archives of pharmacal research volume 25 issue 6 2002 [doi 10.1007_bf02976991] ki -jun hwang; kyung...

8/10/2019 Archives of Pharmacal Research Volume 25 Issue 6 2002 [Doi 10.1007_bf02976991] Ki -Jun Hwang; Kyung -Ho Par

1/5

Arch Pharm Res Vo l 25, No 6 , 781-785, 2002

~r~Oi~es of

barmatal ~e~eard

ht tp: / /apr.psk.or.kr

Nove l Benzoy lu rea Der iva t i ves as Po ten t i a l An t i tum or Agents ;

Syn thes is Ac t iv i ti es and S t ruc tu re -Ac t iv i ty Re la t ionsh ips

K i -J u n H w a n g K y u n g - H o P a r k 1 C h o n g - O c k L e e 2 a n d B e o m - T a e K i m

Depar tment o f Chem ist ry a nd R esearch Center o f B ioac t ive Ma ter ia ls , Co l lege o f Natura l S c ience , Chonbuk

Nat ional Univers ity , Dukj indong 66 4-14 Ch onju 561-756, Ko rea , lDup on t CR D, Experimental S tat ion, Wi lmington,

DE 198 80-0328, US A, and 2Screening Toxicology Research C enter, Korea Research Inst itute of Chem ical Tech-

no logy, P .O. Box 107, Yusun g, Tae jeon 305-606, Korea.

(Received Sep temb er 1, 2002 )

A ser ies o f pyrazo loxyphenyl benzoy l urea der ivat ives w as des igned a nd synthes ized for cy to-

tox ic eva luat ion as potent ia l ant itumor a gents . The synthet i c com pou nds we re eva luated for in

v i tro cy to tox ic i ty aga inst f ive hum an tum or ce l l l ines, inc lud ing A-549, SKO V-3, S K-ME L-2, X F-

498 and H CT-15 . Am o ng o t he rs , com pou nd 11 exh ib i ted 50 -10 0 t i m es g rea t e r an ti tum or

act ivi t ies than the commercial product , Cisplat in.

Key words : Benzoyl Urea, Ant icancer, Cisplat in, Cytotoxici ty

I N T R O D U C T I O N

A v a r i e t y o f p h e n y l b e n z o y l u r e a d e r i v a t i v e s a r e w e l l

k n o w n a s i n s e c t i c i d e s w h i c h i n h i b i t t h e c h i t i n s y n t h e s i s o f

i n s e c t s , a n d s o m e o f t h e m a r e c o m m e r c i a l l y a v a i l a b le .

W r i g h t et a l . , 1 9 8 7 ) . C h l o r f l u a z u r o n 1 ) i s a r e p r e s e n -

t a t iv e c o m m e r c i a l p r o d u c t w h i c h i s w i d e l y u s e d a s

i n s e c t i c i d e , W h i l e c h l o r f l u a z u r o n i t s e l f h a d n o a n t i t u m o r

a c t i v i t y , s i m i l a r b e n z o y l u r e a c o m p o u n d 2 w a s r e p o r t e d t o

p o s s e s s a n t it u m o r a c ti vi ty a g a i n s t p 3 8 8 l e u k e m i a O k a d a

et a l . ,

1 9 9 1 ) . T h e r e a r e a l s o s e v e r a l r e l e v a n t r e p o r t s

H o w b e r t et a l . , 1 9 9 0 , C h e r n e t a l . , 1 9 9 7 ) a b o u t t h e

d i a r y ls u l fo n y l u r e a c o m p o u n d s 3 w h i c h w e r e r e p o r te d t o

b e e f f e c ti v e a g a i n s t s o l i d t u m o r s .

A f t e r t h e r e p o r t b y O k a d a et a l . , v a r i o u s b e n z o y l u r e a

c o m p o u n d s w e r e f u r t h e r i n v e s t i g a t e d i n t h e s e a r c h o f

p o t e n t i a l a n t i c a n c e r d e r i v a t i v e s t o c o m e u p w i t h H O - 2 2 1

4 ) b y I sh i h a ra S h n g y o K a i s h a , L T D . K o n d o et al . , 1 9 9 3 ) .

