Title of Research:16_S01-02
Prediction method (Amino acid Derivative Reactivity Assay: ADRA) for skin sensitization using novel
lysine and cysteine derivatives
Principal Investigator:
Toshihiko Kasahara, PhD (Engineering Manager, FUJIFILM Corporation, Safety Evaluation Center,
Ecology & Quality Management Division, CSR Division)
210, Nakanuma, Minamiashigara-shi, Kanagawa 250-0193, Japan(tel)+81-465-73-7440 (e-mail)ktoshihiko.kasahara@fujifilm.com
Collaborators:
Masaharu Fujita, PhD（FUJIFILM Corporation,Safety Evaluation Center）
210, Nakanuma, Minamiashigara-shi, Kanagawa 250-0193, Japan(tel)+81-465-73-7440 (e-mail)masaharu.fujita@fujifilm.com
Yusuke Yamamoto (FUJIFILM Corporation,Safety Evaluation Center）
210, Nakanuma, Minamiashigara-shi, Kanagawa 250-0193, Japan(tel)+81-465-73-7440 (e-mail)yusuke.yamamoto@fujifilm.com
Atsuishi Ono, PhD (Professor, Okayama University, Graduate school of Medicine, Dentistry andPharmaceutical sciences, Division of Pharmaceutical Sciences)1-1-1, Tsushimanaka, Kita-ku, Okayama-shi, Okanaya 700-8530, Japan(tel)086-251-7942 (e-mail) atsushiono3@okayama-u.ac.jp
Hajime Kojima, PhD (Director, National Institute of Health Sciences, Biological safety Research Center,
Division of Risk Assessment)
1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan(tel) 03-3700-9874 (e-mail)h-kojima@nihs.go.jp
Shinichi Wantanabe (LION CORPORATION, HUMAN & ENVIRONMENTAL SAFETY EVALUATIONCENTER)
100, Tajima, Odawara-shi, Kanagawa 256-0811(tel)0465-49-4475 (e-mail)shin-w@lion.co.jp
Tsunetsugu Sugawara (LION CORPORATION, HUMAN & ENVIRONMENTAL SAFETY EVALUATIONCENTER)
100, Tajima, Odawara-shi, Kanagawa 256-0811(tel)0465-49-4475 (e-mail) tsune-s@lion.co.jp
Koji Wakabayashi (Manager, MITSUI CHEMICALS, INC., Chemical Safety Department
1144, Togo, Mobara-shi, Chiba 297-0017, Japan(tel)0475-23-8251 (e-mail)Koji.Wakabayashi@mitsuichemicals.com
Yu Tahara (MITSUI CHEMICALS, INC., Chemical Safety Department
1144, Togo, Mobara-shi, Chiba 297-0017, Japan(tel)0475-23-8251 (e-mail)Yu.Tahara@mitsuichemicals.com
Nobuyuki Horie (Senior Research Scientist, SUMITOMO Chemical Co., Ltd., Environmental HealthScience Laboratory)
3-1-98, Kasugadenaka, Konohana-ku, Osaka-shi, Osaka 554-8558, Japan(tel)06-6466-5107 (e-mail)horien@sc.sumitomo-chem.co.jp
Keiichi Fujimoto (SUMITOMO CHEMICAL Co., Ltd., Environmental Health Science Laboratory)
3-1-98, Kasugadenaka, Konohana-ku, Osaka-shi, Osaka 554-8558, Japan(tel)06-6439-9753 (e-mail) fujimotok2@sc.sumitomo-chem.co.jp
Kei Kusakari (NISSAN CHEMICAL INDUSTRIES, LTD. Biological Research Laboratories)
1470, Shiraoka, Shiraoka-shi, Saitama 349-0294, Japan(tel)0480-92-2513 (e-mail)kusakarik@nissanchem.co.jp
Yoshihiko Kurokawa (NISSAN CHEMICAL INDUSTRIES, LTD. Biological Research Laboratories)
1470, Shiraoka, Shiraoka-shi, Saitama 349-0294, Japan(tel)0480-92-2513 (e-mail)kurokawayo@nissanchem.co.jp

Takashi Sozu, PhD (Assistant Professor, Department of Information and Computer Technology, Facultyof Engineering, Tokyo University of Science)
6-3-1, Shinjuku, Katsushika-ku, Tokyo 125-8585, Japan(tel)03-5876-1688 (e-mail)sozu@rs.tus.ac.jp
Takuto Nakayama (Department of Management Science, Graduate School of Engineering, Tokyo
University of Science)
6-3-1, Shinjuku, Katsushika-ku, Tokyo 125-8585, Japan(tel)03-5876-1688 (e-mail)4416624@ed.tus.ac.jp
Takeru Kusao (Department of Management Science, Faculty of Engineering, Tokyo University of Science)
6-3-1, Shinjuku, Katsushika-ku, Tokyo 125-8585, Japan(tel)03-5876-1688 (e-mail)j4413022@ed.tus.ac.jp
