Synthetic Oligosaccharides - American Chemical Society

Chapter 8

Synthetic Acceptors for α-L-Fucosyltransferases

Downloaded by UNIV MASSACHUSETTS AMHERST on August 19, 2012 | http://pubs.acs.org Publication Date: May 5, 1994 | doi: 10.1021/bk-1994-0560.ch008

Khushi L. Matta, Conrad F. Piskorz, Gurijala V. Reddy, Ε. V. Chandrasekaran, and Rakesh K. Jain Department of Gynecologic Oncology, Roswell Park Cancer Institute, Buffalo, NY 14263

An elegant synthesis of Galβ1->3 (α-L-Fuc1->4) GlcNAcβ -O-allyl and 3-O-SO Na .GalB1->3 (α-L-Fucl->4) GlcNAcβ-O - a l l y l have been accomplished through use of a key glycosyl donor, namely, methyl 2,3,4-tri-O-(4methoxybenzyl)-1-thio-β-L-fucopyranoside. These compounds can be employed as unique oligosaccharide ligands for human selectin molecules. -

+

3

+

The s y n t h e s i s o f 3 - 0 - S O ~ N a . G a l B l - > 3 / 4 G l c N A c B - 0 R a s an a c c e p t o r f o r α - L - f u c o s y l t r a n s f e r a s e i s a l s o d e s c r i b e d . The e x p r e s s i o n o f L - f u c o s e c o n t a i n i n g c a r b o h y d r a t e s t r u c t u r e s i n a m a j o r i t y o f human c a n c e r s ( 2 - 5 ) p r o v o k e d a n enormous i n t e r e s t i n t h e c h a r a c t e r i z a t i o n o f a - L fucosyltransferase a c t i v i t i e s present i n various t i s s u e s and c e l l l i n e s . Such i n v e s t i g a t i o n s have even l e d t o t h e d i s c o v e r y o f new t y p e s o f α - L - f u c o s y l t r a n s f e r a s e s b a s e d on t h e i r r e q u i r e m e n t o f s p e c i f i c s t r u c t u r e s in the acceptor molecules for optimal a c t i v i t y (2,6-27). L i k e w i s e , t h e b i o s y n t h e t i c pathways o f t h e s e fucosyl linked glycoconjugates have become a target of i n v e s t i g a t i o n s i n many l a b o r a t o r i e s . A n a b e r r a t i o n i n a L - f u c o s y l a t i o n as w e l l as α ( 2 , 3 ) - s i a l y l a t i o n a p p e a r s t o be r e s p o n s i b l e f o r t h e s y n t h e s i s o f s i a l y l L e a n d s i a l y l L e type structures. Thus, t h e r e has been a surge o f i n t e r e s t i n the study of g l y c o s y l t r a n s f e r a s e s i n normal a n d t u m o r t i s s u e s and v a r i o u s c e l l l i n e s . Compared t o t h e s i a l y l a t e d and f u c o s y l a t e d g l y c o c o n j u g a t e s r e p o r t s o n c a n c e r - a s s o c i a t e d sulfoglycoconjugates are scanty (2824). Nevertheless, these anionic glycoconjugates, e s p e c i a l l y t h e s u l f a t e d m u c i n t y p e s t r u c t u r e s (25-29) 3

x

a

0097-6156/94/0560-0120$08.00/0 © 1994 American Chemical Society

In Synthetic Oligosaccharides; Ková, P.; ACS Symposium Series; American Chemical Society: Washington, DC, 1994.

8. MATTA ET AL.

Synthetic Acceptors for a-L-Fucosyltransferases

121


h a v e a t t r a c t e d much r e c e n t a t t e n t i o n . Interest i n the s u l f o g l y c o c o n j u g a t e s h a s f u r t h e r stemmed f r o m t h e f a c t t h a t t h e s e compounds c o n t a i n i n g s u l f a t e , f u c o s e a n d e v e n s i a l i c a c i d a r e b e i n g r e p o r t e d t o be l i k e l y l i g a n d s f o r s e l e c t i n s , t h e f a m i l y o f c e l l a d h e s i o n m o l e c u l e s (29-35) . An e x a m i n a t i o n o f t h e s t r u c t u r e o f s u l f a t e d a n d s i a l y l a t e d oligosaccharides, e s p e c i a l l y those occurring i n g l y c o l i p i d s ( 3 6 ) , r e v e a l s t h a t s u l f a t e g r o u p s i n most of t h e s e g l y c o c o n j u g a t e s a r e g e n e r a l l y l o c a t e d where sialic acid occurs i n the corresponding s i a l y l a t e d g l y c o l i p i d s a n d g l y c o p r o t e i n s . We h a v e o b s e r v e d t h a t t h e s p e c i f i c i t y o f α - L - f u c o s y l t r a n s f e r a s e (27) i s s i m i l a r f o r b o t h s u l f a t e d and c o r r e s p o n d i n g s i a l y l a t e d a c c e p t o r s . B a s e d u p o n t h e s e o b s e r v a t i o n s , we h a v e s y n t h e s i z e d a l l y l 0-(3-0-sulfo-B-D-galactopyranosyl)-(l->3)-2-acetamido-2d e o x y - B - D - g l u c o p y r a n o s i d e s o d i u m s a l t (13) , a l l y l 0 - ( B - D galactopyranosyl) - ( l - » 3 ) -[O-(a-L-fucopyranosyl)-(1-+4) ]-2a c e t a m i d o - 2 - d e o x y - e - D - g l u c o p y r a n o s i d e (17) , a l l y l 0 - ( 3 - 0 s u l f ο - β - D - g a l a c t o p y r a n o s y l ) - ( l->3 ) - [ 0 - ( α - L - f u c o p y r a n o s y l ) (1-+4)]-2-acetamido-2-deoxy-B-D-glucopyranoside sodium salt (24), b e n z y l 0 - ( 3 - 0 - s u l f o - B - D - g a l a c t o p y r a n o s y l ) ( 1 - + 3 ) - 2 - a c e t a m i d o - 2 - d e o x y - B - D - g l u c o p y r a n o s i d e sodium s a l t (25) , and b e n z y l 0-(3-0-sulfο-β-D-galactopyranosyl)( l - * 4 ) - 2 - a c e t a m i d o - 2 - d e o x y - B - D - g l u c o p y r a n o s i d e sodium s a l t

