Journal of Controlled Release 268 (2017) 159–165

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Journal of Controlled Release

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

Dihydroergotamine mesylate-loaded dissolving microneedle patch made of T

polyvinylpyrrolidone for management of acute migraine therapy
Cetin Tasa,b, Jessica C. Joycea,c, Hiep X. Nguyend, Padmanabhan Eangoord, Jennifer S. Knaackd,
Ajay K. Bangad, Mark R. Prausnitza,c,⁎
a
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA
b
Gülhane Education and Research Hospital, Department of Pharmaceutical Sciences, 06010 Etlik, Ankara, Turkey
c
Wallace Coulter Department of Biomedical Engineering at Georgia Tech and Emory University, Georgia Institute of Technology, Atlanta, GA 30332, USA
d
Department of Pharmaceutical Sciences, College of Pharmacy, Mercer University, Atlanta, GA 30341, USA

A R T I C L E I N F O A B S T R A C T

Keywords: Migraine is a widespread neurological disease with negative effects on quality of life and productivity. Moderate
Migraine to severe acute migraine attacks can be treated with dihydroergotamine mesylate (DHE), an ergot derivative that
Dihydroergotamine mesylate is especially effective in non-responders to triptan derivatives. To overcome limitations of current DHE for-
Microneedle patch mulations in subcutaneous injection and nasal spray such as pain, adverse side effects and poor bioavailability, a
Polyvinylpyrrolidone
new approach is needed for DHE delivery enabling painless self-administration, quick onset of action, and high
Liquid chromatography tandem mass
spectrometry
bioavailability. In this study, we developed a dissolving microneedle patch (MNP) made of polyvinylpyrrolidone,
Bioavailability due to its high aqueous solubility and solubility enhancement properties, using a MNP design previously shown
Rapid onset to be painless and simple to administer. DHE-loaded MNPs were shown to have a content uniformity of
108 ± 9% with sufficient mechanical strength for insertion to pig skin ex vivo and dissolution within 2 min. In
vivo pharmacokinetic studies were carried out on hairless rats, and DHE plasma levels were determined by liquid
chromatography-tandem mass spectrometry (LC-MS/MS). The area under curve (AUC) value after DHE delivery
by MNP (1259 ± 917 ng/mL min) was not significantly different (p > 0.05) as compared to subcutaneous
injection, with a relative bioavailability of 97%. Also, appreciable plasma levels of DHE were seen within 5 min
for both delivery methods and tmax value of MNPs (38 ± 23 min) showed no significant difference (p > 0.05)
compared to subcutaneous injection (24 ± 13 min). These results suggest that DHE-loaded MNPs have promise
as an alternative DHE delivery method that can be painlessly self-administered with rapid onset and high
bioavailability.

1. Introduction
Approximately 50% of adults worldwide suffer from an active
headache disorder, and according to a World Health Organization's
survey, headaches rank 19th among the most disabling conditions [1].
This sometimes-incapacitating health issue is among the top five most
disabling conditions for women [2]. Migraine headaches are the 3rd
most prevalent illness in the world and affect about 11% of adults
worldwide [3]. Attacks are often accompanied by one or more disabling
symptoms, and migraineurs consistently report reduced quality of life
between attacks [4]. The direct and indirect management of migraine
therapy has an annual economic cost of approximately $13 billion just
in the USA [5].
There have been many encouraging developments in antimigraine
medications over the past few decades, but currently available medical
therapies are still far from optimal. The introduction of the “triptans” in
the 1990s drastically changed prescribing patterns. Members of this
antimigraine drug family are considered the first choice for moderate to
severe attacks in migraine therapy unless there are contraindications
[6]. However, nearly one-third of patients taking triptans for acute
migraine therapy discontinue this therapy because of lack of efficacy,
migraine recurrence, cost, and/or side effects [7]. Thus, this subgroup
has dificulties managing acute migraine attacks and often seeks alter-
native drugs, such as dihydroergotamine mesylate (DHE) [8].
DHE is an ergot derivative that has been extensively utilized and
studied in the treatment of episodic and chronic migraine. The me-
chanism of action is most likely vasoconstriction by stimulating α-
adrenergic and 5-HT receptors [9]. Two pharmaceutical dosage forms
of DHE, namely parenteral and nasal spray, were approved in United
States in 1945 and 1990, respectively. An orally inhalable form of DHE

⁎
Corresponding author at: School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA.
E-mail address: prausnitz@gatech.edu (M.R. Prausnitz).

