Leukotriene A4 hydrolase 阻害薬の抗掻痒作用に関する研究

(1)

Title

Leukotriene A4 hydrolase 阻害薬の抗掻痒作用に関する研究(

_{本文（FULLTEXT) )}

Author(s)

辻, 文雄

Report No.(Doctoral

Degree)

博士(薬科学) 連創博甲第4号

Issue Date

2010-09-30

Type

博士論文

Version

publisher

URL

http://hdl.handle.net/20.500.12099/35015

※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。

(2)

Leukotriene

A4 hydrolase

_FRS#e)

bt3S*1fjB

8=

Bg+

8 #3#9E

i /IIP> :_A( ..t i_ i/..∼

2010*9B

P*Ji

A

_{jB%* JB3?B;3*%Br]#}

Bi

#.F##q5ES+

B;i.F##3i**

i+

X#

(3)

Leukotriene A4 hydrolase

_{FRS#e)bt35*fFjn=}

_Bg+

8

_qge

I II()( rll'i_'[l--J[::-.; ];-)-].:_L1:I,-:

:-(i-

1

It!T]-::lJi-2

fn-･'l't7;3 ItlT)-･'tTi-4

I.1i

:Ti-.5

fill-:tTli-6 [[ri ::Til7 )'t;[l-･'Tll8

Ilo-･ . . 3

]iJi.JT-_tiff.1..'.(j1.,i-.r"iiETu) _{;:[L_1?n'J..l1)i} . . .

L1

･l/:n'iff,.1tiLlui:)'T::(})1e1:Lli 1}I _-/:ll . . .

i

I:7₁ = _{f, I)}=t= i A1 )JL]-/I('J))'['J,'I!-L.i.1:=*i:.'L{)/- Lit

tv I,._L:L lil-ui4rI':-_tiff.1;.(:iii-(})_{I.[7'J.EIJI:_}

2 _{/)(})ft;BT'JtE,i-i-i-;'(:,)}[I ₁ = _{T, I)}=L i AI I)Il_{/I(I)))'=].Tlli.YJ-:ji:.}

-l1,1;ll--t> I,lJJ[1/I('J)I.'J'l!n-:Illi =tt(J)} -))I- 7:i,

7' _-I/_/;/+ _{//-i;1l-:Ill._L}_=i-J)} -I)I 7:i,

l!.Tt

((--,J)

t=I₁ =7 I, T)=t= i A i))H7J('J)I.i,'T'L[Ti.i:.I-;J,.i:-I;II:.i;-::;;.i

･1-il1-)rJ)i:(j)Fit[')(J

:JlilL1:I.-:-:1J'::I-():ilT=I₁ = _{I, T)}=T= i A1 ))H7J(;)I.'J'rJ["li:---:)..i:-I;tlJ[;I-:,].i3SA6{)41 ,/))l{_{)..F)I:JliJ)iJ.'['f[Hl-:}_lTl

.:i-(.I-1

1q!T1.

TT-)F'j4=iu)Flrr1() ･iJfl-2

[jTJ-:Ti-3

1't!T]-･'Ti-4

[!lti

:lTti-fT) [[rJ-･'Ti-6 [.i)-･'lTi-7 [n.

tj₁ = _{I, T)}=T= i _^J)I]-/I(_{'J)(.'J,'r'tTi.Y:-:L:-I:_uJ.i-,.::Flii}SAG.,-)41 _a).'Tl)t SA65-l1 (J)t1 ,(= I, T)=r-i AI ))[1_{/I('J)I.'J'l!-n-T-:7.!.;-I;LL'.;;:n=-)l]} ･ . . 6 ･ . . 7 ･ . . 8 ･ . . 9 ･ . .10 ･ . .12 ･ . .14 ･ . .14 ･ . .14

5-Hydroperoxyeicosatetraenoic _acid

(5-HPETE)

#hk7 9 A

_{&JE3k%j=1)vo)}

n-:-.'!{1l

Arachidonic _acid (&) idi#7 9

_{xjkP53k%j='/VO)1fA}

5-HPETE =d;hk7 9

_{xjip53k8=xlJ+8}

SA6541 _n/ijA ju.:.-,(i-iL:i--,-,r)

_{I/Jt{l.:;-.,.}

:)i(Ii-J-i)-_,:3SA65i1 (I)_J'<:J)rP;

:Ti-S

_[llti

i,jf.,T;

_i:-.'')-I;,:-i

･'Tf

9

[ilti

-;;I:.I,:;;[Ti7

:Tl-_{2.;.A: .jl-r)lL}r=

-( = I, I)=r-i A1 ))H-/J(:771[r'l.'J!-n-T:;.i.;-I:Ill.i.::tTITi-SAG.=)-ll (J)jlililT:-_J-1lJ.'IilHl.:rfl :l'1)I1 _l'1fl-I1-)I--I)luL())_I;I_rplt.)

･'lTi-2

[iTJ-

SAG.,I)Lil(}l:i;.i-i:[f!.n-:Jf]-/T:I7 -/ I I)i.

+I/

･7'J-

:31^!TJ-SA654l -:1':_F_IIj'<LI)-.f$2,}) n'n_iiI!.II1

it.lTTl).i:.I-fl'::f',;

･':(i-1

filti

..-)-HPETE.:,-Tiit:i-.-i/-/11i-JirJI'::-TLl--,=t]L(I)

n=-r'iiLi

･iTil5 Itn-;-)-IIPriTFL.;.-[i-YET-(r>I/,1ui,):I':._:i-:)-I)L(t=i-J.1-}3SA6!I)-l1 ,J),1;JJ

+I.1l-･ . .16 ･ . .18 ･ . .20 ･ . .23 ･ . .25 ･ . .26 ･ . .27 ･ . .27 ･ . .28 ･ . .29 ･ . .30

･'l.Til6

[3TJ-

Jl-i.Jf))rE^':fT-)--7.( =} =t(-.i--Jt.{J:i:.i-[iYET-,-i/ _{-/JLRJT:._:t:-:)ltt}JL !t=i-J--.)-73 SA6{)41 a )_,1:)J

rll-･ .

.32

(4)

･'Ti-7 Ibm- #..I,:-Lli._L-.L-'J.I:.I;-i

:]1-_LSI;rllit!i,i-I:ii

:Ill;.'3.1;I.A:i.I..I,i-rT

i,I/1;

-!EJ-i-_I;ik_-A:.;

.i-iIIT-T: LI-)I-.I)lEI:'1ti:;;I:tliL))?1 I, ･ . .34 ･ . .34 ･ . .35 ･ . .37 ･ . .40 ･ . .41

(5)

ll1[i_/rl.

--li,-I.i:]

:-7 _lI)#unP&%

Ala

_(A):

_alanine 7 7=y Arg

_(R):arginine

7)i,qt y

Asn

_{(N): asparagine}

7X/<9 *'y Asp

(D):

aspartic acid 7R/<7 #'y%u Cys

(C):

cysteine -JXj=-1 y

Gln

_{(Q): glutamine}

_{9>1}t,9 -I}y

Glu

_(E):glutamic

_{acid 9h)/i,9 -I}_J&u Gly

_{(G): glycine}

9h'U _-Jy

His

(H):

histidine t=_Xf-}J

Ile

(I):

isoleucine j y tj

J1,J>

Leu

(L):

1eucine _t1-I_-JJ Lys

(A):

1ysine _{U ,}>}

Met

(M):

_methionine } 91*=y

Phe

(F):

_{phenylalanine} 7 I _I/i,7 7 _=J

Pro

(P):

_proline 7ot= U y

Ser

(S):

_serine k U y

Thr

(T):

threonine h i,*=J

Trp

(w):

tryptophane T,U 7o T,77 J Tyr

(Y):

tyrosine 91t= _-Jy

Va1

(V):

_valine /i U >

']1-'liE[lT._Jl;-':!I fI'[J+73 M.I:r

il-ju: Arachidonic _acid (7 9 * f<1'J%u)

ACE:

_Angiotensin

_converting _enzyme (7 y,}*f

_y-JJ2g]*##)

BLT1: Leukotriene B4 _reCePtOr 1 (p-/ = I, U _I>

_B4#-5%26Iq=)

BLT2: Leukotriene B4 _reCePtOr 2 (tj1 = f, U _I>

_B4#=5%%I4=)

ECP: Eosinophil _cationic _protein (W#ufR* 91*yJr&fE

_a(i)

FIJAP: 5-Lipoxygenase _activating _protein

(5-

) *o** _{,JF1+-iflfSJrfElkg}

_BfeBf)

HETE: Hydroxyeicosatetraenoic _acid (t= f<tT]*-JI-1 _=+j= I,7=>#u)

HHT: Hydroxyheptadecatrienoic _acid (t= T<'1tj *->/Lt7o9f>* T,U I

_>%u)

HPETE: Hydroperoxyeicosatetraenoic _acid (t= f<'tl

J<)i,**-J=j =+i f, 5iI _y%u)

IC50: 50% inhibitory _{concentration}

(50%PRSieLi)

IL: Interleukin

(JI

y9-t=-/ *y)

LT: Leukotriene

(t=JI

= T,U

I>)

LTA4H: LeukotrieneA4 hydrolase (u

-1= f, U Iy

A4hD7k/^iAS##)

LX: Lipoxin

₍

U

_o-JJ)

MBP:

_Major

basic _protein

(jlf2E2&Jr&fE

a

_(i)

MPO: Myeloperoxidase (_-II tj _A/I,**

-J9>'-i)

PG: Prostaglandin

(7of]

A F 9h)9 _J,}y) TNF-a: Tumor _necrosis factor-a

(Bf*#5EEq+a)

TRPV1: Transient receptor potential vanilloid 1

(/<=

t1

-I TI5%26I*9 -17o 1)

TX: Thromboxane ( fttj

_y1+y)

(6)

JS*

･'Ti-1

[iTJ-

Iui,):I;.:.tiT'l1)I:(i;1iirlTr.IYET(I)7t]:i;r'1h1_;.i1.l1)

1_T=.-/+LE-_iT_,)I-) 1ui5 /=L ,._L i, :)

7.:5:j1:;i-[)I5 JLQI _{-.)i;.I)i-)A_L1:i)..t)'J.Li:nli-J}

._Hii'_;lGLTt_,-iL. [klL.-;-iJ)I_{I_L}

Ult_{--..I;-i:i;ILJJLl:l[(=ql:j-:.[J-(})i.i!nli'-Lit.-:,)}}3. :JT-:_7-fL(II-tfJL[-l= 7/y _-Ii._,;.ii/Jt;i-.i:-_,:I)J1-}3,ILI'-:_.-I>._Li-)-[L= I/,

,//-Ii

:tL;iL(=J'l(d-;r)-;3 :Jl'I':._.-I+._L;J1;.i-,'_,:i.i-JJL-[= _-//7

-Ii. -...I+ii-tLtillfJt::i:-J)tifT'),.I:U)I;!;i:-,i:))L I-LtL_,4:(-:..-/JL_{L{.')'l-fE.if-TIJIilui}

1=:r'i._T:LI_L1

J),I r-:l.fiLri)).k'-LT+:i,I.TLiJrJl-:Ii;i-,I:.7->_.JjJ!_{I!)--:-:)ili.}

,l''.fit:_i-!i.1lB;1-)r-!ir..TT.. )JLT_I(i.:)-/It,.lu:.[t!liLil.i-:J(lJ:.)i;.;ielt_lJi

)J[T_ti[-)]T.[]iTi_'!i.-.)l{

_{)1-L7'1.:f.1-Jti'u3.k--Lit.I-tj':.(I:1'llJ-JrJl:.:)!l':-_:)}

i,I

, -j',:r= -H,lt:'-I.:1..'.;A-:1.:.-_Z1.I:;rT:.7' T,t=

-.I/lil.tJ.:.Ill

:j}ftLt.1T+

(I.;-.i)J!)it_);h}3._..)1-(EtiT'T1'lilui,I:!F:.E-i;)I-:'i;.I;:=T['rJl-fL;.)I;-/-:,(. jt(})ff;..-/+:I

_L}3I[iLT-)i'fJ--Jf))Li

I). -. :;1:E

i

--ti-a)!Li=nli.ill)i-!Li!B.i.

-LJ;-i::-tJJ,)I)i)1JIiU)

Jkff':..1-?/EJt'u)jJ&--l':i[[)i-()tl-.;iii;L -i,:)-I::.-/fL(j)ll.Y.'J]jT/- LJ:...j-.<.. _I _,})

ct5 ti-**0)fg;*f%k ritch-scratch cycleJ i PSAJ7flU'8 3). I nfESnf?tR-E=hlL18fS,R L,f=

]uiI]lk'(I.Li)_r,t'([;.i_:3:,_{'li-:a),).J[:-_.1)i-/)I;}₅

i:,(I.[u7-:[L:+}L..i /-I. 1I(I.-/A.'iI,-i)i-;FL1!.i=Yii'-LTtLi-,,i3I._L ))).i:-,.

lit/fT-,J)

QOL

a)fl_tI.-,i-A-[i_I:. ll)J/_a)I._i- i u. _{rII--:.-/+(hJ)lh'I;-ut}_{:ill,-.;".-.I,i:}I-_1-L;J3,V,-i,

iri-rli[/-LtiT'-r.1..',H_I1:1T:J)

-,)._LJ ill-/i/JJt 8). q-!i:I.1'!-Tt'::tifTJIJli1-i-lilLI'i(.IkJ.-I)-_}l_:i'-=Jf-)lllui4:I'::tifT:)..'.(-.]11i(I)r3H_-1t:h.)I;L!!

