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Percutaneous

Penetration Enhancers
Drug Penetration
Into/Through the Skin

Methodology and
General Considerations

Nina Dragicevic
Howard I. Maibach
Editors

123

Percutaneous Penetration Enhancers
Drug Penetration Into/Through the Skin

Nina Dragicevic  •  Howard I. Maibach
Editors

Percutaneous
Penetration Enhancers
Drug Penetration
Into/Through the Skin
Methodology and General
Considerations

Editors
Nina Dragicevic Howard I. Maibach
Apoteka "Beograd" San Francisco
Belgrade California
Serbia USA

ISBN 978-3-662-53268-3    ISBN 978-3-662-53270-6 (eBook)
DOI 10.1007/978-3-662-53270-6

Library of Congress Control Number: 2017937381

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Preface

The main function of skin is the protection of the body from the external
environment by preventing loss of water and the ingress of exogenous sub-
stances. This implies that the skin acts as a barrier for the diffusion of sub-
stances into the underlying tissue. Despite this role, the skin has become
recognized as an important drug delivery route which can be reached directly.
It is an ideal site for the application of drugs for achieving local (topical) and
systemic (transdermal) drug effects. Local or topical drug delivery assumes
treating various skin diseases, while transdermal delivery aims to achieve
systemically active drug levels in order to treat systemic diseases. Drugs have
been applied to the skin to achieve also regional drug delivery which involves
drug application to the skin to treat or alleviate disease symptoms in deep tis-
sues beneath the skin (such as in musculature, etc.). Topical and transdermal
drug delivery offer a number of advantages compared to other conventional
routes, and hence they are of great interest to pharmaceutical research, which
explains the increasing interest in skin as a site of drug application.
However, skin represents a formidable barrier for percutaneous drug
absorption, being of crucial importance for achieving topical and transdermal
effects of drugs. Significant efforts have been devoted to developing strate-
gies to overcome the impermeability of intact human skin. There are many
ways for circumventing the stratum corneum, which provides the main bar-
rier to drug penetration. These methods can be divided into chemical and
physical penetration enhancement methods, i.e., percutaneous penetration
enhancers(as well as the skin structure are described in the other volumes in
this book series).
The aim of this book is to provide to readers working in academia and
industry, including young researchers, an up-to-date comprehensive work
describing all the important topics required to understand the principles of
enhancing transdermal and dermal drug delivery. We have divided this book
into five volumes.
Volume 1 begins with a description of the skin, as understanding of its
structure, function, and especially its penetration pathways is fundamental to
understanding how topical and transdermal dosage forms work and how dif-
ferent methods may be employed and work to enhance percutaneous drug
penetration as well as how they work. The first parts devoted to skin and the
stratum corneum, representing its uppermost layer being responsible for its
protection, discuss their structure, the importance of the lipid organization in
the stratum corneum, the different penetration pathways through the skin

v

vi Preface

with a focus on the increasing importance of the follicular route, as well as
the influence of different excipients on the skin. The first volume of this book
focuses on the chemical methods used to overcome the impermeability of
intact skin, such as different drug manipulation strategies (drug or prodrug
selection, chemical potential control, eutectic systems, complexes with cyclo-
dextrines, etc.) and influences of formulation/vehicle effects (influences of
emulsions, nanoemulsions, Pickering emulsions, microemulsions, emulsifi-
ers, emollients, liquid crystalline structures, gels, etc.) on the penetration
enhancement of drugs.
Volume 2 describes, similarly to Volume 1, the chemical methods used in
penetration enhancement of drugs. However, this volume is devoted to the
application of different kinds of nanocarriers and represents an attempt to
familiarize the readers with the importance of nanocarriers used to enhance
the percutaneous penetration of drugs as they have numerous advantages in
comparison to conventional drug formulations. More recently, different types
of nanocarriers have been designed by researchers which allow controlled
and targeted drug delivery (dermal or transdermal drug delivery), improved
therapeutic effectiveness, and reduced side effects of drugs. As carriers they
can be classified into lipid-based vesicles (e.g., liposomes, transfersomes,
invasomes, etc.), surfactant-based vesicles (e.g., niosomes, novasomes, and
others), lipid-based particulate carriers (e.g., solid lipid nanoparticles, nano-
structured lipid carriers, and lipid nanocapsules), polymer-based particulate
carriers (e.g., polymeric nano- and microparticles, polymeric nanocapsules,
polymeric micelles, dendrimers, dendritic core-multishell nanocarriers, etc.),
nanocrystals, and others. This volume therefore focuses on the different
nanocarriers and gives a comprehensive review of their use as promising drug
delivery systems. It also considers the use of nanocarriers for cutaneous
immunization offering the important advantage of being painless and having
a stronger immune response compared to the intramuscular injection of vac-
cines. In addition, the volume provides insights on the safety of the topical
use of nanoparticles.
Volume 3, similarly to Volumes 1 and 2, describes the chemical methods
used in penetration enhancement of drugs with an emphasis on the enhancing
methods used to modify the stratum corneum. It starts with the classification
of penetration enhancers and their mode of action and provides insights on
the structure–activity relationship of chemical penetration enhancers. The
focus of this volume is on the most commonly used classes of skin penetra-
tion enhancers being investigated in scientific literature and used in commer-
cial topical and transdermal formulations, and their representatives are
discussed in more detail, including their mechanism of action, where known.
The following penetration enhancers are considered in this volume: alcohols
(e.g., ethanol), glycols (e.g., propylene glycol), amides
(1-­dodecylazacycloheptan-2-one or laurocapram (Azone®)), fatty acids (e.g.,
oleic acid, etc.), fatty acid esters (e.g., isopropyl myristate, etc.), ether alco-
hols (e.g., diethylene glycol monoethyl ether (Transcutol®)), pyrrolidones
(N-methyl-2-pyrrolidone), sulphoxides (e.g., dimethyl sulphoxide, etc.), sur-
factants (e.g., polysorbates, etc.), terpenes (e.g., L-menthol, etc.), peptides

Preface vii

and new classes of enhancers, such as iminosulfuranes, transcarbams,
­dimethylamino acid esters, and dicarboxylic acid esters. In addition, syner-
gistic effects of different chemical penetration enhancers have been discussed
in this book as an important feature of chemical penetration enhancers.
Furthermore, the safety profile of chemical penetration enhancers is
considered.
Volume 4 considers the current status and possible future directions in the
emerging area of physical methods being used as potent enhancers for the
percutaneous penetration of drugs. It gives a comprehensive overview of the
most used methods of enhancing dermal and transdermal drug delivery. It
covers sonophoresis, iontophoresis, electroporation, magnetophoresis,
microneedles, needle-free jet injectors, ablation methods (electrical, thermal,
or laser skin ablation), and others. The numerous advantages of these meth-
ods have opened new frontiers in the penetration enhancement of drugs for
dermal and transdermal drug delivery. Cutaneous vaccination and gene deliv-
ery by physical methods have been also discussed in this volume.
Consideration was given to new methods, too, such as a novel electrochemi-
cal device for penetration enhancement, different waves (e.g., photoacoustic
waves, microwaves, etc.), natural submicron injectors, moxibustion, and oth-
ers. Furthermore, the combined use of different physical methods or of physi-
cal methods and passive enhancement methods (chemical penetration
enhancement methods) are discussed as they provide, due to their synergistic
effects, higher percutaneous drug penetration when used together.
Volume 5 provides fundamental principles of the drug penetration into/
through the skin, from covering basic mathematics involved in skin perme-
ation of drugs, influences of drug application conditions and other factors on
drug penetration, mechanistic studies of penetration enhancers, influences of
the type of skin used (human native or reconstructed skin) to different meth-
ods utilized to assess the drug penetration into/through the skin and to deter-
mine the amount of permeated drug (such as tape-stripping of the stratum
corneum, electron spin resonance, Raman spectroscopy, attenuated total
reflection, confocal laser scanning microscopy, single and multiphoton
microscopy). Retardation strategies are also discussed as being important for
some classes of drugs, such as sunscreens. The safety of applied penetration
enhancers as well as the research ethics in the investigation of dermal and
transdermal drug delivery are addressed in this volume. This volume dis-
cusses the current status and future perspectives of passive/chemical and
active/physical penetration enhancement methods as they are gaining exten-
sive interest as promising tools to enable an efficient dermal or transdermal
drug delivery.
We are very thankful to all authors who contributed chapters to the book
series Percutaneous Penetration Enhancers, as they found time to work on
the chapters despite having busy schedules and commitments. All the authors
are eminent experts in the scientific field which was the subject of their chap-
ter, and hence their contribution raised the value of this book. We also sin-
cerely thank our publishing, developmental, and production editors Portia
F. Wong, Ellen Blasig, Sverre Klemp, Andre Tournois, Grant Weston, and

viii Preface

others from Springer, for their dedicated work which was necessary to achieve
such a high standard of publication. We highly appreciate readers’ comments,
suggestions, and criticisms to improve the next edition of this book.

Belgrade, Serbia Nina Dragicevic
San Francisco, CA, USA Howard I. Maibach

Riviere 7 Mechanistic Studies of Permeation Enhancers ��������������������������  119 S. El Maghraby 3 Finite and Infinite Dosing��������������������������������������������������������������   35 Wing Man Lau and Keng Wooi Ng 4 Non-formulation Parameters That Affect Penetrant-Skin- Vehicle Interactions and Percutaneous Absorption��������������������   45 Jeffrey E. Ulrich F. Elizabeth Ryan. Mechanisms. Claus-Michael Lehr and Steffi Hansen 2 Occlusive Versus Nonocclusive Application in  Transdermal Drug Delivery����������������������������������������������������������   27 Gamal M. Hamid R. and Michael S. Isha Haridass. Moghimi. Yousuf Mohammed. Higuchi 8 High Throughput Screening of Transdermal Penetration Enhancers: Opportunities. and Samir Mitragotri ix . Qian Zhang. Chittenden and Jim E. Grice. Roberts 6 The Effects of Vehicle Mixtures on Transdermal Absorption: Thermodynamics. Jeffrey E. Methods. Leite-Silva. Kevin Li and William I. Roberts 5 The Influence of Emollients on Dermal and Transdermal Drug Delivery ��������������������������������������������������������������������������������   77 V. Schaefer.R. Grice. Pankaj Karande. Yousuf Mohammed. Hamid R. and Michael S. Moghimi.Contents Part I  Factors Influencing Percutaneous Drug Penetration  1 Basic Mathematics in Skin Absorption����������������������������������������    3 Dominik Selzer. and Prediction����������������������������������������������������������  95 Jason T. and Applications����������������������������������������������������������������������������  137 Amit Jain. Assessment.

and Claus-Michael Lehr 11 Stripping Procedures for Penetration Measurements of Topically Applied Substances ��������������������������������������������������  205 Jürgen Lademann. and Yves Chevalier 14 ATR-FTIR Spectroscopy and the Skin Barrier: Evaluation of Penetration-Enhancement Effects������������������������  247 Julia Covi-Schwarz. Marie-Alexandrine Bolzinger.x Contents Part II  Methods for Measuring the Percutaneous Drug Penetration  9 Models. Storage Capacity of Nanotransporters. Prow 17 Corneoxenometry: A Bioassay Exploring Skin Barrier Breaching��������������������������������������������������������������������������  303 Claudine Piérard-Franchimont. and Alfred Fahr 16 Clinical Cutaneous Drug Delivery Assessment Using Single and Multiphoton Microscopy ��������������������������������  283 Anthony P. Schaefer. Sabine Schanzer. S. and Drug Release��������������������������������������  215 Stefan F. Haag. Jürgen Lademann. and Gérald E. Piérard Part III  The Retardation of Percutaneous Drug Penetration  18 Retardation Strategies for Sunscreen Agents������������������������������  311 Katharina Bohnenblust-Woertz and Christian Surber 19 Retardation of Dermal Release by Film Forming Emulsions������������������������������������������������������������������������  321 Dominique Jasmin Lunter and Rolf Daniels . Heike Richter. D. Selzer. Meinke 13 Confocal Raman Spectroscopy as a Tool to Investigate the Action of Penetration Enhancers Inside the Skin����������������  229 Stéphanie Briançon. Daya D. Methods. Ashtikar. Hansen. Hans-­Jürgen Weigmann. and Measurements in Transdermal Drug Delivery����������������������������������������������������������  153 Donald M. Meinke. and Alexa Patzelt 12 Application of EPR-spin Probes to Evaluate Penetration Efficiency. Victoria Klang. and Martina C. Raphael and Tarl W. Trinh Hermanns-Lê. Verma. Martina C. and Claudia Valenta 15 Confocal Microscopy for Visualization of Skin Penetration������������������������������������������������������������������������  255 Mukul A. Cropek and Pankaj Karande 10 Human Native and Reconstructed Skin Preparations for In Vitro Penetration and Permeation Studies ����������������������  185 Ulrich F.

Navin C. Sinduja C. Donnelly Part V Safety Issues and Ethics in Studies on Dermal and Transdermal Delivery 23 Efficacy. Hamid R. Moghimi.Contents xi Part IV  Current Status of Dermal and Transdermal Drug Delivery  20 Penetration-Enhancement Strategies for Dermal and Transdermal Drug Delivery: An Overview of Recent Research Studies and Patents��������������������������������������  337 Syed Sarim Imam and Mohammed Aqil 21 Perspectives on Dermal Delivery of Macromolecular Drugs ����������������������������������������������������������������������������������������������  355 Marianna Foldvari and P. Chandrasekaran. Safety and Targets in Topical and Transdermal Active and Excipient Delivery��������������������������������  369 Yousuf H. Grice. Mohammed. Roberts 24 Ethical Considerations in Research Involving Dermal and Transdermal Drug Delivery��������������������������������������������������  393 Dusanka Krajnovic and Nina Dragicevic Index��������������������������������������������������������������������������������������������������������  405 . and Michael S. Wedad Sakran. Kumar 22 Active Enhancement Methods in Transdermal Drug Delivery: Current Status and Future Perspectives ��������������������  359 Ryan F. Shereen A. Sukumar. Césa R. Jeffrey E. Bibi. Yousef.

Switzerland Marie-Alexandrine Bolzinger  University of Lyon. Egerkingen. University of Queensland. Ashtikar  Lehrstuhl für Pharmazeutische Technologie. Germany Césa R. Cary. IL. Université Claude Bernard Lyon 1. Laboratoire d’Automatique et de Génie des Procédés (LAGEP). Northern Ireland. UK Dusanka Krajnovic  Department for Social Pharmacy and Pharmaceutical Legislation. Hamdard University. Université Claude Bernard Lyon 1. France Jason T. NC. Chittenden  Department of Biomathematics.Contributors Mohammed Aqil Department of Pharmaceutics. NC. Bibi  Therapeutics Research Centre. School of Medicine. USA Rolf Daniels Department of Pharmaceutical Technology. India Mukul A. CNRS UMR 5007. Construction Engineering Research Laboratory. Champaign. Medical Biology Centre. Army Corps of Engineers. New Delhi. Villeurbanne.S. Australia Katharina Bohnenblust-Woertz  Galderma Spirig. Cropek  Environmental Chemistry Laboratory. North Carolina State University. Woolloongabba. Australia Yves Chevalier  University of Lyon. USA Donald M. Queens University Belfast. Eberhard Karls University. CNRS UMR 5007. QLD. Woolloongabba. School of Medicine. Villeurbanne. France Stéphanie Briançon  University of Lyon. Raleigh. Université Claude Bernard Lyon 1. Donnelly  School of Pharmacy. Princess Alexandra Hospital. Belgrade. USA Product Development. Engineering Research and Development Center. Faculty of Pharmacy. France Navin C. CNRS UMR 5007. Princess Alexandra Hospital. Germany Ryan F. QLD. Tuebingen. University of Queensland. Friedrich-­ Schiller-­Universität Jena. U. Jena. Belfast. Chandrasekaran Therapeutics Research Centre. Pharsight Corporation. University of Belgrade Faculty of Pharmacy. Laboratoire d’Automatique et de Génie des Procédés (LAGEP). Laboratoire d’Automatique et de Génie des Procédés (LAGEP). Serbia xiii . Villeurbanne.

Center of Experimental and Applied Cutaneous Physiology. School of Medicine. Belgium William I. USA Pankaj Karande  Howard Isermann Department of Chemical and Biological Engineering. India Amit Jain  Department of Chemical Engineering. Saarbrücken. Germany Isha Haridass Therapeutics Research Centre. Center of Experimental and Applied Cutaneous Physiology. School of Pharmacy. Woolloongabba. Waterloo. USA Victoria Klang University of Vienna. Liège University. Serbia Gamal M. Troy. University of Waterloo. QLD. Canada Jürgen Lademann Charité – Universitätsmedizin Berlin. Woolloongabba. CA. UT. Kumar  Helix BioPharma Inc. Jena. Saarland University.xiv Contributors University of Belgrade – Center for the study of Bioethics. SK. Venereology and Allergy. The Glocal University. Department of Dermatology. Princess Alexandra Hospital. University of Queensland. Princess Alexandra Hospital. Saskatoon. Technology and Biopharmaceutics. Rensselaer Polytechnic Institute. Germany Wing Man Lau  School of Pharmacy. Higuchi  Pharmaceutics and Pharmaceutical Chemistry. Salt Lake City. UK Claus-Michal Lehr Biopharmaceutics and Pharmaceutical Technology. University of Utah. Department of Pharm. Austria P. Berlin. Venereology and Allergy. Foldvari  School of Pharmacy. NY. College of Pharmacy.. Egypt Alfred Fahr  Lehrstuhl für Pharmazeutische Technologie. University of Tanta. Helmholtz Institute for Pharmaceutical Research Saarland. Reading. Tanta. Santa Barbara. USA Syed Sarim Imam  Department of Pharmaceutics. Friedrich-Schiller-­ Universität Jena. Department of Drug Delivery. School of Medicine. University of Reading. Germany Steffi Hansen  Biopharmaceutics and Pharmaceutical Technology. Saarbrücken. Saarland University. Belgrade. Saarbrücken. Canada Jeffrey E. Australia Trinh Hermanns-Lê Laboratory of Skin Bioengineering and Imaging. Haag Department of Dermatology. Berlin. QLD. Germany . Germany M. University of California – Santa Barbara. El Maghraby Department of Pharmaceutical Technology. Saharanpur. Liège. Uttar Pradesh. Charité – Universitätsmedizin Berlin. Vienna. University of Queensland. College of Pharmacy. Australia Stefan F. Center for Biotechnology and Interdisciplinary Sciences. Grice Therapeutics Research Centre. ON.

Princess Alexandra Hospital. School of Medicine. USA .R. USA Hamid R. Kevin Li Division of Pharmaceutical Sciences. Brisbane. Brisbane. University of Reading. Liège University. University of California – Santa Barbara. Reading. Diadema. Leite-Silva  Universidade Federal de São Paulo. Piérard  Laboratory of Skin Bioengineering and Imaging. Translational Research Institute. Australia School of Pharmacy. Germany Gérald E. School of Medicine. Australia Keng Wooi Ng  School of Pharmacy. USA Dominique Jasmin Lunter Department of Pharmaceutical Technology. Germany Martina C. Meinke  Charité – Universitätsmedizin Berlin. Woolloongabba. Venereology and Allergy. Prow Dermatology Research Centre. Germany Samir Mitragotri Department of Chemical Engineering. Kansas State University. Liège. Princess Alexandra Hospital. QLD. Germany V. Berlin. Center of Experimental and Applied Cutaneous Physiology. Instituto de Ciências Ambientais Químicas e Farmacêuticas. Department of Dermatology. Shahid Beheshti University of Medical Sciences. Translational Research Institute. Department of Dermatology. Saarbrücken. Tehran. Raphael  Dermatology Research Centre. OH. Belgium Claudine Piérard-Franchimont Laboratory of Skin Bioengineering and Imaging. Venereology and Allergy. Moghimi Therapeutics Research Centre. Center of Experimental and Applied Cutaneous Physiology. Princess Alexandra Hospital. Riviere  Institute of Computational Comparative Medicine. CA. Brazil S. Berlin. UK Alexa Patzelt Charité – Universitätsmedizin Berlin. Germany Jim E. Berlin. Iran Yousuf H. The University of Queensland. Eberhard Karls University. Venereology and Allergy. KS. QLD. Cincinnati. Manhattan. Santa Barbara. School of Pharmacy and Medical Sciences. University of Queensland. Liège University. QLD. Australia Anthony P. University of South Australia. School of Medicine. SA. SP.Contributors xv Department of Drug Delivery. Belgium Tarl W. Department of Anatomy and Physiology. College of Pharmacy. Tuebingen. The University of Queensland. Center of Experimental and Applied Cutaneous Physiology. Australia Heike Richter Charité – Universitätsmedizin Berlin. University of Cincinnati. College of Veterinary Medicine. Adelaide. Department of Dermatology. Helmholtz Institute for Pharmaceutical Research Saarland. Liège. Mohammed  Therapeutics Research Centre.

Princess Alexandra Hospital. Woolloongabba. Schwarz  University of Vienna. Australia Christian Surber  University Hospital Zürich. Australia Qian Zhang Therapeutics Research Centre. Vienna. Helwan.. Center of Experimental and Applied Cutaneous Physiology. Adelaide. Zürich. Saarbrücken. Research Platform ‘Characterisation of Drug Delivery Systems on Skin and Investigation of Involved Mechanisms’. Princess Alexandra Hospital. Verma  Novartis Pharmaceutical Corp. Venereology and Allergy. Technology and Biopharmaceutics. Australia Therapeutics Research Centre. University of Queensland. Germany Sabine Schanzer Charité – Universitätsmedizin Berlin. School of Medicine. Vienna. Department of Dermatology. Department of Dermatology. QLD. QLD. Germany Sinduja C. NJ. Princess Alexandra Hospital. Princess Alexandra Hospital. Research Platform ‘Characterisation of Drug Delivery Systems on Skin and Investigation of Involved Mechanisms’. School of Medicine. QLD. East Hanover. University of Queensland. University of Queensland. University of South Australia. University of South Australia. Helwan University. SA. Berlin. Department of Dermatology. Schaefer Biopharmaceutics and Pharmaceutical Technology. Center of Experimental and Applied Cutaneous Physiology. Roberts Therapeutics Research Centre. Germany Shereen A. School of Pharmacy and Medical Sciences. Saarland University. Egypt Ulrich F. School of Pharmacy and Medical Sciences. School of Medicine. USA Hans-Jürgen Weigmann  Charité – Universitätsmedizin Berlin. Sukumar  Therapeutics Research Centre. Switzerland University Hospital Basel. Woolloongabba. Austria University of Vienna. Saarbrücken.xvi Contributors Michael S. Australia . Venereology and Allergy. Woolloongabba. Adelaide. Berlin. Woolloongabba. Saarland University. SA. School of Medicine. Basel. Austria Daya D. Department of Dermatology. Yousef Therapeutics Research Centre. Germany Julia C. Austria Dominik Selzer  Biopharmaceutics and Pharmaceutical Technology. Australia Elizabeth Ryan Therapeutics Research Centre. Australia Wedad Sakran  School of Pharmacy. QLD. Switzerland Claudia Valenta  University of Vienna. Department of Pharm. University of Queensland. Vienna.

Part I Factors Influencing Percutaneous Drug Penetration .

.. 10 css kg/m3 Steady-state concentration 1.............6... Maibach (eds............5 In Vitro–In Vivo Correlation (IVIVC).........-M....................... 12 Initial concentration at t = 0 1...2. coefficient Saarland University......2. Germany kp m/s Permeability coefficient e-mail: d..2 Numerical Diffusion Models.. 17 J Diffusion flux mol 1....1 Infinite Sums in Analytical Solutions...........................de © Springer-Verlag Berlin Heidelberg 2017 3 N......1007/978-3-662-53270-6_1 .... 16 formulation 1.. 21 1................ H........3 Analysis of Skin Penetration.. 6 AUC kg/m3 Area under the curve 1.. Saarbrücken... solute in the membrane st..uni-saarland..... ufs@mx..uni-saarland..2 Analysis of Skin Permeation............... Ulrich F..............de kt % applied Transfer coefficient C......4 Advanced Mathematical Approaches D m2/s Diffusion coefficient for Studying Skin Absorption.......................... and Steffi Hansen Contents Table of Abbreviations 1..hansen@mx...de.. Selzer • U... l m (Macroscopic) thickness Saarland University....4........... 21 m2 s Conclusion...........2 Dealing with Finite Dose Skin Permeation. 12 C0 kg/m3 1........ Schaefer Ko/w ................ Germany Mss kg Steady-state amount of e-mail: lehr@mx............. Germany m kg Mass Department of Drug Delivery....selzer@mx. 21 Jmax mol Maximum diffusion flux 1. DOI 10.F..... Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin.1 Skin-Concentration Depth Profiles........1 Laplace Domain Solutions.... 22 m2 s k J/K Boltzmann’s constant K ..1 Dealing with Infinite Dose Skin Permeation....... Schaefer.3............. Lehr • S........2 Fitting of Experimental Data........ Hansen (*) s Biopharmaceutics and Pharmaceutical Technology.3.....uni-saarland.... Dragicevic.)..................uni-saarland...... Claus-­Michael Lehr... 8 c kg/m3 Concentration 1.....2 Skin Compartmental Approaches.......6 Tips and Tricks........ 22 Jpeak mol Peak flux References.... 16 hv M Height of applied 1........ Octanol–water partition Biopharmaceutics and Pharmaceutical Technology..de.. Helmholtz Institute M kg Mass per area for Pharmaceutical Research Saarland... 4 A m2 Area of application 1..I....6... Basic Mathematics in Skin Absorption 1 Dominik Selzer... Saarbrücken... Saarbrücken..4... 19 m2 s 1..1 Mathematical Background of Analyzing Skin Absorption Symbol Generic units Description Processes.......................... Partition coefficient D..... 15 Cl m3/h Systemic clearance 1..........

1 Mathematical Background cal models to describe transdermal drug of Analyzing Skin transport. corneocyte desquamation (Reddy et al.5 that cally described by laws derived by Adolf Fick allow the application to a wider range of in 1855. (Eq.2): kT dx .3. tions with certain initial conditions).4 and c the concentration at point x in space and (Crank 1975). tial to sites of lower chemical potential ((Eq. the diffusion coefficient. Solutions of these parabolic partial Absorption Processes differential equations can be solved analyti- cally for various initial and boundary condi- It is usually assumed that the travel of mole. D dj ( x.1) problem. S s Saturation concentration t s Time 1987).4) derive Fick’s second law of diffusion (Eq. c ( x. Inhomogenous media transitions change the clas- sical diffusion problem to a diffusion-partition J ( x. diffusion is based on Brownian motion setup (e. binding phenomena (Frasch r m Radius et al.2) simpli- η Pa s Viscosity fies to Eq..t ) (1. for example.t ) d 2 c ( x . Selzer et al. 1.4 D.2) M¥ ¶t MW kg Molecular weight In this general equation. the diffusion coefficient ODE . enzymatic reactions (Guy et al.t ) = . time t. is a t is directed from sites of higher chemical poten- proportionality constant usually given in m2/s. ble but complex models in Sect. 1.t ) kg Applied mass = Ñ ( D Ñ c ( x . one can J ( x. Pharmacokinetic address. Here.t ) =D (1. This is a From an empirical and more macroscopic fundamental issue and must be always kept in understanding.g. b).t ) Assuming conservation of mass. 1. Additional terms can be included to PK .t ) ) (1. 1. in mol/m2 s) to the concentration case. Symbol Generic units Description ¶c ( x. the diffusion flux at a cross section x at time gradient.t ) (1.3) mol K dt d x2 Homogeneity is often a strong simplifica- tion but a typical assumption for easy analyti- 1. From an atomistic point the description of a specific experimental of view. Fick’s first law (Eq. We will shortly discuss more flexi- diffusant in a medium and can be mathemati. Anissimov and Roberts 2004)).. tions and are presented in Sect. 2012).g. Ordinary differential may vary in dependence of location and/or con- equation centration. As cules through the skin membrane is governed mentioned earlier. 1.D Ñc ( x. D. or general convective transport to tpeak s Time to peak flux model elimination or clearance of molecules into T K Temperature the systematic circulation (Dancik et al. these conditions often simply by passive diffusion due to the absence denote simplifications and are only true for of active transporters. infinite dose or finite dose condi- of particles in virtue of their thermal energy. scenarios.t ) = . V m3 Volume For the one-dimensional case and a homogenous x m Space medium (constant diffusivity). 1. tlag s Lag time 2000a. the driving force of molecular mind when applying mathematical models in movement is a concentration gradient of the general.3 φ J Chemical potential d c ( x. 2011). 1.1) relates the diffusion For the inhomogeneous and more general flux J (e.

6). but if the at least a basic understanding of the relationship solubilities in the two phases are relatively low between molecular properties and the aforemen. 1. 1.7) (Eq. t) is the chemical potential of the sub. 1.t ) = kT ln ( c ( x. MW is the solute molecular weight. K  dx  is a thermodynamic parameter reflecting the Besides studies with variable partition and/or relative affinity of a solute for a certain phase diffusion coefficients inside a certain skin layer over another phase. φ(x. 1.n (1.3.with the help of a linear free-energy relationship Dm = Dm0 MW . 1. and evaluation of transder. ing diffusivities in the skin domain. Where the diffusivity D denotes the speed and the position-­ dependent partition coefficient of a diffusant through a membrane and is K is given by Eq. it is more ficient between layer m2 and layer m1 can be ­common (especially for more complex ­multilayer defined as Eq. and k is Boltzman’s and Dm0 are constants. and n stance.5 (Anissimov and Roberts inversely related to the weight of the solute. it could be found parameter of interest. the 2004): partition coefficient K accounts for jumps in concentrations at the interface of two adjacent  d  d c ( x. the characteristics of the constant. In this chapter. 1.t )  skin layers and is often related to a measure of =   solute lipophilicity. In general. 1.7) (Anderson and could be related to the stratum corneum lipid Raykar 1989): phase/water partition coefficient Klip. the interested reader is kindly referred to diffusion-partition problem with the chemical ­ Hansen et al.t ) / K ( x ) . For deeper The general case equation that describes the insight. the partition coef- (Anissimov and Roberts 2004). ries derived from polymer research have also Eq. A prominent example is empirically that for a small particle the following the ocanol/water partition coefficient Ko/w that relationship holds (Eq. 2013).4 simplifies to the standard diffusion equa. Therefore. fitted. Hence.8) ­coefficients to the interfaces of two adjacent lay.10) (Anderson et al. with Eq. Here.9 the diffusion coefficient can be calculated by S m2 using the well-known Stokes–Einstein relation K m2 / m1 = (1.t d ln K x  (1. setup. the partition diffusivities and partition coefficients. this effect can tioned parameters is essential when it comes to be neglected. S m1 and S m2 in the different layers (Anissimov For spherical particles in a continuous fluid.t ) = kT ln(c ( x. ( ) potential defined as j ( x.1  Basic Mathematics in Skin Absorption 5 Here. with viscosity of the solvent η and par. 1988) . T is the temperature. or experimentally determined phases at equilibrium.t ) and a constant diffusivity. (which holds true for many cases). (2013). and the partition coefficient can be the preparation. been applied successfully to the field of describ- tion as stated in Eq. For the homogeneous case with membrane at a specific temperature. Cmeq1 ers.6) between an easy-to-observe parameter that 6ph r mimics the membrane properties and the For diffusion in polymers.8 models) to assign specific diffusion coefficients Cmeq2 to the different layers and specific partition K m2 / m1 = (1. coefficient is concentration-dependent.t ) d  D ( x ) d x c ( x. the presented solutions of the diffusion equation and determination of distinc. 1.5)  ( ) ( ) ( ) rithmic octanol–water partition coefficient. S m1 ticle radius r: A typical strategy is to establish a relationship kT D= (1.9) (Eq. et al. with Cmeq2 and Cmeq1 being the respective solute tive parameters almost exclusively depend on concentrations at the interface of the two adjacent predicted. Other theo- j ( x. defined by using the saturation concentrations mal skin transport. most commonly the loga- dt d x  − D x c x.

conditions of the acceptor compartment are ments. semi-infinite (vol- erning mechanics. profiles for the infinite. neered skin or artificial skin surrogates (for a erties of the layers at the interface (e.1 An aqueous donor/acceptor Permeation experiments are usually performed to with a drug diffusion coefficient of measure the amount permeated through the bar. donor compartment.10). 2 of SC or viable epidermis/stratum corneum). and parti- an in vitro setup. The initial donor concentration was set to important measure. like the well-known vertical Franz DSkin® http://www. formulation ( £ 10 μl/cm2 of a liquid formulation) such as Eqs. K lip = aK ob/ w (1. again refer the interested reader to Hansen When it comes to application scenarios. These theo- 1.10) diffusion cell (Franz 1975) and horizontal Bronaugh cell (Bronaugh and Stewart 1985) as Here. see characteristics of the relationship between Sect. ume > 10  μl/cm2. lipid path length of 180 μ m was assumed. one et al. respectively for an area of diffu- treatment of tissue beneath the site of sion of 1. which Permeation experiments are typically applied corresponds to a swollen membrane for an using diffusion cells yielding in a general separa.6 D. For diffusion coefficient of 1. finite dose experiments are and applying mathematical models as well as defined by an application of a limited volume of when trying to generalize basic relationships. This is an tions. Perfect sink tion of the diffusion domain in different compart. namely donor..767 cm2. underlying physics.com 1  .g. routes through the stratum corneum or implicit 2002). Typical barrier mem- aqueous vehicle and lipid phase of the stratum branes for investigation consist of excised human corneum.1. mathematically eral transport mechanisms (e.. compartments. donor/ detailed overview. Selzer et al. 6. we kindly refer to Sect. In the case of infinite dosing. according to the binding phenomena) of the diffusant.7) and (1.g. Therefore. and the donor volume for the semi- regional drug delivery through the skin (e.3 of Chapter 10).33E-5 cm2/s.scientific-consilience. but with a depletion of the donor which is already perceptive). Since literally all (OECD 2004a. Organisation for Economic Cooperation and tant fact must be kept in mind when developing Development (OECD). 3.2 Analysis of Skin Permeation retical calculations were performed using the DSkin® software. Frequently used diffusion cells are static cells. For the finite dose case. 2001). We Chap. (1. b). parameters derived from Typical concentration/mass/flux versus time mathematical concepts might obfuscate the gov. 1.56 was used for simula- mass in an acceptor compartment. this relates to the accumulated tion coefficient of 6. the constants a and b correspond to the well as flow through cells (for further details. A tortious stratum corneum application). the applied dose is assumed to be so large that It must be noted that representation of diffu. evaporation or diffusion through the barrier does sivity and partition coefficient in certain skin only negligibly change the concentration in the ­layers by a single number basically averages sev. especially for systemic and 1 mg/ml. these values range from 1 to 10 mg/cm2. several different the dose is assumed to be infinite (Brain et al. in vitro setup (Talreja et al. bioengi- strongly depends on the physicochemical prop. It is obvious that this relationship or animal skin in its various fashions. barrier. stratum corneum lipid channel rier over time in relation to the diffusion area. 16). For semisolid and solid formu- mathematical models heavily oversimplify the lations.. and finite dose cases are depicted in Fig.87E-8 cm2/s. for infinite dose and finite dose scenario was set as achieving therapeutic drug levels systemically or 2 μl and 20 μl.g. This impor. (2013) for an extensive overview about can distinguish between infinite dose and finite the determination of diffusion model input dose experiments. parameters. and acceptor assumed.

1. Obviously. In contrast. change of mass inside the (dashed line). This shows opposed to this. 1. 1. and finite dosing (dotted line).1a shows the eled through the membrane. 1. the mass of the finite dose case clearly that the assumption of an infinite dose (no decreases after reaching a m ­ aximum (Fig. This is mass inside the acceptor compartment reaches the crucial when applying the mathematical concepts typical straight ­steady-­state line (Fig. semi-infinite compartment over time (a).1  Theoretical change of concentration in the donor o­ utgoing flux (d) for infinite (solid line). were performed using the DSkin® software tor compartment (c). The barrier mass-over-­time curve for dose case shown in Fig. the applied volume an infinite dose setup does reach a plateau as per area was set slightly higher than the finite soon as the steady state is reached (Fig. This ­presented in the next subsections. 1. corresponds to a plateau of the flux-over-time pound of approximately 300 Da with a log KO/W profile (Fig. the flux characteristic depletion of the donor for the finite reaches a maximum and subsequently decreases dose case and less pronounced for the semi-infi. As dose threshold defined by the OECD. Simulations stratum corneum (b). The concentration-­ acceptor compartment if all substance has trav- over-time profile depicted in Fig.1b). and change of stratum corneum The chosen values correspond to a model com. For the simulation of the semi-infinite nite case. since c­ hoosing . the accumulated by applying fixed volume-based rules.1  Basic Mathematics in Skin Absorption 7 a b Donor Stratum corneum c d Acceptor Outgoing flux Fig. 1.1b).1d).1c). 1.1d. accumulated mass inside the accep. the finite dose sce- of 2 in an aqueous vehicle. significant depletion) does not automatically hold In case of the infinite dose setup. with time. and model parameters nario will reach a theoretical mass plateau in the were estimated by DSkin.

In the next two Fig. the absorption curve can be described ment over time. 1.1 D  ealing with Infinite Dose centration in the donor compartment. with rier. D l è 6D ø is the macroscopic diffusion coefficient. It has to be kept in mind that even if small volumes of highly concentrated solutions are applied and the per- Steady-state meation is low. part of the steady state (Fig. Crank (Crank 1975). 1. the solution simplifies to the linear the partition coefficient between donor and bar. and that no compound of interest is Analysis of infinite dose in vitro skin permeation located inside the barrier at time t = 0 . K is steadystate).12) significantly during experimental time periods.2. The first parameter is the . and t is the From Eq.2  Accumlated mass over time in the acceptor com- partment of an infinite dose experiment (solid line) and sections. 1. analytical solutions of the diffu- sion equation are always tailored to certain boundary and initial conditions). the system might behave like an Mass infinite dose case due to the fact that significant donor depletion only occurs at very long experi- mental time periods in relation to the permeated solute amount in the acceptor compartment. It is obvious that with t parameters when it comes to analysis of infinite leaning toward infinity (and hence reaching the dose experiments. Selzer et al. In this case. the reader is kindly referred to (see Fig. we assume a constant and steady con- 1. For axis denotes the lag time an exhaustive compilation of solutions regarding the diffusion equation for various boundary and initial conditions. A denotes the area of application.12.2).g. ÷ (1.2 êë 6 p å n2 exp ç - l 2 ø úû ÷ú (1. Investigation of infinite and finite dose experi- ments typically differ concerning their parame- ters of interest and application of mathematical Time concepts (e.. a few assumptions must be made to derive by J. analytical solutions of the diffusion linear part of the steady-state phase (dashed line). we can examine important time after application. with line (the steady state) with constant flux JSS é ( -1) æ Dn 2p 2 t ö ù n ¥ 1 2 m ( t ) = A ´ K ´ l ´ C0 ê K ´ l ´ t .11) n =1 è Here.2). perfect sink Skin Permeation conditions (zero concentration in the acceptor at all times). For short times. membrane. From a mathematical point of the excellent book The Mathematics of Diffusion view. C0 is the concentration of applied formula- DK æ l2 ö tion in the donor which is assumed not to change m (t ) = A C0 ç t . is typically done by measuring the cumulative By incorporating these rules for a homogeneous amount of substance inside the acceptor compart. Crank 1975). the amount by an analytical solution of Fick’s second law of increases exponentially until reaching a steady diffusion (Scheuplein 1967. an analytical solution for the diffusion equation by defining initial and boundary conditions. l is the macroscopic thickness of the barrier.. 1. faulty assumptions can lead to serious misinter- pretation of experimental data.8 D. The equation and mathematical concepts for the most intersection of the linearized steady-state phase and time typical experimental settings are presented.

of permeability coefficient and lag time can gen- cal vehicles. The permeability coefficient the skin and vehicle.7 (Crank property of a solute to permeate across a specific showed that steady state is achieved when medium (e.g. Hence. however.14) A further parameter to characterize infinite C0 dose absorption is the so-called lag time tlag given by Such a parameter might heavily depend on exper- l2 imental conditions. A word of caution is nec- sometimes misleading when comparing the per. Anissimov et al. making it an ideal parameter to com- defined as pare permeability of different solutes. The apparent per. by removing insufficient sink conditions (Anissimov and the influence of the partition coefficient between Roberts 1999). plished graphically by manual interpolation of bility (Eq. J SS kp = (1. under infinite dose and perfect sink conditions. 1. Jmax should be independent can be determined by dividing the slope of the of the vehicle applied. the skin) which is independent of Dt = 0. intrinsic permeability of a solute in a certain which is often given in units of cm/h and is medium. Obviously.13) affect the transport kinetics in the barrier (Zhang l et al. 1. determination between compounds which are applied in identi. it can be establish a steady state. ­optimally correlated to the molecular weight ter for the strength of permeation for a compound and lipophilicity of a compound (Fiserova- through a certain barrier from a specific vehicle Bergerova et al. essary concerning the meaning of tlag. 2013). introduced a erally be accomplished in two fashions: change of paradigm when it comes to explaining experimental data (Sloan et al. 1.15). This can be accom- solute through a barrier for comparing permea.15) important to mention that the occurrence of l physically untenable negative lag times often Here. Potts and Guy 1992. It is Sm J max = kp ´ S v = D ´ (1. Although kp may be a It is a measure that relates to the time it takes for useful and popular parameter when it comes to a compound to travel through the barrier and examination of permeation experiments.. l2 Therefore. 2011). and hence a direct parame. the dose but influenced by the applied vehicle. it can be meability coefficient kp describes an intrinsic approximated by multiplying tlag with 2.45 approximately (Crank 1975)). 2008). tlag does not meation of several compounds (Michaels et al. as long as the vehicle does not DK kp = (1. comparisons are only possible From a practical point of view. Sloan et al.2). McKone and Howd 1992). it describes an linear part by the initial concentration C0 and . it denotes a normalization of (2004) could show that molecular weight is the steady-state flux JSS with main determinant when it comes to predicting solute maximum flux.1  Basic Mathematics in Skin Absorption 9 s­o-­ called apparent permeability coefficient. Magnusson et al. The easiest approach utilizes the linearization gested to use the more expressive parameter Jmax of the steady state (see Fig. it is possible to the last data points that contribute to the steady overcome the limitations addressed before state or mathematically more soundly by a lin- ear fit (Schäfer-Korting et al. It is independent of the area of application and In contrast to epidermal kp which is initial concentration.16) mind when comparing parameters (interlabora. this is the case. kp. 1990. They sug. 2006). 1. Sv is the saturated permeant concentration indicates experimental problems such as in the vehicle and Sm is the solubility of the solute donor depletion (Barbero and Frasch 2009) or within the barrier. this has to be kept in tlag = (1. 6D tory and intralaboratory).2) which inter- which denotes the maximum possible flux of a sects the time axis at tlag. By using Jmax. Mathematically. Thus. In 2006. is achieved (see Fig. In other words. directly represent the time when the steady state 1975.

 1.2. l2 tions can overcome this problem. Often.21) As opposed to the infinite dose case. reach a steady state. Based 1. 1. application area A or simply by using the two discarding data often yields wrong results steady-state data points with and can produce a high variability. Henning et al. a Eq.2 å exp ( . ation experiments and showed the superiority lems arise using this approach that makes it of this procedure over the manual approach extremely prone to errors – from an experi. Kubota et al.18) and stepwise addition of data points from tend (last data point) toward t0 (first data D (1.17) tion.20) êë 6 p n =1 n 2 úû This reduces the number of unknown parameters. the swollen skin is not exactly known. Carelessly et al. 1. 2009). In 2009. 1993) é ( -1) ù n ¥ 1 2 m ( t ) = A × P1 × C0 × ê P2 × t .2 D  ealing with Finite Dose Skin on the theory of heat flow by Carslaw and Permeation Jaeger. a description of the flux of the com- pound leaving the barrier at time t is given by In contrast to the infinite dose exposure scenario. The clear advantage is the arise in data evaluation of infinite dose perme- simplicity in calculations. A huge advantage is the mental and evaluation point of view: ability to deliver sound results even if only a (a) This approach cannot be applied if the partial representation of the steady state is steady state was not clearly reached at the available.10 D. a frequent sampling P1 = K × l (1. Selzer et al.19) P2 = point) and analyzing the linear interpola. 1. inside the acceptor compartment at late time points mental data using a nonlinear least squares (see Fig.P2 n 2p 2 t ) ú (1.23) è .1a) and increase in flux until reaching the so-called peak flux Jpeak at time tpeak (Fig..2 ÷. per- the steepest part of the curve for evaluation formed an in-depth analysis of problems that (Buist et al. depend- ing on which points are chosen for evalua- mt1 . 1991. l ø (1. 1.11). kp = P1 × P2 (1.22) tion and application of an analytical solution tlag = 6 P2 for the description of absorption curves require a more sound mathematical foundation. By (b) It is often difficult to identify which data defining partition parameter P1 and diffusion points contribute to the steady state and parameter P2 with which do not. Often.mt2 kp = (1. (Henning et al. This is an obvious but often neglected AC0 ( t1 . (c) Using only a limited amount of data points and reformulating Eq. 1. A more sophisticated approach is the use of If the slope of the curve decreases after reaching the mathematical representation of the entire steady state. P1 and P2 can be easily fitted to experi. Permeability and lag time can be a characteristic depletion of donor concentration subsequently computed by (Fig. deduc- 1 (1.3). but builds a mass plateau Hence. 2.t2 ) problem in processing experimental data.1c). the macroscopic thickness of end of the experiment.11 yields (Díez-Sales generally ignores information. 2010). but several prob. a general strategy is to employ curve (Eq. Finite dose experiments also show approach. Cooper and typical finite dose absorption profile does not Berner 1985): D ¥ a i2 æ a i2 Dt ö J (t ) = 2 × A × M ¥ × b × å l 2 i =i cos a i ( b + b 2 + a i2 ) exp ç .23 (Carslaw and Jaeger 1959.

g.18 and case. To reduce the is finding the root of the first derivative of J(t) and number of unknowns.2b å exp ç. given by a i tan a i = b (1. Newton’s A general strategy to easily find Jpeak and tpeak method) (Kasting 2001) is key.29 simplifies to the familiar (Kasting 2001.2 ÷ (1.hv2 ) (1. and macro- ­regression to tabulated values of the first roots scopic path length l. The maximum of the curve denotes the peak Integrating Eq. A basic strategy to solve data by a nonlinear least squares approach and the transcendal equation is to use logistic yields values for β.24) hv Flux and the roots of the transcendal equation.. since Eq. 1. 1. diffusivity D. (1.29) equation to experimental data. Scheuplein and Ross 1974): expression: D l2 J peak = 1.85M ¥ .26) M¥ i =i cos a i ( b + b 2 + a i2 ) è l ø In comparison to Eq. reasonably small in comparison to the macro- For small doses ( b < ~ 0. Brent’s method (Brent 1973). If the thickness of the applied formulation is lem is.19) can be applied. 1. 1. the same approach that hence solving the following equation for tpeak: D ¥ a i2 æ a i2 Dtpeak ö a i2 D 2× M¥ × b × å l 2 i =i cos a i ( b + b 2 + a i2 ) exp çç - l2 ÷÷ 2 = 0 (1.1  Basic Mathematics in Skin Absorption 11 with l b =K .1 ).30) l2 6D In contrast to fitting solutions of the diffusion tpeak = (l 2 .26 can be fitted to experimental arbitrary values of i. (see Fig.25 must be solved for Equation  1.28) tpeak = (1.23 yields the accumulated flux Jmax at time to peak flux tmax mass per area (Kasting 2001) with M (t ) ¥ 1 æ a i2 Dt ö = 1 . The remaining parameters keep their Fig.root n .3  Sketch of outgoing flux across the barrier– meaning as introduced above. and hv is Time the height of the formulation in the donor com- partment. be calculated by the following simple equations and Eq. e. 1.26 requires a more skillful 1. some researchers 6D use the steepest linear part of the absorption curve .2 ) followed by a Jpeak and time to peak flux tpeak. αi.11 for the infinite dose worked for the infinite dose case (Eqs. 1.27) è ø l A reliable solution for the root-finding prob. To find further roots a subsequent Important parameters for evaluation of finite linear ­ extrapolation from previous roots dose permeation experiments are the peak flux ( root n = 2 ´ root n -1 . (1. 1. acceptor interface over time of a finite dose experiment (solid line).25) Here.4). Jpeak and tpeak can scopic diffusion path length. solving Eq. evaluation. refinement step (root-finding with. for example. M ¥ denotes the applied mass. it can be neglected. 1.

These reached (Fig. after a certain time by multiplying kt by t. 2000.3. 1. 2011). Wilkinson et al.12 D. a fitting of the right kinetic scenarios. since the flux changes with clearly differ for different exposure scenarios. (2012). 1. pseudo steady state with a straight line that will It has to be kept in mind that finite dose kinetics drop due to the depletion of the donor over time are also prone to variability of drug distribution in (Fig.11. a straight . Lademann For the finite dose case. One of the most prominent experimental tech- and the time to the steepest part to determine peak niques to obtain skin-concentration depth profiles flux and time to peak flux (van de Sandt et al. centage of dose per time (Eq. as for et al. in 1974. is tape-stripping (Stinchcomb et al. and refer- imprecisions of the manual method. and drug distribution to the nonincubated lateral parts in a Franz diffusion-cell setup (Selzer et al. Besides The infinite dose curves show the typical the fact that this parameter depends on the applied exponential decay for short exposure times and dose. 3. 1. 2008.5c). 2006). this a fast and relatively noninvasive technique to approach is prone to errors. Especially for long obtain absorption data for the in vitro and in vivo sampling intervals. Tape-stripping is the manual method for the infinite dose case. For a description of the procedure.3 Analysis of Skin Penetration 1. and finite and Ross 1974): dose case (Fig. related to the permeability coefficient is obvi.5b). 2009). 1. 1999. For the finite dose scenario.4  First five roots of the transcendal Eq.5 shows theoretical skin-concentration so-called transfer coefficient kt with units of per. As for permeation experiments. The used input parameters correspond to the simulation param- He used the early flux of the linear part of the eters used to produce the permeation curves in curve to estimate the fraction of absorbed dose Sect. depth profiles for the infinite dose case (Fig. 2013b) yielding a high variability in fitted parameters. the kinetics ously not possible. Selzer et al. reaches a fulness of this parameter. Values fitted by logistic regression to tableted values from Crank (1975) about the distribution of solute inside the barrier over time. 1. the amount will be heavily overestimated transition into a straight line when steady state is when the flux drops and leans toward zero. the donor.1 Skin-Concentration Depth Profiles Besides permeation profiles (accumulated mass over time) which provide valuable information about the absorption process and the transport to the blood circulation and deeper tissue. Obviously. skin-­ Fig. Scheuplein defined the Figure  1. as shown by Hahn et al.5b). comparable kinetics at short times. The semi-infinite case shows two major drawbacks tremendously limit the use. 1. we representation can overcome ambiguities and kindly refer to Chapter 10. 1.5a).5a).3. Sect. Melero et al.25 for concentration depth profiles supply precious data continuous values of β from 0 to 5. Wagner 2004. ences (Escobar-Chavez et al. 1. However. finding a parameter et al.31) (Scheuplein semi-infinite dose case (Fig.31) show the change of concentration over time and specific dose space in the stratum corneum. These curves were pro- duced with the help of the DSkin software and flux ´100 kt = (1. 1. Root 1. time.

Hansen et al. one can calculate the solutions for the diffusion equation can be concentration inside the barrier at point x and obtained to describe the change of concentration time t (Crank 1975. 1. Eq. Hence. Obviously.5  Theoretical change of concentration in the barrier over time for the infinite dose (a). ÷ . For the infinite dose case. Simulations were performed using the DSkin software line in the concentration over space kinetics is over time and space by using the appropriate ini- typically not reached. 2008). permeation .t ) = K ´ C0 ´ çç ç1 . C0 the tion).å sin ç ÷ exp ç . analytical homogeneous membrane. assuming a As for permeation experiments. ÷÷ (1. retical zero concentration (perfect sink ent diffusivity. and l the macroscopic thickness of conditions). and finite dose (c) cases. D the appar. with ææ xö 2 ¥ 1 æ np x ö æ Dn 2p 2 t ö ö c ( x. and no solute is present inside the the barrier membrane with 0 £ x £ l . 1. barrier at t = 0 . the receptor compartment is kept at a theo- initial concentration in the vehicle. The equa.5c). only the passive dif- tion can only be applied if the donor does not fusion process is modeled. semi-infinite dose (b). 1.1  Basic Mathematics in Skin Absorption 13 a b Infinite dose Semi-infinite dose c Finite dose Fig.11).32) èè l ø p n =1 n è l ø è l 2 ø ÷ø As for the permeation case (Eq. and a significant drop can tial and boundary conditions together with be recognized over time (Fig. K denotes deplete over time (typical infinite dose assump- the barrier/vehicle partition coefficient.  1.3.

profile at a certain time cannot be obtained narios (Selzer et al.. For late times.34) n=0 ( 2 n + 1) p è 2 l ø ç è 24 t ÷ lag ø membrane thickness provides the area under skin- concentration depth profile curve (AUC.35 can be useful to predict the AUC the diffusion equation can be obtained with α and as a function of time after obtaining K and 2 D β (as defined earlier in Eqs. The AUC equals the total amount of the period of delay before the stratum 6 Dcorneum is drug present in the membrane at time t divided by stripped with td = t . 2008) or even other exposure sce. Selzer et al. immediately at the end of an experiment (e.2 ç2 p å n = 0 ( 2n + 1) 2 exp ç - ç l 2 ÷÷ ÷÷ (1.35). simplified formulation of Eq. 2011).32 across the 2006): æ1 4 ¥ 1 æ ( 2n + 1)2 p 2 Dt ö ö AUC = K ´ C0 ç .g. fitting Eq. and the skin-­concentration depth profile can be described æ xö css ( x ) = K ´ C0 ç1 .25 and 1. 1.Obviously. only be obtained before the steady state is Eq.37) n =1 b + b 2 + a n2 è l ø . an analytical solution for Hence. the membrane can be easily calculated from a netics for the evaluation of topically applied com.2 ÷ (1.35) è è øø In 1998. enhancement.t ) = 2 ´ K ´ C0 ´ å exp ç. 2007).32 to experimental data should D coefficient ( 2 ) and extrapolate the skin-­ focus on short incubation times. 1. l concentration depth profile for later time points If experimental data that account for the depth (Naegel et al.÷ (1. Naegel et al. For the finite dose case.a n sin (a n x / h ) æ a n2 Dt ö c ( x. binding phenomena. the Food and Drug Administration For the steady state. the profile will reach steady state. the amount of solute in released a draft guidance on dermatopharmacoki. 2013a. and Eq.24) from parameter fits (Herkenne et al.( 2n + 1)2 p 2 t ö c ( x. Hence.33) with è l ø ¥ 8 æ ( 2n + 1) p x ö æ . and convec. 1. M ss = A ´ l ´ (1. Integration of Eq. l2 Here. the skin-concentration depth due to a long tape-stripping procedure). 1. in order to obtain kinetic param- values for partition coefficient (K) and diffusion eters. b. of exposure. Besides the maximum K ´ C0 amount in the outermost skin layer (stratum cor.35: pounds (Shah et al. texp denotes the duration the volume of this compartment (Herkenne et al.obtained values have to be adjusted according to plifies to the time delay (Reddy et al.36) neum) and the time to reach the maximum amount. 1998). 1. 1.texp .32 sim. information about diffusivity can tion effects are not part of the model. tlag is the lag time with tlag = .14 D.  1.32 can be of great benefit to estimate the reached. and td is Eq. l (Kasting 2001): ¥ b cos (a n x / h ) . Eq. 1. 2000b).t ) = K ´ C0 å cos ç ÷ exp ç d ÷ (1. 2 the AUC was defined as a parameter of interest.

1. dt (1. Systemic circulation b e.38: A. and V is the the number of compartments used to describe the skin bar.38) often be derived with little hassle in dt ­comparison to complex multicompartmental a e. two-compartment (b) skin model.k-1C2 + k-2 C3 . rather than the number of overall compartments. the solution of a set of ODEs can dC2 V2 = k1C1 .39 and hold (e.6  Exemplary sketch of a one-compartment (a) and C denotes the average concentration in the compartment.k2 C2 . Eq.g.g. volume of the compartment that is accessible for the solute rier.k-1C2 + k-2 C3 .1  Basic Mathematics in Skin Absorption 15 As for the infinite dose case. For stiff equations. Eqs.k-2 C3 + k-3C4 .40) Compartmental or pharmacokinetic models (PK) are used since the early beginnings of mathemati. respectively: in the barrier at t = 0 ). Vehicle e.39) 1.3.k2 C2 (1. Vehicle e. 1. heterogeneity of the skin (McCarley and Bunge mal drug absorption. for obtain a series of ordinary differential equations example.2 Skin Compartmental Approaches dC3 V3 = k2 C2 . 1992). a set of first-­ tiation formula (BDF) is a well-known algo- order rate constant can be assigned to denote the rithm.. perfect sink conditions and no solute 1.g.2) to Numerical solvers for nonstiff equations are. Obviously. Here. Stratum corneum e. only for easy equations) or numerically. distribution . The separation of the skin in a lipophilic part cal description to study the fate of a substance (the stratum corneum) and hydrophilic part (via- that was applied in the systemic circulation and ble epidermis) is common practice to mimic the are also part of the history of describing transder. the same limitations For the two-compartment model. using compartmental models can have one-compartment model can be expressed by some advantages. The underlying ODEs series of well-stirred compartments. model and a two-comparment skin model. Systemic circulation Fig. dC2 V2 = k1C1 . Euler’s method or the famous fourth-­ (ODEs) that describe the change of amount of order Runge–Kutta method (widely known as solute in different compartments over time. RK4). They treat the body as a 2001. Viable epidermis e. the number denotes assuming simplified well-stirred conditions. partial differential diffusion equation (Eq. Seta et al.g. Commercial and free mathematical transfer of a compound from one compartment to software package usually provide various possi- another.k3C3 dt (1. Skin e. Figure 1.g.g.g.6 shows two examples of skin bilities for the convenient solving of these kinds compartment models: a one-comparment skin of equations. The basic can be solved either analytically (for the most idea is the elimination of space dependence of the part.40 hold. 1. the backward differen- For the family of PK models. 1. In comparison to solutions of the diffusion The corresponding first-order ODE of the equation.g.

