Advanced Drug Delivery Reviews 65 (2013) 152–168

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Advanced Drug Delivery Reviews

journal homepage: www.elsevier.com/locate/addr

Modeling the human skin barrier — Towards a better understanding of

dermal absorption☆
Owen G. Jepps a, b, Yuri Dancik e, Yuri G. Anissimov a, b, Michael S. Roberts c, d,⁎
a
Griffith University, School of Biomolecular and Physical Sciences, Brisbane, QLD, Australia
b
Griffith University, Queensland Micro- and Nanotechnology Centre, Brisbane, QLD, Australia
c
The University of Queensland, School of Medicine, Therapeutics Research Centre, Brisbane, QLD, Australia
d
The University of South Australia, Therapeutics Research Centre, School of Pharmacy and Medical Science, Adelaide, SA, Australia
e
Proctor & Gamble Eurocor, Temselaan 100, Strombeek-Bever, Belgium

a r t i c l e i n f o a b s t r a c t

Article history: Many drugs are presently delivered through the skin from products developed for topical and transdermal
Accepted 9 April 2012 applications. Underpinning these technologies are the interactions between the drug, product and skin that
Available online 14 April 2012 define drug penetration, distribution, and elimination in and through the skin. Most work has been focused
on modeling transport of drugs through the stratum corneum, the outermost skin layer widely recognized as
Keywords:
presenting the rate-determining step for the penetration of most compounds. However, a growing body of
Modeling
Human skin
literature is dedicated to considering the influence of the rest of the skin on drug penetration and distribu-
Barrier tion. In this article we review how our understanding of skin physiology and the experimentally observed
Dermal absorption mechanisms of transdermal drug transport inform the current models of drug penetration and distribution
Transdermal drug transport in the skin. Our focus is on models that have been developed to describe particular phenomena observed
Transdermal penetration at particular sites of the skin, reflecting the most recent directions of investigation.
Distribution © 2012 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
2. Overview of skin transport modeling. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
3. Contributions to drug penetration and distribution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
3.1. Stratum corneum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
3.1.1. Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
3.1.2. Mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
3.1.3. Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
3.2. Viable epidermis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
3.2.1. Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
3.2.2. Mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
3.2.3. Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
3.3. Dermis and deeper layers. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
3.3.1. Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
3.3.2. Mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
3.3.3. Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
3.4. Appendages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
3.4.1. Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
3.4.2. Mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
3.4.3. Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
4. Global perspective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160

☆ This review is part of the Advanced Drug Delivery Reviews theme issue on “Modeling the human skin barrier — Towards a better understanding of dermal absorption”.
⁎ Corresponding author at: Therapeutics Research Centre, School of Medicine, University of Queensland, Princess Alexandra, Hospital, Brisbane, QLD 4120, Australia. Fax: + 61 7
3240 5806.
E-mail address: m.roberts@uq.edu.au (M.S. Roberts).

0169-409X/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
doi:10.1016/j.addr.2012.04.003
 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168 153

4.1. Overall distribution models . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160

4.2. Prodrugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
4.3. In vivo vs. in vitro results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
4.4. Clinical response . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
5. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
Appendix A. Summary of equations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
165

