“SCREENING MODELS OF ANTI-ULCER AGENT”

PREPARD BY :-

Shree S.K. Patel College Of Pharmaceutical Mitesh Patel

Education And Research (20022411072)
Ganpat University , Ganpat Vidhyanagar - 384012 ,
Dist – Mehsana , Gujarat , India.

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Definition
Peptic ulcer is a chronic inflammatory condition
in which hole or open sore in the lining of the
stomach, duodenum or oesophagus.

The most common etiologies of gastric ulcers

are Helicobactor pylori infection and NSAID
use.

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Pathophysiology

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 SCREENING MODEL
 In-vivo models
(1) Pylorus ligation in rats (SHAY rat) model
(2) Stress ulcer through immobilization stress
(3) Stress ulcer by cold water immersion
(4) Histamine induced gastric ulcer
(5) NSAID induced ulcer in rats (Indomethacin, Aspirin)
(6) Ethanol induced mucosal damage in rats
(7) Acetic acid induced gastric ulcer
(8) Subacute gastric ulcer in rats
(9) Gastric ischemia reperfusion injury in rats

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 In-vitro models
(1) [I¹²⁵] Gastrin binding assay

(2) Tiotidine binding assay

(3) H+/K+ ATPase inhibiting assay

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(1) HISTAMINE INDUCED GASTRIC ULCER
 Principle :-
• When histamine is administered intraperitoneally,
• Histamine binds with receptors present on the surface of
parietal cells,
• Activation of Adenyl Cyclase,
• So convert ATP into C-AMP,
• This conversion enhance secretion of HCL from parietal cells.

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 Procedure :-
• Take 300-400 gm weighted guinea pig
• Animals are fasted for 36 hr before Experiment
• 1 ml Histamine acid phosphate was administered Intraperitoneal to induce ulcer
• To Prevent Histamine toxicity, Promethazine Hydrochloride in dose of 50 mg is
injected I.P., 15 min before and 15 min after histamine injection.
• The Standard / Test drug are administered Orally or S.C. before 45 min of
Histamine injection.
• After 4 hr, animals are sacrified, Stomach removed
• Stomach is opened along the greater curvature
• Identify ulcer scoring to Examine the severity of ulcers

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 Evaluation :-
Type 0 : No visible ulcers
Type 1 : 10 or less ulcers, 1-3 mm in diameter
Type 2 : 11 or more ulcers, 1-3 mm in diameter
Type 3 : 1 or more ulcers, 4-6 mm in diameter
Type 4 : 1 or more ulcers, 7 mm or more in diameter
Type 5 : Perforation of the gastric wall

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(2) STRESS ULCER THROUGH IMMOBILIZATION
STRESS

 Principle :-
• Psychogenic factors, such as stress, play a major role in the
pathogenesis of gastric ulcers in man.
• Hanson and Brodie (1960) described methods to study the effect
of anti-ulcer drugs on immobilization stress in rats.

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 Procedure :-
• Groups of 10 female Wistar rats weighing 150–170 gm are used.
• Food and water are withdrawn 24 hr before the experiment.
• After oral or subcutaneous administration of the test compound or the placebo
solution (No therapeutic benefit) the animals are slightly anesthetized with
ether.
• Both lower and upper extremities are fixed together and the animals are
wrapped in wire gaze.
• They are horizontally suspended in the dark at 20 °C for 24 h and finally
sacrificed in CO2 anesthesia.
• The stomach is removed, fixed on a cork plate and the number and severity of
ulcers is registered with a stereo-microscope.

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 Evaluation :-

• Ulcer score 0 = no ulcer

• Ulcer score 1 = superficial ulcers
• Ulcer score 2 = deep ulcers
• Ulcer score 3 = perforation

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(3) NSAIDS INDUCED GASTRIC ULCER MODEL
 Principle :-
• Prostaglandins play a protective role via stimulating the secretion
of mucous, maintaining blood flow and regulating mucous cell
turn over and repair.
• NSAIDs cause ulcers by inhibiting prostaglandins synthesis by
inhibiting cyclooxygenase enzyme in COX pathway.

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 Procedure :-
• Select albino rats weighing 200-300 gm and divide them into two
groups consisting of 3 animals in each group.
• Group-1 (control group) receives water or 1% CMC.
• Group-2 (Test drug treated group)
• Animals are fasted for 24-36 hours

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• In both group, orally administer NSAID (aspirin or indomethacin)
using appropriate vehicle (water or 1% carboxymethylcellulose)
• After 1 hour, Group-2 animals are treated with test drug.
• 4 hours later, animals are sacrificed, stomach is removed and severity
of ulcer is measured.
• Dose of NSAIDs used to induce ulcers :-
(1) Aspirin-150mg/kg of body weight
(2) Indomethacin-100mg/kg of body weight.

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Evaluation :-

 Intensity of ulcers with scoring :-

• O - normal coloration
• 0.5 - red coloration
• 1 - spot ulcer
• 1.5 - hemorrhagic streaks (bands)
• 2 - deep ulcer (ulcer count ≥ 3 but ≤ 5)
• 3 - perforations (ulcer count > 5)

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(4) [I¹²⁵] GASTRIN BINDING ASSAY

 Principle :-
• Gastrin (G cell of gastric antrum) is major stimuli for gastric acid secretion.
• Gastrin binds to cholecystokinin (CCK2) receptor on basal membrane of
Perietal cells to induce expression of H+/K+ ATPase pump (Proton Pump).
• Increased activity of H+/K+ ATPase result in increased secretion of gastric
acid into Stomach.
• Compounds with gastric receptor antagonistic activity can Prove to be
useful as anti-ulcer drugs.

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 Procedure :-
• The assay is done by Fundic gland Suspension Obtained from guinea pig
stomach for binding assay
• The fundic gland suspension incubated with 50 μL [I¹²⁵] Gastrin for 90 min
at 37°c temperature.
(1) In Presence of buffer alone (For total binding)
(2) In Presence of unlabeled gastrin (For non-specific binding)
• Now, In micro-centrifuge tube, ice cold buffer is layered with Incubated
mixture and centrifuged for 5 min at 10,000 rpm.
• Radioactivity is quantified in pellet after discarding the supernatant.

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 Evaluation :-

• Total binding, Non-specific binding,

Specific binding (Total binding - Nonspecific binding) calculated.
• Percentage specific bound [I¹²⁵] gastrin displaced by a given
concentration of the test compound calculated.
• The higher the displaced [I¹²⁵] gastrin, More is the gastrin
antagonistic activity of test compound.

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