モノアミン受容体阻害による膵β細胞からのインスリン分泌抑制機序に関する研究
概要
༤ ኈ ㄽ ᩥ
ࣔࣀ࣑ࣥཷᐜయ㜼ᐖࡼࡿ⮅ β ⣽⬊ࡽࡢ
ࣥࢫࣜࣥศἪᢚไᶵᗎ㛵ࡍࡿ◊✲
ᗈᓥᏛᏛ㝔་ṑ⸆ಖᏛ◊✲⛉ ་ṑ⸆Ꮫᑓᨷ
㝔⸆Ꮫ◊✲ᐊ
ᖹᡂ ᖺᗘධᏛ 㛗⏣ 㯞ኸ
ᣦᑟᩍဨ ᯇᑿ ⿱ᙲ
┠ḟ
1
ᗎㄽ
ᮏㄽ
1. ࣁ࣒ࢫࢱ࣮⏤᮶⮅ β ⣽⬊ᰴ (HIT-T15) ⣽⬊࠾ࡅࡿྛࣔࣀ࣑ࣥཷᐜయ
ࡢⓎ⌧ゎᯒ
7
2. HIT-T15 ⣽⬊ࢆ⏝࠸ࡓࣥࢫࣜࣥศἪኚືࡢホ౯
10
3. HIT-T15 ⣽⬊ࡽࡢࣥࢫࣜࣥศἪ࠾ࡼࡰࡍ࢜ࣛࣥࢨࣆࣥࡢᙳ㡪
11
4. HIT-T15 ⣽⬊ࡽࡢࣥࢫࣜࣥศἪ࠾ࡼࡰࡍ࢜ࣛࣥࢨࣆࣥ 30 ᪥㛫᭚㟢
ࡢᙳ㡪
13
5. HIT-T15 ⣽⬊࠾ࡅࡿ࢜ࣛࣥࢨࣆࣥࡼࡿ⣽⬊ẘᛶࡢ☜ㄆ
15
6. HIT-T15 ⣽⬊ࡽࡢࣥࢫࣜࣥศἪ࠾ࡅࡿࢻࣃ࣑ࣥཷᐜయࡢ㛵
16
7. HIT-T15 ⣽⬊ࡽࡢࣥࢫࣜࣥศἪ࠾ࡅࡿࢭࣟࢺࢽࣥཷᐜయࡢ㛵
22
8. HIT-T15 ⣽⬊ࡽࡢࣥࢫࣜࣥศἪ࠾ࡅࡿࣄࢫࢱ࣑ࣥཷᐜయࡢ㛵
27
9. HIT-T15 ⣽⬊࠾ࡅࡿྛࣔࣀ࣑ࣥཷᐜయ㛵㐃⸆ࡼࡿ⣽⬊ẘᛶࡢ☜ㄆ
31
⤖ㄽ
33
ㅰ㎡
34
ᐇ㦂᪉ἲ
35
ㄽᩥ┠㘓
42
ཧ⪃ᩥ⊩
43
␎ྕ୍ぴ
AC
ADP
ANOVA
ATP
cAMP
cDNA
BSA
DAG
DNA
DMSO
DPA
DTP
EC50
EDTA
ELISA
Epac2
ER
FBS
GDP
GIP
GLP-1
GLU
GLUT
G6P
Ham’s F12K
HBSS
HRP
IP3
Kir
LMW
MARTA
mRNA
NMDA
PBS
PC
PCR
2-PEA
PIP2
PKA
PKC
PLC
PMS
PNGase
PVDF
SD
SDA
: adenylate cyclase
: adenosine diphosphate
: analysis of variance
: adenosine triphosphate
: cyclic adenosine monophosphate
: complementary deoxyribonucleic acid
: bovine serum albumin
: diacylglycerol
: deoxyribonucleic acid
: dimethyl sulfoxide
: dopamine partial agonist
: guanosine triphosphate
: half maximal effective concentration
: ethylenediaminetetraacetic acid
: enzyme-linked immuno sorbent assay
: exchange protein directly activated by cAMP 2
: endoplasmic reticulum
: fetal bovine serum
