ALTERNATIVE ANIMAL TOXICITY STUDY .pptx

Alternatives animal testing are development and implementation of test methods that avoid the use of live animals.
Human biochemistry, physiology, pharmacology, and endocrinology and toxicology has been derived from animal models.10-100 millions of animals are using for experimentation in a year.
Animals used experimentation distributed among zebra- fish to primates.
Vast majority of animals are sacrificed at end of research programme.The use of animals can be further subdivided according to the degree of suffering
Minor animal suffering:- observing animals in behavioral studies, single blood sampling, Immunization without adjutants, etc.
Moderate animal suffering:- repeated blood sampling, recovery from general anesthesia, etc.

1. SUBJECT : TOXICOLOGICAL SCREENING METHODS (II) TOPIC : ALTARNATIVE METHODS TO ANIMAL TOXICITY TESTING SAMIR PANDA ( I I ND SEM, M.PHARM ) DEPARTMENT OF PHARMACOLOGY

2. INTRODUCTION Alternatives animal testing are development and implementation of test methods that avoid the use of live animals. Human biochemistry, physiology, pharmacology, and endocrinology and toxicology has been derived from animal models.10-100 millions of animals are using for experimentation in a year. Animals used experimentation distributed among zebra- fish to primates. Vast majority of animals are sacrificed at end of research programme.The use of animals can be further subdivided according to the degree of suffering Minor animal suffering:- observing animals in behavioral studies, single blood sampling, Immunization without adjutants, etc. Moderate animal suffering:- repeated blood sampling, recovery from general anesthesia, etc. Severe animal suffering:- LD50% test, starvation vaccine potency tests, etc. 9/12/2023 2

3. HISTORY In 1824 the organization for animal rights: royal society for the prevention of cruelty to animals In 1876, an act for prevention of cruelty to animal was formed in the UK In India prevention of cruelty to animals (PCA) Act came in 1960 and amended in 1982 In the late 1990s, the committee for the purpose of control and supervision of experiments on animals (CPCSEA) came into existence. 3

4. Regulatory agencies 1989:- ZEBET in Germany. 1993:- ECVAM (European). 1997:- ICCVAM (US) consists of 15 research and regulatory agencies among which the Environmental Protection Agency 1999(EPA), the Food and Drug Administration 2001 (FDA), and the Agency for Toxic Substances and Disease Registry 2003 (ATSDRO). 2005:- JaCVAM (Japanese). 2011:- BraCVAM (Brazil) 9/12/2023 SAMIR PANDA 4

5. LEGISLATION EU directive 2010/63/EU:- on January 1,2013, EU directive 2010/63/EU member states EU cosmetic regulation:- It establishes prohibitions against. testing finished cosmetic products and cosmetic ingredients on animals (testing ban), (b) marketing in EU finished cosmetic products. EU chemical policy: REACH the Registration, Evaluation, Authorization and Restriction of Chemicals came In force in 2007. EU test methods regulation: development and validation of alternative techniques. 9/12/2023 SAMIR PANDA 5

6. REASONS FOR DEVELOPMENT OF ALTERNATIVE ANIMAL TESTING Economic and efficiency play a key role. The "In vitro" testing provide toxicity information in a cost effective and time saving manner. These can also increase the efficiency of whole study and decrease the number of animals required for toxicity. Invitro testing human cell lines have been useful in securing relevant information for human risk assessment thereby opening up opportunities to explore responses to existing and emerging therapies human cell lines can be used for the tumor and some other chronic disease. 9/12/2023 SAMIR PANDA 6

7. INTERNATIONAL ORGANIZATION ICATM International cooperation on alternative test methods(ICATM): on April 27,2009 United states, Canada, japan and EU. Reduction of the number of animals required for consumer product safety testing worldwide. OECD Organization for Economic Co-operation and Development. More than 35 member countries worldwide. Test with OECD test guidelines and principles of Good Laboratory Practice (GLP), shall be accepted. 9/12/2023 SAMIR PANDA 7

8. ACCORDING TO OECD Complete replacement of the animal experiment TG 428 skin absorption: in vitro method TG 430 In vitro skin corrosion:-transcutaneous electrical resistance(TER) TG 431 In vitro skin corrosion:- human skin model test TG 432 In vitro 3T3 NRU phototoxicity test TG 437 Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants. It is evaluated by ICCVAM, ECVAM, JACVAM. 9/12/2023 SAMIR PANDA 8

