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Drug Delivery
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Transdermal Delivery of Nicardipine: An Approach

to In Vitro Permeation Enhancement
a b b
Reza Aboofazeli , Hossein Zia & Thomas E. Needham
a
School of Pharmacy, Shaheed Beheshti University of Medical Sciences, Tehran, Iran
b
Department of Applied Pharmaceutics, College of Pharmacy, University of Rhode
Island, Kingston, Rhode Island, USA
Published online: 04 May 2015.

To cite this article: Reza Aboofazeli, Hossein Zia & Thomas E. Needham (2002) Transdermal Delivery of Nicardipine:
An Approach to In Vitro Permeation Enhancement, Drug Delivery, 9:4, 239-247

To link to this article: http://dx.doi.org/10.1080/10717540260397855

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 Drug Delivery, 9:239 – 247, 2002
Copyright °c 2002 Taylor & Francis
1071-7544 /02 $12.00 + .00
DOI: 10.1080 /1522795029009783 1
Transdermal Delivery of Nicardipine: An Approach
to In Vitro Permeation Enhancement
Reza Aboofazeli
School of Pharmacy, Shaheed Beheshti University of Medical Sciences, Tehran, Iran
Hossein Zia and Thomas E. Needham
Department of Applied Pharmaceutics, College of Pharmacy, University of Rhode Island, Kingston,
Rhode Island, USA
Downloaded by [Osaka University] at 22:10 15 July 2015
Nicardipine hydrochloride (NC-HCl), a calcium channel blocker
indicated for the treatment of chronic stable angina and hyper-
tension, seems to be a potential therapeutic transdermal system
candidate, mainly due to its low dose, short half-life, and high
Ž rst-pass metabolism. The objective of the present study was to
evaluate its  ux and elucidate mechanistic effects of formulation
components on transdermal permeation of the drug through the
skin. Solubility of NC-HCl in different solvent systems was deter-
mined using a validated HPLC method. The solubility of drug in
various solvent systems was found to be in decreasing order as
propylene glycol (PG)/oleic acid (OA)/dimethyl isosorbide (DMI)
(80:10:10 v/v) > PG > PG/OA (90:10 v/v) > polyethylene glycol
300 > ethanol/PG (70:30 w/w) > transcutol > dimethyl isosorbide
(DMI) > ethanol > water and buffer 4.7 > 2-propanol. Propylene
glycol was then selected as the main vehicle in the development of a
transdermal product. As a preliminary step to develop a transder-
mal delivery system, vehicle effect on the percutaneous absorption
of NC-HCl was determined using the excised skin of a hairless
guinea pig. Vehicles investigated included pure solvents alone and
their selected blends, chosen based on the solubility results. In vitro
permeation data were collected at 37±C, using Franz diffusion cells.
The skin permeation was then evaluated by measuring the steady
state permeation rate ( ux) of NC-HCl, lag time, and the perme-
ability constant. The results showed that no individual solvent was
capable of promoting NC-HCl penetration. Permeation proŽ les of
the drug through hairless guinea pig skin using saturated solutions
of drug were constructed. Among the systems studied, the ternary
mixture of PG/OA/DMI and binary mixture of PG/OA showed ex-
cellent  ux. The  ux value of the ternary system was nearly three
times higher than the corresponding values obtained for the binary
Received 29 January 2002; accepted 28 March 2002.
The authors would like to acknowledge the support by a grant from
Lohmann Therapie-Systeme GmbH & Co. and the Ministry of Health,
Treatment and Medical Education of I.R. of Iran for granting a sabbat-
ical leave.
Address correspondence to Hossein Zia, Dept. of applied Pharma-
ceutics, College of Pharmacy, Univ. of Rhode Island, Kingston, RI
02881, USA. E-mail: Hzia@uri.edu
239
solvent. A similar trend also was observed for the permeation con-
stant, while the values of lag time were reversed. The ternary mix-
ture was then selected as a potential absorption enhancement vehi-
cle for the transdermal delivery of drug. In general, higher  uxes
were observed through hairless guinea pig skin as compared with
the human stratum corneum. Based on the results obtained from
the release study of NC-HCl from saturated solutions of the drug,
a novel lecithin organogel (microemulsion-based gel) composed of
soybean lecithin, propylene glycol, oleic acid, dimethyl isosorbide,
and isopropyl myristate was developed as a possible matrix for
transdermal delivery of NC-HCl. In vitro percutaneous penetration
studies from this newly developed gel system through giunea pig
skin and human stratum corneum revealed that the organogel sys-
tem has skin-enhancing potential and could be a promising matrix
for the transdermal delivery of nicardipine. Furthermore, higher
permeation rates were observed when nicardipine free base was
incorporated into the gel matrix instead of hydrochloride salt.
Keywords Cosolvent System, Lecithin Organogel, Microemulsion-
Based Gel, Nicardipine Free Base, Nicardipine Hydro-
chloride, Penetration Enhancement, Transdermal Drug
Delivery
The transdermal route of administration has been recognized
as one of the potential routes for both the local and systemic
delivery of drugs. Transdermal delivery for systemic activity
provides several advantages, especially when drugs have low
oral bioavailability due to the extensive Ž rst passage metabolism
and/or a short half-life or even when drugs indicate some ad-
verse side effects following the oral administration. On the other
hand, the main limitation of transdermal delivery of drugs is
that the skin layers provide high resistance to the penetrant
molecules. Among these layers, the stratum corneum possesses
