EUROPE GOES ALTERNATIVE Animal Testing and Alternative Methods - a Scientific Perspective Coenraad...

‘EUROPE GOES ALTERNATIVE’

Animal Testing and Alternative Methods -

a Scientific Perspective

Coenraad F.M.Hendriksen

Netherlands Centre Alternatives to Animal Use (NCA), Utrecht University

& Netherlands Vaccine Institute (NVI)

Animal Testing and Alternative Methods - a scientific perspective

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Conference ‘Europe Goes Alternative’, Brussels, 071105

Topics of presentation

science and alternatives -

the way to go!

Other reasons for going

alternative

3Rs recommendations

Conclusion

Science and Alternatives - the way to go!

phenotypeTreatment Incubation time

minutes years

Until now questions are based on ‘what’ :

- what is the toxicity of a chemical?

- what is the potency of a vaccine?

- what is the safety of a pharmaceutical?

Future: ‘why’ questions will dominate:

- why is a chemical toxic?

- why is a batch of vaccine potent?

- why is a pharmaceutical not safe?

Scientific and regulatory evolution: from ‘what’ to ‘why’ (1)

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(Timo Breit)

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Paradigm shift in Science and regulatory testing: from ‘what’ to ‘why’ (2)

‘What’ questions

Parameter Endpoint (animal model)

vaccine potency protection after challenge

chemical toxicity increase tumour frequency

drug safety clinical signs


‘Why’ questions

Endpoint Model (e.g.)

3-D configuration antigen physiochemical

DNA or DNA repair change

at molecular level tissue culture

interaction at receptor level tissue culture


New developments

Examples - omics technologies & bioinformatics embryonic stem cells in silico modelling tools improvements in tissue culture techniques systems biology etc.

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Micro-array analysis:1.000-40.000 genes




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Paradigm shift in science and regulatory testing: from ‘what’ to ‘why’ (3)

Conclusion:

Future will give a shift in research and regulatory testing paradigms: non-animal models will increasingly being used to generate answers to the ‘why’ questions (mechanistic info) while the animal model will be used to verify and confirm data from non-animal studies


Past/’present’ ‘Future’

R & D start

End of research objective

Other reasons for Europe to go alternative

Animal use & concerns

Moral concerns

Economic and pragmatic concerns

Scientific concerns

(In)validation concerns

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Moral concerns


Scientific concerns


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%

Societies acceptance of animal use for specific purposes

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cosmetics household productspharmaceuticals AIDS vaccineleukaemia child

(Aldhous et al. 1999)


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Increased legislation and regulation concerning laboratory animal use

Revision of Council

Directive 86/609/EEC

Appendix A of the

European Convention


Moral concerns


Scientific concerns



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Moral concerns


Scientific concerns


Europe goes alternative -

Future trend in Lab. Animal use?

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year year

Europe goes Alternative: the way forward

Objective Conference

demonstrate that the European Union keeps animal welfare high on the political agenda and remains in the lead for animal protection,

show industry’s approach to actively promote research in alternative methods,

illustrate progress made in the area of alternative methods to animal tests at EU and intermational level,

identify further possibilities to improve development, validation and legal acceptance of alternative methods

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R eality

ealism

eactionR

Rex Burch Bill Russell

R

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Objective: identify further possibilities to improve development, validation and legal acceptance of alternative methods

Effective EU policy towards

‘Europe goes Alternative’

should include the following elements

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a b c d e Tox.&Safety

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tox.&safetytestingcosmetics

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Focus of Commission concerning 3Rs research priorities

Reality: numbers and purposes of animal use

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Training

Fund. Biology

R&D medicines

Production &q.c. medicines

Diagnosis

Tox. & Safety

Others

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No. of animals used in EU-MS in 2002 (N total = 10.731.020): purposes

Reality: numbers and purposes of animal use

Reality: alternatives and the 3Rs principle

Replacement of animal tests by

methods not using animals

Reduction: reducing the

numbers of animals needed to answer a scientific question

Refinement: reducing pain

and distress in a particular animal experiment

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Reduction:

‘….of all modes of progress it is the one most obviously, immediately, and universally advantageous in terms of efficiency.’

(Russell & Burch, 1959)

Refinement:

‘Happy animals make good science.’

(Trevar Poole)

Replacement:

‘Desirable as replacement is, it would be a mistake to put all our humanitarian eggs in this basket alone.’

(Russell & Burch, 1959)


Reality

Realism

Reaction

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Realism: Proposed timeframe for full replacement of animal tests

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Human Health effects Foreseeable time involved for full replacement

_________________ _____________________

acute toxicity > 2014 skin irritation > 2014 skin corrosion < 2004 eye irritation > 2010 skin sensitisation > 2019 skin absorption/penetration > 2006 subacute/subchronic toxicity >> 2014 (not foreseeable) genotoxicity/mutagenicity > 2016 UV-induced effects >> 2019 (not foreseeable) Toxicokinetics and biotransformation >> 2014 (not foreseeable) Carcinogenicity >> 2014 (not foreseeable) Reproductive toxicity >> 2014 (not foreseeable)

(SCCNFP, 2004)Conference ‘Europe Goes Alternative’, Brussels, 071105

Realism: Legislative and Regulatory context

Sectorial regulation

Regulations to animal testing and multilateral agreements, e.g. Council Directive 86/609/EEC

(According to Schiffelers et.al.2005)

Council Directive

pharmaceuticals

Council Directive

vaccines

Council Directive

pesticides

Council Directive

chemicals

Horizontal legislation

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Examples of conflict between sectorial European directives and EU legislation 86/609/EEC:

re-testing of biological products entering the European market

shellfish toxin testing policies

Other concerns

Slow progress in Harmonisation and/or mutual acceptance

Increased priority for risk minimisation

Sectorial regulations and decisions regulate

the activities of particular sector.

