Synthesis of Novel Donor Mimetics of UDP-Gal, UDP-GlcNAc, and UDP-GalNAc as Potential Transferase Inhibitors

Article abstract of DOI:10.1021/jo990766l

For the enzymatic transfer of galactose, N-acetylglucosamine, and N-acetylgalactosamine, UDP-Gal (1), UDP-GlcNAc (2), and UDP-GalNAc (3) are employed, and UDP serves as a feedback inhibitor. In this paper the synthesis of the novel UDP-sugar analogues 4, 5, and 6 as potential transferase inhibitors is described. Compounds 4-6 feature C-glycosidic hydroxymethylene linkages between the sugar and nucleoside moieties in contrast to the anomeric oxygens in the natural derivatives 1-3.

Full text of DOI:10.1021/jo990766l

24
J . Org. Chem. 2000, 65, 24-29
Syn th esis of Novel Don or Mim etics of UDP -Ga l, UDP -GlcNAc, a n d
UDP -Ga lNAc a s P oten tia l Tr a n sfer a se In h ibitor s†
Andreas Scha¨fer and J oachim Thiem*
Institut fu¨r Organische Chemie, Universita¨t Hamburg, D-20146 Hamburg, Germany
Received May 7, 1999
For the enzymatic transfer of galactose, N-acetylglucosamine, and N-acetylgalactosamine, UDP-
Gal (1), UDP-GlcNAc (2), and UDP-GalNAc (3) are employed, and UDP serves as a feedback
inhibitor. In this paper the synthesis of the novel UDP-sugar analogues 4, 5, and 6 as potential
transferase inhibitors is described. Compounds 4-6 feature C-glycosidic hydroxymethylene linkages
between the sugar and nucleoside moieties in contrast to the anomeric oxygens in the natural
derivatives 1-3.
In tr od u ction
In recent years much progress has been made toward
a broader understanding of the functions of complex
oligosaccharide structures. It is now well accepted that
carbohydrates, in addition to their well-known role as
molecules for energy storage, are of crucial importance
in various biological processes such as cell-cell recogni-
tion, tumor cell metastasis, and leukocyte adhesion
during inflammation.1,2 Despite the rapidly growing
knowledge in this field, the molecular basis of many
processes is often still uncertain. The glycosyl trans-
ferases of the Leloir pathway are key catalysts for
synthesis of oligosaccharides and glycoconjugates in
vivo.3-5 These enzymes transfer activated monosaccha-
ride units in the form of their nucleotide diphosphate
derivatives to a specific free hydroxy group of the acceptor
molecule. Modulation or inhibition of this transfer reac-
tion provides an excellent opportunity for intervention
of the oligosaccharide biosynthesis and to obtain a more
complete understanding of the structure-function rela-
tion of oligosaccharides on a molecular basis.6
Because UDP-Gal (1), UDP-GlcNAc (2), and UDP-
GalNAc (3) are donor substrates of the various Gal-,
GlcNAc-, and GalNAc-transferases, we present herein
new donor mimetics 4-6 as potential inhibitors of the
corresponding transferases. Inhibition of these enzymes
is of particular interest, as they are involved in many
important biological processes. For example, UDP-Gal (1)
and UDP-GalNAc (3) are key intermediates in the
biosynthesis of O-glycoproteins, a process which is still
under investigation.7 UDP-Gal (1) and UDP-GlcNAc (2)
are intermediates in the biosynthesis of cell surface
ligands which mediate various cell-cell recognition
phenomena.1 UDP-GlcNAc (2) plays an important role
in the biosynthesis of N-glycoproteins in general and is
involved in the recently described dynamic O-GlcNAcyl-
ation of specific proteins.8,9 Inhibitors for these metabolic
pathways should be appreciated and enhance the under-
standing of such processes.10
A common approach for the synthesis of natural
saccharide phosphate mimetics is the substitution of a
phosphoester oxygen atom by a carbon atom, resulting
in the formation of isosteric C-glycosidic phosphonates.11-14
However, phosphonates have the inherent disadvantage
of being markedly different in charge and shape under
physiological conditions.15 In addition such ground-state
analogues are expected to show only modest inhibition
rates. For this reason we have decided to substitute the
(8) Schachter, H.; Brockhausen, I. In Glycoconjugates; Allen, H. J .,
Ed.; Marcel Dekker: New York, 1992; pp 263-332.
(9) Hart, G. W.; Kreppel; L. K.; Comer, F. I.; Arnold, C. S.; Snow,
D. M.; Ye, Z.; Cheng, X.; DellaManna, D.; Caine, D. S.; Earles, B. J .;
Akimoto, Y.; Cole, R. N.; Hayes, B. K. Glycobiology 1996, 6, 711-716.
(10) Sinnott, M. L. Chem. Rev. 1990, 90, 1171-1202.
(11) Schmidt, R. R.; Frische, K. Bioorg. Med. Chem. Lett. 1993, 3,
1747-1750.
(12) Casero, F.; Cipolla, L.; Lay, L.; Nicotra, F.; Panza, L. R.; Russo,
G. J . Org. Chem. 1996, 61, 3428-3432.
(13) J uan, J . I.; Gleason, J . G. Tetrahedron Lett. 1992, 33, 6911-
6914.
† Dedicated to Professor Pierre Sinay¨ on the occasion of his 62nd
birthday.
(1) Varki, A. Glycobiology 1993, 3, 97-130.
(2) Dwek, R. A. Chem. Rev. 1996, 96, 683-720.
(3) Kornfeld, S.; Kornfeld, R. Annu. Rev. Biochem. 1985, 54, 631-
664.
(4) Leloir, L. F. Science 1971, 172, 1299-1303.
(5) Leloir, L. F.; Caputto, R.; Cardini, C. E.; Paladini, A. C. J . Biol.
Chem. 1950, 184, 333-350.
(6) Amann, F.; Schaub, C.; Mu¨ller, B.; Schmidt, R. R. Chem. Eur.
J . 1998, 6, 1106-1115.
(14) Qiao, L.; Vederas, J . C. J . Org. Chem. 1993, 58, 3480-3482.
(15) Engel, R. Chem. Rev. 1976, 77, 349-367.
(7) Marth, J . D. Glycobiology 1996, 6, 701-705.
