Guidance on use of alternative methods for testing in the safety assessment of

cosmetics and quasi-drug

Hajime Kojima, NIHS, et al.

1

Asian Congress 2016, Nov. 15, 2016, Karatsu, Saga, Japan

2

JCIA safety evaluation

guidance

（2015）

PCPC safety evaluation guideline

（2014）

Cosmetic Europe, COLIPA

guideline（2004）

Single dose toxicity Oral toxicity Acute toxicity (oral or inhalation)

Repeat dose toxicity Dermal toxicity Sub-chronic toxicity (oral or inhalation)

－ Inhalation toxicity

Primary skin irritation Primary dermal irritation

Dermal irritation

Cumulative skin irritation － －

Skin sensitization Dermal sensitization Skin sensitization

Phtotoxicity Phtotoxicity& Photoallegy Phtotoxicity

Photosensitization

Ocular irritation Eye irritation Eye irritation

Genotoxicity Genotoxicity Mutagenicity

Human patch test Controlled use studies in human

Human data

－ － Toxicokinetics

－ Mucous membrane irritation Mucous membrane irritation

Skin absorption

Skin absorption Dermal absorption

Reproductive and developmental

toxicity

Reproductive and developmental toxicity

Reproductive toxicity, Carcinogenecity,

Additional genotoxicity

Table 1. Test methods for safety evaluation of cosmetic ingredients defined by each cosmetic industry association.

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Quasi-drug safety

evaluation guidance

（2008）

SCCS safety

evaluation guidance

（2012）

Special cosmetic

safety evaluation

guideline

Functional cosmetic

safety evaluation

guideline

Acute toxicity Acute toxicity Acute toxicity

Repeat dose toxicity (Sub-

chronic, Chronic)

Repeat dose toxicity

Reproductive toxicity Reproductive toxicity

Skin irritation Corrosivity & irritation Acute skin irritation Primary skin irritation

－ Cumulative skin irritation －

Skin sensitization Skin sensitization Skin sensitization Skin sensitization

Phtoto-induced toxicity Phtotoxicity

Photo sensitization Photo sensitization Photo sensitization

Mucous membrane irritation

Acute eye irritation Eye or mucous membrane

irritation

Genotoxicity Mutagenicity/genotoxicity Genotoxicity

Human data Human patch test Human patch test

Controlled use studies in

human

Human repeat insult patch

test

ADME Toxicokinetics

Dermal/percutaneous

absorption

Carcinogenecity Carcinogenecity

Table 2. Test methods for safety evaluation of ingredients for regulatory use

Regulatory issue on alternative methods

• According to the MHLW notification, Japan in 2011, JaCVAM (Japanese Center for the Validation of Alternative Methods) decided to accelerate new in vitro testing methods to take advantage of this opportunity to strongly impact testing throughout Japan.

• Therefore, the members are coordinating the Guidance on the use of alternative test methods in safety assessment of cosmetics and quasi-drugs since 2012.

• The members have consists on dermatologists, delegates of cosmetic companies, the technical officers of PMDA (Pharmaceuticals and Medical Devices Agency) and specialists of NIHS (National Institute of Health Sciences) and this member is on-going to make drafting the guidance based on the OECD test guideline and/or JaCVAM evaluation document per each alternative test method.

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Members for the guidance

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Classification Name Affiliation

Dermatologist Tokio Nakada Showa University, Fujigaoka Hospital, Japan

Akiko Yagami Fujita Health University School of Medicine, Japan

Regulator Yukiko Hoshino Pharmaceuticals and Medical Devices Agency, Japan Naofumi Iizuka

Takatoshi Nakamura

Shinichi Sekizawa

Kazutoshi Shinoda

Mio Yagi

Researcher Kenji Sugibayashi Faculty of Pharmaceutical Sciences, Josai University, Japan Hiroaki Todo

Yoshiaki Ikarashi National Institute of Health Sciences, Japan Hajime Kojima

Industry Daisuke Araki Japan Cosmetic Industry Association, Japan Hitoshi Sakaguchi

Hitoshi Sasa

Mariko Sugiyama

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On-going draft guidance

• Complete of public comment on in vitro skin absorption

• Approach to a guidance on non-animal testings for skin sensitization

• Start discussion for Short Time Exposure assay for alternative to eye irritation testing

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Approach to a guidance on non-animal testings for skin sensitization

