Drug Absorption Studies

Biotechnology: Pharmaceutical Aspects Volume I: Pharmaceutical Profiling in Drug Discovery for Lead Selection R.T. Borchardt, E.H. Kerns, C.A. Lipinski, D.R. Thakker, B. Wang Volume II: Lypophilization of Biopharmaceuticals H.R. Constantino, M.J. Pikal Volume III: Methods for Structural Analysis of Protein Pharmaceuticals W. Jiskoot, D.J.A. Crommelin Volume IV: Optimizing the “Drug-Like” Properties of Leads in Drug Discovery R.T. Borchardt, E.H. Kerns, M.J. Hageman, D.R. Thakker, J.L. Stevens Volume V: Prodrugs: Challenges and Rewards, Parts 1 and 2 V.J. Stella, R.T. Borchardt, M.J. Hageman, R. Oliyai, H. Maag, J.W. Tilley Volume VI: Solvent Systems and Their Selection in Pharmaceutics and Biopharmaceutics P. Augustijns, M.E. Brewster Volume VII: Drug Absorption Studies: In Situ, In Vitro and In Silico Models C. Ehrhardt, K.J. Kim

In Vitro and In Silico Models 123 .Carsten Ehrhardt Editors Kwang-Jin Kim Drug Absorption Studies In Situ.

edu ISBN 978-0-387-74900-6 e-ISBN 978-0-387-74901-3 Library of Congress Control Number 2007937951 c 2008 American Association of Pharmaceutical Scientists All rights reserved. NY 10013. The use in this publication of trade names. USA). service marks. This work may not be translated or copied in whole or in part without the written permission of the publisher (Springer Science+Business Media. even if they are not identified as such. USA Telephone: +1 323 442 1217 Fax: +1 323 442 2611 Email: Kwang.com . Ireland Telephone: +353 1 896 2441 Fax: +353 896 2783 Email: ehrhardc@tcd. 233 Spring Street.Editors Carsten Ehrhardt School of Pharmacy and Pharmaceutical Sciences Trinity College Dublin Panoz Institute. CA 90033. Printed on acid-free paper. LLC. trademarks.ie Kwang-Jin Kim Keck School of Medicine University of Southern California Room HMR-914. Use in connection with any form of information storage and retrieval. and similar terms. 2011 Zonal Avenue Los Angeles.J. computer software. or by similar or dissimilar methodology now known or hereafter developed is forbidden. electronic adaptation. Westland Row Dublin 2. is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. except for brief excerpts in connection with reviews or scholarly analysis. 9 8 7 6 5 4 3 2 1 springer.Kim@usc. New York.

1 Skin Sensitization 1.2 Structure and Function of the Skin 1.2.2.2. Marc Schneider.3.3.1 Anatomical Structure of Human Skin 1.2 General Model Describing Gastrointestinal Absorption 2.Contents Preface Contributors Part 1 Perfused Organ Level/In Situ Techniques 1.2 Biological Activity of the Skin 1.4 Estimating Effective Intestinal Permeability Coefficient Using a Mass Balance Approach 2.3.2.1 In Vivo Studies Using Pharmacodynamic Response 1.3.5 Using Peff to Estimate the Extent of Absorption xv xvii 3 4 4 4 6 6 7 8 9 9 10 11 12 16 18 18 21 23 34 35 37 37 40 41 42 44 v .4.3 Skin Appendages 1.1 Estimating Drug Absorption Trends from Physiochemical Characteristics 2.4. Models for Skin Absorption and Skin Toxicity Testing Ulrich F.2.2 In Vivo Dermatopharmacokinetic Approach 1. and Claus-Michael Lehr 1.2.3.2.2. Javiana Luengo Contreras. Steffi Hansen.1 Introduction 2.2 Skin Irritation and Corrosion 1.4 Perfused Skin Models 1.3.2 Theoretical Models Describing the Gastrointestinal Absorption of Drugs 2.3 In Vivo Dermal Microdialysis 1. Models of the Small Intestine Brendan Griffin and Caitriona O’Driscoll 2.6 In Vitro Skin Penetration Studies 1.4 Testing on Skin Toxicity 1.4. Schaefer.1 Introduction 1.4 Skin Absorption Pathways 1.3 Strategies for Skin Invasion Testing Classified According to Their Resemblance of the In Vivo Situation 1.5 In Vitro Skin Permeation Studies 1.2.3 Skin Phototoxicity 2.3 The Effective Permeability Coefficient 2.

1 Carrier-Mediated Transport 3.2 Intestinal Perfusion with Venous Sampling Models 2.2 In Situ Rat Colon Model for Absorption Evaluation 3.4 Mesenteric Lymph Duct Cannulated Anaesthetised Rat Model 2.2 In Vitro Methods 4.5. Nicolazzo and Barrie C.1 Cannulated Conscious Rat Models 2.3.3.3.2 Passive Transport 3.2.5.4.5 Methods Employed to Assess the Permeability of the Buccal Mucosa 4. Finnin 4.2 Carrier-Mediated Transport 4.3 The Isolated and Vascularly Perfused Intestinal Models 2.3 Single-Pass Perfusion in Conscious Dog/Pig–Loc-I-Gut 2.3.5 Anaesthetised Large Animal Model 2.2 Structure and Environment of the Buccal Mucosa 4.1 In Vivo Methods 4.3.1 Standardisation and Validation Criteria in Peff Determination 2.3.1 Introduction 3. Drug Absorption from the Colon In Situ Hiroaki Yuasa 3.5.3 The Barriers of the Buccal Mucosa 4.4 Concluding Remarks 4.5.5.vi Contents 2.3 In Situ Models 2.4.2 Cannulated Conscious Large Animal Model 2.2.1 Epithelial Organization 4.3.3.1 Intestinal Perfusion Techniques 2.5 Discussion 2.3 Permeability Characteristics of the Rat Colonic Membrane 3.3 Buccal Cell Cultures 4.4 In Vivo Models 2.5.6 Concluding Remarks 46 46 50 53 54 55 55 56 57 59 60 62 63 63 65 77 78 79 81 81 84 86 89 89 90 90 90 91 91 92 92 94 94 94 95 96 97 102 103 .4 Mechanisms Involved in Oral Mucosal Absorption 4.1 Passive Diffusion 4.3 Choice of Animal Species 3. In Vivo and In Vitro Models for Assessing Drug Absorption Across the Buccal Mucosa Joseph A.1 Location of the Permeability Barrier 4.4.4.2 Choice of in Situ Versus in Vivo Models 2.3.3.1 Introduction 4.4.3 Other Permeability Barriers in the Buccal Mucosa 4.2 Organization of the Intercellular Domain 4.4 Single-Pass Perfusion in Conscious Humans—Loc-I-Gut 2.2 Chemical Nature of the Permeability Barrier 4.4.