: guanosine diphosphate
: gastric inhibitory polypeptide
: glucagon-like peptide-1
: glucose
: glucose transporter
: glucose-6-phosphate
: Nutrient Mixture F-12 Ham Kaighn’s modification
: Hank’s balanced salts
: horseradish peroxidase
: inositol trisphosphate
: inward-rectifier potassium channel
: low molecular weight
: multi-acting receptor targeted antipsychotics
: messenger ribonucleic acid
: N-methyl-D-aspartate
: Dulbecco’s phosphate buffered saline
: prohormone convertase
: polymerase chain reaction
: 2-pyridylethylamine
: phosphatidylinositol bisphosphate
: protein kinase A
: protein kinase C
: phospholipase C
: N-methyl dibenzopyrazine methylsulfate
: peptide-N-glycosidase
: polyvinylidene difluoride
: standard deviation
: serotonin-dopamine antagonist
SDS-PAGE
SERCA
SNAP25
SUR
TAE
TBS
UV
VDCC
XTT
: polyacrylamide gel electrophoresis
: sarco/endoplasmic reticulum Ca2+-ATPase
: synaptosomal nerve-associated protein 25
: sulfonylurea receptor
: Tris-acetate-EDTA
: Tris buffered saline
: ultraviolet
: voltage-dependent Ca2+ channel
: sodium 3’-[1(-phenylaminocarbonyl)-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzene
sulfonic acid hydrate
ᗎㄽ
⤫ྜኻㄪࡣ 10㹼30 ௦ࡢ㟷ᖺᮇዲⓎࡍࡿෆᅉᛶ⢭⚄ᝈࡢ୍✀࡛࠶ࡿࠋMcGrath ࡽࡢㄪᰝ
ࡼࡿࠊ⤫ྜኻㄪࡢ⏕ᾭ⨯ᝈ⋡ࡣ 0.7%ሗ࿌ࡉࢀ࡚࠸ࡿ (1)ࠋࡲࡓࠊཌ⏕ປാ┬ࡼࡿ 2017