(26) . Compounds 13, 25 and 26 c a n be u s e d a s s u b s t r a t e s f o r α - L - f u c o s y l t r a n s f e r a s e a n d 17 a n d 24 a s l i g a n d s f o r s e l e c t i n molecules. The k e y i n t e r m e d i a t e 12 was p r e p a r e d i n s i x s t e p s from the known disaccharide allyl 0-(B-Dgalactopyranosyl)-(l->3)-2-acetamido-2-deoxy-B-Dg l u c o p y r a n o s i d e (37) 8, w h i c h u p o n t r e a t m e n t w i t h t e r t b u t y l c h l o r o d i p h e n y l s i l a n e (38), f o l l o w e d by a c e t a l a t i o n w i t h 2,2-dimethoxypropane-acetone i n the presence o f 4toluenesulfonic acid, then the removal of the tertbutyldiphenylsilyl group w i t h IM t e t r a b u t y l a m m o n i u m f l u o r i d e i n oxolane (38), p r o v i d e d 3 » , 4 - 0 - i s o p r o p y l i d e n e compound. Acetylation with p y r i d i n e - a c e t i c anhydride, f o l l o w e d by c l e a v a g e o f t h e 3 , 4 - 0 - i s o p r o p y l i d e n e g r o u p with c h l o r o f o r m - t r i f l u o r o a c e t i c acid-water furnished the d i o l 11 i n 71% y i e l d . The d i o l 11 was c o n v e r t e d i n t o i t s 3 , 4 - ( e t h y l o r t h o a c e t a t e ) , w h i c h was h y d r o l y s e d t o g i v e a key intermediate, allyl 0-(2,4,6-tri-0-acetyl-B-Dg a l a c t o p y r a n o s y l ) - (1-K3) - 2 - a c e t a m i d o - 4 , 6 - d i - 0 - a c e t y l - 2 d e o x y - B - D - g l u c o p y r a n o s i d e 12 i n 75% y i e l d . The ^ H - N . M . R . s p e c t r u m o f 12 e x h i b i t e d a low f i e l d c h e m i c a l s h i f t a t £ 5 . 2 8 ( d , J = 3 . 7 H z ) , c o n f i r m i n g t h a t d i o l 11 h a d b e e n a c e t y l a t e d a t Ο ' - 4 t o g i v e 12. S u l f a t i o n o f 12 w i t h sulfur t r i o x i d e - p y r i d i n e complex followed by de-0acetylation with methanolic sodium methoxide gave compound 13. A s i m i l a r r e a c t i o n s e q u e n c e was e m p l o y e d 1

1

1


SYNTHETIC OLIGOSACCHARIDES

122

for the synthesis of benzyl 0-(2,4,6-tri-0-acetyl-B-Dg a l a c t o p y r a n o s y l ) - (l->3) - 2 - a c e t a m i d o - 4 , 6-di-0-acetyl-2deoxy-B-D-glucopyranoside, a key i n t e r m e d i a t e f o r the compound 25, and benzyl 0-(2,4,6-tri-0-acetyl-B-Dg a l a c t o p y r a n o s y l ) - (l->4) - 2 - a c e t a m i d o - 3 , 6-di-0-acetyl-2deoxy-B-D-glucopyranoside, a key i n t e r m e d i a t e for the compound 26, a s t h a t d e s c r i b e d f o r t h e p r e p a r a t i o n o f 13 f r o m 8 (See Scheme 1 ) . T h e s t r u c t u r e o f 13, 25 a n d 26 was c o n f i r m e d b y C - N . M . R . spectroscopy (see T a b l e I) and FABMS (see E x p e r i m e n t a l S e c t i o n ) . Downloaded by UNIV MASSACHUSETTS AMHERST on August 19, 2012 | http://pubs.acs.org Publication Date: May 5, 1994 | doi: 10.1021/bk-1994-0560.ch008

