Title
カニクイザルでの網膜電図を用いた薬剤誘発性網膜機能障
害の評価に関する研究 ( 本文(Fulltext) )
Author(s)
木下, 順三
Report No.(Doctoral
Degree)
博士(獣医学) 乙第129号
Issue Date
2014-03-13
Type
博士論文
Version
ETD
URL
http://hdl.handle.net/20.500.12099/49050
※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。࢝ࢽࢡࢨ࡛ࣝࡢ⥙⭷㟁ᅗࢆ⏝࠸ࡓ⸆ㄏⓎᛶ⥙⭷ᶵ⬟㞀ᐖࡢホ౯
㛵ࡍࡿ◊✲
2013 ᖺ
ᒱ㜧ᏛᏛ㝔㐃ྜ⋇་Ꮫ◊✲⛉
┠
┠ḟ
␎ㄒ୍ぴ ...ii
⥴ゝ... 1
➨1 ❶ Standard full-field ERG㸸ᮞయ⤒㊰ཬࡧ㗹య⤒㊰ࡢศ㞳ホ౯... 9
1. ᗎㄽ ... 9 2. ᐇ㦂ᮦᩱཬࡧᐇ㦂᪉ἲ... 11 3. ᐇ㦂⤖ᯝ... 14 4. ⪃ᐹ ... 15 5. ᑠᣓ ... 17 ➨2 ❶ a-wave analysis㸸୍ḟࢽ࣮ࣗࣟࣥ㸦ど⣽⬊㸧ࡢホ౯ ... 22 1. ᗎㄽ ... 22 2. ᐇ㦂ᮦᩱཬࡧᐇ㦂᪉ἲ... 23 3. ᐇ㦂⤖ᯝ... 28 4. ⪃ᐹ ... 30 5. ᑠᣓ ... 33 ➨3 ❶ ON-OFF response㸸ḟࢽ࣮ࣗࣟࣥ㸦ᴟ⣽⬊㸧ࡢホ౯ ... 45 1. ᗎㄽ ... 45 2. ᐇ㦂ᮦᩱཬࡧᐇ㦂᪉ἲ... 46 3. ᐇ㦂⤖ᯝ... 49 4. ⪃ᐹ ... 51 5. ᑠᣓ ... 52
➨4 ❶ Photopic negative response㸸୕ḟࢽ࣮ࣗࣟࣥ㸦⥙⭷⚄⤒⠇⣽⬊㸧ࡢホ౯ ... 58
1. ᗎㄽ ... 58 2. ᐇ㦂ᮦᩱཬࡧᐇ㦂᪉ἲ... 59 3. ᐇ㦂⤖ᯝ... 62 4. ⪃ᐹ ... 64 5. ᑠᣓ ... 65 ⥲ᣓ... 76
ii
␎ㄒ୍ぴ
ᮏᩥཬࡧᅗ⾲࡛⏝࠸ࡓ␎ㄒࡣ௨ୗࡢ㏻ࡾ࡛࠶ࡿࠋAPB: 2-amino-4-phosphonobutyric acid
AUC0-inf: Area under the concentration-time curve up to infinity
AUCtau: Area under the concentration-time curve during dosing interval
AUCtau,ss: Area under the concentration-time curve during dosing interval at steady state
AUC0-24h: Area under the concentration-time curve up to 24 hours after dosing
cGMP: Cyclic guanosine monophosphate
Cmax: Maximum concentration
ERG: Electroretinogram
ICH: International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use
iGluR: Ionotropic glutamate receptor
ISCEV: International Society for Clinical Electrophysiology of Vision L-cones: Long wavelength sensitive cones
LED: Light-emitting diode
M-cones: Middle wavelength sensitive cones mGluR6: Metabotropic glutamate receptor type 6
ND: Neutral density
NMDA: N-methyl-D-aspartic acid
PDA: cis-2,3-piperidine dicarboxylic acid
PDE5: Phosphodiesterase type 5
PDE6: Phosphodiesterase type 6
PhNR: Photopic negative response
QOL: Quality of life
RGC: Retinal ganglion cell
S-cones: Short wavelength sensitive cones
TK: Toxicokinetics
⥴ゝ
⥙⭷ࡣ║⌫ቨࡢ᭱ࡶෆഃ⨨ࡍࡿ⭷⤌⧊࡛࠶ࡾ㸦Fig. 1㸧ࠊ⤌⧊Ꮫⓗࡣእഃࡼࡾࠊ⥙⭷ Ⰽ⣲ୖ⓶ࠊᮞయ㗹యᒙࠊእቃ⏺⭷ࠊእ㢛⢏ᒙࠊእ⥙≧ᒙࠊෆ㢛⢏ᒙࠊෆ⥙≧ᒙࠊ⚄⤒⠇⣽ ⬊ᒙࠊ⚄⤒⥺⥔ᒙࠊཬࡧෆቃ⏺⭷ࡢ10 ᒙࡽ࡞ࡿ㸦Fig. 2㸧ࠋࡇࡢ࠺ࡕ⥙⭷Ⰽ⣲ୖ⓶௨እࡢ 9 ᒙࡣឤぬ⥙⭷ࡤࢀࠊゅ⭷ࡸỈᬗయࡼࡗ࡚ᵓᡂࡉࢀࡿ║⌫ࡢගᏛ⣔ࡼࡗ࡚ᒅᢡࡋࡓ ീࡀࡇࡢឤぬ⥙⭷⤖ീࡍࡿࠋ⥙⭷㐩ࡋࡓගࡣࠊ୍ḟࢽ࣮࡛ࣗࣟࣥ࠶ࡿど⣽⬊ࡼࡗ࡚ ⭷㟁ࡢኚ࠸࠺㟁Ẽಙྕኚࡉࢀࡿࠋࡑࡢどぬሗࡣࡉࡽࠊḟࢽ࣮࡛ࣗࣟࣥ࠶ ࡿᴟ⣽⬊ཬࡧ୕ḟࢽ࣮࡛ࣗࣟࣥ࠶ࡿ⥙⭷⚄⤒⠇⣽⬊㸦retinal ganglion cell: RGC㸧ࢆྵࡴ⥙ ⭷ෆࡢ⚄⤒⣽⬊ࢿࢵࢺ࣮࣡ࢡࡼࡗ࡚ฎ⌮ࠊ⤫ྜࡉࢀࡿࠋࡲࡓࠊ⥙⭷ෆࡢ⚄⤒ᅇ㊰ࡣᬯ ᡤ࡛ࡢどぬሗࡢఏ㐩ࢆᢸ࠺ᮞయ⤒㊰ࠊ᫂ᡤ࡛ࡢどぬሗࡢఏ㐩ࢆᢸ࠺㗹య⤒ ㊰ࡀᏑᅾࡋࠊ୧⪅ࡣ┦ᙳ㡪ࢆཬࡰࡋྜࡗ࡚࠸ࡿࡇࡀ▱ࡽࢀࡿࠋࡇࡢࡼ࠺ࠊ✀ࠎࡢ ⚄⤒⣽⬊ࡀ」㞧࡞ࢿࢵࢺ࣮࣡ࢡࢆᙧᡂࡍࡿ⥙⭷ࡣࠊどぬሗࡢⓎ⏕ཬࡧఏ㐩ྍḞࡢ⤌ ⧊࡛࠶ࡿࠋ ୍᪉ࠊ⥙⭷ࡣ⸆࠶ࡿ࠸ࡣᏛ≀㉁ࡢᙳ㡪ࢆཷࡅࡸࡍ࠸⤌⧊࡛ࡶ࠶ࡿࠋࡑࡢ⌮⏤ࡋ࡚ ࡣࠊ⬦⤡⭷⾑ὶ㔞ࡀᴟࡵ࡚ከ࠸ࡇࠊ⬦⤡⭷ẟ⣽⾑⟶ࡀ᭷❆ᆺ࡛࠶ࡿࡇࠊ⬦⤡⭷ཬࡧ⥙ ⭷Ⰽ⣲ୖ⓶࣓ࣛࢽࣥࡀᏑᅾࡍࡿࡇࠊど⣽⬊࡛ࡢ࣑ࢺࢥࣥࢻࣜࡢ㓟ⓗ௦ㅰάᛶࡀ㧗 ࠸ࡇࠊど⣽⬊እ⠇ࡢ௦ㅰᅇ㌿ࡀ㧗࠸ࡇࠊ⥙⭷ෆከᵝ࡞⚄⤒ఏ㐩ཬࡧ⚄⤒ㄪ⠇≀㉁ࡀ Ꮡᅾࡍࡿࡇ➼ࠊᩘከࡃࡀᣲࡆࡽࢀࡿ(14)ࠋࡇࡢ⥙⭷ࡢ⸆ࡼࡿ㞀ᐖࠊࡘࡲࡾ⸆ㄏⓎᛶ ⥙⭷㞀ᐖࡣどぬࡢ␗ᖖࡘ࡞ࡀࡿࠋどぬࡣ♫ⓗ⏕άࢆႠࡴୖ࡛ᴟࡵ࡚㔜せ࡞ᶵ⬟࡛࠶ࡾࠊ ୍⯡ⓗࣄࢺࡣሗࡢ⣙ 80%ࢆどぬࡽᚓ࡚࠸ࡿゝࢃࢀࡿࠋࡘࡲࡾࠊ⸆ㄏⓎᛶ⥙⭷㞀2
Requirements for Registration of Pharmaceuticals for Human Use: ICH㸧ࡢྜពᇶ࡙ࡁసᡂࡉࢀ ࡓ༢ᅇཬࡧᢞẘᛶヨ㦂࢞ࢻࣛࣥ㸦S4㸧ࡣ་⸆ရࡢ〇㐀㈍ᢎㄆ⏦ㄳ㝿ࡋ࡚ ᐇࡍࡁ୍⯡ẘᛶヨ㦂࡛ࡢホ౯㡯┠ཬࡧࡑࡢ᪉ἲࡀᐃࡵࡽࢀ࡚࠾ࡾࠊ⥙⭷ࡢホ౯ࡶྠ࢞ ࢻࣛࣥ‽ࡌ࡚ᐇࡉࢀࡿࠋྠ࢞ࢻ࡚ࣛࣥホ౯ࡀ⩏ົࡅࡽࢀ࡚࠾ࡾ⥙⭷㞀ᐖ 㛵㐃ࡢ࠶ࡿホ౯㡯┠ࡋ࡚ࡣࠊ୍⯡≧ែࡢほᐹࠊ║⛉Ꮫⓗ᳨ᰝࠊཬࡧ⌮⤌⧊Ꮫⓗ᳨ᰝ ࡀᣲࡆࡽࢀࡿࠋ⥙⭷ࡢホ౯ࡣᙧែᏛⓗホ౯ἲᶵ⬟ⓗホ౯ἲู࡛ࡁࡿࠋ㠀⮫ᗋᏳᛶ ヨ㦂࠾ࡅࡿ⥙⭷ࡢᙧែᏛⓗホ౯ࡣ⸆ࡢᢞ㛤ጞ๓ཬࡧᢞᮇ㛫୰ࡢ᳨║㙾ࢆ⏝࠸ࡓ║ ᗏ᳨ᰝཬࡧゎ๗ᚋࡢ⌮⤌⧊Ꮫⓗ᳨ᰝࡀヱᙜࡋࠊ୧᳨ᰝࡽ⸆ࡼࡿ⥙⭷ࡢᙧែኚ 㛵ࡍࡿከࡃࡢሗࢆᚓࡿࡇࡀྍ⬟࡛࠶ࡿࠋ୍᪉ࠊᶵ⬟ⓗホ౯ࡋ࡚ࡣࠊ୍⯡≧ែほᐹ ࡼࡗ࡚どぬࡢ␗ᖖ㛵㐃ࡋࡓ⾜ືࡀほᐹࡉࢀࡿྍ⬟ᛶࡀ࠶ࡿࠋࡋࡋࠊ୍⯡ⓗື≀࡛ࡣ ⾜ືほᐹࡽ㍍ᗘࡢどぬ␗ᖖࢆ᳨ฟࡍࡿࡇࡣᴟࡵ࡚ᅔ㞴࡛࠶ࡿࠋࡉࡽࠊ⾜ືほᐹࡣ⥙ ⭷ࡽ⬻どぬ㔝⮳ࡿࡲ࡛ࡢどぬఏᑟ㊰ࡢ⥲యⓗホ౯࡛࠶ࡿࡓࡵࠊ⥙⭷ᶵ⬟ࢆ┤᥋ホ౯ ࡋ࡚࠸ࡿࡣゝ࠼࡞࠸ࠋࡑࡢࡓࡵࠊྠ࢞ࢻ࡛ࣛࣥࡶࠊᚲせᛂࡌ࡚どぬ㛵ࡍࡿᶵ⬟ ᳨ᰝࢆู㏵ᐇࡍࡿࡇࡀ᥎ዡࡉࢀ࡚࠸ࡿࠋᐇ㝿ࠊ㢮ఝ⸆ࡢሗࠊ⸆⌮స⏝ࠊᏛⓗᵓ 㐀➼ࡽ⥙⭷ᶵ⬟㞀ᐖࡀᠱᛕࡉࢀࡿሙྜࠊ⥙⭷ᶵ⬟ࡢ㟁Ẽ⏕⌮Ꮫⓗ࡞ホ౯᪉ἲ࡛࠶ࡿ⥙⭷ 㟁ᅗ㸦electroretinogram: ERG㸧ࡀࡋࡤࡋࡤ⏝࠸ࡽࢀࡿ(45)ࠋ ERG ࡣග่⃭ࡼࡗ࡚ㄏⓎࡉࢀࡿ⥙⭷ࡢ㟁ኚࢆグ㘓ࡋࡓࡶࡢ࡛࠶ࡿࠋග่⃭ࡼ ࡾ║⌫Ⓨ⏕ࡍࡿ⣽⬊እ㟁ὶࡢᴫᛕᅗࢆFig. 3 ♧ࡍࠋ୍⯡ⓗ࡞ ERG ࡣࠊྠᅗ࡚ B ࡛⾲ ࡋࡓᅇ㊰ࠊࡍ࡞ࢃࡕ◪Ꮚయཬࡧゅ⭷ࢆ⤒࡚⥙⭷ࢆ㞳ࢀࡓᚋ⬦⤡⭷ཬࡧ⥙⭷Ⰽ⣲ୖ⓶ࢆ⤒ ࡚⥙⭷ᡠࡿᅇ㊰ἢࡗ࡚グ㘓ࡉࢀࡿࠋERG ࡢἼᙧࡣ㡰ᛂ᮲௳ཬࡧග่⃭ࡢᙉᗘ➼ࡢグ㘓 ᮲௳ࡼࡗ࡚ࡁࡃኚࡍࡿࡀࠊ⥙⭷ᶵ⬟ࡢ␗ᖖࡢ᭷↓ࢆㄪࡿ┠ⓗ࡛ࡣࠊᬯᡤ୍࡚ᐃ ௨ୖࡢ㛫㡰ᛂࡋࡓ≧ែ࡛࠶ࡿᬯ㡰ᛂୗ࡚୰➼ᗘࡢᙉᗘࡢ㛝ග่⃭ࡼࡗ࡚ㄏⓎࡉࢀࡿ ᛂ⟅ࡀグ㘓ࡉࢀࡿࡇࡀ୍⯡ⓗ࡛࠶ࡿࠋFig. 4 ྠᛂ⟅ࡢᆺࢆ♧ࡍࠋྠᛂ⟅࡛ࡣග่⃭ ࡢ┤ᚋ㝜ᛶἼࡀࠊ⥆࠸࡚㝧ᛶἼࡀほᐹࡉࢀࠊࡑࢀࡒࢀa Ἴཬࡧ b Ἴࡤࢀࡿࠋ୍⯡ⓗࠊ ERG ࡢ a Ἴࡣ⥙⭷ࡢ୍ḟࢽ࣮࡛ࣗࣟࣥ࠶ࡿど⣽⬊ࠊb Ἴࡣḟࢽ࣮࡛ࣗࣟࣥ࠶ࡿᴟ⣽⬊
ࠊࡑࢀࡒࢀࡋࡓ㉳※ࢆ᭷ࡍࡿ⪃࠼ࡽࢀ࡚࠸ࡿࠋ ࣄࢺࡢ⮫ᗋ࡛ࡣࠊERG ࡀ⥙⭷ᶵ⬟ࢆぬⓗホ౯ࡍࡿࡇࡢ࡛ࡁࡿ㔜せ࡞᳨ᰝ⨨ ࡅࡽࢀ࡚࠸ࡿࠋ୰㛫㏱ගయΰ⃮࠶ࡿ࠸ࡣᑠඣࡢ⥙⭷ᶵ⬟ุᐃࠊ⥙⭷⾑⟶␗ᖖཬࡧ⥙⭷ ⾑ࡢホ౯ࠊཎᅉ᫂ࡢどᶵ⬟㞀ᐖࡢデ᩿➼ࠊከࡃࡢሙ㠃࡛ERG ࡀά⏝ࡉࢀ࡚࠸ࡿࠋࡇࢀ ᑐࡋ࡚ື≀ᐇ㦂ࡢ㡿ᇦ࡛ࡣࠊERG ࡣ⮬ぬⓗ࡞᳨ᰝࡀᅔ㞴࡞ື≀ࣔࢹ࡛ࣝࡢどぬᶵ⬟ࡢホ౯ ⏝࠸ࡽࢀ࡚࠸ࡿࠋERG ࡀどぬࡢホ౯⏝࠸ࡽࢀࡿື≀✀ࡣከᒱࢃࡓࡾࠊ࣓ࢲ࢝ࠊ ࣐࢘ࢫࠊࣛࢵࢺࠊࣁ࣒ࢫࢱ࣮ࠊ࢘ࢧࢠࠊࢿࢥࠊࢾࠊࣈࢱࠊཬࡧࢧࣝ࡞ᐇᵝࠎ࡛࠶ࡿ(6, 53)ࠋࡲࡓࠊERG ࡢἼᙧࡣ✀ᕪࡀ࠶ࡾࠊࡇࢀࡣ⥙⭷ࡢゎ๗Ꮫⓗཬࡧ⏕⌮Ꮫⓗᕪ␗ࢆᫎࡋ ࡓࡶࡢ⪃࠼ࡽࢀࡿࠋ࠼ࡤࠊど⣽⬊༨ࡵࡿ㗹యࡢྜࡀࣄࢺẚ㍑ࡋ࡚ᑠࡉ࠸ࡆࡗṑ 㢮࡛ࡣࠊ㗹య⤒㊰ࡢᶵ⬟ࢆᫎࡍࡿ ERG ࡛࠶ࡿ㗹య⣔ᛂ⟅㸦ᚋ㏙㸧 a Ἴࡀࢇㄆࡵ ࡽࢀ࡞࠸ࠋࡉࡽࠊERG ࡢἼᙧᡂศࡢ㉳※ࡶ✀ᕪࡀ▱ࡽࢀࡿ(37)ࠋࡑࡢࡓࡵࠊື≀ᐇ㦂 ࡚ERG ࢆグ㘓ࡍࡿ㝿ࡣࠊᐇ㦂┠ⓗྜ⮴ࡋࡓື≀✀ࢆ㑅ᢥࡍࡿࡇࡀ㔜せ࡛࠶ࡿࠋ 㠀⮫ᗋᏳᛶヨ㦂࡚⥙⭷㞀ᐖࢆホ౯ࡍࡿୖ࡛ࡣࠊ௨ୗࡢ 3 ࡘࡢⅬࡼࡾ࢝ࢽࢡࢨ ࣝࡀ᭱ࡶ㐺ษ࡞ື≀✀ࡔ⪃࠼ࡽࢀࡿࠋ➨୍ࡢⅬࡋ࡚ࠊࢧࣝࡢ║ࡣゎ๗Ꮫⓗࣄࢺ ᴟࡵ࡚㢮ఝࡋ࡚࠸ࡿࡇࡀᣲࡆࡽࢀࡿࠋ࠼ࡤࠊࢧࣝࡢ⥙⭷࡛ࡣࣄࢺࡢࡼ࠺㗹యࡢᐦ㞟 ࡋࡓ㯤ᩬࡀⓎ㐩ࡋ࡚࠾ࡾࠊど⣽⬊ࡢᮞయ㗹యࡢẚ⋡ཬࡧศᕸࡀࢧࣝࣄࢺ࡛㏆࠸ࡇࡀ ▱ࡽࢀ࡚࠸ࡿ(10, 75)ࠋࡇࡢ㧗࠸㢮ఝᛶࡣࠊヨ㦂⤖ᯝࢆࣄࢺእᤄࡍࡿ㝿᭷࡛࠶ࡿࡇࢆ ពࡍࡿࠋ࠼ࡤࠊ⥙⭷ẘᛶࡘ࠸࡚᭱ࡶᗈ⠊ᅖ◊✲ࡉࢀࡓ⸆ࡢ୍ࡘࢡࣟࣟ࢟ࣥࡀ