2000a. (Hadgraft 1979. in 1992. 2001. Despite a growing trend toward numerical dif. Reddy et al.4. approaches (Neumann et al. Guy et al. we presented basic mathematical can be implemented very easily in compari. Skin Absorption B. Complicated exposure scenarios with. 2013). The interested stants to physicochemical parameters of the dif.1 Laplace Domain Solutions diffusion models (Hansen et al. in the time and/or space domain (see Sects. fusant and fit experimental plasma curves or Fitzpatrick et al. to the time domain. more complex exposure scenarios on a macro- fication of having well-stirred compartments that scopic and microscopic scale. Another popular approach to solve the diffusion fusion models. Laplace solutions and calculated the flux between each element. concepts to analyze skin permeation and penetra- son to complex diffusion models. diffusion models which are more ­cumbersome 1. o ( 1. (such as QSAR models (Potts and Guy 1992. sums and can be expressed for various exposure mental models. but most mathematical or . Even numerical integration of the ODE is and 1. conditions with relatively simple adaptions.t ) exp ( − st ) dt ^ ences (McCarley and Bunge 2001. In comparison to analytical partment into several hypothetical thin elements solutions in the time domain. this section. Statistical models obviously does not reflect reality. 1. In sion equation numerically.4 Advanced Mathematical ­mathematically. Approaches for Studying lytical solutions can be derived with ease.2). They showed that an efficient skin com. 1. typically restricted to making predictions are 1982). partmental model can be achieved by a two-layer Generically. Certain kinetics. 2002. 1980. These models typically relied C. 2004. Adding systemic PK models to a skin PK on analytical solutions of the diffusion equation model can be achieved with little overhead. McCarley and Bunge 2000). periodic application or evaporation Previously. the Laplace transform L is defined as the interested reader is kindly referred to refer. but rather trends toward numeri. For excellent overviews on PK models. Guy and Hadgraft 1982). much more easily.s ) = L ( C ( x. even ana. for example.16 D. approach. Selzer et al. of the diffusion equation typically lack infinite This approach can be numbered among compart. viable differential equation (ODE) which can be solved epidermis. Agatonovic-­ rily. For simple models.5. gen- erally with less success than that for the class of 1. beyond the scope of this chapter.4. 1. Seta et al. A lot of effort was spent to relate rate con. cal finite difference approaches which we will A drawback is the need of (numerical) inversion present briefly in Sect. 2007)) that are analyze in vivo data (Kubota 1991. tion experiments. 2005. ∞ C ( x.1) or used simple compartmental simpli- typically much faster than solving the diffu. (1992) presented another compartmental approach and The huge advantage of solving partial differential successfully studied the transport of radiolabeled equations (PDE) with the help of Laplace trans- hydrocortisone through hairless guinea-pig skin forms is the reduction of a PDE to an ordinary using three skin layers (stratum corneum.41) Besides PK models. Guy and Hadgraft 1985). and dermis). fications to represent the skin (see Sect. we will briefly discuss more advanced models that are capable of describing One big drawback comes with the oversimpli. PK models were typically used in the past to Kustrin et al. 1995) or statistical learning characteristics can only be described unsatisfacto. of the Laplace transform to an objective function (2011). for a concentration function C(x.4. such as the transition of first-order to zero-­order Wilschut et al. t). Chen et al. They divided each com.3 D. reader is referred to references (Patel et al. Neumann 2008) in vitro permeation profiles to PK models (Wallace and Barnett 1978.t ) ) = ∫C ( x. the class of PK models recently equation for various scenarios is the application regained attention by the work of Davies et al.

The discretization allows the construction of essary input parameters (for the simple diffusion coarse-grained macroscopic and fine-grained problem. Figures are not drawn to scale (e. typically much more convenient to relate the nec. compiled a large amount of Laplace domain solutions of the diffusion equation for different scenarios in reference (Anissimov et al. The basic (Kubota et al. see Eq. in Fig. Anissimov et al. 1. These Macroscopic models do neglect the heteroge- models can be used for descriptive and predictive neity of different skin layers and use effective tasks as well as for the theoretical investigation of kinetic parameters of a diffusant for each layer to . for the most trivial form.g. Solutions for special cases of variable partition and/or diffusion coefficients inside the stratum corneum are available in the Laplace space (Anissimov and Roberts 2004). 2002). Frasch fruitfully applied a Laplace domain solu- tion from Anissimov and Roberts to simulate the Macroscopic skin model permeation of theophylline through the stratum b corneum incorporating slow binding phenomena (Anissimov and Roberts 2009. DLip KLip Even the effect of desquamation could be investi- DCor KCor gated. Microscopic SC model 1. delivered Laplace domain solutions for different epidermal turnover rates (Simon and Goyal 2009). and a microscopic representation of the brick-and- mortar structure (Elias 1983) of the stratum corneum (SC) tions are important techniques and allow a more with diffusivities and partition parameters for the lipid and flexible construction of advanced skin models the corneocyte phase (b). 2013). 1990). and Simon et al. Frasch et al. controlled release vehicles idea consists of the discretization of time and (Kurnik and Potts (1997).. 2013). a forming this task easily and in a convenient man- ner. Some important parameters. 2001). microscopic models (a basic example is depicted cients are needed) to physicochemical parame.. b)). 2011).2). only diffusivities and partition coeffi. 1. Finite dosing and finite receptor and variable receptor clearance for the infinite dose scenario could be successfully modeled by Anissimov and Roberts (1999. 2013a. iontophoretic transdermal delivery (Wearley et al.g. 1.2 Numerical Diffusion Models Fig. repeated applications the interplay among input parameters.7). such as steady-state flux and lag time can even be extracted from the Laplace solution without inversion (Anissimov and Roberts 1999).7  Simplified depiction of a macroscopic diffusion Numerical methods for solving the diffusion model with effective diffusivities and partition coefficients equation for various initial and boundary condi. ters of the diffusant (Hansen et al. In comparison imation of the partial differential diffusion equa- with classical PK skin compartment models. (a). incorporation of binding effects (George 2005)). and in vitro lateral space domain (gridding) and a numerical approx- transport (Selzer et al. it is tion (e. For further reading.4.1  Basic Mathematics in Skin Absorption 17 s­ cientific software packages are capable of per.

18 D. which decrease the (an optimized form of Gaussian elimination that number of computational steps in spite of the . shown that this explicit method is only stable for niques exist to approximate the solution of the values of M £ 0. these models typically require Because of the fact that every point of a certain much higher computational costs in comparison layer only depends on points from the past. always does for the diffusion equation) in nons- fusivity.43) representation of the stratum corneum (Hansen et al. Assuming a constant dif. A violation of this recommendation can probably the most prominent method is the finite produce noise which may result (empirically. It can be From a numerical perspective. 1. it is possible to solved by.7b) and hence yield a more in-depth analy. to coarse-grained macroscopic models. with sis of the underlying transport processes.44) (∆ x) 2 the definition of diffusion parameters but also on the definition of spatial geometry. Selzer et al. The left-hand side of equation is approximated An elegant way to overcome this problem is by first-order forward differences and the right-­ by the use of implicit methods that rely not only hand side by second-order central differences on data from the last time point. of numerical instabilities that depend on the pic diffusivity in the stratum corneum) (Muha value of M (Eq. can solve tridiagonal systems of equations).45) ∆t 2  ( ∆x ) 2 ( ∆x ) 2   This equation yields a system of linear equa. For the one-dimensional case. see Fig. the time and space Crank-Nicolson method (Crank 1975) (Eq. 1. the above-­mentioned underlying diffusion equation for the one-­ equation can be easily generalized to problems in dimensional case and hence treat the skin as a 2D and 3D space. in the stratum corneum). it differences approach. In this ∆t case. case). there is no functional relationship between dif- ematical homogenization. for example. Cit denotes the that is unconditionally stable and provides a bet- concentration at point i × ∆ x for time t × ∆ t .5 (for the one-dimensional diffusion equation.42) restriction can lead to extremely small time ∆t ( ∆x ) 2 steps and therefore to high computational runtimes. yields Microscopic models allow a heterogeneous Cit +1 = MCit-1 + (1 . Undoubtful. the absorption kinetics do not only rely on M =D (1.44) and hence on the choice of et al..2 can be approximated by moothable rounding errors. Eq.. anisotro. This leads to the problem preservation of model properties (e. In this case. describe the transport process. tions (tridiagonal matrix) that can be efficiently Using this implicit method. Usually.g. these with a system of constant space step size Δx and kinds of models are only needed for solving the time step size Δt. 2009) (lipid phase and protein phase. the Thomas algorithm choose larger time steps.42 series of multilayered slabs (Fig. Obviously.g. For small diffusion coefficients (e. it is possible to reduce ferent points of one time layer (it is a so-called a microscopic model to a macroscopic one with uncoupled problem). 1. Δt for a given spatial resolution Δx.7a). Reformulation of Eq. As most widely used approach is the so-called for every numerical method. several tech.2 M ) Cit + MCit+1 (1. ter overall error than the explicit approach: Cit +1 − Cit D  Cit−+11 − 2Cit +1 + Cit++11 Cit−1 − 2Cit + Cit+1  =  +  (1. 1. this Cit +1 − Cit Cit−1 − 2Cit + Cit+1 =D (1. By math. Due to the high spatial resolution.45) domain is discretized. 2013b). the that both arise from Taylor series expansions. 1. Selzer et al. 2010. 1.

the skin type. and anatomical site of the urine will represent the rate at which it pene- application (for more information.1  Basic Mathematics in Skin Absorption 19 fact that the solving of the system of linear data can be used for the purpose of establishing equations is computationally more complex ­ IVIVC. others the exposure conditions (such as dose. humidity. sion by time gives the absorption rate (Feldmann Several different endpoints can be used to relate and Maibach 1969. and ethical constraints. George 2005. it is possible for in the field of mechanical engineering and finite ­transdermal patches which act systemically to volume approaches that can efficiently handle demonstrate BE based on plasma levels or unstructured girds and is often applied in the field ­excretion data. if one assumes that the steady-state graph should give an overview on which kind of flux through skin in vitro (Jss [μ g/cm2/h]) is .v. 2013b). Results are harmonized protocols should be compared. physiological. urine). the transdermal Naegel et al. 1989. (Barbero and Frasch 2005.5 I n Vitro–In Vivo Correlation and Maibach 1985). and exposure step. At the same time. Consequently. the rate at which the compound appears in duration). temperature. 2002. Heisig et al. infusion. a transdermal patch of computational fluid dynamics are known to provides an infinite source and controls the inva- successfully model transdermal transport sion rate into the blood similar to an i. In particular. erally demonstrated based on a pharmacokinetic Finite differences are not the only mesh-based study comparing the plasma levels of test and ref- technique to find approximate solutions to partial erence product. 1972. Wester and Maibach 1976). Factors which are known to Maibach 1969. Franz et al. ­pharmacokinetics are correlated to the pharmaco- The method of finite elements that is widely used logical effect. and exact information on relevant kinetic data. for example. 2013). 1974). if skin penetration is the rate-limiting vehicle. Kubota et al. This para. divi- mental. tivity labeling as a technique to obtain pharmaco- cumstances. Finite dosing is of course more relevant to the real exposure to chemicals or actives. Naegel et al. form excretion to the experimental conditions is frequently lacking urine in human volunteers (Feldmann and (ECETOC 1993). Similarly. excreted. a correction for the proportion of uri- Therefore. and conse- quently also for demonstrating IVIVC (Bronaugh 1. 2008. Nitsche forms. following the idea that the differential equations. in vitro to in vivo skin absorption data. For systemically acting drugs. 1996. in this case. Likewise. the be correlated with the plasma concentrations interested reader is kindly referred to references (Chien et al. only data which were obtained using nary to fecal excretion is required. Various works The topic of IVIVC is especially relevant for on modeling transdermal transport that are based estimating bioavailability (BA) and for bioequiv- on finite differences exist (Kurnik and Potts alence (BE) testing of topically applied dosage 1997. Indeed. Feldmann and influence transdermal absorption include among Maibach 1974. tion area and incomplete urinary excretion. as the diffusion equations. For solutes which are also fecally reader is referred to Chapter 10 in this book). compared to the explicit method. are excreted over time and corrected for applica- ies is usually quite different mostly due to experi. 2011. For absorption rates which are measured in vitro can excellent overviews about numerical models. Selzer et al. Bartek et al. 2009). the interested trates the skin. The calculated as percent of the applied doses which type of data obtained in in vitro and in vivo stud. (Frasch and Barbero 2013. BE is gen- and Frasch 2011). In (blood. accurately (IVIVC) detecting the low systemically absorbed and excreted amounts of solutes which are applied to In vitro–in vivo correlations (IVIVC) address the the skin as a finite dose requires sensitive and question of the significance of in vitro data for selective analysis in complex biological media predicting the situation in the living being. Data which were obtained during vitro data are often compared to in vivo data from the 1960s–1980s therefore often report radioac- studies obtained under different experimental cir.

noticed that dif- the joints. reported that the in vitro plasma levels accordingly (Franz et al. 2000). requirement for high number of test persons.v.. achieved with a developed dosage form to obtain In fact. Franz et al. ensure comparability between in vitro and in vivo Difficulties arise for topically (i.nized (Franz et al. a finding which was also supported dynamics (Opdam 1991. later in a follow-up study. 6 h (Treffel and draft guidance document recommending tape-­ Gabard 1996. Fisher et al. during method for in vivo BA or BE testing (Franz et al. regional variances. The rea- ies (with known problems such as poor sensitiv. such ity. method was able to discriminate between differ- Cnubben et al.e. and Cl reader is referred to references (Herkenne et al. for example.sons for these differences can be manifold. 2003). due to issues dose (2 mg/cm2 vs. 2011). 1985) to calculate an in vivo ference was not picked up by the vasoconstric- percutaneous absorption rate (μ g cm−2h−1) based tion assay (Franz et al. From between in vitro and in vivo studies were harmo- this. Russell and Guy 2009)). they derived in vivo permeability coeffi.incubation times (Wagner et al. that is. In particular. such as et al. 2009). In vivo and stripping for demonstrating BE of topically act. Css [μ well as in vitro (for more details. roidal anti-inflammatory drug flufenamic acid This results in the unfortunate situation that the in vitro and in vivo by tape-stripping. Comparing the skin penetration of the nonste- get the stratum corneum itself (Narkar 2010). while this dif- 1978. 2002). 2009). provided that the study protocols i.which were obvious in tape-stripping experi- fiable nor relevant for their local activity.e.v. Wagner in vivo skin blanching or vasoconstriction assay et al. and consequently high costs). Lehman et al. 3 mg/cm2). application. or dermatopharmacokinetics after application could in vitro not be detected (DPK). equivalent to the rate of input to the systemic cir. on or in the results. or cartilage) acting drugs for ferences between several sun protection products which the plasma concentration is neither quanti.in vitro protocols differed in terms of the applied ing drugs was withdrawn in 2002.46) measurements which can be performed in vivo as Here. Unfortunately.. With Css and Cl being 2008. Second. Treffel and Gabard as well as Chatelain skin) or regionally (i. observed that the in vivo levels were consis- (only for corticosteroids) is currently the only tently lower than the in vitro levels which they FDA-recommended assay for demonstrating BE found inside the stratum corneum after different of generics which does not rely on clinical stud. the interested g/l] = steady-state blood concentration. for example. Selzer et al. as interindividual differences. will be more relevant. tape-stripping is only rec. have different pressures applied during tape-stripping . measuring the drug concentration at surements in human volunteers as early as 30 min the site of action. muscle. Currently. these are known. collected substantial evidence that in vitro skin culation in vivo. these are the already mentioned tape-stripping DPK J ss ´ A = Css ´ Cl (1. then in vitro percutaneous absorption testing may be used as a surrogate absorption data can be used. such as for antifungals that tar. two types of measurements zation based on known clinical data: are being discussed for this purpose. First. 2009. from i. Chatelain et al. a until later time points. The differences arising from poorly defined experimental stan. Vaughan and Dennis by differences in clinical efficacy.may be also explained by artifacts from nonphys- dardization. as well as others demonstrated excellent utes after dermal application and after a reference IVIVIC.also permeability measurements which are per- get flux can be calculated which needs to be formed in vitro. further used linear systems ent vehicles. in nearby tissues. Importantly. a tar.et al. [l/h] = systemic clearance. 2009). cients and lag times and compared these to values The latter aspect is certainly very important to obtained in vitro.20 D. administration (Cnubben et al. Franz on the plasma concentration–time profile of sol. A [cm2] = area of skin.iological swelling of the skin inside the Franz dif- ommended by the FDA for BE testing for certain fusion cell obscuring early effects. formulation development for formulation optimi. For ments as well as sun protection factor (SPF) mea- those drugs. Franz et al.. classes of drugs.

implementing more complex 2002).. 1. This collection of reports shows that functions and fitting to experimental data can be obtaining a good IVIVC is possible. Some they demonstrated an excellent IVIVC (Wagner researchers only use <10 summations.2 Fitting of Experimental Data guide when conducting mathematical analysis of experimental data.6.g. To achieve maximal accu- after undergoing abdominal reduction surgery racy. fitting of experimental data to analytical solutions of the diffusion equation is an excellent way of analysis. Despite this fact. and with Hi(x) going to zero for i going to infinity. ple. Hence. the authors which these parameters are obtained are most used the popular scripting language Python2 in critical. difficulties when computing analytical solutions of the diffusion equation and fitting these solu- tions to experimental data. 2008). 1. the authors support the approach of individual These are typical examples of solutions of the evaluation of experiments and averaging of indi- diffusion equation for various scenarios.6 Tips and  Tricks These kinds of packages provide predefined functionalities for root-finding. sum has to be truncated at some point. F ( x ) = a åH i ( x ) + b. For data evaluation.org 2  . To exclude interindividual differ. To gain confidence. correlating the From a practical point of view. where they used abdominal nificantly contribute to the overall result (Frasch skin from the same exact individual before and and Barbero 2008). However. such that they could show excellent agreement of the drug 1 + e 0 > 1 holds for a certain machine. However. (1. mean accumu- ¥ lated mass over time). 2001) to compute various equations presented for the finite dose case. As reported here. For example. and This section supplies information about common plotting. combination with the SciPy package (scientific python) (Jones et al. Being vidual determined parameters. like the so-called obtained from tape-stripping were not usable due machine epsilon of the computer – a relative error to the disinfectant wipe which was necessary for due to rounding in floating point arithmetic the surgery and which significantly changed the (Hansen et al. since the subsequent terms do not sig- a special workflow.1 I nfinite Sums in Analytical experiments are typically conducted in a recur- Solutions rent manner.47) It is strongly recommended to report the eval- i uation of single experiments (OECD 2004b). Wagner et al. 1. dataset (of every single experiment) or a combi- ter include an infinite sum of form nation of experimental data (e. Nonetheless. Python: http://www. Unfortunately. The machine epsilon ε0 is drug levels inside the upper skin layers. Nowadays. defined as the smallest number.6. It should work as a 1. the infinite amounts found after different incubation times. able to deal with this kind of equations is a key many researchers prefer to handle arithmetic requirement for a sound evaluation of skin trans.python. researchers can fit the objective function to either each individual Many analytical solutions presented in this chap. designed Sect. for the infinite dose solution presented in ences in a follow-up study. we will give a few hints for the solid handling.1  Basic Mathematics in Skin Absorption 21 to name but a few.1. 2000). a more precise way to truncate the sum is to (Wagner et al.3. the results incorporate a relative threshold. fitting. levels inside the deeper skin layers (Wagner et al. achieved easily using commercial or free data the parameters which should be correlated need analysis and statistical software packages or free to be carefully selected. 2002). for exam- et al. and the conditions under scripting languages. means of combined experiments (Schäfer-­ port experiments.

every i. Finite vehicle volume and solvent deposited solids. improve our understanding of skin absorption.. Effects of a additive in nature (Selzer et al. Frasch F (2009) Pig and guinea pig skin as once (Kasting 2001). New York. Maibach HI (1972) Skin perme- tal data. These were the so-called weighted root mean square function highlighted in this chapter. Raykar PV (1988) ble of emphasizing or de-emphasizing certain Heterogeneity effects on permeability–partition coef- data points. a of percutaneous absorption kinetics. surrogates for human in vitro penetration studies: a quantitative review. for fitting accumulated mass versus time data tum corneum depth profiles and desorption kinetics. 2009). molecular descriptors. In: Walters KA including toxicology. and viable epidermal s resistance for a constant donor concentration. pp 197–270 . Key to the useful application of Marcel Dekker AG. This requires knowledge about the Sci 88(11):1201–1209 kind of underlying error or variation.g.22 D. Roberts MS (2004) Diffusion modeling ti of percutaneous absorption kinetics: 3. predict relevant endpoints. Jepps OG. J Pharm Sci 93(2):470–487 for the infinite dose case. 2007). minimization of mental and model constraints.48) References Agatonovic-Kustrin S. The basic idea is to minimize the sum biological mechanistics but also of experi- of squared residual errors (e. LaBudde JA.dicalc ) n i =1 (1. This can be helpful for exam. Roberts MS (1999) Diffusion modeling To emphasize experimental uncertainties. (ed) Dermatological and transdermal formulations.48): 1 n RMS = åwi ( diexp . J Invest Dermatol 93(2):280–286 weights for every di pair. Raykar PV (1989) Solute structure–perme- in the acceptor compartment). 2013a). Selzer et al. Roberts MS (2013) the partition coefficient). diexp is the ith ANN modeling of the penetration across a polydimeth- ylsiloxane membrane from theoretically derived data point. drug delivery. 2008). 1. Bartek MJ. Toxicol In Vitro 23(1):1–13 Conclusion Barbero AM. Ann Biomed Eng 33(9):1281–1292 hands which allow us to interpret experimen. Effects of flow 1 typical choice of weights is wi = 2 (Henning rate. tory affairs. consequences for stra- ple. suggested a weighting scheme 1 90(4):504–520 with wi = to de-emphasize later time points Anissimov YG. J Pharm et al. These weights are capa. and ability in vivo: comparison in rat. Pharm Res 5(9):566–573 Anissimov YG. If no weighting is applied. Yusof APM (2001) Here.g. dermal and dermal transport processes. Higuchi WI. Roberts MS (2009) Diffusion modelling that the standard errors of later time points are of percutaneous absorption kinetics: 4. receptor sampling rate. displayed in Eq. Walters KA. dicalc is the quantity that is being mod. Watkinson AC (2002) Methods They find their applications in various fields. wi is 1 for ficient relationships in human stratum corneum. slow equilibration process within stratum corneum on Fitting finite dose permeation data can be absorption and desorption kinetics. Variable diffu- (Krüse et al. or regula. and wi are optional ability relationships in human stratum corneum. J Invest Dermatol 58(3):114–123 Brain KR. Frasch HF (2005) Modeling of diffusion with partitioning in stratum corneum using a finite ele- We hold powerful mathematical tools in our ment model. Adv Drug stant value and/or fit data from several doses at Deliv Rev 65(2):169–190 Barbero AM. Krüse et al. sion and partition coefficients. J Pharm Sci data. for studying percutaneous absorption. Anderson BD. A stabilization can be Mathematical and pharmacokinetic modelling of epi- achieved by either fixing an unknown to a con. Beresford R. pig and man. since it is well known Anissimov YG.. concentration in the membrane at time 241–254 t and point x or cumulative amount of substance Anderson BD. Korting et al. n is the number of data points. Roberts MS (2001) Diffusion modeling standard deviation is not the only option to weight of percutaneous absorption kinetics: 2. Fitting is typically done by such mathematical tools is a fundamental applying a linear or nonlinear least-squares understanding of the underlying physical and approach. J Pharm Sci 98(2):772–781 prone to variabilities in the unknowns (especially Anissimov YG. rabbit. J Pharm Biomed Anal 26(2): eled (e. Using the Anissimov YG. 1. Dancik Y.

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1    Introduction  27 2. University of Tanta.4. Occlusion can increase the water content of the skin. 2. H. College some systems such as Transfersomes was linked of Pharmacy. Taking into consideration the fact that water acts as skin penetration enhancer. occlusion cannot be taken as the only tool to enhance transdermal drug delivery. Occlusive Versus Nonocclusive Application in Transdermal Drug 2 Delivery Gamal M. the efficacy of Department of Pharmaceutical Technology. or Delivery Systems  30 transdermal therapeutic systems (Zhai and Conclusion  32 Maibach 2002). Occlusion can be obtained from References  32 the occlusive nature of the formulation itself.4. with lower efficacy on polar drugs (Cross and Roberts 2000.I. Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin. Maibach (eds. El-Geish Street. wound dressings. Treffel et al. occlusion can enhance percutaneous absorption of drugs (Barry 1987). gloves.2 Supersaturable Transdermal diapers.1 TransfersomesTM  29 impermeable films or others. El Maghraby to the nature of the delivery system. 31111 Tanta.M. textiles garments.2    Permeability Barrier of Human Skin  28 Dermal and transdermal drug delivery can be achieved after occlusive or nonocclusive (open) 2.1 Introduction 2.4    D  elivery Systems Utilizing the Method of Application  29 Occlusion can be achieved by covering the skin by 2. occlusion was shown to enhance the permeation of lipophilic drugs. such as tape.com The following sections will provide a brief over- © Springer-Verlag Berlin Heidelberg 2017 27 N. For example. 2. oleaginous formulation will have strong occlusive nature (Berardesca and Maibach 1988). El Maghraby Contents 2. 1992). However. Dragicevic. With regard G. With respect to the nature of the drug. This is due to the complex nature of percutaneous drug absorption and its enhancement which depends on many factors including the nature of the drug and the transdermal drug delivery system. DOI 10. e-mail: gmmelmag@yahoo.1007/978-3-662-53270-6_2 .). Egypt with open application (Cevc and Blume 1992).3    E  nhancing Effect of Occlusion on Transdermal Delivery  28 application with the former being traditionally considered as the standard application technique.

1.28 G. These lines of defense 1. drug permeation have been identified as the tran. occlusion does not the keratinocytes and across the intercellular lip. The advancement in research methods resulted in clear and detailed identification of skin structure and its barrier function (Scheuplein and Blank 2. sites that can sequester drugs providing the first In this study. El Maghraby view on the occlusive versus nonocclusive appli. The second defense line is the from a commercially available test ointment closely packed hydrocarbon chains of the inter. The of Occlusion on Transdermal studies were extended to cover the barrier func. drugs pass ing in a loose organization of SC lipids. The former is considered as forms hydration shells around the head groups the main route with the latter playing only minor of the SC lipids. 1988). This can result in over- that the stratum corneum (SC) is the main barrier hydration of the SC with subsequent reduction for transdermal drug delivery. SC lipids providing room for drug permeation cellular lipids. Occlusion was tains minimum amount of water which is achieved by the placement of a piece of high- provided by the hydrophilic domains in the dia. Elias 1981. Two pathways of in the barrier nature of the SC (Bucks et al. the authors evaluated the penetra- difficulty in their penetration through the trans. together with the effect main barrier in both the intercellular and trans.2 Permeability Barrier (Warner et al. Barry 1987). There was a signifi- The integrity of human skin is also linked to cant difference in the epidermal flux of para- its water content which varies across skin strata bens provided by the different vehicles applied . and two commonly employed solvent vehicles cellular lipid lamellae. which considered both the (amphiphilic compound) was the same under intercellular and the transcellular pathways occlusive or open application (Treffel et al. In a more recent study. This leads to the disruption of role in transdermal drug delivery (Scheuplein the closely packed hydrocarbon chains result- 1967).3 Enhancing Effect 1971. SC. 2. addition. but the transdermal flux of caffeine are shown in Fig. tion of a mixture of paraben ester preservatives cellular pathway. 2. which is philic domains of the lamellar structure of the a continuous but tortuous way through the inter. 1991). but increases deeper into the skin where it reaches a five times higher value at the basal skin layers 2.6 times. increase percutaneous absorption of all types ids. the water concentration is less than 15 %. This line provides the (acetone and ethanol). This major pathway con. the effect of in the cells which contains hydrogen-bonding occlusion was shown to depend on the vehicle. It is now accepted loss from skin surface. However. immediately after dosing. and the transcellular route through (Barry 1987).1. In addition to this.M. This has been reviewed and documented Skin occlusion can inhibit the continuous water (Bouwstra and Ponec 2006). For example. Orland 1983. In through the intact SC. occlusion increased the many defense lines which hinder their progress permeation of citropten (lipophilic compound) into and through the skin. the SC con. Delivery tion of the skin in the healthy and diseased state. The diagram represents the keratin 1992). In the transepidermal pathway. servatives from these systems. The penetration-enhancing effect of sepidermal and the transappendageal pathways water can be explained on the basis that water (Scheuplein 1965). Passing through these routes. Barry 1983). excess water can extend the hydro- tains two routes: the intercellular route. This creates a transdermal of Human Skin hydration gradient which is characteristic for normal skin (Cevc and Blume 1992). drugs will face of drugs. with the concentration of water increasing as we cation and how it can alter the transdermal drug move from the skin surface to deep strata. of occlusion on the rate of delivery of the pre- cellular routes. At the delivery. density polyethylene on the application site gram (Fig. (Barry 1987).

carrying therapeutic a less damaging effect on the skin compared to amounts of drugs if applied under nonocclusive the occlusive application of the same solvents.1 TransfersomesTM tion (Cross and Roberts 2000).2  Occlusive Versus Nonocclusive Application in Transdermal Drug Delivery 29 Fig.4. They lower penetration-enhancing effect. open application of these sol. a decreased flux was seen upon the occlusive application of the ointment formula. Open indicated that these vesicles can penetrate into application of these solvents will thus provide and through intact skin. importance of the application method on the vents will lead to their rapid evaporation and transdermal delivery of drugs from the highly hence shorter contact time with the skin and deformable lipid vesicles.1 Simplified diagram showing a model Keratin in cells with for the stratum corneum hydrogen bonding sites with various defense lines against penetration of xenobiotics (Modified from Barry 1987) Closely packed hydrocarbon chains hydrophilic domain under occlusion. TransfersomesTM.4  elivery Systems Utilizing D observed from acetone and ethanol used as the Method of Application vehicles. The ability of such carriers to pene- . method. 2. In contrast. Whereas increased flux was 2. conditions. Cevc and Blume (1992) highlighted the ity. These results can be explained on the basis that occlusion While most of the application studies employ increased the contact time between the skin mainly occlusive application with no attention and the volatile solvents (ethanol and acetone) paid to the possible effect of the application which possess the penetration-enhancing abil. 2.

1992). 1995). Based on this hypoth. The results indi- tum corneal surface being left open to the atmo. sively out of the skin into the formulation. This enhance the transdermal delivery of indometha- was achieved by the developed open hydration cin compared to the corresponding ME formula- protocol in which the epidermal membrane was tion containing increasing concentration of hydrated from viable epidermal side with the stra. with the esis. achieved by the uptake of water diffusing pas- caine entrapped in TransfersomesTM under occlu. Cevc et al. ultradeformable vesicles need to be applied rapid addition of a nonsolvent thereby creating a under nonocclusive conditions. as occlusion is solution in which the drug concentration exceeds believed to abolish the driving force for the skin its equilibrium solubility (Megrab et al. On doing so. 2001b). This the tested ME as indicated from the recorded protocol was employed to study the transdermal transdermal drug flux values which revealed delivery of estradiol from ultradeformable lipo. To test this hypothesis. metastable. acting as nonsolvent. 2000. Taking application of liposomes. through the skin. polyoxyethylene 20 sorbitan Maghraby and coworkers developed an in vitro monooleate (Tween80) and sorbitan monolau- experiment which preserves the transepidermal rate (Span20).2 Supersaturable Transdermal (tendency to avoid dry surrounding). supersaturation can be The same group applied the local anesthetic lido. This can increase the essential property which made the transdermal driving force for transdermal drug delivery. was developed and used to hydration gradient (El Maghraby et al. Vesicles’ which the thermodynamic activity of the drug is deformation ability was considered as another increased above unity. hydration gradient to provide sufficient driving Alternative methods have been adopted to force for vesicle penetration through the skin. The can represent a special case of the first method results showed the superiority of TransfersomesTM and can gain the benefit of occlusive application compared to traditional formulations. supersaturated state in and Blume 1992. This process was used to explain the by researchers in the field of dermal and transder. penetration of vesicles (Cevc and Blume 1992). it was this process into consideration.M. obtain drug supersaturation leading to increased The vesicles were thus given the name “ultrade. El Maghraby trate intact skin was attributed to their xerophobia 2. drug delivery. The . 2002). mixing of cosolvents for a certain drug. Occlusion ME. This the applied formulation (Planas et al. Cevc et al. This superiority was attributed to possible resulted in a significant reduction of the transder.30 G. it was lol after dermal administration using micro- important to test occlusive versus nonocclusive emulsion (ME) (Kemken et al. a self-micro- important to maintain the hydration gradient even emulsifying drug delivery system (SMEDDS) in the in vitro experiments. In the second method. 1999). water (El Maghraby 2010). in which the formulation can mix with skin The above hypothesis was not accepted easily secretion. 1992). in sion using a watertight wrapping for 25 min over which case water can act as the nonsolvent. which Delivery Systems causes the vesicles to resist dehydration at the skin surface by moving into the skin along with Supersaturation is the process of creation of a the local transdermal hydration gradient (Cevc transient. chemical potential and enhanced transdermal formable vesicles” (El Maghraby et al. greater transdermal drug delivery from somes after their occlusive and nonocclusive SMEDDS compared to that obtained from application (El Maghraby et al. b. enhanced pharmacodynamic effect of buprano- mal drug delivery. To verify this. El of ethyl oleate.4. cated the superiority of SMEDDS compared to sphere. to mimic the in vivo conditions. 1995). SMEDDS with the hydroalcoholic receptor tance of the application protocol for the transdermal which will diffuse from the receptor upward drug delivery from ultradeformable vesicles. supersaturation of the drug after dilution of the mal flux of the drug which confirms the impor. The first method was based on 2001a.

associated with linear reduction in the solubility For EME. The ME system was used neat or with lations containing volatile components as candi- ethanol as a volatile cosurfactant. the same method was utilized to for. of the volatile solvent after open application. This method was utilized to significantly higher than that obtained with enhance the transdermal delivery of a lipophilic EFME. increasing the con. It was In a more recent study. the same formulation. The authors introduced ME formu- and water. this cannot be considered as the only taining oleic acid.2  Occlusive Versus Nonocclusive Application in Transdermal Drug Delivery 31 author supported this claim by comparing the transdermal delivery of indomethacin from SMEDDS with that obtained from supersatu- rated microemulsion system which was pre- Solubility (mg/ml) pared by diluting saturated drug solution in s SMEDDS with water which acts as the nonsol- vent. Ethanol-containing centration of the volatile solvent in the formulation ME (EME) was shown to be more effective than usually leads to exponential increase in drug solu. 26–28 % compared to occlusive application of The challenge here is to maintain the supersatura. Open applica- bility in the formulation. 2. This suggests a role for supersaturation model drug (lavendustin). 2. skin penetration enhancement. Later. leaving the drug at a concentration much Fig. This study revealed enhanced transdermal The sum of fluxes obtained from ETW and EFME delivery of testosterone from the spray. propylene glycol. TweenTM 80. Results of this dates for the formulation of sprays for dermal study (Fig. progesterone from 40 % ethanol in water (ETW). 2. tion of EFME produced a minor change in drug tile component after open application will be flux compared to occlusive application (Fig.2). with supersaturation with a contribution of ethanol in the recorded shown to be a promising strategy (Moser et al.2  Exponential solubility profile of a given drug in greater than its equilibrium solubility in the the presence of increasing concentrations of volatile sol- remaining nonvolatile part of the formulation vent. To verify this. mechanism. the 2001). open application reduced the flux by creating transient supersaturation state (Fig. However. thus concluded that the penetration enhancement ery of progesterone from ME was studied under induced by supersaturation played an important occlusive and open application (El Maghraby role in enhanced transdermal drug delivery. authors evaluated the transdermal delivery of mulate a testosterone spray (Leichtnam et al. The linear plot indicates the theoretical solubility of the drug in a supersaturated system created by evaporation (Leichtnam et al. 2. ous propylene glycol (control). 2. 2007). In this case. Evaporation of the vola. The third method for obtaining supersaturated so systems employs rapid evaporation of one or more Concentration of volatile solvent volatile vehicle components upon application to the skin.2 and requires open by So produces the degree of supersaturation application of the formulation to the skin surface. creating supersaturated system before occlusive application to the skin. This study tested a ME formulation con.3) indicated that ME formulations application. Dividing S cally illustrated in Fig. The results revealed similar flux from the SMEDDS and the prepared supersaturated system. 2007). the ethanol-free system (EFME). This process is graphi. Another important requirement of such process is that the drug must have higher solubility in the enhanced progesterone transdermal flux com- volatile solvent than in the nonvolatile components pared to the saturated drug solution in 14 % aque- of the formulation.3). 2012). the transdermal deliv. was lower than that obtained from EME. but the flux remained tion for long time. .

New York/Basel El Maghraby GM (2012) Occlusive and non-occlusive Barry BW (1987) Mode of action of penetration enhanc. El Maghraby GM. ETW is 10 Flux ( µg cm-2h-1) 40 % ethanol in water. J Pharm Pharmacol 1758:2080–2095 51:1123–1134 Bucks D.3 Transdermal 16 delivery of progesterone after occlusive and open Occluisve application application of microemul. Biochim Biophys Acta liposomes: mechanistic studies. tides. J Control Rel 36:3–16 tions in dermal and transdermal delivery of Cevc G. Barry BW (2000) In: Bronaugh RL. EME 12 is ethanol-­containing microemulsion. Guy R. optimization and transfer efficiency in the case of epicutaneously applied pep- the method of application of drug formula. Williams AC. Biochim Biophys Acta standard liposomes in vitro. Richardsen H (2002) Ultradeformable drugs. Schätzlein A. Blume G (1992) Lipid vesicles penetrate into El Maghraby GM. El Maghraby Fig. Schätzlein A. The formulators should consider the lipid vesicles can penetrate skin and other semi-perme- application technique as a factor parallel to the able membrane barriers unfragmented.32 G. 2. Pharmaceutica 80:765–778 ment or drug-like device? Skin Pharmacol 1:207–215 El Maghraby GM. Barry BW (1999) Skin Bouwstra JA. J Invest Dermatol 115:914–918 El Maghraby GM (2010) Self microemulsifying and References microemulsion systems for transdermal delivery of indomethacin: Effect of phase transition. Marcel Dekker. pp 85–114 J Pharm 196:63–74 Cevc G. Barry BW (2001a) Skin intact skin owing to the transdermal osmotic gradients delivery of 5-fluorouracil from ultradeformable and and hydration force. Maibach HI (1988) Skin occlusion: treat. surement. Boca Raton. Biochim Biophys Acta 1564:21–30 ing dermal and transdermal drug delivery Cross SE. and control is 14 % propylene glycol in water (This 8 figure was produced using data published by El Maghraby 2012) 6 4 2 0 EFME EME ETW Control Conclusion Cevc G. somes: refinement of surfactant concentration. Ponec M (2006) The skin barrier in healthy delivery of oestradiol from deformable and traditional and diseased state. J Pharm Pharmacol 1104:226–232 53:1069–1077 . Evidence from double label CLSM experiments and direct size mea- formulation factors (composition) in develop. Blume G (1995) Transdermal drug This overview highlighted the importance of carriers: basic properties. J Control Rel 6:85–97 of progesterone: mechanistic studies. application of microemulsion for transdermal delivery ers in human skin. Int CRC Press. Maibach HI (1991) Effects of occlusion. Williams AC. Maibach HI (eds) Vitro percutaneous Oestradiol skin delivery from ultradeformable lipo- absorption: principles. Scintia Berardesca E. Roberts MS (2000) The effect of occlusion on systems. B Biointerfaces 75:595–600 neous absorption.M. fundamentals and applications. Williams AC. acetone and ethanol vehicles. epidermal penetration of parabens from a commercial allergy test ointment. EFME is ethanol- free microemulsion. Colloids Surf Barry BW (1983) Dermatological formulations: percuta. 14 Open application sion.

Int J Dermatol 20:1–9 absorption. Pharm Res 9:554–558 importance of various routes of skin penetration. Wüthrich P. neous absorption of two compounds with different phys- tion. permeation across human skin and silastic mem. Rolland H. pp 3–63 function. vol 1. Maibach HI (2002) Occlusion vs. Gonzalez P. Blank IH (1971) Permeability of the skin. Transient diffusion and the relative emulsion as vehicle. Muret-D’Aniello P. J Pharm Sci 96:84–92 Physiol Rev 51:702–747 Megrab NA. Myers MC. New York. J Invest Dermatol Orland GF (1983) Structure of the skin. Barry BW (1995) Oestradiol Treffel P. Zhai H. Guy RH (2001) Enhanced Warner RR. tosterone from a spray. Barry BW (2001b) Skin Planas ME. probe analysis of human skin: determination of the rated formulations. Kalia YN. Scheuplein RJ. Williams AC.2  Occlusive Versus Nonocclusive Application in Transdermal Drug Delivery 33 El Maghraby GM. Rodriguez L. Sanchez S. J Pharm Pharmacol tion of local pain insensitivity by anesthetic liposomes. skin barrier Oxford University Press. II. Muret P. Guy RH (2007) J Invest Dermatol 48:79–88 Impact of antinucleants on transdermal delivery of tes. Scheuplein RJ (1965) Mechanisms of percutaneous nization. (1992) Noninvasive percutaneous induction of topical trolled skin delivery of estradiol from ultradeformable analgesia by a new type of drug carrier. Williams AC. Muller BW (1992) Influence of of solubility. Taylor DA (1988) Electron skin permeation of a lipophilic drug using supersatu. J Invest Dermatol 45:334–346 supersaturation on the pharmacodynamic effect of Scheuplein RJ (1967) Mechanisms of percutaneous bupranolol after dermal administration using micro. Kriwet K. J Control Rel 73:245–253 water concentration profile. Skin Res Technol 8:1–6 . and prolonga- and standard liposomes in vitro. Coumes-Marquet S. I. Ziegler A. membranes and kerati. Cevc G hydration and possible shunt route penetration in con. In: Goldsmith LA 90:218–224 (ed) Biochemistry and physiology of the skin. Routes of penetration and the influence Kemken J. absorption. Leichtnam ML. Skin Pharmacol 5:108–113 Moser K. Agache P (1992) Effect of occlusion on in vitro percuta- branes: effects of propylene glycol and supersatura. J Control Rel 36:277–294 icochemical properties. 53:1311–1322 Aneth Analg 75:615–621 Elias PM (1981) Epidermal lipids.

. this can be achieved by regu- larly replenishing the donor solution or.................. Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin. 1995)........ Finite and Infinite Dosing 3 Wing Man Lau and Keng Wooi Ng Contents 3. when examining the fundamental permeation behaviour of a substance or when investigating the effects of penetration enhancers on percuta- neous absorption.. more W...... Brighton BN2 4GJ.... University of Brighton...M....2  Skin Absorption Kinetics........ In practice....... Maibach (eds.. from the solid excess (Megrab et al.... 42 tions are applied to the skin as a finite dose...... DOI 10.....1 Introduction 3...............uk then as the permeant leaves the donor solution.. PO Box 226.. Dragicevic.........lau@reading......... In practice......1 Introduction..................Ng@brighton. permeant to a saturated donor solution... Ng (*) is replaced by molecules entering the solution School of Pharmacy and Biomolecular Sciences..... no change in thermody- namic activity of the permeant) in the donor phase..1007/978-3-662-53270-6_3 . 36 how much of a preparation should be applied to 3.. However........... infinite dosing is usually employed to maintain a constant rate of absorp- tion of the test compound through the skin. often References.......... it is generally assumed that there is no change in permeant concentration within the formulation throughout the experi- ment..2.. With infinite dosing.......... Lau (*) usually...2.......... 36 3.......2 Penetration Kinetics.. more accurately...... 41 the skin surface in order to achieve the desired bioavailability........ assuming Whiteknights.. many topical prepara- Conclusion.... 42 in the form of a cream. gel or ointment. the so-called steady-state flux.....................W...... UK Alternatively... by the addition of a small excess of solid School of Pharmacy..... that is............. University of Reading. infinite dosing is commonly e-mail: K..... When using Franz diffusion cells.. it K..........uk assumed by administering an amount of the © Springer-Verlag Berlin Heidelberg 2017 35 N.. 35 The aim of permeation studies is to determine 3.. this cor- responds to a constant permeant concentration (or.........1 Permeation Kinetics.ac.. H..ac... Huxley Building... Lewes Road.. e-mail: w.).......I. UK that dissolution of the solid is not rate-limiting.. Reading RG6 6AP.

both infinite dose and finite nents. as well as the dura. Under be estimated from the cumulative amount of these conditions.1). 3. a small amount of position or composition of the permeant. 3. the experiment. it may penetrate and diffuse dohomogeneous membrane providing a single rapidly or may bind to stratum corneum compo- transport route. amounts of >100 μl cm−2 or >10 mg cm−2 A typical infinite dosing regimen will produce a are required. A lag phase is seen where the amount of dose experiments can be described by Fick’s laws permeant in the receptor compartment increases of diffusion. Ng f­ormulation large enough to preclude any ‘sig. proportional to the concentration gradient accord. For example. the absorption of a substance into or on the permeants’ physical and chemical proper- through the skin is regarded as a passive diffusion ties (e.W.2) to provide suitable data. as shown in Fig.donor potential). when a permeant is 3. can be barrier. the parameters describing the concentration of permeant in the stratum cor- absorption of a substance through the skin barrier neum. J. According to the Organisation for Economic Co-operation and Development (OECD).2 Skin Absorption Kinetics applied to the skin. p­ ermeant passing through a unit area of mem- nificant’ reduction in the test compound or any brane at time t (Crank 1975): other component in the donor phase during the dc course of the experiment (Franz et al. To determine how dif- Under finite dose conditions. which is fully occupied (or at steady-state equilibrium). 1993).1) these cases. 2006). bind. After a sufficient time (again dependent on (such as the permeability coefficient and lag the nature of the permeant. the employed: experimental conditions should mimic as dc d 2c closely as possible the in vivo situation in order =D 2 (3. J can permeant develops across the membrane. D is the diffusion coefficient and δc/δx is initial value (Kielhorn et al. the application of ≤10 μl cm−2 of a liquid or 1–5 mg 3.1. For infinite 3. which is derived from Fick’s first law of diffusion meant into and permeation through the skin and the differential mass balance.36 W. a ‘significant reduction’ usually d x occurs when the permeant concentration in the where J is the flux of the permeant (mass per donor phase declines by more than 10 % of the cm2). over time (Crank 1975).g. Depending Kinetically. typically shorter for time) are obtained by evaluating the time-­ lipophilic non-bound molecules. permeation profile of cumulative amount perme- ated across a unit area of membrane versus time. hydrogen bond acceptor or process. nor is it metabolised. the flux profile becomes essen- .1 Infinite-Dose Permeation dose. Lau and K.1 Permeation Kinetics cm−2 of a solid formulation should be used for finite dose studies (OECD 2004a). or due to evaporation. binding sites become An important parameter is the flux. hydrophilic molecules). molecules penetrate into and diffuse through the stratum corneum. It is also assumed that the skin is a pseu. It is also assumed that tion and procedure of the protocols. Generally. permeant con.1. depend on the diffusion coefficient does not vary with the the study aims. Thus.and a steady-state concentration gradient of the ing to Fick’s first law of diffusion (Eq.2.M. the concentration gradient. 3. Initially.2). In J = -D (3. exponentially due to binding and the increasing Typically. and that permeant formulation may be applied in order the barrier properties of the skin remain constant to mimic in vivo application of an ointment. The amount and con. due to penetration of the per. longer for dependency of its cumulative permeated amount. lipophilicity. dt dx centration of permeant formulation to be This equation assumes that the permeant does not applied to the skin surface.2. fusion affects the permeant concentration with centration in the formulation changes during time. Fick’s second law of diffusion (Eq.

 Graph B shows an alternate situation dm where a plateau is reached following prolonged incubation. it is very difficult to mea. Jss can also be esti- h2 mated by L= (3.7  L. m axis. The linear portion of the graph rep. 3.4 into Eq. ation data: resents the steady-state flux (Jss) and can be DK m Cv determined by simple linear regression of the lin. and the curve approaches a ­ athematical model in examining skin perme- m straight line. which is the most widely applied ability coefficient Kp are obtained from an in vitro . L. J ss = (3. conditions. first layer of the skin and h is the membrane Permeant transport across the skin is some- thickness.1  Graph A (solid line) represents a typical A permeation profile for an infinite dose regimen. Since C0 and Cv are related cm/h): to the partition coefficient between donor and K m D J ss membrane (Km). can be where the receptor compartment is at zero determined mathematically by concentration throughout.3  Finite and Infinite Dosing 37 Fig.5) ear portion of the graph (Crank 1975. The intercept on the horizontal Cumulative amount per unit area. essentially a measure for the meant in the donor vehicle.  3. the steady-state flux Jss and the perme- Eq. indicating dt deviation from infinite dose conditions.6) 6D DC0 J ss = (3. t tially constant. from t = t1.4) then substitution of Eq.3 gives Typically. The lag time can be obtained by extrapolating Typically.7) h Cv C0 = K m Cv (3. However. Cv. the concentration of the per. with ‘perfect’ sink conditions Crank (1975) showed that the lag time. The steady-state flux (Jss) is obtained from the B gradient (dm/dt) of the linear portion of the graph. is easily obtainable speed of transport across a membrane (often as or is usually known. 3. Scheuplein h and Blank 1971). coefficient (Kp).3) It has been estimated that the time required for h most permeants to achieve steady-state flux is where C0 is the concentration in the outermost about 2. Practically.5. L. the steady-state flux is assessed the steady-state (linear) portion of the permeation from an in vitro experiment under infinite dose graph to the intercept on the horizontal axis. 3. where Kp = = (3. due possibly to donor depletion or non-sink conditions L t1 Time. times described in terms of the permeability sure C0. where t1 ≥ 2. gives the lag time.7 times the lag time (Barry 1983).

step in skin absorption and may have a significant ity coefficients should only be derived from data effect on the total flux measured. 3. particular caution has 2012. perme- (Franz et al. steady-state flux. infinite the limiting factor (Cross et al. reported (Chen et al. 1993. The most widely used receptor fluid parameters such as D and Km can be problematic is isotonic buffered saline. However. 3. beyond which the amount .4. especially for permanents that have very utilise an infinite dose regimen to define a perme- low fluxes or present difficulties for detection ant’s properties. where a relatively geneous membrane and that permeation through small amount (i. or the diffusional highly lipophilic compounds. then Kp can be determined. This assumption is usually state permeation seldom occurs and.1. throughout the experiment in order to ensure In contrast to the infinite-dose permeation drug permeation is not affected by solubility in profile. It has bovine serum albumin) (Skelly et al.5. valid for most permeants.M. 20 % oleyl ether or 5 % meability coefficients (Shah et al. 2006). 2005. receptor fluid may become the rate-determining Estimates of steady-state flux and permeabil.7 are commonly limitation of infinite dosing is its failure to mimic used to evaluate infinite dose experiments. 6 % state is established leads to false estimates of per. if the cells. 2011). Skelly ation of permeants. been recommended that infinite dose permeation OECD 2004a).38 W. it is the application of topical drug formulations in assumed that the skin membrane acts as a homo. for as the membrane thickness (h).g. steady-­ transdermal transport. Typically. polyethylene glycol. highly lipophilic or highly hydrophilic mined. 1994). even under infinite dose conditions. a major Although Eqs. the dose conditions may result when finite doses are equations assume perfect sink conditions applied repeatedly. When solubility points beyond 2. therefore. to be exercised in experiments where prolonged although shorter durations have also been incubation times are necessary. as calculations for other et al.2. pH 7. although in some cases. 2001). the solubility in the path length. an increase in exposure time depletion of the donor phase. is often unknown. 2010. These factors may result in inaccura- experiments are performed for 24–48 h (Boonen cies in steady-state flux and lag time estimations et al. ability coefficient and lag time. 1987). using data before steady rier or a solubilising agent (e.7 times the lag time when is a concern. Jss but this may lead to dilution of the penetrant is obtained from the gradient of the linear portion below the detection limit. For static diffusion of the permeation profile. Howes et al. However. 1997).W. As described above.2 Finite-Dose Permeation cases for an infinite dose to establish steady-state In vitro percutaneous absorption studies often flux. finite-dose application may result in a the receptor phase. flow-­ ‘pseudo steady-state’ condition. and therefore. non-sink receptor may alter the integrity of the skin barrier conditions and deterioration of the skin can occur (Kleszczyński and Fischer 2012). it is commonly regarded that sink condi- concentration of the permeant in the applied tions are maintained when the receptor fluid does vehicle (Cv) is known. 1987.e. where the through diffusion cells maintain sink conditions amount permeated may be transiently linear but by constant replenishment of the receptor fluid. then reaches a plateau. 1996). Most in vivo experiments are based on molecules. but for highly viscous the permeability coefficient cannot be deter- vehicles. the receptor phase can include lipo- (pseudo-) steady-state conditions are established philic solvents which do not affect the skin bar- (Kielhorn et al. However. 2012. Karadzovska et al. Brain et al. Under these circumstances. with time. experiments should last for at least 24 h (OECD In reality. Also. 2004a). centration of the penetrant (Ng et al.6 and 3. that is. partitioning behaviour may become finite dosing. a finite dose) of the formula- the stratum corneum is the rate-limiting factor for tion is used. (Moody 2000). Walters et al. Therefore. Fasano et al. Lau and K. Ng infinite dose experiment. not contain more than 10 % of the saturated con- Kp is often used to characterise the skin perme. It has been suggested that 72 h or even longer may be needed in some 3. 2012. 50 % ethanol. common clinical situations. For in vitro studies.

since δ is considerably smaller in compari- permeation area) versus time. are known. C0 is Tmax the concentration of the permeant in the first Time. leading to maximum flux is referred to as Tmax h2 Tmax = (3. A Depletion of the donor Cumulative amount per per unit area.3 and 3. Wilkinson et al.4). The maximum flux (Jmax) tal problems can ensue.2). by plotting the amount pene. D is the apparent diffusion coefficient. 2006). Given that steady-state flux may not be obtained before B dm donor depletion becomes significant.8) h2 h2 − d 2 Tmax = (3. instantaneous Jmax and Tmax are sometimes obtained graphically flux) against time (Figs.9) 6D Here. the maximum flux may not represent steady-state flux dt Time. These can be represented by (Scheuplein and Ross 1974. 3. the vertical axis represents instantaneous flux. h is the thickness of the stratum corneum and δ is the thickness of the Fig. For a finite of amount penetrated per unit time (assuming constant dose. 3  Finite and Infinite Dosing 39 Fig. δ can be neglected. since applying a small . Kasting 2001) 1. 3. Thus. 3. experimen- 1983. 3. it is possible to estimate the apparent diffu- tion (Fig. 2004.3  Estimation of maximum flux (Jmax) from a graph finite dose layer on the skin surface. The maximum flux (Jmax) is obtained from the gradient (dm/dt) of the near-linear portion of the curve in region B.2 and 3. trated between the time points (i. In this case. t layer of the stratum corneum. Kasting 2001). 3. sion coefficient (D). In addition to this mathematical model.e. Crank 1975. t and the time to maximum flux (Tmax) are the most Amount penetrated per unit time Jmax commonly reported parameters in finite dosing.85 DC0d J max = (3. The time taken to reach son with h.3) (van de observed which corresponds to the maximum Sandt et al. flux before appreciable donor depletion (Franz Due to the nature of finite dosing.10) 6D permeated remains constant due to donor deple. if the values of Jmax and Tmax Alternatively. m phase gives the character- istic plateau in region A. a peak is from experimental data (Figs.2  A typical permeation profile for a finite-dose regimen.

Youenang Piemi 2008). 1999) of the remaining drug from and Zatz 1986). 2003. homogenous distribution (Kasting and Miller brane in vitro can be difficult. Since the 2006. However. Figure copyright (2012) reprinted with permission finite doses of loperamide and propylene glycol (PG) from Elsevier) through human skin over time (Data are from Trottet et al. 1998) to help distribute the formulation assumed in many reports. Some researchers recommend the application device. Zhai and Maibach 2001. affecting the integrity of the skin mem- tion over the whole donor area is needed to mini. 40 mg/cm2 PG 10 mg/cm2 300 Loperamide flux (ng/cm2/hr) 1000 PG 40 mg/cm2 PG Flux (µg/cm2/hr) 800 200 600 400 100 200 0 0 0 4 8 12 16 20 24 Time (hours) Fig. Brain tions have investigated drug formulation distribu- et al. 40 W. the volatile solvent applied per unit area (Wester and Maibach 1976. simulate the in-use conditions of topically applied 2007).4  Exemplar data showing instantaneous flux of (2004). homogenous distribution is generally et al. or by weight difference that the skin be exposed to ambient conditions to (Walters et al. a very limited number of publica- (Hadgraft et al. Franz et al. 2007. volatile medium to allow easy application and Grégoire et al. influence the absorption rate in some cases ing. Sometimes. but this mechanical stress may nously poses more complications than liquid dos. drug permeation (Akhter and to the skin. Southwell and Barry 1984). 1993. However. 1993). leaving a thin film of the drug finite doses depends on the amount of drug on the skin surface. Gupta et al. Vallet et al. ing experiments can affect skin hydration and. Ng 1400 400 Lop. evenly on the skin surface. may extract some lipids from the stratum cor- Franz et al. The volatile ­bioavailability of drugs applied to the skin in solvent evaporates. Another method used for uniform dose mise variation within and between experiments. ysed in order to determine the actual dose applied consequently. To date. brane. Trottet et al. In this case. 1995. Lau and K. 1999) or spatula (Dreher tion over the incubation area (Lademann et al. 10 mg/cm2 1200 Lop. 2009). Semisolids are usually applied manually. 1994.M. it is The degree of occlusion of diffusion cells dur- important that the application device is also anal. a homogenous drug distribu. 2002. 1997. Lademann et al. the permeant is dissolved in a formulations (Wilkinson and Williams 2002. finite dose formulation evenly across a mem. using a mechanical device such as a glass rod 2009). distribution is by massaging the formulation onto Applying semisolid formulations homoge. evaporation of . This is commonly done by extraction Barry 1985. 1995. 3. Sarpotdar (Gupta et al. the skin surface. 2004. et al.W. neum. Brain et al. (Amidouche et al.

Walters et al.2.2. Maibach 2002). Selzer et al. 1995. Thomas and Heard 2007. most commonly used to assess the change in per- for example. For example. then extracted and determined to calculate the lines state that the mean recovery of the permeant diffusivity and solubility of the drug within the and metabolites should be in the range of stratum corneum. and its bar. but may be 3. phase. second law of diffusion (Anissimov et al. The OECD guide. recovery can be corneum.Dn 2p 2 t ö ù (3. b). be performed over 7 days (Williams 2003). 2012). extraction methods. 2001. it is important that a Dreher et al. Occlusion is also used during finite dosing be achieved. or to pre. This is a method. Müller et al. 2012. 1998). 1999. 2013): éæ x ö 2 ¥ 1 æ np x ö æ . 2010. t ) = KC0 êç1 . care is required to degrade permeants in situ (Lau et al. European Commission 2006). (OECD 2004a). 2005. if the can be valuable. contact with the test substance (e. 2011). Okuda et al. 1998). under cer. skin esterase has been shown to Koyama et al. In some rier function can thus be affected (Selzer et al. constructing a drug concentration/depth profile ues for at least 24 h. the test of a volatile substance function of position and time within the stratum that had to be trapped in a filter. failure to detect of permeant absorption over a realistic period of degradation products using suitable analytical time that relates to potential human exposure methods may confound experimental results. the donor and Moreover. the skin may become dry. In such cases.11) c( x . Degradation of the permeant vent evaporation. whereby mass balance is performed on completion of the the stratum corneum is progressively removed by experiment to ensure the total amount of applied adhesive tape and the drug within each tape is permeant could be recovered.3  Finite and Infinite Dosing 41 the vehicle and/or permeant can occur in non-­ should include permeant remaining in the donor occluded experiments which can impact signifi. Exposure times of up to 48 h are commonly used (Hadgraft et al. 2007). the recommended total recovery may not 2013). 2011). it is possible to predict not met (OECD 2004a. used in finite dose studies. On the other hand. avoid damage to skin integrity caused by exces.2.å sin ç ÷ exp ç ÷ú ëè h ø p n =1 n è h ø è h2 øû . 1995. the experiment design should mimic meant concentration throughout the stratum cor- the application exposure period and thus should neum (Lau et al. Due to the limited amount of test substance Mohammed et al. 2004. for example. that within the skin and all equipment in cantly permeant absorption (Frasch et al. data collection commonly contin.g. 1996. Gupta et al. 1997.÷ . exposure times usually ucts are likely to have lower permeation rates reflect ‘in-use’ scenarios to allow quantification than the test compound. However. port parameters across skin to be determined. or in 3. 1994). The mass balance Moser et al. receptor chambers of the diffusion cell). and reason must be sought if this is Under infinite dosing.2 Penetration Kinetics shorter to mimic exposure to rinse-off products (Brain et al. 2003). 100 ± 10 %. Thus. Moody et al. However. 2012. 2010.1 Infinite-Dose Penetration studies on occupational exposure to hazardous Although skin permeation data allow the trans- materials (Howes et al. which could lead to a change in could also aggravate the accuracy of total recov- permeant concentration (van de Sandt et al. analytical detection or difficulties with simple tion of small amounts of formulation. 1999. 2003. due to the limits of to avoid drying of the skin surface due to applica. Stinchcomb et al. cases. t)) as a tain circumstances. using the appropriate solution of Fick’s broadened to 100 ± 20 % (OECD 2004a. the concentration of the penetrant (c(x. Pellett et al. The degradation prod- For finite dose studies. Chemical degradation could also have a signifi- sive stratum corneum hydration (Zhai and cant impact on permeant stability (Kubota et al. The tape-stripping technique is drug formulation is for a once-a-week product. ery. Weerheim and Ponec 2001.