1 3

Methyl 2 , 3 , 4 - t r i - 0 - b e n z y l - l - t h i o - B - L - f u c o p y r a n o s i d e , 2 , 3 , 4 - t r i - 0 - b e n z y l - B - L - f u c o p y r a n o s y l f l u o r i d e , and 2 , 3 , 4 t r i-0-benzyl-a, B-L-fucopyranosyl t r ichloroacetamidate (39-43) c a n n o t be u t i l i z e d f o r t h e s y n t h e s i s o f compound 17 a n d 24 s i n c e t h e y r e q u i r e h y d r o g e n o l y s i s f o r r e m o v a l of the benzyl p r o t e c t i n g groups. Methyl 2 , 3 , 4 - t r i - 0 - ( 4 methoxybenzyl)-l-thio-B-L-fucopyranoside 2 provides a more e f f i c i e n t r o u t e f o r α - L - f u c o s y l a t i o n b e c a u s e r e m o v a l of t h e p r o t e c t i n g groups does not r e q u i r e h y d r o g e n o l y s i s . Compound 2 was o b t a i n e d t h r o u g h a l k y l a t i o n o f m e t h y l - 1 t h i o - B - L - f u c o p y r a n o s i d e 1 with 4-methoxybenzyl c h l o r i d e p o t a s s i u m h y d r o x i d e - 1 8 - c r o w n - 6 (44) i n 68% y i e l d . Allyl 0 - ( 2 , 3 , 4 , 6 - t e t r a - 0 - a c e t y l - B - D - g a l a c t o p y r a n o s y l ) - (1-+3) - 2 acetamido-6-0-benzyl-2-deoxy-B-D-glucopyranoside 7 was o b t a i n e d by t h e r e d u c t i n g r i n g o p e n i n g (45) o f t h e 4 , 6 - 0 b e n z y l i d e n e a c e t a l g r o u p o f 6 i n a c i d i c medium, i n t h e presence of sodium cyanoborohydride in 65% yield. G l y c o s y l a t i o n under c u p r i c bromide and tetrabutylammonium b r o m i d e (46) c o n d i t i o n o f compound 7 w i t h 2 g a v e 14 i n 68% y i e l d after silica gel column chromatography. Removal o f 6 - 0 - b e n z y l g r o u p i n 14 w i t h 1% ( v / v ) of trimethylsilyl trifluoromethane sulfonate in acetic a n h y d r i d e was n o t s u c c e s s f u l t o g i v e compound 16. De-0acetylation of compound 14 w i t h methanolic sodium methoxide followed by t h e treatment with 1% (v/v) trimethylsilyl trifluoromethane sulfonate in acetic a n h y d r i d e (47) g a v e f u l l y a c e t y l a t e d t r i s a c c h a r i d e 16 i n 61% y i e l d . De-O-acetylation with methanolic sodium m e t h o x i d e p r o v i d e d compound 17 i n 89% y i e l d ; t h e C N.M.R. spectrum was in accord with the structure assigned. T r e a t m e n t o f compound 4 w i t h p i v a l o y l c h l o r i d e i n p y r i d i n e p r o v i d e d t h e 6 - 0 - p i v a l o y l compound 5 i n 90% y i e l d ; i t s H - N . M . R . s p e c t r u m was i n a g r e e m e n t w i t h t h e s t r u c t u r e proposed. A s i m i l a r g l y c o s y l a t i o n of 5 with 2 a f f o r d e d t r i s a c c h a r i d e d e r i v a t i v e 18 i n 53% y i e l d , the ^-N.M.R. s p e c t r u m o f w h i c h showed a d o u b l e t a t 5 5 . 3 8 w i t h s p a c i n g of 3.1 Hz, c o n f i r m i n g the α - D - c o n f i g u r a t i o n of t h e new i n t e r g l y c o s i d i c b o n d . Removal o f t h e 4m e t h o x y b e n z y l g r o u p f r o m compound 18 w i t h e e r i e ammonium nitrate (48) in acetonitrile and followed by the t r e a t m e n t w i t h t r i e t h y l o r t h o a c e t a t e and d e - 0 - a c e t y l a t i o n 1

1


3

8.

MATTA ET AL.