࠶ࡿ㸦ࢡࣟࣟ࢟ࣥ⥙⭷㸧ࠋࢡࣟࣟ࢟ࣥ⥙⭷ᝈ⪅ࡢ║ᗏീ࡛ࡣbull’s eye maculopathy
ࡤࢀࡿ≉ᚩⓗ࡞ኚࡀ㯤ᩬ㒊ㄆࡵࡽࢀࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ(67)ࠋࡋࡋࠊ㠀⮫ᗋᏳᛶ
ヨ㦂⏝࠸ࡽࢀࡿື≀✀ࡢ࠺ࡕ㯤ᩬࢆᣢࡓ࡞࠸࣐࢘ࢫࠊࣛࢵࢺ࠶ࡿ࠸ࡣࢾ࠾࠸࡚ྠ ኚࢆ⌧ࡍࡿࡇࡣྍ⬟࡛࠶ࡿࠋ➨ࡢⅬࡣࠊ࢝ࢽࢡࢨࣝࢆྵࡴ࣐࢝ࢡᒓ࡛ࡣࠊࡑ
4 ࡀ㠀⮫ᗋᏳᛶヨ㦂୍⯡ⓗ⏝࠸ࡽࢀࡿື≀✀ࡢ୍ࡘ࡛࠶ࡿࡇࡀᣲࡆࡽࢀࡿࠋIn vivo ホ౯⣔ࡋ࡚ࠊヨ㦂⏝࠸ࡿື≀ࡀᏳᐃⓗ౪⤥ࡉࢀ࡚࠸ࡿࡇࡣ㔜せ࡛࠶ࡿࠋࡉࡽࠊ ᢞẘᛶヨ㦂➼ホ౯⣔ࢆ⤌ࡳ㎸ࡴࡇࡀྍ⬟࡞ࡿࡓࡵࠊྠ୍ヨ㦂ෆ࡛ᚓࡽࢀࡿ᭚ 㟢ሗࡸࡢ᳨ᰝ⤖ᯝྜࢃࡏ࡚ࣄࢺ࡛ࡢ⥙⭷ᶵ⬟㞀ᐖࡢࣜࢫࢡࢆࡼࡾ㐺ษホ౯ࡍࡿࡇ ࡘ࡞ࡀࡿ⪃࠼ࡽࢀࡿࠋ୍᪉ࠊࡆࡗṑ㢮ཬࡧࢾẚࡿࣁࣥࢻࣜࣥࢢ༴㝤ࡀక ࠺ࡇࡀࢧࣝ㢮ࢆᐇ㦂⏝࠸ࡿࡇࡢḞⅬࡋ࡚ᣲࡆࡽࢀࡿࡀࠊࡇࡢ༴㝤ࡣヨ㦂ᚑ⪅ࡢ 㐺ษ࡞ᩍ⫱ࡼࡗ࡚ᖜ㍍ῶࡍࡿࡇࡀྍ⬟࡛࠶ࡿࠋࡇࢀࡽࡢࡇࡽࠊ࢝ࢽࢡࢨࣝ ࡛ࡢ☜❧ࡉࢀࡓERG ࡢグ㘓ཬࡧゎᯒἲࡣࠊ㠀⮫ᗋᏳᛶヨ㦂࡚⥙⭷ᶵ⬟㞀ᐖࢆ᳨ฟཬࡧ ࣔࢽࢱࣜࣥࢢࡋࠊࡉࡽ⥙⭷୰࡛ࡢ⸆ࡢస⏝㒊ࢆヲ⣽ㄪࡿᡭἲࡋ࡚㠀ᖖ᭷⏝ ࡛࠶ࡿࡇࡀᮇᚅࡉࢀࡿࠋ ㏆ᖺࠊ㠀⮫ᗋᏳᛶヨ㦂࠾ࡅࡿࢧࣝࡢERG ࡀὀ┠ࡉࢀࠊࡑࡢ᪉ἲࡢᶆ‽ࡶヨࡳࡽࢀ ጞࡵࡓࠋ(2)ࡋࡋࠊ⸆ㄏⓎᛶ⥙⭷ᶵ⬟㞀ᐖࡢホ౯⣔ࡋ࡚ࡢ࢝ࢽࢡࢨ࡛ࣝࡢ ERG グ㘓 ཬࡧゎᯒἲ㝈ᐃࡍࡿࠊࡇࢀࡲ࡛ࡢࡇࢁሗࡣᴟࡵ࡚ᑡ࡞࠸ࠋ࠼ࡤࠊࣄࢺ⮫ᗋ࡛ࡣ ⥙⭷ෆࡢᮞయ⤒㊰㗹య⤒㊰⏤᮶ࡍࡿERG ࢆศ㞳ࡋ࡚グ㘓ࡍࡿᶆ‽ࡉࢀࡓᡭἲ(44)ࡀ☜ ❧ࡉࢀ࡚࠸ࡿࡀࠊ⸆➼ࢆᢞࡋࡓ࢝ࢽࢡࢨ࡛ࣝᮞయ⤒㊰㗹య⤒㊰ࢆ⪃៖ࡋࡓERG ࢆ グ㘓ࡋࡓሗ࿌ࡣ㠀ᖖᑡ࡞࠸ࠋ(19, 29)ࡲࡓࠊ࢝ࢽࢡࢨ࡛ࣝࡢERG ࢆ⏝࠸࡚⥙⭷ෆࡢ≉ᐃࡢ ⚄⤒⣽⬊ཬࡰࡍ⸆ࡢᙳ㡪ࢆㄪࡓሗ࿌ࡣࠊⴭ⪅ࡢ▱ࡿ㝈ࡾ࡞࠸ࠋࡇࢀࡽࡢࡇࡽࠊ 㠀⮫ᗋᏳᛶศ㔝࠾࠸࡚ࠊ࢝ࢽࢡࢨ࡛ࣝࡢERG グ㘓ࡀ⥙⭷㞀ᐖࡢࣜࢫࢡホ౯༑ศ ά⏝ࡉࢀ࡚࠸࡞࠸ࡢࡀ⌧≧࡛࠶ࡿࠋࡑࡇ࡛ᮏ◊✲࡛ࡣࠊ࢝ࢽࢡࢨࣝ࠾ࡅࡿ⸆ㄏⓎᛶ ⥙⭷ᶵ⬟㞀ᐖࡢయ⣔ⓗ࡞ホ౯ἲࡢᵓ⠏ࢆ┠ⓗࡋ࡚ࠊࣄࢺ࠶ࡿ࠸ࡣ࣐࢝ࢡᒓ࡛ࡢ᪤ሗࢆཧ ⪃ࠊ⥙⭷ࡢྛᵓᡂᡂศ↔Ⅼࢆᙜ࡚ࡓྛ✀ERG ࡢグ㘓ཬࡧゎᯒ᪉ἲࢆ᳨ウࡋࡓࠋ➨୍❶ ࡛ࡣࠊ⥙⭷ෆࡢ⤒㊰ࡋ࡚2 ࡘูࡉࢀࡿᮞయ⤒㊰ཬࡧ㗹య⤒㊰ࡢᶵ⬟ホ౯╔┠ࡋࡓࠋ ➨㹼ᅄ❶࡛ࡣࠊࡑࢀࡒࢀ⥙⭷ෆࡢ୍ࠊࠊཬࡧ୕ḟࢽ࣮࡛ࣗࣟࣥ࠶ࡿど⣽⬊ࠊᴟ⣽⬊ࠊ ཬࡧRGC ࡢᶵ⬟ホ౯╔┠ࡋࡓࠋ
6
FIGURE 2. (A) A photomicrograph of a vertical section through the central retina in a
cynomolgus monkey and (B) diagram of the corresponding nerve circuit of the retina. ILM, inner limiting membrane; NFL, nerve fiber layer; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; OLM, outer limiting membrane; RPE, retinal pigment epithelium.
FIGURE 3. A schematic representation of the extracellular currents that are formed following
light stimulation. The pathway “A” represents local currents within the retina, while pathway “B” shows the currents leaving the retina through the vitreous and the cornea and returning to the retina through the choroid and the pigment epithelium. ERG recording is done along the pathway B.
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FIGURE 4. A typical waveform of the ERG response elicited by the white light flash of moderate
intensity in a dark-adapted cynomolgus monkey. Arrowhead indicates onset of the light flash. Two components that are most often measured are the a- and b-waves. The a-wave is the first large negative component, followed by the b-wave which is corneal positive and usually larger in amplitude. Two principal measures of the ERG waveform are taken: 1) The amplitude from the baseline to the negative trough of the a-wave, and the amplitude of the b-wave measured from the trough of the a-wave to the following peak of the b-wave; and 2) the time from flash onset to the trough of the a-wave and the time from flash onset to the peak of the b-wave. It is well known that the waveform of ERGs recorded in cynomolgus monkeys is quite similar to that recorded in humans.
➨
1 ❶ Standard full-field ERG㸸ᮞయ⤒㊰ཬࡧ㗹య⤒㊰ࡢศ㞳ホ౯
1. ᗎㄽ ⥙⭷ෆࡢගཷᐜయ࡛࠶ࡿど⣽⬊ࡣࠊග㔞ࡀᑡ࡞࠸≧ἣ࡛ࡢどぬࠊࡍ࡞ࢃࡕᬯᡤどࢆᢸ ࠺ᮞయࠊග㔞ࡀศ࠶ࡿ≧ἣ࡛ࡢどぬࠊࡍ࡞ࢃࡕ᫂ᡤどࢆᢸ࠺㗹యࡢࡘࡢࢱࣉࡀ Ꮡᅾࡍࡿࠋࡲࡓࠊ⥙⭷ෆ࡛ᮞయ⏤᮶ࡍࡿどぬሗࡢఏᑟ㊰ࡣᮞయ⤒㊰ࠊ㗹య⏤᮶ࡍࡿ どぬሗࡢఏ㐩㊰ࡣ㗹య⤒㊰ࡑࢀࡒࢀࡤࢀࡿࠋERG ࡛ࡣࠊගᑐࡍࡿឤᗘ࠶ࡿ࠸ࡣ 㛫ศゎ⬟➼ࠊᮞయ㗹య࠾ࡅࡿ⏕⌮Ꮫⓗ≉ᛶࡢᕪ␗ࢆ⏝ࡋࠊᮞయ⤒㊰ཬࡧ㗹య⤒㊰ࡢ ᛂࢆࡑࢀࡒࢀᮞయ⣔ᛂ⟅ཬࡧ㗹య⣔ᛂ⟅ࡋ࡚ศ㞳ࡋ࡚グ㘓ࡍࡿࡇࡀྍ⬟࡛࠶ࡿࠋࡇ ࢀࡼࡗ࡚ࠊ⥙⭷ෆ࡛ᶵ⬟␗ᖖࡀᏑᅾࡍࡿ㒊ࢆࡼࡾṇ☜ᢕᥱࡍࡿࡇࡀ࡛ࡁࡿࠋ ࣄࢺ⮫ᗋ࡛ࡣࠊERG ࡣ⥙⭷ᶵ⬟ࢆぬⓗホ౯ࡍࡿࡇࡢ࡛ࡁࡿ㔜せ࡞᳨ᰝ⨨ࡅ ࡽࢀ࡚࠸ࡿࠋ㗹యࢪࢫࢺࣟࣇ࣮ࡸඛኳᛶኪ┣➼ࡢ⥙⭷ᝈࡣࠊࡑࡢデ᩿ୖグࡢᮞయ⤒ ㊰㗹య⤒㊰ࡢᛂࢆࡑࢀࡒࢀศ㞳ࡋ࡚グ㘓ࡋࡓ ERG ࡀḞࡃࡇࡀ࡛ࡁ࡞࠸ࠋࡑࡢ㠃ࠊ ࣄࢺ⮫ᗋ࡛ࡣྛタࡀ⊂⮬タᐃࡋࡓ᮲௳ࡼࡗ࡚ERG ࡀグ㘓ࡉࢀ࡚ࡁࡓ⤒⦋ࡀ࠶ࡾࠊࡇ ࡢࡓࡵ㛗ࡽࡃタ㛫ࡢERG ࡢẚ㍑ࡀᅔ㞴࡛࠶ࡗࡓࠋࡇࡢၥ㢟ࢆゎỴࡍࡿࡓࡵࠊ1989 ᖺᅜ㝿⮫ᗋどぬ㟁Ẽ⏕⌮Ꮫ㸦International Society for Clinical Electrophysiology of Vision: ISCEV㸧 ࡀࠊࣄ ࢺ⮫ᗋ࡚ᮞయ⤒ ㊰㗹య⤒㊰ࡢ ᛂࢆศ㞳ࡋ࡚グ㘓 ࡍࡿᶆ‽ ࣉࣟࢺ ࢥ࣮ࣝ
㸦ISCEV ࣉࣟࢺࢥ࣮ࣝ㸧ࢆᥦࡋࡓ(40)ࠋ⌧ᅾࠊISCEV ࣉࣟࢺࢥ࣮ࣝࡣࣄࢺ⮫ᗋ࡚ ERG ࢆ
グ㘓ࡍࡿ㝿ࡢᶆ‽ⓗ࡞᪉ἲࡋ࡚ᐃ╔ࡋ࡚࠾ࡾࠊྠࣉࣟࢺࢥ࣮ࣝ‽ᣐࡋ࡚グ㘓ࡉࢀࡓ ERG ࡣ standard full-field ERGs ࡤࢀࡿ(41-44)ࠋ
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㟁Ẽ⏕⌮Ꮫⓗ᳨ᰝẚ㍑ࡍࡿ㠀⮫ᗋᏳᛶศ㔝࡛ERG ࢆ⇍▱ࡋࡓ◊✲⪅ࡀᑡ࡞࠸ࡇ
