Alternate animal experiments ( Basic Introduction )

Alternate animal experiments models for pre and post clinical screening of new drugs.
#Expetrimental_Pharmacology.
#Preclinical Screening methods and testing models.
#Animal_Handeling

1. ALTERNATE ANIMAL ALTERNATE ANIMAL
EXPERIMENTSEXPERIMENTS
Presented by :- Presented by :-
Deepak Chandra JoshiDeepak Chandra Joshi
M.Pharma (Pharmacology)M.Pharma (Pharmacology)
Department of Pharmaceutical Sciences Department of Pharmaceutical Sciences
Bhimtal (Nainital)Bhimtal (Nainital)

2. Table of Content :-Table of Content :-

IntroductionIntroduction

Need for alternative to animalNeed for alternative to animal

Laws and regulationsLaws and regulations

Alternative experiments methodsAlternative experiments methods
1.1.RefinementRefinement
2.2.Reduction Reduction
3.3.ReplacementReplacement

In-Vitro MethodsIn-Vitro Methods

3. IntroductionIntroduction

Alternative animal experiment are the develepmAlternative animal experiment are the develepm
-ent and Implementation of test method’s that -ent and Implementation of test method’s that
avoid the use of life animals. avoid the use of life animals.

The primary goal of alternative animal experimeThe primary goal of alternative animal experime
-nt is the minimize pain which is suffer by -nt is the minimize pain which is suffer by
animal during the experiments.animal during the experiments.

4. Animal are used in Science for:-Animal are used in Science for:-

UG teaching to learn physiological mechanism UG teaching to learn physiological mechanism
,anatomy and effect of various drugs on human ,anatomy and effect of various drugs on human
body.body.

PG teaching to show effects of various drugs to PG teaching to show effects of various drugs to
find out the nature of unknown drug and it’s find out the nature of unknown drug and it’s
effect or for bioassayeffect or for bioassay

In research to understand the working of body at In research to understand the working of body at
disease and healthy statedisease and healthy state

In research to conduct screening for drugs In research to conduct screening for drugs
bioassay and pre clinical testing of new drugs bioassay and pre clinical testing of new drugs

5. 
Animals are used to test possibilities that would be Animals are used to test possibilities that would be
difficult or impossible to test using the target species difficult or impossible to test using the target species
(humans).(humans).

It is mandatory to do extensive toxicological studies in It is mandatory to do extensive toxicological studies in
animals before the drug gets approval for clinical trials animals before the drug gets approval for clinical trials
in humans.in humans.

““There is no doubt that the best test species for humans There is no doubt that the best test species for humans
are humans. It's not possible to extrapolated animal data are humans. It's not possible to extrapolated animal data
directly to humans you to inter species variation in directly to humans you to inter species variation in
anatomy ,physiology and biochemistry”.anatomy ,physiology and biochemistry”.

6. Needs for Alternatives :-Needs for Alternatives :-

In Laboratory an animal may be….In Laboratory an animal may be….
1. Poisoned.1. Poisoned.
2. Deprived of food ,water and sleep.2. Deprived of food ,water and sleep.
3. Applied with skin and eye irritants.3. Applied with skin and eye irritants.
4. Subjected to physiological stress.4. Subjected to physiological stress.
5. Deliberately infected with disease.5. Deliberately infected with disease.
6. Brain damaged, paralysis, surgical mutilated.6. Brain damaged, paralysis, surgical mutilated.

7. 
Disadvantages of animal experiments :-Disadvantages of animal experiments :-
1.1.Pain, distress and unethical behavior to Pain, distress and unethical behavior to
animals.animals.
2.2.Requirement of skilled manpower.Requirement of skilled manpower.
3.3.Time consuming protocol.Time consuming protocol.
4.4.High Cost.High Cost.

