ホ౯఍㆟ሗ࿌᭩⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࣄࢺ⾲⓶ࣔࢹࣝἲ

JaCVAM ホ౯఍㆟

ᖹᡂ 29 ᖺ㸦2017 ᖺ㸧5 ᭶ 11 ᪥11JaCVAM ホ౯఍㆟኱㔝Ὀ㞝㸦බ┈㈈ᅋἲே ᮌཎグᛕᶓ὾⏕࿨⛉Ꮫ᣺⯆㈈ᅋ㸧㸸ᗙ㛗஬༑ᔒⰋ᫂㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ ⏕ά⾨⏕໬Ꮫ㒊㸧▼ ஭ 㞝 ஧ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧஭ୖᬛᙲ㸦᪥ᮏච␿ẘᛶᏛ఍㸧௒஭ᩍᏳ㸦᪥ᮏື≀ᐇ㦂௦᭰ἲᏛ఍㸧ᒾ℩⿱⨾Ꮚ㸦᪥ᮏ〇⸆ᕤᴗ༠఍㸧⠛⏣࿴ಇ㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧ᮡᒣ┿⌮Ꮚ㸦᪥ᮏ໬⢝ရᕤᴗ㐃ྜ఍㸧௰஭ಇྖ㸦᪥ᮏ໬Ꮫᕤᴗ༠఍㸧୰ᮧࡿࡾࡇ