with methanolic sodium methoxide afforded 3",4"e t h y l o r t h o a c e t a t e d e r i v a t i v e w h i c h on h y d r o l y s i s w i t h 80% a q u e o u s a c e t i c a c i d g a v e t h e 4 " - 0 - a c e t y l d e r i v a t i v e (20) i n 46% y i e l d ; t h e ^ - N . M . R . s p e c t r u m o f w h i c h showed a d o u b l e t a t 5 5 . 1 5 , c o n f i r m i n g t h a t 20 h a d b e e n a c e t y l a t e d a t 0"-4. I s o p r o p y l i d e n a t i o n o f compound 20 u n d e r C a t e l a n i et a l . (49) p r o c e d u r e a f f o r d e d 3 ,4 -0-isopropylidene d e r i v a t i v e 21. D e - O - a c e t y l a t i o n o f compound 21 f o l l o w e d by acetylation with pyridine-acetic anhydride and h y d r o l y s i s w i t h 60% aqueous a c e t i c a c i d p r o v i d e d d i o l 22 i n 55% y i e l d . I t was c o n v e r t e d t o k e y i n t e r m e d i a t e 23 by the orthoester procedure which on sulfation with s u l f u r t r i o x i d e - p y r i d i n e complex and d e - 0 - a c e t y l a t i o n w i t h m e t h a n o l i c s o d i u m m e t h o x i d e g a v e compound 24 a s s o d i u m s a l t a f t e r p a s s i n g t h r o u g h IR 120 (Na ) resin. The s t r u c t u r e o f 24 was c o n f i r m e d by C - N . M . R . and FAB mass s p e c t r o s c o p y (See Scheme 2; T a b l e I ) . 1


123

1

+

1 3

1 3

1 3

C - N . M . R . a s s i g n m e n t s - The a s s i g n m e n t o f t h e C-N.M.R. r e s o n a n c e s f o r t h e t h r e e s u l f a t e d d i s a c c h a r i d e s 13, 25 and 26 were made by c o m p a r i n g t h e i r s p e c t r a w i t h t h a t o f compound benzyl 0- ( B - D - g a l a c t o p y r a n o s y l ) - (l-*3) -2acetamido-2-deoxy-B-D-glucopyranoside reported i n Table I. In the C-N.M.R. spectra of 13, 25 and 26 a d o w n f i e l d s h i f t o f 7 . 5 5 - 7 . 7 8 p . p . m . was o b s e r v e d i n t h e r e s o n a n c e f o r C - 3 , a l o n g w i t h an u p f i e l d s h i f t o f 1 . 6 2 1.74 p . p . m . f o r C - 4 , i n c o m p a r i s o n t o t h e s p e c t r u m o f u n s u l f a t e d d i s a c c h a r i d e , c o n f i r m i n g 0-3 a s t h e s i t e o f s u l f a t i o n i n t h e s e compounds. I n t h e s p e c t r u m o f 24 a d o w n f i e l d s h i f t o f 7 . 9 p . p . m . was o b s e r v e d f o r t h e C - 3 resonance i n comparison to the spectrum of the p a r e n t compound 17 e v i d e n c i n g 0-3 as t h e s i t e o f s u l f a t i o n . 1 3

1

1

1

1

Synthesis of allyl 0-(3-0-sulfo-B-Dg a l a c t o p y r a n o s y l ) - (l-*3)-2-acetamido-2-deoxy-B-Dg l u c o p y r a n o s i d e sodium s a l t ( 1 3 ) . G l y c o s y l a t i o n o f a l l y l 2-acetamido-2-deoxy-4,6-0-(4-methoxybenzylidene)-B-Dglucopyranoside with 2,3,4,6-tetra-O-acetyl-a-Dg a l a c t o p y r a n o s y l bromide, i n the presence of mercury c y a n i d e , g a v e compound 3 w h i c h a f t e r r e m o v a l o f a c e t a l and acetyl groups afforded known allyl 0-(B-Dg a l a c t o p y r a n o s y l ) - ( l->3 ) - 2 - a c e t a m i d o - 2 - d e o x y - B - D g l u c o p y r a n o s i d e (8) (3 7). Compound 8 was t r e a t e d w i t h tert-butylchlorodiphenylsilane (38) in N,Ndimethylformamide to give allyl 0-(6-0-tertbutyldiphenylsilyl-B-D-galactopyranosyl)-(1-+3)-2acetamido-6-0-tert-butyldiphenylsilyl-2-deoxy-B-Dglucopyranoside (9) i n 68% y i e l d ; [a] - 1 3 ° (c 0.8, CHC1 ) : ^ - N . M . R . (CDCI3) : S 7 . 6 9 - 7 . 2 5 (m, 20 H , a r o m . ) , 6.24 ( d , J = 7 . 6 H z , 1 H , N H ) , 5 . 9 5 - 5 . 8 5 (m, 1 H , = C H ) , 5 . 1 6 ( d , J = 10.5 H z , 1 H , H - l ) , 4.78 ( d , J = 8 . 4 H z , 1 H , H - l ) , 2 . 1 6 ( s , 3 H , N A c ) , 1.97 and 0.99 ( e a c h s , 18 H , 2 χ CMe ) . D

3

1

3



C-N.M.R. data

a,b

z

C-1

C-2

Chemical Shifts

(Proposed Assignment)