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ENV/JM/HA(2016)11 – Annex 1

Case study Purpose 1 An Adverse Outcome Pathway-based "2 out of 3" integrated testing

strategy approach to skin hazard identification (BASF) Hazard

identification

2 Sequential Testing Strategy (STS) for hazard identification of skin sensitisers (RIVM)

Hazard identification

3 A non-testing Pipeline approach for skin sensitisation (G. Patlewicz)

Hazard identification

4 Stacking meta-model for skin sensitisation hazard identification (L'Oréal) Hazard identification

5 Integrated decision strategy for skin sensitisation hazard (ICCVAM)

Hazard identification

6 Consensus of classification trees for skin sensitisation hazard prediction (EC- JRC)

Hazard identification

7 Sensitizer potency prediction based on Key event 1 + 2: Combination of kinetic peptide reactivity data and KeratinoSens® data (Givaudan)

Potency prediction

8 The artificial neural network model for predicting LLNA EC3 (Shiseido) Potency prediction

9 Bayesian Network DIP (BN-ITS-3) for hazard and potency identification of skin sensitizers (P&G)

Potency prediction

10 Sequential testing strategy (STS) for sensitising potency classification based on in chemico and in vitro data (Kao Corporation)

Potency prediction

11 Integrated testing strategy (ITS) for sensitising potency classification based on in silico, in chemico, and in vitro data (Kao Corporation)

Potency prediction

12 DIP for skin allergy risk assessment (SARA) (Unilever)

Potency prediction 10

Artificial Neural Network (ANN) analysis

Input layer Output layer

. . .

Hidden layer

LLNA EC3

In vitro test data

In vivo test data

Other biomarkers

h-CLAT

DPRA-Cys

DPRA-Lys

KeratinoSens

• EC3 values for LLNA

positive chemicals

• For LLNA negative

chemicals, 101% was

used as EC3 value

(converted to mM/cm2 unit).

dendrite

axon synapses

・EC3 values for LLNA positive chemicals ・For LLNA negative chemicals, 101% was used as EC3 value

(converted to mM/cm2 unit).

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This ANN model showed a good correlation between predicted values and actual values. Therefore, our ANN model can contribute to building a new QRA evaluation system which is using no animals.

*Root mean square (RMS) error = data ofnumber / ) value)predicted - value((measured 2

- R= 0.87(N=134)

RMS error=0.57

Log(Published LLNA EC3 (uM/cm2))

-

- -

Log(P

redic

ted L

LN

A E

C3 (

uM

/cm

2)

)

Correlation between LLNA and the ANN prediction

After 10-fold

cross-validation

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Kao Sequential (Tiered) Testing Strategy (STS)

Kao Integrated (Score-based) Testing Strategy (ITS)

Takenouchi et al. 2015

DPRA (KE1) and h-CLAT (KE3) Predicting the three potency classes

not classified, weak (EC3 ≥ 1%), strong (EC3 < 1%)

DPRA, Derek (KE1) and h-CLAT (KE3) Predicting the three potency classes

not classified, weak (EC3 ≥ 1%), strong (EC3 < 1%)

Total score of 0-1: not classified, of 2-6: weak, of 7: strong

Kao STS & ITS

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Kao STS (n=139)

Kao ITS (n=139)

Hazard accuracy：81% (10 FN, 17 FP) Potency accuracy : 69% (20 under, 23 over)

Hazard accuracy：84% (11 FN, 11 FP) Potency accuracy : 71% (18 under, 12 over)

Kao STS & ITS compared to LLNA data

Kao STS (n=129)

Kao ITS (n=128)

Hazard accuracy：85% (2 FN, 17 FP) Potency accuracy : 73%

Hazard accuracy：89% (3 FN, 11 FP) Potency accuracy : 74%

Takenouchi et al. 2015

Excluding negative results of chemicals with logKow>3.5

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3 out of 3 for Bottom-up

DPRA

Negative

KeratinoSens

Negative

h-CLAT

Negative

LLNA 91 64 82 - - - 111

3 out of 3 for

bottom-up97 35 81 99 43 86 100

Sensitivity

(%)

vs LLNA

Sensitivity

(%)

Specificity

(%)

Accuracy

(%)

vs human data

NSpecificity

(%)

Accuracy

(%)

Non-sensitizer

Analyzed using data from Urbisch et al. 2015

Inconsistent results of in vitro assays would require additional information to support the conclusion.

“3 out of 3 for Bottom-up” is not the only approach. Additional testing strategies to identify non-sensitizers are also under discussion.

“3 out of 3 for Bottom-up” could be used as “one” of testing strategies to identify non-sensitizers as part of bottom-up approach

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Summary

The members of guidance team have also contributed to the establishment of a guidance for skin sensitization based on Test Guidelines (TGs) and guidance documents in OECD.

Thank you for your attention

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