Pharmaceutical Research

, Volume 23, Issue 4, pp 782–789 | Cite as

Osmotic-Driven Release Kinetics of Bioactive Therapeutic Proteins from a Biodegradable Elastomer are Linear, Constant, Similar, and Adjustable

Research Paper

Purpose

The aim of the study is to determine whether a biodegradable elastomeric device that uses an osmotic pressure delivery mechanism can release different therapeutic proteins at a nearly constant rate in nanomolar concentrations with high bioactivity, given the same formulation conditions. Vascular endothelial growth factor (VEGF) and interleukin-2 (IL-2) were embedded in the device as sample therapeutic proteins, and their release and bioactivity were compared to that achieved previously with interferon-γ (IFN-γ).

Methods

A photo-cross-linkable biodegradable macromer consisting of acrylated star(ɛ-caprolactone-co-d,l-lactide) was prepared. VEGF, IL-2, and IFN-γ were co-lyophilized with serum albumin and trehalose at different ratios and were then embedded into the elastomer by photo-cross-linking the lyophilized particles in a macromer solution. The protein mass and the bioactivity in the release supernatant were measured by enzyme-linked immunosorbent and cell-based assays.

Results

VEGF, IL-2, and IFN-γ were released at the same, nearly constant rate of 25.4 ng/day for over 18 days. Using the optimum elastomer formulation, the release profiles of the proteins were essentially identical, and their rates were linear and constant. Cell-based bioactivity assays showed that 70 and 88% of the released VEGF and IL-2, respectively, were bioactive. The rate of protein release can be adjusted by changing the trehalose loading concentration in the elastomer matrix without altering the linear nature of the protein release kinetics. The elastomeric device degraded in PBS buffer within 85 days.

Conclusions

The elastomer formulation shows promising potential as a sustained protein drug delivery vehicle for local delivery applications.