http://dx.doi.org/10.1016/j.jconrel.2017.10.021
Received 8 July 2017; Received in revised form 4 October 2017; Accepted 13 October 2017
Available online 16 October 2017
0168-3659/ © 2017 Elsevier B.V. All rights reserved.
C. Tas et al.
was submitted to FDA for approval in 2013, but is not yet approved
[10]. Nasal administration of DHE exhibits inconsistent pharmacoki-
netic (PK) profiles, resulting in poor acceptance among patients and
prescribers, whereas the parenteral route has the disadvantage of pain,
needle phobia, risk of infections at the injection site, and requirement of
specialized personnel for administration, which leads to poor patience
compliance [11]. Thus, there is a need for a new formulation design of
DHE that enables easy self-administration, safe and effective delivery
mimicking the parenteral (especially SC) PK profile. Such a new for-
mulation could offer convenience as well as therapeutic advantages for
migraineurs [12].
In the last decade, advances in the field of transdermal delivery
using microneedle patches (MNPs) have shown that the barrier function
of stratum corneum can be overcome while retaining the advantages of
patch-based delivery. Microneedles are typically a few hundred microns
in width and up to 1 mm in length, and they are arranged as an array on
a patch that is applied to the skin. The microneedles painlessly puncture
the stratum corneum and deliver drugs and vaccines to viable epidermis
and dermis below [13,14]. MNPs can be self-administered and are
strongly preferred over injection [15–17]. Among the different types of
microneedles, water-soluble polymer ones that dissolve in the skin have
received great attention because they do not generated sharps waste
and cannot be reused after removal from a patient's skin [18,19]. Prior
studies have investigated the delivery of triptans using microneedle
patches for possible treatment of migraine, including sumatriptan
[20–23], zolmitriptan [24] and rizatriptan [25].
It is desirable to have rapid uptake of anti-migraine drugs for fast
onset of relief to the patient. Rapid uptake is facilitated by the dense
capillary bed in the superficial dermis where microneedle patches de-
liver drug and by formulation with suitable polymeric excipients that
dissolve quickly, such as polyvinylpyrolidone (PVP) [26], fibroin [27],
maltose [28] and chondroitin sulphate [29].
The aim of this study is to formulate dissolving MNPs for rapid re-
lease and capillary uptake of DHE in the skin. We therefore used highly
water-soluble PVP as the microneedle matrix material to enable rapid
delivery to treat acute migraine therapy and used DHE as the active
pharmaceutical ingredient to provide relief to patients who suffer from
unsufficient medication with triptan derivatives. We carried out in vitro
tests measure content uniformity, mechanical strength of microneedle
needles for complete insertion into skin, solubility and delivery effi-
ciency of DHE from the MNPs, and optical microscopy imaging of
MNPs. We also conducted in vivo studies of bioavailability of DHE
delivered by MNPs in hairless rats. In this way, we introduce this al-
ternative therapy approach to acute migraine therapy with DHE-loaded
MNPs.
2. Materials and methods
2.1. Materials
DHE (Tocris, Minneapolis, MN, USA), caroverine HCI (Sigma-
Aldrich, St. Louis, MO, USA), PVP (10 kDa, Sigma-Aldrich) poly-
vinylalcohol (6 kDa, 78% hydrolyzed, Acros Organics, New Jersey,
USA), Sucrose (Fluka Analytical, St. Louis, MO, USA), Gentian violet
(Good Neighbor Pharma, Brawley, CA, USA), Optical Microscope
(Olympus SZX16, Shinjuku, Tokyo, Japan). All reagents were of ana-
lytical grade and used as received.
2.2. Fabrication of DHE-loaded MNPs
MNPs were prepared using two separate solutions. A stock solution
of DHE was prepared in methanol (adjusted to pH 4 with trifluoroacetic
acid, TFA) at a concentration of 30 mg/mL. This stock solution was
mixed with deionized (DI) water containing 10% w/v PVP to obtain a
final solution of 7.15 mg/mL DHE and 7.5% w/v PVP (i.e., the drug
solution). The backing solution consisted of polyvinyl alcohol, sucrose,
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Journal of Controlled Release 268 (2017) 159–165
and water in a mass ratio of 8:6:15. The production procedure consisted
of the following: (a) 25 μL of drug solution was cast onto a poly-
dimethylsiloxane (PDMS) microneedle mold, after which vacuum
(27 mm Hg) was applied to help pull the drug solution into the mi-
croneedle mold cavities; (b) excess drug solution was removed from the
mold surface with a flat blade; (c) the drug solution was allowed to dry
into the tips of the microneedle mold cavities; (d) approximately 200 μL
of backing solution was cast onto the microneedle mold under vacuum,
