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Rob Taalman

Cosmetics Europe LRSS Programme 2016-20

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Objectives of CE Research

To deliver tools & strategies for animal-free safety assessment

Accurate

Robust

Efficient

…ultimately accepted by regulators…

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Our Systematic Rationale

Exposure

• Exposure upon use

Local effects

• Skin, eye irritation

Penetration, Metabolism, Excretion

• Skin penetration, metabolism

• Bioavailability at systemic target site

Systemic Effects

• Sensitization, genotoxicity

• (Sub-) chronic toxicity

Safety Assessment

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Current LRSS Situation

LRSS = triple package

❶Develop non-animal methods & testing strategies to be used for safety assessment of cosmetics ingredients / chemicals

❷Show that safety assessment – thus probabilistic & realistic assessment – is possible on a broad spectrum of effects / likelihoods including systemic toxicity

❸Advocate for the regulatory acceptance of data generated with such alternative methods

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Current LRSS Situation

5 task forces

1. Eye Irritation

2. Genotoxicity

3. Skin tolerance

4. Bioavailability & Metabolism / ToxicoKinetics (TK)

5. Mode of Action / ToxicoDynamics (TD)

Well advanced combination of

alternative methods towards reg.

accep. Systemic

toxicity…

…based on

non-animal

methods only

Discussed for

decades

Many attempts

to achieve this

But so far not

accepted

Big

challenge

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AAT Strategic Roadmap

2013

Skin Irritation Eye Irritation Genotoxicity

Acute Toxicity

2016

Eye Irritation Genotoxicity

Skin sensitization

Exposure / ADME

Systemic Toxicity

2020

Sensitization

Systemic Toxicity

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Eye Irritation

Eye Irritation Strategy 2012

Eye Irritation Today

()

X

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Genotoxicity

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Evolution of Genotoxicity Requirements

Ames

MLA

MN

Ames

MN

High Sensitivity

Low false

positive ratio

Reduced costs

2003 2004 2014

3D Comet

3D MN

SCCS Notes of guidance – in vitro battery:

Skin assays

accepted follow-

up option

Successful Validation is prerequisite for maintaining status quo!

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Genotoxicity

Strategy supported by SCCS and OECD

Drastically less false positives by 2-assay strategy

Evaluation of tier 2 assays progressing Finalize validation of 3D skin model assays in 2016

Support validation of Hen’s Egg micronucleus test as viable option to follow-up on orally exposed ingredients

Goal to have 2 assays/strategy approved by scientific/regulatory community in 2017/18

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Skin Sensitization

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Metabolism

Immunology

Chemistry

Penetration

Chemical reactivity

Protein modification

Cellular response

Organ response

In vivo response

Integrated assessment

Expert systems

in silico tools

Skin Sensitization - OECD “Adverse Outcome Pathways“ -

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Metabolism

Immunology

Chemistry

Penetration

Chemical reactivity

Protein modification

Cellular response

Organ response

In vivo response

Three in vitro assays validated DPRA, KeratinoSens, hCLAT

Additional 13 assays evaluated Six methods prioritized (incl. DPRA, KeratinoSens and hCLAT)

Promising prediction models being assessed e.g. Support Vector Machines, Bayesian models …

Skin Sensitization - OECD “Adverse Outcome Pathways“ -

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Metabolism

Immunology

Chemistry

Penetration

Chemical reactivity

Protein modification

Cellular response

Organ response

In vivo response

Researching

assays

Skin Sensitization - OECD “Adverse Outcome Pathways“ -

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Overview Skin Sensitization

Achievements

Assays developed / validated / evaluated

Clear roadmap

Status

6 OECD-submitted data interpretation procedures (DIP‘s) evaluated

Assay optimization & data integration

Characterize & expand applicability domain

Challenges (2017-2020)

Potency determination

Testing strategy finalization

Regulatory acceptance

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Systemic Toxicity

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Back to the basics

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Paradigm shift

*adapted from P. Price (US EPA), “Using in vitro data in quantitative risk assessments”

QIVI

VE

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Strategy Systemic Toxicity

Hazard Identification

- Toxicodynamics -

Mechanistic predictions

Exposure

- Toxicokinetics -

Systemic exposure predictions

Safety Assessment

Combined evaluation & extrapolation

Fusion • Concentration response data

• Modelling

- Case Studies -

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Systemic Toxicity - Status

• SEURAT-1 final event 4 Dec. 2015

• SEURAT-1 level 3 case studies Proof-of-concept, TTC, Read-across framework, ab initio

• Differential planning vs. public programs

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Toxicokinetics / ADME - Objectives

Internal exposure

• For read-across and

ab initio risk assessments

• Enable internal TTCs

Guidance for in vitro assays

(concentrations, target organs)

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Toxicokinetics / ADME - Principles

• Topical exposure

• Local concentrations in different organs

• Metabolism

• Distribution

• Excretion

• Organ-specific toxicity

• Reactive metabolite formation or detoxification

• PK profile, Cmax, AUC

Fate of a chemical after exposure via different routes

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Toxicokinetics / ADME - Status

In silico models

Partition/ diffusion

coefficients

Lipid binding

Solubility

Skin penetration (non-viable skin)

Metabolism in ex vivo

skin

Metabolism, stability in

S9

Protein binding

(Physico-) chemical properties

In silico models

Bioavailability & Metabolism

• 50 compounds tested Q3/2016

• In silico models 2017

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Toxicokinetics / ADME - Way Forward