10.1021/jo990766l CCC: $19.00 © 2000 American Chemical Society
Published on Web 12/16/1999

Donor Mimetics of UDP-Gal, UDP-GlcNAc, and UDP-GalNAc
J . Org. Chem., Vol. 65, No. 1, 2000 25
Sch em e 1a
C-glycosidic linkages at the anomeric centers of GlcNAc
and GalNAc had to be established. In the area of
C-glycoside chemistry, only a limited number of proce-
dures are known for the demanding task to introduce a
C-glycosidic bond into 2-amino-sugars.18-21 We chose the
method recently published by Kessler et al. using a
dianion strategy.22-24 The benzylated, anomerically un-
blocked GlcNAc and GalNAc derivatives 12 and 13 were
converted to the anomeric chlorides according to the
literature.23 These were converted into their dilithium
intermediates, and by treatment with carbon dioxide the
1-C-R-carboxylic acids 14 and 15 were obtained. Subse-
quent methylation led to the methyl esters 16 and 17,
which were reductively converted to the 1-C-R-hydroxy-
methylene-GlcNAc and -GalNAc derivatives 18 and 19.
Attempts to form the 1-C-R-hydroxymethylene deriva-
tives directly by reaction of the dianion intermediates
with paraformaldehyde or formaldehyde gave no results
or only very low yields of the desired products. Phospho-
rylation of the benzylated 1-C-R-hydroxymethylene-
GlcNAc 18 gave the diphenyl phosphate 20, which could
be deprotected by hydrogenolysis to give the free phos-
phate 22, using the same procedure as described for the
galactose phosphate 11. To obtain the appropriate C-
glycosidic GalNAc phosphate, a different approach had
to be used, since all attempts to reductively cleave the
phenyl phosphate ester group of 21 accordingly did not
meet with success. Thus, starting with the alcohol 19,
the dibenzyl phosphite 23 was formed and oxidatively
converted to the dibenzyl phosphate 24. The following
hydrogenolysis using Pd/C gave the desired completly
deprotected phosphate 25 (Scheme 2).
a
Key: (a) Propargyl-TMS, CH3CN, BF3‚Et2O. (b) O3, CH2Cl2.
(c) NaBH4, EtOH. (d) PO(OPh)2Cl, Py, DMAP. (e) (1) Pd/C, MeOH.
(2) PtO2, MeOH, uridine 5′-monophosphomorpholidate, 1H-tetra-
zole, Py.
anomeric hydroxy group of Gal, GlcNAc, and GalNAc
with a hydroxymethylene group to give the homologous
C-glycosidic UDP-sugars 4-6 after phosphorylation and
coupling with UMP. Compounds 4-6 are distinguished
by phosphoester bonds between the sugars and the
nucleotides. The labile glycosidic phosphoester bonds of
the natural donors 1-3 are replaced by C-glycosidic
linkages to give 4-6 a high resistance to chemical and
enzymatic hydrolysis. The distances between the ano-
meric centers and the nucleotides are somewhat in-
creased. This may lead to stronger binding since the
compounds will more closely resemble the transition state
formed during the enzymatic reaction.16 Therefore, the
mimetics 4-6 may be considered to be suitable trans-
ferase inhibitors.
The synthesis of the UDP-Gal (4) UDP-GlcNAc (5), and
UDP-GalNAc (6) mimetics was performed following a
recent procedure of Wittmann and Wong,25 which is an
improved variant of the Moffat-Khorana phosphomor-
pholidate coupling method.26 The diphosphates could be
isolated in 48-55% yield. The 1H NMR and 13C NMR
spectra showed the expected signals; a nearly complete
assignment could be performed by 2D NMR experiments.
In the case of 6 signal broadening due to coalescence
phenomena prevented a detailed signal identification in
1
the H NMR spectrum. The 31P NMR spectra of 4, 5, and
Resu lts a n d Discu ssion
6 show the two characteristic phosphorus atom signals
with P-P geminal couplings. In the case of 6, low-
temperature spectra recorded at 253 K provided signals
with the expected doublet structures.
In this paper we report on the synthesis of C-glycosidic
UDP-Gal, UDP-GlcNAc, and UDP-GalNAc mimetics.
This novel class of compounds is presumed to feature
potent inhibition of glycosyltransferases because they are
structurally related to the transition states of the ap-
For the synthesis of the UDP-sugar mimetics 4-6 the
appropriate phosphates 11, 22, and 25 had to be synthe-
sized first. Starting from benzyl-protected methyl galac-
toside 7, conversion under Lewis acid catalysis (BF3‚
Et2O) gave the C-glycosidic R-allene derivative 8.17 Only
a minor amount of the â-anomer was formed, which could
easily be separated by column chromatography. Oxida-
tive cleavage of 8 by ozonolysis led to the appropriate
aldehyde, which was subjected to NaBH4 reduction to
give the C-glycosidic 1-hydroxymethylene derivative 9.
The subsequent phosphorylation was performed by treat-
ment of the alcohol 9 with diphenylchlorophosphate to
give the phosphate 10. Hydrogenolysis of the protective
groups using subsequently Pd/C and PtO2 afforded the
deprotected 1-C-R-hydroxymethylene galactose phos-
phate (12) (Scheme 1).
(18) Levy, E. D.; Tang, C. In The chemistry of C-glycosides; Baldwin,
J . E., Magnus, P. D., Eds.; Pergamon: Oxford, U.K., 1995.
(19) Carcano, M.; Nicotra, F.; Panza, L.; Russo, G. J . Chem. Soc.,
Chem. Commun. 1989, 297-299.
(20) Giannis, A.; Sandhoff, K. Carbohydr. Res. 1987, 171, 201-210.
(21) Grondin, R.; Leblanc, Y. Hoogsteen, K. Tetrahedron Lett. 1991,
32, 5021-5024.
(22) Kessler, H.; Hoffmann, M. Tetrahedron Lett. 1996, 35, 6067-
6070.
(23) Hoffmann, M.; Burkhart, F.; Hessler, G.; Kessler, H. Helv.
Chim. Acta 1996, 79, 1519-1532.
For access to the 1-C-R-hydroxymethylene-GlcNAc and
1-C-R-hydroxymethylene-GalNAc phosphates (22, 25)
(24) Burkhardt, F.; Kessler, H. Tetrahedron Lett. 1998, 39, 255-
256.
(25) V. Wittmann, V.; Wong, C. H. J . Org. Chem. 1997, 62, 2144-
2147.
(26) Moffat, J . G.; Khorana, H. G. J . Am. Chem. Soc. 1961, 83, 649-
658.
(16) Schramm, V. L. Annu. Rev. Biochem. 1998, 67, 693-720.
(17) Kobertz, W. R.; Bertozzi, C. R.; Bednarski, M. D. Tetrahedron
Lett. 1992, 33, 737-740.