ᖺࡢᝈ⪅ㄪᰝ࡛ࡣࠊ⤫ྜኻㄪࡼࡾ་⒪ᶵ㛵ࢆཷデࡋ࡚࠸ࡿᝈ⪅ࡢᩘࡣ 79.2 ே┦ᙜࡍࡿ
᥎ ࡉࢀ࡚࠸ࡿ (2)ࠋ⤫ྜኻㄪࡢែࡣ᫂ࡽࡉࢀ࡚࠸࡞࠸ࡇࢁࡶከ࠸ࡀࠊࡇࢀࡲ࡛ࠊ⤫
ྜኻㄪࡢ⒪⏝࠸ࡽࢀࡿᢠ⢭⚄⸆ࡢ⮫ᗋ⏝㔞ࢻࣃ࣑ࣥ D2 ཷᐜయࡢ㐽᩿స⏝ᑐࡍࡿຊ౯
ࡢ┦㛵ᛶࡀ㧗࠸ࡇࡸ (3)ࠊࢻࣃ࣑ࣥཷᐜయ่⃭⸆࡛࠶ࡿࣥࣇ࢙ࢱ࣑ࣥࡸ࣓ࢱࣥࣇ࢙ࢱ࣑ࣥࡢ୰
ẘ⪅ࡀ⤫ྜኻㄪᵝࡢ≧ࢆ࿊ࡋࠊࡑࡢ≧ࡣᢠ⢭⚄⸆ࡢᢞࡼࡗ࡚ᢚไࡉࢀࡿࡇ࡞ࡀ
ሗ࿌ࡉࢀ࡚࠸ࡿ (4, 5)ࠋࡲࡓࠊLaruelle ࡽࡣࠊ⤫ྜኻㄪᝈ⪅ࡣᖖ⪅ẚ㍑ࡋ࡚ࢻࣃ࣑ࣥࡢᨺฟ
㔞ࡀ⣙ 2 ಸቑຍࡋ࡚࠸ࡿࡇࢆᐇドࡋ࡚࠸ࡿ (6)ࠋࡇࢀࡽࡢሗ࿌ࡽࠊ⌧ᅾ࡛ࡣࠕࢻࣃ࣑ࣥ௬
ㄝࠖࡀ⤫ྜኻㄪࡢែࢆㄝ᫂ࡍࡿ᭱ࡶ᭷ຊ࡞ㄝ⪃࠼ࡽࢀ࡚࠸ࡿࠋࠕࢻࣃ࣑ࣥ௬ㄝࠖࡣࠊ⬻ᖿ
㒊Ꮡᅾࡍࡿࢻࣃ࣑ࣥ⣔⚄⤒ࡢ⚄⤒᰾ࡽ⬻ࡢྛ㒊ᢞᑕࡍࡿࢻࣃ࣑ࣥ⤒㊰࠾ࡅࡿఏ㐩㞀ᐖ
ࡀ⤫ྜኻㄪࡢ≧ࢆច㉳ࡍࡿࡇࢆᥦၐࡋࡓㄝ࡛࠶ࡿࠋ࠼ࡤࠊ୰⬻⭡ഃ⿕㔝ࡽ㎶⦕⣔ࡢ
ഃᆘ᰾⮳ࡿࢻࣃ࣑ࣥ⤒㊰ (୰⬻㎶⦕⣔) ࠾ࡅࡿ D2 ཷᐜయࡢ㐣࡞่⃭ஹ㐍ࡣࠊዶࡸᗁぬ࡞
ࡢ㝧ᛶ≧ࢆច㉳ࡍࡿࡢᑐࡋࠊ୰⬻⭡ഃ⿕㔝ࡽ๓㢌ⴥ⓶㉁⮳ࡿࢻࣃ࣑ࣥ⤒㊰ (୰⬻⓶㉁
⣔) ࠾ࡅࡿ D1 ཷᐜయࡢ่⃭ᢚไࡣࠊពḧࡢపୗࡸឤࡢ㕌㯞ࠊ↓Ⅽ࡞ࡢ㝜ᛶ≧ࡸㄆ▱ᶵ⬟
㞀ᐖࢆច㉳ࡍࡿせᅉ⪃࠼ࡽࢀ࡚࠸ࡿࠋࢻࣃ࣑ࣥ௬ㄝຍ࠼࡚ࠊN-࣓ࢳࣝ-D-ࢫࣃࣛࢠࣥ㓟
(NMDA) ཷᐜయࡢ㠀➇ྜⓗᣕᢠ⸆࡛࠶ࡿࣇ࢙ࣥࢩࢡࣜࢪࣥࡸࢣࢱ࣑ࣥࡢ୰ẘ⪅ࡀ⤫ྜኻㄪᵝࡢ
≧ࢆ࿊ࡍࡿࡇࡸ (7)ࠊࢢࣝࢱ࣑ࣥ㓟ࡀ NMDA ཷᐜయ⤖ྜࡍࡿࡇࡽࠊࢻࣃ࣑ࣥ౫Ꮡࡋ
࡞࠸⚄⤒ఏ㐩ࡢㄪ⠇ࡼࡗ࡚⤫ྜኻㄪࡢ≧ࡀⓎࡍࡿࡇࢆᥦၐࡋࡓࠕࢢࣝࢱ࣑ࣥ㓟ఏ
㐩పୗ௬ㄝࠖ(8) ࡸࠕNMDA ཷᐜయᶵ⬟పୗ௬ㄝࠖ(9) ࡶ⤫ྜኻㄪࡢែࢆㄝ᫂ࡍࡿㄝ⪃࠼ࡽ
ࢀ࡚࠸ࡿࠋ
⤫ྜኻㄪࡢ⒪ࡣࠊᢠ⢭⚄⸆ࢆ⏝࠸ࡓ⸆≀⒪ἲࡀ୰ᚰ࡞ࡿࠋ᭱ࡶྂ࠸ᢠ⢭⚄⸆࡛࠶
ࡿࢡࣟࣝࣉ࣐ࣟࢪࣥࡀ 1951 ᖺྜᡂࡉࢀࠊ⩣ 1952 ᖺࡣࣥࢻ⺬ᮌ (Rauwolfia serpentina) ࡼࡾࣞ
ࢭࣝࣆࣥࡀࣝ࢝ࣟࢻࡋ࡚ᢳฟࡉࢀ࡚࠸ࡿ (10)ࠋࡑࡢᚋࠊࢡࣟࣝࣉ࣐ࣟࢪࣥࢆྵࡴࣇ࢙ࣀࢳ
ࢪࣥㄏᑟయࡣࢻࣃ࣑ࣥ D2 ཷᐜయᑐࡍࡿ㐽᩿స⏝ࠊࣞࢭࣝࣆࣥࡣࢻࣃ࣑ࣥࡸࣀࣝࢻࣞࢼࣜࣥ
࡞ࡢࣔࣀ࣑ࣥࢆࢩࢼࣉࢫᑠ⬊ෆ࡛ᯤῬࡉࡏࡿస⏝ࢆ♧ࡍࡇࡀ᫂ࡽ࡞ࡗࡓࠋࢡࣟࣝࣉࣟ
࣐ࢪࣥࡸࣞࢭࣝࣆࣥࡀ㛤Ⓨࡉࢀ࡚௨㝆ࠊࣈࢳࣟࣇ࢙ࣀࣥㄏᑟయ࡛࠶ࡿࣁࣟ࣌ࣜࢻ࣮ࣝ࡞ᙉຊ࡞
D2 ཷᐜయ㐽᩿స⏝ࢆ᭷ࡍࡿᐃᆺᢠ⢭⚄⸆ࡀከࡃ㛤Ⓨࡉࢀࠊ⮫ᗋ⌧ሙ࡛⏝ࡉࢀࡿ⮳ࡗࡓ
(Table 1) (11-13)ࠋᐃᆺᢠ⢭⚄⸆ࡢᙉຊ࡞ D2 ཷᐜయ㐽᩿స⏝ࡣࠊ⤫ྜኻㄪࡢ㝧ᛶ≧ᑐࡋ࡚
Ⰻዲ࡞ᨵၿຠᯝࢆࡶࡓࡽࡍࡢᑐࡋ࡚ࠊ㝜ᛶ≧ࡸㄆ▱ᶵ⬟㞀ᐖᑐࡍࡿᨵၿຠᯝࡣప࠸ࠋࡲ
ࡓࠊᐃᆺᢠ⢭⚄⸆ࡣ㯮㉁⥺᮲య⣔ࡸ₃ᩯୗᆶయ⣔࡞⤫ྜኻㄪࡢ≧㛵㐃ࡀప࠸ࢻࣃ࣑ࣥ
⤒㊰ࡢᶵ⬟ࡶᙉࡃᢚไࡍࡿࡇ࡛ࠊ㗹యእ㊰≧ࡸ㧗ࣉࣟࣛࢡࢳࣥ⾑࡞ࡢస⏝ࢆច㉳ࡍࡿ
1
ࡇࡽᚸᐜᛶࡀప࠸ࡇࡀၥ㢟ࡋ࡚ᣲࡆࡽࢀࡿࠋࡇࢀࡽࡢၥ㢟ࢆゎỴࡍࡿࡁࡗࡅ࡞ࡗࡓ
