THE ROLE OF ECVAM IN PROMOTING THE REGULATORY ACCEPTANCE OF

ALTERNATIVE METHODS

Dr Andrew Worth

European Centre for the Validation of Alternative Methods (ECVAM)Institute for Health & Consumer Protection

European Commission Joint Research CentreIspra, Italy

Conference on Research into Alternatives to Animal ExperimentationBrussels, 9-10 July 2002

OUTLINE

1) The evolution of regulatory tests- role of ECVAM- role of ESAC

2) The ECVAM validation process

3) Paths toward regulatory acceptance

4) Implications of the chemicals policy for the development and validation of alternative tests

5) The ECVAM report on alternative methods for chemicals testing, and its follow-up

THE ROLES OF ECVAM AND THE ESAC

ECVAM was established by a Communication from the Commission to the Council and the Parliament in October 1991, which laid out the duties of ECVAM

The main duty of ECVAM is to coordinate the validation of alternative tests at the EU level

Validation is the process by which the reliability (reproducibility) and relevance (scientific basis and predictive capacity) of a method are established for a particular purpose

ECVAM has a Scientific Advisory Committee

The most important task of the ECVAM Scientific Advisory Committee (ESAC) to make formal recommendations on the scientific validity of alternative methods, by issuing statements on their applicability

EXAMPLE OF AN ESAC STATEMENT

STATEMENT ON THE APPLICATION OF THE EPIDERMTM

HUMAN SKIN MODEL FOR SKIN CORROSIVITY TESTING

At its 14th meeting, held on 14-15 March 2000 at the European Centre for the Validationof Alternative Methods (ECVAM), Ispra, Italy, the ECVAM Scientific AdvisoryCommittee (ESAC)1 unanimously endorsed the following statement:

Following the review of the results of the ECVAM-funded prevalidation studyon the EpiDermTM skin corrosivity test coordinated by ZEBET, it is concludedthat the EpiDerm human skin model can be used for distinguishing betweencorrosive and non-corrosive chemicals within the context of the EU and draftOECD testing guidelines on skin corrosion.2

The ESAC has been regularly kept informed of the progress of the study, and thisendorsement was based on an assessment of various documents, including, in particular,the report on the results and evaluation of the study prepared for the ESAC,3 and a reporton the study which has been accepted for publication.4

The validation study was conducted in accordance with the general principles laid downin the report of the CAAT5/ERGATT5 workshop held in 1990,6 guidelines contained inthe report of an ECVAM/ERGATT workshop held in 1995,7 criteria laid down byECVAM and the ECB,5,8 criteria recommended at an OECD5 workshop held in 1996,9

and the US ICCVAM report on validation and regulatory acceptance.10

Michael Balls Eva HellstenHead of Unit Head of Unit E.2ECVAM Environment Directorate GeneralInstitute for Health & Consumer Protection European CommissionJoint Research Centre BrusselsEuropean CommissionIspra

21 March 2000

THE EVOLUTION OF REGULATORY TESTS

Stage Key players

1 Research and development Industry, academia

National funding bodies & DG RTD

2 Prevalidation

3 Validation

4 Independent assessment Scientific peer reviewers, ESAC, SCC-NFP

Other expert groups

5 Regulatory acceptance Regulatory bodies e.g. EU Competent Authorities EDQM / European Pharmacopoeia

ECVAM & national validation authorities

OVERVIEW OF THE ECVAM VALIDATION PROCESS

Prevalidation

Formal validation

Catch-up validation

Define structural and performance characteristics and criteria

New test method “Similar” test method

Scientifically validated test method

“Old” test method

Weight-of-evidence review Retrospective validation

Chemicals

1. For nephrotoxicity: Prevalidation of two renal cell lines (LLC-PK1 and MDCK), using trans-epithelial resistance and inulin as permeability endpoints

2. For acute thrombocytopenia: Prevalidation of Colony Forming Unit-Megakaryocyte assay

3. For acute lethal toxicity:Collaborative validation study with NICEATMUse of 3T3 fibroblasts and human keratinocytes to predict acute lethal toxicity in rodents and humans

