Drug Development and Industrial Pharmacy, 2011; 37(6): 727–736

© 2011 Informa Healthcare USA, Inc.

ISSN 0363-9045 print/ISSN 1520-5762 online
DOI: 10.3109/03639045.2010.538061

RESEARCH ARTICLE

Strategies to improve dissolution and oral absorption

of glimepiride tablets: solid dispersion versus
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micronization techniques
Xiao Ning1,2, Jin Sun1, Xiaopeng Han1, Yue Wu1, Zhongtian Yan1, Jihong Han3, and Zhonggui He1
1
Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, P. R. China,
2
National Institute for the Control of Pharmaceutical and Biological Products, Beijing, P. R. China, and 3Institute for
Science and Technology in Medicine and School of Pharmacy, Keele University, Staffordshire, UK

Abstract
The objective of this study is to compare two different dissolution-enhancing strategies, solid dispersion (SD) and
micronized techniques, for improving oral absorption of poorly soluble glimepiride, and to decide which strategy is
suitable for its solubilization. The formulation of glimepiride SD was prepared by a solvent-evaporation process with
povidone k-30 (PVPk30) at a weight ratio of 1:9 (drug:carrier). The other was prepared via a modified micronization
technique, where glimepiride was premilled together with lactose and Lutrol F68 until the milled material passes
For personal use only.

through a 500 mesh ASTM sieve (30 μm). The dissolution results indicated that the two techniques were both capable
of enhancing the dissolution rate and extent of glimepiride. The release profiles of the two prepared products were
similar to the marketed product (Amaryl®) in various types of dissolution media. Furthermore, the oral bioavailability
was evaluated for the three formulations in fasted beagle dogs. Statistical analysis indicated that there were no
significant differences in pharmacokinetic parameters among the two prepared formulations and a marketed
product, especially for AUC0–36, Cmax, and Tmax. The dissolution parameters (D10 and AUC0–20) in Tris buffer demonstrated
the good in vitro/in vivo relationship with Tmax values for the three formulations. In conclusion, our studies confirmed
that both SD and micronization techniques were capable of improving dissolution and oral absorption of glimepiride
tablets to a similar extent as the marketed product, and the three glimepiride tablets were bioequivalent in the case
of the rate and extent of absorption in dogs.
Keywords:  Solid dispersion, micronization, glimepiride, in vitro dissolution, in vivo absorption

Introduction oral delivery formulation for poorly soluble drugs is now

For highly lipophilic compounds, it has been generally one of the most frequent and greatest challenges to for-
hypothesized that the uptake across the gut membrane mulation scientists in the pharmaceutical industry.
is efficient, resulting in sink conditions in the gut lumen, The most attractive option for increasing drug disso-
where the drug dissolves. In such case, the rate and extent lution is to improve the solubility through formulation
of absorption from the gastrointestinal tract (GIT) are approaches. Solid dispersion (SD) method is one of the
usually controlled and limited by its dissolution process. most commonly employed pharmaceutical approaches
Thus, dissolution rate and extent has presented a great to achieve the effect. Several methods can be employed
challenge to the development of suitable formulations for to manufacture a SD, such as melting1–3, dissolution in
oral administration of poorly water-soluble drugs. With a solvent4, or spray drying5, depending on the charac-
the recent advent of high-throughput screening of poten- teristics of the drug and carrier. In the present study,
tial therapeutic agents, the number of poorly soluble drug SDs with povidone k-30 (PVPk30) were prepared using
candidates has increased rapidly and the development of solvent-evaporation method. PVP belongs to a class of

Address for Correspondence:  Jin Sun, Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103 of
Wenhua Road, Shenyang 110016, P. R. China. E-mail: [email protected]; [email protected]
(Received 03 August 2010; revised 31 October 2010; accepted 01 November 2010)

