International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701

_____________________________________________________________________Research Paper

Development and Evaluation of Dienogest Vaginal Tablet

for Treatment of Endometriosis
Kailash Bansal*, Pankaj Pant, Rama Therdana Rao P, Kumud Padhee, Ajit
Sathapathy, Prithipal Singh Kochhar
Formulation & Development department, Jagsonpal Pharmaceuticals,
Rudrapur, Uttarakhand, India.

___________________________________________________________________________
Abstract

Vaginal delivery is an important route of drug administration for gynecological diseases. The
advantages of vaginal route are large surface area, rich blood supply, avoidance of first pass effect, relatively
high permeability of many drugs and self insertion. The aim of the present work was to develop Dienogest -
containing vaginal tablet for the treatment of endometriosis. Five different formulations were prepared, using
different compositions of Corn Starch, Microcrystalline Cellulose and Crospovidone. The tablets were evaluated
for hardness, friability, disintegration time, drug content and release. Formulation VT6 was found to be the best
among all formulations based on physical properties and in-vitro dissolution profile.

Keywords: Dienogest, endometriosis, in vitro release, vaginal tablet.

INTRODUCTION
As an important part of health of female
reproduction system, the vagina, in addition to
being a genital organ with functions related to
conception, serves as a potential route for drug
administration. [1,2] Endometriosis is a debilitating
gynecological medical condition in females in
which endometrial-like cells appear and flourish in
areas outside the uterine cavity, most commonly on
the ovaries. The uterine cavity is lined by
endometrial cells, which are under the influence of
female hormones. These endometrial-like cells in
areas outside the uterus are influenced by hormonal
changes and respond similarly as do those cells
found inside the uterus. Symptoms often worsen in
time with the menstrual cycle. Endometriosis is
typically seen during the reproductive years; it has
been estimated that it occurs in roughly 5% to 10%
of women. Symptoms may depend on the site of
active endometriosis. Its main but not universal
symptom is pelvic pain in various manifestations.[3]
It has been found desirable to use certain medicinal
agent like progesterone, progestational compounds
via vaginal route in the field of medical practice.
They have been administered in past via oral route
and found to yield certain undesirable side-effects.
These side-effects may be avoided by supplying
them directly to uterine wall.
Dienogest is an orally active synthetic steroid
with prominent progestational activity. It is thus
________________________________________
*Address for correspondence:
E-mail: kls.bansal@gmail.com
being evaluated clinically for treatment of
endometriosis, an estrogen-dependent disease.
Dienogest showed therapeutic effect in rats with
experimental endometriosis as well as amelioration
of endometrial implant-induced alteration in the
intraperitoneal immune system and maintenance of
bone mineral density, compared to commonly used
drugs for endometriosis. A long-term treatment
resulted in lower risk of thrombosis than
medroxyprogesterone acetate or danazol in
monkeys. Dienogest should thus serve as a highly
safe potent agent for treatment of endometriosis. [4]
Surgical and endocrine therapies successfully
suppress pelvic pain, but it often recurs after
completion of treatment. The pharmaceutical
preparation for treating endometriosis contains at
least 28, preferably 30, daily dose units, each of
which contain dienogest, cyproterone acetate, or
chlormadinone acetate at a daily dose that is at
most twice that required to inhibit ovulation
together with one or more pharmaceutical aids and
carriers. The daily dose units are administered in a
method of prophylaxis or therapy of endometriosis
continuously during a time interval of at least 169
days or 25 weeks, preferably more than two years.
The method effectively reduces endometriosis and
associated pain, while undesirable side effects
including bone density decrease are reduced or
eliminated. [5]
Although the suppositories are easily inserted,
they melt at body temperature and lead to
disturbing vaginal discharge. Oral gelatin capsules