C-3

C-4

a

3

3

3

3

4

Gal-B-(l-O) 102.41 70.45 71.44 67.45 GlcNAc-B-OBn 98.65 53.53 81.35 67.68 Compound 13 3-0-SO NaGal-B-(l-3) 102.25 69.45 79.15 65.71 GlcNAc-e-O-Allyl 98.81 53.35 81.86 67.80 Compound 17 Gal-6-(l-3) 101.82 69.49 71.32 66.77 Fuc-a-(l->4) 97.03 67.33 68.12 70.92 GlcNAc-e-OAllyl 98.93 54.67 75.09 74.44 Compound 24 3-0-SO NaGal-B-(l-3) 101.53 69.57 79.22 66.83 Fuc-a-(l-4) 97.00 67.62 68.15 70.94 GlcNAc-B-OAllyl 98.97 54.68 75.18 74.44 Compound 25 3-0-SO NaGal-B-(l->3) 102.26 70.82 79.22 65.83 GlcNAc-B-OBn 98.80 53.34 81.77 67.92 Compound 26 3-0-SO NaGal-fi-(l-4) 101.44 70.46 78.99 65.80 GlcNAc-B-OBn 98.84 54.04 71.31 77.39 For solutions in DO with Me Si as the external satisfactory elemental analysis.

Residue

13

TABLE I

59.88 59.75 60.00 14.33 58.75 60.46 14.33 58.77 59.98 59.88

73.84 74.36 73.80 65.81 71.43 73.48 65.82 71.34 73.93 74.50

21.31

21.31

21.24

21.21

21.15

NAc

132.38, 117.23

132.34, 117.18

132.33, 117.24

O-Allyl

b

73.76 59.88 73.88 59.07 21.11 standard; A l l products described gave

59.96 59.75

C-6

74.21 74.36

C-5



6

4

4

NHAc 4

5

Ri = H; R2=Bn

6

6 R\R2=C H CH

3

5 R< = H;R2=COCMe

4 R\R2=H

e

2

b

,

2

h

4

NHAc 4

2

4

3

4

2

r

4

3

3

V l 3 R'=R?=R = H;R3=S0 Na

4

R' = R2=R =Ac;R3=:H

4

5 11 R* = R*= Ac; R*=R =H

2

f 10 R' = H; R* = Bu'Ph SI; R , R = CMe

3

; 9 R' = R =R = H; R = BtfPh^l

3

^ 8 Ri = R2=R3=R =H

^12

fl

,

d

2

2

OCH CH=CH 2

Scheme 1

Reagents: a, KOH - MeOBnCI -18 - Crown - 6; b, 60% Aq AcOH; c, pyridine - CMe COCI; d, Bu«Ph SiCI - Imidazole - DMF β, DMP - Acetone - pTSA; f, IM Bu NF, pyridine - Acetic anhydride, g, Triethylorthoacetate/80% Aq AcOH; h, S0 -pyridine complex - DMF, MeOH - MeONa I. NaCNBH Ethereal-HCI

OCH CH=CH

3 R% R*=MeO*C H CH

OAc

2

CH C H «OMe< >

SMe


In Synthetic Oligosaccharides; Ková, P.; ACS Symposium Series; American Chemical Society: Washington, DC, 1994. 2

6

4

2

2

t

6

4

3

h, S0 -pyridine complex - DMF

2

2

2

4

=

R5 = Re H

3

3

4

5

Re- Ac 3

20 R* = COCM*,; R*= R = R = R = H;

R3, R = CMe

4

2 4

3

= S0 Na

=

=

24 Rt= R 2 = R 4 R 5 R 6 - H ; R 3

23 R ' = R? = R< = R* = R« = Ac; R3 = H

22 Ri = R2 = R5= Re = Ac; R3 = R = H

< 2

e

19 R* = COCMe ; R = R3= R« = Ac;

2

R5 - Re - CH C H · OMe(4)

2

j*21 R' = COCMe ; R 2 = R 5 = H ; R « = Ac;

e(

3

2

OCH CH=CH

18 R' = COCMe ; R?=R3= R3) - 2 - a c e t a m i d o - 6 - 0 - b e n z y l - 2 - d e o x y β - D - g l u c o p y r a n o s i d e 7 i n 65% y i e l d , [ a ] + 9 ° (c 1 . 2 , CHC1 ); ^-H-N.M.R. (CDC1 ) : 7 . 4 2 - 7 . 3 0 (m, 5 H , a r o m . ) , 5 . 9 0 - 5 . 6 9 (m, 1 H , =CH), 2 . 1 5 - 1 . 9 8 ( c l u s t e r o f s , 15 H , D

3

3


128


1 3

4 x OAc a n d N A c ) ; C - N . M . R . (CDC1 ) : 101.36 (C-1'), 98.09 ( C - 1 ) , 83.36 ( C - 3 ) , 66.96 ( C - 6 ) , 61.54 (C-6 ). Methyl 2,3,4-tri-0-(4-methoxybenzyl)-l-thio-B-Lfucopyranoside (2) was p r e p a r e d b y t h e a l k y l a t i o n o f 1 w i t h 4 - m e t h o x y b e n z y l c h l o r i d e - K O H - 1 8 - C r o w n - 6 (44) i n 68% y i e l d ; [ a ] - 8 ° ( ç 1 . 0 , CHC1 ) : ^ - N . M . R . (CDC1 ) : 6 7 . 3 0 ( d , 6 H , a r o m . ) , 6.80 ( d , 6 H , a r o m . ) , 3.80 ( s , 9 H , 3 χ OMe), 2 . 1 7 ( s , 3 H , S M e ) , 1.13 ( d , 3 H , C M e ) . 3