Cynthia Bosquillon.1.2 Experimental Set-up 6. The Isolated Perfused Lung for Drug Absorption Studies Ann Tronde.1 Introduction 112 113 114 116 117 117 118 119 121 121 121 125 126 127 127 129 135 136 136 137 141 142 143 146 146 147 150 151 154 167 167 .3 Drug Administration to the Lung 6.5.4 Nasal Drug Irritation and Tolerance 5.2.2.1.1 Permeability Studies 5. and Ben Forbes 6.4. Hidalgo 7.1 Principles of the Preparation 6.3 Optimization of Formulation Parameters and Drug Absorption Enhancement Strategies 5.5 Lung Model Selection for Drug Absorption Studies 6.5 Specific Applications of Ex Vivo Models in Nasal Drug Delivery Studies 5.5 Developments in the IPL Part 2 Cellular Level—In Vitro Models of Epithelial and Endothelial Barriers 7.7 Conclusions 6.4.2 Metabolism Studies 5.2 The Isolated Perfused Lung 6. In Vitro Models for Investigations of Buccal Drug Permeation and Metabolism Tanja Obradovic and Ismael J.1.6 Correlation Between Nasal Drug Absorption Models 5.1 Permeability Studies and Characterization of Drug Absorption Pathways 5.4.1.5.4 Drug Absorption in the Lung 6.Contents vii 5.4.4 Specific Applications of in Situ Methods in Nasal Drug Delivery Studies 5.1.1 Introduction 5.2 Advantages of Ex Vivo and in Situ Models for Drug Absorption and Metabolism Studies 5.1 Inhaled Drug Delivery 6.2.3 Optimization of Drug Absorption Enhancement Strategies 5.3 Limitations of Ex Vivo and in Situ Models for Drug Absorption and Metabolism Studies 5. Agu and Michael I.2.5.1 Respiratory Drug Delivery 6.2 Metabolism Studies 5.4 Nasal Drug Irritation and Tolerance 5.3 Drug Administration to the IPL 6. In Situ and Ex Vivo Nasal Models for Preclinical Drug Development Studies Remigius U.2 The Lung 6.5.4 Drug Absorption Studies Using the Isolated Perfused Lung 6.2. Ugwoke 5.

2.1 Introduction 8.3 In Vitro Methods to Assess Intestinal Metabolism 8.2 Physicochemical Characteristics of the Drugs 9.2 The Intestinal Mucosa as a Physical and Biochemical Barrier to Drug Absorption 8.2 Paracellular Route 9.2 In Vitro Models of the Tracheo-Bronchial Epithelium 10.2.2.3 Concluding Remarks 8.4 Concluding Remarks 9.viii Contents 7.2 Cell Culture-Based Models (Caco-2) 8.1.3.1 Membrane-Based Models (PAMPA) 8. and Kwang-Jin Kim 10. Pieter Annaert.2 Physiology of Nasal Mucosa 9. In Vitro Models of the Tracheo-Bronchial Epithelium Carsten Ehrhardt.5.1.3 Cell Line Models 9.2.2. and Patrick Augustijns 8.4 Transport Routes of Nasal Epithelial Barrier 9.4 Combination of Models 8.1.2 In Vitro Excised Models 9.4.1 Primary Cell Cultures 168 168 172 176 182 183 183 184 187 187 192 201 203 204 205 216 217 218 218 219 219 219 220 220 221 221 222 223 223 224 224 224 229 235 236 236 239 240 240 .1.3 Factors Affecting the Nasal Absorption 9.3 Effect of Formulation 9.1 Transcellular Route 9.1 In Vivo Animal Models 9.1 Physiological Factors 9.2 In Vitro Studies 7.1 Anatomy of the Lung Airways 10.2 Drug Delivery to the Tracheo-Bronchial Region 10.2 Structure and Physiology of the Nasal Cavity 9. In Vitro Screening Models to Assess Intestinal Drug Absorption and Metabolism Sven Deferme.5.2 Buccal Epithelial Cell Cultures 7.4 In Vitro Primary and Passaged Cell Culture Models 9.2.6 Conclusions 10.3.1 Isolated Buccal Tissue Mounted in Diffusion Cell Apparatus 7.4.3.3 Ex Vivo Models (Ussing Chambers Technique) 8.2.1 Introduction 10.2 In Vitro Models to Study Intestinal Absorption 8.5.5.1 General Factors Influencing Intestinal Drug Absorption 8.5 Models for Nasal Drug Transport and Absorption Studies 9.1 Introduction 9. Ben Forbes.1 Anatomy and Function 9.2. In Vitro Cellular Models for Nasal Drug Absorption Studies Dae-Duk Kim 9.2.