New York Boonen J. Dressler WE. If the concentration of the permeant in the stra- tum corneum is obtained experimentally. It is also assumed that the neum (where 0 ≤ x ≤ h). characterise and compare the skin through human skin in vitro: application from cos- absorption kinetics relating to finite or infinite metic vehicles. with finite dosing. Poelman M-C. Ng In Eq. Oxford Cross SE.42 W. K is the partition coefficient between diffusion. permeant concentration is to be determined and and the stratum corneum is the rate-limiting D/h2 gives the characteristic diffusion parameter. Lau and K. Benson HAE. Oxford References University Press. 3. Barry B (1983) Dermatological formulations: percutane- ous absorption. James VJ. Liu X.11. Again. Malysheva SV. the test permeant.12) n =1 b + b 2 + a n2  h   where Cv0 is the initial concentration of the per. the formulation should not the stratum corneum and the formulation vehicle. Kelling CK. Mudumba S.12 (Kasting 2001.2.M. hv cyclodextrin complexation. it is possible to into Eq. Montassier P. β = K(h/hv). Adv Drug αn∙tan αn = β. Dressler WE (2005) Percutaneous pene- tool for transdermal delivery.2.t ) = 2 KCv 0 ∑ exp − 2  (3. 2013): ∞ b cos (a n x / h ) − a n sin (a n x / h )  a n 2 Dt  c( x . Acta Derm Venereol 78:186–189 . Taevernier L. alter the membrane integrity or act as a carrier for x is the vertical position within the stratum cor. attenuation of tretinoin induced skin irritation by Î2-­ meant in the donor compartment. Int J Pharm 408:223–234 Crank J (1975) The mathematics of diffusion. Toxicology ent applications in transdermal delivery. Food Chem appreciate the underlying assumptions and Toxicol 43:681–690 Chen M. t is the time at which the experiment is maintained under sink conditions. occlusion and the penetra.2 Finite-Dose Penetration experimental concentration profiles can be fitted Similarly. dosing. In this respect. Moloney SJ. barrier. In addition. De Spiegeleer B (2012) Human skin Finite and infinite dose regimens have differ. J Pharm Dreher F. Fahr A (2011) Skin penetration and deposi- limitations of the models. Dancik Y. Diana di Mavungu Conclusion J. Ademola J. Mathematical models allow us to Percutaneous penetration of dimethylnitrosamine predict. Walters KA. deposited drug films.11 to determine K and D/h2 (Pirot et al. Int J Pharm 111:111–116 Anissimov YG. However. tion of carboxyfluorescein and temoporfin from differ- ent lipid vesicular systems: in vitro study with finite and infinite dosage application. they are an invaluable Sharma RK. it is important to application from cosmetic vehicles. Food Chem Toxicol 33:315–322 Brain KR. Each 301:21–32 Brain KR. Green DM. oxybenzone? J Invest Dermatol 117:147–150 tion enhancer N-methyl-2-pyrrolidone. Loretz LJ. However. Jepps OG. Duchene D ing human Stratum corneum removed by adhesive-­ (1994) Evaluation by laser Doppler velocimetry of the tape stripping. Jiang R. Hostynek JJ. 3. Walters KA. predict permeant concentration at a given posi- 1997). Arens A. Pharmacol 37:27–37 Maibach HI (1998) Colorimetric method for quantify- Amidouche D. Marcel Dekker. Brain S. Roberts MS (2001) Can Akhter SA. using Eq. reduce the human skin penetration of the sunscreen ation. De Saeger S.W. Selzer transports across the stratum corneum by passive et al. 3. Roberts MS (2012) being the theoretical height of the volume of Mathematical and pharmacokinetic modelling of epi- donor formulation and αn is related to β such that dermal and dermal transport processes. Gettings SD (1995) challenges. penetration of selected model mycotoxins. in tration of diethanolamine through human skin in vitro: applying these models. very few studies have Deliv Rev 65(2):169–190 made use of this mathematical model. effect of dose vari. Barry BW (1985) Absorption through human increasing the viscosity of formulations be used to skin of ibuprofen and flurbiprofen. the 3. Howes approach presents its own advantages and D. it is assumed that the skin is a tion inside the stratum corneum and at a given homogenous membrane and that the permeant time.

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van de Sandt the workshop on principles and practices of in vitro J. Paris loperamide hydrochloride. Shah VP. Greaves L. Toxicol In Vitro Negligible penetration of incidental amounts of alpha-­ 21:1182–1190 hydroxy acid from rinse-off personal care products in van de Sandt JJM. Barry BW (1984) Penetration enhancement Williams A (2003) Transdermal and topical drug delivery in human skin.M. Maas W. Regul Toxicol Pharmacol Guy RH (1997) Characterization of the permeability 39:271–281 barrier of human skin in vivo. Guy RH. Percutaneous absorption of solvent corneum lipid profile by tape stripping in combination deposited solids. Zatz JL (1986) Evaluation of penetration (1997) Percutaneous penetration of octyl salicylate from enhancement of lidocaine by nonionic surfactants representative sunscreen formulations through human through hairless mouse skin in vitro. solutions evaluated with an in vitro stratum corneum Williams FM (2004) In vitro predictions of skin absorp- tape stripping technique. Abdel-Mottaleb MMA. Fuchs A (2011) thickness pig and human skin. J Invest Dermatol 67:518–520 Shah JC. Touraille GD. Payan JP. Kaufman LE. simple emulsions. Int Arch Occup Environ 256–257 Health 79:405–413 Southwell D. 28. Montomoli L. Donahue DA. Schaefer UF. Hadgraft J. physicochemical properties of test compounds in per- availability and bioequivalence. skin permeation evaluation: the only realistic option. through hairless rat skin in vitro: effect of multiple and neum in vivo from volatile and non-volatile solvents. Sartorelli P. evaluations and predictions. Yacobi A (1987) FDA and AAPS report of Wilkinson S. no. Stinchcomb AL. Arch Dermatol Res 293:191–199 Neumann D (2013) Finite and infinite dosing: difficul. Int J Pharm 22:291–298 Youenang Piemi MP. Watkinson AC. Dick I. Avalos J. Pirot F. OECD. Kaka I. Food Chem Toxicol 35:1219–1225 75:176–181 Walters KA. mamide and increased hydration on finite dose perme. Simion phosphorus compounds using full-thickness and split-­ FA. Williams F (2006) Interactions of skin thickness and percutaneous penetration studies: relevance to bio. skin absorption: in vitro method. Int Arch Occup Environ Health Skelly JP. Limasset model. Physiol Rev 51:702–747 Int J Cosmet Sci 20:307–316 Scheuplein RJ. J Invest Dermatol 62:353–360 with high-performance thin-layer chromatography. Maibach HI (2001) Effects of skin occlusion on Thomas CP. Williams F (2002) Effects of experimental Analysis of percutaneous permeation data: II. Int J Pharm 171:207–215 Pharm Res 16:1288–1293 Zhai H. Howes D. Toulon M. Cage S. effect of 2-pyrrolidone. J Pharm Sci skin in vitro. Blank IH (1971) Permeability of the skin. Heard CM (2007) Probing the skin perme. Cruz C. Wester RC. Int J Pharm 151:91–98 tion of caffeine. Hahn T. Kraus AL. Appl Skin Physiol 14:1–10 Comparative depth profiling and metabolism of eicos. 94:1562–1567 Teetsel NM. Story DC. Nielsen JB. Maas WJM. Skin Pharmacol ation of eicosapentaenoic acid and ketoprofen 2. Proc Natl Acad Sci Walters KA. Nielsen J. Lau and K. de Luca M. Keating G. Pharm Res 4: cutaneous penetration studies. Tenjarla S. Bunge AL. Ponec M (2001) Determination of stratum ous absorption. skin barrier apentaenoic acid. percutaneous absorption: an overview. Roberts MS. 75:519–527 Flynn G. centre comparison study. Kenyon S. Fautz R.W. Maibach HI. Pellett MA. Vallet V. Schaller KH. Brain KR (1998) In vitro Scheuplein RJ. Hadgraft J (1997) Supersaturated Robinson E. Seiller Stinchcomb AL. Brain KR. Chow D (1994) Wilkinson S. OECD.44 W. dose and percutaneous absorption in rhesus monkey Adv Drug Deliv Rev 65(2):278–294 and man. Toxicol In Vitro 25:2041–2047 JC. Bazire A. Ross LW (1974) Mechanism of percutane. Josse D. dimethylfor. Int J Pharm 274:213–219 OECD (2004b) OECD guideline for the testing of chemi. testosterone. Lau SWJ. van Burgsteden JA. Gettings SD Sarpotdar PP. James VJ. and benzoic acid: a multi- Pirot F. Carmichael human skin using an in vitro static diffusion cell PL. Lallement G. Skin Res Technol 8:1–6 Trottet L. Boudry cals 428. Sakaguchi H. Mirza M. Maibach HI (1976) Relationship of topical ties in measurements. Int J Pharm 109:283–290 human skin in vitro. Korinth G. Pharmaceutical Press. Eur J Pharm Biopharm 67:156–165 function. Kalia YN. Wester RC. from theory to clinical practice. Bunge A. Wilkinson SC. Maibach HI (2002) Occlusion vs. Selzer D. Paris I (2007) In vitro percutaneous penetration of organo- Okuda M. Weerheim A. Zhai H. Guy M. V. London ation of aspirin and caffeine. Merly C. Larese F. Evaluation conditions on absorption of glycol ethers through of the lag time method. Marty J-P (1998) Transdermal delivery of glucose RH (1999) Chemical uptake into human stratum cor. Grossiord J-L. Watkinson AC. Ng OECD (2004a) Guidance document for the conduct of e­ nhancement of in vitro percutaneous permeation of skin absorption studies. Davis AF (2004) Effect of finite doses of propylene glycol on .

... Ryan • I............ Adelaide..... 49 4....... Moghimi.......................11....... 56 References...... 64 4......E......3 Structure-Permeation Relationship 4......... Adelaide..S........edu............................ Haridass • Y..... Therapeutics Research Centre. Australia GPO Box 2471............. Zhang Therapeutics Research Centre...... School of Medicine............7 Skin Hydration and Percutaneous Conclusion..R. Isha Haridass..................... School of Pharmacy University of Queensland...12........ Roberts Contents 4.... School of Pharmacy University of Queensland........ SA 5000............ Moghimi Therapeutics Research Centre.................. Adelaide.. Institute.. Institute.... 67 That Favour Appendageal Permeation................ Maibach (eds.11 Barrier Performance in Cosmetically 4....8 Age-Related Skin Barrier Performance.2 Chemical Depilatories.....................3 Peeling Agents.......... Australia and Medical Sciences..... Woolloongabba............ 66 4............ 65 4.... Australia QLD 4102....... University of South Australia................... School of Pharmacy SA 5000... 60 4.... School of Medicine..1 Introduction.... Australia of Medical Sciences.. QLD 4102........au © Springer-Verlag Berlin Heidelberg 2017 45 N. Mohammed Q... Translational Research and Medical Sciences... 48 4. Iran e-mail: m.................1007/978-3-662-53270-6_4 .................... Hamid R.... Elizabeth Ryan.5 Physicochemical Properties 4..........11........12. 69 Absorption. 65 to Targets for Drugs and Cosmetics........ 68 4.9 Site-Related Skin Barrier Performance..... Dragicevic......12 Compromised Barrier Performance.....2  Thermally Damaged Skin and Its 4....... University of South Australia........ Tehran. University of South Australia...... 46 4.13 Prevention of Percutaneous Permeation: and Deeper Tissue Permeation..... 66 in Percutaneous Absorption.11... Qian Zhang..... Australia M....................10 Gender-Related Skin Barrier Performance........ Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin......4 Other Cosmetic Products.. SA 5000............. 67 Favour Direct Subcutaneous 4.)........... DOI 10..... 67 4............. Yousuf Mohammed.1 Insect Repellents..........6 Physicochemical Properties That Manipulation...... Shahid Beheshti University GPO Box 2471..4 Permeation Through Intact Epidermis.. Therapeutics Research Centre. Grice........... Grice • E.. Non-formulation Parameters That Affect Penetrant-Skin-Vehicle 4 Interactions and Percutaneous Absorption Jeffrey E...... 56 A Possibility and a Necessity. 62 J... 46 4.... H.....roberts@uq......1  Intrinsic Barrier Defects..... 69 4... School of Pharmacy....... Translational Research and Medical Sciences..... GPO Box 2471... and Michael S.. 53 4...... Woolloongabba.....2 Skin Responsiveness as It Applies Treated Skin.. Therapeutics Research Centre. Roberts (*) H..I... 67 4.......11................

gel. the regional applica- tion arises not only from biological differences tion of drugs aims to target areas beneath the site in skin composition between sites on the body. minimal systemic lation components that these interact with) and absorption is desirable. In addi. ceous glands (anti-acne drugs). (Fig.g. the effects of hydration ageing) and for local protection (such as the and related conditions on percutaneous absorp- insect repellent.1). some induced alteration of skin barrier and various may be irritants and others toxicants.N-diethyl-meta-toluamide tion. skin-­penetrant-­vehicle interaction and percuta- tion. minimal systemic absorption is conditions (e. permeation through thermally damaged (DEET) and barrier creams).46 J. anti-inflammatory creams) and meation through intact stratum corneum (SC) cosmetic purposes (such as to slow down skin and skin appendages. advancement of skin science has resulted in an in-depth characterisation of the skin barrier per- formance and an understanding of the different 4. In this type of drug delivery. Skin targets for local by modifying skin permeability through the use delivery include the skin surface (sunscreens. There are Applies to Targets for Drugs two separate needs that a topical product fulfils: and Cosmetics its cosmetic properties. cosmetic- these products are relatively innocuous. dry skin or psoriasis) but is also desirable. 4. burn injuries and some pen- The skin is a site of non-invasive delivery for etrant physicochemical properties that can affect drugs. we discuss in this chapter the as from the deliberate use of topical products for required molecular properties for optimum per- therapeutic (e. is reflected in a range of skin per. are the targets. lotion. the skin is exposed to a wide variety of neous absorption. agents) and the epidermis (anti-inflammatory meabilities and skin responsiveness. be optimised for higher percutaneous absorption powder. nicotine for minimal penetration of an excipient through the smoking cessation and nitroglycerin for angina skin. These can include the use of different types of the delivery system and skin conditions should formulations (paste. The varia. of application.g. cosmetics and diagnostic agents. cream. . and minimal skin retention.).1). skin varies enormously in its biol. e. where the effect is lim- manipulating formulation (vehicle) properties ited to the skin itself and different parts of skin (such as pH. manipulating the thermody. 5) and the selective delivery of temic delivery or regional effects. delivery. In gender. etc. sys- effects (see Chap. In contrast to this is namic activity of the active and excipients by local or dermal delivery. The biological variables. drugs) (Fig. (depilatories or hair growth stimulators).1). hair follicles enhancers. patch.g. In systemic the active ingredient for a local or systemic ther. 4. between species.1 Introduction temperature and humidity). Finally. as it applies to the stratum corneum (hydratants and keratolytic this chapter. ointment.g. skin variability can greatly impact on the rate ronment (such as exposure to pesticides and and extent of skin permeation for both active allergens). age or the presence of particular skin such cases. the prior application of topical products. a range of strategies can be employed. a drug is targeted to remote areas apeutic effect. N. seba- However. although not always achievable determined by environmental conditions (e. as well and excipients. sweat ducts and ogy between and within individuals as well as sweat glands (antiperspirants and deodorants). including its sensorial feel arising from its emollient and hydrating Drugs are applied to skin for local action. active concentration and the formu. In order to achieve the latter with through systemic circulation. such as joints and muscles. etc. 4. As these various sources of unwanted substances each day – from the envi. the workplace and the home. cleaning agents). of occlusion and the presence of penetration insect repellents. In this case. 4.E. This variability.2  kin Responsiveness as It S factors that can impact on this barrier. pectoris (Fig. skin absorption retardation. Whilst most of skin. Grice et al.

but unwanted contact of the skin with aminopeptidases and peptidases. and for Phase 2 (conjugation) are 0. the activity of cutaneous of metabolising enzymes and transporters in the CYP1A1. associated with the receptor populations in the various skin types. The importance of location is illus- porters in the skin (Dancik et al. a member of the Cytochrome P450 various skin regions and the variation in target superfamily of enzymes. A (Hotchkiss 1998) suggests that the activities of good understanding of the skin barrier and the cutaneous metabolising enzymes for Phase 1 conditions that affect its barrier performance is metabolism (oxidation. dermis. liver. As metabolism of benzo(a)pyrene and phenanthrene. pig > human. variations in skin permeability include the actions As one illustration.1  Schematic illustration of skin (showing epidermis. cos. cosmetic or diagnostic epoxide hydrolase. is in the order SENCAR mouse > hairless guinea tion of various metabolising enzymes and trans.6–50 % of those for the same enzymes in the minimising any unwanted effects. benzo(a)pyrene hydroxylase (BPH). hypodermis and skin appendages) and underlying muscles and joints. regions for therapeutic. many of trated in the work of Coomes et al.1–28 % and metic and diagnostic delivery systems and for 0. esterases. She also summarises the species and location Sources for skin responsiveness in addition to variations for the various metabolising enzymes. 2010). Hotchkiss toxic and hazardous material can also occur. 4. reduction and hydrolysis) essential for the development of therapeutic.4  Non-formulation Parameters Affecting Skin Permeation 47 Surface: Hair: Insect repellents Depilatory agents Sunscreens Conditioners Antimicrobial agents Colors Stratum corneum: Moisturizers Keratolytics Epidermis Viable epidermis: Skin lighteners Anti-inflammatory agents Sebaceous gland: Anti acne products Dermis sweet gland Dermis: Anti-inflammatory agents Antihistamines Anaesthetics Sweat glands: Antiperspirants Hypodermis Systemic delivery: Nerves Nitro glycerin Blood vessels Lymph vessels Nicotine Muscles and joints Regional therapy: NSAIDs Fig. . Cytochrome P450. showing targets of drugs and cosmetics applied to skin for local. purposes. 7-ethoxycoumarin O-deethylase. shown in our recent review of the range and loca. systemic or regional action Chemicals are targeted to the aforementioned hydroxylase. (1983). in the enzymes have either hydrolysis or oxidative which it is shown that for 7-ethoxycoumarin type activities and include aryl hydrocarbon O-deethylase.

skin is not a homogenous that was about 25 % of the in vivo activity.5.1 (Dancik et al. 1984). related to that in vitro by Eq. while Lockley transdermal ointment (Nakashima et al. 2. bioavailability for nitroglycerin in rhesus mon- ceous and basal cells.glucuronosyltransferase The effect of the metabolic activity on epider- (UGT) and glutathione S-transferase (GST).0.8 % (Wester et al. Wester et al. while both freezing and heat separation led drug candidate should possess for transdermal to a 40–50 % reduction in esterase hydrolysis of delivery are a low molecular size. is also an inhibitor of its cutane- ous metabolism to estrone in the viable epider. inhib. always plays full-­thickness human skin at 60 °C for 2 min to an important role in its passive delivery through separate the epidermis resulted in a 35 % reduc. oxidation of aldehydes from ethanol and glycol 2008): ether. in particu- for 24 h (Bickers and Kappas 1978). the loss of nitroglyc- to glycol ethers can lead to skin sensitisation and erin is due to retention in the skin and tissue bind- irritancy as well as systemic toxicity as a result ing (Imhof et al. a long-standing dermatologic treat- ment. in healthy humans. membranes had methyl salicylate esterase activity However. the fraction of solute enter- activity of all enzymes may be induced. et al. the mal flux can be expressed as a first pass effect. for 24 h led to a two. 2005). 1984). Based on our knowledge of transdermal tion in phosphatase activity responsible for the penetration and skin structure. reported variable induction of cuta. 4. relative sebaceous cell to epidermal basal activities whereby the flux of a solute entering the body were: 6.1) to complete inhibition. i. with much lesser effects on keys to be 56. the treatment Relationship of psoriasis and ectopic dermatitis patients’ skin in Percutaneous Absorption with coal tar. Coomes et al. 1984) and 68–76 % from a corticosteroids (Finnen et al.9 and 3. 1991). a low melting point and relatively skin (Lau et al. the steady state skin flux concentrations (5 μM) of disulfiram inhibited skin Js (= amount penetrated/time period) in vivo is cytosolic aldehyde dehydrogenase 1 (ALDH1). a moderate acetylsalicylic acid (Aspirin®. Bayer) in porcine lipophilicity. after topical application is modified by the skin Importantly. (1998) found few hydrogen bonding groups. 1983). from Nitroderm® TTS patches manufactured by neous 7-ethoxycoumarin O-deethylase by various Novartis (Imhof et al. in vivo = F × J ss .6 ± 5. Similar AHH induction was seen in excised drug candidates for application via this route are normal human breast skin incubated with coal tar very few.e. Low activity in in vitro skin. as we know. it is generally hydrolysis of diisopropyl fluorophosphate (Loden accepted that the optimal molecular properties a 1985). the ity. whereas high concentrations (500 μM) led J ss .48 J. have pointed out that the dermal exposure As well as skin metabolism. The nature of the penetrant. Ethanol. in vitro (4.E. intact solute concentration-time profiles (AUC) also showed that topical treatment of the skin with after topical and intravenous dosing: F = AUC beta-naphthoflavone induced 7-­ ethoxycoumarin (topical) / AUC (iv). ery have been recognised for a long time. as reviewed by Hotchkiss. Such require- that both full-thickness and separated epidermal ments are discussed in more detail below. For instance. the biovailability (F).2. 4. the skin. of their oxidation by cytosolic alcohol and alde. In the event of the complete loss of metabolic hyde dehydrogenase (Lockley et al. 1983). 2012). Heating of lar its physicochemical properties. 3. uridine 5′-diphospho. Cross et al. a penetration enhancer for the transport of beta-estradiol across the epidermis.3 Structure-Permeation mis (Liu et al.to fivefold induction Even though the advantages of transdermal deliv- of aryl hydrocarbon hydroxylase (AHH) activ. 1987). barrier and contains a complex epidermal path- . ing the body intact after topical application. In addition. it was estimated to be 75 % Finnen et al. Grice et al. while the other enzymes studied (Coomes et al. cal- ited or destroyed by various agents and processes culated as the ratio of area under the plasma (Hotchkiss 1998). found the % O-deethylase by more than tenfold for both seba. respectively.

1. dermal perfusion also affects and it is difficult to find a general relationship to the clearance of solutes from the dermis and the describe skin permeation overall. the viable epider. Roberts et al. 4. Even the stratum corneum (SC) contains parallel and serial path. (Fig. the intercellular and transcellular routes. This trans. through hair follicles J sskin = ç sc + ve + dermis ÷ + J sapp (4. 4. viable epidermis. mal concentrations. 1975. aqueous diffusion layer and dermis may form. geal pathway will be discussed in relation to Skin is a complex barrier. sc. therefore. which. but they [modified from (Zhang et al. Two main flux (Js) through the skin and is defined by the approaches have been used in defining these amount of solute (Q) passing through an area of relationships: one is to express relationships in skin (A) over a period of time (T) at steady state terms of observed fluxes through the skin and (i. structure-permeation relationship dependency relates to the nature of the barrier As we will see later. melting ute partitioning into and permeation through the point and molecular size (molecular weight or appendageal pathway (App). etc.3) and other skin shunts.4  Non-formulation Parameters Affecting Skin Permeation 49 way. recognising that the flux of solutes from satu- de) and appendageal pathways (app) (Eq. in turn. in its simplest mis. the second appears A ×T to arise from the conventions associated with . Potts and Guy the VE. The two approaches differ in that given vehicle (fv) (Roberts 2013) (Eq. Lien and Tong 1973. In contrast. the first has its origins in the work of Higuchi in principle.3): rated solutions vehicles that have not affected the skin permeation should all be the same Q J sskin = = J max f v (4. water partition coefficients. 1977.2) (Roberts 2013). are different parallel penetration routes via the intact epidermis and the appendages (hair folli- cles and sweat ducts) as well as serial barriers. lish a relationship between skin permeation and The SC is considered to be the main barrier to the molecular structure descriptors of solutes. The dependency of skin penetration on lipid- mal permeation process.e. Considerable efforts have been made to estab- ways. Besides consid. and -1 æ 1 1 1 ö an appendageal pathway. dermis. partitioning into molar volume) has been widely discussed in and permeation through the SC (transcellular or early studies published in this area (Scheuplein intercellular routes). as illustrated in Fig. transdermal delivery in most cases. ve. 4. finally uptake by blood vessels or permeation the SC was considered as the main barrier to through the hypodermis to underlying tissues permeation of drugs through the skin. physi. The transder. T > an apparent lag time for permeation to the other in terms of the physicochemical deter- occur and can be related to the maximum flux minants often used to understand the variability (Jmax) (that will be similar for all vehicles not in fluxes between different solutes and different affecting the skin) and fractional solubility in a formulations. Epidermis mis. partitioning between the SC and Blank 1971. recognised that the underlying viable epider- port is expressed quantitatively. the maximal permeation from a given vehicle. solubility. 2002). Du Plessis et al. may affect viable epider- permeation through the intact epidermis. tum corneum. In è Js Js Js ø addition.4 Permeation Through  Intact including the stratum corneum. In these studies. 4. as the steady state rate of permeation or also contribute to the skin resistance. is related to the individual fluxes for (Higuchi 1960) and often relates permeation to solutes through the individual skin layers (stra.2). In this section. and 1992. 4. involves sol. the dermis. clearance of solutes across the dermis into deeper ering a structure permeability relationship for tissues. 2009). through which there physicochemical properties of molecules. permeation of cochemical properties that favour a follicular drugs through intact epidermis and the appenda- pathway are also considered in this section. Michaels and viable epidermis (VE). dermis and aqueous diffusion layers. which. permeation through et al. residing mainly in the stratum corneum.1).

E.5) neither the formulation nor the solute affects D the membrane. recognising most of the previous this area is the accuracy of the predictions for studies. and a partition coefficient Ksc. the path length for transport across in solution in a supersaturated form. This the effect of diffusivity on skin transport. and solutes could be the SC (Dsc).v) is given by (Ssc/Sv) yields: advantage of directly showing the maximum dose deliverable over a period of time (Magnusson et al. 2004): melting point above 25 °C (Yalkowsky and log J s . Ssc. Ssc = K sc . a changed of the solute in the SC. sat = = sc Ssc Milewski and Stinchcomb 2012). Q D J sskin . the during solute partitioning or the diffusion permeability coefficient is the product of this par- process.1 and and therefore be associated with higher fluxes. 2002): log Ssc = log K sc .688 (n = 278. if A × T hsc (4.0. 4.48 log P .4) Valvani 1980) to yield (Roberts et al. 1987). Relevant to the first approach.77 (4. the diffusivity is a function of solute size.4.012 ( MP . expanding by recognising that the SC-water par- The use of saturated or maximum flux has the tition coefficient (Ksc.52 . it is evident from Eq.50 J. v ´ Dsc / hsc ). tition coefficient multiplied by Dsc/hsc (i. incorporating these into Eq. It does not reflect the con- ited by the variability in permeability coeffi. The most recent combination of Eqs. Sloan et al.4. octanol-water partition coefficient and the solute p < 0. either and vehicle (i. tribution of solubility of solutes to the flux. whereas the permeability coefficient depends on the formulation used (Zhang et al.0141MW (4. v Sv = k pskin Sv hsc thermodynamic description of the penetration process. 1996). However.e.25 ) + 0. 4. as ability coefficient. 4. Grice et al.3 and 4. can be expressed in maximum flux after formulation application terms of a solubility of the solute in the vehicle may suggest an alteration of the skin bioproper. Scheuplein 2013). 2006.001) (Magnusson et al. v ´ Sv ) and. i. (Kasting et al.sat = log J max = . Care should be applied for solute penetration and when the saturated sol- when using such relationships. above the SC (hsc) and the solubility in the SC (Ssc). the overall saturated flux is given by a solubility of different solutes.3 is that it really only reflects where low solute concentrations are used. saturated) solute flux from aqueous vehi- log Ksc and log P (Roberts et al.e.w + log S w = . v ´ Sv and the relationships between rately. Thus. the usual solubility of the solute in the vehicle Hence. suggests the slope for the linear relation- both SC-water partition coefficients and in water ship for a large data set of lipophilic solutes is . 4. flux÷vehicle concentration defined by free volume and other theories (Kuswahyuning and Roberts 2014) and is lim.v between the SC ties by the components of the vehicle.e.6) Thus. it should be an invariant in the = sc K sc . A difficulty in relationship. Further. in turn. The cient between vehicles. we showed that k p = K sc . We have previously applied the main determinant for maximal (or more accu- Ssc = K sc . epithelial permeability in biological systems A limitation of Eq. the depen- cles is the solute size (molecular weight or dence of solubility of the solutes in water on the molecular volume) with an r2 of 0. in accordance with the theoretical importance of solubility is evident variation in solute solubility in the different when it is assumed that the SC is the main barrier vehicles (Roberts 2013). they may determinants for skin penetration: diffusivity in affect dermal blood flow. noting approach defines transport in terms of a perme.5 that the solubility 2009.0.0. as most vehicles ute flux in expressed terms of its underlying do in fact affect skin permeability. 2004.

0 our data (Zhang et al. 4.5 2. in recognising SC solubility as a determi- nant of flux. concluding that propylene glycol data from (Hinz et al.760. 0 1 2 3 4 5 We also showed a parabolic relationship log P between the SC solubility and log P for phenols and that the saturated flux versus SC solubility Fig. showing mal membranes.0 maximum flux and solute lipophilicity in vitro in log [Jmax (%dose/h)] humans (Cross et al. symbols) and non-steroidal anti-inflammatory drugs We then extended this study by using a pro. We and others have shown that there is a parabolic relationship between the saturated solute flux for such solutes 0.5 3. slightly higher than the a 2.2. similar diffusivity). Zhang et al. 1986. exp et al.3) to 0.” Indeed. 150) applied in aqueous solutions to excised human epider- diffusivity for these similar size solutes.69 (Wang et al. 2011). 2.. that this saturated flux – log P relationship was (b) log (% absorption over 4 h) for salicylates (closed defined by a variation in SC solubility with log P. 4.3). (open symbols) applied to human skin in vivo. Original data from (Yano et al. (Original data from Zhang et al. 4. 2009). (2012) are also advocating the inclusion 1.0 and log P (a measure of solute polarity) (Yano 0 1 2 3 4 5 et al.5 slope of 0. 1986.0 3. The curves are for illustra- containing vehicles enhanced the maximum flux tive purposes only . Hinz et al.2  Convex dependence on log P seen in (a) log was linear (Fig.0 2. 2009). 2001). Bucks and Maibach 2005). 2010). 1991) and hairless mouse skin (Hinz et al.5 aqueous solubility for some solutes.est (µg/cm2/h) solubility model used by Yalkowsky and col- leagues (Yalkowsky and Valvani 1980) and com. 1.0 Fig.e.57 we found for our data set. Magnusson et al.0 of the total polar surface area in estimation of 1. Ali et al. hairless rat (Diez et al. 1986) and (c) log Jmax for a series pylene glycol-water co-solvent vehicle (Zhang of phenols across hairless mouse skin in vitro (Original et al.0 mented: “The final conclusion of this study must be that predicting aqueous solubility is indeed still a formidable challenge! The results obtained 1. 1991. log P 2006. Parabolic relationships seen in 0. Several other studies have shown a parabolic relationship between c 2.5 in this study clearly indicate that no one model was able to predict solubility accurately.0 1991) and in vivo (Yano et al.0 point (MP) and hydrogen bond acceptor ability increases the r2 for the prediction of saturated log % Abs (0-4 h) fluxes (Eq. Magnusson log P. The effect of solubility will be most evident 1. showed that inclusion of melting b 2. 4. 1991).0 for a group of solutes that are approximately sim- ilar in size (i. 2009) with epidermal membranes and that of Yano (humans in vivo) and Hinz (hairless mouse skin) are illustrated in -1.4  Non-formulation Parameters Affecting Skin Permeation 51 0. with a log P independent Jmax a series of similar-sized phenols (MW approx. Hewitt et al. (2009) recently evaluated the generalised log Jmax.

 4. (b) log Ssc vs log P.3 + 0.81). r2 = 0. MW and MP (n = 208. vehicle can be expressed as a fractional solubility The second approach is to examine skin perme.5 2. these have been represented by a out that when the fractional solubilities for the combination of organic – water partition coeffi. 2012). examined the in vitro skin fluxes from the 2010.0102 ( MP . Jmax vs log P. i. 4.5 1.0 log Ssc (µg/mg) 1.5 3.4 log P. in a simple or complex vehicle. The most comprehensive Despite much effort in developing quantitative analysis of kp in terms of solute size and lipophilic. not affected the skin or its extent of skin perme- fusion and partition coefficients (Eq.5 2.6 .0.5 log Jmax.0 2. 52 J. 1971).0 2.e.0. r 2 = 0.7) approaches and human skin donors.219 log P .0 2.8) . 2008. it may be applied to other ­showing that diffusivity was not dependent on the vehicles if the concentration of the solute in the lipophilicity of the solute.E.5 1. Raoult’s Law is ation in terms of the skin permeability coefficient approximately obeyed and the vehicle has either (kp). which is defined by two main parameters: dif.0 1.7: risk assessment purposes can be further improved. in which variability is likely to have . but again aqueous solutions.sat ´ f v (Roberts 2013).71 log P mental data.sat They concluded that the donor concentration.81) (4. solute size and solute polar- log J s . The curves are for illustrative purposes only of these solutes by increasing their solubility in Whilst this model probably is currently limited to the intercellular lipids of the SC. (c) linear relationship bility (Ssc) and log P (experimentally determined) for a between log Jmax and log Ssc (Data adapted from (Zhang series of similar-sized phenols applied in aqueous solu.5 3. et al. Grice et al. structure–activity relationship (QSAR) models. J s = J s . (a) log estimated from a 10 % solution). 4.8 for estimation of Js.3  Relationships between maximum flux (Jmax.0 2.2 1.est (µg/cm2/h) log Jmax (µg/cm2/h) 2. from log P. 4. 2012) estimation of kp (Lian et al.67 ) (4.1.5). but also the range of model for saturated skin fluxes by combining the various experimental and vehicle conditions Potts Guy (Eq. EDETOX database in their regression analyses in Milewski and Stinchcomb (Milewski and terms of not only the molecular descriptors of the Stinchcomb 2012) have arrived at a predictive penetrants and the vehicle. vehicle f v ( = Cv / Sv ) and skin f s ( = Cs / S s ) can cients (presented by log P) and size (represented be assumed to be equal.5 1. skin group (Yalkowsky and Valvani 1980) for water ­pre-­hydration. occlusion and the skin thickness. stratum corneum solu.sat = 4.0 1.0061 MW arisen from the use of different experimental ( n = 93.0 2.5 2. tions to excised human epidermal membranes. as shown in Eq 4. In a recent work. exp log Ssc Fig. (Samaras et al. ation enhancement is known. solubility to yield Eq.6 2. A key issue is the large set of historical experi- log k p = .5 1. the skin flux is given by by molecular weight MW or molecular volume Eq.0 1.25 ) ( n = 208. Samaras et al. 2009). Chen et al.5 1.0.  4. We have pointed Traditionally. 2. ity was presented by Potts and Guy (Potts and Guy the predictability and quality of these models for 1992).0 3. r 2 = 0. Moss et al.5 2. MV) (Lien et al.0.0086 MW .0 3. solute lipophilicity.6. exp log P. In an attempt to cover as many formulations and conditions as Many others have since attempted to improve possible.7) with that from the Yalkowsky used – including finite/infinite dosing.

route through the skin (Fig. However. This pathway The in vivo follicular penetration of caffeine has previously been considered as insignificant was also systemically examined using pharmaco- as appendageal structures only make up 0. hair follicles and their associated piloseba. The and/or reservoirs have been increasingly explored in vivo absorption of caffeine was fast and high and received more attention. Mitragotri et al. molecules (Essa et al. 2002. (2007) cellular pathway) and intercellular lipids (inter. the “porous pathway” model was introduced to The infundibulum increases the possible surface address this challenge by describing skin per. The “brick-and-mortar” model has Work by a number of groups has found that also been adapted and expressed in different the follicular route predominates for hydrophilic forms (Wang et al. providing target sites for pen- distribution. tional absorption.5 Physicochemical Properties minimal entry into the follicular pores. 2011).1–1 % kinetic modelling (Liu et al. A para- included. the bulb region. 2004). In this antigen-presenting cells. whereas the lower bulge model. though its pathway” or “shunt pathway” by appendages molecular size is slightly smaller than the above such as hair follicles also acts as a permeation penetrants (Essa et al. There was a (forearm to forehead) of the total skin surface 10 times faster absorption from hair follicles. A study by Frum et al. gland. had 4. and this was nanoparticle delivery (Lademann et al. Lai and Roberts 1999. Tang et al. estradiol and corticosterone. Wang provides decreased barrier properties and is sur- et al. 2002. Mitragotri 2003). The lower infundibulum lipophilicity alone (Nitsche et al. considered. For man- nitol. Consequently.05). as well as the vehicle melting and boiling The pilosebaceous unit comprises four differ- points were prominent factors influencing skin ent areas for topical application: the sebaceous flux. mal pathway easily. whereas a That Favour Appendageal 34 % contribution to total skin flux was found for Permeation hydrophilic adenosine (log P = −1. bolic profile shows a maximal follicular contribu- tion occurs at log P around 1. hydrophilic solutes are assumed to dif. the physiological differ- . the “bulk ated with its flux through human skin. quantified the influence of the drug lipophilicity cellular pathway). 2006. skin porosity and tortuosity are etrated molecules (Knorr et al. resulting in a microenviron- philic and polar solutes penetrate through the ment that is rich is neutral.e. 2002. 2002). 2007. 2008. This mathematic model aims to include Mitragotri 2003) that cannot permeate the epider- permeation through the corneocytes (i. area for targeted delivery and is an area of addi- meability to hydrophilic solutes (Peck et al. 2010.4  Non-formulation Parameters Affecting Skin Permeation 53 ity. 2005). corneum. chain fatty acids. The two most lipophilic drugs. 2011). there was a 54 % follicular contribution associ- and intracellular permeation routes. the SC is a heterogeneous mem. which is also hydrophilic (log P = −2. Chen et al. 2007. and area (Otberg et al. The immediate region sur- 1994. 4. Some hydro. trans. 2009).47) As an alternative to the stratum corneum inter. rounding the infundibulum is rich in Tezel et al. 2006. by in vivo blood flow. with the follicular route contributing ceous structures acting as a permeation pathway 25–35 % to the overall skin absorption. 2013). considered to be mainly due to a fast resorption Patzelt and Lademann 2013). in recent a 10-min delay for transport through the stratum years. 2001. especially for compared to the in vitro absorption. the hair matrix cells and In reality. non-polar lipids SC faster than would be anticipated based on (Meidan et al. the hair follicle infundibulum. rounded by a dense network of blood capillaries. where the geometrical and on follicular penetration by using a range of compositional parameters of the SC have been drugs with similar molecular weights. where pore size Lademann 2013). Sebum excreted brane consisting of several layers of corneocytes by the sebaceous gland is composed of short-­ with the space filled with lipids. Ogiso et al.1). region is surrounded by stem cells (Patzelt and fuse through pores in the skin.

solubility parame- remain in the superficial regions of the follicle ters and charge) (Trommer and Neubert 2006). Grice et al. the transfollicular (Lademann et al. 2006). . Vogt size-dependent manner (Patzelt et al. 2011) (see Fig. which may be solid or vesicular. sion from Patzelt et al. whereas. Topically applied poly. pathway is also an important route for nanopar.e. found that antigen-presenting cells sur- 643-nm particles penetrating deepest into the fol. 4. 4. where both particles and dye molecules ties such as size. may be ideal follicular delivery (Lademann et al. thus pro- Lademann has suggested that the optimal size viding a promising mechanism for vaccination of particles for deep penetration is governed by via the follicular route. 2007). In addition. ticles. 4. which may be targeted to deep follicular with therapeutic substances bound to the surface regions and function as drug delivery vehicles or encapsulated. 2008). overlapping cells (cuticula pili) (Reproduced with permis- minal hair follicles (THF) and vellus hair follicles (VHF). to a maximum depth of approximately 40-nm particles that have permeated through the 1100 μm (see Fig. which may by manipulating the formulation (i. follicular epithelium (Vogt et al. agents. 2011 and Lademann et al. miscible ingredients) or modifying the target 2009). The effect of massage is demonstrated in molecule (optimising physicochemical proper- Fig.E.4). massage drives the a contraction of hair follicles of the excised skin. particles – but not the dye – deep into the follicle Besides polar molecules. showing the layers of of PLGA particles in relation to the target sites within ter.5. Particles. because of their long residence time meric particles were shown to penetrate porcine within the follicle and their targetable penetration skin with massage via the follicular pathway in a depth (Lademann et al. the size of the cuticula on the hair shafts Targeted follicular delivery can be achieved (Lademann et al. particle-/ act as “geared pumps” to drive the particles into vesicle-based dosage forms and using sebum the follicles during massage (Lademann et al.54 J.4). 2007). 2011) ence in hair follicles of the investigated skin and without massage.4  Above: Penetration depths in μm of different sizes Below: The hair surface structure. with et al. 4. lipophilicity. 2011). Fig. rounding the infundibulum are able to internalise licle.

5  Superposition of transmission and fluorescent and (b) dye in non-particle form – with massage.4  Non-formulation Parameters Affecting Skin Permeation 55 a b c d Fig. 4. demonstrating the in vitro penetration of the dye-­ Particle form and (d) non-particle form – without massage containing formulation into the hair follicles of porcine (Reproduced with permission from Lademann et al. (a) Dye in particle form . (c) images. 2007) skin after application of massage.

6 Physicochemical Properties drug’s lag time into deep dermis. flucon. 1967. Subcutaneous and Deeper Tissue Permeation 4. 2013). NMF functions as a plasticiser of the tion is the size of the solute (Singh and Roberts stratum corneum under basal conditions (Laden 1996. Various human biopsy (Rabinowitz et al. 4. Jokura et al. Brunner controller of hydration is natural moisturising et al. Some other data and concluded that highly plasma protein methods like fractional laser and massage could bound drugs. In addition. with the bricks show that the main determinant for the penetration being likened to NMF-containing keratin of solutes into deeper tissues after topical applica. Transdermal delivery can also be controlled by Grundmann-Kollmann et al.6) and that these particular properties for an optimum follicular transport routes may dominate in some cases. An example reported by Grice et al. lidocaine. 2003). in general. Lee et al. this was likely due to nential concentration gradient. and decrease the 4. The cutaneous vascular appears to be impli. cated in the penetration of solutes in studies con. of water or under conditions of high humidity. subcutaneous That Favour Direct and muscle tissue. networks of spherical particulates are found in salicylic compounds and trolamine salicylate. the concentration that formulations that contained ethanol could gradients for highly protein bound drugs for enhance the early minoxidil uptake into the hair deeper tissue penetration tended to be shallow. 2002. Absorption cal application to specific areas of the body. 2007. may be transported in deeper tissues follicles (Lademann et al. salicylic acid. sponges. 1998. Anissimov the extent of skin hydration. in addition to normal diffusive also selectively enhance permeant targeting to transport. Cross and Roberts 1999. A major (Cross et al. 1995). 2005) studies have been used to show direct factor (NMF). follicles.7 Skin Hydration There are now a large number of studies showing and Percutaneous that deep tissue penetration can occur after topi. The solutes studied were of similar neocytes (Roberts et al. regions where the NMF concentration is at its Dancik et al. ethanol. The original bricks and mortar and deep tissue penetration of solutes after topi. 1992. ibuprofen. Swelling is at its greatest in Monteiro-Riviere et al. They applied a the partitioning of ethanol (plus minoxidil) physiological pharmacokinetic analysis to this through the lipid rich compartments. the corneocytes. Such vascular transport can increase deep tissue concentrations beyond those resulting from extravascular diffusion alone. Cross et al. methyl salicylate. 1982.56 J. swol- size and included diclofenac. Benfeldt et al. nicotine. len corneocytes and separation of lipid bilayers azole. water is able microdialysis literature on the penetration of predominantly located either in the intercellular topically compounds into subcutaneous tissue regions in separate domains or trapped in cor- and muscle. tially and it is now described as being a continu- Rat wound deposition studies have been used to ous poly-­proteinaceous structure. 1999. the NMF-bound water assists in the swelling of ducted in rats. 1993. Kretsos et al. pigs and man (McNeill et al. (2012) combined experimental highest (Bouwstra et al. 1999).E. 2008). that when the skin is highly hydrated. As minoxidil is six times more soluble whereas the poorly bound drugs showed an expo- in ethanol as it is in water. in the intercellular space to form cisternae and 5-fluorouracil. Grice et al. Following the addition 2008). via the blood and lymphatic circulation as well as The importance and required molecular and via interstitial convection (Fig. propranolol. description of the SC has been refined substan- cal application. It has been shown percutaneous permeation studies with the avail. delivery are yet to be fully investigated. (2010) shows They observed that. which mainly refers and Roberts 2011) or cutaneous microdialysis to the extent of hydration in the SC. porcine skin after a 4–10-h hydration period .

(a) Blood vessels.6 Physiological pharmacokinetic model for topical drug transport processes in deeper skin.7 shows high magnifi. keratinisation rate and the type of moisturis. 2010). 2008). mainly influenced by environmental humidity. just as disturbed skin barrier function has cation cryo-­ scanning electron microscopy been correlated with low SC hydration (Proksch images of human SC in various hydration states. lymphatic vessels and interstitial convection in dermis. effects on the transdermal delivery of penetrants The hydration state of the stratum corneum is by altering skin physiology. 2003). increasing water pools present at high levels of hydration the water content of the skin can have substantial (Bouwstra et al. transepidermal water loss (TEWL) the SC can be greatly increased. In clinical practice. showing clearly the swollen corneocytes and In the presence of an intact barrier. by which the hydration of temperature. The inverse relation. 2012)) (Tan et al. drugs (Hafeez and Maibach 2013). occlusion of the skin. et al. the concentration–time profile in the superficial microdialysis probe being used as an input to predict the deeper microdialysis probe concentration–time profile (Reproduced with permission from (Dancik et al. 4. Figure 4. is widely used to rate. (b) Schematic of physiological pharmacokinetic model used to analyse human cutaneous microdialysis data at two depths. 2010). A number of ship between TEWL and SC hydration is well factors could contribute to the increased permea- .4  Non-formulation Parameters Affecting Skin Permeation 57 Fig. enhance the penetration of topically applied ers (Vergnanini et al. known.

upper and lower part. Firstly. The spaces between the lope. This network is surrounded by the cornified enve- mately 360 nm in thickness. 2003) bility. Arrows indicate pools are present a close cell to cell contact is observed. possibly caused by drying water pools are observed (see white arrows). it is likely to be due to an increase water in the stratum corneum. Between the cells frequently large and very small corneocytes are mostly air-filled. (b) SC compared to dry SC or SC hydrated to approximately 20 % hydrated to 17 % wt/wt reveals a low contrast image. Arrowheads indi. (d) A high magnification Arrowheads indicate the cell boundaries.7) could disor- increases the partitioning of the solute from the der the structures of lipids and proteins vehicle into the membrane. In the microscopy images of SC hydrated to various levels. If no water of the SC required for low hydration levels. also illustrated by in the solubility of the permeant in the SC.E. Recent work using Raman spectroscopy meation by several orders of magnitude as previ- demonstrated that increased amounts of unbound ously described (Roberts 1991. permeation rate of topically applied solutes. Roberts and . 4. thereby altering the that the increased hydration could lead to swell. 4. the appearance of the SC is similar to Arrows indicate undulations or interdigitations of cells. T.58 J. This indicates the absence various cells is noticed. The cells are approxi. Grice et al. simi. Changes in skin hydration may affect skin per- ids. (c) SC hydrated to 70 % wt/wt reveals in the indicated by short arrows (Reproduced with permission central part slightly swollen cells with a higher contrast (see from Bouwstra et al. ing of the structure and rearrangement of the lip. of a fully hydrated SC sheet. 2013).7 High magnification cryo-scanning electron black asterisks) indicating the presence of water.electron microscopy (Fig. The undulations or interdigitations of cells. Another possibility is (Vyumvuhore et al. depicted. The cells that are less swollen are of water pools. that of dry skin (white asterisks). The keratin network is clearly acterised by low contrast images. tissue-­freezing medium. This cryo. (a) Dry SC char. cell ends are round and fewer undulations are observed cate the cell boundaries. a b c d Fig. A difference in the degree of swelling between the lar to that observed for dry skin. wt/wt.

b Taylor et al. the more lipophilic steroids tion of solutes with varying octanol water partition coeffi- appear to have a greater enhancement by occlusion cient (log P) across human skin in vivo: (a) Phenols. open symbols occluded. methylethylketone. 50 % Dose absorbed rin.8). (b) than the more polar ones (Fig. facial concluded that occlusion was more effective in cleansers. 30 Interestingly. Occlusion is not the only means of increasing fen delivered from ultra-deformable. lipophilicity and occlusion-induced permeation For example. has good water-binding and exfoliating proper- pounds than in relatively hydrophilic compounds. Cevc et al.8  Occlusivity does not uniformly enhance penetra- ined. 4. ketoprofen was promoted by occlusion. the permeability of topically applied free the intercellular regions (Fatouros et al. In the same study. Evidence shows that increasing a 70 room humidity can lead to a marked increase in 60 absorption for a range of solutes including aspi. Chemical and physical enhance- transfersome carriers by apparently eliminating ment methods can also affect skin hydration. (Roberts and Walker 1993). iontophoresis may hydrate the stratum drives the transfersomes across the skin. but 40 with little influence on the percutaneous absorption 30 of glycerol. where a series of phenolic log P compounds with varying lipophilicity was exam- Fig. corneum due to the formation of water pools in trast. current findings suggest that there are no quantitatively defined relationships 20 between hydration-enhanced skin permeability 10 and the lipophilicity and molecular structure of 0 the solutes. 4. steroids. prevent water loss and have a softening and cally valid QSAR developed from an extended soothing effect on the skin. (2002) found that occlusion of the 70 skin affected the skin penetration of linoleic acid 60 and glycerol differently. occlusion suppressed the permeation of ketopro. A similar result was found by Bucks 1 2 3 4 and Maibach (2005). various cor. 2006). (2008) found that skin enhancement could be discerned. For example. For the transcutaneous water gradient that normally instance. closed symbols unoc- It should be noted that the effects of occlusion cluded (Data modified from (Bucks and Maibach 2005)) on skin permeation are not clearly defined or uni- form (Roberts et al. Work by Hikima and Maibach (2006) 20 showed a random distribution in human skin fluxes of steroids in relation to their lipophilicity or 10 molecular weight when the skin was either hydrated 0 or dehydrated. 2007). Samaras Water-binding agents (humectants and or hydrat- et al. although the early study by Wurster -10 and Kramer (1961) suggested that hydration 0 1 2 3 4 preferentially enhanced the skin penetration of log P the more polar salicylate esters. Some but emphasised that no clear relationship between studies have also shown that urea may act as a . bath oils. (2012) further showed that skin occlusion ing agents) are also classified as moisturisers that had a significant non-linear effect in their statisti. enhancing permeation of very lipophilic com. In con. with occlusion suppress- % Dose absorbed 50 ing the delivery of linoleic acid from an ethanolic vehicle compared to the unoccluded condition. Hafeez and Maibach (2013) recently ingredient in some hair conditioners.4  Non-formulation Parameters Affecting Skin Permeation 59 Walker 1993). skin softeners and lotions. 2004. hydrophilic skin hydration. fusidic acid. Pellanda et al. 40 ticosteroids. an database. etc. urea. ties for skin when used in small amounts.

The skin pH et al. however. as were the pH Beastall et al. pH. Pre-term babies environment for the diffusion of topically applied have elevated levels of both TEWL and transcu. No TEWL. Capacitance values. as it appears that skin used as a disinfectant on the skin of pre-term pH is constant throughout adulthood to approxi- infants. but period of 30 weeks or less. Similar variations Giusti et al. measured the TEWL. assumed that aged skin provides a more variable is complete in utero by 34 weeks. the effect of aging on blood flow.60 J. penetration enhancer (Wohlrab 1984. thickness Ultra-low birth weight infants from the ages of and ultrasound echogenicity (Waller and Maibach 23–25 weeks were shown to need at least 4 weeks 2005). thickness of the skin layers seemed to have been late through ex vivo newborn infant skin. elastin 4. taneous heat transfer. certain structural changes etrate into the stratum corneum. 1998. acellular and avascular. Humectants with a small molecular age. as discussed below (Byl 1995). compared to full term remains constant in areas such as the buttock. 1986). Grice et al. 2001). and pH of the volar forearm and buttocks of 70 TEWL has been used as an indication of the infants aged 8–24 months and compared these to effect of aging on barrier function. showed that when hexachlorophene was data were more convincing. They point out that an alter- quantify the amount of water in the SC. the pH increased. infants (Barker et al. urea and glycerin) are assumed to pen. As the skin ages. dermal absorption occurred which mately 70 years of age. found to be higher in infants. The change in et al. After 70.E. to interpret. whereas babies described age-related changes in skin structure of 30–32 weeks had barrier functions comparable and function were often conflicting and difficult to adults (Kalia et al. very readings. also have humectant tions were observed in infants according to sex or properties. Exactly Performance how the penetration barrier of the skin changes with age remains unclear. 1975). so that the aged SC is considerably drier role of natural moisturising factors to retain than that of a young adult. Fluhr et al. con. 2007).8  ge-Related Skin Barrier A and glycosaminoglycan are also altered. are Waller and Maibach reviewed the literature on largely dependent on how premature the baby is. it should be Epidermal development. 1987). 2004). dorsal forearm and shoulder. which as expected. resulted in myelinopathy in the premature infants which was suggested to be due to stasis of blood. The epidermis becomes thinner and 2008). (Anderson et al. Collagen. found that absorption of the compound was 100– The epidermis thickens with age on certain sites 1000 times higher in infants who had a gestation such as the volar and dorsal upper arm. including SC structure. Another study by Barker especially in the lower limbs. capacitance were noted for dermal thickness. especially in photo-aged areas. the keratinocytes become less adjacent to one another.g. The cutaneous blood flow results It has been widely reported that premature suggested that there may be a trend towards infant skin is more susceptible to penetration than decreased cutaneous perfusion in older individu- fully formed infant skin. They concluded that the data which to develop a fully functional SC. As and Wickett noted considerable variability in expected. it has a reduced lipid moisture (Caussin et al. It was influenced more by site on the body than by age. A study by Anderson als. which and Wickett 2008). the content and frequently has a less extensive micro- mode of action is still poorly understood (Loden circulation. Glycerin based creams. However. capacitance or pH varia- familiar to most consumers. size (e. The dermis also becomes relatively atro- phic. no differences in TEWL were found TEWL with age in Chinese female women living between the infants and adults at either site. drugs. 1990. examined the permeation of sodium salicy. with a quadratic regression showing firming that the infant skin had a fully developed little change between 10 and 70 years (Hillebrand barrier function. in Beijing. mimicking the occur. Hillebrand values in adult skin (Giusti et al. were native explanation for these findings is that the .

below). BA. (2013). As aged skin is drier. it seems that two main age-­ administered topically to the ventral forearms of related changes in barrier properties of skin are volunteers. benzoic acid (BA). Similar findings of a minimal effect of absence of changes in TEWL (Rougier et al. acetylsalicylic acid. The effect of age is patients. compared to modification) could manifest slower release younger subjects (20–30 and 45–55 years). acetyl. the SC surface becomes function with regard to TEWL. The time it took for water to jects. reduced water content also favours the perme- hydrocortisone (HC). ation of more lipophilic molecules. with compounds are. versus log P of the drug. The permeation of HC. changes in percutaneous permeation have been The authors concluded that reduced lipid content reported. In neocyte surface area (see Sect. it requires more water to while the absorption of both of the most lipophilic achieve a fully hydrated state. expressed as the ratio of the younger group (age 19–42) than the older responses in old (>65 years) and young (22–40 year) sub- group (age 69–85). Roskos and Guy concluded that the ability 20 of the SC to control evaporation from the skin is not age-dependent and its ability to prevent dehy. less water may have been required (Data modified from (Roskos et al. exercise) depending 80 on where they were born. diet. 0 dration is also maintained as a subject ages 0 0. affected by the age of the volunteer’s skin. estradiol (EST). showed a similar age-related intercellular lipid organisation. to test whether permeation through Accordingly. These the same study.5 1 1. Testosterone (TST). the et al. term babies and moderate change in skin barrier .5 (Roskos and Guy 1989). young different stresses (sun. work agrees with the previous findings showing Despite the minimal effects of age on barrier that. are caffeine. in the kinetics. The subsequent decrease in benzoic acid permeation in older dehydration (and reorganisation of this structural male subjects (65–80 years). Drugs. 9. which appeared related to changes in cor- were reported by Luebberding et al. Roskos et al. the more their permeation is the lowest skin surface lipid levels seen in sub. As older skin has been shown to have reduced water content. the Fig. testosterone. In 60 another study of adults aged 19–85 years show- ing no correlation of ventral forearm TEWL with 40 age. the percutaneous permeation of the aging skin varies across a range of penetrants with more hydrophilic molecules is more affected than different physicochemical properties (Roskos lipophilic molecules. benzoic acid. in order of increas- diffuse through the SC was longer for older ing log P. examined the penetration of in aging skin results in a poorer dissolution a range of drugs in young (18–40) and old (>65) medium for topically applied chemicals. age on TEWL at six different body sites in women 1988). When the SC barrier log P was deliberately perturbed by occlusion. hydration-­induced structural alteration of the Rougier et al. salicylic acid (ASA) and caffeine (CS) were To conclude. 4. ASA and expected: decreased barrier performance in pre-­ CS was significantly lower in older volunteers. Cumulative % water activity gradient was lost more rapidly in dose absorbed for six drugs. which may dominate any relationship between TEWL and skin age. significant decreases in sebum results indicate that the more hydrophilic the production were seen with increasing age. 1989)) for the SC to achieve the fully hydrated equilib- rium state. volunteers. It is possible that compounds TST and EST was similar in the older occlusion in the older subjects produces a and younger volunteers (see Fig.9  Percutaneous permeation data. This jects over 70 years old.9). estradiol.5 3 3. age-related drier and sebaceous gland activity is reduced. 4. as the skin ages. hydrocortisone.4  Non-formulation Parameters Affecting Skin Permeation 61 women in the different age groups experienced 100 % response: old vs. 1989). It was explained that when the tissue most pronounced for the more hydrophilic compounds was occluded.5 2 2.