ࡢࡣࠊ1958 ᖺࡢࢡࣟࢨࣆࣥࡢྜᡂ࡛࠶ࡗࡓࠋࢡࣟࢨࣆࣥࡣᐃᆺᢠ⢭⚄⸆ࡢḞⅬ࡛࠶ࡗࡓ㗹యእ
㊰≧ࡢண㜵㝜ᛶ≧࣭ㄆ▱ᶵ⬟㞀ᐖ࡞ࡢ≧ᑐࡍࡿᨵၿస⏝ࢆ࠶ࡿ⛬ᗘඞ᭹ࡋࡓ⸆࡛
࠶ࡿࠋࡑࡢᚋࡢ⸆⌮Ꮫⓗ◊✲ࡼࡾࠊࢡࣟࢨࣆࣥࡣ D2 ཷᐜయࡢ㐽᩿స⏝ຍ࠼࡚ࠊࢭࣟࢺࢽࣥ 5HT2A ཷᐜయᑐࡍࡿ㐽᩿స⏝ࢆ᭷ࡋ࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡇࡢⓎぢࢆࡶࠊᐃᆺᢠ⢭
⚄⸆ྠ➼௨ୖࡢ㝧ᛶ≧ᨵၿస⏝ࢆ⥔ᣢࡋࠊ㗹యእ㊰≧ࡢண㜵㝜ᛶ≧࣭ㄆ▱ᶵ⬟㞀ᐖ
ᑐࡍࡿᨵၿస⏝ࢆ᭷ࡍࡿ⸆ࡢ㛤Ⓨࡀ⾜ࢃࢀࠊ1984 ᖺࢭࣟࢺࢽ࣭ࣥࢻࣃ࣑ࣥ㐽᩿⸆ (SDA) ࡛
࠶ࡿࣜࢫ࣌ࣜࢻࣥࡀྜᡂࡉࢀࡓࠋࡑࡢᚋࠊࢻࣃ࣑ࣥࡸࢭࣟࢺࢽࣥࡢཷᐜయ㐽᩿స⏝ຍ࠼࡚ࠊࣄ
ࢫࢱ࣑ࣥࡸࢻࣞࢼࣜࣥ࡞ࡢࣔࣀ࣑ࣥཷᐜయࠊ࣒ࢫ࢝ࣜࣥᛶࢭࢳࣝࢥࣜࣥཷᐜయᑐࡍࡿ
㐽᩿స⏝ࢆࡋࡓከඖཷᐜయᶆⓗᢠ⢭⚄⸆ (MARTA) ࡸࢻࣃ࣑ࣥཷᐜయ㒊ศసື⸆ (DPA) ࡞
ࡢ㠀ᐃᆺᢠ⢭⚄⸆ࡀ㛤Ⓨࡉࢀࠊ⌧ᅾ࡛ࡣ⤫ྜኻㄪࡢ➨୍㑅ᢥ⸆ࡋ࡚⏝ࡉࢀ࡚࠸ࡿ
(Table 1) ࠋ
Fig. 1. Structure of olanzapine.
࢜ࣛࣥࢨࣆࣥ (Fig. 1) ࡣ MARTA ࡢ୍ࡘ࡛࠶ࡾࠊࣔࣀ࣑ࣥཷᐜయࡸ࣒ࢫ࢝ࣜࣥᛶࢭࢳࣝࢥ
ࣜࣥཷᐜయࢆ㐽᩿ࡍࡿࡇ࡛㝧ᛶ≧㝜ᛶ≧ࠊㄆ▱㞀ᐖࡢᨵၿຠᯝࢆ♧ࡍ (Table 2) ࠋ2013 ᖺ
⾜ࢃࢀࡓฎ᪉ゎᯒㄪᰝ࡛ࠊ࢜ࣛࣥࢨࣆࣥࡣ᪥ᮏ࢝ࢼࢲ࡛᭱ࡶ⏝ࡉࢀ࡚࠸ࡿ㠀ᐃᆺᢠ⢭⚄
⸆࡛࠶ࡿሗ࿌ࡉࢀ࡚࠸ࡿ (14-16)ࠋ୍᪉ࠊ࢜ࣛࣥࢨࣆࣥࡢᢞࡣࡋࡤࡋࡤ㧗⾑⢾㉳ᅉࡍࡿ㔜⠜
࡞⢾ᒀᛶࢣࢺࢩࢻ࣮ࢩࢫࢆច㉳ࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ (17) ࠋᮏ㑥࡛ࡣࠊ࢜ࣛࣥࢨࣆࣥࢆ
᭹⏝ࡋࡓ 3753 ୰ 8 ࡛⢾ᒀᛶࢣࢺࢩࢻ࣮ࢩࢫࡀច㉳ࡉࢀࠊࡑࡢ࠺ࡕ 2 ࡢᝈ⪅ࡀṚஸࡋࡓ
ࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋࡇࡢែࢆཷࡅ࡚ࠊཌ⏕ປാ┬ࡣ 2002 ᖺ࢜ࣛࣥࢨࣆࣥࡢᢞࡼࡿ㔜
⠜࡞㧗⾑⢾ࡢࣜࢫࢡ㛵ࡍࡿ⥭ᛴᏳᛶሗࢆⓎ⾲ࡋ࡚࠸ࡿ (18)ࠋᢠ⢭⚄⸆ࡢ⏝ࡼࡿ⾑⢾
್␗ᖖࡢⓎࡘ࠸࡚ࡣࠊ⢾ᒀࡢ᪤ Ṕࢆ᭷ࡍࡿᝈ⪅ࢆ୰ᚰᐃᆺࠊ㠀ᐃᆺ㛵ಀ࡞ࡃሗ࿌ࡉ
ࢀ࡚࠸ࡿ (17, 19-22)ࠋࡋࡋ࡞ࡀࡽࠊࢡ࢚ࢳࣆࣥࡸ࢜ࣛࣥࢨࣆࣥ࡞ࡢ MARTA ࡣࡢᢠ⢭⚄
⸆ࡼࡾࡶ⾑⢾್␗ᖖࡢⓎ㢖ᗘࡀ㧗࠸ (23)ࠋ࢜ࣛࣥࢨࣆࣥࡼࡿ㧗⾑⢾ࡢⓎᶵᗎࡣᮍࡔ᫂ࡽ
࡛࡞࠸ࠋ㐣ཤࡢ◊✲࡛ࠊ࢜ࣛࣥࢨࣆࣥࡢᢞࡀ㣗ḧࡢஹ㐍ࡸయ㔜ቑຍࠊ⫧‶ࢆᘬࡁ㉳ࡇ
ࡋࠊࣥࢫࣜࣥᢠᛶࢆቑຍࡉࡏࡿࡇ࡛ 2 ᆺ⢾ᒀࢆច㉳ࡍࡿࡇࡀ♧၀ࡉࢀ࡚࠸ࡿ (24-28)ࠋ
ᐇ㝿ࠊṚஸࢆྵࡴ㔜⠜࡞㧗⾑⢾≧ࢆ࿊ࡋࡓᝈ⪅ࡢ⫼ᬒࡣ⫧‶ࡸ≧ࡢⓎ⌧┤๓ࡢ㐣㣗ࠊ⢾
2
Table 1. Classification of antipsychotic agents.