㸦⊂❧⾜ᨻἲே 〇ရホ౯ᢏ⾡ᇶ┙ᶵᵓ㸧㔝 ཱྀ ┿ ᕼ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧すᕝ⛅ె㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧἟⃝ ⪽㸦᪥ᮏẘᛶᏛ఍㸧᳃⏣ ೺㸦᪥ᮏ⎔ቃኚ␗ཎᏛ఍㸧ᶓ㛵༤㞝㸦᪥ᮏ⓶⭵࢔ࣞࣝࢠ࣮࣭᥋ゐ⓶⭵⅖Ꮫ఍㸧                ௵ᮇ㸸ᖹᡂ 28 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 30 ᖺ 3 ᭶ 31 ᪥22ࣄࢺ⾲⓶ࣔࢹࣝࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂ἲࡣࠊ࢘ࢧࢠࢆ⏝࠸ࡿ⓶⭵⭉㣗ᛶヨ㦂ࡢ௦᭰ἲ࡜ࡋ࡚㛤Ⓨࡉࢀࡓヨ㦂ἲ࡛࠶ࡿࠋᮏヨ㦂ἲ࡛ࡣࠊ⭉㣗ᛶ≀㉁ࡀゅ㉁ᒙࢆയᐖࡲࡓࡣゅ㉁ᒙ࡟྾཰ࡉࢀࡓᚋᣑᩓࡋࠊ⾲⓶⣽⬊࡟฿㐩ࡋ࡚⣽⬊ẘᛶࢆ♧ࡍ࡜࠸࠺௬ㄝ࡟ᇶ࡙ࡁࠊ⿕㦂≀㉁᭚㟢ᚋࡢ⣽⬊⏕Ꮡ⋡ࢆᣦᶆ࡟⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿࠋᮏヨ㦂ἲ࡟ࡘ࠸࡚ࡣࠊEpiSkin™ࠊEpiDerm™ࠊSkinEthic™ࠊepiCS®࡜࠸࠺㸲✀ࡢ⾲⓶ࣔࢹࣝࡀ࠶ࡆࡽ
ࢀ࡚࠸ࡿࠋ࠸ࡎࢀࡢࣔࢹࣝ࡟ࡼࡿヨ㦂ࡶࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࠊECVAM㸦European Centre forthe Validation of Alternative Methods㸸Ḣᕞ௦᭰ἲホ౯ࢭࣥࢱ࣮㸧࡟ࡼࡾࡑࡢಙ㢗ᛶ࡜෌⌧ᛶࡀ㧗࠸ࡇ࡜ࡀ☜ㄆࡉࢀࡓ 1~4)ࠋEpiSkin™ࠊEpiDerm™࡟࠾࠸࡚ࡣࠊESAC㸦ECVAM Scientific Advisory Committee㸸ECVAM⛉Ꮫㅎၥ఍㆟㸧࡛ࡢホ౯ᚋࠊࡑࡢ⤖ㄽࡣ ICCVAM㸦Interagency Coordinating Committee on the Validation ofAlternative Methods㸸⡿ᅜ௦᭰ἲ࡟㛵ࡍࡿ┬ᗇ㛫㐃⤡఍㆟㸧࡟࠾࠸࡚ࡶ☜ㄆࡉࢀࡓ 5,6) ࠋୖグ 4 ✀ࣔࢹࣝ࡟ࡘ࠸࡚ࡣ OECD㸦Organisation for Economic Co-operation and Development㸸⤒῭༠ຊ㛤Ⓨᶵᵓ㸧࡟࡚ࢸࢫࢺ࢞࢖ࢻࣛ࢖ࣥ㸦TG㸧431 ࡜ࡋ࡚ᢎㄆࡉࢀࡓࠋࡇࡢ TG ࡣ᫖௒✀ࠎࡢⅬ࡛ᨵゞࡉࢀ࡚࠾ࡾࠊ⌧ᅾࡣ 2016ᖺ∧࡜࡞ࡗ࡚࠸ࡿ 7)ࠋ