Internal TTC

PBPK ADME Model

Read across / ab initio concepts

In silico

Skin Penetration

Prediction Models

ADME

in vitro Toolbox • intestinal absorption

• liver metabolism

• in vitro biokinetics

• plasma protein binding

Static & Dynamic

3D Skin & Liver

Models

Exposure

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Toxicodynamics - Objectives • Discriminate between specific and non

specific adversities and low toxicity

• Determine perturbed biological

pathways for repeat dose toxicity

• Establish quantitative points of

departure for use in risk assessment

(linked to kinetics)

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Mode of Action

Ontology (2016-2019)

Bio-/ Chemo-

informatics Data (2016-2019)

Toxicogenomics (2017-2019)

Toxicodynamic

AssayToolbox (2016-2020)

Toxicodynamics Building Blocks

Quantify dose which can

alter biological pathway(s)

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Toxicodynamics Building Blocks

Mode of Action

Ontology

(2016-2019)

Bio-/ Chemo-

informatics Data

(2016-2019)

Toxicogenomics

(2017-2019)

Toxicodynamic

AssayToolbox (2016-2020)

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Toxicodynamics Building Blocks - Mode of Action Ontology -

Mode of Action

Ontology

(2016-2019)

Bio-/ Chemo-

informatics Data

(2016-2019)

Toxicogenomics

(2017-2019)

Toxicodynamic

AssayToolbox (2016-2020)

Knowledge-based grouping of chemical features associated with repeat dose toxicity into categories, tie to a putative mode of toxicity (or not)

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Toxicodynamics Building Blocks - Toxicogenomics -

Mode of Action

Ontology

(2016-2019)

Bio-/ Chemo-

informatics Data

(2016-2019)

Toxicogenomics

(2017-2019)

Toxicodynamic

AssayToolbox (2016-2020)

Front line screen for broad characterization of hazard

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Toxicodynamics Building Blocks - Toxicodynamic Assay Toolbox -

Mode of Action

Ontology

(2016-2019)

Bio-/ Chemo-

informatics Data

(2016-2019)

Toxicogenomics

(2017-2019)

Toxicodynamic

Assay Toolbox (2016-2020)

In vitro assays for generating toxicodynamic data Address identified gaps Source of targeting testing

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Toxicodynamics Building Blocks - Bio-/ Chemo-informatics Data -

Mode of Action

Ontology

(2016-2019)

Bio-/ Chemo-

informatics Data

(2016-2019)

Toxicogenomics

(2017-2019)

Toxicodynamic

AssayToolbox (2016-2020)

Informatics and integration/visualization needs of toxicity and ADME data for analysis via case studies

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Fusion & Safety Assessment - Case Studies / Modelling / Evaluation -

Case Studies as Guiding Principle

• Concentration response data / modelling

• Quantitative in vitro-in vivo extrapolation

(QIVIVE)

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1. IDENTIFY USE SCENARIO

TIER 2: APPLICATION

TIER 0: IDENTIFY USE SCENARIO, CHEMICAL OF

CONCERN AND COLLECT EXISTING

INFORMATION

TIER 1: HYPOTHESIS

FORMULATION

2. IDENTIFY MOLECULAR STRUCTURE

3. COLLECT EXISTING DATA

4. IDENTIFY ANALOGUES, SUITABILITY ASSESSMENT AND EXITING DATA

5. SYSTEMIC BIOAVAILABILITY (PARENT VS. METABOLITE(S), TARGET ORGANS, INTERNAL CONCENTRATION)

6. MOA HYPOTHESIS GENERATION (WEIGHT OF EVIDENCE BASED ON AVAILABLE TOOLS)

7A. TARGETED TESTING 7B. BIOKINETIC REFINEMENT (IN

VIVO CLEARANCE, POPULATION, IN VITRO STABILITY, PARTITION)

8. POINTS OF DEPARTURE, IN VITRO IN VIVO EXTRAPOLATION, UNCERTAINTY ESTIMATION, MARGIN OF SAFETY

9. FINAL RISK ASSESSMENT OR SUMMARY ON INSUFFICIENT INFORMATION APPROACH

EXIT TTC

EXIT INTERNAL

TTC

EXIT AB INITIO

EXIT READ ACROSS

(Adapted from Berggren et al. 2016. Publication in preparation)

Systemic Toxicity Workflow

7A. TARGETED TESTING 7B. BIOKINETIC REFINEMENT (IN

VIVO CLEARANCE, POPULATION, IN VITRO STABILITY, PARTITION)

8. POINTS OF DEPARTURE, IN VITRO IN VIVO EXTRAPOLATION, UNCERTAINTY ESTIMATION, MARGIN OF SAFETY

9. FINAL RISK ASSESSMENT OR SUMMARY ON INSUFFICIENT INFORMATION APPROACH EXIT AB INITIO

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It’s all connected

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SA Paradigm = the common matrix to TK and TD TFs

There are some

direct links easy to

draw

And some others

less obvious

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Conclusions – Highlights since 2015

• OECD acceptance of eye irritation RhT tests

• OECD/SCCS acceptance of genotoxicity strategy

• SEURAT-1 case studies

– SCCS: positive reception of TTC for systemic toxicity

– ECHA: positive reception of NAMs (as supporting evidence in read-across)

• Cosmetics Europe LRSS roadmap largely planned

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AAT Means Collaboration !

Industry

- Cosmetics (LRSS)

- Pharma (IMI)

- Life Science

- Chemicals (LRI)

- ECETOC

Public Sector

- E.C. (Horizon 2020)

- JRC

- Academia, institutes

Regulators

- OECD

- ECVAM

- SCCS

EPAA

EPA

Thank you !