A l t h o u g h H O - 2 2 1 w a s f o u n d t o a c t a s a s t r o n g i n h i b i t o r o f

D N A p o l y m e r a s e a l p h a a c t i v i t y a n d s e l e c t i e d a s a c a n -

d i d a t e c o m p o u n d t o b e d e v e l o p e d , i t s i n h e r e n t s o l u b i l i t y

p r o p e r t y h a s c a u s e d i t t o b e v i e w e d a s i n a p p r o p r i a t e f o r

u s e i n m e d i c i n a l d r u g . T h e c a r e f u l s c r u ti n it y o f H O - 2 2 1 b y

Correspondence to: Ki -Jun Hwang, Department o f Chemist ry ,

Col lege of Natura l Scienc e, Chonbu k Na t ional Universi ty, Dukj in-

dong 664-2, Chonju 561-756, Korea

E-mai l : k i jun@ moak.chonbuk.ac.kr

c o m p a r i n g w i t h t h e e a r l i e r l e a d c o m p o u n d 2 r e v e a l e d t h a t

t h e h e t e r o c y c l c l ic g r o u p s a t t a c h e d t o p h e n y l r i n g a f fe c t t h e

an t i cance r ac t i v i t y d ram at i ca l l y .

In c o n ti n u a ti o n o f o u r w o r k o n b e n z o y l u r e a c o m p o u n d s

H w a n g

et aL,

1 9 9 5 a n d 1 9 9 7 ) a n d in v ie w o f a b o v e m e n -

t io n e d l it e r a tu r e p r e c e d e n c e , i t w a s p l a n n e d t o u n d e r t a k e

t h e s y n t h e s is o f b e n z o y l u r e a c o m p o u n d s c o n t a in i n g n o v e l

p y r a z o l e h e t e r o c y l i c m o i e t y t o s e c u r e c o m p o u n d s o f e n -

h a n c e d a n t i t u m o r a c tiv it ie s . T h e p r e s e n t p a p e r d e s c r i b e s

s y n t h e s i s a n d b i o l o g i c a l e v a l u a t i o n o f t h e b e n z o y l u r e a

d e r i v a ti v e s c o n t a i n i n g n o v e l p y r a z o l e m o i e ti e s .

M T E R I L N D M E T H O D S

S y n t h e s i s

M e l t i n g p o i n t s w e r e r e c o r d e d o n e l e c t r o t h e r m a l m e l t i n g

p o i n t a p p a r a t u s a n d a r e u n c o r r e c t e d . M a s s s p e c t r a w e r e

r e c o r d e d o n a S h i m a d z u Q P - 1 0 0 0 s p e c t r o m e t e r 2 0 e V ) .

1 H - N M R a n d I ~ C - N M R d a t a w e r e o b t a i n e d f r o m J e o l 4 0 0

M H z s p e c t r o m e t e r a n d c h e m i c a l s h if ts 5 ) w e r e r e p o r te d

i n ppm i n re l a t ion t o t e t ram et hy l s i l ane 8 0 . 00 ) and C DCI 3

8 77 . 0 ) f o r 1H and ~ 3C NM R, respec t i ve l y . Th i n l aye r ch ro -

m a t o g r a p h y w e r e p e r f o rm e d o n p r e - c o a t e d s i li ca g e l 6 0 F -

2 5 4 l a y e r t h i c k n e s s 0 .2 m m , M e r c k) . T h e F l a s h C o l u m n

C h r o m a t o g r a p h y w a s p e r f o r m e d o n M e r c k s i l i c a g e l t y p e

6 0 2 3 0 - 4 0 0 m e s h ) . T h e o r g a n i c s o l v e n ts a n d c h e m i c a l s

w e r e o b t a i n e d f r o m A l d d c h . C o . a n d p u r i fi e d b y t h e a p p r o -

p r i a t e m e t h o d s b e f o r e u s e .

