イソプレノイド代謝酵素を標的とする制癌薬開発の基礎研究

(1)

Title

イソプレノイド代謝酵素を標的とする制癌薬開発の基礎研

_{究( 内容の要旨(Summary) )}

Author(s)

遠藤, 智史

Report No.(Doctoral

Degree)

博士(薬科学) 連創博甲第5号

Issue Date

2011-03-25

Type

博士論文

Version

none

URL

http://hdl.handle.net/20.500.12099/36295

※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)

ft _:I(. _{( -1= jlt,;}) 'li-/:

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a) _{r'rt'_}_3.lil I:I=I_.'[_llL-i)-.Iff-il.

q=

1& 1-i.

-t5-EJ ITJ.

ij1]--

4(

'3,i

Ji.fJ8

3L-Lgi-E]

I:;-I:(Ill:.;']T[7i- '1i:;.l!J'.A-:ii

;ii )J'.)..i Tin.]LL ([Ili-1'.LL).I..)

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5

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tM

A f-:).

_'l-;I

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(Fundamental _research for dcvclopment _{of antitumor} agents targeting for

isoprenoid-metabolizing

_enzymcs)

(j=ZE) #lii _,# Jrli _::.:,::

rtli

(BrJ2kij:)#l2 PT qq lLE a ((.';lJ4f)im'i. ]E H i je

.;-[;i7A-r)']'ifF'a)-iH--a

lf.1.].Jlji)JB'L1-[l=,-filliU)---T_[TiT.LtD,-i)7 L,I _{JL.[kE:iJf_t/J}L{-.'1't_-}_GYP _hI.:ET)=r,.[_-i_y >/i }}_'L-'i'_(Ras. _Rho. _nab ,illt1 (I?2y<

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,

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T:$11-+l3(hu)y J/i I/

_JL-lil

i.a) 1:[LIT_n-:JrH=.i'3tU _ITCir,.:-i)Act.L<T)25L.

iE*, 1

_{i,=/i,1EO)&ffTC>b}

8 farnesyl _{pyrophosphate}

(FPP)

i _{geranylgeranyl} _{pyrophosphate}

(GGPP)

8=hnkT, %n%>YLnARU J&ulk8=i: D 4IBk+8 farnesol

(FOH)

i _{geranylgeraniol}

(GGOH)

8=i

84#kf2fE4hfELr&bS*RE5hT5f=. 3:f=, FOH 2= GGOH 8=Ei*+--Pti=ct D t=oT] L1>B&I4=J-i+/i,

J<--}5n8#ffifB6A';,

FOH i GGOH n%ulbI4=-e>b8 FAL i GGAL 8=Ei7pt/I/i,1t:9

>/,o9#0)##

nt))I(q.'1I-;I._it)

_{!-L-_J'q+7JSS_U;i)'JVl.:-(Ell-,5L.}

L ;J,L. ILlu)ft:.-;WC=1U,TE-i. FOH i GGOH fJt;7'JL.j='E 1,11= (FAL i GGAL) k#fEET*}L,7Ji-yg&IS (FA i GGA) 8=FAblk5YLaggEB8b1;Pj] 6A,C=5nTUla75S, LR%

+8##i=-3V'-Ctj:je7:=TL^ii=ASPB SalTV'ifV' (Fig.1).

A=W5Fc-elj:, 9 ,y

T7i!B_5&&U(Tf%*#WB5TeikFf]VIT,

7/i,=-/L,I4= (FOH, GGOH) l7/i,j='t= f<1I4=-A .JLjit>(1.:i(1;v)rL-I-H1

((t.i-410Lh

@)

:=TA1,i?,3

##8=hRk-C, 7}Vft

T<'1ISb,67/i,=-)i,I4=JlniiAitn#1t;aJ#fEEE8(Lt#@)

i i

a)AhiE## (j y7oV=)V7/Vflt= TIikJAjt

##) _nF]fk#*T=. 3:f=, jy7oL,I

}v7 /i,flt= T<'>iiAjt#if5jSa)

_PRS#J

tj:7p i, =}L,

(kU) _I:AHLL=,+iJ(fFl]i;.BJ-lJ.:i Tl-73 irri-l,i.'1-.-u1.A':uF

{l'Ltii(Itit

u _{5 ,3}

_{tm,:{3,:Sis,ilf=}

I

LL.71,it_).kf].-i-.i-8t=

7,a) _LW;.J,.;-a) _[F]Jii: lh

ILL_a) _I:Fl7i#J_a)

_f'iEJ,i,;I

,->i-I(iI?