the most important barrier function, which results in the rate-
limiting step in percutaneous absorption. Consequently, it is
difŽ cult to deliver many drugs at rates sufŽ cient to provide clin-
ically effective plasma concentrations. Different strategies have
 240 R. ABOOFAZELI ET AL.
been developed to minimize the skin’s barrier function, such as
employing appropriate components with skin-enhancing poten-
tial (Sinha and Kaur 2000; Squillante et al. 1998, 2000; Singh
et al. 1996) or applying electricity and ultrasound, referred to as
iontophoresis and sonophoresis techniques, respectively (Curdy
et al. 2001; Guy 1998; Mitragorti and Kost 2000; Mitragorti et al.
1995).
Nicardipine hydrochloride (NC-HCl), a dihydropyridin e cal-
cium channel blocker, is indicated in the management of hyper-
tension and angina pectoris and related cardiovascular disorders.
Although this drug is rapidly absorbed from the gastrointestinal
tract following oral administration, its extensive Ž rst-pass meta-
bolism, along with its short half-life (Sorkin and Clissold 1987;
Frishman 1989) make it a potential candidate for transdermal de-
livery. A transdermal delivery system also would have the advan-
tage of constant delivery of drug into the blood circulation, which
in turn would decrease the interindividual variability in plasma
level due to the differences in bioavailability and metabolism
Downloaded by [Osaka University] at 22:10 15 July 2015
following oral administration. However, with an initial dose of
60 mg per day and an oral bioavailability of 35%, a transder-
mal device with a 20-cm2 release surface area should deliver
NC-HCl at rates in excess of 43.75 ¹g/cm¡2 ¢hr¡1 . Nicardipine
hydrochloride itself has poor permeability in skin transport and
hence is not capable of penetrating through the skin at a rate
necessary to provide a therapeutic concentration.
Very few reports have been published investigating the in u-
ence of vehicles on the enhancement of percutaneous absorption
of NC-HCl and nicardipine free base (NC-base). These works
are limited to the reports of Seki et al. (1987, 1989), Yu et al.
(1988), and Diez et al. (1991). Thus, the three principal objec-
tives of the present study were: 1. To examine the comparative
effect of several pure or mixed solvent systems on the  ux of
NC-HCl through hairless guinea pig skin. 2. To perform a com-
parative study of percutaneous absorption of the drug dissolved
in the selected solvent systems. 3. To optimize a transdermal
formulation, with respect to in vitro human skin  ux by devel-
oping a semisolid preparation in an attempt to obtain a semisolid
reservoir transdermal device with sufŽ cient efŽ cacy.
To achieve the Ž nal objective, it seemed more reasonable to
look for a carrier that interacts with the human skin and allows
the permeation of the drug into the skin. In this regard, lecithin
organogels (so-called microemulsion-based gels) were consid-
ered as potentially pharmaceutically acceptable carriers for the
transdermal transport of nicardipine. These thermodynamically
stable, transparent organogels are interesting for their high sol-
ubilizing capacity for various molecules and may be used as a
matrix for the transdermal delivery of drugs with relatively great
in ux (Dreher et al. 1996, 1997).
MATERIALS
Nicardipine hydrochloride (NC-HCl), cis-oleic acid (OA),
and olive oil were supplied by Sigma (St. Louis, MO, USA).
Acetonitrile, methanol, monobasic potassium phosphate, 2-
propanol, and propylene glycol (PG) were purchased from Fisher
ScientiŽ c (Fair Lawn, NJ, USA). Isopropyl myristate and al-
mond oil were obtained from Ruger Chemical Co. (Irvington,
NJ, USA). Polyethylene glycol 300, dimethyl isosorbide (DMI),
and triacetin were provided by Union Carbide Chemicals
(Danbury, CT, USA), ICI (Wilmington, DE, USA), and Eastman
Chemical Co. (Kingsport, TX, USA), respectively. Transcutol
was a gift from Gattefosse’ (CedexFrance, France) and ethanol
was purchased from AAPER Alcohol and Chemical Co.
(Shelbyville, KY, USA). Nicardipine free base (NC-base) was
prepared from the hydrochloride salt via an acid-base reaction
with NaHCO3 and recovery by chloroform. Soybean lecithin
(Epikuron 200, with 95% w/w phosphatidylcholine) was pro-
vided by Lucas Meyer Company (Germany) and was used with-
out further puriŽ cation. All reagents were used as obtained.
METHODS
High Performance Liquid Chromatography Assay
A Waters liquid chromatograph equipped with a SB-CN
Zorbax column (Agilent Technologies, Palo Alto, CA, USA),
two Waters 515 HPLC pumps, a Waters 746 data module, a Wa-
ters 717 plus autosampler, and a Waters Lambda-Max model
480 LC spectrophotometer (set at 240 nm) were used for analy-
sis. The samples were eluted using acetonitrile/methanol/0.02 M
monobasic potassium phosphate solution with a ratio of
20:30:50 v/v at 37± C and a  ow rate of 1 mL/min. Calibra-
tion was carried out by 10 runs of NC-HCl standard curves on
different days, and a composite standard curve with a correlation
coefŽ cient of r2 D 0.9999 was then obtained over the concen-
tration range of 1–100 ¹g/ml.
Solubility Determinations
The solubility of the drug in a variety of media was deter-
mined, in triplicate, at room temperature. An excess amount of
drug was suspended in the medium in screw-capped vials and
equilibrated in a rotating bottle for 24 hr. Then, an aliquot of the
suspensions was transferred to a 1-mL microcentrifuge Ž lter, Ž t-
ted with a 0.22 ¹m nylon Ž lter (Corning Incorporated, Corning,
NY, USA), and centrifuged. The Ž ltrates were appropriately di-