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 International Journal of Research in Pharmaceutical and Biomedical Sciences
containing micronized progesterone have also been
used vaginally [6], but insertion of a small capsule
high into the vagina is difficult. Further, the rectal
route administration of progesterone is influenced
by first-pass effects of the liver and absorption is
highly variable, thus limiting the scope of rectal
route administration. Parentral administration
requires deep intramuscular injections which are
irritating at the site of injection and very painful.
The pharmaceutically effective amount of oral
dienogest required to inhibit ovulation is 1
mg/day.[7] The inhibition of ovulation by dienogest
occurs mainly via peripheral action as opposed to
central action on gonadotrophin secretion.[8] Oral
treatment of dienogest 2 mg/day in cyclical human
female reduces serum progesterone levels to
anovulatory levels; however serum levels of
lutenising hormone and follicle-stimulating
hormone are not significantly altered.[7]
Oral administration of dienogest results in adverse
effects including metrorrhagia (71.9%), headaches
(18.5%), and constipation (10.4%) [9]. So, a vaginal
route was proposed here for administration of
dienogest, a progestin of 19-nortestosterone
derivative, which is highly selective for
progesterone receptors, as an antifertility agent and
for effective treatment of endometriosis.
Micronized progesterone has been used in vaginal
administration for systemic use. [10] The vagina has
a large potential for absorption, and vaginal
administration results in higher uterine
progesterone concentrations. Continuous use of
vaginal progesterone does not influence the
hormonal, liver, or lipid profiles. There was no
reported case of endometrial hyperplasia. The
vaginal tablets of progesterone were found to be
well-tolerated, safe, and easily administered. [11]
The present invention is a solid dosage form of a
progesterone analog, i.e., dienogest which
disintegrates by a surface erosion mechanism. This
dosage form was developed for intravaginal use to
treat gynecological problems characterized by
progesterone deficiency, such as amenorrhea,
functional uterine bleeding, luteal phase defects,
premenstrual tension, and infertility, as well as in
osteoporosis therapy. Physical and pharmacokinetic
properties of drug Dienogest is given in Table No.
1 was done using talc and magnesium stearate, for 5
The literature review indicates that Dienogest
has not been focused for the local treatment of
vaginal disease as yet. It was, therefore, proposed
to prepare Dienogest tablets 2 mg to deliver
biologically effective amount of drug within one
hour, after insertion into the low moisture
environment of human vagina and lowers the side
effects of the oral use of Dienogest Tablets. The
present invention provides several advantages over
the prior art, including:
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ISSN: 2229-3701
1. Availability of an Active Pharmaceutical
Ingredient to vaginal epithelium and
mucosa directly.
2. Chances of less side effects and more
patient compliance.
MATERIAL AND METHOD
Dienogest was obtained as a gift sample from
Indo Phyto Chemicals Pvt. Ltd. (Nainital, India).
Lactose Monohydrate was purchased from DMV-
Fonterra (Netherland). Corn Starch was purchased
from Roquette, France. Microcrystalline Cellulose
was procured from FMC Biopolymer, Ireland.
Crospovidone and Povidone were purchased from
ISP Sales (UK) Ltd. Talc was purchased from
Luzenac, Italy. Magnesium Stearate was purchased
from Ferro Corporation, USA. All the chemicals
were of analytical grade and Purified Water was
used throughout the experiment.
EXPERIMENTAL
Particle Size Reduction:
Dienogest drug was micronized using a Air Jet
Mill (Promas India, 2” model). This exercise was
performed to check effect of micronization on in-
vitro dissolution profile of drug from the
formulation.
Preparation of Vaginal Tablet:
Different formulation trials are listed in Table
2. First Dienogest, lactose monohydrate, corn
starch and crospovidone were co-sifted through a
#40 mesh, geometrically. A solution of PVP K-
29/32 was prepared in Purified water. The
geometrically mixed blend was granulated with
PVP K-29/32 solution so as to achieve desired
consistency of wet mass, in a Rapid Mixer
