ECVAM s role in making alternative methods available for European legislation

Transcription

1 ECVAM s role in making alternative methods available for European legislation Thomas Hartung & ECVAM Team Institute for Health and Consumer Protection (IHCP) Ispra (Va), Italy

2 ECVAM Animal welfare directive 1986 (86/609/EEC) Established methods validated until 2002 Business plan and reorganisation 2003 responding to 7 th amendment of the cosmetic directive 2003 emerging REACH legislation methods validated staff members Reorganisation 2007 (more drug, vaccine, food, nanotox plus CORRELATE)

3 Cosmetics industry and the 7 th amendment EU: companies, 60 billion turnover EU: new products per year, 25% turnover with products released within last 6 months Marketing ban if testing finished products or not using ECVAMvalidatd methods since 2003 Phasing out ingredient testing with test and marketing bans in 2009 and 2013

4 Chemical industry and REACH EU: companies (96% SME), 590 billion turnover = 33% of world market, 1,7 million employees EU: occupational skin disease cost 3 million work days per year, i.e. about 600 million 86% of toxicological data on old chemicals are lacking REACH will assess chemicals marketed >1 ton/year Expected preregistrations 2009 Expected 70% of testing

5 Reach is coming but how beautiful will it be? Possible finalisation with 2 nd reading on 13 th December 2003: ECVAM business plan: - cut animal numbers by 50% - about 50 validated tests required, i.e. 150 in validation 2005: ECVAM responsible for coordination test strategy development Additional impact of suitable methods

6 Registration Overview Agency start up Pre-registration tonnes CMRs 1+ tonne PBTs/vPvBs (R50-53) 100+ tonnes tonnes tonnes Non-phase-in substances EIF 12 months [6 months] 3 years 6 years 11 years

7 Registration (Q)SARs Waiving: technical exposure Read Across Endpoint Information: Annexes VI-XI TESTING Last resort In-vitro Existing information

8 Validation modular approach Test definition Within-lab. variability Transferability Between-lab.variability Predictive capacity Applicability domain Minimum performance standards Reproducibility Relevance Standardised (INVITTOX protocol) Suitable/Adequate (CORRELATE) Validated (ESAC) Equivalent (CORRELATE)

9 Proposal in the context of the Commission Action plan for Animal Welfare 2006: COmmunity RefeREnce Laboratory for Alternative TEsting sting: : CORRELATEC at ECVAM to speed up validation by instant start to show equivalence of methods similar to validated ones ( me-too ) to define suitable / adequate (not yet validated) methods for REACH

10 Topical toxicity Skin corrosion accepted OECD 2004, 3 rd model validated 2006, 4 th expected 2007 Skin irritation replacement under ESAC review, expected 2007, me-too developments waiting Refinement for eye test under ESAC review, expected alternative for eye test under validation (8 retro- and 2 prospesctive), expected 2008

11 Systemic toxicity ECVAM/NICEATM study to predict starting dose by cytotox under review, human toxicity better predicted than by animal Prediction of non-toxic chemicals (70%) by cytotox under validation, expected 2008 tiered test for dermal and inhalation A-Cute-Tox, expected 2009

12 Mutagenicity / Cancer 2 Cell transformation assays under validation, expected 2008, OECD TG in parallel Micronucleus validated 2006 COMET assay under validation (lead JaCVAM) validation skin models for genotox Workshop false-positives, followup studies with COLIPA CarcinoGenomics omission pos. control, peer-review prepared

13 Sensitisation LLNA cut down approach under validation, expected 2007 Three replacements to start validation 2007 Non-radioactive LLNA under validation Sen-it-i.v.