luted with methanol before assaying by HPLC.
Preparation of Suspensions of NC-HCl
An excess of NC-HCl was added to the pure solvents and their
blends and the initial suspensions thus obtained were agitated for
24 hr at room temperature. These suspensions were employed
as donor formulations in the permeation studies.
Preparation of Nicardipine-Loaded Lecithin Organogels
The lecithin organogel containing drug was prepared at room
temperature by Ž rst dissolving nicardipine (either as a salt or
base) in the PG/OA/DMI ternary mixture, followed by the addi-
tion of isopropyl myristate and lecithin. The mixture was stirred
for sufŽ cient time until a clear relatively viscous solution was
 TRANSDERMAL DELIVERY OF NICARDIPINE
obtained. The gel was formed after dropwise addition of a spe-
ciŽ c amount of water.
Preparation of Stratum Corneum
The human skin samples were purchased from the National
Disease Research Interchange. Epidermal membranes were pre-
pared by the heat separation technique. Simply, the excess fatty
and connective tissues were removed from the skin and then
small pieces of circular sections with a 3-cm diameter were
punched out and immersed in a water bath at 60 § 1± C contain-
ing 0.002% (w/v) sodium azide, for about 60 sec. Using forceps,
the epidermis was teased away from the underlying dermis and
 oated on the surface of distilled water in a beaker, washed, then
transferred onto an aluminum foil, dried, and stored in a freezer
until use.
In Vitro Permeation Studies
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The passage of the solubilized drug (nicardipine hydrochlo-
ride) from suspensions, prepared with various pure solvents and
their blends, through the hairless guinea pig or human skins was
studied using  at  ange Franz diffusion cells. The active dif-
fusion area and the volume of the receiver compartment were
0.64 cm2 and 5.3 mL, respectively. The membrane (full thick-
ness hairless guinea pig skin or human stratum corneum) was
mounted carefully onto the diffusion cell with the stratum
corneum side facing the donor compartment. The donor and the
receiver compartments were clamped together and the receiver
compartment was then Ž lled with 0.01 M phosphate buffer, pH
4.7 solution and stirred with a magnetic stirrer. After reaching
37± C temperature, 300 ¹L of each suspension or 400 mg of
lecithin gel formulation of the drug was placed in the donor
compartment. At speciŽ c intervals, the receptor solution was
completely withdrawn and the acceptor compartment was re-
Ž lled with fresh buffer solution. The drug concentration in the
receiver  uid samples was assayed by HPLC.
Data Analysis
The in vitro skin  ux was determined from the Fick’s law of
diffusion written as:
1 dM DP(C d ¡ Cr )
Js D D [1]
S dt h ethanol/PG, and ternary system of propylene glycol/oleic acid/
where Js is the skin  ux (¹g¢cm¡2 ¢hr¡1 ); P, partition coefŽ cient
of drug between donor and membrane; dM=dt, the amount of
drug permeated per unit time (¹g¢hr¡1 ); S, the surface area (cm2 )
of diffusion; D, the drug diffusion coefŽ cient (cm2 ¢hr¡1 ); Cd
and Cr are the drug concentration in the donor and receptor
compartments (¹g¢cm¡3 ), respectively, and h is the thickness of
the membrane (cm).
If sink conditions hold in the receptor compartment, Cr ¼ 0,
then one can write that:
dM DPSCd
D D K p SCd [2]
dt h
241
in which
DP
Kp D
h
Where K p is permeability constant and has a unit of cm/s.
Integration of equation (2) between limits of zero and t where
M is zero at t D 0, yields a straight line equation as follows:
M D K p SCd t
or
M D K p SC d (t ¡ t L ) [3]
Where t L is the lag time.
Penetration curves were constructed by plotting the cumula-
tive amount of drug penetrated through unit area of the mem-
brane versus time. The steady state  ux was determined by re-
gression analysis of the linear portion of the plot. The permeation
study results are the average of six replicates. Permeability con-
stants were calculated by dividing the slope by the corresponding
concentration of the drug in the donor compartment, and the lag
time also was determined by extrapolating the linear portion of
the cumulative amount-time curve to the abscissa. The enhance-
ment ratio (ER) relative to pure solvent also was used to compare
drug  ux among some of the formulations studied.
Statistical Analysis
Results are reported as mean § SD. The permeation param-
eter,  ux, obtained for various vehicles, was compared using
one-way ANOVA. Differences between the treatments were as-
sumed to be signiŽ cant at p < .05. All data analysis and statistical
calculations were performed using Minitab Statistical Software
13.1 for Windows (PA, USA).
RESULTS
Solubility Measurements
Table 1 shows the solubility of NC-HCl in pure solvents,
binary systems of propylene glycol/oleic acid (PG/OA) and
dimethyl isosorbide (PG/OA/DMI) at room temperature. The
highest solubility (about 33 mg/mL) observed in the ternary
PG/OA/DMI (80:10:10 v/v) solvent system amounted to about
1.2 and 3.2 times greater than that in PG and DMI alone, re-
spectively. However, the solubility of NC-HCl in binary co-
solvent systems showed a decrease when oleic acid or ethanol
was added to the pure PG solvent. In general, the solubility re-
sults showed an decreasing order of PG/OA/DMI (80:10:10 v/v)
PG > PG/OA (90:10 v/v) > polyethylene glycol 300 > ethanol/
PG (70:30 w/w) > transcutol > DMI > ethanol > water and
buffer 4.7 > 2-propanol. Nicardipine hydrochloride is spar-
ingly soluble in normal saline, isotonic buffer (pH 4.5), triacetin,
 242 R. ABOOFAZELI ET AL.