ࡽࠊࡇࢀࡲ࡛ᚓࡽࢀ࡚࠸ࡿሗࡀᑡ࡞ࡃࠊstandard full-field ERG ࡢ᭷⏝ᛶࡀ༑ศ⌮ゎࡉࢀ ࡚࠸࡞࠸ࡇࡀᣲࡆࡽࢀࡿࠋࡋࡋ࡞ࡀࡽࠊ⥙⭷ᑐࡋ࡚ఱࡽࡢᶵ⬟ⓗᙳ㡪ࢆཬࡰࡍࡇ ࡀ᫂ࡽ࡞ྜ≀ࢆ་⸆ရࡋ࡚㛤Ⓨࡍࡿሙྜࠊ㠀⮫ᗋᏳᛶヨ㦂࡚standard full-field ERGs ࢆグ㘓ࡍࡿࡇࡼࡾ᭷⏝࡞ሗࡀᚓࡽࢀࡿ⪃࠼ࡽࢀࡿࠋࡍ࡞ࢃࡕࠊᚓࡽࢀࡓ ERG ࡢኚࡽࣄࢺ࡛ࡢどぬ㞀ᐖࡢ㉁ࢆண ࡋࠊࡉࡽ⮫ᗋヨ㦂ྠ୍ࡢ࣐࣮࣮࡚࢝㠀⮫ᗋ Ᏻᛶヨ㦂࡛Ⓨ⌧ࡍࡿᶵ⬟㞀ᐖࢆࣔࢽࢱࣜࣥࢢࡍࡿࡇࡀྍ⬟࡞ࡿ⪃࠼ࡽࢀࡿࠋ ୍᪉ࠊࢺࣜࢰ࣮ࣝ⣔ᢠ┿⳦࡛࠶ࡿ࣎ࣜࢥࢼࢰ࣮ࣝࡣࠊࣄࢺᑐࡋ࡚ᢞᚋ⩈᫂ࠊ 㟝どࠊⰍぬࡢኚ➼ࡢどぬ㞀ᐖࢆ୍㐣ᛶㄏⓎࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ(11, 12, 18, 55, 66)ࠋ࣎ ࣜࢥࢼࢰ࣮ࣝࡢ⮫ᗋヨ㦂࡛ࡣࠊᖖே⤒ཱྀᢞࡋࡓ㝿ࡢ⥙⭷ᶵ⬟ࡢኚࡀ standard full-field ERGs ࡚ホ౯ࡉࢀࡓ(12)ࠋࡇࡢヨ㦂࡛ࡣࠊᮞయ⤒㊰㗹య⤒㊰ࡢ୧⤒㊰ࡢᶵ⬟ࢆ ᫎࡍࡿΰྜᛂ⟅࡛᭷ព࡞ᖜపୗࡀ᳨ฟࡉࢀࡓ୍᪉࡛ࠊ㗹య⤒㊰ࡢᶵ⬟ࢆᫎࡍࡿ㗹య⣔ ᛂ⟅ࡣ㢧ⴭ࡞ኚࡣほᐹࡉࢀ࡞ࡗࡓࠋࡇࡢ⤖ᯝࡽࠊࣄࢺ࠾ࡅࡿ࣎ࣜࢥࢼࢰ࣮ࣝ ࡼࡿ⥙⭷ᶵ⬟ࡢᙳ㡪ࡀᮞయ⤒㊰࡛㢧ⴭ࡛࠶ࡗࡓࡇࡀ♧၀ࡉࢀࡿࠋࡲࡓࠊࣄࢺ⮫ᗋ࡚ ࠺ࡗ⾑ᛶᚰࡢ⒪⸆ࡋ࡚⏝ࡉࢀ࠸ࡿࢪࢦ࢟ࢩࣥࡣࠊ୰ẘᝈ⪅どຊపୗࠊ㟝どࠊ 㯤ど➼ࡢどぬ␗ᖖࢆྍ㏫ⓗㄏⓎࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ(8, 39, 49, 54, 74)ࠋሗ࿌(49, 74)ࡼࡿ ࠊ୰ẘࡢ ERG ࡢ␗ᖖࡋ࡚ࠊ㗹య⣔ᛂ⟅ࡢྍ㏫ⓗ࡞ῶᙅཬࡧ㐜ᘏࡀㄆࡵࡽࢀ࡚࠾ࡾࠊ 㗹య⤒㊰ࡢᶵ⬟㞀ᐖࡀᙉࡃ♧၀ࡉࢀࡿࠋ୍᪉ࠊྠሗ࿌࠾ࡅࡿᮞయ⣔ᛂ⟅ࡣࠊ୰ẘࡣ ṇᖖ⠊ᅖෆ࡛࠶ࡗࡓࡶࡢࡢఇ⸆ࡼࡾ₯▷⦰࠶ࡿ࠸ࡣῶᙅࡀㄆࡵࡽࢀࡓࡇࡽࠊ୰ẘ ࡢᮞయ⤒㊰ࡢᛂࡣࡴࡋࢁቑഴྥࢆ♧ࡋ࡚࠸ࡓ᥎ᐹࡉࢀࡿࠋ ᮏ◊✲࡛ࡣࠊ㠀⮫ᗋᏳᛶヨ㦂୍⯡ⓗ⏝࠸ࡽࢀࡿື≀✀࡛࠶ࡿ࢝ࢽࢡࢨࣝࢆᑐ㇟ ࡋࠊISCEV ࣉࣟࢺࢥ࣮ࣝࢆཧ⪃ standard full-field ERGsࠊࡍ࡞ࢃࡕࠊᮞయ⣔ᛂ⟅㸦rod response㸧ࠊΰྜᛂ⟅㸦combined rod-cone response㸧ཬࡧ㗹య⣔ᛂ⟅㸦single-flash cone response
ཬࡧ30 Hz flicker㸧ࡢグ㘓᮲௳ࢆタᐃࡋࡓࠋࡲࡓࠊ๓㏙ࡢࡼ࠺ࣄࢺ࡛ࡑࢀࡒࢀᮞయ⤒㊰ཬ
ࢪࢦ࢟ࢩࣥࡢస⏝ࡀࠊ࢝ࢽࢡࢨ࡛ࣝࡢstandard full-field ERGs ࡼࡗ᳨࡚ฟྍ⬟࡛࠶ࡿ ࢆ᳨ウࡋࡓࠋ 2. ᐇ㦂ᮦᩱཬࡧᐇ㦂᪉ἲ ᮏᐇ㦂ࡣࠊ➨୍୕ඹᰴᘧ♫ࡢື≀ᐇ㦂ጤဨࡢᢎㄆ㸦ື≀ᐇ㦂ᢎㄆ␒ྕ㸸C800264 ཬࡧ C800044㸧ࢆᚓ࡚ᐇࡋࡓࠋ 2. ື≀ 2.1. ື≀ ୍⯡≧ែཬࡧ║ᗏ␗ᖖࡢ࡞࠸㞤㞝ྛ 6 ࠊィ 12 ࡢ୰ᅜ⏘࢝ࢽࢡࢨࣝ㸦3㸫5 ṓ㱋ࠊ య㔜2.8㸫3.8 kg㸧ࢆ⏝࠸ࡓࠋ㞤㞝ྛ 3 ࠊィ 6 ࡣ࣎ࣜࢥࢼࢰ࣮ࣝᢞ⩌㸦3㸫4 ṓ㱋ࠊయ 㔜2.8㸫3.5 kg㸧ࠊṧࡾࡢ 6 ࡣࢪࢦ࢟ࢩࣥᢞ⩌㸦3㸫4 ṓ㱋ࠊయ㔜 2.8㸫3.8 kg㸧ࡾ ศࡅࡓࠋື≀ࡣERG ᐇ㦂୰ࢆ㝖ࡁࠊ ᗘ 24Υࠊ‵ᗘ 60%ࠊ↷᫂ 12 㛫㸭᪥㸦7:00㸫19:00㸧ࠊ ↷ᗘ 150㸫300 ࣝࢡࢫࠊẼᅇᩘ 10㸫15 ᅇ㸭㛫タᐃࡋࡓヨ㦂ᐊ࡚ࠊࢫࢸࣥࣞࢫ〇ࢣ ࣮ࢪෆಶู㣫⫱ࡋࡓࠋ⤥䭱ࡣࠊࢧࣝ⏝ᅛᙧ㣫ᩱ㸦PS㸹࢚࢜ࣜࣥࢱࣝ㓝ẕᕤᴗᰴᘧ♫ࠊ ᮾி㸧ࢆ1 ᪥ 1 ᅇ 100 g ࠼ࡓࠋ⤥ỈࡣࠊỈ㐨Ỉࢆ⤥Ỉࣀࢬࣝࡽ⮬⏤ᦤྲྀࡉࡏࡓࠋ 2.2. ᢞ ࣄࢺ࡛ᮞయ⤒㊰ᙉ࠸ᙳ㡪ࢆཬࡰࡍࡇࡀ▱ࡽࢀ࡚࠸ࡿ⸆ࡋ࡚ࠊ࣎ࣜࢥࢼࢰ࣮ࣝࢆ
㑅ᢥࡋࡓࠋ࣎ࣜࢥࢼࢰ࣮ࣝ㸦ࣈࣇ࢙ࣥࢻ㟼ὀ⏝200 mg; Pfizer Inc., New York, NY, USA㸧ࡣ
⏕⌮㣗ሷᾮ࡚ᕼ㔘ࡋ࡚ᢞᾮࢆㄪ〇ࡋࡓࠋᢞᾮࡣࠊ6 ࡢື≀ 0㸦⏕⌮㣗ሷᾮ㸧ࠊ3ࠊ 6ࠊཬࡧ 12 mg/kg ࡢ⏝㔞ࢆྛ⏝㔞ࡀ༢ᅇᢞྠᵝࡳ࡞ࡏࡿ⪃࠼ࡽࢀࡿ 1 㐌㛫௨ୖࡢ㛫
12
ࣥࢪ࣏ࣥࣉ㸦22; Harvard Apparatus, Inc., Holliston, MA, USA㸧ࢆ⏝࠸࡚అᅾ㟼⬦ࡽ⾜ࡗࡓࠋ ࣄࢺ࡛㗹య⤒㊰ᙉ࠸ᙳ㡪ࢆཬࡰࡍࡇࡀ▱ࡽࢀ࡚࠸ࡿ⸆ࡋ࡚ࠊࢪࢦ࢟ࢩࣥࢆ㑅ᢥ ࡋࡓࠋࢪࢦ࢟ࢩࣥ㸦ࢪࢦࢩࣥὀ0.25 mg㸹୰እ〇⸆ᰴᘧ♫ࠊᮾி㸧ࡣ⏕⌮㣗ሷᾮ࡚ᕼ㔘 ࡋ࡚ᢞᾮࢆㄪ〇ࡋࡓࠋᢞᾮࡣࠊୖグࡣูࡢ6 0.1 mg/kg ࡢ⏝㔞࡚༢ᅇ㟼⬦ෆᢞ 㸦0.05 mL/kg/min40 min㸧ࡋࠊᢞ 28 ᪥ᚋࡲ࡛ほᐹࡋࡓࠋᢞ⏝㔞ࡣࠊࣄࢺ⮫ᗋ࡛ࡢ᥎ ዡ⏝㔞㸦0.125-0.25 mg/men/dayࠊ⤒ཱྀ㸧(65)ࣄࢺ࡛ࡢ⏕యෆ⏝⋡㸦66%㸧(60)ᇶ࡙ࡁࠊࣄ ࢺᝈ⪅ࢆୖᅇࡿ᭚㟢ࡀᚓࡽࢀࡿぢ㎸ࡲࢀࡓ⏝㔞ࡋ࡚タᐃࡋࡓࠋࢪࢦ࢟ࢩࣥࢆᢞࡍࡿ6 ᪥๓ࡣࠊ⏕⌮㣗ሷᾮࢆྠ 6 ྠᵝᢞࡋࡓࠋᢞࡣࠊࢩࣜࣥࢪ࣏ࣥࣉ㸦22㸹Harvard Apparatus, Inc.㸧ࢆ⏝࠸࡚అᅾ㟼⬦ࡽ⾜ࡗࡓࠋ 2.3. ERG 2.3.1. ๓ฎ⨨ ື≀ࡣࠊሷ㓟ࢣࢱ࣑ࣥ㸦ࢣࢱ࣮ࣛࣝ➽ὀ⏝500 mg㸹➨୍୕ඹᰴᘧ♫ࠊᮾி㸧㸦ᑟධ㸸10 mg/kgࠊ⥔ᣢ㸸5-10 mg/kg/h㸧ཬࡧ࢟ࢩࣛࢪࣥሷ㓟ሷ㸦ࢭࣛࢡࢱ࣮ࣝ㸹ࣂ࢚࣓ࣝࢹ࢝ࣝᰴ ᘧ♫ࠊ㜰㸧0.6 mg/kg ࡢ➽⫗ෆᢞࡼࡾ㯞㓉ࡋࡓࠋ▖Ꮝࡣࢺࣟࣆ࣑࢝ࢻཬࡧࣇ࢙ࢽࣞ ࣇࣜࣥሷ㓟ሷ㸦࣑ࢻࣜࣥ P Ⅼ║ᾮ㸹ཧኳ〇⸆ᰴᘧ♫ࠊ㜰㸧ࡢⅬ║ࡼࡾᩓ▖ࡉࡏࡓࠋ ゅ⭷ࡣሷ㓟࢜࢟ࢩࣈࣉࣟ࢝ࣥ㸦࣋ࣀ࢟ࢩ࣮ࣝⅬ║ᾮ0.4㸣㸹ཧኳ〇⸆ᰴᘧ♫㸧ࡢⅬ║ ࡼࡾ㯞㓉ࡋࡓᚋࠊࣄࢻࣟ࢟ࢩ࢚ࢳࣝࢭ࣮ࣝࣟࢫ㸦ࢫࢥࣆࢰࣝ║⛉⏝ᾮ㸹Ṋ⏣⸆ရᕤᴗᰴᘧ ♫ࠊ㜰㸧ࡢⅬ║ࡼࡾಖㆤࡋࡓࠋ 2.3.2. ග่⃭
ERG ࡢㄏⓎࡣ࢟ࢭࣀࣥࣛࣥࣉࢆග※ࡍࡿど㔝่⃭⨨㸦Ganzfeld Stimulator Model
2503S; LKC Technologies, Inc., Gaithersburg, MD, USA㸧ࢆ⏝ࡋࡓࠋど㔝่⃭⨨ࡢࢻ࣮࣒
⾲㠃࡚ᐇ ࡋࡓ่⃭᭱ᙉᗘࡣ6.3 cd∙s/m2࡛࠶ࡾࠊග่⃭ࡢᙉᗘࡣNeutral density㸦ND㸧
࠸࡚ῶᙅࡉࡏࡿࡇ࡛ㄪ⠇ࡋࡓࠋ
2.3.3. グ㘓ཬࡧゎᯒ
ISCEV ࡢ᥎ዡࡍࡿ᪉ἲ(44)ࢆཧ⪃ࠊⓑⰍࡢ่⃭ගཬࡧ⫼ᬒගࢆ⏝࠸࡚ standard full-field
ERGs ࢆグ㘓ࡋࡓࠋグ㘓㟁ᴟࡋ࡚ᴟᆺࢥࣥࢱࢡࢺࣞࣥࢬ㟁ᴟ㸦H6515NFC㸹᭷㝈♫࣓ ࣮ࣚࠊឡ▱㸧ࢆᕥ║ࠊ᥋ᆅ㟁ᴟࡋ࡚㔪㟁ᴟ㸦TN208-016㸹ᰴᘧ♫ࣘࢽ࣮ࢡ࣓ࢹ࢝
ࣝࠊᮾி㸧ࢆ㢌㡬㒊ࡢ⓶ୗࡑࢀࡒࢀタ⨨ࡋࡓࠋ40 ศ௨ୖࡢᬯ㡰ᛂᚋࠊ0.007 ཬࡧ 2.7 cd∙s/m2
ࡢᙉᗘࡢග่⃭࡚ࠊࡑࢀࡒࢀᮞయ⣔ᛂ⟅㸦rod response㸧ཬࡧΰྜᛂ⟅㸦combined rod-cone
response㸧ࢆㄏⓎࡋࡓࠋࡑࡢᚋࠊ40 cd/m2ࡢ㍤ᗘࡢ⫼ᬒග࡚10 ศ㛫ࡢ᫂㡰ᛂࢆ⾜࠸ࠊྠ⫼
ᬒගୗ࡚2.7 cd∙s/m2ࡢᙉᗘࡢග่⃭࡚㗹య⣔ᛂ⟅㸦single-flash cone response ཬࡧ 30 Hz
flicker㸧ࢆㄏⓎࡋࡓࠋୖグࡢ ERG ࡣࠊ࣎ࣜࢥࢼࢰ࣮ࣝࡢᢞ┤ᚋཬࡧᢞ 24 㛫ᚋࠊ࠶ࡿ ࠸ࡣࢪࢦ࢟ࢩࣥࡢᢞ๓㸦፹యࡢᢞ24 㛫ᚋ㸧ࠊᢞ┤ᚋࠊᢞ 24 㛫ࠊ7ࠊ14 ཬࡧ 28 ᪥ᚋࡑࢀࡒࢀグ㘓ࡋࡓࠋ ࡍ࡚ࡢᛂ⟅ࡣ 0.5㸫1000 Hz ࡢᖏᇦࣇࣝࢱ࣮ࢆ㏻ࡋ࡚ቑᖜࡋࡓᚋࠊㄏⓎ㟁᳨ᰝ ⨨㸦MEB-9104㸹᪥ᮏග㟁ᰴᘧ♫ࠊᮾி㸧グ㘓ࡋࡓࠋྠ୍᮲௳࡚グ㘓ࡉࢀࡓ 3 ࡽ 6 Ἴᙧࢆ⏝࠸࡚ຍ⟬ᖹᆒἼᙧࢆ⟬ฟࡋࡓࠋἼᙧᡂศࡢゎᯒࡋ࡚ࠊa Ἴࡢᖜ㸦ᇶ⥺ࡽ a Ἴ
ࢺࣛࣇࡲ࡛㸧ཬࡧ₯㸦ග่⃭ࡽ a Ἴࢺࣛࣇࡲ࡛㸧ࢆ combined rod-cone response ཬࡧ
single-flash cone response ࡢἼᙧࡽࠊb Ἴࡢᖜ㸦a Ἴࢺࣛࣇࡽ b Ἴ㡬Ⅼࡲ࡛㸧ཬࡧ₯
㸦ග่⃭ࡽb Ἴࣆ࣮ࢡࡲ࡛㸧ࢆ rod responseࠊcombined rod-cone responseࠊsingle-flash cone
responseࠊཬࡧ 30 Hz flicker ࡢἼᙧࡽࡑࢀࡒࢀィ ࡋࡓࠋ࡞࠾ࠊ࣎ࣜࢥࢼࢰ࣮ࣝᢞ⩌࡛ ࡣᖜࡢࡳࢆホ౯ᑐ㇟ࡋࡓࠋ
14 2.5. ⤫ィᏛⓗゎᯒ
࣎ࣜࢥࢼࢰ࣮ࣝ࠶ࡿ࠸ࡣࢪࢦ࢟ࢩࣥࡢᢞᚋグ㘓ࡋࡓ standard full-field ERGs ࡢྛィ
್ࡣᖹᆒ್sᶆ‽೫ᕪ㸦SD㸧࡛⾲ࡋࠊᑐᛂࡢ࠶ࡿ t ᳨ᐃࢆ⏝࠸࡚ᢞ๓್㸦፹యᢞᚋࡢ ್㸧ẚ㍑ࡋࡓࠋP<0.05 ࢆࡶࡗ࡚⤫ィᏛⓗ᭷ព࡞ኚุ᩿ࡋࡓࠋ 3. ᐇ㦂⤖ᯝ 3.1. ࣎ࣜࢥࢼࢰ࣮ࣝ 3.1.1. ERG Ἴᙧኚࡢ௦⾲ࢆFig. 5 ♧ࡍࠋࡲࡓࠊゎᯒࡋࡓྛἼᙧᡂศࢆ Tab. 1 ♧ࡍࠋ ྛἼᙧᡂศࡣࠊᢞ⏝㔞㛵㐃ࡋ࡚௨ୗࡢኚࡀㄆࡵࡽࢀࡓࠋRod response ࡛ࡣ b Ἴࡢ 㢧ⴭ࡞ῶᙅ࠶ࡿ࠸ࡣᾘኻࡀㄆࡵࡽࢀࡓࠋᢞ┤ᚋࡢ b Ἴᖜࡣࠊᢞ๓್ẚ㍑ࡋ࡚᭷ព ࡞ప್࡛࠶ࡗࡓ㸦3 mg/kg ᢞ P<0.05ࠊ6 ཬࡧ 12 mg/kg ᢞ P<0.01㸧ࠋCombined rod-cone response ࡣ negative ERG㸦b Ἴᖜࡀ a Ἴᖜࡼࡾࡶᑠࡉ࠸㸧࡛࠶ࡾࠊb ἼཬࡧᚊືᵝᑠἼ
ࡢῶᙅࡀ㢧ⴭࡔࡗࡓࠋᢞ┤ᚋࡢb Ἴᖜཬࡧ b/a ẚࡣࠊᢞ๓್ẚ㍑ࡋ࡚᭷ព࡞ప್࡛
࠶ࡗࡓ㸦12 mg/kg ᢞ P<0.01㸧ࠋa Ἴᖜࡣ᭷ព࡞ኚࡣㄆࡵࡽࢀ࡞ࡗࡓࠋSingle-flash cone response ࡣ㍍ᗘኚࡋࠊa Ἴࢺࣛࣇࡢᖹᆠཬࡧ b Ἴࡢ㍍ᗘ࡞ῶᙅࡀほᐹࡉࢀࡓࠋᢞ
┤ᚋࡢ b Ἴᖜࡣࠊᢞ๓್ẚ㍑ࡋ࡚᭷ព࡞ప್࡛࠶ࡗࡓ㸦6 ཬࡧ 12 mg/kg ᢞ P<0.01㸧ࠋaἼᖜࡣ᭷ព࡞ኚࡣㄆࡵࡽࢀ࡞ࡗࡓࠋྠᵝ 30 Hz flicker ࡶ㍍ᗘኚࡋࠊ b Ἴࡢ㍍ᗘ࡞ῶᙅࡀほᐹࡉࢀࡓࠋᢞ┤ᚋࡢ b Ἴᖜࡣࠊᢞ๓್ẚ㍑ࡋ࡚᭷ព࡞ప್࡛ ࠶ࡗࡓ㸦6 mg/kg ᢞ P<0.05ࠊ12 mg/kg ᢞ P<0.01㸧ࠋᢞ┤ᚋㄆࡵࡽࢀࡓࡍ࡚ࡢ ኚࡣࠊྛᢞࡢ 24 㛫ᚋࡣᅇࡋࡓࠋࡲࡓୖグࡢ ERG ࡢኚࠊ᫂ࡽ࡞㞤 㞝ᕪࡣほᐹࡉࢀ࡞ࡗࡓࠋ
3.1.2. ║ᗏ᳨ᰝ ࠸ࡎࢀࡢほᐹ࣏ࣥࢺ࠾࠸࡚ࡶ᫂ࡽ࡞␗ᖖࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ 3.2. ࢪࢦ࢟ࢩࣥ 3.2.1. ERG Ἴᙧኚࡢ௦⾲ࢆFig. 6 ♧ࡍࠋࡲࡓࠊゎᯒࡋࡓྛἼᙧᡂศࢆ Tab. 2 ♧ࡍࠋ Rod response ࡛ࡣᛂ⟅ࡢ㐜ᘏཬࡧቑᖜࡀㄆࡵࡽࢀࠊᢞᚋࡢ b Ἴᢞ๓್ẚ㍑ࡋ࡚᭷