and Sub-Conjunctival Ocular Drug Delivery 13. Inner Blood–Retinal Barrier: Transport Biology and Methodology Ken-ichi Hosoya and Masatoshi Tomi 14.4 Corneal Cell Culture Models 12.4.1 In Vitro Models of the Alveolar Epithelial Barrier 11.1.2 In Vitro Models of the Alveolar Epithelial Barrier for Drug Absorption Studies 11. and Kwang-Jin Kim 11.3. Kwang-Jin Kim.1 Introduction 12.2 Primary Culture Models of Conjunctival Epithelial Cell Layers 13. Michael Laue.1 Primary Corneal Cell Cultures 12. Lee 13.2 Anatomy and Physiology of Human Cornea 12.6 Concluding Remarks 13.2. In Vitro Models of the Alveolar Epithelial Barrier Carsten Ehrhardt. and Vincent H.4 Models for Studying Conjunctival Transport Properties 13. Gukasyan.4.3 Concluding Remarks 12.3 Conjunctival Disease Models 13.3 An Overview of Conjunctival Disorders 13. The Conjunctival Barrier in Ocular Drug Delivery Hovhannes J.1 The Alveolar Epithelium 11.1 Introduction 11.2 Trans.5 Organotypic Equivalents 12.1 Drug Absorption Studies 10.2 The Use of Alveolar Epithelial Cells in Biopharmaceutical Research 11.Contents ix 10.3.3 In Vitro Models of CF Airway Epithelium 10.2 Tracheo-Bronchial Epithelial Cell Lines 10.2.1 Excised Conjunctival Tissues 13.2 Immortalized Continuous Cell Lines for Corneal Epithelial Cells 12.4.3 Use of In Vitro Models of Tracheo-Bronchial Epithelial Barriers in Biopharmaceutical Research 10.1 Introduction 241 242 243 243 245 249 258 259 259 266 266 269 274 283 284 285 289 290 290 291 294 300 307 308 310 312 313 313 316 317 317 321 322 .2.2 Drug Metabolism Studies Using Tracheo-Bronchial Epithelial Cells 10.4 Concluding Remarks 11.5 Summary and Conclusions 14.1 Introduction to the Ocular Surface and the Relative Contribution of the Conjunctiva 13.L. Cell Culture Models of the Corneal Epithelium and Reconstructed Cornea Equivalents for In Vitro Drug Absorption Studies Stephan Reichl and Ulrich Becker 12.2.3 Transcorneal Drug Absorption into the Eye 12.4.4.

7 Conclusions 15.6 Regulation of the RLIS 15.3.5 Regulation of RTJ 15.6.2 Estrogen Regulation of Actin Polymerization 15.1 Introduction 15.x Contents 14.6.3 Regulation of Paracellular Transport Across Secretory Epithelia 15.1 Blood-to-Retina Influx Transport 14.3 Placental Transport Mechanisms 324 326 326 327 328 330 332 332 334 334 339 340 341 344 345 346 346 349 349 350 351 352 353 354 354 355 356 357 357 368 369 370 370 .6.5 Ex Vivo Transporter Gene Expression Levels at the Inner Blood–Retinal Barrier (Magnetic Isolation of Retinal Vascular Endothelial Cells) 14.2 Retinal Uptake Index Method 14.8 Concluding Remarks 16. Gorodeski 15.5.5 Composite Effects of Estrogen on Paracellular Permeability 15.7 Implications of the Data for Topical Drug Delivery 15.1 Role of Cao 15. Regulation of Paracellular Permeability in Low-Resistance Human Vaginal-Cervical Epithelia George I.1 Integration Plot Analysis 14.6.1 Introduction 16.4 In Vivo Vitreous/Retina-to-Blood Efflux Transport (Microdialysis Study) 14.6.6 Mechanism of Drug Transport at the Inner Blood–Retinal Barrier 14.3 Modulation of Actin Polymerization: Estrogen vis-à-vis Aging Effects 15.2 Structure of the Placenta 16.4 Early Stages of Tight Junctional Disassembly 15.2 Regulation of Assembled Tight Junctions by Extracellular ATP 15.5.5.1 Estrogen Regulation of the RLIS 15.2 Retina-to-Blood Efflux Transport 14. Audus 16. In Vitro Models and Multidrug Resistance Mechanisms of the Placental Barrier Pallabi Mitra and Kenneth L.6.3.5 Claudin/Occludin Model of Regulation of the RTJ 15.6.5.2 Conditionally Immortalized Cell Lines as a Novel in Vitro Inner Blood–Retinal Barrier Model (Uptake Studies) 14.4 Estrogen Regulation of Cortical Myosin 15.2 The Ussing-Zerahn Model of Transepithelial Fluid Transport 15.5.3 In Vivo Blood-to-Retina Influx Transport 14.3 Estrogen Regulation of RTJ 15.4 Regulation of Paracellular Permeability 15.

1 Measuring Drug Transport Across Epithelial Barriers in Submersed Conditions: Ussing Chamber and Transwell R -Like Systems 371 371 372 374 377 377 381 383 386 397 398 398 399 400 403 404 405 406 406 410 410 418 418 419 425 426 427 430 431 431 431 436 442 443 445 .2.2 Connecting Dissolution and Permeation Measurement in One Instrumented Setup 19. Michael Bur.2 Structure and Function of the Blood–Brain Barrier 17.5 Concluding Remarks 19. Motz.1 Perfused Placental Model 16.1 Introduction 18. Sam Wainhaus.3.2 Multidrug Resistance-Associated Proteins 16. and Kuo-Chi Cheng 18.4.3.2.3 Critical Evaluation of the State of the Art and Further Needs 19. High-Throughput Screening Using Caco-2 Cell and PAMPA Systems Cheng Li.1 Rapid Transport Protein Modulation 17.1 Introduction 17.1 Introduction 19.4.4 Available Model Systems 16. Annette S.3 Relevance of the Barrier for Drug Delivery 17.5.1 Discovery of the Blood–Brain Barrier 17.5.4.2 Brain Capillary Endothelial Cell Culture 17. Schaefer.4 Models to Study Blood–Brain Barrier Function 17.6 Conclusions 17. and Claus-Michael Lehr 19.3 Trophoblast Cultures 16.2 Modulation of P-glycoprotein Transcription 17. In Vitro Models to Study Blood–Brain Barrier Function Gert Fricker 17. Instrumented In Vitro Approaches to Assess Epithelial Permeability of Drugs from Pharmaceutical Formulations Stephan A. Ulrich F.5 Multidrug Resistant Transporters of the Placenta 16.3 Parallel Artificial Membrane Permeability Assays 18.3 Permeability Assessment of Pulmonary Aerosol Formulations 19.2 Caco-2 Cell Monolayer System 18.5. Uss.4.2.3.2 Intestinal Permeability of Drugs Delivered as Solid Dosage Forms 19.4.4 Combining Use of Caco-2 and PAMPA 18.4.5 Perspectives 18.1 Isolated Cerebral Capillaries 17.1 Rationale for Connecting Dissolution and Permeation Measurements 19.Contents xi 16.3 Breast Cancer Resistance Protein (BCRP/ABCG2) 16.3 In Silico Methods 17.1.1 MDR1/P-Glycoprotein (ABCB1) 16.2 Trophoblast Tissue Preparations 16.

