1. Value of in vitro assays in your REACH dossier Frédérique van Acker 18 November 2014

2. Agenda Introduction What are the drivers of in vitro alternatives Regulatory developments Value of in vitro in your REACH dossier New developments Who drives development of these alternatives Status of these new developments Take home message 2

3. Contents of registration dossier For a proper safety assessment data on multiple topics need to be provided Physicochemical properties Ecotoxicology Environmental fate Anticipated human exposure Toxicology  Focus on Toxicological data 3

4. Toxicological building blocks For a full assessment different aspects of toxicology need to be addressed, a.o. Genetic toxicology Acute and repeated dose toxicity Reproduction toxicology Data on metabolism (TK, ADME) Continuous scientific and regulatory developments may lead to considerable changes in the necessary test package for a dossier 4

5. In vivo vs. in vitro Majority of toxicological data are generated in vivo For many endpoints the in vivo data are still considered the golden standard But an increasing number of endpoints can and is addressed in vitro Endpoints genotoxicity, phototoxicity, skin and eye irritation have a tiered approach and performing in vitro assays as first tier is obligatory 5

6. What are the drivers of in vitro alternatives? Public perception Regulatory Image Technological developments in science 6

7. Public perception Increased awareness of CSR and sustainability Animal rights organizations take position Increased societal pressure Higher on political agenda Multiple initiatives emerged to make a good case for alternative methods. EU Horizon 2020 call: PHC-33-2015: New approaches to improve predictive human safety testing (Budget € 88.000.000) 7

8. ECHA’s point of view REACH Annex XI states that an in vivo test must be waived when there is a validated in vitro alternative 2 nd ECHA report on the use of Alternatives, 2014 states: Using animals for testing should be a last resort – only when there are no other scientifically reliable ways of assessing the potential effects on humans or the environment. 8

9. Currently in draft: skin sensitization, cell transformation Regulatory developments in vitro assays 9

10. Cosmetic Products Regulation (EU 1223/2009) In EU complete ban on animal testing for cosmetic products and ingredients since July 2013 Consequences: all endpoints need to be addressed in vitro! Observed ‘workaround’: registration under REACH, pharmaceutical product Article 18 of the Cosmetics Products regulation on animal testing: The toxicological profile of a substance contained in the cosmetic product should be determined for all relevant toxicological endpoints. A particular focus on local toxicity evaluation (skin and eye irritation), skin sensitization, and in the case of UV absorption photo-induced toxicity shall be made. 10

11. Implications Cosmetics regulation Skin penetration → systemic availability → assess systemic toxicity and reprotoxicity How to deal with these complex endpoints? Validated in vitro alternatives not yet available, developments ongoing Regulatory acceptance of alternatives for complex endpoints unclear  Cosmetic industry currently placed in impasse 11

12. Value of these assays in your dossier Multiple in vitro assays are required by ECHA for a REACH dossier Some assays fully replace in vivo assays, others are additional In vitro assays are very useful for mechanistic understanding Our newly developed in vitro assays may even better predict human toxicity, due to use of reconstructed ‘organs’ from human origin. 12

13. New developments: human 3D models 13 www.mattek.com

14. New developments: 3D cell culture → spheroids 14

15. New developments: organ on a chip 15

16. Who drives development of these alternatives? The clients! We see an increase in demand for in vitro, allowing us to continue our innovation for the in vitro market 16

17. Status of these new developments Innovative assays are currently being developed Increased number of in vitro studies regulatory accepted More and more in vitro studies preferred as regulatory replacement 17

18. What is the relevance to us? Regulatory changes determine how dossier is built up Giving sound advise and consultancy is of great value to us We are closely following regulatory developments and ongoing discussions Knowledge of recent changes allows for a tailor-made & cost efficient proposal to compile a dossier 18

19. Take home message Alternative assays are becoming increasingly important in safety assessment Recent advances in technology hold great promise for future assays Organ cultures from human origin may be very useful in mechanistic evaluation and/or prediction for human safety  Keep an eye out for alternative methods ! 19

20. Thank you for your attention! 20 Contact: frederique.vanacker@tno.triskelion.nl