_E;h.-i,i

1

}3._

:lTil2 _{[3TJ-.1!-I(iiT'r.Jt:]i7ili:1LITi(})iE:ILL}} })I I/1ttlJ.u)

i-)L[t= j// _-Ii _:i;ti(/.)I;3i:.,1:)JL /lLIi_{1.n-fI'itiT'r.Hi1;li.TT--:a)}-it:il-Ll_,(_-))I _;/:i,i L -i- _I:i. (_{it)i-ili1![::iEfll:I}I/ r:I

j=-1 +-i (f, U _7o9-if') ,

,iA#t#P%f.rfEiE

a (eosinophil cationic protein

(ECP)

,

major

basic _protein (MBP)) , fEJr&%u#, +JI f,*j J (interleukin (IL) -1JL-2JL-8JL-13,

IL-31, tumornecrosisfactor-a (TNF-cL) iiif1), 1eukotriene (LT) B47bif1, t= _R9 -Iy

1).//Lu)Lr

-I-)} JL } ;)--tt₍=r-- _{I/-/.)I:riJJij-}_L _-LIL

"-i)i,i..'T.

_(i,

t= _-//,I

-:i _{HIJ1.'i-Ft[i;).jt;I.'rJi)I-}L -i/i 1

}3i-i,i.I-T.

L';-;I)7rf13:lF_;1''ttpiHj':fl'i;.)I;

(r'))L;.I.iILLT-_JI E-LT..;iL=-Jt L yL;;--I;/)IL1,_{(J)r1.ilhtL!'(,i.Ji.}:fJ(J+i _{1:I)i.7iE.(=tLt}-) -i, L,

}3i-i,i-_L'T.

_{(i_)-,]-}

_t=l-A-,(r,tt,I)f''lJ(i:1-i5[_fI_{ni.L'Iir,:LJJ.:/Jt;}_n-:JL-))_{L -IL} 1

73T!,i-.f-r.,i.Lltt7Jt;jlL'iJ,i:1kt

/h.73._.

(5),J)

j --)-1.l';,--- /I

-J) ') I-'.4-!i:(=7'--)lT- f,tt>[[r1-T'f(t..;]u1'y)(Figure 1) a _'I;R1-tlpi._{A)I''r:.Jij-(.I/)L}I

-i/

_b

< (J)LI-)I-.I)t)'Jt;fl--.i_-);h.-i/

-1<i=

_

1.ti.1t!lJui.1:(-.:T:Ii[t{).::I..1');,.'i...:I.1-.)-L 7' =>_{:T- f,tt}i.rj-fiJIL-[i]j.fi-JljJ.l.-u1--73

I//L --i -i /L + -l _,( I,Ill-ti-J-L-:JLf'J.'J-:_f^I-:)f):I))Ir;:LJIIT:-/Jt<i-.lr--lI!iJ.ik+}3I), I/,L =7 =7 JL +=7 1 f,ttJ(';J-,I/1_,1{

I),

,I--L; A= I;fl_I.i;I:jTii(IJ;B)-,I

r>-/1_:t=;r,I.1[ 71_-1)-I:/-/1 J1 i -/1_P.:.-Tii[i"J)ILRiML-))n-:).)I;JllJ.'fiu_iL-I

,iL;i I._LJ

･-,)/)Iit_,_:T)--/ I/, I/ > -/1 _{P (I i} _i:)Iui

-=t-Jf.)Jn-:.J).,Tilt:Hrj' I)=1--Tltt>["iT:i(t,:Llulr//))7)I;[',lJlj:)-}3 I

LL ).)I;;]-:IJ',iS

I-i:I;h.i.3_

Ij'_{=} lV- T1+}i _{tlI-;'f(j)(L.:Ll'Jf(.I}I-i.ii/r1 +l'(-_,iJf1---Lit

_L .,-,-I)7rt--tlt1,Ji--)--i;)I)I;tr._:LIit/._Ltf't

fr]kfA 5. _-J9 _{tj**-JF'+--tf'1ti#ghE=Fjg} L,

_{prostaglandin}

(PG) E2 Ej:7 9RE=jb'U'

Tla%#1fi:i35ik LifU1 6)75S, thromboxane

_{(TX) A25%3Ei*[q=}

(TP 7oP A 9 ) 1

f<15%3E>t*Iqs)

1f##

_(U-46619)

_{0)jkPq.a#175S} 10 _nmo1/site _{0)1itR%rC1S%#1fi:=d;hk+8} I t%, I

･})A:J.-.))(1-:).)I;TP Ill -//.I /

/( Tltt,17F(-1=1.I.:T;[.:i;r{LTtiljJ.l;-l).-_7JL)i I.iJ1 .-_Lli:):I. TP 7 T:I -/1I/ / 1 I,tt

Ji.fin.:'}Jih,rL/I/:Lit:-LILR-1tJl-.))n-:/J1;.tL).L1!)i:-,,ii )>LtL,_{I._L.}_/Jt;i:ti

;i.--LTt_,ii

-i/ _i,_}3 7)

)i)..LrJ'r)1rLTtT.[= i. _;]it

･t1---I)-i.I_H)h=n;:,t!.;-)I)I:Ji(ILr 3>+ / _-tr_{,( T1.r=it:iI.-LI}_{L -=L ,,:3}I._L _-I(3TP 7 1:I I//I / ₁ _{[,tt:/i.I;i['(tL-;,)I;}

L7-I)+ / _-Ill f,JJL):-i):

_{-;)(i.i!P,ti-Tt[[r.;I:F.:I}

1E1-LIL VLIL _1}3I._LJ_{;I)tL:,,TX^J)I;,)i-JI{} i/--7 + /

(7)

)7)HL-,_{I,lf'i_'r:..}jhiin'::_=i>t,-hI ..:i:I;E-i

-;./)J[[1;tT:f_(lil-I-A-);.n7n]

L -..J,iJ',E-i-,1--[t? =} 1 i _{r'lt](=irT--G:}Jr I/

+ / _{+r 1} _I,_(.=n-:)H_L

-i,,):-F.-/+,i-.-i-Filet.lil'[-,]j;.L -=L _{"I377rT'fEJL'Ii)I)I;-r5-fL}i:,AL7) 7)._.

5- U

_{o-JF'+-if11t=m4ht}

LT LTB4, _{-JXj=j=/i,t=j} = _I,_U _Iy _(IJTC4, _IJTD4,

LTE4) js'iUi> 5-hydroxyeicosatetraenoic _acid

_(HETE)

AS25Elth6n8 bS, 7 _9x _8=h,Ela

-1)-I/7, +/Ji. I/IP.-iTiicT,J)1LR-3<Jf))n-:/)I;LTBI Ji.'T7[,t(1-;!f[J[L-:il.{LT11iJiliIJ.-_tiL.i -/1-7-1 I JL [1 1 = 7,I)

:E i

,ti.'ti(1L-I.Ifl7t,iL:-Lil-i)1jJ[liIJcl I.jLtL, _{I._L} _)I),Lt-,. _LTB1

;.)I;.1:Tr-'..7>,J) 1eT'[:_:=nrJIj.Tr-73I._L /)I;/i''Iliflt

nTV'a 5). _LTB4 _n? _{9xJ-∼-0)&Pq&Sfj:,} _0.03

nmo1/site a tto-9 i LT##klitjB%

.Lllui_-=<JL'))(1-:,i..:.-[i-iE+-.)t73;Jt;.LT1) I_{-Lil-t=Li-(I)}h-.d7L-))EILti-.i'.,1!'_LT,)i/._LIL.,.-,) I (j) I _{,_∼-i.}LTBI _{),).;JI-:I.;;.';}

(=']j;JJtL1)1_IJJJfJfiU)

,A:):I'::1',/)''t-rT/Lit^>,,}3I LL_,iJ_,f{Ir'icT1-73.. LT_B 1_{[',1Jqf:!J7=-;Jr.i:-E-tJ}l{)..F]'

I)--? + / -1)-1 T,

I.:,F-8=#fRLTUl8 5). _{LTB40)5%%I4=8=Ej:} _LTB4

reCePtOr 1

(BLT1)

h'itFIJTB4 reCePtOr 2

(BLT2) _{LIIi.77(iL-).)I;i-,'}t)

, LTB1 (I klJ.1-j3I;:UFIT.IJ1:_.'tBLTI Ji.JT7(1{'J)Ii

I)

,.)I;I.:.iL

1

t<)._ _-rT-//1 /I

i _7J' _-'1t'LTB I

,:;TiicT,J)i-ui-=<J!-))(1-:I-iBLTI JIi:Ji;i(lil-.[,:i-;[:i',i-Lil.1iJIl;-TJ.-_LJi _{)3. BLrll1}

,I-Ii.7FLH;.,-iLJ dLl:A:

8=j=L=&RfR LTU I_{8 bS,}

-yj(EgB%1*WEEnPq,

qSi= transient receptor potential vanilloid 1 (TRPV1) _,Ii.Ji;['(,Fill-Ll[.'!-ljLIJLiTIl'}tT:F_(t=.i,icTl-llL -IiJJ ". _rllRPVI

Jr{(7(i=I:t7jLrli

-;I):)..h?ill.Ttlr.iT:

F_E-t.1:!F.-I-/,uJ)[Ll..;Jjl;I_{_trL:Lli[-LT.(i','}7LI._L /)I;LHi;!it3;iL-i/:i.JtB). LTBt a )n-:)f].L.':-J7)I;,LjL1:: i-lu TRPV1 :r,I.LUll

BLTI _J1.Jt';7(i=1c1:i-l[:t7,.LL'ri_{-1'):i.iV{i,1T}TII'j':f_-Lit.i-,'_,I-)_I.i _).)I;

,j'..[l7,ipJn Li:)-)T1 -/,,//i -/, P 7>Lt.tft(/)1ui.tJT:.iT;

-1eT1/'J)'[t'-i'Et-_-..I;:I:_{-;):!rL=VL.TILTS:lF_J}A)r[J:I)::(^J-:Ill!t=))Ir{LJi-)i!T =j+ /i)-J{ I,!=n-:)HL -i, _IJTBl

,i-1!t1.i

/I:.L IL-)i,/Jt;-;)(!ri!yLli:TLrT3:7!_,iJ.1.;uil/',L'(L rLlui-3<JL-))n-:/)1;,;.-TilEl iL_}L

LL !rJ-{Li:,,ii73.

phospho[ipids

phosphoripase

arachidonic acid

15-HPETE 12-HPETE 5-HPETE

, ,

;

i

15-HETE 12-HETE 5-HETE LTA4

A

LTB4 LTC4

i

LTD4

i

LTE4

l

1ooxygenase PGG2

i

PGH2 HHT+ma[ondiardehyde PGF2u PGD2 PGE2 PGl2

i

6-keto-PGFla TXA2

i

TXB2

HPETE; hydroxyperoxyeicosatetraenoic _acid HETE; hydroxyeicosatetraenoic _acid HHT; hydroxyheptadecatrienoic _acid

Figure 1. 77*

f;th>@*R>-

TI

(8)

･';i-.'3

[[rl'r= I(=l I, T)Ill i AI I)[[-/I('J)I.)!I!-t5iJ.:,J..;-,i-:/

-I/I/ T,

_L L /'=1-ui:)!['::ifr:).i(:]1'ti(/)il-(fJtEIJIi

rlJTJ-[ilti

-LTt7T{L /-IJ=') :I. LTB II-i;)[-:Ji;,;(I.]d7/J)i_L1)iJrJJJrlli,j)_Jk.1!L'Z1[/)'[.-T-LTtit-,)t). 1ui:)!I-:--tiT'1");,6:if3Hf-J7t(/)

--)a)1ri-tli[tLt:)- LiI+v T,-LTtii'}L.iJ_iL-I,-fL_tL;iL73

_. LTBl I:t7' I/

l1--I,tt>fl:1--:iJ[L-,-;[]](=JJtLT}35- T)_/1i

:1I--･>I/i1---L't;ti'ill<.ll..Ii-Lit.=L

,17lT-.if,1i{LTt;I(,,7) LTA I_).)I;LTB 1E,1'kt;Ii::ji:.-I)-i_L..i)i:'I.TA I))(J_{/I('}JY.-].'l!r=i+T:}

jfejS(LTA4 hydrolase, LTA4H) 8=i -3 ThD7kLDiA$5nTfEBk+8 (Figure 2) 10). I 0) LTB4

I_{:i,LLt)=T:4)/i-Jt--Lit.S,}}3._L

_i. i,(.I:+)i:I.'l::JrJ[.-.

} _-7-tt.(=r-- //_-,}) _--1

LL L -i/iXL)T--I;1:tt,,.-['_Il1'[TJ:1((/{[--IrJJ(),J)

jii-;E)kJ,i:J3i _-i)lt:i:,E

_(A;-).ill:

_{;iJ.i[ii!'I_}L I I. l=). _I

,hi:,(J)-;;I:[IIJJL1(})l1'r1_:LIL:.I:f)iLl{$l[lJJhJJ )[b::-i-';.,ir,I)Ll.;it---I)I}3

1n, _IL-_{a)a.tflTl-Jl'fLJ/lti;.I.i;.;l&IJ[:j)I;-;L:lil;}_L _{-i,.i1:.)J.r!:i.)I;)r-!hk}

lL-AL}3)+Lt

LLtt/)i:);i,I)jill((li'1:7')I;lL

It]l:Ll-.)-i-J _LTD _I

E:i/1]'-[lTT,.1:.ilLiJili.-i, I/ r= I/ -,.

-t>tt. I)--?-i-/+rl f,. T.)i ,,):A:,JJu!t;l.Ill.J'-;i'J")JLii>Lt._L1.rEI..

a);I':[fJJJh

-Lil;'I'iili5 _,iL71 1 I)._ _I';J[iT]-_I).i)-i,+!i-,.H-a)_{r1;J[lTJ-i(l'i[}_II _{1.n/)i.1iliJrJHJJri}_:3i!i)..

rT-,J)#..[.:JJti-l;.lilJ..:ilI"H-Lit(-i

LTB Iit,T3)_'t-/Jt;(ii:(:I-_(I I

-_

LTIJ-L -LTIJ-LIJtL). _{.;.:2:)i;I--:,J})

[r{r.:.'i:)fl.i

'!it;;-l;-L1+i'I:_q',/)i._q-.fJ[i,i-_{,J-.-1-ill.lit)tli:,}}) _L'llB_I_;)I;.i-.L!.