727
 728  X. Ning et al.
water-soluble polymers, which has been used as a carrier decide which approach is better to prepare glimepiride
to increase the dissolution rate of various poorly water- tablets.
soluble drugs, such as ofloxacin6, glibenclamide7, and
carbamazepine8. From a chemical point of view, PVP is
Materials and methods
a polymeric lactam with an internal amide bond, which
contains a highly polar amide group that confers hydro- Materials
philic and polar-attracting properties. It is interesting Glimepiride was purchased from Chongqing Kangkeer
that it has biological structural features similar to those Pharm. Co. (Chongqing, China). Amaryl® 1 mg tablets
of proteins, and has a great potential for applications in were purchased from Aventis Pharma S.P.A. (Beijing,
the medical domain. China). Povidone k-30 (plasdone k-30) was obtained
Micronization technique is also a very promising from ISP Japan (Tokyo). PVPP (Polyplasdone® XL, USP
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approach to enhance dissolution rate and bioavailabil- grade) was a gift from ISP Japan (Tokyo). FLOWLAC® 100
ity of poor water-soluble drugs. It is well-known that (lactose monohydrate) was a gift from Meggle GmbH
particle size can affect the solubility of poorly soluble (Wasserburg, Germany). Avicel® PH 101 (MCC, micro-
solutes. In particular, a reduction in particle size may crystalline cellulose) was purchased from Asahi Kasei
improve the dissolution rate as a result of increased sur- (Tokyo, Japan). Lutrol F68 was kindly provided by BASF
face area in contact with the aqueous medium. Attempts (Ludwigshafen, Germany). Pregelatinized starch, sodium
have been made to modify the dissolution characteris- carboxymethyl starch, and magnesium stearate were
tics of fenofibrate9, phenacetin10, and oxcarbazepine11 by bought from Anhui Shanhe Pharm. S.P.A. (Anhui, China).
reducing particle size. However, a problem associated HPLC-grade methanol was purchased from Fisher
with the milling of a pure active ingredient is the forma- Scientific (Pittsburgh, PA). Water was prepared using an
tion of agglomerates. An alternative approach, adding EASYPURE®αRF/UV ultrapure water system (Barnstead
suitable excipients when milling, has been adopted to international Co., Boston, MA). All other materials were
address this problem in the present study. Other pos- of analytical grade and used as received.
sible methods include the use of amorphous forms to
increase drug solubility, the reduction of particle size Methods
to expand surface area for dissolution, and the reduc- Developments of the SD tablets (Formulation 1)
For personal use only.

tion of interfacial tension with the aid of a water-soluble

carrier. Preparation of physical mixture  Glimepiride and PVPk30
Glimepiride, 1-[[p-[2-(3-ethyl-4-methyl-2-oxo-3­- (passed through 100-mesh sieve, respectively) were
pyrroline-1-carboxami-do) ethyl]phenyl]sulfonyl]- accurately weighed and mixed in a sealed polyethylene
3-(trans-4-methylcyclohexyl) urea, is a new oral bag by hand for 10 min to obtain a homogeneous physi-
sulfonylurea hypoglycemic agent for the treatment of cal mixture (PM). The resulting mixtures were stored in a
non-insulin dependent (type II) diabetes mellitus. It desiccator at room temperature until further evaluation.
causes hypoglycemia by stimulating release of insulin
from pancreatic β cells and by increasing the sensitivity Preparation of SD by solvent evaporation  Accurately
of peripheral tissue to insulin. It also promotes the move- weighed amounts of glimepiride alone, PVPk30 alone,
ment of sugar from the blood into the cells that need it. and a series of mixtures of polymer and drug having a
Glimepiride is practically insoluble in water (2.7 × 10−4 final carrier–drug weight ratio ranging from 1:3 to 1:12
mg/mL at 25°C) and belongs to “Class II” drugs in the were dissolved at 40°C in minimum amount of CH2Cl2.
Biopharmaceutical Classification System12. It is likely to The solvent was evaporated under vacuum at 40–50°C.
display low and irregular bioavailability following oral Desiccation was completed in a vacuum oven until con-
administration due to the low solubility13,14. stant weight was achieved and the resulting solids were
It is reported that the product Amaryl produced by the pulverized. The dried powder was then passed through
Beijing Sanofi-Aventis, in comparison with the traditional a 100-mesh sieve and stored in a desiccator until further
glimepiride tablets, showed significant improvement evaluation.
in both in vitro dissolution and in vivo bioavailability15.
The tablet weight of Amaryl was 87 mg, and each tablet Differential scanning calorimetry analysis  Differential
contained 1 mg glimepiride. But regretfully, the manu- scanning calorimetry (DSC 60, Shimadzu) was used to
facturer has not yet reported its formulation and prepara- characterize the thermal properties of the drug, polymer,
tion technique in public. PMs, and SD. Ultrahigh purity nitrogen was used as the
In this work, we intended to develop two formulation purge gas at a flow rate of 150 mL/min. Samples weighing
tablets via different dissolution-enhanced strategies, SD 10 ± 5 mg were crimped in hermetic aluminum pans with
versus micronization, for the delivery of poorly soluble lids and then analyzed using a heating rate of 10°C/min.
glimepiride via oral route. With the commercial available The thermograms were recorded from 10°C to 250°C.
product Amaryl being the reference, the physicochemi-
cal and pharmacokinetic characteristics of the above two X-ray powder diffraction analysis  X-ray powder diffrac-
formulations were carried out and compared in order to tion analysis (XRPD) was performed using a D/Max-2400

 Drug Development and Industrial Pharmacy

 Strategies to improve oral absorption of glimepiride tablets  729
diffractometer (Rigaku Instrument, Japan). The samples and assayed for the dissolved drug concentration by
were exposed to Cu–Kα radiation under 40 kV and 60 mA HPLC. Each release test was carried out in the parallel
over the 2θ range from 3 to 45° in increments of 4°·per six times.
minute. The HPLC system consisted of an LC-10A HPLC
pump, a SPD-10A VP UV–VIS detector set at 228 nm, and
Tableting condition of Formulation 1  The SD with lactose, an ANASTAR interface (Tianjin, China). UV signals were
sodium carboxymethyl starch, PVPP, and magnesium monitored and peaks integrated using ANASTAR HSM
stearate was directly compressed to tablets (90 mg) by a software. A calibration was previously performed and
tablet-hitting pressure displacement system equipped it was confirmed that excipients produced no absorp-
with a flat-faced punch (6 mm diameter) (TDP single- tion signal at this wavelength across a wide range of
punch tablet press, Shanghai Pharmaceutical Factory, concentrations. Also, there was no interference in UV
Drug Development and Industrial Pharmacy Downloaded from informahealthcare.com by University Of Wisconsin Madison on 09/28/12

China). absorption between glimepiride and these excipients.