2. Physiologically Based in Silico Models for the Prediction of Oral Drug Absorption Fredrik Johansson and Ronnie Paterson 21.3.1 Introduction 20.3 Critical Evaluation of the State of the Art and Further Needs Part 3 Bioinformatics—In Silico Tools to Predict Drug Absorption 20.2 Setups Allowing to Measure Drug Transport Across Pulmonary Epithelia Interfacing Air 19.3 Drug Absorption 21.2.2 Drug Dissolution 21.6 Outlook 21. Jitender Verma.1 Overall Absorption Process 21.4 The Use and Validation of Physiologically Based Simulation Models 21.1 Introduction 21.2 In Silico Methods Published for Prediction of BBB Permeability 22. In Silico Modeling for Blood–Brain Barrier Permeability Predictions Prabha Garg.3.1 How Predictive are the Models? 21.2 Absorption Process 21.3 Mass Balance Models 21.4.1 Introduction 22.3.2 The Skin Barrier 20.xii Contents 19.3 The Diffusion Equation 20.5 Pharmacokinetic Models 20.4.2 Mixing Tank Models 21.3.3.3.3 Physiologically-Based Absorption Models 21.4.4 Compartmental Absorption and Transit (CAT) Models 21.5 Other Models (PK-MapTM /PK-Sim R ) 21.1 General Considerations 21.1 QSPR Models 20.3.5 Summary 22.2 Situations Where Less Accurate Predictions Can Be Encountered 21.4. Modeling Transdermal Absorption Dirk Neumann 20.2 Non Steady-State Solutions and Morphological Models 20.3 Summary 446 450 459 460 460 461 461 463 477 478 479 486 487 488 488 489 490 491 491 492 494 496 499 500 500 502 505 510 511 512 544 .4 Data Analysis 20.2. and Nilanjan Roy 22.

4. Smart 25.1 PEPT (SLC15) 23.1 Exploiting Caveolae for Drug Delivery 25.Contents xiii Part 4 Molecular/Sub-Cellular Level—Mechanistic Tools 23.3.3 Caveolae and Vesicular Drug Transport 25.2 Determination and Classification of Drug Transporters 23.3. and Potocytosis 25.2 Cloning Techniques 24.6 Mammalian Cell Lines 24.3.5 Folate Conjugates and Therapeutics 25. Impact of Drug Transport Proteins Tomohiro Terada and Ken-ichi Inui 23.4 Caveolae.1 Introduction 24.4 Homology Cloning 24.1 Caveolin Expression 25.2 OATP (SLCO) 23. Everson.5 Insect Cell Lines 24.3. The Pharmacology of Caveolae Stuart A.5 Pharmacology at Caveolae: Presentation.2.1 Introduction 25.2 Caveolae-Linked Endocytosis and Non-Caveolae. and Eric J.3.2.2 A Caveolar Mechanism of Multidrug Resistance 25.3 Heterologous Expression Systems 24. Clathrin-Independent Endocytotsis Offer Delivery of Drugs to Novel Intracellular Targets 25.4 MATE (SLC47) 23. Receptor Clustering.2.3 Characteristics of Major Drug Transporters 23.1 Historical Overview and General Considerations 24.3 Yeast Expression Systems 24.4.4.5 ABC Transporters 23.3 A Gateway to Targeted Cancer Cell Ablation 25.2 Xenopus Laevis Oocytes for Cloning of Drug Carrier 24.3.4.3.4 Concluding Remarks 25.3.2 Bacterial Expression Systems 24.4.3 OCT/OCTN/OAT/URAT (SLC22) 23.2 Action at Caveolae 25. Ross.3. Presentation. Folate Receptor.3.1 Introduction 23.4 Xenopus Oocytes 24.2.3 Complementation Cloning Strategies 24.4 Caveolae and Cancer 25.3.4 Conclusions and Perspectives 24.3. William V. Cloning and Functional Heterologous Expression of Transporters Carsten Kneuer and Walther Honscha 24. Presentation 559 560 561 561 561 563 565 567 567 569 577 578 578 578 580 584 585 588 588 590 591 592 593 593 596 598 599 602 602 602 603 604 604 605 606 607 608 608 .1 Cell-Free Expression Systems 24.3.

Binding-Uptake Studies and Cellular Targeting Franz Gabor and Michael Wirth 27.1.3 Protocols 26. updated semi-annually.2.2. Esther H. Drug Permeability Studies in Regulatory Biowaiver Applications Donna A.3 Human Epithelial Cells as a Model for Transformation and Immortalization 26.2 Cell Transformation and Immortalization Strategies 26.4 Cytoevasion 27.4 Summary 27.5 Conclusions Appendix: Lab Protocols (available online.2. Volpe 28. as required) Index 616 617 618 618 618 619 619 621 621 626 640 641 642 643 643 645 645 647 648 650 650 651 652 652 655 659 665 666 667 669 672 676 681 683 .3.2 Selecting the Appropriate Cell Line 26.2 The Cell–Target System Interaction 27.6 Uptake and Intracellular Localization Within Acidic Compartments 27.1 Flow Cytometry 27.2.1 Design of Targeted Drug Delivery Systems 27. Isolation.2.5 Active or Passive Uptake? 27.4 Method Suitability 28.1 Generation.2.2.2 Intracellular Localization by Means of Confocal Laser Scanning Microscopy 27.1 Introduction 27. Immortalization Strategies for Epithelial Cells in Primary Culture David de Semir Frappart.2 Specificity of Interaction 27.2 Cell Culture Models 27.4 Concluding Remarks Part 5 Regulatory Considerations 28.1. Vock.3 Analytical Tools: Labeling and Detection of Targeted Drug Delivery Systems 27.2. and Dieter C. Gruenert 26.2.2 Regulatory Application of the BCS 28.1 Biopharmaceutics Classification System 28.2.3 Permeability Measurements 28.3 Binding Versus Uptake—Cytoinvasion Assays 27.1 Introduction 26.1 Transformation Versus Immortalization 26.xiv Contents 26.1.3. Rosalie Maurisse.1 Cytoadhesion Assays 27.3 Analytical Techniques 27. and Characterization 26.3.4 Strategies to Generate Transformed and/or Immortalized Epithelial Cells 26.