26 J . Org. Chem., Vol. 65, No. 1, 2000
Scha¨fer and Thiem
Sch em e 2a
a
Key: (a) SOCl2, Tol. (b) (1) BuLi, THF. (2) LiNaph, THF. (3) CO2. (c) MeI, DMF. (d) NaBH4, EtOH. (e) PO(OPh)2Cl, Py, DMAP. (f) (1)
Pd/C, MeOH. (2) PtO2, MeOH. (g) Di(benzyloxy)-N,N-diisopropylphosphoamidite, CH2Cl2, 1H-tetrazole. (h) MCPBA. (i) Pd/C, MeOH. (j)
Uridine 5′-monophosphomorpholidate, 1H-tetrazole, Py.
137.26, 137.31 (24C, Ar). Anal. Calcd for C35H38O6: C, 75.79;
H, 6.91. Found: C, 75.58; H, 6.84.
propriate transfer reactions. Insight into various mech-
anisms of several metabolic processes is expected, bio-
logical evaluations are currently under investigation, and
the results will be reported in due course.
2,6-An h yd r o-3,4,5,7-t et r a -O-b en zyl-D-glycer o-L-glu co-
h ep titol-1-yl) Di-O-p h en yl P h osp h a te (10). Compound 9
(252 mg, 0.45 mmol) was dissolved in dry pyridine (3 mL)
under argon atmosphere, diphenyl chlorophosphate (0.2 mL,
1.0 mmol) and a catalytic amount of DMAP were added, and
the solution was stirred overnight. The solvent was evaporated
under reduced pressure and the residue codistilled three times
with toluene and purified by flash chromatography over silica
gel (petroleum ether (50-70)-EtOAc, 4:1) to give 273 mg (77%)
of the phosphate 10 as an oil: [R]20D ) +16.3 (c 1, CH2Cl2); 1H
NMR (CDCl3, 400 MHz) δ 3.59 (dd, 1H, J ) 4.6, 10.2, H-7),
3.60 (dd, 1H, J ) 3.6, 6.6, H-4), 3.70 (dd, 1H, J ) 7.1, 10.2,
H-7′), 3.80 (dd, 1H, J ) 4.1, 6.6, H-3), 3.93 (dd, 1H, J ) 3.6,
3.1, H-5), 4.09 (ddd, 1H, J ) 3.1, 4.6, 7.1, H-6), 4.17 (m, 1H,
H-2), 4.31 (m, 1H, H-1), 4.60 (m, 9H, 4 × CH2-Bn, H-1′), 7.25-
7.06 (m, 30H, Ar); 13C NMR (CDCl3, 100.62 MHz) δ 65.09 (d,
1C, J ) 6.1, C-1), 65.78 (1C, C-7), 68.91 (1C, C-2), 71.83, 72.12,
72.18, 72,45 (4 × 1C, CH2-Bn), 72.26 (1C, C-6), 72.75 (1C,
C-5), 74.02 (1C, C-3), 75.49 (1C, C-4), 119.05, 119.10, 119.14,
124.31, 126.44, 126.60, 126.75, 126.81, 126.93, 127.30, 127.36,
127.41, 128.73, 128.76, 137.23, 137.30 (36 C, Ar); 31P NMR
(CDCl3, 80 MHz) δ 1.0; MALDI-TOF (positive mode, DHB) m/z
calcd for C47H47O9P 786, found 809 (M + Na)+. Anal. Calcd
for C47H47O9P: C, 71.74; H, 6.02. Found: C, 71.28; H, 5.98.
Exp er im en ta l Section
Melting points are uncorrected. 1H NMR and 13C NMR
spectra were recorded on a Bruker AMX 400 or DMX 500. J
values are given in hertz. 2D experiments were done for all
novel compounds. Optical rotations were measured with a
Perkin-Elmer 243 polarimeter. MALDI-TOF mass spectra
were obtained on a Bruker Biflex III spectrometer. Merck silica
gel 60 (partical size 40-63 µm) was employed for flash
chromatography. For gel permeation chromatography Biogel
P-2 was used.
2,6-An h yd r o-3,4,5,7-t et r a -O-b en zyl-D-glycer o-L-glu co-
h ep titol (9). A solution of allene 817 (2.0 g, 3.55 mmol) in dry
CH2Cl2 (80 mL) was stirred at -65 °C. Ozone was bubbled
through for 2 h until the blue color indicated a saturated
solution. Excess ozone was removed by purging with nitrogen.
Dimethyl sulfide was added, and the solution was stirred
overnight. Evaporation under reduced pressure gave the crude
aldehyde, which was redissolved in dry ethanol (40 mL). The
solution was cooled to 0 °C, and sodium borohydride (0.3 g,
7.9 mmol) was added and stirred for 4 h. The mixture was
diluted with saturated aqueous NH4Cl and extracted with CH2-
Cl2. The organic phase was dried (MgSO4) and evaporated and
the residue chromatographically purified by flash chromatog-
raphy over silica gel (petroleum ether (50-70)-EtOAc, 4:1)
(2,6-An h ydr o-D-glycer o-L-glu co-h eptitol-1-yl) P h osph ate
(11). To a solution of di-O-phenyl phosphate 10 (263 mg, 0.33
mmol) in dry methanol (25 mL) was added Pd/C (10%, 100
mg), and the mixture was stirred for 72 h at 45 °C under
hydrogen atmosphere (1 bar). The catalyst was filtered off, and
the solvent was evaporated under reduced pressure. The oily
residue was redissolved in dry methanol (15 mL), PtO2 (50 mg)
was added, and the mixture was stirred for another 72 h under
hydrogen atmosphere. Filtration and evaporation afforded 82
mg (100%) of 11 as a white hygroscopic solid, which was used
to give alcohol 9 as an oil (672 mg, 34%): [R]20 ) +30.0 (c
D
1,CH2Cl2); 1H NMR (CDCl3, 400 MHz) δ 3.54 (dd, 1H, J ) 4.5,
10.2, H-1), 3.61 (dd, 1H, J ) 4.6, 11.7, H-7), 3.68 (dd, 1H, J )
3.1, 7.1, H-4), 3.75 (dd, 1H, J ) 8.1, 11.7, H-7′), 3.77-3.81 (m,
2H, H-1′, H-3), 3.92 (dd, 1H, J ) 3.1, 4.1, H-5), 3.97-4.03 (m,
2H, H-2, H-6), 4.36-4.65 (m, 8H, 4 × CH2-Bn), 7.16-7.27 (m,
20H, Ar), 13C NMR (CDCl3, 100.62 MHz) δ 59.78 (1C, C-7),
66.39 (1C, C-1), 70.30, 72.10 (2C, C-2, C-6), 71.98, 72.13, 72.27,
72.31 (4 × 1C, 4 × CH2-Bn), 73.00 (1C, C-5), 74.98 (1C, C-3),
75.55 (1C, C-4), 126.50, 126.64, 126.70, 126.75, 126.81, 126.89,
126.97, 127.00, 127.10, 127.33, 127.37, 127.48, 136.80, 137.15,
for the following conversion without further purification: [R]20
D
) +30.8 (c 0.5, H2O); 1H NMR (D2O, 400 MHz) δ 3.53 (dd, 1H,
J ) 4.1, 11.7, H-7), 3.60 (dd, 1H, J ) 7.6, 11.7, H-7′), 3.69 (dd,
1H, J ) 3.6, 9.7, H-4), 3.81-3.83 (m, 2H, H-5, H-6), 3.88 (dd,
1H, J ) 5.6, 9.7, H-3), 3.93 (dd, 1H, J ) 6.1, 8.6, H-1), 4.05-

Donor Mimetics of UDP-Gal, UDP-GlcNAc, and UDP-GalNAc
J . Org. Chem., Vol. 65, No. 1, 2000 27
4.09 (m, 2H, H-1′, H-2); 13C NMR (D2O, 100.62 MHz) δ 61.23
(1C, C-7), 61.75 (d, 1C, J ) 5.1, C-1), 67.86 (1C, C-3), 69.03
(15C, Ar), 169.94, 171.35 (2 × 1C, CONH, COOH); MALDI-
TOF (positive mode, DHB) m/z calcd for C30H33NO7 519, found
542 (M + Na)+ and 558 (M + K)+. Anal. Calcd for C30H33NO7:
C, 69.35; H, 6.40; N, 2.70. Found: C, 69.33; H, 6.48; N, 2.68.