TABLE 1
Saturation solubility of nicardipine hydrochloride in various
solvent and cosolvent systems

Solvent Solubility SD
Water 4.072 0.146
Normal saline 0.242 0.046
Buffer 7.4 Not measurable —
Olive oil 0.029 0.004
Almond oil 0.045 0.017
Propylene glycol (PG) 27.999 0.871
Polyethylene glycol 300 21.718 0.740
Ethanol/PG (70:30 wt%) 21.185 0.634
Ethanol 8.404 0.090
FIG. 2. Release proŽ le of nicardipine hydrochloride through hairless guinea
2-Propanol 1.149 0.095
pig skin, from PG/OA and PG/OA/DMI suspensions .
Dimethyl isosorbide (DMI) 10.220 0.568
Oleic acid (OA) Not measurable —
portion of the plots (cumulative amount permeated per unit area
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Triacetin 0.158 0.019

versus time), lag time, and permeability constants are shown in
Transcutol 13.624 2.454
Table 2. The results showed that no individual solvent was ca-
Isopropyl myristate Not measurable —
pable of promoting sufŽ cient NC-HCl permeation to meet our
Buffer 4.7 4.070 0.191
objectives. Although the solubility of NC-HCl in PG and PEG
Isotonic buffer 4.5 0.219 0.019
300 was higher than that in other pure solvents, there is no sub-
PG/OA (90:10 v/v%) 24.599 2.12
stantial increase in the permeation of drug from the PG and PEG
PG/OA/DMI (80:10:10 v/v%) 32.861 0.336
300 suspensions. The high  ux observed after 20 hr with the DMI
suspension may be due to skin damage. The one-way ANOVA
almond oil, and olive oil and is practically insoluble in isopropyl performed on the release data indicated no signiŽ cant difference
myristate, buffer (pH 7.4), and oleic acid due to its physicochem- in  ux and permeability constant, among these suspensions at
ical nature. ® D 0.05 ( p D 0.186).
Figure 2 compares the enhancement potential of the binary
and ternary cosolvent formulations, prepared by blending PG/OA
Release ProŽle Through Hairless Guinea Pig Skin (90:10 v/v%, f 5 ) and PG/OA/DMI (80:10:10 v/v%, f 6 ), re-
from Saturated Solutions spectively. Table 2 summarizes the permeation parameters of
Based on the results of the solubility studies, PG, PEG 300, NC-HCl from these cosolvent systems through guinea pig skin.
DMI, and transcutol were chosen for percutaneous absorption Table 3 displays the corresponding enhancement ratios. In gen-
testing. Figure 1 shows the permeation proŽ les of NC-HCl from eral, the addition of 10% oleic acid and 20% OA/DMI mixture
suspensions (saturated solutions) through hairless guinea pig (with the ratio of 1:1, v/v) to the PG-based suspension promoted
skin. The corresponding NC-HCl  ux calculated from the linear  ux to a much greater degree with shorter lag times than the
corresponding pure solvents. Also, as shown in the Table 2, the
lowest coefŽ cients of variation for the permeation parameters
(Q24 , J, and K p ) were observed when the drug was suspended in
the binary and ternary mixtures, compared with those obtained
from pure single solvent systems. The decrease in the release rate
after 16 hr may be attributed to the absence of sink conditions,
due to the increase period of the sampling. SigniŽ cant differ-
ences were observed in  ux values from f 5 and f 6 formulations
compared with the pure solvents ( p < .0005).

Release ProŽle Through Hairless Guinea Pig Skin

FIG. 1. Release proŽ le of nicardipine hydrochloride through hairless guinea
pig skin, from suspensions prepared with various pure solvents.
from Lecithin Organogels
The lecithin-matrix gel containing nicardipine (as salt and
base) was Ž rst studied for permeability of the drug across the
hairless guinea pig skin. Table 4 indicates the composition of the
drug-loaded lecithin gel formulations investigated in this study.
 TRANSDERMAL DELIVERY OF NICARDIPINE 243

TABLE 2
Permeation parameters of nicardipine hydrochloride in pure base solvents, binary and ternary mixtures, through hairless
guinea pig skin (mean § SD)

Q24 a Jb Kpd tL e
2c
Vehicle Composition (¹g¢cm¡2 ) (¹g¢cm¡2 ¢hr¡1 ) r (cm¢hr¡1 ¢105 ) (hr)
f1 Propylene glycol 8.66 § 6.62 0.58 § 0.40 0.9735 2.07 § 1.44 10.10
(76.4%) (69.6%) (69.6%)
f2 Polyethylene glycol 400 6.68 § 4.96 0.47 § 0.33 0.9923 2.41 § 1.51 9.56
(74.3%) (70.2%) (62.6%)
f3 Dimethyl Isosorbide 72.73 § 88.09 1.39 § 1.13 0.9909 13.60 § 11.08 5.34
(121.1%) (81.5%) (81.5%)
f4 Transcutol 9.44 § 14.63 0.57 § 0.89 0.9876 4.22 § 6.57 8.03
(155.0%) (155.7%) (155.7%)
f5 Propylene glycol/oleic acid 2545.20 § 1024.70 f 115.15 § 39.05 0.9933 468.13 § 158.76 3.38
(40.3%) (33.9%) (33.9%)
f6 Propylene glycol/oleic acid/ 7361.35 § 2827.73 471.37 § 252.24 0.9917 1434.45 § 767.60 1.72
Downloaded by [Osaka University] at 22:10 15 July 2015

dimethyl isosorbide (38.4%) (53.5%) (53.5%)

a
Cumulative amount of drug permeated per unit area. b Flux. c Correlation coefŽ cient of curve Ž t. d Permeability constant. e Lag time obtained
from the mean release curve. f Q 26 .
Numbers in parentheses show the coefŽ cient of variation.

Figure 3 shows the total amount of nicardipine permeated into
the receiver after a 28-hr period when the donor is composed of
the different weight percentages of nicardipine (either as salt or
base) solubilized in lecithin organogel.
It should be noted that NC-base could be solubilized in the gel
matrix to a greater extend than NC-HCl. In this regard, although
the concentration of NC-base in formulation f8 is approximately
2-fold greater than the concentration of NC-HCl in formulation
f7 , it has been observed that the release rate obtained from the f8
gel formulation was about 16-fold higher than that obtained from
the f7 gel formulation. This difference in permeation rate may be
mainly attributed to the fact that the more lipophilic free base can
penetrate the skin much more readily than the hydrochloride salt
(Table 5). As can be seen in Figure 3, a higher amount of drug
was released when the drug concentration was raised 3.57% to
9.34% and 13.0% (w/w), although the increase in the cumulative
amount of drug permeated was not directly proportional to the
increase in drug content. Although the single-factor ANOVA
performed on the  ux values of the different gel formulations
containing NC-base ( f8 , f9 , f10 ) showed no signiŽ cant differ-
ence ( p D .081), the difference in the transport rate between
NC-HCl and NC-base loaded gels was found to be signiŽ cant
( p < .0005).
Release ProŽle Through Human Stratum Corneum
The cumulative amount-time curve proŽ les of NC-HCl and
NC-base through human stratum corneum from three vehicle

