Heidi Foth, December 2009

Architecture of REACH

Concept of REACH Obligations & Dynamics Needs for Predictive Testing Alternative Methods
Heidi Foth, Martin Luther Universitt, Halle/Saale Germany
email heidi.foth@medizin.uni-halle.de
2009-11-30 1

Heidi Foth, December 2009

Expectations on REACH
High number of non evaluated chemical substances Generate an information platform without imbalance Existing regulations on chemicals are highly developed but cannot gain speed Omit time consuming discussion on minor risks The power to regulate is not sufficient for some circumstances dispersive use of chemicals with dangerous properties persistence of widely used chemicals impact on environment Obtain an overview on downstream use Complex distribution across the production chain
2009-11-30

Establish a new platform of information

Concept of REACH

Heidi Foth, December 2009

REACH has strong roots

REACH is needed to protect man and environment from damage ..... ? Chemical Safety before REACH What will continue ? ? Concept of REACH What is the strategy for changes ? ? Key elements within REACH Where is the power to save time and resources ?

? Responsibilities
What is rearranged ?
2009-11-30 Concept of REACH 3

Heidi Foth, December 2009

Key elements within REACH

before REACH New notified Substances Special Chemicals Classification: Chem/physical Hazards Toxicity CMR Pharmaceuticals Pesticides, Substances for cosmetics CMR Compounds Persistent organic substances
2009-11-30

under REACH Registration of Chemical Substances all substances >1 t / a Test data Classification plus Risk Evaluation All chemical substances with very dangerous properties Also EINECS listed substances >1 t / a

Evaluation of Dangerous Properties

Authorisation of Production and Use with/without restrictions

of CHemicals CHemicals
Concept of REACH

Heidi Foth, December 2009

Registration Evaluation Authorisation of CHemicals CHemicals

Substances produced / imported

Dangerous properties; Exposure

Production / use with or without restrictions

classification concerning hazards technical use & risks of chemicals health risks for consumers risk for environment

about 30.000 EINECS listed Chemicals, Notified new chemical substances

Close information gaps for industrial chemicals (no data no market) No time consuming discussion on minor risk Approaches to gain speed grouping of substances, predictive tools (QSAR), adaptation of test regimes Need for justification of test on vertebrates Authorization of dangerous chemicals (restrictions in use)
2009-11-30 Concept of REACH 5

Heidi Foth, December 2009

Registration of Chemical Substances under REACH

All substances 1 t/a per Manufacturer or Importer June 1, 2007

Manufacturer/Importer - information on registrant pre-registration - identity of substance (CAS) - compiles and publishes lists PRE REGISTRATION of pre-registered substances - time line for registration June 1 - Dec 1, 2008 - production or import may continue PHASE-IN - Substance Information Exchange Forum Time lines for registration New >1000 t : Dec 1, 2010 substances CMR (1&2): Dec 1, 2010 June 1, 2008 PBT, vPvB: Dec 1, 2010 100 -1000 t: June 1, 2013 New Notified Substance 1 - 100 t: June 1, 2018 ELINCS list (Dec 1, 2008) Manufacturer / Importer Classification, labeling, Agency REGISTRATION Safety data sheet (SDS), registration Chemical safety report (CSR) - registration number for Chemical safety assessment (CSA) New Notified Substances Submission of dossiers - technical evaluation of dossiers Proposal for test on vertebrates - substance evaluation

Agency

Existing Substances EINECS list + other substances

2009-11-30

Concept of REACH

Heidi Foth, December 2009

aim

Protection of downstream users, consumers & environment from damage SDS CSR CSA (**) Guidance of safe use Risk Assessment

Procedure

documents

Exposure scenarios

DNEL, PNEC (*)

Amount produced & production chain

Hazard Classification and Labeling

duty

Registration and Evaluation of products by manufacturers and importers

Concept of REACH 7

2009-11-30

Heidi Foth, December 2009

Obligations for Industry

Registration Evaluation Authorisation of CHemicals

All* Substances produced / Imported 1t/a

Dangerous properties; Exposure Scenarios Classification of hazards Collection of all identified uses Exposure Evaluation of risk(s) Risk management measures Communication of results Regulation (EC)1907/2006

* exemptions

2009-11-30

Obligations & Dynamics

Heidi Foth, December 2009

Important Time Lines

Preregistration * until December 2008 143. 000 Pregistrations (Exclusion) Time line(s) for Registration first group Dec. 1, 2010 substances 1000 t/a; all CMR substances 1 t/a; all R 53 substances 100 t/a June 1, 2013 all substances 100 t/a; June 1, 2018 all substances 1 - 100 t/a find a Registrant in EU

second group third group

Outside EU

* Substance Information Exchange Forum (SIEF)