3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-D-
glycer o-D-id o-h ep ton ic Acid Meth yl Ester (16). To a solu-
tion of acid 14 (3.45 g, 6.64 mmol) in DMF (40 mL) were added
CsCO3 (2.6 g) and methyl iodide (0.5 mL, 8.0 mmol), and the
mixture was stirred for 14 h. The solvent was removed under
reduced pressure and the residue dissolved in EtOAc-water.
The organic layer was washed with saturated aqueous NaH-
CO3 solution and brine, dried (MgSO4), and filtered. The
solvent was evaporated under vacuum to give 16 (3.0 g, 85%;
(1C, C-5), 70.34 (1C, C-4), 73.51 (1C, C-6), 74.57 (1C, C-2); 31
P
NMR (D2O, 80 MHz) δ 0.96; MALDI-TOF (positive mode,
DHB) m/z calcd for C7H15O9P 274, found 297 (M + Na)+ and
313 (M + K)+.
Am m on iu m (2,6-An h yd r o-D-glycer o-L-glu co-h ep titol-1-
yl) (Ur id in e-5′-yl) Dip h osp h a te (4). To a solution of phos-
phate 11 (80 mg, 0.29 mmol) in water (4 mL) were added
pyridine (12 mL) and tri-N-octylamine (128 µL, 0.29 mmol).
The solvent was removed under reduced pressure, and the oily
residue was codistilled with dry pyridine (3 × 2 mL) and dried
under vacuum for 2 h. Subsequently 4-morpholine-N,N′-
dicyclohexylcarboxamidinium uridine 5′-monophosphomor-
pholidate (367 mg, 0.465 mmol) and 1H-tetrazole (65 mg, 0.92
mmol) were added under argon atmosphere, and the mixture
was dissolved in dry pyridine (1.5 mL) and stirred at room
temperature for 2 d. The solution was diluted with water (2
mL) and evaporated. The residue was suspended in 0.1 M
aqueous NH4HCO3 and extracted with ether to give a clear
solution. The solvent was evaporated under reduced pressure,
redissolved in water, and lyophilized. The solid residue was
purified on Biogel P-2 to yield (83 mg, 48%) 4 as a white solid:
mp 120 °C) as a white solid: [R]20 ) +41.4 (c 0.37, acetone);
D
1H NMR (CDCl3, 400 MHz) δ 1.71 (s, 3H, CH3), 3.58 (dd, 1H,
J ) 9.7, 4.6, H-6), 3.65 (s, 3H, COOCH3), 3.67-3.73 (m, 3H,
H-4, H-7, H-7′), 4.08 (dd, 1H, J ) 5.6, 9.7, H-5), 4.42-4.52 (m,
6H, H-2, H-3, 2 x CH2-Bn), 4.54-4.60 (m, 2H, CH2-Bn), 6.34
(d, 1H, J ) 9.2, NH), 7.14-7.29 (m, 15H, Ar); 13C NMR (CDCl3,
100.62 MHz) δ 22.22 (1C, CH3), 46.98 (1C, C-3) 51.28 (1C,
COO-CH3), 66.44 (1C, C-7), 69.01 (1C, C-2), 71.77, 72.02, 72.25
(3 × 1C, 3 × CH2-Bn), 73.19 (1C, C-6), 74.48, 74.66 (2 × 1C,
C-4, C-5), 126.71, 126.82, 126.97, 127.15, 127.38, 127.50,
127.53, 136.33, 136.55, 136.91 (15C, Ar), 168.73, 169.34 (2 ×
1C, CONH, COOCH3); MALDI-TOF (DHB, positive mode) m/ z
calcd for C31H35NO7 533, found 534 (M + H)+. Anal. Calcd for
[R]20 ) +27.0 (c 1, H2O); Rf 0.25 (i-PrOH/1 M NH4HCO3); 1H
D
NMR (D2O, 500 MHz) δ 3.53 (dd, 1H, J ) 7.5, 11.8, H-7), 3.61
(dd, 1H, J ) 4.4, 11.8, H-7′), 3.72 (dd, 1H, J ) 3.4, 9.8, H-4),
3.87-3.84 (m, 1H, H-5, H-6), 3.88 (dd, 1H, J ) 5.6, 9.8, H-3),
4.02 (dd, 1H, J ) 5.5, 8.3, H-5-Rib), 4.04 (dd, 1H, J ) 5.7, 9.1,
H-1), 4.08-4.06 (m, 1H, H-5′-Rib), 4.10 (m, 1H, H-2), 4.14-
4.12 (m, 2H, H-4-Rib, H-1′), 4.22-4.19 (m, 2H, H-2-Rib, H-3-
Rib), 5.80 (d, 1H, J ) 4.1, H-1-Rib), 5.82 (d, 1H, J ) 8.1, H-5-
U), 7.80 (d, 1H, J ) 8.1, H-6-U);13C NMR (D2O, 125 MHz) δ
61.15 (1C, C-7), 62.60 (d, 1C, J ) 5.0, C-1), 65.27 (d, 1C, J )
5.1, C-5-Rib), 67.95 (1C, C-3), 69.00 (1C, C-5), 70.37 (1C, C-2-
Rib/C-3-Rib), 73.61 (1C, C-6), 73.72 (1C, C-4), 74.15 (1C, C-2-
Rib/C-3-Rib), 74.50 (1C, C-2), 83.63 (d, 1C, J ) 9.2, C-4-Rib),
88.69 (1C, C-1-Rib), 103.03 (1C, C-6-U), 142.02 (1C, C-5-U),
152.15 (1C, C-4-U), 166.63 (1C, C-2-U); 31P NMR (D2O, 80
MHz) δ -10.54 (d, 1P, J ) 20), -10.92 (d, 1P, J ) 20); MALDI-
TOF (negative mode, DHB) m/z calcd for C16H32N4O17P2 614,
C
31H35NO7: C, 69.78; H, 6.61; N, 2.62. Found: C, 69.65; H,
6.57; N, 2.59.
