Synthesis of nonnatural nucleoside diphosphate sugars

Article abstract of DOI:10.1002/ejoc.201100906

Recently, we reported an efficient chemical method for the synthesis of a variety of naturally occurring nucleoside diphosphate (NDP) sugars. This method, which is based on the cycloSal approach, can also be used, in principle, for the preparation of rare or even nonnatural NDP sugars. Herein, the syntheses of sulfoquinovose-, glucose-6-sulfate-, L-galactose-, and 2-fluoroglycopyranoside- containing NDP sugars are presented, as well as the synthesis of NDP sugars with non-natural nucleosides. The reactions described gavestereoisomerically defined NDP sugars in high yields and short reaction times.

Full text of DOI:10.1002/ejoc.201100906

FULL PAPER
DOI: 10.1002/ejoc.201100906
Synthesis of Nonnatural Nucleoside Diphosphate Sugars
Saskia Wolf,[a] Rosmirt Molina Berrio,[a] and Chris Meier*[a]
Keywords: Sugar nucleotides / cycloSal approach / Carbohydrates / Sulfochinovose / Fluorinated sugars
Recently, we reported an efficient chemical method for the
synthesis of a variety of naturally occurring nucleoside di-
phosphate (NDP) sugars. This method, which is based on the
cycloSal approach, can also be used, in principle, for the
preparation of rare or even nonnatural NDP sugars. Herein,
ose-, and 2-fluoroglycopyranoside-containing NDP sugars
are presented, as well as the synthesis of NDP sugars with
non-natural nucleosides. The reactions described gave
stereoisomerically defined NDP sugars in high yields and
short reaction times.
the syntheses of sulfoquinovose-, glucose-6-sulfate-, L-galact-
transfer pathways, the energy-rich linkage of the anomeric
carbon atom and the β-phosphate of the NDP is cleaved to
release the NDP moiety as a side product.
Naturally occurring sugar nucleotides, as well as struc-
tural analogues, are of great interest as substrates for enzy-
matic reactions in carbohydrate synthesis, as enzyme inhibi-
tors, as tools for assay development, for the study of glyco-
conjugate biosynthesis, and for biosynthetic studies of oli-
gosaccharides (e.g., lipopolysaccharides).[4]
Introduction
Carbohydrates are one of the four major classes of bio-
molecules and form the largest amount of organic com-
pounds in nature. For example, they are used for energy
storage, as components of DNA or RNA backbones, as
structure elements in cell walls of bacteria, and they play a
key role in recognition processes. Essential intermediates in
carbohydrate metabolism and glycoconjugate biosynthesis
are sugar nucleotides, for example, nucleoside diphosphate
As structural analogues, modifications at the sugar
(NDP) sugars (Leloir pathway). Structurally, natural sugar moiety and the nucleoside are possible. As examples for
nucleotides (1) are composed of a sugar and a nucleoside modified sugar parts, several different variants are known.
mono- or diphosphate (Figure 1). Numerous combinations It is possible to use l-configured sugars instead of com-
of sugars and nucleotides are found in nature. Only nine monly occurring d-pyranoses. In addition, the plant sulfo-
different sugar nucleotides have been identified in mamma- lipid 1Ј,2Ј-di-O-acyl-3Ј-O-(6-deoxy-6-sulfo-α-d-glucopyr-
lian cells so far,[1] but the diversity of NDP sugars is much anosyl)-sn-glycerol is a membrane lipid found in higher
higher in other organisms. Beside their importance for the plants, cyanobacteria, and some other photosynthetic pro-
Leloir pathway,[2,3] sugar nucleotides serve as precursors of karyotes.[5] With regard to the sulfur budget of plants, the
deoxysugars, aminodeoxysugars, chain-branched sugars, sulfolipid represents a major component, which accounts
uronic acids, and glycoconjugates. In biosynthetic sugar- for about half of the organic sulfur in leaves.[6] It has been
Figure 1. General structure of sugar nucleotides 1.
suggested that a sulfo sugar nucleotide could be involved in
[a] Organic Chemistry, Department of Chemistry, Faculty of
biosynthesis. The availability of such sulfo sugar nucleotides
Sciences, University of Hamburg,
would enable the above-mentioned assumption to be inves-
tigated. Additionally, as a replacement for hydrogen, fluor-
ine atoms are attractive because of their high electronegati-
vity and metabolic stability. The application of fluorinated
Martin-Luther-King-Platz 6, 20146 Hamburg, Germany
Fax: +49-40-42838-5592
E-mail: chris.meier@chemie.uni-hamburg.de
Supporting information for this article is available on the
WWW under http://dx.doi.org/10.1002/ejoc.201100906.