LIJ)i:,,iL-i/L 1}3 17)_ I (J.)t L:)!= LTBt Etr1'Ul/t![1-T) r>--.,+. .;I:2:)...::;l:-._{.J]ill-L:-/IJI!}JJJFbl-:]ftLt._i:J):)fl)iHIJIi:)ii

!i]..a_{),)[JJ):IfEi)r-I;)'h'=_,rr:llBtLt}f'{L'.I..:lJ

,i-+r.1l-/IL

-i, _i1_{}'J._i- -,tr,.}_fLl1,

,iL}3_

.Ai'=.]Li-:TLi.LTB I(J)J!-)))ul-(IJ-I.nL.1iH._L

J)IilJ)1LiflJIi<iJ_{,]-;I[:,ip1-}l}[]-)r-JIIi3_{i,I;:-ri;i.-.}Lt;It-=L _1}3 1L<'

. L 1'-I;.)I;-;i/..,-)-T);J;lV-t>i/1-)--t't3:fli(1J/TU)i

1

1tl;ii/)1,iJJ1-rJ.'[il)L 1 LTBt ,}));-t'i'I:_,1-)1iJ.'l;-u+}3.

l[,_L.( _{I-i LTBl}

,I"1.JiF-[i;.A)-;ill..[,:.'Ti,I:TLJ,i..:-.)-}3I

._LE-tlui1:L(I_7-,1L-L')lt:}ji.J:li,i.,/I

- I/:

tv T,.i-L!-I (J-L:!!tLt:);.ir/J._L/jLli)i }3._L rL-y,-{L_i:,;h)i.

5-U

_{o,JF1+-y'lBSg8nPRS#t}

LTEj:, 5- ) *o*-JF'+-V'FRf#, 5-1ipoxygenase activating protein (FLjW)

FRS#o)W#ji=bS11bnTh'

9 1 5-U

o->F1--tf'FRf#ASp#

.lJ.tifr')).:(,iL:_L L

-i/ _{I-_rh-:_t-1;h-i-i},}') m). _-E1=. BLTI _{Jt1.JiF'(i(;A);'L':.I.:}(_T<iJI;J1:.i..:-.)-)3I._L _{'iJH )'1(J(I}

=t_(I)L1.I(7(1=1.i.:f;[:]1ii-a)[HJiL7)I;.;iLLt!)tt,)(:i,I,Ii). );I(f_.rLl.U)fL:(M'y)(.Ir7)TJL VLI_{[:,-i"iJ,i:.;I,i.I.i:.j1/Jt;}_I)i:)j'd

I-_t_,h/i, L_1}3)I)I;. I,iL_{:E -LTt!.I}I-_lil-I-lb.;1L/-Ii,.j).I-ttL.iHM._.

-)J-. i:FBI a)_I_li_{t'H)W:-f:U,Ii(J)]li-,1:lil:'i[;1t-rLT.(1-:lf]} +8 LTA4H

_kPR#

L, LTB4nFAfEkqpSu18 I i Ej:1 BLTl isiUt> BLT2 0)f55%%1SB=

(9)

Celt MembrarLe Phospho"pids Phospholipase A2

･=.I.∼.

∼

5-LIpDXygenaSe (Oxidation Step).FLJW OOH

5-Lipo3<ygenaSe (Dehydration Step)

MotabolisrTl tO LoukoLneno a

L.TA. Hydrolasc

_..I.;I..

LTA.

i

l'!

LTD.

FLAP; 5-1lpoxy9erlaSt) aClivati rl9Protein

COON _Metabolism to PeptidyJ Leukotrienes

tT y-Glutamyl tlanSPePlidase Dipcptidase LTCJ SyTlthE)Ease cys LTE1 Figure 2. t]J1 = r. U

=y#a)4i%&&P8

Poptidy[ LeukotnBneS . I :i J.I .I: I . _I . . .I.: .: : .: :.

7 _-I/ & 610 &&75, E'Bk6 LTA4H (EC 33.2.6) E3:1_{--?a)fEgDIJF,+%f5-?/JJt1-i%} 69KDa

I :: . I . .r.. . _/ .: '-..I::. I I I. t1 . .I . . . . el .

%#&j?,7DT= f7-if i _{r6]#, di5;,t*%5GJ3to)%31-7*S} _HEXXH(X18)E i 3%g5ht @hg.

##o)MA bkM1 7 ₇

-I LI_-E=5}'i5531a 22)o3SWILTA4H 33B18fE,r4:qjJL.a)fE5;iEiHis295. His299h'iUtl Glu3i8 C=J= _{J3Tgali7+_53tTU} l8 (Figure 3)o

I I I:r.1.IL . I ,I. . /I I.I .. . . I _: I: . . .I :. ' I.

hTL ,_a 26)o _A 3

_{fBTCh3zB/{7tj51;,}

*##Ej.i<J+itf{j:=7Jio* -> T<1#jLnBAB#7?,A a LTA4 %rjl#

: / . ..i. / : _{L:Ill,..I..:::.} . .. . I . ..I. . I:L1'.:I I:..:[l. I . . . I i.:.. I .:I : - I I .I Ill I:r.I. /.. I I I .I. I. . : .. ll .I I. I : ".'. : I L:r.I. I I:... ../ .

a;#WhT= C12 1ji:7:jta=a i U, _{5.i(blh##8=EiA} _{6tL7>Llq*1&T.baa}

I. I. .. I I . .I.:. .. I. {.: . : . / I ll : I I I.

LTA4H Fj:,{7o5F r'1#%ktfDyF+87 i /J<7p31F>l--tz11fA]&#+8 I i b#%5hTU,8o

(10)

>c=ilTfE#1kghao =0)7 i )<7D5F.9+--eli;Jmi. 7JL,*)I/V.}A7p31 TI. f, u I-{7pjl T:-tj:i:.*Jt{0)a;#B=1fjiji-8 27). a:f=. 4%Ce4I[* (7-EfjI)liB4#L=F3aL#+aJ<7p31

I . .. ::I . . I. . /.I.: _.: I. : .J I..I I:[1.[L .:..:l!

5%fR LTV,a I i _{tBfr&fF3FikZ: 0)rWL*Lr*L=-?b,T*f5hTbl} V). _J<R 931>7)I; DNA%RE

k3fT"S)JL-C#PB6JifB*kqn*lJi-a I i fp E'i 0)7 i ) --{7D5F 91=iZ'1fjq i 0)rWL*.LtE79';,j%q&5 : . I. I. I, I. :.I:[1.Ll I . I /. : : I. !l : I I. I 7 ₇ u _{/5%#I*75;tJk#uB'aa)4?23<7gFklt=rfrtiJao} I > > 7 7 I)_{JIB:J=jk*P]B5JIO)te3#kq"$lJ} L

TU,a7)St LTA4H _{nJ<7uj191=-Plfjf]E=J=} D 4#FqT->17 7 Ll>0)Jj)h$7)S#= 8 tWB5TEi

t9#o)jirE1(=rW), 5 i jg=,-irj^8o =0)i 5 (I LTA4H _0)1#hTm#7Et %D7 i / _J<7p91

F=if'1fjl] i _{a)T#JiiE,r/+-_7);-yJk}LT)tit L'_a 25)o

I u)

cE I)C=. LTA4H E3. LTA4 3t LTBJ 8='Rf%+8 2S##5%,hta)#L u*c*-> flhDA(5}#SIf

: :tl . ..: :..I . I.::i.I.. i.J/.. .:l' . I .. ..I ..i .... .. .. I I:..I

I PEIV1)TL'SL^_ql>^n'L'RTK) ll.IIL Fu'SVnrTRFrn,T( iT^Ju_TVOSOF_I)N].A_qLVl.DTKL)LJ W

6 I LET(VVIT<(QEVT(Y^LLIF_RO_n'K(;n'hIE-lil_n^1 mT<t)f111li (SffTSPKSS^LQWLTT LID

1 2 I _{EOTSGKE!lPYLPSO(LQ^H} _{((lR^)LF1(1U[)TrtNKl_TYT^r} _VSVPKE _{LV^Z..MS^(RCK;E=l1'u} IMl

I XI PEDPSRt(1YKFZOKVPIP(lY1_ULLVVL;^l.F SROl( iPRT1_VWSL-KEQV1:K_<^YEFSETF-SM =4D

241 1_KL^EDLGGPYVWGUYljLL1'LPPSFPYG( iMFh'T'LLLTFYITTLL^u)KSLST<V)^l lEISI ES lm1

IL)[ Vr-rGNLYl7<KTWDZIFWutELr)lTVYllRHI(-I.1RLFt&EKF7L[!f7<^ulLNrliE- LU7<SVKTFGE.T)J INI

16l PFrKLVVDLTD)I)PDV^YSSVPYEKGF^LLFYLE.OLL(-i(;L]EHFL( ;FL,K^YVF_KFSY KSrT 13(I

All _{ll)DWKt)FLYSYFWKVDVLNQVDWT<^1nYSP(31_PPZKPNYr)llTl_1Tn(1LILSQRWrr^KJ.} 4H

lkl DDLh'SFN^TDLKDLSSEZQU<EFL^QTLQR^PLPLGl r[KFNQEVYhpF7<^rN7<SERFRW1-A 54u

541 LLIOSKWtD^1PL^l_KM^TEQGRVKFTRPIJKDJJ1^FDKS) rtX?JWRn-QEnK^SMl [PVT^ fun

Lru[ MLVGKDLKVD 6]()

Figure 3. t= t.LTA4H _Q)4#3528)i 7 _-I/#-&RF)

# 5 EB I'iio*.) r.'tJln*J<5}*1fBji: %u)} *=Xj> 21) I .. / . . .I. _./ I:lll.. . . . -:: !l. Ll1:I..I. .I/. . . .I..I I

3E*%0)6apEu;i5E7:,b8o %=-e, LTA4H Ei LTA4 a) C12 15tO)A(&&75; RISi: {j:9. i:f=

A6-.)7-A8- r. 5i >x-A10- ft 5r_>XI4;a)jf12 I, i)_=>kWyjiE+73 i: 5 C=hn*LjJLAq&Rd:&Ei-8o

=u)Lift;Fi##t=1=-97L3:} *=XLT.1 =*D*.>#v7) (C5, C6 J&) 75,E'#^f= C12

･ ....: . I.. I./:. _/..I: .. I I . /. ll ll:..: ...ll Xl. ..

Two)#3%7),6 LTA4H a)=*o*.> flhnA(5}*$1fjf]0)} *=X+LEj:i:^fa)i: 5 i=jfk 6^TUl

(11)

97JiT->-etb aBEBB# C7 15tj),6 C20 15ttiiFaldb7);-3 f=gE!rA-e.##0)f#SFj4<< i=+1 Ef 9 i Fj.i: io

-jft Cl llLL*/i,j{*-> I/- T. (-COOT EV7)EijEq;iqiI#+a Arg5631Ru&g7=,} r:last

&t&--( a/#%kjT3Bi-f8o a E)i=*h&R#rSLiEa)4E%Tjlj3iU( Glu271 A;1='#'ji'tj:7LJ/i,Idio*

L> f,'lETS15t8=4#ifE+8oi L T GigRtjt;iU( Glu271 1Au&A* JL,#'>#ii*/jJt+&/i)*i-8 I i E=

cE-?TfEJr%1t L, =7Jf*,/ TIh';FniSi-ao =nW5!. SNl jilt:bShE= 8f=bE=*/L,7Ji.*j1*

･l I. I. . I I :.:I / I I

L. ./(I.:.. /.:I. _.:. /:I:

. lt:.

:ll!::

lab;iFE6,r* sp2i:I#1f=i i. =c/) _{iE'Gfn7Eij1=-l].I-:-I,-}L+I rt U _I>gJC6 1iu)6 C12 _{I5u TC)E}

Ja7flE.L TU t8oAsp:r75 liC12(.1I:,-kLf&J4(FJ%+73 A('J}-7-%ffTWE.+8 i it(=tLTB4 n

_12-(R)

7k#B;a?1{/:(I-.T';;A.J=U< 'l/_14=#!Fyirr!)i,L11.I))[JJ5il/L{% ;riulnL+ 7D o i: f=. !iEJfff7:.A 8 LTA4 7)S#;,=#a)I-#

(R< i=h^)'HllLj=)r.S,-C.FBI/I:ll-:mi-73I i Eil LTB4 a) _{A6-.)X[4(JFflJ&E=iT4;u'=1&u} _,TL J3 _i

j%

i _L-)h7JL. _T1-tLti)t,I LTA4 0) C6 lab C7 _{15ia)PL'qi=EiE][IIBig4fiU)1;b730)-T..}hd}'dbLf=JrS-T.

+t1/;_ll;Ll-]+7DI i L=i: ∼##-EiL*##BE'=BLIT=0)B2#-Jj2#ht%& _I)x# V)_a)#EJ;ilkL*.

L .. I . I I I . . : ..I .. :I . / I.I I I ...-. - I . I.:: . I b8 (Figured)o G[u27

UJo

0

Asp37uO

__ o HvH I..,

r.tv=:I:tT--I

LLTA. LTA. Hydrorase

H2">H

}2"'

LyArg56,

H3"hL,s565

Elb=.i?--I(-

I. +. LTD.

Figure 4. I*f*->

Ti.hp*5Wffj7]Q)}

iT=XJ>

# 65F) 7 i /J<7o3191=iflfjqt%o)} j7=XL 21)

-jft LTA4 H 0)7 -I/ J<7D319t=elfj]jE=h'i187 i TI*%0)t^Wrtj.I +-f5;-1'.>>

I I.: _lL . . . . .. I . . .. _: . .I... ... . . . kE'hTU18. -:. I '... _: I/. : .. . I. .: I I 1; . I . . I

(12)

1fjf]L<i3 D I Glu271 7)I;LTA4H c1)A-/i- 1-'L)JE*-r(=f3tC173-=*1/JtI/319L-plfBjB= i: 1T

&*+f15iJ%TTkBu=+. if=. LTA4 i &f[T,r#.a).fI;LI L i) A-/31 r.1,FiAJfo)LB.f>t2Effo) C j=9#

1R1]0)A/i,#1*.> I/- rli Arg563/ Lys5<S.5i f[lii.ll-:)i]+bo a rjB=. I/ i / _{p<7o31g-iZlfjl]}

i= Ei Glu296 i3

ctUi>TyT383 /);.JZ.*-<tb73D +tL*jt], &FIt^WTg.rSIilu)7 -I h'l* /i/=Ji'=/L,2&o)

&#L=iEigDt75=6EliE L I

ZJU)

7nL/7T;i/i,ASC=jik[L T A(I("ijla)#%fu;1&ig a ^8 i: 5 8= Glu296

j5SA(/j7tjlk/I)*173o a L-)(I Tyr;'HuS7ot] L > Lt1+- i LT2&#tf9]WrEM_(Ea)7 -I/ A--y

t] Th_>&ii.i. _J<7''+ _{Lll#.Ji-n.U)L)JllMj)SX.=Tf8(Figure 5)o}

@H

-,0$2"H,tq

Glu271jJ..