ព࡞₯ᘏ㛗ཬࡧᖜቑຍ㸦ࡶp<0.01㸧ࡀ᳨ฟࡉࢀࡓࠋCombined rod-cone response ࡛
ࡣᛂ⟅ࡢ㐜ᘏࡀㄆࡵࡽࢀࠊᢞᚋࡢaἼཬࡧ bἼᢞ๓್ẚ㍑ࡋ࡚᭷ព࡞₯ࡢᘏ㛗㸦
ࡶ p<0.01㸧ࡀ᳨ฟࡉࢀࡓࠋᖜࡣ a Ἴ࠶ࡿ࠸ࡣ b Ἴࡢ࠸ࡎࢀࡶⴭኚࡣほᐹࡉࢀ࡞
ࡗࡓࠋSingle-flash cone response ཬࡧ 30 Hz flicker ࡛ࡣࠊࡶᛂ⟅ࡢ㐜ᘏࡀㄆࡵࡽࢀࡓࠋ୧
ᛂ⟅ࡢᢞᚋࡢ b Ἴ₯ࡣࠊᢞ๓್ẚ㍑ࡋ࡚᭷ពᘏ㛗ࡋࡓ㸦ࡶ p<0.01㸧ࠋࡲࡓࠊ
⤫ィᏛⓗ᭷ពᕪࡣ᳨ฟࡉࢀ࡞ࡗࡓࡀࠊᢞᚋࡢsingle-flash cone response ཬࡧ 30 Hz flicker
ࡢb Ἴᖜࡣ 5/6 ࡛ᢞ๓ࡼࡾࡶప್ࢆ♧ࡋࡓࠋୖグࡢࡍ࡚ࡢኚࡣᢞ 24 㛫ᚋࡀ
᭱ࡶ㢧ⴭ࡛࠶ࡗࡓࠋSingle-flash cone response ࡢ b Ἴㄆࡵࡽࢀࡓ₯ᘏ㛗ࡣᢞ 24 㛫ᚋ
௨㝆ᅇഴྥࢆ♧ࡋࡓࡶࡢࡢࠊᢞ28 ᪥ᚋ࡛ࡢ b Ἴ₯ࡣᢞ๓್ẚ㍑ࡋ࡚᭷ព࡞㧗
್㸦p<0.05㸧࡛࠶ࡗࡓࠋࡑࢀ௨እࡢࡍ࡚ࡢኚࡣࠊᢞ 28 ᪥ᚋࡲ࡛ᢞ๓ྠ⛬ᗘࡲ
࡛ᅇࡋࡓࠋࡲࡓࠊୖグࡢERG ࡢኚࠊ᫂ࡽ࡞㞤㞝ᕪࡣほᐹࡉࢀ࡞ࡗࡓࠋ
3.2.2. ║ᗏ᳨ᰝ
16 ᰁࡢ⒪⸆ࡋ࡚⏝ࡉࢀ࡚࠸ࡿࠋ࣎ࣜࢥࢼࢰ࣮ࣝࡣࣄࢺᑐࡋ࡚ᢞᚋ⩈᫂ࠊ㟝どࠊ Ⰽぬࡢኚ➼ࡢどぬ㞀ᐖࢆ୍㐣ᛶㄏⓎࡍࡿࡇࡀሗ࿌ࡉࢀ࠾ࡾࠊ⮫ᗋヨ㦂࡚ホ౯ࡉࢀ ࡓERG ࡢ⤖ᯝࡽࡣ࣎ࣜࢥࢼࢰ࣮ࣝࡀ⥙⭷ᶵ⬟ࠊ࡞࡛ࡶᮞయ⤒㊰ࡢᶵ⬟㢧ⴭ࡞ᙳ㡪ࢆ ཬࡰࡍࡇࡀ♧၀ࡉࢀࡿ(12)ࠋ ᮏᐇ㦂ࡢ࢝ࢽࢡࢨ࡛ࣝࡣࠊ࣎ࣜࢥࢼࢰ࣮ࣝࡢᢞᚋᮞయ⣔ᛂ⟅ࡣᾘኻࡋ࡚ΰྜᛂ⟅ ࡣᚊືᵝᑠἼࡢῶᙅࢆక࠺ negative ERG ࢆ♧ࡋࠊᮞయ⤒㊰ࡢ㢧ⴭ࡞ᙳ㡪ࡀㄆࡵࡽࢀࡓࠋ ୍᪉ࠊ㗹య⣔ᛂ⟅ࡣῶᙅࡀㄆࡵࡽࢀࡓࡶࡢࡢ㍍ᗘ࡛࠶ࡾࠊ㗹య⤒㊰ࡢᙳ㡪ࡣᮞయ⤒㊰ ẚ㍑ ࡍࡿ㍍ᗘ࡛࠶ࡗ ࡓࠋ௨ୖࡼࡾࠊᮏ ᐇ㦂᳨࡚ฟࡉࢀ ࡓ࣎ࣜࢥࢼࢰ࣮ࣝ ࡼࡿ standard full-field ERGs ࡢኚࡣࠊࣄࢺ࠾ࡅࡿኚ୍⮴ࡋࡓࡶࡢ⪃࠼ࡓࠋ
ࡉࡽࠊᮏᐇ㦂࡚グ㘓ࡋࡓstandard full-field ERGs ࡢࡍ࡚ࡢᛂ⟅࡛ࡋ࡚ᴟ⣽⬊
⏤᮶ࡍࡿb Ἴࡢῶᙅࡀㄆࡵࡽࢀࡓ୍᪉ࠊࡋ࡚ど⣽⬊⏤᮶ࡍࡿ a Ἴࡣ࠸ࡎࢀࡢᛂ⟅ ࡶ᫂ࡽ࡞ኚࡣほᐹࡉࢀ࡞ࡗࡓࠋࡇࡢ⤖ᯝࡽࠊ࣎ࣜࢥࢼࢰ࣮ࣝࡢᢞࢆཷࡅࡓ࢝ ࢽࢡࢨ࡛ࣝࡣࠊど⣽⬊ࡢᶵ⬟ࡣẚ㍑ⓗ Ꮡࡉࢀ࡚࠸ࡿ୍᪉ࠊᴟ⣽⬊ࡢᶵ⬟ࡣ㢧ⴭ㞀 ᐖࡉࢀࡿࡇࡀᙉࡃ♧၀ࡉࢀࡓࠋ ࣄࢺ⮫ᗋ࡚࠺ࡗ⾑ᛶᚰཪࡣ㢖⬦ࡢ⒪⸆ࡋ࡚⏝ࡉࢀ࡚࠸ࡿࢪࢦ࢟ࢩࣥࡣࠊ୰ ẘᝈ⪅どຊపୗࠊ㟝どࠊ㯤ど➼ࡢどぬ␗ᖖࢆྍ㏫ⓗㄏⓎࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ(8, 39, 49, 54, 74)ࠋሗ࿌(49, 74)࠾࠸࡚ࢪࢦ࢟ࢩࣥ୰ẘᝈ⪅࡛ホ౯ࡉࢀࡓERG ࡢ⤖ᯝࡽࡣࠊ㗹య⤒ ㊰ࡢྍ㏫ⓗ࡞ᶵ⬟㞀ᐖࡀᙉࡃ♧၀ࡉࢀࡿ୍᪉ࠊᮞయ⤒㊰ࡢᛂࡣࡴࡋࢁቑഴྥࢆ♧ࡋ࡚ ࠸ࡓ᥎ᐹࡉࢀࡿࠋ ᮏᐇ㦂ࡢ࢝ࢽࢡࢨ࡛ࣝࡣࠊ㗹య⣔ᛂ⟅ࡢ₯ᘏ㛗ཬࡧᖜࡢపୗഴྥࡀㄆࡵࡽࢀࠊ㗹 య⤒㊰ࡢ㞀ᐖࢆᙉࡃ♧၀ࡍࡿ⤖ᯝࡀᚓࡽࢀࡓࠋ୍᪉ࠊᮞయ⣔ᛂ⟅ࡣቑᖜࢆ♧ࡋࠊᮞయ⤒㊰ ࡢᛂࡀቑࡋ࡚࠸ࡓࡇࡀ♧၀ࡉࢀࡓࠋࡲࡓࡇࢀࡽࡢERG ࡢኚࡣྍ㏫ⓗ࡛࠶ࡗࡓࠋ௨
ୖࡼࡾࠊᮏᐇ㦂ࡢ࢝ࢽࢡࢨ᳨࡚ࣝฟࡉࢀࡓࢪࢦ࢟ࢩࣥࡼࡿstandard full-field ERGs ࡢ
5. ᑠᣓ
ࣄࢺ࡛⸆ㄏⓎᛶࡢERG ࡢኚࢆ♧ࡍࡇࡀ▱ࡽࢀ࡚࠸ࡿ࣎ࣜࢥࢼࢰ࣮ࣝཬࡧࢪࢦ࢟ࢩ
ࣥࢆࠊࡑࢀࡒࢀ࢝ࢽࢡࢨࣝᢞࡋ࡚standard full-field ERGs ࢆグ㘓ࡋࡓࠋࡑࡢ⤖ᯝࠊࡇ
ࢀࡽࡢ⸆ࢆᢞࡉࢀࡓ࢝ࢽࢡࢨ࡛ࣝࡣࠊࡑࢀࡒࢀᮞయ⤒㊰ཬࡧ㗹య⤒㊰㑅ᢥⓗࡘ ྍ㏫ⓗ࡞⥙⭷ᶵ⬟㞀ᐖࢆ♧၀ࡍࡿኚࡀ᳨ฟࡉࢀࡓࠋ๓㏙ࡢኚࡣࣄࢺ࡛ࡢሗ࿌࡛♧ࡉࢀ
ࡓ⤖ᯝ୍⮴ࡋ࡚࠸ࡓࡇࡽࠊ࢝ࢽࢡࢨ࡛ࣝࡢstandard full-field ERGs グ㘓ࡼࡗ࡚ࠊ
࣎ࣜࢥࢼࢰ࣮ࣝࡢᮞయ⤒㊰ࡢ㢧ⴭ࡞ᙳ㡪ࠊཬࡧࢪࢦ࢟ࢩࣥࡢ㗹య⤒㊰ࡢ㢧ⴭ࡞ᙳ㡪ࢆ ᳨ฟྍ⬟࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋ
18
FIGURE 5. Typical waveforms of the standard full-field ERGs in a voriconazole-treated
cynomolgus monkey. Voriconazole was administered intravenously with increasing doses of 0 (vehicle), 3, 6 and 12 mg/kg at intervals of one week or more, and the standard full-field ERGs were obtained as described in the text. Arrowheads indicate onset of the light flashes. The responses recorded 24 hours after each dosing (gray trace) are superimposed on those recorded immediately after each dosing (black trace). Each trace represents an average of three responses.
FIGURE 6. Typical waveforms of the standard full-field ERGs in a digoxin-treated cynomolgus
monkey. Digoxin was administered intravenously at a dose of 0.1 mg/kg, and the standard full-field ERGs were obtained as described in the text. Arrowheads indicate onset of the light flashes. The responses recorded before dosing (gray trace) are superimposed on those recorded after dosing (black trace). Each trace represents an average of three responses.
20
TABLE 1. Effects of Voriconazole on the Standard Full-Field ERGs in Cynomolgus Monkeys
Data are expressed as the mean ± SD of six animals.
All the animals were treated with low to high doses at intervals of one week or more. * Physiological saline was administered as a vehicle control.
͊ p<0.05, ‡ p<0.01; significantly different in comparison with the vehicle-control value by the paired t-test
Rod Response Amplitude (μV)
b-wave 43.9 㼼9.97 34.2 㼼13.80 44.8 㼼13.93 16.5 㼼6.47 47.7 㼼 8.23 9.7 㼼5.73 40.8 㼼12.03 Combined Rod-Cone Response
Amplitude (μV)
a-wave 52.8 㼼9.10 49.0 㼼8.97 51.0 㼼7.50 45.4 㼼8.77 54.8 㼼 6.53 46.1 㼼12.40 63.7 㼼8.50 b-wave 123.5 㼼24.93 121.9 㼼22.94 126.1 㼼21.81 89.9 㼼14.79 132.5 㼼 19.17 60.5 㼼13.88 135.7 㼼21.41 Single-Flash Cone Response
Amplitude (μV) a-wave 12.7 㼼1.81 12.9 㼼2.36 12.7 㼼1.21 11.1 㼼2.34 13.5 㼼 2.22 11.0 㼼2.67 14.6 㼼2.31 b-wave 53.8 㼼6.63 47.7 㼼7.17 55.3 㼼5.42 41.3 㼼5.77 60.3 㼼 11.23 33.0 㼼5.11 64.2 㼼12.23 30 Hz Flicker Amplitude (μV) b-wave 52.4 㼼6.62 48.1 㼼8.04 55.3 㼼3.80 45.6 㼼6.08 59.6 㼼 12.32 35.3 㼼5.09 58.1 㼼11.89 Immediately After 24 h After Dose (mg/kg) 0* 3 6 12 Immediately After Immediately After 24 h After Immediately After 24 h After ‡ ‡ † † ‡ ‡ ‡ ‡
TABLE 2. Effects of Digoxin on the Standard Full-Field ERGs in Cynomolgus Monkeys
Data are expressed as the mean ± SD of six animals. 1) Recorded 24 hours after dosing of physiological saline.
