1. International dimension of 3Rs acceptance
…… challenges for industries and regulators
EPAA Betty Hakkert (personal view)

2. Content
REACH
REACH testing strategies
Examples
Challenges

3. REACH Context
1….. This Regulation should ensure a high level of protection of Human Health (HH) and Environment (ENV) as well as….. …… This regulation should also promote the development of alternative methods for the assessment of hazards of substances
16 …..The Regulation is based on the principle that industry should manufacture, import or use substances or place them on the market with such responsibility and care as may be required to ensure that….HH and ENV are not adversely affected.

4. REACH Context: Information Requirements
Annex VI Guidance on fulfilling requirements of Annexes VII-XI
Gather and assessment information
Annex VII to X:
Information Requirements is tonnage dependent (number substances):
≥ 1 tonne/y: Annex VII in vitro (20,000)
≥ 10 tonnes/y: Annex VIII base set (5,000)
≥ 100 tonnes/y: Annex IX additional testing (2500)
≥ 1000 tonnes/y: Annex X additional testing (2700)
Annex XI
General rules for adaptation of the standard testing
Annex XI(1): Testing does not appear scientifically necessary
Annex XI(2): Testing is technically not possible
Annex XI(3): Exposure-driven waiving/testing

5. REACH Context Can be used if:
Annex XI (1) testing does not appear scientifically necessary
Use of existing information, weight of evidence, (Q)SARs, in vitro methods, grouping/read across
Can be used if:
Suitable methods are used
Results are adequate for the purpose of Classification and Labelling and/or Risk Assessment
Adequate and reliable documentation is given

6. REACH Context: Information Requirements
Adequate for Classification and Labelling
Important driver under REACH and several other
legislations (e.g. consumer products, transport)
Adequate for Risk Assessment
Information suited to derive a (semi)quantitative dose descriptor
Adequate for PBT, vPvB assessment

7. REACH Implementation Projects (RIPs)
RIP 3.3 on information requirements provides guidance for industries how to comply with Annexes VI-XI of the Regulation
This guidance was prepared by experts from Industries, Member States and Commission. Joined action!

8. Lessons from RIPs 3.3 (not exhaustive)
Importance of close collaboration between Industries and Member States/Regulators
Realisation there are few, if any, alternative methods that could be used as stand alone replacement for C&L, RA and/or PBT assessment
Further development should focus on regulatory needs and uses
A method stated to be scientifically validated is not necessarily applicable for a given regulatory purpose
There is a need for further development and evaluation of integrated testing strategies and refinement of methods (e.g. EU-project OSIRIS)

9. Lessons RIPs 3.3: example Three in vitro methods for embryotoxicity that have been considered scientifically valid(ated)*
However, these methods are no replacement of OECD 414 and are (as yet) not suited for (certain) regulatory purposes because:
They do (as yet) not provide a dose descriptor suited for RA
They do not include a metabolic system
The evaluations is based on a limited chemical domain
There is a need for further refinement for discrimination between non, weak and strong embryotoxicants
Etc
For use in testing strategies more experience/guidance is needed
By ECVAM advisory committee ESAC

10. Challenges Regulators and Industries
Regulators should be involved throughout the process of method/testing strategy development in order to assure the methods comply with the regulatory needs

11. Anticipated animal use under REACH

12. OECD416
OECD414
OECD421
70% of experimental animals is required for reproductive toxicity testing in REACH
Van der Jagt et al., ECB 2004

13. Retrospective analyses of existing data Can hazard assessment be simplified by changing the testing strategy?
Impact of the second generation in the 2-generation study
(Janer, Hakkert, Slob, Vermiere, Piersma Reprod. Toxicol. 24: (2007))
Comparison of NOELs and critical end points in subchronic versus 2-generation study in the rat
(Janer Hakkert, Piersma, Vermiere, Slob, Reprod. Toxicol. 24: (2007))

14. Toxicological tests and reproductive toxicity
Subacute toxicity test
Subchronic toxicity test
Chronic toxicity test
Reproductive/Developmental toxicity screening tests
Rat two-generation reproductive toxicity test
Rat developmental toxicity test
Rabbit developmental toxicity test
What is the impact on C&L and on NOAEL of the rat two-generation study when a subchronic study is available ?
What is the added value of the 2nd generation

15. Subchronic study: C&L
The subchronic study did not always detect toxicity to fertility  the two-generation studies had an impact on C&L

16. Two-generation vs. Subchronic study: NOAEL
Classified for reproductive toxicity (n = 47)
Not classified for reproductive toxicity (n = 75)
The two generation and the subchronic toxicity tests led to similar overall NOAELs

17. P0 (parental animals) F1 (first generation) F2 (second generation)
What is the impact on C&L and on NOAEL of omitting the second generation in a two-generation reproductive toxicity study ?
Rat two-generation reproductive toxicity test
P0 (parental animals) F1 (first generation) F2 (second generation)

18. Main findings added value 2nd generation
Impact on Classification and Labelling (n=176)
In general no impact on C&L was observed for the second generation
Impact on NOAELs (n=176)
In general no impact on overall NOAEL
Impact leaving out F1 adults (n=176)
Not maintaining F1 until adulthood could lead to “missing” of effects relevant for C&L and for NOAEL
Note: leaving out 2nd generation saves 1200 animals/study

19. CONCLUSIONS present repro-analyses not exhaustive
Data base analysis provides important input in refinement/revision of test guidelines/strategies
Subchronic study is (as yet) insufficient for C&L
2-gen reproduction study may not be needed to define an overall NOAEL
Omitting the second generation in a 2-gen-reproduction study may go without consequences for C&L and NOAEL
Not maintaining F1 until adulthood may miss effects relevant for C&L and NOAEL
The findings support the proposal of an extended 1-generation study (Cooper et al., 2006; Critical Reviews in Toxicology, pg 69-98)
It is important that additional (confidential) data of industry are made available to further substantiate these conclusions and increase the chemical domain

20. Further work repro-analyses
These analyses have been brought into the OECD Test Guideline Programme in the project on the extended F1-generation reproduction study (MS, IND, other stakeholders). Importance of mutual acceptance of the revised method because of the high number of animals involved
Need for more information to increase data base a.o. to obtain clear alerts when to conduct 2nd generation
Work in progress by industries/regulatory bodies on conduct of extended F1-generation study

21. Challenges Regulators and Industries 1
Methods should be developed in order to fulfil the regulatory requirements
Regulators/industries should be involved/consulted in all stages of the development of alternative methods/testing strategies (regulatory needs/capacity building)
Development of high quality data basses is essential for
Refinement in vivo methods (e.g. example repro)
Validation/evaluation alternative methods (e.g. in vitro, “omics”, integrated methods)
Development of models
Application of category approaches/reading across

22. Challenges Regulators and Industries (2)
Challenge to find (acceptable) ways to get/use confidential information from industries to develop/refine methods/testing strategies
In view of the reduction of animals use it is important that methods are widely accepted (e.g mutual acceptance of data of OECD TGs)
Validation/evaluation process should be Transparent and Review Groups should include experts from various field (incl. regulatory knowledge).

23. THANK YOU!