781


2/5


3/5

Novel Benzoy lurea Der ivat ives as Potent ia l Ant i tumor Agents 783

Table I . Che mical Structures and Physical Properties of Compound 8

No. W X Formula Y mp, yie ld,% ~H NMR(acetone-d6,

11a 4H CH3 C19H12CI2FsN3Os 3,5 -C l2 225-2 26

11b 4H t-bu tyl C22H~sCI2F3N~Os 3,5 -C l2 228-22 g

11c 4H CH3 Cl~H13ClF~NsO~ 3- C l 20 5-20 6

11d 4H t-bu tyl C22H19ClF3NsQ 3- C l 195-1 97

11e

4H P h e n y l C2sH~sF6NsO5 3-CF3 153-154

11f 4H CH~ C19H12CI2F~N~O~ 2, 5- C l2 20 8-20 9

1g 4H P h e n y l C24H~sCIF3N~O~ 3- C l 212-21 4

11h

5H CH~ C~gH12CI2F~N~O~ 3,5 -C I2 25 3-25 4

11i 5H CH3 C19H~CIF3N~Os 3- C l 20 1-20 2

70 3.9(s ,3H ), .8(s,lH), 7.8-8.3(m,6H), 10.5(s,1H), 10.7(s,lH)

65 1.8(s , H), 5.8(s,1H),7.8-8.3(m,7H), 10.5(s,1H), 10.7(s, 1H)

64 3.9(s ,3H ), .9(s,1H),7.4-8.3(m,7H), 10.4(s,1H), 10.6(s,1H)

66 1.8(s , H), 5.9(s,1H),7.4-8.3(m,7H),10.4(s,1H), 10.6(s,1H)

62 6.3 (s,1 H ), .4-8.3(m,12H), 10.4(s,1H), 10.7(s,1H)

70 3.9(s ,3H ), .1(s,1H),7.7-8.7(m,6H),10.6(s,1H),11.2(s,1H)

74 6.1 (s,1 H ), .5-8.3(m,12H), 10.4(s,1H), 10.6(s,1H)

73 3.8(s,3 H), .3(s,1H),7.8-8.3(m,7H),10.4(s,1H),10.6(s,1H)

71 3.9(s,3H),7.1-8 .3(m,8H ), 0.4(s,1H), 10.5(s,1H)

in 6 2 - 7 5 % y i e ld s .

2 - n i t r o - N - [ [ [ 3 , 5 - d i c h l o r o - 4 - ( 1 - m e t y l - 3 - t r i f l o r o m e t h y l -

5 - p y r a z o y l ) o x y p h e n y l ] a m i n o ] c a r b o n y l ] b e n z d m i d e ( 1 1 a ) .

Y i e ld 7 0 % ; m p 2 2 5 ; 1 H N M R ( a c e t o n -d 6 ) ~ 3 . 9 ( s , 3 H ) , 5 . 8

(s , 1H ) , 7 . 8 -8 . 3 (m , 6H ) , 10 . 5 (s , 1H ), 10 . 7 (s , l h ) ; M S : m /

z 5 1 8 ( M + ) .

T h e c h e m i c a l s t r u c t u r e s a n d t h e i r p h y s i c a l p r o p e r t ie s f o r

o t h e r c o m p o u n d s 1 1 a r e s u m m a r i z e d i n T a b l e I .

A n t i c a n c e r a c t iv i ty t e s t i n v i t r o )

A n t i c a n c e r a s s a y w a s p e r f o rm e d b y P h a r m a c e u t i c a l

S c r e e n i n g L a b o r a t o r y i n K o r e a R e s e a r c h I n s t i t u t e o f

C h e m i c a l T e c h n o l o g y u s i n g f i v e d i f f e r e n t h u m a n t u m o r

c e l l l i n e s , A - 5 4 9 ( h u m a n l u n g ) , S K - O V - 3 ( h u m a n o v a r i a n ) ,

S K - M E L - 2 ( h u m a n m e l a n o m a ) , H C T - 1 5 ( h u m a n c o l o n ) ,

X F - 4 9 8 ( h u m a n C N S ) w h i c h w e r e p u r c h a s e d f r o m t h e

Na t i ona l Cance r I ns t i t u t e (NC I ) i n U . S . A .

