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Post Laboratory Discussion

EXPERIMENT NO.7
Experiment No. 7 to 12
Renato I. Dalmacio, RPh., MSPharm. Cholinergic Agonist &
Pharmacology 414 Antagonist
College of Pharmacy

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Pharmacologic Uses of
Cholinergic Agonist
Cholinergic Agonist
A. GIT
• These are substances that “mimic” the
effect of stimulation of the parasympathetic -Gastric Retention
nervous system. -Paralytic Ileus

B. GUT
-Urinary Retention

3 4

C. Eye E. Skeletal Muscle

-Glaucoma -Myasthenia Gravis
-Accomodative Esotropia

D. Cardiovascular Disorder F. CNS

-Supraventricular Tachycardia -Alzheimer’s Disease

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G. Dry Mouth
Cholinergic Antagonist
• These are drugs that oppose or nullify the
effect of stimulation of parasympathetic
H. Atropine Toxicity nervous system.

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Pharmacologic Uses of C. Respiratory

Cholinergic Antagonist -COPD
A. CNS -Asthma
-Parkinson`s Disease
-Motion Sickness D. Cardiovascular Disorder
-Myocardial Infarction
B. Opthalmologic Disorder
-Mydriasis
-Cycloplegia

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E. GI Disorders F. Urinary Disorder

-Peptic ulcer -Urinary Urgency

-Traveller’s Diarrhea -Bladder Spasm

-Hypermotility -Urinary Incontinence

-Enuresis

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G. Cholinergic Poisoning Mechanism of Action of

-Organophosphates Toxicity Pilocarpine
Acetylcholine released from the
H. Hyperhydrosis parasympathetic nerves activates
muscarinic receptors on effector cells to
I. Skeletal Muscle Relaxants alter organ function directly. Pilocapine
directly stimulates cholinergic receptors in
the eyes causing pupillary constriction,
J. Hypertension
spasm of accomodation and a transient
rise in intraocular pressure.
K. Smoking Cessation
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Adrenergic Drugs Atropine Poisoning

produces sympathetic nervous system Manifestations:
stimulation.
 Dry mouth

 Mydriasis
Cholinergic Drugs
produces parasympathetic nervous system  Tachycardia
stimulation.
 Hot & flushed skin

 Agitation
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 Delirium

Diuretics
• These are drugs that increase the flow of
urine.
EXPERIMENT NO.8
• It also inhibits sodium re-absorption to
Diuretic Property prevent edema.

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Diuretic Activity in Rats

(Lipschitz Test)
• This test is based on the principle that
SCREENING METHOD FOR water and sodium excretion in test
animals is different as compared to rats
DIURETICS treated with a high dose of urea.
Lipschitz Value
= urine excretion of test compound
urine excretion of urea treatment

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Diuretic Activity in Rats

Clinical Uses of Diuretic
(Actual Experiment)
% Increase in Urine Volume 1. Edematous State 2. Non-Edematous State
mL treated – mL negative control x 100 • Congestive Heart • Hypertension
Failure • Nephrolithiasis
mL negative control
• Kidney Disease • Hypercalcemia
Test animal
No.
Group Volume of urine (mL) TOTAL
(Volume of
MEAN
(Volume of • Hepatic Cirrhosis • Diabetes Insipidus
1st hour 2nd hour 3rd hour 4th hour 5th hour urine) urine)
• Idiopathic Edema
1 (-) control 0.6
0 0.1 0.15 0.1 0.2 0.55
2
0 0.1 0.15 0.15 0.2 0.6
3 (+) control 1.125
0.1 0.15 0.3 0.4 0.65 0.6
4
0.1 0.1 0.35 0.5 0.6 1.65
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Types of Diuretics
1. Carbonic Anhydrase Inhibitors
MOA: Carbonic anhydrase present in many
nephron sites catalyzes the dehydration of
H2CO3, a critical step in the reabsorption of
bicarbonate.
Site of Action: Proximal Convoluted Tubule

Examples:
Acetazolamide, Dichlorphenamide,
Methazolamide, Brinzolamide,
Tubule transport systems and sites of action of diuretics.
Dorzolamide,
ADH, antidiuretic hormone.
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2. Loop Diuretics 3. Thiazides