1. Introduction Drug transport through the skin is primarily considered to be by

Transdermal drug delivery is an important delivery mode. Over
one third of drugs under clinical evaluation involve delivery into or
via the skin [1]: this figure includes both drugs whose sites of action
are in the skin, and those meant to be taken up by the systemic circu-
lation. The advantages of transdermal delivery include the avoidance
of the pain, possible infection and compliance issues related to injec-
tions; the possibility to control drug release over long periods; and
avoiding the first-pass metabolism via the oral route that can lead
to large loss of drug [2,3]. The challenges include the significant vari-
ability in penetration between people [4], and between sites on an
individual [4]; and the considerable effectiveness of the skin barrier
(particularly its uppermost layer, the stratum corneum) in limiting
drug penetration. Predicting the permeability of a given compound
through the skin is in general quite difficult, due to the highly com-
plex nature of the structures and mechanisms that constitute the de-
livery pathway.
In this article we will review the current understanding of the
structures and mechanisms that most affect transdermal drug deliv-
ery and the distribution of drugs within the skin. Our focus will be
on the transport and distribution of the solute drugs themselves:
we will not consider the delivery of nanoparticles or vesicles, which
has been reviewed elsewhere [5,6]. We begin with a brief overview
of the skin transport processes (that we will consider in more detail
in Section 3 from a physiological perspective), and consider the
main quantities and mathematical representations. We then investi-
gate in greater detail the various contributions to this overall picture.
Given the importance of skin physiology in determining drug transport
and distribution, we group these contributions according to skin mor-
phology (the stratum corneum; viable epidermis; dermis and deeper
layers; and appendages). In each section we will consider the various
transport mechanisms that arise, how their effects are modeled, and
their overall contribution to percutaneous penetration. We then consid-
er how these contributions are combined to form comprehensive pen-
etration and distribution models, as well as other important topics,
before concluding.
2. Overview of skin transport modeling
Drugs applied to the skin surface enter the skin through either the
uppermost layer of the epidermis – the Stratum Corneum (SC) – or
one of the appendages (including hair follicles and ducts leading to
eccrine sweat glands) [7,8] (Fig. 1). Passage through the SC leads to a
further series of skin layers — the viable epidermis, the dermis and the
hypodermis/subcutaneous tissue. Permeation into the vascularized
dermis (which also houses the lymphatic pathways) and deeper tissues
offers the possibility of systemic drug distribution. The heavily vascular-
ized nature of the hair follicle and appendageal glands also suggests the
possibility of systemic drug distribution via those routes. While the SC
barrier, which serves as the major rate-limiting component to skin pen-
etration for most drugs, has been the focus of most penetration models,
much less is known about transport beyond the SC, and about transport
via the appendageal routes.
diffusion — the ‘random walk’ of drug molecules through the various
structures within the skin. This walk can be mediated by active trans-
port (whereby protein transporters facilitate drug transport in certain
environments) and convective transport (whereby molecules are
driven by local currents in the lymphatic or vascular systems or inter-
stitial spaces). Uptake into the lymphatic or vascular systems impacts
upon drug distribution, as well as facilitating drug clearance. Apart
from clearance, drug may also be effectively removed through metab-
olism in the skin.
When modeling skin transport processes quantitatively, we typi-
cally consider the various physiological regions of interest (potential-
ly including skin and deeper tissue layers, appendages, the circulation
and other tissues) as compartments. Drug levels within the compart-
ment can be modeled as a single time-dependent value c(t) (the
traditional compartment model) or as function c(x,t) of both position
within that compartment (usually skin depth) as well as time. For the
latter choice, drug transport within the compartment may be mod-
eled using partial differential equations that describe the effects of
drug diffusion, convection, elimination and metabolism. Drug trans-
port between compartments, as well as drug elimination or metabo-
lism, can be described using rate constants. For a comprehensive
review of mathematical models related to transdermal drug delivery,
we refer the reader to Ref. [9].
One of the principles behind such models is that the parameters
describing the various transport processes – diffusion, convection,
elimination, etc. – are expected to depend on the drug under investi-
gation, but to remain independent of the dose regimen. From a
mathematical viewpoint it is easiest to establish model parameters
for drug delivery processes that have reached a (pseudo) steady
state — that is, a state where drug concentrations in the various com-
partments remain essentially constant (until, for example, the deple-
tion of the administered dose of drug becomes significant). Once
these model parameters have been determined for a dose regimen
that produces a steady state, the same model parameters can then
be used to explore the drug transport and distribution behavior for
other doses.
Quantities that can be established experimentally, and that relate
directly to the drug transport and distribution in skin, include the
steady-state flux of drug JSS into the skin (and related quantities
such as the permeability coefficient kp, which represents the constant
of proportionality between the steady-state flux and drug concentra-
tion C in the applied dose, i.e. JSS = kpC; and the maximum steady-
state flux Jmax, obtained when C is maximal, and thus equal to the
solubility S of the drug in the applied vehicle, i.e. Jmax = kpS); the lag
time between drug application and attainment of the steady state;
the clearance of drug through excretion; and drug concentrations in
the various skin layers, circulation or other tissues. We note that
this final set has traditionally only been ascertained invasively, al-
though modern microscopy techniques now allow the possibility of
in situ concentration estimation [10–12].
The influence of specific molecular-scale mechanisms (such as ac-
tive transport) on the overall transdermal transport of a drug is often
contained within these compartment-scale model parameters. As un-
derstanding of these processes improves, the sophistication of models
154 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168
Fig. 1. Schematic overview of the skin layers, appendages, blood and lymphatic vessels.
used to describe drug penetration and distribution increases, as does
our understanding of the dependence of model parameters on drug
properties. This in turn facilitates better prediction of transdermal
drug transport, which is the ultimate aim of such work.
3. Contributions to drug penetration and distribution
Given the long-established importance of the SC as the main pen-
etration barrier for many drugs [13], it is important to recognize that
the skin layers below the SC can also play important roles in the pen-
etration and distribution of topically administered drugs. Lower skin
layers are important, for example, when considering the delivery of
lipophilic drugs, or drugs targeting the dermis, deeper tissues, or
the circulation. In this section, we consider the various layers of the
skin and the appendages, outlining the local transport effects that im-
pact upon drug penetration and distribution, and the way in which
they can be modeled.
3.1. Stratum corneum
3.1.1. Morphology
The SC is the outermost layer of the epidermis. Cells migrate from
the dermal–epidermal junction at the base of the epidermis to the SC
over a period of approximately 2 weeks during a process known as
desquamation [14]. The final stages of desquamation involve the dif-
ferentiation of epidermal keratinocytes into flat, closely packed inter-
digitated corneocytes, embedded in a highly organized, dense lipid
matrix [15,16] that is shed from the differentiating keratinocytes.
The corneocytes comprise a dense scaffolding network of keratin fila-
ments believed to support the corneocyte shape; a range of hygro-
scopic compounds collectively termed Natural Moisturizing Factor
(NMF) that help maintain SC hydration; and water-insoluble cell
walls made up of highly cross-linked proteins including loricrin,
involucrin, and filagrin. The intercellular lipid matrix chiefly com-
prises a mixture of ceramides, cholesterol, triglycerides and fatty
acids, arranged into lipid lamellae. The relative proportions of these
compounds and their precise physical structure are matters of ongo-
ing research (cf, for example, [16,17]).
The lower epidermal layers are thus usually referred to as the via-
ble epidermis, in contradistinction to the cells of the SC that are enu-
cleated during differentiation [18,19]. This ‘bricks and mortar’ [20]
structure is primarily responsible for the SC's effectiveness as the
main physicochemical barrier, both against the permeation of exoge-
nous compound into the skin, as well as endogenous compounds out
of the skin. It is worth noting that imperfections in the SC structure
(such as within the cell structure, or lacunae of water found within
the lipid layers [21]) are likely to impact upon transport properties,
although it is not clear to what extent.
3.1.2. Mechanisms
Transport across the SC is essentially by passive diffusion, whether
it be via an intercellular (i.e. restricted to the lipid matrix) or transcel-
lular (i.e. through both corneocytes and lipid matrix) pathway. Perme-
ation along such pathways is thus contingent upon a compound's
affinity with lipid environments, with the internal environment of
the corneocyte, and upon the capacity of a molecule to permeate the
corneocyte cell wall. The relative importance of these pathways has re-
cently become a renewed focus of investigation, both experimentally
and from theoretical considerations. Despite initial support for the
transcellular route [22], early experimental studies indicated that the
intercellular lipid pathway was the dominant for SC permeation
[23–26]. More recent microscopy evidence supports the view that the
transcellular route is an important one [27], as do results from recent
SC transport models (Section 3.1.3). Scheuplein suggested different
pathways for molecules, depending on their polarity [28], a view sup-
ported by early experimental evidence [29,30].
 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168
3.1.3. Modeling
One of the simplest mathematical models of drug transport
through the SC can be formulated as a very large well-stirred donor
phase applied to the top of SC (that is initially devoid of drug), with
rapid clearance at the lower SC boundary (for example, due to the
phase beneath the SC being maintained at an effective zero drug con-
centration). If the SC is represented as a homogeneous membrane,
transport can be described by the diffusion equation (Fick's 2nd law)
∂C ðx; t Þ ∂2 C ðx; t Þ
¼ DSC
∂t ∂x2
together with the initial and boundary conditions:
C ðx; 0Þ ¼ 0; C ð0; t Þ ¼ K SC C d ; C ðhSC ; t Þ ¼ 0
where C(x, t) is the concentration at depth x and time t, D is the diffu-
sion coefficient, KSC is the partitioning coefficient, Cd is the concentra-
tion in the donor phase and hSC is the thickness of the SC. The solution
to Eq. (1.1) with initial and boundary conditions given by Eq. (1.2) is
most commonly expressed as the amount of solute penetrating SC (Q)
vs. time (t) [31]:
!!
DSC 1 2X ∞
ð−1Þn D 2 2
Q ðt Þ ¼ K SC AC d hSC t− − exp −t 2SC π n
2
hSC 6 π n¼1 n2 hSC
where A is the area of solute application to SC. The summation term in
2
this equation can be ignored at long times (practically at t > hSC /(2D))
yielding the equation that is most commonly applied to experimental
data in the transdermal penetration literature:
! binding of solute molecules to soluble proteins may have sequestered
K D AC h2  
Q ðt Þ ¼ SC SC d t− SC ¼ kp AC d t−t lag
hSC 6DSC
where
2
K SC DSC h sequestration of drugs in the VE [41]. It has been recently suggested
kp ¼ and t lag ¼ SC :
hSC 6DSC
While it has been formulated here as the solution to diffusion
through a homogeneous membrane with simple boundary condi-
tions, Eq. (1.4) is widely applied because its form often remains
unchanged for more sophisticated SC models, with only equations
for kp and tlag modified [32–34]. Indeed, the difficulties in determining
the actual path taken by different drugs through the SC are largely re-