Chromatographic separations were performed at room
Preparation of micronized power tablets (Formulation 2) temperature using a C18 column (Hypersil BDS Silica,
4.6 mm × 150 mm, 5 μm; Phenomenex, China) guarded
Preparation of micronized PM  Co-grinding of glimepir- with a refillable precolumn (C18, 2.0 mm × 20 mm; Alltech
ide, Lutrol F68, and lactose (1:1:53, w/w/w) in a ball mill Associates, Inc., IL, USA) and a flow rate of 1.0 mL/min.
until the milled material passes through 200, 300, and The mobile phase consisted of methanol:0.01 M mono-
500 mesh ASTM sieves, respectively. ammonium phosphate buffer solution (pH 3.5) (70:30,
v/v) and was filtered through a 0.45-μm membrane filter
Tableting condition of Formulation 2  The micronized and degassed by ultrasonication before use. These con-
mixture with MCC, sodium carboxymethyl starch, PVPP, ditions resulted in a typical elution time for glimepiride
and magnesium stearate was compressed to tablets of 7.8 min.
(90 mg), by a tablet-hitting pressure displacement system To evaluate the dissolution behavior of test prepara-
equipped with a flat-faced punch (6 mm diameter) (TDP tions and marketed tablets in various medium simulat-
single-punch tablet press, Shanghai Pharmaceutical ing different physiology pH, dissolution was performed
Factory, China). in HCl of pH 1.2, water, phosphate buffer of pH 6.8, 7.2,
For personal use only.

and 7.8. Similarity index was introduced by Moore and

Solubility study Flanner in 199616 to determine similarity of two dissolu-
Excess samples equivalent to 1 mg glimepiride was tion profiles and is defined as follows:
added to 10 mL phosphate buffer of pH 7.8. The resulting
−0.5
suspensions were sonicated for 30 min and then shaken  n
2 
at 37°C for 24 h in a thermostatically controlled air bath, f 2 = 50 lg  1 + (1/ n ) ∑ ( Rt − Tt )  
 t =1 
since these conditions were previously shown to be suf-
ficient for achieving equilibrium solubility without any  where n is the sample number, and Rt and Tt are the per-
extra solubilization factor. Samples were then passed centages of the reference and test drug release, respectively,
through a 0.45 μm Millipore membrane filter, and the at different time intervals t. The f2 value is between 0 and
filtrates were suitably diluted and analyzed using HPLC. 100. The value is 100 when the test and the reference pro-
The HPLC method is described in the dissolution testing files are identical and approaches zero as the dissimilarity
section. increases, but because f2 is a log function small differences
in profile lead to a large drop in f2. If f2 of two dissolution
In vitro dissolution test drug release profiles is between 50 and 100, then these two
The dissolution test was carried out at 37 ± 0.5°C in drug release profiles are similar. Value under 50 indicates
100 mL of dissolution medium using a dissolution appa- differences between the release profiles17.
ratus (ZRS-8G Test Dissolution Tester, China). The test
was performed according to dissolution test method 2 In vivo bioavailability studies
as described in the Chinese Pharmacopoeia 2005 with a Experimental design  Beagle dogs were purchased from
paddle rotation speed of 75 rpm. The dissolution media General Hospital of Shenyang Military Region (China) and
used contained water, phosphate buffer of pH 6.8, 7.2, were given a normal standard chow diet and free access
7.8, and Tris solution (pH 10.4). The prepared tablets to water. Animals were housed in laminar flow house
(Formulation 1 and Formulation 2) and marketed maintained at 22 ± 2°C, 50–60% relative humidity under
Amaryl® weighed to be equivalent to 1 mg of drug were a 12-h light:12-h dark cycles throughout the experiment.
added to the dissolution apparatus, respectively, and Additionally, the dogs were kept in these facilities for at
2 mL of test fluid was withdrawn after 5, 10, 20, 30, 45, least 1 week prior to these experiments, and the stud-
and 60 min, and fresh dissolution medium was simul- ies were performed in accordance with the “Guiding
taneously replenished in the apparatus to maintain a Principles in the Use of Animals in Toxicology” adopted
constant total volume. The withdrawn samples subse- by the Society of Toxicology of US in July 1989 and revised
quently were filtered through a 0.45-μm Millipore filter in March 1999.

© 2011 Informa Healthcare USA, Inc.

