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Poly(Lactide-Co-Glycolide) Microsphere Formulations of Darbepoetin Alfa: Spray Drying Is an Alternative to Encapsulation by Spray-Freeze Drying

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Abstract

Purpose. The purpose of this work was to evaluate spray-freeze drying and spray drying processes for encapsulation of darbepoetin alfa (NESP, Aranesp®).

Methods. Darbepoetin alfa was encapsulated in poly(lactide-co-glycolide) by spray-freeze drying and by spray drying. Integrity was evaluated by size-exclusion chromatography and Western blot. Physical properties and in vitro release kinetics were characterized. Pharmacokinetics and pharmacodynamics were evaluated in nude rats.

Results. Microspheres produced by spray drying were larger than those produced by spray-freeze drying (69 μm vs. 29 μm). Postencapsulation integrity was excellent for both processes, with <2% dimer by size-exclusion chromatography. In vitro release profiles were similar, with low burst (<25%) and low cumulative protein recovery at 4weeks (≤30%), after which time covalent dimer (≤6.5%) and high molecular weight aggregates (≤2.3%) were recovered by denaturing extraction. After a single injection, darbepoetin alfa was detected in serum through 4 weeks for all microsphere formulations tested in vivo, although relative bioavailability was higher for spray-freeze drying (28%) compared with spray drying (21%; p = 0.08) as were yields (73-82% vs. 34-57%, respectively). For both processes hemoglobin was elevated for 7 weeks, over twice as long as unencapsulated drug.

Conclusions. Spray drying, conducted at pilot scale with commercial equipment, is comparable to spray-freeze drying for encapsulation of darbepoetin alfa.

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References

  1. S. D. Putney and P. A. Burke. Improving protein therapeutics with sustained-release formulations. Nat. Biotechnol. 16:153-157 (1998).

    Google Scholar 

  2. S. P. Schwendeman, M. Cardomone, M. R. Brandon, A. Klibanov, and R. Langer. Stability of proteins and their delivery from biodegradable polymer microspheres. In S. Cohen and H. Bernstein (eds.), Microparticulate Systems for the Delivery of Proteins and Vaccines, Marcel Dekker, New York, 1996 pp. 1-49.

    Google Scholar 

  3. P. Herbert, K. Murphy, O. Johnson, N. Dong, W. Jaworowicz, M. A. Tracy, J. L. Cleland, and S. D. Putney. A large-scale process to produce microencapsulated proteins. Pharm. Res. 15:357-361 (1998).

    Google Scholar 

  4. P. A. Burke. Controlled release protein therapeutics: Effects of process and formulation on stability. In D. L. Wise (ed.), Handbook of Pharmaceutical Controlled Release Technology, Marcel Dekker, New York, 2000, pp. 661-692.

    Google Scholar 

  5. J. L. Cleland, O. L. Johnson, S. Putney, and A. J. S. Jones. Recombinant human growth hormone poly(lactic-co-glycolic acid) microsphere formulation development. Adv. Drug Deliv. Rev. 28:71-84 (1997).

    Google Scholar 

  6. P. Giunchedi and U. Conte. Spray-drying as a preparation method of microparticulate drug delivery systems: an overview. S T P Pharma Sci. 5:276-290 (1995).

    Google Scholar 

  7. F. X. Lacasse, P. Hildgen, J. Perodin, E. Escher, N. C. Phillips, and J. N. McMullen. Improved activity of a new angiotensin receptor antagonist by an injectable spray-dried polymer microsphere preparation. Pharm. Res. 14:887-891 (1997).

    Google Scholar 

  8. T. Kissel, Z. Brich, S. Bantle, I. Lancranjan, F. Nimmerfall, and P. Vit. Parenteral depot-systems on the basis of biodegradable polyesters. J. Control. Release 16:27-42 (1991).

    Google Scholar 

  9. K. Masters. Spray Drying in Practice, SprayDryConsult International ApS, Charlottenlund, Denmark, 2002.

    Google Scholar 

  10. P. Giunchedi, B. Conti, I. Genta, U. Conte, and G. Puglisi. Emulsion spray-drying for the preparation of albumin-loaded PLGA microspheres. Drug Devel. Ind. Pharm. 27:745-750 (2001).

    Google Scholar 

  11. B. Bittner and T. Kissel. Ultrasonic atomization for spray drying: a versatile technique for the preparation of protein loaded biodegradable microspheres. J. Microencap. 16:325-341 (1999).

    Google Scholar 

  12. B. Baras, M. A. Benoit, and J. Gillard. Parameters influencing the antigen release from spray-dried poly (DL-lactide) microparticles. Int. J. Pharm. 200:133-145 (2000).

    Google Scholar 

  13. B. Baras, M. A. Benoit, G.-O. Poulain, A. M. Schacht, A. Capron, J. Gillard, and G. Riveau. Vaccine properties of antigens entrapped in microparticles produced by spray-drying technique and using various polyester polymers. Vaccine 18:1495-1505 (2000).

    Google Scholar 

  14. B. Bittner, M. Morlock, H. Koll, G. Winter, and T. Kissel. Recombinant human erythropoietin (rhEPO) loaded poly(lactide-co-glycolide) microspheres: influence of the encapsulation technique and polymer purity on microsphere characteristics. Eur. J. Pharm. Biopharm. 45:295-305 (1998).