lated to this observation. These other more complex models are
reviewed in [9]. Solutions to various application scenarios (finite ve-
hicle volume, finite time exposure with wash off and evaporation
from the vehicle) are also presented in Ref. [35].
Models that explicitly include the structure of the SC facilitate the
comparison of model steady-state and transient lag times with exper-
imental values, providing valuable insight into the importance of the
intercellular [36] and transcellular [37,38] routes. A similar approach
has provided evidence that corneocyte uptake and/or slow binding in
the SC influence permeant transport behavior, even for simple mole-
cules such as water [34,39,40].
An important phenomenon of drug delivery through the SC is the
so-called ‘reservoir effect’, where lipophilic compounds are effective-
ly sequestered by the SC. Such compounds partition preferentially
into the lipophilic SC rather than the aqueous viable tissues — the
SC–VE boundary provides a significant energy barrier for such mole-
cules. However, a compound must be sufficiently diffusive within
the SC for an effective reservoir to be formed. For a recent review of
the reservoir effect, the reader is referred to Ref. [41].
155
3.2. Viable epidermis
3.2.1. Morphology
The viable epidermis (VE) is an avascular environment, consisting
mainly of keratinocytes and comprised of approximately 40% protein,
40% water and 15–20% lipids [19,42,43]. The undulating epidermal–
dermal junction consists of papilla that project into the dermis [19] —
cells in the basal layer of the epidermis form the most important struc-
tural and functional link to the dermis below [43]. We recall that
the epidermal cells undergo continual migration from the epidermal–
ð1:1Þ dermal junction up toward the SC during desquamation.
3.2.2. Mechanisms
Drug diffusion in the VE can be considered as diffusion through an
ð1:2Þ aqueous medium that is hindered by the significant presence of pro-
teins and punctuated by cell membrane crossings. The high fraction of
water present in the VE makes this region a more effective barrier
against lipophilic compounds, which have greater affinity for non-
polar environments [13]. Wenkers and Lippold suggested that the
VE provides a decisive barrier to the penetration of NSAIDs from a
lipophilic vehicle, based on correlations between skin permeability
and octanol–vehicle and PBS–vehicle partition coefficients [44].
Other VE processes that impact upon the penetration and distribu-
tion of drug within skin include drug binding and sequestration within
ð1:3Þ the VE, drug metabolism and active transport. The detailed influence of
drug binding, sequestration and metabolism on drug penetration and
distribution is not always well established, although it is generally
understood that they can limit the extent to which drug is available to
permeate into lower regions. Bhatt et al. observed higher than expected
amounts of the lipophilic pesticide tecnazene remaining in skin tissue
after a 48 h in vitro skin penetration experiment, suggesting that the
them within the tissue [45].
ð1:4Þ This effect can be represented in terms of a drug's bioavailability:
that fraction of drug that is available at a particular site for a particular
process (such as permeation into a neighboring region). Experimental
evidence suggests that epidermal metabolism occurs in the basal
layer [46,47], and that metabolism can play an important role in the
ð1:5Þ
that the presence of keratin in the VE may affect diffusion into the
VE and/or partitioning from the SC into the VE [48].
There is a wealth of information regarding carrier-mediated trans-
porters in human skin [49], and studies of active transport in the VE
center on those transport proteins expressed in epidermal keratino-
cytes. However, studies of the kinetics associated with the active
transport of drugs – either for enhancing permeation or for serving
as a barrier – have yet to appear in the literature.
3.2.3. Modeling
The barrier effects of the VE can be incorporated into skin perme-
ation models by treating the VE as a homogeneous unstirred aqueous
layer immediately below the SC. Thus the SC and VE form two barriers
in series: the total epidermal permeability is calculated as
1 1 1
¼ þ ð1:6Þ
kp;epi kp;SC kp;VE
in accordance with solutions to the diffusion equation [32,50,51] —
see Ref. [9] for more details.
Cleek and Bunge compared such a model to single-membrane
models (finite or semi-infinite) representing the SC only, to further in-
vestigate the role of the VE [52]. By defining the ratio B= kp,SC/kp,VE,
they could consider the overall contribution of the viable epidermal bar-
rier in terms of its effectiveness relative to the SC barrier, obtained
through formal solutions to the diffusion equation for each choice of bar-
rier. The quantity B can be related to drug physicochemical properties
156 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168
such as lipophilicity [53], which are expected to impact upon the relative
permeability of drug in the lipophilic SC and drug in the aqueous VE. At
penetration times that are sensitive to the existence of both barriers,
the two-membrane model shows that penetration into the skin is depen-
dent on B and therefore on drug lipophilicity: for highly hydrophilic
drugs (Bb 0.01) penetration is limited primarily by the SC, while for in-
creasingly lipophilic drugs the effects of the aqueous layer become
more significant, “choking” the flux of such drugs into the skin in agree-
ment with experimental observations [44]. Such models can also be used
to correct lag time predictions [52], and various methods have been
proposed for estimating the parameter B [53], from which one can gen-
erate corrections to the SC permeability predictive model of Potts &
Guy that take into account epidermal resistance [52,53]. More recently
this model has been extended to account for ionization and vehicle
properties, applied to model the penetration of lipophilic (N-nitroso-N-
methyldodecylamine, NDOMA) and hydrophilic (hydroquinone) per-
meants through skin [54] and incorporated into models of skin diffusion
which include the effect of desquamation [55,56].
Such two-layer models have been used to estimate the permeabil-
ity of glucose in the viable epidermis from tape stripping data [57].
Glucose permeability was calculated to be 38% of that in dermis, com-
pared with a value of 30% of the dermal permeability that was
obtained from a side-by-side diffusion cell experiment on human epi-
dermal membrane.
Drug binding and sequestration in the viable epidermis affect the
availability of drug for permeation into the dermis and beyond. The
steady-state concentration of drug cSS at a site can be determined by
balancing the input flux of drug JSSFavailableA (for input flux JSS through
area A, where fraction Favailable of drug is available at the site) with the
clearance flux cSSCl (for clearance rate Cl):
kp;SC cV Favailable A J SS Favailable A
cSS ¼ ¼ :
Cl Cl
However, drug binding or sequestration reduces the flux incident
at lower levels. Siddiqui et al. proposed that epidermal sequestration
of lipophilic solutes with steroid accumulation in the VE reduces the
steady-state flux to JSS = kp, SC(cV − cSS) [27], whence
kp;SC cV Favailable A
cSS ¼
Cl þ kp;SC Favailable A
leading to a decrease of up to 30% in the predicted steady-state der-
mal steroid concentration.
The relationship between drug binding in the VE and dermis and
drug physicochemical properties is not generally understood: while
correlations with drug lipophilicity have been established [58], such
relations are not universally predictive across families of compounds
[59]. Recent studies show that once a lipophilic compound has per-
meated beyond the SC, it can bind with soluble proteins and undergo
bound transport [60]; and correlations have been recently established
between lipophilicity and preferential distribution in the VE [61],
although not necessarily in the dermis as well [62].
The effects of metabolism are generally incorporated into trans-
port models by including terms to represent the fraction of available
drug that remains unmetabolized. A more sophisticated approach
was adopted by Sugibayashi et al. in their study of the metabolism
of ethyl nicotinate (EN) into nicotinic acid (NA) by the enzyme ester-
ase [63]. Models consistent with their experimental data indicate that
enzyme distribution within the skin had little effect on the overall
flux of EN and NA, but had a significant influence on the concentra-
tion gradients. The relatively poor permeation of drugs whose tissue
residence time is long compared with the metabolic half-life (indicat-
ing that a large amount of drug is metabolized in the time taken to
cross the tissue) has been observed experimentally and modeled by
Boderke et al. [64].
Despite the importance of and recent interest in VE protein trans-
porters, most studies have focussed on the nature of such transporters
rather than on characterizing their effect on drug permeability. Li et al.
[65] showed that the penetration of the NSAIDs flurbiprofen and indo-
methacin is governed by pH- and ATP-dependent saturable kinetics.
Passive diffusion and reversible kinetics alone cannot account for this:
the authors propose active transport as a potential explanation, possibly
by MRP, MCT, OATP or OCTN xenobiotic transporters [65].
3.3. Dermis and deeper layers
3.3.1. Morphology
Immediately beneath the epidermis, the dermis is the thickest
component of the skin, up to 4 mm in depth. Its upper layer, the
100–200 μm thick papillary dermis, consists of thin collagen bundles,
elastin fibers, fibrocytes and ground substance comprising mainly
water, electrolytes, plasma proteins and polysaccharides–polypeptide
complexes [43]. Below this layer is the reticular dermis, made up pre-
dominantly of thick collagen bundles and coarse elastic fibers [43].
Protected by the epidermal barrier, the dermis houses the blood ves-
sels, lymphatics and nervous system within the skin, as well as the
various skin appendages. Below the reticular dermis lies the hypoder-
mis (subcutaneous fat tissue), which may have a thickness of up to
several millimeters. Comprising fat microlobules and fibrous collagen,
it also houses the blood vessels, lymphatics and nerves, and serves to
store energy, insulate the skin from below, and to connect the skin to
underlying structures such as muscle and bone while allowing move-
ment over them [42,43,66].
The structure and organization of the dermal blood vessels have
been described by Braverman [67,68], with models of vessel distribu-
tion with depth subsequently developed by Cevc and Vierl [69]. Most
of the microvasculature is concentrated in the papillary dermis: the
ð1:7Þ
number of blood vessels decreases some 20-fold in the first millime-
ter of dermis; blood vessel area increases with depth from the epider-
mal junction, peaking toward the lower end of the papillary dermis
before decreasing quasi-exponentially. The orientation of vessels
also has a particular structure, with a dense plexus of vessels running
along the epidermal–dermal junction, another less dense plexus in
the reticular dermis, and vessels between the two plexuses that also
connect to the skin appendages (Fig. 1). The system of lymphatic ves-
ð1:8Þ
sels in the dermis runs roughly parallel to the dermal vasculature
structure [43,70].
3.3.2. Mechanisms
Similar to the VE, drug diffusion through the dermis is primarily a
hindered diffusion through an aqueous medium. However, there are
important differences: the vascularization of the dermis contributes
significantly to drug transport and distribution in the skin, as does
the lymphatic system; the dermis is predominantly acellular in na-
ture; and the dermis provides many opportunities for the binding
and sequestration of drug.
The co-administration of vasoconstricting or vasodilating com-
pounds (which alter drug uptake in the vasculature) during penetration
studies has helped shed further light on the role of the vasculature in
drug penetration and distribution [71–81]. In theory, vasodilation
should allow greater clearance of drug into the vasculature, thereby
lowering steady-state concentrations: vasoconstriction should have
the opposite effect. Such behavior has been observed experimentally
for lidocaine delivery together with tolazoline (a vasodilator) or norepi-
nephrine (a vasoconstrictor) [71,72], for salicylic acid and lidocaine
with phenylephrine (a vasoconstrictor), for flurbiprofen with epineph-
rine (a vasoconstrictor) [76] and for antipyrine with phenylephrine
[79,81]. A number of microdialysis studies have also confirmed these
observations [74,77,78,80]. Increased drug distribution at contra-
lateral sites due to vasodilation [75], and reduced distribution at contra-
lateral sites due to vasoconstriction [81] have also been observed.
 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168 157