 730  X. Ning et al.
The experiment involves fasting, single dose with
the three different preparations with washout period Relative AUC0−36 test
in between. Six dogs were used for each treatment = × 100
bioavailability (F%) AUC0−36 reference
group. The  dogs (13.1 ± 1.2 kg) were fasted for 24 h prior
to ­experiments and the preparations (Formulation 1,
Formulation 2, and reference product) containing 3 mg In vitro–in vivo correlations analysis
of drug were administered in the morning and a standard An in vitro–in vivo correlations (IVIVC) for glimepiride
lunch was given 4 h after dosing. Clinical recommenda- was evaluated by plotting the mean Tmax of the three
tion of human (50 kg) daily doses is 1 mg glimepiride. The formulations versus the mean in vitro accumulative dis-
animal dose was obtained by extrapolation of human daily solution at 10 min (D10) and AUC of 0–20 min (AUC0–20) in
dose based on body weight. Blood was withdrawn via can- various media, respectively. Linear regression analysis
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nulated needle from front legs. Four milliliters of blood was applied to fit the data and R2 was calculated to evalu-
were collected in heparinized tubes immediately prior ate the robustness of IVIVC.
to dosing (time zero) and at 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 5.0,
6.0, 7.0, 9.0, 12.0, 24.0, and 36.0 h after dosing. The plasma Results and discussion
was obtained by the centrifugation of blood at 3000 rpm
for 20 min and then kept frozen at−20°C until analysis. The
Phase solubility study of glimepiride PMs and SDs
The solubility phase diagram of glimepiride as a function
concentrations of glimepiride in plasma were determined
of PVPk30 proportion in phosphate buffer solution of pH
by ultra performance liquid ­chromatography-tandem
7.8 at 37°C is shown in Figure 1. For all the PMs and SDs
mass spectrometry (UPLC-MS/MS).
tested at different weight ratios, an increase in glimepir-
ide solubility was found linear with respect to the weight
Determination of glimepiride in dog plasma  A selective, fraction of polymer in the binary system. The increase of
rapid, and sensitive UPLC-MS/MS method was devel- the solubility with increasing PVPk30 concentration indi-
oped for the quantification of glimepiride in dog plasma. cates the solvent and wetting properties of PVP for drug.
With glipizide as an internal standard, sample pretreat- For example, at the highest PVP/drug ratio (12:1 w/w),
ment utilized a simple precipitating protein. A metha- the solubility of glimepiride was ~2-fold compared with
For personal use only.

nol solution of internal standard (400 ng/mL) 50 μL pure drug powder.

and 50 μL methanol were added to 50 μL plasma. After PMs exhibited an improvement in solubility over pure
vortex-­mixing for 3 min, the samples were centrifuged glimepiride coarse powder, maybe due to increased
at 15,000 rpm for 10 min. and the supernatant was col- wettability of the drug and the inhibition of drug par-
lected and centrifuged under the same conditions. The ticle aggregation by the polymer carrier. However, all
separation was carried out on an ACQUITY UPLCTM SDs enhanced solubility greater than their correspond-
BEH C18 column (50 mm × 2.1 mm, 1.7 μm; Waters Co., ing PMs. The enhancement of glimepiride solubility by
Milford, MA) with water (containing 0.3% formic acid) the SD technique over PMs was attributed to the wet-
and acetonitrile as the mobile phase at a flow rate of tability effect of the carrier and dispersed state of the
0.2 mL/ min. The detection was performed by a Waters drug. Furthermore, size reduction and molecularly/
Tandem Quadrupole (TQ) Detector (Waters). The mass
spectrometer was operated with electrospray ionization 20
(ESI) interface in positive ionization mode and with mul-
tiple-reaction monitoring mode. The selected reaction
Solubility of glim epiride(mg/L)

monitoring (SRM) of glimepiride and the internal stan-

dard were m/z 491 → 352 (glimepiride) and m/z 446.3  →
312.2 (glipizide), respectively. The concentration of
glimepiride was determined by a standard linear calibra- 15
tion curve in the concentration range of 25–5000 ng/mL.

Pharmacokinetic data analysis  Noncompartmental phar-

macokinetic analysis was conducted to calculate the area
under the curve from 0 to 36 h (AUC0–36). The peak plasma
concentration (Cmax) and the time to reach peak plasma 10
0 2 4 6 8 10 12
concentration (Tmax) of the different dosage forms were
PVP/drug ratio
determined by a visual inspection of the experimental data.
The AUC was estimated according to the linear trapezoidal solid disoersion physical mixture
rule. The threshold for differences to be considered sig-
Figure 1.  Effects of the weight fraction of PVPk30 on solubility
nificant was set at P 0.05. The ≤ relative bioavailability (F) of from glimepiride–PVPk30 systems in phosphate buffer solution
Formulation 1 and Formulation 2 to the commercial tablets of pH 7.8 at 37°C: physical mixtures (◆) and solid dispersions (◊).
(reference) was calculated using the following equation: Each point represents the means ± SD of six experiments.

