Varaprasad Bobbarala et al.

/ Journal of Pharmacy Research 2009, 2(11),1751-1753

Research Article
ISSN: 0974-6943 Available online through
www.jpronline.info
A Randomised Bioequivalence Study on Vigrex in Twelve
Healthy Human Subjects under Fasting Conditions
Somasekhar Penumajji 1, Varaprasad Bobbarala2*, Rajesh Kumar Chadaram2, K. Chandrasekhar Naidu 3
1
Vivimed labs Limited, 2nd, 4th Floor, Veeranag towers, Habsiguda, Hyderabad, A.P.
2
For U Biosciences, A/4A, Park lane Residency, East point colony, Visakhapatnam-17, A. P,
3
Department of Botany, Andhra University, Visakhapatnam-530003, A.P. India.

Received on: 20-05-2009; Accepted on:15-07-2009

ABSTRACT

VIGREX (Sildenafil Citrate 50mg tablets) is a drug used to treat erectile dysfunction and pulmonary arterial hypertension (PAH). The single
dose randomized bioequivalence study was done to compare the pharmacokinetic parameters of VIGREX versus VIAGRA (Sildenafil Citrate
50mg tablets). After the test formulation of VIGREX is found to be bioequivalent with the formulation Reference of VIAGRA.

Keywords: VIGREX, VIAGRA, Sildenafil citrate, Cmax and Tmax.

INTRODUCTION
Vigrex (Sildenafil) is chemically designated as 1-[4-ethoxy-3-
(6, 7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo [4, 3-d] pyrimidin-
5-yl) phenylsulfonyl]-4-methylpiperazine. VIGREX is a drug used to
treat erectile dysfunction and pulmonary arterial hypertension (PAH)
Figure 1: Structure of Sildenafil
It acts by inhibiting cGMP specific phosphodiesterase type
5, an enzyme that regulates blood flow in the penis. Since becoming
available in 1998, sildenafil has been the prime treatment for erectile
dysfunction. The mechanism of action of Sildenafil citrate involves
the release of nitric oxide (NO) in the corpus cavernosum of the penis.
NO binds to the receptors of the enzyme guanylate cyclase which
results in increased levels of cyclic guanosine monophosphate (cGMP),
leading to smooth muscle relaxation (vasodilation) of the intimal cush-
ions of the helicine arteries, resulting in increased inflow of blood and
*Corresponding author.
Dr. Varaprasad Bobbarala
Scientist In-Charge,
For U Biosciences/IMMA Labs,
A/4A, Park lane Residency, East point colony,
Visakhapatnam, A.P-530017, India.
Tel.: + 91-9949129539
E-mail: varaprasadphd@rediffmail.com
an erection [1]. Sildenafil is a potent and selective inhibitor of cGMP
specific phosphodiesterase type 5 (PDE5) which is responsible for
degradation of cGMP in the corpus cavernosum. The molecular struc-
ture of sildenafil is similar to that of cGMP and acts as a competitive
binding agent of PDE5 in the corpus cavernosum, resulting in more
cGMP and better erections [1]. Without sexual stimulation, and there-
fore lack of activation of the NO/cGMP system, sildenafil should not
cause an erection. Sildenafil is metabolised by liver enzymes and ex-
creted by both the liver and kidneys. If taken with a high-fat meal,
absorption is reduced; the time taken to reach the maximum plasma
concentration increases by around one hour, and the maximum con-
centration itself is decreased by nearly one-third [2].
The primary indication of sildenafil is treatment of erectile
dysfunction (inability to sustain a satisfactory erection to complete
intercourse). It use is now standard treatment for erectile dysfunction
in all settings, including diabetes [3]. People on antidepressants may
experience sexual dysfunction, either as a result of their illness or as a
result of their treatment. A 2003 study showed that sildenafil improved
sexual function in men in this situation [4] Following up to earlier
reports from 1999 [5] the same researchers found that sildenafil was
able to improve sexual function in female patients on antidepressants
as well [6] As well as erectile dysfunction, sildenafil citrate is also
effective in the rare disease pulmonary arterial hypertension (PAH). It
relaxes the arterial wall, leading to decreased pulmonary arterial
resistance and pressure. This in turn reduces the workload of the
right ventricle of the heart and improves symptoms of right-sided
heart failure. Because PDE-5 is primarily distributed within the arterial
wall smooth muscle of the lungs and penis, sildenafil acts selectively
in both these areas without inducing vasodilatation in other areas of
the body. Pfizer submitted an additional registration for sildenafil to
the FDA, and sildenafil was approved for this indication in June 2005.
The preparation is named Revatio, to avoid confusion with Viagra,