͊ p<0.05, ‡ p<0.01; significantly different in comparison with the vehicle-control value by the paired t-test Rod Response Implicit Time (ms) b-wave 83.5 㼼 2.82 87.0 㼼 2.27 91.3 㼼 4.49 89.5 㼼 5.42 84.2 㼼 1.95 84.8 㼼 2.52 Amplitude (μV) b-wave 49.0 㼼 6.60 58.9 㼼 16.58 72.3 㼼 12.32 59.8 㼼 20.06 47.5 㼼 14.40 51.3 㼼 9.48 Combined Rod-Cone Response
Implicit Time (ms) a-wave 17.3 㼼 0.48 17.4 㼼 0.30 18.4 㼼 0.64 18.0 㼼 0.69 17.8 㼼 0.78 17.3 㼼 0.44 b-wave 38.0 㼼 1.81 39.4 㼼 1.66 42.9 㼼 2.56 40.9 㼼 3.29 38.7 㼼 1.81 38.2 㼼 1.41 Amplitude (μV) a-wave 58.4 㼼 8.44 58.6 㼼 6.89 57.5 㼼 8.09 57.9 㼼 9.16 66.8 㼼 15.83 65.0 㼼 9.59 b-wave 141.5 㼼 16.36 138.6 㼼 19.59 148.2 㼼 11.19 168.6 㼼 23.35 158.0 㼼 31.08 159.0 㼼 23.94 Single-Flash Cone Response
Implicit Time (ms) a-wave 11.6 㼼 0.43 11.8 㼼 1.08 12.4 㼼 1.09 12.6 㼼 0.88 12.0 㼼 0.42 11.9 㼼 0.34 b-wave 23.9 㼼 0.71 24.7 㼼 1.00 26.2 㼼 1.00 26.1 㼼 1.11 25.1 㼼 0.61 24.6 㼼 0.69 Amplitude (μV) a-wave 14.4 㼼 2.64 12.3 㼼 1.10 13.7 㼼 2.45 14.7 㼼 4.09 14.8 㼼 1.70 13.5 㼼 2.81 b-wave 57.7 㼼 10.90 53.0 㼼 7.41 47.9 㼼 7.85 53.7 㼼 14.71 60.5 㼼 7.31 59.5 㼼 10.69 30 Hz Flicker Implicit Time (ms) b-wave 25.7 㼼 0.67 26.3 㼼 0.69 27.1 㼼 0.69 26.9 㼼 0.77 26.7 㼼 0.88 26.2 㼼 0.59 Amplitude (μV) b-wave 57.8 㼼 6.77 54.5 㼼 9.51 47.8 㼼 9.96 54.5 㼼 15.36 61.1 㼼 8.57 61.5 㼼 6.55 Times After Dosing
Baseline 1) Immediately 24 hours 7 days 14 days 28 days
‡ † ‡ ‡ ‡ † † ‡ ‡ † ‡ ‡ † ‡ ‡ † ‡ ‡ †
22
➨
2 ❶ a-wave analysis㸸୍ḟࢽ࣮ࣗࣟࣥ㸦ど⣽⬊㸧ࡢホ౯
1. ᗎㄽ ⥙⭷ࡢ୍ḟࢽ࣮࡛ࣗࣟࣥ࠶ࡿど⣽⬊ࡣࠊࡑࡢእ⠇Ꮡᅾࡍࡿど≀㉁࡚ගᏊࢆ྾ࡋࠊ ⤖ᯝࡋ࡚⮬ࡽࡢ⭷㟁ࢆኚࡉࡏࡿගཷᐜయ࡛࠶ࡿࠋど≀㉁ࡼࡿගᏊࡢ྾ࡽ⭷㟁 ࡢኚ⮳ࡿࡲ࡛ࡢ㐣⛬ࡣගሗኚ㸦Fig. 7㸧ࡤࢀࠊࡇࢀࡲ࡛ከࡃࡢ◊✲⪅ࡼࡗ ࡚ㄪࡽࢀ࡚ࡁࡓࠋࡇࡢගሗኚࡢ⤖ᯝ࡛࠶ࡿど⣽⬊ࡢ⭷㟁ࡢኚࡣࠊERG ࡛ࡣග่ ⃭ࡢᚋࡢ᭱ึࡢ㝜ᛶἼ࡛࠶ࡿa Ἴ㸦Fig. 4㸧᭱ࡶᫎࡉࢀࡿࡇࡀࡼࡃ▱ࡽࢀࡿࠋࡑࡢࡓ ࡵࠊ୍⯡ⓗERG ࡢ a Ἴࡣど⣽⬊ᶵ⬟ࡢᣦᶆࡉࢀࡿࠋ➨1 ❶࡛ࡣࠊ࢝ࢽࢡࢨ࡛ࣝࡢ standard full-field ERGs グ㘓ࡼࡗ࡚⸆ࡼࡿᮞయ⤒㊰
࠶ࡿ࠸ࡣ㗹య⤒㊰ࡢ㑅ᢥⓗ࡞స⏝ࡀ᳨ฟྍ⬟࡛࠶ࡿࡇࢆ♧ࡋࡓࠋࡋࡋ ࠊstandard
full-field ERGs ࡢ⤖ᯝࡀど⣽⬊ࡢస⏝ࢆ♧၀ࡍࡿࡶࡢ࡛࠶ࡗࡓ㝿ࠊࡑࡢస⏝ࢆࡼࡾヲ⣽ ㄪࡿࡓࡵࡣࠊど⣽⬊ࡢᶵ⬟ホ౯↔Ⅼࢆᙜ࡚ࡓูࡢᡭἲࡀᚲせ࡞ࡿࠋࡍ࡞ࢃࡕࠊ standard full-field ERGs ど⣽⬊ᮞయࡢᶵ⬟ࢆ┤᥋ᫎࡍࡿἼᙧᡂศࡣྵࡲࢀࡎࠊࡲࡓࠊ୍
⯡ⓗど⣽⬊㗹యࡢᶵ⬟ࡢᣦᶆࡉࢀࡿstandard full-field ERGs ࠾ࡅࡿ᫂㡰ᛂୗ ERG ࡢ a
Ἴࡣࠊ㗹యࡢᛂຍ࠼࡚㗹యOFF ⤒㊰㸦OFF ᆺᴟ⣽⬊ཬࡧỈᖹ⣽⬊㸧ࡢᛂࡶከศ ྵࡲࢀࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿࡽ࡛࠶ࡿ(5, 23)ࠋ ୍᪉ࠊHood ࡽ(24, 27)ࡣࣄࢺࡢᝈ⪅࡛ど⣽⬊ᮞయࡢᶵ⬟ࢆṇ☜ࡘヲ⣽ᢕᥱࡍࡿࡓࡵࠊERG ࡢa Ἴࡢึᮇ㒊ศࢆᮞయࡢගሗኚ⏤᮶ࡍࡿᩘᘧࣔࢹࣝࣇࢵࢸࣥࢢࡋࠊᚓࡽࢀࡓ ࣔࢹࣝࣃ࣓࣮ࣛࢱࡽගሗኚ㐣⛬ࢆྵࡵ࡚ᮞయᶵ⬟ࢆホ౯ࡍࡿᡭἲࢆ⪃ࡋࡓ㸦a-wave analysis㸧ࠋࡉࡽ㗹యࡘ࠸࡚ࡶྠᵝࠊHood ࡽࡼࡗ࡚ୖグࡢᮞయᶵ⬟ホ౯ἲ㢮ఝࡢ ᪉ἲࡀ☜❧ࡉࢀ࡚࠸ࡿ(26)ࠋࡇࢀࡽࡢ᪉ἲࢆ࢝ࢽࢡࢨ࡛ࣝࡢᏳᛶヨ㦂ᛂ⏝ࡍࡿࡇ࡛ࠊ ⸆ࡼࡿど⣽⬊ᶵ⬟ࡢస⏝ࢆヲ⣽ホ౯࡛ࡁࡿࡼ࠺࡞ࡿᮇᚅࡉࢀࡿࠋ
㉳ཬࡧ⫵ື⬦ᛶ⫵㧗⾑ᅽࡢ⒪⸆ࡋ࡚⏝࠸ࡽࢀ࡚࠸ࡿࢩࣝࢹࢼࣇࣝࡣࠊᝈ⪅
୍㐣ᛶࡢどぬ㞀ᐖࢆㄏⓎࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ(33)ࠋࢩࣝࢹࢼࣇࣝࡣ 6 ᆺ࣍ࢫ࣍ࢪ࢚ࢫ
ࢸ࣮ࣛࢮ㸦PDE type 6: PDE6㸧ᑐࡍࡿ㜼ᐖస⏝ࡶ᭷ࡋ(77)ࠊࡇࡢPDE6 ࡣど⣽⬊እ⠇࠾ࡅ
ࡿගሗኚࡢ㔜せ࡞㓝⣲ࡢ୍ࡘ࡛࠶ࡿࠋࡑࡢࡓࡵࠊୖグࡢどぬ㞀ᐖࡣࢩࣝࢹࢼࣇࣝ ࡼࡿど⣽⬊ࡢ PDE6 㜼ᐖ㛵㐃ࡋࡓࡶࡢ⪃࠼ࡽࢀ࡚࠸ࡿ(33)ࠋࡲࡓࠊࣄࢺ࡛ྍ㏫ⓗ࡞どぬ 㞀 ᐖ ࢆ ㄏ Ⓨ ࡍ ࡿ ࡇ ࡀ ▱ ࡽ ࢀ ࡿ ࢪ ࢦ ࢟ ࢩ ࣥ ࢆ ྵ ࡴ ࢪ ࢠ ࢱ ࣜ ࢫ ࡣ ࠊ ど ⣽ ⬊ ෆ ⠇ ࡢ Na+-K+ATPase ࡢάᛶࢆ㜼ᐖࡍࡿ(4)ࠋࡇࡢど⣽⬊ࡢNa+-K+ATPase ࡣࠊ⣽⬊ෆ K+ࢆ䬦ࡳධࢀ ࡿࡇ࡛ど⣽⬊ࡢගᛂ⟅㔜せ࡞ᙺࢆᯝࡓࡍࡇࡽࠊୖグࡢどぬ㞀ᐖࡣࢪࢦ࢟ࢩࣥ ࡼࡿど⣽⬊ࡢNa+-K+ATPase 㜼ᐖ㛵㐃ࡋࡓࡶࡢ⪃࠼ࡽࢀ࡚࠸ࡿ(39)ࠋ ࡑࡇ࡛ᮏ◊✲࡛ࡣࠊ㠀⮫ᗋᏳᛶヨ㦂୍⯡ⓗ⏝࠸ࡽࢀࡿື≀✀࡛࠶ࡿ࢝ࢽࢡࢨࣝ
ࢆ⏝࠸࡚ࠊHood ࡽࡢ᪉ἲ(28)ࢆཧ⪃a-wave analysis ࡢホ౯᮲௳ࢆタᐃࡋࡓࠋࡲࡓࠊྠ a-wave
analysis ࡚ࠊ๓㏙ࡢࢩࣝࢹࢼࣇࣝཬࡧࢪࢦ࢟ࢩࣥࡼࡿど⣽⬊ࡢస⏝ࢆᫎࡋࡓኚ ࡀ᳨ฟྍ⬟᳨ドࡋࡓࠋ 2. ᐇ㦂ᮦᩱཬࡧᐇ㦂᪉ἲ ᮏᐇ㦂ࡣࠊ➨୍୕ඹᰴᘧ♫ࡢື≀ᐇ㦂ጤဨࡢᢎㄆ㸦ື≀ᐇ㦂ᢎㄆ␒ྕ㸸A1200207 ཬ ࡧC800264㸧ࢆᚓ࡚ᐇࡋࡓࠋ 2. ື≀ 2.1. ື≀ ୍⯡≧ែཬࡧ║ᗏ␗ᖖࡢ࡞࠸㞤㞝ྛ 12 ࠊィ 24 ࡢ࣋ࢺࢼ࣒⏘࠶ࡿ࠸ࡣࣥࢻࢿࢩ ⏘ࡢ࢝ࢽࢡࢨࣝ㸦3㸫5 ṓ㱋ࠊయ㔜 2.6㸫5.1 kg㸧ࢆ⏝࠸ࡓࠋணഛ᳨ウࢆ⾜ࡗࡓ⤖ᯝࠊERG
24 㱋ࠊయ㔜2.5㸫3.4 kg㸧ࠊࢩࣝࢹࢼࣇࣝ 1 mg/kg ⩌㸦4 ṓ㱋ࠊయ㔜 2.6㸫3.0 kg㸧ࠊࢩࣝࢹࢼࣇ ࣝ3 mg/kg ⩌㸦4㸫5 ṓ㱋ࠊయ㔜 2.6㸫3.4 kg㸧ࠊཬࡧࢩࣝࢹࢼࣇࣝ 10 mg/kg ⩌㸦3㸫4 ṓ 㱋ࠊయ㔜 2.6㸫3.2 kg㸧ࡑࢀࡒࢀࡾศࡅࡓࠋࢪࢦ࢟ࢩࣥᢞᐇ㦂࡛ࡣࠊྛ 4 ࢆ፹యᑐ ↷⩌㸦4㸫5 ṓ㱋ࠊయ㔜 2.6㸫4.6 kg㸧ཬࡧࢪࢦ࢟ࢩࣥ⩌㸦3㸫5 ṓ㱋ࠊయ㔜 2.6㸫5.1 kg㸧ࡑ ࢀࡒࢀࡾศࡅࡓࠋື≀ࡣ1 ❶ྠᵝ㣫⫱ࡋࡓࠋ 2.2. ᢞ ど⣽⬊ࡢᶵ⬟ྍ㏫ⓗ࡞ᙳ㡪ࢆཬࡰࡍࡇࡀ▱ࡽࢀ࡚࠸ࡿ⸆ࡋ࡚ࠊࢩࣝࢹࢼࣇࣝ ཬࡧࢪࢦ࢟ࢩࣥࢆ㑅ᢥࡋࡓࠋ
ࢩࣝࢹࢼࣇࣝ㸦Sildenafil citrate salt; Sigma-Aldrich, St. Louis, USA㸧ࡣ⏕⌮㣗ሷᾮ⁐ゎ
ࡉࡏࡓᚋࠊᏍᚄ0.2 μm ࡢࢩࣜࣥࢪࣇࣝࢱ࣮㸦Minisart; Sartorius stedim biotech, Gottingen,
Germany㸧ࢆ⏝࠸࡚ࢁ㐣⁛⳦ࡋ࡚ᢞᾮࢆㄪ〇ࡋࡓࠋᢞᾮࡣࠊྛ 4 ࡽ࡞ࡿ፹యᑐ↷⩌ࠊ
ࢩࣝࢹࢼࣇࣝ1ࠊ3 ཬࡧ 10 mg/kg ⩌ࠊࡑࢀࡒࢀ 0ࠊ1ࠊ3 ཬࡧ 10 mg/kg ࡢ⏝㔞࡚༢ᅇ
㟼⬦ෆᢞ㸦0.2 mL/kg/min10 min㸧ࡋࡓࠋᢞ⏝㔞ࡣࠊࣄࢺࡢ⮫ᗋྠ⛬ᗘࡢ᭚㟢ࢆぢ㎸
ࡵࡿ⏝㔞ࡋ࡚1 mg/kg ࢆタᐃࡋࡓࠋᢞࡣࠊࢩࣜࣥࢪ࣏ࣥࣉ㸦22; Harvard Apparatus, Inc.㸧
ࢆ⏝࠸࡚అᅾ㟼⬦ࡽ⾜ࡗࡓࠋ ࢪࢦ࢟ࢩࣥ⩌ࡢ4 ࡣࠊ➨ 1 ❶ྠᵝࢪࢦ࢟ࢩࣥࢆ 0.1 mg/kg ࡢ⏝㔞࡚༢ᅇ㟼⬦ෆ ᢞࡋࠊ፹యᑐ↷⩌ࡢ4 ࡣ⏕⌮㣗ሷᾮࢆྠᵝᢞࡋࡓࠋ 2.3. ERG 2.3.1. ๓ฎ⨨ ື≀ࡣࠊ➨1 ❶ྠᵝሷ㓟ࢣࢱ࣑ࣥ㸦ᑟධ㸸10 mg/kgࠊ⥔ᣢ㸸5-10 mg/kg/h㸧ཬࡧ࢟ࢩ ࣛࢪࣥሷ㓟ሷ0.6 mg/kg ࡢ➽⫗ෆᢞࡼࡾ㯞㓉ࡋࡓࠋ▖Ꮝࡣࢺࣟࣆ࣑࢝ࢻཬࡧࣇ࢙ࢽࣞࣇ ࣜࣥሷ㓟ሷࡢⅬ║ࡼࡾᩓ▖ࡉࡏࡓࠋゅ⭷ࡣሷ㓟࢜࢟ࢩࣈࣉࣟ࢝ࣥࡢⅬ║ࡼࡾ㯞㓉ࡋ ࡓᚋࠊࣄࢻࣟ࢟ࢩ࢚ࢳࣝࢭ࣮ࣝࣟࢫࡢⅬ║ࡼࡾಖㆤࡋࡓࠋ
2.3.2. ග่⃭
ERG ࡢㄏⓎࡣⓎගࢲ࣮࢜ࢻ㸦light-emitting diode: LED㸧ཬࡧ࢟ࢭࣀࣥࣛࣥࣉࡢ୧ග※