TABLE 3
Effect of binary and ternary cosolvent systems on the
permeation of nicardipine hydrochloride, through hairless
guinea pig skin

Propylene Oleic Dimethyl

glycol acid isosorbide
Vehicle (%) (%) (%) ERa
f5 90 10 — ERPG : 199, EROA : 45
f6 80 10 10 ERPG : 814, EROA :
182, ERDMI : 339
a
ERPG , EROA , and ERDMI designate the ratio of  ux of nicardipine
hydrochloride from mixtures relative to the  ux of the drug from pure
solvent. The average  ux of nicardipine hydrochloride from oleic acid FIG. 3. Release proŽ le of nicardipine (as salt and base) through hairless
saturated solution was found to be 2.59 ¹g/cm2 /hr. guinea pig skin, from lecithin organogel s composed of various drug contents.
 244 R. ABOOFAZELI ET AL.

TABLE 4
Composition of drug-loaded lecithin organogels

Drug PG/OA/DMIa IPM Lecithin Water

Formulation (%) (%) (%) (%) (%)
f7 NC-salt, 1.54 49.69 5.12 37.65 6.00
f8 NC-base, 3.57 44.85 8.78 36.60 6.20
f9 NC-base, 9.34 40.37 7.80 33.64 8.85
f 10 NC-base, 13.00 37.30 8.30 28.15 13.25
a
This mixture was prepared with the ratio of 80:10:10 by volume.

formulations are shown in Figure 4 and the corresponding per- DISCUSSION

meation parameters are summarized in Table 6. It should be
noted that the time to reach the steady state  ux of NC-base from
lecithin gel formulation ( f10 ) was remarkably shorter than that
for the other two vehicle formulations studied. The cumulative
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One of the major priorities in pharmaceutical research is to
design and develop formulations for nonoral administration in
an attempt to overcome potential disadvantages, such as Ž rst-
pass effect, low bioavailability, and the adverse effects often

amount of NC-base released from gel matrix was 2.1-fold higher
than that from tertiary solvent formulation with NC-base and
2.8-fold greater than tertiary solvent formulation contained NC-
HCl. This suggests that the gel matrix possesses more skin pen-
etration enhancement effect. The permeability coefŽ cients of f11
and f6 vehicles were found to be 32.18 £ 10¡5 and 13.75 £ 10¡5 ,
respectively, while the  ux were 10.62 ¹g¢cm¡2 ¢hr¡1 and
4.52 ¹g¢cm¡2 ¢hr¡1 , respectively.
The high  ux and other permeability parameters of NC-base
might be due to the increased lipophilicity of the drug as a free
base in the medium. This Ž nding is in good agreement with that
observed in the comparative release study of NC-HCl and NC-
base, dissolved in gel formulations, through guinea pig skin.
However, the sharp increase in the amount of NC-HCl perme-
ated after 20 hr may have been from the loss of skin integrity.
Statistical analysis showed a signiŽ cant difference in the  ux
values between the three formulations studied (p D .001).
associated with oral delivery. In this regard, the potential of skin
as a site of administration for systemically active drugs has been
realized. However, the skin, and especially the stratum corneum,
behaves as a barrier and often provides signiŽ cant resistance to
penetration. For this reason, candidate drugs for transdermal de-
livery require favorable physicochemical properties for the suc-
cess of percutaneous absorption. A desired transdermal delivery
system (TDS) should provide a short lag time and adequate  ux.
To achieve these goals, a candidate drug is required to have suf-
Ž cient solubility in the vehicle and a high permeability constant.
The pharmacokinetic characteristics of NC-HCl, a dihydropy-
ridine calcium antagonist commonly used for the treatment of
hypertension, make this drug a potentially suitable candidate for
formulation in a TDS (Diez et al. 1991). However, its low  ux, as
with  ux of other drugs in this category (Squillante et al. 2000)
necessitates a unique novel method to enhance its permeation
through the skin.

TABLE 5
Permeation parameters of nicardipine hydrochloride and nicardipine free base dissolved
in lecithin gels, through hairless guinea pig skin (mean § SD)

Q24 a Jb Kpd (§SD) tL e

2c
Formulation (¹g¢cm¡2 ) (¹g¢cm¡2 ¢hr¡1 ) r (cm¢hr¡1 ¢105 ) (hr)
f7 5.97 § 9.29 0.32 § 0.44 0.9896 2.12 § 2.96 3.9
(155.6%) (139.5%) (139.5%)
f8 118.64 § 39.48 6.26 § 2.00 0.9981 17.89 § 5.71 4.8
(33.3%) (31.9%) (31.9%)
f9 175.88 § 88.32 10.02 § 5.18 0.9972 10.73 § 5.54 6.3
(50.2%) (51.7%) (51.7%)
f 10 (n D 12) 213.55 § 111.16 12.29 § 5.91 0.9945 9.45 § 4.54 7.4
(52.1%) (48.1%) (48.1%)
a
Cumulative amount of drug permeated per unit area. b Flux. c Correlation coefŽ cient of curve Ž t.
d
Permeability constant. e Lag time obtained from the mean release curve.
Numbers in parentheses show the coefŽ cient of variation.
 TRANSDERMAL DELIVERY OF NICARDIPINE 245