3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-1-
h yd r oxy-D-glycer o-D-id o-h ep titol (18). To a solution of 16
(3.0 g 5.62 mmol) in dry ethanol/THF (2.5:1, 10 mL) at 0 °C
was added sodium borohydride (638 mg, 10.87 mmol), and the
mixture was stirred for 0.5 h at this temperature and then 2
h at room temperature. The solvent was evaporated under
reduced pressure, and the residue was diluted with saturated
aqueous NH4HCO3 solution and CH2Cl2. The organic layer was
separated and the aqueous phase extracted with CH2Cl2 (2 ×
20 mL). The combined organic phases were washed with
saturated aqueous NH4HCO3 solution and brine, dried (Mg-
SO4), filtered, and evaporated under vacuum. Purification of
the crude product by flash chromatography over silica gel
found 579 (M - 2 NH4 + H+)-.
+
3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-D-
glycer o-D-id o-h ep ton ic Acid (14). To a suspension of 2-ac-
etamido-3,4,6-tri-O-benzyl-2-deoxy-R-D-glucopyranose (12)23
(5.0 g, 10.2 mmol) under argon atmosphere in dry toluene (30
mL) and dry chloroform (30 mL) was added thionyl chloride
(30 mL), and the resulting solution was stirred for 0.5 h.
Subsequently the solution was evaporated, and the solid
residue was codistilled with toluene (2 × 30 mL) and dried
under vacuum. The chloride was dissolved in dry THF (80 mL)
and cooled to -95 °C. Butyllithium (8 mL, 1.6 M in hexane,
12.8 mmol) was added (0.5 min) followed by a freshly prepared
1 M lithium naphthalenide solution (24 mL, 24 mmol) in THF
(0.5 min). Carbon dioxide was bubbled through for 1 h at -78
°C, and then the solution was warmed to room temperature
and diluted with saturated aqueous NH4HCO3 solution (30
mL) and EtOAc (50 mL). The organic phase was separated
and the aqueous phase acidified with 5 M aqueous HCl to pH
5 and extracted with EtOAc (3 × 30 mL). The combined
organic phases were dried (MgSO4), filtered, evaporated under
vacuum, and chromatographically purified over silica gel
(petroleum ether (50-70)-EtOAc, 2:1 to 1:3, and then EtOAc-
MeOH-AcOH, 20:2:1 to 2:2:1) to afford 14 (3.48 g, 66%) as a
yielded the alcohol 18 (1.82 g, 64%; mp 89-91 °C) as a white
1
solid: [R]20 ) +20.1 (c 1.22, acetone); H NMR (CDCl3, 400
D
MHz) δ 1.82 (s, 3H, CH3), 3.25 (dd, 1H, J ) 6.1,11.7, H-1),
3.52 (dd, 1H, J ) 9.1, 11.7, H-1′), 3.56 (d, 1H, J ) 3.0, H-4),
3.64 (d, 1H, J ) 3.0, H-5), 3.66 (dd, 1H, J ) 7.1, 10.2, H-7),
3.78 (dd, 1H, J ) 7.6, 10.2, H-7′), 3.96 (ddd, 1H, J ) 6.1, 8.7,
9.1, H-2), 4.15-4.22 (m, 2H, H-3, H-6), 4.37 (dd, 2H, CH2-
Bn), 4.44 (dd, 2H, CH2-Bn), 4.56 (dd, 2H, CH2-Bn), 7.13-
7.30 (m, 15H, Ar); 13C NMR (CDCl3, 100.62 MHz) δ 22.08 (1C,
CH3), 44.54 (1C, C-3) 59.83 (1C, C-1), 66.43 (1C, C-7), 66.66
(1C, C-2), 70.87, 70.92 (2 × 1C, 2 × CH2-Bn), 72.20 (1C, C-4),
72.33 (1C, CH2-Bn), 72.36 (1C, C-5), 73.56 (1C, C-6), 126.49,
126.68, 126.75, 126.85, 126.99, 127.29, 127.53, 127.67, 136.01,
136.18, 136.99 (15C, Ar), 170.50 (1C, CONH); MALDI-TOF
(positive mode, DHB) m/ z calcd for C30H35NO6 505, found 506
(M + H)+ and 528 (M + Na)+. Anal. Calcd for C30H35NO6: C,
71.27; H, 6.98; N, 2.77. Found: C, 71.34; H, 7.01; N, 2.74.
(3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-
D-glycer o-D-ido-h eptitol-1-yl) Di-O-ph en yl P h osph ate (20).
