Freeze-Drying/lyophilization of Pharmaceutical and Biological Products

Second Edition, Revised and Expanded

Louis Rey
Cabinet d 'Etudes Lausanne, Switzerland

Joan C. May
Center for Biologics Evaluation and Research Food and Drug Administration Rockville, Maryland, U.S.A.

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

Although great care has been taken to provide accurate and current information, neither the author(s) nor the publisher, nor anyone else associated with this publication, shall be liable for any loss, damage, or liability directly or indirectly caused or alleged to be caused by this book. The material contained herein is not intended to provide specic advice or recommendations for any specic situation. Trademark notice: Product or corporate names may be trademarks or registered trademarks and are used only for identication and explanation without intent to infringe. Library of Congress Cataloging-in-Publication Data A catalog record for this book is available from the Library of Congress. ISBN: 0-8247-4868-9 This book is printed on acid-free paper. Headquarters Marcel Dekker, Inc., 270 Madison Avenue, New York, NY 10016, U.S.A. tel: 212-696-9000; fax: 212-685-4540 Distribution and Customer Service Marcel Dekker, Inc., Cimarron Road, Monticello, New York 12701, U.S.A. tel: 800-228-1160; fax: 845-796-1772 Eastern Hemisphere Distribution Marcel Dekker AG, Hutgasse 4, Postfach 812, CH-4001 Basel, Switzerland tel: 41-61-260-6300; fax: 41-61-260-6333 World Wide Web http://www.dekker.com The publisher oers discounts on this book when ordered in bulk quantities. For more information, write to Special Sales/Professional Marketing at the headquarters address above. Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved. Neither this book nor any part may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, microlming, and recording, or by any information storage and retrieval system, without permission in writing from the publisher. Current printing (last digit): 10 9 8 7 6 5 4 3 2 1

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DRUGS AND THE PHARMACEUTICAL SCIENCES

James Swarbrick
PharmaceuTech, Inc. Pinehurst, North Carolina

Advisory Board
Larry L. Augsburger University of Maryland Baltimore, Maryland Harry G. Brittain Center for Pharmaceutical Physics Milford, New Jersey

Jennifer B. Dressman Anthony J. Hickey Johann Wolfgang Goethe-University University of North Carolina School of Pharmacy Frankfurt, Germany Chapel Hill, North Carolina Jeffrey A. Hughes Ajaz Hussain University of Florida College of Pharmacy U.S. Food and Drug Administration Gainesville, Florida Frederick, Maryland Trevor M. Jones Hans E. Junginger The Association of the LeidedAmsterdam Center British Pharmaceutical Industry for Drug Research London, United Kingdom Leiden, The Netherlands Vincent H. L. Lee University of Southern California Los Angeles, California Jerome P. Skelly Alexandria, Virginia Geoffrey T. Tucker University of Sheffield Royal Hallamshire Hospital Sheffield, United Kingdom Stephen G. Schulman University of Florida Gainesville, Florida Elizabeth M. Topp University of Kansas School of Pharmacy Lawrence, Kansas Peter York University of Bradford School of Pharmacy Bradford, United Kingdom

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

DRUGS AND THE PHARMACEUTICALSCIENCES

1. Pharmacokinetics, Milo Gibaldi and Donald Perrier 2. Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control, Sidney H. Willig, Murray M. Tuckerman, and William S. Hitchings IV 3. Microencapsulation, edited by J. R. Nixon 4. Drug Metabolism: Chemical and Biochemical Aspects, Bernard Testa and Peter Jenner 5. New Drugs: Discovery and Development, edited by Alan A. Rubin 6. Sustained and Controlled Release Drug Delivery Systems, edited by Joseph R. Robinson 7. Modern Pharmaceutics, edited by Gilbert S. Banker and Christopher T. Rhodes 8. Prescription Drugs in Short Supply: Case Histories, Michael A. Schwartz 9. Activated Charcoal: Antidotal and Other Medical Uses, David 0. Cooney 10. Concepts in Drug Metabolism (in t w o parts), edited by Peter Jenner and Bernard Testa 11. Pharmaceutical Analysis: Modern Methods (in t w o parts), edited by James W. Munson 12. Techniques of Solubilization of Drugs, edited by Samuel H. Yalkowsky 13. Orphan Drugs, edited by Fred . Karch 14. Novel Drug Delivery Systems: Fundamentals, Developmental Concepts, Biomedical Assessments, Yie W. Chien 15. Phar maco kinet ic s : Second Editio n, Revised and Expanded, Milo Gibaldi and Donald Perrier 16. Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control, Second Edition, Revised and Expanded, Sidney H. Willig, Murray M. Tuckerman, and William S. Hitchings IV 17. Formulation of Veterinary Dosage Forms, edited by Jack Blodinger 18. Der mat oIogic aI FormuIat io ns : Percut aneous Absorption, Brian W. Barry 19. The Clinical Research Process in the Pharmaceutical Industry, edited by Gary M. Matoren 20. Microencapsulation and Related Drug Processes, Patrick B. Deasy 21. Drugs and Nutrients: The Interactive Effects, edited by Daphne A. Roe and T. Colin Campbell 22. Biotechnology of Industrial Antibiotics, rick J. Vandamme

