Experiment 1 –

Graded Dose-Response Curve

B1
24 July 2007
Outline
• Introduction
• Objectives
• Materials and Methods
• Results
Expected
Actual
• Discussion
• Conclusion
DOSE-EFFECT RELATIONSHIP
• The intensity and duration of a
drug’s effects are a function of
the drug dose and drug
concentration at the effect site

• X-axis – drug concentration

• Y-axis – response
DOSE-RESPONSE CURVE
TYPES OF DOSE-RESPONSE RELATIONSHIPS
GRADED DOSE-RESPONSE QUANTAL DOSE-RESPONSE

• Continuous scale (↑dose → ↑ • All-or-none pharmacologic

effect) effect
• Measured in a single biologic • Population studies
unit
• Relates dose to frequency of
• Relates dose to intensity of effect
effect
DOSE-RESPONSE CURVE
POTENCY
• Relative position of the dose-
response curve along the dose
axis
MAXIMAL EFFICACY
• Determined principally by the
properties of the drug and its
receptor-effector system
• Reflected in the plateau of the
dose-response curve
• Shows the clinical
effectiveness of a drug
OBJECTIVES
• GENERAL
To compare the anti-inflammatory properties of aspirin,
paracetamol, celecoxib, and prednisone using the grade
dose-response curve.

• SPECIFIC
To determine the percent (%) response to inhibition of
inflammation with aspirin, paracetamol, celecoxib and
prednisone on Carrageenan-induced foot edema of rats.
To compare the relative potency and maximal efficacy of
aspirin, paracetamol, celecoxib, and prednisone anti-
inflammatory using the graded dose-response curve.
MATERIALS AND METHODS
• 12 week old rats
• Phlogistic Agent
(Carrageenan)
• Gavage Tubes, Syringe
• Mercury Set-up
• Drugs
▫ Paracetamol
▫ Aspirin
▫ Celecoxib
▫ Prednisone
Doses : per 200 g rat
Carrageenan
• phlogistic agent of choice for testing anti-
inflammatory agents both steroidal and
nonsteroidal
• is a mucopolysaccharide derived from Irish sea
moss, Chondrus
Katzung. Basic and Clinical Pharmacology, 10th ed.
COX-2 Inhibitors Pathway

CELECOX
IB
PARACETAMOL
• Weak COX-1 and COX-2 inhibitor due to
activity in the brain
• No significant anti-inflammatory effects; more
on analgesic effects
RECENT STUDIES:
• selective inhibition of the enzyme COX-3
in the brain and spinal cord
ASPIRIN
• Anti-platelet (160-325mg/day)
▫ Inhibits synthesis of TXA

• Antipyretic (325-650mg/4 hrs)

▫ Inhibit PGE2

• Anti inflammatory (4-6gday)

▫ COX-1 and COX-2
CELECOXIB
• Selective COX-2 inhibitor
▫ Effective anti-inflammatory drug
PREDNISONE
• Glucocorticoids
• anti-inflammatory and immunosuppressive
effect
• inhibits Phospholipase A2
• COX-2 inhibiting effects as the NSAIDs
% difference (inflammation)
= (reading after - reading before injection of phlogistic agent)
reading before injection of phlogistic agent
% inhibition
= (reading before injection of phlogistic agent - reading after )
reading before injection of phlogistic agent
RESULTS
Expected
▫ Anti-inflammatory property (Efficacy)
Prednisone > Celecoxib =Aspirin > Paracetamol
▫ Potency
Prednisone > Celecoxib > Paracetamol > Asprin
RESULTS
Actual
▫ Anti-inflammatory property (Efficacy)
Celecoxib =Aspirin > Prednisone> Paracetamol
▫ Potency
Prednisone > Celecoxib > Paracetamol = Asprin
SOURCES OF ERROR
• Variability in handling
• Initial inflammation
• Drug was not fully dissolved in NSS
• Spillage during gavage feeding
• Measurement of foot volume
Mercury level
Reader variability
 Graded-Dose Response Curve SECTION A Graded-Dose Response Curve SECTION B