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Synthesis of Nonnatural Nucleoside Diphosphate Sugars
sugars could be of great interest, because fluorinated com- tides[22] as starting materials. For the synthesis of NDP
pounds are used in biochemistry to improve the activity of sugars, these active phosphate esters were treated with an-
pharmaceutically relevant compounds.[7] Also, positron omerically pure glycopyranosyl phosphates as nucleophiles
emission tomography (PET) uses 2-fluoro-2-deoxy-d-glu- with the formation of the pyrophosphate bond.[23–25] This
cose as a fluorine-18 tracer in clinical oncology.[8]
approach led to NDP sugars in high yields, and it was
Furthermore, nucleoside analogues, instead of the natu- proven that the approach could be applied to various natu-
ral compounds, can also offer interesting applications in the ral sugars and to a variety of natural nucleosides. The first
inhibition of glycosyltransferases due to less selective re- examples of modified sugar moieties included the syntheses
cognition and binding to the enzyme. Selective binding to of cytidine diphosphate-6-deoxy-α- and -β-d-gulose and the
the enzyme is mainly influenced by the nucleoside heterocy- l-configured sugars thymidine diphosphate-α- and -β-l-fu-
cle, but also to a lesser degree by the sugar moiety of the cose (75%).[24] Herein, we focused our efforts on the synthe-
sugar nucleotide. Due to the influence of enzyme–substrate sis of NDP sugars with unnatural or rare sugars, for exam-
complexes in the biosynthesis of oligosaccharides, it may be ple, sulfo or sulfate groups and fluorinated sugars, or nu-
possible to control the elongation of oligosaccharide cleoside analogues.
chains.[2,3] For visualization processes, it is necessary to la-
bel the NDP sugar, for example, with (fluorescent) dyes or
even introduce a radiolabel. In addition, it may be possible
Results and Discussion
to deliver active antiviral or antitumor compounds into
cells using, for example, the mannose receptor.[9] This recep-
tor exists on the surface of macrophages and has a high
affinity for l-fucose and d-mannose at the terminus of oli-
gosaccharides.
The synthesis of NDP sugars by the cycloSal approach is
based on a convergent strategy. The key step is the coupling
between cycloSal nucleotides and anomerically pure glyco-
pyranosyl phosphates (Scheme 1).
Therefore, methods for the efficient preparation of natu-
ral and especially nonnatural sugar nucleotides are of con-
siderable importance for synthetic, biological, and medici-
nal chemistry. According to the Moffat–Khorana method,
glycosyl phosphates are coupled with nucleotide morpholi-
dates.[10,11] However, this reaction is normally slow, and the
yields are often very poor. Instead of morpholidates, imi-
dazolides have also been used without marked improvement
to the yields.[12–14] Van der Marel et al. published an ap-
proach in which the pyrophosphate group was formed by
using 5Ј-nucleoside phosphoramidites treated with sugar
phosphates.[15] The formed PIII–PV intermediate was oxid-
ized, and different protecting groups were cleaved. Alterna-
tively, the groups of Hindsgaul and Jakeman published a
procedure starting from glycopyranosyl bromides.[16,17]
However, the yields were only moderate and the stereo-
chemistry at the anomeric center could only be controlled
in the latter case due to the anchimeric effect of the protect-
ing groups. Similarly, Nugier-Chauvin et al. used unprotec-
ted thioimidoyl furanosides as starting materials for the
preparation of rare nucleotide furanoses.[18] The best yields
obtained were 37%, and again the stereochemistry at the
anomeric center could not be controlled. Thiem and co-
workers reported an enzymatic procedure starting from un-
protected sugars that were first phosphorylated and then
treated with a nucleoside triphosphate in a three-enzyme-
catalyzed pathway.[19,20] However, the chemical yields sel-
dom exceeded 30%, and the method entirely depended on
the availability of the necessary kinases and NDP sugar py-
Scheme 1. General principle of coupling with cycloSal nucleotides.
