Animal models can In “Vitro” – glass tests are experiments carried out in the

laboratories using test tubes, beakers

be used to predict
the toxicity, In “Vivo” (life) are experiments conducted in animal models
safety and efficacy
of various
compounds in
clinical research
 “Any programme or project involving use of animal(s) for the
acquisition of knowledge of a biological, physiological,
ethological, physical or chemical nature; and includes the use of

Experiment animals in the production of reagents and products such as

antigens and antibodies, routine diagnostics testing activity and
definition establishment of transgenic stocks for the purpose of saving or
prolonging life or alleviating suffering or significant gains in the
well being for people of the country or for combating any disease,
whether of human beings, animals or plants”
The Drug Discovery
Drug Discovery: The identification of a
molecule that could potentially lead to the
development of a new medicinal product.

Target identification and validation: The

identification of the target that the new drug is to
act against.

Hit identification and validation: The

identification of molecules that will act against the
target, known as ‘hits’.

Hit to Lead (lead generation): analysis of the

drug’s properties, such as potency, solubility,
selectivity, and stability. In vitro and in
vivo studies (using animal models)

Lead Optimization: The characterization of the

drug and the study of its properties. Again, in
vitro, and in vivo testing can be adopted.
 Very limited number of
studies can only be done on
humans

Physiology / anatomy can be

matched to humans

Animals are mostly

Why susceptible to the diseases
that affect humans
animals are Short life span allows to be
used ? studied through their entire
life in a limited period

Controlled experiments and

environmental variables can
be minimized.

The dosage/route/of exposures

can be controlled/varied and
experiments can be replicated
 1. Used in several areas of bio medical research and product
testing
2. Developing new treatments for diseases or ways of preventing
diseases

Scope 3.
4.
Fundamental biological and medical research
Safety testing of non medical products used in thee
households
5. Agriculture and industry
6. Developing new methods of diagnosis
 The drug/ substance to be tested for the first tie in the animals should be for
specific purpose

The detailed literature review and ccomparison of data

What all Computational modelling, simulation r insilico analysis

should be done In vitro assays with cell line/ tissue culture should support the desired
before activity

Ex vivo studies with organs collected from slaughter houses or culling

animals may be used

Obtain the approval of IAEC or CCSEA

Which are the
animals subject
to regulations  Most of the animals comes under CPCSEA
 Anything higher than invertebrates needs regulation

 Invertebrates (e.g.; cockroaches)< Birds <rodents <canines/felines

<bovines/equines <primates(e.g. Rhesus macaque) <more evolved
primates (e.g. Chimpanzee)
 Two types

Types of
animal models
Homology (structures
Analogy (similar derived from the same
structures imply similar evolutionary precursor
functions) have the same or similar
functions).
 There are four types
1. Induced or experimental models, that attempt to
reproduce conditions found in the original species
Categories of 2. Spontaneous or natural models, that are recognized as
animal being similar to some condition in the original species

models 3. Negative or nonreactive models, that are the normal

counterparts of a disease model,
4. Orphan models, that are animal diseases for which no
human or animal counterpart is known.
 1. Appropriateness as an analog
2. Transferability of information
3. Genetic uniformity of organism, where applicable
The selection 4. Background knowledge of biological properties
of any model 5. Cost and availability,
6. Generalizability of the results
7. Ease of and adaptability to experimental manipulation
8. Ecological consequences,
9. Ethical implications.
 Cost-effective
 Cardiovascular disease
Rodents

Rodents (mainly mice and rats)  Cancer
Breed fast
have been used in medical  Alzheimer’s disease
research for many years due to  Parkinson’s disease
their similarities to the human Require
genome.  Cystic fibrosis minimal living
The Foundation for Biomedical  Transplants space,
Research states that 95% of  Respiratory problems
animal models for research are Are easy to
mice and rats.  Cataracts
care for
Most human disease genes exist in  Diabetes
mice and rats, making them  Hypertension
suitable for research.
 Seizures
 Substance addiction (for anti-craving)
 Behavior studies (rats have a larger brain than mice and are
frequently used in psychological research)
 Zebrafish are used in 80% of medical research projects and are the main
non-mammal in research.
These tiny Asian freshwater fish hold 80% of the human genome code
and are becoming increasingly popular for use in research.

Zebrafish  They have many similar organs to humans; two eyes, a heart, a brain,
a pancreas, a liver, and kidneys.
(Danio rerio)  A zebrafish lays several hundred eggs at one time (which can be
weekly)
 Zebrafish development is fast
 Cardiovascular disease (a zebrafish can regenerate its heart)
 Metabolic diseases
 Cancer
 Neurological disorders and Motor behavior
 Tissue regeneration after injury
 Acute toxicity study: Adverse effects occurring within a short time
following administration of single dose.

Oral toxicity Sub- acute toxicity- 4 to 8 weeks

studies

Chronic toxicity- Period of 90

days or more.