Horizontal legislation interacts with

aspects of sectorial regulations


Reality

Realism

Reaction

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Reaction: obstacles concerning implementation of validated alternatives

Council Directive 86/609/EEC on the protection of laboratory animals

Art. 7 (2)An experiment shall not be performed if

another scientifically satisfactory method of obtaining the results sought, not entailing the use of an animal, is reasonably and practically available

Art. 7 (3)In a choice between experiments, those

which use the minimum number of animals, cause the least pain, suffering, distress or lasting harm and which are most likely to provide satisfactory results shall be selected.

Examples:

- European Pharmacopoeia monographs for Diphtheria and Tetanus vaccine specify two possibilities for potency testing, substantially differing in the level of suffering from each other

- Production of monoclonal antibodies

Conference ‘Europe Goes Alternative’, Brussels, 071105NC

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se The difficulty lies not in the new ideas, but in escaping from

the old ones(John Maynard Keynes, 1883-

1946)

Europe goes Alternative: the way forward

Objective Conference

demonstrate that the European Union keeps animal welfare high on the political agenda and remains in the lead for animal protection,

show industry’s approach to actively promote research in alternative methods,

illustrate progress made in the area of alternative methods to animal tests at EU and international level,

identify further possibilities to improve development, validation and legal acceptance of alternative methods

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Alternatives developed and validated for regulatory testing in the last decade


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Chemicals/biologicals

Animal model Alternative model

skin corrosivity test tissue culture tests phototoxicity tests cell culture test embryotoxicity tests cell culture test skin sensitization test LLNA test potency testing of tetanus Elisa test and ToBI test

potency test erysipelas Elisa test

vaccine target animal safety test deletion monoclonal antibody in vitro culture

production



Alternatives developed and ‘validated’ for regulatory purposes in the last decade


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skin corrosivity test tissue culture tests phototoxicity tests cell culture test embryotoxicity tests cell culture test skin sensitization test LLNA test potency testing of Elisa test, ToBI test

tetanus vaccine monoclonal antibody in vitro production

production classical LD50 tests 3 alternative approaches pyrogenicity testing LAL test and in vitro model percutaneous adsorption cell culture test production inactivated in vitro culture

polio vaccine (monkeys) abnormal toxicity test test deleted specific tox, diphtheria cell culture test

vaccine potency test diphtheria cell culture test

vaccine potency testing of diph- single dose testing

theria and tetanus vaccine potency test polio vaccine antigen quantification



potency test hepatitis antigen quantification

vaccine neurovirulence test PCR & transgenic animals

polio vaccine potency test erysipelas Elisa test

vaccine potency test various clos- Elisa tests

tridial vaccines safety test erysipelas test deleted

vaccine potency test leptospirosis antigen quantification

vaccine target animal safety test safety testing poultry in vitro methods

vaccine abnormal tox.test hormones test deleted safety testing hormones test delted potency test rDNA physico-chemical tests

hormones lethal endpoints use of humane endpoints

Alternatives developed and validated for regulatory purposes in the last decade (tests developed in Europe are given in blue)


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skin corrosivity test tissue culture tests phototoxicity tests cell culture test embryotoxicity tests cell culture test skin sensitization test LLNA test potency testing of Elisa test, ToBI test

tetanus vaccine monoclonal antibody in vitro production

production classical LD50 tests 3 alternative approaches pyrogenicity testing LAL test and in vitro model percutaneous adsorption cell culture test production inactivated in vitro culture

polio vaccine (monkeys) abnormal toxicity test test deleted specific tox, diphtheria cell culture test

vaccine potency test diphtheria cell culture test

vaccine potency testing of diph- single dose testing

theria and tetanus vaccine potency test polio vaccine antigen quantification



potency test hepatitis antigen quantification

vaccine neurovirulence test PCR & transgenic animals

polio vaccine potency test erysipelas Elisa test

vaccine potency test various clos- Elisa tests

tridial vaccines safety test erysipelas test deleted

vaccine potency test leptospirosis antigen quantification

vaccine target animal safety test safety testing poultry in vitro methods

vaccine abnormal tox.test hormones test deleted safety testing hormones test delted potency test rDNA physico-chemical tests

hormones lethal endpoints use of humane endpoints

Europe goes alternative!

Vision without action is daydream~

Action without vision is nightmare

--Japanese proverb

Recommendation: an independent advisory board to the Commission to identify

critical aspects of animal use,

priorities in 3Rs research

potential obstacles concerning implementation, and

potential conflicts between 86/609/EEC and other Directives

Thanks for your attention!!

Validated non-animal tests

Only 9 non animal tests endorsed as scientifically valid by ECVAM:

skin corrosivity (Rat TER, CORROSITEX, Episkin, Epiderm),

phototoxicity (3T3NRU),

embryotoxicity (EST, MM, whole embryo assay),

in vitro percutaneous absorption.

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Annual European Meeting Toxicology Forum, Brussels, 091105

if so, why do we still use animals?

Ontwikkelingen in het gebruik van proefdieren

NCA jubileum symposium, 25 november 2004

(De Boo)

EUROPE GOES ALTERNATIVE Animal Testing and Alternative Methods - a Scientific Perspective Coenraad...

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