MOA: Inhibit the luminal Na+/K+/2Cl-
transporter in the thick ascending limb of
Henle’s loop.
Site of Action: Ascending limb of Loop of
Henle
MOA: Inhibit NaCl reabsorption from the
luminal side of epithelial cells in the distal
convulated tubule by blocking the
Na+/Cl- transporter.
Site of Action: Distal Convoluted Tubule

Examples: Examples:
Furosemide, Ethacrynic Acid, Hydrochlorothiazide, Indapamide,
Bumetanide, Torsemide Chlorthalidone, Metolazone,
Quinethazione
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4. Potassium Sparing Diuretics 5. Osmotic Diuretics

MOA: It reduce the Na+ reabsorption in MOA: It prevents the normal absorption of
the collecting tubules and ducts. water by interposing a countervailing
Site of Action: Late Distal Tubule and osmotic force.
cortical collecting tubule. Site of Action: Proximal Convoluted Tubule
and descending limb of Henle Loop.
Examples:
Spironolactone, Eplerenone, Examples:
Amiloride, Triamterene Mannitol, Glycerin,
Isosorbide, Urea
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Adverse Effects Associated to Diuretics

Types of Diuretics
Carbonic Anhydrase Inhibitors
Loop Diuretics
Thiazides
Potassium Sparing Diuretics
Osmotic Diuretics
Fasting of Test Animals
Adverse Drugs Reaction
Fasting of test animals is required prior to
biological test:
1. to increase the standardization of studies
on bioavailability and absorption
2. to ensure that accurate results will be
obtained from the test animals
3. to prevent interaction of foods to the
drug to be administered

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Fever
• It is a condition in which there is a rise in
body temperature above the normal that
EXPERIMENT NO. 10 is 37.5°C.

Antipyretic Activity • Fever may reflect infection or result from

tissue damage, inflammation, graft
rejection, or malignancy.

• Signs/Symptoms: shivering, headache,

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nausea, and constipation or diarrhea.

Antipyretic
• These are drugs that prevents or reduce
fever by lowering the body temperature. SCREENING METHODS FOR
ANTIPYRETICS
• The thermoregulatory center of the brain
is one likely to affected by these drugs.

• Examples:
Paracetamol, Aspirin, Ibuprofen
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Pyrogen Test, USP Brewer’s Yeast

The test involves measuring the rise in It was used to induce pyrexia (fever) by
temperature of rabbits following the administering 12.5-15.0% of this
intravenous injection of a test solution and suspension subcutaneously and regular
is designed for products that can be monitoring was recorder every hours prior
tolerated by the test rabbit in a dose not to administration of antipyretic agents.
to exceed 10 mL per kg injected
intravenously within a period of not more
than 10 minutes.

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% Protection
Other Test for Pyrogens
in Body Temperature
• Limulus Amoebocyte Lysate Test OR
=Initial Value –Final Value X 100
• Bacterial Endotoxin Test
Final Value
Test Group Temperature % protection
animal
No. Baseline 1st 2nd 3rd
hour hour hour
1
(-) control 40 39 38 37.5 4.80
2 41 40.5 39 38.5 4.24
3 (+) control
Paracetamol
44 41 38 36 14.78
4 (+) control
Aspirin 45 42 37 36 17.39
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Mechanism of Action Mechanism of Action

Aspirin Paracetamol
MOA: MOA:
Inhibit fever caused by agents that It is a weak COX-1 and COX-2 inhibitor
enhance the synthesis of IL-1 and other in peripheral tissues and possesses no
cytokines, which presumably cause fever, significant anti-inflammatory effects
at least in part, by inducing the
endogenous synthesis of prostaglandins.

