Dansk repræsentant i ESAC, følgegruppe til EU s Forskningscenter vedr. alternativer til dyreforsøg.
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- Augusta Bundgaard
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1 Alternativer til dyreforsøg - de tre R er Med hovedvægt på - REPLACEMENT Min baggrund er cand.scient. i biokemi og PhD i biomedicin. Eksperimentel erfaring med dyreforsøg og in vitro metoder med anvendelse af celler fra mennesker og dyr. Dansk repræsentant i ESAC, følgegruppe til EU s Forskningscenter vedr. alternativer til dyreforsøg. Koordinator af EU-projekt, som bl.a. etablerer moderkage system til undersøgelse af transport af stoffer fra moder til foster. Miljømedicin Københavns Universitet
2 Dyreforsøg giver os viden om stoffers virkning i hele organismer til forskel fra isolerede celler eller organer om Akut toksicitet Irritation Langtidseffekter - kræft, mutagen, reproduktion
3 Dyreforsøg anvendes i dag ved Testning af nye kemikalier på markedet Testning af eksisterende kemikalier på markedet Udvikling af lægemidler m.m. Testning af lægemidler Testning af andre forbrugerprodukter Områder hvor vores viden er utilstrækkelig - feks forskelle i følsomhed mellem børn og voksne
4 What is an alternative method? Three types of alternative methods: Reduction alternatives obtain a comparable level of information from the use of fewer animals, or more information from the same number of animals Refinement alternatives minimise pain, suffering and distress Replacement alternatives permit a given purpose to be achieved without using animals Reference: Russell, W.M.S. & Burch, R.L. (1959). The Principles of Humane Experimental technique. Methuen, London.
5 Hvilke alternativer findes inden for replacement Struktur analogi overvejelser Analogi med beslægtede stoffer Teste på menneske (eller dyre) celler Teste på forsøgspersoner, evt patienter
6 Væsentlige strukturelle enheder, der anses for mistænkelige strukturer i forhold til genotoksisk effekt Ashby, 1991
7 Eksempel på replacement: Erstatning af brug af kaniner med cellekulturer til testning af vacciner for tilstedeværelse af urenheder Kaninen (og mennesker) vil reagere med temperatur stigninger, hvis der er urenheder i form af pyrogener tilstede Cellekulturen vil reagere med udskillelse af målbare komponenter, hvis der er urenheder til stede
8 Eksempel på replacement: RANTIV: Risk Assessment for Neurotoxins Tested In Vitro
9 Hvorfor går det så langsomt med udviklingen af alternativer til dyreforsøg Validering tager tid Der er ikke stor anerkendelse af alternative i den videnskabelige verden Der mangler incitamenter i form af styring fra bevillingsgivere
10 ECVAM AND PROTECTION OF HEALTH OF THE CITIZEN Research Development Other interested parties, health groups, consumers, animal welfare groups, etc... Implementation Regulatory Acceptance Prevalidation Independent Review DG Research Validation DG Environment DG Enterprise EU Member States Industry ECVAM ECVAM DG Health and Consumer Protection
11 What does it mean to validate a method? to establish the reliability and relevance of the method for a particular purpose Reliability: reproducibility of results within and between laboratories and over time Relevance: scientific value and practical usefulness Purpose: the intended application of the procedure This definition applies to both alternative AND animal methods References: (1) The Amden CAAT/ERGATT workshop (ATLA 18, , 1990) (2) Frazier s report to the OECD (OECD Environment Monograph no. 36)
12 What is an alternative (replacement) test? An alternative test can be regarded as the combination of a test system and a prediction model. Reference: Archer et al. (1997). ATLA 25, ALTERNATIVE TEST TEST METHOD = + PREDICTION MODEL A prediction model (PM) is an explicit decision-making rule for converting the results of one or more alternative tests into a prediction of an in vivo endpoint.
13 Stages in the evolution of regulatory tests 1 Research and development Supporting role of ECVAM 2 Prevalidation 3 Validation Leading role of ECVAM (EU Competent Authorities) 4 Independent assessment Scientific peer review, ESAC 5 Regulatory acceptance Responsibility of other bodies Timescale: Prevalidation acceptance typically 6 years
14 Criteria for test development For entry into (pre)validation, a test should be accompanied by: A definition of its scientific purpose and proposed practical application A description of its scientific basis The case for its relevance, including its advantages compared with other tests An optimised protocol, including: - standard operation procedures - a specification of endpoints, endpoint measurement, derivation and expression of results, and their interpretation, via a prediction model - the inclusion of adequate controls A statement about limitations/domain of applicability Evidence of reproducibility (within labs, and if available, between labs) Reference: Balls & Fentem (1999). Toxicology in Vitro 13,
15 Post-validation Independent assessment Publication of the study in a peer-reviewed journal Assessment of the outcome by an independent expert panel Statement on the validity Regulatory acceptance Draft test guideline Submission to regulatory body Consultation with expert groups Adoption and publication of the new test guideline
16
17 THE ECVAM SCIENTIFIC ADVISORY COMMITTEE (ESAC) The ESAC was established by the Commission Communication of October The ESAC advises on ECVAM s work programme, and formally considers the scientific validity of alternative tests. The ESAC members are selected representatives of the Member States, of the European chemical, cosmetic and pharmaceutical industry associations, academic toxicology and animal welfare groups, together with representatives of ECVAM s principal customer DGs. The ESAC meets twice each year, at Ispra.