ࢆෆⶶࡋࡓど㔝่⃭⨨㸦BigShotGanzfeld; LKC technologies, Inc.㸧ࢆ⏝ࡋࡓࠋᮏᐇ㦂࡛ ࡣࠊ࢟ࢭࣀࣥࣛࣥࣉග※ࡢࡳࢆ⏝ࡋࡓࠋ࢟ࢭࣀࣥࣛࣥࣉࡼࡿග่⃭ࡢ᭱ᙉᗘ㸦ᐇ
್㸧ࡣ826.7 cd∙s/m2࡛࠶ࡾࠊ่⃭᮲௳ࡣ่⃭ไᚚ⏝PC ࣥࢫࢺ࣮ࣝࡉࢀࡓࢯࣇࢺ࢚࢘
㸦Ganzfeld control panel; LKC technologies, Inc.㸧࡚ไᚚࡋࡓࠋග่⃭ᙉᗘࡢ༢ࡣࠊHood ࡽࡢሗ࿌(24, 26)୍⮴ࡉࡏ࡚⥙⭷↷ᗘ㸦troland㸸td㸧࡚⾲ࡋࠊᬯᡤど㍤ᗘࡣࠕscot tdࠖࠊ᫂ᡤ ど㍤ᗘࡣࠕphot tdࠖࡑࢀࡒࢀ⾲グࡋࡓࠋ
2.3.3. グ㘓ཬࡧゎᯒ
Hood ࡽࡢሗ࿌(24, 26, 27)ࢆཧ⪃ࠊa-wave analysis㸦ERG ࡢグ㘓ࠊῶ⟬ࠊཬࡧࣇࢵࢸࣥ
ࢢ㸧ࢆ௨ୗࡢࡼ࠺ᐇࡋࡓࠋ
グ㘓㟁ᴟࡋ࡚ᴟᆺࢥࣥࢱࢡࢺࣞࣥࢬ㟁ᴟ㸦H6515NFC㸹᭷㝈♫࣓࣮ࣚ㸧ࢆᕥ║ࠊ ᥋ᆅ㟁ᴟࡋ࡚㔪㟁ᴟ㸦TN208-016㸹ᰴᘧ♫ࣘࢽ࣮ࢡ࣓ࢹ࢝ࣝ㸧ࢆ㢌㡬㒊ࡢ⓶ୗࡑࢀ ࡒࢀタ⨨ࡋࡓࠋ40 ศ௨ୖࡢᬯ㡰ᛂᚋࠊ3.7ࠊ4.0ࠊཬࡧ 4.3 log scot td-s ࡢᙉᗘࡢග่⃭ᑐࡍ
ࡿERG ࡛࠶ࡿΰྜᛂ⟅㸦combined rod-cone response㸧ࢆ㡰ḟㄏⓎࡋࡓࠋ࡞࠾ࠊᮞయࡢᬯ㡰
ᛂࢆ⥔ᣢࡍࡿࡓࡵࠊග่⃭ࡢ㛫㝸ࡣ30 ⛊㛫௨ୖタࡅࡓࠋࡑࡢᚋࠊpaired-flash technique ࢆ⏝
࠸࡚ΰྜᛂ⟅ྠ୍ᙉᗘࡢ୍㐃ࡢග่⃭㸦3.4ࠊ3.7ࠊཬࡧ 4.0 log phot td-s㸧࡚㗹య⣔ᛂ⟅
㸦cone response㸧ࢆㄏⓎࡋࡓࠋpaired-flash technique ࡼࡿ㗹య⣔ᛂ⟅ࡢグ㘓ࡣ Friedburg ࡽ(16)
ཬࡧJeffrey ࡽ(30)ࡢሗ࿌ࢆཧ⪃ࠊᬯ㡰ᛂୗ࡚4.0 log scot td-s ࡢᙉᗘࡢ conditioning flash
26 ࢩࣝࢹࢼࣇࣝᢞᐇ㦂࡛ࡣᢞ๓ࠊᢞ┤ᚋཬࡧᢞ24 㛫ᚋࠊࢪࢦ࢟ࢩࣥᢞᐇ㦂 ࡛ࡣᢞ๓ࠊᢞ24 㛫ཬࡧ 1 ࣧ᭶ᚋࡑࢀࡒࢀグ㘓ࡋࡓࠋ ࡍ࡚ࡢᛂ⟅ࡣ 0.5㸫1000 Hz ࡢᖏᇦࣇࣝࢱ࣮ࢆ㏻ࡋ࡚ቑᖜࡋࡓᚋࠊㄏⓎ㟁᳨ᰝ ⨨㸦MEB-9104㸹᪥ᮏග㟁ᰴᘧ♫㸧グ㘓ࡋࡓࠋྠ୍ᙉᗘࡢග่⃭ᑐࡍࡿᛂ⟅ࢆࠊΰྜ ᛂ⟅࡛ࡣ 3 Ἴᙧࠊ㗹య⣔ᛂ⟅࡛ࡣ 6 Ἴᙧグ㘓ࡋࠊࡑࢀࡽࡢຍ⟬ᖹᆒἼᙧࢆ⟬ฟࡋࡓࠋ㗹య ⣔ᛂ⟅ࢆྠ୍ᙉᗘࡢග่⃭࡚ㄏⓎࡉࢀࡓΰྜᛂ⟅ࡽῶ⟬ࡋࠊᮞయ⣔ᛂ⟅㸦rod-isolated
response㸧ࢆồࡵࡓࠋRod-isolated response ࡢ⟬ฟࡢᴫせࢆ Fig. 9 ♧ࡍࠋ
ᮞయ⣔ᛂ⟅ࡢa Ἴࡢึᮇ㒊ศ㸦Rod a-wave㸧ࠊLamb ཬࡧ Pugh ࡢᮞయගሗఏ㐩ࣔࢹࣝ
(32)ࡢHood ཬࡧ Birch ࡼࡿᨵኚࣔࢹࣝ(24)࡛࠶ࡿᘧ (1)ࢆࣇࢵࢸࣥࢢࡋࡓࠋࡘࡲࡾࠊᐇ Ἴᙧ㸦Rod a-wave㸧ᘧ (1)࡛⾲ࡉࢀࡿ᥎ᐃἼᙧࡢṧᕪࡀ᭱ᑡ࡞ࡗࡓ㝿ࡢࣔࢹࣝࣃ ࣓࣮ࣛࢱ㸦ᚋ㏙㸧ࢆồࡵࡓࠋ max 2
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(t > td) (1)R ࡣᖜ㸦μV㸧ࠊI ࡣග่⃭ᙉᗘ㸦log scot td-s㸧ࠊS ࡣឤᗘ㸦s-2(td-s)-1㸧ࠊRmaxࡣ᭱ᖜ㸦μV㸧ࠊ
t ࡣග่⃭ᚋ㛫㸦ms㸧ࠊtdࡣග่⃭ᚋࡢ▷㛫ࡢ㐜ࢀ㸦ms㸧ࢆࡑࢀࡒࢀ⾲ࡍࠋtdࡣᖖ࡞࢝
ࢽࢡࢨࣝ20 ࡢᖹᆒ್࡛࠶ࡗࡓ 3.1 ms ᅛᐃࡋࠊ᥎ᐃᑐ㇟ࡢࣔࢹࣝࣃ࣓࣮ࣛࢱࡣ S ཬࡧ
Rmaxࡢ⪅ࡋࡓࠋ
㗹య⣔ᛂ⟅ࡢa Ἴࡢึᮇ㒊ศ㸦Cone a-wave㸧ࡣࠊ๓㏙ࡢᘧ (1) low-pass exponential filter
ࢆ⤌ࡳ㎸ࢇࡔࣔࢹࣝࢆ࣑࢚࢝ࣜࢫ࣓ࣥࢸࣥᘧ࡛⾲ࡋࡓᘧ (2)(26)ࢆᮞయ⣔ᛂ⟅ྠᵝࣇ ࢵࢸࣥࢢࡋࡓࠋ max 3 3
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(t > td) (2)R ࡣᖜ㸦μV㸧ࠊI ࡣග่⃭ᙉᗘ㸦log phot td-s㸧ࠊS ࡣឤᗘ㸦s-3(td-s)-1㸧ࠊR
maxࡣ᭱ᖜ㸦μV㸧ࠊ
t ࡣග่⃭ᚋ㛫㸦ms㸧ࠊtdࡣග่⃭ᚋࡢ▷㛫ࡢ㐜ࢀ㸦ms㸧ࢆࡑࢀࡒࢀ⾲ࡍࠋtdࡣᖖ࡞࢝
Rmaxࡢ⪅ࡋࡓࠋ
2.4. ║ᗏ᳨ᰝ
ERG ࡢྛホ౯࣏ࣥࢺ࡚ࠊERG グ㘓⤊┤ᚋ║ಽീ᳨║㙾㸦HEINE OMEGA 500;
HEINE Optotechnik GmbH & Co. KG㸧ࢆ⏝࠸࡚୧║ࡢ║ᗏࢆほᐹࡋࡓࠋ
2.5. ࢺ࢟ࢩࢥ࢟ࢿࢸࢡࢫ ᮏᐇ㦂࠾ࡅࡿERG ࡢኚ⾑୰ࢩࣝࢹࢼࣇࣝ⃰ᗘࡢ㛵ಀࢆㄪࡿࡇࠊཬࡧᮏᐇ㦂 ࡢ࢝ࢽࢡࢨ࡛ࣝࡢࢩࣝࢹࢼࣇࣝࡢ᭚㟢ࢆࣄࢺẚ㍑ࡍࡿࡇࢆ┠ⓗࡋ࡚ࠊࢺ࢟ࢩࢥ ࢟ࢿࢸࢡࢫ㸦toxicokinetics: TK㸧ゎᯒࢆ⾜ࡗࡓࠋࢩࣝࢹࢼࣇࣝࡢᢞᚋ ERG ࢆグ㘓ࡋ ࡓ㝿ࠊ⾑ᾮ⣙0.5 mL ࢆ䭇㟼⬦ࡽ᥇ྲྀࡋࡓࠋࡉࡽࠊࢩࣝࢹࢼࣇࣝᢞᚋ ERG ࢆグ 㘓ࡋ࡚ࡽ2 㐌㛫௨ୖ⤒㐣ࡋࡓᚋࠊࢩࣝࢹࢼࣇࣝ 1ࠊ3ࠊཬࡧ 10 mg/kg ⩌ࡢィ 12 ࢩ ࣝࢹࢼࣇࣝࢆᗘྠᵝᢞࡋࠊᢞ๓ࠊᢞ㛤ጞ 10 ཬࡧ 30 ศᚋࠊ2ࠊ4ࠊ7ࠊཬࡧ 24 㛫ᚋࠊྠᵝ᥇⾑ࡋࡓࠋ⾑ᾮࢆ11,200 g ཬࡧ 4Υ࡚ 5 ศ㛫㐲ᚰࡋ࡚ᚓࡽࢀࡓ⾑₢ࡣࠊ 㧗㏿ᾮయࢡ࣐ࣟࢺࢢࣛࣇ࣮ࡼࡿ⾑₢୰ࢩࣝࢹࢼࣇࣝ⃰ᗘ ᐃ⏝࠸ࡓࠋTK ࣃ࣓࣮ࣛ
ࢱࡋ࡚↓㝈㛫ࡲ࡛ࡢ⾑୰⃰ᗘ㸫㛫᭤⥺ୗ㠃✚㸦area under the concentration-time curve
up to infinity: AUC0-inf㸧ࢆ⟬ฟࡋࡓࠋ
ࡲࡓࠊᮏᐇ㦂ࡢ࢝ࢽࢡࢨ࡛ࣝࡢࢪࢦ࢟ࢩࣥࡢ᭚㟢ࢆࣄࢺẚ㍑ࡍࡿࡇࢆ┠ⓗࡋ࡚ࠊ TK ゎᯒࢆ⾜ࡗࡓࠋࢪࢦ࢟ࢩࣥᢞ 1 ࣧ᭶ᚋࡢ ERG ࢆグ㘓ࡋ࡚ࡽ 10 ᪥㛫௨ୖ⤒㐣ࡋࡓᚋࠊ
ࢪࢦ࢟ࢩࣥ⩌ࡢ 4 ࢪࢦ࢟ࢩࣥࢆᗘྠᵝᢞࡋࡓࠋᢞ๓ࠊᢞ㛤ጞ 20 ཬࡧ 40 ศ
ᚋࠊ1ࠊ2ࠊ4ࠊ7ࠊ24ࠊ48ࠊཬࡧ 72 㛫ᚋࠊ⾑ᾮ⣙ 0.6 mL ࢆ䭇㟼⬦ࡽ᥇ྲྀࡋࡓࠋ⾑ᾮ
28 2.6. ⤫ィᏛⓗゎᯒ ᢞ๓ᚋ࠾ࡅࡿࢩࣝࢹࢼࣇࣝ⩌ࡢྛࣔࢹࣝࣃ࣓࣮ࣛࢱࡣᖹᆒ್sᶆ‽೫ᕪ㸦SD㸧࡛ ⾲ࡋࠊࣃ࣓ࣛࢺࣜࢵࢡDunnett ᳨ᐃࢆ⏝࠸࡚፹యᑐ↷⩌ẚ㍑ࡋࡓࠋࢪࢦ࢟ࢩࣥᢞᚋࡢྛ ࣔࢹࣝࣃ࣓࣮ࣛࢱࡣᖹᆒ್sᶆ‽೫ᕪ㸦SD㸧࡛⾲ࡋࠊᑐᛂࡢ࠶ࡿ t ᳨ᐃࢆ⏝࠸࡚ᢞ๓್ ࠊᑐᛂࡢ࡞࠸t ᳨ᐃࢆ⏝࠸࡚፹యᑐ↷⩌ࡑࢀࡒࢀẚ㍑ࡋࡓࠋP<0.05 ࢆࡶࡗ࡚⤫ィᏛⓗ ᭷ព࡞ኚุ᩿ࡋࡓࠋ 3. ᐇ㦂⤖ᯝ 3.1. ࢩࣝࢹࢼࣇࣝ 3.1.1. ERG
Rod a-wave ཬࡧ Cone a-wave ࠾ࡅࡿἼᙧኚࡢ௦⾲ࢆࠊࡑࢀࡒࢀ Fig. 10 ཬࡧ Fig. 11
♧ࡍࠋࡲࡓࠊゎᯒࡋࡓྛࣔࢹࣝࣃ࣓࣮ࣛࢱࢆTab. 3 ♧ࡍࠋ
ᢞ┤ᚋࠊRod a-wave ࡛ࡣ 10 mg/kg ᢞ࡛ࠊCone a-wave ࡛ࡣ 3 ཬࡧ 10 mg/kg ᢞ࡛
ഴࡁࡢῶᑡࡀほᐹࡉࢀࡓࠋࡇࢀࡽࡢἼᙧኚࡣࠊᢞ24 㛫ᚋࡣᢞ๓ྠ⛬ᗘࡲ࡛ᅇ ࡋࡓࠋ ࣇࢵࢸࣥࢢࡢ⤖ᯝࠊ᥎ᐃἼᙧࡢᐇἼᙧࡽࡢ㢧ⴭ࡞㐓⬺ࡣ࠸ࡎࢀࡢື≀࡛ࡶㄆࡵࡽ ࢀࡎࠊ๓⠇㸦ᮦᩱཬࡧ᪉ἲ㸧࡚㏙ࡓ᮲௳࡚Ⰻዲ࡞ࣇࢵࢸࣥࢢࡀᚓࡽࢀࡓࡇࡀ ☜ㄆࡉࢀࡓࠋRod a-wave ࡛ࡣ S ࡢῶᑡࡀㄆࡵࡽࢀࠊᢞ┤ᚋ࠾ࡅࡿࢩࣝࢹࢼࣇࣝ 10 mg/kg ⩌ࡢ S ࡣ፹యᑐ↷⩌ẚ㍑ࡋ࡚᭷ព࡞ప್㸦P<0.01㸧ࢆ♧ࡋࡓࠋCone a-wave ࡛ࡣᢞ ⏝㔞㛵㐃ࡋࡓ S ࡢῶᑡࡀㄆࡵࡽࢀࡓࠋ፹యᑐ↷⩌ẚ㍑ࡋ࡚ࠊᢞ┤ᚋ࠾ࡅࡿࢩࣝ ࢹࢼࣇࣝ3 ཬࡧ 10 mg/kg ⩌ࡢ S ࡣ᭷ព࡞ప್㸦ࡶ P<0.01㸧࡛࠶ࡗࡓࠋᢞ 24 㛫ᚋ ࡛ࡣࠊ፹యᑐ↷⩌ࢩࣝࢹࢼࣇࣝ⩌ࡢ㛫᫂ࡽ࡞ S ࡢᕪࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ୍᪉ࠊ Rmaxࡣ᫂ࡽ࡞ኚࡣㄆࡵࡽࢀ࡞ࡗࡓࠋୖグࡢERG ࡢኚࠊ᫂ࡽ࡞㞤㞝ᕪࡣほᐹ ࡉࢀ࡞ࡗࡓࠋ
3.1.2. ║ᗏ᳨ᰝ ࠸ࡎࢀࡢほᐹ࣏ࣥࢺ࠾࠸࡚ࡶࠊ║ᗏ᫂ࡽ࡞␗ᖖࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ 3.1.3. ࢺ࢟ࢩࢥ࢟ࢿࢸࢡࢫ ࢩࣝࢹࢼࣇࣝᢞᚋࡢ⾑₢୰ࢩࣝࢹࢼࣇࣝ⃰ᗘཬࡧTK ࣃ࣓࣮ࣛࢱࢆ Tab. 4 ♧ࡍࠋ ࢩࣝࢹࢼࣇࣝᢞ┤ᚋࡢERG グ㘓㸦ᢞ㛤ጞᚋ 42-51 ศ㸧ࡢ⾑₢୰ࢩࣝࢹࢼࣇࣝ ⃰ᗘࡣࠊ1ࠊ3ࠊཬࡧ 10 mg/kg ⩌࡛ࡑࢀࡒࢀ 378 s 82.9ࠊ1190 ± 181ࠊཬࡧ 3610 ± 639 ng/mL ࡛࠶ࡗࡓࠋࢩࣝࢹࢼࣇࣝᢞ 24 㛫ᚋࡢ ERG グ㘓㸦ᢞ㛤ጞᚋ 24 㛫 34-56 ศ㸧ࡢ ⾑₢୰ࢩࣝࢹࢼࣇࣝ⃰ᗘࡣࠊ1 ཬࡧ 3 mg/kg ⩌࡛ࡣ᳨࡛ฟ㝈⏺㸦1 ng/mL㸧ࢆୗᅇࡗ࡚ ࠾ࡾࠊ10 mg/kg ⩌࡛ࡣ 1.54 ± 1.09 ng/mL ࡛࠶ࡗࡓࠋࡲࡓࠊࢩࣝࢹࢼࣇࣝࡢ༢ᅇᢞᚋࡢ AUC0-infࡣࠊ1ࠊ3ࠊཬࡧ 10 mg/kg⩌࡛ࡑࢀࡒࢀ 1000 ± 121ࠊ4250 ± 748ࠊཬࡧ 12900± 2020 ng∙h/mL ࡛࠶ࡗࡓࠋ 3.2. ࢪࢦ࢟ࢩࣥ 3.2.1. ERG
Rod a-wave ཬࡧ Cone a-wave ࠾ࡅࡿἼᙧኚࡢ௦⾲ࢆࠊࡑࢀࡒࢀ Fig. 12 ཬࡧ Fig. 13
♧ࡍࠋࡲࡓࠊゎᯒࡋࡓྛࣔࢹࣝࣃ࣓࣮ࣛࢱࢆTab. 5 ♧ࡍࠋ
Rod a-wave ࡣࠊᢞᚋ᫂ࡽ࡞ኚࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ୍᪉ࠊCone a-wave ࡣᢞ
