Pharmaceutical

Experimental
Design
Gareth A. Lewis
Synthelabo Recherche
Chilly Mazarin, France

Didier Mathieu
University of the Mediterranean
Marseille, France

Roger Phan-Tan-Luu
University of Law, Economics, and Science of Aix-Marseille
Marseille, France

MARCEL

MARCEL DEKKER, INC. NEW YORK • BASEL

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 DRUGS AND THE PHARMACEUTICAL SCIENCES

Executive Editor
James Swarbrick
AAI, Inc.
Wilmington, North Carolina

Advisory Board

Larry L. Augsburger David E. Nichols

University of Maryland Purdue University
Baltimore, Maryland West Lafayette, Indiana

Douwe D. Breimer Stephen G. Schulman

Gorlaeus Laboratories University of Florida
Leiden, The Netherlands Gainesville, Florida

Trevor M. Jones Jerome P. Skelly

The Association of the Copley Pharmaceutical, Inc.
British Pharmaceutical Industry Canton, Massachusetts
London, United Kingdom

Hans E. Junginger Felix Theeuwes

Leiden/Amsterdam Center Alza Corporation
for Drug Research Palo Alto, California
Leiden, The Netherlands

Vincent H. L. Lee Geoffrey T. Tucker

University of Southern California University of Sheffield
Los Angeles, California Royal Hallamshire Hospital
Sheffield, United Kingdom

Peter G. Welling
Institut de Recherche Jouveinal
Fresnes, France

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Copyright n 1999 by Marcel Dekker, Inc. All Rights Reserved.

 DRUGS AND THE PHARMACEUTICAL SCIENCES

A Series of Textbooks and Monographs

1. Pharmacokinetics, Milo Gibaldi and Donald Perrier

2. Good Manufacturing Practices for Pharmaceuticals: A Plan for Total
Quality Control, Sidney H. W/l/ig, Murray M. Tuckerman, and William
S, Hitch ings 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 O.
Cooney
10. Concepts in Drug Metabolism (in two parts), edited by Peter Jenner
and Bernard Testa
11. Pharmaceutical Analysis: Modern Methods (in two parts), edited by
James W. Munson
12. Techniques of Solubilization of Drugs, edited by Samuel H. Yalkow-
sky
13. Orphan Drugs, edited by Fred E. Karch
14. Novel Drug Delivery Systems: Fundamentals, Developmental Con-
cepts, Biomedical Assessments, Yie W. Chien
15. Pharmacokinetics: Second Edition, 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. Dermatological Formulations: Percutaneous Absorption, Brian W.
Barry
19. The Clinical Research Process in the Pharmaceutical Industry, edited
by Gary M. Ma to ren
20. Microencapsulation and Related Drug Processes, Patrick B. Deasy
21. Drugs and Nutrients: The Interactive Effects, edited by Daphne A.
Roe and T. Co/in Campbell

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 22. Biotechnology of Industrial Antibiotics, Erick J. Vandamme
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
Bo/ton
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
31. Transdermal Controlled Systemic Medications, edited by Yie W. Chien
32. Drug Delivery Devices: Fundamentals and Applications, edited by
Praveen Ty/e
33. Pharmacokinetics: Regulatory • Industrial • Academic Perspectives,
edited by Peter G. Welling and Francis L. S. Tse
34. Clinical Drug Trials and Tribulations, edited by Alien E. Cato
35. 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
37. Pharmaceutical Pelletization Technology, edited by Isaac Ghebre-
Sellassie
38. Good Laboratory Practice Regulations, edited by Alien F. Hirsch
39. Nasal Systemic Drug Delivery, Yie W. Chien, Kenneth S. E. Su, and
Shyi-Feu Chang
40. Modern Pharmaceutics: Second Edition, Revised and Expanded,
edited by Gilbert S. Banker and Christopher T. Rhodes
41. 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 Bo/ton
45. 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

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Copyright n 1999 by Marcel Dekker, Inc. All Rights Reserved.