PEG 300 (21.718 mg/mL), DMI (10.220 mg/mL), and transcutol

FIG. 4. Release proŽ le of nicardipine (as salt or base) through human stratum
corneum from a 13.0% lecithin organoge l and ternary mixture.
Results obtained from the solubility experiments showed that
NC-HCl has adequate solubility in PG, PEG-400, DMI, and
Downloaded by [Osaka University] at 22:10 15 July 2015
(13.624 mg/mL).
In other words, no individual solvent possessed the capabil-
ity of promoting NC-HCl permeation through guinea pig skin.
That is, despite of the difference between the drug-solubilizing
capacity of the solvents, it seems that the thermodynamic ac-
tivity of the drug in all the different suspensions is roughly the
same. This Ž nding suggests that the permeation rate of the drug
is independent of the nature of the pure solvents employed.
Squillante et al. (1998) have investigated the effect of pure
or mixed solvent systems on the skin permeation of nifedip-
ine through excised mouse skin. They have shown a positive
synergistic interaction among the formulation component PG,
OA, and DMI and noted that a mixture of these components
would yield optimal  ux and lag time. The present in vitro per-
meation experiments showed that a binary mixture of PG and
OA (90:10 v/v) and a ternary mixture of PG, OA, and DMI
(80:10:10 v/v) had a remarkably positive effect on the pen-

transcutol. Consequently, these pure solvents were examined
for their in uence on drug permeation. From the thermodynamic
point of view, the steady state  ux can be expressed by (Higuchi
1960):
ad D
J D
am h
where ad is the thermodynamic activity of the drug in the vehi-
cle, am is the activity coefŽ cient of the drug in the skin, D and h
are diffusion coefŽ cient of the drug in the skin and skin thick-
ness, respectively. This equation simply shows that the maxi-
mum  ux may be achieved when the drug is at maximum ther-
modynamic activity, provided that all the terms in the equation
remain constant. Therefore, for the release studies, suspensions
rather than solutions were used to guarantee a constant level
of activity. Although the solubility of NC-HCl in PG was the
highest among the pure solvents studied (27.999 mg/mL), as
depicted in Table 2, the in vitro skin permeation rate was vir-
tually the same for all the high solubility solvents, including
etration of NC-HCl. The results were even more satisfactory
with the application of the ternary system. It should be noted
that when OA was added to PG, the resulting binary system
so formed had lower solubility for NC-HCl (24.599 mg/mL),
but had the effect of increasing permeation. In other words, the
[4] binary mixture with comparatively lower solubility promoted
 ux 200-fold greater (115.15 ¹g¢cm¡2 ¢hr¡1 ) with a consider-
ably shorter lag time (3.38 hr) than the corresponding PG sus-
pension ( ux value of 0.58 ¹g¢cm¡2 ¢hr¡1 and lag time value of
10.10 hr).
Furthermore, it was found that with an increase of only
5 mg/ml in solubility in the ternary system over that in the
pure solvent PG, NC-HCl yielded a calculated  ux of 800 times
higher (471.37 ¹g¢cm¡2 ¢hr¡1 ). These results imply that the level
of the enhancement was signiŽ cantly dependent upon the choice
of the cosolvent vehicle rather than drug solubility. Since the
thermodynamic activity of NC-HCl in the different cosolvent
systems is approximately the same, the rapid skin permeation
would seem to be related to changes in the diffusion coefŽ cient

TABLE 6
Permeation parameters of nicardipine hydrochloride and nicardipine free base in ternary mixtures
and lecithin gel, through human stratum corneum (mean § SD, n D 12)