due to perhaps the central role played by such models in ‘translational research’. Andrea Macaluso from Springer Publishers contacted us with an invitation to edit a book on preclinical models for drug absorption and related subjects. Consuming unknown amounts of wine. followed by a dinner at an Italian trattoria in Sandymount. the FDA Biowaiver based on the Biopharmaceutics Classification System (BCS) has further boosted the use of in vitro absorption models. with a brief synopsis of pertinent data generated for each model. the brain. from epithelium to endothelium. if there is something that any reader might feel wrong about the present form(at) of the book. The book starts with chapters emphasising in situ and ex vivo techniques. the skin to various epithelial tracts of respiratory. we wanted to offer the reader a comprehensive overview of different models and approaches that can be utilised in the investigation and prediction of drug absorption via many biological barriers in the body. Moreover. the outline for the book was scribbled on several pieces of paper serviettes provided by the trattoria’s patient staff. Later that year. which now appear to enjoy a renaissance in research. This book is meant to provide an overview on what is available with regards to various experimental models for the studies of drug transport in general. when KwangJin Kim visited Trinity College Dublin where Carsten Ehrhardt teaches and pursues his research. In vitro absorption models have revolutionised our understanding in this field and pushed our thinking more in terms of mechanisms beyond mere phenomenology. We have tried to cover as many cell-based in vitro approaches as possible.Preface It all started on a day in summer of 2005. we strongly blame it on the Italian wine which we drank during the brainstorming session part of the working dinner. further evolving into the wish to predict the drug’s fate at a preclinical stage during drug development.e. as some questions cannot be answered with certainty using simplistic in vitro models. that is in silico models that have been established for several barriers in the very recent past. we were able to come up with three chapters on cell-free systems. gastrointestinal and reproductive organs. that is relevant topics and potential authors. when Ms. for those originating from almost all known biological barriers of importance (i. Hence. By the end of the dinner. Over the recent years. the eye and so forth). In short. Moreover. So as to give the reader who may be keen on utilising New Age Science tools sans the biological tissues and cells altogether. we also included a section that xv . we went out for drinks after a hard day’s work. we enjoyed rather yummy Irish-Italian dishes and worked on the basic layout of the book. the impact of absorptive/secretive processes on a drug’s bioavailability has been recognised.

. this book also features a ‘living appendix’. since the information on these techniques might be useful for the more advanced or more molecular biology-oriented users. Finally. transporter proteins or endocytotic processes). We the editors send our warmest regards to all the authors who had to put up with our constant nagging and repeated demands on several rounds of revisions. an idea born out of most likely too much vino flown down into our digestive tracts during the brainstorming session at the trattoria.. it may have caused some anxiety attacks. We would like to make a note that a chapter focusing on regulatory considerations of using the absorption models and FDA Biowaiver implications is also included. Therefore. To some authors. but not the least. As a final twist. binding and uptake. in that the reader can print a respective protocol that she/he wants to use in the daily run of laboratory works. both Carsten Ehrhardt and Kwang-Jin Kim appreciate the expert review and digestion for the in silico chapters extended by their mutual friend. thus. as the science moves forward. we can hardly wait for the next edition for that reason.D. albeit on a voluntary basis as need arises. The actual format and question and answer sessions may evolve as time goes by. Ian Haworth. alleviating the fear of the physical/chemical damage incurred on the expensive book at the laboratory bench side.xvi Preface focuses on several techniques for the studies of drug transport mechanisms (i. for which we apologise and also commend them profusely for their patience and hard work. and the generation of continuous cell lines. It is our intention to urge the participating authors to make necessary updates/additions to their respective appendix materials. Hence. Ph. Last. we also realise that the book is a live being that constantly requires updates and mending.e. we expect that the appendices that are going to be available by the time the book is in print will always reflect the latest trends/techniques. at the University of Southern California – School of Pharmacy. We might also establish an online forum where readers/users can post pertinent questions or request specific protocol(s).