Granulator (Make- Gansons, India). This wet mass
was then dried in a Fluidized Bed Dryer (Make-
Retsch TG 200, Germany) to get the desired
moisture content. Sifted the dried granules through
#24 mesh. The oversized granules were milled in a
Multimill (Make- Gansons, India) on medium
speed and with knife forward. Blended milled
granules with the remaining granules. Lubrication
minutes in an octagonal blender (Make- Ganson,
India). The granules were then compressed using a
rotary compression machine (Make- Lab Press,
India).
Characterization and Evaluation of prepared
formulations:
Particle Size Determination:
Particle size distribution of Dienogest before
and after micronization was checked by Malvern
Method (Table No. 3, Figure No. 3 and 4).
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 International Journal of Research in Pharmaceutical and Biomedical Sciences
Granule Properties:
The granule properties were evaluated with the
help of Tapped density tester (Electrolab, India)
using USP I method. The Bulk Density, Tapped
Density, Compressibility Index and Hausner’s
Ratio were calculated. The results are demonstrated
in Table No. 4.
Evaluation of Tablets:
The prepared tablets were evaluated for
Hardness (Electrolab, India), friability (Roche
friabilator), weight variation, in vitro disintegration
time and drug content. In vitro dissolution studies
of the vaginal tablets were performed by using USP
type II apparatus at 75 rpm and Simulated vaginal
fluid (pH 4.2), 1000 ml volume was used as
dissolution media. Temperature of dissolution
media was maintained at 37⁰C ± 0.5⁰C. Aliquot of
dissolution media were withdrawn at a specific
time interval and it was filtered. Absorption of
filtered solution was checked by UV spectroscopy
and drug content was determined from standard
calibration curve. The results for the evaluation of
tablets are shown in Table no.5.
Hardness:
The tablet hardness, the force required to break
a tablet in a diametric compression force, was
carried out using hardness tester from Electrolab,
India.
Friability:
The friability of the tablets was measured using
Roche friabilator (Electrolab, India). 6.5 gm tablets
were weighed and then rotated at 25 rpm for 4
minutes. Tablets were reweighed after dedusting
(removal of fines) and the percentage of weight
loss was calculated. Friability below 1.0% w/w was
considered acceptable.
In vitro disintegration test:
The disintegration time of the tablet was
measured in water (37±2⁰C) according to
disintegration test apparatus USP. The time (in
minutes) taken for complete disintegration of the
tablet with no palpable mass in the apparatus was
recorded. Six tablets from each batch were tested
for the disintegration time calculation.
Content Uniformity (By HPLC method):
Twenty tablets were weighed and powdered.
An amount of the powder equivalent to 2 mg of
Dienogest was transferred to 250 ml volumetric
flask, add 170 ml Diluent (HPLC grade water and
methanol in ratio 50:50 v/v repectively), sonicate
for 20 minutes and make up the volume with
diluent. Centrifuge the above solution at 2500 rpm
for about 5 min.
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Chromatographic Details (For Dienogest
Estimation):
Mobile phase: Mix water and Acetonitrile in the
ratio of 600:400 v/v respectively. Filter through
0.45µm membrane filter and degas it.
Diluent: Acetonitrile, Mixture of HPLC grade
water and methanol.
Chromatographic system:
Column : Thermo Hypersil
BDS, 150 x 4.6 mm, 5µm
Column temperature : Ambient
Flow rate : 1.0 mL/minute
Injection volume : 50 µl
Detector Wave length : 305 nm
Run Time : 8 minutes
In vitro dissolution profile:
The release profile of Dienogest vaginal tablets
was determined by using USP type I dissolution
test apparatus at 75 rpm and 37⁰C±0.2ºC
temperature. Five ml of aliquots were withdrawn at
predetermined intervals and filtered. The required
dilutions were made with Simulated Vaginal Fluid
pH 4.2 and the solutions were analyzed for the drug
content by HPLC spectrophotometer against
suitable blank. Equal volume of the dissolution
medium was replaced in the vessel after each
withdrawal to maintain sink condition. From this
the percentage of drug released was calculated and
plotted against function of time to study the pattern
of drug release.
ACCELERATED STABILITY STUDIES