f


D

3

3

Reaction of 7 with 2 i n the presence of c u p r i c b r o m i d e - t e t r a b u t y l a m m o n i u m b r o m i d e (46) g a v e i n 68% y i e l d t h e f u l l y p r o t e c t e d t r i s a c c h a r i d e d e r i v a t i v e 14; [ a ] 5 8 ° (c 1 . 4 , CHC1 ) ; ^ - N . M . R . (CDC1 ) : δ 7 . 3 3 - 7 . 2 0 (m, 11 H , a r o m . ) , 6 . 8 9 - 6 . 7 8 (m, 6 H , a r o m . ) , 6 . 5 0 ( d , J = 9 . 3 H z , 1 H , N H ) , 5 . 8 7 - 5 . 8 3 (m, 1 H , = C H ) , 5 . 3 7 ( d , J = 3 . 6 Hz, 1 H , H - l " ) , 4.81 (d, J = 7.9 H z , 1 H , H - l ) , 3 . 8 1 , 3 . 7 9 a n d 3 . 7 6 ( e a c h s , 9 H , 3 χ OMe), 2 . 0 4 - 1 . 6 7 ( c l u s t e r o f s , 15 H , 4 χ OAc a n d N A c ) , 1.14 ( d , J = 6 . 5 H z , 3 H , CMe) . D

3

3

1

De-O-acetylation o f compound 14 w i t h methanolic s o d i u m m e t h o x i d e g a v e 15 i n 65% y i e l d ; [ a ] - 6 1 ° (c 1 . 2 , CHCI3) ; ^ - N . M . R . (CDCI3) : δ 7 . 3 8 - 7 . 2 1 (m, 11 H . a r o m . ) , 6 . 9 2 - 6 . 7 9 (m, 6 H , a r o m . ) , 3 . 8 0 , 3 . 7 9 a n d 3 . 7 6 ( e a c h s , 9 H , 3 χ OMe), 1.69 ( s , 3 H , N A c ) , 1.09 ( d , J = 5 . 9 H z , 3 H , CMe). D

Treatment of 15 i n a c e t i c anhydride-methylene c h l o r i d e ( 2 : 1 v / v ) c o n t a i n i n g 1% ( v / v ) o f t r i m e t h y l s i l y l t r i f l u o r o m e t h a n e s u l f o n a t e a t 0°C gave a l l y l 0 - ( 2 , 3 , 4 , 6 t e t r a - O - a c e t y l - B - D - g a l a c t o p y r a n o s y l ) - ( 1-+3) - 0 - [ 0 - ( 2 , 3 , 4 tri-O-acetyl-a-L-fucopyranosyl)-(l-»4)]-2-acetamido-6-0a c e t y l - 2 - d e o x y - B - D - g l u c o p y r a n o s i d e 16 i n 61% y i e l d ; [ a ] - 9 2 ° (c 1 . 2 , CHCI3) ; ^ H - N . M . R . (CDCI3) : δ 5 . 6 7 ( d , J = 8.3 H z , 1 H , NH), 5.40 (d, J = 3.4 H z , 1 H , H - l " ) , 4.87 ( d , J = 8 . 3 H z , 1 H , H - l ' ) , 2 . 1 6 - 1 . 9 6 ( c l u s t e r o f s , 27 H , 8 χ OAc a n d N A c ) , 1.24 ( d , J = 6 . 5 H z , 3 H , C M e ) . D e O - a c e t y l a t i o n w i t h m e t h a n o l i c sodium methoxide p r o v i d e d t i t l e compound 17 i n 89% y i e l d ; [ a ] - 8 2 ° (c 1 . 2 , H 0 ) , m / z : 5 7 0 . 2 (M + 1 ) , 5 9 2 . 2 (M + N a ) , 5 6 8 . 4 ( M - l ) " ; F o r C - N . M . R . , see T a b l e I . D

D

+

2

¥

1 3

A l l y l 0-(3-0-sulfo-B-D-galactopyranosyl) - (l-*3) - [ 0 (α-L-f u c o p y r a n o s y l ) - (l->4) ] -2-acetamido-2-deoxy-B-Dg l u c o p y r a n o s i d e sodium s a l t ( 2 4 ) . R e a c t i o n o f a l l y l 0 ( 2 , 3 , 4 , 6 - t e t r a - 0 - a c e t y l - B - D - g a l a c t o p y r a n o s y l - (l->3) - 2 acetamido-2-deoxy-B-D-glucopyranoside (4) with trimethylacetyl chloride (pivaloyl chloride) i n pyridine a f f o r d e d t h e 6 - 0 - p i v a l o y l compound 5 i n 90% y i e l d ; [ a ] + 1 3 ° ( c 1 . 0 , C H C I 3 ) ; H - N . M . R . ( C D C I 3 ) : δ 5 . 8 3 - 5 . 6 2 (m, 1 H , =CH) , 4 . 5 3 ( d , J = 8 . 5 H z , 1 H , H - l ) , 2.13-1.63 ( c l u s t e r o f s , 15 H , 4 χ OAc a n d N A c ) , 1 . 1 9 ( s , 9 H , CMe ) . D