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23. Pharmaceutical Process Validation, edited by Bernard T. Loftus and Robert A. Nash 24. Anticancer and Interferon Agents: Synthesis and Properties, edited by Raphael M. Ottenbrite and George B. Butler 25. Pharmaceutical Statistics: Practical and Clinical Applications, Sanford Bolton 26. Drug Dynamics for Analytical, Clinical, and Biological Chemists, Benjamin J. Gudzinowicz, Burrows T. Younkin, Jr., and Michael J. Gudzinowicz 27. Modern Analysis of Antibiotics, edited by Adjoran Aszalos 28. Solubility and Related Properties, Kenneth C. James 29. Controlled Drug Delivery: Fundamentals and Applications, Second Edition, Revised and Expanded, edited by Joseph R. Robinson and Vincent H. Lee 30. New Drug Approval Process: Clinical and Regulatory Management, edited by Richard A. Guarino 3 1 . Transdermal Controlled Systemic Medications, edited by Yie W. Chien 3 2 . Drug Delivery Devices: Fundamentals and Applications, edited by Praveen Tyle 33. Pharmacokinetics: Regulatory 0 Industrial 0 Academic Perspectives, edited by Peter G. Welling and Francis L. S. Tse 34. Clinical Drug Trials and Tribulations, edited by Allen E. Cat0 3 5 . Transdermal Drug Delivery: Developmental Issues and Research Initiatives, edited by Jonathan Hadgraft and Richard H. Guy 36. Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms, edited by James W. McGinity 3 7 . Pharmaceutical Pelletization Technology, edited by Isaac GhebreSellassie 38. Good Laboratory Practice Regulations, edited by Allen F. Hirsch 39. Nasal Systemic Drug Delivery, Yie W. Chien, Kenneth S. E. Su, and Sh yi-Feu Chang 40. Modern Pharmaceutics: Second Edition, Revised and Expanded, edited by Gilbert S. Banker and Christopher T. Rhodes 4 1 . Specialized Drug Delivery Systems: Manufacturing and Production Technology, edited by Praveen Tyle 42. Topical Drug Delivery Formulations, edited by David W. Osborne and Anton H. Amann 43. Drug Stability: Principles and Practices, Jens T. Carstensen 44. Pharmaceutical Statistics: Practical and Clinical Applications, Second Edition, Revised and Expanded, Sanford Bolton 4 5 . Biodegradable Polymers as Drug Delivery Systems, edited by Mark Chasin and Robert Langer 46. Preclinical Drug Disposition: A Laboratory Handbook, Francis L. S. Tse and James J. Jaffe 47. HPLC in the Pharmaceutical Industry, edited by Godwin W. Fong and Stanley K. Lam

