1. Anti-inflammatory Drugs
# Lab 4 #

2. Permanent destruction
Inflammation:
it is a biological response of vascular tissues to harmful stimuli, such as pathogens, damaged cells, or irritants.
Inflammation may ends with either :
Complete healing
of tissues
Permanent destruction
of tissues

3. Signs of inflammation Redness: Hotness:
Due to vasodilatation by effects of releasing of histamine, bradykinin and prostaglandin.
Hotness:
Due to increased blood flow.

4. Signs of inflammation Swelling: Pain:
due to increased vascular permeability by the released mediators and increased the exudate in the inflammed area
Pain:
due to irritation of nerve ending by inflammation and the pressure of the swelling on the nerve ending.

5. Inflammatory mediators:
histamine
5-HT (serotonin)
Bradykinin
Prostaglandins (eg PGE2 )
Interleukines
Substance P
Nitrous oxide
Main
inflammatory
mediator

6. Phases of Inflammation
Fluid phase (vascular phase):
Increased vasodilatation leads to increased permeability of the vascular bed to plasma protein.
- Increasing fluid will help in:
1- dilution of the irritant
2- increase conc. of antibodies from blood to inflamed area
3- supply nutrients to the immune cells.

7. Phases of Inflammation
Cellular phase (exudative phase):
Involves migration of tissue macrophages and polymorphonuclear leukocytes (PMNL) to the inflamed area.
Fibrous phase (proliferative phase):
A new connective tissue (fibrous) containing fibroblast and capillaries is formed.

8. Classification of the Inflammation
Non – immunological :
Induced by chemical irritants such as formalin
Immunological :
Induced by infections such as bacterial infection

9. Anti-inflammatory Drugs
Steroidal
Non-steroidal
- Cortisone
- Hydrocortisone
- Acetaminophen
- Aspirin

10. Steroids (SAIDs) - Containing steroid moiety in their sturcure
Glucocorticoids (GC)
Cortisone

11. Glucocorticoids (GC) Synthetic Natural - Cortisone - Betamethasone
- Hydrocortisone
- Betamethasone
Dexamethasone
- Predinsone
Fluorinated
Glucocorticoids
Liver enzymes
Prednisolone

12. Glucocorticoids (GC) Mechanism of Action : -
They act by indirect inhibition of the enzyme phospholipase A2
which activate synthesis of arachidonic acid with subsequent
formation of prostaglandins.
They induce synthesis of a protein “lipocortin-1” which has
the inhibitory effect on phospholipase A2. -

13. GC inhibition
phospholipase A2

14. Side Effects : Immunosuppression
Hyperglycemia due to increased gluconeogensis, insulin resistance, and
impaired glucose tolerance ("steroid diabetes");
Steroid-induced osteoporosis: reduced bone density (osteoporosis,
Osteoporosis , higher fracture risk, slower fracture repair)
Redistribution of body fat: moon face, buffalo hump and truncal obesity.
Adrenal insufficiency
Muscle breakdown (proteolysis), weakness; reduced muscle mass and repair
Anovulation, irregularity of menstrual periods
Growth failure, pubertal delay
Increased plasma amino acids, increased urea formation;
Glaucoma due to increased cranial pressure

15. Side Effects :
Moon face
buffalo hump

16. Non-Steroidal Anti-inflammatory Drugs
(NASID)
They don’t contain steroid moiety
They also have analgesic and antipyretic activity

17. Mechanism of Action :
NSAIDs inhibit synthesis of PGs which are the main factors
Playing a role in the inflammaltion.
Inhibit synthesis of PGs through inhibition of cyclooxygenase
Enzymes which are responsible for production of PGs

18. GC inhibition
phospholipase A2

19. Cyclooxygenese ( COX ) Isoforms :
- Constitutive
- Many tissues ( blood vessels
stomach and kidney )
- inducible
By inflammatory processes
and mediators
COX 3
has recently been described

20. Non-Steroidal Anti-inflammatory Drugs
(NASID)
Non-selective
COX inhibitors
selective COX2
inhibitors
- Aspirin
- Ibuprofen
- Diclofenac
- Meloxicam
- Celecoxib
- Rofecoxib

21. Side Effects : Unwanted effects, owing largely to inhibtion of COX1
Particularly in the elderly and include :
- Despepsia, nausea and vomiting , ulceration and gastric
damage in chronic users, with risk of hemorrhage
- Reversible renal insufficiency
- Analgesic-associated nephropathy ( irreversible )
- Liver disorders, in high doses, e.g. acetaminophen

22. Measurement the activity of anti-inflammatory drugs
- Method : Paw Oedema Method
Principle : induction a chemical inflammation
by injecting an irritant ( formalin ) into
rat’s paw
Objective : measure the anti-inflammatory activity of
diclofenac and hydrocortisone with
different doses )

23. Procedure : 1- select 5 rats
2- inject each rat 1 ml urethane for anesthesia.
3- select one as control and inject the rest of them intraperitoneal
rat 1 >>> control
rat 2 >>> 40 mg/kg diclofenac
rat 3 >>> 80 mg/kg diclofenac
rat 4 >>> 20 mg/kg hydrocortisone
rat 5 >>> 40 mg/kg hydrocortisone
4- after 1 hr , inject 0.1 ml formalin in each rat ( 2 to 5 ) into
their paws >>> to induce inflammation.
5- after 1 hr , take the reading using the plythysmometer of each
rat paw ( right and left ).
6- calculate the inflammation and response % for each drug.

24. Response % = ــــــــــــــــــــــــــــــــــــــــ X 100 C
inflammation LP RP
Dose
___ C
control T1
40 mg/kg vol. T2
80 mg/kg vol. T3
20 mg/kg hydro. T4
40 mg/kg hydro.
Inflammation = RP - LP
C - T
Response % = ــــــــــــــــــــــــــــــــــــــــ X 100 C
Response % >>>> Anti-inflammatory activity