24 㛫ᚋ㢧ⴭ࡞ῶᙅࢆ♧ࡋࡓࠋࣇࢵࢸࣥࢢࡢ⤖ᯝࠊ᥎ᐃἼᙧࡢᐇἼᙧࡽࡢ㢧ⴭ࡞ 㐓⬺ࡣ࠸ࡎࢀࡢື≀࡛ࡶㄆࡵࡽࢀࡎࠊ๓⠇㸦ᮦᩱཬࡧ᪉ἲ㸧࡚㏙ࡓ᮲௳࡚Ⰻዲ࡞ࣇ
ࢵࢸࣥࢢࡀᚓࡽࢀࡓࡇࡀ☜ㄆࡉࢀࡓࠋCone a-wave ࡛ࡣ Rmaxࡢῶᑡࡀㄆࡵࡽࢀࡓࠋࢪ
30 ࡣㄆࡵࡽࢀ࡞ࡗࡓࠋୖグࡢERG ࡢኚࠊ᫂ࡽ࡞㞤㞝ᕪࡣほᐹࡉࢀ࡞ࡗࡓࠋ 3.2.2. ║ᗏ᳨ᰝ ࠸ࡎࢀࡢほᐹ࣏ࣥࢺ࠾࠸࡚ࡶࠊ║ᗏ᫂ࡽ࡞␗ᖖࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ 3.2.3. ࢺ࢟ࢩࢥ࢟ࢿࢸࢡࢫ ࢪࢦ࢟ࢩࣥᢞᚋࡢ⾑Ύ୰ࢪࢦ࢟ࢩࣥ⃰ᗘཬࡧTK ࣃ࣓࣮ࣛࢱࢆ Tab. 6 ♧ࡍࠋ ࢪࢦ࢟ࢩࣥࡢ༢ᅇᢞᚋࡢAUC0-infࡣ334.4 ± 65.43 ng∙h/mL ࡛࠶ࡗࡓࠋ 4. ⪃ᐹ
Hood ࡽࡢ⪃ࡋࡓ a-wave analysis(27)࡛ࡣࠊᮞయ࠶ࡿ࠸ࡣ㗹య⏤᮶ࡍࡿERG ࡢ a Ἴึᮇ
㒊ศᩘᘧࣔࢹࣝࢆࡑࢀࡒࢀࣇࢵࢸࣥࢢࡍࡿࡇ࡛ࠊᮞయ࠶ࡿ࠸ࡣ㗹యࡢᶵ⬟㛵ࡍ ࡿࣔࢹࣝࣃ࣓࣮ࣛࢱ࡛࠶ࡿS㸦ឤᗘࣃ࣓࣮ࣛࢱ㸧ཬࡧ Rmax㸦᭱ᖜࣃ࣓࣮ࣛࢱ㸧ࡀࡑࢀࡒ ࢀồࡵࡽࢀࡿࠋS ࡣど⣽⬊እ⠇࠾ࡅࡿගሗኚࡢ㐣⛬ࢆᫎࡍࡿ⪃࠼ࡽࢀ࡚࠸ࡿࠋ୍ ᪉ࠊRmaxࡣྛど⣽⬊ࡢ᭱ᛂࡢ⥲ࢆពࡍࡿࡉࢀࡿࠋ Hood ࡽ(27)ࡼࡿࠊS ࡢῶᑡࡽࡣࠊࡋ࡚ど⣽⬊እ⠇࡛ࡢගሗኚࡢ␗ᖖࡀ♧၀ ࡉࢀࡿࠋ࠼ࡤࠊ⥙⭷୰ᚰ㟼⬦㛢ሰ➼ࡢど⣽⬊ࡀప㓟⣲࡞ࡿែࡢࣄࢺࡢᝈ⪅࡛ࡣࠊ Rmaxࡢኚࢆకࢃࡎ S ࡀ㑅ᢥⓗῶᑡࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ(31)ࠋࡇࢀࡣࠊప㓟⣲ ࡼࡗ࡚㉳ࡁࡿ௦ㅰᛶࢩࢻ࣮ࢩࢫࡼࡾࠊど⣽⬊እ⠇࡛ࡢගሗኚࡀḟⓗᙳ㡪ࢆཷ ࡅ࡚࠸ࡿࡶࡢ᥎ᐹࡉࢀ࡚࠸ࡿࠋ ୍᪉ࠊRmaxࡢῶᑡࡣࠊࡋ࡚ど⣽⬊ࡢᩘⓗῶᑡཪࡣど⣽⬊እ⠇ࡢ▷ᑠ➼ࡢど⣽⬊ࡢჾ ㉁ⓗ࡞⌮ኚࢆ♧၀ࡍࡿࡶࡢ⪃࠼ࡽࢀ࡚࠸ࡿ(27)ࠋ࠼ࡤࠊど⣽⬊ࡢኚᛶᛶᝈ࡛࠶ࡿ
⥙⭷Ⰽ⣲ኚᛶࡢᝈ⪅࡛ࡣࠊS ẚ㍑ࡍࡿ Rmaxࡀ㢧ⴭῶᑡࡍࡿ(68)ࠋࡲࡓࠊa-wave analysis
㑅ᢥⓗῶᑡࡍࡿࡇࡀ▱ࡽࢀࡿ(27)ࠋࡇࡢኚࡢཎᅉࡀど⣽⬊ࡢჾ㉁ⓗኚ࡛ࡣ࡞࠸ࡇ ࡣ᫂ࡽ࡛࠶ࡿࠋࡇࢀࡣࠊ⫼ᬒගࡼࡗ࡚ど⣽⬊እ⠇ࡢ⎔≧ࢢࣀࢩ୍ࣥࣜࣥ㓟㸦cyclic guanosine monophosphate: cGMP㸧౫Ꮡᛶ Na+ࢳࣕࢿࣝࡢ㛤ཱྀࡀῶᑡࡋࠊࡑࢀࡼࡾྛど⣽⬊ ࠾࠸࡚ᬯ⣽⬊እᾮࢆෆ⠇ࡽእ⠇ྥࡗ࡚ὶࢀࡿ㟁ὶ࡛࠶ࡿᬯ㟁ὶࡀ୍ⓗῶ ᑡࡋࡓ⤖ᯝ㸦⭷㟁ࡢ㐣ศᴟࡼࡿගᛂ⟅ࡢᅽ⦰㸧࡛࠶ࡿ᥎ᐹࡉࢀ࡚࠸ࡿ(25, 27)ࠋ ᮏᐇ㦂ࡢࢧࣝ࠾ࡅࡿࢩࣝࢹࢼࣇࣝ༢ᅇᢞᚋࡢAUC0-infࡣ1ࠊ3ࠊཬࡧ 10 mg/kg ⩌࡛ ࡑࢀࡒࢀ1000 ± 121ࠊ4250 ± 748ࠊཬࡧ 12900 ± 2020 ng∙h/mL ࡛࠶ࡗࡓࠋ୍᪉ࠊᖖே⮫ ᗋ⏝㔞㸦50 mg㸧ࡢࢩࣝࢹࢼࣇࣝࢆ༢ᅇ⤒ཱྀᢞࡋࡓ㝿ࡢ⾑₢୰ࢩࣝࢹࢼࣇࣝࡢ AUC0-inf ࡀ700 ng∙h/mL ࡛࠶ࡗࡓࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ(76)ࠋࡑࡢࡓࡵࠊ⮫ᗋ⏝㔞㸦50 mg/men㸧ࡢᢞ ࢆཷࡅࡓࣄࢺẚ㍑ࡋ࡚ࠊᮏᐇ㦂ࡢࢧ࡛ࣝࡢࢩࣝࢹࢼࣇࣝࡢ㌟᭚㟢ࡣࠊ1 mg/kg ⩌࡛ ࡣྠ⛬ᗘࠊ3 ཬࡧ 10 mg/kg ⩌࡛ࡣࡑࢀࡒࢀ⣙ 6 ཬࡧ 18 ಸ࡛࠶ࡾࡁࡃୖᅇࡗ࡚࠸ࡓ⪃࠼ ࡓࠋ ຨ㉳ཬࡧ⫵ື⬦ᛶ⫵㧗⾑ᅽࡢ⒪⸆ࡋ࡚⏝࠸ࡽࢀ࡚࠸ࡿࢩࣝࢹࢼࣇࣝࡣࠊᝈ ⪅୍㐣ᛶࡢどぬ㞀ᐖࢆㄏⓎࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ(33)ࠋࡲࡓࢩࣝࢹࢼࣇࣝࡣࠊど⣽⬊ እ⠇࠾ࡅࡿගሗኚࡢ㔜せ࡞㓝⣲ࡢ୍ࡘ࡛࠶ࡿ PDE6 ᑐࡍࡿ㜼ᐖస⏝ࢆ᭷ࡍࡿ(77)ࠋ ࡑࡢࡓࡵࠊୖグࡢどぬ㞀ᐖࡣࢩࣝࢹࢼࣇࣝࡼࡿど⣽⬊ࡢPDE6 㜼ᐖ㛵㐃ࡋࡓࡶࡢ୍ ⯡ⓗ⪃࠼ࡽࢀ࡚࠸ࡿ(33)ࠋࡇࢀࡽࡢࡇࡽࠊa-wave analysis ࡼࡗ᳨࡚ฟࡉࢀࡿࢩࣝࢹࢼ ࣇࣝࡼࡿど⣽⬊ᶵ⬟ࡢኚࡋ࡚ࡣࠊගሗኚࡢ㐣⛬ࢆᫎࡍࡿឤᗘࣃ࣓࣮ࣛࢱ㸦S㸧 ࡢ㑅ᢥⓗ࡞ῶᑡࡀணࡉࢀࡿࠋ ᮏᐇ㦂ࡢ⤖ᯝࠊࢩࣝࢹࢼࣇࣝᢞ┤ᚋS ࡢῶᑡࡀㄆࡵࡽࢀࠊCone a-wave ࡛ࡣ 3 mg/kg ௨ୖࡢᢞ⩌࡛ࠊRod a-wave ࡛ࡣ 10 mg/kg ᢞ⩌࡛ῶᑡࡋࡓࠋ୍᪉ࠊ᭱ᖜࣃ࣓࣮ࣛࢱ
32
↷⩌ྠ⛬ᗘ࡛࠶ࡗࡓࡇࡽࠊS ࡢῶᑡࡀࢩࣝࢹࢼࣇࣝࡢ⾑₢୰⃰ᗘ㛵㐃ࡋࡘྍ㏫
ⓗ࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋࡉࡽࠊᮏᐇ㦂࡚S ࡀ Rod a-wave ࡼࡾࡶ Cone a-wave ࡛ࡼࡾ
ప⏝㔞ࡽኚࡋࡓࡇࡽࠊࢩࣝࢹࢼࣇࣝࡀᮞయẚ㍑ࡋ࡚㗹యࡢᶵ⬟ࢆࡼࡾᙉࡃ㞀 ᐖࡍࡿࡇࡀ♧၀ࡉࢀࡓࠋࡇࡢࡇࡣࠊ⢭〇ࡋࡓど⣽⬊⏤᮶ࡢPDE6 ᑐࡍࡿࢩࣝࢹࢼࣇ ࣝࡢ㜼ᐖάᛶࢆㄪࡓࡇࢁ㗹యࡀᮞయࡼࡾࡶࡼࡾప⃰ᗘࡽ㜼ᐖࢆཷࡅࡓࡍࡿ in vitro ᐇ㦂ࡢሗ࿌(77)ࡽࡶᨭᣢࡉࢀࡿࠋ௨ୖࡼࡾࠊᮏᐇ㦂᳨࡚ฟࡉࢀࡓឤᗘࣃ࣓࣮ࣛࢱ S ࡢ㑅 ᢥⓗࡘྍ㏫ⓗ࡞ῶᑡࡣࠊࢩࣝࢹࢼࣇࣝࡼࡿど⣽⬊ࡢPDE6 㜼ᐖస⏝ࢆᫎࡋࡓኚ ⪃࠼ࡓࠋ ᮏᐇ㦂ࡢࢧࣝ࠾ࡅࡿࢪࢦ࢟ࢩࣥ༢ᅇᢞᚋࡢAUC0-infࡣ334.4 ± 65.43 ng∙h/mL ࡛࠶ࡗࡓࠋ ୍᪉ࠊᖖே⮫ᗋ⏝㔞㸦0.25 mg/men/day㸧ࡢࢪࢦ࢟ࢩࣥࢆ⤒ཱྀᢞࡋࡓࡇࢁࠊᐃ ᖖ≧ែ࠾ࡅࡿᢞ㛫㝸㸦24 㛫㸧ẖࡢ⾑₢୰⃰ᗘ㸫㛫᭤⥺ୗ㠃✚㸦AUC during dosing interval at steady state: AUCtau, ss㸧ࡢᖹᆒ್ࡀ15.4 ng͌h/mL ࡛࠶ࡗࡓࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ(17)ࠋ
ࡑࡢࡓࡵࠊᮏᐇ㦂࡛ࡢࢪࢦ࢟ࢩࣥࡢ AUC ࡣࠊ⮫ᗋ⏝㔞ࡢᢞࢆཷࡅࡓࣄࢺࡢ⣙ 22 ಸ࡛࠶ ࡗࡓ⪃࠼ࡓࠋ ࣄࢺ࡛どぬ㞀ᐖࢆㄏⓎࡍࡿࢪࢠࢱࣜࢫࡀࠊど⣽⬊ෆ⠇ࡢNa+-K+ATPase ࡢάᛶࢆ㜼ᐖࡍࡿ ࡇࡣྂࡃࡽ▱ࡽࢀ࡚࠸ࡿ(4)ࠋࡲࡓࠊど⣽⬊ࡢNa+-K+ATPase ࡣࠊ⣽⬊ෆ K+ࢆ䬦ࡳධࢀ ࡿࡇ࡛ᬯ㟁ὶࡢⓎ⏕㔜せ࡞ᙺࢆᯝࡓࡍࠋMadreperla ࡽࡣࠊ༢㞳ࡋࡓࢺࣛࣇࢧࣥࢩࣙ࢘ ࢘࢜ࡢど⣽⬊࠾ࡅࡿගᛂ⟅ࡀࢪࢦ࢟ࢩࣥࡢ⃰ᗘ౫Ꮡⓗῶᙅࡋࠊࡑࡢῶᙅࡣᮞయẚ㍑ ࡋ࡚㗹య࡛㢧ⴭ࡛࠶ࡗࡓࡇࢆሗ࿌ࡋࡓ(39)ࠋࡇࡢ⌧㇟ࢆ Madreperla ࡽࡣࠊࢪࢦ࢟ࢩࣥࡀど ⣽⬊ࠊ≉㗹యࡢNa+-K+ATPase άᛶࢆᙉࡃ㜼ᐖࡍࡿࡇ࡛ᬯ㟁ὶࡀῶᑡࡋࠊࡑࡢ⤖ᯝࡋ ࡚㗹యඃ࡞ど⣽⬊ࡢᶵ⬟㞀ᐖࡀㄏⓎࡉࢀ࡚࠸ࡿ⪃࠼ࡓࠋࡇࡢゎ㔘ᚑ࠺ࠊࢪࢦ࢟ࢩ ࣥࡼࡿど⣽⬊ෆ⠇ࡢNa+-K+ATPase 㜼ᐖࡣࠊᬯ㟁ὶࢆῶᑡࡉࡏࡿࡀእ⠇࡛ࡢගሗኚࡢ 㐣⛬ࡣ≉ᙳ㡪ࢆཬࡰࡉ࡞࠸࠸࠺Ⅼ࡛ࠊ๓㏙ࡢ⫼ᬒගࡼࡿcGMP ౫Ꮡᛶ Na+ࢳࣕࢿࣝ ࡢ㛤ཱྀῶᑡ㢮ఝࡋ࡚࠸ࡿࠋࡲࡓࠊ๓㏙ࡢࡼ࠺in vitro ࡚☜ㄆࡉࢀ࡚࠸ࡿࢪࢦ࢟ࢩࣥ ࡼࡿど⣽⬊ࡢᢚไࡣ㗹య㢧ⴭ࡞ࡶࡢ࡛࠶ࡿࠋࡇࢀࡽࡢࡇࡽࠊa-wave analysis ࡼࡗ࡚
᳨ฟࡉࢀࡿࢪࢦ࢟ࢩࣥࡼࡿど⣽⬊ᶵ⬟ࡢኚࡋ࡚ࡣࠊCone a-wave ࠾ࡅࡿ᭱ᖜࣃ
࣓࣮ࣛࢱ㸦Rmax㸧ࡢ㢧ⴭ࡞ῶᑡࡀணࡉࢀࡿࠋ
ᮏᐇ㦂ࡢ⤖ᯝࡣୖグࡢ௬ㄝ୍⮴ࡍࡿࡶࡢ࡛࠶ࡾࠊRod a-wave ࠾ࡅࡿ S ཬࡧ RmaxࠊCone
a-wave ࠾ࡅࡿ S ཬࡧ Rmaxࡢ4 ࡘࡢࣔࢹࣝࣃ࣓࣮ࣛࢱࡢ࡞࡛ࠊࢪࢦ࢟ࢩࣥࡢᢞᚋ Cone
a-wave ࠾ࡅࡿ Rmaxࡢࡳࡀ᫂ࡽ࡞ῶᑡࢆ♧ࡋࡓࠋࡇࡢኚࡣᅇࡋࡓࡓࡵࠊࡑࡢཎᅉࡀ ど⣽⬊ࡢჾ㉁ⓗኚ࠶ࡿࡣ⪃࠼㞴࠸ࠋ௨ୖࡼࡾࠊᮏᐇ㦂᳨࡚ฟࡉࢀࡓCone a-wave ࠾ࡅࡿྍ㏫ⓗ࡞Rmaxࡢῶᑡࡣࠊࢪࢦ࢟ࢩࣥࡼࡿど⣽⬊ࡢNa+-K+ATPase 㜼ᐖస⏝ࢆᫎࡋ ࡓኚ⪃࠼ࡓࠋ 5. ᑠᣓ ࢩࣝࢹࢼࣇࣝཬࡧࢪࢦ࢟ࢩࣥࢆࡑࢀࡒࢀᢞࡋࡓ࢝ࢽࢡࢨ࡛ࣝ a-wave analysis ࢆᐇ ࡋࡓࠋࡑࡢ⤖ᯝࠊࡑࢀࡒࢀឤᗘࣃ࣓࣮ࣛࢱ S ཬࡧ᭱ᖜࣃ࣓࣮ࣛࢱ Rmaxࡢ㑅ᢥⓗ࡞ῶ ᑡࡀ᳨ฟࡉࢀࡓࠋࡇࡢ S ཬࡧ Rmaxࡢኚࡣࠊࡑࢀࡒࢀࢩࣝࢹࢼࣇࣝࡼࡿど⣽⬊እ⠇ࡢ PDE6 㜼ᐖస⏝ཬࡧࢪࢦ࢟ࢩࣥࡼࡿど⣽⬊ෆ⠇ࡢ Na+-K+ATPase 㜼ᐖస⏝ࡽணࡉࢀࡿ ࣔࢹࣝࣃ࣓࣮ࣛࢱࡢኚ୍⮴ࡍࡿࡶࡢ࡛࠶ࡗࡓࠋ௨ୖࡼࡾࠊ࢝ࢽࢡࢨ࡛ࣝࡢ a-wave analysis ࡼࡗ࡚ࠊࢩࣝࢹࢼࣇࣝཬࡧࢪࢦ࢟ࢩࣥࡼࡿ୍ḟࢽ࣮ࣗࣟࣥ㸦ど⣽⬊㸧ࡢᶵ⬟ ᑐࡍࡿ᪤▱ࡢస⏝ࢆᫎࡍࡿኚࢆ᳨ฟࡍࡿࡇࡀྍ⬟࡛࠶ࡿࡇࡀ♧ࡉࢀࡓࠋ
34
FIGURE 7. A schematic representation of the phototransduction cascade at the outer segment of
the rod photoreceptors. (A) In the dark, cGMP-gated cation channel is open. (B) Light transduces the visual pigment via the following enzyme cascade: photons - rhodopsin - transducin - phosphodiesterase type 6 - closes a membrane bound cGMP-gated cation channel. Abbreviations: R, rhodopsin; T, transducin; PDE6, phosphodiesterase type 6.