2009-11-30 Obligations & Dynamics 9

Heidi Foth, December 2009

Expected Workload
Registration of 2700 Substances > 1000 t/a in 3 years
ICCA/OECD 400 Substances in 7 a 1000 Substances in 12 a expected BUA 300 + 220 Substances in 20 a EU 120 from 140 Substances in 12 a MAK 1000 Subst, BG-Chemie 447 Substances

Registration of 4200 Substances 100 1000 t/a in 6 years Registration of 7200 Substances 10 100 t/a in 11 years Registration of 17 500 Substances 1 10 t/a in 11 years Many times more ?
2009-11-30

29.7 % also belong to the higher production volume

33.7 % also belong to the higher production volume

unknown overlap in production volumes

Obligations & Dynamics

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Heidi Foth, December 2009

Approaches to gain speed (1)

Registration needs data from Standard Testing The work load depends on A) the amount produced

B) Expectations on exposure scenarios

Risk Management Measures relieve from work load

Exchange of Information

2009-11-30

Obligations & Dynamics

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Heidi Foth, December 2009

Approaches to gain speed (2)

Standard testing can be adapted Use of existing data, weight of evidence, (Q)SAR, in vitro methods

Testing can be waived Substance tailored Exposure driven Testing

2009-11-30

Obligations & Dynamics

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Heidi Foth, December 2009

Use of Chemicals
Chemicals have useful physico-chemical properties Chemicals are intensively used and widely distributed Chemicals have manyfold biological effects Distinction between wanted and unwanted effects and restrictions for use Drugs Pesticides Compounds for cosmetics Safe handling and prevention of damage for Industrial Chemicals ?
2009-11-30 Needs for predictive Testing 13

Heidi Foth, December 2009

Safe use of Industrial Chemicals

on humans in workplace Production Accidents Regular work Transport Small Trades during use Recognition of Chemicals with carcinogenic, mutagenic and reprotoxic properties Persistent organic substances
Needs for predictive Testing

for the environment long term effects of emission on Water body Air Soil

2009-11-30

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Heidi Foth, December 2009

Acute Toxicity
Empirical Science Mostly in vivo tests on animals Assessment of Pathology Essential endpoints of every study etiology pathogenesis morphology significance Search for life threating effects and for toxicity on CNS, liver, lung, kidney OECD Nummern Rodents Rats Mice

Non Rodents Rabbits Guinea pigs Dogs Pigs Sheep

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2009-11-30

Needs for predictive Testing

Heidi Foth, December 2009

Local effects
Erythema Eczema Hyperkeratosis Exsudative Erythema Bullous Epidermolysis Chemical irritation + immun(toxic) Reaction?

Figure 5. A, B. Light microscopy of HD exposed HGP skin 24h postexposure; microblisters (mb), dermal epidermal junction (dej)
J Med CBR Def | Volume 3, 2005

Figure 6. A , B. Light microscopy of DMSO-HD exposed HGP skin 24h postexposure with exacerbated microblisters (mb) and expansive cleavage at the dermal-epidermal junction (dej)

2009-11-30

Needs for predictive Testing

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Heidi Foth, December 2009

Chronic Toxicity
Reason for difference in response between acute and chronic toxicity Empirical background for repeated dosing / chronic studies
Storage in tissue Induction in metabolism Modified gene expression Adaptive responses apoptose(dys)regulation cell cycle control tumorsuppressor genes immunresponse regulation of hormones adhesion proteins transporter
2009-11-30

metals, persistent organic polutants PAKs, some pharmaceuticals, ROS, gases, PAKs, metals

many carcinogens s.o. s.o. AllergeneS, Immunsuppression xenoestrogenes thalidomid, besides others cytostatics, drugs, some metals
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Needs for predictive Testing

Heidi Foth, December 2009

Target Organs: Bone Marrow

Neutrophil Eosinophil

Myeloic Leucocytes Megacaryocyt Plasmacell Erythron

Differential leukocyte count

2009-11-30

Neutrophils

tissue necrosis, phagocytosis) acute infection, hepatitis) inflammation mediator, hypersensitivity) allergy, poisoning)