Fa,nes.I

gi

Fa,nesal

@i

Farnes.ic _acid

(FOH)

75

(FAL) (FA)

Ace.y.C.A---Flip....F.Ia;e

I

Gipp...a.;:s:renlylated

proteins 'Ras,ho, Rab

etc.'

lT

nl

I

@

Ge,any.ge,ani.I

&Ge,anylge,anial

_ Ga,anylge,an.ic

(GGOH)

75

(GGAL) acid (GGA)

Fig.1. Metabolic _pathway _ofFOH _and GGOH

1. 9 ,y I,ib'itFt= T,8=h'818j y7pL,)/

f<1Le###o)rqE

? I/ I,:=J-J.:+I)

(t-.-?'JKi)a)lJM(i,f.frFF;i,;-v)[F]Jlii'

: _{7FI;#,){,:-i.T,-.ri,+I}-5ElilfTl-73.inI'::LJt'

L7ki-L,3 U = > t=1t-I-i I,FW

#kj7]V,T FOH i GGOH

_{8=#+8jiFS4&ktE4&L,f=#A,}

2 #0) alcohol dehydrogenase 7j )9i+-1

A (ADHl i ADH7)

AS&vlhd!#/3h*k/j%

Lf=. 7 _,y

_I,gA#ToEL

_{A=##fE1Eiqi%t*J{3}8=#hb}

6h,

-1L- _a) nL I_')

)'&/JJWT-

/I;ll)-.-.'iilBf;ii:-

I) T:rl7i#u I;i!H,2.r_Ill.bkUqI][(-1L-)jfI.i,-.[J+i.i:u]Irarfll(-L1.-?A.r'):-.L!):::iiJH:-)JJ r-LiJ)JJt,)--=tEi IIDH 1.

(3)

-9-[[TILy_{))lfJ-TE∼:i}ADH7 _7TJt;A-I(i-..ilJfu)j:--_LA(/iLll5L-A(:,),i-[lW-:a.,:--=t(-i-,)>3I _i

7)I;7T.-LTtiii-I-.

? ty I,:=JJ. (+ <)(t..iL'Jf(2)a)tl-7'f(i:tl--.Y:ii:-a) ri7J7ii': --/I:I I,

/1:uh''iliku)i:tlllJL!I)}[bJ-7t:i,1<LlhlS-:).i:.tlt..fT:I:_fi

NAD-(/(-((:.TJI.:

Ti 9 t] _)-L8=-iBhb _6n,aldehyde _{dehydrogenase} _7/ _y+.JI Ln ALDH3A2 _rChb8

_i,j%@5YLf=.

i.I/ _{1,:=J3t.I+,I)}rL-.iqfr3)V)jSt.;.jt(pip:i,ii:.a) rm_I,ii'..1=Lif=-)..:.71.;T;-LIJI:.fiNADPH (i((ilJ[i-=t_{JJlh-, [rt).-);fJ!,i.a)}TTt1"..:i.ri @J{5}E=6 <. i a)F'/Vbi&/hW.

FF')iA##-nPRB#JhLIG%Tk

i 3Ef4=jiFS#75, 6 j=##Ej: _aldo-keto _reductase

(AKR) 77 _{-I i) -0)AKRIC15} _rC.b8= tA;Pj]6A,8=iiJ?f=.

nJ_i, 5i,y f,8=jb'Lla FOH i GGOH 0)A/I,ji:y6*H=Jlnlti4J##js'i:tFiPjiFS8=Bg%+8iEjt## krqjtJ+L,flo =nikmjtTRFSEj:. /JI4bRI-T4!A+ibV)