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48. Pharmaceutical Bioequivalence, edited by Peter G. Welling, Francis L. S. Tse, and Shrikant V. Dinghe 49. Pharmaceutical Dissolution Testing, Umesh V. Banakar 50. Novel Drug Delivery Systems: Second Edition, Revised and Expanded, Yie W. Chien 51. Managing the Clinical Drug Development Process, David M. Cocchetto and Ronald V. Nardi 52. Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control, Third Edition, edited by Sidney H. Willig and James R. Stoker 53. Prodrugs: Topical and Ocular Drug Delivery, edited by Kenneth B. Sloan 54. Pharmaceutical Inhalation Aerosol Technology, edited by Anthony J. Hickey 55. Radiopharmaceuticals: Chemistry and Pharmacology, edited by Adrian D. Nunn 56. New Drug Approval Process: Second Edition, Revised and Expanded, edited by Richard A. Guarino 57. Pharmaceutical Process Validation: Second Edition, Revised and Expanded, edited by Ira R. Berry and Robert A. Nash 58. Ophthalmic Drug Delivery Systems, edited by Ashim K. Mitra 59. Pharmaceutical Skin Penetration Enhancement, edited by Kenneth A. Walters and Jonathan Hadgraft 60. Colonic Drug Absorption and Metabolism, edited by Peter R. Bieck 6 1 . Pharmaceutical Particulate Carriers: Therapeutic Applications, edited by Alain Rolland 62. Drug Permeation Enhancement: Theory and Applications, edited by Dean S. Hsieh 63. Glycopeptide Antibiotics, edited by Ramakrishnan Nagarajan 64. Achieving Sterility in Medical and Pharmaceutical Products, Nigel A. Halls 65. Multiparticulate Oral Drug Delivery, edited by Isaac GhebreSellassie 66. Colloidal Drug Delivery Systems, edited by Jlirg Kreuter 67. Pharmacokinetics: Regulatory Industrial Academic Perspectives, Second Edition, edited by Peter G. Welling and Francis L. S. Tse 68. Drug Stability: Principles and Practices, Second Edition, Revised and Expanded, Jens T. Carstensen 69. Good Laboratory Practice Regulations: Second Edition, Revised and Expanded, edited by Sandy Weinberg 70. Physical Characterization of Pharmaceutical Solids, edited by Harry G. Brittain 7 1 . Pharmaceutical Powder Compaction Technology, edited by Goran Alderborn and Christer Nystrom 72. Modern Pharmaceutics: Third Edition, Revised and Expanded, edited by Gilbert S. Banker and Christopher T. Rhodes

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73. Microencapsulation: Methods and Industrial Applications, edited by Simon Benita 74. Oral Mucosal Drug Delivery, edited by Michael J, Rathbone 75. Clinical Research in Pharmaceutical Development, edited by Barry Bleidt and Michael Montagne 7 6 The Drug Development Process: Increasing Efficiency and Cost Effectiveness, edited by Peter G. Welling, Louis Lasagna, and Umesh V. Banakar 77 Microparticulate Systems for the Delivery of Proteins and Vaccines, edited by Smadar Cohen and Howard Bernstein 78 Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control, Fourth Edition, Revised and Expanded, Sidney H. Willg and James R. Stoker 79 Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms: Second Edition, Revised and Expanded, edited by James W. McGinity 8 0 Pharmaceutical Statistics: Practical and Clinical Applications, Third Edition, Sanford Bolton 81 Handbook of Pharmaceutical Granulation Technology, edited by Dilip M. Parikh 82 Biotechnology o f Antibiotics: Second Edition, Revised and Expanded, edited by William R. Strohl 83 Mechanisms of Transdermal Drug Delivery, edited by Russell 0. Potts and Richard H. Guy 84. Pharmaceutical Enzymes, edited by Albert Lauwers and Simon Scharpe 85. Development of Biopharmaceutical Parenteral Dosage Forms, edited by John A. Bontempo 86. Pharmaceutical Project Management, edited by Tony Kennedy 87. Drug Products for Clinical Trials: A n International Guide t o Formulation 0 Production 0 Quality Control, edited by Donald C. Monkhouse and Christopher T. Rhodes 88. Development and Formulation of Veterinary Dosage Forms: Second Edition, Revised and Expanded, edited by Gregory E. Hardee and J. Desmond Baggot 89. Receptor-Based Drug Design, edited by Paul Leff 90. Automation and Validation of Information in Pharmaceutical Processing, edited by Joseph F. deSpautz 91. Dermal Absorption and Toxicity Assessment, edited by Michael S. Roberts and Kenneth A. Walters 92. Pharmaceutical Experimental Design, Gareth A. Lewis, Didier Mathieu, and Roger Phan-Tan-Luu 93. Preparing for FDA Pre-Approval Inspections, edited by Martin D. Hynes Ill 94. Pharmaceutical Excipients: Characterization b y IR, Raman, and NMR Spectroscopy, David E. Bugay and W. Paul Findlay 95. Polymorphism in Pharmaceutical Solids, edited by Harry G. Brittain