T h e c e l ls w e r e g r o w n a t 3 7 ~ in R P M I 1 6 4 0 m e d i u m

s u p p l e m e n t e d w i t h 1 0 % F B S a n d s e p a r a t e d u s i n g P B S

con t a i n i ng 0 . 25% t ryps i n and 3 m M EDTA. 5103-2104 ce l l s

w e r e a d d e d t o e a c h w e l l o f 9 6 w e l l p l a t e a n d i n c u b a t e d a t

3 7 ~ f o r 2 4 h . E a c h c o m p o u n d w a s d is s o l v e d in D M S O

a n d d i lu t e d w i t h th e a b o v e m e d i u m a t f iv e d i f fe r e n t c o n -

c e n t r a t i o n s w i t h t h e r a n g e o f 0 . 1 - 3 0 t ~ g / m L . T h e D M S O

c o n c e n t r a t i o n w a s s e t t o b e b e l o w 0 . 5 % a n d f i l t r a t e d

u s i n g 0 . 2 2 m g f il te r . A f te r r e m o v i n g t h e w e l l m e d i u m b y

a s p i r a ti o n , a 2 0 0 t.[ L p o r t io n o f t h e s o l u t io n w a s a d d e d t o

a b o v e w e l l p l a t e s w h i c h w e r e p l a c e d i n 5 % C O 2 i n c u b a t o r

f o r 4 8 h . T h e p r o t e in s t a i n a s s a y w a s p e r f o r m e d a c c o r d i n g

t o S R B m e t h o d ( S k e h a n

e t a l .

1990) .

R E S U L TS N D D I S C U S S I O N

C h e m i s t r y

T h e s y n t h e s i s o f t a r g e t c o m p o u n d s 1 1 w a s s t a r te d f r o m

t h e p r e p a r a t i o n o f p y r a z o y l o x y n i t r o b e n z e n e 7 b y c o n d e n s -

i n g c o m m e r c i a l ly a v a i la b l e c h l o r o n i t r o b e n z e n e 5 w i t h

p y r a z o l e s 6 u n d e r b a s i c c o n d i t io n a t 9 0 ~ 1 7 6 f o r 3 h

i n 6 5 - 7 5 % y ie l d ( S c h e m e 1 ). T h e r e q u ir e d 5 - h y d r o x y - 3 -

t r if iu o r o m e t h y l p y r a z o l e a n d 4 - h y d r o x y - 3 - t r if lu o r o m e t h y l -

p y r a z o l e w a s p r e p a r e d u t i l i z i n g t h e p r e v i o u s l y d e s c r i b e d

m e t h o d s b y B r u c e e t a l . 1 9 9 3 a n d I w a t a e t a l . 1 9 9 1 ,

respec t i ve l y .

T h e n i t r o c o m p o u n d s 7 w e r e t h e n r e d u c e d t o a n i l i n e

a n a l o g u e s b y tr e a ti n g th e m u n d e r n o r m a l h y d r o g e n a t io n

cond i t i on . Wi t h t he key i n t e r r r i ed i a t es 8 i n hand , t he f i na l

CI CI

Ch l o r f l uazu ron (1 )

, -R2

R I ~ ~ J H H

Fig. 1. Structuresof som e biologically active urea com pounds

C l 0 0 O

2

N O 2 0 0 > - -/ y O ~ l ~ Nx--~

I I I |

HO-221 (4)

B r


4/5

784 K. -J . Hwan g e t a l

W \ .C F 3

,,., I .. , II

4 . . . N

O 2N . - ' ' ~ . \ ~ N

O 2 N . ~ C I a b

Y Y X Y

5 7

W C F 3

R e a g e n t s a ) H o . ~ .~ [ , ( 6 ), D M F , K 2 C O 3 , 9 0 - 1 0 0 ~ 3 h

< N . N

X

b) Ran ey N i , H2 , 90~ 4h

Schem e 1. Synthesisof Ke y Intermediates Pyrazo yloxyanilines8

W

O \ C F 3

H 2 N ' ~ \ N

X

NO

a ~ [ ~ ~ NO2 C~..O

N H 2 ~ . . N ~

O O

9 10

Re age nts : a) oxa ly l ch lor ide, CICH2CH2C I , 100~ 2h

b)

