Skip to main content

Advertisement

SpringerLink
Log in

The preparation and characterization of anti-VEGFR2 conjugated, paclitaxel-loaded PLLA or PLGA microspheres for the systemic targeting of human prostate tumors

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

The objective of this study was to manufacture paclitaxel (PTX) loaded polymeric microspheres, that were surface conjugated with antibodies to vascular endothelial growth factor receptor 2 (anti-VEGFR2), for systemic targeting to angiogenic sites in prostate tumors.

Methods

Microspheres were manufactured in the 1–3 μm size range from poly (l-lactic acid) (PLLA) or poly (lactide-co-glycolide) (PLGA) by a modified solvent evaporation method using Polytron homogenization followed by high speed dispersion in poly vinyl alcohol. Antibodies were conjugated to the surface of these microspheres using cyanogen bromide activation of the polymer surface. Cell Binding was determined using human umbilical vein endothelial cells (HUVECs) in vitro. Efficacy determinations were made using human prostate tumors (PC-3) grown subcutaneously in mice.

Results

Antibodies were effectively bound to the surface of PLLA and PLGA micropsheres. Anti-VEGFR2 conjugated PLLA microspheres bound strongly to HUVEC’s. Pilot efficacy studies in mice showed variability but demonstrated a significant inhibition of tumor growth following the systemic administration of a single dose of PTX-loaded anti-VEGFR2 conjugated PLLA microspheres as compared to non-antibody-conjugated PTX-loaded microspheres.

Conclusion

Anti-VEGFR2 conjugated PLLA microspheres containing PTX may offer an effective way of administering a controlled release formulation of the drug to target prostate tumors.

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
Fig. 7

Similar content being viewed by others

References

  1. Arshady R, Monshipouri M (eds) (1999) Targeted delivery of microparticulate carriers. Microspheres, microcapsules, and liposomes: medical and biotechnology applications, vol 2. Imperial College, University of London, Citus Books, London

  2. Balbay MD, Pettaway CA, Kuniyasu H, Inouse K, Ramirez E, Li E, Fidler IJ, Dinney CP (1999) Highly metastatic human prostate cancer growing within the prostate of athymic mice overexpresses vascular endothelial growth factor. Clin Cancer Res 5(4):783–789

    PubMed  CAS  Google Scholar 

  3. Berkland C, Kyekyoon K, Pack DW (2001) Fabrication of PLG microspheres with precisely controlled and monodisperse size distributions. J Control Release 73:59–74

    Article  PubMed  CAS  Google Scholar 

  4. Bitz C, Doelker E (1996) Influence of the preparation method on residual solvents in biodegradable microspheres. Int J Pharm 131:171–181

    Article  CAS  Google Scholar 

  5. Brekken RA, Thorpe PE (2001) VEGF–VEGF receptor complexes as markers of tumor vascular endothelium. J Control Release 74:173–181

    Article  PubMed  CAS  Google Scholar 

  6. Dhanaraj SA, Santhi K, Pandit V, Raghu R, Vijayan P, Vasudevan M, Suresh B (2005) Study on the active delivery of methotrexate microspheres with mouse monoclonal IgG in tumour-induced mice. Indian J Pharm Sci 67:540–543

    CAS  Google Scholar 

  7. Dordunoo SK, Jackson JK, Arsenault LA, Oktaba AMC, Hunter WC, Burt HM (1995) Taxol encapsulation in poly(caprolactone) microspheres. Cancer Chemother Pharmacol 36:279–282

    PubMed  CAS  Google Scholar 

  8. Ferrara N, Hillan KJ, Novotny W (2005) Bevacizumab (Avastin), a humanized anti-VEGF monoclonal antibody for cancer therapy. Biochem Biophys Res Commun 333:328–335

    Article  PubMed  CAS  Google Scholar 

  9. Fonseca C, Simoes S, Gaspar R (2002) Paclitaxel loaded PLGA nanoparticles: preparation, physiochemical characterization and in vitro anti-tumoral activity. J Control Release 83:273–286

    Article  PubMed  CAS  Google Scholar 

  10. Freitas S, Merkle HP, Gander B (2005) Microencapsulation by solvent extraction/evaporation: reviewing the state of the art of microsphere preparation process technology. J Control Release 102:313–332