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Clinical Uses Clinical Uses

1. Aspirin 2. Paracetamol
• Antipyretic • Antipyretic
• Analgesic • Analgesic
• Anti-Inflammatory • Preferred drug in patient allergic to aspirin
• Antiplatelet or when salicylates are poorly tolerated
• Ischemic attacks • It is preferable to aspirin in patient with
• Unstable Angina hemophilia or history of peptic ulcer and in
those whom bronchospasm is precipitated
• Eclampsia by aspirin
• Coronary Artery Thrombosis with Myocardial • It is preferred to aspirin in children with viral
Infarction infection.
• Thrombosis after Coronary Artery Bypass
Grafting 41 42

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Dosage & Administration Adverse Effects

1. Aspirin
Aspirin
• Gastric Upset
The antipyretic dose of salicylate for
• Gastric & Duodenal Ulcer
adults is 325 mg to 650 mg orally every 4
hours. • Hepatotoxicity
• Asthma
Paracetamol • Rashes
The conventional oral dose of • GI Bleeding
acetaminophen is 325 to 500 mg (650 mg • Renal Toxicity
rectally) four times daily.
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Adverse Effects
2. Paracetamol
• Mild increase in hepatic enzymes
• Dizziness
• Excitement EXPERIMENT NO. 11
• Disorientation
Analgesic Property

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Pain Analgesic
• It has been described by International
• Analgesics are agents that relieve pain by
Association for the study of Pain as an
elevating the pain threshold without
“unpleasant sensory and emotional
disturbing consciousness or altering
expressions associated with actual or
sensory modalities.
potential tissue damage” or described in
terms of such damage.

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Centrally Acting Analgesics

• Hot Plate Method
• Haffner’s Tail Clip Method
SCREENING METHODS FOR • Tail Immersion Test
ANALGESICS • Radiant Heat Method
• Formalin Test in Rats
• Tooth Pulp Stimulation
• Grid Shock Test
• Electrical Stimulation of the Tail
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Peripherally Acting Analgesics % Analgesia

• Pain in inflamed tissue 1 - # of writhes of individual mouse X 100
-a.k.a.”Randall-Selitto Test” Mean number of writhes of (-) control group

Test Group # of MEAN % Analgesia

animal writhes
• Writhing Test No.
1 36 X
(-) control 37
2 38 X
3 10 X 72.97%
(+) control
4 Aspirin 8 X 78.38%

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Narcotic Analgesics Non-Narcotic Analgesics

• These are potent analgesics which are • A drug which relieves pain, but which does
effective for the relief of severe pain. not have narcotic effects.
• It relieve pain by acting directly on the • It have principally analgesic, antipyretic,
central nervous system. and anti-inflammatory actions.
• They bind to opioid receptors, which • They do not bind to opioid receptors and
present in many regions of the nervous are not classified under the Controlled
system and are involved in pain signalling Substances.
and control. • Examples:
• Examples: Aspirin, Ibuprofen, Acetaminophen
Morphine, Methadone, Codeine,
Hydrocodone, Oxycodone 53 54

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Narcotic Non-Narcotic Advantages of NSAID’S

Analgesics Analgesics
Act centrally • Are analgesic and also reduce inflammation.
Cause addiction • Are sufficient to manage many forms of pain.
• Can be combined with Opioids.
Produce CNS • They do not posses large risk of addiction,
depression dependence, tolerance or withdrawal.
Do not produce • It has higher margin of safety that can be
gastric irritation given in higher doses.
Has no anti- • Most of these drugs are well absorbed, and
food does not substantially change their
inflammatory effect bioavailability.
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Mechanism of Action
Inhibits pain stimuli at subcortical site
by irreversibly inhibiting the COX pathway
in the CNS.
EXPERIMENT NO.12

Hypoglycemic Property

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Classical Symptoms of Diabetes

Diabetes Mellitus
Mellitus
• It is defined as an elevated blood glucose
associated with absent or inadequate • Polyuria
pancreatic insulin secretion, with or
without concurrent impairment of insulin • Polydipsia
action.
• Polyphagia

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Models for IDDM

• Alloxan-Induced Diabetes
SCREENING METHODS FOR • Streptozocin-Induced Diabetes
HYPOGLYCEMICS • Virus-Induced Diabetes
• Insulin Antibodies-Induced Diabetes
• Hormone-Induced Diabetes

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Models for Insulin Sensitivity and

Models for NIDDM
Insulin-Like Activity
• Streptozocin-Induced Neonatal Model for • Euglycemic Clamp Technique
NIDDM
• Adrenaline-Induced Acute Hyperglycemia • Assay for Insulin and Insulin-Like Activity
• Dithizone-Induced Diabetes