typical antipsychotic agent
phenothiazine derivatives
atypical antipsychotic agent
serotonin-dopamine antagonists (SDA)
Chlorpromazine
Risperidone
Levomepromazine
Perospirone
Perphenazine
Blonanserin
Fluphenazine
multi-acting receptor targeted antipsychotics
(MARTA)
Prochlorperazine
Clozapine
Trifluoperazine
Quetiapine
Propericiazine
Olanzapine
butyrophenone derivatives
Haloperidol
Asenapine
dopamine partial agonist
Spiperone
Aripiprazole
diphenylbutylpiperadine derivatives
Pimozide
benz amide derivatives
Sulpiride
Tiapride
Nemonapride
iminodibenzyl derivatives
Carpipramine
Clocapramine
Mosapramine
thiepine derivatives
Zotepine
ศࡢከ࠸Ύᾴ㣧ᩱỈࡢ㔞ᦤྲྀࡀሗ࿌ࡉࢀ࡚࠾ࡾࠊ㐣࡞⢾ࡢᦤྲྀࡼࡾࣥࢫࣜࣥࡢస⏝ࡸศ
Ἢࡢపୗࡀច㉳ࡉࢀࡓ᥎ ࡉࢀ࡚࠸ࡿࠋ࢜ࣛࣥࢨࣆࣥࡼࡿ㣗ḧࡢஹ㐍ࡸయ㔜ࡢቑຍࡢⓎᶵ
ᗎࡘ࠸࡚ࡣࠊ⬻ෆࡢ‶⭡୰ᯡ࡛⏕⌮ⓗ࡞㣗ḧࡢᢚไㄪ⠇㛵ࡍࡿࣄࢫࢱ࣑ࣥ H1 ཷᐜయࢆ࢜ࣛ
ࣥࢨࣆࣥࡀ㐽᩿ࡍࡿࡇ⏤᮶ࡍࡿ⪃࠼ࡽࢀ࡚࠸ࡿ (29) ࠋࡋࡋ࡞ࡀࡽࠊ࢜ࣛࣥࢨࣆࣥࡼࡾ
3
Table 2. Ki values of olanzapine against monoamine and acetylcholine muscarinic receptors (30).
receptor
Ki (nM)
receptor
Ki (nM)
D1
31
5-HT6
2.5
D2
11
5-HT7
104
D3
16
M1
1.9
D4
26
M2
18
D5
51
M3
25
5-HT1A
>1000
M4
13
5-HT1B
1355
M5
6
5-HT1D
800
α1
19
5-HT2A
2.5
α2
230
5-HT2B
11.8
β
>10000
5-HT2C
28.6
H1
7
5-HT3
57
GABAA
>10000
5-HT4
>1000
Benzodiazepine
>10000
ច㉳ࡉࢀࡿ㧗⾑⢾ࡣయ㔜ࡢቑຍࢆకࢃ࡞࠸ሗ࿌ࡶ࠶ࡿࡇࡽ (31-33)ࠊ㧗⾑⢾ࡢច㉳ࡣ
H1 ཷᐜయࡢ㐽᩿స⏝ࡼࡿ㣗ḧஹ㐍௨እࡢᶵᗎࡶ㛵ࡍࡿࡇࡀ♧၀ࡉࢀ࡚࠸ࡿࠋ࠼ࡤࠊOzasa
ࡽࡣ࢜ࣛࣥࢨࣆࣥࡢ᭚㟢ࡀࣥࢫࣜࣥࡢศἪࢆᢸ࠺⮅⮚ β ⣽⬊ࡢ࣏ࢺ࣮ࢩࢫࢆㄏᑟࡍࡿࡇࢆ
ሗ࿌ࡋ࡚࠸ࡿ (34)ࠋࡶࠊ࢜ࣛࣥࢨࣆࣥࡢฎ⨨ࡼࡾ⫢⮚࡛ࡢࢢࣝࢥ࣮ࢫ⏘⏕ࡀஹ㐍ࡉࢀࡿࡇ
ࡸ (35, 36)ࠊᮎᲈ࡛ࡢࢢࣝࢥ࣮ࢫࢺࣛࣥࢫ࣏࣮ࢱ࣮ࢆࡋࡓ⣽⬊ෆࡢ⢾ྲྀࡾ㎸ࡳࡀ㜼ᐖࡉࢀࡿ
ࡇ (37, 38)ࠊࢻࣞࢼࣜࣥࡢศἪஹ㐍ࢆࡋࡓ㧗⾑⢾ࡢㄏⓎ࡞ࡀሗ࿌ࡉࢀ࡚࠸ࡿ (39)ࠋ
ࣄࢺࣥࢫࣜࣥࡣࠊ21 ಶࡢ࣑ࣀ㓟ࡽ࡞ࡿ࣌ࣉࢳࢻ A 㙐 30 ಶࡢ࣑ࣀ㓟ࡽ࡞ࡿ࣌ࣉ
ࢳࢻ B 㙐ࡀࡘࡢ S-S ⤖ྜ࡛㐃⤖ࡋࡓ࡛࣍ࣝࣔࣥ࠶ࡿ (Fig. 2)ࠋ⮅ β ⣽⬊࠾࠸࡚ࠊࣥࢫࣜࣥࡢ
ࡶ࡞ࡿ 110 ಶࡢ࣑ࣀ㓟ࡽ࡞ࡿࣉࣞࣉࣟࣥࢫࣜࣥࡣ 24 ಶࡢ࣑ࣀ㓟ࡽ࡞ࡿࢩࢢࢼࣝ࣌
Fig. 2. Structure of human insulin.