 Drug Development and Industrial Pharmacy

 Strategies to improve oral absorption of glimepiride tablets  731
nanosized dispersed state of drug molecules in the SD Powdered glimepiride showed a melting endothermic
also accounted for the enhanced solubility. In addition, peak at 215.3°C, and PVP showed an endotherm at 202.7°C.
these results are in accordance with the well-established While these features are easily identified in the PM, the
formation of soluble complexes between water-soluble glimepiride endothermic peak disappeared totally. This
polymeric carriers and poorly soluble drugs18. suggests that the glimepiride was amorphous in SD.

Thermal analysis by DSC X-ray powder diffraction

DSC curves obtained for glimepiride, PVPk30, the PMs, XRPD is the standard technique for studying the crys-
and the SDs prepared with PVP are shown in Figure 2. talline or amorphous nature of drugs in solid state. The
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DSC
Detector: DSC60
mW
a

10.00
215.32°C

202.70°C c
69.29°C

0.00

d
69.28°C
For personal use only.

100.00 200.00
Temp[°C]

Figure 2.  DSC curves of glimepiride (a), physical mixtures (1:9, w/w) (b), PVP (c), and glimepiride–PVP solid dispersions (1:9, w/w) (d).

1600

1400

1200 a
Intensity(Counts)

1000

800
b
600

400 c

200
d

0
10 20 30 40 50
2-Theta(°)

Figure 3.  X-ray diffraction patterns of glimepiride (a), physical mixtures (1:9, w/w) (b), PVP (c), and glimepiride–PVP solid dispersions
(1:9, w/w) (d).

© 2011 Informa Healthcare USA, Inc.

 732  X. Ning et al.
Solid dispersion Physical mixture

100 100

80 80
Dissolution (%)

Dissolution (%)
60 60

40 40
Drug Development and Industrial Pharmacy Downloaded from informahealthcare.com by University Of Wisconsin Madison on 09/28/12

20 20

0 0
0 20 40 60 0 20 40 60
T/min T/min
1:3 1:5 1:7 1:9 1:10 pure drug Amaryl 1:3 1:5 1:7 1:9 1:10 pure drug Amaryl

Figure 4.  Dissolution profiles of glimepiride from solid dispersions or physical mixtures with various weight fractions of PVPk30 as a
function of time (min) in 0.02% Tris solution: (a) 1:3 w/w (◆); (b) 1:5 w/w (◼); (c) 1:7 w/w (▲); (d) 1:9 w/w (∆); and (e) 1:10 w/w (•) are the
drug–carrier ratio. Each value represents the mean ± SD of six experiments. For comparison: dissolution profiles of marketed Amaryl® 1 mg
tablets (——
 ) and pure glimepiride (—— ).

XRPD patterns for glimepiride, PVP, glimepiride–PVP SD

(1:9, weight ratio), and the corresponding PMs are shown
90
Dissolution percentage at 5min (%)
in Figure 3. In the X-ray diffraction spectrum of glimepir-
ide powder (Figure 3), sharp and intense peaks at a dif-
For personal use only.

fraction angle of 2θ 13.36°, 16.62°, 18.10°, 19.12,° 22.00°, 70

25.21°, 26.32° and a series of smaller peaks at 2θ angles
of 6.36°, 14.60°, 17.16°, 22.97°, and 23.64° were present 50
and suggested that the drug was present as a crystalline
material. Some glimepiride crystalline peaks were still
detected in the binary mixtures. In contrast, there were 30
no sharp peaks attributable to the crystalline form of
glimepiride in the SDs, suggesting that glimepiride in this 10
SD system was in amorphous state. 0 20 40 60 80 100
Percent of PVPK30 in sample (w : w)
In vitro dissolution testing
Investigation of dissolution in Tris solution Physical mixture Solid dispersion
The in vitro dissolution profiles of the SD-containing
Figure 5.  Effects of weight fraction of PVPk30 on the dissolution
preparations and corresponding PMs are shown in percentage at 5 min of glimepiride released from physical mixtures
Figure 4. Tris solution (pH 10.4) was selected as a dis- (◊) and solid dispersions (◆).
solution medium since it has been reported to have
sufficient discriminating capability to evaluate the of poor water-soluble drugs regardless of whether the
relationship between in vitro dissolution and in vivo drug is present as a PM or SD20. However, all SDs showed
absorption of glimepiride products19. The Formulation 1 faster dissolution rate than their corresponding PMs.
prepared by drug–PVPk30 SD at weight ratio of 1:9 and The enhanced dissolution rate of SDs may be attributed
1:10 (drug:carrier) exhibited rapid drug dissolutions, to many factors such as decreased particle size of drug,
with >75% drug released within 5 min, whereas pure specific dispersed state of drug in these SDs. In addition,
glimepiride coarse powder only released 30% of the drug PVP shows excellent water solubility. This polymer could
after 1 h. improve the wettability of the glimepiride particles and
The dissolution rates of glimepiride at 5 min from the drug solubility at the diffusion layer surrounding the
PM and SD with increasing proportion of PVPk30 are particles18. Furthermore, PVP would inhibit the crystal-
investigated in Figure 5. The established relationship lized process of the drug and then glimepiride was in the
revealed a nonlinear pattern of enhanced solubilization molecular dispersion in the SD as described earlier. It
and dissolution behavior. Indeed, there is a progressive was well-known that amorphous drug demonstrates the
increase in the dissolution rates of glimepiride up to 90% faster dissolution than crystalline drug21.
weight ratio of PVPk30 for PM and SD. These data indi- The dissolution rates of glimepiride attain the maxi-
cated that PVP is effective in improving the dissolution mal at weight fractions of about 0.9 of the polymer,