Q24 a Jb Kpd tL e
2c
Vehicle-drug (¹g¢cm¡2 ) (¹g¢cm¡2 ¢hr¡1 ) r (cm¢hr¡1 ¢105 ) (hr)
Gel (13.0%)/NC (base) 432.46 § 275.75 20.84 § 12.74 0.9982 16.03 § 9.80 3.3
( f 10 ) (63.8%) (61.1%) (61.1%)
PG-OA-DMI/NC (base) 201.21 § 151.66 10.62 § 8.56 0.9996 32.18 § 25.93 5.1
( f 11 ) (75.37%) (80.6%) (80.6%)
PG-OA-DMI/NC (salt) 153.82 § 87.70 f 4.52 § 4.28 0.9729 13.75 § 13.03 6.8
( f6) (57.0%) (94.8%) (94.8%)
a
Cumulative amount of drug permeated per unit area. b Flux. c Correlation coefŽ cient of curve Ž t. d Permeability
constant. e Lag time obtained from the mean release curve. f n D 8.
Numbers in parentheses show the coefŽ cient of variation.
 246 R. ABOOFAZELI ET AL.
of drug in the skin and/or the activity coefŽ cient of drug in the
skin, which are induced by the penetration of these solvents (Seki
et al. 1987). It has also been shown that both PG and DMI readily
permeate the skin, increasing the solubility of the penetrant in
the stratum corneum, and that OA facilitates penetration into the
skin by disrupting the  uidity of the stratum corneum (Squillante
et al. 1998; Potts et al. 1991). Therefore, it would seem that ve-
hicles that permeate the skin also promote NC-HCl  ux. Table 2
demonstrates that the coefŽ cients of variation calculated for the
permeation parameters were quite considerable for all solvents
and cosolvents, which could be attributed to the skin variabil-
ity. However, the least variation was observed when binary and
ternary solvent system were used for release study.
Although it is not possible to extrapolate the animal data
to humans, if we assume that the hairless guinea pig skin is
10 times more permeable than the human skin, one would ex-
pect that the ternary mixture could yet be the optimized liquid
vehicle for the transdermal delivery of NC-HCl at the desired
Downloaded by [Osaka University] at 22:10 15 July 2015
rate. However, when comparison was made of the intrinsic abil-
ity of the ternary mixture to promote drug permeation through
human stratum corneum, this mixture had little effect on the skin
permeability of the drug (the  ux value of 4.52 ¹g¢cm¡2 ¢hr¡1
was obtained). Thus, a decision was made to evaluate the re-
lease kinetics of NC-base through skin membrane. Figure 4
compares the amount of NC-base permeated into the receptor
chamber through human stratum corneum, when the free base
was dissolved in PG/OA/DMI mixture ( f 11 ) with the same con-
centration as formulation f 6 . Though the enhancement was not
substantial and the amount delivered still remained below the
therapeutic level, drug permeation was improved. This result
indicates that the more lipophilic free base penetrates the skin
more readily than the hydrochloride salt (Yu et al. 1998). It has
been proposed that PG and DMI displace the water in the stra-
tum corneum required to decrease the lipid rigidity and cause
dehydration (Squillante et al. 1998;). NC-base has a very poor
solubility in water (Kou et al. 1993), hence, one would expect
diffusion of the base through dehydrated skin likely would be
improved, due to this displacement of stratum corneum water.
In apolar organic solvents, soybean lecithin (a mixture of
phosphatidylcholine, phosphatidylinositol, and phosphatidy-
lethanolamine) can form a thermoreversible, isotropic, nonbire-
fringent gel-like system, so-called microemulsion-based gel or
organogel, characterized by considerably high viscosity and op-
tical transparency (Bhatnagar and Vyas 1994; Scartazzini and
Luisi 1998). Lecithin, a naturally occurring surfactant, is ca-
pable of forming reverse micelle-based microemulsions in an
apolar environment due to its geometrical constraints. It is be-
lieved that upon addition of a speciŽ c amount of water, the
small reverse micelles present initially in the lecithin-organic
solvent mixture tend to grow monodimentionally into long  ex-
ible and cylindrical giant micelles, above a critical concentration
of lecithin. These giant micelles then build a continuous entan-
gled network with a high macroscopic viscosity (Schurtenberger
et al. 1990). Lecithin organogels have attracted attention as a
biocompatible matrix for trandermal delivery of drugs. These
systems are particularly interesting (Dreher et al. 1996, 1997)
because:
² They are thermoreversible systems that become liquid
with lower viscosity at temperatures above 40± C and
regain high viscosity by cooling.
² They are capable of solubilizing various guest mole-
cules (lipophilic, hydrophilic, and amphoteric).
² They do not cause skin irritation.
² They possess long-term stability.
² They are transparent, allowing the use of spectroscopic
methods to detect possible structural changes of the
guest molecules.
The formation of lecithin organogels in the system lecithin/
organic solvent/water with various organic solvents, such as lin-
ear and cyclic alkanes, ester of fatty acids, and amines, have been
reported in the literature (Scartazzini and Luisi 1988). However,
for pharmaceutical and medical applications, the organogels
need to be composed of biocompatible components. This re-
search relates to a novel method of administration of a phar-
macologically active dihydropyridin e calcium channel blocker,
nicardipine, to promote the absorption and provide therapeu-
tically effective concentration in the bloodstream. The method
comprises the drug nicardipine base in combination with a
lecithin organogel carrier vehicle, composed of nontoxic ad-
juvants selected from among propylene glycol, dimethyl isosor-
bide, oleic acid, and isopropyl myristate that have no physiolog-
ical in uences in the body other than they increase the rate of
absorption across the body membranes.
In general, the results obtained from the release study through
guinea pig skin showed that the permeation rate from a ternary
mixture composed of PG/OA/DMI was signiŽ cantly greater than
that observed with the corresponding lecithin gels. The explana-
tion for this difference is that there are some factors, including
drug-micelle interaction (i.e., the location of the guest molecule
in the reverse micelles, either in hydrophilic core or in lipophilic
hydrocarbon chains), the amount of the entrapped drug inside
the micelle cores, and the quantity of water used as the gel in-
ducer (or viscosity modiŽ er) which strongly affect the transport
rate. The comparison between the trandermal transport curves
obtained from organogels composed of NC-HCl (1.54% w/w)
and NC-base (3.57, 9.34, and 13.0% w/w) through guinea pig
skin reveals two points: an increase in the drug content leads to
an increase in the  ux, and the transport rate with NC-base is
at least 20 times higher than NC-HCl. The latter reconŽ rms the
higher penetrability of the free base.