Vasoconstriction has also been observed to augment deeper tissue

(muscle) penetration of drugs inversely to their fraction unbound in
viable skin (which reflects their distribution in muscle) [79].
There is also evidence to suggest that the orientation of blood ves-
sels plays a role in producing a “convective force” for drug transport
into deep tissue, beyond levels that can be explained by systemic de-
livery alone [82]. Such results have been observed for piroxicam de-
livery into deep tissues both at the site of application [83] and away
from the site of application [84], as well as for the diazepram applied
dermally to perfused rat hindlimb [85].
Lymphatic clearance plays an important role in the removal of
macromolecules [70,86–90], which suggests its potential relevance
to drug transport where drug binding occurs.
While transporter proteins are known to be expressed in the dermis
[91], few studies have been undertaken to date on the role of cutaneous
carrier proteins in drug distribution in dermis. Published studies show,
however, that such transport mechanisms may occur and could be
harnessed for the purposes of drug delivery to the deeper tissues or
the systemic circulation. P-Glycoprotein is a transporter protein belong-
ing to the ABC family and mainly expressed in the dermal components
of human skin (including sweat ducts) and muscle tissue [92]. Ito et al.
showed that P-Glycoprotein can transport itraconazole within dermal
tissue [93]. Another study by the same group showed that the Organic
Anion Transporter 2 (OAT 2) was involved in the transport of topically
applied flurbiprofen in an in vitro experiment [94]. Lipophilicity may
play a role in such transport mechanisms. While the transport of the
lipophilic flurbiprofen was vectorial (greater from the epidermal to
the hypodermal side then in the opposite direction) and saturable, no
carrier-mediated transport of mannitol, a hydrophilic permeant, was
observed.

3.3.3. Modeling
In the same way that the SC and VE can be treated as “diffusion re-
sistances in series” (where the diffusive resistance is given by the in-
verse permeability), Eq. (1.6) can be extended to include the dermis
as a third diffusive resistance. Using such a model, Scheuplein and
Blank found that while the permeability of a homologous family of al-
cohols decreased significantly with increasing number of carbon
atoms n, it remained approximately constant for the dermis [13]
(for n > 6, when the main barrier is provided by the SC, the dermal
and epidermal permeabilities are similar in value).
Much of dermal transport modeling concerns the quantification of
Fig. 2. The two-compartment model in Ref. [90]. (a) Schematic for Qp and Qt represent-
properties related to the vasculature and lymphatic clearance. Typi-
ing the perfusate flow rate into and blood flow rate out of tissue respectively; ka and ke
cally, this is achieved via compartment models where a single value represent drug absorption and elimination rates, (b) observed data for diclofenac
represents the mean local concentration of the drug. For example, remaining in dermal cell with time for a dextran and albumin perfusate, (c) observed
Roberts and Cross [90,95] solve a two-compartment model (Fig. 2) data for diclofenac appearing in perfusate from tissue with time after delivery via a der-
analytically to establish the time-dependent cumulative amount of mal cell for a dextran and albumin perfusate.

drug having reached the dermis. By considering the fractions of

drug unbound in tissue and in the vasculature, one may estimate
the effective drug volume of distribution, which then permits an esti-
mate of the elimination rate constant. From such an approach, impor-
tant clinical parameters such as fractions of sequestered drug and the
solute retention half-life can be estimated. This approach has been
validated with diclofenac absorption data [95].
More complex compartment models that explicitly include the
systemic circulation and distribution into deeper tissues have also
been developed by Singh and Roberts (Fig. 3) [96,97] and by Higaki
et al. [79]. By fitting physiologically based parameters in their model
to match experimental data of the distribution of salicylic acid after
application to dermis in anesthetized rats, the relative importance
of the tissue and vasculature transport to the distribution of drug in
muscles could be examined. In particular, the importance of systemic
circulation (through measuring delivery at contralateral sites) could
be quantified, revealing the dominance of tissue transport for deep
drug delivery at the site of application [96]. This model also describes
the concentration–depth profiles observed during vasoconstriction
[73]. Higaki et al.'s model (Fig. 4) applied to the delivery of 6 drugs
led to similar conclusions at early times, but more significant contri-
butions from systemic circulation at longer times [79], where the
transport rate from viable skin to muscle was found to be significantly
correlated to the fraction unbound in viable skin. The removal of VE in
Singh and Roberts' study may explain to some degree the long-time
discrepancy between these studies.
Despite offering valuable insights, a potential drawback of such
compartment models is the implicit homogeneity, which belies the
expected variation in drug concentration and other effects. This can
be overcome by explicitly building such variation, where known,
into spatially-sensitive models. For example, the variation in vascu-
larization through the papillary and reticular dermis can be incorpo-
rated using the partial differential equation [69]
∂cðd; t Þ ∂2 cðd; t Þ
¼ DðdÞ −kd P ðdÞcðd; t Þ ð1:9Þ
∂t ∂2 t
158 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168