1

1

3


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129

Synthetic Acceptors for OL-L-Fucosyltransferases

G l y c o s y l a t i o n of 5 with 2 under s i m i l a r c o n d i t i o n s a s d e s c r i b e d f o r t h e p r e p a r a t i o n o f 14 g a v e 18 i n 53% yield, [a] - 4 0 ° ( ç 1.0, CHC1 ) ; ^ - N . M . R . (CDC1 ) : δ 7 . 3 9 - 7 . 1 7 (m, 6 H , a r o m . ) , 6 . 7 8 - 6 . 6 9 (m, 6 H , a r o m . ) , 3 . 7 9 ( s , 9 H , 3 χ OMe) , 2 . 1 3 - 1 . 6 7 ( c l u s t e r o f s , 15 H , 4 x OAc a n d N A c ) , 1.23 ( s , 9 H , CMe ) , 1.07 ( d , J = 6 H z , 3 H , CMe). Removal o f t h e 4 - m e t h o x y b e n z y l group from compound 18 w i t h e e r i e ammonium n i t r a t e i n a c e t o n i t r i l e (48) p r o v i d e d compound 19 i n 59% y i e l d ; [ a ] - 7 1 ° (ç 1.0, CHC1 ); ^ - N . M . R . (CDCI3) : S 6.60 (d, J = 8.5 H z , 1 H , N H ) , 5 . 8 6 - 5 . 8 1 (m, 1 H , =CH), 5.38 ( d , J = 3.1 H z , 1 H , H - l " ) , 4.92 ( d , J = 8.1 H z , 1 H , H - l ) , 4 . 7 0 ( d , J = 6 . 5 H z , 1 H , H - l ) , 2 . 1 5 , 2 . 0 7 , 2 . 0 5 , 2.01 ( e a c h s , 8 H , 4 χ O A c ) , 1.97 ( s , 3 H , N A c ) , C - N . M . R . (CDCI3) : δ 9 9 . 9 9 ( Ο Ι ) , 98.7 ( C - l ) , 96.85 ( C - l " ) . D

3

3

3

D


3

1

1 3

1

The t r i o l 19 was c o n v e r t e d i n t o i t s 3",4"-(ethyl orthoacetate) which on d e - 0 - a c e t y l a t i o n followed by hydrolysis with 80% aqueous acetic acid gave the intermediate, a l l y l O-(B-D-galactopyranosyl)-(l->3)-[0-(4O - a c e t y l - a - L - f u c o p y r a n o s y l ) -(l-*4) ] - 2 - a c e t a m i d o - 2 - d e o x y - 6 O - p i v a l o y l - Ô - D - g l u c o p y r a n o s i d e (20) i n 46% y i e l d ; [ a ] 7 4 ° ( ç 1.0, CHCI3); ^ - N . M . R . (CDCI3 +CD3OD) : δ 5 . 9 0 - 5 . 8 0 (m, 1 H , =CH), 5 . 1 5 ( d , 1 H , H - 4 " ) , 4 . 8 5 ( d , J = 8.0 H z , 1 H , H - l ) , 2.14 ( s , 3 H , O A c ) , 1.98 ( s , 3 H , N A c ) , 1.22 (s, 9 H , C M e ) , 1.04 (d, J = 6.6 H z , 3 H , CMe); C N . M . R . (CDCI3 + CD3OD) : δ 102.7 ( C - l ' ) , 9 9 . 4 ( C - l ) , 97.5 ( C - l " ) , 27.0 (CMe ) . D

1

3

3

3

I s o p r o p y l i d e n a t i o n o f compound 20 u n d e r c o n d i t i o n s d e s c r i b e d by C a t e l a n i e t a l . (49) p r o v i d e d t h e 3 · , 4 ' acetal compound 21 i n 88% y i e l d , [a] - 4 4 ° (c 1.0, CHCI3); ^ H - N . M . R . (CDCI3) : δ 5 . 9 0 - 5 . 7 0 (m, 1 H , =CH) , 2.10 ( S , 3 H , O A c ) , 1.99 ( s , 3 H , N A c ) , 1.48 a n d 1.30 ( e a c h s , 6 H , CMe ) , 1.21 ( s , 9 H , CMe ) , 1 . 0 6 ( d , 3 H , CMe) . De-0-acetylation of compound 21 f o l l o w e d by a c e t y l a t i o n w i t h p y r i d i n e - a c e t i c a n h y d r i d e and h y d r o l y s i s o f i s o p r o p y l i d e n e w i t h 60% aqueous a c e t i c a c i d f u r n i s h e d t h e d i o l 22 i n 55% y i e l d ; [ a ] - 8 8 ° (c 0 . 3 , CHC1 ) . T h i s was t r a n s f o r m e d i n t o i t s 3 · , 4 ' ( e t h y l o r t h o a c e t a t e ) w h i c h was h y d r o l y z e d w i t h a c e t i c a c i d t o y i e l d a k e y intermediate, allyl 0-(2,4,6-tri-0-acetyl-B-Dg a l a c t o p y r a n o s y l ) - (l->3) - [0- (2 , 3 , 4 - t r i - O - a c e t y l - a - L f u c o p y r a n o s y l ) - ( 1-^4 ) ] - 2 - a c e t a m i d o - 6 - 0 - a c e t y l - 2 - d e o x y - B - D g l u c o p y r a n o s e 23; [ a ] - 6 5 ° (c 0 . 4 , CHC1 ) . S u l f a t i o n o f 23 with sulfurtrioxide-pyridine complex in N,Ndimethylformamide, followed by d e - 0 - a c e t y l a t i o n with m e t h a n o l i c s o d i u m m e t h o x i d e p r o v i d e d t h e t i t l e compound 24; [ a ] - 5 9 ° ( ç 0 . 6 , H 0 ) ; m / z ; 694.1 (M + N a ) , 6 4 8 . 3 (M-Na)"; F o r C - N . M . R . , see T a b l e I . D

