SCREENING OF

ANTIDIABETIC
DRUGS
PRESENTED BY,
SHARON SEQUEIRA
1ST M.PHARM
DEPARTMENT OF
PHARMACOLOGY
 INTRODUCTION
 Diabetes mellitus is a disease characterized by
persistent hyperglycemia , resulting either from
inadequate secretion of the hormone insulin, an
inadequate response of target cells to insulin
combination of these factors.
 Type 1 diabetes mellitus is characterized by loss of
the insulin-producing beta cells islets of Langerhans
of the pancreas.
 Type 2 diabetes mellitus is a metabolic disorder that
is primarily characterized by insulin resistance,
relative insulin deficiency, and hyperglycemia.
 Gestational diabetes, Type 3 ,also involve a
combination of inadequate insulin secretion
responsiveness , resembling type 2 diabetes in
several respects .
 It develops during pregnancy and improve or
disappear after delivery.
 Normal blood glucose level-120mg/dl.
 Fasting blood glucose level-100mg/dl.
 PATHOLOGICAL CHANGES
 Increase in vessel wall matrix.
 Thickening of capillary basement membrane.
 Cellular proliferation resulting in vascular
complication:

 Lumen narrowing
 Early atherosclerosis
 Retinopathy
 Neuropathy
 SYMPTOMS OF DIABETES
 Polyuria [increase urination]
 Polydipsia [increase thirst]
 Polyphagia [increase hunger]
 TREATMENT
 Conventional preparations of insulin
o Short acting:
 Regular insulin
 Prompt insulin Zinc suspension (semi lente)
 Intermediate Acting
 Insulin Zinc suspension(Lente)
 Neutral protamine hagedron or Isophane insulin
 Long Acting
 Extended insulin Zinc suspension (Ultra lente)
 Protamine Zinc insulin.
Purified Insulin Preparations
 Single Peak Insulins :Actrapid, Lentard,
Actraphane
 Monocomponent Insulins :Monotard
 Human Insulin :Human Actrapid.
Oral Hypoglycemics
1.Sulponyl Ureas
a.First Generation
 Tolbutamide
 Chlorpropamide
 Acetohexamide
b.Second Generation
 Gilbenclamide
 Glipizide
 Gliclazid
2.Biguanides
 Phenformin
 Metformin

3.Miscellaneous
 Acarbose
 Guargum
VARIOUS SCREENING MODELS
 IN VIVO METHODS
1.Alloxan induced diabetes.
2.Streptozotocin induced diabetes.
3.Hormone induced diabetes
a.Growth hormone induced diabetes
b.corticosteroid induced diabetes [DEXAMETHASONE]
4.Insulin deficiency due to insulin antibodies
5.Surgically induced diabetes
1.ALLOXAN INDUCED DIABETES
 Purpose
 Induction of diabetes using chemical alloxan.
 Alloxan is toxic glucose analouge which selectively
destroys insulin producing beta cells when
administered to species
 IN most species , triphasic time course is observed
.[Initial rise of glucose is followed by sustain
increase of blood glucose].
Animal used
 Baboons 65-200mg/kg
 Beagle dogs 60mg/kg
 Wistar rat 100-
175mg/kg
 Rabbits 150mg/kg
 PROCEDURE

Animal is injected with a single dose [100mg/kg body

weight] dissolved in normal saline by i.p route.

Animals are kept for 48 hours during which food and

water is allowed add libitum.
 Blood glucose levels show triphasic response with
hyperglycemia for 1 hour followed by hypoglycaemia that lasts
for 6 hours and stable hyperglycemia after 48 hours.

Animals showing fasting blood glucose level above

140mg/kg after 48 hours are considered diabetic.

For a period of six weeks , drug samples to be screened

are administered orally.
 After six weeks of treatment , blood samples are
collected from 8 hour fasting animals [can be
collected via orbita sinus through a pipette].

The serum glucose level is estimated by glucose oxidase-

peroxidase method[GOD-POD kit] using autoanalyser.
 EVALUATION
 Blood glucose level is estimated.
 Compare the result of test with standard and control.
 DRAWBACKS
 High mortality
 Ketosis
 Diabetes induced is reversible.
 Cannot be done on guinea pigs.
2.HORMONE INDUCED DIABETES
 Principle:
Dexamethasone is a long acting glucocorticoid possessing immuno suppressant
action in the islets and produces type -1 diabetes.
i.E they stimulate glucogenesis in liver resulting in increase hepatic glucose output
And insulin resistance and causes hyperglycemia.