_/

H2NTKHEyH#oH

1LH

LTA. Hydrol8Se H"'

_LyArg%,

wTRoH.

_H2NEFHTRoH

Figure 5. 7 i

_{/J-{1fF--@fFj7]Q)}}

*=XL ::I. .. ..Ill-I . .I

-:I I:r:1..Ll I I::..

.;. . I. :I.I;.: : .I I... . " I:.. :∼ ./.:LLL....t! I :. . I.ll. I : . . I... I _I: I. . 2 _{-?a)##LiLSE=j3'U,{A{ialb3tB94&SuktP_1TU'8,Et75S==-9} T>b7p (Figure 4 Bi tF5)o / . ... . I : .I..: I :I , I I ll:

iES#f5YLTU ,a LTA4H PRS#lij=C= LTA4 7+Tj 9t't a;ffLS*ai#EU+tj?Lt * L,

-j1 1 >77+p 9.i3J=UU JJ<7p31 rI#+ L,-f _-I y917+l= 91b4-jL=Jh5gh8 29)a

i a) 1LtailF[Jk i:1f C=ii%-To

IJTA4 7+ t3 9< i:L I Fj:ILTA4 _{a)iFf5f&&#-fb} T, 1)I _{>5BJ<7]ii,,<>iZ1>jP31*7} I >$

a)#*fSE=B& L1 a bC=**#0)HBryryRrJ&%7 U -/i,7/L,*/I,*k'2C&LT=1k%qbTtb

(13)

plc.(.I(r'i). 5 _mt,u/kg

_{,I).;T:F_}

[1J'<LI)---LTtJI'TL.II(}) LTB 1,J_{));'t'=-:'[i;i-I:IL',-;;-.)-i-J}:").

omoH

₀

Figure 6. RP64966

_a)#3E5E

),T{.tJ-iljiH::I,:I);-1'iErJ1-)-t=t _I/

LL: L rL,.'i/,-I -I/ ;+/ -T-I/-t'l,'[LJ.i-,:.:;;.i-,.));i;xJ,I+ > (Figure 7H=

I;lL7i,:n{JIJ;Jt;,i-,'_,:3I._i-_)J)I;;i-ri;i;.5;iL-i/JJTI). t= I, LrllAIH _{(Jut,17 :T-.i} TI))IJ7J('J)I.'J,'rJn-:)TH''Ti-;-)

[!Tji>.h-3I.I:i):I,i-J-+}LIC:-,''flo:I:i_-i _LIM. T _i / _;h</I-f- _{I/-i;1l-:TT] (:Ti-}_G

IJt3lt)--I,/1;.H.(J'.)

(Ii-J-i)-}31C.-,"fL]ll:E-t

0.3 _pM _7?hb8 31).

Figure 7. _J<R5fy

_{(bestatin) a)#3SiC}

･1--L,-1- ,(> I/1'+I:I //._L L p(,E:i,lILi3Tr.I;T:(J-} J} L7 -/ r-i-7-T--L't-Lit.i-,';3[[T.Hi=L> I,tt/-/+

//-tj EC3.-1.2L,1.ll _{,))I;llJ.;-.-::i...ivLTt.i;')3}Fr I/ _T,_{JL 7} -I. i _{/JtJ[-:L'J'-(rE-.'T['l(JI;[I:.i.,:i-,J-;}L. r1/I. i:-(J).i-Ti

:.I;t-.(.F)'Jt;Jl;I,I;.';(t=.J'j;/(

LTAIH _{'i-[,'LIJ.i.-:1-}3}I._LJ _;)I;.Hi;I.--_cjtiL-i- i,_{}') :W..} _{7' i}

i/I,1-:,--i.i. i

'ltlf,.-'u!i.1:.._,1,.i:-(ACE)

FRS#rC>b8*7o

f,7o) /I,8=b LTA4H

_FR$1fjqbSb

9 , t= i, LTA4I1 0)I*o*-/

T,1J)[1-/I(5:JY.i,'r!.n7Fll(IITJ-i-,Ji:TfI.-)

[!1t)-i,:<lT:'!H.:I

'(-J.T7-,-3IC,.-."fIL-l:[-ill _{LLM /)-I -:}/ :;/ + I/--L'll-:)fJ(

llTJ-3zB#

6

_BB*,q6)

_C=#1+a

_IC501#Ej:

0.07

pM t*R%5ilTVla. i17o f,7oU /I,Ej:LTA4H 8=

xLJlaR%bb7?4LPB9tiPRf#7?1b

8 33)

(Figure

8).

HOtN H

0

ke]atorphan

HS-∼N?.H

_captopriI

Figure 8. F9 I,)I,77yt*1

_{7,19)I,a)#3EiC}

/ i _/;/ + _TIJl-:=T-L,-:-7 _I >. I/i-I--h=7 I/ (Figure 9) -Lit.I;.,;3SC-57461A.I:i T)-l > t=t1->

I,L= _{T, LTAlfI}

i/)I.='-J-;:I--i..I,tt))[)/k 'J)1'.rL!.n-:)H'i.1C.-,HfLLfI:2..,-)_nM

_LJ[-:Ji;;;I.i-A;.( I:JLJ.;.-:L /,, ,i/

71.

-･J)#F_I[1'(L(i- (1O m!hT/k!r) .:.Ii:--' _-I1>_Jl'I)_'Ir,J)LrllB1_{,I));-l':A:/Ii,'i:L'tE}E7t_1=:=J-)iJ;lilJ+i,I):'l).- _;{>. vtt

917

_'}-/i,kE+8

JNJ-26993135 Ej:=*o*-J

_{T<'hD7kL^iWlfjB(A}

5

_BB*.qS,)

8=xlJ+8 IC50

(14)

15tj:12

nM, 7 -I/^7o919h'-if'1fjq

(# 6/cBB@,q6)

8=xlJ18

IC5015Ej:

6 _nM _7:>b8 35).

･ff=. 1-{[L:.'T1''/)E-I-(r>

,/,_{(I;r't(T }3}7' I/

=1Hltt> _{[l1-fiJ.-:,-Ti}hill-_{J'ri:.r.1)J.T!:<iJ-:i'::a}L]!'i.lj,_[rl.i:1

-30 _{1n!ir/ki;r.Lit}

)[].flT,_:(/(

().:[')'J:=JliJ.i..u-.1-}5._.

JNJ-26993135.':i 1'n _I-1'tL,0(.I;rJ1+7J -I)JL.i rL/i, _{1 -)-}/ 7 _-1-_{7'.:.-[i-1t:i)JB}

ILL--'i-} j=t (

:,---+)/-)i-.'_-ill(I;rJtL

Ill, l[,-.,

r>I/I _{7-'_T1'n[}_f'_,})_LTD

Ia)I;-l'---:!I:_,i-IC.-,"flo:o..'3_LLM -LTtJ-1jJ-''fil)L.

i-;[:fi.1Il!+i-7-t1(=L- I/--LTti-,'>3 lipoxin (LX) AI _{a)I;I(=i'(ii.=E-t'.;i.!'.i-I,i}L i_LIL,, _llrl1::i:!=

1'n I,1'trO :E---flt

JL -Lit.('t'}3 i):I1 :E-_-TJit_{i.:iTiiE11tiJJ[1li:]i-(}rlJ

,-'jE-:,--ttJLL;rJti

I-i, _{E_,}

/1.=fL:(Tliy30 _nlg/kLq Ej_{)}tT:f_}F]1',(LIj-ft

LTB4

_{FE4Ik#j28=qpSr]}

L, un4

_{FAfEi:,i#fSj:f=Ej:fBhD}

5if8 35). I)G-051 fj: LTA4H k

IC5015

47 _nM

_TC)PRS

_{L, In I(1'tTO%f'/I,8=3sVIT,} t= i,Aih*o) LTB4F%4Ik

IC5015

37 _nM

-LT.i)jJ_Jlil)+)3. i:M[-:. 1-IfL (T1/y)E-tJLtJ')JT)-fIFJ;ti-(})ifr:),.'.(:!'1.ri._LL -i/

-ll-II:.I-LTtlh;.T;),if.iL,i.[.LL.;i,'1',I-II1.l);)I;I-)ill))-hii{;IL

TV1836).

:Ij

o-moH

6:-{oaNOloH

SC-57461A JNJ-26993135

P.H

c.Momw.HC.

DG-051

Figure 9. / y<7pf

Ttt>#*t/-fJ(

yF7+p

Fhe)#3gie

･'+1'fLSIlfi 1<IT-)F')t,J)I [Jp1'J

I;1 L.i/Lit.ldiJi-i/

-1T/I L -:)(I. :.Jlf.1:lf_T!li:)i;!i-..._L.:.+.:i-i;:/LIT'U.;LL[;Jt;;i-,)i_-) i ;)-:I1,=r-- //_-a) _-/)-LT.,

･]a;.JJ_{,ill,klII::4:yJ''l--'E-Lit;i;'}i). /SJr.!1'J,,.-I'I_H'IL):.l:(/{[-[ITf:1().I.-Ti,)ln.:Ill_,j):(-,'i-J _LTBI J) _{/I:J:[(!_I^i:-)}_H

,1.i-llJ.i,-lJ1-}3I _LL._{E:Ei-;L:+Jl;,:1jlii;.Si-JJt.1;}_{Ultlui.1T:tiT'.")i.(,+LiiflrI.J}1[fU)_{JJ-L!!fL1--1;J,t._L}_7'Ltt)_fL7l')._L

rL--,-{Li:,;IL)3._.I ,ILi:

-Lit

-(,I.B-)7.)I(I) B11ll1 _,i.I.)F-H;A.[.:fJ[L-,]tL:(J)(?-.I.i).ii[;-,i.[.i-.j!1.)I;I)i:))-i-ir.

,ii,-I-ck/'=).)I;._"LTF:t=I-j:l'T,:-) -i, LufLtL

,._.

- )}-.

LTBI ()_{):IiLH)k.:iilf:h'lli,j)i-li-a'':il:'.W.-rLlll-:)I)-.)I)3}LTAIH _{,i-I;_U/.i.-:-.)-}l}I._L :I I:-' -i/ _JJllRt

(})i;'T:---:!L<iJJ)jJ.'[iu-.)-}3I._L. Ii. 1JllBI_{I:i.JiF-H(Lit.I-,')i}BLTl. BTJT2 a)i _I-I)tlJt_!=.i-.)-L -i,.i,

I;.ilf[].L'L!1--,L-';)..i-,'J)-)3I iL17Lt.(=i-a _)i-JJ[_{:}Jf.1[JI:-_}

Jl'JJ[.li./)I;1[:t1'.(.:-Lit -=<}3. jLL I -Litlt'-)T./I:i>L1-;[-:3i.1[LI-:_,iSiJJti U:1L;1i.JT=_tiT'r,)...:(,I.ii:

(n[tL'];'i:[I[h L LTAIH _<i-i)TjL)J:=Jl-H.-L;.u-.)-}3_{1AT():il(L.'M'J/J}_{LqA6.,-)-ll,iJ}LL[LI]1

L /-I_ LY^lfl _{till-1.1:_..I.,i:(1-I-_}

lliLfTm SA6541

_{niBjtJ+,#%fRiCk}

Figure 10 E='j%+. *Wf*n-elj: SA6541 a)37c3klflfB]

:r't

_L-Ltl.ui1-I'::JliJ;Tiun-:)TH.I

-) L 1-LI _r]j?..:--.)-_{L /I}

(15)

HS^HyRb

0 it

Figure 10. LTA4H fELrg5B15uliETCm SA6541 _a)fBE#E%*iC

(&&tj:4iigptL5+ kiF+)

(16)

#

1

_i

_##t=J{

= _{h U}

=yA4bD*3###FR%#

SA6541

a)LkJi3kn&ufFR

hJi-1

_I:Ti

TTlr-7EV)Elr']'J

I _{iL --I}-LitJ) t..)I.i.rU)j':A:lil-,-J} 71 7,7 T))i,,+iJ_,i

-I,1L (-Tq(/)i, L fL=ff'!i3ii-JJt:-f!iH=..-iJ3 pL/i'T[)ii

LTA4H

_PRf#

SA6541 k31H Lf=. SA6541

_{0)bt3k?ruElfjn=-jV}

IT7 9

_{Rjk#3kff')i,k}

RvlT1*51Lf=.

!'1Jf2

[ifi

F11 = I, I)=> Ai ))lJ/JL(//)I,'].'lI-n.E;),-.:-I,'AJi-::i;i-SA6541 a) (TIJk

Fi l]J!

i;tit .ILj! _,i: I(J-- :) :I :rT) f= t) , SAG 5 i 1

(s-(4-dimethylaminobenzy1)

-N-

[(2S)-3-mercapto-2-methylpropiony1]

-L-cysteine) k %BE

LT=.

%Bk}L,-

T,k Figure ll _8=,j%i-.