,

;p-31t/I/i,Ike)&KT.b8

ttoT] U

>#f8=2g]5n

8 FOH i GGOH

_{0)kqnFeiPqmE2k-3.T.Jm+7DiTi#tJ:#r1-t-a4J#B8}

i:

_j%k

6a18. 5i.y h 0) AKRIC15 8=#FS +8 t= f,a)j y7pL,I/L,7/L,j1+t= T<%gtit#.a.:-kL-LH+FIB, AKR 77 _{-I U} _-8=hi+8#kO) U = _ytt1'+

> h##8=i 8 FAL i GGAL

_{iEfjt##-f-I;-11t&4*.Li-A+Lf=.}

_%0)a-A1

7/V T<+-Rigit##5(gl9 >/<9 Tob8 AmlB10

_{ASi&b,i)u-*BqL=Pq/}

y7t/I/i,7/L,ji+t= TIi:igit+8 I _{i bath} L,f=.

2. t= I,_nj y 7o L,I/i,7/i,j:' t=

_{T<'iG-jtBi-a,:-rSAS#ja)JHi',qS'}

t= f,_a)j y7L,=}L,7/Vjlt' t=

_{f<tiG-jt#gif-C.b8}

AKRIB10 Ej:,nfTHi7,

B6#,

j<B%*ii

2t11vehba;#%+ ･=i)

)LF7-

i)

-Tt_Lbu. _{1[iLiU)jiLJii+--a)}I-i-i-S-.+t)i:t!lL--_L7k

,1t, )!.A;a):)I,I:)i:r-JI

LiU(tib* (=&3t (+ ,3i't:TrL i,TTiNtJht-I-LL.

-rS-k6alTV,8. %=T., ICM _{7pT=9>t,Lkj7]UITAsinexitnlE%4h9/715i} U

-(5

_7flk%qh)

A,6A= tva,;-iEi;--[t][.lf11'i3LfJ).irTf_'tJj-17,L;I-_(I;E,I.:(5T+73 _(i:

RA4bi:7, // U -I > // L -/=o iE;-.r'-IiiI,'i''(;_I:4X-):E-i a) 1.llii-_[fma-) 7, = 7 a)[E:;L_{1(i:(-T4bU)I)}Jib. _{lui-tfllTU)TSrl7T;(E.?4bu)f1.Ii3ii;-}i

_j[(_tJ:i), A-I:7TfJtJL16 (i:

[?gb,+i-iiCIRL

-1=tl%-;fi:-a)r5IIJ=

fEJr*kh&i4L,f=.

%n,#A,

9 t= }

_{J#k#+81k%4hbS}

AKRIB10

_{0)5&j]tffErnPRS#uTC>ba}

I i: k

3attiL,flo *Te>b Fig.2 t='j%+1k%4h CI

_{Ej:BE*nO)i&b5&j]tmRS#Jct}

9 b 5 l%nJ

t5&VIPR%5ilaAk,j%

Lf= (IC50= 6

nM). if=, CI Ej:f%4:5#.mn5Jikji]U'f=9S.5&rChb AKRIB10 n-1 y7t/=)L,7/i,flt= FhiiAjtfEJr*kPR% L, (IC50=0.8 _pM), #tmB&V

HO J< )L _Yet+u _--LTa)

_I(--I-

_,i)).r]!_7TJI;_{T]J((-::ii-.i).}

LTtilf=o

1=t[)FrLTtfL /-I--_Pl!JlillE')(J(=i-rf_tiitfJ:_J{ 1,/7' L, /

-( T,ltd)ft.--AIPiS--lit:.a) F7]'ii'(I))[]i I. -( 1/

7pL,I)i,7/i,fhI= T<''iiAjt##8=i

8jL,=}L,1b&#ni54=

i v1 5

kM&fB#8=?q

Fig. 2. Structure _ofCl

b8#ii&fSkPHPB

L,flo 3:f=, t= I,nj ylv=/v7/i,flt=

T<1ike9jt##0)5&j]iiPRS#uk

in silico R9 I)-I > I/:I.i u i-LEOL t=,i I

,it()a)[Tl)FrL;'J.:!fi-Ei_'Ji(1=r)i7- i,I- JL,(i:iJ.I.]L!W4<;ulViU)[r'J.'tI-Ly].f31i:U(f'frL L1[fJ1]f;B

rJ).-.