14 Ecotoxicology Test strategy acute ecotox validated 2006 Fish egg/embryo test prepared for peer-review, expected 2007 Planned validation bioaccumulation with ILSI/HESI

15 Reproductive toxicity Validated embryotoxicity tests to substitute for 2 nd species and priority setting (extended) one-generation study under validation, expected 2008 test battery for alerts under validation 11 endocrine disrupter tests under validation, expected 2008 ReProTect

16 Problem Reproductive Toxicity Dominant test demand: 80% of animal use in REACH (12 million) for substances Lacking capacities and experience (e.g. only 70 two-gen-studies in 25 years) Limited predictve value (60-70% between species) High proportion of false-positive results REACH could lead to many follow-up studies for valuable existing substances Need for new test strategies

17 Calculation consequences of reprotox studies for 1000 chemicals (assuming 5% prevalence, 60% inter-species correlation) 5% 60% Human st spec f- 2 nd spec f- Total f f f f

18 Test strategy development in RIP 3.3 CEFIC management, strong regulator involvement, >200 experts ECVAM coordinated for COM, 28 staff members involved About pages ready Methods under validation already foreseen Avoided (so far) as standard requirements: neurodevelopmental tox., endocrine disruption, respiratory irritation and sensitisation, 2nd species for two-gen study

19 Estimated test animal need for the different endpoints (van der Jagt et al., 2004) Test animal need for different endpoints (% of total test animals needed) -60% Two-generation reprotox Developm. tox study -50% -60% Further mutagenicity x3-66% Avoid: -60% -80% -50% Skin sensitisation Long-term fish tox Developm. tox screening Accumulation Short-term repeated dose Carcinogenicity Sub-chronic tox Long-term repeated tox -100% Short-term fish Possible impact of methods under validation Acute inhalation tox Acute dermal tox -75% Acute oral tox -100% In vivo eye irritation In vivo skin irritation Long-term bird %

20 Re-estimation animal numbers REACH Testing of (1-10 tons), ( tons), 2500 ( tons) and (>1.000 tons) 38 million animals if all tests are carried out Expected 10% existing data and 33% waiving of testing: 23 million animals Impact read-across, chemical classes and (Q)SAR: 18 million animals Impact replacement and reduction methods: 8 million animals

21 Biologicals Vaccines Optimisation of serological models for potency testing of whole-cell pertussis vaccines Humane endpoints: Draft revised monographs on rabies vaccines now includes (ECVAM Workshop 48) Pyrogenicity testing Ongoing ICCVAM pyrogenicity peer review process, Japan? Marine biotoxin testing ECVAM/DG SANCO workshop, joint validation with Comunity Reference Laboratory Marine Biotoxins (CRLMB, Vigo) of functional assay and LC-MS assay for PSP testing Botulinum toxin testing ICCVAM/NICEATM/ECVAM Scientific Workshop; November 2006

22 State of the play 13 research projects with 264 partners (80 million ) 171 methods under validation Expected 100 methods suitable for REACH Currently 125 INVITTOX protocols 9 methods validated in 2006 Expected 40 methods validated until 2009 Impact analysis (peer-reviewed ) suggests 50% animal reduction by alternatives plus 20% by (Q)SAR Draft testing strategies in RIP 3.3 ready to be finalised by

23 Current Activities & Prospects Online availability of the whole DB-ALM data contents as an Oracle based database service (29th October 2006) Data collections on Reproductive Toxicity and Eye Irritation (>2 years) Extension of DB-ALM to include information on computational toxicology including QSARs ECVAM website, INVITTOX protocols, EBT platform

24 Complementary Activities ECVAM website visitors, visits, 670 newsletter registrations (Nov 2006) Details on in-house and external collaborative activities Validated Methods Main Publications (Workshop & Task Force Reports) Registration to the ECVAM Newsletter Thesaurus (open source list) Results of an international ECVAM Task Force on Alternatives Databases Pilot project to identify terminology most commonly used by scientists active in the animal alternatives area for indexing purposes

25 Information Content In vitro methods Method Summary Descriptions INVITTOX Protocols Evaluation Studies Formal validation Studies Test Results Bibliographic References Computational modeling - (Q)SARs Model Summary Descriptions Validation Exercises Training- and Test sets of chemicals