 Drug Development and Industrial Pharmacy

 Strategies to improve oral absorption of glimepiride tablets  733
evidencing that these ratios are crucial to achieve opti- the corresponding PMs of the same compositions. In con-
mal ­solubilization of glimepiride. For SDs, 81.4 ± 1.6% trast, the dissolution rate of glimepiride declines above
and 80.4 ± 2.3% of glimepiride dissolved within the first the drug to carrier ratio of 1:9. This feature may reflect
5 min in comparison with 35.8 ± 1.3% and 36.9 ± 2.4% for the delayed dissolution process of the drug induced by
the PVP in excess probably due to the relatively elevated
viscosity in the microenvironment around the drug
100 particle22. Moreover, the overall viscosity of the material
increased as the amount of PVP increases. The hardness
80 of the tablets was high so that disintegration delayed, and
ultimately affected the drug dissolution rate and extent.
Dissolution (%)

60 The results of this study demonstrated that 1:9 might

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be the most appropriate ratio to prepare the SD in

40 glimepiride–PVP binary system and was employed in the
optimal Formulation 1.
20 The in vitro dissolution profiles of the micronized
powder tablets (Formulation 2) were shown in Figure 6.
0 The enhancement of the dissolution rate from crude to
0 20 40 60 micronized glimepiride was in accordance with its pro-
T/min nounced reduction in particle size. The accumulative
Coarse powder 75µm 50µm 30µm Amaryl dissolution at 1 h was ~5-fold with particle size 10 μm
of glimepiride compared with coarse powder. This was
Figure 6.  Effect  of particle sizes of glimepiride on dissolution due to increased specific surface area of the drug as a
in 0.02% Tris solution: 10 μm (•), 50 μm (▲), 75 μm (◼), 145 μm
(◆) are the particle sizes of glimepiride. Each value represents consequence of reduced particle size. When particle
the mean  ± SD of six experiments. For comparison: dissolution size reduced to 30 μm, both in vitro dissolution rate and
profiles of marketed 1 mg tablets (——
 ). amount of glimepiride from tablets achieved ideal effect

Reference
For personal use only.

100

80
Dissolution (%)

60

40

20

0
0 20 40 60
T/min
PBS 6.8 PBS 7.2 PBS 7.8 Tris Water

Formulation 1 Formulation 2

100 100

80 80
Dissolution (%)

60 60

40 40

20 20

0 0
0 20 40 60 0 20 40 60
T/min T/min

Figure 7.  The dissolution profiles of glimepiride from optimal Formulation 1, Formulation 2, and reference products in phosphate
buffer solution at pH 6.8 (◼), 7.2 (▲), 7.8(▢), water (◆), and Tris (•) solution, respectively. Each data point represents the mean ± SD of six
measurements.

© 2011 Informa Healthcare USA, Inc.

 734  X. Ning et al.
in Tris solution and this condition was employed as the The similarity factors (f2) that were calculated between
optimal Formulation 2. the reference and Formulation 1/Formulation 2 are
It was worth mentioning that among the carriers used in presented in Table 1. The results of f2 showed that the
the formation of SDs, PVP is the most commonly used. This profiles of Formulation 1 are not significantly different
polymer shows excellent water solubility. When preparing from the reference except in phosphate buffer pH 7.2
the SD-based tablets, the difference between povidone as where the dissolution rate and extent of Formulation
the SD carrier and crospovidone as the ­disintegrant for 1 were both superior to the reference. Formulation 2
tableting purposes is the swelling ­capability. For example, and the commercial tablets are mostly similar except in
PVPP is a highly cross-linked version of PVP, exhibits water where Formulation 2 demonstrated substantial
excellent water-absorbing action, and swells very rapidly rapider ­dissolution. The results confirmed that both SD
to about 4–6 times their dry volume and sets up localized and micronization techniques are capable of improv-
Drug Development and Industrial Pharmacy Downloaded from informahealthcare.com by University Of Wisconsin Madison on 09/28/12

