Cancer Chemotherapy and Pharmacology

, Volume 55, Issue 6, pp 565–576 | Cite as

Improvement of paclitaxel therapeutic index by derivatization and association to a cholesterol-rich microemulsion: in vitro and in vivo studies

  • Debora G. Rodrigues
  • Durvanei A. Maria
  • Denise C. Fernandes
  • Claudete J. Valduga
  • Ricardo D. Couto
  • Olga C. M. Ibañez
  • Raul C. Maranhão
Original Article

Abstract

A cholesterol-rich microemulsion or nanoparticle termed LDE concentrates in cancer tissues after injection into the bloodstream. Here the cytotoxicity, pharmacokinetics, toxicity to animals and therapeutic action of a paclitaxel lipophilic derivative associated to LDE is compared with those of the commercial paclitaxel. Results show that LDE-paclitaxel oleate is stable. The cytostatic activity of the drug in the complex is diminished compared with the commercial paclitaxel due to the cytotoxicity of the vehicle Cremophor EL used in the commercial formulation. Competition experiments in neoplastic cultured cells show that paclitaxel oleate and LDE are internalized together by the LDL receptor pathway. LDE-paclitaxel oleate arrests the G2/M phase of cell cycle, similarly to commercial paclitaxel. Tolerability to mice is remarkable, such that the lethal dose (LD50) was ninefold greater than that of the commercial formulation (LD50 = 326 μM and 37 μM, respectively). LDE concentrates paclitaxel oleate in the tumor roughly fourfold relative to the normal adjacent tissues. At equimolar doses, the association of paclitaxel oleate with LDE results in remarkable changes in the drug pharmacokinetic parameters when compared to commercial paclitaxel (t1/2=218 min and 184 min, AUC=1,334 μg h/ml and 707 μg h/ml and CL=0.125 ml/min and 0.236 ml/min, respectively). Finally, the therapeutic efficacy of the complex is pronouncedly greater than that of the commercial paclitaxel, as indicated by the reduction in tumor growth, increase in survival rates and % cure of treated mice. In conclusion, LDE-paclitaxel oleate is a stable complex and compared with paclitaxel toxicity is considerably reduced and activity is enhanced, which may lead to improved therapeutic index in clinical use.

Keywords

Nanoparticles Paclitaxel Emulsions Cholesterol Low-density lipoprotein receptors Cancer treatment Drug targeting 

Notes

Acknowledgments

This study was supported by Fundação do Amparo à Pesquisa do Estado de São Paulo (FAPESP), São Paulo, Brazil (Grant 99/01229-2). Dr Maranhão has a Research Award from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brasilia, Brazil.