The procedure described for the synthesis of 10 was followed
with the alcohol 18 (240 mg, 0.48 mmol). Purification by flash
chromatography over silica gel (petroleum ether-EtOAc, 1:2)
afforded 303 mg of the phosphate 20 (86%) as a yellow oil:
yellow foam: [R]20 ) +28.3 (c 1.02, acetone); Rf 0.41 (CH2-
D
Cl2/MeOH/TFA, 3:1:0.05); 1H NMR (DMSO-d6, 400 MHz) δ
1.79 (s, 3H, CH3), 3.38 (dd, 1H, J ) 8.7, 7.6, H-5), 3.50 (d, 1H,
J ) 2.9, 11.2, H-7), 3.53 (dd, 1H, J ) 5.3, 11.2, H-7′), 3.78 (dd,
1H, J ) 7.6, 9.7, H-4), 3.88 (ddd, 1H, J ) 2.9, 5.3, 8.7, H-6),
3.97 (ddd, 1H, 5.8, 8.7, 9.3, H-3), 4.20 (d, 1H, J ) 5.8, H-2),
4.38-4.67 (m, 6H, 3 × CH2-Bn), 7.08-7.24 (15H, Ar), 7.80
(d, 1H, J ) 8.7, NH); 13C NMR (DMSO-d6, 100.62 MHz) δ 22.91
(1C, CH3), 50.59 (1C, C-3), 68.94 (1C, C-7), 72.57 (1C, CH2-
Bn), 72.74 (1C, C-2), 73.83 (1C, CH2-Bn), 74.00 (1C, CH2-
Bn), 74.92 (1C, C-6), 77.85 (1C, C-5), 79.12 (1C, C-4), 127.81,
127.94, 128.00, 128.06, 128.56, 128.63, 138.50, 138.56, 138.97
[R]20 ) -0.8 (c 0.2, CH2Cl2); 1H NMR (CDCl3, 400 MHz) δ
D
1.71 (s, 3H, CH3), 3.57-3.60 (m, 2H), 3.60 (dd, 1H, J ) 6.6,
9.7, H-7), 3.68 (dd, 1H, J ) 7.4, 9.7, H-7′), 4.11-4.25 (m, 5H),
4.34-4.41 (m, 4H, 2 × CH2-Bn), 4.50 (dd, 2H, CH2-Bn), 6.58
(d, 1H, J ) 9.67, NH), 7.04-7.26 (m, 25H, Ar); 13C NMR
(CDCl3, 100.62 MHz) δ 22.13 (1C, CH3), 44.99 (1C, C-3), 68.31
(d, 1C, J ) 4.1, C-1), 70.98, 71.18, 72.30 (3 × 1C, 3 × CH2-
Bn), 66.94, 72.04, 73.23, 73.99, 119.07, 119.12, 119.30, 123.36,
124.31, 126.57,126.59, 126.66, 126.76, 126.94, 127.12, 127.38,
127.51, 127.56, 128.36, 128.72, 136.23, 136.33 (30C, Ar), 169.12
(1C, CONH); MALDI-TOF (positive mode, DHB) m/ z calcd for

28 J . Org. Chem., Vol. 65, No. 1, 2000
Scha¨fer and Thiem
C42H44NO9P 737, found 738 (M + H)+. Anal. Calcd for C42H44
-
77.10 (1C, C-4), 128.19, 128.26, 128.33, 128,39, 128.95, 128.99,
129.24, 139.36, 139.48, 139.52 (15C, Ar), 170.43, 170.82 (2 ×
1C, CONH, COOCH3); MALDI-TOF (positive mode, DHB) m/ z
calcd for C31H35NO7 533, found 534 (M + H)+ and 556 (M +
Na)+. Anal. Calcd for C31H35NO7: C, 69.78; H, 6.61; N, 2.62.
Found: C, 69.52; H, 6.45; N, 2.60.
3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-1-
h yd r oxy-D-glycer o-L-glu co-h ep titol (19). The procedure
described for the synthesis of 18 was followed using the ester
NO9P: C, 68.38; H, 6.01; N, 1.90. Found: C, 68.98; H, 5.81;
N, 1.91.
(3-Acetam ido-2,6-an h ydr o-3-deoxy-D-glycer o-D-ido-h ep-
titol-1-yl) P h osp h a te (22). The procedure described for the
synthesis of 11 was followed using the phosphate 20 (293 mg,
0.4 mmol) to provide 111 mg of 22 (93%) as a hygroscopic white
solid, which was used for the following conversion without
further purification: [R]20D ) +29 (c 0.5, H2O); 1H NMR (D2O,
400 MHz) δ 1.88 (s, 3H, CH3), 3.27 (dd, 1H, J ) 3.5, 9.2, H-5),
3.59 (dd, 1H, J ) 4.3, 12.1, H-7), 3.68 (dd, 1H, J ) 8.0, 12.1,
H-7′), 3.75 (dd, 1H, J ) 9.7, 3.5, H-4), 3.87-3.91 (m, 2H, H-5,
H-6), 3.95 (dd, 1H, J ) 5.7, 9.7, H-3), 4.00 (dd, 1H, J ) 5.8,
8.3, H-1), 4.10-4.16 (m, 2H, H-1′, H-2);13C NMR (D2O, 100.62
MHz) δ 24.84 (1C, CH3), 55.27 (1C, C-3), 63.99 (1C, C-7), 66.33
(d, 1C, J ) 5.1, C-1), 73.53 (1C, C-5), 74.21 (1C, C-4), 75.60
(1C, C-2), 77.69 (1C, C-6), 170.32 (1C, CONH); 31P NMR (D2O,
80 MHz) δ 0.57.
17 (1.2 g, 2.25 mmol) to afford 808 mg of 19 (71%; mp 139 °C)
1
as a white solid: [R]20 ) +31.6 (c 1, H2O); H NMR (CDCl3,
D
400 MHz) δ 1.90 (s, 3H, CH3), 3.49-3.56 (m, 2H, CH2-1), 3.65-
3.70 (m, 2H, H-5, H-7), 3.86 (dd, 1H, J ) 3.0, 4.5, H-4), 4.02
(m, 1H, H-2), 4.08 (dd, 1H, J ) 9.0, 11.7, H-7′), 4.14 (ddd, 1H,
J ) 4.5, 4.5, 7.8, H-3), 4.24 (ddd, 1H, J ) 2.8, 6.0, 9.0, H-6),
4.37-4.64 (m, 6H, 3 × CH2-Bn), 7.16-7.24 (m, 15H, Ar); 13
C
NMR (CDCl3, 100.62 MHz) δ 22.33 (1C, CH3), 48.96 (1C, C-3)
61.18 (1C, C-1), 64.61 (1C, C-7), 65.57 (1C, C-2), 70.20 (1C,
CH2-Bn), 71.60 (1C, C-5), 71.68 (1C, CH2-Bn), 72.28 (1C,
CH2-Bn), 73.04 (1C, C-4), 74.04 (1C, C-6), 126.53, 126.59,
126.62, 126.68, 126.82, 126.88, 127.32, 127.41, 136.68, 137.13,
137.22 (15C, Ar), 169.57 (1C, CONH); MALDI-TOF (positive
mode, DHB) m/ z calcd for C30H35NO6 505, found 506 (M +
H)+ and 528 (M + Na)+. Anal. Calcd for C30H35NO6: C, 71.27;
H, 6.98; N, 2.77. Found: C, 70.98; H, 6.91; N, 2.72.