Biologicals

1. Validation of the Vero cell test for the safety testing of diphtheria vaccines

2. Validation of in vitro methods for pyrogenicity testing (DG RTD contract)

PREVALIDATION AND VALIDATION STUDIES IN PROGRESS

PATHS TOWARD REGULATORY ACCEPTANCE Methods for Chemicals

1) ESAC endorsement of methodOpinions of other Commission scientific committees 2a and 2b

2a) Regulatory acceptance in the EUEU National Coordinators for Test Methods EU Competent Authorities for Directive 67/548/EEC Adoption of Annex V Test Method

2b) Acceptance at OECD level National Coordinators of the OECD Test Guidelines Programme OECD Joint Meeting, EPOC and then OECD Council

Adoption of OECD Test Guideline

REGULATORY ACCEPTANCE - Methods for chemicals

On 4 February 2000, the EU Competent Authorities for Directive 67/548/EEC (on the Classification, Packaging and Labelling of Dangerous Substances) accepted three in vitro methods as replacement methods for the toxicity testing of chemical substances:

1. The rat skin transcutaneous electrical resistance (TER) method for skin corrosion

2. Human skin equivalents (that meet certain criteria) for skin corrosion

3. The 3T3 NRU method for phototoxic potential

These methods have been incorporated into Annex V of Directive 67/548/EEC as Method B.40 (in vitro skin corrosion tests) and Method B.41 (in vitro 3T3 NRU phototoxicity test)

On 29-31 May 2002, the National Coordinators of the OECD Test Guidelines Programme approved Test Guidelines for these methods:

TG 430: in vitro skin corrosion test (TER)TG 431: in vitro skin corrosion test (human skin model)TG 432: in vitro 3T3 NRU phototoxicity test

PATH TOWARD REGULATORY ACCEPTANCE Methods for biologicals

1) ESAC endorsement of method2) Group of Experts of the European Pharmacopoeia: Group 15 (human sera and vaccines) Group 15 V (veterinary sera and vaccines) Group 6 (biological substances) Group 6B (human blood products)3) Publication of draft / revision Monograph proposal4) Commenting period 2 or 5 5) Regulatory acceptance by Council of Europe Countries

European Pharmacopoeia Commission adopts Monograph

Halder (2001). ALTEX 18, Supplement 1, 13-46.

PROGRESS TOWARD REGULATORY ACCEPTANCE Methods for biologicals

1) Production of monoclonal antibodies:

DG ENV has circulated the ESAC statement to the Competent Authorities for Directive 86/609/EEC

2) Batch potency testing of tetanus vaccines for human use

Revised European Pharmacopoeia Monograph, including the ToBI and ELISA tests, has been published in Pharmeuropa for comment

3) Batch potency testing of erysipelas vaccines

Revised European Pharmacopoeia Monograph, including the ELISA test, has been published in Pharmeuropa for comment

THE CHEMICALS POLICY AND ALTERNATIVE METHODS (1)

13 February 2001:

European Commission adopts White Paper on a Future Chemicals Policy• proposes REACH system for new and existing chemicals

• recommends that:“Testing should generally be limited to in vitro methods” for chemicals produced/imported in range of 1-10 tonnes

• Section 3.2 states that:

“… ECVAM’s central role will be maintained, and it is expected that the development of alternative methods will be accelerated. Further research will be carried out both at Community and national level, in order to develop and validate novel testing strategies involving fewer or no animals, while enhancing the relevant information that can be obtained from testing without simultaneously increasing the number of animals involved.”

7 June 2001:

Environment Council concludes (para. 23) that:

“Animal testing should be limited to the level necessary to deliver the objectives of the strategy ….

... In addition to promoting this issue in ECVAM ..., the Community should play a more active role in the OECD, to encourage wider adoption of validated, alternative, non-animal testing methods.”