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96. Freeze-Drying/Lyophilizationof Pharmaceutical and Biological Products, edited by Louis Rey and Joan C. May 97. Percutaneous Absorption: Drugs-Cosmetics-Mechanisms-Methodology, Third Edition, Revised and Expanded, edited by Robert L. Bronaugh and Howard I. Maibach 98. Bioadhesive Drug Delivery Systems: Fundamentals, Novel Approaches, and Development, edited by Edith Mathiowitz, Donald E. Chickering Ill, and Claus-Michael Lehr 99. Protein Formulation and Delivery, edited by Eugene J. McNally 100. New Drug Approval Process: Third Edition, The Global Challenge, edited by Richard A. Guarino 101. Peptide and Protein Drug Analysis, edited by Ronald E. Reid 102. Transport Processes in Pharmaceutical Systems, edited by Gordon 1. Amidon, Ping I. lee, and Elizabeth M. Topp 103. Excipient Toxicity and Safety, edited by Myra L. Weiner and Lois A. Kotkoskie 104. The Clinical Audit in Pharmaceutical Development, edited by Michael R. Hamrell 105. Pharmaceutical Emulsions and Suspensions, edited by Francoise Nielloud and Gilberte Marti-Mestres 106. Oral Drug Absorption: Prediction and Assessment, edited by Jennifer B. Dressman and Hans Lennernas 107. Drug Stability: Principles and Practices, Third Edition, Revised and Expanded, edited by Jens T. Carstensen and C T. Rhodes . 108. Containment in the Pharmaceutical Industry, edited by James P. Wood 109. Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control from Manufacturer t o Consumer, Fifth Edition, Revised and Expanded, Sidney H. Willig 110. Advanced Pharmaceutical Solids, Jens T. Carstensen 111. Endotoxins: Pyrogens, LAL Testing, and Depyrogenation, Second Edition, Revised and Expanded, Kevin L. Williams 112. Pharmaceutical Process Engineering, Anthony J. Hickey and David Ganderton 113. Pharmacogenomics, edited by Werner Kalow, Urs A. Meyer, and Rachel F. Tyndale 114. Handbook of Drug Screening, edited by Ramakrishna Seethala and Prabhavathi B. Fernandes 115. Drug Targeting Technology: Physical Chemical Biological Methods, edited by Hans Schreier 116. Drug-Drug Interactions, edited by A. David Rodrigues 117. Handbook of Pharmaceutical Analysis, edited by Lena Ohannesian and Anthony J. Streeter 118. Pharmaceutical Process Scale-Up, edited by Michael Levin 119. Dermatological and Transdermal Formulations, edited by Kenneth A. Walters

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120. Clinical Drug Trials and Tribulations: Second Edition, Revised and Expanded, edited by Allen Cato, Lynda Sutton, and Allen Cat0 Ill 121. Modern Pharmaceutics: Fourth Edition, Revised and Expanded, edited by Gilbert S. Banker and Christopher T. Rhodes 122. Surfactants and Polymers in Drug Delivery, Martin Malmsten 123. Transdermal Drug Delivery: Second Edition, Revised and Expanded, edited by Richard H. Guy and Jonathan Hadgraft 1 24. Good Laboratory Practice Regulations: Second Edition, Revised and Expanded, edited by Sandy Weinberg 1 25. Parenteral Quality Control: Sterility, Pyrogen, Particulate, and Package Integrity Testing: Third Edition, Revised and Expanded, Michael J. Akers, Daniel S. Larrimore, and Dana Morton Guazzo 1 26. Modified-Release Drug Delivery Technology, edited by Michael J. Rathbone, Jonathan Hadgraft, and Michael S. Roberts 1 27. Simulation for Designing Clinical Trials: A Pharmacokinetic-Pharmacodynamic Modeling Perspective, edited by Hui C. Kimko and Stephen B. Duffull 1 28. Affinity Capillary Electrophoresis in Pharmaceutics and Biopharmaceutics, edited by Reinhard H. H. Neubert and Hans-Hermann Rii ttinger 1 29. Pharmaceutical Process Validation: An International Third Edition, Revised and Expanded, edited by Robert A , Nash and Alfred H. Wachter 130. Ophthalmic Drug Delivery Systems: Second Edition, Revised and Expanded, edited by Ashim K. Mitra 1 3 1 . Pharmaceutical Gene Delivery Systems, edited by Alain Rolland and Sean M. Sullivan 132. Biomarkers in Clinical Drug Development, edited by John C. Bloom and Robert A. Dean 133. Pharmaceutical Extrusion Technology, edited by Isaac GhebreSellassie and Charles Martin 134. Pharmaceutical Inhalation Aerosol Technology: Second Edition, Revised and Expanded, edited by Anthony J. Hickey 135. Pharmaceutical Statistics: Practical and Clinical Applications, Fourth Edition, Sanford Bolton and Charles Bon 136. Compliance Handbook for Pharmaceuticals, Medical Devices, and Biologics, edited by Carmen Medina 1 37. Freeze-Drying/Lyophilization of Pharmaceutical and Biological Products: Second Edition, Revised and Expanded, edited by Louis Rey and Joan C. May