8. Laws and Regulations:-Laws and Regulations:-
YEARYEAR LAWLAW
19601960Prevention of cruelty to animal act. (amend.1982).Prevention of cruelty to animal act. (amend.1982).
19641964Committee for the purpose of control and Committee for the purpose of control and
supervision of experimental animal (CPCSEA).supervision of experimental animal (CPCSEA).
19921992Indian national science academy. “Guidelines for Indian national science academy. “Guidelines for
care and use of animals in scientific research.care and use of animals in scientific research.
20012001Indian Council of medical Research (ICMR).Indian Council of medical Research (ICMR).
““Guidelines for use of laboratory animals in Guidelines for use of laboratory animals in
medical collages”.medical collages”.
20092009MCI amendment to use alternatives to replace MCI amendment to use alternatives to replace
animal experiments.animal experiments.
20132013UGC guidelines for discontinue of animal UGC guidelines for discontinue of animal
experiments in zoology or life science in a phased experiments in zoology or life science in a phased
manner.manner.

9. Modified use of animals:-Modified use of animals:-

Russel and Burch in 1959 proposed that ….Russel and Burch in 1959 proposed that ….
“ “If animals were to be used in experiments every effort If animals were to be used in experiments every effort
should be made to replace them with non-sentient should be made to replace them with non-sentient
alternatives”.alternatives”.

They developed the 3R strategy which includes:-They developed the 3R strategy which includes:-
1.1.Refinement:-Refinement:- refine experimental methods to decrease refine experimental methods to decrease
unnecessary pain and trauma to animals.unnecessary pain and trauma to animals.
2.2.Reduction:-Reduction:- reduce the no. of animals used in these reduce the no. of animals used in these
experiments.experiments.
3.3.Replacement:-Replacement:- replace the animal experiments .replace the animal experiments .
ex.Computar stimulation model , in-vitro method , cell ex.Computar stimulation model , in-vitro method , cell
culture technique.culture technique.

10. 1.Refinement1.Refinement

Setting the earliest possible end point.Setting the earliest possible end point.

Using appropriate analgesics and anesthetics for Using appropriate analgesics and anesthetics for
painful procedure.painful procedure.

Use proper handling technique for animals.Use proper handling technique for animals.

Adequate training prior to performing Adequate training prior to performing
experiment.experiment.

Ensure drug dose are correct and srug are not Ensure drug dose are correct and srug are not
expired.expired.

Performed surgeries and procedure aseptically to Performed surgeries and procedure aseptically to
prevent infection.prevent infection.

11. 2.2.ReductionReduction

Perform pilot studies.Perform pilot studies.

Design studies to use animals as their own Design studies to use animals as their own
control. ex. Cross over study.control. ex. Cross over study.

Gather data for more than one experiment Gather data for more than one experiment
concurrently.concurrently.

Consult with statistician and use minimum no. of Consult with statistician and use minimum no. of
animals.animals.

Minimize variables such as disease , diet , stress, Minimize variables such as disease , diet , stress,
genetics.genetics.

Use appropriate species of animals. Use appropriate species of animals.

12. 3.Replacement 3.Replacement

Substitution of insentient material in place of Substitution of insentient material in place of
conscious higher animals.conscious higher animals.

Could be relative or absolute. Could be relative or absolute.

Replace higher animals with lower animals .Replace higher animals with lower animals .

Replace live animals with dummies for teaching Replace live animals with dummies for teaching
and dissection purpose .and dissection purpose .

Use computer simulation and in-vitro methods .Use computer simulation and in-vitro methods .

Use cell culture and tissue culture.Use cell culture and tissue culture.

13. 1. In-Vitro Models:-1. In-Vitro Models:-

In-Vitro biomedical research entails the In-Vitro biomedical research entails the
maintenance of organs, tissues (fragments of maintenance of organs, tissues (fragments of
organs and tissues), and cells outside the body.organs and tissues), and cells outside the body.

Can be grown as independent cell lines or Can be grown as independent cell lines or
preserve the architecture of the entire organ as preserve the architecture of the entire organ as
organ culture and tissue culture.organ culture and tissue culture.

Stem cells are also used in in-vitro models.Stem cells are also used in in-vitro models.

14. Source of tissue for in-vitro methods:-Source of tissue for in-vitro methods:-

Avian – Chick embryosAvian – Chick embryos

Rodents – rats and mice (wild types and adult traRodents – rats and mice (wild types and adult tra
-nsgenic). Embryonic, post-natal and -nsgenic). Embryonic, post-natal and
and adult.and adult.