E2H-OysJyH

0 4 0 0

A

i i OH

sn;

snT-HSJ(

H

0 0 0

H2NtJ.H

2HCl

A

i ls OH 5

ilsnT-2

^=;_1

(SA654l)

msbovaN.+

_mS-koH

Figure ll. SA6541

_a)%BR&#

N/

F

SA6541 n%Bk 8=

_{Ej:i;Rfn##kjfh'f=.}

4-Dimethylaminobenzaldehyde (Nacalai

Tesque)

, N-t-butoxycarbonylcystine (Kokusan Chemical Works) , thiobenzoic acid

(Sigma-Aldrich

_Japan)

, methylacrylic acid (Sigma-Aldrich

Japan)

, p-nitropheno1

(Nacalai

Tesque).

;lI)-3 I.i;SA65il a) p

JI = 7, I)I i, A.1))[J/J(I))'j,'(!tr_1-i--,J.i:-r3Ilrl;[f)fJ

I)i:.tj:.).i-)}-.;if

(17)

''ii-LTA.IH E-t[-i-JL --I:-/ TJ].iJ-._Li)

_{itTi[':ilI'::l:-E'JL-r'ii!-}

_L

L!Ii:---,J,.i:-7'rL, _,((t=[n ,/I

I

LrllAIH I:JJ:.i.-:(^r-:)HE-t1)I

fO)jfi2i8=

ct 9 iP,TJELf=

37).

BEkL#i: LT 60 _mMHEPES Bg1*fi@ (pH

_7.8),

3mM _-}91*x I/j h-/I,, LTA4H h'

L Ult.i1;-([TTll(L:.'Tq'//J).),_t1}LUr3).+{rL7i1].iE;1'J/),i-_2({C -Lil 'JJ1-"I)_,(i

:T-=1-/i-.i :Ji. L /-I._1L.I):(I.50 _mM

-,I;-I;).[.I.i-,})/I(r]l-7:iJ(i:I)+ rL)i(i- FT}T;i-i,IJTAl (I);T- )/ /

-JL _{I;Jtii(''<-,i-))IT{L,}

l'fi-,1:Sn'J:I60 LLM (I)iJl"lJri:r-i L

1=. I(/))-i].i:iliL:T4lJ/)iJ26t-Lil 'JJ1[".j1 i _{lt--=t_J{-.i.}I_Ii L _{/'=fiL t]1-7'!.1JI:_7t,}TtI- _rJ) _{)L(i;itft(J,i}

(7*T,=f,U/I,:

I9)-/I,:Bf&h =150:50: 3,

_100pL)

khDk8=t8=iJ3Tji

):Lt:,i1.':I:.[T-_I:i-a-.Lit i:,:I. _{I'::;jrrJiti(.''/:(i;}// r1

-,, T, ytt=)

-/.I

-(HPLC) 'J)1=)T.,J)I),JA,:I:1rl.!t.i(i.tL -i, _PGB3

kADkf=. I

_{a)&FSffa%4hk-20oCTe)}

30 LAYpqlR6t 7j(v'T) 4oC7?A 5

JZDYpqL#JL,5iil#(10,000 x

g) kll-3f=. i.I#0)-E% (25pL) kHPLCTh/^tWLf= (*9L :TSKODS-80TS4.6

x75mm, ##fB

_{:0.05%EDTA%E7*}

i,= T,U/I,: }9/-}t,:7k:Bf&u =900:300 :

800 :1.8iR%f4fiTRpH5.6, i)iEL#: 1.OmL/ LAi, 1*EH# :UV 270nm). LTA4H FRglfR

Et'I=_I)qL /I LTB1 _{;L{'/ii-LIL.,_:-.)-TJ}I.i (I I:-) _-IilLuJ,ii'L 7'-I.

._EjJ-:1 lC,.-,"T[t'I:f:i,LlfLt(._L

l[_,

5d)/1+_L 1-I:3

iJ1"ttJ_tri:(:;r3t(+ }3Tll.'[rt:J7l{J.i:tltI;;:1-I/i:_,Ji:2 It7TfT.L,.

a''.'itJI--I;rril.).I..

/7)11:I(-:.I L-) -i, '[:;.:J/i:L 7-I_.

I-)i'-.[.1j.;iLl1-ll

i-SA6541 0)%}t,f.y I,_Eb* LTA4H

_FR$1fjq

SA65L11 i-Jt.i:UltLrllAIH r:n:.i;:rI-:)_f],11]-T7-,--JI iI/, I/+ i ,})_{L'llA lt{ (}_I.i-J--.)-_}3_{:tLii:g',/)it.Tit).li--fli:--jL:-I:I}_I.

fEb#h'it>.PRfWEE

(IC5015)

k Figure 12 8='j%1.

100 90 80 70 5 60 -J3 i 50 = Be 40 30 20 10 10 9 8 7 6 5 4 3 2 -log([](M) Compounds IC50 *(M) SA6541 2.7 X 10-7 Bestatin 5.5 X 10-5 * _The

IC50 Values were _calculated _{by linear regression} _analysis _{of dose-response} titration.

Figure 12. SA6541 h'j:tFJ<R591y

_(bestatin)

a)t=JI = Tt U

IyA4hp3}##8=

#+

8

_{#4b*&-##FRSgb&h'}

3:tMC50

L@

(18)

SA6541 Ej:LTA4H

kieJihk#Bg8=qpSfJ

L, %n IC501#Ej:J<X 9 91yo) 1/200 _Te)bJ3 f=.

A

4

_BB

(5-HPETE)

idi#7 9

_{RjkfE3kff'/I,0)1f}

r'iil

Jji:.I.L-.).i)J-i1-;

flit-_{TTlI..I;riYyJ}

(5-HPETIL

Biomo1) kRV 'f=.

I.Ju-)M!])1'y)_{:rJtL -i-)lt]T!.,rT'}

J,-I.:;-i-ll:i::ti:Ill:

6-73A#0)#Jr&ICR79Rk,

12

_{FSFRqO)PjP#+j}

9/i, (7:00 E=JE,*T) f, iiR23iloC

_fJtL -Lt;titf[l),_I_lit55 -i=10t!"(=nl--' rL1{!..hJ[-L i=. [i_!1TI-_P.).ii[.1l::I.._L/J(E-iL'[[IT:I-L]-{L7-I_ 1)i:.[1L;!f_lL-JL.i

i

=1t--'Ili:.:i-(J_)1-)J!.

--(!=fLt:-/Lli:I)JhfL 1=I

5-HPETEW

5-HPETE (0.1% _ethanol _{sa1ine9a#iS)i)} 25 _LtL k 30 F'-,}S+0)7j 9 tj.JU y-}kjB

i,-i, -,,

r;/-/Jr.)'Jh(.I_r.hJIJ.ii:_(H.L 1=. J!{I)i_ti1[il'H-_('iit;Hf!J-,l'"'HI _{TIBH-rJ).rl...:()}_{))lit-:[l.i-JILtLL;}_{:I J=}-) _--LI-jt:i_//LLt_-a-,

IJLL-L['r'r,I:,.I;l;(.rl:,i-6 mnl j/:///- I:, f,, l> +-L111--i-)JJ{--3<.iltl:=l.;i,0..,I)t)u/-T--7T:)--tttL.JL f, I)} +/LT >

-i:-I- r7)i(/ L-1 -: fI,.'80 nlM [)i tlTL'i.a'':i:-iJ.-iit(r'iO.7{) _tnL _(I-I=]-- rI:ill. _--1-=tl.i!ytt-t'ti[E-JT1:_il"I/ii.)fl)

b8VIEj: 1 _mLI9 /-/I,

(LTB4iRr]jE'+jn*E=#fELf=o

-Ii,-- rlJ i,L _-)-:'t--_t> I/-i;. (MPO) _{I;._l-1'l:.u)iti[J'Ji:}:LH)

-I)-i 7t ,L_,i-,1=:[-tJ+ /(-/1tt(i-:JJ[l-'tl(i:)L i2 W;tli-*-.I.:._Lf....-.I-JJ.I:').Ill,Ji:,i'Jr*i);]iL /-=fi1.15 I)JIB"rJ]i!JL.'J)

-I:lil'I:(12.000/fI) ,iJfJ---,-/I.J._tiL'iU)---[;--I:(.'3()LLIJ);i,96 I/I(-/i,

- f,

[f'. ])i _{fif!a'':J2iJ.-ill:_i:FIE.f{l[1",;}

/I(loo _{LLL JJt i:Ult}0.i22 M T)i _{rl:I-;1S:l''..)ifJ.-iti(r'.:-}7..I)_LIL _-Lit.I;fJIL. ;)(i.I:i-f,I).J +)L.:{> _tj.jt> _,'']lB5

#1kA(#k%&##2Sff

(Amersham

_itA)

35 _pL i 37oCT. 5 LhFpqffa%Lf=. 1.0 M 65BEu

I.i()LLL ;1JJ)HfL-i,)I.{J.t:,iJ(,I:)-.([-_L ;'T-7)-iI-/,L U'ilil;-i.FM nm (I;r)LE+}')[1)i)t)_ti:,1---I1 I/I:I !//(I/

-7t L

-TJ) _-.:/-<iJ)fJ L_{I-i,il-.uJ)i:}_{L /I._.}

tiff.ft!IiE-tn-tL-)[)i-i1-/Ill-)/Ih,i''lL;i'l.'ili(0.2826 cllltJ)(j)rr'lffL.i=

kjEu'T#jf

L,, OD/min/cm2 i _LT4i Lf=.

LTB Ia)iulJJ,ii'

+>7p/I,k*f-}+-1 R1 (i9gflb) L, 15

J{5}FpqL+JL,5}#

(12,000Xg)

kllJ?f=. _{_tf#i:}

i1.-L;k)I-:1'-.;.lLil;I,1'[-.iL 'j.k-'L:.I(1ly),iJ0.1

lT.II/;L i ["l-:'i.-)-I, I)rl/i,:1'J2iJ.-it,I(L'.(-(pH :3.0) :3111L:.I(i:F')JL'J'L!.L /-I...I;'i:i

(19)

i;Lt.1',-L /:I llllBl I-i1"(,LlrI;ll!1-7'f

(Hll

80t!;,i I/ /

-,i _{7J((i:i.Lt(-'i-LTti7i.'I'.}L ill"ttj'..'f[-.i.;J-_LLr.I.-]_,i{i]--JT'i=_. I J)

f&EHkiATlj:

LTB4 nB]qR-*Ej: 89%TC>b -3 f=. LTB4 Ej:##9a4i7 ,ykj *.y T,(Amersham

TtI-_.'itn

i.)I]L_{1--i,J,LIL.'J'..:}L /I. LTB1.'I...:[-tl-J--i-,i-)I:L,/'=1l-)'ra'':LL;;'[.'Lk(o.2826cmq (})[(lT-ft.i,i-)llL,-LI

ltITi

jEL, _pg/cm2n15t LTafLf=. I-fi--.I.i:I;^lh[ f-+I.ii.

5q

f.FEI,,

,I)iJllt{t).li:a)_5-HPETE _{IL.Lip:tfliJ,I.li(l'{L}

,iJ-i, r')_-/Jll_)'riI [[{[I,Jiiil'tIL -/Ii [LIJ-:rI.I,]fSia)/i(-I_II

).:.I:I(I.i.tflT.lJ(A/n)0

ffJht

_LT)

k Figure 13 i=,j%i-o 5-HPETE _{(10 LIE/Site) 0)B#8=iJ3TPB} 6b,ti#*#

it.-i.tll..TJ;)1;.i-.LJJ!)tt) ,iL/-I.. CV E U LLh = i a 0 > > U O3 0 1 2 = C) U) rO a) L U = 10 8 6 4 2 0

Saline _{0.1 ilg} _{1 pg} lO[Lg 5-HPETE

Figure 13. 5-HPETE

Bat

4

_{PSRB^i&o? 9R#A.F∼#q)*i2m}

)IJ:(=t ;I)-HPETIH5 LLgrSite) )t{LJl.;1--_l'H-fi2,))f[r.-_I: ,I)Ir!)-:l'"ll(t=J't(T ,-3/1J'-I_{I'l:J:tltri.tll'.[J}_{(MPO.ir;.rJli}LT L -i,₎ _i-_Figure 14 :I_{,J-,.+..I+.{L),)ti}lilrfJ-2AI TT!J-,h"']fii.:i-LTJLJ'.'II)i.J;it..i.I;l].I--I,j_)lL;-',I))I]).)I;.;-.i'.,1!'i:,Ai/==.

CV E O = Jf C) 0 > > O EC 0 1 2 C V U) CO C) U C 4 l2 10 8 6 4 0 0 4 8 12 16 20 24 Time(hr)

Figure 14. 5-HPETE

_{(5 pg/site) Bh1@o?}

_{9R#ALPg#EP*@m}

(20)

5-HPETE

(5

_pg/site)

jkPq.a#J1&o)#AtPq

LTB4#jio)tB-k

Figure 15 8=,it+.

#*

F)iLTRl ill"lJ.i:.-E:i5-IIPETE _{ti:tLj'llfi'il.ii---(').).(I}I._

rT['-L _tiiilrlJ-2 _[r!J-:ll"I)i$2:_=E-tfltLI _L

,i)L :I_)jiJ?1-I,

∼ E O D CI C .9 O3 L C Q) U = O U tl CD L J C a) U) O3 q) O C 00 80 60 40 20

1y

i 60 TIme (min) 120

Figure 15. 5-HPETE

(5 pg/site) B#@e)?

9R#A.Pg

LTB4*&

A

5

BB

bachidonic _acid (ju) =#3E7

_{9R&f53kfj=1/I,a)1fA}

I)i--.t.j:.,i!J}-_i1-:

I,Li-)I_1,iii4J,/)

s.diim

arachid.nic acid (AA, Sigma) kjn,f:.