    Google Scholar 

  15. C. Perez, I. J. Castellanos, H. R. Costantino, W. Al-Azzam, and K. Griebenow. Recent trends in stabilizing protein structure upon encapsulation and release from bioerodible polymers. J. Pharm. Pharmacol. 54:301-313 (2002).

    Google Scholar 

  16. H. T. Wang, H. Palmer, R. J. Linhardt, D. R. Flanagan, and E. Schmitt. Degradation of poly(ester) microspheres. Biomaterials 11:679-685 (1990).

    Google Scholar 

  17. E. Mathiowitz, H. Bernstein, S. Giannos, P. Dor, T. Turek, and R. Langer. Polyanhydride microspheres. IV. Morphology and characterization of systems made by spray drying. J. Appl. Polym. Sci. 45:125-134 (1992).

    Google Scholar 

  18. S. B. Krantz. Erythropoietin. Blood 77:419-434 (1991).

    Google Scholar 

  19. J. C. Egrie and J. K. Browne. Development and characterization of novel erythropoiesis stimulating protein (NESP). Br. J. Cancer 84(Supp 1):3-10 (2001).

    Google Scholar 

  20. X. C. Nguyen, J. D. Herberger, and P. A. Burke. Protein powders for encapsulation: a comparison of spray-freeze drying and spray drying of darbepoetin alfa. Pharm. Res. 21:507-514 (2004).

    Google Scholar 

  21. O. L. Johnson, W. Jaworowicz, J. L. Cleland, L. Bailey, M. Charnis, E. Duenas, C. C. Wu, D. Shepard, S. Magil, T. Last, A. J. S. Jones, and S. D. Putney. The stabilization and encapsulation of human growth-hormone into biodegradable microspheres. Pharm. Res. 14:730-735 (1997).

    Google Scholar 

  22. J. Herberger, K. Murphy, L. Munyakazi, J. Cordia, and E. Westhaus. Carbon dioxide extraction of residual solvents in poly(lactide-co-glycolide) microparticles. J. Control. Rel. 90:181-195 (2003).

    Google Scholar 

  23. A. G. Hausberger and P. P. DeLuca. Characterization of biodegradable poly(D,L-lactide-co-glycolide) polymers and microspheres. J. Pharm. Biomed. Anal. 13:747-760 (1995).

    Google Scholar 

  24. U. S. Pharmacopeaia and National Formulary, United States Pharmacopeial Convention, Inc., Rockville, MD, 1999.

  25. X. M. Lam, E. T. Duenas, A. L. Daugherty, N. Levin, and J. L. Cleland. Sustained release of recombinant human insulin-like growth factor-I for treatment of diabetes. J. Control. Rel. 67:281-292 (2000).

    Google Scholar 

  26. Y. Ogawa. Injectable microcapsules prepared with biodegradable poly(alpha-hydroxy) acids for prolonged release of drugs. J. Biomater. Sci. Polym. Ed. 8:391-409 (1997).

    Google Scholar 

  27. N. Clarke, K. O'Connor, and Z. Ramtoola. Influence of formulation variables on the morphology of biodegradable microparticles prepared by spray drying. Drug Devel. Ind. Pharm. 24:169-174 (1998).

    Google Scholar 

  28. M. Morlock, H. Koll, G. Winter, and T. Kissel. Microencapsulation of rh-erythropoietin, using biodegradable poly(d,l-lactide-co-glycolide): protein stability and the effects of stabilizing excipients. Eur. J. Pharm. Biopharm. 43:29-36 (1997).

    Google Scholar 

  29. M. A. Tracy, K. L. Ward, L. Firouzabadian, Y. Wang, N. Dong, R. Qian, and Y. Zhang. Factors affecting the degradation rate of poly(lactide-co-glycolide) microspheres in vivo and in vitro. Biomaterials 20:1057-1062 (1999).

    Google Scholar 

  30. T. Heya, H. Okada, Y. Ogawa, and H. Toguchi. In vitro and in vivo evaluation of thyrotropin releasing hormone release from co-poly(dl-lactic/glycolic acid) microspheres. J. Pharm. Sci. 83:636-640 (1994).

    Google Scholar 

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Authors and Affiliations

  1. One Amgen Center, Pharmaceutics and Drug Delivery, Amgen, Inc, Drive Thousand Oaks, California, 93120

    Paul A. Burke, Lisa A. Klumb & Xichdao C. Nguyen

  2. Process Development, Amgen, Inc. One Amgen Center, Drive Thousand Oaks, California, 93120

    John D. Herberger

  3. Pharmacokinetics and Drug Metabolism, Amgen, Inc. One Amgen Center, Drive Thousand Oaks, California, 93120

    Roy A. Harrell & Monica Zordich

Authors
  1. Paul A. Burke
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  2. Lisa A. Klumb
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  5. Roy A. Harrell
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Correspondence to Paul A. Burke.

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Burke, P.A., Klumb, L.A., Herberger, J.D. et al. Poly(Lactide-Co-Glycolide) Microsphere Formulations of Darbepoetin Alfa: Spray Drying Is an Alternative to Encapsulation by Spray-Freeze Drying. Pharm Res 21, 500–506 (2004). https://doi.org/10.1023/B:PHAM.0000019305.79599.a5

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