The optimized formulation (VT6) was charged
on stability at 40ºC/75% RH for 1 month, 3 months
and six months. After completion of specified time
period in storage conditions, these stability samples
were subjected to drug content and dissolution
studies. The result of the study is mentioned in
Table No. 7.
RESULT AND DISCUSSION
The present work is directed to a tablet for
insertion into human vagina, which will then
disintegrate in the fluid present. The optimization
of disintegration times for vaginal tablets is quite
different from more traditional theoretical and
practical approaches that are utilized for oral
dosage forms. The different amount of moisture
present in mouth and vagina are primarily
responsible for the different approaches required in
these two regions. A high quantity of super
disintegrants can break the tablet in a few seconds
in moist environments like mouth and vagina. The
optimized batch was selected on the basis of low
Disintegration time. The tablets of Batch VT6 were
found to have lowest Disintegration time, i.e., 50
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 International Journal of Research in Pharmaceutical and Biomedical Sciences
seconds among all the formulations. The tablet
readily absorbs fluid to disintegrate rapidly in the
limited volume of fluid, pH 3.5-4.5, generally
present in the vagina, by virtue of its ‘wicking
effect’ that transports water to the tablet’s interior,
and by virtue of its high surface to volume ratio.
The tablet is preferably formulated to erode in the
vagina within one hour. In-vitro dissolution data
supports this claim (Table No. 6). The eroding
tablet produces micronized particles of the drug
which are dispersed in the vagina as a milky
suspension along with the other ingredients in the
tablet.
The Simulated Vaginal Fluid pH 4.2 [12] was
selected to study the release of Dienogest Vaginal
tablets because the prevailing pH of vagina is
maintained by lactobacilli which produce sufficient
lactic acid to acidify vaginal secretions to pH 3.5-
4.5.[13]
The Active pharmaceutical ingredient
(Dienogest) was micronized in an Air Jet Mill (Fig.
3 and Fig. 4) and the particle size of micronized
API was found to be 7.36 µ which finally helped in
enhancing Dissolution rate (Table No. 3). The
prepared granules showed good flow properties and
the results are shown in Table No. 4.
All the tablet formulations showed acceptable
phycochemical properties and complied with the
range specified by US Pharmacopeia 33 for
Disintegration, weight variation, friability and
hardness (Shown in Table No.5).
The concentration of Povidone was kept
constant in all the formulations while the
concentration of Lactose monohydrate, Corn
Starch, Microcrystalline cellulose PH 101 and
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ISSN: 2229-3701
crospovidone were varied batch to batch. As the
concentration of Crospovidone was increased, the
disintegration time decreases subsequently.
In formulation VT1, unmicronized Dienogest API
was employed which shows lower in-vitro
dissolution rate while all other formulations (VT2,
VT3, VT4, VT5 and VT6), developed with
micronized Dienogest API showed good
dissolution profile. Out of all the formulations VT6
showed best in-vitro release.
Assay of the optimized batch (VT6) was carried
out by the HPLC method and was found to be
99.85%. Stability studies revealed that there was no
significant change in appearance, assay, and drug
release profile at 40⁰C with 75% RH. (shown in
Table no.7).
CONCLUSION
Formulation VT6 shows the desired physical
parameters, disintegration time and dissolution
profile. The clinical effectiveness of the prepared
formulation is yet to be performed. However, it
would be necessary to conduct more studies like
long term clinical trials with large number of
patients, comparative studies with other modes of
treatment (oral administration of Dienogest Tablet
2 mg, vaginal administration of gel of Dienogest
Tablet 2 mg) and formulations to assess its
therapeutic efficacy. This non-mucoadhesive
Dienogest Tablet 2 mg is likely to be well received
by gynecological professionals in future because of
its non-stickiness and good in-vitro dissolution and
likely effectiveness in-vivo.
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TABLES
Table No. 1: Drug Properties

