Animals are used in various areas of biomedical science such as teaching, research, and testing of drugs. While animal models provide important insights, they have limitations in translating findings to humans due to interspecies differences. To reduce animal use, alternatives such as computer modeling, tissue cultures, and microdosing are being utilized. The 3Rs principle of replacement, reduction, and refinement is also applied to minimize animal pain and distress when animal use is necessary.
Alternative methods to animals testing are the development and implementation of test method that avoid use of live animals or use of less animals in method.
The council directive on protection of animals used for experiments and scientific purpose in article 23
“The commission and member states should encourage
research into development and validation of alternative methods which could provide the same level of information as that obtained in experiment using animals but which involves less animal”.
Alternative methods able to do:
Reduce Refine Replace
collectively called as “The 3Rs Principle”.
Needs for alternative methods
Because in laboratory animals may be:
Poisoned.
Deprived of food water and sleep.
Applied with skin and eye irritants.
Subjected to psychological stress.
Deliberately infected with the infected disease.
Alternative methods to animals testing are the development and implementation of test method that avoid use of live animals or use of less animals in method.
The council directive on protection of animals used for experiments and scientific purpose in article 23
“The commission and member states should encourage
research into development and validation of alternative methods which could provide the same level of information as that obtained in experiment using animals but which involves less animal”.
Alternative methods able to do:
Reduce Refine Replace
collectively called as “The 3Rs Principle”.
Needs for alternative methods
Because in laboratory animals may be:
Poisoned.
Deprived of food water and sleep.
Applied with skin and eye irritants.
Subjected to psychological stress.
Deliberately infected with the infected disease.
This presentation enlists all the studies which are required before submission of IND. It include IND introduction , time period of study ,flowchart showing preclinical studies...
Dermal Irritation and Dermal Toxicity Studies Dinesh Gangoda
Dermal irritation and Corrosion test guidelines 204.
Dermal irritation is the production of reversible damage of the skin following the application of a test chemical for up to 4 hours.
Corrosive reactions are typified by ulcers, bleeding, bloody scabs, and, by the end of observation at 14 days, by discolouration due to blanching of the skin, complete areas of alopecia, and scars. Histopathology should be considered to evaluate questionable lesions. [1]
Dermal corrosion is the production of irreversible damage of the skin; namely, visible necrosis through the epidermis and into the dermis, following the application of a test chemical for up to four hours.[2]
REFERENCES
OECD/OCDE, Test No. 404: ‘‘Acute Dermal Irritation/Corrosion’’, 28 July 2015 OECD Publishing, peris, Page no, 1- 8.
Robert A., Turner., Screening Methods in Pharmacology; 1st edition; Academic press an imprint of Elsevier, pp, 279- 281.
OECD Guideline for testing of chemicals (1981). ‘‘Repeated Dose Dermal Toxicity’’, 21/28- day Study.
REPRODUCTIVE TOXICITY STUDIES, Definition
Introduction, OECD guidelines for reproductive toxicity studies
Principle of the test, Description of Method, Procedure, Experimental Schedule, Data and Reporting, Results, Male Fertility Toxicological Studies
Ms. I. Sai Reddemma.
Department of Pharmacology
Regulatory guidelines for conducting toxicity studies by ichAnimatedWorld
ICH is the “International Conference on Harmonization of
Technical Requirements for Registration of Pharmaceuticals for
Human Use”
ICH is a joint initiative involving both regulators and research based industry representatives of the EU, Japan and the US in
scientific and technical discussions of the testing procedures required
to assess and ensure the safety, quality and efficacy of medicines
Introduction to pre clinical screening of drugsKanthlal SK
Various Techniques and Methods for screening of new chemical entities in preclinical aspects (both invitro & invivo) for effective and safe clinical usage.
Alternative to animal toxicit testing.pptxANANYAPANDEY71
Alternative to animal toxicity studies
pharmacological and toxicological studies
hetcam test
pyrogen test
3R
Refinment,Reduction &Replacement
In-Silico methods
CADD, QSAR
This presentation enlists all the studies which are required before submission of IND. It include IND introduction , time period of study ,flowchart showing preclinical studies...
Dermal Irritation and Dermal Toxicity Studies Dinesh Gangoda
Dermal irritation and Corrosion test guidelines 204.
Dermal irritation is the production of reversible damage of the skin following the application of a test chemical for up to 4 hours.