Adult rats 150-200gm dexamethasone 2-

PROCEDURE: 5mg/kg i.p

Repeated injection of same dose level is

carried out for a period of 20-30 days
resulting in IDDM.
 The sample to be screened is administered
through a suitable route , blood glucose is
measured.

 Limitations:
 Long standing procedure.
3.INSULIN DEFICIENCY DUE TO INSULIN
ANTIBODIES
 PRINCIPLE
 A transient diabetic syndrome can be induced by
injecting guinea pigs anti insulin serum.
 Diabetes persists as long as antibodies are capable of
reacting with insulin remaining in the circulation.
 Preparation of antibody:

 Bovine insulin, dissolved in acidified water [PH- 3.0]

at a dose of 1mg is injected to guinea pigs weighing
300-400gm.Anti insulin sera is collected after two
weeks of antigenic challenge.
PROCEDURE:

Adult albino rat are injected with 0.25-1.0 ml
of guinea pig anti-insulin serum.

Insulin antibodies induce a dose dependent

increase of blood glucose level up to 300mg/dl.

The drug sample to be screened is administered by a

suitable route and blood glucose level is analysed to
determine the activity.
 LIMITATIONS
 Effect persist as long as antibodies remain in the
circulation.
 Large doses and prolonged administration
ketonaemia, ketouria.
4.STREPTOZOTOCIN INDUCED DIABETES
 Streptozotocin (STZ and Zanosar) is a naturally
occurring chemical that is particularly toxic to the
insulin producing beta cells of the pancreas in
mammals.
 It is used in medicine for treating certain cancers
of the Islets of Langerhans and used in medical
research to produce an animal model for diabetes.
 Mechanism of causing beta cell
damage
 By process of methylation.
 Free radical generation.
 Nitric oxide production.
 PROCEDURE:
 STZ induces diabetes in almost all species of animals.
 Diabetogenic dose varies with species and the optimal
doses required in various species are in rat (50-60mg/kg
,i.p or i.v.) , mice (175-200mg/kg, i.p. or i.v) and dogs (15
mg/kg, for 3 days).
 The blood glucose level shows the same triphasic response
as seen in the alloxan treated animals , hyperglycemia at
1 hr, followed by hypoglycaemia , which last for 6 hr and
stable hyperglycemia 24-48 hr after STZ administration.
 ADVANTAGES
1. Greater selectivity towards beta cells .
2. Low mortality rate.
3. Long and irreversible diabetes.
5.SURGICALLY INDUCED DIABETES
It can be done by removing the pancreas as the whole or a part of
it. In partial pancreactomy more than 90% of the organ must be
removed to produce diabetes.

Depending on the amount of intact pancreatic cells, diabetes

may range in duration from a few days to several months.

Total removal of pancreas results in an insulin dependent

form of diabetes mellitus and insulin therapy is required to
maintain experimental animals.
The portion of the pancreas usually left
intact following a subtotal pancreatic
resction is typically the anterior lobe.
 ADVANTAGES
 Avoids cytotoxic effects of chemical diabetogens on other
body organs.
 Resembles human type-2 diabetes due to reduced islet
beta cell mass.

DISADVANTAGES

 Post operative procedures.

 Digestive problems due to excision of exocrine portion of
the pancreas.
 Loss of counter regulatory response to hypoglycaemia.
 High mortality.
 INVITRO METHODS
 ISOLATED PANCREAS OF RATS

 PRINCIPLE

To test the effect of test compound on the hormones

(insulin , glucagon and somatostatin ] in response to
elevated glucose level.

ANIMAL USED:
Male Wistar Rats(200-250g).
 PROCEDURE
After anaesthesia , pancreas are
removed.

Perfused with Kreb –Ringer

bicarbonate buffer , with albumin and
glucose.

Perfusate is collected every minute

for 30 mints.
Test compound is added from 5th min to 15th
mints.

Glucose is added from 16th mints to 30th mints.

Insulin, Glucagon, somatostatin are estimated

radioimmunology.

The effects of test compound whether it increases or

decreases the secreted hormones of pancreas in
response to elevated glucose level is compared with
the control
 REFERENCE

 Essential of medical pharmacology –K.D.Thripati

 Drug discovery and clinical research –S.K Guptha
THANK YOU