Ph. Dr. E-mail: sven. Pieter Annaert. Laboratorium voor Farmacotechnologie en Biofarmacie. K-15-2-2700. USA. School of Pharmacy. Ph..D. Lawrence. Fax: +1 908 740 2916. Telephone: +32 16 330 303.deferme@pharmaxl.augustijns@pharm. Fax: +49 681 302 4677. Laboratorium voor Farmacotechnologie en Biofarmacie. Dipl.D. 3000 Leuven. Germany.becker@labtec-pharma.D. E-mail: pieter.ca Prof.ac.com Cynthia Bosquillon. Katholieke Universiteit Leuven. Department of Pharmaceutical Chemistry.agu@dal. 3000 Leuven.bus 921.net xvii . Nova Scotia B3H 3J5. KS 66045. Strijderstraat 7. The University of Kansas. College of Pharmacy. Gasthuisberg .V. Telephone: +32 16 39 47 96. 5968 College Street. University Park.D. Germany.bosquillon@nottingham.bur@mx. Biopharmazie und Pharmazeutische Technologie.edu Prof. NJ 07033. Telephone: +1 908 740 4056. Agu. 2056 Malott.. Telephone: +32 16 330 301.D.. Ulrich Becker.be Dr. Telephone: +44 115 8466078. Patrick Augustijns.. 2015 Galloping Hill Road. Telephone: +1 902 494 2092. Dalhousie University.-C. USA. Universität des Saarlandes. Schering-Plough Research Institute. Fax: +32 16 330 305. E-mail: cynthia. Telephone: +49 681 302 3140. Fax: +1 902 494 1396. Belgium. Raiffeisenstr. Boots Science Building Room D10. Belgium.cheng@spcorp. Belgium. Telephone: +1 785 864 3591. Katholieke Universiteit Leuven. E-mail: kuo-chi. Audus. Ph.kuleuven. Halifax. 1251 Wescoe Hall Drive. University of Nottingham. Cheng.annaert@pharm. nat. E-mail: ulrich.. 4a. O&N2. E-mail: m. Ph. Gasthuisberg . Fax: +1 785 864 5265. Departement Farmaceutische Wetenschappen.bus 921. Gebäude A4 1. rer. Labtec GmbH.de K. Pharm. Fax: +32 16 330 305. O&N2. Nottingham NG7 2RD. Kenilworth. Pharma XL Comm. E-mail: audus@ku. Canada. E-mail: patrick. Dr. 66123 Saarbrücken.Contributors Remigius U. Ph.uk Dr. 3370 Boutersem.be Kenneth L. nat. E-mail: remigius. Fax: +32 16 39 47 01. Telephone: +49 2173 9735 25.uni-saarland. Fax: +49 2173 9735 35.com Sven Deferme. Im Stadtwald... Burbidge Pharmacy Building. School of Pharmacy.kuleuven. 40764 Langenfeld. Departement Farmaceutische Wetenschappen. Michael Bur. rer.

CA 92121. Victoria 3052. Franklin Williams Building. nat. University College Cork. University MacDonald Women’s Hospital. Lexington. Telephone: +43 1 4277 554 06. California Pacific Medical Center Research Institute. Departments of Reproductive Biology. E-mail: brendan. Institut für Pharmazie und Molekulare Biotechnologie. rer.griffin@ucc.A. Fax: +44 207 848 4800. USA. Panoz Institute.. Ruprecht-Karls. Telephone: +1 859 323-2852. Fax: +49 6221 54 5971. College Road. University Hospitals of Cleveland.. Telephone: +44 207 848 4823. E-mail: barrie. Ph. Telephone: 216-844-5977.. Germany.S. Trinity College Dublin. 423 Sanders-Brown. Carsten Ehrhardt. Ph. Cavanagh Pharmacy Building. S. Cork. Telephone: +353 1 896 2441. E-mail: prabhagarg@niper.de Univ.in Prof. San Francisco. Fax: 216983-0091.D. nat. Suite 220. E-mail: franz. Fax: +353 896 2783. Franz Gabor. KY 40536. Ph.edu. E-mail: hovik. Ph. Department für Pharmazeutische Technologie und Biopharmazie. Finnin.D. E-mail: dieter@cpmcri.Universität Heidelberg. Fax: +61 3 9903 9583.. Universität Wien. E-mail: ben. USA. USA.gukasyan@pfizer. Gruenert. Telephone: +91 172 2214682-87. Ireland. Department of Pharmaceutics. Everson. Fax: +91 172 2214692. Dr.. Department of Pediatrics.fricker@uni-hd.ie William V. Ohio 44106. UK. Telephone: +1 858 622 5998.E.. School of Pharmacy and Pharmaceutical Sciences. Ph. Ireland.ac.D.forbes@kcl. King’s College London. Abteilung für Pharmazeutische Technologie und Pharmakologie.Tech.. Fax: +1 415 600 1725.xviii Contributors Dr. University of Kentucky.D.. Austria.-Prof. Gert Fricker.uk Prof.ac. Cleveland. 69120 Heidelberg. Gukasyan.D.au Ben Forbes. Room 224. 381 Royal Parade.finnin@vcp.org Hovhannes J. Gorodeski. Pharmaceutical Science Division. Im Neuenheimer Feld 366. Nagar. Physiology and Biophysics.D. Parkville.ie Dieter C. M. Fax: +43 1 4277 9 554.edu Brendan Griffin. La Jolla Laboratories.com . Telephone: +49 6221 54 8336. Telephone: +61 3 9903 9520.D.. Science & Technology. George I. and the Department of Obstetrics and Gynecology. M. 150 Stamford Street.. E-mail: gig@cwru. National Institute of Pharmaceutical Education & Research. Pfizer Global R&D. B. 1090 Vienna. E-mail: gert.edu Barrie C. CA 94107. Case Western Reserve University (CWRU). Computer Centre. Ph. Ph.D. Research Formulations. Ph. Monash University. Fax: +1 859 257 2120.ac. Sector 67. E-mail: ehrhardc@tcd. Fax: +353 21 490 1656.gabor@univie. Pfizer Inc.at Prabha Garg. Fax: +1 858 678 8185. Dr. 800 Limestone Street.D. Dublin 2. rer.. NIPER..monash. Punjab 160 062. San Diego. 11100 Euclid Avenue. Telephone: +1 415 600 1362. Westland Row. Victorian College of Pharmacy. Althanstrasse 14. Telephone: +353 21 490 1657. Pharmaziezentrum. Australia. London SE1 9NH. 475 Brannan Street. India. and Oncology. E-mail: everson@uky. School of Pharmacy.