I.i-IML'])1'y)

-1'tI:-(_)ll{!..1_l!).-i)ilo.[;i-.litf'f

:

6-7ig#o)#.EEICR 79Rk1 12

_{FSFRqnPj]P#+-1}

9/I, (7:00 E=JE,*T) f, iiR23iloC

_fJLI:Ulti.[lJ),_)tit:55-i: 10(I./,:=n''-/i,i!J.hJiL /I. L!+i))L;.'-!1{!..Jl':I.._L:7)(E-iL'I[h (.=1j-/i/-I_.I)i'-.-5-.]![-i;JL,ii

='(-JIJi:_L-iLU)i-]Jt-.-i-(.IfLt:-/I I)i:)j'diL /'=_.

AAW

ju (0.1% ethanol sa1ine&igai&) 25 pL k 30 91--}$l-n7j 9 t1

-JU y-}kBijB LT

-i, TJ7I/,1r

)'r.'=l!{J),IiiiJ'1J.L -/I. rf{fJILIIi]'1J-i>'i.;.'T[[!;:fI"'H=.;liMul.-ti_:,j)lit:l'yILt;.;i--:I t ,' _-i,i:I{/JE5

-Lrt I/LL[r.I.I.T:

.:;--r',I;I_;i-G nln1 -)/-//)- I./ I,,,i 1--LT.i-r]J_,J'){'it.

.i-'tl.:I.,i-()..5-/'.I,;i:1---rr--)-:,iJL I,))} -T-/L7' i.

-i:-=-9L7'tj _-I

_fill/80

_mM ) yg*#'tffiik o.75 _mL (_-I_{It=^/I,**,>F'-iflfE#iRrJEBn} b8

UIEj: 1 _mLI9/-/i,

_(LTB4iRr]Ejm

_*E=##Lf=.

-II _{tj</I,**-J9>1-91} (MPO) ffJr&0)i,BJ]E

･Ti-1.L:I,-:;Ti--1

[i7rLTtjLEJ

if= )/u{LT.i)i'-I;kL 7'=._

(21)

+y7o/I,k*f-i+jR1

(j9({1b)

L, 15

_{LAYpqL#JL,/^iB#}

(12,000Xg) kll-3f=.

_i.I#k

ilLiklT_I:I;JIEllii'[l-]:L 'jk-J.:.'!1'y),i-0.1 M //=tl > _r"I-7:i+T,I)rL/_{lla'':I-i-fJ.-iil'i}(pH _{3.0).'3 mL} _{(.=tiJtF.(.i:l!-}L /-I._I;'ii

ilTkkC18 SepPak A 9L

_{(Waters itA)}

_i=BS?#qL,

_1%5f&u%5

25%} 9 ) _{-/i,A(iiilRT>}

I;;ti1'tL /-I..PCTJh f:ir,I;,i1'l-:r'tI-:.ir'fT(J-(sot/'(,,/ I)/

-JL _{/I(i'..:ii(i//LTt[iF'.I}[I.L iirll.;It.'f['i.:,:Zr.I-Tl_,iJI(_{j---_'}_{/I._} _I _'J)

fbEHki2f7?1j:

PGE2 0)FqX*tj: 96%ThbJ3 7=. PGE2 Ej:#iFiA#7.yk-1 * ,I h

(Amersham

_itA)

kFBU'TjEr+a Lf=. PGE2

%Ej:fT6ikvlf=#*,iR#t#

(0.2826cm2)

0)iii

*kjBvlTIGjEL,

pg/cm2

n15t

LT#Lf=.

'ji'-.I.i:I;illr,:!li

AA@

f.r[I,,

(/)it.T'U!i:a)^^ L.TLtf.:tltYJ'.li(L'{-;:,i--: r>_-/Jr.J'H.I)

+l{r)'J;i--_]'fJ-L /I L1[T!J-:I-.'')(I+1

())/LJ'-L_{ITli.1:tI1,i.tllT.'l}_(MPO _ir;J[J[i

i

_LT)

k Figure 16 8=,j%+. AA _{(1 pg/site) 0)ii#+8=iJ3TPA} 6A,ifW*Z#iafW75S#hb E)nf=. ∼ E l4 U lt C AE 12 tt C3 0+ ₁₀ > = 8 O rE) 0 6 a_

!

4 q} S 2 C> _e o Saline _{1 pg} _{5 LIg} 25 _Llg AA

Figure 16. AA

_B&

4

PSFRl@o?

9R#A.Pg#*BRia.jR

;):(t=, Iu (2 LLLLr/Site)rr'(LJ,Jt;_i:_[1Jtf>'it,I)fLTL:JI

,I)H!J-.[l"'H.=J't([,-3/1J'.-[I)::I:I(I.i.I,ll.L-I(MPO.i.Ll-.l1:_._LL -i/)

･Ji-Figure 17 (I :l=.-1-)'{[Jl.iii1'H-_{LW!)-,fJ"'JrLT./i(-I}I'l:J(;/i.tll'.L1()))ll'.I))I];,I;LIL- I/ _{_L} ;+LIL). =tt,})i:,'itil.!.(,i,IL

(22)

∼ E 16 tt E -i 14 i

E;

12 ･1? 10 5i 8 0

%

6 E ₄ 4) U) 3 2 i O S 0 0 4 8 12 16 20 24 Time _(hT)

Figure 17. AA

_{(2 pg/site) B51@e)?}

_{9R#A.Pg#*#iaiW}

AA _{(2 pg/site)}

_{&Pqfi#J1&o)#A-Pq}

PGE2#Jio)tfLk Figure 18 8=,j%+.

#A-Pq

PGE2

ii"1tJliI-I-i_ju _{tiiil'1lii1.:Ihl-I('?.).!=}I-_[,l'.

L _{ti_iL:]'H-}2 _LT!;:I-"]Jfii.:t=.'ifltL, L _I-{JL _(=],i-'f=,

tT E U ∼ a CL C .9 EO L C a) O C O U CV U O E1 = A) U) O5 a) U C 000 800 600 400 200 0 0 60 T(me (mln)

Figure 18. ju

_{(2 pg/site) B5?@e)?9R#4TPg}

_PGE2i*&

A

6

_B5

5-HPETE =##7

9xjifE3kL=xL1+a

SA6541 n,ijR

I-)i'-.I.b'.i)}-_i)-;

frii-)IJ.i.'1iilJy)

5-Hydroperoxyeicosatetraenoic _acid (5-HPETE, Biomo1) , SA6541

(@j;A##iCi>

it)

B'iU:1ndomethacin (Sigma) kjqVlf=.

I.ii-TTIJIJ)1Jy)

I1,I.iUltf!I'1T'il-.I.lrU-[:i-:ttf1..

6-7 _{iq#o)kBJr*ICR} _79Rk, 12

_{FSPRqO)PAF5+/}

9/I,

(7:00

8=,E,iT) f, iiR23iloC

;!''1tU'tti!J!!_Jii:i-vI)j=10''"(=n1--'-i/1'iq-lJ_iL /I_. I.I.i)"I;-i.J.i!'JJIL:I.._L7J(I-iL'TIIH.=Lj-fL1-I._.I)i'-.Li.)'!t

(23)

=V-'[ri:.-i-(/)i-,rll:.-:.-(t=I('t:-) _-Lli'.))-'dL -/-I._.

..-rHPFJTIL:iJf,:iET-,-> -/J1- J'r:fi,I;I?_I--7:1JL

5-HPETE _{(200 pg/mL} 0.1% _ethanol _saline ABiBai#) 25 _pL k 30 F'--}S+0)7J1' 9 tj

･> I)i.>tl;'i:Tn,-I-I r>-/I_{I/LIL;=lI{I).Ji}_i:.[H-_{L 1-I_}/1j'-II

rli1:(]H:t.'i-())IturJ';i;)l]

:I. lil).lti_iil'H.ffit8 _H!J-:rI"']

-Lit;tMs[JltI.i:(.))Jif-:tq-'IL!1ii(iI:-) _-i,i:-[{Ill.=1i-. IILLtrTr,1:iI:(i(:,i-6 mm liI/I jr tt/ I,^ i +-LTtJ1-i-,i-){'_-=<.

,i-'t

rsTk 0.5%J1*+j:I-J)i, I,U } 91)I,7yf=9L7't= -I

T<''/80

mM U J&#'j*if& 0.75 mL

[[I(=TllTrLrn;L /'=._LTB1, PGE.1 J't._I;-G:LTC,1/DlnLl it.uJ,ii')I)(I,)I{tJ1]fi:_[H.ff130 '77i-L';t]lBW[-[J:Ii_:a))1l-I:

t"iJ]ti;;,i-(.I.L1 _-L7/[{/JLiill-Li-,'JiiLL7T7:.:;-.I;(1,i-6 mln }jI/Ii)-_I:, _T,,,i +vLTtn- I-)J-)i-5I

.i-lil::I.,Ji-1 mL

I.--I/ / _-JL _t[T:=[f;1')_[..TIT.':_L _/I-I.

SA654

_1*

1-J-.rI:1.i.-iT-I)7J=E _{:i.:'1':lF_}r]J'(LI)--H])tli-)lf[J(J).-/Jt<i-itT:F_TJJ'(LI)--_L

:.Ill.i1'y)1'{LJj-:(r>I/I_:=f_:i_SAG.511 1()_{;r3 E}

U( 50 _mgnig, b8VIEj: indomethacin 5 _mgnig

_k&R]2SLf=.

_&R]3;ii-

1 _FSFRqi&,

I,-)-HPITllE2.r-)LIL ;i-30

J]iT-,>tti..:I-,-I)-(1 I/r=t.i T)i _tJ,'i:mL,.LI-,-, _'J)

IJJ1-)'r(I)-{r),lit--_J'H-L /-I.

/LJ'-r_flT:1:_{(jifL-i:J)iVuJ''ii'(}))-!"i.'T.I-i.}_5-HPriTE )_r{r]iJ_{ii:_l'H-}2 _[T.lJ-:ll"l]f{2i.I_{rTJ-.J'ri:-:i!i-qJ,)}_{,1-i:lf_}IIlJ{Li-i-L 1= _

-II _{t=<}L,**-J9>1-9'}

_(MPO)

_{ffJr&0)iP,T]E}

:1L-

I _]:I.I::Ti-i

_[iTJ-

_{-Lit.lil,{Ip:})J-.i)-.--LltJ:li'-))J[u-L /I.

LTB4t PGE2 A- itF+ LTC4ID4/E4 nfRTJE

-tT> 7 _JL

,JiT;i-,:E-.i:I+/(I/,tt(i-:J'.tJi-(L:)L 1.5I)JIL"]J.;ifJLt:I)'l:I)iE(12.OOO,'gr) ,i.7J---'7'-I-

I._77i-;li-l:JL.kl_I-:l:I(1.i..l{1'IiL /iL-J[:.71'y),i-0.1 M '/=t1 i.[1-:i-+I,I)r>1t;i't.:L-i-fJ.-itft(l'i(I)H3.0) 3 111u=T..:i-I.j.'J!.L /'=-.I;I;i

77ik C18 SepPak *9L

(WatersitA)

8=jiSBgL,

_1%W#%E

25%} 9

_)-}t,**f#T.

#* Lf=. LTB4,PGE2 h'

cEU( LTC4ID4/E4 Ei

1%5f#%E

80%} 9 )

-)I,A(i#rC)iEH

L,

i*n%W

k4iJ3 f=. LTB4, PGE2 h' i U( LTC4ID4/E4 Ej:##A4i7 ,yizJ1' * ,y I,

(Amersham

its)

kjnlTjE'+i Lf=. LTB4, PGE2 h'

ctUi+ LTC4rD4/E4

%tj:fT6]kvlf=#At#R#

(o.2826cm2)

_{0)f5*kRU'T#jfLt}

_pg/cm2n1#t LT#Lf=.

･ti'1'j.i-I-(.'J.'f!1)T-i-J-.tl.l'.iJl.T:._L_:;1L:1'yJ.i1(L I)--_f1.l'-:(/₎

,t[:.::E-tI)uIlnet.t.j)/S.r._:i-LL1.:.n--.;r'i:ii,)I]LI-L1.'].'lI-n.L 7-I..

I

)il-.lL-.;i_;I:.[.:

-ill-50

.rI)-HPlmllL.;[i-Yet--,,r>-/Jr llh:+3f:)Ill::(I;r't:[ }3

/1J'-L_II_I:Jl-i._i.I.l!L!--I_:

I-klJ.-;r-,-3SA654l J )f1-:Ill,Ji-Figure 19

(24)

8=,it+. SA6541 50 _mgnig, 2 B]0)]2SEj:. 5-HPETE.a#J 8

_{F#Fpq1&o)W*fR.Ixfa.mqkRi+JB3?}

B9E=#%8=UpSu L7=. _-k, indomethacin fj: 5-HPETE =di#7 9

_{x#At3krLE8=h'E18ifP}

T.i(tlt.1tTl',LH=kJ.L -i._{I.;:.t!:i.",:J1,}_Ji_Ei _i3_tL7).-) /I

_. A ∼ E O te

=Eq

E) gi 3h ∼ > ∼ O F* 0 i = C a) GF) ∼ O h U C 8 6 4 2 0

Control SA6541 SA6541 hdomethacjn 10mglkg 50rng:'kg SrT:g..-kg

Figure 19. 5-HPETE i##?