ADDITIONAL VOLUMES IN PREPARATION

New Drug Approval Process: Fourth Edition, Accelerating Global Registrations, edited by Richard A. Guarino Microbial Contamination Control in Parenteral edited by Kevin L. Williams Manufacturing,

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

Foreword
Like most of todays technological success stories, the history of freezedrying has been pretty much limited to the 20th century. Whereas Altmann (1), in 1890, reported drying frozen tissues to make histological sections, it was not until 1909 that an application to biologicals was rst reported (2). Although the rst patent was led in 1927 (3), there appears to have been little interest in commercial uses until 1935 with a publication by Flosdorf and Mudd (4) that introduced the concept of the cold trap. In the early 1940s Flosdorf et al. (5) in the United States and Greaves (6), working quite independently in England, constructed plants for the large-scale production of dried plasma for wartime use, establishing both the principles and the commercial potential of a new industry. During the subsequent half-century, the potential of freeze-drying captured the imagination of both scientists and industrial engineers, often without full appreciation of the economic limitations imposed by the immutable thermodynamic costs of freezing and the subsequent sublimation of the frozen water. The food industry in particular was attracted by the potential of prolonged room-temperature storage at a time when home freezers were not yet a staple in every kitchen. It is surprising how long it took to recognize the inappropriateness of this demanding technology for use with a low-cost, high-volume product. It is the biological and pharmaceutical industries that have been best able to capitalize on the unique virtues of lyophilization and that have stimulated continuing research into the biophysics of both freezing and freeze-drying, some of which is displayed in the initial chapters of this volume. These are the studies that are progressively converting the pioneering and somewhat brute force demonstrations by Flosdorf and Greaves into modern, more nely tuned procedures, the importance of which cannot be underestimated. At present it is the pharmaceutical industry with its high-cost product that drives the development of this demanding technology that is increasingly delivering great benets to our society.

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

This volume provides clear evidence of the mature state of freeze-drying technology, from the mundane to the sophisticated, all of which are essential to a quality product. And, paradoxically, it is the potential for a high-quality product that will stimulate eorts for still higher quality. The more faithfully the lyophilized product resembles the starting material, the more the focus will fall on the quality of the starting material. The ability to maintain the complex and delicate structural relationships of biologically active compounds during storage at ambient temperatures has been a boon to manufacturers but with an impact well beyond the manufacturing process itself. Analyses of the freeze-drying process are beginning to shift from the physical aspects of freezing and sublimation to a more sophisticated examination of the eects of the process on the chemical structures and the biological properties of the products themselves. It is here that the technology of freeze-drying may ultimately make its greatest contribution. Protein function and protein conformation are inexorably linked, and the forces that maintain functional conformations can be substantially and often irreversibly altered by cold and by dehydration (7,8). As those in the pharmaceutical industry look more closely at how these forces may be altering the structure and jeopardizing the function of biologicals during lyophilization, they will inevitably nd that many of these alterations, particularly those induced by cold, are not limited only to the freeze-drying process but are inherent in many of the isolation and purication procedures conducted well before nal processing begins. An active site on a protein may be a very small proportion of the total molecule. Is maintaining the function of the active site good enough? Why is the rest of the molecule there? Does it make an unrecognized contribution and should we worry about its integrity? The human body is exquisitely designed to reject malformed and altered proteins. Will denaturation of inactive portions of a protein alter its physiological function? Can such denaturation be responsible for unrelated side eects? The resolution of technical concerns such as freezing rates, drying temperatures, and product solubility will permit a more critical reassessment of product quality. As the technology of lyophilization is perfected, it is creating an environment in which attention will be increasingly focused on the stability of product at the molecular level, not just after or even during lyophilization but throughout the entire manufacturing process. The benets to society of safe and more eective pharmaceuticals are indisputable and for those of us

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

who have participated in the development of freeze-drying technology, it is a privilege to have been part of that history. Harold T. Meryman, M.D. Founding Member and Past-President The Society of Cryobiology Rockville, Maryland, U.S.A.