Paracetamol Foot Volume Aspirin Foot Volume

Preparati Caragee Preparati Carageen
on: 500 mg tablet Before nan 1 hr 2 hrs 3 hrs on: 200 mg tablet Before an 1 hr 2 hrs 3 hrs
0.5 mg / 200 0.5 mg / 200
Doses: gm rat Doses: gm rat
Rat 1 0.002 0.002 0.005 0.004 0.004 Rat 1 0.002 0.002 0.002 0.001 0.004
Rat 2 0.002 0.002 0.006 0.005 0.005 Rat 2 0.002 0.002 0.005 0.001 0.003
Rat 3 0.003 0.001 0.007 0.006 0.007 Rat 3 0.001 0.003 0.001 0.001 0.001
Rat 4 0.002 0.003 0.004 0.004 0.005 Rat 4 0.003 0.003 0.002 0.001 0.001
Rat 5 0.002 0.004 0.005 0.002 0.006 Rat 5 0.001 0.001 0.003 0.001 0.002
Rat 6 0.002 0.003 0.005 0.004 0.005 Rat 6 0.001 0.001 0.008 0.001 0.003
Rat 7 0.001 0.004 0.004 0.005 0.007 Rat 7 0.002 0.005 0.002 0.001 0.001
Rat 8 0.001 0.008 0.005 0.004 0.003 Rat 8 0.001 0.001 0.001 0.001 0.001

1.6 mg / 200 1.6 mg / 200

gm rat gm rat
Rat 1 0.002 0.003 0.006 0.002 0.007 Rat 1 0.004 0.002 0.004 0.003 0.004
Rat 2 0.002 0.005 0.005 0.002 0.004 Rat 2 0.003 0.003 0.002 0.002 0.004
Rat 3 0.002 0.004 0.005 0.005 0.005 Rat 3 0.004 0.004 0.004 0.002 0.002
Rat 4 0.002 0.004 0.004 0.005 0.004 Rat 4 0.002 0.003 0.001 0.001 0.001
Rat 5 0.002 0.004 0.005 0.006 0.005 Rat 5 0.003 0.005 0.001 0.002 0.002
Rat 6 0.001 0.001 0.003 0.005 0.005 Rat 6 0.002 0.005 0.003 0.003 0.001
Rat 7 0.002 0.003 0.006 0.004 0.003 Rat 7 0.003 0.004 0.003 0.003 0.001
Rat 8 0.001 0.004 0.007 0.005 0.008 Rat 8 0.003 0.005 0.001 0.003 0.001

5 mg / 200 gm 5 mg / 200 gm
rat Rat 1 0.001 0.001 0.003 0.004 0.003 rat Rat 1 0.002 0.003 0.002 0.002 0.002
Rat 2 0.001 0.003 0.003 0.004 0.006 Rat 2 0.003 0.003 0.002 0.001 0.002
Rat 3 0.001 0.002 0.005 0.007 0.005 Rat 3 0.002 0.003 0.002 0.001 0.001
Rat 4 0.001 0.006 0.007 0.004 0.005 Rat 4 0.003 0.002 0.002 0.001 0.001
Rat 5 0.001 0.004 0.003 0.003 0.005 Rat 5 0.002 0.003 0.005 0.005 0.001
Rat 6 0.002 0.002 0.001 0.003 0.004 Rat 6 0.005 0.003 0.003 0.004 0.002
Rat 7 0.001 0.006 0.006 0.005 0.004 Rat 7 0.006 0.007 0.005 0.004 0.003
Rat 8 0.001 0.002 0.004 0.002 0.006 Rat 8 0.004 0.005 0.002 0.003 0.002

