!"#$%&
!
"#$%&'( )% *+,-./0123&
4'56789$%5:5'; <<( 1=&,-
">?@ ABCD( ) , -E<5'F#?G; ?F>
H I( /0,-
&JK-L MNO<(
">&PQ$%RS( &!"$%
#$ TUVWXY;
Z[\]F^7_%?&`ab '(( c)
&*d)+O<5,'%( efg\higjk
\( [lUm( g\hig\@0
"-.&no%5'; /pFZ[\]&n o%5' q0&1<( Z[\]
krst,S uh 2v3 Y<( ) 2v3 &`ab '(( c)&
wTUVWX YxB;
l[\yzg10{&4<5Z[\]UR100 |}
~W\200 mLt,S24do? Q<
5678( Q<RF 9&A:
;(Bond Elut C18&A<=9( zS(0
,\eO<R; Z[\] &no%>0
( [lUm( efigjk\( g\higj
\( 4A'?$%2v3n590 0 4O<( krs 4A\e&+
'5HPLC-ESI-MS-MS+&A@AxR; @AB+
&+'5FC/p"D<RB+1, 2),<5(
3 UeE&>0F( jefU mF([lUmFF)%%ESI negative mode?(
Xy|\F4A) Oefg\higjk\
FFESI positive mode?@AxR;
G HF$5'R7"(
!"&J¡¢I&£;
Vanillic acidF¤.¥' !"J£<R"( ¦ ¦O FZ[\]&Ano%5'efg\higjk\
W 2v3O<5. &!"$%5'§-"
£¨$%R3);oRIsovanillic acid,m-Coumaric acid(4 Am-Hydroxyphenylacetic acidFkrs &F©
9$%ª( &«5©9$%R. ?( ¦%.Z[
\] q0 2v3O<5 !"$
%5'¦O"¬%R; oR/0[lUmKL .!"JF>'. &!"$%¦O"8%(
$&Catechin4AEpicatechin&+'5F( )%
2v3?G3῎-4A4῎-O-. &!"
$%5'¦O8<R4); ®B( Z[\]&-.
&no%5'efg\higjk\( M&A-type Pro- anthocyanidinF&NO¯ P° QRS±² ,"T%54D5)( Z[\]"&`ab '(UFc)w¢ K-O$%5'"( A-type Proanthocyanidin "V oo³´S( &!"$
%&?) w¢E<5'FµW?G; )¦?
u h & 4 ¶ Procyanidin A-2 ! "& + ' 5 HPLC-ESI-MS-MS&AD©YxR¡¢( 'ª%
\e &.8?:R"( ) 2v3O¬
%·&<X¸Z©9¦O"?
:( Procyanidin A-2F>?$% &!"
$%¦O£¨YR ¹;
¡¢krs &F8%'º+
" &!"$%5'¦O"OD(
Z[\] &`ab c)( '(&J<5F( kr s q0.$¦O">2v3.) w¢&
' uh&4¶Z[\]krst,S !"(n»10)
Compound
Urinary excretion (mM/24 hr)
(min-max)
Excretion recovery
(¼)
Vanillic acid 84.0 (2.3῍703.6) 89.3
Isovanillic acid 10.0 (0.71῍42.6) ῌa) Protocatechuic acid 15.7 (0.26῍74.9) 6.0 m-Coumaric acid 9.7 (0.63῍42.2) ῌ p-Coumaric acid 17.4 (2.37῍70.0) 8.1 m-Hydroxyphenylacetic acid 9.3 (0῍85.6) ῌ Chlorogenic acid 0.14 (0.01῍0.42) 0.72
Isorhamnetin 1.0 (0.17῍4.9) 0.69
Quercetin 0.58 (0.11῍2.8) 0.80
Quercitrin 0.06 (0῍0.15) 0.21
Quercetin 3-O-glucoside 0.08 (0.01῍0.40) 0.11 Quercetin 3-O-arabinoside 0.02 (0῍0.16) 0.04 Myricetin 3-O-galactoside 0.02 (0῍0.05) 0.03
a) krs ?F©9
! "
#$!%&'
( )*!+,-'. )*/0!1.
234567)*/89:;<=2- 1) H. Ito, M.P. Gonthier, C. Manach, C. Morand, L. Mennen,
C. Remesy, and A. Scalbert, BioFactors, 22, 241῎243 (2004).
2) H. Ito, M.P. Gonthier, C. Manach, C. Morand, L. Mennen, C. Remesy, and A. Scalbert, Br. J. Nutr., 94, in press (2005).