Although the apolar phase of the gel is composed of a mixture
of PG/OA/DMI with its own skin penetration enhancement po-
tential, it also has been suggested that lecithin gels may slightly
disorganize the structure of the skin, and thus, the micelles may
have been received by the skin barrier as hydrophobi c entities
that permit increased permeation (Willimann et al. 1992). It
should be pointed out that the micellar-borne drug showed higher
 TRANSDERMAL DELIVERY OF NICARDIPINE
permeability through the human stratum corneum than did the
corresponding gel to guinea pig skin. This could be explained
by the fact that organogel structure may be related closely to the
lipidic aggregates that exist in human skin and therefore acting
more as a skin penetration modiŽ er.
CONCLUSION
Vehicle effects on the percutaneous absorption of nicardip-
ine have been reported in the literature. Vehicles investigated in-
cluded blends of propylene glycol, trimethylene glycol, linoleic
acid, ethanol, water, and isopropyl myristate in the presence
or absence of a skin penetration enhancer. On the other hand,
the formulation components propylene glycol, oleic acid, and
dimethyl isosorbide strongly affect the permeation of nifedipine
resulting in an optimal  ux and lag time. The reported investiga-
tion discusses a novel method of administration of nicardipine
to promote transdermal absorption and provide a therapeutically
effective concentration of the drug in the blood-stream. We have
Downloaded by [Osaka University] at 22:10 15 July 2015
demonstrated that the choice of vehicle signiŽ cantly affects the
percutaneous absorption of nicardipine (as salt or base), through
the skin membrane. A binary mixture of PG and OA and a ternary
mixture of PG/OA/DMI possessed maximal  ux and minimal
lag times as compared with those obtained when pure solvents
were applied.
We concluded that using cosolvent systems whose compo-
nents function as carriers through the skin, therefore, could en-
hance transdermal absorption of nicardipine, and change the
value of the diffusion coefŽ cient and/or the activity coefŽ cient of
the drug in the skin barrier. We also revealed the skin penetration-
enhancing potential of a lecithin organogel, composed of IPM,
PG, OA, DMI, and water. In general, higher  uxes were observed
with nicardipine free base. In vitro human skin penetration stud-
ies of NC-base dissolved in the gel system resulted in a steady
state  ux of nicardipine (base) such that a transdermal device
with 7.3 cm in diameter should deliver the drug at rates that
provide clinically useful quantities.
REFERENCES
Bhatnagar, S., and Vyas, S. P. 1994. Organogel-base d system for transdermal
delivery of propranolol. J. Microencapsulatio n 11:431 – 438.
Curdy, C., Kalia, Y. N., and Guy, R. H. 2001. Noninvasive assessment of the
effects of iontophoresis on human skin in vivo. J. Pharm. Pharmacol. 53:769 –
777.
Diez, I., Colom, H., Moreno, J., Obach, R., Peraire, C., and Domenech, J. 1991.
A comparative in vitro study of transdermal absorption of a series of calcium
channel antagonists. J. Pharm. Sci. 80:931 – 934.
Dreher, F., Walde, P., Luisi, P. L., and Elsner, P. 1996. Human skin irritation stud-
ies of a lecithin microemulsion gel and of lecithin liposomes. Skin Pharmacol.
9:124 – 129.
247
Dreher, F., Walde, P., Walther, P., and Werhrli, E. 1997. Interaction of lecithin
microemulsion gel with human stratum corneum and its effect on transdermal
transport. J. Contr. Rel. 45:131 – 140.
Frishman, W. H. 1989. New therapeutic modalities in hypertension: Focus
on a new calcium antagonist-nicardipine . J. Clin. Pharmacol. 29:481 –
487.
Guy, R. H. 1998. Iontophoresis —Recent developments . J. Pharm. Pharmacol.
50:371 – 374.
Higuchi, T. 1960. J. Soc. Cosmet. Chem. 11:85. Cited by Seki, T., Sugibayashi,
K., and Morimoto, Y. 1987. Effect of solvents on the permeation of nicardipine
hydrochlorid e through the hairless rat skin. Chem. Pharm. Bull. 35:3054 –
3057.
Kou, J. H., Roy, S. D., Du, J., and Fujiki, J. 1993. Effect of receiver  uid
pH on in vitro skin  ux of weekly ionizable drugs. Pharm. Res. 10:986 –
990.
Mitragorti, S., and Kost, J. 2000. Low frequency sonophoresis : A noninva-
sive method of drug delivery and diagnostics. Biotechnol. Progr. 16:488 –
492.
Mitragorti, S., Edwards, D. A., Blankschtein, D., and Langer, R. 1995. A mech-
anistic study of ultrasonically-enhance d transdermal drug delivery. J. Pharm.
Sci. 84:697 – 706.
Potts, R. O., Mak, V. H. W., Guy, R. H., and Francoeur, M. L. 1991. Advance.
Lipid Res. 24:173 – 210.
Scartazzini, R., and Luisi, P. L. 1988. Organogel s from lecithin. J. Phys. Chem.
92:829 – 833.
Schurtenberger, P., Scartazzini, R., Majid, L. J., Leser, M. E., and Luisi, P. L.
1990. Structural and dynamic properties of polymer-like reverse micelles.
J. Phys. Chem. 94:3695 – 3701.
Seki, T., Sugibayashi, K., and Morimoto, Y. 1987. Effect of solvents on the
permeation of nicardipine hydrochlorid e through the hairless rat skin. Chem.
Pharm. Bull. 35:3054 – 3057.
Seki, T., Sugibayashi, K., Juni, K., and Morimoto, Y. 1989. Percutaneous ab-
sorption enhancer applied to membrane permeation-controlle d trandermal
delivery of nicardipine hydrochloride . Drug Des. Develop. 4:69 – 75.
Singh, S. K., Durrani, M. J., Reddy, I. K., and Khan, M. A. 1996. Effect of
permeation enhancers on the release of ketoprofen through transdermal drug
delivery systems. Pharma zie 51:741 – 744.
Sinha, V. R., and Kaur, M. P. 2000. Permeation enhancer s for transdermal drug
delivery. Drug Develop. Ind. Pharm. 26:1131 – 1140.
Sorkin, E. M., and Clissold, S. P. 1987. Nicardipine. A review of its pharma-
codynamic and pharmacokineti c properties and therapeutic efŽ cacy in the
treatment of angina pectoris, hypertension and related cardivascular disor-
ders. Drugs 33:296 – 345.
Squillante, E., Maniar, A., Needham, T. E., and Zia, H. 1998. Optimization of
in vitro nifedipine enhancemen t through hairless mouse skin. Int. J. Pharm.
169:143 – 154.
Squillante, E., Nanda, A., Needham, T. E., and Zia, H. 2000. Transdermal deliv-
ery of calcium channel blockers, such as nifedipine. U.S. Patent 6,106,856.
Squillante, E., Needham, T. E., Maniar, A., Kislalioglu, S., and Zia, H. 1998.
Codiffusion of propylene glycol and dimethyl isosorbide in hairless mouse
skin. Eur. J. Pharm. Biopharm. 46:265 – 271.
Willimann, H., Walde, P., Luisi, P. L., Gazzaniga, A., and Stropollo, F. 1992.
Lecithin organogels as matrix for transdermal transport of drugs. J. Pharm.
Sci. 81:871 – 874.
Yu, D., Sanders, L. M., Davidson, III, G. W. R., Marvin, M. J., and Ling, T.
1988. Percutaneous absorption of nicardipine and ketorolac in rhesus monkey.
Pharm. Res. 5:457 – 461.