Ph. Seoul 151-742. Chinese University of Hong Kong. School of Pharmacy. USA. 221 87 Lund. Campus.D. Japan. Telephone: +49 341 9738 132. University of Southern California.uni-leipzig. 04103 Leipzig. South Korea. Telephone: +81 76 434 7505. Fax: +49 341 9738 149. 13353 Berlin. E-mail: ddkim@snu. Institut für Pharmakologie.u-toyama.fjo.L. Fax: +49 1888 754 2571. Fax: +82 2 873 9177. Im Stadtwald. Department of Pharmacy. 2011 Zonal Avenue. Hong Kong. Ph. Pharmaceutical and Analytical R&D. Michael Laue. E-mail: honscha@vetmed. 66123 Saarbrücken. Telephone: +49 1888 754 2326. Shatin. Seoul National University. Fax: +46 46 33 7128.hansen@mx.D. E-mail: hosoyak@pha.edu. Biopharmazie und Pharmazeutische Technologie.. Universität des Saarlandes. Dr. Pharmazie und Toxikologie. PA 19341. Faculty of Medicine. Hidalgo. Germany. Will Rogers Institute Pulmonary Research Center. nat. Exton. Telephone: +49 681 302 3140. E-mail: fredrik.kyoto-u. E-mail: inui@kuhp. Shogoin. Institut für Pharmakologie. Dr.uni-saarland. Oaklands Corporate Center. Ph. Telephone: +82 2 880 7870.jp Fredrik Johansson. Gebäude A4 1.J. Universität des Saarlandes. Toyama 930-0194.de .. rer. Telephone: +1 323 442 1217. 440 Creamery Way. E-mail: Kwang. Ph.ac. College of Pharmacy. Dipl. 66123 Saarbrücken. USA. Germany. Germany. E-mail: vincent.lee@cuhk.Kim@usc. Biopharmazie und Pharmazeutische Technologie. Telephone: +852 2609 6862. Robert Koch-Institut.D.. 2630 Sugitani.Johansson@astrazeneca.D. Fax: +1 610 280 3779.ac.kr Kwang-Jin Kim.de Ismael J.ac. Sweden. Ph.uni-saarland. Los Angeles.de Vincent H. Japan. Kyoto University Hospital. Germany. Nordufer 20. E-mail: kneuer@vetmed. Walther Honscha. Pharm. E-mail: hidalgo@absorption.edu Dr. nat.D. Germany. Zentrum für Biologische Sicherheit 4 (ZBS4). An den Tierkliniken 15. Fax: +81 76 434 5172. Fax: +1 323 442 2611. Room 626E.. Kyoto 606-8507. nat.Contributors xix Steffi Hansen. E-mail: st. Suite 300. Claus-Michael Lehr. Gebäude A4 1.de Dr. Graduate School of Medical and Pharmaceutical Sciences. Fax: +852 2603 5295. Telephone: +46 46 33 7671. An den Tierkliniken 15. AstraZeneca R&D Lund. Lee. CA 90033. Telephone: +81 75 751 3577. Sakyo-ku.com Dae-Duk Kim. Ph. 54 Kawahara-cho. rer. Keck School of Medicine. Pharmazie und Toxikologie. Telephone: +49 341 9738 142. Universität Leipig. Fax: +49 341 9738 149. Department of Pharmaceutics. nat. Fax: +49 681 302 4677.de Ken-ichi Hosoya..D. E-mail: lauem@rki. Absorption Systems.. 04103 Leipzig.hk Prof.com Prof.. New Territories. University of Toyama.D.uni-leipzig. Ph. Universität Leipig. 6/F Choh-Ming Li Basic Medical Science Building. E-mail: lehr@mx. Room HMR-914. Fax: +49 681 302 4677.jp Ken-ichi Inui.. rer. Fax: +81 75 751 4207. Telephone: +1 610 280 7300. Carsten Kneuer. Telephone: +49 681 302 3039. rer.

Kenilworth. E-mail: ejsmart@email. Suite 300. KY 40536. E-mail: caitriona. Institut für Pharmazeutische Technologie. Ph. Telephone: +353 91 70 1128. PA 19341.reichl@tu-braunschweig. nat. School of Pharmacy. Email: tanja_obradovic@merck.com Dr. Ross. Ireland. Absorption Systems. rer. Telephone: +1 610 280 1406. 66123 Saarbrücken.D.de Stuart A.. K-15-2-2700. Dirk Neumann.nicolazzo@vcp.Sc. USA. 351 North Sumneytown Pike. KG.maurisse@gmail. USA. Fax: +61 3 9903 9583. Telephone: +49 6805 9292 402. nat. Telephone: +1 908 740 4363. Telephone: +1 415 600 1362.. 226 Simons Laboratories. R&D Galway. Fax: +1 415 600 1725. Universidad de Concepción. 440 Creamery Way. Ph.D. University College Cork. Lawrence. 66129 Saarbrücken.com Pallabi Mitra. Ph.edu Dr. Ph. Departamento de Farmacia. Mendelssohnstraße 1..au Caitriona O’Driscoll. Cork. Fax: +353 21 490 1656. Lexington. The University of Kansas. Zentrum für Bioinformatik. Telephone: +61 3 9903 9605. Department of Pediatrics. Schering-Plough Research Institute. Parkville. Technische Universität Carolo-Wilhelmina zu Braunschweig.. NJ 07033. Inc. Telephone: +1 785 864 4138. North Wales. rer. KS 66047. Telephone: +353 21 490 1396.xx Contributors Cheng Li.cl Rosalie Maurisse. E-mail: joseph. UG4D-80.paterson@bsci. Ireland. Motz. E-mail: rosalie. nat..monash. Phone: +1 267 305 1576. Ph.de Joseph A. Oaklands Corporate Center. Department of Pharmaceutical Chemistry. E-mail: ronnie. 423 Sanders-Brown. Gebäude E1 1. Ph. PA 19454-2505. E-mail: li. Fax: +49 531 391 8108. Fax: +1 908 740 2916.edu.com Dr.com) Ronnie Paterson. Ursapharm GmbH & Co.D. Stephan Reichl. Ballybritt Business Park. University of Kentucky. Fax: +1 859 257 2120. M. Telephone: +1 859 323 6412. E-mail: pmitra77@ku. Germany. Fax: +1 215-305-6428.uky. School of Pharmacy. USA. Chile. Australia. Industriestraße. Concepción.uni-sb. Fax: +49 6805 9292 88.. Galway. USA. 800 Limestone Street. Fax: +49 681 302 64719.D. Germany. Fax: +1 610 280 3779. 2015 Galloping Hill Road.D.de Dr. Suite 220. CA 94107.. Barrio Universitario s/n.neumann@bioinf. Stephan A. Exton.D.motz@ursapharm.edu .. Victoria 3052. Fax: +56 41 2207086.odriscoll@ucc. rer.. 475 Brannan Street. Universität des Saarlandes.. Victorian College of Pharmacy. Monash University. Javiana Luengo Contreras.ie Tanja Obradovic. College Road.Merck & Co.cheng@spcorp. E-mail: d.com (current contact information . Im Stadtwald. nat. Telephone: +49 531 391 5651. Boston Scientific Ireland.. Germany. E-mail: obradovic@absorption. 38106 Braunschweig.Sc. Telephone: +56 41 2203019.D. E-mail: s. Fax: +353 91 70 1395. Fax: +1 785 864 5736. Ph. Nicolazzo. rer. E-mail: s. Department of Pharmaceutics. Cavanagh Pharmacy Building. Telephone: +49 681 302 68610. 2095 Constant Avenue. California Pacific Medical Center Research Institute. M. E-mail: jluengo@udec. 381 Royal Parade. USA. San Francisco.