 47. HPLC in the Pharmaceutical Industry, edited by Godwin W. Fong and
Stanley K. Lam
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. Coc-
chetto 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 Ex-
panded, 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
61. Pharmaceutical Particulate Carriers: Therapeutic Applications, edited
by A lain 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 Ghebre-Sellassie
66. Colloidal Drug Delivery Systems, edited by Jorg 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
71. Pharmaceutical Powder Compaction Technology, edited by Goran Al-
derborn and Christer Nystrom

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 72. Modern Pharmaceutics: Third Edition, Revised and Expanded, edited
by Gilbert S. Banker and Christopher T. Rhodes
73. Microencapsulation: Methods and Industrial Applications, edited by
Simon Benita
74. Oral Mucosal Drug Delivery, edited by Michael J. Hathbone
75. Clinical Research in Pharmaceutical Development, edited by Barry Bleidt
and Michael Montagne
76. The Drug Development Process: Increasing Efficiency and Cost Effec-
tiveness, 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. Wil-
lig and James R. Stoker
79. Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms: Second
Edition, Revised and Expanded, edited by James W. McGinity
80. Pharmaceutical Statistics: Practical and Clinical Applications, Third
Edition, Sanford Bo/ton
81. Handbook of Pharmaceutical Granulation Technology, edited by Dilip
M. Parikh
82. Biotechnology of Antibiotics: Second Edition, Revised and Expanded,
edited by William R. Strohl
83. Mechanisms of Transdermal Drug Delivery, edited by Russell O. 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: An International Guide to Formula-
tion • Production • 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 Proces-
sing, edited by Joseph F. deSpautz
91. Dermal Absorption and Toxicity Assessment, edited by Michael S.
Roberts and Kenneth A. Walters

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Copyright n 1999 by Marcel Dekker, Inc. All Rights Reserved.

 ADDITIONAL VOLUMES IN PREPARATION

Preparing for PDA Pre-Approval Inspections, edited by Martin D.

Hynes III

Polymorphism in Pharmaceutical Solids, edited by Harry G. Brittain

Pharmaceutical Experimental Design, Gareth A. Lewis, Didier

Mathieu, and Roger Phan-Tan-Luu

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Copyright n 1999 by Marcel Dekker, Inc. All Rights Reserved.

 Preface
Recent years have seen an upsurge in the application of statistical experimental design
methods to pharmaceutical development, as well as to many other fields. These
techniques are no longer the preserve of a few experts and enthusiasts but are becoming
standard tools for the experimenter whose utility is recognised by management. Along
with, or rather preceding, this interest has been a rapid development of the statistical and
computational tools necessary for exploiting the methods. However, their use is not
always optimal and the full range of available methods is often unfamiliar.
Our goal in writing this text is, therefore, to present an integrated approach to
statistical experimental design, covering all important methods while avoiding
mathematical difficulties. The book will be useful not only to the pharmacist or
pharmaceutical scientist developing pharmaceutical dosage forms and wanting the best
and most efficient methods of planning his experimental work, but also to formulation
scientists working in other sectors of the chemical industry. It is not designed primarily for
professional statisticians but these too may find the discussion of pharmaceutical
applications helpful.
Chapter 1 introduces experimental design, describes the book's plan, and
elucidates some key concepts. Chapter 2 describes screening designs for qualitative
factors at different levels, and chapter 3 covers the classic two-level factorial design for
studying the effects of factors and interactions among them. Chapter 4 summarises some
of the mathematical tools required in the rest of the book.
Chapters 5 and 6 deal with designs for predictive models (response surfaces) and
their use in optimization and process validation. Optimization methods are given for
single and for multiple responses. For this we rely mainly on graphical analysis and
desirability, but other methods such as steepest ascent and optimum path are also covered.
The general topic of optimization and validation is continued in chapter 7, on treating
variability, where the "Taguchi" method for quality assurance and possible approaches in
scaling-up and process transfer are discussed.
Chapter 8 covers non-standard designs and the last two chapters are concerned
with mixtures. In chapter 9, the standard mixture design and models are described and
chapter 10 shows how the methods discussed in the rest of the book may be applied to the
more usual pharmaceutical formulation problem where there are severe constraints on the
proportions of the different components. In all of these chapters we indicate how to
choose the optimal design according to the experimenter's objectives, as those most
commonly used — according to the pharmaceutical literature — are by no means optimal
in all situations.