H 2 N ~ W . .~. t~I /CF3

- -N~'~\'~L'O~h- N (8 ), rt, l h

Y

X

Scheme 2. Synthesisof Benzoyl Ure a Derivatives 11

N O 2

H H 2 W

N -.rr. N 1 -.~ ~3 \ /C F3

II II I

4 \-~Trj

O O U \ / .L4 I I

11 X

p r o d u c t s 1 1 w e r e p r e p a r e d w i t h n o i n c i d e n c e b y r e a c t i n g

8 w i t h n i t r o b e n z o y l i s o c y a n a t e 9 w h i c h w a s f o r m e d i n s i t u

f r o m n i t r o b e n z a m i d e a n d o x a l y l c h l o r i d e ( S c h e m e 2 ) .

A n t i c a n c e r a s s a y

T h e a n t i t u m o r a c t i v it ie s o f th e c o m p o u n d 1 1 to w a r d f iv e

d i f f e r e n t h u m a n t u m o r c e l l s w a s d e t e r m i n e d f r o m t h e T z

( n u m b e r o f c e ll s b e f o r e t h e a d d i t io n o f th e t e s t c o m p o u n d s ) ,

C ( n u m b e r o f c e l ls a f t e r 4 8 h i n c u b a t i o n w i t h o u t t e s t

c o m p o u n d s ) , a n d T ( n u m b e r o f c e l l s a f t e r 4 8 h i n c u b a t i o n

w i t h t e s t c o m p o u n d s ) b y t h e f o ll o w i n g e q u a t i o n ( 1 ) o r ( 2) .

~ - ( T - T z ) ~

I f T z > T , , ( C _ - - ~ z ) j X 1 0 0 ( 1 )

If T z < T , [ ~ ] x l 0 0 ( 2 )

T h e v a l u e t h u s o b t a i n e d w e r e a p p l i e d t o L o t u s p r o g r a m

u s i n g d a t a r e g r e s s i o n t o f iv e E D s o o f e a c h c o m p o u n d . T h e

r e s u l t o f t h e i n v i tr o a n t i c a n c e r d a t a a r e c o m p a r e d t o c o m -

m e r c i a l p r o d u c t c i s p l a t i n ( S i g m a T a u C o , I t a l y ) a n d s u m -

m adzed i n Tab l e I I .

T h e r e s u l t s i n d ic a t e e x c e l l e n t a n t i t u m e r a c t i v it ie s t o w a r d

t he f i ve d i f f e ren t ce l l l i nes , A -549 ( l ung ca rc i nom a) , SK-

O V - 3 ( o v a r y m a l i g n a n t a s c i t e s ) , S K - M E L - 2 ( m a l i g n a n t

m e l a n o m a ) , X F - 4 9 8 ( C N S t u m o r ) a n d H C T - 1 5 ( c o l o n a d -

e n o c a r c i n o m a ) . A d d i t i o n a l ly c o m p o u n d 1 1 g e x h ib i ts 5 0 -

1 0 0 t i m e s m o r e p o t e n t t h a n c o m m e r c i a l p r o d u c t c i s p l a t i n

Tab le I1. Antitumu or Activites of targ et com pou nd 11

Compound EDs0 l~g/mL)

N~ A -549 SK-O V-3 SK-M EL-2 XF 498 HCT 15

Cisplatinb 0.7 75 0.460 0,460 0.638 1.100

11a 0.281 0.332 0.242 0.494 0.202

11b 4.7 95 24 .31 4 7.245 7.225 7.489

11c 0.256 0.281 0.139 0.129 0.022

11d 2.611 4.284 2.386 4.431 2.701

11e 1.137 1.597 0.640 1,763 0.880

11f 0.614 1.212 0.429 2.059 0.383

11g 0.916 3.301 1.059 2.282 0.951

11h 2.993 3.041 1.960 3.644 1.929

11i 0.011 0.023 0.022 0.029 0,013

refer to Table I.