Tsuyoshi Kawakami, PhD (Director, National Institute of Health Sciences, Biological safety ResearchCenter, Division of Risk Assessment)
1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan(tel) 03-3700-1141 (e-mail)tkawa@nihs.go.jp
Kohichi Kojima、PhD （Director-General, Food and Drug Safety Center, Hatano Research Institute）
729-5, Ochiai, Hadano-shi, Kanagawa 257-8523, Japan(tel)0463-82-4751 (e-mail)kojima.k@fdsc.or.jp
Nicol C. Kleinstreuer, PhD （NTP Interagency Center for the Evaluation of Alternative Toxicologicalmethods）(tel) (919) 541-7997 (e-mail) nicole.kleinstreuer@nih.gov
Jon Richmind, PhD （Ethical Biomedical Research and Testing Advice & Consultancy）
Doubledykes Carslogie Road, Cupar, Fife KY154HY, United Kingdom(tel)077-86-381-757 (e-mail)dr.richmond@gmail.com
Bae-Hwan Kim, PhD （Professor, Keimyung University, Department of Public Health, College of NaturalScience）
1095, Dalgubeol-daero, Dalseo-Gu, Daegu 42601, Republic of Korea(tel)+82-53-580-5933 (e-mail)kim9399@kmu.ac.kr
Summary of Research:
In this study, we performed the validation study so that the alternative method for skin sensitization(ADRA) is adopted as OECD test guideline.
For the four participating laboratories, the two training sessions were held, and the two confirmation
tests (Pre-training test and Training test) were performed because the lead laboratory confirmed thatthe assay technique was correctly transferred for the participating laboratories.
The Pre-training test was performed using 5 test chemicals to be easy to be categorized correctly. The
predictive results for 5 chemicals at four participating laboratories were completely consistent with theresults at lead laboratory, so it was thought that the first step of technical transfer was finished. The
Training test was performed using 10 test chemicals that would be difficult to be categorized correctly.
The Within-Laboratory Reproducibility at each laboratory were 100%, 90%, 100% and 100%, respectively,
and so it was thought that technical transfer was completely finished because the goal wasaccomplished.
It was decided at VMT meeting that Phase-1 study (as Within-Laboratory Reproducibility) would beperformed using three replicate sets of 10 chemicals and Phase-2 (as Between-Laboratory
Reproducibility) study would be performed using one set of 30 chemicals in the ADRA validation study.
The Within-Laboratory Reproducibility were 100% (10/10) , 100%(10/10), 100%(7/7) and 90%(9/10) andthese values were higher than expected in that the results of Phase-1 achieved the Within-Laboratory
Reproducibility of more than 80% of target.
After Phase-1 validation study, VMT teleconference was held on January in 2019, and VMT member
were evaluated the results of Phase-1 study. Because all the improvement items proposed by leadlaboratory were approved by VMT members, and the Phase-2 study was started on February in 2019 asscheduled.

Timeline:March 1, 2016Topics:
1. Presented at ICCA-LRI & NIHS WORKSHOP-Awaji Island, Japan 「Validation study on Amino acid
Derivative Reactivity Assay (ADRA) for prediction of skin sensitization」
Publications:
1. Presented at the 29th Annual Meeting of the Japanese Society of Alternatives to AnimalExperiments (November 18, 2016) 「Reporting of Training and Transfer Outcome for Validation
Study of Alternative Method (ADRA) for Skin Sensitization using Cys and Lys Derivatives」
2. Planed to be presented at the 44th Annual Meeting of the Japanese Society of Toxicology(July 10-12, 2017) 「Multicenter Validation Study of Novel in chemico Skin Sensitization
Assay (ADRA): 1st Report」