1987) have ties of the permeant. depend on the physicochemical proper. and other chemicals (Rougier et al. in which materials applied to plantar and palmar skin. gested that these cellular variations could A key determinant for differences between sites explain previous observations of regional dif- is SC thickness. Early work by Plewig and Marples showed significant Site-related or regional variations in SC structure regional differences in corneocyte dimensions. As noted previously. and shown (Scheuplein and Blank 1971). there are also a been reported. pesticides (Maibach et al. (1988) excretion following topical application of methyl examined the effect of corneocyte size on ben- salicylate on five separate body sites for 10 h zoic acid permeation as well as TEWL in (Fig. Results from the Roberts group est number of cell layers and the greatest showing 48-h cumulative urinary salicylate TEWL. smaller corneocytes can pack more closely compared to other sites. especially for eters skin (Marrakchi and Maibach 2007. humans (Rougier et al. 1971) however.9  ite-Related Skin Barrier S made to relate the regional variations in skin Performance permeability to skin properties. it has been an excellent correlation between the reciprocal suggested that plantar and palmar SC should be of path length and TEWL (Machado et al. Recent work from Hadgraft’s laboratory (Rushmer et al. performance in elderly. showed a good over any underlying apparent relationship correlation between benzoic acid permeation between skin permeation and aging.E. Scheuplein and Blank 1971). 2012. explained by corneocyte size alone and that tions in TEWL. Grice et al. This work also meation behaviour of certain types of molecules showed the forehead to be the site of greatest per- through aging skin. If 1/path . 1988). lag times. In addition to mapping. Maibach 1967).10) clearly show the effect of increased humans and concluded that regional differ- SC thickness (Roberts et al. 2013) and the per. Rougier et al. are well recognised and may be reflected in dif. However. Kleesz corneocytes <600 μm2 or >1000μm2. the small- Blank 1971). ysis of Rougier et al. the effects of which. despite its having greater permeability have calculated the intercellular path length to water than abdominal skin. Machado et al. As seen in data for water together. Luebberding et al. arms. resulting in a shorter intercellular permeation quoted by Scheuplein and Blank. 1982). The differences in thick. Machado et al. and abdomen 1967). A re-anal- et al. hydration and other skin param. Earlier work by Rougier et al. Scheuplein and the smallest corneocyte surface area. attempts have been 4. other factors were important. legs. considered separately from the thinner horny lay. the forehead had ers at other sites (Kligman 1964. ness are manifested in delayed responses to 2010a) has suggested a mechanism. the path length for permeating molecules to tra- greater thickness of plantar skin leads to longer verse the layers of corneocytes. (Hadgraft and Lane 2009. 2010a). with scrotal. The increased lag from the corneocyte surface area and the num- time effectively masks the increased permeability ber of cell layers at six body sites. Indeed. A combina. which can vary from 400 to ferences in TEWL (Baker and Kligman 1967) 600  μm for the plantar and palmar callus to and hydrocortisone (Feldmann and Maibach 10–20 μm for the back. forehead and wrist number of confounding environmental and skin generally more permeable than skin on the genetic factors at play and these may dominate trunk and periphery. meability for water loss (scrotal skin was not examined). 1966).’s data for benzoic acid meability of hydrocortisone (Feldmann and permeation supports this conclusion.62 J. The authors sug- and Maibach 1967. with the smallest cells found in the forehead ferences in permeability of the skin (Feldmann (Plewig and Marples 1970). may be responsible for the per. 4. ences in permeation could not be entirely Various studies mapping the regional varia. (measured as 14C-benzoic acid excretion and tion of factors. including reduced lipid and water TEWL) across seven different body sites in content of the SC.

98) is very strong. forearm and calf. 1984). their similar- tion in seven body sites (Otberg et al. 1999) and the volume and surface area of the follicular infun. 2013) impacts on the perme- and permeation results for four body sites. since it of each solute was found to correlate inversely had been accepted that hair follicles made up with the percentage lipid content in each piece of less that 0. with topical application of the vasodilator ben- port and that variations in follicular size and zyl nicotinate. although decreasing or artificially while that for benzoic acid permeation is increasing the level of sebum on the skin surface weaker (R2 = 0. flow. 1971. cal properties. found regional differences in et al. variations in cutaneous blood distribution may contribute to regional varia. 2006). (Otberg et al. 1987. Elias et al. This may be considered unlikely. 1987. 2004) Elias and Brown may have est in the forehead and least in the forearm. forehead skin surface (Rougier et al. 4. 1988) and chemical permeation Laser Doppler techniques allow the mea- (Feldmann and Maibach 1967. likelihood of a similar contribution by follicles dibula per square centimetre of skin was great.75). layers (Elias and Brown 1978). 1984). Rougier et al. to the forehead (Wester et al. These correlations indicate basal skin blood flow across the forearm that the follicular route may contribute signifi. (Johnson et al. surement of blood flow to surface tissues. Maibach Based on their results from permeation of explained the relatively high hydrocortisone water and salicylic acid across abdominal and leg permeation at this site by suggesting that a sub. have suggested that the regional stantial amount of permeation was probably variations in skin permeability arise mainly from occurring through the follicles rather than the differences in the lipid content in the epidermal SC (Feldmann and Maibach 1967. 1982)) 150 100 50 0 plantar heel instep forearm abdomen length is calculated and plotted against TEWL Luebberding et al. while in kinetic studies cantly to chemical permeation and water trans. However.4  Non-formulation Parameters Affecting Skin Permeation 63 Fig. centration of sebaceous glands. It is not clear whether the forehead. Scheuplein and Blank 1971. 1988). has no effect on transepidermal water loss Noting that the forehead contains a high con. The forehead had a higher blood flow and . the or the SC thickness. Maibach et al.1 % of the surface area of the skin skin but did not relate to the number of cell layers (Schaefer and Redelmeier 1996). The ity in SC thickness (Ya-Xian et al. with been able to reveal a dependence on SC lipids in the overall pattern of follicular distribution the absence of other strong contributing being similar to that seen in variations in TEWL differences. (Rougier et al. both basally the significantly greater sebum content on the and in response to the drug (Jacobi et al. 2004). Rougier Johnson et al. skin. The permeation 1987). the ation of molecules with different physicochemi- correlation for TEWL (R2 = 0. Given the relatively low per- Lademann group has identified significant meation of chemicals at these two sites compared regional variations in follicle size and distribu.10  Cumulative urinary salicylate excretion Cumulative salicylate excretion (mg) 250 showing the effect of stratum corneum thickness at five different body sites on the percutaneous 200 absorption rate of methyl salicylate (commercial 25 % formulation) applied to a 50 cm2 area for 10 h (Data modified from (Roberts et al. temperature and redness were seen across tions in skin permeability. (Kligman 1963).

groups (Fluhr et al. (a) TEWL cutaneous blood flow determined by Laser Doppler flow- (Rougier et al. 2004). post-menopausal women on no hormone replace- mulations or any material which comes into ment and 25 male subjects found that the skin of contact with the skin is substantial and depends pre-menopausal women contained significantly on different variables as discussed above.E.10 G  ender-Related Skin Barrier knowledge on regional variations in skin perme.64 J. studies In conclusion. expressed as metry (Jacobi et al. (d) ties of human skin based on rank order. This smaller corneocytes compared to the other two implies that for optimised delivery. and the number of cell layers (Machado et al. the influence of the site of appli. (e) skin permeation of hydro- infundibular surface area mm2 per cm2 of skin (Otberg cortisone (Feldman and Maibach 1967) and parathion et al. As far as the skin structure is concerned. normally elevated in pre-/ post-menopausal a TEWL b Follicle c Path d Cutaneous e Skin Density Length Blood flow Permeation Low High Fig. regional differences in skin permeability in vivo. dermatologi. Performance ation and some of the factors that may contribute to this. 4. Grice et al. These results suggest that differences in on the product if the site dependency in absorp- cutaneous blood flow may also account for tion affects the treatment or safety. based on corneocyte surface area excretion of the 14C-compound over five days .11 attempts to summarise the state of 4. (c) minimum intercellular path length through (Maibach et al. 2005). Figure 4.11  Regional differences in responses and proper. 2010a). the site of application should be specified lator. controls the rate corneocyte area. in 33 pre-menopausal women. The authors hypothe- cal formulations should be designed in a way that sised that sex hormones may have an effect on the formulation. by Fluhr et al. 2004). 2006). and not the skin. greater and more rapid responses to the vasodi. 1971). both measured as total urinary the stratum corneum. As well. (b) follicle density. 21 cation on percutaneous permeation of topical for. since sex hormone levels are of delivery if it is designed for different skin areas.

Following a 24-h application of the function. Accordingly. the above-mentioned studies into the ­circulatory system and its relative toxic- might reveal that among different biological fac.1 Insect Repellents between women and normal-sized men. 1982). However. such as in nifedipine penetration (Kondo et al. 2010a). supports Insect repellent creams. It of age and gender. The absorption of DEET To conclude. result. No change in barrier performance is.) based on gender was examined (Prather found in cosmetic products on skin barrier et al. (1988) reported no increase of the market.4  Non-formulation Parameters Affecting Skin Permeation 65 women compared to the other two groups. it has been 4. A recent review by Machado et al. m-toluamide (DEET).N-diethyl-­ der (Machado et al. N. and until proven otherwise. . insect repellents and sun. lotions and sprays are the above observations. a lipophilic vehicle with enhancer prop- erties used in medications and cosmetics such as Application of products on the skin for cosmetic shaving lotions) resulted in a remarkable increase purposes is widespread. it is equally interesting to note that DEET is also taneous absorption of drugs. Some products. we Nicoderm® transdermal delivery system (Alza will examine the influence of certain actives Corp. when male and female subjects cation of therapeutic agents is sometimes accom- aged 20–60 years were compared (Machado panied by the application of cosmetics. the levels in nico- tine plasma concentration (Cmax) and area under the curve (AUC) did not vary significantly 4. and anti-aging prod. repellents with more than 200 million people ery and that any conflicting results may be due to using products containing various concentra- changes in the skin’s barrier function related to tions of this active (10 % or higher) worldwide the menstrual cycle (Reed et al. all to improve the dermal and transdermal delivery laboratories use mixtures of male and female of a variety of drugs through hairless mouse skin samples for their investigations indiscrimi. 1995). tors. the public for more than 50 years. 2006). (2002) observed a slight increase in indomethacin flux.11 Barrier Performance reported that mixtures of DEET and DES (diethyl in Cosmetically Treated Skin sebacate. would be interesting to discover if the application deodorants. transdermal nicotine system. carried out a study in which the occur in cosmetically treated skin can increase differences in permeation of nicotine from a the risk of dermatotoxicity. 2010b). impaired skin barrier properties that may Prather et al. 1993). finding might indicate that DEET is not an effec- screen creams are used by everyone irrespective tive permeation enhancer when employed alone. This moisturising creams. (Debboun et al. nately and there is no gender-specified product in Kondo et al. and suggests that skin widely used products. a hydrophobic com- cluded that there were no gender-related pound. Cosmetic after the application of insect repellent would products contain ingredients that can alter both increase the penetration of the actives contained in the structure and function of the skin and a the aforementioned cream. differences in corneocyte size were observed by anticipated. is the most common ingredient of insect differences in barrier integrity and barrier recov. ity are topics that have been of interest. having been available to barrier function does not seem to vary with gen. skin (Windheuser et al. 1988). of a cosmetic cream containing DES before or ucts are more age and gender specific. However. therefore. as a considered to be a permeation enhancer. while depilatory creams. Reed et al.11. This is important because the appli- Machado et al. gender seems to have lowest effect on percu. absorption of nifedipine in the presence of DEET while Ogiso et al. antiperspirants. also con. As a et al. In this section. shown rule of thumb.

It has 1943).11. phide bridges contained in corneocytes. treat. Some these intercellular bonds thus compromising the other hair removal methods also can affect skin structural integrity of hair strands.E. tions of phenol or α-hydroxy acids (AHAs). observed in vivo after glycolic acid treatment ment with a depilatory agent produced some highly returned to pre-­treatments values after one day noticeable morphological changes. depila. the systems. 2004).3 Peeling Agents hair follicle and greatly enhances the low-­ resistance character of this pathway (Han et al. 2008). The anti-ageing industry. Song et al. reported that the increase in TEWL ised by tight packing of clearly defined cells. However. are strengthened by intercellular disulphide one would not be able to conclude if continued use bridges formed between two cysteine residues. Corneocytes also contain keratin fila. barrier function of the SC is compromised and tions. In this direction. of depilatory creams induces any long-term struc- Calcium and potassium thioglycolates reduce tural or permeability changes in the skin. also reported changes in SC struc. the permeation of compounds. on the market for hair removal. to the necrosis of the SC and its separation from gests that the structural changes induced by the the viable epidermis whereas keratocoagulation depilatory agent may be restored later. Therefore. turation and coagulation of keratin (Rothman meation enhancement is not well understood. Consequently. Firstly. located within the cornified cells and in the One example is chemical peeling. struc. resulting Shiozuka et al. no studies have been conducted are calcium thioglycolate and potassium thiogly. has an overwhelming number of products ments. As a result. which cell envelope (Madison 2003). 2008). et al. the skin barrier function is been reported that the structural changes induced significantly altered after this procedure. Secondly. Grice et al. to the newly forming layers of the SC. 4. lars. tein. involves treating the skin with low concentra- tory agents will also be able to reduce the disul. Keratin filaments contained in hair strands of calcium or potassium thioglycolate. in the formation of more supple superficial lay- ture after treatment by a depilatory agent (Shiozuka ers (Van Scott and Yu 1984).2 Chemical Depilatories tural changes in the SC were observed and the defi- nition of cell borders was disrupted. Therefore. worth billions of dol- 2004). such as glycolic or salicylic acid to remove the weakening their structure. resistance to transdermal delivery is greatly The two main components of these formulations reduced. involves the irreversible phenomenon of dena- The mechanism of thioglycolate-induced per. 2010). This sug. These acids are known to and lipophilic compounds through rat skin pre-­ cause irritation to the skin by producing one of treated with a depilatory agent was significantly the following reactions: keratolysis or keratoco- enhanced and that this enhancement persisted for agulation (Brown et al. found that superficial layer of the skin. shaved skin enhances the permeation of aluminium bing (Goddard and Michaelis 1934). presenting a A plethora of products and services is available ‘homogenised’ pattern (Lee et al.11. 2008).66 J. the SC and to stim- the transdermal penetration of both hydrophilic ulate its renewal. ranging from the volume of intercellular spaces and gaps high-tech laser removal to the more conventional increased (Shiozuka et al. and treatments promising to turn back the clock. (Song et al. such as shaving and depilatory formula. 1997). 2010). 1961). Keratolysis leads 24 h after application (Lee et al. to date exploring the effects of chronic application colate. Hair removal induces the 4. Lee et al. It has been shown that hair can be easily removed by scraping or rub. that are the subject of the present review. . The absence of the hair shaft is influential in promoting cutaneous absorption through the transfollicular route. salts (Anane et al. which constitute a major structural pro. Whereas the normal SC is character. AHAs by thioglycolate-based depilatory creams enhanced reduce the skin barrier integrity by reducing drug delivery through both the transcellular and corneocyte cohesion and their influence extends intercellular SC pathways (Lee et al. Consequently.

therefore. In addition to the underlying. Hillebrand and 1996). contribute to using excised mouse skin. sodium lauryl sul.4  Non-formulation Parameters Affecting Skin Permeation 67 4.11. 4. intrinsic rier creams that can decrease permeation of drugs. proliferate on the eschar and prog- the time of the day and year (Hillebrand and ress in depth and make the structure of the barrier Wickett 2008). burn treatment (Moghimi and Manafi 2009). a better understanding of the permeation properties of burnt tissue and its barrier performance 4.1 Intrinsic Barrier Defects towards the permeation of different drugs could lead to improvements in this area. compared to normal ations will help optimised drug delivery and subjects (Seidenari and Giusti 1995). Microorganisms may also colonise the Wickett point out that TEWL can also vary with burn wound. lipid damage. One group is moisturising et al. 2006). cosmetic impaired blood flow and subsequently ischemia ingredients such as surfactants. denaturation of pro- insults. Bullock the summer months. 1980). minimise side effects. environmental or other stressors explained later in this chapter. this review shows that cosmetic condition to become manifest. including exposure to environmental or teins and destruction of protein architecture. While topical therapy is a mainstay of appears to be slower in the aged. as described in the filaggrin (FLG) gene cause a range of SC in previous sections of this chapter. Surfactant. dermatitis characterising this condition (Hachem ous absorption of drugs.12 Compromised Barrier and Its Manipulation Performance Burn injuries to the skin may result in enzyme The SC barrier can be compromised by a range of malfunction.12. day than at night and in the winter rather than in Jackson 1953. physical damage. in Netherton syn. TEWL is higher in the more complicated (Moritz and Henriques 1947. Some surfactants (e. may be involved or required for the inflammatory To conclude. ethanol. It is likely that intrinsic defects in the skin barrier. n-butanol and n-octa- atopic dermatitis in susceptible individuals (Elias nol by up to three times (Behl et al. Similarly. 2008). which may be genetic in origin. including abnor- containing products may also affect percutaneous malities in hydration due to reduced NMF con- absorption. driving the development of to water. where even clinically uninvolved skin other compounds that are possibly applied con. The rate of barrier repair 1996). They and Schmuth 2009). Other cosmetic and ichthyosis vulgaris (Elias and Schmuth products contain ingredients such as glycols or bar. showed that scalding conditions whereby irritant chemicals penetrate with 60 °C water increased the permeability of skin the skin more readily.2 Thermally Damaged Skin 4. Bullock burns. should be considered. TEWL. pH and capacitance. which may cause and necrosis and creation of a dead eschar (Moritz irritation.12. further showed that permeability to all compounds .4 Other Cosmetic Products drome there is a strong association between defective serine protease expression. Generally. their effects on the permeation of dermatitis. including abrasion or and Henriques 1947. causing an There are many other cosmetic products that can abnormally thin SC barrier and the severe atopic affect barrier performance of the skin or percutane. was shown to have defective barrier function by comitantly. Early studies by the Flynn group in the 1980s. 2009). methanol. This may be products are not permeability-inert compounds exemplified by a study in children with atopic and. barrier defects (Hudson 2006). Jackson 1953. Such consider. Thus. and various skin disorders. Monafo and West 1990.g. industrial chemicals. loss-of-function mutations agents that affect hydration level of skin. Reduced filaggrin phate) are well known for their enhancement effects expression has been linked to atopic dermatitis on drug absorption through the skin. tent (Kezic et al. excess water. as barrier defect.

2010). 1981). Organophosphates. Zadeh and at higher temperatures. meate the burn eschar. human third-­degree burn eschar should be noted. While outside the ability was increased by a few fold after heating scope of this review. glycerin. They found that skin perme. Permeation: A Possibility The experiments mentioned above were per. The Moghimi group has also porcine cadavers heated to temperatures ranging achieved success in the use of chemical penetration from 100 to 315 °C.g. Consequently. 2008) as well as terpenes such as orders of magnitude after heating above limonene. not only to prevent permeation of showed that the permeability to methanol and toxic chemicals. the underlying tissues are with a sharp transition in barrier properties occur. therefore. such as amounts to overcome infection and drug delivery parathion and malathion that are used as insecti- to and through burn eschar is still a challenge cides. to 100–150 °C. whereas in the in vivo situation. In vivo experi. continued to increase with increasing temperature. 2008). (Moghimi et al. 1. Grice et al. geraniol. dis. the increase in permeability Moghimi showed that the permeability of human was attributed to morphological and/or chemical third-degree burn eschar was very sensitive to the alterations in the SC (Behl et al. and a Necessity formed on burnt skin separated from the body. the work showing that water. enhancers with burn eschar. as solvents) can reduce et al. permeation enhancement has been the subject of ments also allow the dynamic effects of the burn much research over last few decades. polyethylene glycol) that are ing a maximum on the tenth day. but also due to the fact that some butanol gradually increased after the burn. 2009) were able to increase perme- tively attributed to the disordering of SC lipid ation of silversulfadiazine significantly through structure for temperatures lower than 150 °C. skin responses caused by the burn. followed the progress this. permeable to such a macromolecule (Wang et al.8-cineole and α-pinene oxide 300 °C.68 J. In spite of to be studied. using full-­thickness and increase drug permeation through this barrier skin. ruption of SC keratin network structure at 150– 250 °C and finally decomposition and vaporisation of keratin to create micron-scale 4.g. barrier function Intact skin is a very effective barrier and inhibits is more fully reflective of the range of skin permeation of most compounds. permeability of fully hydrated full-thickness Human skin is exposed to a number of irri- (1500  μm) third-degree burn eschar to silver tants and toxic material in daily life. 1982). while therapeutic substances. For example. nerve agent VX (an organophosphorus ester) . it has been shown that some drugs toxicities. epidermis and SC samples from human and (Zadeh et al. reach. then decreasing employed in dermatological formulations for until termination of the studies at 14 days (Flynn another purposes (e. 2008). Flynn et al. The showed that while fetuin-A (40 kDa) cannot per. 2008. The Moghimi group has investigated percutaneous absorption of drugs (Moghimi and barrier function in humans. there has been interest in effects at lower temperatures were attributed to the techniques to enhance burn eschar permeability to increased hydration associated with scalding. Wang et al. have caused toxicity and deaths after (Moghimi and Manafi 2009). some of sulphadiazine (Zadeh et al. level of hydration and that prolonged exposure to The effects of higher temperatures have been water might open the compact structure of eschar studied recently by Park et al. by one to two orders of magni. These permeability changes were respec.E. which have the ability to permeate the skin bar- Despite the evidence of reduced barrier function rier in toxic amounts and cause local or systemic in burned skin. The 2009). showing increased Shakerinejad 1998). hexane:ethanol and ethyl acetate:ethanol tude after heating to 150–250 °C and by three (Manafi et al.13 P  revention of Percutaneous holes in the SC above 300 °C (Park et al. materials (e. skin penetration retardation is also a very of mice burned with an 80 °C metal surface and crucial issue. also absorption through the skin (Aaron 2001). ring between 70 and 80 °C (Behl et al. particularly if the barrier is damaged cannot permeate the burn eschar in therapeutic (O’Flaherty 2000). 1981).

University of South Australia). Creppy EE (1997) Transplacental 1998). for assistance in editing This area has been extensively reviewed the chapter. as with enhanced SC barrier properties (Pilgram discussed below. which contain ingredients that are sup. Harkness RA. 2008. in prac. mentioned here. effect of topographical polar surface area. the highly hydrophilic polymer. School of Medicine. barrier creams are recommended only for and Dr Amy Holmes. anatomical site.4  Non-formulation Parameters Affecting Skin Permeation 69 shows a percutaneous LD50 of 10 mg for a 70 kg retardants may be explained by their actions to person (Somani and Romano 2000). considerable variability in the skin barrier. pp 819–828 permeation retardation with a range of additives. Camilleri P. 2000). Bonini M. other approaches based on the manipulation of References formulations for preventing or retarding the per- Aaron CK (2001) Organophosphates and carbamates. While most research has creams contain different materials such as surfac. mary protection against high-risk substances. Centre. it has been shown that the Acknowledgements  Thanks go to the National Health percutaneous absorption of trimethylbenzene & Medical Research Council of Australia for financial was increased through skin treated with barrier support. Kilshaw BH. Although beyond the scope of this chapter. Hutt AJ. 2003). Stabilisation of lipid lamellae in the more Different techniques have been developed over ordered orthorhombic phase has been associated time to overcome such transdermal toxicities. 2010). Similar to other creams. The authors also thank Navin Chandrasekaran creams (Korinth et al. Br Med J 2(5964):175–176 . J Chem Inf ation. retardation may be nec- oily phases (that can increase skin hydration). 2012) and polyamidoamine Exp Toxicol 16(9):501–504 (PAMAM) dendrimers (Moghimi et al. WB Saunders Company. et al. (1975) Spongioform myelinopathy in premature taneous permeation seen with certain chemical infants. Hum hydide (Erfan et al. barrier ous skin treatments. 1999). such as polyethylene glycol and Model 52(2):420–428 β-cyclodextrin (Moghimi and Shakerinejad Anane R. Kirton SB (2012) including complexation with high molecular Revisiting the general solubility equation: in silico prediction of aqueous solubility incorporating the weight compounds that show low skin perme. as some gloves cannot prevent pene. care is The interaction of a penetrant with the skin necessary. However. There is creams. the University of Queensland) for help in drawing figures tice. Anderson JM. passage of aluminium from pregnant mice to fetus organs after maternal transcutaneous exposure. For example. for example. (Machado et al. to that under some circumstances. due posed to trap or transform unwanted material or to factors such as the degree of hydration. Kelly RW Kaushik has postulated that the decreased percu. polyan. abnormalities due to ing a protective barrier at the skin surface (Berndt disease or injury and prior treatment with vari- et al. Moghimi et al. gender. so essary in some situations. chemical properties of the molecule or particle 2010). 2010b). have the potential to increase the permeation of drugs. from (Therapeutics Research low-grade irritants and should be not used as pri. cutaneous absorption of chemicals will be briefly In: Clinical toxicology. Ali J. 2010). age. 2010. concentrated on finding ways to increase per- tants (that can act as penetration enhancers) and cutaneous absorption. Another class of protectants is barrier and the properties of the barrier itself. Another increase the organisation of SC lipids (Kaushik example is hexane that can produce a peripheral et al. School of Pharmacy and Medical Sciences. prevent/delay their contact with the skin by form. barrier creams prevent permeation of toxic chemicals. Protective clothing and gloves can provide some degree of protection and are considered to Conclusion be essential protective items. barrier determines its rate of percutaneous tration of low molecular weight chemicals absorption and is influenced by the physico- (Wigger-Alberti and Elsner 1998. Therefore. Brown MB. The Moghimi group has shown Philadelphia. (Therapeutics Research Centre. Kaushik and Michniak-Kohn neurotoxicity through the skin (O’Flaherty 2000).

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91 5. School of Medicine.. SA 5000.1 How Do the Various Emollients Differ in their Ability to Dissolve and Release Different Actives?.....3... Leite-Silva Universidade Federal de São Paulo........ soft). Dragicevic....2..R.............. Shahid Beheshti University Translational Research Institute.... Roberts (*) H..............S............1007/978-3-662-53270-6_5 .5 Emollient Substantivity...1 Sebum ..................... Their Modes C Determined) and to Promote Skin of Action.... University of South Australia....... H..........br the power of softening or relaxing the living ani- J....edu........R....2  urrent Emollients...leite@unifesp. 79 5... 79 Penetration of Actives?...2 Emollient Classes and Properties.....I........ of Medical Sciences. 84 5.....2........... Brazil Dictionary.. 82 5.4 Practical Aspects.. Australia e-mail: hrmoghimi@sbmu....... Australia School of Medicine.. School of Pharmacy Therapeutics Research Centre......2.... The Influence of Emollients on Dermal and Transdermal Drug 5 Delivery V.. Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin...... School of Pharmacy...... DOI 10.ir e-mail: m....2 How Do the Emollients Differ in their Ability to Affect and Enter the Skin 5.. Hamid R...... Instituto de emollire......... 80 Do to Enhance or Inhibit the Effects 5............. Yousuf Mohammed.2.......roberts@uq..3 Effect of Emollients on Skin Lubrication.. SP...... an emollient is a substance “that has e-mail: vania.R..... Moghimi Therapeutics Research Centre.....3  ffects of Emollients E on Percutaneous Absorption..... 77 (Size and Solubility Parameter 5......... 87 and Skin Hydration.. Moghimi.....3 What Do Other Ingredients 5..... out and mol- Ciências Ambientais Químicas e Farmacêuticas...... School of Pharmacy and Medical Sciences......... University of South Australia.” with the first use of the name being Therapeutics Research Centre.......... QLD 4102............. SA 5000...........E. Grice...... Australia e-mail: jeff.........3.....1 Introduction The word “emollient” is derived from the Latin V......... According to the Oxford English Diadema... lis...... and Their Use in Practice.... University of Queensland.......... Jeffrey E........1 Introduction.......... 84 5.grice@uq......... Tehran.2......... 89 5.................... Roberts Contents 5.....3..... and Michael S...2... Translational Research Institute.au M.. 86 5............. Maibach (eds....... Australia Adelaide..........ac.)..................4 Effect of Emollients on Skin TEWL of Emollients on Skin Penetration?.. Iran Woolloongabba........... Leite-Silva........ 80 in a Moisturizing Formulation 5.6 An Overall Comparison of Emollient Conclusion... University of Queensland................... and Medical Sciences.......... QLD 4102.. Grice • Y...........................au © Springer-Verlag Berlin Heidelberg 2017 77 N.edu.. meaning to soften (from ex.. Woolloongabba.. 84 References.. 90 Properties............ 83 5. Mohammed mal textures.... Adelaide.........

and fatty acids. and the patenting of petrolatum hydration) has the sensory feel of softness due to (also known as petroleum jelly. uses. but do not occlude the skin. As he points out. leading to the terms often in nature and are ingredients in a product. Leite-Silva et al. its storage (Marks 2001). stratum corneum (by moisturizing the skin Marks refers to Egyptians and ancient Greeks through reduced transepidermal water loss). Many of the exposed viable epidermis. . and it may contain an emollient and/or a fatty alcohols. although the sensory when applied to the skin deposit a lipid film that responses are different. and tion of the stratum corneum. whereas that about 700 BC.” or to make “vanishing the soothing of the throat (Coxe 1825). white soft paraffin.” “vehicles. its manufacturing. uses. but with. and the access of bacterial. fats and glycerin when applied to the skin tend to soften the epidermis they are termed emollients…” “An adhesive coat is produced which prevents the irritation of drying. emollients are lipid press over the years.R. Interestingly. In general. product. as determined by touch Wehr and Krochmal (1987) 1987 Emollients – systems that smooth the roughened surface of the SC. lanolin is a out the sensorial suppleness feel associated with complex emollient in being a two-phase liquid and the “oily” film.78 V. moisturization of the stratum corneum.1  The nature. and hydration of the earliest emollients were derived from animal fats. which being used interchangeably.1).” the use of wool fat by the Greeks in described as being soft and supple. No effect on TEWL after application Loden (1992) 1992 Similar action to moisturizers Dederen et al. Table 5. and a and that the total lipid content in an emollient for- moisturizer often have become blurred in the mulation is usually 20–40 % (Marks 2001). With a history of several thousands of years. chemical and mechanical irritants” Blank (1957) 1957 Any material that tends to prevent or alleviate the symptoms and signs of dry skin Idson (1982) 1982 Emollients – substances lubricating and/or occluding the skin with water-­insoluble material (Moisturizers – substances actively increasing the water content of the skin) Wilkinson and Moore (1982) 1982 Emolliency is only associated with imparting smoothness and general sense of well-being to the skin. the skin treated with an emollient is on the skin. 2003).” and being lients. A moisturizer usually refers to a wax system consisting of multiple complex sterols. the nature seen as the essential components of emollients” and actions of an emollient. goose the term “emollients” has been interchangeable grease and even human fat have been used as emol- with “moisturizers” and “lubricants.” or “regenerating ing view is that “lipids (fats. the processing of lanolin from treated with a humectant (a substance that attracts sheep’s wool by a Greek physician in a Materia water to the skin (Idson 1992). and oils) are milk” (Nola et al.” “revitalizing creams. nature of the sheep sourced. a soothing effect as it protects the have changed with time (Table 5. using “oils and pleasant smelling fatty concoctions Overall. improving its Medica in 60 AD. Regrettably. waxes. and various definitions of emollients over time Source Date Definition Oxford English Dictionary 1643 Substance “that has the power of softening or relaxing the living animal textures” Edwards (1940) 1940 “Since oils. The resul. and various definitions of emollients skin surface. can also replenish any lost skin lipids. the tant effect is improved skin lubrication. (2012) 2012 Emollients – oily ingredients used for skin care formulations recorded in 1643. a smoother nature. a humectant. In practice. however. A prevail- creams. and Vaseline) in 1872. dependent on the humectant and/or water to provide direct hydra. and emolliency has also been referred to in used as “bases.

emollients affect both hair follicle and consists predominantly of squa- the transepidermal water loss and the roughness lene. (c).1  The effect of various products on the stratum cor. Their vious occlusive films. and transepidermal water loss (TEWL). which emphasized absorption. As shown in Fig. in part. Figure 5. Humectant. Control. An occlusive dressing is also function and the skin penetration of the active. (b). neum. Harry 1941). and some practical examples of prod- the skin hydration associated with emolliency.2.1. ranked in order of increasing lubrication of the stra.2 Current Emollients. Partial mechanical and (ii) the transepidermal water loss (TEWL). Mechanical number of arrows indicating the magnitude of the TEWL Other definitions of emollients have included sepidermal water loss (TEWL) promotes skin the preparations themselves (Ray and Blank hydration. and ointments designed for lient like vegetable oil is likely to reduce TEWL deeper skin penetration (Wild 1911). The ate. (g). 5. occlusion. Confusion to promote skin hydration to a lesser effect than has arisen. often used to increase skin moisturization. ucts containing emollients. of skin surface through the oily film that they cre. and Their hydrocarbon emollients on stratum corneum Use in Practice (SC) roughness. and impairs . 5. with the occlusion with a breathable membrane. (e). (d). compared with other skin treatments such Sebum is the natural emollient of skin. 5. semipermeable or imper. Humectant. sebum provides a pliable film on the skin surface neum and its resulting lubrication of the surface that lubricates the skin. It is evident from this figure 5. we have focused act the symptoms and signs of dryness of the this chapter on the effects of emolliency on skin skin” (Blank 1957). their potential effects on percutaneous advocated by Blank. with a plastic covering. triglycerides. cholesterol esters. In recognizing that the He defined an emollient as “any externally stratum corneum is the main barrier for both der- applied material that tends to prevent or counter. emollient lipid film interactions with skin lipids We now describe the emollients used in prac- and scales and the more mechanistic approach tice. and possibly free cholesterol (Stewart 1992). absorption of unwanted substances. It is pro- as application of humectants and occlusive dress. wax esters. mal and transdermal absorption. the higher the skin hydration) only. The reduction in tran. later times. (f). in 1957.5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 79 Fig. semipolar emollients. early times hydration (the darker the box. and Modes of Action. duced from the sebaceous glands adjacent to the ings. The application of a semipolar emol- 1940.1 summarizes our current view of the effects of humectants. The fig- tum corneum surface after several hours of application ure also shows (i) the effect of products on stratum corneum (except B) for: (a). inhibits percutaneous gives a feel of suppleness. from the early emphasis on the the more occlusive hydrocarbon emollient. Semipolar emollient. The lipid surface film on the stratum cor. hydration.1 Sebum that emollients differ in their actions on normal skin. Hydrocarbon emollient. for example.

3 E  ffect of Emollients on Skin potentially. To the formulator. ple. in a hairless Lubrication mouse model (Lesnik et al. another commonly used humectant. PEG-150 distearate. A plethora of imaginative terms may be restore normal stratum corneum hydration.” “oily. 5. especially when their expression in human skin is phenethyl benzoate. Low sebum levels have been regarded as a meet many different functional needs and to sup- contributing factor in the development of dry skin port multiple “claims. The choice of an emollient is often based on the tum corneum hydration by a glycerol-dependent tactile properties of these substances on the skin mechanism. In addition..2. and lanolin are more remove the sebum and result in greater skin polar than those without these groups. oleyl alcohol. Based on the identification of glyc. and hence. however. ether myristate). and well as measurement of skin hydration (Campbell (iii) an oily feel. among others.g. the nyl isononanoate). gland hypoplasia. ties of the skin in 1977. 1977). cyclomethicone. care ingredients. with articles on the visco- uct. such as Regular washing of the skin. and isopropyl myristate. However. the lipophilicities sebum can also act as a buffer. 1977) and frictional as having a number of common properties: (i) fat (Highley et al. the sebum can also (e. (ii) the ability to soften the skin. soft-draggy 5. Others may feel dry to the touch tion through its surfactant proteins and peptides.” and hence. esters.2) are such that those irritation from acidic or basic compounds. hydrocarbons.g..” “dry-velvety. Today’s emollients are used to tion.2. C12–15 alkyl benzoate. vegetable oils. and isono- upregulated (Mo et al. markedly in their physicochemical properties.2 Emollient Classes feel of raw silk) are less obvious (Goldemberg and Properties and De La Rosa 1971). and are used not only for skin but also for hair tion and hydration of the skin.. impairing adverse of the emollients in (Table 5. a formulator (Clarys and Barel 1995). the tactile sensory proper- Some emollients are partially soluble in water ties of the neat oils are the first consideration in . Urea. containing hydrogen bonding groups. whereas other more esoteric terms such as “scroopy” (the textile chemist’s description of the rough. they can differ quite et al. can ethers. A special issue of the Journal of result in a range of functional and sensorial Investigative Dermatology was devoted to the effects when left as a lipid film after being applied effects of emollients on the mechanical proper- to the skin in a cosmetic or dermatological prod. surface and is often of higher importance in cos- erol as the putative product of triglyceride metology than in the formulation of topical thera- ­hydrolysis in sebaceous glands. PEG/PPG-8/3 laurate) (Stoughton 1959). had no Words such as “tacky. 1977) properties of human skin. where sensory properties are not treated asebia mice. Fluhr et al (2003) peutic drugs. In general. have been shown to enhance sebocyte counts. 2007). Emollients are widely has to select appropriate emollients to meet not used to provide the desired lubrication and skin only the consumer and regulatory needs but also hydration that is normally supported by the to cater for whether the final product is designed sebum. All these terms are used in an attempt to describe the sensory responses Members of the different classes of emollients caused by the lubricating actions of emollients on have different physicochemical properties that the skin. sebum production. leading to increased skin hydration (e. Leite-Silva et al. Sebum has been shown to contribute to stra. used to describe these subjective properties. TEWL.R. PEG-7 glyceryl cocoate. a number of common skin for a cosmetic or a therapeutic application.g. white pet- rolatum. As well as providing lubrica. with glycerol. (e. for exam- roughness and reduce stratum corneum hydra. including mineral oil. Traditionally. “waxy” are readily understood. PPG-3 benzyl provide immunological and antimicrobial protec. emollients have been regarded elastic (Christensen et al. and were able to 2012). showing profound sebaceous necessarily the main priority (Dederen et al. 1992).” or effect.80 V. In addition. as solubility.

5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 81 Table 5.2  Common emollients used in topical formulations Physical form Chemical class INCI name CAS number at 25°C Esters Isopropyl palmitate 142-91-6 Liquid Isopropyl myristate 110-27-0 Liquid Ethylhexyl palmitate 29806-73-3 Liquid Octyl stearate 109-36-4 Liquid Cetyl palmitate 540-10-3 Solid Cetyl lactate 35274-05-6 Semi-solid Myristyl lactate 1323-03-1 Semi-solid C12–15 alkyl benzoate 68411-27-8 Liquid Ethylhexyl isononanoate 71566-49-9 Liquid Isononyl isononanoate 59219-71-5 Liquid Cetyl isononanoate 84878-33-1 Liquid Decyl oleate 3687-46-5 Liquid Diisopropyl adipate 6938-94-9 Liquid Diisobutyl adipate 141-04-8 Liquid Glyceryl stearate 123-94-4 Solid Propylene glycol stearate 1323-39-3 Solid Glycol stearate 31566-31-1 Solid Glycol distearate 627-83-8 Solid Ethers Dicaprylyl ether 629-82-3 Liquid PPG-15 stearyl ether 25231-21-4 Liquid PEG-7 glyceryl cocoate 68201-46-7 Liquid Triglycerides Capric/caprylic triglycerides 65381-09-01 Liquid Fatty alcohols Cetyl alcohol 36653-82-4 Solid Cetearyl alcohol 8005-44-5 Solid Stearyl alcohol 112-92-5 Solid Oleyl alcohol 143-28-2 Liquid Octyldodecanol 34513-50-3 Liquid Fatty acids Oleic acid 112-80-1 Liquid Linoleic acid 60-33-3 Liquid Hydrocarbons Liquid paraffin 8012-95-1/8042-47-5 Liquid Petrolatum 8009-03-8 Solid C9–14 isoparaffin 246538-73-8 Liquid Polyisobutene 9003-27-4 Liquid Isohexadecane 93685-80-4 Liquid Vegetal butters Butyrospermum parkii butter (Shea butter) 194043-92-0 Semi-solid Theobroma cacao seed butter (cocoa butter) 84649-99-0 Semi-solid Mangífera indica seed butter (mango butter) 90063-86-8 Semi-solid Vegetal oils Prunus Amygdalus Dulcis seed oil (sweet 8007-69-0 Liquid almond oil) Vitis vinífera seed oil (grape seed oil) 8024-22-4 Liquid Simmondsia chinensis seed oil (Jojoba oil) 90045-98-0 Liquid Triticum vulgare germ oil (wheat germ oil) 68917-73-7 Liquid Sesamum indicum oil (sesame oil) 8008-74-0 Liquid Esterols Lanolin 8006-54-0 Semi-solid Silicones Cyclopentasiloxane 541-02-6 Liquid Dimethicone 9006-65-9 Liquid Dimethiconol 31692-79-2/70131-­67-­8 Liquid .

Table 5. changes in the However. Accordingly. Spreading values. with spreading isopropyl short-chain alcohol components and the shortest values myristate and isopropyl acid chain lengths (C14 and C16. 2013). measure the mechanical properties of the whole the residual film thickness. In 2013.2. response (Dederen et al. inert surface. but a rigid. alkyl benzo- values benzoate ates. Many attempts have been made to achieve a measure of sensory softness of the skin. evaluation of skin disorders such as atopic derma- This must also contribute to the overall sensory titis (Nakatani et al. Esters are useful to lipid films. resulting in a lower occlusive state ing chain length. 2012). responses to torsional stress. irritation. emollients. This can be assessed quantita. have the highest spreading values. and emollients can directly can also be applied clinically to the quantitative or indirectly modify its mechanical properties. elas- is reflecting is its ability to lubricate and reduce tic responses to ballistic impact. Ethylhexyl values stearate and stearate and decyl oleate. the also facilitates the transport of water through the esters.4 E  ffect of Emollients on Skin by forming a film. TEWL and Skin Hydration tively using a parameter known as the spreading value. example.). so that for lipid films of similar thick- formulators because of their versatility and the ness and viscosity. etc. This important property of emollients is defined by their ability to disperse more or less quickly on the skin surface 5. clothing with skin. were restricted to ronment (skin with skin. paper at 25 °C and measuring the area covered by emollients have been described as indirect skin the applied material in 10 min. choosing an emollient for a cosmetic product Nakatani suggested that conventional methods for (Goldemberg and De La Rosa 1971. They developed a novel piezoelectric tac- sity of the emollient on the skin can be partly tile sensor system that could simultaneously explained by the properties of the emollient itself. In general. skin and its superficial layer. are low spreading esters. A common technique for determining the The residual lipid film on the stratum corneum spreading values has been described by Zeidler surface after the application of products (1985). by dynamic spread. and alcohols.R. includ. have medium spreading values. the semipolar emollients will unique properties they can give to the final prod. C16 palmitate is greasier than the C14 myristate. ­containing emollients will limit the evaporation are determined by applying 20 μl of an emollient of water from the skin surface and therefore to the center of an ashless.3. medium-to-fast filter cause an increase in skin hydration. in units of mm2/10 min. Such a technique has ability and the viscosity. isopropyl myristate and palmitate. be more permeable to water than the hydrocarbon uct. and rheological any friction between the skin surface and its envi. the skin is not obvious advantages to the cosmetic industry.3  Spreading values for selected ester constituent chain lengths can alter the skin-­surface emollients spreading characteristics of ester emollients. influenced by the chemical properties. 1992). and were unable to look at different skin layers sepa- pain (Dederen et al. with longer chain com- decyl oleate ponents. However. measuring the properties of the whole skin and as this reduces possible discomfort. 2012).82 V. are shown in Table 5. respectively) in palmitate this table. of their constituent fatty acids than that induced by the hydrocarbon emollients. these findings can differ significantly. . Zeidler measuring the mechanical properties of the skin. As can be seen. The key property of the emollient that this such as the elongation in response to suction. For High >850 mm2/10 min For example. spreading ethylhexyl but the spreading values are similar. The Medium 501–850 mm2/10 min For example. spreading C12–15 alkyl the longer chain alcohol (C12–C15). spreading values for some of the most widely the presence of hydrogen bonds in emollients used group of emollient for skin lubrication. Examples of moisturizers (Dederen et al. The lubrication inten. rately. 2012). Leite-Silva et al. whereas Low 0–500 mm2/10 min For example.

a simple augmentation of intercellular lipid popu- so that a wax will have a low TEWL and higher lations and structure. The moisturizing effect of water can be nism responsible for a particular skin barrier prolonged when an emollient is present in the defect. relying hydration can be increased by up to 50 % (Hafeez on the use of emollient treatments containing and Maibach 2013a). The occlusive effects of an emollient and isopropyl isostearate cause improved skin then result in a reduced transepidermal water loss hydration and barrier function by stabilizing the (TEWL) and. SC hydration than an oil. showed that to dry skin is to use serine protease inhibitors to the enhanced moisturizing effect of glycerol by treat mild-to-severe barrier abnormalities (Voegeli different emollients and oils was present even 12 et al. The mechanism leading to skin bar- the viscosity of the lipid film will reduce the rier enhancement is believed to involve more than TEWL and increase stratum corneum hydration. 2009) has been described. the applied lipids migrate to the nucleated cell The presence of water in a formulation can add regions. According to associated with skin moisturization was an Elias. such as glycerol. application (Batt and Fairhurst 1986. in turn. free fatty acids. perspiration. and physical occlusives like wound dressings and ceramides at the appropriate physiological pH. et al. 1995. some moisturizers do et al (2012) recently showed that vegetable oils act by penetrating the intercellular lipid regions. but generally for only a short time. imbalance in the SC proteolytic cascade leading ization of the skin. bandages (Voegeli et al. dermatitis or laminar ichthyosis (Pilgram et al. the moisturizing effect of topically augmented lipid mixture is then secreted into the applied water is often lost after 10–20 min of SC intercellular spaces (Mao-­Qiang et al. Increasing has been pioneered by Elias (Chamlin et al. By occluding the skin and pro. Paepe and Chamlin et al. On passing through the SC. This increase in hydration ceramide-dominant physiological mixtures of the as an effect of occlusion has also been seen with three key lipids. such as the action of certain shampoos” (Mosby . 2002.5 Emollient Substantivity being incorporated into the deeper skin layers. and diseases. On the other hand. Patzelt 2001). Rawlings and Voegeli 2013). 2002) to enhance skin barrier func- Rogiers 2009). An alternative approach to address as well as the emollient will increase the moistur. Batt et al. For example. Another definition no effect on the abnormal lipid organization asso. the effectiveness of any such treatment increase in its softness and pliability (Blank depends primarily on understanding the mecha- 1952). 2009. Nonphysiological occlusive moisturizers such as petrolatum remain on the skin surface without 5. where they are on the skin. The presence of a ment is appropriate for that condition (Chamlin humectant. petrolatum treatment had resistance to being rubbed off. Indeed. Blank showed that the main effect tion and normalize skin hydration. 2007). but sion” (Wiechers et al. 2002).2. 2011). 2009. was the most effective where moisturizers such as isostearyl isostearate occlusive. This is defined here as a measure of the retention ing skin hydration. the thickness of the lipid film and/or increasing Elias 2010). A different approach to the emollient treatment viding an additional barrier to water loss. The Indeed. they are not effective in of an emollient in and persistence of its effect on directly treating the disordered lipid states in such the skin after exposure to water. of substantivity is: “the property of continuing ciated with barrier defects in patients with atopic therapeutic action despite removal of the vehicle. orthorhombic phase (Caussin et al. skin of skin diseases such as atopic dermatitis. in the aqueous phase.5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 83 depending on the nature of the emollient. in order to judge whether lipid replace- moisturizing formulation. to be taken up by keratinocytes and then to the moisturizing properties of that formulation trafficked to lamellar bodies. h after application (Batt et al. an increase in the hydration SC lipid organization in the more tightly packed of the stratum corneum relative to normal mois. mixed with endogenous epidermal lipids. cholesterol. petrolatum. ture conditions. 1988). While they may provide some benefit by improv. the semisolid. (except Jojoba oil) led to a similar occlusion of A novel mechanism known as “internal occlu- the human skin surface in vivo as paraffin oil.

2 shows Wiechers’ RPI Stedman’s Medical Dictionary (Stedman’s 2011). 5. A value of ≥70 % for a par- ticular property indicated a good performing An overarching view of percutaneous absorption ingredient. needed for enhanced overall emollient perfor- ter for an O/W emulsion. RPI ≥70 %. bathing. Dermal Torque Meter. The 30 and 70 % cutoffs are shown as dotted tive control and untreated skin as a negative control) and (c) lines (Adapted from Wiechers et al. as silicone tions (Wiechers et al.3. elasticity (using a percutaneous absorption in turn. quantitatively assessing mois. resistance to removal or inactivation by other parameters. each property. did degree of physical and chemical bonding to the not necessarily predict a similar result for the surface. with water applied 30 min under occlusion as a positive control and untreated skin as a negative control). with petrolatum as a posi- range of emollient ingredients shown as individual bars. Figure 5. (MediLexicon 2013). We now consider each of with glycerin as a positive control and untreated the mechanisms by which emollients can affect skin as a negative control). as %) for a Substantivity (using a Sebumeter. effects on the skin. and friction.2  Relative performance indices (RPI.6 A  n Overall Comparison on Percutaneous Absorption of Emollient Properties As discussed in the earlier sections. This led Wiechers to conclude sweating. ­values for moisturization. For (a) Moisturization (by skin hydration using a Corneometer. RPI ≤30 %. referring to performers. among that multiple mechanisms were present (Wiechers other factors.3  ffects of Emollients E 5.2. 2012). Meter. and Release Different Actives? formance index (RPI). he then derived a relative per. RPI between 30 and 70 %.1 H  ow Do the Various (using a Sebumeter. while ingredients with RPI values is that maximal penetration for an active in a sta- between 30 and 70 % were regarded as medium ble formulation will occur at its maximum Fig. so that applied drug or cosmetic. Most recently. medium-­performing ingredi- negative control). impact on the percutaneous absorption of actives turization by skin hydration (using a Corneometer. and substantivity 5. 2002). emollients Wiechers began his pioneering work on skin care can differ greatly in their properties and in their products by working with panels of human sub. Ability to Dissolve For each property. 1997) . in various formulations. swimming. 2009). It is adherent qualities of a sunscreen and its ability to apparent that there are many more medium–good be retained after the skin is exposed to water and performing moisturizing and substantivity ingre- perspiration. determined by the a good RPI for moisturization.” In general. chains are entangled (Sene 2003). low-performing ingredients. tive control and untreated skin as the negative control).R. (b) Elasticity (using a Dermal Torque ents. 5. elasticity. emollient substantivity and Barlow 1999) and that multiple emollients are is poor for an aqueous gel but comparatively bet. Greater substantivity mance (Wiechers et al.84 V. ingredients were classified as: good-perform- with glycerin as a positive control and untreated skin as a ing ingredients. It has been suggested here and in his work to predict the skin penetra- that silicone ingredients have a higher substantiv. with petrolatum as a positive Emollients Differ in their control and untreated skin as the negative c­ ontrol). with water applied 30 min under occlusion as a posi. He tested a broad spectrum of materials affect how these emollients in formulations (Wiechers 1997). Persistence of effect of a topically dients than there are elasticity ingredients. we would be expected for a W/O emulsion and more combined forces to use the principles described particularly for an ointment. Leite-Silva et al. These in turn can greatly jects. tion of actives from complex practical formula- ity than the more common emollients. for example. and substan- suggests substantivity is a “term comprising the tivity for a range of selected emollients.

5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 85 a b c .

as they will not readily . Leite-Silva et al. However.3. for actives of similar size will occur at a lipophilicity similar dissolved in them (Sene 2003). that is. hydrocarbon emol. it will also content in the intercellular regions. that is. The recent concentrations of an active in two emollients will reviews by Hafeez and Maibach examined litera- be higher in the hydrocarbon than in the semipo. but mulation can sometimes result in an enhanced not all compounds (Hafeez and Maibach 2013a). there which would also be expected as the hydrated can be a downside. and structural alterations due to the (Cross et al.2 H  ow Do the Emollients Differ solve an active: “Like dissolves like” and “Oils in their Ability to Affect ain’t all oils”! In other words. 2008). The primary action by which an emollient pro- lients. domains into which the compound must parti- benzone for petrolatum was found to be greater tion. an that occlusion tends to enhance the penetration active formulated with a hydrocarbon emollient of lipophilic compounds more than hydrophilic will usually have a faster rate of skin penetration compounds. However. relatively lipophilic nature of the intercellular Accordingly. 2009). Roberts 2013). a well-recognized strategy for enhancing skin Firsty. promote partitioning from the vehicle into the hydrated skin hydration. and thus substantivity. at saturation. swelling of corneocytes and a rearrangement of These findings have the following implica. tions for how the solubility of an active in an 2010. to that of the skin lipids (a log P of about 3) (Zhang et al.g. Hafeez and Maibach 2013b). lipid-soluble and Enter the Skin (Size actives generally dissolve better in emollients and Solubility Parameter than polar actives. increasing the water content by occlusion lient. Additionally. there appears to be a fall-off in than that for an oil-in-water (o/w) emulsion penetration for very lipophilic compounds. ture data on the effects of occlusion on the pen- lar emollient in accordance with their different etration of compounds of varying lipophilicities relative fractional solubilities. 2012. but will be released stratum corneum. which would be expected. mal. There are two general prin- ciples defining the ability of an emollient to dis. the semipolar emollient will may be further limited. 2001a). for example. with a semipolar emol- skin.86 V. In general. They concluded (Wiechers et al. can lead to enhanced penetration of some. generally deliver its active for a longer period of providing the formulation does not affect the time. Hence. As the active in the hydrocar. skin penetration as a result of the formulation Some possible mechanisms include increased excipients on evaporation. (Kurul and Hekimoglu 2001). the maximal flux for smaller lient. Actives can dissolve in both liquid and motes skin penetration is by hydration of the waxy emollients after heating. the product will be more readily washed off actives is greater than that for larger ones. given the than the one formulated in a semipolar emollient. On the other hand. the thermodynamic activity for two equal penetration (Roberts et al. the intercellular lipid domains (Bjorklund et al. actives dissolve Skin Penetration of Actives? better in semipolar emollients (e. conditions under occlusion increase the water bon emollient has a limited solubility. and in turn skin penetration flux membrane. resulting in a in vivo (Hafeez and Maibach 2013b) and in vitro higher flux of the active through the skin (Hafeez and Maibach 2013a). 5. increasing the viscosity of a for. Silicone emollients appear to offer both those with the lowest melting point (Magnusson persistence in their retention in the skin and a et al. the skin flux for the sunscreen oxy. exhaust much more quickly than in the semipolar the penetration of highly lipophilic compounds emollient. s­ olubility in the formulation. When the skin barrier is nor- more slowly from the waxy than the liquid emol. 2004) and highest flux for a series of actives high solubility. Occlusion is emollient may affect its percutaneous absorption. solubility of the compound in the SC. esters) than in nonpolar. increased solid barrier that may impede TEWL. AND not all emollients have Determined) and to Promote the same properties. and by water. Accordingly.R. leaving behind a semi.. However.

