1

BTE4231

Animal Models and Disease

States
Chapter 4
 2

Learning outcomes
• Understand the importance of using amial models
• Discuss the various animal models in correlation to drug
discovery and development.
 3

Introduction
• Current in vitro assays – predictive of compounds activity in
vivo
• However, testing in animal models necessary.

WHY?
Regulatory bodies (FDA, etc.) require proof:
i. efficacy of a compound animal models
ii. safety of a compound
 4

Limitations of in vitro assays

1. Incomplete model – information at molecular, cellular or
tissue levels only

2. Unable to replicate compounds ability to modulate

activity of potential drug targets

3. Unable to predict the impact the body has on the

compound
 5

Transition from in vitro assays to animal

models

Transition Effect

Info on what happens to a

Determining the PK compound in the body
profile of a
compound NO info on desired
biological impact

Determine the efficacy of

Determining the PD a compound
of a compound Determine the potency of
a compound
 6

Clinical
Trials
In vivo
assays

In vitro assays
 7

Goal of in vivo efficacy models

• Determine if compound is capable of producing a desired
physiological effect / response
• Compare the impact of a range of doses of the same
compound
• Compare different compounds with each other
 8

Example: Compound to lower

cholesterol
Goal: assess cholesterol levels in an animal model
BUT
•compounds can generate multiple biological responses
•ED50’s for each physiological response can be different

•Pharmacological studies MUST include determining dose

dependency of undesired biological responses
Allows determination of therapeutic window:

positive effect is elicited and negative effect is minimized

•Ratio of these two doses = therapeutic index

 9

Therapeutic Index
• If compound A lowers cholesterol effectively [positive
effect] at 1 mg/kg but causes the heart rate to drop
[negative effect] at 10 mg/kg, then the therapeutic index :
10mg/kg
= 10
1 mg/kg

• Compound A was found to cause hair loss [negative

effect] at 100 mg/kg, making the therapeutic index = 100.

• The lowest therapeutic index is considered.

 10

Sources of animal models

• Limited methods to develop animal models
• Based on several factors:
1. spontaneously occurring mutations and/or
2. selective breeding to develop a population that expresses
OR do not express a trait
3. Using biotechnology – insertion, deletion of genes
4. Using drug effects
5. Behavioral training and responses
6. Surgical and physical alterations to an intact animal
 11

Spontaneously occurring mutations

• E.g. Leptin-deficient mice
• Eat voraciously experiencing
significant weight gain
• Developed conditions similar
to morbid obesity &
type 2 diabetes

• Other examples:
non-obese diabetic mouse
spontaneously hypertensive rat
 12

Using Biotechnology
• SOD1-G93A transgenic mouse model
• Used to study amyotrophic lateral sclerosis (ALS) a.k.a.
Lou Gehrig’s disease
• Incorporation and overexpression of human gene for
superoxide dismutase 1
• Results in mice that display neurodegeneration and
symptoms consistent with ALS
 13

Using Drug effects

• Drug-induced animal models
• E.g. administration of 1-methyl-4-phenyl-1,2,3,6-
tetrahydropyridine (MPTP) in primates and mice
• Leads to rapid destruction of dopamine-synthesizing
neurons in the substansia nigra region of the brain
• Triggers rapid development of
Parkinson’s disease symptoms
– consistent with human conditions
 14

Behavioral training and responses

• Create a situation suitable for assessing the
pharmacological impact of test compounds
• Animal models for depression
• Not possible to determine if a mouse is depressed
• BUT
• Possible to study potential antidepressants in mice by
observing their responses to various situations.
15
 16

Surgical and physical alterations

• Ischemic events can be surgically created by limiting or
blocking blood flow
• To study impact of test compounds on stroke survival
• To study cardiac reperfusion after a heart attack
17
 18

Validity of Animal Models

• Depends on how well the animal model mimics the human
condition
• If there is no correlation no use!
• 3 categories on how well animal models reproduce human
disease:
i. Homologous
ii. Isomorphic
iii. Predictive
 19

Validity of animal models

Category of
Description
animal model

Same causes, symptoms and treatment

options
Homologous Very rare
Construct validity
E.g. non-obese diabetic mice – models for
type 1 diabetes

More common
Same symptoms and treatment
options generally similar
Isomorphic Root cause of disease NOT the same
Face validity – similar but not the
same
E.g. animal models for stroke
 20

Validity of animal models

Category of
Description
animal model

Used when disease condition is poorly

understood
Does not occur in animals
Little or no resemblance to human
Predictive condition
Some facets of the model allow it to be
used as a predictive tool
Predictive validity
E.g. animal models for schizophrenia
 21

Species Selection
• When considering which species to use for an in vivo
study:
• Be aware of limitations of each choice
• In vivo efficacy experiments – key decision points in the
drug discovery and development program
• If the in vivo experiments fail