Abbreviations

ASCP

acrylated star copolymer

IFN-γ

interferon-γ

IL-2

interleukin-2

SCP

star copolymer

VEGF

vascular endothelial growth factor

References

  1. 1.
    Sinha, V. R., Trehan, A. 2003Biodegradable microspheres for protein deliveryJ. Control. Release90261280CrossRefPubMedGoogle Scholar
  2. 2.
    Weert, M., Hennink, W. E., Jiskoot, W. 2000Protein instability in poly(lactic-co-glycolic acid) microparticlesPharm. Res.1711591167CrossRefPubMedGoogle Scholar
  3. 3.
    Lemaire, V., Belair, J., Hildgen, P. 2003Structural modeling of drug release from biodegradable porous matrices based on a combined diffusion/erosion processInt. J. Pharm.25895107CrossRefPubMedGoogle Scholar
  4. 4.
    Sah, H., Toddywala, R., Chien, Y. W. 1995Continuous release of proteins from biodegradable microcapsules and in-vivo evaluation of their potential as a vaccine adjuvantJ. Control. Release35137144CrossRefGoogle Scholar
  5. 5.
    Lee, W. K., Park, J. Y., Yang, E. H., Suh, H., Kim, S. H., Chung, D. S., Choi, K., Yang, C. W., Park, J. S. 2002Investigation of the factors influencing the release rates of cyclosporin A-loaded micro- and nanoparticles prepared by high-pressure homogenizerJ. Control. Release84115123CrossRefPubMedGoogle Scholar
  6. 6.
    Sendil, D., Wise, D. L., Hasirci, V. 2002Assessment of biodegradable controlled release rod systems for pain relief applicationsJ. Biomater. Sci. Polym. Ed.13115CrossRefPubMedGoogle Scholar
  7. 7.
    Hong, K. D., Ahn, Y. S., Go, J. T., Kim, M. S., Yuk, S. H., Shin, H. S., Rhee, J. M., Khang, G., Lee, H. B. 2005Preparation of double layered nanosphere using dextran and poly(l-lactide-co-glycolide)Polym-Korea29260265Google Scholar
  8. 8.
    Cleland, J. L., Duenas, E. T., Park, A., Daugherty, A., Kahn, J., Kowalski, J., Cuthbertson, A. 2001Development of poly-(d,l-lactide–coglycolide) microsphere formulations containing recombinant human vascular endothelial growth factor to promote local angiogenesisJ. Control. Release721324CrossRefPubMedGoogle Scholar
  9. 9.
    Amsden, B., Cheng, Y. L. 1995A generic protein delivery system based on osmotically rupturable monolithsJ. Control. Release3399105CrossRefGoogle Scholar
  10. 10.
    Carelli, G., Colo, G., Guerrini, C., Nannipieri, E. 1989Drug release from silicone elastomer through controlled polymer cracking: An extension to macromolecular drugsInt. J. Pharm.50181188CrossRefGoogle Scholar
  11. 11.
    Colo, G. 1992Controlled drug release from implantable matrices based on hydrophobic polymersBiomaterials13850856CrossRefPubMedGoogle Scholar
  12. 12.
    Heller, J., Baker, R. W. 1976System for Delivering Agent to Environment of Use Over Prolonged TimeAlza CorporationUSAGoogle Scholar
  13. 13.
    Amsden, B. G., Cheng, Y. L. 1994Enhanced fraction releasable above percolation—threshold from monoliths containing osmotic excipientsJ. Control. Release312132CrossRefGoogle Scholar
  14. 14.
    Amsden, B. G., Cheng, Y. L., Goosen, M. F. A. 1994A mechanistic study of the release of osmotic agents from polymeric monolithsJ. Control. Release304556CrossRefGoogle Scholar
  15. 15.
    Amsden, B. 2003A model for osmotic pressure driven release from cylindrical rubbery polymer matricesJ. Control. Release93249258CrossRefPubMedGoogle Scholar
  16. 16.
    Gu, F., Younes, H. M., El-Kadi, A. O., Neufeld, R. J., Amsden, B. G. 2005Sustained interferon-gamma delivery from a photocrosslinked biodegradable elastomerJ. Control. Release102607617CrossRefPubMedGoogle Scholar
  17. 17.
    Amsden, B. G., Misra, G., Gu, F., Younes, H. M. 2004Synthesis and characterization of a photo-cross-linked biodegradable elastomerBiomacromolecules524792486CrossRefPubMedGoogle Scholar
  18. 18.
    Ferrara, N., Davis-Smyth, T. 1997The biology of vascular endothelial growth factorEndocr. Rev.18425CrossRefPubMedGoogle Scholar
  19. 19.
    Taniguchi, T., Matsui, H., Fujita, T., Takaoka, C., Kashima, N., Yoshimoto, R., Hamuro, J. 1983Structure and expression of a cloned cdna for human interleukin-2Nature302305310CrossRefPubMedGoogle Scholar
  20. 20.
    Hughes, C. G., Biswas, S. S., Yin, B. L., Baklanov, O. V., DeGrado, T. R., Coleman, R. E., Donovan, C. L., Lowe, J. E., Landolfo, K. P., Annex, B. H. 1999Intramyocardial but not intravenous vascular endothelial growth factor improves regional perfusion in hibernating porcine myocardiumCirculation100476Google Scholar
  21. 21.
    Ferrara, N., Gerber, H. P., LeCouter, J. 2003The biology of VEGF and its receptorsNat. Med.9669676CrossRefPubMedGoogle Scholar
  22. 22.
    King, T. W., Patrick, C. W. 2000Development and in vitro characterization of vascular endothelial growth factor (VEGF)-loaded poly(dl-lactic-co-glycolic acid)/poly(ethylene glycol) microspheres using a solid encapsulation/single emulsion/solvent extraction techniqueJ. Biomed. Materi. Res.51383390CrossRefGoogle Scholar
  23. 23.
    Kurdikar, D. L., Peppas, N. A. 1994Method of determination of initiator efficiency: application to UV polymerization using 2, 2-dimethoxy-2-phenylacetophenoneMacromolecules27733738CrossRefGoogle Scholar
  24. 24.
    Richardson, T. P., Peters, M. C., Ennett, A. B., Mooney, D. J. 2001Polymeric system for dual growth factor deliveryNat. Biotechnol.1910291034CrossRefPubMedGoogle Scholar
  25. 25.
    Gu, F., Amsden, B., Neufeld, R. 2004Sustained delivery of vascular endothelial growth factor with alginate beadsJ. Control. Release96463472CrossRefPubMedGoogle Scholar
  26. 26.