These findings isopropyl adipate (MW 230 Da) is most likely to show an interesting parallel to those of Zhang. maximum flux and lipophilicity for a series of Zhang et al (2013) showed that the ester emol- phenols penetrating human skin in vitro. to carry the phenols into skin lipids. emollients may promote the may strengthen the skin barrier and most likely release of actives from the horny layer (Roberts lead to a reduced permeability to applied chemi. 2004). isopropyl myristate also appeared to domain. partition coefficient of about 3. For example. such emollients. penetration of phenols. domains. including: preservants. Thus. and others. there may be some waxy cals. et al. increasing dent on the relative lipophilicities of the their overall solubility and maximum flux. and concluded that the fin appears not to enter the skin. Therefore. reservoir effect. similar considerations function of the formulation.3 W  hat Do Other Ingredients the skin may have the effect of reducing barrier in a Moisturizing Formulation function and enhancing the penetration of Do to Enhance or Inhibit applied compounds. and extent to which emollients can modify the skin reorganizing the population of intercellular lip. where on Skin Penetration? they become disrupted or fluidized (Kaushik and Michniak-Kohn 2010). retarding the penetration for the As we have seen. if a fra- determine how emollients enter the skin. it may also change . buffers to control the pH. Conversely. This may occur as a result the Effects of Emollients of a direct effect on intercellular lipids.3. In the same way as active solutes will pene. Thus. Results from infrared spectroscopy on emollients which have a very slow release rate human stratum corneum suggested that increased of actives. although functioning as effective a wide range of ingredients with many different moisturizers. a large part of the strategy of more lipophilic phenols. 1992) and nonphysiological lipophilic tivity as a consequence of the emollient’s sub- moisturizers (Caussin et al. leading to reduced rials. emollients. not. A more 2010). isopropyl relationship was driven by the solubility of the myristate is an emollient that enhanced the skin compound in the stratum corneum (Zhang et al. especially emulsions. as shown for silicone esters become localized in separate intercellular (Sene 2003). fragrances. enter the skin. of grance causes skin irritation. and lipophilicity. petrolatum (Ghadially desirable effect is to have an enhanced substan- et al. functions. barrier function (Menon and Ghadially 1997). At the same time. whereas mineral oil did 2009). could potentially hydrogen bonding (Kaushik and Michniak-Kohn cause an enhanced reservoir effect. glycol distea- who showed a parabolic relationship between rate (MW 595 Da) is the least likely to enter. if lipid organization occurred as a result of increased retained in the horny layer. surfactants. The balance between trate the skin according to their size. the application of emollients to 5. were shown by electron micros. the ester emollients in Table 5. with a reduced whereas the hydrocarbon emollient liquid paraf- flux at higher values. thickeners. the liquid di-­ ingly hydrated viable epidermis. stratum cor. melting them is very important for the stability and final point. a restored lipid domain way to occlusion. On the other hand. in a similar ids. with little effect on lamellar organiza- tion or barrier function. 2007) were shown to stantive properties.2. The main effect of isopropyl myristate was neum solubility can be seen to be largely depen. may contain polymers. ­humectants. In both of these examples. chelating agents. Zhang lient isopropyl myristate (MW 270 Da) rapidly saw the greatest penetration at an octanol–water enters the skin and could change its properties. penetrating compound and the intercellular lipid However.5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 87 partition from the stratum corneum to an increas. Certain silicone Formulations. replenishing. However. act as a reservoir. coloring mate- copy to disrupt lipid bilayers. It seems likely that. using emollients to moisturize and soften the skin Limited information is available about the is concerned with replacing. whereas the solid.

et al. usually to solubilize more lipophilic follicles. in order to target the follicular route of actives. 2004). in particular. ability of SDS to penetrate into the SC (Ghosh Nonionic surfactants have also been incorpo- and Blankschtein 2007). alterations in SC bar- tion.g. but can also accentuate emollient effects on ids and bind to keratin filaments (Nokhodchi skin lipids by inhibiting the SC lipid phase transi. mean ± SD) for ten phenolic com- pounds of similar molecular weight from water. isopropyl myristate (IPM). As expected. et al. with the lipophilic sebum environment in hair lations. estimated) versus log P. are capa. they interact at skin penetration by hydrophilic solutes (Wu membrane interfaces and. ingredients in many topical formulations. plotted against log P (Zhang denaturing of skin surface proteins with swelling et al. As they are major d­ isordering of the intercellular lipid structure. 2001). It is clear that the maximum fluxes and disruption of the corneocytes. They act to fluidize lip- tion. mineral oil (MO). with polysorbates being barrier may be significant. in vitro studies using mixtures of ryl sulfate and sodium dodecyl sulfate causing glycerol and the powerful skin irritant and pene. propylene gly. Nonionic surfactants like ethers and poly- showed that glycerol was able to attenuate the sorbates (e. The dotted line interpolates data from the water vehicle and is included as a reference only (Adapted from Zhang et al. human epidermal membranes from a range of tation and skin damage by strong binding and simple vehicles. ble of modifying the structure and properties of Figure 5. anionic and nonionic surfactants. 2013). Anionic and of phenols of similar molecular weight applied to cationic surfactants in particular may cause irri.3 shows maximum fluxes for a series the skin (Williams and Barry 2004). Estimated maximum fluxes (n = 5. in a 1990 study done in a dry atmo. (1990) showed that glycerol enhanced permeation of topically applied mate- acts as a skin moisturizer by inhibiting the lipid rials. skin penetration of actives. with the more rather than by acting primarily as a humectant. and a mixture of 40 % PG/water. plotted against lipophilicity of the phenolic compound. or sorbitol are used to accentuate moisturiza.3  Log (Jmax. disruptive anionic materials such as sodium lau- More recently. rier properties by surfactants may lead to sphere. the Nonionic surfactants tend to cause less irritation effects of humectants and surfactants on the skin and barrier damage. Tween 80) cause more modest effects of SDS on the skin barrier by reducing the degrees of enhancement (Som et al. (Predmore and Li 2011). As the name suggests. accorded GRAS (generally regarded as safe) sta- Humectants such as glycerol. the greatest enhancement (Williams and Barry tration enhancer..al. Most studies have examined the effects of phase transition from liquid to solid crystal. 2003). sodium dodecyl sulfate (SDS). Froebe et. 5.88 V. tus by the US Food and Drug Administration col.R. Indeed. Leite-Silva et al. 2013) . 2012). as well as for water and the occlusive emollient mineral oil Fig. rated into W/O emulsions which are compatible Surfactants are used widely in topical formu.

0 % Cetearyl alcohol 3. 1985.8 % 0.0 % 3.5 Carbomer 0. it may promote skin ing propylene glycol absorption (Zhang et al. hydration than formulation 1.0 % 2.5–6.0 % Triethanolamine pH5. 2001b). emollients to give a smooth and soft feel.4 shows five formulations containing dif- et al. because it lacks oils but is capable where it has been suggested that it integrates into of lowering TEWL ­sufficiently to increase hydra- the stratum corneum lipid matrix and disrupts the tion and potentially promote skin penetration of organization of the lipid lamellae (Brinkmann and an active.5 pH5.5 pH5. skin from vehicles containing propylene glycol Formulation 3 contains mineral oil as an emol- (Grice et al. The trates similar increases in maximum flux seen smoothness derives from the lubrication of the with mixtures of the humectant propylene glycol skin surface by the ester emollient’s lipid film.8 % 0. penetration more. Cross et al. 2010). 2013).0 % 3.0 % 2.0 % 2.8 % 0.15 % 0.5–6.5–6. most likely due to increased solubility this film has a greater effect on TEWL and skin of the phenols in stratum corneum lipids follow. The first impression that a consumer has in using which was due to penetration of IPM into deeper a product is the sensorial feel. The second formulation contains ester Muller-Goymann 2005). as this is Table 5. lient.3 also illus. the enhanced penetration of minoxidil into human and this may inhibit skin penetration of the active.4 Practical Aspects expected for “inert” solvents that do not ­permanently alter the properties of the skin (Barry Table 5.0 % 4.0 % 4.5 pH5.0 % 4.0 % 2.15 % 0.8 % 0. In contrast.5 pH5. the lipid 2011).5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 89 (MO) are similar across this range of log P.8 % parabens (methyl.4  Examples of some formulations containing different emollients and their effects on the skin and the likely skin penetration of an active Raw materials 1 (control) 2 (ester) 3 (mineral oil) 4 (animal fat) 5 (IPM) Ceteareth-20 2.15 % 0.0 % Phenoxyethanol and 0. On the other hand.0 % 4. Formulation 1 feels layers of the stratum corneum (Zhang et al. ethyl. Such a mechanism was also used to explain active is probably more soluble in formulation 2. As and water. particularly for the more polar phenols.15 % Caprylic/capric triglyceride – 5 % – – – (CCT) Lanolin – – – 5 % – Mineral oil – – 5 % – – Isopropyl myristate – – – – 5 % Glycerin 4. and it will feel oily and very soft.5–6. as 5.0 % 3. Figure 5. the ferent emollients and their likely effect on the skin ester emollient IPM led to increased maximum properties and on the skin penetration of an active.0 % 3. fluxes. light and soft.15 % 0.5–6. and propyl) Water Qsp 100 % Qsp 100 % Qsp 100 % Qsp 100 % Qsp 100 % Effects on skin Sensorial feel Light and soft Smooth and Oily and soft Greasy and Smooth and soft heavy soft Reduction in TEWL ↓ ↓ ↓↓↓ ↓↓ ↓↓ Skin hydration ↑ ↑↑ ↑↑↑ ↑↑ ↑↑ Change in solubility of – ↑ ↑ ↑↑ ↑↑ nonpolar active Potential effect on skin ↑ ↑/↑↑ ↑↑↑ ↑↑↑ ↑↑↑ penetration .

This formula. such as the humectant. However. 2012). care must be taken to ensure that these condi. One concern is There are some raw materials that can promote the potential skin irritation caused by these stability of emollient esters at extreme pH. et al. a softens the skin through occlusion and resul- well-known skin penetration enhancer. (Marks 2001). in lipstick). formulations and therefore will provide the great. high pH. tion properties of a product. which may act as skin penetration examination of some emollient-­ containing enhancers. it con. of the active’s low solubility in the mineral oil. They can provide a number of tion enhancement can be achieved (Roberts functions. and ingredients used est inhibition of TEWL and the greatest skin to stabilize emollients in products. Treatments designed to improve dry skin or tancy of the formulations.. may also enhance the skin barrier func- ing lanolin alcohols should be used in formula. propylene glycol (>15 %). Moisturization of the skin is well known There are other practical considerations in the to be the main means by which skin penetra- use of emollients. The most inert the mouth (e. As a general principle. ment. Of the emollients. as it is ingredients must not have an unpleasant taste. treat skin diseases. cholesterol derivatives. emollients may . isopropyl myristate. affect the solubility of an active and penetra- tion of penetration.R. On the other hand. each product has unique physicochem. nition of an emollient. Another The final section discusses some potential out- practical consideration is emollient stability. The overall effect is likely to be an products and their effects on the skin and on enhancement of skin penetration. including the relief of potential dis. However. Leite-Silva et al. skin penetration of an active may be uncertain. 2008). can also cause irritation solubility in formulation 3. tion and reduce its permeability to topical tions in concentrations less than 3 % (Marks chemicals. the most occlusive formulation of all described 2001). among others (Dederen et al. each of which can pH. In very hydrophobic. penetration by inducing a high thermody- ical properties that can promote different interac. Formulation 4 feels greasy. emollients can also comfort and irritation caused by solvents. The key property of an emollient is that it tains the ester emollient. formulation 5 should provide greater lipid film which smooths the skin by lubrica- skin penetration of the active than formulation 2. and explore solubility of the active. it may also promote skin summary. Accordingly. impinge on the sensorial feel and skin penetra- tions will be maintained in a product over time. show the range of tion is expected to be occlusive and to promote the emollients available in the market. ingredients. due to the possibility of hydrolysis or It also needs to be recognized that all cos- saponification.g. this for. and This chapter has attempted to clarify the defi- heavy because of the lanolin waxes. brightness control for makeup. namic activity of the active as a consequence tions and reactions with the skin surface. stabilization tion enhancers. The chapter concludes with an fatty acids. comes for different formulations containing Ester emollient stability may be affected at low or emollients. oleic acid). and free penetration. emollients and other emollient is probably mineral oil and. tion. While formulations metic and pharmaceutical products contain are generally prepared at neutral or slightly acidic different ingredients. lanolin and iso- Additionally. such as lipid replenish- in order to minimize skin irritancy. lanolin also con. An active is also likely to have poorer factants (e. is very similar to formulation 2. promo. Other ingredients. how these emollients are likely to affect skin tains cholesterol. and thus their effects on the of suspensions. Formulation 5 the likely skin penetration of an active. tant moisturization as well as by forming a Accordingly. it should be recognized that if the propyl myristate are the most likely to be asso- finished products are to come into contact with ciated with skin irritation.. the sensitiz. However.90 V. respectively.g. mulation is likely to provide better skin penetra- tion of the active. Another consideration is the potential irri. Conclusion because it contains animal fat such as lanolin. such as sur- hydration. particle coating.

correlation with thermodynamic activity. Carey & Wiechers who was to write this chapter and brought us all Lea. J Invest Dermatol Bjorklund S. Roberts MS (2001b) guidance on the construction of Table 5. We are also grateful to Ricardo Azzini. Pharm Res 18(7):999–1005 Dederen JC. Dugard PH (1985) Vapour and Edwards LD (1940) The role of fats and fat materials in liquid diffusion of model penetrants through human pharmaceutical and cosmetic preparations. understanding the basic relationship between solvent and solute penetration using silicone membranes. hydration effects. factors such as safety. corneum lipid phase transitions in vitro by glycerol – tent of the stratum corneum1. cost. Gooris GS. Harrison SM. Sheu MY. rier perturbation in the presence of glycerol. Friberg SE (1990) Prevention of stratum Blank IH (1952) Factors which influence the water con. Int J Cosmet Sci 8(6):253–264 22(3):245–254 Batt MD. Simion A. instrumentation. based pathogenesis of atopic dermatitis. Gans EH als. Mao-Qiang M. or increasing the solubility of a izers on stratum corneum lipid domains in vitro and chemical or active ingredient in the stratum in vivo. Ghosh S. Probing the effect of vehicles on topical delivery: eral comments. J Cosmet neum.2 and their gen. and the contribu- ing will all be considered by a formulator in tion of the stratum corneum. Clin Dermatol 13(4):307–321 Acknowledgments  This chapter is dedicated to Johann Coxe JR (1825) The American dispensatory. across skin. Hargens CW 3rd. Int J Cosmet Sci 34(6):502–510 Barry BW. sustainability. Oil Soap skin. Glycerol regulates stratum corneum hydration in seba- J Soc Cosmetic Chem 39(6):367–381 ceous gland deficient (asebia) mice. Elias PM (2003) stratum-­corneum following treatment with glycerol. Hadgraft J. Frieden IJ. Blankschtein D (2007) The role of sodium ­isopropyl myristate and a combination of propylene dodecyl sulfate (SDS) micelles in inducing skin bar- glycol and isopropyl myristate on human stratum cor. Pugh WJ. Williams ML. Chavan B. Muller-Goymann CC (2005) An attempt to tion. De La Rosa CP (1971) Correlation of Hydration characteristics and electrical resistivity of skin feel of emollients to their chemical structure. Fairhurst E (1986) Hydration of the stratum cor. Benson HA. Gerrard WA. Fowler AJ. Ann Dermatol neum. Schibli EG. 282–286 Clarys P. Fluhr JW. 17(4):82–84 J Pharm Pharmacol 37(4):226–236 Elias PM (2010) Therapeutic implications of a barrier-­ Batt MD. Davis WB. Rawlings AV (2012) Emollients References are more than sensory ingredients: the case of isostea- ryl isostearate.5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 91 enhance the barrier’s permeability by mecha. stratum corneum using a noninvasive four-point J Soc Cosmet Chem 22(10):635–654 . 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Ghafourian T. Roberts MS. Medical Dictionary. Pirot F. Vissers DC. Leite-Silva et al. Int J Cosmet Sci 23(4):211–218 Predmore A. Cross SE. ple. Voegeli R (2013) Stratum corneum prote- Loden M (1992) The increase in skin hydration after ases and dry skin conditions. Int J Pharm percutaneous penetration of chemicals in man and 250(2):359–369 monkey: partition coefficient effects. Barzegar-Jalali M (2003) The tive review. Medical Terminology. Stuttgart. Shokri J. Weiner N. Bhatia K. Nokhodchi A. Lavrijsen SP. Richter H. Elsevier – Stewart ME (1992) Sebaceous gland lipids. Roberts Nakatani M. Kawasoe T. Skin Res Technol Hafeez F. Cell Tissue Res 351(2): application of emollients with different amounts of 217–235 lipids.92 V. Blank IH (1940) Effect of ointment bases on the Magnusson BM. Results of patch tests with commonly used oint- (2004) Molecular size as the main determinant of sol. J Invest Dermatol 127(2): the health professions and nursing. Divergent mechanisms for correction of affecting the formation of a skin reservoir for topi- permeability barrier dysfunction. Arch Derm Syphilol 42(2):285–289 ute maximum flux across the skin. Sene C (2003) Silicone excipients for aesthetically supe- Menon G. Kankavi O. Kostovic K. From http://www. and bath oils. Hekimoglu S (2001) Skin permeation of two dif. Cross SE. AAPS PharmSciTech 11(3):1068–1083 Aberrant lipid organization in stratum corneum of Kurul E. van der Meulen H. Br J Dermatol Syphilis Acta Dermatovenerol Croatica ADC 11(2):80–87 53(3):65–82 Paepe K. Informa Healthcare. 303–305 pp 279–294 Idson B (1982) Moisturizers. Microsc Res Tech tions.

Barlow T (1999) Skin moisturisation and Zeidler U (1992) Über die taktilen Eigenschaften kosme- elasticity originate from at least two different mecha. Int J Pharm 220(1–2): Wiechers J (1997) Relative performance testing: introduc. Dederen JC. Joller P. Br J Dermatol 161(1):70–77 Wilkinson JB (ed) Harry’s cosmeticology. Krochmal L (1987) Considerations in selecting Topical transport of hydrophilic compounds using a moisturizer. DC Wild RB (1911) On the official ointments. Watkinson AC. Informa Zhang Q. Roberts MS Wiechers JW. New York. tischer Öle. Li P. Doppler S. Jepps OG. Int J Cosmet Sci 21(6):425–435 Zhang Q. Cosmet Toiletr 112(9):79–84 Fette Seifen Anstrichmittel 87(10):403–408 Wiechers JM. Adv Drug Deliv Rev 56(5):603–618 atous atopic skin. Inc. Roessler BJ (2001) Wehr RF. In: Rothman formulation and inter active effects during formulation S (ed) The human integument. Li P. reference to the substances used as bases. p 62 Rawlings AV (2011) Increased mass levels of certain Williams AC. Starmans WA (2009) Skin solubility determines maximum transepi- (2002) Multifunctionality: from “One in More” to dermal flux for similar size molecules. Li P. J 1911:161–162 neum serine protease activity in acute eczematous Wilkinson JB. Fluhr JW. In: Rawlings Release 154(1):50–57 AV. Zeidler U (1985) On the spreading of lipids on the skin. Rawlings AV. Barry BW (2004) Penetration enhancers. Rawlings AV (2009) Zhang Q. Leyden JJ (eds) Skin moisturization. optimization for transdermal delivery. Weiner ND. Wortel VAL. Relation of the anatomy of normal complex cosmetic formulations: an evaluation of inter and abnormal skin to its protective function. Int J Cosmet Sci American Association for the Advancement of Science. Cross SE. 63–75 ing a tool to facilitate cosmetic ingredient selection. New York. Wang GJ. Ramachandran C. J Control nism of action of skin moisturization. Roberts MS (2011) Maximum transepider- Moisturization mechanisms: internal occlusion by mal flux for similar size phenolic compounds is orthorhombic lipid phase stabilizers-a novel mecha. In: atopic skin. Roberts MS size phenolic compounds. Grice JE. Pharm Res 30(1):32–40 (2012) Predicting skin penetration of actives from . SÖFW 118:1001–1007 nisms. Int J Cosmet Sci 33(6):560–565 Wu H. Doppler S. Chemical Voegeli R. Normal and abnormal. Br Med Schreier T. Breternitz M. Verboom C. Fluhr JW (2009) Increased stratum cor.5  The Influence of Emollients on Dermal and Transdermal Drug Delivery 93 Stoughton RB (1959). 34(6):525–535 Washington. serine proteases in the stratum corneum in acute eczem. Moore RJ (1982). Liu D. enhanced by solvent uptake into the skin. Pharm Res “More in One”. Roberts MS (2013) Effect of vehi- Healthcare USA. Cosmet Toiletr 117(4):73–78 26(8):1974–1985 Wiechers JW. Cutis 39(6):512–515 water-in-oil nanoemulsions. Breternitz M. Skin creams. pp 309–321 cles on the maximum transepidermal flux of similar Wiechers JW. with special Voegeli R. Publishing.

the composition of the 6.3 Potential Mechanisms of Interaction.............. 1987.. Tsuruta 1996..... Kansas State University..... Jepson and J...... College of Veterinary Medicine. USA Karande and Mitragotri 2009).... 97 substances usually occurs with the substance car- 6... 99 solvents used in extraction processes. Chittenden and Jim E....... North Carolina State University.... 103 properties of the penetrant..... Cross et al. 1986.. 2007... 99 pharmaceutical applications.......T............................ 99 by-products...... 2001..... H.......... Traynor 5520 Dillard Drive........1 Skin Surface.3........... 113 The impact of different vehicles and enhanc- ers on percutaneous absorption has been studied extensively (Carpentieri-Rodrigues et al...........chittenden@certara..... McDougal 1999..... A large number J..... The ability to predict 6..2.... 96 In industrial settings. Williams and Barry 2004)..3 Epidermis and Dermis..... 102 used to stabilize the formulation and modulate absorption.....2 Partition Coefficient...................... In either case....... NC 27518.. et al..................... USA et al...... surfactants............1 Solubility....2 Thermodynamics of Mixtures......... Manhattan....I..... Dragicevic............. For 6.. KS 66506-5802.............4... ATS of studies have demonstrated synergistic effects Coles 207.....3 Diffusion... References... 104 the effect of vehicle composition on the relevant Predicting Absorption from Complex 6................... Walker and Smith e-mail: jason. Dupuis et al........ 95 6. The mixture may contain 6.2 Stratum Corneum...... Riviere Contents 6.... wetting agents...4 Experimental Assessment of Interactions.............. The Effects of Vehicle Mixtures on Transdermal Absorption: 6 Thermodynamics.... 2006...1 Introduction 6.... 2007..edu have shown permeability enhancement in binary © Springer-Verlag Berlin Heidelberg 2017 95 N...........com 1996.. PhD... DSc(hon)... Sherertz et al... 103 6...1 Introduction..2 Isolated Perfused Porcine Skin Flap.. de la Maza et al.............. dermal exposure to toxic 6...........2. 113 the risk in toxic exposure or maximizing the ther- apeutic value in pharmaceutical applications...... 2007............E...2... Mills Pharsight Corporation.. Riviere........................... Assessment.........3. 1992......... reaction 6............. Mechanisms.......... Maibach (eds........... 103 vehicle affects the thermodynamic and transport 6.......... 104 the penetrant would be valuable for evaluating Conclusion..... DVM...... 1998.1 Diffusion Cells. and Prediction Jason T.....5  properties of the penetrant and model exposure to Vehicle Mixtures... a dosing vehicle is 6. Cary...... etc. 2007........3...4........................... suite 260...... enhancers (Carpentieri-Rodrigues et al............. 98 ried in a liquid mixture............ Mills 2007.. Dias et al........... Van der Raleigh.. of combinations of solvents and/or penetration Institute of Computational Comparative Medicine..... 2005.4......... 101 6.... NC...... USA Merwe and Riviere 2005a. DOI 10.........3 Model Membrane Systems............. Several studies e-mail: jriviere@ksu.. Chittenden (*) Biomathematics........ Department of Anatomy and Physiology.... Hotchkiss et al....1007/978-3-662-53270-6_6 .).... Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin....

Van strengths and limitations of available data. are not suit. 2002) of quantitative differential change in Gibbs energy of a system structure to permeability relationships (QSPRs) with respect to one of its constituents. Single penetrant or where R is the ideal gas constant. reviewed to highlight the mixture effects that can affect some key properties driving permeability. Despite such a erty of a penetrant. 2010). text of potential mechanisms for interaction with The activity coefficient of component i can then vehicle constituents. Chittenden and J. the chemical potential is the definitive showed only one example where a vehicle effect property which is equated in various equilibria was accounted for. have been developed. Monti et al. with the best results on average for ternary absorption can be related back to a single prop- combinations (Arora et al.. T and P are enhancer studies. experimental apparatuses gives insight into the Baynes et al. Finally. enhancers. models to predict vehicle mixture effects requires è ¶N i øT . and it only provided an indi. One study explored over 4000 binary vehicles to develop rules for predict. equation leads to an expression of the change in The goal of this chapter is to exhibit some of the partial Gibbs energy of mixing (ΔGi) as a the most recent works in the modeling and pre. aqueous or single vehicles. and in nonstandardized conditions. even if combined. with the concentration of added surfactants (Shokri et al. An overview of some of the be taken as: . (e.mi o = RT Inxi + RT In g i underlie many of the interactions are briefly = DGi ID + DGi EX .T. phase. 2003). der Merwe and Riviere 2005b. The chemical thermodynamics that DG i = mi . An extensive study of up to quaternary combinations of ten enhancers on The key properties – solubility. which large body of work. a comprehensive is a function of the composition of the multicom- ­understanding of the effect of complex vehicle ponent system also including solvents and mixtures is still elusive. mi is the chemical potential is only recently that predictive models that of pure component i at a reference state ( T  . the chemical potential. For compo- cator variable to adapt between two vehicles. xi is the mole fraction able to study synergistic effects.g. 6.2 Thermodynamics ing synergistic permeability enhancements of Mixtures (Karande et al. El Maghraby et al. In practical for percutaneous permeation prior to 2002 terms. reaction. partition coeffi- skin permeability found synergistic enhance. temperature and pressure. Building æ ¶G ö mi = ç ÷ = RT ln g i xi + mi . include vehicle effects. adsorption). cient. 2001). excess Gibbs mixing energies for component i. one can write the chemical Most of the prior work on skin absorption is done potential μi as with few compounds. Enhancement of diazepam ture on predicting mixture effects on permeabil- permeability in rat skin from a 50:50 mixture of ity shows the promise and failings of work to propylene glycol and water was found to vary date.E. Studies should of component i. Nj are the numbers of molecules be specifically designed for the purpose. and diffusivity – that drive transdermal ment. and especially mixture and γi is the activity coefficient. a thorough review of the available litera- Rosado et al. Thus. it of each component j. nent i in a solution. P . P  ). where ΔGiID and ΔGiEX are the ideal and The structure of the skin is discussed in the con. 1995. The chemical potential measures the A review (Moss et al. 2009.96 J. function of concentration: diction of skin permeability from complex vehi- cle mixtures. 2001. 2006). Rearranging the effects. Riviere or ternary mixtures (Baynes and Riviere 2004. N j ¹i an extensive dataset with dosing applied in vari- ous vehicle compositions.

6  The Effects of Vehicle Mixtures on Transdermal Absorption 97 DGiEX Mirmehrabi proposed a simple activity coef- = ln g i ficient model. Many are incremental improve.1). and diffusion. Activity coefficient models. A simple one-parameter Wilson model has been used to predict the solubility of four drugs in water plus cosolvents of either ethanol. chemical property predictions. of Matsuda et al.i æ ö T Chen and Song (2004) by introducing the con- ln xiL = ÷ . been used to model 14 drug compounds’ solubil- ble physical properties. Application of the NRTL-SAC model to vehicle Many activity coefficient models have been mixtures showed good agreement with experi- developed and applied to solubility in drug-like mental data (Chen and Crafts 2006). even in compounds. UNIQUAC and NRTL outper- way to include mixture composition effects in formed in mixture systems (Mirmehrabi et al. These range from simple mod. similar to the Margules model but RT extended to mixtures.4-dioxane (Matsuda et al. Functional Activity Coefficients) (Mirmehrabi nism to link that departure to the mixture compo. xiL is the solubility. and polarity. L = xiS g iS f i . xiL g iL f i . provide a binary systems.ln g i ç1 - L cept of a segment activity coefficient (SAC) to RT è Tm ø create the NRTL-SAC model. The NRTL-SAC model has prediction of solubility can be based on accessi. The bility of stearic acid in a solvent mixture with activity coefficient captures the departure from better results than UNIFAQ (UNIQUAC ideality for real mixtures and provides a mecha. The polymer g f i . S The basic NRTL model was successfully If the solid is pure. coefficient of salicylic acid in water and ethanol. extend its capabilities to associating systems .1 Solubility methanol. and used it to predict Chemical potential is the driving force behind vapor-liquid equilibria in 41 systems and solu- solubility. et al. ous vehicles is important both in manufacturing performed with the one-parameter Wilson model processes and in the formulation of drug prod. and els to complex computational chemistry-based captured the solubility peak at minimum simulations. 6. So. phase equilibrium. or 1. vents showing that while it performed well for tions between mixture components. (2010). phase activity. 2010). The solubility of a compound in a solvent is change in activity due to variation of the mixture the equilibrium expressed as: composition from pure water to pure ethanol (Fig. Computations of the infinite dilution activity Predicting the solubility of a compound in vari. then this reduced to applied to correlate simvastatin solubility in 15 solvents. 2006a). L i L NRTL model of Chen (1993) was extended by DH m . ments on already existing models such as An adapted version of the UNIFAC model. activity. The NRTL-SAC where ΔHm. S are the based on the segments’ hydrophilicity. Tm is the model decomposes molecules into recognized melting point. S xiL = mixtures (Nti-Gyabaah et al. 6. associating systems such as ethanol/water. and f i . which tested with three drugs in pure and mixed sol- incorporate parameters that describe the interac. and a model for liquid ity in up to 14 solvents (Chen and Song 2004). hydropho- fugacity of the pure liquid and solid phases.2. L and f i . was developed by Mengarelli to UNIversal QUAsiChemical (UNIQUAC). highlight the large ucts. γiL is the liquid segments and adjusts the activity prediction activity at saturation.i is the enthalpy of fusion. the ­ Non-Random Two-Liquid (NRTL) and A-UNIFAC. 2006b). of the solute. The same proposed model was sition. 2009). bicity. with the goal of predicting solubility in f i .

The tures (Spyriouni et al. in which NRTL-SAC RT ln g ia xia + mi = RT ln g ib xib + mi and the modified UNIFAC fared best (Diedrichs xia g ia = xib g ib and Gmehling 2010). The NRTL and for modeling vapor-liquid and liquid-liquid sys. 2012). though the NRTL-SAC model gave a minimum.0 mole fraction Ethanol (Mengarelli et al. A com. The equilibrium between two liquid parison of UNIFAC.0 parameter (Wilson model with parameters from Matsuda et al. and an ab initio method (Tung et al. where Cα and Cβ are the concentration of solute in tures. The partition coefficient is the ratio of concentra- pounds in up to 28 pure solvents and was found to tions of a solute in two different liquid phases. 2010) 10. 1999).2 0. C a xia va g ib va sion of COSMO and NRTL-­SAC for four drugs in P= = = C b xib v b g ia v b 13 solvents and found that while NRTL-SAC pre- dicted better overall. 6. equilibrium. Conductor-like Screening phases α and β at a constant temperature (T) and MOdel for Realistic Solvents (COSMO-RS). and pranolol in a water/n-octanol system. pressure (P) is governed by: NRTL-SAC.0 acid in water and ethanol mixture. SPARC performs automated reasoning in . The A-UNIFAC model 6.2. and a pharma-­ modified UNIFAC mia = mib was made. temperature (Mohsen-Nia et al.5 0. Gross and Sadowski developed the perturbed The equilibrium concentrations are determined chain statistical associating fluid theory (PC-SAFT) by the activity coefficients (or vice versa to com- (Gross and Sadowski 2001) as an equation of state pute activity from observed data). UNIQUAC? models have been used to correlate tems. It was extended to associating systems such partition coefficient data for atenolol and pro- as ethanol/water (Gross and Sadowski 2002). Chittenden and J. COSMO-SAC had the advantage of being the two phases.8 1. using up to 17 drugs in 15 pure sol- vents and a few mixtures. Riviere Fig.0 0. 2008).0 1. vα and vβ are the molar densities of the two phases. At be adequate.2 Partition Coefficient was extended and compared with NRTL-SAC in predicting solubility for seven drug-like com.1 Activity coefficient of salicylic 100.E. 2011).98 J.0 g0 salicylic acid 5. this represents a ternary system in generally better results (Mota et al. P is the partition coefficient. Tung compared a SAC ver.T. including in solvent mix. 2012). by one 50.4 0.0 0. with both has been applied to the prediction of solubility of showing the ability to predict P as a function of six drugs in up to 17 pure solvents and several mix.6 0.

RMS = 0. RMS = 0. If the has been computed from observed Fickian diffu. where N is the number of compounds D12 = ( D12 ) (D ) x1  x2 in the dataset and RMS is the root mean square 12 . 2004). The vehicle can then have synergistic effects by acting in multiple lev- In a binary system (or perhaps one with only one els. The results showed that D could of a neutral ­solute (penetrant) in a pure solvent.487). and penetration of drug from factor Γ.3 Diffusion 6. activity tration of the solute and that the Maxwellian dif- coefficient (N = 2647. The thermodynamic Transport of the penetrant from the dosing vehicle factor can be computed from any activity coeffi.272). RMS = . Several mechanisms at the skin’s surface affect the fusion coefficient D12 and the thermodynamic dissolution. 2011). then dissolution sion coefficients D for several binary systems may also become a rate-limiting factor.3 Potential Mechanisms Chemical potential plays a role in diffusion as of Interaction well. where D12 and D12 are the error in the prediction in log units. it encounters different = åGij Ñx j j =1 environments at each level. however. Note that the mixture molar density ct is the dosing vehicle into the stratum corneum. How these properties n æ x J -x J ö di = å ç i j j i ÷÷ interact. being described by: et al. into RT ç j =1 è ¶x j ø solution at the skin surface. vary by an order of magnitude with the concen- dict solubility (N = 707. then into the stratum n -1 corneum and dermis. solubility of the penetrant in the dosing vehicle or The activity-adjusted diffusion coefficient D12 the rate of diffusion across a boundary layer.1 Skin Surface experimental data is the Fickian diffusion coeffi- cient D. the entire range of concentrations. Ingram used infinite dilution diffusivities.44) (Hilal solute was log-linear. As the penetrant moves x ¶ ln g i ö di = i Ñmi = å ç d ij + xi ÷÷ Ñx j from the dosing vehicle (if there is one). The implication is COSMO-RS in the prediction of partition and that with an activity coefficient model for a sol- distribution coefficients of 19 ionizable drugs vent/solvent system. diffusion. 6. the . at different levels of the pene- ç j =1 è c D tration process is the key driver of the rate and t ij ø n -1 æ extent of absorption. In addition.6  The Effects of Vehicle Mixtures on Transdermal Absorption 99 c­ hemistry (SPARC) system has been used to pre. fusivity D12 as a function of concentration of tion coefficient (N = 698. and distribu. the equation simplifies to: composition of the solvent and/or absorption environment can have dynamic effects on the J1 = -ct D12 GÑx1 = -ct DÑx1 chemical properties related to penetration and æ ¶ ln g 1 ö absorption.3. taken to be constant here. to the surface of the skin may be limited by the cient model. mechanisms that change the diffusing solute).2. D = D12 G = D12 ç1 + x1 ÷ è ¶x1 ø The diffusion coefficient usually estimated from 6. The generalized Maxwell-Stefan equation relates the diffusional flux of components of a Thermodynamics provides a mechanism for mixture to the driving force for diffusion (Taylor understanding how components in a mixture can and Kooijman 1991): interact to modulate key properties related to transdermal penetration. In the case (Vignes 1966). that diffusivity can be predicted over (Ingram et al. penetrant is dosed as a suspension. and a prediction of dilute over a range of pH and with various counterions diffusivity. which is the product of the Maxwell dif.

2012b). D is slower flux and crystallization of penetrant on the diffusion coefficient. Solvation imposes increased may be increased. Supersaturation of lipophilic solutes in the rounding surfactant molecules have an increased donor well has been shown to increase their tendency to move as a unit. If the bulk concentra. . For binary or higher as the effective volume of solvent decreases mixtures of solvents. When surfactant concentrations are above possibly above) the saturation concentration the critical micelle concentration (CMC). which molecules instantaneously when they disassem. centration. where J is flux of ¶z tum corneum. even if not ideal (in the sense of “ideal” cause an increase in solute concentration thermodynamic behavior). impact on absorption is more complex. Nonetheless. however. reduces rate of diffusion. occluded and evaporation could not occur. etration into the skin. evaporation can ­simple. loss of solvent is through its pen- 1989). In the case of incomplete evaporation. presuming no other order around the solute molecule. rapid intial flux will give way to penetrant.100 J. it may begin to pre- boundary layer at steady state is determined by cipitate. This activity while not affecting the stratum cor- implies a trade-off between increased solubility neum barrier function (Moser et al. however. Micelles diffuse much solves. composition over time (except in the case of an Consider a boundary layer of the dosing vehi. If the solvent evaporates completely ¶C before the solute is fully absorbed into the stra- Fick’s law as: J = . may be overall slower than if the system were ble at the stratum corneum interface. azeotrope).g. The flux of penetrant through the due to solvent evaporation. with the end result of incom- the boundary layer. a drug partition coefficient. can deliver multiple solute lowering of flux to the dissolution rate. the combined (Stinchcomb et al. is that the solvent does not evaporate com- tion of penetrant is near saturation. and effects on solubility or diffusion coefficient. One other possibility not in SC membrane itself). rate-limiting step. until all the precipitated drug redis- plicating the system.. Riviere thermodynamics of the system are relatively If volatile solvents are used. the point of precipitation. As the solute concentrates eluting patch). If. the One way to increase solubility is to add sur. which is more likely in a solvent mix- ture effects that increase solubility can increase ture where one of the components is not flux as long as diffusion to the skin surface is the volatile. and some bulk fluid or dosing apparatus (e. z is the distance through the skin surface.D ( Ch . instead of penetration is generally reduced (Wiechers evaporation. In essence. increasing flux and reduced diffusivity reducing When the evaporation stops. phase Csat and should remain at the saturation con- separation into micelles will occur. then the mix. how- J = . Up to factants. Assuming a constant gradient plete absorption (Oliveira et al. These same effects may occur if. the evaporation of one systems. C is concentration of penetrant. 1999). the penetration by increasing thermodynamic effective radius of the solute increases. as the solute and sur. Ch will be at (or flux. but may be an initial increase in flux followed by due to their size. further com.T.C0 ) / h . the effect will be to increase penetrant concentration at the skin surface (note. and as such may exhibit synergistic of the solvents results in a change in the solvent effects. the absorption rate phobic molecules. If redissolution is slow. When combinations of the properties in the pure solvent a solvent mixture is used. the solvent evaporates more slowly than ­concentration at the top of the layer and C0 is the the absorption rate. 2001). Chittenden and J. diffusivity. which can aid in the solvation of hydro. pletely.E. with the introduction of micelles. where Ch is the bulk ever.D . which also increases flux. which can drastically change the cle sandwiched between the skin surface and physical properties of solubility. the net effect more slowly than the single solute molecule. This disappearance of effects on the solute properties such as solubility solvent volume can also reduce the depth of the and diffusion coefficient are not necessarily simple boundary layer. the overall absorption rate.

The effective. arrangement of the corneocytes in the “brick and Disruptions to the packing structure in the mortar” model of the stratum corneum creates a crystalline regime or introduction of moieties tortuous path that effectively increases the length that displace the small chain lipids and choles- of the lipid pathway by about an order of magni.3. or loosening marily neutral lipids and sphingolipids (largely of the keratin structure in corneocytes (Daniels ceramides) (Lampe et al. corneocyte lipid envelops. lamellae. partition coefficient between the vehicle and stra. 1989) The transit and accumulation of a penetrant with various ceramides. layer and increasing fluidity. displacement of water domains. Bouwstra 1997). 2001). free fatty acids. terol in the fluid phase can weaken the diffusional tude (factor of 12. with the lipid head lar matrix of lipid lamellae. 1991). It anisms by which vehicle constituents could affect has been determined that the presence of endog- the stratum corneum barrier function are extrac. with phase between them with embedded head groups the lipid lamellae contributing the majority of of the shorter chain lipids and cholesterol filling that function (Squier et al. and any mol. 1991). cent layers meet. As previously stated.6  The Effects of Vehicle Mixtures on Transdermal Absorption 101 6. and the presence of free fatty amplifies the impact of the mixture. lipid and cholesterol sulfate. which is generally oriented parallel to the resulting in their extraction. The tight pack- by the total lipid content of the stratum corneum ing of the head groups in the crystalline regime (Grubauer et al. 2012. In addition. groups oriented at the corneocyte membranes. though some liquid phase components to influence one or both of them may be present. The barrier function on both sides where the lipid head groups of adja- of the skin is primarily due to the stratum cor. can extract lipids from the intercellular . the diffusional barrier is tum corneum lipids. The lipids are organized primarily of these phenomena and the potential for mixture in crystalline phases. The lamellae are stacked multiple times over ness of the barrier to water transit is modulated to fill in the intercellular pathway. 2003). 2003). corneocyte envelops. The “brick acids results in a denser packing structure and mortar” model of the stratum corneum (Bouwstra et al. The mech. ponents may completely disrupt the lipid layers. The and the orderliness of the fluid regime provide a only continuous pathway through the stratum strong barrier to diffusion of large and hydro- corneum is via the lipid multilayer. 1983) with some polar and Knie 2007). and cho- through the stratum corneum is governed primar. 1996). orthorhombic packing structure. as pathway for diffusion is the intracellular lipid opposed to the hexagonal or liquid-crystal phases. The lateral diffusion of lipo- ecules diffusing through the stratum corneum philic solutes in the stratum corneum lipids is must traverse the lipid multilayer or move within dependent on the weight and size of the molecule it (Bouwstra 1997). enzymes. Squier et al. the overlapping (Johnson et al. 1989. Wertz et al. Many permeation-­ stratum corneum surface (Bouwstra 1997). philic compounds. alteration of the tion (Bouwstra et al. 1975) suggests that the main denser. disruption of the lipid bilay. 1987. 1975. and struc. enhancing compounds. in. The presence of the (Michaels et al. Some vehicle com- The lipid lamellae are arranged in a multi. layer. The interaction between them. These lipids are The stratum corneum is composed of flattened formed into bilayers that span the gap between corneocytes stacked in layers with an intracellu. These are pri- ers. The lamellar layers consist of a crystalline regime tural proteins (Elias 2012). lesterol (Bouwstra 1997) comprising the lamellae ily by partitioning and diffusion. the intercellular lamellae of lipids. which provide a hydrophobic barrier to correlates with the effectiveness of the stratum water loss from the underlying tissue. enous proteins does not affect the lipid organiza- tion of lipids from the lamellae. and a more disordered fluid neum (Baroni et al. Talreja et al. such as alcohols and A lipid monolayer is chemically bonded to the alkanes.2 Stratum Corneum k­ eratinized cell membranes of the corneocytes (Swartzendruber et al. corneum barrier (Damien and Boncheva 2009). corneoctyes in several layers.7 in human skin samples) barrier by disrupting the ordered structure of the (Michaels et al.

(Ibrahim and Li 2010). Increasing the partition- within the solvent fraction/solvent-rich areas ing into the cell membranes or the fluidity of the inside the membrane or skin by increasing solu. various that the penetrant is eliminated by desquamation. and possible that small polar molecules could follow ­propylene glycol demonstrated that partitioning an aqueous paracellular pathway through tight into the stratum corneum was determined by the junctions to the basal lamina (Brandner 2009. ceramides (Vávrová et al. enhancers that increase blood flow would create a ple. Depending upon the desorption rate. possibly and Smith 1996). though a titioning into the stratum corneum (Hilton et al. 1995). vehicle components that raise γsc or As the penetrant moves through the epidermis. Metabolizing enzymes in the epidermis may Conversely. Inclusion of water in the stra- zation (Kitagawa et al. That is. was found to sharply increase the diffusiv- occur by modification of the mixture properties ity of water in the membrane at values over 80 % on the skin surface or through changes induced RH (Wiechers 1989). (Pieper et al. 1990). if desorption is so slow et al. (Stinchcomb 2003. Systemic uptake is a depot effect of the drug. increasing . as a function of relative humidity Modulation of the partition coefficient can (RH). Remaining compo- actions where high solvent uptake promotes nents can interact in a few interesting ways to drug partitioning by enabling the solute to exist enhance absorption rate. For exam. Another potential interaction in the stratum cor.3. modula- other compounds including ethers and alcohols tion of this accumulation may be crucial. Fatty acids (Aungst 1989). co-penetrant that inhibits or downregulates expres- 1994). Suhonen or even reduced absorption. lowering the solubility in the vehicle reduce overall bioavailability of the penetrant has been found to also contribute to greater par. A study of the effect cells (Monteiro-Riviere 2010). between the tissue and plasma. 2012a). sion of the enzymes will diminish that effect.E. which is a function of the C0 v0 x0g 0 6. Riviere lipid domains of the stratum corneum (Walker corneum increased for some compounds. 2011). which may cause a ability of the penetrant. it is of vehicle mixtures of SLS. 2003). 2003). it lower γ0 of the penetrant increase the partitioning must cross cell membranes and diffuse through into the stratum corneum. partition coefficient in delipidized stratum stronger sink condition for diffusion. By the time the pene- stratum corneum lipids (Van der Merwe and trant arrives in the epidermis. Experiments in model mem. At high concentrations. and numerous terpenes Hydration of the stratum corneum results in (Williams and Barry 1991) have been shown to the inclusion of bulk water that may pool in cor- increase permeation by fluidization of the lipid neocytes or within the intercellular lipid matrix lamellae. barrier between the skin and the systemic avail- tinocytes (Frasch et al. resulting in disruption of the crystal structure and this could result in an extended time of absorption increased permeability (Krill et al.3 Epidermis and Dermis mixture composition through the activity coeffi- cients. These results may indicate in the stratum corneum lipid structure. Acidic pH may also contribute to fluidi. while at lower is normally shielded by the lipid domain (Surber concentrations it disrupts the lipid head groups et al.102 J. Chittenden and J. tum corneum. 1999). Storm et al. the vasculature acts as the final neum is binding of the drug to proteins in the kera. it is likely to be sepa- Riviere 2005b). 1992. rated from the hydrophilic penetration enhancers branes and skin support a model of vehicle inter. relative solubility of the solute in the vehicle and O’Neill and Garrod 2011). Alternatively. ethanol. The parti. ethanol due to increased exposure to a protein domain that has been shown to extract lipids. lipids and the vehicle at the skin surface is C v x g Psc = sc = sc sc sc . when the tion coefficient between the stratum corneum stratum corneum is in a highly hydrated state. 1990). penetrant may function of the perfusion rate and the partitioning move rapidly into the stratum corneum but accumu. membranes will increase diffusion of the pene- bility in the membrane (Oliveira et al. a variety of For penetrants with local action or toxicity. In the dermis. So. trant. and other vehicle components. the formation of a hydrophilic pathway.T. water. Penetration late there rather than enter the epidermis.

fresh perfusate is continually intro- many tissues in the body. Finally. which indi. while in the flow- venous uptake of the penetrant. duced and collected (Bronaugh and Stewart eral drug transporter systems. Co-penetrants that affect protein ents) partition into and diffuse through the binding of the penetrant in the extracellular membrane to the receptor well. The P-glycoprotein 1985. For penetrants. binding. and AUC (area under the Skin Flap concentration profile) which indicates extent of exposure. and may not provide much informa. unethical (in the case of ity to assess effects due to perfusion or metabo- toxicants). by stirring or turbu- from the systemic circulation. collection of data over several penetrants fused with a modified Krebs-Ringer solution and vehicle mixture combinations is required and containing glucose. and cytokine signaling above the fixated membrane sample is dosed (Riviere et al. In trying to identify tion studies. such as solubility. the skin expresses sev. then the rate-limit- P-gp may provide a boost to penetration rate that ing barrier to permeation will be diffusion co-penetrants could inhibit (Skazik et al. The perfus- ate effluent can be collected or sampled for analysis to determine the absorptive flux of pen- 6. lence. and only feasible in a high-throughput system. serum albumin to maintain viability. antimicrobial agents. integrated tion on the mechanism of the interaction. The IPPSF is surgically prepared in molecular properties of vehicle constituents that situ on weanling swine and excised after closure contribute to changes in penetrant transport prop. 6. The use of different membranes such as silas- tic and porcine skin allows the combination of 6. In vitro tissues. .4. inflammation. Franz 1975). it is common (P-gp) transporter system is one of the most to include a binding agent such as albumin in the important in presenting barriers to drug uptake by perfusate to create a sink condition on the recep- tissues. But. whether static or flow-through. The integral. Riviere and Monteiro- with a penetrant and vehicle mixture. In vivo experiments ble or otherwise accounted for – diffusivity. the receptor well is a closed system the equilibration into the blood and the eventual which is periodically sampled.4. lism which only manifest in viable. If the boundary layer in the donor and vasculature and presents a barrier to drug uptake receptor wells is kept small. of the incisions. through the membrane. A donor well lism. The isolated perfused porcine skin flap and ex vivo experiments provide a way to isolate (IPPSF) addresses these deficiencies by provid- mechanisms of interaction as well as increase the ing an ex vivo system for percutaneous penetra- throughput of experiments. In both cases. in vivo experiments can be expen.6  The Effects of Vehicle Mixtures on Transdermal Absorption 103 permeation rate (Riviere and Williams 1992. metabo- ­determining skin permeation rate. investigation of effects on permeation and parti- presents a conceptually simple system for tioning due to changes in perfusion. or if diffusion is rapid. like through cell. 2011).1 Diffusion Cells etrants placed on the surface of the flap. P-gp is concentrated at the tor side. offer the most direct assessment of effects via endpoints such as Cmax (maximum plasma con- centration) and Tmax (time of Cmax). Riviere 1991). viable nature of the IPPSF allows the Diffusion cells. time consuming. trant (and perhaps some of the vehicle constitu- Wiechers 1989). and if the penetrant parti- the transport properties of a penetrant can be tioning/binding to silicone membrane is negligi- assessed in several ways. The flap is cannulated and per- erties. In the Franz-type spaces of the dermis or blood plasma will affect static cells. In the dermis.2 I solated Perfused Porcine cate rate of exposure.4 Experimental Assessment the obtained data to factor out the contributions of Interactions from the membrane and account for effects in the donor and receptor wells. The interaction of vehicle constituents to modify evaporation. The pene. A deficiency with in vitro systems is the inabil- sive. 1986.

material on a 96-well microtiter plate and placed in a 96-well receptor plate containing buffer. and used to understand diffusion and partition.3 Model Membrane Systems of vehicles on dermal absorption has been recently reported (Karadzovska and Riviere Model membrane systems have been developed 2013). Silicone-­MCF potential correlations to transport properties. The fibers are exposed to the donor encountered with in vitro and in vivo stratum solution for a fixed incubation time. The major limitation of this method is the tinal permeability (Avdeef 2012). and from Complex Vehicle flux is measured after a suitable incubation time. Samples taken well before equilibrium can ever. Wertz et al. Suhonen et al. PA-MCF. to regress data and obtain partition coefficient pholipids into SC lipid liposomes increases flu. how. These systems array. The Parallel been applied to the study of vehicle mixture inter- Artificial Membrane Permeability Assay actions (Baynes et al. Partitioning the idea of using multiple MCFs in an array has of absorption enhancers into SC lipid liposomes led to a unique assay for transdermal permeation has been correlated to increased permeability using PDMS-MCF. Another high-throughput technique for perme- ing of compounds as well as the effects of vehi. Half of the wells are used with the membrane and half are used as controls (Kansy et al.4.5 Predicting Absorption solutions are introduced on the donor side. and polyacrylate (PA-MCF) membranes. while at equilib- changes induced by vehicle components and rium they will assess partitioning. ability assay is the membrane-coated fiber (MCF) cles on membrane structure. partition coefficient correlated well ( R 2 = 0.93 ) in 32 compounds (Xia structure is an effective approach for enabling et al. descriptive mathematical model that can be used 2008. PAMPA con. The composite estradiol. 1999). (PAMPA) has been used for years to assess intes. 1986). Extending solubilization (López et al. A study mate partitioning and absorption of jet fuel com- of the interaction of sodium dodecyl sulfate ponents in polydimethylsiloxane (PDMS-MCF) with SC lipid-like liposomes found that increas. progesterone. 2005). They do. need for study compounds to be amenable to sists of membrane constituents fixed on a filter assay by gas chromatography. 2007). such as diffu- . Chittenden and J. 2010). 2004). The multiple MCF approach has also high-throughput experiments. wherein model membranes are immobi- lack the complex structure and interactions lized on fibers. allow for a finer focus on the structural be used to assess permeability. Riviere 6. Mixtures Recent developments have led to a modified PAMPA system that shows promise for predict. 1995. estimates when equilibration time is inconve- idity of the membranes and partitioning of niently long (Xia et al. 2003). ship (QSPR) is a mathematical equation that The use of a PAMPA system to predict the effects relates a property of permeability.T. showing ing cholesterol sulfate content in the membrane excellent correlations between partitioning and increased the resistance of the membrane to observed log Ko : w (Xia et al. 2012).E. MS. 2007.104 J. and propranolol into approach (MCF with regression) was used to esti- the liposomes (Kirjavainen et al. and Wax-MCF through fluidization of the membrane (Ibrahim (CarboWaxcoated) partition coefficients jointly to and Li 2010). A quantitative structure to permeability relation- ing transdermal permeability (Sinkó et al. 2008. 1998). the entire fiber can be processed by GC/ ies and metabolic interactions. Incorporation of phos. Riviere et al. after which corneum. predict porcine skin diffusion cell log Kp with Fixation of model membranes to an ancillary high fidelity ( R 2 = 0. Permeant 6.927 ) Lipids extracted from the stratum corneum with the octanol/water partition coefficient (Xia (SC) can be studied as liposomes or lamellar et al. The MCF technique is bolstered by a sheets (Kitagawa et al. 2000). such as corneocyte envelop boundar.

ϕv(n) is the volume fraction of solvent n size is sufficient to describe stratum corneum (which approaches unity for dilute solutions). to a linear function of chemical descrip- sivity would be approximately constant. eth. Using a coefficients.(d i .6  The Effects of Vehicle Mixtures on Transdermal Absorption 105 sion or partition coefficients. resulting in a permanent Abraham and Martins performed a metastudy change of barrier function – lipid extraction. However. Sloan reasoned of interest. includ- ing dimethylformamide. especially in the case of . it failed molecular size (volume) and Koct is a surrogate to predict much higher fluxes from isopropyl for SC lipid partitioning. coefficient cients to skin permeability coefficient worked MW . etc. They note that ionization of the mechanism for predicting changes in barrier compounds does not necessarily have a deleteri- function due to vehicle application. These are models that relate a property (Barton 1975.d v ( n ) Vifv2( n ) / RT v n and Martins 2004).1) well for theophylline in various vehicles. diffu. that aqueous pathways play a significant role in Substituting into the definition of partition the absorption kinetics in SC (Potts and Guy coefficient: 1992). LFERs have been used to relate various therefore the permeability rate constant kp would molecular properties to overall permeability depend only on partition coefficient. and tors.3RT K oct . for involving experiments on human skin for 119 instance. Extrapolation across exper- imental systems should not be expected. ous effect on absorption. This first widely myristate and octanol vehicles. ln Pi v ( 2 .1) solubility parameters of the solute i and vehicle that accounts for lipid partitioning and molecular n. accepted QSAR for predicting skin permeability lar solubility parameters to skin. predicting Kp has arisen from their use in pre- ute i in solution n: dicting various other physicochemical proper- ties such as log partition coefficient (Abraham ( ) 2 ln g i ( ) = d i . it ­vehicle) such as molecular weight. gives: 0 compound Vi ( = æç d i .d v ( n ) . which in turn is energy relations (LFER) (Karadzovska et al. 1986). dipolar. and formamide. may be of limited use in practical application. molecular weight is a surrogate for ylene glycol. propylene glycol. δi and δv(n) are the Potts and Guy found that a simple model (6. Sloan et al. to chemical tional problem with this approach is that it relies descriptors of the permeant (and possibly on saturation conditions in the donor well.67 ) for the 93 compounds in the Flynn vehicle that changed the diffusivity or chemical dataset (Flynn 1990). which has its basis in Gibbs free that for small molecules of a similar size.b ¢MW 0 D .diffusion path length 2 s .1) sat = ln Xi ( v 2 ) sat Xi ( ) = ln g iv (1) sat ( d ) + f log ( K log ( k p ) = log D 0 oct ) v 1 sat g iv ( 2) sat . and T is temperature.hypothetical diffusivity of MW = for vehicle n and skin s. An early relationship that could be Another approach to the incorporation of ­considered a QSPR related partition coefficient activity coefficients is through linear free in skin to solubility parameters. where V is the molar volume. nature of the skin. so.octonal : water partition The correlation of the predicted partition coeffi. v ( n ) sat 2 log Pi è ø 2.1).Molecular weight (6. ity. In (6. R permeability. This might have coefficient showed a strong correlation been due to an interaction between the skin and ( R 2 = 0. this approach does not provide a compounds. and they discount competing ideas is the gas contant. a method for computing activity coefficients 2013a). So. The use of LFER equations for relationship for the activity coefficient of the sol. An addi.d s ) ö÷ ) d . Ko : w. which have simi. energy.

and 20  % water. lar size (Hostynek and Magee 1997). but the approach lacks general sp . because it uses an indicator variable E . A similar approach was also used to study benzidine absorption in IPPSF log ( k p ) = i + aVEH +bMR + cHBA+ dHBD (Baynes et al. with the MDCMs com- Hostynek and Magee proposed a model posed of a solvent (acetone or.106 ( ±0. culated for four compounds over 13 vehicles. observed flux in IPPSF. (1996) used the MDCM data of Accounting for the vehicle effects on the perme. Assessment of absorption and residue levels indi- tional feature of their model is a factor for dosing cated not only effects due to single vehicles. and their uptake of the compound must necessarily include concentrations in the various compartments are some factor to account for the presence of the allowed to affect the rate parameters in mecha- dosing vehicle. involving molar refractivity (MR). One addi. R = 0.excess molar volume that ignores the relative concentrations of the vehicle and compound. compounds will occur parathion that compared quite well to the in the presence of solubilizers and/or permeabil. (1996) proposed a 2 mechanistically defined chemical mixture N = 119. selectively a surfactant.461. surfactant (sodium hydrogen bond acceptors (HBA). They fit a model of system rather than separated out as features of the form: interest (Moss et al. nicotinic acid (MNA)).E. metabolism modifier (stan- hydrogen bond donors (HBD). number of dimethyl sulfoxide (DMSO)). in most of the literature up to nistically chosen. or dosing of.log ( k p ) or log ( k sc ) applicability. The MDCM is defined as containing a penetrant.åhydrogen bond basicity pretreating skin samples with various vehicles V . and The positive term for volume seems nonintuitive. ing for all terms except characteristic volume: While most of the studied vehicles enhanced par- titioning into SC. Hostynek and Magee (1997) provide a simple model for accounting for sp = c + eE + sS + aA+bB + vV the vehicle effect. The effects of 16 MDCMs on para- ular volume and hydrophilicity (Geinoz et al.152 ) vehicle mixtures on the permeability of a particu- B + 2.473 ( ±0. a vasodilator. In addition. nearby compartments. 2 × 4 factorial design. Qiao et al. solvent(s).129 ) coefficients of the permeants. full name.McGowan characteristic volume before applying permeants in a diffusion cell. SD = 0.148 ) A − 3. each of the ity enhancers.000 ( ±0.296 ( ±0.426 ( ±0. Chittenden and J. a vasoactive compound (methyl accounts for effects of lipophilicity and molecu. DMSO in the SC increases the . (1996) to predict and compare trans- ability constant is an important aspect in trans. This model also nous chloride). a reducing agent.137 )V lar compound. these important ing to a modified Hill equation.832. and number of lauryl sulfate (SLS)).T. F = 112 (MDCM) experimental design. Yet.101) − 0.106 J. and/or but is due to negative correlation between molec. for aspects were integrated into the experimental instance. That is. a few modulated diffusion log ( k p ) = −5. (2003) examined the effect of B . dermal flux and skin residues using a multicom- dermal absorption modeling. E − . accord- the turn of the 21st century. Riviere hydrogen-basic molecules.dipolarity / polarizability ship is not extensible to new vehicle effects. the accurate prediction of the components of the MDCM is modeled. In this model. 1996).473 ( ±0. but vehicle (VEH): also interaction effects. Flux and retention enhancement ratios were cal- The final fitted model for kp shows it is decreas. the relation- S . Williams et al. A .095 ) S To separate and model the effect of complex −0. Qiao et al. thion absorption in IPPSF were evaluated in a full 2002). Because most partment dermato-pharmacokinetic model for exposure to.åhydrogen bond acidity Rosado et al. 2002).