26 Impact Analysis 4667 registered users from 67 countries 7% 20% 44% 29% (until October 2006) Europe Americas Asia Others

27 User Profiles 10% 14% 31% 45% Government Academia Industry Others

28 Highlight 2006

29 Europe goes alternative 2nd Conference, Brussels, 18th of December 2006 Hosted again by Commissioners G. Verheugen (DG ENTR) and J. Potocnik (DG JRC / DG RTD) European Partnership (7 trade associations, 22+ companies) Action programme published Mirror group established Various workshops

30 The validation dilemma What we do not know Classical Validation What we would like to know

31 Opinion leader meeting on stakeholder perception of alternatives Validation of alternative tests is one of the rare examples of quality assurance in biomedical research (relevance, not only reproducibility) New concept: Evidence-based medicine goes in vitro! Evidence-based Toxicology Tools: - Validation studies - Quality assurance (GLP, GCCP) - Systematic review & Meta-analysis

32 Clinical medicine is moving from an experienceand expert-driven to a scientific discipline Learning from experience may be nothing more than learning to make the same mistakes with increasing confidence. Petr Skrabanek, James McCormick Follies and Fallacies in Medicine Tarragon Press, Glasgow, 1989

33 Archie Cochrane British Physician and Epidemiologist ( ) 1979: It is surely a great criticism of our profession that we have not organized a critical summary, by specialty or subspecialty, adapted periodically, of all relevant randomized controlled trials.

34 AIM Systematic Reviews - prospective planning - thorourgh literarture search - evaluation of quality - reproducible analysis - identical formats The Cochrane Database of Systematic Reviews

35 Diagnosis: toxic! - Trying to apply approaches of clinical diagnostics and prevalence in toxicology considerations (Tox Sci 85, , 2005) Thought starter: - Healthy European without HIV risk factors: Prevalence of infection is 1: The result of 99.9% accurate test is positive - Testing people with this test will result in 1 real-positive but 10 false-positive - Probability of HIV infection: 1/11 = 9% In toxicology prevalences and accuracy are often unknown

36 Diagnosis: toxic! - The impact of prevalence Prevalence : The proportion of individuals substances in a population havingtoxic being a disease Predictive capacity: specificity and sensitivity vs. NPV and PPV

37 EYE IRRITATION in vivo Haseman 2005: 5 15% underprediction of GHS-1 (= R41) as inter-animal variance within a single test < within-laboratory variance (day-to-day variances, performers) < between-laboratory variance (laboratory set-up, animal strains, performance) < predictive capacity for human health effect No comprehensive analysis of high-quality data exists; Review in preparation our tools are not perfect

38 The prevalence concept Prevalence of eye irritants in the New Chemicals Database (n = 2497) R41 R36 Negative predictive value R41: 85% accuracy: 97% Positive predictive value R41: 85% accuracy: 51% Every test (in vivo or in vitro) will have many false-positives and few false-negatives Identify the negatives

39 Diagnosis: toxic! - Conclusions Toxicological tests are diagnostical tools parallels to clinical/diagnostic medicine making use of evidence-based methodology Account for reference standard imperfectness Systematic prevalence assessment estimation of a test s diagnostic value e.g. in low prevalence situation or in the context of testing strategies

40 ECVAM structure FP7 policy horizontal database cosmetics (agro-) chemicals admin. support EBT external collaboration drugs consumer products infrastructure

41 Toward an evidence-based toxicology Toxicology requires a critical review of its toolbox Validation is first of all a mean of quality control, which needs to be combined with quality assurance (GLP in vitro, GCCP) and structured reviews/meta-analysis Acceptance of novel approaches requires to understand the limitations of the current ones Article: S. Hoffmann & T. Hartung Toward an evidencebased toxicology Human and Exp. Tox. 2006, 25: Next steps: Current development of an internet platform International Forum Oct 2007, Como, Italy For further information: ebt.forum@jrc.it

42 Globalisation of 3R

43