16 mg / 200 gm 16 mg / 200
rat Rat 1 0.001 0.001 0.004 0.008 0.005 gm rat Rat 1 0.003 0.001 0.001 0.001 0.001
Rat 2 0.002 0.004 0.005 0.004 0.005 Rat 2 0.001 0.002 0.007 0.004 0.001
Rat 3 0.001 0.002 0.003 0.005 0.007 Rat 3 0.004 0.001 0.001 0.001 0.001
Rat 4 0.002 0.002 0.005 0.004 0.005 Rat 4 0.002 0.001 0.004 0.001 0.001
Rat 5 0.001 0.004 0.004 0.005 0.005 Rat 5 0.002 0.002 0.001 0.002 0.001
Rat 6 0.001 0.003 0.005 0.006 0.003 Rat 6 0.001 0.001 0.001 0.001 0.008
Rat 7 0.001 0.002 0.007 0.006 0.004 Rat 7 0.001 0.001 0.001 0.002 0.001
Rat 8 0.001 0.003 0.005 0.004 0.005 Rat 8 0.001 0.001 0.001 0.004 0.006
 Graded-Dose Response Curve SECTION D
Graded-Dose Response Curve SECTION C
Prednisone Foot Volume
Celecoxib Foot Volume Preparat Caragee
Prepar Carageena ion: 20 mg tablet Before nan 1 hr 2 hrs 3 hrs
ation: 200 mg tablet Before n 1 hr 2 hrs 3 hrs 0.02 mg / 200
0.3 mg / 200 Doses: gm rat
Doses: gm rat Rat 1 0.004 0.001 0.006 0.002 0.005
Rat 1 0.001 0 0.007 0.002 0.003 Rat 2 0.004 0.005 0.005 0.003 0.003
Rat 2 0.002 - - - - Rat 3 0.002 0.004 0.005 0.004 0.005
Rat 3 0.001 0.005 0.006 0.003 0.006 Rat 4 0.004 0.005 0.005 0.003 0.002
Rat 4 0.004 0.003 0.004 0.003 0.002 Rat 5 0.001 0.004 0.006 0.002 0.005
Rat 5 0.001 0.006 0.002 0.006 0.006 Rat 6 0.001 0.001 0.006 0.003 0.002
Rat 6 0.002 0.004 0.003 0.003 0.001 Rat 7 0.003 0.003 0.005 0.001 0.003
Rat 7 0.002 0.003 0.002 0.003 0.004 Rat 8 0.002 0.006 0.002 0.002 0.001
Rat 8 0.002 0.007 0.006 0.003 0.002
0.06 mg / 200
1 mg / 200 gm rat
gm rat Rat 1 0.002 0.004 0.005 0.003 0.007
Rat 1 0.004 0.006 0.003 0.001 0.007 Rat 2 0.001 0.006 0.005 0.002 0.001
Rat 2 0.001 0.004 0.004 0.002 0.003 Rat 3 0.004 0.001 0.004 0.004 0.006
Rat 3 0.006 0.004 0.002 0.005 0.001 Rat 4 0.002 0.005 0.002 0.002 0.001
Rat 4 0.003 0.005 0.005 0.001 0.006 Rat 5 0.001 0.005 0.005 0.002 0.005
Rat 5 0.003 0.005 0.005 0.001 0.001 Rat 6 0.002 0.005 0.004 0.006 0.004
Rat 6 0.005 0.006 0.004 0.005 0.005 Rat 7 0.002 0.002 0.005 0.005 0.003
Rat 7 0.003 0.007 0.003 0.001 0.004 Rat 8 0.003 0.003 0.005 0.005 0.003
Rat 8 0.004 0.008 0.004 0.003 0.003

0.2 mg / 200 gm
3 mg / 200 rat Rat 1 0.003 0.003 0.001 0.005 0.005
gm rat Rat 1 0.003 0.007 0.004 0.003 0.003 Rat 2 0.001 0.004 0.002 0.005 0.002
Rat 2 0.005 0.006 0.006 0.004 0.002 Rat 3 0.002 0.004 0.006 0.004 0.002
Rat 3 0.004 0.005 0.005 0.003 0.002 Rat 4 0.002 0.003 0.005 0.004 0.004
Rat 4 0.005 0.007 0.003 0.002 0.001 Rat 5 0.004 0.006 0.001 0.004 0.004
Rat 5 0.001 0.006 0.003 0.004 0.002 Rat 6 0.002 0.003 0.005 0.004 0.005
Rat 6 0.002 0.005 0.004 0.003 0.004 Rat 7 0.001 0.005 0.005 0.004 0.001
Rat 7 0.006 0.007 0.008 0.004 0.003 Rat 8 0.001 0.003 0.006 0.004 0.003
Rat 8 0.005 0.004 0.003 0.004 0.001

0.6 mg / 200 gm
10 mg / 200 rat Rat 1 0.003 0.007 0.002 0.005 0.005
gm rat Rat 1 0.003 0.007 0.006 0.005 0.003 Rat 2 0.001 0.003 0.006 0.005 0.004
Rat 2 0.004 0.005 0.006 0.004 0.002 Rat 3 0.003 0.003 0.006 0.004 0.002
Rat 3 0.006 0.005 0.005 0.002 0.002 Rat 4 0.001 0.003 0.003 0.006 0.006
Rat 4 0.005 0.005 0.005 0.003 0.006 Rat 5 0.003 0.006 0.005 0.006 0.004
Rat 5 0.005 0.004 0.006 0.003 0.004 Rat 6 0.001 0.005 0.005 0.007 0.006
Rat 6 0.005 0.006 0.003 0.003 0.003
Rat 7 0.003 0.003 0.005 0.003 0.004
Rat 7 0.007 0.005 0.005 0.003 0.002 Rat 8 0.004 0.004 0.005 0.001 0.003
Rat 8 0.006 0.006 0.002 0.002 0.006