TT21C and Safety Assessment at Unilever

Julia Fentem, PhD FBTSVice-PresidentUnilever - Safety & Environmental Assurance Centre (SEAC)

Overview

• Safety Risk Assessment in Unilever

• Non-Animal Approaches for Consumer Safety Risk Assessment

• new technologies & risk-based approaches (exposure)

• Toxicology in the 21C (“TT21C”)• evolving (eco)toxicological risk assessment approaches for

better human health (& environmental) protection

SEAC’s Role

● Risks for consumers, workers and environment– Safety of products and supply chain technology

● Environmental impacts– Sustainability of Unilever’s brands, products & supply

chain

Provide authoritative scientific evidence and expertise so that Unilever can identify and manage:

SAFE and SUSTAINABLEDESIGN and EXECUTION ofInnovation and Technology

Characterise Hazards & Exposure, assess & manage COE Safety Risks across the Value Chain

C = Consumer safetyO = Occupational safetyE = Environmental safety

DisposalRaw materials/ ingredients

Product formulations Manufactureprocess

Transport

Consumer use

formal post-launch monitoring (if warranted)on-going monitoring & review of new data

COE safety risk assessment for market→ safety risk management decision

safety prognosis / identify key risks & datasafe by design considerations

safety risk assessment for clinical / consumer stud ies

O & E exposure scenarios

COE hazards / key safety risks

Integrated Approach to Assessing & Managing Safety Risks

C & E exposure scenarios

SAFE and SUSTAINABLEDESIGN and EXECUTION ofInnovation and Technology

Research into Alternative Non-Animal Approaches

• R&D in-house and with external partners

• work with all key stakeholders

• present & publish our scientific results

• participate in international validation studies

• since 2004 additional €3M per annum investment in:

• innovative research programme on novel consumer safety risk assessment approaches without animal testing

• new technical capabilities, e.g. “systems biology”, omics technologies� evaluating applicability for decision making on safety risks

• working with >50 external partners globally

• output shared

Embracing New Technologies

How can our consumer safety risk assessments benefit from applying new & emerging technologies being used in medical and biological research?

• “omics”• genomics, proteomics, metabolomics …• tools to interrogate biological systems at molecular level

• informatics• computational and mathematical approaches• tools to integrate, analyse, visualise and interpret data

• analytical• advanced chemical and immunological detection methods

• bioengineering• tissue constructs, stem cells …

Consumer Safety Risk Assessment

Computational Chemistry

Stem cells

US ‘Human Toxicology Project’ consortium

Advanced organ-simulating devices

Human-based cells in vitro

Computational modelling

Systems Biology and PBPK modelling

Tissue Engineering

Novel Approaches to Risk Assessment

Out-reach to Regulators

Mapping and modelling cellular circuitry controlling toxicity pathways

Pathway assays

Relationships between pathway perturbations and adverse responses

Human health risk assessments

Working with Others on New Technologies & Risk -Based Approaches

● technical collaboration with leading US scientists

– Hamner Institute, North Carolina– implement US NAS strategy (2007)

– commonalities with research strategy Unilever published in 2004

● engaging with Russian & Dubai government authoritie s– safety risk assessment approaches for consumer products– inclusion of non-animal methods in revised technical regulations

● collaborating with Chinese Academy of Sciences / La ncaster Uni., UK– environmental risk assessment of chemicals

Developing & Sharing our SafetyApproaches with Scientists Globally

EU: SCCS Opinion 2010

‘…. majority of the existing alternative methods [are] only suitable for hazard identification of cosmetic ingredients and do not give information on potency. Thus, a full human health riskassessment cannot be performed’

http://ec.europa.eu/health/scientific_committees/consumer_safety/statements/index_en.htm

Alternatives to Animal Testing (Safety Decisions)current scientific reality – 2010 expert report (EU Commission)

Source: EU Commission ‘Report on Alternative (Non-A nimal) Methods for Cosmetic Testing: Current Status and Future Prospec ts – 2010’

Human HealthToxicology Endpoint(subject to 2013 ban)

Timeline to Replace Animal Testing

[Note: regulatory acceptance would require an additional 4-8 years]

Comments

Repeated dose toxicity “no timeline could be foreseen” projects still at early research stage

(EU Comm / COLIPA partnership - SEURAT)