thus forming a complete microneedle patch after drying based on
methods published previously [30]. Each patch was then stored in a
desiccator at room temperature for 1 day before demolding with plastic
backing material (polymethylmethacrylate, McMaster-Carr, Elmhurst,
IL, USA). Demolded patches were stored in the desiccator until for
further analysis.
2.3. Assay of DHE in MNP
Each MNP loaded theoretically with 50 μg DHE was incubated into
50 mL DI water until dissolution was complete. The solution was then
filtered through a membrane filter having a pore diameter of 0.45 μm
and analyzed for DHE content with a validated HPLC coupled with
fluorescence detector. Briefly, chromatographic separation was per-
formed using a reverse-phase Zorbax C8 column (150 mm × 4.6 mm
i.d., 5 μm particle size, Agilent, Santa Clara, CA, USA). The mobile
phase consisted of acetonitrile and water (40:60 v/v) containing 0.1%
TFA and 0.1% triethylamine degassed prior to use. The column tem-
perature was 35 °C and the flow rate was set at 1.2 mL/min. The ex-
citation and emission wavelengths were 280 nm and 350 nm, respec-
tively. The total run time of each analysis was 6 min, and the retention
time of DHE was 4.2 min. The calibration curve was linear in the
concentration range 25–5000 ng/mL, and the correlation coefficient
was 0.999.
2.4. Ex vivo release of DHE from MNPs
DHE-loaded MNPs were inserted into pig skin ex vivo with thumb
force for 30 s. At predetermined time intervals (1, 2.5, 5 and 10 min),
MNPs were removed from the skin and the insertion site was tape-
stripped three times with an adhesive tape (3 M Transpore™, St. Paul,
MN) to remove residual DHE on the skin surface. Used MNPs were
imaged by optical microscopy to verify drug release into skin after in-
sertion and then assayed for residual DHE content. The amount of DHE
delivered into the skin was calculated by subtracting the amount of
DHE remaining in the MNPs after insertion and remaining on the skin
surface from the amount originally encapsulated in the MNPs.
To determine the residual amounts of DHE in the MNPs after skin
insertion, MNPs were dissolved in DI water. The stripped tapes were
soaked in methanol for 1 day at room temperature to recover the DHE
on the tape. The amount of DHE extracted from the MNPs and the
stripped tape was determined using HPLC, as described above.
2.5. In vitro dissolution of DHE from MNPs
DHE-loaded MNPs were incubated in a beaker containing 20 mL DI
water at 32 °C. At predetermined time intervals, 500 μL of dissolution
medium was removed and replenished with the same amount fresh DI
water. The samples were filtered through a membrane filter having a
pore diameter of 0.45 μm, and DHE content was analyzed by HPLC, as
described above.
2.6. Determination of DHE solubility
An excess of DHE was added to 5 mL of 10% w/v PVP in DI water,
sonicated for 1 h, and agitated in a shaker with a temperature main-
tained at 37 °C for 24 h. The suspension was filtered through a mem-
brane filter having a pore diameter of 0.45 μm, diluted with methanol
C. Tas et al.
and analyzed by HPLC with fluorescence detector. The average of three
experiments was taken. Solubility of DHE in DI water and in 10% w/v
PVP in DI water was found to be 0.64 ± 0.14 and 1.13 ± 0.19 mg/
mL respectively. The DHE solubility significantly increased by the ad-
dition of PVP (p < 0.05).
2.7. Insertion of DHE-loaded MNPs into pig skin ex vivo
Succesful insertion of DHE-loaded MNPs was evaluated on pig skin
ex vivo. Pig skin was carefully shaved with a razor to remove hair. DHE-
loaded MNPs were applied to the skin by pressing them down with the
thumb, left on skin for 1 min, and then removed. After that, the pierced
skin was stained with gentian violet for 5 min. Excess dye was removed,
and the skin was evaluated for the appearance of blue dots on the
stratum corneum using an optical microscope.
2.8. In vivo bioavailability studies of DHE loaded MNPs
All animal studies were conducted with approval by the Georgia
Institute of Technology Institutional Animal Care and Use Committee
(IACUC). Fifteen hairless male Sprague–Dawley rats (Charles River
Laboratories, Wilmington, MA, USA) weighing 500–550 g were equally
divided into three groups (i.e., five rats per group). Parenteral for-
mulations of DHE at a concentration of 1 mg/mL (containing alcohol,
glycerin and DI water as excipients at a pH value 3.6 ± 0.4) were
preparared under aseptic conditions. Group 1 received intravenous (IV)
injection of DHE parenteral formulation as a positive control. Group 2
received SC injection of DHE parenteral formulation as a positive
control. Group 3 received DHE by MNP delivery, where MNPs were left
on the skin for 30 min. The administered DHE dose was 50 μg for each
rat. The rats were anesthetized with isoflurane during drug adminis-