FIGURE 8. Recovery of the response amplitude for the second stimulation in a pair of bright
flashes in cynomolgus monkeys. Pairs of identical white bright flashes of 4.0 log scot td-s were delivered at a range of inter-flash intervals. Symbols plot the response amplitude of the a-wave for the second flash, as a fraction of the response amplitude of the a-wave for single flashes under fully dark-adapted conditions at a measurement time of 8.0 ms. Each symbol represents the mean s SD of three animals. The lower panel shows the same results as the upper panel, but on a faster time scale and an enlarged vertical scale. One component of the response recovered rapidly, reaching a plateau of approximately 30% of the maximal ‘rod + cone’ level about 0.3 s, and it was interpreted
36
FIGURE 9. The schema of obtaining the rod-isolated response in cynomolgus monkeys. (A)
Combined rod-cone responses (thin black line) were elicited by white bright flashes in fully dark-adapted state. Thereafter, cone responses (thick gray line) were elicited by flashes of the same intensity presented 0.5 s after a rod-saturating conditioning flash at 4.0 log scot td-s. The difference between these two responses is the rod-isolated response (thick black line). The traces represent the responses elicited by light flashes at 4.3 log scot td-s from one representative animal. (B) The first 20 ms of the records in panel A is shown. The traces of the combined rod-cone response and the cone response represent averages of three and six responses, respectively.
FIGURE 10. Typical waveforms of the rod a-wave in vehicle- and sildenafil-treated cynomolgus
monkeys. The solid curves in each panel represent the rod a-waves, which were derived by subtracting cone responses from combined rod-cone responses as described in the text. The dotted lines are the curves fit from the equation (1). The responses obtained before dosing (gray trace) are superimposed on those obtained after dosing (black trace) in the middle (After , immediately after) and right (Recovery, 24 hours after) panels.
38
FIGURE 11. Typical waveforms of the cone a-wave in vehicle- and sildenafil-treated cynomolgus
monkeys. The solid curves in each panel represent the cone a-waves, which were derived by means of the paired-flash technique as described in the text. Each trace represents the average of six responses. The dotted lines are the curves fit from the equation (2). The responses obtained before dosing (gray trace) are superimposed on those obtained after dosing (black trace) in the middle (After, immediately after) and right (Recovery, 24 hours after) panels.
FIGURE 12. Typical waveforms of the rod a-wave in vehicle- and digoxin-treated cynomolgus
monkeys. The solid curves in each panel represent the rod a-waves, which were derived by subtracting cone responses from combined rod-cone responses as described in the text. The dotted lines are the curves fit from the equation (1). The responses obtained before dosing (gray trace) are
40
FIGURE 13. Typical waveforms of the cone a-wave in vehicle- and digoxin-treated cynomolgus
monkeys. The solid curves in each panel represent the cone a-waves, which were derived by means of the paired-flash technique as described in the text. Each trace represents the average of six responses. The dotted lines are the curves fit from the equation (2). The responses obtained before dosing (gray trace) are superimposed on those obtained after dosing (black trace) in the middle (After, 24 hours after) and right (Recovery, one month after) panels.
TABLE 3. Effects of Sildenafil on the A-wave Analysis Parameters in Cynomolgus Monkeys
Data are expressed as the mean ± SD of four animals. 1) Recorded immediately after dosing.
2) Recorded 24 hours after dosing.
* P<0.05, ** P<0.01; significantly different in comparison with the vehicle-treated group by the Dunnett's test. Rod a-wave Log S (s-2(td-s)-1) 0 1.15 㼼 0.086 1.12 㼼 0.108 1.20 㼼 0.038 1 1.17 㼼 0.141 1.14 㼼 0.152 1.12 㼼 0.113 3 1.22 㼼 0.078 1.15 㼼 0.073 1.21 㼼 0.074 10 1.12 㼼 0.057 0.77 㼼 0.167 1.13 㼼 0.116 Log Rmax (μV) 0 2.07 㼼 0.222 2.10 㼼 0.189 2.11 㼼 0.190 1 2.06 㼼 0.040 2.12 㼼 0.068 2.11 㼼 0.081 3 2.11 㼼 0.065 2.14 㼼 0.059 2.13 㼼 0.084 10 2.04 㼼 0.047 2.08 㼼 0.034 2.06 㼼 0.093 Cone a-wave Log S (s-3(td-s)-1) 0 3.81 㼼 0.120 3.77 㼼 0.113 3.77 㼼 0.137 1 3.79 㼼 0.101 3.56 㼼 0.091 3.70 㼼 0.107 3 3.80 㼼 0.044 3.12 㼼 0.166 3.83 㼼 0.062 10 3.84 㼼 0.084 2.56 㼼 0.083 3.92 㼼 0.085 Log Rmax (μV) 0 1.72 㼼 0.181 1.75 㼼 0.140 1.76 㼼 0.144 1 1.74 㼼 0.050 1.80 㼼 0.044 1.79 㼼 0.046 3 1.75 㼼 0.048 1.84 㼼 0.102 1.77 㼼 0.033 10 1.68 㼼 0.101 1.71 㼼 0.121 1.68 㼼 0.130 Recovery 2) Dose (mg/kg) Before After 1) ** ** **
42
TABLE 4. Plasma Sildenafil Concentrations and Toxicokinetic Parameters After Sildenafil Dosing
Data are expressed as the mean ± SD of four animals. NC, Not calculated.
* Individual values were below the lower limit of quantitation (1 ng/mL).
Pre 10 min 30 min 2 h 4 h 7 h 24 h
1070 515 57.5 10.7 1.16 1000 1070 0.835 ±66.8 ±90.4 ±18.7 ±5.03 ±0.836 ±114 ±66.8 ±0.211 5610 1630 188 21.2 2.46 4250 5610 0.785 ±1670 ±558 ±59.3 ±3.07 ±0.557 ±748 ±1670 ±0.0860 15600 5880 634 86.6 11.3 12900 15600 2.65 ±2610 ±955 ±276 ±81.0 ±5.83 NC* ±2020 ±2610 ±2.09 10 NC* NC* NC* t1/2 (h) Time After the Start of 10-min Infusion
1 3 NC* NC* Dose (mg/kg) Plasma Concentration (ng/mL) AUC0-inf (ng䌭h/mL) Cmax (ng/mL)
TABLE 5. Effects of Digoxin on the A-wave Analysis Parameters in Cynomolgus Monkeys
Data are expressed as the mean ± SD of four animals. 1) Recorded 24 hours after dosing.
2) Recorded one month after dosing.
* P < 0.05, ** P < 0.01; significantly different in comparison with the baseline value by the paired t-test.
͊ P < 0.05, ‡ P < 0.01; significantly different in comparison with the vehicle-treated group by the Student’s t-test. Rod a-wave Log S (s-2 (td-s)-1 ) 1.14 㼼 0.195 1.11 㼼 0.138 1.20 㼼 0.113 1.18 㼼 0.095 1.08 㼼 0.176 1.32 㼼 0.131 Log Rmax (μV) 2.00 㼼 0.153 2.00 㼼 0.161 2.03 㼼 0.097 1.97 㼼 0.137 2.00 㼼 0.158 2.03 㼼 0.111 Cone a-wave Log S (s-3 (td-s)-1 ) 3.74 㼼 0.114 3.75 㼼 0.108 3.85 㼼 0.098 3.88 㼼 0.120 3.86 㼼 0.228 3.90 㼼 0.222 Log Rmax (μV) 1.63 㼼 0.099 1.62 㼼 0.122 1.66 㼼 0.099 1.63 㼼 0.124 1.39 㼼 0.130 1.69 㼼 0.124 Digoxin-Treated Group Vehicle-Treated Group Recovery 2) Before After 1) Recovery 2) Before After 1)
44
TABLE 6. Serum Digoxin Concentrations and Toxicokinetic Parameters After Digoxin Dosing
Data are expressed as the mean ± SD of four animals. NC, Not calculated.
* Individual values were below the lower limit of quantitation (0.3 ng/mL).
Pre 20 min 40 min 1 h 2 h 4 h 7 h 24 h 48 h 72 h
41.5 52.1 22.5 9.7 5.5 4.8 2.7 2.0 1.4 334.4 52.7 44.8 ± 11.44 ± 24.97 ± 4.47 ± 2.19 ± 0.94 ± 0.62 ± 0.38 ± 0.45 ± 0.24 ± 65.43 ± 24.61 ± 25.00
t1/2
(h) Time After the Start of 40-min Infusion
0.1 NC* Dose (mg/kg) Serum Concentration (ng/mL) AUC0-inf (ng䌭h/mL) Cmax (ng/mL)
➨
3 ❶ ON-OFF response㸸ḟࢽ࣮ࣗࣟࣥ㸦ᴟ⣽⬊㸧ࡢホ౯
1. ᗎㄽ ⥙⭷ࡢḟࢽ࣮࡛ࣗࣟࣥ࠶ࡿᴟ⣽⬊ࡣࠊ⚄⤒ఏ㐩≀㉁࡛࠶ࡿࢢࣝࢱ࣑ࣥ㓟ࢆࡋ࡚ࠊ ど⣽⬊ࡽࡢどぬሗࢆཷࡅྲྀࡿࠋࡲࡓࠊ့ங㢮ࡢ⥙⭷࡛ࡣࠊど⣽⬊ࡣගࡢቑᑐࡋ࡚ 㐣ศᴟࢆ♧ࡍࠋ୍᪉ࠊᴟ⣽⬊ࡣගࡢቑᑐࡋ࡚ࡣ⬺ศᴟࢆࠊῶᑡᑐࡋ࡚ࡣ㐣ศᴟ ࢆ♧ࡍ ON ᆺᴟ⣽⬊ࠊࡑࡢ㏫ࡢᛂࢆ♧ࡍ OFF ᆺᴟ⣽⬊ࡢ 2 ✀㢮ࡀᏑᅾࡍࡿࡇࡀ ▱ࡽࢀࡿ(50)ࠋࡇࡢON ᆺཬࡧ OFF ᆺᴟ⣽⬊ࡢᛂࡢ㐪࠸ࡣࠊど⣽⬊ࡢࢩࢼࣉࢫ࠾ࡅ ࡿᶞ≧✺㉳Ꮡᅾࡍࡿࢢࣝࢱ࣑ࣥ㓟ཷᐜయࡢࢱࣉࡀ␗࡞ࡿࡇࡼࡿࡶࡢ࡛࠶ࡿࡀࠊࡇ ࢀࡣどぬሗఏ㐩࠾ࡅࡿ ON ⤒㊰ OFF ⤒㊰ࡀᴟ⣽⬊ࡢ࡛ࣞ࣋ࣝጞࡲࡿࡇࢆពࡍ ࡿ(50)ࠋ ගᑐࡍࡿᴟ⣽⬊ࡢ⭷㟁ࡢኚࡣࠊERG ࡛ࡣ a Ἴࡢᚋㄆࡵࡽࢀࡿ㝧ᛶἼ࡛࠶ࡿ b Ἴ㸦Fig. 4㸧᭱ࡶᫎࡉࢀࡿࠋࡑࡢࡓࡵࠊ୍⯡ⓗ ERG ࡢ b Ἴࡣᴟ⣽⬊ࡢᶵ⬟ᣦᶆࡋ ࡚⏝࠸ࡽࢀࡿࠋ୍᪉ࠊERG ࡣ㛝ග่⃭࡚ㄏⓎࡉࢀࡿࣇࣛࢵࢩࣗ ERG ࡀ୍⯡ⓗ࡛࠶ࡿࡀࠊᩘⓒ࣑ࣜ⛊㛫ᣢ⥆ࡍࡿ㛗㛫่⃭ࡼࡗ࡚ࡶERG ࡣㄏⓎࡉࢀࠊࡇࡢᛂ⟅ࡣ ON-OFF response
ࡤࢀࡿ(44)ࠋON-OFF response ࡛ࡣࠊගࡢቑᑐࡍࡿ⥙⭷ࡢᛂࡘࡲࡾ ON ᛂ⟅ࠊග
ࡢῶᑡᑐࡍࡿ⥙⭷ࡢᛂࡘࡲࡾOFF ᛂ⟅ࡀศ㞳ࡋ࡚グ㘓ࡉࢀࡿࡓࡵࠊࣇࣛࢵࢩࣗ ERG ࡛
ࡣᅔ㞴࡞ON ⤒㊰ OFF ⤒㊰ࢆศ㞳ࡋࡓホ౯ࡀྍ⬟࡞ࡿࠋᐇ㝿ࠊSieving ࡽࡣ≉␗ⓗ࡞
ࢩࢼࣉࢫ㐽᩿⸆ࢆ◪Ꮚయෆᢞࡋࡓ࣐࢝ࢡᒓ࡚ON-OFF response ࢆグ㘓ࡋࡓ⤖ᯝࡽࠊ㟋