Carcinogenicity “no timeline could be foreseen”current in vitro tests inadequate for

generating dose-response information required for safety assessment

Skin Sensitisation “2017 – 2019 for full replacement”several non-animal approaches under

evaluation; ways to integrate & interpret data for safety assessment required

Reproductive Toxicity “no timeline could be foreseen”projects still at research stage

(EU- & US-funded programmes)

Toxicokinetics “no timeline could be foreseen”projects still at research stage

(focus for US “TT21C” research)

US NRC Report

“Advances in toxicogenomics, bioinformatics, systems biology, epigenetics, and computational toxicology could transform toxicity testing from a system based on whole-animal testing to one founded primarily on in vitro methods that evaluate changes in biologic processes using cells, cell lines, or cellular components, preferably of human origin.”

“TT21C” – Evolving Toxicological Risk Assessment → Better Safety Decisions “Toxicology in the 21C”

– modernisation agenda – developing & applying latest advances in S&T; transforming the toxicological hazard / risk paradigm through better understanding & interpreting effects at the cellular & molecular levels

– new scientific understanding & approaches will underpin future regulatory changes in chemical safety risk management approaches; need scientifically robust ways to integrate data from various tests & analyses

Oxidative stress response module

Developing new S&T Capability for Risk and Impact Assessments – SEAC Focus

Environmental Sustainability• Greenhouse Gas Footprint• Water Footprint

Non-Animal Approaches for Consumer Safety Risk Assessment• Skin Allergy• Exploring Risk Assessment – TT21C• New Technologies• Applying Non-Animal RA Approaches

Ecotoxicology Risk Assessment• Improving Higher Tier RAs• Improving RAs in India & China• Screening RA MethodsMicrobiological Safety Risk Assessment• Naturals & Naturalness RA• Functional Biological Agents• Water (Devices, Treatments & Ecology)

Safe and Sustainable Process Technology• Exposure Risks for Novel Ingredients• Sustainability by Design

Toxicology Risk Assessment• Functional Actives• Protein Allergy• Exposure

Chemistry• Complex Mixtures & Naturals• Proteins & Enzymes• Bioanalysis & Systemic Exposure

Cross-Domain• Risk Assessment of Nanoparticles

Non-Animal Approaches for Skin Allergy

• Current hypothesis: several non-animal hazard characterisation approaches required as inputs into risk assessments of the future

• e.g. Maxwell et al (2008) ATLA, 36, 557-568

• Understanding mechanistic basis of skin sensitisation using a mathematical modelling / systems biology approach

• This has guided selection & development of in silico and in vitroapproaches that encompass key events in Skin Sensitisation induction

• Chemical reactivity

• Peptide reactivity

• Skin disposition

• Skin bioavailability

• Skin inflammation

• Dendritic cell activation/maturation

• T cell proliferation

• Basic research still required in some areas to fill gaps in understanding

Induction Elicitation

Dendritic cell activation and migration

Calculationof net

proliferation

Chemicalexposure

Epidermal cellactivation

Dendritic cell presentation of antigen and T cell proliferation

Maxwell G. & MacKay C (2008) ATLA 36, 521-56

Modelling the Underlying Pathways

Components of the TT21C Vision

The Future of Risk Assessment?

Fast high-content information in vitro assays in human cells/models

Dose-response assessments

Computational models of the circuitry of relevant t oxicity pathways

Pharmacokinetic models supporting in vitro to in vivo extrapolations

Risk assessment focused on maintaining exposures be low the levels that significant pathway perturbations occur

CONSUMER EXPOSURE SCENARIO(S)

Biologic

Inputs

Normal

Biologic

Function

Morbidity

and

Mortality

Cell

Injury

Adaptive Stress

Responses

Early Cellular

Changes

Exposure

Tissue Dose

Biologic Interaction

Perturbation

Based on Perturbation of Toxicity Pathways

1001010.10.01

1

0.9

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0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

dose

Res

pons

e

i. In vitro rapidly performed toxicity pathway

test battery for n-assays in human cells , cell

lines, or tissue aggregates

ii. Computational systems biology description of

pathway circuitry for creating biologically

realistic dose response models

iii. Dose dependent transition studies for

sequential pathway activation to understand

linkage to cell and tissue level responses

(perturbations to adversity)

iv. PBPK Modules – Compound specific or QSAR-

based models for reverse dosimetry based on

adverse concentration defined in the in vitro

studies

Toxicity Pathway Results and Quantitative Risk Assessments

In Vitro to In Vivo (human exposure) Extrapolation

Exposure mg/kg/day

Target site concentration ( µM)