JaCVAM ホ౯఍㆟ࡣࠊ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍࡟ࡼࡾసᡂࡉࢀࡓࠕࣄࢺ⾲⓶ࣔࢹࣝࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢホ౯ሗ࿌᭩ࠖ8)ࢆ⏝࠸࡚ࠊᮏヨ㦂ἲࡢጇᙜᛶ࡟ࡘ࠸᳨࡚ウࡋࡓࠋ㸯㸬ヨ㦂ἲࡢᐃ⩏ྡ⛠㸸 ࣄࢺ⾲⓶ࣔࢹࣝࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ௦᭰ࡍࡿᑐ㇟ẘᛶヨ㦂㸸 ࢘ࢧࢠࢆ⏝࠸ࡿ⓶⭵⭉㣗ᛶヨ㦂ヨ㦂ἲࡢᴫ␎㸸ᮏヨ㦂ἲ࡛ࡣࠊ࢘ࢧࢠ⓶⭵ࡢ௦ࢃࡾ࡟ゅ㉁ᒙࢆ᭷ࡍࡿ 3 ḟඖ෌ᵓ⠏ࣄࢺ⾲⓶ࣔࢹࣝࢆ⏝࠸ࠊ⿕㦂≀㉁ࡀゅ㉁ᒙୗࡢ⾲⓶⣽⬊ᒙ࡟ᑐࡋ⣽⬊ẘᛶࢆ♧ࡍ⬟ຊࢆホ౯ࡍࡿࠋ⾲⓶ࣔࢹࣝ⾲㠃࡟⿕㦂≀㉁ࢆ୍ᐃ᫬㛫㐺⏝ࡋࡓᚋὙίࡋࠊ⾲⓶⣽⬊ࡢ⏕Ꮡ⋡ࢆ MTTࠝ3 ( 4, 5 - Dimethylthiazol - 2 - yl ) - 2, 5-diphenyltetrazolium bromideࠞࡢ㑏ඖ㔞࠿ࡽồࡵࠊ⓶⭵⭉㣗ᛶࢆุᐃࡍࡿࠋ㸰㸬ホ౯࡟⏝࠸ࡓ㈨ᩱཬࡧホ౯ෆᐜࡢ⛉Ꮫⓗጇᙜᛶᮏヨ㦂ἲࡣࠊ4 ✀ࡢ⾲⓶ࣔࢹࣝࡢ࠸ࡎࢀࡢࣔࢹࣝࡶࠊECVAM ࡟ࡼࡿࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡜ࡑࢀ࡟⥆ࡃ ESAC ࡟ࡼࡿ➨୕⪅ホ౯࡟ࡼࡾࠊᐇ㦂ື≀ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂ࡢ௦᭰ἲ࡜ࡋ࡚⛉Ꮫⓗ࡟ጇᙜ࡛࠶ࡿ࡜ሗ࿌ࡉࢀ࡚࠾ࡾ 1~4)ࠊEpiSkin™࠾ࡼࡧ EpiDerm™ࢆ⏝࠸ࡓヨ㦂ἲ࡟ࡘ࠸࡚ࡣ ICCVAM ࡟࠾࠸࡚ࡶྠᵝ࡟☜ㄆࡉࢀࡓ 6)ࠋୖグ 4 ✀ࡢ⾲⓶ࣔࢹࣝࢆ⏝࠸ࡿ᪉ἲ࡟ࡘ࠸࡚ࡣ⌧ᅾ OECD ࢸࢫࢺ࢞࢖ࢻࣛ࢖ࣥ࡜ࡋ࡚ᢎㄆࡉࢀ࡚࠸ࡿ 7)ࠋJaCVAM ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍࡛ࡣࠊ⌧ᅾࡲ࡛බ㛤ࡉࢀ࡚࠸ࡿ᝟ሗ 1~7,9~11)ࢆᇶ࡟ࣄࢺ⾲⓶ࣔࢹࣝࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ࡜ࡋ࡚ࡢ⛉Ꮫⓗጇᙜᛶ࡟ࡘ࠸࡚ホ౯ࡋࡓࠋࡑࡢ⤖ᯝࠊᮏヨ㦂ἲࡣࠊࠕ⭉㣗ᛶ≀㉁ࡀゅ㉁ᒙࡢയᐖࡲࡓࡣゅ㉁ᒙ࡬ࡢ྾཰ࢆ⤒࡚ୗᒙࡢ⾲⓶⣽⬊࡟ᑐࡋẘᛶࢆ♧ࡍࠖ࡜࠸࠺⓶⭵⭉㣗ᛶⓎ⌧ᶵᗎ࡟ᇶ࡙ࡁࠊ⣽⬊࡬ࡢẘᛶࢆᣦᶆ࡟ࡋࡓࡶࡢ࡛࠶ࡾࠊཎ⌮ⓗ࡟ࡶጇᙜ࡛࠶ࡿ࡜ุ᩿ࡉࢀࡓࠋ