4
ࣉࢳࢻࡀษ᩿ࡉࢀࠊࣇ࢛࣮ࣝࢹࣥࢢࢆཷࡅࡿࡇ࡛ࣉࣟࣥࢫࣜࣥ࡞ࡿࠋࣉࣟࣥࢫࣜࣥࡣ
ࡉࡽࣉࣟ࣍ࣝࣔࣥኚ㓝⣲ (PC2 ࠾ࡼࡧ PC3) (40, 41)࢝ࣝ࣎࢟ࢩ࣌ࣉࢳࢲ࣮ࢮ H (42) ࡢάᛶ
ࡼࡾࣉࣟࢭࢩࣥࢢࢆཷࡅ࡚ࠊࣥࢫࣜࣥ C ࣌ࣉࢳࢻ࡞ࡾࠊࡶศἪ㢛⢏୰㈓ⶶࡉࢀࡿࠋ
ࢢࣝࢥ࣮ࢫᏑᅾୗ࠾ࡅࡿࣥࢫࣜࣥศἪࡢ࣓࢝ࢽࢬ࣒ࡘ࠸࡚ࡣ Fig. 3 ࡢࡼ࠺࡞⤒㊰ࡀ⪃࠼ࡽࢀ
࡚࠸ࡿࠋࡍ࡞ࢃࡕࠊ⾑୰ࡢࢢࣝࢥ࣮ࢫࡀୖ᪼ࡍࡿࠊࢢࣝࢥ࣮ࢫ㍺㏦ᢸయ (GLUT) ࢆࡋࡓಁ㐍
ᣑᩓࡼࡾ β ⣽⬊ෆࢢࣝࢥ࣮ࢫࡀྲྀࡾ㎸ࡲࢀࡿ (43-45)ࠋྲྀࡾ㎸ࡲࢀࡓࢢࣝࢥ࣮ࢫࡣࠊゎ⢾⣔ࡢ
ᚊ㏿㓝⣲࡛࠶ࡿࢢࣝࢥ࢟ࢼ࣮ࢮࡼࡾࢢࣝࢥ࣮ࢫ 6-ࣜࣥ㓟 (G6P) ௦ㅰࡉࢀࡿ (46)ࠋ⏕ᡂࡋࡓ
G6P ࡣゎ⢾⣔࡛ࡉࡽ௦ㅰࡉࢀࡿࡀࠊࡇࡢ㐣⛬࠾࠸࡚ ATP ࡀ⏘⏕ࡉࢀࠊ⣽⬊ෆ ATP/ADP ẚࡀ
ୖ᪼ࡍࡿࡇ࡛ ATP ឤཷᛶ K+ (KATP) ࢳࣕࢿࣝࡀ㛢ཱྀࡋࠊ⣽⬊⭷ࡀ⬺ศᴟࡍࡿࠋKATP ࢳࣕࢿࣝ㛢
ཱྀࡼࡿ⣽⬊⭷ࡢ⬺ศᴟࡼࡾ⭷㟁ࡀ L ᆺ㟁౫Ꮡᛶ Ca2+ࢳࣕࢿࣝ (L-type voltage-dependent
Ca2+ channel, L-type VDCC) ࡢ㜈⭷㟁ࡲ࡛㐩ࡍࡿ L-type VDCC ࡀάᛶࡉࢀࠊ⣽⬊እࡢ Ca2+ࡀ
ὶධࡍࡿࠋࡑࡢᚋࠊᑠ⬊యࡽࡢ Ca2+ᨺฟࡀ⥆ࡁࠊCa2+⃰ᗘࡢୖ᪼ࡼࡾࣥࢫࣜࣥศἪ㢛⢏ࡣ
⣽⬊⭷ࢺࣛࣥࢫࣟࢣ࣮ࢺࡋࠊࣥࢫࣜࣥࡣ࢚࢟ࢯࢧࢺ࣮ࢩࢫࡼࡾ㛤ཱྀศἪࡉࢀࡿ (47)ࠋ
Fig. 3. Mechanism of physiological insulin secretion from pancreatic β-cell.