3) L. Y. Rios, M. P. Gonthier,, C. Remesy, I. Mila, C. Lapi- erre, S. A. Lazarus, G. Williamson, and A. Scalbert,Am.
J. Clin. Nutr.,77, 912῎918 (2003).
4) >? @ABC 0 D EF? G HI 56782005 !8 JK"L 5) L. Y. Foo, Y. Lu, A. B. Howell, and N. Vorsa,J. Nat. Prod.,
63, 1225῎1228 (2000).
ῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌ
#M77$ N% O
P&Caenorhabditis elegansQ #R'! <STU VWXY #RZ[ \6]()*&!^_
`+- a )*&^_QTGF-bbcdinsuline fgchij klinsulinefgQP
& _&m,!ij-a"-n P&Q`+
./opq JrQ`+./0sL !t_T12
#R'!!-u 67vw3 x4! o pqQTGF-b t_T12!5y P&!`+z a opq 6{Q20 |Y7}n-n ~ 8Q9: 22'h
;nQ `+./opq r<!:n}-&
!u 2005 2 )*f
c.`+./opq Jdaumone"L 8= > ;nQ )* ?ῌ`+
./$!@-0sA6{-a" ῍`+./
opqBC0_&m,!Dz$!@-a
"! aaQ opqBC0insulin efg!_&m,!Dza"=
-
%#N2 <EF!, P&bcdU'G
! $Hof J
IL !, 1/100JK24 opqBC
0" 2 opq#t9Ldaf-22
MN ¡!, O¢£" #REP(NGM:
nematode growth medium) ~¤ BC0!
10¥JKQk `+RJ)*&^_RL!S"
a¦ %#TU BC0QV90¥ `+R!
O opq#t9LTU BC0Q WY)*&!^_X}h
m,SQ?X 6{S!Y§-u #R EP5-fluoro-2῏-deoxyuridine!QkZ[#!5 y <¨NGM%#TU BC0!5¥' Q10¥JKQk4\*&!25©EFm,!S ~ ª« ]¬,Q~¤V1720 'h 2BC0!klXt ]¬m,QV13 'h a a"}n %#TU BC0Q4!
®6_&m,!Dza":n}"Xh
! q¯°±²³3 ´Procyanidin A-2 pµ6< HPLC-ESI-MS-MS1¶b·
e q f ¸ ¹ q o f º (MRM mode with positive ionization)
a)(m/z) parent ion/(m/z) product ion
! P& `+Tm,!ij-»=^_¼
῍ 6 ῍
! "#
$%&"'&(
)*
insulin
daf-16+,$-./01234,5.26
78 9 :$insulin;<=> ?@AB !
"DAF-2C9#DPI 3-kinase :E F
&G$%%& 9H insulinI&J'%&KL%
( :H$ G)*6
$insulinI&M%
&NA
OP insulin+, Q+,-R%
& K.%&S T/01U2Q U2%& 3V
WP4XY(6Z[5\V'6 @]E6^
_`XY78a9Db:6'%
1) c de 2005;<=46^_`a>a?@
ῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌ !"#$3-%&'()
*+,-./01234.567
fA>`6`B6gh`h CDij
EFkl%&mHn&op;2qr;$
GHIsto.u v_`wJ678 6 xKLM\yNOPj zk {|}2<o~ . I (EC2.7.8.13 Phospho-MurNAc-pentapeptide translocaseE nBt ..HdQ-R%& 'H XYH op;2q r;A7 R 3-}><>>
_ @r|(IV)E D$ST2 U (6 mk(6F8mkU2V6
& @W1E1) 9H r|(IV) U2I%&
lXYR Zr|(III) _ Hn&op;2qr;C-(III) n&$M-(III)2)o
26K +,[ 3-}><>>
\:]%U2+^_` 96 9 U2_`op;2qr;\:
](/Ia6 b/@cE 2.%&d]6XY3 @W 2E
(1) /893-%&'()*:+,-.;
<=>?
ῌv_`ee op;2qr;M-(III)K 0.1 Mf< pH 3pH 10 ^gh 37 24i
e7*&r|(III)r|(IV)
HPLCF8j pH 8 '8 pH
10H$k61¡ j pH 7!¢H
$D_$ lWm
pH 10 @£¤῎¥_~o¤¦῎n_=o¤¦E
gK 9 f< §F&mj
0.2 M!Hr|(III)r|(IV)
100¡ !F8 opq¨r
/ ''H 3-}><>> \:]
%v_`]r©.3) Oª (F8 str© F3-}><>> op;2q;r r|(II)(IV)« $ 9 lX u¬ sv pH_F&.
