1. PRESENTED BY
DIVYA SENGAR
(M.Sc. Biotech)
Under the supervision of
Dr. Deepak C. Chilkoti
(FORTIS CLINICAL RESEARCH, FARIDABAD)
2. The study was performed to evaluate the
pharmacokinetic parameters of the test
product to the reference product of the
Rizatriptan.
3. PHARMACEUTICAL EQUIVALENTS -
Pharmaceutical equivalents are drug products
that contain identical amounts of the identical
active drug ingredient, i.e., the same salt or
ester of the same therapeutic moiety, in
identical dosage forms, but not necessarily
containing the same inactive ingredients.
SCOPE OF THE GUIDELINES - Bioavailability and
Bioequivalence studies are required by
regulations to ensure therapeutic equivalence
between a pharmaceutically equivalent test
product and a reference product. Several in vivo
and in vitro methods are used to measure
product quality.
4. Bioavailability:
Bioavailability is the rate and extent to
which the active ingredient or active moiety
is absorbed from a drug product and
becomes available at the site of action.
Pharmacokinetics
o Absorption
o Food effect
o Distribution
o Metabolism and elimination
o Urinary Excretion
5. Migraine is a chronic neurological disorder
characterized by recurrent moderate to severe
headaches often in association with a number of
autonomic nervous system symptoms.
Typically the headache is unilateral (affecting one
half of the head) and pulsating in nature, lasting from
2 to 72 hours. Associated symptoms may
include nausea ,vomiting , photophobia, phonophobia
and the pain is generally aggravated by physical
activity.
Up to one-third of people with migraine headaches
perceive an aura.
Due to mixture of environmental and genetic factors.
Occurs more in women than men.
exact mechanisms of migraine are not known.
6. Rizatriptan benzoate is described
chemically as: N,N-dimethyl-5-(1H-
1,2,4-triazol-1-ylmethyl)-1H-indole-
3-ethanamine monobenzoate.
Its empirical formula is
C15H19N5•C7H6O2
white to off-white, crystalline solid
molecular weight of the benzoate
salt is 391.47
molecular weight of the free base is
269.4
Rizatriptan benzoate is soluble in
water at about 42 mg per mL
(expressed as free base) at 25°C.
oral, injectable, nasal spray , and
oral dissolving tablets.
7. Chemical and reagents –
DILUTION SOLUTION – 500 ml, of methanol and 500
ml of water were transferred into in a 1000 ml
reagent bottle. It was mixed well and sonicated at
ultrasonic bath. Solution was used the till next day
from its date of preparation and stored at room
temperature.
5mM Ammonium Acetate Solution - 385.40 ± 15.42
mg of ammonium acetate was transferred into a 1000
ml volumetric flask. 100 ml of water was added to
dissolve completely and volume was made up to the
mark with It was mixed well and sonicated at
ultrasonic bath. Solution was used the till next day
from its date of preparation and stored at room
temperature.
Rinsing Solution - Methanol
8. MOBILE PHASE - 800 ml of methanol and 200
ml of 5mM Ammonium Acetate solution was
transferred into 1000 ml reagent bottle. 1 ml
of formic acid was added and mixed well and
sonicated at ultrasonic bath. Solution was
used the till next day from its date of
preparation and stored at room temperature.
9. Admission and ICD presentation(N=31)
Written informed consent(N=27)
Enrollment(N=24+
02)
Standby discharged,subjects dosed in
period I
12 for test, 12 for reference
Sub 16 from test and sub 19 from
reference dropped out
Same process involved in period II
Completion of study(N=22)
10. Hematology Urinalysis Serology tests
Haemoglobin Physical examination HIV 1 & 2
Platelet count Colour HBsAg
Total leukocyte count pH HCV
Differential leukocyte count Specific gravity VDRL
Biochemistry Protein Additional tests
Blood urea nitrogen Glucose Alcohol breath test*
Serum creatinine Appearance
Total bilirubin Microscopic examination
ALP RBC
AST WBC
ALT Epithelial cells
Glucose Crystals
Cholesterol Casts
11. Sub. No. Age
(years)
Height
(cm)
Weight
(Kg)
BMI
(Kg/m²)
Diet * Gender Race Smoker Alcoholic
1 23 167.4 54.2 19.40 2 Male Asian No No
2 30 166.5 51 18.50 2 Male Asian No No
3 21 164.4 51.5 19.20 2 Male Asian No No
4 37 164.6 60.2 22.40 2 Male Asian No No
5 25 162.2 60.1 22.90 2 Male Asian No No
6 25 158.9 49.4 19.80 2 Male Asian No No
7 26 180.0 50.8 18.90 2 Male Asian No No
8 30 173.2 63.8 24.30 2 Male Asian No No
9 27 165.2 50.8 18.70 2 Male Asian No No
10 19 165 65.8 24.20 2 Male Asian No No
12. Wash out period – 7 days
Dosing , admission and stay
Treatments –
o Reference (R)
o Test (T)
Assignment to treatment sequences
Assessment of compliance
Sampling and separation
Meals to be given to subjects
13. A total of 44 blood samples (1 ×
05 ml each)
Pre-dose sample 10 ml.
Sample placed in wet ice- bath.
22 blood collection time points.
sample separation.
Stored below - 20˚C ± 10˚C
Transferred to the bioanalytical
section at FCRL for assay.
14. Food Item
Dinner (12 Hr To 10 Hr Before
Dosing)
Kala Chana Curry, Paneer Curry,
Curd, Chapati, Rice, Aloo Beans,
Kheer
Breakfast (0.5 Hr Before Dosing) Chicken, Whole Milk, Hash Brown
Potatoes, Fried Eggs, Bread Slice,
Butter
Lunch (4 Hr After Dosing) Dal Tdka, Aloo Paneer, Curd,
Chapati, Tomato Onion Salad
Snacks (9 Hr After Dosing) Samosa, Skimmed Milk
Dinner (13 Hr After Dosing) Green Gram Dal, Chapati, Cabbage
Peas Vegetable, Curd
Breakfast (24 Hr After Dosing) Skimmed Milk, Cornflakes, Sugar,
Bread Pakora, Apple
15. Dinner AWM
vitals
High
fat high
calorie
breakfa
st
Dosing AWM Vitals
AWM
AWM Vitals
lunch
Vitals
snacks
Vitals
dinner
AWM Vitals
dischar
ge
0900-
2230
│
-24hr
to
10.50
2100-
2300
│
-12hr
to 10
hr
↓
0730
│
-1.5 hr
↓
0830
│
-0.5hr
↓
0900
│
0hr
↓
1000
│
01hr
↓
1100
│
02hr
↓
1200
│
03hr
↓
1300
│
04hr
↓
1700
│
08hr
↓
2100
│
12hr
↓
0100
│
16hr
↓
0900
│
24hr
↑
Admission CE,
AWM & vitals
↑
Pre-dose
blood
sample
↑
Test (T)
or
Refere
nce (R)
↑
Safety
sample
in last
period
17. Volounteer compensation - The subjects
were adequately compensated on account of
their participation in the study as approved
from the institutional review board.
18. Rizatriptan, at doses up to and above the
projected clinical doses, was well
tolerated in this study. Although there
were reported occasional mild or
moderate symptoms of headache,
lightheadedness, dizziness, or fatigue.
Based on geometric mean ratio of log
transformed data and 90% confidence
interval of test and reference product
for Cmax, AUC0-t and AUC0-∞, it is
concluded that the ratio and extent of
absorption of test and reference product
are similar.