stresses that spread throughout the tablet and break up the ing dissolution of glimepiride to a similar extent as the
tablet from within. In contrast, PVP shows excellent water marketed product23,24. Based on the in vitro dissolution
solubility, improves the wettability of the poorly soluble performances, it was expected that the test and reference
drugs, but has poor swelling capability. products may be bioequivalent, but still needed further
bioequivalence verification.
Testing of solubility and dissolution in various media
To compare the dissolution behavior of optimal Pharmacokinetic studies
Formulation 1, Formulation 2, and marketed tablets, six Pharmacokinetics of the two optimal prepared tablets of
different dissolution media were investigated covering glimepiride (SD and micronized powder tablets) com-
the gastrointestinal physiological pH range of 1.2–7.8. The pared with the commercial available tablets were inves-
dissolution profiles for the three products are shown in tigated following oral administration of 3 mg to each of
Figure 7. The dissolution of the drug was strongly affected six healthy beagle dogs. The profiles of the mean plasma
by the pH, significantly greater dissolution at pH 7.8 than concentrations of glimepiride versus time are shown in
at pH 6.8. The dissolution at pH 1.2 was less than the limit Figure 9 and the main pharmacokinetic parameters are
of quantitation and cannot be detected. This observa- given in Table 2, respectively. Following oral administra-
tion was supported by the equilibrium solubility data tion of the two test formulations, absorption were both
For personal use only.

where a moderate increase was observed above pH 1.2 rapid with no statistically significant difference in mean
followed by a marked increase above pH 6.8 (Figure 8). Tmax when compared with the reference. The results
Glimepiride is a weak monoacid. The solubility of drug to suggested that SD and micronization techniques could
the pKa was calculated using the following equation: both improve drug release and absorption in GIT. This
observation was supported by the data as shown in Table
S − S0
pH = pK a +lg 3. Additionally, the results revealed that Formulation  1
S0 gave the highest Cmax value (2091.93 ng/mL, range
 where S and S0 are the equilibrium solubility and intrinsic
12
solubility of drug, respectively, and pKa is the dissociation
Solubility of glimepiride (µg/mL)

constant of drug. The pKa value of glimepiride has been

10
reported is ~6.5. Experimental results showed that there
was a poor solubility value of glimepiride under the pH 8
<6.5, and increased solubility with pH >6.5. For example,
the solubility was 0.1 μg/mL in pH 5.0 and 10.79 μg/mL 6
in pH 7.8.
4
Table 1.  Similarity factors (f2) of the three formulations in
different media. 2
Media Formulation 1 Formulation 2 Marketed tablet
Water 55.54 13.31 100 0
PBS6.8 55.16 59.70 100 0 2 4 6 8
PBS7.2 22.10 54.82 100 pH
PBS7.8 63.90 65.27 100
Figure 8.  Equilibrium solubility of glimepiride in different media
Tris 88.47 90.64 100 of pH values at 37°C.

Table 2.  Pharmacokinetic parameters (±SD) of glimepiride after oral administration of the three different formulations in beagle dogs,
including Formulation 1, Formulation 2, and marketed tablet.
Parameters Ke (h−1) T1/2 (h) Tmax (h) Cmax (ng/mL) AUC(0–t) (ng h/mL) F (%)
Marketed tablet 0.08 ± 0.03 9.45 ± 2.66 4.50 ± 1.02 1924.27 ± 776.76 26668.44 ± 7607.54 100.00 ± 0.00
Formulation 1 0.08 ± 0.02 8.64 ± 2.22 3.50 ± 1.97 2091.93 ± 875.47 28461.98 ± 9993.21 106.17 ± 16.66
Formulation 2 0.09 ± 0.02 8.63 ± 2.43 2.17 ± 0.68 1927.65 ± 752.75 26838.64 ± 8719.67 101.33 ± 16.26

 Drug Development and Industrial Pharmacy

 Strategies to improve oral absorption of glimepiride tablets  735
1254.80–3643.50 ng/mL), followed by Formulation  2 IVIVC analysis
(1927.65 ng/­mL, range 1345.73–3085.40 ng/mL) Since the level A of IVIVC in which the entire in vivo
and the ­reference (1924.27 ng/­mL, range 1269.91– time course is correlated with the in vitro data is diffi-
3364.05 ng/ mL), yet the differences are not regarded cult to acquire for the immediate-release dosage forms,
as substantial. The mean AUC value for Formulation 1 the level C of IVIVC that establishes a single-point rela-
was 28461.98 ng·h/­mL (range 17454.98–43666.89 ng·h/ tionship between dissolution and a pharmacokinetic
mL) and Formulation 2 was 26838.64  ng·h/mL (range parameter was developed in this study. Two dissolution
19492.24–41911.11 ng·h/­mL), whereas Amaryl® pro- rate parameters were selected, including D10 (cumulative
duced a mean of 26668.44  ng·h/mL (range 17138.68– release at 10 min) and AUC0–20 (area under the curve of
36065.39  ng·h/­mL). The ­relative bioavailability of the ­cumulative release from 0 to 20 min). Tmax was chosen
Formulation 1 and Formulation 2 against the reference to describe the in vivo absorption rate.
Drug Development and Industrial Pharmacy Downloaded from informahealthcare.com by University Of Wisconsin Madison on 09/28/12