18 METHODS/STATEMENTS ENDORSED BY THE ESAC - Chemicals 3T3 NRU phototoxicity test November 1997 EPISKIN skin corrosivity test April 1998 Rat TER skin corrosivity test April 1998 Application of the 3T3 NRU phototoxicity May 1998 test to UV filter chemicals Local lymph node assay for skin sensitisation March 2000 EpiDerm skin corrosivity test March 2000 CORROSITEX skin corrosivity test December 2000 Embryonic stem cell test for embryotoxicity May 2002 Whole-embryo culture test for embryotoxicity May 2002 Micromass test for embryotoxicity May 2002 Accepted and incorporated into Annex V and OECD guidelines
19 METHODS/STATEMENTS ENDORSED BY THE ESAC - Biologicals In vitro production of monoclonal antibodies May 1998 ELISA test for batch potency testing of tetanus December 2000 vaccines for human use ToBI test for batch potency testing of tetanus December 2000 vaccines for human use ELISA test for batch potency testing June 2002 of erysipelas vaccines Relevance of the target animal safety test for batch June 2002 safety testing of vaccines for veterinary use Deletion of a test in polycythaemic mice for batch June 2002 potency testing of erythropoietin concentrated solution Accepted and incorporated into European Pharmacopoeia monographs
20 WHAT? European Consensus Platform on Alternatives International Not-For-Profit Organisation in Belgium (Europe) Members : National Consensus Platforms on Alternatives with 4 parties : Academia Industry Government Animal Welfare
21 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS EU WHITE PAPER STRATEGY FOR A FUTURE CHEMICALS POLICY EINECS old substances ELINCS 3000 NEW substances > 95%??? REACH Registration, Evaluation, Autorisation, with PRIORITIES chemicals million animals
22 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS EU WHITE PAPER STRATEGY FOR A FUTURE CHEMICALS POLICY HIGH NEED FOR 3R-ALTERNATIVES million animals
23 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS 7 th AMENDMENT OF EU COSMETICS LEGISLATION ANIMAL TESTING BAN MARKETING BAN OF COSMETIC INGREDIENTS & FINAL PRODUCTS TESTED ON ANIMALS HIGH NEED FOR 3R-ALTERNATIVES
24 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS ECVAM STRATEGIC PLAN FOR FUTURE DEVELOPMENT AND VALIDATION OF ALTERNATIVE METHODS WITHIN NEXT 5 YRS ALTERNATIVES FOR MOST ENDPOINTS HIGH NEED FOR 3R-ALTERNATIVES
25 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS WHAT 3R-ALTERNATIVES DO WE ACTUALLY HAVE IN THE EU FOR REGULATORY TESTING? 4 VALIDATED METHODS ANNEX V Dangerous Substances Legislation 67/548/EEC 3 corrosivity tests TER, Epi Skin,Epiderm 1 phototoxicity test 3T3 NRU-PT
26 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS WHAT 3R-ALTERNATIVES DO WE ACTUALLY HAVE IN THE EU FOR REGULATORY TESTING? 4 VALIDATED METHODS NOT YET TAKEN UP IN EU LEGISLATION 6 METHODS ACCEPTED BY ESAC 1 skin sensitisation test: LLNA 1 in vitro percutaneous absorption test 1 additional corrosivity test: Corrositex 3 embryotoxicity tests: WEC, MM, ECT
27 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS WHAT 3R-ALTERNATIVES DO WE ACTUALLY HAVE IN THE EU FOR REGULATORY TESTING? 4 VALIDATED METHODS 6 METHODS ACCEPTED BY ESAC TESTS UNDER DEVELOPMENT & VALIDATION skin irritation eye irritation acute (oral) toxicity skin corrosion (in silico)
28 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS WHAT 3R-ALTERNATIVES DO WE ACTUALLY HAVE IN THE EU FOR REGULATORY TESTING? 4 VALIDATED METHODS IN VITRO GENOTOXICITY TESTS 6 METHODS ACCEPTED BY ESAC TESTS UNDER DEVELOPMENT & VALIDATION
29 POSITION TOWARDS ACTUAL EU POLICY AND ALTERNATIVE METHODS ALTERNATIVE TESTS NEEDED FOR : PHOTO ALLERGY SUBACUTE TOXICITY CHRONIC TOXICITY REPRODUCTIVE TOXICITY TARGET ORGAN & SYSTEMIC TOXICITY LACKING TODAY BIOKINETICS NON-GENOTOXIC CARCINOGENICITY
30 Præsentationen er udarbejdet med bidrag fra Marlies Halder, ECVAM og Vera Rogiers, ECOPA Endvidere er der anvendt billeder fra og Uffe Midtgård, Leif Simonsen, Lisbeth E. Knudsen: Tksikologi i arbejdsmiljøet, bind II
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