References

  1. 1.
    Ades A, Carvalho JP, Graziani SR, Amancio RF, Souen JS, Pinotti JA, Maranhão RC (2001) Uptake of a cholesterol-rich emulsion by neoplastic ovarian tissues. Gynecol Oncol 82:84–87CrossRefGoogle Scholar
  2. 2.
    Constantinides PP, Lambert KJ, Tustian AK, Shneider B, Lalji S, Ma W, Wentzel B, Kesller D, Worah D, Quay SC (2000) Formulation development and antitumor activity of a filter-sterilizable emulsion of paclitaxel. Pharm Res 17(2):175–182CrossRefGoogle Scholar
  3. 3.
    Csóka K, Dhar S, Fridborg H, Larsson R, Nygren P (1997) Differential activity of cremophor EL and paclitaxel in patient tumor cells and human carcinoma cell lines in vitro. Cancer 79:1225–1233CrossRefPubMedGoogle Scholar
  4. 4.
    Entin I, Plotnikov A, Korenstein R, Keisari Y (2003) Tumor growth retardation, cure, and induction of antitumor immunity in B16 Melanoma-bearing mice by low eletric field-enhanced chemotherapy. Clin Cancer Res 9:3190–3197Google Scholar
  5. 5.
    Gal D, Ohashi M, MacDonald PC, Buchsbaum HJ, Simpson ER (1981) Low-density lipoprotein as a potential vehicle for chemotherapeutic agents and radionucleotides in the management of gynecologic neoplasms. Am J Obstret Gynecol 139:877–885Google Scholar
  6. 6.
    Ginsberg H, Gilbert HS, Gibson JC, Ngoc-Anh L, Virgil BW (1986) Increased low-density-lipoprotein catabolism in myeloproliferative disorders. Ann Intern Med 96: 311–316Google Scholar
  7. 7.
    Ginsburg GS, Small DM, Atkinson D (1982) Microemulsions of phospholipids and cholesterol esters. Protein-free models of low density lipoprotein. J Biol Chem 57:8216–8277Google Scholar
  8. 8.
    Goble S, Bear HD (2003) Emerging role of taxanes in adjuvant and neoadjuvant therapy for breast cancer: the potential and the questions. Surg Clin North Am 83(4):943–971Google Scholar
  9. 9.
    Graziani SR, Igreja FAF, Hegg R, Meneghetti C, Brandizzi LI, Barboza R, Amâncio RF, Pinotti JA, Maranhão RC (2002) Uptake of a cholesterol-rich emulsion by breast cancer. Gynecol Oncol 85:493–497CrossRefGoogle Scholar
  10. 10.
    Gueddari N, Favre G, Hachem H, Marek E, Le Gaillard F, Soula G (1993) Evidence for up-regulated low density lipoprotein receptor in human lung adenocarcinoma cell line A549. Biochemie 75:811–819CrossRefGoogle Scholar
  11. 11.
    Henriksson P, Ericsson S, Stege R, Ericsson M, Rudling M, Berglund L, Angelin B (1989) Hypocholesterolaemia and increased elimination of low-density lipoproteins in metastatic cancer of the prostate. Lancet 2:1178–1180CrossRefGoogle Scholar
  12. 12.
    Hungria VTM, Latrilha MC, Rodrigues DG, Bydlowski SP, Chiattone CS, Maranhão RC (2004) Metabolism of a cholesterol-rich microemulsion (LDE) in patients with multiple myeloma and a preliminary clinical study of LDE as a drug vehicle for the treatment of the disease. Cancer Chem Pharmacol 53:51–60CrossRefGoogle Scholar
  13. 13.
    Kalechman YK, Longo DL, Catane R, Shasi A, Albeck M, Serdini B (2000) Synergistic anti-tumoral effect of paclitaxel (Taxol)+AS101 in a murine model of B16 melanoma: association with ras-dependent signal transduction pathways. Int J Cancer 86:281–288CrossRefGoogle Scholar
  14. 14.
    Lundberg BB, Risovic V, Ramaswamy M, Wasan KM (2003) A lipophilic paclitaxel derivative incorporated in a lipid emulsion for parenteral administration. J Control Release 86:93–100CrossRefPubMedGoogle Scholar
  15. 15.
    Maranhão RC, Cesar TB, Pedroso MTB, Hirata MH, Mesquita CH (1993) Metabolic behavior in rats of a nonprotein microemulsion resembling LDL. Lipids 28:691–696Google Scholar
  16. 16.
    Maranhão RC, Garicochea B, Silva EL, Dorlhiac-Llacer P, Cadena SMS, Coelho IJC, Meneghetti JC, Pileggi FJC, Chamone DAF (1994) Plasma kinetics and biodistribution of a lipid emulsion resembling low-density lipoprotein in patients with acute leukemia. Cancer Res 54:4660–4666Google Scholar
  17. 17.
    Maranhão RC, Graziani SR, Yamaguchi N, Melo RF, Latrilha MC, Rodrigues DG, Couto RD, Schreier S, Buzaid AC (2002) Association of carmustine with a lipid emulsion: in vitro, in vivo and preliminary studies in cancer patients. Cancer Chem Pharmacol 49(6):487–498CrossRefGoogle Scholar
  18. 18.
    Matthews CME (1957) The theory of tracer experiments with 1331 I-labeled plasma proteins. Phys Med Biol 2:36–44CrossRefGoogle Scholar
  19. 19.
    Plowman J, Dykes DJ, Hollingshead M, Simpson H, Alley MC (1997) Human tumor xenograft models in NCI development. In: Teicher BA (ed) Anticancer drug development: preclinical screening, clinical trials and approval. Humana, Totowa, pp 101–125Google Scholar
  20. 20.
    Reinecke P, Corvin J, Gabbert HE, Gerharz CD (1996) Antiproliferative effects of paclitaxel (taxol) on human renal clear cell carcinomas in vitro. Eur J Cancer 33:1122–1129CrossRefGoogle Scholar
  21. 21.
    Rodrigues DG, Covolan CC, Coradi ST, Barboza R, Maranhão RC (2002) Use of a cholesterol-rich emulsion that binds to low-density lipoprotein receptors as a vehicle for paclitaxel. J Pharm Phamacol 56:765–772CrossRefGoogle Scholar
  22. 22.
    Sharma A, Mayhew E, Bolcsak L, Cavanaugh C, Harmon P, Janoff A, Bernacki RJ (1997) Activity of paclitaxel liposome formulations against human ovarian tumor xenografts. Int J Cancer 71:103–107CrossRefGoogle Scholar
  23. 23.
    Ueyama Y, Matsuzawa U, Yamashita S, Funahashi T, Sakai N, Nakamura T, Kubo M, Tarui S (1990) Hypocholesterolemic factor from gallbladder cancer cells. Lancet 336:707–709CrossRefGoogle Scholar
  24. 24.
    Valduga CJ, Fernandes DC, Lo Prete AC, Azevedo CHM, Rodrigues DG, Maranhão RC (2003) Use of a cholesterol-rich microemulsion that binds to low-density lipoprotein receptors as vehicle for etoposide. J Pharm Pharmacol 55:1615–1622CrossRefGoogle Scholar
  25. 25.
    Verluis AJ, Rensen PC, Rump ET, Van Berkel TJ, Bijsterbosch MK (1998) Low density lipoprotein receptor-mediated delivery of a lipophilic daunorubicin derivative to B16 tumours in mice using apolipoprotein E-enriched liposomes. Br J Cancer 78(12):1607–1614Google Scholar
  26. 26.
    Wang T, Wang H, Soong Y (2000) Paclitaxel-induced cell death. Cancer 88(11):2619–2628CrossRefGoogle Scholar
  27. 27.
    Weiss RB, Donehower RC, Wiernik PH, Ohnuma T, Gralla RJ, Trump DL, Baker JL, VanEcho DA, VonHoff DD, Leyland J (1990) Hypersensitivity reactions from taxol. J Clin Oncol 8:1263–1268PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Debora G. Rodrigues
    • 1
    • 2
  • Durvanei A. Maria
    • 3
  • Denise C. Fernandes
    • 1
  • Claudete J. Valduga
    • 1
  • Ricardo D. Couto
    • 1
    • 2
  • Olga C. M. Ibañez
    • 3
  • Raul C. Maranhão
    • 1
    • 2
  1. 1.Lipid Metabolism Laboratory, Heart Institute (InCor)University of São Paulo Medical School HospitalSao PauloBrazil
  2. 2.Faculty of Pharmaceutical SciencesUniversity of São PauloSao PauloBrazil
  3. 3.Laboratory of Immunogenetics of the Butantan InstituteSao PauloBrazil

Personalised recommendations