6  The Effects of Vehicle Mixtures on Transdermal Absorption 107

p­ ermeability of parathion, but does not affect its treatments with four to five replicates per treat-
transfer to the depot compartment. Parameter ment. A validation dataset with five compounds
values for vehicle transit were directly gathered (also included triazine) in various chemical mix-
from previous studies, while those for parathion tures added 56 additional treatments. Finite doses
were adjusted from a previous study. While this were applied which resulted in dynamic flux pro-
model is certainly mechanistic and thorough, it files; so, permeability constant (kp) was assessed
requires many parameters for each penetrant, and from the regressed slope of the cumulative flux
many more for the interactions, making estima- profiles.
tion and validations of such a large model imprac- Among the 16 LFER relationships reviewed
tical for prospective applications. by Geinoz et al. (2004), the training set of 288
Extending the experimental approach of the treatments was fit well by Abraham’s model,
MDCMs, Riviere and Brooks have found signifi- which was used as a baseline fit for investigating
cant improvements in LFER fits, using the potential mixture factors. Potential MFs were
Abraham and Martins models to describe perme- identified by trends in residual values against MF
ability in porcine skin flow-through (PSFT) cells values. Three MFs identified as having potential
by accounting for mixture effects with a mixture to reduce the residual variance (based on R2 of
factor (MF) (Riviere and Brooks 2005). The MF the trend) were refractive index, polarizability,
approach adds a new term to the LFER that is a and log(1/HC).Inclusion of these MFs in the
mass fraction weighted mixture property of the LFER improved the correlation as shown in
penetrant-vehicle mixture. The basic form of the Table 6.1. Predictions on the validation set data
mixture factor (MF) is as follows: showed that the MF approach extended well to
the new treatment combinations as well.
MF = å mi pi
chemcial = i
It is worth noting that the descriptors used in
mi - mass fraction of chemicali the MFs have a link back to chemical thermody-
namics that make their effectiveness less than
pi - property of chhemicali surprising. Consider that Abraham’s LFER
These experiments examined 20 chemical prop- accounts for diffusivity and partitioning compo-
erties to assess the influence of their MF on nents of kp and that the primary role of the MF is
­permeability, including descriptors of molecular to account for a change in partitioning due to the
size, volume, boiling point, hydrogen bonding vehicle. The partition coefficient is a function of
properties, polarizability, refractivity, melting the activity coefficients of the solute in the vehi-
point (MP), and more. Twelve chemically diverse cle and skin, but the LFER is optimized for an
penetrants and 24 vehicle mixtures (comprised of aqueous vehicle. Correcting a partition coeffi-
water, ethanol, propylene glycol, sodium lauryl cient for the vehicle composition, then, would
sulfate, and methyl nicotinate) were studied in require multiplying by the ratio of activity coef-
PSFT in a full factorial design resulting in 288 ficients in vehicle and water. Henry’s law con-

Table 6.1  Coefficient of determination (R2), of predicted vs. observed log kp and regression residuals vs. mixture factor
(MF) value
Original dataset (288 treatments) Validation dataset (56 treatments)
Predicted vs. Predicted vs.
Mixture factor observed log kp Residuals vs. MF observed log kp Residuals vs. MF
None 0.58 0 0.62 0
Refractive index 0.80 0.53 0.78 0.41
Polarizability 0.76 0.43 0.69 0.17
log(1/HC) 0.77 0.45 0.75 0.34
Table modified from Riviere and Brooks (2005)
HC Henry’s law constant, log Kp log of permeability coefficient, MF mixture factor

108 J.T. Chittenden and J.E. Riviere

stant is the activity coefficient of an infinitely d­ ifferent experimental systems to extrapolate sin-
dilute solute; so, it has direct bearing on the com- gle-compound LFER models across different dos-
putation. Polarizability relates to the attractive ing vehicles, although the specific chemical
forces between molecules in solution, which property that best describes the vehicle effect has
have direct bearing on activity. Refractive index yet to be identified. An additional study in PSFTs
itself is influenced by polarizability with a posi- using a different set of compounds (caffeine, cor-
tive correlation. The MFs, however, are an imper- tisone, diclofenac, mannitol, salicylic acid, and
fect measure of the required activity coefficient testosterone) was recently reported where the
ratio due to the incomplete information provided optimal MF was 1/MP for these drugs
by these descriptors and the potentially inade- (Karadzovska et al. 2013b). This study also
quate linear mass fraction mixing rule used. allowed the incorporation of datasets from differ-
Nonetheless, the success with this approach high- ent in vitro models (infinite versus finite dosing,
lights the promise and necessity of incorporating saturation level, vehicle) to be accomplished by
the vehicle mixture properties when predicting incorporating indicator variables and a MF of 1/
skin permeability. MP.
A further study used the same data from PSFT A QSAR model developed with the already
cells (Riviere and Brooks 2005) and additional published data (Riviere and Brooks 2005) again
data for ten penetrants in five vehicle mixtures in highlighted the importance of vehicle effects by
IPPSF to investigate the effect of adding mixture finding a significant correlation of kp with the dif-
factors to the LFERs of Potts and Guy, Hostyneck ference between melting and boiling point of the
and Magee, and Abraham and Martins (Riviere vehicle (Ghafourian et al. 2010a). Stepwise
and Brooks 2007). The mixture factor approach regression with descriptors for the penetrants
was able to improve the kp prediction in each of including connectivity indices, quantum molecu-
the LFERs tested in PSFT. The results from the lar properties, and group counts, along with vehi-
Abraham and Martins based model are shown in cle mixture properties comprising melting point,
Fig. 6.2. Note the vertical banding in (a) due to boiling point, solubility, vapor pressure, and
the penetrant predictions being unadjusted for Henry’s law constant, resulted in a best fit model
mixture effects, as well as the large variation in which included octanol/water partition coeffi-
observed log kp values. In many cases, the vari- cient, ninth order molecular connectivity index,
ance within a penetrant, due to the formulation and difference in boiling and melting point of the
effects, is larger than the variance between pene- solvent system.
trants. Inclusion of topical polar surface area
log k p = −0.909 − 0.610 log K o:w + 2.62 c p ,9
(TPSA) as a MF allows the predictions to vary
within a penetrant, removing the banding effect, −0.00917 ( BPs − MPs )
(6.2)
and reducing the prediction error. Application of
MFs to predict area under the flux curve (AUC) In the resulting model, Eq. (6.2), Ko : w is the octanol-­
in IPPSF also improved the prediction over the water partition coefficient, χp,9 is the ninth order
unmodified LFER models. molecular connectivity index, and BPs and MPs are
Influential MFs in PSFT were number of the boiling and melting points of the solvent. The
hydrogen bond acceptors, refractive index, and negative effect of partition coefficient was surpris-
TPSA. TPSA provided the best fit among all of ing, and it was hypothesized that this was due to the
the investigated LFERs. For IPPSF, octanol:water overall lipophilic nature of the dataset. This moti-
partition coefficient and inverse water solubility vated the author to compare the chemical space of
were important MFs for the Potts and Guy LFER, the dataset with the datasets of Flynn (1990) and
while log water solubility and ovality (a measure Wilschut et al. (1995). The comparison was con-
of deviation from spherical shape) were important ducted using principal component analysis, where
in the other two LFERs investigated. The results linear combinations of the descriptors are used to
showed that the MF approach works in two describe the variability of the response variable.

6  The Effects of Vehicle Mixtures on Transdermal Absorption 109

a

b

Fig. 6.2  Predicted vs. observed of penetrant absorption topical polar surface area (Figure from (Riviere and
in PSFT cells, with the Abraham and Martins LFER Brooks 2007), used with permission)
(Abraham and Martins 2004). (a) With no MF. (b) MF is

Plotting the first two principal components against With the data for the four additional chemicals
each other (Fig. 6.3) is enlightening as it demon- expanding the dataset to 384 treatments and
strates that the chemical space of the dataset is increasing the diversity of the chemical space, a
quite constrained when compared to the chemical QSAR study was again performed using stepwise
space of the Flynn and Wilschut datasets. selection across a wide array of chemical

110 J.T. Chittenden and J.E. Riviere

Fig. 6.3 The chemical diversity of the combined datasets in PCA analysis (Figure modified from (Ghafourian et al.
(Riviere – solid circles, Flynn (1990) and Wilschut et al. 2010a), reprinted with permission. *PC1 and PC2 are the
(1995) – empty circles) using all of the chemical descriptors first and second principal components of the chemical space)

d­escriptors and physicochemical properties of
the penetrant and solvent mixture (Ghafourian higher boiling points will result in lower absorp-
et al. 2010b). Solvent mixture properties were tion. The Wiener topological index, which is a
­computed by mass weighted average, similar to measure of molecular size, accounts for reduc-
the MF approach. The model size was con- tion in permeability with increasing molecular
strained to no more than four descriptors to avoid volume. The negative contribution of lipole may
spurious correlations. The model (6.3) with the reflect the fact that this descriptor combines
best fit ( R 2 = 0.701 ) included descriptors for: the lipophilicity,which is usually seen to increase
difference in melting point between the ­vehicle permeability, with molecular size, which should
and penetrant ( Dmp = mpv - mp p ); Wiener’s decrease permeability. Testosterone, the largest
topological index of the penetrant (Wp); boiling molecule by Wp, showed the greatest variation
point of the vehicle (BPv); and lipole of the pen- in permeability due to vehicle composition and
etrant (Lp). was an outlier in some of the treatments, which
indicates that the QSAR is not able to account
log k p = -0.956 - 0.00322Dmp fully for vehicle effects in some of the
-0.000320W p mixtures.
-0.0121BPv - 0.114 L p A study that combined data from PSFT (16
(6.3)
penetrants in 384 treatments) and IPPSF experi-
The implications of this model are that pene- ments (20 penetrants in 119 treatments) resulted
trants with lower melting points will have higher in a total set of 27 penetrants in vehicles com-
absorption rates. In addition, vehicles with posed of mixtures of water, methylnicotinic acid,

6  The Effects of Vehicle Mixtures on Transdermal Absorption 111

sodium lauryl sulfate, propylene glycol, and eth- absorption. The IPPSF includes dermis, vascula-
anol (Riviere and Brooks 2011). A dataset with ture, active enzymes, and is a live tissue – all of
seven compounds in mixtures of 14 vehicle com- which can contribute to not only differences in bar-
ponents (89 treatments) in IPPSF was used for rier function, but also to tissue partitioning and
validation. QSPR models based on the LFERs of depot effects. In addition, the MF approach appears
Potts and Guy, and Abraham and Martins, were to confound effects in the solvent with effects in the
estimated separately for the two experimental barrier, such that for the same vehicle mixture dif-
systems. A stepwise selection process was ferent MFs are required. It is suggested that using
applied to the mixture factor components added the estimation from PSFT to fix some portion of
to the LFER models, resulting in the final QSPRs the model for IPPSF may help to isolate effects that
(6.4) and (6.5). are particular to the ex vivo IPPSF system.
A QSPR for the absorption of cosmetic and
log Y = i + mMF + a åa 2H + båb 2H
dermatological formulations in human skin was
+ sp 2H + eR2 + vVx (6.4) developed from a model for predicting cumula-
tive absorption (Bunge et al. 1995; Bunge and
log Y = i + mMF + a log K o:w + bMW (6.5)
Cleek 1995; Cleek and Bunge 1993), adjusting
In Eqs. (6.4) and (6.5), Ψ is a placeholder for it to account for ionization of the penetrant in
either kp (in the case of PSFT) or AUC (in the the vehicle as well as distribution to aqueous
case of IPPSF); MF is a placeholder for a specific and organic phases of an emulsion (Grégoire
et al. 2009). The adjustment for ionization
mixture factor; åa H
2
is hydrogen bond donor affects the prediction of partitioning into skin
acidity; åb H
2
is hydrogen bond acceptor basic- and is accomplished by replacing the log Po : w
term in the Potts and Guy LFER with the log
ity; π2H is dipolarity/polarizability; R2 is excess
distribution coefficient of the penetrant in the
molar refractivity; Vx is McGowan volume; Ko : w
octanol-water system at the pH of the vehicle
is octanol:water partition coefficient; MW is
(log DpH). The effective area of diffusion was
molecular weight; and m, a, b, s, e, v, and i are
corrected for the aqueous fraction, which is
estimated parameters. In both the PSFT and
assumed to be the phase contributing the major-
IPPSF datasets, the MFs that contributed most to
ity of the flux. The model assumes steady-state
improving the model, as evaluated by correlation
diffusion, ignores the effect of penetration
coefficients and cross-validation metrics, were
enhancers, approximates all vehicles as emul-
the number of hydrogen bond acceptors (HBA)
sions, and considers only absorption from the
and the inverse of melting point (1/MP). However,
aqueous phase of the vehicle.
different MFs were needed in each model system
The dataset comprised cumulative amount of
to achieve the best fit. The QSPR based on the
absorbed measurements from human skin in 101
model of Abraham and Martins could be simpli-
individual studies, with a total of 36 compounds
fied by removing a parameter, but it was a differ-
in various vehicles and experimental settings.
ent parameter in each of the model systems:
The estimated parameter of the model is a coef-
hydrogen bond donor basicity in PSFT and polar-
ficient to control the distribution of the penetrant
izability in IPPSF. The addition of a MF to a
between the phases of the emulsion. The result-
LFER model has little impact on the estimated
ing QSPR predicted 91 % of the data within 0.2
parameters (Table 6.2) for the chemical descrip-
tors, which indicates the independence of the MF to 5-fold ( R 2 = 0.7115 ). This model is unique in
from the descriptors. the literature in incorporating biphasic vehicle
The results would indicate that different barrier and ionization interactions, but more research
functions are being evaluated between the two sys- needs to be done to extend it to ionic or metabo-
tems, but they could also indicate a ­fundamental lized compounds, monophasic vehicles, and to
difference between predicting rate and extent of account for penetration-enhancing excipients.

112 J.T. Chittenden and J.E. Riviere

Table 6.2  Compilation of QSPR results from Riviere and Brooks (2011), showing best fits with and without mixture
factor (MF)
R2 System Equation MF m i a b s e v
0.53 PSFT (6.4) – – 2.55 1.45 0.01 0.27 −0.55 −1.39
0.73 PSFT (6.4) HBA −1.21 4.27 1.44 – 0.28 −0.55 −1.38
0.60 IPPSF (6.4) – – 1.61 −2.00 −0.39 0.04 0.38 −1.54
0.69 IPPSF (6.4) 1/MP 35.63 1.81 −2.00 −0.35 – 0.47 −1.61
0.33 PSFT (6.5) – – 1.13 −0.10 −0.0058
0.55 PSFT (6.5) HBA −1.23 2.89 −0.10 −0.0058
HBA hydrogen bond acceptors, MP melting point, PSFT porcine skin flow-through cell, IPPSF isolated perfused por-
cine skin flap, m, i, a, b, s, e, v, which are parameters in Eqs. 6.4 or 6.5, as indicated by the equation column (NB
Parameters and carry different meanings in the two equations)

The effects of vehicle properties and experi- and melting point ( BP - MP ( mix ) ), showing
mental conditions have recently been studied in a the strong contribution of vehicle effect on per-
large database of 536 flux data points from excised meability. Each of the factors for experimental
human skin (Samaras et al. 2012). The QSARs condition was then added to the regression
that were developed account for different dosing equation, whereby skin thickness and exposure
conditions, vehicles, membrane thickness, and type were found to be statistically significant
sample pretreatment. Vehicle properties were variables. With the RT models, inclusion of
computed by weighted averaging. For melting BP - MP ( mix ) had the lowest mean absolute
and boiling points, solute effects of boiling point error (MAE) on the full dataset. These results
elevation and freezing point depression were fac- clearly indicate the importance of accounting
tored in. Models were estimated by stepwise for the vehicle mixture.
regression as well as regression tree. Regression It has also been demonstrated that results from
tree models split the problem space according to one model system can extend to predictions in
covariate regions (leaves) and fit simpler models another model system. The MCF array has been
(often just the mean of the data) in each leaf. The used to predict absorption in IPPSF in a study
RT models were used to investigate the effects of that tested the effect of two surfactants (SLS and
experimental conditions, but only one condition at linear alkylbenzene sulfate (LAS)) on penetra-
a time. That is, no model included all of the exper- tion of six compounds (Riviere et al. 2010).
imental condition effects, possibly because the Penetrant partition coefficient from each vehicle
response space would become too fractured, into MCFs coated with PA, PDMS, and CW was
resulting in subsets too small to estimate parame- determined by mass balance. The partition coef-
ters for even simple models. ficients were then used in a QSPR (6.6) to predict
Neither stepwise regression nor regression log AUC ( R 2 = 0.718 ) .
tree models selected variables for skin thick-
log AUC = I + a ( log K PDMS ) + b ( log K PA )
ness, infinite vs. finite dosing, prehydration of
the membrane, or occlusion. The failure to auto- +c ( log K CW )
(6.6)
matically select for what were found to be oth-
erwise statistically significant effects may be The rank ordering of absorption, penetration, and
due to the large number of descriptors (375) peak flux across the three mixtures in IPPSF was
used in the automated procedure. The most sig- consistent: water > SLS > LAS, except for sima-
nificant variable selected in both models was zine, where SLS > water > LAS. Remarkably, rank
donor concentration, with other descriptors for ordering of log KMCF followed the same pattern
molecular size, hydrophilicity, lipophilicity, and (water > SLS > LAS) except for simazine in all of
polarizability. The regression equation also the MCFs, and propazine in just the CW MCF,
selected the difference between vehicle boiling where the ordering was water > LAS > SLS. Seeing

6  The Effects of Vehicle Mixtures on Transdermal Absorption 113

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...5 Effects of Branched Alkyl Chain.............3.......... USA approach has been employed in these studies (Kim © Springer-Verlag Berlin Heidelberg 2017 119 N......2............ enhancers have been studied (e.... Higuchi Our laboratory has been studying the Pharmaceutics and Pharmaceutical Chemistry... Mechanistic Studies of Permeation Enhancers 7 S... 135 7.3.... more than a decade...... 128 7.. 129 Most of the chemical permeation enhancer stud- 7................ 120 7.............. lar structures and their effects as permeation 3225 Eden Ave....... Li (*) ferent relationships among the enhancer molecu- Division of Pharmaceutical Sciences.......... 127 7...... University of Utah................ Under this approach.........3................ Smith and Maibach 1995..... Rm 160........ 120 System in Transdermal Drug Delivery.... USA Lee et al.. reviewed in OH 45267-­0004.)............. 133 7.... 126 7...................1007/978-3-662-53270-6_7 ..............g.........7 Transport Rate-Limiting Domain nature of the enhancers and their effectiveness in and Equilibrium Partitioning Domain.. 124 References................... College of Pharmacy. W.....................3.....2 Transport Experiments.edu Hadgraft 2001).....3.. 1991.....2 Methods.. 126 7....I........................4 Effects of Hydrocarbon Chain Carbon–Carbon Double Bond........... 119 Molecular Sizes................6 Equilibrium Partition Enhancement ies in the past decades have been aimed at gaining of ES into SC Intercellular Lipids.....K.... Dragicevic..I.2 Effects of Alkyl Chain Length...1 Introduction.............. Maibach (eds....3.li@uc... A different experimental UT 84112........ 130 better insights into the relationship between the 7....3... and the effectiveness of the enhancer compared to a control is determined by the rate of transport of the drug............. Salt Lake City..............1 Isoenhancement Concentrations and Enhancer Effects..........2............... In typical in vitro stud- ies of chemical permeation enhancers. Skaggs Hall.............1 Animal Model........... University of Cincinnati............8 Effects of Permeation Enhancement on Permeants of Different 7.......3 Results and Discussion.....9 Permeation Enhancers in a Nonaqueous 7..............2... ­mechanism of action of permeation enhancers for College of Pharmacy...3 Partition Experiments....3... 126 7.... 131 permeation enhancement.... Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin.............3 Effects of Polar Head Functional Groups. H.................. the enhancer in question is usually applied with a drug in solution or suspension to one side of the membrane............ 121 Conclusion.... Cincinnati....... the dif- S................ 30 S 2000 E.. e-mail: kevin. DOI 10... 134 7. Kevin Li and William I..................3........................... Higuchi Contents 7....1 Introduction 7...... 131 7........

1992. HMS has relatively constant lipid content enhancer in enhancing transdermal transport (Yoneto et al. well absorbed. to determine the intrinsic He et al. HMS was selected as coefficients obtained for the permeants can be the model for human skin for the following rea- used directly to determine the effectiveness of an sons.120 S.K. the permeability and other visceral debris. . and to establish a collected directly. oidal pathway and for the mechanistic studies of meant activity alteration upon transport in the the effects of skin permeation enhancers upon this presence of the enhancers are taken into account. In the symmetric configuration. and a large body of HMS (enhancer potency). 1998). The results suggest that for solution with respect to that in the buffer solution the assessment of skin permeation across the lip- (the control) are corrected for. HMS can be a reliable model for human Taking the above issues into consideration. pore transport pathways using model permeants of 1997). 1989). in the investigation of chemical ation enhancers and for establishing a permeation enhancers for the lipoidal pathway structure–enhancement relationship. Li et al. they cannot be systematically investigated similar to that of human skin (Lampe et al. diffusion cell and in equilibrium with the mem- brane. 1992. However. is not a good model of human skin (Lambert et al. HMS and the membrane is therefore necessary in obtain. direct comparisons of our results with those in Because they are water-insoluble. For example. 7. 1991. if mechanistic insight is to be potencies of the enhancers. This chapter is a review of our studies enhancer is present at equal concentrations in both employing this strategy and the experimental the donor and receiver chambers of a side-­by-­side approaches. 1992. the complications arising from enhancer concentration (or activity) 7. Experiments were conducted with freshly sepa- ment varies with the position within the membrane rated hairless mouse skin (HMS) obtained from and make mechanistic data analysis difficult. First. crepancies between the mechanisms of action of nistic insights into the effects of enhancers upon permeation enhancers in HMS and human skin. 2003. conveniently for structure–enhancement activity. The quantitative structure–enhancement ation enhancement. in the assessment of skin perme. Under these conditions. lipid composition (Grubauer et al. Warner et al. Chantasart and Li 2010) can be avoided. 1989) is also tion. Yoneto et al. 1995. t­ransport across skin. There is no direct evidence of significant dis- have developed a research strategy to gain mecha. such as for experiments requiring Establishing equilibrium between the enhancer long-term skin stability in aqueous solution. previous studies.I. HMS can be an adequate quantitative enhancement on transport across the lipoidal and model for human skin (Kim et al. good enhancers are data is available in the literature allowing the usually lipophilic and relatively water-insoluble. Higuchi et al. tion. Second. so the effects of per. ing mechanistic insights into the action of perme. changes in the chemical relationships of the enhancers in HMS and HEM potential (activity) of the permeant in the enhancer were also compared. 1983). HMS stratum corneum (SC) and need to be solubilized for effective presenta. pathway. Li and W. model and where relatively short experimental times are analyses to separate the effects of permeation involved. a symmetric and equilibrium quantitative structure–enhancement relationship configuration (with respect to the enhancer) should between the enhancers and permeation enhance- be used. 2004). the ment. we skin. With the abdomen region and freed from adhering fat the symmetric configuration.1 Animal Model These enhancer gradients would otherwise lead to a situation in which the local permeation enhance. Chantasart et al. Third. In certain cases.2. Last.2 Methods gradients across the membrane (Liu et al. The effects of pared the effects of permeation enhancers on the the enhancers upon the intercellular lipoidal and permeability coefficients of the lipoidal pathways pore pathway transport have been delineated in of HMS and human epidermal membrane (HEM) order to understand the mechanism of action of the under the symmetric and equilibrium configura- enhancers. (2007) com- different polarity have been used.

A dis- was less than 5 % in most cases and less than advantage of this approach is the inability to con- 15 % in the extreme case at the end of the trans. skin with liquid enhancers and the subsequent the enhancer solution in both diffusion cell cham. To philic enhancers. Scotch tape prior to excision and assembly into The skin membrane was sandwiched between the the diffusion cell.2. Shaker chemical enhancers and evaluate enhancer effec- et al. Warner bination (PD/E) was obtained in the same manner. two half cells and an enhancer solution in PBS Another method to evaluate the potency of was pipetted into both chambers. 2003. In other meant.method was recently used to study the structure– cantly interacts with the SC nor acts as a perme. With highly 1-dodecyl-2-pyrrolidone).2 Transport Experiments permeant tetraethylammonium ion (TEA) were conducted in essentially the same manner. an aqueous tration can be maintained in the SC over the reservoir system (enhancer solubilizing system duration of the permeability experiments.g. b).e. Each compartment has a 2-mL vol. always at saturation in the Corticosterone (CS) was the main model per. ness HMS as described for a typical permeation Permeability experiments with model ionic polar experiment under the symmetric and equilibrium . 7. its solubility was added to the donor chamber fol- lowing enhancer equilibration..2 Reversibility Study withdrawn from the donor and receiver chambers Diffusion cells were assembled with full thick- at predetermined time intervals and analyzed. the depletion of the enhanc- depletion of the enhancers in the aqueous phase ers from the skin to the aqueous solution in the that made it difficult to achieving an equilibrium diffusion cell chambers is expected to be mini- of the enhancers between the aqueous phase and mal. and essentially constant enhancer concen- the SC at the target concentrations. this approach cannot be used to evaluate were the two other steroidal permeants tested in the effects of enhancer concentration upon skin the studies..67 cm2.1 Permeability Coefficient total permeability coefficients (PT) were deter- Determination mined from data obtained under steady-state con- The permeability experiments were carried out as ditions (experiment time points around three to previously described with a two-chamber side-­ five times longer than the lag times).aqueous solution) and hence in the skin. The loss or depletion of all the enhancers solubility of the enhancer in the SC lipids).tiveness under the condition when the SC was tions of the enhancers in the diffusion cell saturated with the enhancer at thermodynamic ­chambers were frequently checked by HPLC or activity equivalent to its pure state (i.7  Mechanistic Studies of Permeation Enhancers 121 7. Warner et al. at the GC. et al. is the direct equilibration of the attain equilibrium of the enhancer with the HMS. Chantasart and Li 2012) will not be dis- Molecular Sizes.skin transport experiments with PBS as the donor bers was replaced until the SC was essentially in and receiver solution in a side-by-side diffusion equilibrium with the enhancer solution (enhancer/ cell in the absence of cosolvents or solubilizing PBS).permeation enhancement. 2003).e. due to the extensive lipophilic enhancers. A list of the permeation enhancers.2. Permeation enhancer cussed in the section on “Effects of Permeation studies using this approach (Ibrahim and Li Enhancement on Permeants of Different 2009a.2. Samples were 7.2.trol the enhancer concentration in the diffusion port experiments.enhancement relationship of highly lipophilic ation enhancer was used (Warner 2003. Other permeants used will be dis. agents (Ibrahim and Li 2009a.1. This of micelles or cyclodextrin) that neither signifi.but the skin was stripped 30 times with 3 M ume and an effective diffusional area of 0. The perme- by-­side diffusion cell in phosphate buffered ability coefficient of the dermis–epidermis com- saline (PBS) at 37 °C (Yoneto et al. cell chamber (i. The concentra. 1995.2. 2008). With highly hydrophobic enhancers (e. particularly highly lipo- enhancers investigated is shown in Fig..” The steroidal permeant at cussed in this chapter but have been discussed in radiotracer levels and at concentrations far below a recent book chapter (Li and Higuchi 2015). b. Estradiol (ES) and hydrocortisone (HC) words. The 7.

I.2-alkanediols (AD) HO O O HO N HO HO R O R n-alkyl-b-D-glucopyranosides (AG) N.3-dioxolanes (MD) 1-alkanols (AL) O OH N R HO R 1-alkyl-2-azacycloheptanones (AZ) 1.N-dimethyl alkanamides (AM) OH O OH R R OH O 1. R′ = alkyl chain N R N R 1-alkyl-2-pyrrolidones (AP) 1-alkyl-2-piperidinones (API) R R HO O O 2-(1-alkyl)-2-methyl-1. Higuchi Fig.1  Chemical structures of O O the enhancers.122 S. Li and W.K.2-dihydroxypropyl alkanoates (MG) trans-3-alken-1-ols (TAL) R OH R OH R' cis-3-alken-1-ols (CAL) Branched alkanols (bAL) . R. 7.

the in PBS after enhancer pretreatment were within a use of CS as the probe permeant allows Eq. PBS and in PBS.1) and TEA.2. (7. and their effects PL + PP PD / E upon permeation across SC were shown to be essentially reversible (permeability coefficients Based on the results from previous studies.3 Model Description and Analysis For other steroidal permeants.3) bers of the diffusion cell were then rinsed with PBS to remove the enhancer equilibrated in the where PL and PP are the permeability coefficients membrane. for the lipoidal pathway and the pore pathway transport studies were carried out with PBS in (TEA is used as the probe permeant for estimat- both chambers.3) into Eq.1) E= (7. 2003. time plot. and dQ/dt is the slope of the linear region of is enhancer/PBS and PBS. in the obtained with PBS after pretreatment with SC.X and PL. 2001. CD is the concentration in the donor cham. both cham- PSC = PL + PP (7. 2001): dal pathway transport enhancement factor (E) is: 1 dQ PL . Chantasart et al.4) 1 1 E = 10 (Warner et al. respectively. respectively.2. (7. O SO where A is the diffusional area of the diffusion where PL. 1992). Determination ity coefficient for the epidermis-dermis combina.4) factor of 2 of those in PBS without enhancer to be approximated by: pretreatment). He et al.6) ACD dt PL .7  Mechanistic Studies of Permeation Enhancers 123 configuration. X S X P= (7. and equilibrium between the mem. respectively. PT » PL (7. cients for the lipoidal pathway when the solvent ber. 2004). The intercellular lipid domain in SC is gener- enhancers were then compared with those ally accepted as the lipoidal transport pathway obtained with pretreatment with PBS only.2) Except for the highly lipophilic enhancers.4) with PD/E and PP values obtained The permeability coefficient (P) of a probe per. into parallel lipoidal and pore pathway compo- brane and the enhancer solution was allowed to nents in SC via the following equation: take place. and SX the cumulative amount of permeant in receiver and SO are the permeant solubilities in enhancer/ chamber (Q) vs. where PSC is the permeability coefficient for the 7. However.2. PSC can be further divided adding excess crystals of the permeant into the .5) 7. respectively. (7. PL can be calcu- of Experimental Data lated by Eq. (7. across SC. Following the PBS rinsing regime. (7.2. in this protocol. The solubilities of the steroidal permeants in PBS tion (D/E) and can be obtained from experiments and the enhancer solutions were determined by of tape-stripped skin. from transport experiments with stripped skin meant was calculated according to Eq. The solubility ratio Total permeability coefficient expression for corrects for any activity coefficient differences full-thickness skin is written as follows: between the activity coefficient in PBS and that in the enhancer solution. Use of the solubility 1 PT = (7. + 2004. The equation for the lipoi- (Warner et al. 2003.O are the permeability coeffi- cell.2) ratio assumes that Henry’s law is obeyed for the 1 1 + permeant in both PBS and enhancer solutions PSC PD / E (Kim et al. all yields: enhancers were tested for reversibility at enhancer 1 concentrations up to enhancement factor (E) at PT = (7. Substituting Eq. The permeability coefficients ing the magnitude of PP).4 Permeant Solubility stratum corneum (SC) and PD/E is the permeabil.

concentration) containing trace amounts of radio- pared according to the method described by labeled ES (3H-ES) in a screw-capped glass vial. the drug suspension was shaken for 72 h at 37 °C. 7. This rinsing protocol (the number of rinses fusion cell (Yoneto et al. Briefly.2. Heptane-treated SC (about 1–2 mg) or delipidized ments with nonpolar organic solvent were also SC sample was carefully weighed and equili- shown to remove skin surface lipids (e. the latter to test whether residue of SC was then rinsed several times with Henry’s law is obeyed in the two liquid phases. 1993.2. Li and W.3. from both phases and appropriately diluted for sub- sequent analysis using HPLC or GC.3.5 Determination of Partition pidized HMS SC was prepared according to the Coefficient in Bulk Organic method described previously (Yoneto et al.2 % trypsin in nary studies showed that equilibrium of enhancer PBS solution up to the surface of SC. Both the organic and aqueous phases were due was carefully weighed and used for the parti- centrifuged. Nicolaides 1974). culture tubes were then centrifuged for 15 min at 3500 rpm. The E = 10 concentration. dermis enhancer solution evaporation and put in a water side down.124 S. 7. the dermis the partitioning experiments. dried n-heptane-treated SC sam- Organic solvent/PBS partition coefficients were ples (about 1–2 mg) were weighed and trans- obtained at the aqueous enhancer concentrations ferred into 5 mL CHCl3/MeOH (2:1) mixture and corresponding to E = 10 and at one tenth of the equilibrated for 48 h at room temperature. After drying. and the clear supernatants were ana. 1998) and would not be and the rinse time) was shown to remove approx.I.1 n-Heptane Treatment and SC ation enhancer and of probe permeant ES into preparation n-heptane-treated or delipidized HMS SC.g.3. The old setup used a Franz dif- ­lipids. The following is a brief descrip- imately 20 % of the SC lipids but did not disrupt tion of the other method (Chantasart et al. HMS was rinsed with different partition experimental setups were used heptane for 3 × 10 s to remove the SC surface in our laboratory. Heptane-treated HMS SC samples were prepared as described in the previous section.3 Partition Experiments Partition experiments were carried out to deter- mine the uptake amounts of the chemical perme- 7. When the skin membrane was placed in dis. The water. discussed here. Abrams brated in about 20 mL of enhancer solution (E = 10 et al. The fresh CHCl3/MeOH (2:1) mixture and dried two-phase systems were maintained at 37 °C for under room temperature for 24 h. Solvent/PBS Systems 1998). The SC was kept in a freezer for later use.2. Similar treat. The with distilled water several times and swabbed enhancer and ES concentrations of the solution in with Kimwipe® tissue paper to remove excess the screw-capped glass vial were checked. on a filter paper (quantitative filter bath with shaking at 37 ± 0..2 HMS SC Delipidization lyzed for permeant concentrations with HPLC. Higuchi enhancer solution in Pyrex culture tubes. SC was then pre. The and dried at room temperature. the SC was placed on aluminum foil wet SC sample was carefully weighed in a .2. Two Before SC preparation.K. The dried resi- 72 h. 2004). The Petri and 3H-ES with the SC sample took place in less dish was covered and maintained at 37 °C for than 12 h and that a longer incubation period 16 h. Whatman®) mounted on a Petri dish. The SC was then rinsed zers and blotted by Kimwipe® tissue paper. The deli- 7.1 °C for 12 h. and aliquots were carefully withdrawn tion experiments. the SC and viable epidermal layers would separate and sample was taken out from the solution by twee- fall away from the SC. (1998).2. the SC barrier (He et al. Kligman and Christophers (1963) and Yoneto The vial was sealed with parafilm to prevent et al. After 12 h. Briefly. might result in too fragile a membrane sample for tilled water after the trypsin treatment.2. 2003).3 Partition Experiments with Heptane-Treated and Delipidized HMS SC 7. incubation period was chosen because prelimi- The Petri dish was filled with 0. Then. the skin was placed. 1. The 12-h paper No.

This assumption was consid- and the concentration of the enhancer in aqueous ered to be reasonable because previous studies bulk phase (Ci). temperature with occasional gentle agitation.9) Corneum Intercellular Lipids An important question raised in the above trans. diffusion. Furthermore.4 Permeant Partitioning were analyzed using the model to obtain the par- into the Transport Rate-­ tition coefficient (KSC) and diffusion coefficient Limiting Domain (DSC) of SC. However.3. The reduced parameters KSC′ and and Equilibrium Permeant DSC′ of SC were then calculated: Partitioning into the Stratum DSC¢ = DSC / L2 (7. nonsteady state ES transport analysis was complicated. chamber concentration was kept at sink condi- tions. The solubility ratio corrects for any activ. ES and CS for permeation across the lipoidal culated as follows: pathway of HMS SC (Yoneto et al. A correction for the enhancer in the equilibrium partitioning enhancement of ES. for ES in the following study. The strategy here Wdry is the dried heptane-treated or delipidized was to examine the relationship between the SC weight. The skin transport model (He et al.Wdry ) (7. 1995). which was true in all transport experi- coefficient of ES in PBS and that in the enhancer ments carried out in the study. The the partition enhancement in SC permeation was extracted solution was then transferred to a screw. assumes that both SC and D/E are homogenous C′i is the concentration of 3H-ES in aqueous and divides the SC and D/E into a sufficient num- bulk phase. K SC ¢ = K SC L (7.7) was observed in ES transdermal permeation. i = . ES in enhancer solution and in PBS.Wdry ) Ci¢ ùû / Wdry S ¢ two-layer numerical transport simulation with a K ES = X (7. Salt Lake City. and the subscript i represents the transport partitioning enhancement of CS and the enhancer. 5 mL of 100 % equilibrium partition enhancement data of ES a ethanol was added into the bottle to extract the direct correlate of the partition enhancement of enhancer and ES from the sample for 48 h at room ES in SC permeation? To address this question.(Wwet . experiments and those data obtained in equilib- The uptake amount of enhancer in the heptane-­ rium partitioning experiments of ES was not treated SC or delipidized SC was calculated as practical due to the D/E layer being a significant follows: barrier for ES permeation across HMS.2. significant ES metabolism Aextracted . Wdry Wdry Because of these difficulties. with the assumption that ES and CS should likely lated according to the wet weight of SC (Wwet) behave similarly. and dimension parameters. i Ci Acorrected . fuged at 3500 rpm for 15 min. respec. a direct comparison of the parti- was analyzed for the enhancer by GC or HPLC tion enhancement data obtained in transport and for ES by a scintillation counter.8) C¢ i SO¢ least squares-fitting software Scientist (MicroMath. 2005) is a é Aextracted ë ¢ . The test tube was centri. and it was where Aextracted. determined in skin transport experiments using a capped Pyrex test tube. aqueous compartment(s) of the SC was calcu. Then. The partition coefficient of ES had shown similar permeability coefficient (KES) for partitioning from the aqueous phase into enhancement effects of chemical enhancers with n-heptane-treated SC or delipidized SC was cal.7  Mechanistic Studies of Permeation Enhancers 125 s­nap-capped glass bottle.10) port and equilibrium partition studies was: is the .(Wwet . i is the amount enhancer extracted decided to employ CS as the surrogate permeant from heptane-treated or delipidized HMS SC. The permeant con- tively. centration in the donor chamber was assumed ity coefficient differences between the activity constant. The receiver solution. nonsteady state transport analysis (He et al. The transport data of full-thickness HMS 7. This model where A′extracted is the amount of extracted 3H-ES. The supernatant 2005). and S′X and S′O are the solubilities of ber of layers characterized by partition. UT).

1 1 10 Carbon number Enhancer concentration (%w/w) Fig. and deviation because the error bar generally lies within the 1-octyl-2-pyrrolidone (OP).3 (Warner et al. The lipoidal pathway with 1-butyl-2-pyrrolidone.3. Each data point rep- SC lipoidal pathway in the presence of 1-butyl-2-­ resents the average value without showing the standard pyrrolidone (BP). aqueous enhancer tional group but different alkyl chain length) in the concentration plots were observed for the enhanc.2 Effects of Alkyl Chain Length obtained with the enhancers at E = 10 by those with PBS control.I. and HC permeation across the SC constant permeant thermodynamic activity).55 translates into an around AP AL API AZ AD ES BP CS BP HC BP 1 AM AG MD MG ES HP CS HP HC HP 15 Aqueous concentration (M) ES OP CS OP HC OP 0.00001 0 2 4 6 8 10 12 14 0. 7. essentially the same for the steroidal permeants culty and uncertainty in assigning the L value and of different lipophilicity. and 1-octyl-2-­pyrrolidone is a slope of around −0.6) in Fig.K. 7. figure.001 5 0. and hydrocortisone (HC) across the number of the enhancer alkyl chain.3. The enhancement of KSC′ and DSC′ (EK.1 . (7. These reduced parameters KSC′ and DSC′ were increasing aqueous enhancer concentrations are defined (Okamoto et al. The enhancement factor profiles at SC.1 Isoenhancement tor and in this case E = 10. series (enhancers have the same polar head func- Similar enhancement factor vs. 1995). The transport enhancement symbol in the plot. steroidal permeants. Li and W. 7. Higuchi where L is the effective path length across ers studied. 2003).55 found for each enhancer as permeation enhancers (Yoneto et al. isoenhancement con- 7.2  Transport enhancement factors of estradiol (ES).01 0.2 shows a representative plot of enhance. The value of −0. 7. 1-hexyl-2-pyrrolidone (HP).2.3  Relationships between the aqueous E = 10 isoen- Fig.0001 0. 126 S. Figure 7. Enhancer abbreviations are provided factors were calculated using Eq. 7. respectively) was cal- culated by dividing the KSC′ and DSC′ parameters 7. These isoenhancement Concentrations and Enhancer concentrations were interpolated from the E vs.1 Enhancement factor. between the E = 10 enhancer concentration and the ment factor vs. major conclusion deduced from the data in Fig. CS. 1988) to avoid the diffi. SC and ED. hancement concentrations of the enhancers and the carbon corticosterone (CS). aqueous enhancer concentration carbon number of the enhancer n-alkyl group (at for ES.01 10 0.3 1-hexyl-2-pyrrolidone. The isoenhancement concentrations at E = 10 for more than 20 different enhancers are presented in Fig. 7. E 0.3 Results and Discussion centration is defined as the aqueous concentrations of enhancers to induce the same enhancement fac- 7. Effects aqueous enhancer concentration plots similar to those in Fig. SC. mental transport data. suggesting the same to minimize the number of parameters for least mechanism of permeation enhancement for these square fitting in model analyses of the experi.3 shows the relationship Figure 7.

g. Smith and Maibach 1995). Enhancer abbreviations are pro.g. tions in the intercellular lipid domain of the SC membrane 2003). tenfold lower than those of 1-alkyl-2-­pyrrolidones The results of the equilibrium partition experi. 7. azacycloheptanones are consistently around Tanford 1980). 7.3. 7.4.4  Relationship between the enhancer concentra.g. potency based on the concentrations of the tions in Fig. tional group of an enhancer upon its effectiveness in transdermal permeation enhancement (e. enhancers as compared with the pyrrolidone tions (He et al.4 is the their concentrations in the aqueous phase in the same as that in Fig. group of the 1-alkyl-2-pyrrolidones based on Note that the scale of the y-axis in Fig... 7. The polar functional groups are more effective (more constant slope of −0. 7.1 groups of the enhancers act only to transfer the enhancers from the aqueous phase to the hydro- 0. In particular.5-fold when the alkyl chain length of the The data in Fig. esting finding because studies using conventional generally believed to be the SC intercellular lip.3 E  ffects of Polar Head for the enhancers. In other words. 2003.5-fold increase in potency per methylene group 7. it has been suggested that the azacycloheptanone func- AP AL API AZ 100 tional group is more potent than other polar head AD AM AG functional groups in general due to specific inter- 10 actions between the functional group and the (micromole/mg membrane) Membrane concentration ceramide lipid matrix (e.3). 7.  7.4 based on the concentrations of the enhancers in reveals that there is little effect of the enhancer the intercellular lipid lamellae (relative to that polar head functional group upon the enhancer based on the E = 10 aqueous enhancer concentra. 7. partition coefficients of the enhancers with a vided in Fig. 7.3 to demonstrate a structure-enhancement Fig.7  Mechanistic Studies of Permeation Enhancers 127 3. however. at the same n-alkyl chain length.4 sug. suggesting that ments with the enhancers conducted to determine the azacycloheptanone group makes the 1-alkyl-­ the amount of enhancers in the SC intercellular 2-azacycloheptanones more effective permeation lipids under the isoenhancement E = 10 condi. Hadgraft et al.3 also show that some enhancer enhancer decreases by one methylene group.3.1 slope of around −1 suggests that the potencies of . The data in Fig. donor and receiver chambers. Each data point rep- hancement concentrations and the octanol/PBS resents the average value. The data in Fig. the rela- gest that there was little effect of the enhancer tive constant concentration of enhancer uptake alkyl chain length upon the enhancer potency into the SC lipid domain at E = 10  in Fig.4. enhancers at their site of action. 1996). 7. For example. This is an inter- sic potency of the enhancers at their site of action. the aqueous Functional Groups concentration required to induce E = 10 increases 3. imply that the polar head and alkyl 0. However.5 is a replot of the data shown in Carbon number Fig. The correlation between the E = 10 isoen- at the E = 10 isoenhancement conditions and the carbon number of the enhancer alkyl chain. 7. relationship for the enhancers (Warner et al.. is relatively independent of their alkyl group onstrated the influence of the polar head func- chain length and lipophilicity. 2004) are shown in Fig. 2 4 6 8 10 12 14 Figure  7.01 carbon phase of the SC lipid lamellae and make available the enhancers for their action in the SC 0. thus suggesting that the intrin. the E = 10 isoen- transfer of the methylene group from the aqueous hancement concentrations of 1-alkyl-2-­ phase to a relatively nonpolar organic phase (e. experimental methods in the literature have dem- ids.001 transport rate-limiting domain. Brain and Walters 1 1993.55 for the different enhancer potent) than the others in inducing permeation series suggests a hydrophobic effect involving the enhancement.

128 S. In the equilibrium partition experiments of the centrations of the cis-and trans-3-alken-1-ols cis. Each data point represents the average value with.3. resents the average value without showing the standard out showing the standard deviation because the error bar deviation because the error bar generally lies within the generally lies within the symbol in the plot. Enhancer symbol in the plot.I.5 5.4 E  ffects of Hydrocarbon Chain the enhancers is taken into consideration.  7. 7.3 are also included related to the enhancer lipophilicities. when limiting domain has a microenvironment with the E = 10 isoenhancement concentrations of the polarity similar to the polarity of bulk octanol.and trans-3-alken-1-ols and of the n-alkyl These hypotheses and the results of Fig. 7. 7.6 for comparison. 7. cient here is consistent with the demonstrated bon bond on the n-alkyl chain of an enhancer structure–enhancement relationship for the with a carbon–carbon double bond have been n-alkyl enhancers in Fig. 7. and the E = 10 isoenhancement con.5 2. Together in Fig. Enhancer abbreviations are provided abbreviations are provided in Fig. it is seen with the data analysis in Figs. the site of action through a free energy of transfer the results in Fig.5.5  Correlation between the aqueous E = 10 isoen. 7. Fig.5 4.6  Relationships between the aqueous E = 10 isoen- hancement concentration of the enhancer and its octanol/ hancement concentrations of the enhancers and the carbon PBS partition coefficient (Koctanol/PBS).0001 0. 7. the (Li and Higuchi 2015).6 would suggest the cis.5) when the lipophilicity of 7.001 −4 −5 0. (b) the polar head that the E = 10 isoenhancement (aqueous) con- group assists the translocation of the enhancer to centration is a measure of the enhancer potency. 7. 2004).and trans-3-alken-1-ols.7. Based on the criterion rate-limiting domain in SC. The Carbon–Carbon Double Bond correlation between the E = 10 isoenhancement concentration and octanol/PBS partition coeffi- The effects of substituting a single carbon–car.6 (He et al.and trans-3-alken-1-ol data fall closely on the correlation line in the figure (same correlation line as that in Fig. The data for the AL 1 CAL Log (aqueous concentration) (M) 0 AP AL API AZ AD TAL Aqueous concentration (M) AM AG MD MG 0. investigated. cis.01 −3 0. In Fig.and from the bulk aqueous phase to the transport rate-­ trans-3-alken-1-ols are less potent than the limiting domain. 7. The slope of the line number of the enhancer alkyl chain. Each data point rep- is −1.5 2 4 6 8 10 12 Log Koctanol/PBS Carbon number Fig.6.1 in Fig.5 have enhancers are plotted against their octanol/PBS been discussed in detail in a recent book chapter partition coefficients (Koctanol/PBS) in Fig.5 1.K. Li and W. it is that the E = 10 isoenhancement concentrations of reasonable to hypothesize that (a) permeation the cis. 7. 7. times higher than those of the corresponding meation enhancer to partition into the transport n-alkanols (open squares). 7.5 3. 7.3 and 7. cis. However.4.1 −1 −2 0.and trans-3-alken-1-ols are two to three enhancement is related to the ability of the per. Higuchi the enhancers for the steroidal permeants are n-alkanols reported in Fig. the concentrations (closed symbols) are plotted against the carbon of the enhancers in the SC intercellular lipid numbers of the enhancer hydrocarbon chains in Fig. and (c) the transport rate-­ n-alkanols by a factor of 2 to 3.1 .

5 2 4 6 8 10 12 14 1 1. based on the concentrations of the enhancers in the SC intercellular lipid domain. 3-alkanols.  7. 7. deviation because the error bar generally lies within the vided in Fig. Aungst 1989. the isoenhancement concentrations at Log Koctanol/PBS E = 10 of the branched alkanols (closed symbols) Fig. −4 2-­ heptanol. and 4-alkanols are 2-hexanol. 7. 1987.1 enhancers in Fig. Enhancer based on the data for more than 20 n-alkyl abbreviations are provided in Fig. The slope of the line is −1.5 2. The slope of the line is −1 . −5 4-octanol. Each data point rep- number of the enhancer alkyl chain. Tenjarla et al. resents the average value without showing the standard resents the average value. are plotted against their Koctanol/PBS values.5 3. 2004).1 −2 −2. 3-octanol. Brain and here has little effect upon the enhancer potency Walters 1993. 3-alkanols.5 Fig. hancement concentration of the enhancer and its octanol/ the data of the n-alkyl enhancers (including the PBS partition coefficient (Koctanol/PBS). 7.9  Correlation between the aqueous E = 10 isoen- tions in the intercellular lipid domain of the SC membrane hancement concentration of the enhancer and its octanol/ at the E = 10 isoenhancement conditions and the carbon PBS partition coefficient (Koctanol/PBS).1 symbol in the plot. 2-nonanol. n-alkanols) are included in Fig.01 −3 0. Enhancer abbreviations are pro. This is some- what surprising because enhancers with unsatu. 7.5 5.9. 7.5. Again.3. In 0.5 3 3.8  Relationship between the enhancer concentra. Log (aqueous concentration) (M) 0 AD AM AG MD 4-alkanols.5 0. and 4-­heptanol. 1999). 3-heptanol. Each data point rep. the AP AL API AZ AD 10 2-alkanols AM AG CAL TAL 3-alkanols (micromole/mg membrane) −1 Membrane concentration Log (aqueous concentration (M)) 4-alkanols 1 5-nonanol −1. 3-hexanol. Fig. 7. and 5-nonanol) were also investigated MG CAL TAL −1 as another group of skin permeation enhancers to provide insights into the mechanism of action of −2 both n-alkyl and branched chain enhancers −3 (Chantasart et al. and 4-nonanol.8). 7  Mechanistic Studies of Permeation Enhancers 129 domain under the isoenhancement E = 10 potent than enhancers with saturated hydrocar- ­conditions are essentially constant (Fig.5 4.001 −3. respectively.7  Correlation between the aqueous E = 10 isoen. 3-nonanol. 7.5 E  ffects of Branched Alkyl rated hydrocarbon chains are expected to be more Chain AP AL API AZ Branched chain alkanols (2-alkanols.5 1.9. The 2-alkanols. Each data point rep. 7. 7.5 2 2.5 n-alkyl enhancers 0. The bon chains based on molecular modeling of skin substitution of a single carbon–carbon bond with permeation and previous experimental results a carbon–carbon double bond on the alkyl chain (Golden et al. Different from the random deviations for the n-alkyl enhancers (crosses). 2-octanol. and the resents the average value without showing the standard deviation because the error bar generally lies within the straight line shown in the figure is a best fit line symbol in the plot.5 4 Carbon number Log Koctanol/PBS Fig.

same and independent of alkyl-chain branching.01 branched chain alkanols from the n-alkyl chain enhancers. the Koctanol/PBS val- tion increases with enhancer lipophilicity. branched chain alkanols.8). Each data point represents the average value the n-alkanols. 2003. 7. These deviations support the view. Li and W.5 4.130 S.1 alkanols suggested with the data in Fig. modest but consistent positive deviations (in the direction of lower potency) from the quantitative structure–enhancement correlation (the straight 7. Despite the observed deviation of the 0.to sevenfold) was induced under the potencies of the n-alkyl enhancers are essentially isoenhancement conditions of E = 10 for all the the same and independent of their alkyl chain enhancers studied. (7. Figure  7.5 2. 7.6 Equilibrium Partition line).9 in a recent book chapter. The goal here was to determine the enhancement philicity of the enhancers when the hydroxyl of the partitioning of a lipophilic permeant into group moves from the terminal end towards the the SC intercellular lipids under the isoenhance- center of the enhancer alkyl chain. the ing suggests that (a) the same target site in the SC . The relatively constant four- length. The KES values were deter- Figure  7.10  Relationship between the enhancer concentra.5 5. Higuchi 2-alkanols c­ oncentrations of the branched alkanols in the SC 3-alkanols 10 4-alkanols intercellular lipid domain (closed symbols) required 5-nonanol to induce the E = 10 conditions are generally higher (micromole/mg membrane) n-alkyl enhancers than those of the n-alkyl enhancers (crosses in the Membrane concentration 1 figure). the figure. that the branched chain alkanols are slightly less potent than the n-alkyl In addition to the equilibrium partition experi- enhancers. ment E = 10 conditions. Whereas the intrinsic of KES (four. 2004). 2004. intrinsic potency and increasing effective hydro.9. chain alkanols. This result is consistent with the relatively lower intrinsic potency of the branched chain 0.5 coefficient from the aqueous phase to the SC Log Koctanol/PBS intercellular lipid phase (log KSC lipid/PBS) and log Koctanol/PBS continues to hold for the branched Fig.I. branched chain alkanols (closed symbols) show Li and Higuchi 2015). experiments were also E = 10 aqueous concentrations of the branched conducted with a model steroidal compound ES chain alkanols could be attributed to decreasing (He et al. branching of the alkyl chain decreases to sevenfold enhancement in permeant partition- the intrinsic potencies of the enhancers.5 3. Nevertheless. The microenvironment of the tions in the intercellular lipid domain of the SC membrane enhancer site of action remains essentially the at the E = 10 isoenhancement conditions and the octanol/ PBS partition coefficient (Koctanol/PBS) of the enhancers. Intercellular Lipids tor of enhancer potency.10 presents the concentrations of the mined with Eq. aqueous phase into the SC intercellular lipid lished for the n-alkyl enhancers that the potency domain (KES) under the E = 10 conditions of more of an enhancer based on its aqueous concentra. than 20 different enhancers vs. Chantasart et al. 5. based Enhancement of ES into SC on the assumption that Koctanol/PBS is a valid predic.3. approximately the same enhancement ment conditions of E = 10. it should be noted that the correlation between the logarithm of the enhancer partition 0. The dotted line represents branched chain alkanols and n-alkyl enhancers in the KES value in PBS control. plots of the partition coefficients of ES from the tinue to support the hypothesis previously estab.5 1.11 shows the the results of the branched chain alkanols con.001 0. and all other studied enhancers fall on the same regres- sion line (Fig. ues of the enhancers.K. The lower potencies based on the ments of the enhancers. As can been seen in the SC intercellular lipids under the isoenhance.

2005).8 E  ffects of Permeation port across the SC rate-limiting domain and the Enhancement on Permeants partition enhancement in the equilibrium parti. n ≥ 3). n ≥ 3). 7.1 (mean ± SD.6 because the error bars generally lie within the symbols in the plot.8 ± 0. of enhancers presented in a recent book chapter tioning experiments is at the same time the trans. of Different Molecular Sizes tion experiments is required. dence that the rate-limiting domain for the trans- tioning and diffusion.9 and ED.3 ± 0.8 and ED. SC intercellular lipids (in the range of 4–7) is quite ment corresponds to around a 4. Two other steroidal permeants HC .7  Mechanistic Studies of Permeation Enhancers 131 600 AP AZ AL AG API the same as the transport rate-limiting domain for AM AD CAL TAL bAL permeation across SC. The consistency between the partitioning ers. This result also supports the conclusions It would be inappropriate to conclude that the on quantitative structure enhancement relationship uptake domain probed in the equilibrium parti. When the total flux enhancement (E) represents the KES value in PBS control. 7. SC.0 ± 1. This suggests that the transport enhancement of CS was mainly driven by partition enhancement in the rate-limiting domain of lipid lamellae is fluidized by the studied enhanc. 1-octyl- 100 2-pyrrolidone (OP).11  Relationship between the partition coefficients domain of HMS is significantly higher than the of ES (KES) for partitioning from the aqueous phase into enhancement of permeant diffusion coefficient in the SC intercellular lipid domain and the enhancer octa- nol/PBS partition coefficients (Koctanol/PBS). in the trans. 7. (Li and Higuchi 2015). and 1-hexyl-2-azacyclohepta- PBS none (HAZ) (He et al.to 7-fold and significant. lipoidal pathway and the intercellular lipid domain probed in the equilibrium partitioning experiments have similar properties regarding the partitioning 7. Comparison of the partition enhancement in permeant trans. SC around 6–8) and the equi- rate-limiting domain and the enhancer site of librium partitioning enhancement of ES with the action. the ES Partition Coefficient 400 cumulative amount of CS transported across HMS 300 vs. SC was 6. respectively. Enhancer abbreviations are provided in Fig. Most of the work so far presented in this chapter ing would suggest that the SC intercellular lipid was based on the data of a single model steroidal domain probed in the partitioning experiments is permeant CS.3. time profiles in CS transport experiments were analyzed with a transport model to obtain the least 200 squares-fitting KSC′ and DSC′ of CS in PBS.to 2.9 (mean ± SD. port of the model permeants through the SC port rate-limiting domain.5.9 ± 2. and (c) the tenfold permeation enhance. (b) the uptake domain probed in these parti.5-fold enhancement in permeant parti. enhancement of transport found with the SC rate-­ tioning studies is at the same time the transport limiting domain (EK. SC was represents the average and its standard deviation (n > 3). EK. This finding provides quantitative evi- 1. SC was 7. 1. EK. Consistent enhancer effects upon transport and equilibrium partition. The least squares 0 1 2 3 4 5 fittings of the CS transport data were satisfactory. For HAZ with E of The standard deviations of Log Koctanol/PBS are not shown 11. Each data point was 12 for OP. Enhancer Log Koctanol/PBS and the results show that the enhancement of per- meant partitioning into the transport rate-limiting Fig. 500 As described in the Experimental section.3. port rate-limiting domain and the enhancer site of action with only the KES data above.7 Transport Rate-Limiting enhancement effects of chemical permeation Domain and Equilibrium enhancers upon the lipophilic model permeants Partitioning Domain and therefore that these domains are likely to be the same. The dotted line the domain. SC was 1.

and fluidize the SC lipids.Ipomeanol HO O OH HO O OH Kaempferol OH O OH HO OH H H H H H H O HO Hydrocortisone Estradiol and ES were also employed to examine the gen. properties of a permeant can influence the trans.12 Chemical OH OH OH structures of the probe permeants Ethanol Propanol Butanol HO H2N O OH Phenol Benzyl Alcohol Benzamide O O NH OH O HO 2 . Stein 1986. Fig. Higuchi Fig. Xiang and Anderson 1994)..2). To examine the effects of permeant molecular port of the permeant across SC. For example. it is size upon transport enhancement.K. and when enhancers erality of the transport enhancement results. 7. 7. the physiochemical ent molecular sizes (Mitragotri 2001). the increase in the bilayer essentially the same permeation enhancement free volume can have different consequences on was observed with all three steroidal permeants transport enhancement of permeants with differ- (e.. However. different molecular sizes and lipophilicities ation across lipid bilayers (e.g.132 S.Acetamidophenol 4 . Li and W. transport general knowledge that there may be a steep-­ experiments were conducted using permeants of permeant molecular size dependence in perme.g. 7. (Fig.12) under the E = 10 enhancer conditions .I.