17 October 2001:

Economic and Social Committee adopts opinion on the White Paper, in which it endorses the commitment to promote non-animal testing

15 November 2001:

European Parliament adopts Schörling report, setting out rules to restrict the number of animal tests and foster the development of alternative methods

THE CHEMICALS POLICY AND ALTERNATIVE METHODS (2)

THE ECVAM REPORT ON ALTERNATIVE METHODS FOR CHEMICALS TESTING

The ECVAM report:

1) was written by ECVAM staff and members of the ECVAM Working Group on Chemicals, with input from the ESAC and other experts

2) describes the current status of alternative methods for human health endpoints

3) makes recommendations for research and development

4) provides a possible time-frame for validation

Worth A.P. & Balls M. (2002). Alternative (non-animal) methods for chemicals testing: current status and future prospects. A report prepared by ECVAM and the ECVAM Working Group on Chemicals. ATLA 30, in press.

Finalised draft available at:http://ihcp.jrc.cec.eu.int/Activities/ACTVali/ACTVali.html

OVERVIEW OF THE ECVAM REPORT

Chapter 1 Background: Chemicals Policy, alternative testing, role of ECVAM

Chapter 2 Principles and procedures of validation

Chapter 3 Scientific basis of chemical risk assessment

Chapter 4 Acute lethal toxicity

Chapter 5 Acute dermal and ocular toxicity

Chapter 6 Skin and respiratory sensitisation

Chapter 7 Biokinetics: barrier function, metabolism, mathematical modelling

Chapter 8 Target organ and system toxicity

Chapter 9 Genotoxicity and carcinogenicity

Chapter 10 Reproductive toxicity

Chapter 11 Endocrine disruption in humans

Chapter 12 Summary of conclusions and recommendations

Possible time-frame for validation

AVAILABILITY OF ALTERNATIVE METHODS (1)

Alternative methods available immediately

1) Skin corrosion (in vitro – replacement)2) Phototoxicity (in vitro – replacement)3) Genotoxicity (in vitro – [partial] replacement) 4) Acute oral toxicity (in vivo – refinement and reduction)5) Skin sensitisation (in vivo – refinement and reduction)6) Developmental toxicity (in vitro – partial replacement)

Short-term prospects for validation (by end 2003)- depending on availability of human and financial resources

1) Acute oral toxicity (in vitro – [partial] replacement)2) Skin corrosion (in silico – partial replacement)3) Eye irritation (in silico – partial replacement) 4) Skin sensitisation (in silico – partial replacement)

AVAILABILITY OF ALTERNATIVE METHODS (2)

Medium-term prospects for validation (by end 2006)- depending on progress and availability of human and financial resources

1) Eye irritation (in vitro – [partial] replacement)2) Skin irritation (in vitro – [partial] replacement; in silico – partial replacement)3) Skin sensitisation (in vitro – [partial] replacement)4) Acute oral toxicity (in silico – partial replacement)5) Genotoxicity (in silico – partial replacement)

Long-term (by end 2010; [partial] replacement )- depending on progress and availability of human and financial resources

1) Respiratory sensitisation 2) Biokinetics3) Target organ / system toxicity, including endocrine disruption4) Chronic toxicity5) Non-genotoxic carcinogencity6) Reproductive toxicity

FOLLOW-UP TO THE ECVAM REPORT: IN SILICO METHODS

A quantitative structure-activity relationship (QSAR) is a mathematical

relationship between a quantitative measure of chemical structure, or a

quantitative measure of a physicochemical property, and a biological activity

ECVAM is currently planning an initiative on the Development, Validation

and Dissemination of QSAR Models, in collaboration with the European

Chemicals Bureau (also part of the IHCP, JRC Ispra)

This will involve:

a) the establishment of a network of experts in the EU Member States, Candidate Countries, the USA, Japan

b) coordination with future OECD activities on QSARs

Special session on QSARs at 34th Joint Meeting, 6 November 2002

FOLLOW-UP TO THE ECVAM REPORT: IN VITRO METHODS

A coordinated action plan is needed within the European Research Area, to ensure the targeted development of new test methods and their subsequent validation

The “development challenge”

ECVAM will contribute by:

1) advising DG Research on priorities for the development of alternative tests relevant to the Chemicals policy

2) establishing a dialogue with industry and academic institutions involved in test development

The “validation challenge”

An enormous task - ECVAM and other validation authorities in the EU Member States will need to share the burden of validation work

ECVAM is best placed to coordinate the overall validation process