    Kavanagh, C. A., Gorelova, T. A., Selezneva, I. I., Rochev, Y. A., Dawson, K. A., Gallagher, W. M., Gorelov, A. V., Keenan, A. K. 2005Poly(N-isopropy acrylamide) copolymer films as vehicles for the sustained delivery of proteins to vascular endothelial cellsJ. Biomed. Materi. Res. A72A2535CrossRefGoogle Scholar
  27. 27.
    Parmiani, G., Rivoltini, L., Andreola, G., Carrabba, M. 2000Cytokines in cancer therapyImmunol. Lett.744144CrossRefPubMedGoogle Scholar
  28. 28.
    Pardoll, D. M. 1995Paracrine cytokine adjuvants in cancer-immunotherapyAnnu. Rev. Immunol.13399415CrossRefPubMedGoogle Scholar
  29. 29.
    Bos, G. W., Jacobs, J. J. L., Koten, J. W., Tomme, S., Veldhuis, T., Nostrum, C. F., Den Otter, W., Hennink, W. E. 2004In situ crosslinked biodegradable hydrogels loaded with IL-2 are effective tools for local IL-2 therapyEur. J. Pharm. Sci.21 561567CrossRefPubMedGoogle Scholar
  30. 30.
    Pellequer, Y., Ollivon, M., Barratt, G. 2004Formulation of liposomes associated with recombinant interleukin-2: effect on interleukin-2 activityBiomed. Pharmacother.58162167CrossRefPubMedGoogle Scholar
  31. 31.
    Backer, M. V., Aloise, R., Przekop, K., Stoletov, K., Backer, J. M. 2002Molecular vehicles for targeted drug deliveryBioconjug. Chem.13462467CrossRefPubMedGoogle Scholar
  32. 32.
    Thomas, T. T., Kohane, D. S., Wang, A., Langer, R. 2004Mircoparticlulate formulations for the controlled release of interleukin-2J. Pharm. Sci.9311001109CrossRefPubMedGoogle Scholar
  33. 33.
    Rhines, L. D., Sampath, P., DiMeco, F., Lawson, H. C., Tyler, B. M., Hanes, J., Olivi, A., Brem, H. 2003Local immunotherapy with interleukin-2 delivered from biodegradable polymer microspheres combined with interstitial chemotherapy: a novel treatment for experimental malignant gliomaNeurosurgery52872879CrossRefPubMedGoogle Scholar
  34. 34.
    Atkins, T. W., Mccallion, R. L., Tighe, B. J. 1994The incorporation and release of glucose-oxidase and interleukin-2 from a bead formed macroporous hydrophilic polymer matrixJ. Biomater. Sci., Polym. Ed.6651659Google Scholar
  35. 35.
    Ilan, N., Tucker, A., Madri, J. A. 2003Vascular endothelial growth factor expression, beta-catenin tyrosine phosphorylation, and endothelial proliferative behavior: a pathway for transformation?Lab. Invest.8311051115CrossRefPubMedGoogle Scholar
  36. 36.
    Gillis, S., Ferm, M. M., Ou, W., Smith, K. A. 1978T-Cell Growth-Factor—Parameters of production and a quantitative microassay for activityJ. Immunol.12020272032PubMedGoogle Scholar
  37. 37.
    Muller, Y. A., Christinger, H. W., Keyt, B. A., Vos, A. M. 1997The crystal structure of vascular endothelial growth factor (VEGF) refined to 1.93 A resolution: multiple copy flexibility and receptor bindingStructure513251338CrossRefPubMedGoogle Scholar
  38. 38.
    Potgens, A. J., Lubsen, N. H., Altena, M. C., Vermeulen, R., Bakker, A., Schoenmakers, J. G., Ruiter, D. J., Waal, R. M. 1994Covalent dimerization of vascular permeability factor/vascular endothelial growth factor is essential for its biological activity. Evidence from Cys to Ser mutationsJ. Biol. Chem2693287932885PubMedGoogle Scholar
  39. 39.
    Weir, M. P., Chaplin, M. A., Wallace, D. M., Dykes, C. W., Hobden, A. N. 1988Structure activity relationships of recombinant human interleukin-2Biochemistry-Us2768836892CrossRefGoogle Scholar
  40. 40.
    Goolcharran, C., Stauffer, L. L., Cleland, J. L., Borchardt, R. T. 2000The effects of a histidine residue on the c-terminal side of an asparaginyl residue on the rate of deamidation using model pentapeptidesJ. Pharm. Sci.89818825CrossRefPubMedGoogle Scholar
  41. 41.
    Goolcharran, C., Cleland, J. L., Keck, R., Jones, A. J. S., Borchardt, R. T. 2000Comparison of the rates of deamidation, diketopiperazine formation, and oxidation in recombinant human vascular endothelial growth factor and model peptidesAAPS PharmSci216CrossRefGoogle Scholar
  42. 42.
    Sharma, A., Harper, C. M., Hammer, L., Nair, R. E., Mathiowitz, E., Egilmez, N. K. 2004Characterization of cytokine-encapsulated controlled-release microsphere adjuvantsCancer Biother. Radiol.19764769CrossRefGoogle Scholar
  43. 43.
    Henry, T. D., Rocha-Singh, K., Isner, J. M., Kereiakes, D. J., Giordano, F. J., Simons, M., Losordo, D. W., Hendel, R. C., Bonow, R. O., Eppler, S. M., Zioncheck, T. F., Holmgren, E. B., McCluskey, E. R. 2001Intracoronary administration of recombinant human vascular endothelial growth factor to patients with coronary artery diseaseAm. Heart J.142872880CrossRefPubMedGoogle Scholar
  44. 44.
    Fountoulakis, M., Juranville, J. F., Stuber, D., Weibel, E. K., Garotta, G. 1990Purification and biochemical characterization of a soluble human interferon gamma receptor expressed in Escherichia coliJ. Biol. Chem.2651326813275PubMedGoogle Scholar
  45. 45.
    Bocci, V. 1992Physicochemical and biologic properties of interferons and their potential uses in drug delivery systemsCrit. Rev. Ther. Drug Carr. Syst.991133Google Scholar
  46. 46.
    The International Chronic Granulomatous Disease cooperative Study Group1991A controlled trial of interferon gamma to prevent infection in chronic granulomatous diseaseN. Engl. J. Med.324509516CrossRefGoogle Scholar
  47. 47.
    Smith, K. A. 1988Interleukin-2—inception, impact, and implicationsScience24011691176PubMedGoogle Scholar
  48. 48.
    Konrad, M. W., Hemstreet, G., Hersh, E. M., Mansell, P. W. A., Mertelsmann, R., Kolitz, J. E., Bradley, E. C. 1990Pharmacokinetics of recombinant interleukin-2 in humansCancer Res.5020092017PubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2006

Authors and Affiliations

  1. 1.Department of Chemical EngineeringQueen's UniversityKingstonCanada

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