S. No. Parameters Remarks

1. Description White or slightly yellowish-white powder.

2. CAS No. 65928-58-7

3. Structural Formula

4. Molecular Weight 311.42 g/mol

5. Molecular Formula C20H25NO2

6. Chemical Name 17α-cyanomethyl-17β-hydroxy-estra-4,9-di-ene-3-one

7. Melting Point 209º-214ºC

8. Bioavailability 90%

9. Protein binding 90%

10. Metabolism Hepatic

11. Half life 6-12 hours

12. Excretion Renal

TABLE 2: Formulation Chart

Batch No. VT1 VT2 VT3 VT4 VT5 VT6

Ingredients Quantity per tablet in mg

Dienogest 2.0* 2.0** 2.0** 2.0** 2.0** 2.0**
Lactose Monohydrate 73.0 73.0 72.0 69.0 68.0 68.0
Corn Starch 10.0 10.0 10.0 35.0 35.0 35.0

Microcrystalline Cellulose 60.0 60.0 60.0 37.0 37.0 36.0

Crospovidone 1.0 1.0 2.0 2.0 3.0 4.0
Povidone 3.0 3.0 3.0 3.0 3.0 3.0
Purified Water*** q.s. q.s. q.s. q.s. q.s. q.s.

Talc 1.0 1.0 1.0 1.0 1.0 1.0

Magnesium Stearate 1.0 1.0 1.0 1.0 1.0 1.0
Total 150.0 150.0 150.0 150.0 150.0 150.0

*Unmicronized Dienogest was used.** Micronized Dienogest was used.***Does not appear in the final product.

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Table No. 3: Particle Size Distribution Data

Drug Batch No. Micronization Particle size Particle size Particle size
Substance Status Distribution Distribution Distribution
[D10%] [D 50%] [D 90%]
Dienogest AP/DNG/171 Unmicronized 1.95 µ 10.90 µ 45.97 µ

Dienogest AP/DNG/171 Micronized 1.09 µ 3.21 µ 7.36 µ

Table No. 4: Granule Properties

Granule Properties Formulations

VT1 VT2 VT3 VT4 VT5 VT6

Bulk density (gm/cc) 0.62 0.47 0.60 0.62 0.60 0.49

Tapped density (gm/cc) 0.81 0.62 0.79 0.79 0.77 0.61

Compressibility index 23.21 23.96 24.18 21.03 21.93 20.71

Hausner’s ratio 1.30 1.31 1.32 1.27 1.28 1.26

Table No. 5: Physical Properties of Dienogest Vaginal Tablets

Formulation Code Average Thickness Diameter (mm) Hardness (N) Friability (At 100 Disintegration Time
Weight (mg) (mm) revolutions)

VT1 150 2.93-2.99 7.03-7.05 42-48 0.15% 5:30 minute

VT2 150 2.90-2.94 7.03-7.05 40-50 0.17% 5:20 minute

VT3 150 2.90-2.96 7.02-7.05 40-45 0.22% 6 minute

VT4 150 2.95-3.01 7.00-7.03 39-43 0.18% 3 minute

VT5 150 2.94-2.98 7.01-7.06 43-47 0.20% 2 minute

VT6 150 2.94-2.99 7.02-7.05 42-48 0.25% 50 seconds

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 International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701

Table No 6: Dissolution parameters of compressed tablets.

Cumulative % Formulation Code

Drug release
VT1 VT2 VT3 VT4 VT5 VT6

10 min 68 82 84 87 89 93

15 min 74 84 85 88 90 95

20 min 78 86 87 90 92 96

30 min 82 88 90 92 94 97

45 min 85 90 93 93 95 98

60 min 87 92 94 96 97 99

Table 7: Accelerated Stability Study Data

40ºC/75% RH (Formulation No. VT6)

Test Initial 1 Month 3 Month 6 Month

Assay 99.85% 99.80% 99.60% 99.50%

Dissolution Profile

Time (minutes) 0 0 0 0

10 93 89 89 87

15 95 92 90 88

20 96 94 93 90

30 97 96 95 93

45 98 97 95 94

60 99 98 97 96

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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701

100

80

Cumulative % drug release

VT1
60 VT2
VT3
40 VT4
VT5
20
VT6
0
0 10 20 30 40 50 60
Time (min)

Fig 1: Dissolution profile of all the Formulations in Simulated Vaginal Fluid pH 4.2

100
Cumulative % drug release

80

60
Initial
40 One month
Three month
20
Six month
0
0 10 20 30 40 50 60
Time (min)

Fig 2: Dissolution profile of Formulation VT5 at initial condition compared with Stability at
40⁰C/75%RH for 1 month, 3 months and 6 months.

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Fig 3: Particle Size Determination (PSD) report of Dienogest (Unmicronised)

Fig 4: Particle Size Determination (PSD) report of Dienogest (Micronised)

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