REFERENCES
1. 2. 3. 4. 5. 6. 7. R Altmann. Die Elementarorganismen und ihre Beziehungen zu den Lellen. Veit and Co., Leipzig, 1890. L Shackell. Amer. J. Physiol. 24:325, 1909. HL Tival. U.S. Patent No. 1,630,985, 1927 (and 1932, No. RE 18,364). EW Flosdorf and S Mudd. J. Immunol. 29:389, 1935. EW Flosdorf, F Stokes, and S Mudd. J. Amer. Med. Ass. 115:1095, 1940. RIN Greaves. The Preservation of Proteins by Drying, H.M.S.O., London, 1946. JF Carpenter, BS Chang. Lyophilization of protein pharmaceuticals. Biotechnology and Biopharmaceutical Manufacturing, Processing and Preservation. Edited by KE Avis and VL Wu, Volume 2, 199264, 1996. LI Tsonev and AG Hirsh. Fluorescence ratio intrinsic basic states analysis: a novel approach to monitor and analyze protein unfolding by uorescence. J. of Biochemical and Biophysical Methods 45, Issue 1, 121.

8.

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Foreword
When I rst became aware of the freezing process, some 30 years ago, the process appeared to me to be rather simple and straightforward. At that time I was employed by a major manufacturer of freeze-dryers but working in the eld of microelectronics. Given my background in physical chemistry, I found myself becoming increasingly involved in the eld of freeze-drying. I must admit I was surprised and somewhat puzzled to learn that some of those using this process were experiencing diculties. Many formulations of lyophilized products were made isotonic and contained a host of other recipients such as bulking compounds, cryoprotectants, surfactants, and pH modiers. It was then that I started to read publications concerning the freeze-drying process, particularly those that oered any explanation regarding why some freeze-drying processes were not successful. It was at that time that I realized that the freeze-drying process was more complex than my rst impression. That viewpoint has not been altered and perhaps has only been reinforced as the years have passed. However, a great deal of research has been and continues to be done in this eld to enhance our understanding. The actual number of factors we should take into account with a freeze-drying or lyophilization process I simply never stopped to count. But I am certain that they will exceed the 10 ngers on our hands. While not wishing to examine every possible aspect of this process, let me just share with the reader just three areas that I feel to be of major concern. The rst and foremost is the thermal properties of the formulation, without which one is reduced to process development by trial and error. With knowledge of the thermal properties, one is able to quickly develop and validate a lyophilization process. Without such knowledge one has no reference point on which to rely should a change occur in the properties of the formulation. Knowledge of the thermal properties is paramount to the development of a lyophilization process. The stability of a lyophilized product will be dependent on the residual moisture content. The industry is certainly very much in need of a means

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

of determining the residual moisture in a product that is both nondestructive and noninvasive. The method should determine the moisture without destroying the dried product, nor should the measurement cause any changes in the product properties such as a loss in activity. Finally, although the equipment should only provide a safe environment for the product and the necessary operating parameters for the lyophilization process, dierences in freeze-drying equipment can aect the process. One should be aware of such dierences, especially when transferring a process from one dryer to another. So in any discussion of the lyophilization process, the freeze-drying equipment should not be overlooked. It is the intent of this Foreword to provide the reader with an appreciation that considerable eorts are being made to enhance our knowledge of the lyophilization process and its associated instrumentation and equipment. While admittedly we will need more information to complete our knowledge of this process, the advances described in this book will take us closer to achieving this nal objective. Thomas A. Jennings, Ph.D. President The International Society for LyophilizationFreeze-Drying, Inc. Bala Cynwyd, Pennsylvania, U.S.A.