TERMINATED
 22

Species Selection
• Critical for chosen animal model to give best correlation
with human condition / disease
• Closest animals to humans:

Primates
 23

Species Selection
• Rarely used.
• WHY?
i. Very few available for study
ii. Difficult and expensive to maintain
iii. Large in size – more compound required for testing
iv. More expensive to use
v. Ethical considerations

• ONLY used when no other options available

 24

Species Selection
• More common choices for animal models:

• However, the choice of animal model MUST correlate with

the human disease / condition
• Otherwise, data provided will have little or NO value in
drug discovery and development
 25

Number of animals
• How many is enough?
“Signal strength of a study must exceed the margin of error.
• In theory, ONE animal is sufficient.
The number of
• Practically, animals
there required
are no to “YES”
absolute accomplish thisanswers:
or “NO” will
depend on the strength
errors can occurof the expected signal relative to
the expected margin of error.
• Genetic differences within a population
• Inaccuracies in measuring signal intensities, etc
If the expected signal is strong, the expected margin of
error is low, and fewer animals will be required.
• Therefore, statistical significance is required.
If the signal is weak, the margin of error is large and more
animals will be required for the average in data to be
outside of the margin
EXPLAINof error.THIS!
 26

Animal Models by
Disease Category
 27

Animal Models in Neuroscience

• Depression is clinically important but is difficult to model in
animals
• Described in humans as a pathological complex of
somatic, neuroendocrine, and psychological symptoms
• Replicating these symptoms in animals is not possible
• Development of anti-depressants depend on specific
measurable behaviour predictive of anti-depressant
activity in humans
• Porsolt forced swimming test is used effectively to identify
effective antidepressants
 28

Porsolt forced swimming test

Immobile Swimming
mouse mouse

Compounds with
A model for antidepressant
properties will
depression lengthen time a
mouse will
continue to swim
to attempt to exit
 29

Elevated Plus Maze

• Anxiety – complex disorder in humans
• Measure anti-anxiolytic activity of novel compounds
• Insight into efficacy of compounds such as benzodiazepines
 30

Elevated Plus Maze

 31

Novel Object Recognition Test

• Takes advantage of the natural curiosity of mice
• Can be used to determine the impact of candidate compounds on learning and memory
• Mice are familiarized with an environment
and two objects

• After the acclimation period, one of the known objects is replaced with a new object
• Candidate compounds capable of impacting memory formation (positively or negatively), will influence the amount of time the mouse spends examining the new object.
 32

Novel Object Recognition Test

 33

Animal Models of Cardiovascular

Disease
• Surgical models have been developed to study heart failure (HF) and treatment
of
• Myocardial infarction can be surgically induced by ligating the coronary artery of
animal models
• Dosing with test drugs
is initiated and cardio-
vascular function is
monitored over weeks &
months to determine if
test drug is preventing
heart failure.

•
 34

Cardiac remodeling after HA

 35

Animal Models of Infectious

Disease
• Infectious diseases are a significant health problem
• Antibiotic resistance has increased the intensity of this
issue.
HOW?

• Wide range of infectious agents (bacteria, viruses, fungi,

protozoa) – hundreds of animal models for infections
 36

Animal Models of Infectious

DIsease
• Murine Model of systemic infection
• Important infectious disease model
• Used to identify potential antibacterial agents
• Neutropenia is induced in mice
• Single high dose of bacteria is injected intraperitoneally
• Single dose of candidate compound injected and mouse
is monitored for 48 hours
• Kaplan-Meier survival curves can be used to determine
the efficacy of various doses of of candidate compound in
animal models
 37

Study on Candida species virulence in mouse

models
 38

Limitations of Animal Models in

Infection
• Many infectious diseases are host-limited
• Species-specific interactions important aspect of disease
progression
• might not be replicated in an animal model
• Laboratory strains of pathogens might adapt to laboratory
environment
• Significantly different from clinical isolates
• Changes in gene expression of lab strains could affect results of
preclinical studies
• Transmission pathways
• Very important in disease progression
• Unique pathogenic phenotypes may arise in carrier animals/hosts
 39

Animal Models of Oncology

• Treatment and prevention of cancer is the main concern
of many pharmaceutical companies
• Cancer is multifactorial – which complicates therapy
• Many animal models
 40

Mouse Xenograft Tumor Model

41
42
 43

Conclusion
• Selecting an appropriate animal model is a key aspect of
drug discovery and development
• Drug discovery programs will employ multiple animal
models for a more comprehensive understanding of the
potential clinical drug candidates

• Choosing the wrong animal model can lead to

(i) termination of viable programs due to incorrect data
(ii) progression of drug candidate into clinical trials based
on results that do not correlate well with human
condition
 44

Tutorial 4
1. Why is testing in animal models necessary?
 
2. What is a predictive animal model?
 
3. Why is it necessary to use more than a single animal
model in an in vitro experiment?
 
4. What are some of the limitations of animal models of
infectious disease?