tration and until the end of the experiment. Blood samples (250 μL)
were collected by the jugular vein catheter using gel separator tubes
containing lithium heparin (Vacuette, Greiner Bio-One, Monroe, NC,
USA) at 0, 2, 5, 15, 30, 45, 60, 75, 90, 120, 150, 180, 240, 300 and
360 min after IV and SC administration and at 0, 5, 15, 30, 45, 60, 75,
90, 120, 150, 180, 240, 300 and 360 min after MNPs were first applied
to skin. All samples were centrifuged (Triac centrifuge, BD Diagnostic
Systems, Hunt Valley, MD, USA) at 3500 rpm for 5 min to collect
plasma.
2.9. LC MS/MS analysis
For analysis by LC-MS/MS, an Agilent 1200 series HPLC (Agilent)
and 6410B triple quadrupole mass spectrometer (Agilent) were used for
the study. A Waters XBridge C18 column (3.5 μm, 2.1 × 100 mm,
Waters, Milford, MA, USA) was used for chromatographic separation
and was maintained at 25 °C. Mobile phase A was comprised of HPLC-
MS grade water with 0.1% TFA and mobile phase B contained HPLC-MS
grade acetonitrile with 0.1% TFA. The run time was 23 min with a
gradient from 10% organic to 50% organic in 18 min at a flow rate of
0.5 mL/min. Ionization was performed using an electrospray ionization
source with the nebulizer pressure set to 35 psi, a gas flow rate of 12 L/
min, a source temperature of 300 °C and the capillary voltage set to
4000 V. Data acquisition was performed using Mass Hunter Data
Acquisition B.04.01 software (Agilent) under positive polarity. The m/z
transition of 584.3 → 270.2 was used to quantify DHE, and with the m/
z transition of 584.3 → 253.2 was used to qualify the data. The frag-
mentor voltage was set to 165 V and the collision energy was set to 30 V
for both the transitions of DHE. For the caroverine, internal standard,
the m/z transition of 366 → 100.2 was used as the quantitative ion and
366 → 121.2 was used as the qualitative ion. The fragmentor voltage
was set to 108 V for both the transitions of caroverine and the collision
energies were 24 V and 30 V for the quantitative and qualitative ions of
caroverine, respectively. Quantitative analysis was performed using
Mass Hunter QQQ Quantitative Analysis Software. A calibration curve
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Journal of Controlled Release 268 (2017) 159–165
was plotted with relative response (peak areas) of DHE to the internal
standard on the y-axis and the concentration (ng/mL) on the x-axis. The
unknown concentration of the samples was calculated by interpolating
their respective relative responses to the x-axis. The calibration curve
was linear in the concentration range 0.5–200 ng/mL, with a correla-
tion coefficient of 0.998.
2.9.1. Sample preparation
A 10 mg/mL caroverine stock solution and further dilutions of
caroverine were made using 50:50 acetonitrile:water. A 1 mg/mL DHE
stock solution was made in methanol. DHE calibrants of concentrations
0.5, 1, 5, 10, 50, 100 and 200 ng/mL, and quality controls of con-
centrations 2 ng/mL and 150 ng/mL, were made in rat plasma (Sprague
Dawley Rat Plasma, Innovative Research, Novi, MI, USA). Calibrants
and quality controls were prepared by serial dilution of the 1 mg/mL
stock solution of DHE in blank plasma.
The calibrants, quality controls and samples from the study were all
treated similarly. Aliquots (50 μL) of calibrants, quality controls and
samples were spiked with 10 μL of 50 ng/mL caroverine internal stan-
dard solution (previously diluted from the caroverine stock using 50:50
water:acetonitrile).
Liquid-liquid extraction was used to separate calibrants, quality
controls and samples after the addition of internal standard. Then, 5 μL
of 1 M ammonium buffer (5.35 g NH4Cl dissolved in 100 mL aqueous
ammonia) and 150 μL diethyl ether were added to the extracts and the
mixture was vortexed for 30 s before centrifuging for 5 min at 11,300 x
g. Approximately 150 μL of the supernatant was then pipetted out and
dried in a chemical hood for 1 h. After drying, samples were recon-
stituted in 50 μL of reconstitution solution (10:90 acetonitrile:water,
0.1% TFA).
2.10. Data analysis and statistical evaluation
Results are presented as the mean of n = 5 determinations with its
associated standard deviation (S.D.). Noncompartmental pharmacoki-
netic analysis was carried out using the Pharmacologic Calculation
System (version 4.1, Springer–Verlag, Philadelphia, PA, USA) computer
program which calculates the AUC (area under the curve) of the plasma
concentration as a function of time. The maximum plasma concentra-
tion (Cmax) and the time to reach the maximum plasma concentration
(tmax) were obtained from the experimental data. Absolute (Fabs; versus
IV) and relative (Frel; versus SC) bioavailability were calculated ac-
cording to following equations:
(AUCi × DoseIV )
Fabs = × 100%
(AUCIV × Dosei ) (1)
