The Stability of Insulin in Crystalline and Amorphous Solids: Observation of Greater Stability for the Amorphous Form

Pharmaceutical Research - Tập 14 Số 10 - Trang 1379-1387 - 1997
Pikal, Michael J.1, Rigsbee, Daniel R.1
1Lilly Research Laboratories, Eli Lilly & Co, Indianapolis

Tóm tắt

Purpose. Generalizations based upon behavior of small molecules have established that a crystalline solid is generally much more stable toward chemical degradation than is the amorphous solid. This study examines the validity of this generalization for proteins using biosynthetic human insulin as the model protein. Methods. Amorphous insulin was prepared by freeze drying the supernate from a suspension of zinc insulin crystals adjusted to pH 7.1. Storage stability at 25°C and 40°C were compared for the freeze dried material, the dried suspended crystals, and the starting batch of crystals. Samples were equilibrated at selected relative humidities between zero and 75% to obtain samples at various water contents. Assays for dimer formation were performed by size exclusion HPLC and assays for deamidated product were carried out by reverse phase HPLC. Degradation was found to be linear in square root of time, and the slopes from % degradation vs. square root of time were used to define the rate constants for degradation. Differential scanning calorimetry (DSC) and Fourier-transform infrared spectroscopy (FTIR) were used to characterize the state of the protein in the solids. Results. As expected based upon previous results, the primary degradation pathways involve deamidation at the AsnA21 site and co-valent dimer formation, presumably involving the A-21 site. Contrary to expectations, amorphous insulin is far more stable than crystalline insulin under all conditions investigated. While increasing water content increases the rate of degradation of crystalline insulin, rate constants for degradation in the amorphous solid are essentially independent of water content up to the maximum water content studied (≈15%). Conclusions. Based upon the FTIR and DSC data, both crystalline and amorphous insulin retain some higher order structure when dried, but the secondary structure is significantly perturbed from that characteristic of the native solution state. However, neither DSC nor FTIR data provide a clear interpretation of the difference in stability between the amorphous and crystalline solids. The mechanism responsible for the superior stability of amorphous insulin remains obscure.

Từ khóa


Tài liệu tham khảo

citation_journal_title=J. Pharm. Sci.; citation_author=M. J. Pikal, A. L. Lukes, J. E. Lang, K. Gaines; citation_volume=67; citation_publication_date=1978; citation_pages=767; citation_id=CR1

citation_journal_title=Chem. Pharm. Bull.; citation_author=T. Oguchi, E. Yonemochi, K. Yamamoto, Y. Nakai; citation_volume=37; citation_publication_date=1989; citation_pages=3088-3091; citation_id=CR2

M. J. Pikal. “Freeze Drying of Proteins”, in Peptide and Protein Delivery, 2nd Edition, V. H. L. Lee, Editor, Marcel Dekker, in press.

citation_journal_title=J. Am. Chem. Soc.; citation_author=C. N. Sukenik, J. A. P. Bonapase, N. S. Mandel, R. G. Bergman, P. Lau, G. Wood; citation_volume=97; citation_publication_date=1975; citation_pages=5290-5291; citation_id=CR4

citation_journal_title=Pharm. Res.; citation_author=J. Brange, L. Langkjaer, S. Havelund, A. Volund; citation_volume=9; citation_publication_date=1992; citation_pages=715-726; citation_id=CR5

citation_journal_title=Arch. Biochem. Biophys.; citation_author=S. J. Prestrelski, T. Arakawa, J. F. Carpenter; citation_volume=303; citation_publication_date=1993; citation_pages=465-473; citation_id=CR6

citation_journal_title=Pharm. Res.; citation_author=S. J. Prestrelski, K. A. Pikal, T. Arakawa; citation_volume=12; citation_publication_date=1995; citation_pages=1250-1259; citation_id=CR7

citation_journal_title=Pharm. Res.; citation_author=J. Brange, S. Havelund, P. Hougaard; citation_volume=9; citation_publication_date=1992; citation_pages=727-734; citation_id=CR8

citation_journal_title=Acta Pharm. Nord.; citation_author=J. Brange, O. Hallund, E. Sorensen; citation_volume=4; citation_publication_date=1992; citation_pages=223-232; citation_id=CR9

citation_journal_title=J. Pharm. Sci.; citation_author=R. T. Darrington, B. D. Anderson; citation_volume=84; citation_publication_date=1995; citation_pages=275-282; citation_id=CR10