Clinical Development. E-mail: ugwoke@disphar. NIPER. Telephone: +49 7351 54 97912.. KG. Department of Dermatology.S. Sweden. 423 Sanders-Brown. E-mail: annette. Department of Pediatrics. (Pharm.D.co.. Uss. Fax: +91 172 2214692.kyoto-u. NJ 07033. Punjab 160 062. E-mail: esther. Sector 67. Sector 67.. Campus. E-mail: ann. Telephone: +49 681 302 2019. Telephone: +81 75 751 3586.uni-saarland.D.D. Smart. M. CA 94115. Telephone: +91 986 7589850. San Francisco.schneider@mx. University of Kentucky.S. Schering-Plough Research Institute. 54 Kawahara-cho. E-mail: jeetu270@yahoo. Japan. Kenilworth.A.uky. 2340 Sutter Street. Sakyo-ku. S-221 87 Lund. Dr. Ph.D. Vock.Sc.edu Tomohiro Terada.in Dr. Biopharmazie und Pharmazeutische Technologie. Fax: +1 908 740 2916. S. Nagar. Marc Schneider. Telephone: +1 908 740 3696. National Institute of Pharmaceutical Education & Research. Ph. 3741 LM Baarn. nat. K-15-2-2700. Telephone: +91 172 2214682-87. USA... San Francisco.de Prof. Shogoin. Fax: +81 75 751 4207. Ph. 66123 Saarbrücken.com Annette S. Ugwoke. E-mail: tomimasatoshi@yahoo.uss@spcorp. University of Toyama. Tel: +31 35 5280 429.in Dr. E-mail: tomohiro@kuhp.Contributors xxi Nilanjan Roy. nat. Fax: +46 46 33 6666.jp Masatoshi Tomi. Universität des Saarlandes.ac. E-mail: ejsmart@email. Gebäude A4 1. Department of Pharmaceutics. Ph.com Michael I. Graduate School of Medical and Pharmaceutical Sciences. Kyoto University Hospital. Germany. Fax: +1 859 257 2120.com Jitender Verma. Nagar. Punjab 160 062. Fax: +81 76 434 5172. USA.ucsf. Birkendorfer Straße 88397 Biberach an der Riß. The Netherlands. E-mail: ddesemir@cc. Japan. AstraZeneca R&D Lund. Telephone: +46 46 33 6482. 800 Limestone Street. Germany. National Institute of Pharmaceutical Education & Research.com .D. 2015 Galloping Hill Road. UCSF Cancer Center..co. Boehringer Ingelheim Pharma GmbH & Co.tronde@astrazeneca. E-mail: nilanjanroy@niper. Campus.boehringeringelheim.ac. Schaefer. nat.de David de Semir Frappart. Pharmacoinformatics Division. Esther H.D. Pharmazeutische Nanotechnologie. India. Fax: +91 172 2214692. Universität des Saarlandes. Ulrich F. Telephone: +81 76 434 7508. E-mail: ufs@rz. Tolweg 15. Germany. Lexington. rer. E-mail: marc. rer. Fax: +1 415 502 8218. Toyama 930-0194. Ph. Telephone: +1 415 502 5638. Kyoto 606-8507.jp Ann Tronde.. Disphar International bv.). Ph. Ph. Suite 471. USA. Gebäude A4 1. Fax: +49 681 302 4677. S. Telephone: +49 681 302 2438. University of California. Fax: +49 681 302 4677. Telephone: +1 859 323 6412. India. sc. 66123 Saarbrücken.D.uni-saarland. NIPER. Ph. Department of Biotechnology. Fax: +31 35 5480 585. 2630 Sugitani. KY 40536. Non-Clinical Drug Safety.A. Department of Pharmacy.vock@bc.edu Eric J.D.

Department of Biopharmaceutics. Food and Drug Administration. USA. MD 20993. E-mail: donna. NJ 07033. Althanstrasse 14. Silver Spring. Mizuho-ku. E-mail: yuasa@phar.com Research Institute. Schering-Plough 15-2-2700. Fax: +1 301 796 9816. Österreich. Ph. +1 908 740 2916. KKenilworth. Telephone: +43 1 4277 554 07.gov Sam Wainhaus. Graduate School of Pharmaceutical Sciences.jp . 10903 New Hampshire Avenue.D. E-mail: michael. Telephone: +81 52 836 3423. Fax: samuel. Ph.wainhaus@spcorp.D. 2015 Galloping Hill Road.volpe@fda. USA.ac.nagoya-cu. Volpe. 3-1 Tanabe-dori. Fax: +81 52 836 3426.. Department für Pharmazeutische Technologie und Biopharmazie. Telephone: +1 908 740 6587.D. Dr.wirth@univie.hhs. Michael Wirth. Ph.ac.-Prof.. 1090 Wien. E-mail: Univ. Fax: +43 1 4277 9 554.xxii Contributors Donna A. Telephone: +1 301 796 0014.. Pharmaziezentrum. Universität Wien. Nagoya 467-8603. Nagoya City University. Japan.at Hiroaki Yuasa.