Lymphocytes ( Basophils Eosinophils Monocytes ( (

( poisoning)
Needs for predictive Testing 18

Heidi Foth, December 2009

Target Organs: Liver

Acute Effects Dimethylnitrosamine Ethylenchlorhydrine yellow phosphous dystrophy necrosis steatosis fibrosis cholestasis Halogenated aliphatic hydrocarbons Ethanol cirrhosis primary liver cell carcinoma angiosarkoma Chronic effects Methylendianiline -Naphthylisothiocyanate Arsen, Paraquat Arsenite Thoriumdioxid, Vinylchlorid, peroxisomal proliferators Tetrachlormethane Chlorinated aliphatic hydrocarbons

Energy metabolism, endocrine function, vitamin & trace elements Phase I and II metabolism of xenobiotics
2009-11-30 Needs for predictive Testing 19

Heidi Foth, December 2009

Target Organs: Kidney

Kidney toxicity

Kidney tumor

Excretion of waste substances Regulation of liquid volume Water-electrolyte balance Acid-base balance Production of hormones

2009-11-30

Needs for predictive Testing

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Heidi Foth, December 2009

Target Organs: Lung

Bronchitis COPD Emphysema NOx, SO2, O3 flour dust, Sprays, wood dust ? Exogenous factors ? protease excess, 1-antitrypsin deficit Metal dust, Paraquat Bleomycine, Amiodarone PAH, TSNA, Mineral fibres

Dyscrinia Obstruction Respiratory Insufficience Damage Tumors

Interstitial Disease Cancer

Gas exchange
2009-11-30

Blood pH regulation
Needs for predictive Testing 21

Heidi Foth, December 2009

Target Organs: Reproduction

Thalidomide (Contergan)R

- late 50ies 1961/ 62 new drug for sleep disorders, tranquilizer - large step forward in term of low toxicity and almost vanished risks compared with established drugs (barbiturates) Wiedemann (1961): ununusal frequency of cases with amelia, phokomelia 4000 cases in germany / 7000 globally within few years the reason was clarified 13 mg/ kg body weight is effective

% &

7w

mating

birth treatment

end of lactation

2w Sectio ? Malformation, Postnatal Development Resorption Still birth fertility Multigeneration Test if needed
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2009-11-30

Needs for predictive Testing

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Heidi Foth, December 2009

Cancer Formation (1)

Human carcinogens polycyclic aromatic hydrocarbons aflatoxin B1 dimethyl nitrosamine 2-naphthylamine ethyl carbamate nickel carbonyl benzene vinylchloride

2009-11-30

Needs for predictive Testing

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Heidi Foth, December 2009

Cancer Formation (2)

Exposure Metabolic activation CYP450 induction adducts, oxidation, strand breaks, chromosomal aberration

Effects on gene expression DNA damage

repair, stress response altered gene expression

no effect visible Mutations in tumor genes

promotion, progression no effect visible CANCER 2009-11-30

Epigenetic control of cell cycle , apoptosis tumor suppressor genes

Needs for predictive Testing

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Heidi Foth, December 2009

(Proposed?) Strengths of in vitro based Assay

Compared to Animal based Standard test

Lower number of animals needed Save time Reproducibility of data Important to understand mechanisms of action Depict early signs of toxicity ( ? Search for biomarkers of effect?) Lower costs
2009-11-30 Alternative Methods 25

Heidi Foth, December 2009

In vitro Test battery covers major toxicity pathways

Cytotoxicity Oxidative/Cellular Stress Genotoxic Stress CYP/DME inductors Apoptosis inhibition Stress kinases/ Tumor promotion

various endpoints GSH/GSSG ratio GST activity p53 induction PXR-Luc (specific human CYP3A4 induction) CYP gene expression in primary hepatocytes (bDNA) ApoOne (caspase 3/7 activity) AP-1-Luc/NF-kB-Luc gene expression (cyclin D1, c-myc, GST-p and others) Source of information: Dr. Peter Jrgen Kramer, Merck

2009-11-30

Alternative Methods

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Heidi Foth, December 2009

Accepted in vitro-Methods
Endpoint Dermal Absorption Phototoxicity Irritation and Corrosion OECD Test Guideline Skin Absorption (428) 3T3 NRU Phototoxicity Test (432) Transcutaneous Electrical Resistance Test (430) Human Skin Model test (431) Membrane Barrier Test Method for Skin Corrosion (435) Embryonated chicken egg (HET-CAM) Isolated bovine cornea (BCOP) Isolated chicken eye (CEET) Isolated rabbit eye (IRE) Whole embryo culture (WEC) Micromass Test (MM) Embryonic stem cell test (EST)
Alternative Methods 27