_{9R#A.Pg#**iim8=#+8}

SA6541

_a)4fjq

**RO.01 vs _{lthe control} _group

5-HPETE idi#7 9 A

_#A-4i4iEE=b'

_Ela LTB4, PGE2 hl

cEU(1TC4/D4/E4 i3;JitfL8=xl++ 8 SA6541 _nlfRk Figure 20 8=,j%+. SA6541 50 _mgkg _0)]2SEj:1 _5-HPETE.Zi#1 30 JAi

fii"J) _LTBI

;rjtt U: PGJi:.i(J.)J-_

'Tlr.!1J;i'7Jj.ill.;;::Ip]t)::()-.L1-:.=11iJ.li-uL /I;,I/;.llrC _{I/D 1/Ji:}I(I) I-_'T(].:=t-t'r;!.-;

*4k2i8f 5 tfb,-3 f=. _-f, indomethacin Ej:5-HPETE =di&k7 9 A

_{#At3k3fE=is81}

8 PGE2

(J) J-_

'rl'-,1J;i'Jj.i-J.I:;::r'J'J:.I(J-iT:(_=i-liJ.'l.-uL /I/r;. LTD I;TJtJ: U( llLIC 1/l}1/JjI(I) I-_L)-l'.:=E-i-L;:.i:;lru.:,;i-JiEttli:

tf75,-3f=. q E A a

.i

i P C 8 e 8 a; > A e E

3!

a 8O 60 40 20 0 Contro SA654 1 5O mgJtg rndomethacin 5 _mg/kg eT

iTO

-:

O 60

'$

50 =

E

40

=F

=-,

e

20 e

S

10 O

g

0 CcntroJ SJL654 1 rndomethach 50 mgn(g 5 m9Jk9

(25)

q uJ

i

i!

L;

'!

BgE

SOB a See W loo

Contn)l SA654 1 _Indomelhacin

50 mgJk9 5 Tnykg

Figure 20. 5-HPETE

##?9R#A.Pg

LTB4

_(a),

PGE2

(b)

h'3:tFLTC4D4IE4

(C)

i*&15ik#i-8

SA6541

_OffjB

**1<0.01 vs _{the control} _group

･7i-7

[!1lJ-

ju [-i-[iiL:+,Ir.i/

-/JI{l.:.;:1i

:IkJ-i)-73SAG:-)-ll ,/)J1:))ill.

:)L_i:.TTj.j!_-J}-

tti-.-I.Li-)Ij,;TSiY./)

Sodium _arachidonic _acid (u, Sigma) , SA6541

(*RA##iCe:it)

h'itF)

indomethacin

(Sigma)

kjEU'f=.

ILLi-_llWL-.))!1I./)

_1,1I:Ultf!I..hJ[-i.I.lr!t)[;rf

:i:(II--6-7

_3g#o)#%ICR

_79xk, 12

_{FSFRqO)Pj]P5i9-j}

9/I,

(7:00

8=Je(fT) -F. iiR23iloC

:rJt_+.L-i)lti!"I,_)ti:5;-)-tloo;,:_{=n1--) -=1{tr..hJi}L f=, [L']11riJilTiL-JJl'::I.iL-/i(Iil'lLh (=1i-{L/I._. I:)i'..I.i-.)!E-tJ-L,i>

=T-'[li:i-((j)i-)i.I:::Ii;i:-).LliL-))-hfL /-I._

^A.:.lTiiE+,-i/I/1_ll-_)'r_{:3i.1.J[:-:-E-}_:i:I_JL

ju (80 _Llg/mL, 0.1% _ethanol _saline %.jgiii) 25 _{LIL k} 30 F'--}&o)7j 9 u,>U y.i

<i-Tn.-i/-( rL/_-/Jr_)'lh(_{=rrZLJlti_ii[fJ-}L /'-I. /1J'--IT)i.l:([if':i:(}))tLIJJ''L.Jn:I. )t{LJ,J_{I;_ii1'rTfi2}_Lq IT!J--f.".):=]TiM(JJ-'1'._:

(.)))1't:t.ilk:E.ii-:.I

L -' _-Ii:-k'/Lf=1

-a-1 I/iiLrL..-I.':;;:I:L{L,'i-6 nm1 -)}L', Jr tv f- ,I_> +-Lil-rI-,i-)(--=T

,

[i-'tl'-:I.i-0.5{,I.I,

･J---TT:'-:i JL T,T.)} +JL T' i :E-:- _T)ll 1t=7 _{-: I,tt/'LWmM} 1)i. _{tTT1-.:fa''J-i-fir.-itltll,(-}o.7..-"llJ[fI(=l.1-'J'r[7T_L /-I. LTB I_{:rJti U: PGli=} ilTJ]'iE)[H.I, ]!{r)lt;1--_l'H-fS!.'3()'J):I_][illF..u.'['._:a)lil:['qlLt;.ii-!.I

_L

-' _-=j:-,/(/JE I-_bt-Li-.

//LLlr.I-..I:..l'iI''(;[.<'i-6 nlm _-]iI/I jr

I./ 7,, ;i +-.=1trf:,i-)(-i:I

1

[;-Jtl::L.'iJ1 ]nL =r- I/ / -JL III(=['1'(Tl7TT:L i=.

SA6541 h'

ctUi+ indomethacin 0)32S

:<.HT:I.i.-(rL/_{:/J=E-i.,T:F_}_[]A-I(Llj-nl1.[;j7)J(})_{-ZLi-;i':F_}-I['{Llj-L , ,]1Li1'y).l'{L+-Ir)

I/I _{!=t-LL SAG.I)ill.,I)()}

(26)

mg/kg, b8V'Ehndomethacin 5 mg/kg

k,BSR]2SLf=.

_,#fifR]2ji

1 FSPpq1&, AA 25 pL

'i{.'3()Fi-.Jj-.'1.,))-I1 y I:1_t> _[)i j,i,)flu.-L,-,Ir> I/lll-J'r(.=Jk[)ltii-:lp1.[L /-:. /i)I.[lTIJ:!jH:i.i:J)il7JJJJii'

a)]-i.5.E,yt, AA Jkl),I.iiil'TI2 _[T!J-:If.I,]fi2(I_{lliJ.i::.]l;i-1:I),TiJ3:f:}IIj'{LI)-._{L /I._}

-II _{UJ<)I,**,/9ht-if'} _(MPO) _fE.r&niB[1E

:(;1

[-:I,-:.'L()Ii [[rJ-.LT..ldi/<7=-)j-tii{LTt')il-)j'Lu-L /I_.

r:LIP.I:r,Ii -(_):PGlh _{())ittTJJ';ii'}

9->7p/i,k*%-}+j i

_(i9fflk)

L, 15 /i)FpqL#JLl5}#

(12,000Xg)

kll-3f=.

_t1#i:

i.Ilk)JiI:I(ll"i{1'[liL Ji;tt:'Lq1''/),i-0.1 M I/=t= i _tl'i-7:i+f, I)r1/_{ll,i'(I-iiJ..ii(''i}(pH _{:3.(V..3 lnlJ}:I.iJi;i[('J,'l!LL /I. T..:i

77ik C18 SepPak A 7J>

(Waters its)

8=jBB3gL,

_1%gFg*%#

25%} 9 ) _{-/I,A(#iik7?A}

t#* Lf=. LTB4hlcEU( PGE2 ij:

1%Wg*%E

80%} 9 / _{-)i,A(i*iikToi*tH} L, #n#z.B]k

11J3f=. LTB4hlit>. PGE2 Ei##jt4i7.yk/ *,y _-f,(Amersham

itA)

kjnlTiEJJ1%L

f=. LTB4 jsli U( PGE2 %Ej:fT6]kv

_'f=#A-#R#&

(0.2826cm2)

_nf5*kjBV

_IT#iE

_{L,, pg/cm2}

(I)fL[r1:LLL vLI]i-L /'=.

7Tv'LT.-[I-[L'J.'f1)(.

1-J.)!t:uf.l'.:._Lj1'riq[y).l'(LI)-._LftT:a₎_;I.I.::f

-iI)unnL!tt (})iS_{Lr,_:lt.iL,1'}1t.;!',i:_,i-)Hi_1-L1.rJ!_1=)T-L f=_.

I-]i--.I.L-.3.i_;;I..J,:il

i-AAW

AA.-.-rihS-rL/ I/Jl-.)'r:LL:)Ill:-_LIL;rJtE[ i:'/1iI_{I'l:.lit.I.it(llL'l}_(.I_i-J--I)-i.''_SAG..-)-lla )n-:)[],iJFigure 21 :I ,J-I-)

-SA65-ll 50 _111ff/kg. 2 rrT1_,j)]'<LIj-L'_{i. 1u} iiil'1J-_(irT!j-,It"t]lil.})/1J'.I_{I'JilitI1.itllltrJ}

;iJ-:;I:J'j.i-L-;;::J'l'.)(I-(J-..i-(.=11iJ-''fiu

L -/'=. .

)J-. idomethacin I:i_(bt.-i-[iiE+,, r') I/,1[-)'Llillli(.I;rJ1:[ }3/i)I-IITIJfit.i..i[r.[H=i-J-L -i, _{!;:.-I;:;lr!T:J1.}_Jk 8f5 tj:75,J3f=.

(27)

.∼ ∼ E IO C -ET a a a :2T ∼ U ∼ 0 A = C S ∼ & A_ U E 12 10 8 6 4 2 0

Control SA654 1 fndomethacin

50 _mgrkg 5_m9jkg

Figure 21. ju

##?

9R#*Pg#**i2ifq8=#i-8

SA6541

_a)ffFB

*RO.05 vs the control _group

ju.iTi lET-(r> I/Jl- 1lh:)i:)Ill:::.I;r,I:[ i:) LTRI J't.L Ult PGth _it.Tit).ri.II-_

R.(.I'(-J--.)-73SAG.5i1 (})(1-:)[1

<1-Figure 22 8=,j%+. SA6541 50 _mgntg _{n]32SEj: AA.a#J} 30 LAli&n LTB4 0)tB-k#=+SBb

8=E%8=qnSr]

Lf=AS, PGE2

_{0)tfLi=Ej:/}B*4k&EfS}

tii),J3 f=. _-jf, indomethacin Ej:AA .I,-Ti-SET-,-,r>7Jr )'r:}Ji,I.tE:.I;l''t(+,:3 PGJj:u (J) [-_I,l.-,i-a''l,'i.:i-[-;-;::I'1'J!=

[J-,.ll(=J-)iJ.liu

L -/-I/Jt;,TJ'LIBl(J) J-.';H=

E:i!;:.;:'!.i.I.:,;i.)iIttt.-_I-JLt))).-) i'=._

q E 80

a7.

∼ S GO

i

50 C>

g

4O U a; 30 5 20 C 3; 1O tl

S

o SA6541 50 _mg/k9 Tndomethacin 5 _m9rkg eT

i

1400

-:.200

.SZ 5 lew C

S

800

S

600 Lu

e

4CO C a 200 ∼ 0

i:E

0

Control SAC.54 1 lndcmethacin 50 _mg.'kg _{5 mykg}

Figure 22.

_ju##?9R#ALPg

LTB4

(a)

h'j:tFPGE2

(b) *&t5i8=#i-8

SA6541

a)fFjq

**RO.01 vs _{the control} _group

:-(i8 i.1i_-;;I..I,:!Il-t _{-r'.-+J':,:.i}

1)A -_(I)1--3I;it-J-)I).tt)_;):(J)i::)FLU:lTI,i).)I;'f'J'-I_i:,;iif="

1. SA6541 Ej:5&hifLTA4H

_FRS#vC)bJ3

f=(%/I,f ,y I, LTA4H 8=#+a IC50158j:270 _nM, Figure _12).

(28)

2. T _{I) :T-I,tt>['1T'i-/I).i:)}_;-)-.T)

,1-,:1(-t> I)l1--t}:I i -) -i, LTAl /)I;/Iil)t-:)-73/)I;.JLLJ)[tJrI"J](1=vet(i})73

5-HPImIM)l!{r)i.i i:_)pfI::t i) _L'1lJ3_I

I;'(::_'I:_._L/i)I.[IrlJ:I,I.=1tllT,'J,JiJ/I:_L: i= (Figure 13, Figure 14

h'

cttF>Figure 15). -k, 5-HPETE 0)jkPq.tE#+7?1j:7 5? * TI>gEuo)jkPq.a#JE=tEJ<T PGE2 Ej:E2 i AJ 2j1'F&fE5nti75,J3f=

(Figure

Rob. Figure 22b). Lf=ASJ3T, 5-HPETE

.iFi-1E)i)..t=j':))i-I--:7-1JL (=J)Ii1 -i/ _I-i_5- _I)

/-I::1--t> Jr1---L;1t.:LJlll;lip::1//);,)I;_{lr._:_LIL(}I_Ltf','L..I.:rJ{i-lil)i'C }5._L.JIL'i

I:,;i.5L_;h,/I.,

3. SA65il E-i)tT:F.L]A-!<LI)--:Ii:-).i,-,-, r> I/,

_{5-HPFLTri.;.-[iie1!{)I::)I:)i:E-;/-:Ill}

_(.I;rJL:r_{}3 LT13} I_{I;lJ',-:'J:jjt}

j=tJilif*fRfRFi.IFAkUpSuLf= (Figure 19 h'cEU( Figure

_20a)

AS, LTC4/D4/E4FA&8=tj:

I.;:.i:.L!r.:,L I_Lt/I)- /I (Figure 20c). L f=/Jt;-,-L/. 5- I)_,1-,_:I--i. Lil---Lilt.:Ll')Ll1''/)(I-1JtL1-i, _LTR1

7)u1[-[fI)i.1itlt,.1tlll.'H=tr,:iIB-.=t.i',)i3I._L

/.)I;,]-.-[]7'f=L-1)i-/I_.

･1. 7J I) :H1> [ll-fi.,J)rlLJ,)LlilrfJ-(.IJ;-A -i/ LTI31. PGIh );-l'=:!I:_;rJti -i)'t/i)I-[T.I:.1(it.i.Lll[,lJ(Figure 16.

Figure 17, Figure 18 jb-iVhFigure 22a) bSiBhb6YLf=. Lf=75SJ3T, 77*

T<'JgE-.:.-Tilc1,t{A-:).:ii:t- :/--ttJL-L'TtE-iL;_1-.'I;E'dLtT =}:H,t1i [1-fi.(L-.iJ'u4Jy)/)I;I;.I:JI-7:.)-jL I.t ).iI;Jf(,:I,>:i;iL1;h f=.

.5. SA65L11 [-i:T:F.[=]J'k-Ij-:I_{_i:}

-' -i.-,-, _r> I/I_ju.:;[i_{lE).l{JL:.i:3:--E-:i:JL}_{:I;rJt:[ }3 LTD} _I

);'1'---:/IJ-'t.i

Ul7L(-q]fRfxFi,n3BkqpSTILf= (Figure 21 331i tJ: Figure 22a) bS, PGE2 aFA4=E=Ej:/ad@BLtj:75'J?