Am m on iu m (3-Aceta m id o-2,6-a n h yd r o-3-d eoxy-D-glyc-
er o-D-id o-h ep titol-1-yl) (Ur id in e-5′-yl) Dip h osp h a te (5).
The procedure described for the synthesis of 4 was followed
using 22 (100 mg, 0.32 mmol) to give 107 mg of 5 (53%) as a
white solid: [R]20 ) +30.6 (c 1, H2O); Rf 0.24 (i-PrOH/1 M
D
NH4HCO3) 1H NMR (D2O, 500 MHz) δ 1.89 (s, 3H, CH3), 3.29
(dd, 1H, J ) 8.2, 9.9, H-5), 3.59 (dd, 1H, J ) 5.2, 12.3, H-7),
3.76 (dd, 1H, J ) 2.3, 12.3, H-7′), 3.80 (ddd, 1H, J ) 2.3, 5.2,
9.9, H-6), 3.90 (dd, 1H, J ) 8.2, 10.6, H-4), 3.95 (dd, 1H, J )
6.0, 10.6, H-3), 4.03-4.09 (m, 3H, H-1, H-2, H-5′-Rib), 4.12
(dd, 1H, J ) 4.5, 11.8, H-5-Rib), 4.15-4.19 (m, 2H, H-1, H-4-
Rib), 4.23-4.26 (m, 2H, H-2-Rib, H-3-Rib), 5.85 (d, 1H, J )
4.1, H-1-Rib), 5.86 (d, 1H, J ) 8.2, H-5-U), 7.83 (d, 1H, J )
8.2, H-6-U); 13C NMR (D2O, 125 MHz) δ 22.32 (1C, CH3), 52.73
(1C, C-3), 61.47 (1C, C-7), 65.12 (d, 1C, J ) 6.1, C-5-Rib), 65.30
(d, 1C, J ) 5.1, C-1), 70.05, 71.03 (1C, C-5), 72.91 (d, 1C, J )
9.2, C-2), 71.73 (1C, C-4), 74.15, 75.42 (1C, C-6), 83.58 (1C,
C-4-Rib), 88.76 (1C, C-1-Rib), 103.02 (1C, C-6-U), 141.99 (1C,
C-5-U), 152.20 (1C, C-4-U), 166.61 (1C, C-2-U), 175.19 (1C,
CONH); 31P NMR (D2O, 80 MHz) δ -10.84 (d, 1P, J ) 20.9),
(3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-
D-glycer o-L-glu co-h ep t it ol-1-yl) Di-O-b en zyl P h osp h a t e
(24). To a stirred suspension of 1H-tetrazole (167 mg, 2.38
mmol) in dry CH2Cl2 (1.5 mL) was added dibenzyl-N,N-
diisopropylphosphoramidite (0.39 mL, 1.19 mmol), and the
mixture was stirred for 15 min under argon atmosphere. The
alcohol 19 (240 mg, 0.48 mmol), dissolved in dry CH2Cl2 (1.5
mL), was added, and the mixture was stirred at room tem-
perature for 4 h. Subsequently the reaction mixture was cooled
to 0 °C, MCPBA (70%, 330 mg, 1.37 mmol) was added, and
stirring was continued for 45 min. The solvent was removed
under reduced pressure (bath temperature <40 °C), and the
residue was purified by flash chromatography over silica gel
(petroleum ether (50-70)-EtOAc, 1:1 to 1:2) to afford 265 mg
-11.24 (d, 1P, J ) 20.9); MALDI-TOF (negative mode, DHB)
of the phosphate 24 (74%) as an oil: [R]20 ) +4.6 (c 1,CH2-
+
D
m/ z calcd for C18H35N5O17P2 655, found 620 (M - 2NH4
+
Cl2); 1H NMR (CDCl3, 500 MHz) δ 1.82 (s, 3H, CH3), 3.62 (dd,
1H, J ) 3.5, 11.4, H-7), 3.67 (dd, 1H, J ) 2.8, 5.5, H-5), 3.74
(dd, 1H, J ) 3.0, 5.5, H-4), 3.86-3.92 (m, 2H, H-1, H-7′), 4.04
(dd, 1H, J ) 7.2, 11.8, H-1′), 4.13 (dd, 1H, J ) 2.8, 3.5, H-6),
4.18 (dd, 1H, J ) 5.7, 7.2, H-2), 4.26 (m, 1H, H-3), 4.36-4.60
(m, 6H, 3 × CH2-Bn), 4.88-4.96 (m, 4H, 2 × CH2-Bn-PO4),
5.95 (d, 1H, J ) 8.3, NH), 7.15-7.23 (m, 25H, Ar); 13C NMR
(CDCl3, 100.62 MHz) δ 22.14 (1C, CH3), 47.66 (1C, C-3), 66.17
(d, 1C, J ) 6.1, C-1), 68.53, 68.61 (2C, 2 × CH2-Bn-PO4),
68.66 (1C, C-7), 70.89 (1C, CH2-Bn), 71.42 (1C, C-5), 71.56,
72.22 (2 × 1C, 2 × CH2-Bn), 73.76 (1C, C-4), 74.07, 74.10 (2
× 1C, C-2, C-6) 126.64, 126.67, 126.79, 126.91, 126.99, 127.36,
127.38, 127.56, 127.61, 127.67, 134.68, 136.74, 137.06, 137.19
(30C, Ar), 169.25 (1C, CONH); 31P NMR (CDCl3, 80 MHz,) δ
-0.04; MALDI-TOF (positive mode, DHB) m/ z calcd for
H)-.
3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-D-
glycer o-L-glu co-h ep ton ic Acid (15).27 Following the proce-
dure described for the synthesis of 14, compound 13 (2.24 g,
4.6 mmol)24 was converted to afford of 15 (1.24 g, 53%) as a
yellow foam: [R]20 ) +12.5 (c 0.5, CHCl3); Rf 0.40 (CH2Cl2/
D
1
MeOH/TFA, 3:1:0.05); H NMR (DMSO-d6, 400 MHz) δ 1.85
(s, 3H, CH3), 3.60 (dd, 1H, J ) 4.5, 10.6, H-7), 3.73 (m, 1H,
H-7′), 3.91 (dd, 1H, J ) 3.1, 7.6, H-4), 4.07 (dd, 1H, J ) 3.1,
3.1, H-5), 4.30 (m, 1H, H-6), 4.41-4.59 (m, 8H, H-2, H-3, 3 ×
CH2-Bn), 7.19-7.39 (m, 15H, Ar), 8.07 (d, 1H, J ) 7.6, NH);
13C NMR (DMSO-d6, 100.62 MHz) δ 22.88 (1C, CH3), 47.88
(1C, C-3), 67.61 (1C, C-7), 71.74 (1C, C-2), 72.41 (2 × 1C, 2 ×
CH2-Bn), 73.18 (1C, C-5), 74.08, 74.16 (2 × 1C, CH2-Bn, C-6),
76.03 (1C, C-4), 127.76, 127.84, 128.01, 128.47, 128.54, 128.56,
128.59, 138.67, 138.84, 138.86 (15C, Ar), 169.94, 171.32 (2 ×
1C, CONH, COOH); MALDI-TOF (positive mode, DHB) m/ z
calcd for C30H33NO7 519, found 520 (M + H)+ and 542 (M +
Na)+. Anal. Calcd for C30H33NO7: C, 69.35; H, 6.40; N, 2.70.