Corrosive reactions are typified by ulcers, bleeding, bloody scabs, and, by the end of observation at 14 days, by discolouration due to blanching of the skin, complete areas of alopecia, and scars. Histopathology should be considered to evaluate questionable lesions. [1]
Dermal corrosion is the production of irreversible damage of the skin; namely, visible necrosis through the epidermis and into the dermis, following the application of a test chemical for up to four hours.[2]
REFERENCES
OECD/OCDE, Test No. 404: ‘‘Acute Dermal Irritation/Corrosion’’, 28 July 2015 OECD Publishing, peris, Page no, 1- 8.
Robert A., Turner., Screening Methods in Pharmacology; 1st edition; Academic press an imprint of Elsevier, pp, 279- 281.
OECD Guideline for testing of chemicals (1981). ‘‘Repeated Dose Dermal Toxicity’’, 21/28- day Study.
REPRODUCTIVE TOXICITY STUDIES, Definition
Introduction, OECD guidelines for reproductive toxicity studies
Principle of the test, Description of Method, Procedure, Experimental Schedule, Data and Reporting, Results, Male Fertility Toxicological Studies
Ms. I. Sai Reddemma.
Department of Pharmacology
Regulatory guidelines for conducting toxicity studies by ichAnimatedWorld
ICH is the “International Conference on Harmonization of
Technical Requirements for Registration of Pharmaceuticals for
Human Use”
ICH is a joint initiative involving both regulators and research based industry representatives of the EU, Japan and the US in
scientific and technical discussions of the testing procedures required
to assess and ensure the safety, quality and efficacy of medicines
Introduction to pre clinical screening of drugsKanthlal SK
Various Techniques and Methods for screening of new chemical entities in preclinical aspects (both invitro & invivo) for effective and safe clinical usage.
Alternative to animal toxicit testing.pptxANANYAPANDEY71
Alternative to animal toxicity studies
pharmacological and toxicological studies
hetcam test
pyrogen test
3R
Refinment,Reduction &Replacement
In-Silico methods
CADD, QSAR
These presentation includes the information about the replacement of animal experiments (invivo tests) with all the alternative methods like invitro tests and in-silico methods which are used in present century and made the research work easy for pre-clinical and clinical trials.
Alternate animal experiments models for pre and post clinical screening of new drugs.
#Expetrimental_Pharmacology.
#Preclinical Screening methods and testing models.
#Animal_Handeling
Animal Experimentation- Contemporary IssueChandan Saha
Animals have their own rights. They are not puppet of our laboratory. With the help of modern and scientific technology we can change old traditional animal experiment methods.
Today there exists a wide spectrum of views on this subject, ranging from those concerned with animal 'rights' to those who view animals only as a resource to be exploited.
All of thThe five freedoms were originally developed from a UK Government report on livestock husbandry in 1965 (Prof.Roger Brambell) then by Farm Animal Welfare Council (FAWC) In July 1979
Freedom from hunger or thirst by ready access to fresh water and a diet to maintain full health and vigour .
Freedom from discomfort by providing an appropriate environment including shelter and a comfortable resting area .
Freedom from pain, injury or disease by prevention or rapid diagnosis and treatment.
Freedom to express (most) normal behaviour by providing sufficient space, proper facilities and company of the animal's own kind.
Freedom from fear and distress by ensuring conditions and treatment which avoid mental suffering.
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2. Animals are used in science for:
I. Undergraduates teaching to learn physiological
mechanism, anatomy and effect of various drugs
on human body
II. Postgraduate teaching to show effects of various
drugs, to find out the nature of unknown drug
and for bioassay
III. Research to understand the working of body
and processes of disease and health
IV. Research to conduct screening for drugs,
bioassay and for preclinical testing of new drug
Bhupal Noble's University,Udaipur (RAj.)
3. Animal models are used to test possibilities that
would be difficult or impossible to test using the
target species (Humans)
It is mandatory to do extensive toxicological
studies in animals before the candidate drug gets
approval for clinical trials in humans
“There is no doubt that the best test species for
humans are humans. It is not possible to
extrapolate animal data directly to humans due to
interspecies variation in anatomy, physiology and
biochemistry.”
Bhupal Noble's University,Udaipur (RAj.)
6. In the laboratory an animal
maybe:
Poisoned
Deprived of food, water and sleep
Applied with skin and eye
irritants
Subjected to psychological stress
Deliberately infected with disease
Brain damaged, Paralysed,
Surgically mutilated
Irradiated, burned, gassed
Force fed and electrocuted
Bhupal Noble's University,Udaipur (RAj.)
7. It is not possible to replace whole animal models with in vitro systems
to evaluate drug effects on major organ systems.
