Overview of the Latest Scientific Developments on Animal Models and Alternatives

Dr Philip A Botham Global Head of Human Safety Syngenta

Animal Models for What ?

Its not just about toxicology / safety evaluation

Statistics of Scientific Procedures on Living Animals : Great Britain 2000

Total number of Procedures Total number of animals used 2,714,726 2,642,993

Total number of toxicology procedures 454,904 (16.8%)

Total

number of toxicology procedures for Cosmetics and Toiletries Total number of acute lethal tests in the rat (LD50/LC50)

0 2292

Total number of Procedures in Dogs Total number of procedures in primates

7,632 (0.3%) 3,690 (0.1%)

The vast majority of animals used in the European Union are used in:
Fundamental biological research Applied research in human medicine and dentistry Applied research in veterinary medicine Breeding programmes, especially for genetically modified animals

The Use of Animals in Toxicology Mandatory Tests

For all industry sectors (pharmaceuticals, agrochemicals, industrial chemicals, cosmetics, household products)
Regulatory requirements for conducting clinical trials, registration, safe manufacture, transport Assess hazard not risk Wide range of complex hazard endpoints (acute, chronic, reproductive system, cancer, teratogens, sensitisers) New endpoints / tests (nervous system, immune function, susceptible human sub-populations, e.g. children)

The Use of Animals in Toxicology - Elective Tests

Compound selection in discovery (active ingredient) and formulation (product) development Mechanistic studies are findings seen in mandatory tests in rodents / dogs relevant to man? Research studies understanding generic mechanisms, e.g. how certain chemical classes cause cancer

Future Trends in Laboratory Animal Use in Toxicology

New toxic endpoints more testing More products of biotechnology / fewer traditional chemicals change in testing requirements Greater emphasis on understanding mechanism of toxicity
Relevance of animal models Transgenics (humanisation of models) Use of in vitro and in silico technology

Public and regulatory expectation for safer drugs pesticides, chemicals and food and for reduced animal use.

Toxicology offers both a threat and an opportunity for reduction, refinement and replacement alternatives to animal experimentation

What Progress Has Been Made ?

Replacement
Skin corrosion Phototoxicity Skin permeability

Refinement / Reduction
Acute oral toxicity Skin sensitisation
INVENTION TO REGULATORY ACCEPTANCE TOOK 15 20 YEARS

What Tests Are in Development or Validation?

Available within 3 years
Acute oral toxicity (in vitro screening/dose-setting for in vivo studies) Skin irritation (in vitro - replacement) Developmental toxicity (in vitro-screening for moderate to strong teratogens)

Available within 5 10 years

Eye irritation (in vitro replacement) Acute dermal and inhalation toxicity (in vivo refinement / reduction) Respiratory sensitisation (in vivo new endpoint)

In-Vitro Replacement Tests Available Only in Longer Term (more than 10 years)
Acute toxicity Skin and respiratory sensitisation Kinetics and metabolism Target organ / system toxicity Developmental and reproductive toxicity Non-genotoxic carcinogenesis

Is this pessimistic or too conservative ? Why does it take so long to develop and validate alternatives ? Would more investment in alternatives speed up progress ?

Stages in the Development of New Toxicology Test Methods

Stage
Research Method Development Pre Validation Validation Review Regulatory Acceptance

Outcome
Understanding basic toxic or biological mechanisms New methods for specific applications Optimised transferable protocol Establishment of reliability and relevance Independent scientific peer review Regulatory agency decision on acceptability for specific application

Science or Technology
Science Science

Technology Technology -

Technology can be driven to a timetable by the application of sufficient resources and management skills, while science has a pace of its own
IFH Purchase, 1996

Regulatory acceptance also has a pace of its own, and is perceived to be driven as much by politics as by science
PA Botham, 2002

I know, lets invent the Ames test

Would More Investment in Alternatives Speed up Progress ?

Yes by giving more support for conduct and management of high quality validation studies Yes by encouraging test developers to better understand the needs for hazard and risk assessment in toxicology (more collaboration between academics and industry) No by funding poorly conceived or poorly conducted test development

Test Development A Current Example of Expectation Not Matching Reality

Genomics, Transcriptomics, Proteomics and Metabonomics

Genomics, Transcriptomics, Proteomics and Metabonomics (GTPM)

Mechanistic toxicology; improve relevance to man Predictive toxicology; biomarkers for particular toxic endpoints or classes of toxiciant Can be used with in vitro culture systems

The Problems with the Development of GTMP Technology in Toxicology

e.g. Using transcriptomics (toxicogenomics) Changes seen genuine adverse effects or healthy adaptive or repair responses? Effects seen at very low doses (relevance ?) Generic problems with the use of in vitro culture systems
Transcript changes may not reflect what would happen in an organ in vivo

Different microenvironment Lack of cellular interactions Inadequate or inappropriate metabolism

The appropriate application of these techniques is more demanding of careful experimental design than ever, as the potential to generate incomplete and misleading data is great. The attainment of common ground through collaboration involving the generation, sharing and publication of suitable, high quality, data should be prime goal for scientists and institutions engaged in researching the new technology and its appropriate application towards improving the knowledge of the interaction of chemicals with living things.
B Pennie, 2001