(AUCi × DoseSC )
Frel = × 100%
(AUCSC × Dosei ) (2)
where the subscript i corresponds to SC or MNP. All results are ex-
pressed as means ± S.D. Statistical differences between values were
determined using SPSS 24.0 for Windows software (IBM, Istanbul,
Turkey), with Student's t-test. The difference was regarded statistically
significant when p < 0.05.
3. Results
3.1. Characterization of DHE-loaded MNPs
MNPs were formulated using PVP as microneedle matrix material
because of its suitable properties, such as biocompatibility, water so-
lubility and rapid release of encapsulated drug after insertion into the
skin. A two-step casting process under vacum was used to localize the
drug in the tips of the needles, where the first cast was used to create
the drug-loaded microneedles and the second cast, which contained no
drug, was used to create the drug-free base of the microneedle array.
C. Tas et al.
Fig. 1. Microneelde patch for delivery of DHE. (a) Magnified view of a section of a re-
presentative microneedle patch. (b) A representative image of pig skin ex vivo after ap-
plication of a patch containing a 10 × 10 array of microneedles and stained with gentian
violet to identify sites of microneedle puncture into skin. (For interpretation of the re-
ferences to colour in this figure legend, the reader is referred to the web version of this
article.)
MNPs were fabricated as a 10 × 10 microneedle array in a ~1 cm2 area
attached to a clear supporting plastic backing (Fig. 1a).
MNPs formulated to contain 50 μg DHE were dissolved in 50 mL DI
water and DHE content was found as 108 ± 9% (Supplementary Table
S1). This shows that our lab-scale fabrication process was reproducible
in loading the target drug amount into each MNP.
3.2. Kinetics of in vitro dissolution of DHE from MNPs
The dissolution of DHE from MNPs was approximately 2 min when
submerged in DI water (Fig. 2a) due in part to the high water solubility
of PVP. The MNPs were designed for rapid dissolution of DHE in order
to expedite onset of action for rapid relief of migraine when used in a
future clinical senario.
3.3. MNP insertion and drug delivery into skin ex vivo
MNPs must have sufficient mechanical strength to pierce the skin
during application. MNPs were applied to pig skin ex vivo by pressing
on the patch backing with the thumb. These MNPs were designed for
manual insertion without the need for a high-velocity applicator. The
skin insertion site was then exposed to a violet tissue-marking dye that
selectively stains sites of skin puncture, which enabled visualization of
microneedle insertion efficiency. After staining the skin, a complete
array of violet spots (10 × 10) indicated that all microneedles were
162
Journal of Controlled Release 268 (2017) 159–165
Fig. 2. Dissolution profiles of DHE from microneedle patches. (a) Dissolution in DI water
as a receptor medium. (b) Delivery study in pig skin ex vivo. Data show mean ± SD
(n = 4).
succesfully inserted into the skin (Fig. 1b).
To assess drug delivery efficiency, the amount of drug deposited in
skin was measured and found to increase over time while the MNPs
remained on the skin. The amount of DHE delivered in skin was 62%
after 1 min and then steadily increased to 79% after 10 min (Fig. 2b).
There was very little drug deposited on the skin surface (1.4 ± 0.9%,
Supplementary Table S2). Imaging of MNPs after removal from skin
showed that the MNPs dissolved with similar kinetics, where the mi-
croneedle tips were dissolved within 1 min (Supplementary Fig. S1).
Because approximately 60% of the DHE loaded in MNPs was deposited
in skin within 1 min and the microneedle tips dissolved within the same
timeframe, these data indicate that most of the drug was localized to-
wards the tips of the needles, which dissolved quickly, probably due to
complete insertion into skin. Later time points up to 10 min only re-
leased an additional ~20% of the loaded drug, which indicates that this
drug was located towards the base of the microneedle, which may not
have been fully inserted into the skin and therefore dissolved more
slowly.
3.4. Pharmacokinetics and bioavailability of DHE in hairless rats
3.4.1. IV and SC injection
Parenteral formulations of DHE were first injected at a dose of 50 μg
(100 μg/kg) via IV and SC routes in hairless rats to determine the mean
plasma concentration of DHE versus time profile (Fig. 3). The average
peak concentration, Cmax, was 141 ng/mL and 11 ng/mL and the
average area under the curve, AUC, was 1751 ng mL/min and 1304 ng
mL/min for IV and SC injection, respectively (Table 1). There was a
significant difference between Cmax values after IV versus SC injection
(p < 0.05), but the difference between AUC values after SC and IV
C. Tas et al. Journal of Controlled Release 268 (2017) 159–165