Fig. 3. Drug distribution in the skin layers and circulation described by the model of Singh et al. [96]. (a) Physiological pharmacokinetic model: distribution is measured as concen-
trations c; model parameters include clearances Cl, blood flow rates Q and volumes V; subscripts refer to relevant site. (b) Experimental (data points) and predicted (solid line)
dermal concentration–time profiles in underlying tissue after dermal application of salicylic acid. The first peak is due to direct penetration from topical application. (c) Experimental
(data points) and predicted (solid line) concentration–time profiles in subcutaneous and lower tissues below the applied site. The second peak at about 10 h is attributed to systemic
distribution as well as some direct penetration of salicylate.

where the diffusion coefficient D and elimination kpP are functions
of depth. Such depth-dependence can then be estimated by fitting
against concentration–depth profiles [69].
It is clear from experimental data with vasodilators and vasocon-
strictors that convective capillary transport, while responsible for
clearance of the drug, also contributes to transport of solutes to dee-
per tissues. Recent models [98,99] suggest that transport to lower
layers occurs mostly by blood/lymphatic flow rather than molecular
diffusion alone, as assumed previously. This blood/lymphatic trans-
port can be modeled by replacing the dermal diffusion coefficient
with an effective dispersion coefficient (similar to modeling of liver
clearance by dispersion model [100]). As in the liver model, the dis-
persion coefficient is approximated to be a constant parameter,
independent of other physiological parameters of the dermis. The
dispersion coefficient for the liver model was later related to the
liver morphology and physiologically related parameters [101] — a
similar relationship remains to be established for the dermal trans-
port model.
Kretsos et al. analyzed experimental data on the in vivo penetra-
tion of salicylic acid into de-epidermized rat skin using a distributed
diffusion-clearance model describing transient distribution in whole
skin [102]. Their main goal was to characterize dermal transport of
salicylic acid (SA) by deriving values for the effective diffusion and
partition coefficients (with respect to an aqueous solution at a given
pH) in dermis, and for a volumetric first-order rate constant repre-
senting systemic clearance. The study shows that owing to a balance
between diffusion within and volumetric clearance from the dermis,
the SA concentration–depth profile exhibits an exponential decay
 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168 159

Fig. 4. (a) Compartment model of Higaki et al. [79] describing kinetics of topically applied drugs. Distribution is measured as concentrations C; model parameters include clearances
Cl and volumes V; subscripts refer to relevant site. (b) Solid lines describe predicted intradermal kinetics of diclofenac after topical application on the stripped skin in vivo with the
symbols representing donor cell (△), viable skin (▲), muscle (◊), plasma (♦) and contralateral muscle (●). (c) Predicted diclofenac concentrations in muscle — the total concen-
tration (solid line), the concentration due to direct penetration (dashed line), and the concentration due to systemic circulation (dotted line).

profile. While capillary clearance significantly impacts drug distribu-
tion in dermis, the in vivo dermal diffusion coefficient of SA was
found to be approximately equal to the molecular diffusion coefficient
of SA in dermis, implying that the dermal vasculature does not signif-
icantly increase transport by convection.
Kretsos et al. analyzed in vitro dermal penetration data and devel-
oped the most detailed empirical expressions to date for the partition
and diffusion coefficients in dermis [103]. The models incorporate per-
meant ionization, volume exclusion due to the presence of a fiber phase,
a lipid phase and binding to albumin present in a portion of the dermis'
aqueous phase. A “binding factor” is derived and serves to modify the
values of solute concentration, and partition and diffusion coefficients
of unbound solute based on these considerations. This factor is [103]
KP the permeant's octanol–water partition coefficient. A detailed de-
scription of Kasting and coworker's correlations (as well as their use
in a general skin penetration model) is presented in Ref. [104]. In gener-
al, protein binding in the skin is often approximated by binding to albu-
min since this is the most common protein found in the interstitial fluid.
However, other lipoprotein, α1-glycoprotein [105] and keratin should
also be considered. Evidence of permeation rate differing as a result of
differential protein binding is given by Weiss et al. [106].
Questions that remain to be answered in the study of deep tissue
penetration include [107]: is direct penetration or systemic circula-
tion more important for the delivery of certain drugs to the deep
skin tissue?; and why is the increase in the fraction unbound in viable
skin not important for systemic absorption of certain drugs?