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 Varaprasad Bobbarala et al. / Journal of Pharmacy Research 2009, 2(11),1751-1753
and the 20 milligram tablets are white and round. Sildenafil joins
bosentan and prostacyclin-based therapies for this condition [7].
Sildenafil has been shown to be useful for the prevention and treatment
of High altitude pulmonary edema associated with altitude sickness
such as that suffered by mountain climbers [8][9] While this effect
has only recently been discovered, sildenafil is already becoming an
accepted treatment for this condition, particularly in situations where
the standard treatment of rapid descent has been delayed for some
reason [10]. The objective of the single dose randomized
bioequivalence study was to compare the pharmacokinetic parameters
of VIGREX versus VIAGRA.
MATERIALS AND METHODS
The Drug controller General of India approved the study to
do at Mamata Hospitals and Diagnostics Center for Vivimed Labs
Limited. Twelve volunteers participated in a randomized, two treat-
ment, two way, two period, single dose crossover study. To be in-
cluded in this study, the subjects should be healthy Male volunteers,
21 to 35 years of age and within 15 percent of ideal body weight for
their height and build. One week prior to enrolment, a complete physi-
cal exam and biochemical screening was performed and history of
alcoholism or other drug abuse during the past year. Volunteers were
asked to abstain from taking any drug for at least one-week prior and
during the study, and to fast for at least 10 hours prior to the study.
They were asked to read and sign an informed consent, prior to the
study.
The study was divided in to two periods where the two
commercial trades Vigrex (sildenafil citrate 50mg) which is the test
sample and a reference sample Viagra (sildenafil citrate 50mg) were
orally taken in a randomized, double-blind, crossover order on each
Sunday with a washout period of 28 days. At 8:00 AM, a blood sample
was taken and each subject received a single oral dose of either brand
with 240 ml of water. Volunteers were remained fasting 2 hours after
the administration of the tablet. This was followed by a light break-
fast, standardized lunch and dinner 2, 6 and 12 hours after drug ad-
ministration, respectively. All subjects abstained from smoking and
exercise during the day of the study. Blood samples (5ml) were ob-
tained form the cubital and forearm veins through an indwelling hep-
arinized plastic tube, put into coded plain tubes 0.00 (predose), 0.25,
0.50, 0.75, 1.00, 1.25, 1.50, 2.00 2.50, 3.00, 4.00, 6.00, 9.00 12.00 and 24.00
post dosing. After the 24 hours sampling the volunteers were re-
leased. The withdrawn blood (5 ml) was placed in labelled tubes each
according to the volunteers’ Identification Number, time, and drug
code. Thereafter, these tubes were sent to the lab, and the plasma was
separated by centrifugation (within 30 minutes) at a rate of 5000 rpm
for 10 minutes. Plasma separated was stored at -70°C until the assay.
The stability of these samples at –70° C is three years. All samples
were analyzed in duplicates.
The plasma samples were analysed by reverse phase HPLC
for the content of Sildenafil Citrate, the pharmacokinetic parameters
were computed and subjected to statistical analysis. Sildenafil citrate
is extracted by taking 1.0mL of Plasma (containing Sildenafil) or stan-
dard or blank in a vial. In blank add 100ul of water. Then vortex the
samples to ensure complete mixing of contents. To each vial add
about 3.0mL of 2% isoamyl alcohol in hexane and shake for 10minutes
Journal of Pharmacy Research Vol.2.Issue 11.November 2009
by using extractor. Centrifuge for 15 minutes at 3500rpm. Transfer the
organic layer into another vial containing 0.3mL of 0.01N HCl and