Fig. 4. Representative optical microscopic image of a section of a microneedle patch

loaded with DHE after application to hairless rat skin in vivo for 30 min.

an alternative drug delivery route to SC injection. Imaging by optical

microscopy of MNPs after application to the rat skin shows that the tips
of the microneedles dissolved, with only the base part of MNPs re-
maining (Fig. 4). Measurement of residual DHE in used MNPs indicates
that the dose delivery efficiency of MNPs in vivo was 66% (Supple-
mentary Table S3), which is in reasonable agreement with ex vivo
findings (Fig. 2b).
We did not observe local skin irritation such as erythema or edema,
or any other adverse effects during the experiment.

4. Discussion

Migraine can cause significant negative effects on social activities

Fig. 3. Plasma concentration of DHE (a) after IV administration and (b) after MNP and SC
administration to hairless rats in vivo. Data show mean ± SD (n = 5).
injection was not significant (p > 0.05). The average time until peak
concentration, tmax, was 2 min after IV injection, which was sig-
nificantly faster than after SC injection, which was 24 (p < 0.01,
Table 1). Because the first data point after IV injection was taken at
2 min, it is possible that the tmax was even shorter and the Cmax was
even higher for IV injection than reported.
3.4.2. Delivery using MNPs
The pharmacokinetics of DHE delivery using MNPs showed that tmax
was not significantly different from SC injection (p > 0.05), whereas
that Cmax was significantly lower (p < 0.01, Fig. 3, Table 1). Cmax was
lower and tmax was longer for MNP delivery compared to IV injection
(p < 0.01, Fig. 3, Table 1). There was no significant difference of AUC
value after MNP delivery compared to SC injection (p > 0.05,
Table 1). Both MNP and SC delivery yielded AUC values significantly
lower than IV injection (p < 0.05, Table 1). The absolute bioavail-
ability of DHE after MNP delivery was 72%, and the relative bioavail-
ability after MNP delivery was 97%, which supports the use of MNPs as
and relationships, including decreased school and work attainment and
productivity [5]. There is a need for effective and acceptable migraine
treatments to decrease impact and disability associated with the dis-
ease. The formulation of DHE in MNPs can have a significant effect on
migraine therapy as a patient-friendly delivery method that serves as an
alternative approach for migraneurs, especially triptan non-responders
[31,32].
Migraine is one of the most common chronic illness, associated with
recurrent headache attacks and related symptoms in the gastro-
intestinal and autonomic nervous system [33]. Migraines affect ap-
proximately 18% of women and 6% of men in the United States and
nearly half of these people complain of reduced work or school pro-
ductivity [34]. Triptan derivatives are the first choice as the most ef-
fective therapy for acute migraine, but almost one-third of migraneurs
fail to achieve adequate pain relief and seek alternative medication to
alleviate migraine pain [12]. DHE is a semisynthetic ergot alkaloid
derivative with 5-HT1 agonist activity that is used to treat migraine.
Parenteral and nasal dosage forms of DHE are on the market, but dis-
advantages of these routes of administration have motivated research to
develop alternative dosage forms [35]. An ideal dosage form for mi-
graine treatment should have rapid onset of action, enable simple self-
administration, offer high bioavailability, and provide strong efficacy
and safety. While parenteral delivery has rapid onset with high bioa-
vailability, it is not simple to administer. Nasal delivery, in contrast, is
relatively simple to administer and offers rapid onset, but

Table 1
Pharmacokinetic parameters of DHE after application via different delivery routes in hairless rats (n = 5).