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Preface
In the middle of the 1950s, when I was actively engaged in low-temperature preservation of living tissues and organs, and tissue banking, I discovered with surprise that in 1902, in St. Petersburg, the participants of the International Congress on Paleontology were given mammoth meat at the banquet. That curiosity actually came from Iakoutia, where a whole frozen mammoth suddenly appeared in the collapsed bank of a Siberian river. Apparently this body had been stored there in the permafrost for 15,000 years and the wolves still found it palatable, as did the conference participants. I learned then that this discovery was not uncommon and that, from time to time, well-preserved mammoth were found in Northern Siberia. Quite excited by this news I started to investigate whether I could get a sample of that unique material. Numerous requests made at all levels of the administration were fruitless and I had almost forgotten the issue when, in the late 1960s, I received in Switzerland a big box from the USSR Academy of Sciences andwhat a surpriseinside there was a big mammoth steak with its fur still attached to the skin. Instead of being frozen it was perfectly freeze-dried and, of course, naturally so. Apparently the big animal had broken its backbone falling into a crevasse and was buried under snow, almost immediately and relatively close to the surface, where it sublimed for millennia, which kept its anatomical features almost intact. We made many scientic investigations of this sample and obtained excellent electron micrographs of the dried muscles. Less successful, by far, was the stew that the Nestle cooks managed to prepare with it : it denitely was no delicacy! Much more recently, when opening a microbiological conference in Morocco, I came across in my preparation work, on several interesting papers dealing with lithopanspermia, the fantastic ride of living cells in suspended animation, dashing throughout outer space on a rocks back. Quite certainly freeze-dried, they traveled there for maybe millions of years before being captured by the earths gravity eld and falling into the depths

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of the ocean, whereit is claimed by some scientiststhey seeded life on our planet. Freeze-drying, as a natural phenomenon, was again in the limelight. A scientic curiosity for almost 40 years after the publication of Bordas and dArsonval in 1906, freeze-drying, later called lyophilization by Earl Flosdorf at the time of World War II, has bloomed again within the past 20 years to become an almost unavoidable technology to preserve rare and sensitive biochemicals and drugs. Concomitantly, new challenges appeared in basic and applied engineering elds and required more and more sophisticated approaches. This is the reason that Joan May and I found it useful, if not compulsory, to prepare a second edition of our 1999 book Freeze-Drying/ Lyophilization of Pharmaceutical and Biological Products. Indeed, our understanding of the fundamentals of freeze-drying has been continuously improving in such areas as conned water, annealing, NMR assessment of mobility in dried products, formulation, protein stabilization, and the role of additives. We witness rising interest in some elds which have been long considered as collateral but which are of prime importance today: properties and behavior of glass and elastomeres in the always present container-closer system. In parallel, industrial operations are becoming more diversied and oer numerous dierent problems: scaling up towards production from the laboratory bench throughout the pilot plant, cleaning and environmental concerns, and sterile handling of bulk material with the associated qualication and validation strategies. Moreover, new technologies are starting to develop: the use of co-solvents and irradiation. However, in all cases, an absolute duty of care still remains for the operators to provide both security and quality and keep their outgoing products in line with the international standards, a eld which is quickly expanding. Thus, year after year, lyophilization is becoming a vast, diversied eld for research and development, engineering, and production, still under the close eye of the administration and of the compliance ocers. Therefore, it comes as no surprise that the publisher and the editors decided to prepare this revised and expanded second edition. In so doing, it is a privilege and pleasure for Joan May and I to extend our warm appreciation to all our devoted, competent contributors and acknowledge once more the generous support and professional skill of the sta at Marcel Dekker, Inc. Louis Rey, Ph.D.

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

Contents
Foreword Harold T. Meryman Foreword Thomas A. Jennings Preface Louis Rey Contributors 1 Glimpses into the Realm of Freeze-Drying: Fundamental Issues Louis Rey 2 Structural and Dynamic Properties of Bulk and Conned Water M.-C. Bellissent-Funel and J. Teixeira Mechanisms of Protein Stabilization During Freeze-Drying and Storage: The Relative Importance of Thermodynamic Stabilization and Glassy State Relaxation Dynamics Michael J. Pikal Freezing and Annealing Phenomena in Lyophilization James A. Searles Freezing- and Drying-Induced Perturbations of Protein Structure and Mechanisms of Protein Protection by Stabilizing Additives John F. Carpenter, Ken-ichi Izutsu, and Theodore W. Randolph Molecular Mobility of Freeze-Dried Formulations as Determined by NMR Relaxation, and its Eect on Storage Stability Sumie Yoshioka Formulation Characterization D. Q. Wang