shake for 10 minutes by using vortex mixer then centrifuge for 10min-
utes at 3500 rpm. Inject 50µl of acidic layer into chromatograph. The
chromatographic pattern has the mobile phase 0.5M KH2PO4 buffer:
0.0114M Diethyl ammonium chloride buffer: Acetonitrile (15:60:25)
with C18 column with a UV detection at 230nm.
For data analysis the chromatograms were acquired using
computer based software supplied by VP series, Japan. The concen-
tration of the unknown is calculated from the regression equation
using plasma calibration standard with the reciprocate of the drug
concentration as weighing factor (1/concentration). Y=mx+c where,
x=peak area ratio with internal standard peak, y=concentration,
m=slope of the calibration curve and c=x-axis intercept value.
Bioavailability is defined as rate and extent to which the drug reaches
from the site of application G.I tract to the site of measurement in the
body (plasma). Cmax and Tmax determine the rate at which the drug is
absorbed and was directly computed from the mean plasma concen-
tration curve. AUC0-t: Area under the plasma concentration time curve
from time 0 to t was obtained by the trapexoidal rule. K el is the elimina-
tion rate constant determined by linear regression of the last three
sampling points, T1/2 is determined by the formula T 1/2 = 0.693/ Kel. The
pharmacokinetic parameters were compared using paired test and
analysis of variance.
The following analysis was conducted on least square means
(LSM) of test and reference products using Win Nonlin Enterprise
software version 3.2. Analysis of variance was performed (a=0.05) on
the untransformed pharmacokinetic parameters AUC0-t, AUC0-a, Cmax
and Tmax . Additionally, log-transformed data was used for analysis of
AUC0-t, AUC0-a, and Cmax . The analysis of variance model included
sequences; subjects nested within sequence, period and drug formu-
lation as factors. The significance of the sequence effect was tested
using the subjects nested within the sequence as the error term.
Two-one sided t-test for Bioequivalence, with 90%confidence
intervals for difference between drug formulations were calculated
for AUC0-t, AUC0-a, and Cmax using both untransformed and log trans-
formed data. Power of ANOVA was calculated to detect as large or
greater than 20% of the reference mean. Ratio analysis was reported
for untransformed and log transformed AUC0-t, AUC0-a, and Cmax . The
geometric mean value was reported for log-transformed data.
RESULTS AND DISCUSSION
The test formulation showed a Tmax of 0.9797±0.167 as com-
pared to 1.0±0.151 of reference formulation. Thus the test and refer-
ence formulations have nearly same mean value. The least square
mean ratios for C max , AUC0-t and AUC0-a, are 104.58, 98.97 and 98.83 for
untransformed data and 100.60, 100.23 and 100.01 for log-transformed
data respectively indicating a comparable bioequivalence of test for-
mulation to the reference formulation. Their 90% confidence intervals
are 90.435-119.27, 89.309-115.677 and 87.093-114.69 respectively for
untransformed data and 91.244-118.352, 90.411-114.878 and 88.796-
115.688 respectively for log-transformed data. The 90% confidence
intervals are within the bioequivalence acceptance range of 80-120%
for the untransformed and 80-125% for the log-transformed data (as
per DCGI draft guidelines).
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Figure 2: Bioequivalence of Sildenafil Citrate
The test formulation of VIGREX (Sildenafil citrate 50mg tab-
lets), is bioequivalent with the reference formulation Reference
VIAGRA (Sildenafil citrate 50mg tablets).
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Source of support: Nil, Conflict of interest: None Declared
Journal of Pharmacy Research Vol.2.Issue 11.November 2009
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