Carcinogenicity

Toxicological Special toxicity

Mutagenicity

Investigation studies- Period of 2

years or more
Teratogenicity

Reproductive toxicity and

development toxicity tests

1. Acute dermal
toxicity
Dermal toxicity
studies
2. Repeated dose
dermal toxicity
  It refers to recording of adverse signs and symptoms
after the administration of single dose of drug at several
levels higher than the therapeutically equivalent dose.
 Procedure:
• Following OECD guideline 425 with 2000mg/kg as
limit test.
Acute oral • Five animals in each group is treated with 2000mg/kg
and observed for 14 days.
toxicity  Examination
Gross behaviour and mortality, if any is observed
throughout the study period for 14 days.
All the animals should be observed at ½, 1, 2, 3, 4, 5,
6, 24 hours after treatment and daily once a time for
any mortality during the entire period of the study (14
days).
 OECD 408
(90 day oral repeated dose toxicity study)/AYUSH 170 guideline [Guidelines
for pre-clinical safety evaluation for Ayurveda, Siddha and Unani drugs and
other traditional medicines in India] should be adopted.
Parameters:
Ponderal change: Body weight, organ weight
Chronic oral toxicity Hematological parameters:
Total WBC count, differential count (Neutrophils, Lymphocytes,
Eosinophils, Monocytes, Basophils), haemoglobin, PCV, ESR, RBC
count, platelet count, MCV, MCH and MCHC.
Biochemical parameters:
Serum urea, creatinine, total protein, albumin, globulin, alkaline
phosphatase, S.G.O.T., S.G.P.T., total bilirubin, direct bilirubin, uric acid
and serum calcium
Histopathology
 OECD 407
(28 day oral repeated dose toxicity study in rodents)
Sub-chronic oral Ponderal change: Body weight, organ weight.
toxicity Serum parameters
Hematological parameters
Histopathology
Reproductive toxicity study is a requirement for any new drug
compound before administration to any female of child bearing age.
The studies are used to determine any effect of active ingredients on
the reproductive system.
This study may be divided into three segments:
 Segment I reproductive toxicity study documents effects of the drug
on fertility.
 Segment II and III documents such effects as on the embryo, pre
and post natal developments.
Multigenerational effects and teratology are also studied
  These tests are aimed at ruling out mutagenic
effects of a new drug compound
 Any effects on the chromosomes or the DNA
Genetic Toxicity strands may be a precursor for gene mutation.
 Salmonella typhimurium is usually used in
assays to detect mutation
  Testing for carcinogenicity usually applies to drugs for treatment of
chronic disease conditions.
 The duration of this test is about 18 – 24 months.
 The surviving animals are killed and studied at specified weeks
Carcinogenicity during the period of the study.
Studies  Data collected are
 animal deaths,
 Tumour incidence
 Type and site of tumour appearance (if any)
 Pre-neoplastic tissue changes etc
 Replacement
This advocates use of alternative methods.

The principle
of Three Rs’
Reduction
This principle advocates the application of
methods that employ less number of animals

Refinement
This preaches the principle that methods
adopted should be such that pain, suffering and
distress is minimized for the animals that are
still in use in the preclinical research process. It
encourages animal welfare and care.
4th R
Reuse and
rehabilitation (2004)
  The prevention of cruelty to Animal Act (PCA) 1960
 CPSSA regulated by PCA
 CPSSA formerly called as CPCSEA

Animal  “To take all such measures as may be necessary to ensure that animals are not
subject to unnecessary pain or suffering before, during or after the
experimentati performance of experiments on them”
 Breeding of and experiments on animals (control & supervision) rules 1998
on in India
 Experiments using animals are conducted as per OECD guidelines
 Experiments using animals are reported using ARRIVE guidelines
 What is CPCSEA stand for?
 The Committee for the Purpose of Control and Supervision of
Experiments on Animals (CPCSEA)
 Committee for Control and Supervision of Experiments on
Animals (CCSEA)
Functions of
CCSEA
 Animals lowest on the
phylogenetic scale which may
give scientifically valid results
should be used for any
experimental procedure Experiments on animals may be
carried out for the purpose of
Proper use of animals in
advancement by new discovery
experiments and avoidance or
minimization (when avoidance is
of physiological knowledge,
not possible)
knowledge which is expected to
5 principles of be useful for saving or
CCSEA prolonging human life or
Persons engaged in animal
experimentation have a moral
responsibility for the welfare of
alleviating suffering
the animals after their use

Living conditions of the animals

should be appropriate for their
species and contribute to their
health and comfort
 Use of stray dogs for new vaccine trials and challenge
studies of vaccines subject to adhering to its guidelines.
Recent The move has resulted in protest from animal rights
organisations like the People for Ethical Treatment of
recommendation Animals (PETA), India, which alleged that the decision
will open floodgates not only for stray dogs, but also other
feral animals to be used in experiments.
OECD
The OECD
Guidelines set
standards for The Organization for Economic
responsible business Cooperation and Development
conduct across a
range of issues such Part I defines standards for
as human rights, responsible business conduct in
areas such as labour rights, human
labour rights, and the rights, environment, information
Part II sets out Procedural
Guidance to implement the
environment disclosure, combating bribery,
recommendations in Part I.
consumer interests, competition,
taxation, and intellectual property
rights.
 402 Acute dermal Toxicity
404 Acute dermal Irritation/ Corrosion
403 Acute inhalation Toxicity
405 Acute Eye Irritation /Corrosion
406 Skin sensitization
Important OECD
guideline for toxicity 407 28 days repeated oral Toxicity studies in rodents
408 90 days repeated oral Toxicity studies in rodents
409 90 days repeated oral Toxicity studies in non rodents
410 90 days repeated Dermal Toxicity
411 90 days inhalation Toxicity study
 412 28/14 days repeated dose inhalation Toxicity study
413 90 days repeated dose inhalation Toxicity study
414 Prenatal Developmental Toxicity study
421 Reproduction /Development toxicity screening test
Important OECD 422 Combined Repeated dose toxicity study with
guideline for Reproduction/Developmental Toxicity screening tes
toxicity 424 Neurotoxicity study in rodents
420 Acute oral Toxicity- fixed dose method
425 Acute oral Toxicity-Up and Down method
451 Carcinogenicity studies
452 Chronic Toxicity studies
453 Combined chronic toxicity/carcinogenic studies
ARRIVE  Animal research: reporting o f In Vivo experiments
Guidelines  Author guidelines of 21 items
IAEC
IAEC –
Composition