citation_journal_title=Pharm. Res.; citation_author=R. G. Strickley, B. D. Anderson; citation_volume=13; citation_publication_date=1996; citation_pages=1142-1153; citation_id=CR11

citation_journal_title=J. Pharm. Sci.; citation_author=A. Dong, S. J. Prestrelski, S. D. Allison, J. F. Carpenter; citation_volume=84; citation_publication_date=1995; citation_pages=415-424; citation_id=CR12

citation_journal_title=Biochem.; citation_author=A. Dong, B. Caughey, W. S. Caughey, K. S. Bhat, J. E. Coe; citation_volume=31; citation_publication_date=1992; citation_pages=9364-9370; citation_id=CR13

citation_journal_title=Methods in Enzymology.; citation_author=L. J. Janis, P. M. Kovach, R. M. Riggin, J. K. Towns; citation_volume=271; citation_publication_date=1996; citation_pages=86-113; citation_id=CR14

citation_journal_title=Drug Dev. Ind. Pharm.; citation_author=M. J. Hageman; citation_volume=14; citation_publication_date=1988; citation_pages=2047-2070; citation_id=CR15

citation_journal_title=Pharm. Res.; citation_author=M. J. Pikal, K. M. Dellerman, M. L. Roy, R. M. Riggin; citation_volume=8; citation_publication_date=1991; citation_pages=427-436; citation_id=CR16

citation_journal_title=Develop Biol. Standard.; citation_author=M. J. Pikal, K. Dellerman, M. L. Roy; citation_volume=74; citation_publication_date=1991; citation_pages=323-340; citation_id=CR17

citation_journal_title=J. Pharm. Pharmac.; citation_author=A. T. M. Serajuddin, M. Rosoff, D. Mufson; citation_volume=38; citation_publication_date=1985; citation_pages=219-220; citation_id=CR18

citation_journal_title=J. Phys. Chem.; citation_author=J. L. Green, J. Fan, C. A. Angell; citation_volume=98; citation_publication_date=1994; citation_pages=13780-13790; citation_id=CR19

citation_journal_title=Pharm. Res.; citation_author=B. C. Hancock, S. L. Shamblin, G. Zografi; citation_volume=12; citation_publication_date=1995; citation_pages=799-806; citation_doi=10.1023/A:1016292416526; citation_id=CR20

citation_journal_title=J. Pharm. Sci.; citation_author=L. N. Bell, M. J. Hageman, L. M. Muraoka; citation_volume=84; citation_publication_date=1995; citation_pages=707-712; citation_id=CR21

citation_journal_title=Food Hydrocolloids.; citation_author=I. V. Sochava, O. I. Smirnova; citation_volume=6; citation_publication_date=1993; citation_pages=513-524; citation_id=CR22

M. J. Pikal and D. Rigsbee. The relationship between glass transition temperature and stability of freeze dried human growth hormone formulations. AAPS Pharmaceutical Technology Symposium: Viscoelastic properties of pharmaceutical materials, AAPS National Meeting, Miami, Florida, Nov. 7, 1995.

citation_journal_title=Biochem.; citation_author=J. R. Lepock, K. P. Ritchie, M. C. Kolios, A. M. Rodahl, K. A. Heinz, J. Kruuv; citation_volume=31; citation_publication_date=1992; citation_pages=12706-12712; citation_id=CR24

citation_journal_title=Biophysical J.; citation_author=S. J. Prestrelski, N. Tedeschi, T. Arakawa, J. F. Carpenter; citation_volume=65; citation_publication_date=1993; citation_pages=661-671; citation_id=CR25

citation_journal_title=Biochem.; citation_author=Y. Pocker, S. B. Biswas; citation_volume=19; citation_publication_date=1980; citation_pages=5043-5049; citation_id=CR26

citation_journal_title=Biochim. Biophys. Acta.; citation_author=H. R. Costantino, K. Griebenow, P. Mishra, R. Langer, A. M. Klibanov; citation_volume=1253; citation_publication_date=1995; citation_pages=69-74; citation_id=CR27

citation_journal_title=J. Non-Cryst. Solids.; citation_author=C. A. Angell; citation_volume=102; citation_publication_date=1988; citation_pages=205-221; citation_id=CR28

R. G. Strickley. Stability of human insulin: Effect of water content and added excipients on the degradation of lyophilized insulin at acidic pH. Ph.D. Dissertation, The University of Utah. December, 1995.

Thông tin
Thông tin xuất bản

Pharmaceutical Research

Tập 14 Số 10

1379-1387

Thông tin tác giả