5
ࢻࣞࢼࣜࣥࡸࢭࢳࣝࢥࣜࣥ࡞ࡢ⚄⤒ఏ㐩≀㉁ࠊࢢࣝ࢝ࢦࣥᵝ࣌ࣉࢳࢻ (GLP-1) ࡞ࡢ
ࣥࢡࣞࢳࣥࡣ࢝ࣝࢩ࣒࢘ࢩࢢࢼࣝࢆࡋࡓࣥࢫࣜࣥศἪࢆஹ㐍ࡉࡏࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ
(48)ࠋ࠼ࡤࠊࢻࣞࢼࣜࣥࡸࣀࣝࢻࣞࢼࣜࣥࡣ⮅ β ⣽⬊ࡢࢻࣞࢼࣜࣥ α2 ཷᐜయࢆ่⃭ࡍࡿ
ࡇ࡛ࣥࢫࣜࣥศἪࢆᢚไࡋ (49, 50)ࠊβ2 ཷᐜయࢆ่⃭ࡍࡿࡇ࡛ศἪࢆಁ㐍ࡍࡿ (51)ࠋࡲࡓࠊ
ࢭࢳࣝࢥࣜࣥࡣ࣒ࢫ࢝ࣜࣥᛶࢭࢳࣝࢥࣜࣥ M3 ཷᐜయࢆ่⃭ࡍࡿࡇ࡛ࣥࢫࣜࣥศἪࢆஹ㐍
ࡍࡿ (52)ࠋࡉࡽࠊGLP-1 ࡸᾘ⟶ᢚไ࣌ࣉࢳࢻ (GIP) ࡣࢢࣝࢥ࣮ࢫ่⃭ୗ࡛ྛཷᐜయࢆ่⃭ࡍࡿ
ࡇࡼࡾࣥࢫࣜࣥࡢศἪࢆಁ㐍ࡍࡿ (53, 54)ࠋࡇࢀࡽࡢሗ࿌ࡣࠊ⮅ β ⣽⬊ࡽࡢࣥࢫࣜࣥศ
Ἢࡢㄪ⠇⮅ β ⣽⬊Ⓨ⌧ࡍࡿࣔࣀ࣑ࣥࡸࢭࢳࣝࢥࣜࣥࠊࣥࢡࣞࢳࣥ࡞ࡢཷᐜయࡀ┤᥋
㛵ࡍࡿࡇࢆ♧၀ࡍࡿࡶࡢ࡛࠶ࡿࠋ࢜ࣛࣥࢨࣆࣥࡣ✀ࠎࡢࣔࣀ࣑ࣥཷᐜయᑐࡋ࡚ᣕᢠస⏝
ࢆ♧ࡍࡇࡽࠊ࢜ࣛࣥࢨࣆࣥࡀ⮅ β ⣽⬊࠾ࡅࡿࣔࣀ࣑ࣥཷᐜయࡢ㐽᩿స⏝ࢆࡍࡿࡇ࡛
ࣥࢫࣜࣥศἪࢆᢚไࡋࠊ㧗⾑⢾≧ࢆច㉳ࡍࡿྍ⬟ᛶࡀ᥎ᐹࡉࢀࡿࠋࡋࡋ࡞ࡀࡽࠊ⮅ β ⣽⬊
ࡽࡢࣥࢫࣜࣥศἪ࠾ࡅࡿࢻࣞࢼࣜࣥཷᐜయ௨እࡢࣔࣀ࣑ࣥཷᐜయࡢ㛵ࡘ࠸࡚ࡣ᫂
ࡽ࡛࡞࠸ࠋᮏ◊✲࡛ࡣࠊࣁ࣒ࢫࢱ࣮⏤᮶⮅ β ⣽⬊ᰴ࡛࠶ࡿ HIT-T15 ⣽⬊ࢆ⏝࠸࡚ࠊ⮅ β ⣽⬊
ࡽࡢࣥࢫࣜࣥศἪ࠾ࡅࡿࢻࣃ࣑ࣥࠊࢭࣟࢺࢽࣥ࠾ࡼࡧࣄࢫࢱ࣑ࣥཷᐜయࡢ㛵ࢆゎ᫂ࡍࡿࡇ
ࢆ┠ⓗࡋࡓࠋ
6
ᮏㄽ
1. ࣁ࣒ࢫࢱ࣮⏤᮶⮅ ș ⣽⬊ᰴ (HIT-T15) ⣽⬊࠾ࡅࡿྛࣔࣀ࣑ࣥཷᐜయࡢⓎ⌧ゎᯒ
1-1. ┠ⓗ
ᮏ◊✲࡛⏝ࡋࡓ HIT-T15 ⣽⬊ࡣࠊSyrian hamster (Mesocricetus auratus) ࡢ⮅ࣛࣥࢤࣝࣁࣥࢫ
ᓥ⣽⬊ࢆ simian virus 40 ࡛ࢺࣛࣥࢫࣇ࢛࣮࣒ࡍࡿࡇࡼࡾᶞ❧ࡋࡓࣥࢫࣜࣀ࣮࣐⣽⬊ᰴ࡛࠶ࡿ
(55)ࠋHIT ⣽⬊ࡣ࠸ࡃࡘࡢࢧࣈࢡ࣮ࣟࣥࡀᏑᅾࡍࡿࡀࠊࡑࡢ୰࡛ࡶ HIT-T15 ⣽⬊ࡣࣥࢫࣜࣥ
ྵ᭷㔞ࡀከࡃࠊࢢࣝࢥ࣮ࢫ⃰ᗘ౫Ꮡⓗ࡞ࣥࢫࣜࣥศἪస⏝ࡔࡅ࡛࡞ࡃࠊࢢࣝ࢝ࢦࣥࡸࢫࣝ࣍ࢽ
ࣝᒀ⣲⸆ࡼࡿࣥࢫࣜࣥศἪࡢஹ㐍ࡸࢯ࣐ࢺࢫࢱࢳࣥࡼࡿศἪࡢᢚไࡀሗ࿌ࡉࢀ࡚࠸ࡿ (5658)ࠋࡲࡓࠊṇᖖࡢ⮅ᓥྠᵝ HIT-T15 ⣽⬊ෆࡢ cAMP ⃰ᗘ⣽⬊እࡢ K+⃰ᗘࡢቑຍࡣࣥࢫࣜ