    Article  PubMed  CAS  Google Scholar 

  11. Gille H, Kowalski J, Li B, LeCouter J, Moffat B, Zioncheck TF, Pelletier N, Ferrara N (2001) Analysis of biological effects and signaling properties of Flt-1 (VEGFR-1) and KDR (VEGFR-2). A reassessment using novel receptor-specific vascular endothelial growth factor mutants. J Biol Chem 276(5):3222–3230

    Article  PubMed  CAS  Google Scholar 

  12. Gupte A, Ciftci K (2004) Formulation and characterization of Paclitaxel, 5-FU and Paclitaxel + 5-FU microspheres. Int J Pharm 276:93–106

    Article  PubMed  CAS  Google Scholar 

  13. Hermanson GT, Mallia K, Smith PK (eds) (1992) Immobilized affinity ligand techniques. Academic, New York

    Google Scholar 

  14. Hou JM, Liu JY, Yang L, Zhao X, Tian L, Ding ZY, Wen YJ, Niu T, Xiao F, Lou YY, Tan GH, Deng HX, Li J, Yang JL, Mao YQ, Kan B, Wu Y, Li Q, Wei YQ (2005) Combination of low dose gemcitabine and recombinant quail vascular endothelial growth factor receptor-2 as a vaccine induces synergistic antitumor activities. Oncology 69(1):81–87

    Article  PubMed  CAS  Google Scholar 

  15. Huang YQ, Li JJ, Hu L, Lee M, Karpatkin S (2001) Thrombin induces increased expression and secretion of VEGF from human FS4 fibroblasts, DU145 prostate cells and CHRF megakaryocytes. Thromb Haemost 86(4):1094–1098

    PubMed  CAS  Google Scholar 

  16. Jackson JK, Gleave JE, Yago V, Beraldi E, Hunter WL, Burt HM (2000) The suppression of human prostate tumor growth in mice by the intra-tumoral injection of a slow release polymeric paste formulation of paclitaxel. Cancer Res 60:4146–4151

    PubMed  CAS  Google Scholar 

  17. Jackson JK, Springate CMK, Hunter WL, Burt HM (2000) Neutrophil activation by plasma opsonized polymeric microspheres: inhibitory effect of Pluronic F127. Biomaterials 21:1483–1491

    Article  PubMed  CAS  Google Scholar 

  18. Keegan ME, Royce SM, Fahmy T, Saltzman WM (2006) In vitro evaluation of biodegradable microspheres with surface-bound ligands. J Control Release 110:574–580

    Article  PubMed  CAS  Google Scholar 

  19. Klement G, Baruchel S, Rak J, Man S, Clark K, Hicklin DJ, Bohlen P, Kerbel RS (2000) Continuous low dose therapy with vinblastine and VEGF receptor-2 antibody induces sustained tumor regression without overt toxicity. J Clin Invest 105(8):R15–R24

    Article  PubMed  CAS  Google Scholar 

  20. Kollerman J, Helpap B (2001) Expression of vascular endothelial growth factor (VEGF) and VEGF receptor Flk-1 in benign, premalignant, and malignant prostate tissue. Am J Clin Pathol 116(1):115–121

    Article  Google Scholar 

  21. Liggins RT, Burt HM (2001) Paclitaxel loaded poly(l-lactic acid) microspheres: properties of microspheres made with low molecular weight polymers. Int J Pharm 222:19–33

    Article  PubMed  CAS  Google Scholar 

  22. Liggins RT, D’Amours S, Demetrick JS, Machan LS, Burt HM (2000) Paclitaxel loaded poly(l-lactic acid) microspheres for the prevention of intraperitoneal carcinomatosis after a surgical repair and tumor cell spill. Biomaterials 21:1959–1969

    Article  PubMed  CAS  Google Scholar 

  23. Liggins RT, Cruz T, Min W, Liang L, Hunter WL, Burt HM (2004) Intra-articular treatment of arthritis with microsphere formulations of paclitaxel: biocompatibility and efficacy determinations in rabbits. Inflamm Res 53:363–372

    Article  PubMed  CAS  Google Scholar 

  24. Ma L, Francia G, Viloria-Petit A, Hicklin DJ, du Manoir J, Rak J, Kerbel RS (2005) In vitro procoagulant activity induced in endothelial cells by chemotherapy and antiangiogenic drug combinations: modulation by lower-dose chemotherapy. Cancer Res 65(12):5365–5373