_{a).lilJ:kL.L.Li5F5U)}

_[l[I1

-1E:I -? tL1/-)1J3i -,L-.I.i i:)_,i17J.,

.f[:i7_i-Jd;-TtT

i:,I.-lnta) [_lBa

lf.1.1.y-lfT2J)BJf'_':r(l}f:i.:1'FV)-fG-Li.Li;D7J7 L,I )L [k[i. ftE;-)-(-lITJl-GYP i].:(-T49y > /,?

JL--'i-(I

EL

< ;;L'.d)(_)i,i.T,T!nl.]y >/, // _{'Lf1-I);I:u)JEi1)F!,w)T.-/SfT-ttJId!v)}_y > /J/ I.frlrLL.u) FR -L1-_n=)fl:=J-J'Llv=-_LT,:_iEI7tL{T)73c. LFl'f-.7- L,=JL,(EU)

i:E-ffvelb8

farnesyl _{pyrophosphate} (FPP) _{i geranylgeranyl} _{pyrophosphate}

(GGPP)

8=hDkT, %i1

%hnnAR U >gEuJk8=i: 9 fEBk+8 farneso1 (FOH) i _{geranylgeranio1} (GGOH) 8=i 84#k t2fE4bff

Jr&AS*Rf5YLT%f=. 3:f=, FOH i GGOH i=Ej:*+-eE=i: 9 ttoT= U

>g*[q=J-LL-i

1)i,J<--}SYL8WfE

B&AS, FOH 2=GGOH _{a)&ult:I4=-Chba} FAL i GGAL i=Ej:7oL,I/Vlk5

>/lO9#n##By7JiWf8=j=

9 4=

(4)

-10-Bit+8k;FEEH8bS1--fTf+8. L,A, L,, %0)1tJan=-)V ,TEj:, FOH i GGOH _751;7/i,flt= fII* (FAL i _GGAL)

･L(i:iS.I

;]J )L _Tlit_>[.iff(I;(FA LL GGA) (=tl:'![t:ii- it ,-J3':_lI{_UIIvJtH1)1I3)I),:Iis

,-1.i-i/L

,

,3/Jt;. r#[.:i-i)--1-73Llf:}ti:-(I-?itT Ej:L=f='+//JtC=[q,'I!.[yl5 arc Ul_fL>_UI.

*w.gE-e.Ei. 9.y T,,1:tLki&-&Ui)i;I;fi(ik:u"J5TiB&jf]UIT, 7/L,=-/i,l4= (FOH. GGOH) _)7}L,j:tl= T<14=-A /i,7Jf

>BIo)1-t-;;Alt=F#-lb

7D_LjS#L=))[J_kT, _7/Vj=l t= 1,'14=fp6 7/i, =

-/i,[4=JlniZ-jtO)iWlt=##i38

i i ∼

V]d!hRt#;i,:-

(/ y 1 L,I/L,7'/L,j=1 t=

_{rliE;-jti*;f,:-)}

_{a)r7i]k'&;i5tbf=TV} _'8. 3:f=, / y 7p L,=}L,7/i,j=It= T'+ iEijt#-a:-a)rSA2;3;uEj:I1 _{L,I/I,Jt:0)PRT&11=)W13illl(:i}

+7Dkrrlfllf;-uh{#L={j:

9 5 8 i

#2g#

5 hf= I i A}6 ,

x-tt.fFS+a t= 1,

0)#*i:.0)

r6]E i i a)

r5AS#[JO)r7iEJ,q,3'

7u7=I? _rCU'73.

rP

-i7-[')I-4;-EL

/J=f!ii!J=,Y=bW=iE-5giJi>/y 7p L, _{/ /} _{I,tln1-t.=AtfLW;f,:-a)rT1]rEL=))[1}kT , / y 1 L,I/i,7/Vj:1 t= T<I3kJ4