Annex V Dir 67/548

Eye irritation

Embryotoxicity (prenatal development)

2009-11-30

Heidi Foth, December 2009

Overview on methods, accepted for regulatory purposes

Toxicological endpoint (No. der OECD Test Guideline) _________________________________________________________________________ Genotoxicity/ mutagenicity

In vivo
Mammalian erythrocyte micronucleus test 474 Mammalian bone marrow Chromosmal aberration test 475 Sex-linked recessive lethal test in drosophila 477 Rodent dominant lethal test 478 Mammalian spermatogonial Chromosomal aberration test 483 Mouse spot test 484 Mouse heritable translocation assay 485 Unscheduled DNA synthesis (UDS) test (mammalian liver) 486

In vitro
Bacterial reverse mutation test 471 Saccharomyces cervisiae Gene mutation assay 480 Mammalian chromosme aberration test 476 Mammalian cell gene mutation test 478 Micronucleus test 487 Mouse spot test 484 Sister chromatid exchange assay in mammalian cells 479 DNA damage and repair (UDS) in mammalian cells 482
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2009-11-30 CEC4, Rhodes, 2008

Alternative Methods Innovations & Challenges

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Heidi Foth, December 2009

Overview on methods, accepted for regulatory purposes

Toxicological endpoint (No. der OECD Test Guideline) _________________________________________________________________________ Carcinogenicity

In vivo
Carcinogenicity studies 451 Combined chronic toxicity Carcinogenicity Studies 453

In vitro
Cell transformation assay (SHE, Balb/c 3T3, C3H10T Test listed only in Annex V of Directive 67/548/EEC or accepted by Regulatory authorities of some EU member states

Full Carcinogenicity Studies Aim: Tumor formation after long treatment + genotoxicity - genotoxicity 500 animals / substance 1 000 000 and more limits sensitivity of species specificity of effect mechanism of effect duration up to 3.5 years
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2009-11-30 CEC4, Rhodes, 2008

Alternative Methods Innovations & Challenges

Heidi Foth, December 2009

Integrated Teststrategies unter REACH

Substance No exposure

Read-across

(Q)SARs

No further test

PBPK

in vivo-Tests

in vitro-Tests

Test Data needed for REACH Hartung et al. 2003

2009-11-30 Alternative Methods 30

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Heidi Foth, December 2009

In silico-Methods
SAR Structure activity relationships
SAR expert systems yes / no type of information QSAR statistical correlation
0 Predicted LogLC50 (mol/l) -1 -2 -3 -4 -5
P74 48 P N 21 N 16 N 39 P 64 N 24 N 30 N 17 P65 23N 31 N 56 P P 76 20 N 43 P 30 P N1 P 29 N N 33 66 N 54 N 32 P 85 N P 77 27 23 PN 78 42 PN 41 P 11 49 P 43 P 57 P 86 P N P42 1P 10 N 10 P 25 67 N 35 N 45 N 40 N 38 P P 79 P P 54 45 P19 46 P 31 P 32 P 33 N44 8 N P 28 P P 12 14 P N 4 36 P 58 P13 68 P 2 NN 15 P 26 P 53 P 24 61 P 25 N PP 9 59 63 22 N 57 PN 62 56 55 P P 15 P N8 29 P5 P 36 P 69 80 P81 51 P PP 27 N 28 P 35 P 37 P 47 N 6 N 37 P 70 P6 3P P 34 P 9 60 P 16 41 50 PP 39 P 18 N N 55 N 44 NP 14N P 17 P71 52 N PN 38 P26 40 P53 413 N 18 P P 20 P 722 N 12 46 P 50 19 72 N N 34 N N 47 N5 N 52 N 7 N2 N 11 N3 P 84 P 82 P 83 N 58 N 51 N 48 M O A - S et N PN PN

P 73 P 75 P 49 P 21

-6

There is hope, if

-7 -7 -6 -5 -4 -3 -2 -1 Expe rime nta l LogLC5 0 (mo l/ l) 0

The basic mechanism relies on easy to get physico-chemical properties of compounds The unknown substance belong to the group of training data set (domain of applicability) The endpoint of action is not complex The mechanisms of action is known ? The mechanisms of action are modelled by separate subsets ?
2009-11-30 Alternative Methods 31

Heidi Foth, December 2009

Application of QSAR in Regulation

Pharmaceuticals Screening and Prioritisation

Industrial Chemicals and others Behaviour and Fate in Environment

Intelligent Test Strategies if animal based testing is not applicable source of information for one end point

2009-11-30

Alternative Methods

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