    Article  PubMed  CAS  Google Scholar 

  25. Maruyama K (2000) In vivo targeting of liposomes. Biol Pharm Bull 23(7):791–799

    PubMed  CAS  Google Scholar 

  26. Molema G, Meijer DK, de Leij LF (1998) Tumor vasculature targeted therapies: getting the players organized. Biochem Pharmacol 55(12):1939–1945

    Article  PubMed  CAS  Google Scholar 

  27. Muramatsu N, Kondo T (1995) An approach to prepare microparticles of uniform size. J Microencapsul 12(2):129–136

    Article  PubMed  CAS  Google Scholar 

  28. Onetto N, Canetta R, Winograd R (1993) Overview of taxol safety. J Natl Cancer Inst Monogr 15:131–139

    PubMed  Google Scholar 

  29. Ramaswamy M, Zhang X, Burt HM, Wasan (1997) The human plasma distribution of free paclitaxel and paclitaxel associated with diblock copolymers. J Pharm Sci 86:460–464

    Article  PubMed  CAS  Google Scholar 

  30. Scholes PD, Coombes L, Illum L, Davis SS, Watts JF, Ustariz C, Vert M, Davies MC (1999) Detection and determination of surface levels of poloxamer and PVA surfactant on biodegradable microspheres using SSIMS and XPS. J Control Release 59:261–278

    Article  PubMed  CAS  Google Scholar 

  31. Sharma A, Straubinger RM (1994) Novel taxol formulations: preparation and characterization of taxol-containing liposomes. Pharm Res 11:889–896

    Article  PubMed  CAS  Google Scholar 

  32. Springate CM, Jackson JK, Gleave ME, Burt HM (2005) Efficacy of an intra-tumoral controlled release formulation of clusterin antisense oligonucleotide complexed with chitosan containing paclitaxel or docetaxel in prostate cancer xenograft models. Cancer Chemother Pharmacol 56(3):239–247

    Article  PubMed  CAS  Google Scholar 

  33. Torchilin VP, Lukyanov AN, Gao Z, Papahadjopoulos-Sternberg B (2003) Immunomicelles: targeted pharmaceutical carriers for poorly soluble drugs. Proc Natl Acad Sci USA 100(10):6039–6044

    Article  PubMed  CAS  Google Scholar 

  34. Witte L, Hicklin DJ, Zhu Z, Pytowski B, Kotanides H, Rockwell P, Bohlen P (1998) Monoclonal antibodies targeting the VEGF receptor-2 (Flk1/KDR) as an anti-angiogenic therapeutic strategy. Cancer Metastasis Rev 17:155–161

    Article  PubMed  CAS  Google Scholar 

  35. Zhifang Z, Mingxing Z, Shenghao W, Fang L, Wenzhao S (1993) Preparation and evaluation in vitro and vivo of copoly (lactic/glycolic) acid microspheres containing norethisterone. Biomater Artif Cells Immobilization Biotechnol 21(1):71–84

    Google Scholar 

Download references

Acknowledgments

The authors would like to thank the Canadian Prostate Cancer Research Initiative Ideas Grant program for providing funding for this project. The authors would also like to acknowledge Virginia Yago in Dr Gleave’s group for performing the animal experiments.

Author information

Authors and Affiliations

  1. Division of Pharmaceutics and Biopharmaceutics, Faculty of Pharmaceutical Sciences, The University of British Columbia, 2146 East Mall, Vancouver, BC, Canada, V6T 1Z3

    Jianjun Lu, John K. Jackson & Helen M. Burt

  2. Division of Urology, The Prostate Centre, Vancouver General Hospital, The University of British Columbia, Vancouver, BC, Canada, V6H 3Z3

    Martin E. Gleave

Authors
  1. Jianjun Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John K. Jackson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Martin E. Gleave
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Helen M. Burt
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Helen M. Burt.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lu, J., Jackson, J.K., Gleave, M.E. et al. The preparation and characterization of anti-VEGFR2 conjugated, paclitaxel-loaded PLLA or PLGA microspheres for the systemic targeting of human prostate tumors. Cancer Chemother Pharmacol 61, 997–1005 (2008). https://doi.org/10.1007/s00280-007-0557-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-007-0557-x

Keywords

Search

Navigation