ࣥࡢศἪࢆஹ㐍ࡍࡿ (56)ࠋࡇࡢࡼ࠺࡞⌮⏤ࡽࠊHIT-T15 ⣽⬊ࡣከࡃࡢࣥࢫࣜࣥศἪᐇ㦂⏝࠸
ࡽࢀ࡚࠸ࡿࠋ
⮅ᓥࡸ⮅ β ⣽⬊࠾ࡅࡿࣔࣀ࣑ࣥཷᐜయࡢⓎ⌧ࡘ࠸࡚ࡣࠊ࠸ࡃࡘࡢሗ࿌ࡀ࠶ࡿࠋ࠼
ࡤࠊࣛࢵࢺ⮅ᓥࢻࣃ࣑ࣥ D2, D3 ࠾ࡼࡧ D4 ཷᐜయࡀⓎ⌧ࡋ࡚࠸ࡿࡇ (59-61)ࠊ࣐࢘ࢫ⮅ᓥ D2,
D3 ཷᐜయ (61) ࠾ࡼࡧࢭࣟࢺࢽࣥ 5-HT2C ཷᐜయࡀⓎ⌧ࡋ࡚࠸ࡿࡇ (62)ࠊࣛࢵࢺ⏤᮶⮅ β ⣽⬊ᰴ࡛
࠶ࡿ INS-1 ⣽⬊ D2, D3 ཷᐜయ (59, 63) ࠾ࡼࡧ 5-HT2B ཷᐜయࡀⓎ⌧ࡋ࡚࠸ࡿࡇ (64)ࠊ࣐࢘ࢫ⏤᮶
⮅ β ⣽⬊ᰴ࡛࠶ࡿ MIN6 ⣽⬊ D2 ཷᐜయ (61)ࠊ5-HT2B ཷᐜయࡀⓎ⌧ࡋ࡚࠸ࡿࡇ (65) ࡀሗ࿌ࡉࢀ
࡚࠸ࡿࠋ୍᪉ࠊᮏ◊✲࡛⏝ࡋࡓ HIT-T15 ⣽⬊࠾࠸࡚ࡣࠊࡇࢀࡽࡢࣔࣀ࣑ࣥཷᐜయࡢⓎ⌧
㛵ࡍࡿሗ࿌ࡣ࡞࠸ࠋࡑࡇ࡛ࠊ࢜ࣛࣥࢨࣆࣥࡢせ࡞స⏝㒊࡛࠶ࡿࣔࣀ࣑ࣥཷᐜయࡢ mRNA Ⓨ
⌧ࡀ HIT-T15 ⣽⬊࡛☜ㄆ࡛ࡁࡿྰࢆ RT-PCR ἲ࡛☜ㄆࡋࡓࠋࡉࡽࠊ≉␗ⓗ࡞ᢠయࡀධᡭ࡛
ࡁࡓࢻࣃ࣑ࣥ D2 ࠾ࡼࡧࢭࣟࢺࢽࣥ 5-HT2A ཷᐜయ㛵ࡋ࡚ࡣࠊࢱࣥࣃࢡ㉁ࡢⓎ⌧ࢆ western blot ἲ
࡛ゎᯒࡋࡓᇭ
1-2. ⤖ᯝ
ྛࣔࣀ࣑ࣥཷᐜయᑐࡍࡿ≉␗ⓗ࡞ࣉ࣐࣮ࣛࢆタィࡋࠊRT-PCR ࡛ cDNA ࢆቑᖜࡋࡓᚋࠊ
2%࣮࢞ࣟࢫࢤ࡛ࣝ㟁ẼὋືࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊD2 ཷᐜయ࠾ࡼࡧࣄࢫࢱ࣑ࣥ H1 ཷᐜయࡘ࠸
࡚┠ⓗ㛗ࡢࣂࣥࢻࡀ᳨ฟࡉࢀࡓࡀࠊࡑࡢࡢཷᐜయࡢⓎ⌧ࡣ☜ㄆ࡛ࡁ࡞ࡗࡓࠋࡑࡇ࡛ࠊⓎ⌧ࡀ
☜ㄆ࡛ࡁ࡞ࡗࡓ D3, D4 ཷᐜయࡸ 5-HT2A, 5-HT2B, 5-HT2C ࠾ࡼࡧ 5-HT6 ཷᐜయࠊH2 ཷᐜయᑐࡋ࡚
2 ẁ㝵 PCR ࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊ࡚ࡢཷᐜయࡘ࠸࡚ mRNA ࡢⓎ⌧ࡀ☜ㄆ࡛ࡁࡓ (Fig. 4)ࠋࡉࡽ
ࠊD2 ࠾ࡼࡧ 5-HT2A ཷᐜయࢱࣥࣃࢡ㉁ࡢⓎ⌧ࢆ western blot ἲ࡛ゎᯒࡋࡓ⤖ᯝࠊ࣌ࣉࢳࢻ-N-ࢢࣝ
ࢥࢩࢲ࣮ࢮ (PNGase) ฎ⌮ᚋࡢ࣍ࣔࢪࢿ࣮ࢺ୰࡛ࡑࢀࡒࢀࡢཷᐜయࡢศᏊ㔞┦ᙜࡍࡿ⣙ 60 kDa
࠾ࡼࡧ⣙ 48 kDa ࣂࣥࢻࡀ☜ㄆ࡛ࡁࡓ (Fig. 5)ࠋ
7
Fig. 4. ...