/I (Figure 22b)" _{L -/-I/J1;}-) -LI. _1u (L.,;lu4:y)

-7t_(.f=(I:l'JLL1-LI

l[,

LTB 1/)u1)I-II'l:J;it.i.ill,'H=.r.:lib-Lit

.i;,}3I i- 7J)I;,]-;I17'f=L-1,.)L/I/)I;. SA65-ll :I.L }L/ll'-Jf'l:Jftlt.i.tllrL[JJljJ''[;-u)'Jt;I;i_{_1.'/Lit.I-i/JLI}i

1 (Figure ₂₁₎

I._L )'J. i-,. (ui,J)_ju (L-.i]'u1'/)lr,/L1'-[ITf:1l-it.i.tll'.'.H=Jr._:Lhl[vL'7t.(-,,}3 I _LL _{).)I;A([::I;.,:-i}i:IiL)3._

6. Figure 22b

_{0)BE,fiEIB?5,6,}

SA6541 Ej:4,FjS.%E=jBU'f=jq% _{(50 mg/kg)} 8=h'VITEi-J

y r1

-,1-:T-t>..I)l1--tit(I!;:-!l!..li;L fLtL.._Li.(I'iJ,i:I:1_,h )i/I/;.5-HPJi:TE ii:_l'1J.:IL ,-3PGlj:L}I;I'=:/I=_I-i

i-)iJilil)-I)-}3(Figure Rob)" 1'.(lJ-l'J:1-j-hi-J-:=JJtLrL, _LTD ₁

,J)[kJJ,).Lti:.i-H.)'Jt;PGJh i;I(::'J:_'i-.)I-=<ltdI

-.)-I i k#iBhLrCh' 9 , SA6541 E=i-3T LTB4 0)FAfEbSUpSrJ

5nf=,#Btt

LT PGE2FA

'I:.i,i-li1.'fi[Ji3_;1i/-I._LJ1(::I:,:uh;

,h 73_{_}

:Ti-9 [iTi j'J..-.;-lTii

LTD I;Jt;FI{I.::)I_:3i_,J)1-:)-:i_I/i(-[(T]:J:it.1tfl].lH_I:TJLL1pL,. _{r,_:.ii[fLt}_fJ(LL:frJ

,i-Lli-fr)--I.1r i.)ti,]-.-l17JLpli-.;iL-/I_{_}SAG{)-ll

(29)

A

2

i

#Rt=Jf

= h U_Iy

_{A4hph(Jt3###FR%#}

_SA6541

_a)35*W&UfFR

･'Ti-1

l'Llll-

I1-)r-Ljt(J)L-1n'J

I:)I-):[,iLTAIH I;LLJ.;..::.i.fi-SA65L11 E-i50 _{mg/kt,u lJ.}_{I-_.})X:F_}H JJ(Lil-:IJ:-' -i,. -( rL/

_{-/JI{).:.-..':3i}

,7_-i.JiJ.'I;-lJ

1-}IJI iJ

_{i.,'Ti-

1.1:'J-:(.=IJtL,-i/

,]-.-L i=.. JLl I -Lit SA65i1 (j)i-;[1-LR:)LI':-_n-:)1H=/)i,-i.-( ,i)_{711ui.1T:--t--:i:}

/i,kjqU IT*il Lf=.

#2BS

SA6541

_{0)#BlfjE7oT=77JI}

) y91'

･1<(L:i'=}r,/)(J)1;R!i'::(I rT']Ij.-)-}LI.[ffEIJr:_/Jt;,1-.-Ilif5;h --L,L JJ Jri.JtF-(,1=J(,(ul,j)tTi.T:,.I.i:-(Ii-J-I)

-7y!;:!:.L'i;;i-1L!7?

i4Lf=

(Cerep

8=TjfBhFf). _,#Btk Table 1 8=,j%1.

*BiljfiAEj:i;RfnJt

A- 9 Thb8.

JriJiFLHL-JJ)#t-[LT..'TT'iE:_{I.2.77r1;q-tT:}r!,1,--r1(]Jj'(lrlJ-1JliI) -)j> I,1t:I.(tJ.1-7t''SAG.,I)i1 (;3 LLM) lr,..():-H!J-:,J);i':.'I:

%PRS-*

(%)

k#tH.

#mB&#jiSbt:7_,y-t=j

(m1)

:*uiBklfjnj:*7o+j _-Jy

(1

LtM) k 1 i Lf=iB%o) SA6541

(3 pM) 0)*uiBko)ff1%

(%)

, LLeWlfREj:*7o9-1 -Jy (30 nM) k 1 i Lf=tB%o) SA6541

(3 uM)

jf#F$0)LLkWo)frJ%

(%) kSEH.

##7 _y*j

(COX1,

COX2, 12-1ipoxygenase) :*#8=ct875i* TIy#uA,61Leimqh

fEhEBZFS8=jb'818 SA6541 _{(3 pM)}

_##F$0)FRf*

(o/.)

kSEH.

Table 1. SA6541 O In vitro

#%3tB9a3jB#A

Receptor/Ion Channel/Enzyme % or % Inhibition RefTerence Compound IC50 or EC50 _of

ofControl at3 LLM Reference Compound (nM)

Cannabinoid CBl _(human) Cannabinoid CB2 (human)

Histamine Hl _(human) Histamine H2 _(human)

Leukotrienes BLT1 _{(LTB4) (human)} Leukotrienes CysLT I_{(LTD4) (human)} Muscarinic M1 _(human)

Muscarinic M2 (human) Muscarinic M3 _(human) Neurokinin NK1 _(human) Opioid kappa _{(KOP) (rat)} Opioid mu _{(MOP) (human)}

Prostanoid EP1 _(human) Prostanoid EP4 _(human)

Prostanoid TP (TXA2/PGH2) (human) Proteinase-activated PAR2 (human) Serotonin 5-HT I A _(human) Serotonin 5-HT2A _(human) Serotonin 5-HT3 _(human) Vanilloid VR1 (human) Vanilloid VR1 _(human) COX1 _(human) COX2 _(human) 1 2-Lipoxygenase _(human) -3 CP55940 -7 WIN552 12-2 -6 Pyri1amine 0 Cimetidine -5 LTB4 -2 LTD4 -3 Pirenzepine -13 Methoctramine -6 4-DAMP 1.1 4.3 6.6 230 0.5 1 16 81 0.7 -36 [Sar9,Met(02)11]-SP 0.28 -4 U50488 2 DAMGO -16 PGE2 12 PGE2 8 U44069 8 S LIGRL-NH2 23 8-OH-DPAT -I5 Ketanserin -12 MDL72222 -3 Capsaicin -8 Capsazepine 3 9 Die lofTenac 54 NS398 9 NDGA 27

(30)

LTB4: 1eukotriene B4; LTD4: 1eukotriene D4; TXA2: thromboxane A2; PGH2:

prostaglandin

H2; COX: cyclooxygenase; 4-DAMP:

4-diphenylacetoxy-N-methylpiperidine _methiodide; SP: _substance P; DAMGO:

[D-Ala2,

N-MePhe4,

_{Gly-o1]-enkephalin;}

8-OI1-DPAT:

_{8-hydroxy-2-(di-n-propylamino)tetralin;}

NDGA: _{nordihydroguaiaretic} _acid.

SA65il E-i3

LLM a)ilil'l]ii=:I;rJtL1-i/ _,i.// L-7_-)-lT-,iJf1--t}l-2.J)iL-.JII:_,i-_{{)-1l.!"qr)i'fi-u}L /I).)I;Jut

a _)J:77i:Ji;i(i;J('I"i.tJ.:,tL:-,∼ )_{I.;!-I;l!:I.+i}_IIL _{i: /u}

LLttiL),(t!)i:,,hjLt)I).-, 7=,

tijs,, f}L{,, h

_IJiA4H

8=#+8 IC5.1#Ej=0.27 _pM

(#1

i) _-C)babs, E h LTA.H

t=#+18 IC501#Ej: 0.025

LIM (Scottish Biomedical Ltd.E=TjfBhjf) ThbJ3 f=.

:-(il..3[[rlLSA65-u

lT:F_IIJ'+ilJ--fii.(})T1'tLi!;i-JII;'l"i)riflll':-.r:/I

･'[il1.I;I.-:(=J'tL.vL,.SAG.,-)i1 (-i.r-)()1ng/kgr. 2 IL.I,J)3:F_I]J1{LLi--Lit5-HPETE.J;:-1E1!{].:r:..i)1/1J'.Lf'lJ:

'IxFi.n3qk E5Ea"15%JAI8=qpSr]L f= (Figure 19)AS,LTB4 F%fEUpSrJ Ej:5%/ekE7?1j:tj:A,J3 f=(Figure 20).

--rl-I -LTtt f;tfiJi:):I-::n-:lTl{iJ1-!hi:I.f-.)-}3(I.i-,I-/Ii). SA65-ll ,J)j':F_II_{l''Lij-.Tl.i:,1J}100 _1nEf/kg.-L.L _{it(:_F:I}I

A:!{Liiiiit(J)_-,I-{1i:fL:(iL-lI'lL!)i!-[_II

I(I.i"i).:r'JII'::T:Y,'i-1L!;-I.i--J-L /I_.

I-)I-.I.j:.;.I.J/tii-:

I.Li-)I).jlSi:llu

SA6541

(@jeA##ie;Ail)

kjnlf=.

I.Li-)I)J7--))1_{1yJ't I:(')lt]{!.r1}=r'[-i-.I.llttll;'i-:E f'f:

5-63A#0)#%ICR?9Rk,

12

_{FSFpqO)PBP5+-1}

9/i, (7:00 8=.h*T) f, fiB_23tloC

:[JtE-i":ti.flLl,_)ti:55 _i=10t!;.:

I _n1-/'

-L1!',h'iL /I. ['-.1)I;.l-J_i!r.I-rL:LL7J(I-iL][lJ.r=ii-{L -/-I_. I)i:.I.i.;.!t-i7,,,:.-)(-.'it5:]tSiq!')J

lf/)I)i'-.[.i:j!(f;i7l:[I!_F<{

-lil:

(I)_,I(.ill._;i-f'J7:-L-I)i--))h-L -/-I.

SA6541W

SA6541 (100mgnig)

&R]3t%4&

o.5t 1, 2, 4 jslctUi.6 FSFRqO)79xhi#k/LJ<U

_J%

I([t> I)i _{tJvL1-t[1-ll-(}L /I._.[lt-:1l:i('Sit.lLii-J(j;,.).i=jii-JL.:I i i)_{r1'I1.3tI!-,i-}_'J)Il:iir;_L. _ImTA

_{().:-(T--_}

_[I. _{li".J1-:'i.f"i-:'iJ}

LrJ[i1=:.I-I-LqO C -L11'1''().:L /I. SA65i1 1h5lii.[[TIJT3J-ri:[itf1(l'/:(i=I/ [J --., r,

I/ i;iI-/JL-JE-_ltf'L:'J)7-)T-:ili;;llllrT:

(LCMS) kjn _lTiRr]iE'+Lf=o

:)i'-.[.1-.3∼l;:..I,:ll

(31)

SA6541 (loo _Tn!hr/k{:);I:pF_[]l'<LI)--fi2,I)_{[h_;LIL-)}II.,lTttJ''i-]1(:-_f(!''iJFigure 23 (.I_,)-.--.)--X:lS_L]A-'<L/)--₃₀ _'')

4&nh#,J'S*#JiEj:

684nM Thb 9 ,

#2

B5o),#j% (Table

1)

k5%L8:+8

i, SA6541 (100

mLdkg) 3:lf_II1'{LIj--Lit.;-.L!..t!,_tt,3L_,-3l:i-iq!f tat:]--.j.I.i-n'JiFL'LTAtH I:[L:.it-:(I ).Ii--)₍ l[,_{a)._L}

,5-

fLI:',iLil.: i, E= TTT e ,9 ∼

B

E= * U E= O O ∼ E d ∼ L loco 8W 6W 2 4

Tlr7le after admlnLstratJon (h1

Figure 23. SA6541

_{(100 mgAg)}

_{#n#%@e)A#q)#JEii*Lf##}

A

4

_BB

5-HPETE -#hk7

9x1S*ff'}t,0)1fA

I_

lit-.I!)-L3!)J-_ti1-..

I.i-)fl3]tL:1ly)

(5-HPETE,

Biomo1)

kjn 'f=.

I.i-Ill_iP-J)q:,/)_{JJt E Ultl{i,.-hJ}

).-i.'lll!L)i;ti-.'iyI:

5-6

_iq#o)#.EEICR

_79xk, 12 _{FSPpqO)PBF5i9-1} 9)i, (7:00 8=JeifT) f, fiR23tloC

;rJtt Ulti[l-l1._)_tli:-55-i 10'!t''(=n''-)-i/f![.1TJ[-L -F=..[-l'l)P-J_1{trlJl::I.._L_-/I(t-tI')rh != /)--fL/I..']i:lT).;.!E-ti,i-)(-.'it!-,).1iul-))

ry)I-)i'-.[lj,'c((;i7ln!/FL{n

_:_:?

a )_{/((.;-.L!/Ji.}'Tt.)'--_-II-)i'-I)kL 7=

_,

5-HPETE _{idi3E7 9 x4%4jE%f'/i,}

jS& 2 _{PBn=?PRO)j[jb#kB%%Lf=.} _5-HPETE (0.1% _ethanol _saline #igaf&) 50 _pL

･Ji-27 i)lt- _,lltj3l.a)-(_,I//T=!_t> T)_i.>lt;iJJfJL1-i/-( Tt/_{-/1't-,:;1;:I)[{[)1.lt}i:_l'1J.+}'JI i: (.Ii -' _{-i-1-ui,):LIL:.).i}

T,i,-,Ji::Tl--JkLI-I- -.-I

r>:/I