ϕa 3.4. Appendages
Binding factor ¼ ð1−ϕa Þ þ þ φlip fnon K P ð1:10Þ
fu
3.4.1. Morphology
where ϕa and ϕlip denote the albumin and lipid phase volume fractions The conical entrance into the hair follicle from the skin surface
in dermis, fu and fnon fractions of unbound and non-ionized solute, and (known as the infundibulum) extends approximately 500 μm deep
160 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168
(i.e. well beyond the typical depth of the epidermal–dermis junction)
to the sebaceous duct that connects the follicle with the sebaceous
gland. Sebaceous glands are also found independently of hair follicles,
mainly on the face [108]. This gland produces sebum, a lipophilic sub-
stance comprising triglycerides, wax esters, squalene, cholesterol and
its esters [109] that fills the infundibulum and serves various roles
(including protection against bacteria, skin over-wetting, heat loss,
transporting antioxidants to the skin surface, and maintaining skin
integrity) [43,110]. Sebum takes approximately 8 h to pass from the
sebaceous duct to the skin surface [111]. The epithelial wall of the in-
fundibulum is a continuation of the SC whose thickness diminishes
down the follicle [13,66,112]. Below the sebaceous duct, the hair
follicle comprises a series of tightly packed cell layers – an acellular
membrane, the outer root sheath (a continuation of the viable
epidermis), and the inner root sheath – that surround the hair shaft
[43,66,113,114]. At the base of the follicle lies the hair bulb, a region
that contains the dermal papilla and is instrumental for follicular
growth. The follicle is heavily vascularized: the bulb is surrounded
by interconnected blood vessels running parallel to the length of the
follicle; vessels run parallel to the follicle above the bulb, connecting
with the vessels surrounding the sebaceous gland (whose acini are
also highly vascularized) [115].
Eccrine sweat glands produce sweat to help cool the body by
evaporation, improve grip, and sensitize skin [42,43,116]. The glands
are housed in the lower dermis or hypodermis, and connected to the
surface by a duct approximately 100 μm in diameter that coils about
the gland before rising straight through the dermis and spiralling up
the epidermis [117]. Water, salts and electrolytes can be reabsorbed
through the duct walls to maintain homeostasis as required [116].
The skin also contains apocrine and apoeccrine glands that resemble
eccrine glands. In adults, apocrine ducts connect to the infundibulum,
and their precise product (and its function) has not been isolated
[43,66]. Apoeccrine glands appear during adolescence, and share fea-
tures of both eccrine and apoeccrine glands [118,119]. These eccrine
glands and ducts are also highly vascularized, as are the apocrine
glands [115,120].
3.4.2. Mechanisms
The skin appendages present an attractive alternative pathway for
drug delivery into the skin, raising the possibility of avoiding the chal-
lenges of delivering drug through the SC. Lipophilic sebum provides
a natural pathway for the diffusion of lipophilic drugs into the hair
follicles and sebaceous glands, but presents a barrier for hydrophilic
drugs [66]. While this behavior has been observed experimentally,
the potential for drugs to pass from the infundibulum into deeper
parts of the follicle/sebaceous gland, and ultimately to permeate into
the vasculature or the skin, remains to be understood. In a similar
fashion, the potential for drug penetration via the eccrine or apocrine
ducts has yet to be understood, although recent work indicates this pos-
sibility [119].
A clear understanding of the mechanisms promoting or retarding
drug transport via the hair follicles and glands remains to be estab-
lished, despite the wealth of experimental studies.
3.4.3. Modeling
Scheuplein and Blank identified the importance of the appenda-
geal pathway during the initial stages after topical drug application
[13,121]. This work has been much more recently verified using mi-
croscopy techniques [122,123].
It appears that lipophilic vehicles such as liposomal formulations or
nanoemulsions facilitate drug penetration into deeper tissues by improv-
ing permeation through the follicular pathway, in hairy rats [124], hairy
mouse [125], and hairless rat [126]. This last study, which compared in
vitro and in vivo data, also highlighted the potential role played by the
vasculature — however, no studies have been able to assess the perme-
ation of drugs between hair follicles and their surrounding vascular
network. Measurements of the effect of the vasodilator benzyl nicotinate
(BN) after topical application to the human forehead (rich in follicles),
forearm and calf (poor in follicles) suggest a significant role played by
the heavily vascularized of the hair follicles [127].
Studies comparing the penetration of a range of drugs through
human scalp and through abdominal skin demonstrate the influence
of greater follicle density [128]. Microscopy techniques suggest that
the infundibulum wall remains a significant barrier, albeit weaker
than the SC [66,122]. Visual evidence also suggests that permeation
beyond the infundibulum is favorable through the junction separat-
ing the outer and inner root sheaths [128,129]. Lademann et al.
conclude that massage induces a hair-shaft “pumping” mechanism
that improves penetration by pushing nanoparticles deeper into folli-
cles [130]. The same group has also found a positive correlation
between follicular penetration and the ‘activity’ of hair follicles
(measured in terms of sebum production and follicle growth) [131].
These results suggest that sebum flow does not impede permeation
through the follicular pathway [130], although the effects on drug
permeation are not clear. The influence of binding on permeation
has also been observed, particularly binding of lipophilic compounds
to the hair shaft cuticle [122,123]. Cone suggests that protein and
melanin are primarily responsible for hair-shaft binding and seques-
tration [132]. Reservoir effects within the cuticle [122,123] and sebaceous
gland [126,133] have been suggested, although further confirmation is
required.
Permeation through sweat gland epithelium appears to depend
significantly on lipophilicity, as is commonly observed in other epi-
thelia [133]. Jackson and Davis developed a 1D diffusion model for
permeation through the sweat duct, approximating it as a uniform
tube [134], and demonstrating that the sweat duct may act as a reser-
voir under occlusion and non-sweating conditions.
Lademann and co-workers have also visualized permeation into
sweat glands [135,136], with similar notions of ‘activity’ (albeit with
less well established criteria). The novel potential for delivery from
the eccrine sweat gland to the skin surface in human patients has
also been observed for intravenously (IV) administered compounds
[137,138]. Tight-junction-associated occludin, claudin 1 and claudin
4 are absent from those parts of the apocrine and apoeccrine ducts
positioned within the epidermis, offering a potentially more perme-
able route into the epidermis than the SC [119,139,140]. The lack of
a cornified cell layer further suggests the possibility of drug transport
via these glands, but this application remains to be investigated.
4. Global perspective
4.1. Overall distribution models
The aforementioned models, based on the mechanisms deduced
from experimental data and related skin physiology, describe penetra-
tion and distribution processes of a specific nature and at a specific
site. Many are developed to explain experimental data that is intention-
ally of a similarly specific nature, in order to improve understanding of
a particular phenomenon that would not be possible were other poten-
tially confounding factors not avoided. As a consequence of this
reductionist approach, global penetration and distribution models do
not generally comprise a combination of these state-of-the-art models.
Rather, information garnered from these models is typically fed into
global models of a relatively simple nature, typically involving linear
kinetics (i.e. compartment models). This raises the prospect that the
behavior exhibited by the more sophisticated model might not be avail-
able in the approximate global version: however, if the more sophisti-
cated model is linear, the underlying approximation can be improved
through the addition of further compartments.
At its simplest, one can then consider models such as those originally
proposed by Scheuplein and Blank [13] that represent the various
permeation pathways as a passage of diffusive resistances, connected
 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168 161

Fig. 5. (a) Schematic of drug distribution for topical (left) and intravenous (IV) (right) administrations: distribution is measured as local amounts A; also represented are the frac-
tions of administered drug available at the site of action (fa), and lost to metabolism in the skin (fm, skin) and in other organs (fm, body). (b) Total amount of butylparaben (●) recov-
ered as intact drug (♦) and as metabolized p-hydroxybenzoic acid (×) after application to excised rat skin as a 20% (w/v) drug suspension in phosphate-buffered saline [174].
Panel a: adapted from Ref. [141].
162 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168

in series (e.g. consecutive skin layers) or in parallel (e.g. SC route vs.

appendageal route), leading to expressions such as

!−1
1 1 1
kp;skin ¼ þ þ þ kp;appendages ð1:11Þ
kp;SC kp;VE kp;dermis

where the various local permeabilities are determined by factors out-

lined in the previous section.
Clearance from the skin also plays an important part in these
models [32,141]. For systemic delivery, the steady-state concentra-
tion of drug in plasma cSS can be approximated by Eq. (1.7), which
results from a mass balance of drug entering the skin (and therefore
the circulation, for a steady state) with that being cleared from the
circulation. Comparison with IV administration can provide informa-
tion about first-pass drug availability from topical application, once
the steady-state flux (or equivalent) has been established. This can
be achieved either by comparing the total AUC for topical and IV ad-
Fig. 6. Demonstration of ‘flip–flop’ kinetics, comparing excretion rates following oral
ministrations [142]: (open circles) and topic (closed circles) norephedrine HCl administrations.
Adapted from [175], cited in [3].
AUC topical DoseIV
Favailable ¼ ð1:12Þ
Dosetopical AUC IV
The kinetics of topical delivery into the systemic circulation can be
or by comparing excretion data for topical and IV administrations distinctive (in comparison with IV administration) due to the signifi-
[143] cant differences in absorption rates. Absorption is effectively instanta-
neous for IV administration, and therefore much faster than
Atopical DoseIV elimination (i.e. clearance): for topical delivery, absorption can be
Favailable ¼ ð1:13Þ
Dosetopical AIV much slower than elimination, leading to so-called ‘flip–flop kinetics’
where the usual rate-dependence on clearance observed from IV ad-
where A represents the amount excreted (from topical or IV adminis- ministration is ‘flipped’, and the absorption rate becomes the rate de-
tration). One can see the origin of these equations by considering the termining step (Fig. 6) [151,152].
various contributions to the first-pass skin availability, which takes into
account the combined effect of the fraction of topically applied drug fa
absorbed into the skin, as well as the fraction of drug 1 − fm, skin that re-
mains unaffected by skin metabolism. The amount excreted from the
urine is a fraction 1 − fm, skin of drug that remains unaffected by metab-
olism in the circulation, for both the IV and topical administrations,
which gives us (Fig. 5)
  
  fa 1−f m;skin 1−f m;body Atopical DoseIV
Favailable ¼ fa 1−f m;skin ¼   ¼ :
1−f m;body Dosetopical AIV