However, techniques can greatly reduce the number of animals
needed, and refined protocols can improve the design efficiency and
quality of studies,and lessen stress and discomfort experienced by
lab animals.
In order to monitor physiological functions in conscious animals,
survival surgery may be performed to implant catheters, electrodes,
flow probes or other devices.
While chronically instrumented animal models can reduce the
numbers of animals used per study and reduce numbers associated
with acute procedures, these models are resource-intensive to
prepare and maintain.
Generally instrumented animal models can be reused in major organ
systems toxicology (MOST) for studies to evaluate more than one
drug.
Bhupal Noble's University,Udaipur (RAj.)
8. Continued but modified use of animals: 3 R’s
In vitro (test tube) test methods
Tissue cultures techniques:
In-silico (Computer Modelling) techniques:
Computerized patient-drug databases and virtual
drug trials by Computer or mathematical analysis
Computer assisted learning
Non-invasive imaging techniques such as MRIs and
CT Scans
Microdosing
Microorganism based model
Bhupal Noble's University,Udaipur (RAj.)
9. Russel and Burch in 1959 proposed that “if animals were to
be used In experiments, every effort should be made to
replace them with non-sentient alternatives”
They developed the 3R’s strategy which includes:
Refinement- refine experimental methods to decrease
unnecessary pain and trauma to animals
Reduction- reduce the number of animals used in these
experiments
Replacement- replace the animal experiments
Eg.: Computer Simulation Models, In-vitro Methods, Cell
Culture Techniques
Bhupal Noble's University,Udaipur (RAj.)
10. Modified To Reduce Pain & Distress In Animal:
- Providing relief (pain & distress) by giving
drugs like analgesics, anesthetics, tranquilizers,
sedatives.
- By changing procedure
i. Small needle
ii. Use non-invasive techniques like MRI etc.
-Use less sensitive species
-Use smaller dose
- Test can be ended at the earliest feasible time
- Improve housing conditions
Bhupal Noble's University,Udaipur (RAj.)
11. Good planning of studies
i. - Change in experimental design
ii. - Improve methods of data analysis
Sharing research animals
Re-designing Studies
i. - To collect as much information as possible
ii. - Share information
Avoid duplicative testing
i. - By improving communication and co-operation
in the planning & execution of testing
ii. - Sharing data – avoids unintentional repetitions
Bhupal Noble's University,Udaipur (RAj.)
12. Substitution of insentient material in place of
conscious higher animals
Replace higher animals with lower animals.
Replace live animals with dummies for teaching and
dissection purpose.
Could be relative or absolute
Absolute replacement: no need to use animals
Eg,: cell lines, tissue of human or invertebrate cells
and tissues
Relative replacement: humane killing of animals to
provide cells or tissues for in vitro studies
Bhupal Noble's University,Udaipur (RAj.)
13. Instead of using animals, cell and tissue
cultures can be used to test product
ingredients.
Cell culture experiments can show, for
example, the lowest concentration at
which an ingredient causes damage to
cells.
The results enable conclusions to be
drawn about the ingredient’s
compatibility with tissue.
Cell cultures are now also used
routinely to test substances for
mutagenic properties.
A familiar example is the Hen’s Egg
Test, which can be used to test for
mutagenic properties as well.
In vitro biomedical research
entails the maintenance of organs,
tissues (or fragments of organs
and tissues), and cells outside of
the body.
Tissue cultures are additionally
used to test substances for
compatibility with mucous
membranes.
Can be grown as independent cell
lines or preserve the architecture
of the entire organ as organ
culture and tissue culture
Stem cells are also used as invitro
models
Bhupal Noble's University,Udaipur (RAj.)
14. Avian- chick embryos
Rodents- rats and mice ( wild types and
transgenic): embryonic, post-natal and
adult
Human cells:
i. Neural progenitor cells from aborted
foetuses and stem cell lines.
ii. Cord blood derived stem cells.
Bhupal Noble's University,Udaipur (RAj.)
16. In-vitro Pyrogen test
Embryonic stem cell test
Local lymph node assay
for skin sensitization
Clinical skin patch test on
human volunteers
Neutral red uptake assay
Carcinogenicity test
Acute toxicity test
Repeated dose toxicity test
Developmental
neurotoxicity test
Bhupal Noble's University,Udaipur (RAj.)
17. Removal of cells, tissues, or organs from an animal or
plant and their subsequent placement into an
environment conducive to growth.
Types of Tissue Culture:
Bhupal Noble's University,Udaipur (RAj.)