®¯°/ ±w2x
῍_`eev_`]+,[ 3-}><>>
\:]%r©. H l
\:]cU2 ²³ 9^ yz_`
@¤; ¤q ´ µ¶ ·¸¹ ¤º¶ »uo 1 >1E Kop;2qr;e
HPLCH+{%&Hr|(III) r|(IV) U26 r©$ 40 mMo
g(pH 8)h500mg ^_`K37
24i e 9 ( ¤º¶1`h 40¡|A&U2 9H O P (/ }¼$ ¤º¶1 `
K&~63)
@1 op;2qr; wJ
(2) 3-
ῌ
3-
!"#$%
C-(III)&'()*(+,-.#$/0
""#1 2% &34 0 " 5 6% Biotinamidohexanoic acid N-hydroxy- succinimide ester "4-Dimethylaminopyridine 7
$89:"0;% < =>?
HPLC@ABC71D+EF6G1 H0
@% HD+E#$IJ(ESI(K)) 9:"0;% (LMN 1269.61759% N 1269.61789)% O(C58H93N8O21S)F6G%
F@P"#1 QRST#
@% C-(III)UVWXV
&Y&Z[+U\]^_% `ab% -cVd
^_0"@\]eE9f!@71
"#$%&gh'ijk($)*g56+,- l]$ mnX[+op q*#% HG7$%
.r/Ws,)t.jk#01ujv 2Jw3,n[Y+2% vx34y56 7z7@{>% H=fj 8|}=>% QRj
~9:h':;"\!71
R%|}/# <=P# mp>z|
}?@6AB#1
1) K. Kimura and T. D. H. Bugg:Nat. Prod. Rep.(Review), 20, 252῎273 (2003).
2) S. Kagami, Y. Esumi, M. Nakakoshi, M. Yoshihama and K. Kimura:J. Antibiot.,56, 552῎556 (2003).
3) CD %EFG% CH% IJK%L MN% O P%IQ%R%STN2003UVW>
zXYZ% p. 272.
ῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌ
2,6-2 !"#$%&'()*+
,-./012
[\XzXz>z|} ]^M_
U+e+a (O2ῌ)2% S `@E[
'+Yjka=:$# ^G% ¡¢hbcd Jjk#e£fYg¤¥j$¦F`§¨=>
©h# ^G"ªihj% ^6«¬!j F"®¯6G$71 0kj°l56% [\Xz>z
m8#0z#nSz|}o@2O2ῌ # pq]$r±F9²G$P1 ³% s|}o@
2&', ($Cassia spectabilis=>O2ῌ
#pq2,6-2t´DµY&,[ spec-
tamine A (1), spectamine B (2), iso-6-cassine (3)uv#
1 0G6]$f¶s·A!specta- mine A (1)225mmol@E['+Y=># ^G O2ῌ46.7¸pq#12,6-2t´DµY&,[
2kj#nSFc¹^G$7F% Sp qº»2|}F2¼$@{1|}@2S 32 &'()*(+,-.=3-
῍ 8 ῍
2,6-2 !"#$%&'()*
+,-./0012 3Spectamine A (1)4)56
@ABCDEFGH<@ABC$6IJ2 KL MN" O!$PJQR*S.!$DT NaN3PJU4SVWT.XYZ'[!$\]
7I^/0_[`2
a4)bEFTp!"cde fgH,g hi(jDEF[k.22,6-cisg j4) Tl!"!"7"7XLPdCl2mnPJ./
0[op(!qrsthiu[EF_vw/0x8 1= (y)-cassine4)FP1(Fig. 1).
z{T4)_|}0K~.2,6-transg)K.
1(Fig. 2).
= !"$8PJmnPd(PPh3)4
PJU45 : 2[w2,6-transg_L~
2 PdVNiPtmnPJU4T
E1<):_L~(X2
!"!"7"#"[17
IgPJ<,gP1<2,6-transg4)
_6 ",gEF_v¡1 (X(Fig. 3).
Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 2 Pddppp
9Scheme 2
spectamine A (1)!" #$%&' ()
spectamine A (1) 2,6-2*+,-./01 2345678 9:;<
=>?@6A67 BCD E"67 F2GHIJK3LM NO
PQ R<D ST
%?O!UV")WXY"Z[E)" \]^#_
PQUV`$a=bc%)dE
ῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌ
&'(Q(Qe#QUVf ghEN
1. LC/MS
i-2jkl5A (polycavernosideA, PA)9O1991) m0I)" Gracilaria edulis *+,no -p)O qr./O @s0)"1tu3-5
(Fig. 1)1) m0Ivw9132x)
33y4)" z{k-*+,569O * )7)|89:,P O y4;}
<!REO ~ =R4wz{
k-+,=O 33y="
-+/'>?La Union "2*
8O 2z{k-G. edulisk-
Acanthophora specifera NO 2002)῎2003)3
*+,)O 362x)O 83y4)"
PAno-p)"";O @
A)B0)"2)Fig. 1 CO i-2jkl5 9D=E$ 3) F,|8"t
"), O LC-DGHUV( (LC- DAD) (Mightysil RP-18, 0.215 cm, MeCN῎H2O 4 : 1, v/
v, 26, 0.2 ml/min) )"O i-2jkl5
56 I-/ (E, E, E) (lmax259, 269,
280 nm) +LuO JKPAB¡¢£¤
" O ¥B¡LCLwhO TSQ700
"LC / ESI-MS (MNaῌ( SIM¦L5)) O i-2jkl50K-l0§l/JK tu3tm¨IM©)"ª«O PA¢£¤
N +Lu" O i-2jkl5 MNaῌO4z/)"MS/MSJP9O 11¤O
10m-¬/¤ (Fig. 1 (a)῎(d))O
LC/MS/MS SRM¦L5)PA®QB
0)" LC-DADLC/MS (SIM)ª«PA| 8¯°SR9O S80 nmol/kg±|8T0)"
2. !"#$%&4)
2002)12²2<-+/*+,£
3<? " G. edulis (40 g) 0.1 LU5 VO ³V´µW/(3000 rpm 15 min))
"XYO |8Z[-)" Z[-V¶"|8
³·OacetoneMeOH¸\)"]¹?O CHCl3῎40ºMeOH)O CHCl3t (10 g, ddYO^Oi.p.)_`)"24£¤a1bt
y4»,R1 MU O Z[-93 MUO V¶"|892 MU,(" Z[- CHCl3O Silica gel \Wakogel C-100, 1 ml, CHCl3῎ MeOH, 9 : 1c^ ¯ Silica gel ODS Q3 \d¼
Scheme 2
῍10῍
1 ml, 100MeOH LC-DAD LC/MS (SIM), LC/MS/MS (MEM)
PA !
"# 2$%&'$() *+,-.
/ %012&345
Paquette67./ 85PA9:;<PP5 (Fig. 1)= PA>?@ABCD&>ABEBAB (E,E)FG85H9(I JKLMN1OP&
PAQ(LD99, ddY, mice, i.p. 0.4῍0.8 mg/kg)( 15)"PP5R STMN1UVW DMSOX YZ[\]0.5PP5^?P_`
PabcT(K562)d0 JKLefghT(Neu-
ro-2a)ijN1 Tk12mMlmHn
24῍48opqrs sN1tDAPI uvN1 wxJyBz{& !85 | Neu- ro-2a";<12mMij} 5op}caspase- 3/7~PN1 +&n85 | H8 5DNAuvN1TUNEL PP5ij}24op
&vuv85T& !85 8/
ij}22op}T./DNA
N1 H85DNA& !85 *+0IPP5=Neuro-2a12mM?L
N1"&n85
Z} MR|Y
H ¡¢R|N
1) Mari Yotsu-Yamashita, Robert L. Haddock, and Take- shi Yasumoto,J. Am. Chem. Soc.,1993, 115, 1147῍1148.
2) Mari Yotsu-Yamashita, Takeshi Yasumoto, Shunichi Yamada, Fe Farida A. Bajarias, Mirriam A. Formelozsa, Marc Lawrence Romero, and Yasuo Fukuyo,Chem. Res.
Toxicol.,2004, 17, 1265῍1271.
3) Mari Yotsu-Yamashita, Tetsuya Seki, Valerile J. Paul, Hideo Naoki, and Takeshi Yasumoto,Tetrahedron Lett., 1995, 36, 5563῍5566.
4) £¤|I ¥¦§ ¨©ª «¬ ® ¯£¨
°¦ Fe F. A. Bajaria, Mirriam A. Formelosza ±²
³ ´46µ¶·¸H9¹º»¼{ p. 671῍676 (2004).
5) Louis Barriault, Serge L. Boulet, Kenshu Fujiwara, Akio Murai, Leo A. Paquette and Mari Yotsu-Yamashita, Bioorg. & Med. Chem. Lett.,1999, 9, 2069῍2072.
Fig. 1 The structures of polycavernoside congeners and their ESI-MS/MS fragmentation patterns (aῌd)