33㸱㸬ᮏヨ㦂ἲࡢ᭷⏝ᛶ࡜㐺⏝㝈⏺
࠸ࡎࢀࡢ⾲⓶ࣔࢹࣝࡶື≀ࢆ౑⏝ࡋ࡚࠾ࡽࡎࠊື≀⚟♴㠃࠿ࡽ௦᭰ἲ࡜ࡋ࡚ጇᙜ࡛࠶ࡿࠋᙉ࠸㓟ᛶ㸦pH2.0 ௨ୗ㸧ࡲࡓࡣ࢔ࣝ࢝ࣜᛶ㸦pH 11.5 ௨ୖ㸧≀㉁ࡣࠊᙉ࠸ᒁᡤയᐖࢆᘬࡁ㉳ࡇࡍྍ⬟ᛶࡀ࠶ࡿࡇ࡜࠿ࡽࠊ⓶⭵⭉㣗ᛶ࡜ุ᩿ࡋ࡚ࡶⰋ࠸ࡇ࡜࡟࡞ࡗ࡚࠸ࡿ 9)ࠋࡋ࠿ࡋ࡞ࡀࡽࠊࡇࢀࡣ⭉㣗ᛶ࡟ࡘ࠸࡚ࡢ᝟ሗࡀ௚࡟࡞࠸ሙྜ࡟⾜ࢃࢀࡿ࣮࣡ࢫࢺࢣ࣮ࢫ࡜ࡋ࡚ࡢุ᩿࡛࠶ࡾࠊ౛࠼ࡤ⦆⾪స⏝ࡀᑠࡉ࠸≀㉁ࡸΰྜ≀ࡢሙྜࡣഇ㝧ᛶࡢุ᩿࡜࡞ࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿ 10)ࠋࡇࡢࡓࡵࠊࡇࡢࡼ࠺࡞≀㉁࡟ᑐࡋ࡚ࡶື≀ࢆ⏝࠸ࡿᚲせࡢ࡞࠸ᮏヨ㦂ἲ࡛⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿࡇ࡜ࡣ᭷⏝࡛࠶ࡿࠋ࠸ࡎࢀࡢࣔࢹࣝࡶ UN GHS (United Nations Globally Harmonized System of Classification and Labelling ofChemicals㸸ᅜ㐃 GHS) ศ㢮࡟࠾ࡅࡿ⭉㣗ᛶ㸦༊ศ 1㸧࡟ࡘ࠸࡚ࠊࡑࡢ᭷↓ࡢணᛶࡣ㧗ࡃࠊOECD ࡛㞟ィࡋࡓ໬Ꮫ≀㉁࡛ࡣࠊឤᗘࡣ 95㹼100㸣ࠊ≉␗ᗘࡣ 72㹼79㸣ࠊṇ☜ᗘࡣ 84㹼90%࡛࠶ࡾࠊഇ㝜ᛶ⋡ࡣ 0
㹼5%࡜ప࠸࡛ࣞ࣋ࣝ࠶ࡗࡓ㸦⾲ 1㸧ࠋྛࣔࢹࣝ࡟ࡼࡿᅜ㐃 GHS ศ㢮ࡢ⣽༊ศ㸦1A㹼1C㸧ࡢணᛶ࡟ࡘ࠸࡚ࡣࠊ80 ≀㉁ࢆ 2 ࡲࡓࡣ 3 ᅇᐇ㦂ࡋ࡚ᚓࡽࢀࡓ್ࢆぢࡿ㝈ࡾࠊEpiSkinTM ࡢࡳࡀ 1B/1C ࢆ 80㸣⛬ᗘ㆑ูྍ⬟࡛࠶ࡗࡓࠋࡑࡢ௚ࡢࣔࢹࣝࡣ⣽༊ศࡢホ౯࡟ࡣ฼⏝࡛ࡁ࡞࠸࡜ุ᩿ࡋࡓࠋ
MTT 㑏ඖ≀㉁࡟ᑐࡋ࡚ࡣࠊTG431 ࡟ᑐᛂ᪉ἲࡀグ㍕ࡉࢀ࡚࠾ࡾࠊࡇࢀ࡟‽ࡌࡿࡇ࡜࡛ホ౯ྍ⬟࡛࠶ࡿࠋ࢞ࢫࠊ࢚࢔ࣟࢰ࣮ࣝ࡟ࡘ࠸࡚ࡣࣂࣜࢹ࣮ࢩࣙࣥࡀᐇ᪋ࡉࢀ࡚࠾ࡽࡎࠊ㐺⏝ྍྰࡣุ᩿࡛ࡁ࡞࠸ࠋࡑࢀࡽ௨እࡢ≀㉁࡟ࡘ࠸࡚ࡣࠊ≀⌮≧ែ㸦ᾮయ࣭ᅛయ➼㸧࠾ࡼࡧỈ⁐ᛶࡢ᭷↓࡟࠿࠿ࢃࡽࡎ㐺⏝ྍ⬟࡛࠶ࡾࠊΰྜ≀࡟ࡶ㐺⏝ྍ⬟࡞ሙྜࡀ࠶ࡿࠋ⾲ 1㸬໬Ꮫ≀㉁ࡢ඲ࢭࢵࢺ*࡟࠾ࡅࡿணᛶ㸦⭉㣗ᛶ [ᅜ㐃 GHS ༊ศ㸯] ࡢ᭷↓㸧11)