Each data point represents the “phase” and itself behaves as an enhancer.4 and 7.3. due to its relatively low .6) and with the assumption that the Higuchi 2015). Further investigation on this Enhancement Factor. This strong enhancer situation with an enhancer concentra- molecular weight dependence is consistent with tion gradient in the SC.13  Relationship between the transport enhance. tively constant concentration across the SC com- Given the results in Fig. 7. seen in the figure. In trans- enhancement factors for ES. E 10 OP subject is required.13. experiments are expected not to be limited only ment factors and the molecular weight of the permeants to the aqueous system. be lowered or raised. Fig. which favors the coefficients PSC and PD/E of the enhancer. this resulting in an asymmetric dence upon permeation enhancement. CS.13. 7. caution needs to be pared with that of the other enhancer (Enhancer exercised in generalizing the results presented in B). shows the SC concentration gradients of two licity upon permeation enhancement was mini. Figure 7. As can be permeation enhancement and permeant lipophi. the absorption of the more licity was observed among the studied permeants lipophilic enhancer (Enhancer A) is largely (provided that the SC lipoidal pathway is the dermis-­controlled and therefore exhibits a rela- dominant transport pathway for permeation). the molecular weight of of the enhancer at its site of action may therefore the permeants under the isoenhancement condi. enhancer permeation essentially the same in Fig. can be found in a recent book chapter (Li and (7.9 Permeation Enhancers 4 in a Nonaqueous System 2 in Transdermal Drug Delivery 0 Although the studies presented in this chapter did 10 100 1000 Permeant molecular weight not include nonaqueous vehicles or conventional cosolvents. the in our study of permeation enhancers. and HC are dermal drug delivery. Further discussion enhancement factors are calculated using Eqs.13 presents the results the partitioning tendency of the enhancer from of the enhancement factors of transport across the the patch vehicle into the SC. The dicted from thermodynamics. For transport enhancement of permeants of large illustrative purposes. and no significant dependency between meability coefficients across the SC. the non- aqueous vehicle or cosolvent may only alter the for CS (Warner 2003). a non- average and its standard deviation (n > 3) aqueous system should not affect the intrinsic potency of the enhancer.14 qualitatively molecular sizes. unless the vehicle is under the E = 10 condition for the steroidal permeants with 1-hexyl-2-pyrrolidone (HP) or 1-octyl-2-pyrrolidone able to partition into the SC intercellular lipid (OP) as the enhancers. However. there occurs across the SC from the transdermal patch is a strong permeant molecular weight depen. 7  Mechanistic Studies of Permeation Enhancers 133 12 this chapter to permeants of different physico- HP chemical properties. the conclusions derived from these Fig. dosage form such as a transdermal patch and alter ers in this study. First. enhancers with different lipophilicities and per- mal. but this effect can be pre- tions: E  = 10 for steroidal permeants. For Enhancer B. As discussed earlier. to the blood sink. In this scenario. 7. presence of the enhancers did not affect the ther. 1-Hexyl-2-pyrrolidone thermodynamic activity of the enhancer in the (HP) and OP were the model permeation enhanc. Another important issue is the symmetric situ- modynamic activities of the permeants in the ation with the enhancers in equilibrium with skin aqueous solution. 7. The concentration SC lipoidal pathway vs. This enhancer concentra- an enhancer-induced increase in the free volume tion gradient is related to the permeability of the SC intercellular lipids. 8 6 7. The effect of permeant lipophi.

bond on the hydrocarbon chain of the enhancer pared. the substitution of a carbon–carbon single metric and asymmetric configurations were com. 7. the effects of enhancers upon the relatively independent of its lipophilicity.2): dotted line. simply applying ers for the lipoidal pathway of the SC have the most lipophilic enhancer does not guarantee been obtained. is related to the enhancer lipophilicity. and PSC for Enhancer A > Enhancer ment will affect the enhancer concentration in SC and B. branching of the suggests that the mechanisms of the enhancers n-alkyl chain of the enhancer generally . (b) the intrinsic asymmetric situation in practice has been exam. The present study supports the success. which is well and equilibrium enhancer configuration to the mimicked by liquid n-­octanol. (c) permeation of CS across HEM under the sym. asymmetric configurations are likely to be the Thus. being the case because of the lipophilic nature meation experiment findings under the s­ ymmetric of the enhancer site of action. The data show a correlation between with a carbon–carbon double bond does not transdermal permeation enhancement under the significantly affect its intrinsic potency. for skin permeation under the symmetric and cant concentration drop across the SC is observed. and symmetric and asymmetric configurations. this The appropriateness of extrapolating the per. Enhancer B.I. Li and W. This analysis assumes SC is homogenous and does not lead to nonlinear profiles p­ ermeability across the SC. The ation under the asymmetric configurations in relatively constant concentration of Enhancer A practice. potency of the enhancer (as represented by its ined in a recent study (Chantasart and Li 2010). Such enhance- A > Enhancer B. concentration at the target site of action) is In this study. However. 7. Conclusion form transport enhancement over the entire SC New insights into the factors influencing the and a high overall flux enhancement of drug effectiveness of chemical permeation enhanc- transport across SC. view that (a) the potency of an n-alkyl tion of the enhancer in the transdermal patch are enhancer (based on its aqueous concentration) other factors that need to be considered.K. log Koctanol/PBS for Enhancer induced enhancement for the enhancer.134 S. enhancement (permeation and partition enhancement) at Enhancer A. in the SC would suggest that lipophilic enhancers are likely to be more effective in providing uni. a much more signifi. PD/E for Enhancers different locations within the SC and (b) enhancer- A and B are the same.14 Enhancer concentration profiles in SC in account for (a) enhancer-­ induced variation in local transdermal drug delivery (see Eq. a large portion of the SC is not affected by same and supports the utility of the findings in Enhancer B and this region of the SC becomes the symmetric transport studies for skin perme- the rate-limiting barrier for drug transport. membrane Enhancer B Enhancer A Blood sink Combined SC Patch viable epidermis and some dermis Fig. Higuchi Enhancer concentration Ci CD. The solubility of the enhancer and deple. This (d) with modest effects. dashed line.

In: Dermatol 88:702–705 Walters KA. However. the present He N. Higuchi WI (1997) chemical permeation enhancers on the lipoidal path. J Pharm Sci 98:926–944 Abrams K. Harvell JD. Pharm Res 8:865–872 . Higuchi WI. Maibach H. Crit Rev Ther Drug Carrier transport across the stratum corneum. Pugh J. Arch permeation enhancement by chemical agents. Higuchi WI (1992) Model studies Aungst BJ (1989) Structure/effect studies of fatty acid iso. Nevertheless. Pharmaceutics Syst 8:91–192 4:71–92 Li SK. Peck J. This suggests that further studies Hadgraft J. Grubauer G. Prakongpan S. Christophers E (1963) Preparation of iso- Brain KR. Yamamoto A. Higuchi WI.7  Mechanistic Studies of Permeation Enhancers 135 reduces the intrinsic potency of the enhancer. Kevin Li SK (2004) Mechanistic study of chemical skin per- S. Marcel Dekker. we have not encountered any enhancer Lipid content and lipid type as determinants of the epi- dermal permeability barrier. Ghanem AH. Semin Dermatol 11: mers as skin penetration enhancers and skin irritants. J Lipid Res 30:89–96 candidates that are inconsistent with this view Hadgraft J (2001) Modulation of the barrier function of the in our research. J Pharm Sci 93:1415–1430 financial support by NIH Grants GM 043181 and GM He N. J Pharm effects on diffusion and metabolism of beta-estradiol Sci 76:25–28 in hairless mouse skin. sis of flux enhancement across hairless mouse skin induced by chemical permeation enhancers. with hairless mouse skin. Warner KS. Allan G (1996) Mechanisms of action of skin penetration enhancers/ are needed for greater generalization of the retarders: azone and analogues. Wertz P. Sa-Nguandeekul P. Li SK. J Pharm HI (eds) Percutaneous penetration enhancers. 5 Higuchi WI (2007) Comparison of the effects of Li SK. Ghanem A-H. J Pharm ies of chemical permeation enhancers. Li SK. Warner KS.g. In: Dragicevic N. 145–156 Pharm Res 6:244–247 Kligman AM. Ibrahim for their contributions in the project and the functional groups. Int J Pharm 297:9–21 Ibrahim SA. the stratum corneum. Kurihara-Bergstrom T. Effects of long-term hydration leading to the develop- pp 389–416 ment of polar channels in hairless mouse stratum cor- Chantasart D. Hadgraft J (eds) Pharmaceutical skin Lambert WJ. Maibach chain alkanols as skin permeation enhancers. New York. Li SK (2010) Relationship between the neum. Krill SL (1989) penetration enhancement. Warner KS. Walters KA (1993) Molecular modeling of skin lated sheets of human stratum corneum. Chantasart D. Prakongpan S. Lampe MA. Pharm Res 14:S-303 J Pharm Sci 96:2310–2326 Liu P. as skin permeation enhancers by permeation and parti- tion experiments with hairless mouse skin. Li SK (2009a) Effects of chemical enhancers on human epidermal membrane: structure-­enhancement References relationship based on maximum enhancement (Emax). Pharm Res 27:1825–1836 Lee VHL. skin penetration skin. Higuchi WI. 1996). Ibrahim SA. Warner KS. Ning He. Williams ML. McKie JE. Yoneto K. Int J Pharm 141:17–25 present findings. enhancement relationship and the microenvironment of Higuchi WI (2004) Mechanistic studies of branched-­ the enhancer site of action. J Pharm Sci 78:925–928 enhancement effects of chemical permeation enhanc. Knutson K. Shriner D. Shaker DS. Harris RM. Springer. Kompella UB (1991) Mucosal Chantasart D. Higuchi WI. Li SK (2009b) Effects of solvent deposited Maibach HI. penetration enhancers for facilitation of peptide and tionship of chemical penetration enhancers in drug protein drug absorption. chemical Sci 93:762–779 methods in penetration enhancement: modification of Chantasart D. Skin Pharmacol Appl Skin Physiol 14(Suppl 1): retarders have been reported (e. Potts RO (1987) Role of stratum Good WR (1991) Quantitative evaluation of ethanol corneum lipid fluidity in transdermal drug flux. Acknowledgments The authors thank Drs. and Sarah meation enhancers with different polar and lipophilic A. J Lipid Res 24:131–140 in vitro. Warner. He N. Higuchi WI study has demonstrated useful concepts and (2003) Mechanistic study of alkyl azacycloheptanones effective methodologies for mechanistic stud. Elias PM (1989) To date.. of epidermal permeability. Suhonen TM. Ch. Rehfeld SJ (1993) Effect of organic sol. Li SK. Li SK (2012) Structure enhancement rela. Doungdaw Chantasart. Effects of 1-alkyl-2-pyrrolidones on the lipoidal path- ways of human epidermal membrane and hairless way of human epidermal membrane: a comparison mouse skin and the mechanism of enhancer action. enhancers on transdermal permeation and their rela- vents on in vitro human skin water barrier function. New York. Feingold KR. J Control Release 136:117–124 J Invest Dermatol 101:609–613 Kim YH. Li SK (2005) Model analy- 063559. Higuchi WI (2015) Quantitative structure-­ Chantasart D. Hadgraft 72–81 et al. tionship with Emax. Sci 92:297–310 He N. Song WQ. Williams DG. Elias PM (1983) Human epi- ers on the lipoidal transport pathway across human dermal lipids: characterization and modulations dur- skin under the symmetric and asymmetric conditions ing differentiation. Golden GM.

Peck KD. Boca Raton branes. Ghanem AH. group chain length and polar head group on chemical skin dermal transport of low-molecular weight hydropho. Li SK. Science 186:19–26 tionship for chemical skin permeation enhancers: Okamoto H. Int J Pharm 78:123–136 Warner KS. Omar MS. Nicolaides N (1974) Skin lipids: their biochemical Bolikal D. Harris WT pyrrolidones on permeant partitioning into the stratum (1999) Synthesis and evaluation of N-acetylprolinate corneum. Warner KS.I. Maibach HI (1995) Percutaneous penetration permeant size and permeability in lipid bilayer mem- enhancers. Li and W. Li SK. Hao J. Kasina R. Li SK branes. Kurihara-Bergstrom T. Hashida M. Hashem for determination of free 1-alkyl-2-pyrrolidone con- FM. Wiley. PhD Thesis. 2nd edn. CRC Press. J Pharm Sci 90:1143–1153 bic solutes. Anderson BD (1994) The relationship between Smith EW. Xu Q. University of Utah vitro. permeation enhancement. Yoneto K. J Pharm Sci transdermal permeation of corticosterone through 97:368–380 hairless mouse skin. Higuchi WI (2001) Influences of alkyl Mitragotri S (2001) Effect of bilayer disruption on trans. J Pharm Sci 92:1305–1322 derivatives as percutaneous penetration enhancers. Warner KS. Shimabayashi S (1998) New York Influence of the permeation enhancers 1-alkyl-2-­ Tenjarla SN. J Membr Biol 140:111–122 Stein W (1986) Transport and diffusion across cell mem. Ghanem AH. Academic. Li SK. Ghanem A-H. Int J Pharm 253:1–11 Xiang TX. Li SK (2008) Silicone elastomer uptake method Shaker DS. J Pharm Sci 84:312–317 micelles and biological membranes. Chantasart D. Suhonen TM. Higuchi WI. Shaker DS. Li SK. Higuchi WI. Higuchi Liu P. J Pharm Sci 87:209–214 .136 S. Int J Pharm Good WR (1992) Assessing the influence of ethanol in 192:147–158 simultaneous diffusion and metabolism of estradiol in Warner KS (2003) Mechanistic aspects of chemical skin hairless mouse skin for the ‘asymmetric’ situation in permeation enhancers.K. New York (1995) Mechanistic studies of the 1-alkyl-2-­pyrrolidones Tanford C (1980) The hydrophobic effect: formation of as skin permeation enhancers. Molokhia S. Yoneto K.. Higuchi WI (2003) Structure-activity rela- uniqueness. Higuchi WI. Sezaki H (1988) Structure-­activity probing the chemical microenvironment of the site of relationship of 1-alkyl or 1-­alkenylazacycloalkanone action. esters – novel skin penetration enhancers. Higuchi WI (2003) Mechanistic studies of the centration in micelle and hydroxypropyl-β-­cyclodextrin effect of hydroxypropyl-beta-cyclodextrin on in vitro systems used in skin transport studies. Puranajoti P. Higuchi J Pharm Sci 77:418–424 WI. Pharm Res 18:1018–1023 Warner KS. He N. Inc.

..1 Skin Impedance–Skin Permeability old as human civilization... 137 The idea of delivering drugs through the skin is as 8....... 140 8............ This barrier must be altered to maximize the advantages of transdermal route of drug administration... and thermal poration Troy............ USA duction of a variety of techniques including for- e-mail: ajain13@gmail. 146 term delivery (>24 h).... NY............. long- Understanding... Skin’s topmost layer. H....).. 144 transdermal route of drug administration offers 8...... USA (Sintov et al....4 Applications of INSIGHT Screening...........4.. © Springer-Verlag Berlin Heidelberg 2017 137 N.. iontophoresis (Kalia et al. Jain (*) • S.. and Biological Engineering. and Samir Mitragotri Contents 8.. 145 several advantages such as reduced first-­pass drug 8....... Menlo Park...2 Generation of Database for Quantitative metabolism... Rensselaer Polytechnic Institute...... Maibach (eds. 2003)...... jet injection Center for Biotechnology and Interdisciplinary Sciences... USA acoustical methods (Mitragotri and Kost 2004)......... Rensselaer Polytechnic Institute. (Hingson and Figge 1952)....... dermatolo- gists.. only few drug molecules have been formulated into transdermal patches (Barry 2001). This has engaged pharmaceutical scientists...2 Overview of INSIGHT Screening......2. and control over delivery References..... 145 8. NY.. Though 8......1007/978-3-662-53270-6_8 ..... 1999).. Methods.. microneedles (Prausnitz 2004)......3 Validation of INSIGHT with FDC...............I. but the excitement has Correlation.. High A..........1 Discovery of Rare Formulations.............. Mitragotri research activity in this field has led to the intro- Corium International lnc........... Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin.. Dragicevic......... stratum corneum (SC)..com mulation-based approaches (Williams and Barry P....... DOI 10. CA.. Pankaj Karande.... Weaver et al...... no gastrointestinal degradation....... electro- Howard Isermann Department of Chemical poration (Prausnitz 1999.. 147 and termination..4.1 Introduction. and engineers alike in research over the last couple of decades (Mitragotri 2004)...........1 Introduction 8............ 141 increased in recent times after the introduction of the first transdermal patch in the 1970s... High Throughput Screening of Transdermal Penetration 8 Enhancers: Opportunities. forms a barrier against permeation of xenobiotics into the body and water evaporation out of the body.. Karande 2004). Troy....... The cause of this imbalance between the benefits of this route and the number of products in the market lies in the skin itself... and Applications Amit Jain......... 2004).

1982. Starting with a pool of >200 individual based approaches have a number of unique CPEs. these methods are to screening the potency of enhancer combina. Formulation-­ approach. However. Accordingly. 2000). and ease of application over a large area these mixtures is a mammoth task. Thomas and flow through diffusion cells has been developed Panchagnula 2003). 1989). anionic surfactants et al. dermal drug delivery. All of the above techniques have their own one has to rely on a brute force screening benefits and specific applications. Occurrence of truly synergistic combinations stages. To date. to use CPEs as building 1996. which form the core component of ments are likely to yield better results. These include et al. enhancers has proved challenging. order formulations can be designed. also cost prohibitive. 2000). The last 20 years have Screening of chemical enhancers can be per- seen extensive research in the field of chemical formed in vitro as well as in vivo. 2004). Ogiso et al. 2003). Guy tants (Lopez et al. The use of in vitro and Barry 1991. Furthermore. Franz diffusion cells (FDCs). the novel formulations that offer enhancement with. the challenge now shifts cantly improved. Jain (Priborsky and Muhlbachova 1990). exhibit additive properties. 1987. standard tions. tions. Automated in-line constituents (Mitragotri 2000. throughput screening methods. and methyl dermal drug transport have been performed using laurate among others. millions of binary and billions of higher advantages such as design simplicity and flexi. shows similar behavior in vivo and in vitro except only few chemicals are to be found in currently for the extent of metabolic activity (Chang marketed transdermal products. 2000). Random mixtures of CPEs are likely to FDCs still dominate the screening of CPEs. Jain et al. (HPLC) or radiolabeled drugs for liquid scintilla- mal drug transport compared to their individual tion counting are expensive. However. 2002). (Prausnitz et al. 1990. based on in vitro data (Naito and Tsai 1981. Accordingly. nonionic surfac. and drug delivery. Takayama and (Nokhodchi et al. the principle site of enhancer action. More than 200 chemicals have practicality limit their applications for screening been shown to enhance skin permeability to a large database of enhancers. Screening of bility. however. various drugs. fatty esters models exist to predict in vivo pharmacokinetics (Chukwumerije et al. models for screening is also supported by the fact identification of potent yet safe permeation that SC. throughput has led to the development of high sponding properties of their individual constitu. 1989. cost. be feasible. These include molecules from a in vitro screening based on excised tissue (human diverse group of chemicals including fatty acids or animal) presents a more practical alternative (Golden et al. A high throughput assay to . Although these blocks to construct new microstructures and methods have facilitated the experiments. that is. combinations of CPEs represent a huge These permeation studies are time consuming opportunity that has been sparsely tapped. and terpenes (Williams Nagai 1991.138 A. Though in early ents. In the absence of capabilities ise in discovering novel formulations for trans- to predict the occurrence of such rare mixtures. It should skin permeation experiments (Bosman et al. their potency The urgent need to increase experimental and irritancy are likely to be averages of corre. and are resource expensive as analytical methods Several reports have indicated that combinations such as high-pressure liquid chromatography of CPEs offer better enhancements of transder. sorbitan monooleate. low. However. formulation-based strategies for transdermal several issues including variability. In vivo experi- enhancers. Aungst et al. Cordoba-Diaz et al. throughput of these methods has not been signifi- out irritation. A number of and Panchagnula 2003). such combinations in the last few years to increase the throughput of do not necessarily yield safer enhancers. not more than 10–15 experiments at a time. these methods have already shown prom- is likely to be rare. Wu et al. in principle. 1998). Jain et al. Majority of in vitro studies on trans- oleic acid. The throughput of Even though individual chemical penetration this traditional setup of diffusion chamber is very enhancers (CPEs) have found limited applica.

beled (Rosado et al. Permeation enhancement efficacy of a Takács-Novak and Szász 1999. the high throughput screening Permeation experiments using radiola. a model solute across the skin in the pres. CPE is a function of its chemical potential 2003a. FDCs typically require application of tions should meet the following requirements: 1–2 ml of enhancer formulations over about 3–4 cm2. frequencies for assay utilization. 2001). should be able to decouple these two effects Narishetty and Panchagnula 2004). ence of an enhancer is dependent not only 2003a. pig skin (Sekkat et al. This makes (i) Ability to screen a large number of it cost prohibitive to include candidates that formulations are expensive in the test libraries as well as Increasing the throughput by at least 2–3 to screen a large number of formulations. tions need to be standardized for all the tion times of 48–96 h thereby reducing the enhancers that are being tested to create throughput of permeation experiments. temperature (Ongpipattanakul et al. Furthermore. if possible. fluorescent the following experimental constraints: (Ogiso et al. direct comparison of their efficacies in Low incubation times favor high turnover increasing skin permeation. show deviations in permeability characteris- ples handled in a given time. 1996). 1990). excised efficacy of formulations. effect of an enhancer on skin permeability. 2001) among other thermodynamic the throughput and hence time efficiency parameters. Auner et al. Larrucea (iii) Low incubation times to further increase et al. or RIA/ELISA-detectable resent human skin (Xing et al. Also. 2001). (iv) Minimal use of test chemicals and efficient This chapter focuses on a specific high utilization of model membrane such as ani. Magnusson and Runn It is common to find in transdermal litera- 1999) markers necessitate the need of exten. pling with little opportunities for process (ii) Use of a surrogate end point that is quick. b). methodology should also satisfy. easy and independent of the physicochemi. throughput screening method called INSIGHT. 1991. (vii) Use of consistent thermodynamic condi- chemical interactions of the enhancer with tions for enhancer formulations the model solute (Lee and Kim 1987. cosolvent (Yamane et al. and to assure the generality of the results. These thermodynamic condi- FDC experiments typically use incuba. human skin. An end point. HPLC-detectable (Wu (vi) Use of a common model membrane to rep- et al.8  High Throughput Screening of Transdermal Penetration Enhancers 139 be used for screening of transdermal formula. (v) Adaptability to automation to reduce human cant reduction in the effort and time spent interference in the very first stage of formulation devel. Auner et al. In addition to these requirements of the cal properties of the model permeant assay tool. 2000). represent human skin such as rat skin This accentuates the cost of sample analysis (Schmook et al. snake skin (Itoh et al. 1997. to characterize the (Francoeur et al. 2000). Typical FDC setup requires manual sam- opment (Karande and Mitragotri 2002). Shokri et al. 1998. 2001. orders of magnitude would result in signifi. ­animal models a fundamental limit on the number of sam. etc. permeabilize skin but also on the physico. and overall time spent in characterizing the 2002). Permeation of tics from human skin (Panchagnula et al. automation (Cordoba-Diaz et al. 1995. results on one model cannot on the inherent capacity of the enhancer to be directly translated to a different model. of skin per experiment. While human skin is ­difficult ­current state-of-the-art fluidics systems put to procure on a large scale. mal skin IN vitro Skin Impedance Guided High . ture the use of a variety of different models to sive sample handling and sample analysis. 2003). 1990. Schmook et al. b).

2  verview of INSIGHT O INSIGHT screening uses skin impedance as a Screening surrogate marker for skin permeability. in its current version. Skin impedance has been previously used: (i) INSIGHT screening offers >100-fold improve.1). the duced (Karande et al. can perform up described in detail with respect to its fundamen. 2004). Second.140 A. to 50 tests per square inch of skin compared to tals. The skin is sandwiched moved sequentially in the donor wells. 8. Conductivity measurements are made with one a symmetric receiver array (bottom). This improvement in et al. 8. efficiency comes from two factors. First. a b Electrode Donor Skin Receiver ~ Electrode Fig. A single screen can electrode inserted in the dermis and a second electrode screen 100 formulations at one time. to assess the skin integrity for in vitro dermal test- ment in screening rates of transdermal formula. about >2 cm2 of skin per test in the case of Franz diffusion cells (Fig. 8. and the formulations contact the stratum corneum from the The INSIGHT screen is made up of a donor array (top) and donor array. About 100 formulations can be screened per INSIGHT array. ing (Lawrence 1997. respectively Throughput screening that was recently intro. Jain et al. (a) and (b) is top and between the donor (Teflon) and receiver (Polycarbonate) side view of the INSIGHT apparatus. 2004). This method is INSIGHT. Davies tions (Karande et al. 2004). Fasano et al. and outcomes. 2002. validation. (ii) to evaluate the irritation potential .1  Schematic of the INSIGHT screening apparatus.

2002).N-dimethyl-3-ammonio-1-propanesulfonate). OL. lauric Yamamoto 1976a. electrical impedance and solute permeability is tion test (SIFT) (Heylings et al. Data reported in this chap. S20. between skin conductivity and skin permeability. quencies facilitates measurements due to SO. SLS. Thus. and others . Therefore existing data cannot be used to gener- ers and their validity for a wide range of markers alize the relationship between skin impedance have not been previously documented. fatty acid esters (TET. oleic acid. BDAC. cetyl tri- potential across the skin or by measuring tran. chosen from the candi- lipids in which protein-rich corneocytes are sur. acid. electric and Yamamoto 1976a. an optimal monitor skin barrier recovery in vivo following the frequency must be chosen. polyethylene the capacitive components of the skin. tetra- decreased impedance. Scattered literature data impedance (conductivity) and permeability rela- support this hypothesis. benzyl sepidermal current following the application of a dodecyl ammonium chloride). Approximately 70–100 bilayers are stacked The candidate pool comprised of 15 single between two corneocytes (Elias et al. sodium oleate). IPM. Curdy et al. quantitative current (Sims et al. zwitterionic surfactants (HPS. with increasing frequency (Yamamoto and fatty acids and their sodium salts (LA. the correlation between caine HCl. A study by Yamamoto tionship. Accordingly. SOS. and permeability. SLA. b). sorbitan monolaurate. 1997. A set of 22 Stratum corneum is a composite of proteins and enhancer formulations. it is logical to hypothe. 1999). the mea. date pool was used to validate the relationship rounded by lipid bilayers (Madison et al. tants (NLS. ter are based on measurement of transepidermal sodium laureth sulfate. 2002). Since it is evident quency of 100 Hz. Because of its architecture the SC is rela. and moreover in most of the studies this and Yamamoto (1976a. sodium octyl sulfate. formulations. Elias enhancer formulations and seven binary enhancer 1983).8  High Throughput Screening of Transdermal Penetration Enhancers 141 of chemicals in a test known as skin integrity func. b) showed that total skin relationship was used to elucidate the mechanism impedance reduces gradually with tape stripping of transport of hydrophilic molecules across the and after 15 stripping skin impedance approaches skin under the influence of temperature (Peck the impedance value of deep tissues (Yamamoto et al. anionic surfac- constant AC potential. 1991. polyoxyethylene sorbitan monolaurate). a large dataset was first generated to assess the correlation between skin impedance and permeability to 8. methyl ammonium bromide. Tezel et al. isopropyl myristate). Due to nonionic surfactants (PEGE. sured electrical impedance of the skin decreases T20. and (iii) to stronger at lower frequencies. While the use of higher fre. from the literature that skin impedance can be used INSIGHT screening is founded on the rela- to confirm skin integrity. 1998). 1995). potential is also an important parameter. Skin wide range of chemical enhancers. 1977. b). sodium lauryl sul- current following the application of a constant fate). N-lauorylsarcosine sodium. There is a dearth of literature on skin marker for permeability. 2001. 1987). In order to establish the correlation tively nonconductive and possesses high electri. 2003). However. between skin impedance and permeability for cal impedance (Lackermeier et al. Frequency of the applied N. ­permeability in the presence of chemical enhanc.2. All experiments application of current during iontophoresis (Turner reported in chapter were performed at a fre- et al. hydration (Tang et al. tionship between skin’s electrical impedance size that alteration in skin barrier due to chemical (reciprocal of skin conductance) and solute per- enhancers can be used as an in vitro surrogate meability. dodecyl glycol ether. 2001). formulations impedance (AC) can be measured either by were made from different classes of chemicals applying a constant current and measuring the including cationic surfactants (CTAB.1 Skin Impedance–Skin small (mannitol) and macromolecule (inulin) Permeability Correlation hydrophilic solutes in the presence of different chemical enhancer formulations. N-hexadecyl-­ potential (100 mV rms). or ultra- relationships between skin impedance and sonic waves (Tang et al. Li et al.

mannitol r2 = 0.5 %w/v).0  %w/v.0 %w/v).2  Skin impedance–permeability correlation for (a) inu.  -NLS:S20 (1. -PEGE (1. 0. -S20 (2.1) was used as a representa. mals and anatomical regions.85) and mannitol (with OL tive of small hydrophilic drugs.3a. -DMP:HPS (1.  -TET (1. -CTAB parenthesis total concentration of chemical enhancer w/v. logKo/w −3. Permeability data of mannitol There is a reasonable scatter in these data.0 %w/v). -LA (1.4:0. were mea. (b) -OL (1.5).0 %w/v.4). 8. 8. b).6).5 %w/v).4). men. -DMP (1. -S20 (1.6). -TET:SLS (1. -DMP:TET (2. -BDAC (1.5 %w/v).6:0.5 %w/v).4).  -HPS (1.5 %w/v). The measurements over ~5 kΩ-cm2 intervals (inulin r2 = 0. - 0.5 %w/v).5 %w/v).4). (1.0 %w/v). (1. -DMP (2. reported in Fig. 142 A.2.0 %w/v).0-%w/v).2a. 0. a b 4 10-4 10-3 Inulin skin permeability (cm/hr) Mannitol permeability (cm/hr) 10-4 10-4 7 10-5 10-5 4 10-5 10-6 10 100 1 10 100 Skin impedance (kΩ-cm2) Skin impedance (kΩ-cm2) Fig. -OL (1.0 %w/v). -MEN:T20 (2. which and inulin with a variety of chemical enhancer is inherent to biological systems such as skin that formulations showed that skin impedance is exhibits high variability. measurements inversely related to permeability of hydrophilic .2a. Mannitol (MW were plotted separately. mannitol and inulin.2 Da. Inulin (MW 5 kDa) was selected as a between skin permeability and impedance was model solute as it satisfactorily represents a mac.4:0.4). 0. -NLS (1. 8.4:0.9). improved when data for individual enhancers romolecular hydrophilic drug.5 %w/v). N-dodecyl 2-pyrrolidone. DMP enhancer r2 = 0.5 %w/v).0 %w/v).5 %w/v). -OL:MEN (1.0 %w/v.86 and for reported in Fig. Skin impedance and permeability to two experiments performed over several different ani- model solutes. 0. b are replotted after averaging (Figs.5 %w/v. The correlation sured.6:0. (DMP. Also. The A strong correlation was observed between correlation between skin permeability and imped- skin impedance and permeability of mannitol and ance can be clearly seen in Fig.0 %w/v. 8. lin and (b) mannitol. b where inulin for different enhancer formulations data in Fig. weight fraction used): (a) -MEN (1.5 %w/v). -TET:HPS (2. 8. example for inulin (with 182. Test formulations used in this study (in 0. -SOS (2. .0  %w/v. MEN.1:0. (1.6:0. 8. Jain et al. -NLS (1.4a.5 %w/v) NLS:S20 (1.3. -TET (2.0 %w/v. -IPM (1. -PEGE (1. -SO -DMP:TET (2. 0. 0.5 %w/v). b represent an aggregate of thol).0 %w/v).90). and 8.5 %w/v). -LA MEN (1.0 %w/v. b were performed in FDCs. 8.6:0.0 %w/v). enhancer r2 = 0.2a. -DMP (1. 0.5 %w/v.86) is given in (Fig.6).5:0.  8.

8. 8  High Throughput Screening of Transdermal Penetration Enhancers 143 10-3 a 5 10-4 b Mannitol skin permeability (cm/hr) Inulin skin permeability (cm/hr) 3 10-4 10-4 10-4 8 10-5 6 10-5 2 4 6 8 10 5 6 7 8 9 10 20 30 Skin impedance (kΩ-cm2) Skin impedance (kΩ-cm2) Fig. 8. Permeability data for lation is much tighter as compared to the one in Fig. EtOH:PBS). 8.5%w/v in 1:1 single enhancer.2 EtOH:PBS).2. 8. 10-3 a b 4 10-4 Mannitol skin permeability (cm/hr) 3 10-4 Inulin skin permeability (cm/hr) 10-4 2 10-4 10-4 10-5 9 10-5 8 10-5 7 10-5 6 10-5 5 10-5 10-6 10 100 1 10 100 Skin impedance (kΩ-cm2) Skin impedance (kΩ-cm2) Fig. (a) Plot of skin permeability to inulin vs. The corre- impedance data shown in Fig.4  Skin impedance – permeability correlation for different enhancers are grouped in the bins of 5 kΩ-cm2 (a) inulin and (b) mannitol. Modified plot of permeability-­ along the x-axis representing skin impedance. 8. (b) Plot of skin permeability to mannitol vs.2 .3 Skin impedance-permeability correlation for skin impedance in the presence of NLS (1.5%w/v in 1:1 compared to Fig. A much tighter correlation can be observed skin impedance in the presence of DMP (1.

 Plot of conductivity enhancement ratios in circles indicate permeability enhancement numbers in INSIGHT at 24 h vs. ratio of skin impedances at time zero and 24 h meability and impedance for single and binary following the application of enhancer formula- enhancers formulations irrespective of the nature tion. >25-fold reduc- physicochemical properties of the drug or tion in skin utilization). and the filled FDC. however. in assessing formulation potency after a 4-h 100 45 1000 Formulation rank based on 4 h Incubation 40 80 FDC permeability enhancement FDC conductivity enhancement 35 30 100 60 25 40 20 10 15 20 10 0 5 1 0 10 20 30 40 50 60 70 1 10 100 1000 INSIGHT conductivity ER at 24 h Formulation rank based on 24 h Incubation A B Fig. Correlation coefficient (inulin with FDC r2 = 0. INSIGHT enhancers. Results shown in Fig. is not precisely known).90) of average data for all enhancer formulations indicates that a Conductivity enhancement ratio (ER). dictions in INSIGHT on the potency of enhancer formula- lations. tions obtained from INSIGHT on the potency of Specifically. A strong linear correlation indicates the validity of tions. Note. IPM).5 Validation of INSIGHT predictions with cate conductivity enhancement numbers. measurements in INSIGHT were plotted of the formulation. that is. (a) Plot of 24 h predictions in INSIGHT vs. was used as a model permeant in these studies. good correlations were observed enhancer formulations are essentially the same as between permeability and skin impedance for those obtained from FDCs. lower incubation periods of 4 h . allows collection of information at a much greater MEN. Educated discretion must therefore be Further improvements in INSIGHT screening exercised when selecting a delivery formulation speed can be obtained by reducing the formula- for a particular model permeant or drug of tion incubation period. solutes.3  alidation of INSIGHT V in literature.07 cm2 per experiment as compared to these correlations is an integral function of the 2 cm2 in a 16 mm diameter FDC. 8. Capabilities of INSIGHT interest. The closed circles indi. that the nature of (about 0.5a reflect that the predic- in most cases. These results indicate that against conductivity enhancement and permea- skin impedance can be used as a parameter to bility enhancements in FDCs (Fig. 144 A. 8. 4 h pre- enhancement ratios in FDC at 96 h for 19 enhancer formu. cals irrespective of their mode of action (which. which act by lipid extraction (NLS. However. LA. Jain et al. A strong correlation indicates that predictions on observations in INSIGHT when compared with those potency of formulations can be obtained at significantly from traditional tools like FDC. which is in agreement with existing data 8. Inulin measure the extent of barrier alteration by chemi. conductivity and permeability FDC.86 and mannitol r2 = 0. 8. the remarkable correlation exists between skin per. ­permeant.5a). BDAC) or by lipid bilayer fluidization speed (~1000 per day) and less skin utilization (OL.

the INSIGHT screening can be further improved. ated enhancer combinations that exhibit ER tional tools.1  % of formulations novel chemical enhancers such as laurocapram exhibited more than 60-fold enhancement of skin (Azone). tions by brute force experimentation is contin- cially for macromolecules. Random pairing of CPEs from various cate- library based on 4-h screening are compared to gories led to 496 binary chemical enhancers those based on 24-h screening. achieving sufficient potency conductivity. About 20 % of this library (5. the same task was Formulations accomplished in about 2 months with screening rate of 500–1. INSIGHT screening can be used to screen huge Binary formulations exhibited a wide range of libraries of chemicals within a short span of time enhancements.000 experiments per day.4. Animal from the first principles is challenging. increasing threshold (Fig. INSIGHT screening opens up the possibility of tion of chemical enhancers are more potent than its discovering such rare formulations. 2004). 2002. The percent of randomly gener- and without the fear of failure that exists with tradi. enhance skin permeability. using a SCOPE formulation (SLA:PP) is signifi- tration enhancers (SCOPE) formulations. Using the traditional Screening tools for formulation screening. Less than 0. 2004. However. Tezel One of the formulations discovered by INSIGHT. SLA:PP (sodium laureth sulfate:phenyl piperazine) ponents increases the number of formulations was shown to increase the permeability of macro- exponentially. Prediction of synergies and oligonucleotides by 50–100 fold. Many current single enhancers are also above a certain threshold decreases rapidly with potent irritants to the skin at concentrations neces. pairs. it would have taken over 7 years to do these many experiments. A number of studies gent on the throughput of the experimental tool.5b where from 32 chemicals chosen from a list of >250 potency rankings of 438 single and binary formu. 8. yielding a library of 25. offer insight into mechanisms by which CPEs luteinizing hormone releasing hormone (LHRH). the use of INSIGHT molecules such as inulin across porcine skin by screening allows one to tackle this challenge in a 80–100 fold compared to passive skin permeability more cost-effective way compared to FDCs. leuprolide acetate synergies (positive or negative) between the CPEs. The inset shows sary to induce meaningful penetration enhance. espe. 8. In of inulin (Karande et al.000 experi- 8. synergies between CPEs not only lead to increased the skin permeability of molecules such new transdermal formulations but also potentially as methotrexate. a section of the main figure corresponding to high ment. The amount of leuprolide acetate delivered To identify synergistic combinations of pene. a cantly higher than that delivered from a control library of chemical enhancers was first generated solution and lies in the therapeutic window. et al. SLA:PP also addition. have shown that formulations made up of combina. INSIGHT experiments in hairless rats also confirmed delivery screening offers an effective tool for identifying of a synthetic analogue of LHRH. thus indicating that the efficiency of formulations) was screened using INSIGHT. however. Rank 1 corre. Discovery of such rare formula- without irritancy has proved challenging. 44 distinct chemical compo- sponds to most potent formulation in the library sitions were created with the concentration of and rank 438 to the weakest formulation. Mitragotri 2000).040 24 h.6a). predictions of the potency made in 4 h were con. The addition of com. The each chemical enhancer ranging from 0–2 % w/v. For each pair. largest ever-cohesive screening study reported in the transdermal literature. Attempts have been made to synthesize ER values. individual components (Karande et al. Each formulation was tested at least four times in over 20.000 candidate SCOPE sistent with those made after a contact time of formulations.1 D  iscovery of Rare With INSIGHT screening. . 8.8  High Throughput Screening of Transdermal Penetration Enhancers 145 i­ncubation are demonstrated in Fig. in vivo.4  pplications of INSIGHT A ments (Karande et al. chemical enhancers belonging to various catego- lations randomly prepared from the enhancer ries. 2004). low molecular weight heparin.

can be used to generate large volumes of data on the interac- ing these potent formulations is very small (~0. Regions of high synergy almost always overlap to discover using the traditional tools like FDC due to their with the regions of high potency.1 %) requir. tion generated via INSIGHT screening on the 2003).146 A. (a) Discovery screening is used to quantify the extent of interactions of rare enhancer formulations that is significantly potent in between the components of CPE mixtures in terms of increasing skin permeability. for the first time. 8. which have had limited suc- tions of enhancers. The information is used to relate ing a tool with high experimental throughput. (QSARs) for CPEs. (b) INSIGHT chemistry of the enhancer to its potency using QSAR 8. Database for QSAR Fig. availability of large volumes of data collected in a . (c) INSIGHT screening low experimental throughput.01% 30 40 50 60 Conductivity enhancement ratio 10% 5% 0 0% 0 1. The success rate of discover. cess in the past (Moss et al. a.(%w/v) 1% 25% 20% 0. Discovery of rare formulations b.2 G  eneration of Database ers will provide. the sheer volume of informa. Exploration of synergy 35% 2.1% 15% 0. Quantitative descriptions of structure-activity relations Looking beyond searching for potent combina.6  Applications of INSIGHT screening.0 Percent of enhancer formulations 10% 30% Percent of enhancer formulations Total conc. tion of CPEs with skin. a platform to for Quantitative build further investigations of the fundamental Understanding aspects of enhancer-skin interactions. Walker et al. 2002. Jain et al. may lead to better outcomes in light of the behavior of a wide variety of penetration enhanc.0 0 10 20 30 40 50 60 Fraction of MP Conductivity enhancement ratio A B Insight C Building predictive capabilities INSIGHT Bulk Discrete observable structural parametrical behavioral QSAR Relating activity to structure c. Such formulations are difficult Synergy.4.

Mitragotri S (2002) High throughput screen- automated dynamic sampling. this Elias PM. ment for in vitro permeation studies. Blake JA et al (1990) Contributions of drug 1042–1047 solubilization. enable successful transdermal drug delivery. penetration of tenoxicam and its retention in the skin. I. Adv Drug Deliv Rev 56(5):619–658 for in vitro transdermal permeation. Pharm Res 19(5): 14(8–10):1015–1023 655–660 Chang P. Also. barrier disruption. As exemplified in Fig. Kim SW (1987) Hydrophobization of ionic drugs Davies DJ. compatible with Karande P. Francoeur ML. this knowledge Golden GM. Kalia YN et al (2002) Post-iontophoresis recov. Toxicol In Vitro 11:241–249 Biopharm 53(1):15–21 Lee SJ. Pharm Res 7(7):712–718 Jain AK. ation of various compounds with fatty acids and Methods Find Exp Clin Pharmacol 25(6):413–421 amines. Ward RJ et al (2004) Multi-species assessment for transport through membranes. 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Toxicol In Vitro 16(3):299–317 quaternary ammonium drugs: the influence of counter Naito SI. Cross SE et al (2003) Effect of vehicle pretreat. Int J Pharm ing hormone analogue pGlu-3-methyl-His-Pro amide 228(1–2):99–107 through human epidermis. Mitragotri S et al (2001) Theoretical description effect of surfactants on the penetration of lorazepam of transdermal transport of hydrophilic permeants: through rat skin. Rodford R et al (2003) Quantitative structure-­ Prausnitz MR. Nokhodchi A et al (2001) The effect of surfac- enhancers on the permeation of the thyrotropin releas. Nat Rev absorption rates. Methods Find Exp Clin transport of hydrophilic solutes during low-frequency Pharmacol 19(5):335–341 sonophoresis based on a modified porous pathway Peck KD. Higuchi WI et al (1991) Skin alteration and Mitragotri S (2000) Synergistic effect of enhancers for trans. Int J Pharm 250(2):359–369 application to low-frequency sonophoresis. Jain et al.148 A. Meingassner JG et al (2001) Comparison of effects of Span20 with Tween20 and Azone on the human skin or epidermis models with human and ani- in vitro percutaneous penetration of compounds with mal skin in in-vitro percutaneous absorption. Adv Drug J Pharm 69(2):109–121 Deliv Rev 56(5):555–556 Sintov AC. Int Mitragotri S (2004) Breaking the skin barrier. Stemmer K et al (1997) Animal models Tezel A. J Pharm Sci 91(11):2376–2381 Magnusson BM. Burnette RR et al (1991) Evidence Tezel A. Swartzendruber DC et al (1987) Presence of Sekkat N. Int different lipophilicities. and flexibility. convective solvent flow effects during iontophoresis: dermal drug delivery. Int J Pharm 202(1–2):133–140 J Pharm 215(1–2):51–56 Madison KC. Pharm Res 20(9):1502–1507 enhancement. Muhlbachova E (1990) Evaluation of in-vitro Weaver JC. Vaughan TE et al (1999) Theory of electrical percutaneous absorption across human skin and in ani. absorption and skin damage of ketoprofen hydrogels nism of action. J Invest Dermatol 88:714–718 in vivo. Turner NG. J Pharm Sci 85(1):57–64 91(8):1891–1907 Ongpipattanakul B. Shokri J et al (2003) The enhancement Tang H. Nagai T (1991) Simultaneous optimization Narishetty ST. retention. Adv Drug Deliv Rev 35(1):21–39 Rosado C. creation of aqueous pathways across skin transport mal models. indomethacin from ointment bases in rabbits. J Pharm Ogiso T. model. Szász G (1999) Ion-pair partition of absorption. J Pharm Pharmacol 42(7):468–472 barriers. Niinaka N et al (1996) Mechanism for enhance. Kalia YN et al (2002) Biophysical study of por- intact intercellular lipid lamellae in the upper layers of cine ear skin in vitro and its comparison to human skin the stratum corneum. Int J Pharm 74:115–126 Nokhodchi A. Environ Toxicol Chem 22(8): Drug Discov 3(2):115–124 1870–1884 Priborsky J. Pharm Res 17(11):1354–1359 I. J Pharm Sci absorption. Pharm Res 8(1):17–24 . Tsai YH (1981) Percutaneous absorption of ions of different lipophilicity. Lopez A.

Int J Pharm 14(2):151–158 209(1–2):87–94 Yamane MA. J Control Release 52(1–2):89–98 J Pharm Pharmacol 47(12A):978–989 . Lin S et al (1998) Transdermal testosterone tion enhancers in propylene glycol/water co-solvent delivery in castrated Yucatan minipigs: pharmacokinet. Yamamoto T. Yamamoto Y (1976a) Dielectric constant Adv Drug Deliv Rev 56(5):603–618 and resistivity of epidermal stratum corneum. Yamamoto Y (1976b) Electrical properties transdermal hydrophilic gels in normotensive rabbits of the epidermal stratum corneum. Huang YB et al (2000) Evaluation of pharmaco. ics and metabolism. Barry BW (2004) Penetration enhancers. Med Biol Eng and spontaneously hypertensive rats. Williams AC et al (1995) Terpene penetra- Xing QF. Med Wu PC. Biol Eng 14(5):494–500 kinetics and pharmacodynamics of captopril from Yamamoto T.8  High Throughput Screening of Transdermal Penetration Enhancers 149 Williams AC. systems: effectiveness and mechanism of action.

Part II Methods for Measuring the Percutaneous Drug Penetration .

...... NY 12180....... Rensselaer accelerated advances in understanding protein Polytechnic Institute.......................... 154 9...................... H............. 159 Conclusion......1 Transdermal Drug Delivery....2 Excised Animal Skin.. Dragicevic.......1 Introduction............... Oral delivery is by far the Environmental Chemistry Laboratory................. Maibach (eds..... 2902 Newmark Dr.. 170 9............3 Proteins of Stratum Corneum....... 160 9............ 161 9.....................2............2 Scope of Review.......).......6...army.......2...4.. DOI 10......4 Polymers... This Development Center.......... 158 9.........3...................mil tide-based drugs sensitive to enzymatic degrada- P.....M.3. 159 9.......2 Pharmacological Response......................5................................ Army Corps drugs especially when repeated or routine admin- of Engineers......2 Intracellular Route....................5.............. Cropek (*) mucosal formulations.............1007/978-3-662-53270-6_9 .........2 Tape Stripping................ 163 9.... 164 9.... 156 9.3.......... Karande (*) tion in the gastrointestinal tract............................................5  valuation of Skin Permeability E in Vitro................................. 158 9...........................................Cropek@usace...... 172 9.......... 167 9...I........ and Measurements in Transdermal 9 Drug Delivery Donald M...............4.........edu bygone drug delivery systems have been domi- © Springer-Verlag Berlin Heidelberg 2017 153 N...... Percutaneous Penetration Enhancers Drug Penetration Into/Through the Skin.......... 164 9............5..... 170 9......... is offset for protein and pep- e-mail: Donald....................... 162 9........5 Lipids ... IL 61822.. USA chemistry and drug interactions............ USA advantage... Engineering Research and istration is required (Chen and Langer 1998).............1........... primarily due to and Interdisciplinary Sciences.. 170 9....................... 169 9... 167 9.....6........... Models....... Construction easiest and most convenient way of delivering Engineering Research Laboratory... Drugs based on Howard Isermann Department of Chemical and proteins and peptides now form a significant frac- Biological Engineering....5 Trans Epidermal Water Loss............ while the e-mail: karanp@rpi..... Cropek and Pankaj Karande Contents 9.....M......................1 Introduction Commonly employed delivery systems include injections............................1 Excised Human skin....... Troy....4...................... U...... 155 9...........................S.4..5............ Champaign...6  valuation of Skin Permeability E 9...........5.........3 Intercellular Route.. 153 9...... 170 9...1 Diffusion Measurements... pills............6.....................3 Impedance Spectroscopy.....3 Other Approaches.......2...3 Routes of Permeation.. 157 9......2 Lipids of Stratum Corneum....... Thus.....4.......1.4 In Vitro Skin Models.. 172 9................1 Diffusion Measurements....1 Ultra-structure of Stratum Corneum...4 Infrared Spectroscopy................................ 157 in Vivo............................. however............. 160 References. 3217...... 165 9.... 168 9....... 155 9.....2 Structure of Skin...................................... Rm...................1 Skin Appendages...3 Living Skin Equivalents... Methods........... Center for Biotechnology tion of the therapeutic spectrum..... 110 8th St. and to some extent topical and D.........................

Miller to its role as a barrier.5) and have molecular It is thus sufficiently obvious that as we move weight (MW) less than 500 Da (Bos and Meinardi toward the next era of health care. Karande nated by the oral route. 2000). both physical and biologi- and Pisani 1999). as a port of entry for systemic delivery of drug Several technological advances have been molecules (Guy 1996. trans. needle-based drug administration has several skin naturally offers a very low permeability to limitations. log P >1. which is results in rapid clearance of active drug from the that of a transport regulator. The World Health Organization (WHO) organism from its hostile environment of viruses. sible for this barrier (Bouwstra 1997). It is painless compared to needles delivery systems for sensitive drug classes. This only aggravates the problem of the body. the use.5) and low MW (<500 Da). Prausnitz et al. efficacy. 2004) that use some form of physical . 1999. tion coefficient. 2000. Langer 1990). Despite the common flux of exogenous molecules into the body. safety. They are typically characterized by forefront (Drachman 1989. 2001. this challenge. A adults and children alike (Nir et al. estimates that 12 billion injections are given pathogens. Accidental needle sticks also add to the (15 μm) of skin. These are pharmacokinetics. Cropek and P. Skin routinely regu- blood plasma making repeated administration lates the flux of water molecules into and out of inevitable. the next ­millennia of providing sustained drug release for a prolonged health care will demand more accommodating period of time. The explore newer interfaces on the body for intro. skin is the first line of defense of an used method for administering therapeutics into organism and the last barrier separating the humans. eliminating first-pass hepatic metabolism and Denet et al. compliant. Delgado-Charro 9. Drug mol- system. makes use of human skin these high molecular weight hydrophilic drugs. Evolved to impede the annually (Kermode 2004).154 D. Sivamani advantageous mode of drug administration by et al.and protein-based drugs that will ­metabolism. A large fraction of drug drug or formulation excipients to modulate drug molecules lie outside these bounds. Further. 2008). Ogura et al. skin performs a complimentary role. b. Vanbrunt 1989. 2000. divided into two categories: (1) physical approaches including but not limited to iontopho- resis (Panchagnula et al. biggest challenge in transdermal drug delivery ducing therapeutics. stratum corneum (SC). 2003) and unique hierarchical structure of lipid-rich matrix makes drug administration stressful (Breau et al. Prausnitz 2004. 2004. These advances can be broadly Thomas and Finnin 2004). A more radical approach has been to become the key therapeutics in the future. However. Several advances to this effect have been ecules currently administered via the transdermal made in the last two to three decades and novel route fall within a narrow range of MW and lipo- drug delivery systems have been brought to the philicity. small molecules that are fairly lipophilic (parti- tiple administrations on a daily basis. sonophoresis (Mitragotri and Kost 2004.1 Transdermal Drug Delivery and Guy 2001). is respon- limitations of needle use in developed and devel. It also permits the influx of a variety of needle pain especially for patients requiring mul. and electroporation (Pliquett 1999. microneedles Transdermal drug delivery (TDD) offers an (McAllister et al. Prausnitz 1997. high log P (>1. with embedded keratinocytes in the upper strata 2001). Barry made in the past couple of decades to overcome 2001a. and toxics. and sustained delivery will become transdermal delivery systems to cash in on thera- the key features desirable of any drug delivery peutics that meet these requirements.1. and therefore offers superior patient compatibil- Injections comprise the next most commonly ity. today is to open the skin safely and reversibly to dermal drug delivery.M. One such approach. In addition oping countries alike (Kane et al. 2007). and mostly peptide. Needle phobia is a significant issue in the movement of foreign molecules across it. Pillai et al. hepatic metabolism cal. A large contribution to these novel thereby taking advantage of the natural selectiv- systems appeared as modifications to the active ity of skin membrane. As a result there has been a natural bias of noninvasive.

especially in rela- on the face value. almost all exclu. The dermis is highly on molecular weight have shown limited success. ately before cornification. Another four dozen are in the develop- mental phases. 20–30 layers of corneocytes embedded in a nating than expected. forming dead. than a quarter of a century since the introduction of the first patch. The dermis provides most of the bulk and ­laurocapram (Azone®). Williams and Barry 2004). on skin structure and constituents is followed by nificant proportion of the total revenues from a description of different model systems and pharmaceuticals (Barry 2001a. Differentiating daughter cells. thin layer of epidermis stacked upon a much philic in nature. After more methods employed in this area. and a net- used to deliver a low molecular weight drug. The landscape of transdermal deliv. It is maintained by (Okamoto et al.2 Structure of Skin of dozen. has drawn countless since a huge fraction is made up of generic repli. Methods.e. the most widely studied toughness of the skin.. It houses sweat transdermal delivery technologies but it is being glands. vascularized and permeable. delivery strategies in light of the knowledge Each of these methods has its individual benefits gained in the last five decades in this field of and limitations. SC. The number of original publications with “transdermal delivery” in the title has well approached 1000 with the numbers rising rapidly. receiving all of its nutrients and tations. Elias and Friend 1975). make up these 80 odd products. cas of similar drugs. b). A sound engineering matrix of lipid. sebaceous glands. Only 11 independent drugs Breathnach et al. Most efforts to push the envelope thicker substrate. iontophoresis) nantly of a fibrous collagen meshwork that is has successfully entered the market share of sparsely populated with cells. these numbers are misleading tion to its barrier function. research. 9. hair follicles. a small but sig. On the other hand. extruded from the cells immedi- approach coupled with fundamental understand. The market the “nuts and bolts” in transdermal drug delivery for patch-based therapeutics has since grown to research to a new scientist. consisting predomi- Only one physical method (i. 1988. tissue. blood vessels. methods. form of models. Lashmar et al. flattened corneo- the practical realization of it has been less fasci. phology and composition of keratinocytes. 1989). the dermis.2 Scope of Review These numbers form one metric to indicate that new scientists continue to be attracted to this The early 1990s of the last century brought the field. and (2) Equally important is adoption of the right plat- chemical approaches that employ chemical for. has failed to gain clinical acceptance in disposing of its waste products by diffusional transdermal delivery due to its skin irritation exchange with the dermis. the keratinocytes undergo terminal rooted in strong scientific acumen even though differentiation. form the ing of a complex biological tissue is required. help but ask. and Measurements in Transdermal Drug Delivery 155 energy to modulate the SC ultrastructure. The epidermis is devoid of chemical permeation enhancer with high expec. The concept of transdermal drug delivery is Ultimately. there is a change in the mor- research still important today?” (Barry 2001a. the keratino- ery opportunities seems grim and one cannot cytes move outward toward the surface of skin. and measurement tech- mulations to modulate skin transport barrier niques to evaluate new and traditional transdermal (Sinha and Kaur 2000. b).1. researchers since the early 1950s (Blank 1952.9  Models. cytes. A short introduction $4 billion per annum worldwide. Although somewhat satisfactory Elucidation of skin structure. Wong continuous cell division in the germinative basal et al. layer. This review aims to provide a synopsis of first transdermal patch to the market. 1989. work of capillaries supported by the connective lidocaine (235 Da). the number of marketed trans- dermal patches has not exceeded beyond a couple 9. 1973. “Is transdermal drug delivery During this process. The human skin is a sandwich of two layers: a sively below 500 Da and characteristically lipo. Corneocytes continually exfoliate from the .

And not seems to be elusive as newer observations con- surprisingly. since this superficial layer is where tinually require modulating the physical picture of the barrier property of skin resides. 1998. Blank the “brick and mortar” model of Michaels et al. with permeation occurring by Fickian diffu. Norlen 2001. The membrane is dis- folding” model of Norlen (Michaels et al. the interior of the chains inhibits formation of a els of Swartzendruber and Fenske. ordered or fluid with high permeability to small Swartzendruber et al. the et al. 1966. Elias et al. Norlén years of close scrutiny of the skin structure. and der. play a critical role in the barrier (Matoltsy to most substances than the dermis. the “domain on the interior of the fatty acyl chains. and frequently as experiments indicated that lipids. “molecular lipid lamellae” mod. Fenske et al. for all practical purposes. shown to mis. thin-section. epidermis from the underlying dermis by heat Proteins held within corneocyte lipid envelopes stripping followed by enzymatic removal of the thus form the bricks held by the mortar of a con- live epidermal layer. ~20–30 μm (Elias and Friend 1975. These include the classic “brick Occasionally.M. This mosaic” model of Forslind. These membranes are typically composed 9. unsaturation (Fettiplace and Haydon 1980).1 U  ltra-structure of Stratum of phosphoglycerides. 1977. the gradient in penetrant concentration across Madison et al. accommodate minor nuances. work of Scheuplein et al. Moreover. 1968. Michaels et al. indicates that their membranes do not provide a formidable barrier to water or water-soluble mol- ecules. 1987. especially polar much as five. While newer models are being proposed to rized the locus and origin of the molecular imper. 1987). Early solvent extraction that the SC is at least three. The bricks occupy. SC. hydrophilic solutes and water. These bilayers. This model treats the a large number of permeants. Sweeney and Downing 1970). 1975. Engström et al.2. by far. 1994. 1967. Kitson et al. These measurements indicated ume of this assembly. The permeability of the entire epidermis was found to freeze fracture studies of the SC established con- be indistinguishable from that of the SC alone. the entire skin is. (Breathnach et al. A comprehensive all encompassing model SC has received by far the most attention. and cho- Corneum lesterol where the lipid fatty acyl chains extend to 16 through 20 carbons with a varied degree of Several models have been proposed for the struc. Separating the ded in a continuous. homogeneous lipid matrix.. the “single gel phase” methyl branching coupled with unsaturation in model of Norlen. exist throughout the SC. the coarse macro- meability of skin and established it to be a passive scopic–microscopic structure is well agreed upon. Through their studies of the term itself coined by Elias (Elias and Friend the permeability of excised human skin in vitro to 1975. Cropek and P. 1975). rather than biologically active property The most simplistic model in this respect is still (Scheuplein 1965. Since the dominant resistance to Squier through freeze-­fracture. 1994. 1978. and “­ membrane highly ordered membrane. Bouwstra et al. 1989. methyl branching is also observed and mortar” model of Michaels. Menon et al. Scheuplein et al. they were able to skin barrier as a simplified two-compartment sys- show conclusively that the principal barrier to tem with discontinuous protein pockets embed- permeation is provided by the SC. Matoltsy 1986. measured tinuous lipid phase in the “brick and mortar” the permeability of the residual SC and dermis model. 1998). clusively that lipids form multiple broad bilayers This prompted Scheuplein to model the skin as a filling the corneocyte intercellular spaces three-layer laminate of SC. 1987. orders of magnitude less permeable lipids. 1972. Menon and Elias 1997. sphingomyelin. Odland 1983. Norlén et al. Interesting to note . In the last 60 1998. and Scheuplein 1973). Wertz et al. ture of the SC. The seminal this membrane (Wertz et al. local. the larger vol- independently. Karande SC to maintain a constant thickness of this layer at Forslind 1994. 1995. and permeation of most compounds is offered by the tracer studies (Squier 1973. provide the barrier to sion of the penetrating species through the three water permeability as determined by Elias and layers in series. 1973). the 2001). A general observation of mammalian cells ized within the SC.156 D. Downing 1992). epidermis. conclusively summa.