2

3

D

D

3

3

+

D

2

1 3

Synthesis of benzyl 0-(3-0-sulfo-B-Dg a l a c t o p y r a n o s y l ) - ( i->3 ) - 2 - a c e t a m i d o - 2 - d e o x y - B - D g l u c o p y r a n o s i d e sodium s a l t ( 2 5 ) . The p r o c e d u r e s u s e d


130


f o r p r e p a r i n g 25 were e s s e n t i a l l y t h e same a s d e s c r i b e d for compound 13, except that benzyl 0-(B-Dg a l a c t o p y r a n o s y l ) - ( l-*3 ) - 2 - a c e t a m i d o - 2 - d e o x y - B - D g l u c o p y r a n o s i d e was u s e d a s t h e s t a r t i n g m a t e r i a l w h i c h was synthesized using the published procedure (50) . Compound 25 [ a ] - 1 2 ° (ç 0 . 9 , H 0 ) : m / z : 5 9 7 . 9 (M + N a ) , 5 5 2 . 1 ( M - N a ) ~ , 5 7 4 . 3 ( M - l ) ~ ; F o r C - N . M . R . , s e e T a b l e I.. +

D

2

1 3


Benzyl 0-(3-0-sulfo-B-D-galactopyranosyl-3)-(1-+4)-2acetamido-2-deoxy-B-D-glucopyranoside sodium s a l t (26)· Isopropylidenation of benzyl 0-(B-D-galactopyranosyl)(l-*4) - 2 - a c e t a m i d o - 2 - d e o x y - B - D - g l u c o p y r a n o s i d e (51) b y t h e p r o c e d u r e o f C a t e l a n i e t a l . (49) f o l l o w e d b y a c e t y l a t i o n with p y r i d i n e - a c e t i c anhydride afforded benzyl 0 - ( 2 , 6 - d i 0-acetyl-3,4-0-isopropylidene-B-D-galactopyranosyl) ( l->4 ) - 2 - a c e t a m i d o - 3 , 6 - d i - 0 - a c e t y l - 2 - d e o x y - B - D g l u c o p y r a n o s i d e i n 50% y i e l d ; [ a ] - 1 7 ° (ç 0 . 8 , CHCl^). Cleavage of the 3 ,4 -0-isopropylidene group with chloroform-trifluoroacetic acid-water, furnished a d i o l i n 92% y i e l d , [ a ] - 3 3 ° ( ç 0 . 7 , CHC1 ) . T h i s d i o l was converted into a 3,4-(ethyl orthoacetate) derivative, w h i c h was h y d r o l y z e d t o g i v e t h e i n t e r m e d i a t e , b e n z y l 0 (2,4,6-tri-0-acetyl-B-D-galactopyranosyl)-(1-+4)-2acetamido-3,6-di-0-acetyl-2-deoxy-B-D-glucopyranoside i n 81% y i e l d ; [ a ] - 4 0 ° ( ç 0 . 7 , CHC1 ) . T h i s compound o n t r e a t m e n t w i t h s u l f u r t r i o x i d e - p y r i d i n e complex i n N , N dimethylformamide followed by O-deacetylation with m e t h a n o l i c s o d i u m m e t h o x i d e p r o v i d e d t h e t i t l e compound 26 i n 55% y i e l d , [ a ] - 1 5 ° (ç 0 . 7 , H 0 ) , m / z : 5 9 8 . 0 (M + Na) , 552 (M - N a ) ~ , 574 (M-l)"; For C-N.M.R. , see Table I. D

1

1

D

3

D

3

D

+

2

1 3

Acknowledgment: T h e s e i n v e s t i g a t i o n s were s u p p o r t e d b y g r a n t N o s . CA35329 and CA16056 awarded b y t h e N a t i o n a l Cancer I n s t i t u t e .

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Synthetic Oligosaccharides - American Chemical Society

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