calculated from AUC0–36 was 106.17% and 101.33%. Again, The regression analysis results are summarized in
no substantial differences between the test samples and Table  4. The R2 values between D10 in Tris buffer and pH  6.8,
reference were observed. Similarly, no differences in 7.2, and 7.8, with in vivo Tmax data, were 0.79, 0.02, 0.21,
half-life or elimination constant were found. It was noted and 0.26, respectively. Furthermore, R2 values between
that the inter-subject variability is relatively high, and so AUC0–20 in Tris buffer and pH 6.8, 7.2, and 7.8, with in vivo
another test including more subjects is necessary to eval- Tmax data, were 0.88, 0.02, 0.21, and 0.18, respectively. Both
uate the present results or clinic study to judge whether it data sets demonstrated that the dissolution performance
can be extrapolated to humans. of the three glimepiride formulations in Tris buffer cor-
related best with the in vivo Tmax data and can reflect the
in vivo dissolution behavior of the glimepiride tablets. On
3000.
the basis of IVIVC analysis, it was concluded that Tris solu-
Plasma concentration (ng/mL)

Reference tion was the good in vitro dissolution medium to screen

2500.
Formulation 1 and discriminate the different glimepiride formulations,
2000. and also the Tris solution was the optimum dissolution
Formulation 2
screening medium in the in vitro dissolution testing analy-
sis, such as Figures 5–7. Additionally, the results were in a
For personal use only.

1500.
good agreement with the results of the previous report19.
1000.

500. Conclusion
Compared with traditional glimepiride tablets prepared
0.
0 10 20 30 40
with pure drug coarse powder, the two formulation tab-
lets by SD and micronized powder showed significant
T/h
improvement in dissolution. Furthermore, the dissolution
Figure 9.  Average plasma concentration versus time profiles of profiles of the two new formulations are comparable with
glimepiride after oral administration (3 mg glimepiride doses) of the commercial product Amaryl. The pharmacokinetic
marketed tablet (◊); Formulation 1 (◆) and Formulation 2 (∆) in values for Cmax, Tmax, and AUC were similar for all formu-
beagle dogs (n = 6). Each data point represents the mean ± SD. lations with no statistical differences among them. These
data suggest that the investigated three formulations are
Table 3.  Wilconxon signed test results of Tmax between test A/B comparable based on in vitro characterization and are
and reference tablets for glimepiride. bioequivalent in terms of pharmacokinetic variables.
Reference Test A Test B P Conclusion Therefore, both the SDs and the micronization techniques
Mean ± SD 4.50 ± 3.02 2.17 ± 0.68 3.50 ± 1.97 >0.05 Not are able to improve the in vitro dissolution and in vivo
Max–min 9.00–1.00 3.00–1.50 6.00–2.00 substantial
bioavailability of glimepiride following oral administra-
Median 4.5 2 2.5 tion, to a similar extent as the marketed product Amaryl.

Table 4.  Regression analysis for in vitro and in vivo relationship.

Regression equation and correlation coefficient
Media D10a versus Tmax AUC0–20b versus Tmax
PBS 6.8 Y = –0.0125X + 3.7666 R2 = 0.0156 Y = –0.0009X + 3.8663 R2 = 0.0223
PBS 7.2 Y = –0.0301X + 5.206 R2 = 0.2051 Y = –0.0018X + 5.2537 R2 = 0.2122
PBS 7.8 Y = –0.1346X + 16.137 R2 = 0.2587 Y = –0.0083X + 16.757 R2 = 0.1808
Tris Y = –0.9536X + 94.503 R2 = 0.7893 Y = –0.0584X + 98.489 R2 = 0.8820
Water Y = –0.0322X + 4.3365 R2 = 0.7873 Y = –0.0018X + 4.3341 R2 = 0.7825
a
D10 is the cumulative release of the three studied formulations at 10 min.
b
AUC0–20 is the area under the curve of the cumulative release from 0 to 20 min.

© 2011 Informa Healthcare USA, Inc.

 736  X. Ning et al.

Declaration of interest processes. Part I: Micronization of neat particles. Int J Pharm 288:
3–10.
We are grateful for financial support from the 11. Douroumis D, Fahr A. (2006). Nano- and micro-particulate
National Basic Research Program of China (973 formulations of poorly water-soluble drugs by using a novel
optimized technique. Eur J Pharm Biopharm 63:173–175.
Program) No.  2009CB930300 and Key Project for Drug 12. Löbenberg R, Amidon GL. (2000). Modern bioavailability,
Innovation (No. 2010ZX09401-304, 2009ZX09301-012, bioequivalence and biopharmaceutics classification system. New
2008ZX09401-004) from the Ministry of Science and scientific approaches to international regulatory standards. Eur J
Technology of China. The authors report no declara- Pharm Biopharm 50:3–12.
tions of interest. 13. Grunenberg A, Keil B, Henck JO. (1995). Polymorphism in
binary mixtures as exemplified by nimodipine. Int J Pharm 118:
11–21.
14. Amidon GL, Lennernäs H, Shah VP, Crison JR. (1995). A theoretical
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