9. TG 439 Invitro skin irritation: reconstructed human epidermis ( RhE ) test method acute toxic class method Reduction in the number of animals and stress of the laboratory animals G 420 acute oral toxicity --fixed dose procedure 1423 Acute oral toxicity acute toxic class method TG425 Acute oral toxicity up and down procedure TG 429 skin sensitization local lymph node assay TG 486 Acute inhalation toxicity 9/12/2023 9

10. ALTERNATIVE TESTS In silico computer simulation methods In vitro cell culture techniques Using fewer animals Microdosing Epidemiologic and clinical studies and genetic monitoring 9/12/2023 SAMIR PANDA 10

11. IN SILICO COMPUTER SIMULATION METHODS There are many software available in the public domain. Computers can predict the toxicity of chemicals, including: Their potential to cause cancer or birth defects Based on their molecular structure Lhasa limited:- promotes the sharing of data and knowledge. It has developed software tools such as:- DEREK (Deductive estimation of risk based on existing knowledge) Meteor:- knowledge based expert system that predict toxicity and metabolism of chemical 9/12/2023 SAMIR PANDA 11

12. Vitic :- its chemical-based intelligent toxicity database These software tools include illustrations on:- The use of data entry and editing tools for the sharing of data and knowledge within the organization II. Use of proprietary data to develop non-confidential knowledge that can be shared between organization III. The use of shared expert knowledge to refine predictions IV. The sharing of proprietary data between organizations through the formation of data sharing. The use of proprietary data to validate predictions Recent studies show that the use of the "in silico" method is the way forward for assessing the genotoxicity and potential carcinogenicity/repeat dose general toxicology of large numbers of chemicals. There are a number of other in silico methods which have been developed and validated, for example:- Leadscope , mcase , topkat , Toxfree, Osiris, some QSAR models this software was developed by the FDA with proprietary databases 9/12/2023 12

13. INSILCO METHODS IN DRUG DISCOVERY Molecular docking, QSAR, pharmacophore mapping – Molecular docking Docking is the computational determination of binding affinity between molecules(protein structure and ligand). 9/12/2023 SAMIR PANDA 13

14. COMFA:- Comparative molecular field analysis The biological activity of a molecule is dependent on the surrounding molecular fields(steric and electrostatic fields) COMSIA:- Comparative molecular similarity index analysis Include more additional field properties:- steric, electrostatic, hydrophobic, hydrogen bond donor, hydrogen bond acceptor Can offer a more accurate SAR than COMFA 9/12/2023 SAMIR PANDA 14

15. IN VITRO CELL CULTURE TECHNIQUES In vitro cell culture is currently the most successful, and promising, alternative to animal use. Isolated cells, tissues and organs can be prepared and maintained in culture by methods that preserve properties and characteristics of the same cells, tissues and organs in vivo. Cell culture was 1st successfully undertaken by Ross Harrison in 1907. Cells and tissue cultures are used to screen new therapies and to test for product safety Different types of cell grown in culture includes connective tissue elements such as fibroblasts, skeletal tissue, cardiac, epithelial tissue(liver, breast, skin, kidney) and many different types of tumour ells. Genetic microarrays are being used to predict liver toxicity by measuring gene expression in human liver cells. 9/12/2023 SAMIR PANDA 15

16. Cultured cells have also been developed to create monoclonal antibodies thereby replacing use of animals required to undergo a procedure likely to cause pain and distress 9/12/2023 16 Advantages of in vitro studies:- Small quality of test substance is needed. Experiment is under controlled conditions Result is obtained quickly. No major infrastructure is required. Disadvantages of in vitro studies:- In vitro dose-response not available No systemic effect is studied Organ specificity lacking Chronic and long-term effects could not be studied Transportation of material not easy QUESTIONS Discuss the alternative methods to animal toxicity testing ?

17. REFERENCE Araújo GL de, Campos MAA, Valente MAS, Silva SCT, França FD, Chaves MM, et al. Alternative methods in toxicity testing: the current approach. Braz J Pharm Sci [Internet]. 2014;50(1):55–62. Alternative methods to animal toxicity testing [Internet]. Slideshare.net. [cited 2023 Sep 14]. Available from: https://www.slideshare.net/SanchitDhankhar/alternative-methods-to-animal-toxicity-testing-249329802 Goldberg AM, Frazier JM. Alternatives to animals in toxicity testing. Sci Am [Internet]. 1989;261(2):24–31. Vinardell M, Mitjans M. Alternative methods to animal testing for the safety evaluation of cosmetic ingredients: An overview. Cosmetics [Internet]. 2017 [cited 2023 Sep 14];4(3):30. 9/12/2023 SAMIR PANDA 17

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