Delivery routea Formulation AUC (ng/mL·min) Cmax (ng/mL) tmax (min) Fabs %b Frel %c

IV Solution 1751 ± 873 141 ± 113 2

SC Solution 1304 ± 585 10.7 ± 2.3 23.8 ± 12.5 74,5 ± 33,4
Skin MNPs 1259 ± 917 7.1 ± 5.5 37.5 ± 22.6 71.9 ± 52.3 96.5 ± 70.3

a
IV: Intravenous; SC: subcutaneous.
b
Fabs: absolute bioavailability (relative to IV injection).
c
Frel: relative bioavailability (relative to SC injection).

163
C. Tas et al.
bioavailability is ~40% [36]. DHE administered as an oral inhaled
powder is a new pharmaceutical dosage form that has been under in-
vestigation for several years and offers fast pharmacologic response,
patient friendly use, and bioavailability of ~80% [12,37]. However, it
has not been approved by FDA due to technical concerns about the
manufacturing and functioning of the device [38].
In the last decade, there has been significant progress in the field of
transdermal drug delivery, notably in the development of MNPs that
increase skin permeability by creating micro-scale pathways across the
skin barrier and combine the simple self-administration of transdermal
patches and the delivery effectiveness of hypodermic needles [14,39].
MNPs have been shown to be suitable for many vaccines and drugs,
with clinical trials completed on delivery of zolmitriptan [40], para-
thyroid hormone [41], glucagon [42] and influenza vaccine [43,44].
MNPs have the advantages of painless self-administartion, no sharp
biohazardous waste, control over the drug delivery rate using for-
mulations, and cost-effective production technology [19].
In this study, we aimed to formulate DHE in MNPs as an improved
alternative to marketed parenteral and nasal dosage forms. PVP was
chosen as the microneedle matrix material to enable rapid and efficient
transdermal delivery of DHE [45]. PVP was chosen for four main rea-
sons. First, vinyl pyrrolidone monomer has a ring structure chemical
backbone that gives mechanical strength by increasing intramolecular
rigidity [26], which is important for insertion of microneedles into skin.
Second, PVP is higly water soluble at concentrations up to 50% [46]
which can enable rapid dissolution of encapsulated drug molecules
after insertion into skin. Third, PVP has been used as a plasma expander
clinically for decades [47]. Finally, PVP has been shown to increase
solubility of drugs that exhibit poor water solubility, such as acet-
aminophen and gidazepam [48]. The molecular weight of PVP used to
make MNPs was 10 kDa, because PVP with molecular weight < 20
kDa is known to be cleared effectively by the kidney, even after par-
enteral administration [47]. One drawback of PVP is that it is hygro-
scopic, and water absorption in a humid environment can reduce mi-
croneedle mechanical strength [49]. This issue was addressed by
storing MNPs with desiccant.
This study showed that DHE can be formulated in MNPs and ad-
ministered to hairless rats to generate a pharmacokinetic profile similar
to SC injection. The AUC (absolute bioavailability ~ 75%) and tmax
(~ 30 min) were not significantly different from each other with com-
parable Cmax (~ 10 ng/mL) which suggests that MNP administration
could be used to provide rapid relief from migraines with the kinetics of
injection, but without the pain and expertise needed to administer DHE
parenterally. Moreover the absolute bioavailability of DHE adminis-
tered by MNPs was much higher than nasal liquid formulations of DHE
reported previously, which was in the range of 16–50% in the rabbit
model [36].
Dose uniformity is also important for the development of MNPs
[50]. In this study, DHE dose uniformity in MNPs was 108 ± 9%. This
shows that even in our manual, laboratory-scale processes, each step in
fabrication such as preparing the drug solution with PVP, casting the
solution onto molds, applying vacuum and drying MNPs, was designed
and controlled to reach an acceptable dose uniformity range.
In this study, DHE delivery with MNPs had similar pharmacoki-
netics to SC injection, achieving a high relative bioavailability of 96%
(Table 1). This result is similar to bioavailability values determined for
MNPs loaded with exenatide [51] and insulin [18].
MNPs were fabricated with 50 μg DHE and administered the drug to
skin with a delivery efficiency of ~80% within 10 min ex vivo (Fig. 2b)
and ~ 65% within 30 min in vivo (Supplementary Table S3) with only
~ 1% of drug left on the skin surface (Supplementary Table S2). This
shows that MNPs can release the majority of loaded drug in a short
period of time, which reduces patch wearing time, thereby minimizing
