effects of liposome-entrapped doxorubicin on liver metastases of mouse colon carcinomas 26 and 28. J Natl Cancer Inst 78: 707–713

  31. 31.

    Paciucci PA, Cuttner J, Holland JF (1984) Mitoxantrone as a single agent and in combination chemotherapy in patients with refractory acute leukemia. Semin Oncol 11 [Suppl 1]: 36–40

  32. 32.

    Patel KR, Baldeschwieler JD (1984) Treatment of intravenously implanted Lewis lung carcinoma with liposome-encapsulated cytosine arabinoside and non-specific immunotherapy. Int J Cancer 34: 415–420

  33. 33.

    Perez-Soler R, Khokhar AR, Lopez-Berenstein G (1987) Treatment and prophylaxis of experimental liver metastases of M5076 reticulosarcoma withcis-bis-neodecanato-trans-R,R-1,2-diaminocyclohexane platinum(II) encapsulated in multilamellar vesicles. Cancer Res 47: 6462–6466

  34. 34.

    Pestalozzi B, Schwendener RA, Sauter Chr (1990) Liposomal mitoxantrone (L-Mto) is effective in metastatic breast cancer (abstract). Proc Am Soc Clin Oncol 9: 26

  35. 35.

    Rahman A, Fumagalli A, Barbieri B, Schein P, Casazza AM (1986) Antitumor and toxicity evaluation of free doxirubicin and doxorubicin entrapped in cardiolipin liposomes. Cancer Chemother Pharmacol 16: 22–27

  36. 36.

    Richard B, Fabre G, Fabre I, Cano JP (1989) Excretion and metabolism of mitoxantrone in rabbits. Cancer Res 49: 833–837

  37. 37.

    Rubas W, Supersaxo A, Weder HG, Hartmann HR, Hengartner H, Schott H, Schwendener R (1986) Treatment of murine L1210 lymphoid leukemia and melanoma B 16 with lipophilic cytosine arabinoside prodrugs incorporated into unilamellar liposomes. Int J Cancer 37: 149–154

  38. 38.

    Schwendener RA (1986) The preparation of large volumes of homogeneous, sterile liposomes containing various lipophilic cytostatic drugs by the use of a capillary dialyzer. Cancer Drug Deliv 3: 123–129

  39. 39.

    Schwendener RA (1988) Incorporation of chlorpromazine into bilayer liposomes for protection against microsomal metabolism and liver absorption. Eur J Drug Metab Pharmacokinet 13: 135–141

  40. 40.

    Schwendener RA, Weder HG (1978) The binding of chlorpromazine to bilayer liposomes. Biochem Pharmacol 27: 2721–2727

  41. 41.

    Schwendener RA, Schott H, Hartmann HR, Supersaxo A, Rubas W, Hengartner H (1987) Liposomen al Träger von lipophilen n Cytosinarabinosid- und Fluorodeoxyuridin-Derivaten. Onkologie 10: 232–239

  42. 42.

    Schwendener RA, Pestalozzi B, Berger S, Schott H, Hengartner H, Sauter C (1989) Treatment of acute myelogenous leukemia with liposomes containingN 4-oleyl-cytosine arabinoside. In: Lopez-Berenstein G, Fidler IJ (eds) Liposomes in the therapy of infectious diseases and cancer. Alan R. Liss, New York, pp 95–103

  43. 43.

    Shenkenberg TD, Von Hoff DD (1986) Mitoxantrone: a new anticancer drug with significant clinical activity. Ann Intern Med 105: 67–81

  44. 44.

    Smith IE (1983) Mitoxantrone (Novantrone): a review of experimental and early clinical studies. Cancer Treat Rev 10: 103–115

  45. 45.

    Steerenberg PA, Storm G, Groot G de, Claessen A, Bergers JJ, Franken MAM, Hoesel QGCM van, Wubs KL, Jong WH de (1988) Liposomes as drug carrier system forcis-diamminedichloroplatinum (II): II. Antitumor activity in vivo, induction of drug resistance, nephrotoxicity and Pt distribution. Cancer Chemother Pharmacol 21: 299–307

  46. 46.

    Stuart-Harris RC, Bozek T, Pavlidis NA, Smith IE (1984) Mitoxantrone: an active new agent in the treatment of advanced breast cancer. Cancer Chemother Pharmacol 12: 1–4

  47. 47.

    Supersaxo A, Rubas W, Hartmann HR, Schott H, Hengartner H, Schwendener RA (1988) The antitumor effect of lipophilic derivatives of 5-fluoro-2′-deoxyridine incorporated into liposomes. J Microencaps 5: 1–11

  48. 48.

    Vile GF, Winterbourn CC (1989) Microsomal lipid peroxidation induced by Adriamycin, epirubicin, daunorubicin and mitoxantrone: a comparative study. Cancer Chemother Pharmacol 24: 105–108

  49. 49.

    Wallace RE, Murdock KC, Angier RB, Durr FE (1979) Activity of a novel anthracenedione, 1,4-dihydroxy-5,8-bis{{{2-[(2-hydroxyethyl)amino]ethyl}amino}}-9,10-anthracenedione dihydrochloride, against experimental tumors in mice. Cancer Res 39: 1570–1574

  50. 50.

    Walters RS, Kantarjian HM, Keating MJ, Plunkett WK, Estey EH, Andersson B, Beran M, McCredie KB, Freireich EJ (1988) Mitoxantrone and high-dose cytosine arabinoside in refractory acute myelogenous leukemia. Cancer 62: 677–682

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Correspondence to Reto A. Schwendener.

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This research was supported in part by the Krebsliga of the Kanton Zürich and by Lederle Arzneimittel, Wolfratshausen, Federal Republic of Germany

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Schwendener, R.A., Fiebig, H.H., Berger, M.R. et al. Evaluation of incorporation characteristics of mitoxantrone into unilamellar liposomes and analysis of their pharmacokinetic properties, acute toxicity, and antitumor efficacy. Cancer Chemother. Pharmacol. 27, 429–439 (1991). https://doi.org/10.1007/BF00685156

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  • Mitoxantrone
  • Free Drug
  • Phosphatidic Acid
  • Phosphatidic Acid
  • Phosphatidyl Inositol