Found: C, 68.89; H, 6.31; N, 2.58.
C
44H48NO9P 765, found 766 (M + H)+ and 788 (M + Na)+.
Anal. Calcd for C44H48NO9P: C, 69.01; H, 6.32; N, 1.83.
Found: C, 68.83; H, 6.42; N, 1.78.
(3-Acet a m id o-2,6-a n h yd r o-3-d eoxy-D-glycer o-L-glu co-
h ep titol-1-yl) P h osp h a te (25). To a solution of 24 (250 mg,
0.33 mmol) in dry methanol (25 mL) was added Pd/C (10%,
125 mg), and the suspension was stirred under hydrogen
atmosphere at 50 bar for 10 h. The catalyst was filtered off,
and the solvent was removed under reduced pressure to afford
95 mg of 25 (91%) as a white hygroscopic solid, which was
3-Aceta m id o-2,6-a n h yd r o-4,5,7-tr i-O-ben zyl-3-d eoxy-D-
glycer o-L-glu co-h ep ton ic Acid Meth yl Ester (17). The
procedure described for the synthesis of 16 was followed using
15 (1.0 g, 1.93 mmol) to give 1.24 g of 17 (97%; mp 133 °C) as
a yellow solid: [R]20D ) +10.4 (c 0.2, CH2Cl2); 1H NMR (CDCl3,
400 MHz) δ 1.81 (s, 3H, CH3), 3.63-3.69 (m, 4H, CH3, H-6),
3.75-3.83 (m, 3H, H-4, H-7, H-7′), 4.02-4.07 (m, 1H, H-5),
4.39-4.66 (m, 6H, H-2, H-3, 3 × CH2-Bn), 6.68 (d, 1H, J )
7.6, NH), 7.19-7.28 (m, 15H, Ar); 13C NMR (100.62 MHz,
DMSO-d6, 363 K): δ 23.27 (1C, CH3), 48.68 (1C, C-3) 52.35
(1C, COO-CH3), 68.65 (1C, C-7), 72.26 (1C, C-2), 72.56, 73.20,
73.83 (3 × 1C, 3 × CH2-Bn), 74.21 (1C, C-6), 74.97 (1C, C-5),
used for the next conversion without further purification:
1
[R]20 ) +31.8 (c 1, H2O); H NMR (D2O, 400 MHz) δ 1.92 (s,
D
3H, CH3), 3.57 (dd, 1H, J ) 4.4, 11.8, H-7), 3.62 (dd, 1H, J )
7.8, 11.8, H-7′), 3.87 (dd, 1H, J ) 1.5, 3.3, H-5), 3.91-3.96 (m,
3H, H-1,H-4, H-6), 4.07 (dd, 1H, J ) 11.8, H-1), 4.11 (dd, 1H,
J ) 5.8, 6.5, H-2), 4.18 (dd, 1H, J ) 6.5, 10.9, H-3); 13C NMR
(D2O, 100.62) δ 22.32 (1C, CH3), 48.99 (1C, C-3), 61.81 (1C,
C-7), 63.82 (d, 1C, J ) 5.1, C-1), 68.31, 68.64 (1C, C-5), 72.98
(d, 1C, J ) 10.2, C-2), 74.20, 162.52 (1C, CONH); 31P NMR
(D2O, 80 MHz) δ 1.02.
(27) 15 is described in ref 24, but no analytical data were given.

Donor Mimetics of UDP-Gal, UDP-GlcNAc, and UDP-GalNAc
J . Org. Chem., Vol. 65, No. 1, 2000 29
Am m on iu m (3-Aceta m id o-2,6-a n h yd r o-3-d eoxy-D-glyc-
er o-L-glu co-h ep titol-1-yl) (Ur id in e-5′-yl) Dip h osp h a te (6).
The procedure described for the synthesis of 4 was followed
using 25 (90 mg, 0.29 mmol) to give 112 mg of 6 (55%) as a
(d, 1P, J ) 20.0), -8.73 (d, 1P, J ) 20.0); MALDI-TOF
(negative mode, DHB) m/ z calcd for C18H35N5O17P2 655, found
620 (M - 2 NH4 + H)-.
+
white solid: [R]20 ) +45.2 (c 1, H2O); Rf 0.24 (i-PrOH/1 M
Ack n ow led gm en t. Financial support of this study
by the Deutsche Forschungsgemeinschaft (SFB 470) and
the Fonds der Chemischen Industrie is gratefully ac-
knowledged.
D
NH4HCO3); 1H NMR (D2O, 500 MHz) δ 1.89 (2, 3H, CH3),
3.46-3.58 (m, 2H, CH2-7), 3.82 (m, 1H, H-5), 3.95-4.20 (m,
11H), 5.79-5.82 (m, 2H, H-1-Rib, H-5-U), 7.79 (d, 1H, J ) 7.9,
H-6-U); 13C NMR (D2O, 125 MHz) δ 22.38 (1C, CH3), 48.92
(1C, C-3), 61.90 (1C, C-7), 65.28 (d, 1C, J ) 7.12, C-5-Rib),
65.35, 68.29, 68.67, 70.03 (1C, C-2), 74.16, 74.45, 83.60 (d, 1C,
C-4-Rib), 88.78 (1C, C-1-Rib), 103.00 (1C, C-6-U), 142.00 (1C,
C-4-U), 152.20 (1C, C-4-U), 164.62 (1C, C-2-U), 175.73 (1C,
CONH); 31P NMR (D2O-MeOH, 2:1; 80 MHz, 253 K) δ -8.99
Su p p or tin g In for m a tion Ava ila ble: 13C NMR spectra for
4-6, 11, 22, and 25. This material is available free of charge
via the Internet at http://pubs.acs.org.
J O990766L