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% Reduction in Blood Sugar Level % Reduction in Blood Sugar Level

Initial glucose level(0 min) - Succeeding glucose levelX 100 Initial glucose level(0 min) - Succeeding glucose levelX 100
Initial glucose level (0 min) Initial glucose level (0 min)
Test Group Blood Sugar Levels ( mg/dL)
Animal
No. Test animal No. Group Percent (%) Blood sugar reduction
FBS after 0 15 30 60 15 minutes 30 minutes 60 minutes after
glucose minute minutes minutes minutes after treatment after treatment treatment
administra after after after after
tion treatmen treatmen treatmen treatmen 1
(-) 3.45 6.90 13.79
t t t t
2 control
1 80 145 145 140 135 125 3.33 6.67 13.33
(-) control
2 75 150 150 145 140 130 3
(+) 9.09 18.18 33.33
3 85 165 165 150 135 110 4 control
(+) control 8.82 23.53 32.35
4 90 170 170 155 130 115

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Four Categories of Diabetes Mellitus :
Type II Diabetic Mellitus:
Characterized by tissue resistance to
the action of insulin combined with a
relative deficiency in insulin secretion.
Formerly known as” Non-insulin
Dependent DM”.
Dehydration in untreated and poorly
controlled individuals with type 2
diabetes can lead to a life-threatening
condition called Nonketotic
Hyperosmolar Coma. 69
Four Categories of Diabetes Mellitus :
Type IV Diabetes Mellitus:
a.k.a “Gestational diabetes(GDM)”.
Defined as any abnormality in
glucose levels during pregnancy.
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Four Categories of Diabetes Mellitus :
Type I Diabetes Mellitus:
Selective beta cell (B cell) destruction
and severe or absolute insulin
deficiency.
Formerly known as “Insulin-
Dependent DM”.
Interruption of the insulin replacement
therapy can be life-threatening and can
result in Diabetic Ketoacidosis (DKA) 68
Four Categories of Diabetes Mellitus :
Type III Diabetes Mellitus:
Refers to multiple other specific
causes of an elevated blood
glucose.
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Diagnosis:
Diabetes mellitus is characterized by
recurrent or persistent hyperglycemia, and is
diagnosed by demonstrating any one of the
following:
• FPG ≥ 7.0 mmol/L (130 mg/dL)
• RPG ≥ 11.1 mmol/L (200 mg/dL)
• Hb A1C ≥ 6.5%.
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DRUGS THAT MAY INCREASE BLOOD DRUGS THAT MAY DECREASE BLOOD
SUGAR LEVEL SUGAR LEVEL
• Corticosteroids • Phenothiazines • Acetaminophen
• Estrogens • Phenytoin • Alcohol
• Glucagon • Salicylates • Anabolic Steroids
• Isoniazid • Thiazide diuretics • Clofibrate
• Lithium • Triamterene • Disopyramide
• Oral Contraceptives • Tricyclic • Gemfibrozil
Antidepressants • Monoamine Oxidase Inhibitors (MAOIs)
• Pentamidine
• Beta-Blockers
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Types of Insulin Oral Hypoglycemic Agents

1. Rapid Acting Insulin 1. Insulin Secretagogues

-Insulin Lispro, Insulin Glulisine & Insulin Aspart a. Sulfonylureas
2. Short Acting Insulin 1. First Generation
-Regular Insulin -Tolbutamide, Chlorpropamide, Tolazamide
3. Intermediate Acting Insulin 2. Second Generation
-Isophane Insulin & Lente Insulin -Glyburide, Glipizide, Glimepiride
4. Long Acting Insulin b. Meglitinides
-Repaglinide
-Ultralente Insulin, Insulin Glargine, Insulin
Detemir c. D-Phenylalanine Derivative
75 -Nateglinide 76

2. Insulin Sensitizers MISCELLANEOUS

a. Biguanides
-Metformin • Pramlintide
b. Thiazolidinediones
-Pioglitazone • Exenatide
-Rosiglitazone
3. α-Glucosidase Inhibitors • Sitagliptin
-Miglitol
-Voglibose
-Acarbose
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