ð1:14Þ

Eq. (1.12) has been used to determine a skin first-pass availability

for topically administered fentanyl of 0.92 ± 0.33 [142]. Eq. (1.13)
has been used to determine the skin first-pass availability for
various commercial formulations of methyl salicylate, which varied
from 0.12 to 0.2 [143]; estradiol (Favailable = 0.075) [144]; diclofe-
nac (Favailable = 0.066) [145]; and nitroglycerin (Favailable = 0.72 ±
0.04) [144]. The various factors that affect first-pass availability, in-
cluding skin vascularization, metabolism and the availability of ap-
pendages [90], vary from site to site on the body, and experimental
data obtained for methyl salicylate [143] and nicotine [146]
demonstrate that first-pass availability is indeed site-dependent.
While our understanding of those enzymes responsible for transder-
mal drug metabolism continues to improve [147–149], the influence
of these enzymes on the kinetics of metabolized drugs is difficult to
determine. Specific enzymes that affect transdermally delivered drugs
include esterases (found in the viable skin) that play a role in the
first-pass metabolism of drugs such as methyl salicylate and nitroglyc-
erin [148]. The effect of esterases has been further investigated by Fig. 7. Schematic overview relating the effective clearance Cleff* from the site of action
during topical administration to the clearance from skin into the circulation Clskin, re-
skin pretreatment with an esterase inhibitor, and modeled with a turn clearance into the skin from the circulation Clr, and clearance from the circulation
two-layer diffusion model that predicted the decreasing lag time with ClIV.
increasing enzyme metabolic rate [150]. Adapted from Ref. [141].
 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168 163

Fig. 8. Prodrug, soft drug and pro-soft drug mechanisms of action. (a) Metabolism may strongly reduce the internal flux of the active compound, Jact,int, compared to the active com-
pound's external flux Jact,ext. (b) The prodrug approach consists in attaching a promoiety to the active drug which renders it inactive but increases its bioavailability, or external flux
Jact,ext. Cutaneous metabolism is harnessed to render the drug active. The resulting internal flux to the site of action, Jact,int, is greater than in the case of an unchanged drug (Fskin
designates the fraction of unchanged drug in skin). (c) A soft drug is the opposite of a prodrug. Skin metabolism is utilized to render an active drug inactive, which is eliminated
upon pharmacological action. (d) In the pro-soft drug approach, an inactive drug, with external flux Jinact,ext, is metabolized to an active compound which diffuses to the site of action
with internal flux Jact,int (Fskin designates the fraction of un-metabolized inactive drug). Following pharmacological action, the active drug is further metabolized to an inactive com-
pound and eliminated.
Figure adapted from Ref [141].

For cutaneous delivery, the clearance is now elimination (and poten-
tially metabolism) from the site of action, but the potential for drug to re-
turn to the site of action after uptake into the circulation must be taken
into account (Fig. 7): mass balance gives us cbClIV =cskinClskin −cbClr,
which combined with the definition of the effective skin clearance Cleff * the effective skin clearance is effectively just given by Clskin; however,
(Fig. 7) yields
 cb Cl Cl
Cleff ¼ ClIV ¼ IV skin
cskin ClIV þ Clr
where the Cl represent clearance from skin into the circulation (Clskin), re-
turn clearance from circulation into the skin (Clr), or clearance from the
circulation (ClIV), and cb and cskin represent concentrations in the blood
and skin, respectively. Binding may have an important influence on the
local therapeutically active concentration, as well as on clearance from
the site (Fig. 5). From Eq. (1.15) we identify two limiting cases: when
clearance from the circulation is much greater than the return clearance,
when the return clearance dominates, the effective clearance is much
* b b Clskin. In this second case, the drug exchange between
smaller, Cleff
skin and circulation is much greater than delivery from the skin surface.
ð1:15Þ
4.2. Prodrugs
While drug metabolism can reduce drug availability at the site of ac-
tion, metabolism can sometimes play a useful role in the improvement

Fig. 9. Comparison of in vitro vs. in vivo skin penetration data obtained from experiments with full-thickness human skin. The chemicals investigated are (a) nitrobenzene and (b)
2,4-dinitrochlorobenzene from [161]; (c) caffeine in water gel, (d) caffeine in petrolatum, (e) caffeine in ethylene glycol gel, (f) caffeine in benzoic acid and (g) testosterone from
[162]; (h) isofenphos [163]; (i) chloroform [164]; (j) lindane [165]; (k) chlorpyrifos [166,167]; (l) ortho-phenylphenol [168]; (m) methylene bis-benzotriazoyl tetramethylbutyl-
phenol (MBBT) and (n) titanium dioxide from [169].
164 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168

of topical drug delivery, or in the reduction of undesirable effects of a
drug beyond the site of action. Some of the difficulties in delivering
drug to the site of action, including low drug permeability through the
skin, fast drug elimination, or difficulties in formulation [153] can be
obviated through the addition of a promoiety to the original drug, form-
ing a prodrug. If the prodrug is metabolized to produce the desired drug
at the site of action, this approach can be used to deliver drugs more ef-
fectively. Metabolism can also play an important role in physiologically
deactivating drugs to non-toxic by-products once the desired pharma-
cological effect has been achieved at the site of action: drugs where
this behavior is observed are called soft drugs. It is also possible to
find a soft drug behavior in prodrugs: such compounds are known as
pro-soft drugs. The mechanisms of these approaches are outlined in
Fig. 8.
One of the key advantages of prodrugs for topical drug delivery is in
circumventing the skin barrier: a naltrexone prodrug improves skin
permeability 8-fold [154], while mouse and rat models demonstrate
improvements of 9-fold for 5-fluorouracil [155], 12-fold for ketorolac
[156] and almost 40-fold for nalbuphine [157]. The topical application
of the retinoid prodrug tazarotene may produce superior delivery to
orally administered alternatives due to esterase metabolism in the skin
that generates the more water-soluble active metabolite tazarotenic
acid [145,158].
systemic effect is the potency of a drug. Davis [170] has pointed
out that potency and skin penetration are equally important in deter-
mining the efficacy of topical products. He comments that direct skin
penetration of non-steroidal anti-inflammatory drugs is associated
with local efficacy without systemic adverse effects. He goes on to
point out that the topical application of other drugs, such as corticoste-
roids, retinoids and antihistamines is associated with systemic toxicity,
especially in severe skin disease.
In practice, therefore, the appropriate dosing of a topical product
must first begin with drug selection and, here, efficacy is a function
of both how much has penetrated and how potent the compound is.
One of the first authors to combine these concepts in a formal manner
was Lippold [171,172]. He defined the efficacy coefficient as the
ratio of the maximum flux divided by the drug potency dose. Thus,
greater efficacy is provided by either a higher flux or a more potent
(lower dose for the same effect) drug. Fig. 10 shows an illustration
of this approach using anti-inflammatory data from Cordero et al.
[173]. Here, it is evident that optimal skin penetration occurs at a
log octanol–water partition coefficient (log P) of 2 to 3 and that the

4.3. In vivo vs. in vitro results

Much of the available drug penetration data has been obtained

through in vitro experiments on excised skin, so the correlation be-
tween the values obtained in these results and the values that
would be expected in vivo is an issue of some significance. There is
an obvious importance in maintaining the skin viability through var-
ious means [159,160], but beyond this issue lies the question of how
well the essential conditions responsible for in vivo penetration are
reproduced in vitro. A number of studies [161–169] have performed
both in vitro and in vivo penetration studies, for the purposes of ex-
plicit comparison, by comparing the percentage of applied dose
absorbed in the receptor of the in vitro experiment to the percentage
of applied dose absorbed and excreted in the urine for the in vivo
experiment.
The overall picture is of a good correlation between in vitro and in
vivo data (Fig. 9), although a number of results from these studies
[161,162,168] show that the in vitro studies under-predict the in
vivo penetration. Such results might be explained by the lower tem-
perature of exposed skin during in vitro experiments than of skin
under clothing in vivo [162], or that a lag in excretion
underestimates the actual amount of penetrated drug in vivo [168].
Indeed, Cnubben et al. found a similar result in viable full-thickness
rat skin [168] — the more significant discrepancy for the same com-
pound (ortho-phenylphenol) in these rat studies was attributed to
greater compound metabolism in viable rat skin. Metabolism may also
play a role in the discrepancy for 2,4-dinitrochlorobenzene [161] and
testosterone [162].
One important exception to this observation, leading to the over-
prediction of the in vivo penetration from in vitro studies, is in the
case where the SC reservoir likely has an important influence in
vivo [165–167]. Important factors that influence in vivo penetration
for such drugs, including the effects of occlusion [165], or significant
partitioning into subcutaneous fatty tissue [166,167], are not neces-
sarily captured by the in vitro experiment. Denaturing of the skin
from the polar receptor fluid may also have influenced these results
[167].