EpiSkinTMEpiDermTM

SkinEthicTM

epiCS®

ṇ☜ᗘ

89.6㸣

87.9%

84.6㸣

84.3㸣

ឤᗘ98.5㸣

100%

94.6㸣

95.3㸣

≉␗ᗘ

79.3㸣

73.9%

73.0㸣

71.6㸣

ഇ㝧ᛶ⋡

20.7%

26.1%

27.0%

28.4%

ഇ㝜ᛶ⋡

1.5%

0%

5.4%

4.7%

*) OECD ࡢ㞟ィ࡟ࡼࡿࠋ㸲㸬┠ⓗ࡜ࡍࡿ≀㉁ཪࡣ〇ရࡢẘᛶࢆホ౯ࡍࡿヨ㦂ἲ࡜ࡋ࡚ࡢࠊ♫఍ⓗཷࡅධࢀᛶཬࡧ⾜ᨻୖࡢ฼⏝ࡢྍ⬟ᛶ ♫఍ⓗཷࡅධࢀᛶ㸸ᮏヨ㦂ἲࡣࠊ㏻ᖖࡢᇵ㣴ᢏ⾡ࢆ⩦⇍ࡋࡓ᪋タ࡛࠶ࢀࡤᐇ᪋࡛ࡁࡿヨ㦂ἲ࡛࠶ࡾࠊOECD TG431 ࡟ᢎㄆࡉࢀࡓ⾲⓶ࣔࢹࣝࡣᕷ㈍ࡉࢀ࡚࠸ࡿࠋࡲࡓࠊ⏕ࡁࡓື≀ࢆ⏝࠸࡞࠸࡜࠸࠺Ⅼ࡛ࠊ3Rs ࡢ⢭⚄࡟ྜ⮴ࡋ࡚࠾ࡾࠊ♫఍ⓗཷࡅධࢀᛶࡣ㧗࠸ࠋ⾜ᨻୖࡢ฼⏝ᛶ㸸ᮏヨ㦂ἲ࡟࠾࠸࡚㝧ᛶࡢ⤖ᯝࡀᚓࡽࢀࡓሙྜࠊ⿕㦂≀㉁ࢆ⭉㣗ᛶ≀㉁㸦ᅜ㐃 GHS ศ㢮࡟࠾ࡅࡿ༊ศ1㸧࡜ุᐃࡍࡿࡇ࡜ࡣྍ⬟࡛࠶ࡿࡀࠊUN GHS ศ㢮ࡢ⣽༊ศࡢࡓࡵ࡟ࡣ EpiSkinTM ࡢࡳ฼⏝ྍ⬟࡛࠶ࡿࠋ࡞࠾ࠊᮏヨ㦂ἲࡢ฼⏝࡟࠶ࡓࡗ࡚ࡣࠊ㐺⏝⠊ᅖࢆ༑ศ࡟㓄៖ࡋࡓୖ࡛౑⏝ࡉࢀࡿ࡭ࡁ࡛࠶ࡿࠋ

44ཧ⪃ᩥ⊩1)

ECVAM (1998) Statement on the scientific validity of the EpiSkin test (An in vitro test for skin corrosivity)

2)

ECVAM (2000) Statement on the application of the EpiDermTM human skin model for skin corrosivity testing

3)

ECVAM (2006) Statement on the application of the SkinEthicTM human skin model for skin corrosivity testing

4)

ECVAM (2009) ESAC Statement on the scientific validity of an in-vitro test for skin corrosivity testing

5)

ICCVAM (1999) NIH Publication No.99-4495. Corrositex: An in vitro test method for assessing dermalcorrosivity potential of chemicals.

6)

ICCVAM (2002) NIH Publication No.02-4502. ICCVAM Evaluation of EPISKINTM, and EpiDermTM (EPI200) and rat skin transcutaneous electrical resistance (TER) assay: in vitro test method for assessing dermal
corrosivity potential of chemicals.

7)

OECD (2016) Guideline for the testing of chemicals. 431, in vitro Skin Corrosion: Human skin model test.

8)

JaCVAM ⓶⭵⭉㣗ᛶ㈨ᩱ⦅⧩ጤဨ఍㸸⓶⭵⭉㣗ᛶヨ㦂ホ౯ሗ࿌᭩ ࣄࢺ⾲⓶ࣔࢹࣝࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢホ౯ሗ࿌᭩㸦2017 ᖺ 2 ᭶ 24 ᪥㸧㸬9)OECD (2014) Series on Testing and Assessment No. 203, Guidance document on an Integrated Approach onTesting and Assessment (IATA) for skin corrosion and irritation

10) Sheel J. et.al., Classification and labeling of industrial products with extreme pH by making use of in vitro
methods for the assessment of skin and eye irritation and corrosion in a weight of evidence approach,Toxicology in Vitro, 25, 1435-1447 (2011)11) OECD (2015) Series on Testing & Assessment No. 219, Performance Standards for the assessment of proposed
similar or modified in vitro reconstructed human epidermis (RhE) test methods for skin corrosion testing asdescribed in TG 43155