The molecular structure tion that barrier recovery was severely inhibited and composition of these constituents play an in skin treated with an enzyme that inhibits cuta- important role in defining the barrier properties neous cholesterol synthesis (Feingold et al. there is a considerable chain length 1939. 6 % of monounsaturated and 1 % of diunsaturated rated with the exception of ceramide 1. Baden and variation in the ceramides.2. Draize 1942. Methods. 1990). Abraham et al. ester-linked fatty acids unusual in consisting predominantly of very long (ceramide 1) are also present in the epidermal chain (20–28 carbons) saturated acids. In addition.9  Models. 1958. The ceramides are straight and satu.2. for providing a low pH or acidic surface (Blank In addition. 5. In addition. in 1990 based on their observa- Hedberg et al. characteristically amphiphilic in nature capable of extensive hydrogen bonding. Long et al. unsaturation is placed exclusively at the polar Presence of fatty acids along with cholesterol and end of these ceramide chains thus providing little ceramides is essential to the barrier property of room for formation of kinks. 1985. 1985. 2001). free fatty acids are also responsible to the membrane formed from these ceramides. cholesterol broadens phase transition Ceramides (1–6) constitute ~42 % of the material regions or in some cases may entirely abolish in the bilayers followed by cholesterol (~40 % subtransitions between gel phases thereby stabi- along with cholesteryl sulfate and cholesteryl lizing them (McMullen and McElhaney 1995.1 Ceramides addition. .2 Cholesterol Protein pockets.e. The (ceramides 4. Cholesterol also increases the chain mobility of lipids in the gel state making them 9.2. Pathak 1967. α-hydroxy acids them are a result of hydrolysis of ceramides. i. amounting to 25 % of extractable nent of the SC. 2. Wertz et al. 1985). 4. showed that choles- Subsequent sections provide a brief descrip. the bricks in the “brick and mor- Free cholesterol is the second most abundant tar” model. Cholesterol plays a key role in providing barrier Instead. thereby making it a physical as well as physio- able during bilayer formation. This may be This provides room for interdigitation of the critical to the antimicrobial activity of the SC hydrocarbon chains. the amide-linked fatty acids include nonhy. Wertz 1986). 1993). form the second important compo- lipid in the SC.. from 15 to 48.2 Lipids of Stratum Corneum more pliable and thus. of the bilayers and in turn of the SC membrane. and Fatty acids make up ~13 % of the SC lipids. with only ceramides. 1987. esters) and fatty acids (~13  %) (Wertz and Takahashi et al. Also the acids (Wertz et al.2. Yardley 1983. potentially. Downing 1983a. 9.2. The 5) and phytosphingosines (ceramides 3 and 6). Later Takahashi et al. 1996). an interaction highly favor. terol at high concentrations (>30 % molar basis) tion of the corneocyte proteins and the lipids of promotes lamellar structures. and ceramides (Wertz et al. property of the SC. 1996). The ceramides 9. Summerly 1981. origin of these fatty acids is not completely Also. 15 % of the SC lipids are made acyl chain structures in the SC (Yardley and of cholesteryl sulfate and cholesteryl esters.2. 1974). Also. Downing 1992). the SC bilayers are made of cholesterol. In 9. once again a primo in formation of self-assembled lamellae. This architecture is the SC. and 6). 1987. 1988). to provide superior barrier properties (Takahashi et al. and Measurements in Transdermal Drug Delivery 157 is the lack of any phospholipids or the usual fatty lipid. the lipids are logical barrier (Fluhr et al. Qiang et al. These pockets are included in flat. In addition to providing structural integ- well poised to provide a highly ordered structure rity to the SC. by Feingold et al.3 Fatty Acids include both sphingosines (ceramides 1. Beare et al. more resistant to mechanical stresses (de Kruyff et al. b. understood. regarded generally the SC.3 Proteins of Stratum Corneum 9. and ω-hydroxy acids composition of the mixture of fatty acids is (ceramide 1). although it is believed that some of droxy acids (ceramides 2 and 3). This was shown conclusively fatty acids.2.2.

Shelley contain mainly 30–34 carbon ω-hydroxyacyl and Melton 1949. Early studies by many Green 1981.158 D. Earlier in epidermal SC. These include: (a) diffusion through append- differentiation. persists even after extensive extraction with Histochemical techniques. Keratins 9. Karande hexagonal. 1982. This helps the sphingosine moieties to experimental system. b). These hydroxyceramide molecules applied chemicals (Mackee et al.1 Skin Appendages inner portion of the envelope consists of cross linked proteins. The appendageal area available for diffusion by disulfide bridges that can be solubilized only is significantly lower. (b) diffusion through the corneo- polypeptides are found in the SC (Skerrow and cytes of the SC. They are the SC. The corneocyte envelope enclosing the keratin filaments is made of two layers. The SC proteins are typically composed of covalently bound hydroxyceramides can inter- keratin. and predominate. 1973). The keratin in the SC is prob. Kao et al. 1988).3. whereas higher molecular weight hair follicles. Tregear chains and represent 2 % of the dry weight of 1961. As a result. when all the intercellular lipids are extracted the cytes. the detailed and lipids of the intact SC comprises the predom- structures of which are still under investigation inant route through which most molecules pene- (Steinert and Cantieri 1983). relatively poor in cysteine. considerable attention as an important perme- ably responsible for maintaining the hexagonal ation pathway for ions or large polar molecules shapes of the corneocytes and may contribute to that have slow permeation across the SC (Barry the toughness and flexibility of the SC (Wertz and 2001a.M. Vankooten and Mali 1966. Using full-thickness mouse skin main- protein envelope through the ω-hydroxy termi. The lipid been used to visualize and quantitate appenda- envelope hydroxyceramides anchor the corneo. 1945. Simon and Green 1984). Downing 1989). Wertz and Downing 1987). Wertz and Downing 1989). The 9. Individual keratin molecules ­lipids of the SC. tained as short-term organ cultures in an in vitro nus. rich in serine and gly- cine. geal absorption. predominantly involucrin and at The involvement of skin appendages in transcu- least six other soluble and membrane-associated taneous permeation has received considerable proteins (Rice and Green 1977. This may explain why unlike pyrene was higher in haired mice skin compared other membranous structures the lipid envelope to hairless mice skin (Kao et al. Cropek and P. low molecular weight keratins ages such as sweat ducts. Watt and ­ attention over six decades. At least Rutherford and Black 1969. Wahlberg 1968. These studies revealed that topi- cytes to the intercellular lipids. 1987. Diffusion across the corneocytes aggregate to form superhelices. demonstrated interdigitate with the lipid lamellae (Wertz and that permeation of topically applied benzo[a] Downing 1987). Fredriksson 1961. and physiologically dead corneo.1 %. They are stabilized trate. sebaceous glands. Keratins are a family of α-helical poly. digitate in a zip-like manner to close the inter- peptides ranging from 40 to 70 kDa in size (Green cellular space and thus maintain the integrity of et al. Wallace and Barnett 50 % of the hydroxyceramides are linked to the 1978). autoradiographic methanol–chloroform mixture (Swartzendruber techniques. and fluorescence microscopy have et al. the SC (Wertz and Downing 1989). The outer investigators implicated skin appendages as portion is made of ester-linked ω-hydroxyacyl­ important avenues for penetration of topically sphingosines. and (c) diffusion through the Hunter 1978). but has received by reducing agents. and contain N-acetylserine at the amino ter- minus (Steinert and Cantieri 1983). ~0. even cally applied agents concentrated and persisted in .3 Routes of Permeation accumulate throughout epidermal differentiation and represent the major component of the SC as There are three major routes of permeation for well as of epidermal appendages such as hair and passive diffusion of a molecule across the nails (Baden et al.

2005). Only ids (Barry 1988. the secretory coils of the 9. 2006). and Tenjarla et al.3 Intercellular Route sweat glands themselves lack cornified layers but are rich in tight junctions as evidenced by Several chemicals can alter or disrupt the organi- the colocalization of occludin and claudin-4 zation of lipid molecules in the SC bilayers (Hashimot 1971a. Hexamethyl sulfoxide and dimethyl sulf- Hydrocortisone permeability increased in tissue oxide convert α-helical keratins in the corneocyte engineered skin on insertion of hair follicles to β-sheets (Oertel 1977). 2004). Dithiothreitol enhances flux of sucrose and man- ally and morphologically distinct (Wilke et al. have demon- (Michel et al. Certain permeation enhancers can open up the Of all the appendageal routes.3. 1999). porous pathways for diffusion across them. Lee et al. and Measurements in Transdermal Drug Delivery 159 the hair follicles and sebaceous glands (Grasso 9. which seem to be absent from the epi- dermal duct lining (Hashimot 1971a. The innermost layer of the intra. 1992). increasing transcutaneous permeation by affect- cein (Lieb et al. (Hueber et al. (Vankoote and Mali 1966). In con. It also serves as an example. 1984). and melanin (Li and ing secondary structure of keratins within the Hoffman 1997) to hair follicles. Lee et al. In light of these observa. 1994). 1992). neocytes for polar substances after pretreatment nified corneocytes but contain luminal tight of the skin (Eder and Müller-Goymann 1995). 2001). nitol across the SC exclusively through interac- 2005. dense keratin structure in corneocytes creating cle has received the most attention as a promi. cal. proposed acid enhances the permeability of verapamil by that the active permeation barriers in sweat interacting with skin proteins (Shah et al. ducts in the epidermis and dermis are function. junctions. Tenjarla et al. Of these.2 Intracellular Route and Lansdown 1972. tory creams in disrupting intracellular keratin tigated with great success. Barry postulated different ways in ducts and the sweat glands could serve as poten. 1991. ton microscopy (Lee et al. Fluidity caused at the polar plane due to the . 1994. Wilke et al. 2008). 2006). Following up with elegant β-sheet confirmation (Xueqin et al. 2006). 2008). b). Enhancers can act a few experimental studies have actually been on polar head groups of lipids and modify the dedicated to evaluating the contribution of sweat hydrogen bonding and ionic forces between them glands and ducts to transcutaneous permeation resulting in a disruption of the packing geometry. Lauric immunostaining studies. Similar to the dermal ducts. b). For nent route of permeation. He et al. 1965). the hair folli.9  Models. showed that the corneocyte (Cooper 1982). have 1993). the dermal ducts lack the presence of cor. Wilke et al. Dimethyl sulfoxide penetration of corticosteroids is considerably can induce reversible changes in protein struc- lower in hairless skin compared to haired skin tures of isolated corneocytes (Mendelsohn et al. which permeation enhancers can modify SC lip- tial permeation routes (Wilke et al. decylmethyl sulfoxide interacts with important cutaneous reservoir for topically keratin and is hypothesized to enhance permea- applied molecules (Lademann et al. liposomes matrix and the protein envelope using multipho- have been shown to deliver DNA (Li et al. 1979. 1999). Holland et al. 2006). plasmids (Domashenko et al. Azone® can act on reported that the auxiliary SC associated with the keratin fibers of the corneocytes converting the sweat glands has a reduced barrier function their rigid α-helical conformation to a flexible (Lee et al. Methods. Foreman et al.3. 2000). bility by opening up aqueous channels within the Hueber et al. Hashimot et al. propose that dermal sweat across the SC. Permeation enhancers that strated the capability of thioglycolates in depila- specifically target hair follicles have been inves. myristate increase the permeability of the cor- trast. shown that N-trimethyl chitosan is capable of monoclonal antibodies (Balsari et al. tions with corneocyte keratin matrix (Goates and epidermal duct is completely keratinized Knutson 1993). corneocytes (He et al. Oleic acid and isopropyl (Zelickson 1961. thereby facilitating the diffusion of molecules tions.

In addition to fluidiz. Enhancers can also insert dermis and dermis offer minimal resistance to themselves between the hydrocarbon chains of passive diffusion of solutes. excellent sources of human skin (Bronaugh et al. 2006. Interestingly. Loss of the comparison between in vivo and ex vivo SC lipids from the SC was monitored as a decrease in and have shown that it retains its barrier proper- the signal intensity of methylene groups of lipid ties for several days after harvesting (Berenson chains in Fourier transform infrared (FT-IR) spec. or dis- ation of aqueous pockets that facilitate diffusion carded skin from breast reduction procedures are of hydrophilic molecules. Wester and Maibach 1989. 1984). fatty amines. expected for ex vivo skin to maintain the barrier ants. (Harrison et al.M. Human skin is the obvious choice in experiments Robert Peter Chilcott 2000). 1999. fatty −18 °C for 466 days did not show any significant esters. Several reports have documented from bilayers or the corneocyte envelope. zwitterionic surfac. intuitively lipid bilayers facilitates the diffusion of perme. Consequent fluidization of the tinized corneocytes. bilayers. Barry et al. Chilcott et al. cadaver skin. as lipid ­extractors or lipid fluid. fatty alcohols. and Burch 1951. Some investigators have indeed veri- extent. safety concerns. Cropek and P. Barry tants. Norlen et al. The metabolic activity signal intensity of methylene groups (from hydro. human skin in permeation experiments there are zation correlated very well with the extent of skin several problems associated with its use such as permeabilization. fied. studied permeation enhancers without significant deterioration of barrier prop- from eight different categories: anionic surfac. Galey et al. across the skin is the SC. fatty acids. lim- ited supply. therefore. and regulatory considerations. 1976). Wester et al. An alternate consequence of disrupting 2006. showed a decrease by day 2 but stayed steady carbon tails of the enhancer) and disappearance of until day 8 (Wester et al. more simply. Disruption in packing of lipid chains can in integrity of the SC for an extended period of time turn alter the packing of polar head groups of the after harvesting when stored under appropriate lipid molecules. or micelle forma. ing bilayers. The SC is composed the lipid bilayers and thereby disrupt the packing of lipids and terminally differentiated.160 D. et al. Lipid fluidizers increased SC permea. cationic surfactants. Also the permeability measurements obtained on 9. fully kera- of lipid molecules. 1998a). Harrison et al. to a small conditions.4 In Vitro Skin Models human skin samples vary greatly between indi- viduals as well as between samples from differ- 9. diffusion of solute molecules across the lipid McCullough et al. activity was highest during the first 18 h after the Fluidization was monitored as an increase in the skin was harvested. Karande d­ isruption of packing geometry accelerates the c­ ompounds or therapeutics (Rao and Misra 1994. group interactions can cause extraction of lipid The primary barrier to transport of molecules molecules. and azone-like molecules. found that human cadaver skin stored at tants. It is. erties (Franz 1975. thereby accelerating. Friend 1992). monitored glucose metabolism in skin as a mea- bility by partitioning themselves in the bilayers sure of its viability and showed that the metabolic and disrupting the bilayers packing structure. Lipid extractors 5000 years old and buried in glacial ice was very increased SC permeability by extracting lipids well preserved. excised skin from autopsies. the diffusion of permeants. fied that skin can be frozen for up to 12 months Karande et al. have for determining the permeability of model shown that there is a statistically significant .4. 1984). Extent of extraction or fluidi. Kim et al. troscopy. also found that the skin obtained from an iceman izers (Karande et al. 2008).1 Excised Human skin ent anatomical sites on the same individual (Wester and Maibach 1992. In comparison the epi- tion (Barry 2004). 2005). Elewski 2007. Suppasrivasuseth et al. and showed that change in permeability toward tritiated water chemicals in all these categories could be classi. difficulty in procurement. enhancers that disrupt lipid head 1986. peaks related to the ordered packing of lipids in In spite of the several advantages of using FT-IR spectroscopy. phase separation. Freshly packing geometry of lipid head groups is the cre.

epidermis thick- by other investigators (Southwell et al. and butyl para. Tipre and skin exposed to water increases only twofold in Vavia 2003. porcine (Panchagnula effect on human skin but can increase hairless et al. Bronaugh was higher than that observed within the same et al. Roy between sites on the same animal (Elias et al. 1997). 1983). imental purpose (Friend 1992). Dick and Scott 1992. 1997. Zhao et al. porcine et al. have documented subject. Skin from experimental animals is different from ben on skin samples from several donors and human skin in thickness. 1989. These parameters vary significantly non-Gaussian distribution (Williams et al. Panchagnula et al. of permeation enhancers on skin permeability 2002). Hai et al. and con- found interdonor variabilities between 33 % and stitution of the SC. 2008). between the different species studied. 1997). Ogiso et al. 1992. Barry 1988a. underestimates the effect 1998).2 Excised Animal Skin both compounds have similar permeabilities across human skin. Karande et al. surement was obtained. 1997). rabbit 10 days. methyl paraben. Several Chilcott 2000). hairless mouse skin (Bond and Barry 1988a. sheep permeability (Panchagnula et al. dog overestimates the effect of permeation enhancers (Sato et al. 2004. Panchagnula et al. (Panchagnula et al. Panchagnula et al. c). is closest studied to represent human skin. Hairless mouse skin model and Degroot 1994. 2004. 2001. 1997. SC thickness. 1997). chirality. 1997. Animal skin to human skin in terms of its biochemical compo- offers advantages over human skin in that the age sition and histological features (Gray and Yardley and sex of the animal can be controlled as well as 1975. the inter. and to some extent the One needs to be cautious. Akomeah measured the permea. in extrapo- dietary habits of the individual (Robert Peter lating animal skin data to human skin. hamster (Coutelegros et al. 1991. 1997. and distribution and density 44 % (Akomeah et al. cow (Panchagnula 1990). mouse skin permeability by 15-fold (Bond and 2007). 1997. 1984. 1986. The lipid con- skin. Netzlaff et al. 1981). Rochefort et al. and particularly porcine ear skin.9  Models. c). Panchagnula et al. Artusi et al. Cho et al. 2008. animal skin is routinely used as a model for tent of the skin is a major determinant in its bar- human skin in in vitro experiments (Haigh and rier potential and differs between species or Smith 1994). Similar observations have been reported follicular density. Methods. their permeabilities across In view of the difficulties associated with human other skin models vary drastically. Further. In general. 1998b. however.4. 1997. et al. 2006b). While permeability of human guinea pig (Panchagnula et al. frog (Dewhurst mal skin permeability is higher than human skin and Williams 1993. 1995. water and 7-hydroxycoumarin. ness. 1994. fish (Watanabe et al. 1997). on skin permeability by sevenfold (Bond and Rohatagi et al. Hairless mouse skin which is com- et al. Smith 1993). While 9. 2008. A 2-min treatment with acetone has negligible Elgorashi et al. 1986). For two Cornwell and Barry 1995). another common 1992. 2008). Suh and Jun when compared to human skin (Rigg and Barry 1996. composition. and Measurements in Transdermal Drug Delivery 161 v­ ariation in the skin barrier property with relation large quantities of skin can be obtained for exper- to gender. Roy Barry 1988a. In contrast. Ben-Shabat et al. 2007). b. it has been observed that ani- et al. Masson et al. Bach and Lippold model. and marmoset (Scott Of all animal skin models studied. differences exist and have been documented. Panchagnula et al. Wester et al. bility of caffeine. d). rat (Panchagnula monly used as a model for human skin is com- et al. of appendages such as sweat glands and hair fol- subject skin sample variability in skin permeation licles (Schalla and Schaefer 1982. . In general. 1997. shed snake skin. 2005. and full skin thickness for 16 animal mod- Langguth et al. 2008). 1993. Pabla and Zia 2007). els including human skin (Panchagnula et al. model compounds used in this study. snake (Megrab et al. b. 1997. paratively fragile. Panchagnula et al. hydration can completely disintegrate (Panchagnula et al. lag time and permeability varied significantly across the skin models. these permeability measurements show a 1997). monkey (Wester and Maibach 1987. b. 1991) are some of the animal skin models skin. Sebastiani et al. c. Mouse (Bonina et al. Bhandari 1980. time of the day when mea.

of drugs. 2002. or combinations thereof (Slivka et al. 1998. and SkinEthic® models or split thickness skin by enzymatic digestion based on their morphology. 2008) by MatTek Corp. Ashland. 2008).e. EpiSkin® and SkinEthic RHE® (Botham The biggest advantage of cell culture models. Hughes et al. using trypsin (Larsen et al. there is also a variation observed in skin achieves terminal differentiation and manifests permeability with age and anatomical location characteristics remarkably similar to those of nor- within the same species (Bronaugh et al. 2006).M. corrosivity. Keratinocytes receive nutrients from of the same permeation enhancer on account of the lower surface of the culture while being the differences in their constituents. and cost savings when compared to animal use. well-defined end points. i. follicular structure and density lagen lattices (Bell et al. In general. 1995. 2004). irri- Basal keratinocytes are isolated and grown at an tancy. zation studies (Audus et al. Sobral show varying permeability responses in presence et al. is their amenability to high-throughput et al. Cropek and P. 2002). 2004. Porcine skin resembles human skin thetic membranes such as nylon mesh (Slivka most in terms of the SC thickness (Holbrook and et al. 2007. Jacobi (Fransson et al. dead de-­epidermized der- mis (Regnier et al. Organogenesis. 2004. 2002. 1990). Jacobi membranes (MonteiroRiviere et al. 1996. 2008). 1979). 2004. Labs. Kandarova et al. glycated collagen (Jacobi et al. Elyan et al. 2005. Today. 2007). Shibayama et al. tests for evaluating phototoxicity. Jacobi been used successfully include permeable syn- et al. 1989). lipid composition (Gray and (Pageon and Asselineau 2005). 1997. EpiDerm™ and rapid assessment of permeability and metabolism EpiDermFT™ (Hayden et al. 2007) by SkinEthic studies for drug discovery or formulation optimi. col- et al. and transport properties (Netzaff et al. Crooke et al. 2007). 2008. France. (Simon and Maibach 2000). Lotte et al. EpiSkin®. biodegradable scaffolds (El Ghalbzouri et al. Poumay et al. Netzlaff et al. 2006). collagen-­ Yardley 1975). The architecture. Mansbridge 9. 2005). 1998. or thermolysin (Walzer et al. epidermis thickness (Wester and et al. fibrin sentative model for human skin. and the underlying vasculature glycosaminoglycan matrices (Boyce et al. have reviewed the with obtaining keratinocytes from full thickness EpiDerm™. completely cornified cells sur- Dupuis et al. representation (Roguet et al. composition. 2000. 2004). MA. 1988). 2003.. 1982. 1981). air–liquid interface on a substrate that is equiva. 2007). stricter control over experimental condi. 1994. Schafer-Korting et al. Cell culture or tissue culture-­based Sekkat 2001). Faller and These include TestSkin® and TestSkin® II by Bracher 2002. Sandby-Moller et al. Karande Sekkat et al. several advantages such as high reproducibility. Luu-The however. Netzlaff et al. Moody in vitro cell culture models of living tissues offer et al. 2008. As a result. Morikawa et al. the por. 1993. Kyoto. Schafer- tions. 1998. Duncan rounded by a lipid intercellular matrix (Nabila et al.3 Living Skin Equivalents 2002).162 D. 2007). collagen Maibach 1989. Wester and Maibach 1989. the topmost layer variation. In addition to an interspecies entiation. 2006. homeostasis. Nice. mal SC. 1988). 2002). Skin samples obtained from different species 1993. 2007) by Gunze. lipid composition. pushed upward in a process of progressive differ- and microstructure. 1986. Canton. syn- thetic scaffolds (Shakespeare 2001. and potential time Korting et al. matrices (Shahabeddin et al.4. 1996) and polycarbonate Odland 1974. Muhammad et al. Vitrolife-Skin (Uchino et al. chitosan cross-linked collagen-­glycosaminoglycan cine skin has gained wide acceptance as a repre. In 14–21 days. Barker et al.. 1990). 2007. Kandarova et al. dispase (Green biochemical markers. Rodriguez et al. MA (Davis 1990. several cell culture-based models of human skin can potentially overcome skin models are commercially available for ready this problem by offering a more consistent skin use in skin permeation or skin toxicity studies. 2007). Lee et al. Rehder et al. and their applicability in et al. Borgia et al. Flamand et al. Reconstruction of skin in vitro typically starts Japan. 2004. 2004.. (Holland et al. and lipid lent of the dermis. Dermal equivalents that have composition of these models come close to human . 2004.

EpiSkin®. In et al. Skinethic™ HRE was chemically defined media and sources for cells approximately sevenfold more permeable com. EpiDermTM was the most resistant to SLS and EpiDerm™. Schmook et al. commercially available models use proprietary pared with human skin. Ponec et al. 2002). human and pig skin as (Vicanova et al. report that full-thickness models based on cutaneous bioavailability studies on topical for. b). Furthermore. cited as reasons for this poor performance. Beastall et al. 1980. 1986. Impaired desquamation (Vicanova icylic acid. In addition. found that the EpiSkin® cal factors such as hydrogen bonding. The biggest shortcoming of commercially ­compared the permeabilities of four topical der. Although the reconstructed human skin 9. across rat. and caffeine follow the same rank such as molecule size and shape. but clotrima. Epiderm™. impaired transfer of desmosomes terbinafine. a multilab study verified treatment with sodium lauryl sulfate (SLS).9  Models. mannitol. mostly to industry and out of reach of most aca- zole flux across it was 1000-fold higher and its demic labs and small enterprises. all skin concentration 50-fold higher when com. available skin models is their relatively weak bar- matological compounds of varying polarity—sal. that the permeability ranking across EpiSkin®. ity of using such models. In these studies pig skin performed similar Another significant impediment to the use of to human skin with comparable flux of solute reconstructed human skin models is their high across both tissues. Methods. Dreher et al. vastly different in EpiDerm™ and EpiSkin® mod- els compared to ex vivo human skin (Dreher et al. have shown that and Smith 1994). Faller et al. hydrocortisone. 2008). b). 2001). vehicle effects were observed to be situation more closely (Nakamura et al. a tamic oxaloacetic transaminase (GOT) and lactate liposomal dispersion and a hydrogel (Dreher dehydrogenase (LDH). 1996a. compared the models in and α-tocopherol acetate from a water in oil their ability to secrete extracellular enzymes glu. clotrimazole and et al. 1997). Lotte et al.4. 2002).4 Polymers models underperform significantly in reproduc- ing the barrier properties of ex vivo human skin Model membrane systems can provide tremen- they can still be used to rank order the permeabil. 2002). an oil in water (o/w)-emulsion. dous insight into mechanistic details of solute dif- ities of solutes based on their permeabilities. Nakamura thickness human skin (Marty et al. 1996a. organ cultures of skin explants match the in vivo mulations. 2002). 1999). and Measurements in Transdermal Drug Delivery 163 skin (Ponec and Kempenaar 1995. and presence of unke- well as two models of human skin—Graftskin™ ratinized microscopic foci (Mak et al. and chemi- et al. the reconstructed skin models have epidermis for caffeine and testosterone (Schafer- higher permeabilities compared to excised human Korting et al. order permeability as human skin for caffeine 2000. Diffusion of a solute molecule the skin absorption and permeability of lauric across a membrane is governed by physical factors acid. 1991) are LSE™ and Skinethic™ HRE (Schmook et al. comparable to the permeation through human In general. and SkinEthic™ RHE study Marty et al. rier function. Flynn 1985. and electrostatic interactions. skin (Gysler et al. reported that trinitroglycerol are based on the epidermis raised on a minimal and estradiol were about 20-fold more permeable dermal equivalent such as collagen gel scaffold across Skinethic™ HRE compared to split-­ encapsulating fibroblasts. pore order as they would on ex vivo human skin (Lotte distribution. (w/o)-emulsion. In a similar els. and Skin Ethic™ RHE models was most reproducible (Faller and Bracher 2002). and interleukin-1α on et al. In contrast. Graftskin™ LSE™ provided cost. The . pore size. All three leading mod- fold more permeable to clotrimazole. Haigh different molecules. fusion and thermodynamics of solute–membrane Such a rank order has been shown to match the and solvent–membrane interactions (Corrigan order obtained on ex vivo human skin for several et al. 1990). This has limited the use of such models an adequate barrier to salicylic acid. hydropho- and EpiDerm™ models showed the same rank bic interactions. path length and tortuosity. that can put additional constraints on the flexibil- pared to human skin for salicylic acid and 900.

have used polyacrylamide gels to porous substrate. was 1000-fold lower Cross et al. ethyl vinyl acetate (Farinha et al. Miyajima et al. 1984. polyvinyl chloride. 1978. and some other permeants across intact SC (Abraham ease of procurement. have used the SC barrier based on the permeability of a filter supports or filter supports filled with organic model hydrophobic drug. polyether sulfone membrane (Hadgraft and Ridout 1988). Reasonable correlation was have been proposed to quantify skin permeability. Drug permea- In spirit of the “fluid—mosaic model” of the skin. Dyer et al. Guy 1988). Transport resistance across the polymers such as silicone (Hou and Flynn 1997. 1992.5 Lipids In vitro models based on lipids. pal- and Bronaugh 1983. 2002. multimembrane laminates (Scheuplein liposomes composed of SC lipids: ceramides. Although such have reported studies on membranes reconstituted models are simplistic and lack all the functional from porcine SC lipids or porcine brain ceramides and structural complexity of skin. 1979). however. Kuempel et al. 2006) have been used to this end. Viegas et al. developed a model skin membrane by fixing et al. 2001). ether. Relatively less studied synthetic mem. 2003. Houk and Guy 1988). 9. 2003). esters. linoleic acid. model membrane. et al. and paraffins have also been although permeability through the model system used to model diffusion through skin (Houk and was an order of magnitude higher. Cropek and P. 2006). Biodyne B (Matsuzaki (Ottaviani et al. stitute (SCS) by applying a mixture of synthetic Turakka et al. Schramm. Matsuzaki (Farinha et al. 2003). 1987).88) with that through guinea pig skin chloroform. 2007). and a mitic acid. and tetradec- Du Plessis et al. they provide on porous substrates. These models have been several other advantages such as uniformity of shown to reproduce the permeability of water and structure. 1994). phosphatidylcholine. A few studies. and cholesterol on a Baxter et al. and cholesterol-3-sulfate mixture of isopropyl myristate and silicone oil onto a supporting filter. The composition. 1996). 1998. however. Other groups model systems (Dyer et al. filter sup. constructed a model lipid matrix from cholesterol. de Jager et al. poly(dimethylsiloxane) (Cronin et al. b). In diffusion water. 1998). A three-component mixture of dipalmitoyl 2003). 5-fluorouracil. Friberg et al. Using a liquid to study diffusion of topical agents (Tanaka similar approach. and Downing 1989. Moghimi et al. model human skin and study the energetics of liq. or 9.164 D. 2004). and free fatty acids of the SC and their studies across model biomembranes. This model ports have typically gained prominence as support matrix was shown to be a good representation of membranes. Synthetic membranes and excised skin. Farinha et al. Friberg and Kayali 1989.M. Karande contribution from each of these factors can obtained between diffusion of a wide range of ­potentially be decoupled by a systematic study compounds across the IPM membrane and with model membranes. created a SC sub- et al.4. cholesterol. Several methods epidermal barrier. sodium salts (Moghimi et al. utmost importance for percutaneous absorption and ing diffusion cell has been used to simulate the transdermal delivery applications. Farinha et al. Kittayanond et al. SC lipids. and orientation of lipids in the SCS bore high uid jet penetration into skin (Schramm-Baxter resemblance to that of the intercellular barrier lip- et al. sample-to-sample reproducibility. Demeere and Tomlinson 1984.5  valuation of Skin E mixtures of natural or model lipids have been Permeability In Vitro evaluated for studying percutaneous absorption in humans. bility through this system correlated very well organic solvents such as 1-octanol. 1993. have tested zeolites as ids in SC (de Jager et al. free fatty acids. . Lieckfeldt et al. Barry and Brace 1977. Frum ane showed an order of magnitude improvement et al. model lipids. organization. 1990. 2006a. polyvinylidene difluoride (Olivella in transport resistance when compared to IPM et al. cellulose acetate (Barry and as compared to excised skin (Hadgraft and Ridout Eleini 1976. brane systems are porous materials. alkanes. An artificial lipid membrane composed The ability to measure skin permeability is of of isopropyl myristate (IPM) supported in a rotat. 1986). 1993. (r = 0.

L is the path length of diffusion.9  Models. Mixing . chromatography. (9.5.. A typical diffu- sion cell assembly contains a donor chamber L2 tL = coupled to a receiver chamber by means of a exhibits a lag time.4) C0 receiver chamber is then expressed as a permea.1) above can A wide variety of diffusion cell systems have been be rewritten in a simpler form as developed for measuring solute permeation through membranes (Frantz 1990. meability resulting in misleading conclusions.5.5. The membrane. and ute diffusing across skin. needs to be detected in the receiver chamber by means of appropriate ana- é Dt 1 2 ¥ æ .2) L ë 6D û The most common configurations are vertical cells. is The terms K.1 Diffusion Measurements dM t C0 KD = (9. or chemical properties. ú (9. in this case skin.Dn 2p 2 t ö ù M t = C0 KL ê 2 . Care needs to be taken that where C0 is the concentration of the solute in the labeling of the solute does not alter its physico- donor chamber. across the skin.4) A number of relationships have been used to above provides a straightforward way of deter- describe the permeation of drugs across skin.5. and Measurements in Transdermal Drug Delivery 165 9. Eq. mining skin permeability to different solutes by While the basis for these relationships can be measuring their flux. etc. P = . Nevertheless. Mt = ê t . Friend 1992). which may affect skin per- log P of the solute into skin (SC). the skin donor and the epidermis/dermis to the receiver.1) as ELISA. A L solute whose permeability across skin is to be permeability. receiver chamber are arranged side-­by-­side.1. The solute. These may include spectrome- ëL 6 p n =1 è L2 øû try. Solute permeability can then be measured is placed in the donor chamber by for.1. The rate of appearance of the solute in the P = dt (9. Bronaugh and Collier C0 KD é L2 ù 1991. D is the effec. ber separated by the membrane in between. D and L are grouped together sandwiched between the donor and receiver KD chambers such that the SC is exposed to the and defined as a single term. where the donor chamber is atop the receiver cham- This represents the permeability profile of a sol. Osborne 1986). estimated from flux of the solute across the skin mulating it in a suitable solvent. time. K is the partition coefficient. complex. (9. the amount of solute (Mt) crossing the skin in time t can be related to skin permeability 9. bility profile in the form amount vs. western blots. The permeability profile is linear in time but ation of solutes across the skin. whose permeability is to be assessed For an infinite dose of solute in the donor. The solute itself may be labeled using a fluoropore or radioisotope for direct detection. 9. Eq.2 å exp ç ÷ú lytical methods.1. Several simplifying assumptions have been made in deriving this relationship (Foreman and Kelly 9. 6D spring clamp or screw. The slope of this profile horizontal diffusion cells where the donor and provides flux of the solute across skin. Methods.3) dt L Diffusion cells are by far the oldest and most commonly used apparatus in measuring perme. tive diffusion coefficient across the skin. Appearance of and its concentration in the donor chamber. Gummer and Maibach 1991. whose permeability is to be assessed.3 Diffusion Cells At steady state ( t ® ¥ ) .2 Model Solutes (P) by a reasonably straightforward relation.1 Theory 1976. biochemical methods such (9. the solute in the receiver chamber is periodically dM t monitored using appropriate analytical methods.

Kelder 2003). 1985a. mixing is not critical. (b) Vertical cells sion into the subdermal vasculature) have also been Vertical diffusion cells are closer to used successfully (Ainsworth 1960). In case (Dyer et al. bovine serum albumin (Brown and receiver using a clamp. Whitton et al. However. 1986). Dugard et al. Chamber (Holland et al. Kao reported that long-term hydration in rodent skin. and flow when assessing the permeability of a hydrophobic through system with central inlet and periph- solute in an aqueous receiver compartment (Tsuruta eral effluent ports (Astley and Levine 1976). Tojo et al. 2007). metabolism and diffu. In 1977. ation of highly hydrophobic compounds.166 D. Tas et al. For studying perme. Ito et al. 2006. 1981. glass diffusion cells with high-solute concentration in the receiver chamber.M. vertical diffusion cells (Coldman et al. 1969). receptor fluid is continuously refreshed to mimic Soni et al. (Washitake et al. Karande of the chambers to create homogeneous compart. Loftsson 1982. 1989). 1984. 1984). National Laboratory Skin Permeability ation rates in smaller donor chambers.e. ical flasks configuration (Wurster et al. glass con- of vertical diffusion cells. L-shape configuration ments is critical in horizontal diffusion cells. equi-­compartment diffu- chamber better mimics in vivo conditions and pre. Valia– Formation of unmixed zones is especially critical Chien cells (Tojo et al. and Ulsamer 1975). 1986). The Franz diffusion cell cised when using solubilizing agents that they do remains the most widely studied vertical dif- not alter inherent membrane properties. Poloxamer 188 (Hoelgaard and studied a similar cell with the sampling arm Mollgaard 1982). solubiliz. 1985a). The water jackets or simply submerging the entire cell Coldman cell represents the earliest of all assembly into a water bath. Gummer et al. design had poor mixing properties which ered when assessing solute permeability over have been addressed in subsequent modifica- extended periods of time. 1980). 1987. Some studies have tions (Nacht et al. b. Skin is sandwiched between the donor and Stewart 1984). 1979). 1996). Bakand et al. Finally. In the past two 1973. 1979. These chamber has been shown to have some effects on include the release cells (Morell et al. Bond and Barry 1988a. 1996). Aramaki et al. systems discussed above. b). c. (a) Horizontal diffusion cells (c) Skin Flaps Several designs have been suggested and In addition to the conventional diffusion used successfully for this type of diffusion cell. 1983. The original effect on membrane integrity needs to be consid. several modified versions of these mixing can be obtained using small magnetic stir early designs have been used (Morell et al. and located at the bottom of the receiver (Whitton ethanol (Scott et al. Water penetration into the skin introduces a cells in combination with the Automatic lateral strain at the edges of the donor chamber due Sample Preparation with Extraction Columns to swelling. Bronaugh and Stewart 1984. and Ussing type chambers (Li et al. changes in permeation rates (Whitton and Everall Tiemessen et al. 2007). Efficient recent years. the area of diffusion of the donor cell have been used successfully. Hinz et al. can lead to Reifenrath 1986. and Everall 1973). PEG 400 (Valia et al. a homogeneous well-mixed receiver vertical membrane. Cropek and P. d. Hydration fusion cell today (Franz 1978). 2003. 2006. bars. decades several modifications of the Coldman 1989). These include Triton X-lOO (Bronaugh ber. This scales as the strain edge available system (Bosman et al. the permeation rates (Karande and Mitragotri enhancer cells (Bosman et al. in vivo sink conditions (i.. A glass cell with a side arm for sampling and ing solvents can be added to the aqueous receiver stir bar for mixing forms the receiver cham- chamber. Temperature in vivo situation in obtaining permeability control of the diffusion cell can be attained using data across the skin (Friend 1992). steel mesh (Southwell and Barry 1983). Flow-through diffusion cells in which the 1996. 1996). et al. novel in vitro sys- These include the T-shape configuration tems that measure effect of perfusion rates on . Hawkins and and in particular hairless mouse skin. Caution needs to be exer. 1984). 2006. sion cell with high area to volume ratio (Flynn vents the formation of a static boundary layer of and Smith 1971). Oak Ridge per unit area and results in higher observed perme.

8) 1963. ¶t ¶x where D is the average solute diffusion coeffi- cient in the SC. Drugs like corticosteroids 4 Pt were shown to localize within the SC (Vickers Cs ( t ) = C0 (9. 1998). 1991). 1990. Eq. where x = 0 corresponds to the SC surface and Isolated Perfused Porcine Skin Flap (IPPSF) x = L corresponds to the end of the SC. 1987. solute concentration can be mea- increasingly used in measuring drug concentra. sured via tape stripping in the SC to infer its tion and its concentration profile in the SC steady-state permeability. 1990. tography (HPLC) or an immunoassay or radioimmunoassay is required in conjunction to 9. an adhesive tape is applied to Cs ( t ) 4 DKt the skin and removed abruptly. ¥ useful in dermatopathological and dermatophar. Ohman and Vahlquist 1994).5. Bovine udder is used in a similar fashion for permeation studies Cs ( t ) ïì . For short times. These observa. Cs is given as follows (Crank 1975): Riviere et al.e. Eq. Tojo tional to its steady-state permeability. The observation that skin may serve as a reservoir for Using the definition of permeability P. Methods based on measuring solute diffusion ary conditions: across skin may not always provide the sensitiv- ity required to measure small perturbations in Cs ( x = 0 ) = KC0 skin permeability or follow permeability Cs ( x = L ) = 0 changes over short intervals of time. low values of L2 .D ( 2n + 1) p t ïü 2 2 ¥ (Kietzmann et al. and x is the distance from the SC surface. Equation (9. Typically. Pellett et al. 1997. 2001a. Electrical . SC. and Measurements in Transdermal Drug Delivery 167 solute permeation have also been designed. ¶Cs ¶ 2C = D 2s (9. Cs is the solute concentration in 9. i.2. and fused by the caudal superficial epigastric C0 is the donor concentration of the solute. An analytical tech- (Pershing et al.6) 9. This application » (9. This technique is now being Accordingly. 1983. and Lee 1989).8) shows that the solute concentra- nique in investigating the barrier and reservoir tion in the SC measured at short times is propor- function of the skin (Rougier et al.5. Solute diffusion in the SC can be described by Several studies have successfully applied this Fick’s law (Crank 1975) as follows method to determine the skin permeability of a wide range of solutes (Stinchcomb et al. average solute partition coefficient in the SC.2 Tape Stripping where C∞ is the solute concentration in SC at Tape stripping is a technique that has been found steady state ( C = KC0 ). 1999. The artery and its associated veins and mounted resulting equation for solute concentration in the on a diffusion cell (Williams et al.5. (9. 8 1 C¥ = 1- p2 å n = 0 ( 2n + 1) 2 exp í L2 ý îï þï (9. b. (9.1 Theory accurately determine solute concentration.6) can be simplified as 1999).7) can be repeated between 10 and 100 times (Sheth C0 L2 et al. 1993).5) Alberti et al.1) can be solved with the following bound. 2 macological research for selectively or at times Dt exhaustively removing the SC (Surber et al.9  Models.7) further simplifies to and Ferguson 1955). c. (9. Moser et al. the above chemicals was first reported in 1955 (Malkinson Eq.3 Impedance Spectroscopy the SC. L tions led to the use of the tape stripping tech. 2001). Carr and Wieland 1966).. K is the is a model system of porcine skin flap per. Shah nique such as high-performance liquid chroma- et al. Methods.

and skin permeability. is capable of assessing thousands of for- Z = ê R2 + ú (9. 1999. chemical enhancers on skin permeability Electrical properties of SC parallel those of per. 2003). electrical 1966). of a solute needs to be assessed by conventional 1998. 2006). As the skin is properties. Kumar and Lin 2008). have described the circuit containing a resistance R shunted by design of an impedance based high-throughput capacitance C (Lackermeier et al. which may be tedious. and more sensitive. A simplified correlation between skin resistiv- ity. 1999). Skin et al. which were capable of deliver- current (AC) signal. time consuming. The assay to determine the effect of chemical perme- impedance. (Karande et al. Tezel et al. 1960.5. b).1 Theory analysis. Karande et al. Tregear solute permeability in diffusion cells. In comparison to diffusion measurements of trol of current flow (Lawler et al. A review of factors governing the pas.4 Infrared Spectroscopy log P = log C . meability and play a dominant role in the con. One downside and the Nernst–Einstein relations for ideal solu.log R (9. and effect of permeation permeabilized. Li et al. studied using spectroscopic techniques. Srinivasan and Higuchi 1990. impedance measurements have been used to study the effect of sonophoresis (Mitragotri et al. b). The authors discovered syner- gistic formulations of permeation enhancers where f is the frequency of the applied alternating using this screen. can be performed rapidly and by use of auto- cal gradient. 9. impedance drops finally attaining enhancers (Potts and Francoeur 1993. Also. sage of electricity across skin has been pre. Paliwal et al.4. Z. 1996. Current across skin can thus be mated systems the throughput of impedance-­ related to permeability of skin. P.M. 2004. of using impedance to assess skin permeability is tions has been developed that relates skin imped.1 Theory 1996. since these measurements analogous to diffusion of solutes under a chemi. Impedance-based per- sented by Tregear (Tregear 1966). transform infrared (FTIR) spectroscopy has been The impedance of intact skin is in the range of used extensively in determining skin structure. Karande et al. 2005. Flow of ions across skin under an electric field is and expensive. (2001) as 9. 2006a. several hundred kilo ohms (kΩ). Fourier ute and the solvent in which it is dissolved. 2006). which corresponds to removal Rerek 1998. 2004. faster. Moore and a value of ~1 kΩ. Tezel et al. A formal porous pathway ing a biologically active hormone across the skin theory based on Nernst–Planck flux equations at therapeutically relevant doses. FTIR spectroscopy is used to track solute mol- iontophoresis (Burnette and Bagniefski 1988. Skin and its based assays is significantly larger compared to appendages can be represented by an equivalent diffusion cells. Karande measurements across the SC provide improved Kalia et al. Cropek and P.168 D. is provided by Tang et al. 2001). 2006a. This system. R. in vitro 1 impedance Guided High Throughput (INSIGHT) é 1 ù2 screen.10) Transport of solute molecules in the skin can be where C is dependent on the properties of the sol.9) mulations per day for their ability to modulate ( 2p fC ) úû 2 êë skin permeability. that impedance serves only as a surrogate mea- ance to skin permeability (Lakshminarayanaiah sure of actual skin permeability. hydration.5. Karande et al. The actual flux 1965. Mendelsohn of the entire barrier (Naik et al.3. impedance measurements are relatively simpler. and sensitivity (Dugard and Scheuple 1973). diffusion methods.5. meability assessment provides direct readout of barrier integrity and does not require subsequent 9. 2001. ­ ecules in the epidermis by recording their . of this equivalent skin model can ation enhancers on skin permeability (Karande be represented as et al.

Depth-dependant profiling of the solute penetration. L is the total deflection are a few of the other methods that thickness of skin and D is the effective diffusivity have been suggested in this area (Gotter et al. This review also sheds light on possible reasons pled mass spectroscopy to track permeation of why two particular studies did not observe gold nanoparticles in the rat skin (Sonavane et al. Rossi et al. of the solute in skin.Dn 2p 2 t ö C ( x ) = C0 ç1 . Rocha et al. A compre- cation in vivo (Williams et al.11) as 9. ATR. the solute and hence its permeability across the then. solution for Eq. 2007. (1997). depth x from skin surface and C0 is the concentra. Sonavane hensive review of their findings is provided by et al. ability has been reported (Rougier et al. 2003.5) given as (Pirot et al. b). This limitation to assess barrier integrity of the skin. that the quantitative measure of water loss skin.11) è L ø n =1 np è L ø è L2 ø where C(x) is the concentration of solute at a 2001. A quantita- can. tive correlation between TEWL and skin perme- tion techniques (Naik et al. 1989.å C0 sin ç ÷ exp ç ÷ (9. assess the effect of permeation enhancers on skin pholipids in skin (Xiao et al.5 Trans Epidermal Water Loss æ ¶C ö -D ç ÷ . deriving this correlation that is discussed in Pirot A device. across skin. Other groups have used photoacoustic absorption and TEWL (Tsai et al. have used a combination of UV-vis spec. Several studies have reported the use of TEWL to copy and imaging to track penetration of phos. troscopy. in theory. Remittance spectroscopy and photothermal tion of the solute at the skin surface. Recent literature continues to pro- Carbopol 940 and transdermal gels (Christ et al. 2008). provide a critical comparison of different Singh 2002. The integrated absor. mation in the superficial 1–2 μm layer of the bance of these vibrations is directly propor. Tokudome and Sugibayashi 2004). which the solute diffuses out of the skin can be obtained by differentiation of Eq. (9.5. and Measurements in Transdermal Drug Delivery 169 molecular vibrations. can be used from those of the SC components. that can ade- et al. Chilcott spectroscopy for studying permeation of et al. integration of this rate è ¶x ø x = 0 Mammalian skin has evolved to regulate the with time yields the total amount of solute diffus. (9.9  Models. vide opposing views on the use of TEWL as a . 2005a. quantitative correlation between percutaneous ­ 2008). Levin and Maibach (Levin and Maibach 2005). Further.÷ . This information One of the primary functions of skin is to regu- can be used to obtain the permeability profile of late the loss of water from the body. 2002). skin. It follows. and inductive cou. ing across skin in a given time. Luzardo-Alvarez et al. 1990). 1989) Several other methods have been used to study and is found to be consistently reproducible permeation and absorption of material into and in vivo and in vitro (Pinnagoda et al. Several assumptions have been made in from the skin is indicative of its barrier integrity. Repeated tape stripping can be used to tional to the amount of solute present in the scan successive layers of the skin for solute skin. Williams permeability (Loden 1992. Transport properties for the solute in the skin can be achieved by attenuated total can then be obtained from an unsteady state reflectance (ATR)-FTIR spectroscopy. Methods. Kanikkannan and et al. One downside of using ATR-FTIR quately and accurately measure the water vapor spectroscopy is that the solute to be monitored flux at the skin surface. FTIR spectroscopy generally provides infor. 2001). 1993). The rate per unit area at 2008). and hence the rate of needs to be IR active and have a signature distinct trans-epidermal water loss (TEWL). have used Raman micros. 2001. X-ray spectroscopy. be overcome using spectral correc. such as an evaporimeter. 1997) æ xö ¥ 2 æ np x ö æ . transport of material into and out of the body. Raman spectroscopy techniques and their appli. Xiao et al.

1998). Poelman et al.2 Surface Loss Alternate approach to determine in vivo percuta- neous absorption is to measure the loss of solute 9. surrogate enough. 1984) in vivo by measur- across the skin. ments are considered reliable and superior to measurements made on model systems in vitro. Laser Doppler flowmetry (LDF) has been of a solute across the skin in vivo in much the used to study absorption of prostaglandin E1 same way as diffusion cells with excised skin (PGE1) (Foldvari et al. Caution needs to be exercised when 9. Cropek and P. especially in supersaturated systems. Metabolism can lead to by-products Several of the methods discussed above for that have significantly different pharmacokinetics in vitro skin barrier assessment can be adopted . Generally. 1989).1. Vasoconstriction can filled with drug solution and strapped on to an be used as an endpoint to measure transdermal animal in vivo or the arm of a human volunteer delivery of topically applied corticosteroids (Wurster and Kramer 1961. Karande measure of skin barrier integrity (Fluhr et al. itative than quantitative. 1995.1 Systemic Bioavailability as an endpoint to determine the efficacy of trans- A solute whose percutaneous absorption is to be dermal delivery of insulin (Mitragotri et al. 2006). Nevertheless. tion technique such as radiolabeling with 3H or 14 C is necessary (Wester et al. The downside of this method is concentration measured in blood plasma or urine that it works only for drugs that have a detectable over a period of time. Different endpoints can (Wilkin et al. Wherever possi.2 Pharmacological Response 9.3 Other Approaches using such techniques if the drug is susceptible to metabolism. topically applied nicotinates (Le and Lippold tem allows one to study the passive permeation 1995).6. which in itself can be a poor assumption. Such methods can amount absorbed.M. These amounts generally biological endpoint and can tend to be more qual- tend to be very low and hence a sensitive detec. Netzlaff et al.6  valuation of Skin E from the surface as it penetrates the skin. and pharmacodynamics than the parent com- 2006.1 Diffusion Measurements A good method to assess percutaneous absorp- tion is to measure the biological or pharmaco- Diffusion cells have been designed that can be logical response to the drug. can be recovered.6.170 D. Quisno and Doyle (Barry 1983) and vasodilation as an endpoint for 1983. Leopold and Lippold 1992). assumes that the skin does not act as a reservoir. 2008). ble. 9. Difference In vivo methods for quantification of solute per. Gschwind et al. in vivo measure. and then be used to quantify permeation rates minoxidil (Wester et al.1. methylnicotinate or model membranes. the reservoir can be infinitely large when com- bolic. eral ­commercial devices based on this principle are readily available today for laboratory work. pound and can give misleading results. ing changes in cutaneous microcirculation. 1985. as endpoints as well as faithfully reproduce meta. and practically feasible. measured is applied topically to the skin and its Chen et al. pharmacokinetic. Also.6. 9.6. con- tained in a reservoir. 1983.6. 2006a). Reduction in blood glucose levels has been used 9. Such a sys. this method behavior of the drug molecule. and pharmacodynamic pared to the amount absorbed. the method used to potentially eliminate variables associated with detect the residual amount needs to be sensitive using excised human or animal skin. sev. This Permeability In Vivo can be achieved when all residual material. between concentration or amount at time zero meation across the skin serve as a gold standard and at time of recovery provides an estimate of in transdermal drug delivery.

Tape stripping and TEWL et al. ficient intensity that is distinct from the signa- ping in vivo in healthy human subjects to deter. 2008. is implanted under the dermis where. have used lute amounts of the diffusing solute in different tape stripping to quantify skin penetration of layers of the skin. 2007). For both techniques. 2007). Puglia et al. painful and invasive procedures that are less pop- sively in tracking solute permeation in the skin ular in studying percutaneous absorption of sol- (Ayala-Bravo et al. (Zhai et al. have In addition. 1991). et al. 2008). Kalia et al. straightforward and minimally invasive or nonin. in humans. similar in principle to FTIR spec. the molecule of to determine solute permeability as suggested in interest should have an active IR signature of suf- Eq. (Naik et al. Among the various spectroscopic techniques. Dujardin et al. these are ATR-FTIR spectroscopy has been used exten. in vivo. A . (Herkenne et al.9  Models. A big advantage cellular tissue components by passive diffusion. 2002). The amount of solute that penetrates the Raman spectroscopy to obtain depth profiles of SC can be quantified in vivo. Raman thin probe perfused with a physiological solution spectroscopy. by tape trans-retinol in the epidermis for 10 h after appli- stripping with an appropriate adhesive tape (Tsai cation in an in vivo setting (Pudney et al. Maibach et al. 2003. 2004. 2001). have looked at of successive tape strippings on TEWL in vivo in the effect of surfactant treatment and iontophore- human subjects as well as compared two differ. have demonstrated the use of vasive. In case of animal studies. 2008). technique that can be used in vivo to assess skin taneous absorption in vivo is provided by barrier integrity. it can exchange material with the extra- mal solute penetration in vivo. nique to measure the diffusion of solutes across FTIR spectroscopy and its use in in vivo studies the skin (Kreilgaard 2002. Umemura et al. have used imped- Herkenne et al. Pudney et al. 2005. Stamatas ­modification at all. have used confocal Raman spec- is possible to quantify penetration in deeper skin troscopy to detect dimethyl sulfoxide in the SC layers by sacrificing the animal and harvesting its (Caspers et al. Recently. of Raman spectroscopy is that it does not require The perfusate from the probe can then be ana- tape stripping of the skin for depth profiling and lyzed for the solute diffusing across the skin. Tsai et al. 2006). Curdy utes. 2002). 2003. A comprehensive review of ATR. Curdy et al. Escobar-Chavez et al. 2002). sis on skin impedance in vivo (Kalia and Guy ent configurations for devices measuring TEWL 1997). have documented the effect (Dujardin et al. Impedance spectroscopy is a lipid nanoparticles (Puglia et al. Xhauflaire-­Uhoda et al.8). A review highly sensitive and relatively straightforward of tape-stripping methods in determining percu. describe the use of physical or chemical enhancers of skin permea. ATR- skin. have used tape strip. 2008). Several ance spectroscopy to follow barrier recovery reports have documented the use of TEWL to after the application of iontophoresis (Curdy assess the barrier integrity after treatment with et al. A appears in Naik et al. several other techniques have used TEWL measurements to study barrier repair been utilized to quantify in vivo transdermal after the application of miconazole nitrate and delivery (Herkenne et al. (9. These include tape stripping (Xhauflaire-­ Uhoda et al. 2005). The concentration profiles can then be used FTIR and Raman spectroscopy. An additional drawback mine the pharmacokinetics of topically applied of using Raman spectroscopy is that it can only maxacalcitol from an ointment and lotion provide relative concentrations as against abso- (Umemura et al. 2006). and Measurements in Transdermal Drug Delivery 171 for in vivo measurements with little or no is thus a truly noninvasive technique. have used in vivo confocal Raman micro- measurements in particular are attractive for spectroscopy to study the uptake of vegetable oils in vivo measurements as they are relatively and paraffin oil in infants (Stamatas et al. Mathy et al. tures of skin components. especially in human subjects. et al. has also been used in studying transder. impedance measurements to assess effects of bility (Atrux-Tallau et al. creation of suction blisters and punch biopsies. 2008). 2008). Methods. Although relatively straightforward. on equili- troscopy. Kolli and Banga electroporation on barrier function in vivo in rats 2008). Remane microdialysis has been suggested as a novel tech- et al. it Caspers et al. bration.

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.......... 196 10...........................de perature keratinocytes © Springer-Verlag Berlin Heidelberg 2017 185 N.. Campus E 8......3.1........... H.... Germany Cv Concentration in the vehicle e-mail: ufs@mx.............7 Duration of Exposure and Sampling Period ...........2 I n Vitro Barriers for Skin Absorption Skin Layers ...2 Open Versus Occluded Dosing ..... Germany HaCaT Human adult low calcium high tem- e-mail: Claus-Michael....3...... Schaefer (*) • D.4.2.............................................. 191 10........... 195 10.............. and Claus-Michael Lehr Contents 10...... 197 10.....2.....2 Finite Dosing Studies......................3....1007/978-3-662-53270-6_10 ........ 192 10.............................1 Excised Human Skin........1.............4  esults Obtained from Permeation R 10........... 192 Conclusion..................................2...................2 Rate Determining Processes Involved 10......................9 Influence of Thickness of Skin Studies..... Human Native and Reconstructed Skin Preparations for In Vitro 10 Penetration and Permeation Studies Ulrich F........ 193 10............. 186 10........4 Artificial Skin Surrogates.... 193 References................... 198 10.......3.... 196 10... Dragicevic.)..............2..10 Number of Experiments/Replicates.........5 Temperature ...........6 Selection of Receptor Fluid ....uni-saarland...... the vehicle Saarland University.......... DOI 10.......................-M..........3................3............de EC European Commission S...1........ 196 in Skin Absorption. 186 10..