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Practical Aspects of Freeze-Drying of Pharmaceutical and Biological Products Using Non-Aqueous Co-Solvent Systems Dirk L. Teagarden and David S. Baker

Closure and Container Considerations in Lyophilization Frances L. DeGrazio Advanced Glassware for Freeze-Drying Jurgen Thurk and Peter Knaus

10

11

Critical Steps in the Preparation of Elastomeric Closures for Biopharmaceutical Freeze-Dried Products Maninder S. Hora and Sidney N. Wolfe

12

Development of a New Concept for Bulk Freeze-Drying: LYOGUARD Freeze-Dry Packaging Meagan Gassler and Louis Rey

13

Regulatory Control of Freeze-Dried Products: Importance and Evaluation of Residual Moisture Joan C. May

14

Freeze-Drying of Biological Standards Paul Matejtschuk, Michelle Andersen, and Peter Phillips Industrial Freeze-Drying for Pharmaceutical Applications Georg-Wilhelm Oetjen

15

16

Development of Process Data in a Pilot Plant Transferable to Production Hanna Willemer

17

Technical Procedures for Operation of Cleaning-in-Place/Sterilization-in-Place Process for Production Freeze-Drying Equipment Gilles A. Beurel

18

Global Validation of Freeze-Drying Cycle Parameters by Using Integral HFT Systems Gilles A. Beurel

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19

Lyophilizer Qualication: Some Practical Advice Thorsten Fischer

20

Lyophilization Process Validation Christian Bindschaedler A New Development: Irradiation of Freeze-Dried Vaccine and Other Select Biological Products Louis Rey and Joan C. May

21

22

Some Leading Edge Prospects in Lyophilization Louis Rey

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Contributors
Michelle Andersen Standards Division, National Institute for Biological Standards & Control (NIBSC), Hertfordshire, England David S. Baker Pfizer Corporation, Kalamazoo, Michigan, U.S.A.

M.-C. Bellissent-Funel Laboratoire Leon-Brillouin (CEA-CNRS), CEA Saclay, Gif-sur-Yvette, France Gilles A. Beurel S.G.D. Serail, Argenteuil, France Serono Laboratories S.A., Aubonne, Switzerland

Christian Bindschaedler

John F. Carpenter University of Colorado Health Sciences Center, Denver, Colorado, U.S.A. Frances L. DeGrazio Pennsylvania, U.S.A. Thorsten Fischer Meagan Gassler U.S.A. Maninder S. Hora West Pharmaceutical Services, Inc., Lionville,

Aventis Behring GmbH, Marburg, Germany W. L. Gore & Associates, Inc., Elkton, Maryland, Chiron Corporation, Emeryville, California, U.S.A.

Ken-ichi Izutsu University of Colorado Health Sciences Center, Denver, Colorado, U.S.A. Peter Knaus Switzerland Forma Vitrum, Schott Pharmaceutical Packaging, St. Gallen,

Paul Matejtschuk Standards Division, National Institute for Biological Standards & Control (NIBSC), Hertfordshire, England Joan C. May Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, Maryland, U.S.A. Georg-Wilhelm Oetjen Lubeck, Germany

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.

Peter Phillips Standards Division, National Institute for Biological Standards & Control (NIBSC), Hertfordshire, England Michael J. Pikal School of Pharmacy, University of Connecticut, Storrs, Connecticut, U.S.A. Theodore W. Randolph Louis Rey University of Colorado, Boulder, Colorado, U.S.A. Cabinet dEtudes, Lausanne, Switzerland

James A. Searles Global Parenteral Products, Manufacturing Science and Technology, Eli Lilly and Company, Indianapolis, Indiana, U.S.A. Dirk L. Teagarden Pfizer Corporation, Kalamazoo, Michigan, U.S.A.

J. Teixeira Laboratoire Leon-Brillouin (CEA-CNRS), CEA Saclay, Gif-sur-Yvette, France Jurgen Thurk Forma Vitrum, Schott Pharmaceutical Packaging, St. Gallen, Switzerland D. Q. Wang Bayer Corporation, Berkeley, California, U.S.A. Koln, Germany Chiron Corporation, Emeryville, California, U.S.A. National Institute of Health Sciences, Tokyo, Japan

Hanna Willemer Sidney N. Wolfe Sumie Yoshioka

Copyright 2004 by Marcel Dekker, Inc. All Rights Reserved.