18. PrimaryCultureCellLineCulture Primary cultures are impractical as a source of cells
for high throughput screening b’coz:
i. Difficult to expand the population in culture
ii. Scarcity of human material
iii. High cost
iv. Batch to batch variation
Once the primary culture is sub cultured, it becomes a cell
line
A cell line derived by selection or cloning is referred to as
cell strain.
Cell strain do not have infinite life, as they die after some
divisions.
Types: a) Finite cell line b) Continuous cell line
Bhupal Noble's University,Udaipur (RAj.)
20. Without animal dissection computer generated
simulations are used to predict the various
possible biological and toxic effects of a chemical
or potential drug candidate.
For in vivo experimentation only the most
promising molecules obtained from primary
screening are used.
For example, to know the receptor binding site of
a drug, in vivo experimentation is necessary.
Bhupal Noble's University,Udaipur (RAj.)
21. Computer aided molecular drug design (CADD)
Quantitative structure activity relationships
Computer assisted learning (CAL)
Computer or mathematical analysis
Micro fluidic chips
DNA chips
Organ on chip
Human on chip
Bhupal Noble's University,Udaipur (RAj.)
22. It is used to predict the receptor binding site for a
potential drug molecule.
CADD works to identify the probable binding site
and hence avoids testing of unwanted chemicals
having no biological activity.
We can tailor make a new drug for the specific
binding site and then in final stage animal testing is
done to obtain confirmatory results with the help of
CADD.
Bhupal Noble's University,Udaipur (RAj.)
23. CAL is an interactive computer assisted learning
(CAL) program without involvement of real
experimental tools
CAL had better problem solving attitude &the cost
was much less than the traditional laboratory
practices.
Two softwares are curently used in INDIA:
a.) Ex-pharm b.) X-cology
Bhupal Noble's University,Udaipur (RAj.)
24. Translation of biological effect into a mathematical
equation.
Virtual human organs and virtual metabolism
programmes can now predict drug effects in humans
more accurately then animals can.
Computers design the molecular structure of drugs to
target specific receptors
Eg- Protease inhibitors were designed by computers
and tested in tissue culture and computer models
bypassing animal tests
Bhupal Noble's University,Udaipur (RAj.)
25. Computer programs which can predict the toxicity
of new chemicals or drugs based on their similarity
to more established compounds.
Principle that similar chemicals should have similar
biological properties.
Greater computer power and the ability to generate
large databases have facilitated the development of
these methods and a wide range of models now exist
that cover a variety of toxicities
Bhupal Noble's University,Udaipur (RAj.)
26. Harvard’s Wyss Institute has created "organs-
onchips” that contain human cells grown in a state-
ofthe-art system to mimic the structure and
function of human organs and organ systems
The chips can be used instead of animals in disease
research, drug testing, and toxicity testing and have
been shown to replicate human physiology, diseases,
and drug responses more accurately than crude
animal experiments.
Bhupal Noble's University,Udaipur (RAj.)
28. Chips 2 cm wide and contain a series of tiny chambers
each containing a sample of tissue from different parts
of the body.
The compartments are linked by microchannels
through which a blood substitute flows
The test drug is added to the blood substitute and
circulates around the device
Sensors in the chip feed back information for computer
analysis
This can be used to study the disease process and drug
metabolism
Bhupal Noble's University,Udaipur (RAj.)
29. A ‘microdose’ is defined as less than one hundredth
of the proposed pharmacological dose up to a
maximum of 100 μg
Can be measured in any biological sample including
plasma and urine to determine ADME
Analysed using an accelerator mass spectrometer
(AMS).
Early metabolism data can be obtained before going
into human phase 1 trials.
Allows testing in relevant species.
Bhupal Noble's University,Udaipur (RAj.)
31. Increasing access to the Internet, World Wide Web andkeeping
current with information associated with alternatives to animal
testing is a challenge made easier.
On in vitro and other alternatives to animal testing Internet and
World Wide Web, resources provided guidance and other
information.
The toxicological studies of new cosmetics ingredients should be
present at in vitro. Yet, safety evaluation can be based on in vivo
studies performed before the European ban on the use of animals
came into effect. Before the ban,the evaluations of different
cosmetics ingredients performed by the SCCS are mostly based on
in vivo studies.
At the moment, the total number of in vitro studies is small
compared to that of studies on laboratory animals. The increase in
the use of in vitro methods can be seen in near future.
Though there are some accepted and validated methods, yet there
are no methods for all the studies required, such as repeat dose
toxicokineticsand toxicity others.Bhupal Noble's University,Udaipur (RAj.)