In vitroadaptive/adverse

threshold concentration (µM)

TT21C – Main Players

Toxicity Testing in the 21 st Century (TT21C)

Prototype Pathway Research for TT21C –Hamner/Unilever Partnership

A TT21C Prototype Toxicity Pathway: Thresholds for Genotoxic Activity

– Traditionally risk assessments of ‘genotoxins’ have been based on linear models

– At low doses, mechanisms prevent damage from becoming a permanent defect

– Defence mechanisms reach saturation; departure from the NOAEL

– Critical exposure level, below which the concentration of a compound will not produce a significant increase in mutation or chromosomal effects

Background

Dose of GenotoxinMutation

Background

Dose of GenotoxinMutation

Background

Dose of GenotoxinMutation

(Jenkins, et al., 2009, Toxicology)

Can we use mutagenicity and clastogenicity tox-path ways in TT21C paradigm to

(a) improve Genetic Toxicology RA (no animals) and

(b) provide a prototype “reason to believe” (proof of principle) for TT21C ?

• Develop tools to assess dose response for DNA-damage stress pathways, linking HCA, gene expression and mutation, to assess dose-dependent transitions for case-study mutagenic compounds

• Applying data to develop a computational systems biology model of the p53-mdm2 network to examine the basis of the dose-dependent transitions in mutational efficiency� providing a TT21C risk assessment approach

Joint Research Programme

Joint Research Programme

• Protein response to DNA damage• phos-p53, total-p53, p21, MDM2, Chk2, p-ATM, H2AX• Localization of Mn & DNA repair proteins in single cells

• Alterations in gene expression following DNA damage• Time and dose-dependent changes • Full-genome arrays

• Computational modeling of dose response for DNA damage pathway activation

Integrate Data into Models

� Basal function

� Response to small

perturbations

� Response to larger

perturbations

� Assess threshold level of

damage to increase

mutation

Some Challenges ...

Scientific

• Increase collaborative research (EU – US – China ....) – common roadmap(s)

• New research ideas linked to understanding pathways & outcomes critical for human health and environmental risk assessments

Key Question: is use of ingredient X at concentration Y in product Z safe –

for consumers?– for the environment?

Some More Challenges ...

Regulatory Acceptance• Traditionally, in vitro alternative tests validated as 1 for 1 replacements for

existing animal tests• ECVAM / ICCVAM validation

• OECD guidelines

• Regulatory acceptance

• Likely that a pathways-based approach will involve a ‘toolbox’ of several different non animal-based methodologies, none of which (by themselves) will be a ‘replacement’ for a current animal test

• Tests based on understanding toxicity pathways for use in risk assessments will need a new approach to gaining acceptance

• Understanding robustness and transferability of the tests themselves

• Acceptance of new paradigms for use of new data in risk assessments

2007-2008: education, discussion of

content and rational

2009-2010: synthesis and discussion of issues

and challenges

Into the future: moving forward with

implementation – a pilot project approach

Implementation of

TT21C for Testing

and Risk

Assessment

Profiling &

Prioritizing

with HTS

Testing

Systems

Toxicology for

Pathway

Identification

and New

Methods

Pilot Projects

emphasizing

application to

risk assessment

Different Opportunities for Implementation

And, others –

public-private

partnerships

The TT21C

Report

Exposuretissue

dose

Apical

Endpoint

(cancer)

intermediate

responses

“likely to be

a high dose

toxicant”

concentration

in vitro

pathway

assays

Evaluation of ‘adverse’ degree of

system perturbation from panel of

assays and computational systems

for d-r modeling

Value of Prototypes

Mode of Action

Framework

TT21C

Approaches

p53-mdm-2 DNA damage pathway –

Unilever & Exxon-Mobil

PPAR-α α α α receptor pathway mapping and

modeling - Dow Chemical

Rat/human tissue surrogates, in vitro-

in vivo extrapolation and genomics –

ACC

Oxidative stress pathway – Dow

Chemical and Sumitomo

Extend on-going

work

Expand group of

pathways

Develop training

component

Plan use in

safety/risk

assessment

Status

http://www.tt21c.org

With special thanks to ...

• Mel Andersen• Director, Institute for Chemical Safety Sciences, Hamner Institutes

• Paul Carmichael• Cameron MacKay• Gavin Maxwell• Carl Westmoreland