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 However, no book on experimental design of this scope can be considered exhaustive. In
particular, discussion of mathematical and statistical analysis has been kept brief. Designs
for factor studies at more than two levels are not discussed. We do not describe robust
regression methods, nor the analysis of correlations in responses (for example, principle
components analysis), nor the use of partial least squares. Our discussion of variability and
of the "Taguchi" approach will perhaps be considered insufficiently detailed in a few
years. We have confined ourselves to linear (polynomial) models for the most part, but
much interest is starting to be expressed in highly non-linear systems and their analysis by
means of artificial neural networks. The importance of these topics for pharmaceutical
development still remains to be fully assessed.
The words of Qohelet are appropriate here: "And furthermore be admonished, that
of making of books there is no end" (Ecclesiastes 12:13)! He was perhaps unduly
pessimistic as he continued, "and much study is a weariness of the flesh." This was not
our experience, and we trust it will not be that of our readers. In fact, the experience of
putting together this volume on statistical experimental design has proven to be personally
most rewarding. Since we issue from rather different backgrounds, academic and
industrial, chemical and pharmaceutical, French and British, we found that our approaches
contrasted! Reconciling our differences, learning from each other, and integrating our
work, was not always easy, but was invariably stimulating. If we have succeeded in
transmitting to the reader something of what we have learned though this process, we will
be well content.
We would consider all the topics and methods we have covered here likely to be
useful in development, which already takes the book well beyond the two-level factorial
designs equated by many with experimental design itself. This makes for a lot of material
to be digested, but when it is considered that the tools of statistical design may be applied
to such a large part of the development process, we do not feel any apology is called for!
However, the beginner should feel reassured that he does not need such a complete
knowledge before being able to apply these methods to his own work. He should, at least,
read the introductions to each chapter carefully, especially chapters 2, 3, 5, 8 and 9 in
order to decide which situation he is in when a problem arises, whether screening, factor
influence study, modelling, or optimization, and whether there are particular difficulties
and constraints involved. Inevitably he will make some mistakes, as we ourselves have
done, but he will still find that a less-than-ideal choice of design will still give interpretable
results and will be more efficient than the alternative non-design approach.
We wish to express our appreciation to our colleagues, without whose help this
book would not have been written because its origin is in the investigation of
pharmaceutical problems and product development that we have been carrying out
together over many years. In particular we would like to thank Maryvonne Chariot, Jean
Montel, Veronique Andrieu, Frederic Andre, and Gerard Alaux, present and past
colleagues of G.A.L. at Synthelabo, and Michelle Sergent from the University of Aix-
Marseille.

Gareth A. Lewis
Didier Mathieu
Roger Phan-Tan-Luu

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Copyright n 1999 by Marcel Dekker, Inc. All Rights Reserved.

 Contents

Preface

1. Overview
The scope of experimental design

2. Screening
Designs for identifying active factors

3. Factor Influence Studies

Applications of full and fractional factorial designs at 2 levels

4. Statistical and Mathematical Tools

Introduction to multi-linear regression and analysis of variance

5. Response Surface Methodology

Predicting responses within the experimental region

6. Optimization
Pharmaceutical process optimization and validation

1. Variability and Quality

Analysing and minimizing variation

8. Exchange Algorithms
Methods for non-standard designs

9. Mixtures
Introduction to statistical experimental design for formulation

10. Mixtures in a Constrained Region of Interest

Screening, defining the domain, and optimizing formulations

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Copyright n 1999 by Marcel Dekker, Inc. All Rights Reserved.

 Appendix I. A Summary of Matrix Algebra

Appendix II. Experimental Designs and Models for Screening

Appendix III. Designs for Response Surface Modelling

Appendix IV. Choice of Computer Software

Appendix V. Symbols

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Copyright n 1999 by Marcel Dekker, Inc. All Rights Reserved.