risk of skin irritation related to long-term contact with transdermal
patches [52].
In prior studies, drug delivery efficiency from MNPs into skin ex vivo
164
Journal of Controlled Release 268 (2017) 159–165
has not always correlated with drug delivery in vivo. For example, in-
sulin formulated with starch gelatin in MNPs could be released into skin
within 5 min ex vivo, but in vivo studies in rats yielded an insulin tmax
value of 2 h [18]. This suggests that after application of MNPs to the
skin, insulin was entrapped in a gelatinous polymer matrix that slowed
its release and diffusion to capillaries for systemic distribution. In
contrast, MNPs formulated with sodium hyaluronate were able to de-
liver encapsulated exenatide with a tmax value of 15 min, which was
comparable to SC injection [51]. Like sodium hyaluronate, the PVP
used in our MNP formulation exhibits high water solubility, which may
explain the statistical equivalence of tmax after MNP and SC delivery.
However, further studies are needed to better determine tmax values,
because of large error bars and small number of animals used in this
study.
Drug molecules must dissolve and diffuse away from the delivery
site to reach the systemic blood circulation. Most drugs have either
weak acid or basic characteristics and exhibit poor aqueous solubility
[53]. Similarly, DHE is poorly soluble in water and can be classifed in
Biopharmaceutics Classification System as group II, which character-
istically exhibits low solubility and high permeability which typically
results in longer tmax values [54]. Increased drug dissolution at the
delivery site of absorption can lead higher bioavailability with shorter
tmax values. A common method to enhance drug solubility is to for-
mulate them in solid dispersion using the solvent evaporation method.
Solid dispersions are solid products consisting of at least two con-
stituents, typically a hydrophilic carrier and a hydrophobic active
substance. PVP, polyethylene glycols (PEG) and Plasdone-S630 are
commonly incorporated in solid dispersions as hydrophilic carriers
[55]. The production of DHE-loaded MNPs made of PVP involves a
process similar to the preparation of solid dispersions by solvent eva-
poration. The solubility enhancement mechanism of poorly soluble
drugs in solid dispersion may be related to increased area of solid-sol-
vent contact due to reduced particle size, improved wettability and
particles with higher porosity [56]. Thus, the similar pharmacokinetic
properties of MNPs formulated with PVP compared to SC injection may
be attributed in part to the MNP production process that resembles a
solid dispersion form of DHE.
5. Conclusion
DHE delivery using MNPs had a pharmacokinetic profile similar to
SC injection, as determined by the lack of statistically significant dif-
ference in tmax and AUC and with comparable Cmax values. DHE de-
livery was facilitated by formulation with PVP, which is believed to
enable rapid dissolution of microneedles due to the high water solu-
bility of PVP and to increase DHE solubility in a manner similar to solid
dispersions used in other types of dosage forms. These findings suggest
that DHE-loaded MNPs offer a promising new approach to treat acute
migraine attack with rapid onset of action, simple self-administration
and high bioavailability using a drug with well-established safety and
efficacy, which can represent a significant improvement over conven-
tions DHE delivery by parenteral and nasal routes of administration.
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.jconrel.2017.10.021.
Acknowledgements
Mark Prausnitz is an inventor of patents licensed to companies de-
veloping microneedle-based products, is a paid advisor to companies
developing microneedle-based products, and is a founder/shareholder
of companies developing microneedle-based products (Micron
Biomedical). This potential conflict of interest has been disclosed and is
managed by Georgia Tech and Emory University. Jessica Joyce was
funded through the NIH/NIGMS-sponsored Cell and Tissue Engineering
(CTEng) Biotechnology Training Program (T32GM008433) and
National Science Foundation Graduate Research Fellowship (DGE-
C. Tas et al.
1148903). The authors would like to thank Donna Bondy of Georgia
Tech for administrative assisstance. This work was supported by a grant
from The Scientific and Technological Research Council of Turkey
(TUBITAK) with the reference number of B.14.2.TBT.0.06.01-219-
19961.
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