5t#.*i:-t=

_cta 7p

_{L,=/i,JkXJfif1-ni54I}

i V, ₅

1b-kWJ/dJIF7hEtE=fi!:]b

7D

jrrfT1%J})I:&i-L!-FTB

L/CU I _8o _{3: f=,} _t= _I,n/

y -/ L,I/L,7/i,j=1t=

_{T<'l3ArrTljt##n5&j]fiPRS#lJi:}

)'D _S)'1)'coA // U _-=y//L= i 9 RLEtiL/CU18. _=n E) a) _{7)]r.,7E;.IJ=3li-Ei,}_{/i:_}(1=r)ll7 t'I )L [k.-.:),TJIWBBtIILiU)Iq:f!.Fy].i,-JtJ:U'h.UrL i.,_[1I)Il_t1;BJ'J.:0)

rJli-ulll,1'iaTiU)r3r.13E:I-? ,lL1

/J/;73 i -P)-i 6 YL8.

a.?T**n5a.i.

i#1EtJ:#u15#f?7fl#1%f=d)0)#j3]fd:i;.iTt&L).

TSi;&XtLTlnTfi&%tO)i

SuWLT=.

iIi-.a''!1i-,5t.F.59!;F.I.--i4ta) A(a

iBjJ'ltiLt

V) _{,-utf;I-_.;-l:i7}_I a) _{=i7_i1[TL;A-I;}/nt

Ei!.i;-1!1'_H- i ,i-tHIfiiI'i:il.i:(I_{/1t:Lilt;j1,L7f}a) 2 i17'-;

_LFr][T7,uE7la) 1 n1'fia) ii--1-3 all"iU).i-L:i7-E'(I

I,E-1t< i,U)-=t

Li;T)i). -1<.i-I:i7-i',J)I;I:;.i(ir.i'[:iT-A-LJL T'-,tiF5qi-,-ilf=r)r7;i,-'r

1-T+

,TDI _i.,lrn'I:'l-;i-,P.L flo

{u=. '1W,AJ'/i-:E=J-J.L_,I

/I-;.'=flfTi-.;-[:i7i-U)

L),JJ,{Ti:I[iL.:i-T7D_i-[.Ii']:._-LrtLli?J{,

, F1 /

1Jjlri;jS,

*#f4qt.

#f&1t-d1.

L7}+i4a?.

B;#RE]RLni+=&i+;'Ltj:E%u#0)&Bgt*J%H#tiPFplEE,S.tj:i+

8=?*l+ 8 i6Ppqkll J3 f=. EP E#%751 6

Ei-I-'DVLI _{I),JJiiU)I,JT^fS)I)1;}_'flfLtj_iL_-/IV)Tt. ]Ii.a''!.i.i5t.I.Wl(I

[?1tltL -/I_i._-i:J]'/J=L flo

I;-I:i7i-U7,I,

1. Satoshi Endo, Toshiyuki Matsunaga, Hiroaki Mamiya, Chisato Ohta, Midori Soda, Yukio Kitade, Kazuo

Tajima,

Hai-Tao Zhao, Ossama El-Kabbani _and Akira Hara, Kinetic _studies _{of AKRIB10,} human _aldose reductase-like protein: Endogenous substrates and inhibition by steroids, Arch. Biochem. Biophys., 487, 1-9

(2009) (IF; 3.046)

2. Satoshi Endo, Toshiyuki Matsunaga, Kazuo Kuwata, Hai-Tao Zhao, Ossama El-Kabbani, Yukio Kitade _and Akira Hara, Chromene-3-carboxamide derivatives discovered from _{virtual screening} as _potent inhibitors of

the tumour _{maker,AKRIB10,} Bioorg. Med. Chem., 18, 2485-2490 _(2010). (IF;2.822)

Satoshi Endo, Toshiyuki Matsunaga, Chisato Ohta, Midori Soda, Ayano Kanamori, Yukio Kitade, Satoshi Ohno, Kazuo

_Tajima,

Ossama El-Kabbani _{and Akira} Hara, Roles _of rat and human aldo-keto reductases in metabolism of famesol _{and geranylgeraniol,} Chem. Biol. Interact., _{in press} (DOI:

_{10.1016/j.cbi.2010.12.017)}

(IF; 2.457)

イソプレノイド代謝酵素を標的とする制癌薬開発の基礎研究

Title