b taly, Sigm a Tau

t o w a r d t h e f i v e h u m a n c a n c e r c e l l l i n e s . N o w i t c a n b e

c l e a r l y s t a t e d t h a t b e n z o y l u r e a d e r i v a t i v e s c o n t a i n i n g

h e t e r o c y c l i c p y r a z o l g r o u p p o s s e s s e x c e l l e n t a n t i t u m o r

ac t i v i ti es i n v it ro , a nd f u r t he r s t udy i nc l ud i ng i n v i vo ex pe r i -

m e n t s a n d m o l e c u l a r m o d i f i c a t i o n s i s w a r r a n t e d .

R E F E R E N E S

Chern, J . W . , Leu , Y. , W ang , S. , Jou, R. , Lee , C.F. , Tsou, P.C. ,


5/5

Nove l Benzoy lu rea De r iva t ives as Poten t ia l An t i tum or Age nts 785

Hsu, S .C. , L iaw, Y .C. and L in , H.M. , Syn thes is and cy to tox ic

eva lua t ion o f subst i tu ted su l fony I -N-hy droxy gua n id ine der iva-

t ives as po ten t ia l an t i tumor agents . J . M e d . C h e m . 4 0 , 2 2 7 6 -

2286 1997) .

Gaede, B. and McDermott , L. L. , Novel Perf luoroalkyl-subst i tut-

ed pyrazo les .

J . H e t e r o cyc l i c C h e m .

30, 49-54 1993) .

Howbert , J. J. , Grossman, S. Crowel, T.A., Rieder, B. J. , Harper,

R. W. , K ram er , K . E . Tao , E . V . , A ik ins , J . , Poore , G. A . ,

R inze l , S . M. , Gr indey, G. B . , Shaw, W. N. , and Todd, G. C. ,

Nove l age nts e f fec t ive aga ins t so l id tum or : The ary lsu l f-

ony lu reas, syn thes is , ac t iv i t ies , and ana lys is o f QSAR. J .

M e d . C h e m .

3 , 2 3 9 3 - 2 4 0 7 1 9 9 0 ).

Hwang, K . J . and Park , K . H. pyrazo le der iva t ives , U S p a t e n t

5 4 3 0 1 5 8 1 9 9 5 ) .

Hwang, K . J . and Park , K . H. pyrazo le con ta in ing benzoy lu rea

der iva t ives , compos i t ion and use , E P 0557481 1997) .

Iwata, S. , Namekata, J. , Tanaka, K. and Mitsuhashi, K. , Synthesis

of 4-Hydroxy-3- tr i f luoromethypyrazoles.

J. Heterocyc l i c Chem.

28, 1971-1976 1991) .

Kondo, N., Iwao, T. , Masuda, H, Yamanouchi, K. , Ishihara, Y. ,

Yamad a, N. , Haga, T ., Og aw a, Y . and Yokoyam a, K . ,

Improved o ra l absorp tion o f a poor ly water-so lub le d rug , HO -

221. C h e m . Ph a r m . Bu l l. 41 4) , 737-740 1993) .

Okada, H. , Koyanag i , T . , Yamada, N. and Haga, T . , Syn thes is

and an t i tum or ac t iv ies o f nove l benzoy lpheny lu rea , der iva-

t ives . C h e m . Ph a r m . Bu l l. 39 9) , 2308-2315 1991) .

Skehan, P . , S to reng, R . , Scud ie ro , D. , Monks, A . , McM aho n, J .

B., V ist ica, D. T. , Warren, J. T. , Bokesch, H., Kenn, S.and

Boyd, M. R . , J . N a t l . C a n ce r l n s t . 82, 1107 1990) .

W r igh t , J . E . and Ret rakaran , A . eds. ) , C h i t in and Be nzoy l -

pheny l u reas, Dr . W . Junk pub l ishers , Dordrech t , 1987) .

archives of pharmacal research volume 25 issue 6 2002 [doi 10.1007_bf02976991] ki -jun hwang; kyung...

Documents