Human – 1. Neural progenitor cells from aborted Human – 1. Neural progenitor cells from aborted
fetuses and stem cell line.fetuses and stem cell line.
2. cord blood derived stem cells.2. cord blood derived stem cells.

15. Types of in-vitro System:-Types of in-vitro System:-
Cell Culture
Cell Lines
Primary
culture
Organ
architecture
preserved

16. In-vitro methods :-In-vitro methods :-

In-vitro pyrogen test .In-vitro pyrogen test .

Embryonic stem cell test .Embryonic stem cell test .

Local lymph node assay for skin sensitization.Local lymph node assay for skin sensitization.

Clinical skin patch test on human volunteers.Clinical skin patch test on human volunteers.

Neutral red uptake assay.Neutral red uptake assay.

Carcinogenicity test.Carcinogenicity test.

Acute toxicity test.Acute toxicity test.

Repeated dose toxicity test.Repeated dose toxicity test.

Development neurotoxicity test.Development neurotoxicity test.

17. 2.Microorganism based model:-2.Microorganism based model:-

Tetrahymena pyriformis – a ciliate protozoan Tetrahymena pyriformis – a ciliate protozoan
being used to study the effects of anesthetics on being used to study the effects of anesthetics on
metabolism.metabolism.

Salmonella typhimurium – bacteria used in Salmonella typhimurium – bacteria used in
mechanistic studies in genetics as well as the mechanistic studies in genetics as well as the
Ames mutagenicity test.Ames mutagenicity test.

18. 3.3.In-chemico testing :-In-chemico testing :-

The toxic potential of substances can sometimes be The toxic potential of substances can sometimes be
detected using relatively simple chemistry based detected using relatively simple chemistry based
methods and not requiring human cells.methods and not requiring human cells.
ex. HPLCex. HPLC

Direct peptide relativity assay – used to assess whether a Direct peptide relativity assay – used to assess whether a
chemical or cosmetics will cause allergy.chemical or cosmetics will cause allergy.

The test works by mimicking a key step in the The test works by mimicking a key step in the
development of allergies – the binding of proteins development of allergies – the binding of proteins
found in the skin to substance.found in the skin to substance.

If proteins bind to the substance them its very unlikely If proteins bind to the substance them its very unlikely
that it will cause an allergic reaction.that it will cause an allergic reaction.

19. 4.4.In-silico Models :-In-silico Models :-

Computer aided molecular drug design.Computer aided molecular drug design.

Quantitative structure activity relationships.Quantitative structure activity relationships.

Computer assisted learning.Computer assisted learning.

Computer or mathematical analysis.Computer or mathematical analysis.

Microfludic chipsMicrofludic chips

DNA chips.DNA chips.

Organ on chipOrgan on chip

Human on chipHuman on chip

20. 5.5.Computer assisted learning (CAL) :-Computer assisted learning (CAL) :-

CAL deals with a range of software packages CAL deals with a range of software packages
which simulate the animal experiments.which simulate the animal experiments.

Two software's are currently used in india.Two software's are currently used in india.
1.1.Ephram – developed by JIPMER, India.Ephram – developed by JIPMER, India.
2.2.X-cologyX-cology

21. ExpharmExpharm

Contains programs on……..Contains programs on……..
1.1.Effects of drugs on rabbit eye.Effects of drugs on rabbit eye.
2.2.Bio assay of histamine using guinea pig ileum.Bio assay of histamine using guinea pig ileum.
3.3.Effects of drug on frog heart.Effects of drug on frog heart.
4.4.Effects of drugs on dog blood pressure and HREffects of drugs on dog blood pressure and HR
5.5.Effects of drugs on cillary movement of frog Effects of drugs on cillary movement of frog
esophagus.esophagus.

The user can conduct experiment and collect data.The user can conduct experiment and collect data.

Each program can be run in two modes :-Each program can be run in two modes :-
1.1.Tutorial modeTutorial mode
2.2.Examination modeExamination mode

22. THE ENDTHE END
DEEPAK JOSHIDEEPAK JOSHI
M.PHARMA M.PHARMA
(PHARMACOLOGY)(PHARMACOLOGY)