4.4. Clinical response

Fig. 10. Using (a) maximal skin flux and (b) in vitro anti-inflammatory activity to esti-
mate (c) likely clinical efficacy of various non-steroidal anti-inflammatory drugs with
Percutaneous penetration does not necessarily guarantee response. varying values for logarithm octanol–water partition coefficient (log P).
Indeed, a key determinant in the choice of a drug for either local or Data from [173] and Scifinder data base.
 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168 165

more lipophilic drugs had greater in-vitro-based indices of anti- Appendix A. Summary of equations
inflammatory activity. As a consequence, diclofenac with log P > 4 has
the highest efficacy. As Davis [170] points out, the validity of this ap- Please refer to the text surrounding each equation for a full expla-
proach is evident from recent clinical studies showing the efficacy of nation of the relevant variables.
topical diclofenac and ketoprofen.
(1.1) SC homogeneous membrane model for solute concentration
(Fick's 2nd law):
5. Conclusions
∂C ðx; t Þ ∂2 C ðx; t Þ
¼ DSC :
∂t ∂x2
The complex structure of the skin reflects the myriad functions
that it performs, often (but by no means solely) related to its role as
(1.2) Usual boundary conditions for transdermal solute application:
a barrier separating us from the world outside. The variety of these
structures and functions makes it exceedingly difficult to give com- C ðx; 0Þ ¼ 0; C ð0; t Þ ¼ K SC C d ; C ðhSC ; t Þ ¼ 0 :
prehensive models that describe the penetration and distribution of
drugs passing through the skin. After many decades of investigation, (1.3) Amount of solute permeating the SC as a function of time:
and despite much progress on many of the fundamental questions,
!!
many details of transdermal drug transport remain elusive. However, DSC 1 2X ∞
ð−1Þn D 2 2
as experimental techniques – non-invasive imaging techniques in Q ðt Þ¼ K SC AC d hSC t− − exp −t 2SC π n :
2
hSC 6 π n¼1 n2 hSC
particular – continue to provide greater access to the details for trans-
dermal transport, our capacity to refine our models in light of these
(1.4) Long-time limit amount of solute permeating the SC as a func-
features continues to improve.
tion of time:
The overall picture of transdermal drug transport is one where the
SC provides the main penetration barrier for most compounds (for !
K D AC h2  
those drugs that readily enter the SC, the aqueous viable skin beneath Q ðt Þ ¼ SC SC d t− SC ¼ kp AC d t−t lag :
hSC 6DSC
provides the main resistance). Permeation through the SC is largely by
passive diffusion, although the nature of that process (transcellular vs
intercellular, the degree of corneocyte uptake) remains to be fully (1.5) Relationships defining permeability coefficient and lag time:
understood. The viable layers beneath the SC provide a less physically
constrictive environment for passive diffusion; however, there are var- K SC DSC h2
kp ¼ and t lag ¼ SC :
ious other processes that become relevant to the overall transport. hSC 6DSC
These include drug binding and sequestration, active transport, metab-
olism and (below the epidermal–dermal junction) transport mediated (1.6) Combining rule for permeabilities in series
by the vasculature and lymphatics. This last contribution can manifest
1 1 1
itself as a dispersive influence on the transport, as well as facilitating ¼ þ :
drug clearance. The appendages also provide an alternative pathway kp;epi kp;SC kp;VE
for topically administered drug — however, the capacity for drug to con-
tinue beyond the appendages into the surrounding skin or systemic cir-
culation is not entirely understood. (1.7) Steady-state solute concentration in viable epidermis, balan-
Because of the many different influences on overall drug transport cing steady-state delivery and clearance:
through skin, studies typically focus on improving our understanding
of one influence, restricting as much as possible the potential for con- kp;SC cV Favailable A J SS Favailable A
cSS ¼ ¼ :
founding effects. In order to provide a more global model of transdermal Cl Cl
transport, the information gained from these individually studies is
usually incorporated into standard pharmacokinetic models (usually (1.8) Steady-state solute concentration in viable epidermis, balancing
physiologically-based compartment models) that allow a representa- steady-state delivery and clearance, incorporating epidermal
tion of the drug distribution in relevant tissues. As our understanding drug binding/sequestration:
of skin transport improves to the extent that we may confidently juxta-
pose the more sophisticated local models, these pharmacokinetic kp;SC cV Favailable A
cSS ¼
models will also be able to provide a more sophisticated global picture Cl þ kp;SC Favailable A
of drug kinetics following topical administration.
An allied challenge is that of predicting transport properties. One mo- (1.9) Dermal diffusion–elimination model (elimination varying with
tivation for physiologically-based models is that the model parameters depth):
have a physiological interpretation. This in turn allows the possibility
that these parameters could be predicted from a fundamental under-
∂cðd; t Þ ∂2 cðd; t Þ
standing of the processes and interactions involved. Identifying those ¼ DðdÞ −kd P ðdÞcðd; t Þ:
∂t ∂2 x
physicochemical properties responsible for the various transport mecha-
nisms, and isolating their dependencies qualitatively and quantitatively, (1.10) Krestos et al. binding factor (accounting for dermal heterogeneity):
remain an on-going challenge. However, such predictive power is of fun-
damental importance in the continuing development of applications for ϕa
therapeutic topical drug administration. Binding factor ¼ ð1−ϕa Þ þ þ φlip fnon K P :
fu

(1.11) Combining rule for permeabilities in series and in parallel:

Acknowledgment
!−1
We are grateful to Ms. Klintean Wunnapuk for the preparation of 1 1 1
kp;skin ¼ þ þ þ kp;appendages :
Figs. 2, 5 and 7. kp;SC kp;VE kp;dermis
166 O.G. Jepps et al. / Advanced Drug Delivery Reviews 65 (2013) 152–168
(1.12) Drug availability obtained comparing AUC for topical and IV
administrations:
AUC topical DoseIV
Favailable ¼ : skin: theory and in vitro experimental measurement, AICHE J. 21 (1975) 985–996.
Dosetopical AUC IV
(1.13) Drug availability obtained comparing excretion from topical
and IV administrations:
Atopical DoseIV
Favailable ¼ : [23] C.A. Squier, The permeability of keratinized and nonkeratinized oral epithelium
Dosetopical AIV
(1.14) Drug availability, related to fraction of drug absorbed into the
skin and fraction of drug undergoing metabolism in skin
   [26] H.E. Bodde, I. van den Brink, H.K. Koerten, F.H.N. de Haan, Visualization of in
  f a 1−f m;skin 1−f m;body
Favailable ¼ f a 1−f m;skin ¼  
1−f m;body
Atopical DoseIV
¼ :
Dosetopical AIV
(1.15) Effective clearance, accounting for potential return of drug to
the site of action:
 cb Cl Cl
Cleff ¼ ClIV ¼ IV skin :
cskin ClIV þ Clr
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