Purification can easily be achieved with chromatography
on RP-18 silica gel after cleavage of the acetyl groups,
which are used as protecting groups for the nucleosides and
sugar moieties. Preparations of cycloSal nucleotides and
glycopyranosyl phosphates were carried out as described
previously. The general coupling conditions have also been
described before.[25]
Synthesis of Sulfo- and Sulfate-Substituted NDP Sugars 4
and 6
To synthesize the target NPD sugars, 5-nitro-cycloSal-ur-
rophosphorylases, as well as on (sometimes) expensive nu- idine monophosphate (2) was prepared first in 93% yield.
cleoside triphosphates. In addition, an enzymatic approach The 31P NMR spectrum of the crude reaction product dis-
is limited to natural substrates or is at least dependent on played only the two expected resonance lines corresponding
the acceptance of the substrates by the enzymes used.
to the two diastereomers of the product.[24] A separation of
Recently, we reported a new chemical route to sugar nu- the diastereomers was not necessary, because the chirality
cleotides and other phosphorylated biomolecules, such as at the phosphorus atom was eliminated in the following re-
nucleoside triphosphates,[21] that used cycloSal nucleo- action with the sugar phosphate nucleophile.
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6-Deoxy-6-sulfo-α-d-glucopyranosyl phosphate (3) was diphosphate (UDP)-6-deoxy-6-sulfo-α-d-glucose (4) (UDP-
synthesized as reported before.[26,27] The obtained material α-d-sulfoquinovose) in a yield of 48% after cleavage of the
was completely unprotected at the hydroxy functions to give acetyl groups in the nucleoside part (Scheme 2). This
a tris(cyclohexylammonium) salt instead of the usually used proved that for the reaction of the sugar phosphates with
triethylammonium salt. Coupling of 2 and 3 formed uridine
1
Figure 2. 31P NMR spectrum of UDP-α-d-sulfoquinovose 4.
Figure 3. H NMR spectrum of UDP-α-d-sulfoquinovose 4.
Scheme 2. Synthesis of sulfo- and sulfate-substituted NDP sugars 4 and 6.
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Synthesis of Nonnatural Nucleoside Diphosphate Sugars
the cycloSal-triester no protection at the sugar hydroxy
The fluorine atoms were introduced by the addition of
groups was necessary. The 31P and 1H NMR spectra of 2 equiv. of Selectfluor (13) in an electrophilic fluorination
product 4 are shown in Figures 2 and 3, respectively.
of glycals 7 and 8 (Scheme 4).[8,29] In the case of galactal 8,
To compare the influence of sulfo and sulfate groups, we which has an axial 4-hydroxy group, the reaction proceeded
also synthesized UDP-α-d-glucose 5 (75% yield) and sul- highly selectively. Mechanistically, it is a syn addition to gal-
fated the 6-position. This reaction succeeded when using a actal. Due to steric effects, the reaction only took place
sulfur trioxide–pyridine complex in pyridine as the solvent from the opposite side of the axial acetate group, and, thus,
overnight at room temperature to give the product in quan- there was selective formation of the thermodynamically
titative yield (Scheme 2). The success of this reaction also more stable 4C1 conformation. After heterolytic bond disso-
demonstrates the possibility of carrying out additional syn- ciation to the energetically preferred carbocation, a water
thetic steps on NDP sugars.
molecule reacted with the positively charged carbon atom,
and products 15–17 were formed. In the case of glucal 7,
electrophilic fluorination was not selective because of the
missing assistance of the axial group; therefore, 2Ј-fluori-
Synthesis of Fluorinated NDP Sugars 24–26
For the synthesis of NDP sugars with a fluoro substitu- nated glucose and 2Ј-fluorinated mannose derivatives were
ent in the 2-position, first the glycals derived from glucose formed. β-Anomers of mannose compound 17 were not
and galactose were prepared (Scheme 3).[28] After peracety- found.
lation and bromination at the anomeric center, reduction
with zinc and N-methylimidazole led to glucal 7 in a yield ture overnight and led to 3,4,6-tri-O-acetyl-2-fluoro-2-de-
of 67% and to galactal 8 in a yield of 34%. oxy-α/β-d-galactose (15) in a yield of 30% and 3,4,6-tri-
The reaction was carried out in DMF at room tempera-
Scheme 3. Synthesis of glycals 7 and 8.
Scheme 4. Fluorination of glycals 7 and 8 with 13.
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O-acetyl-2-fluoro-2-deoxy-α/β-d-glucose (16) as well as the chromatographic steps. The α/β ratio was 1:0.4. After cleav-
3,4,6-tri-O-acetyl-2-fluoro-2-deoxy-α-d-mannose (17) (ratio age of the benzyl groups by hydrogenation using Pd/C, H2,
1:1) in a yield of 40%. Separation of 16 and 17 was possible and Et3N, the ratio was identical.
after acetylation of the anomeric position under standard
Next, the fluorinated glycopyranosyl phosphates 21–23
conditions. After separation, selective cleavage of the anom- were converted into the corresponding NDP sugars by reac-
eric acetyl function was achieved with hydrazinium acetate tion with 2. The target compounds were obtained in a mod-
in DMF (Scheme 4).[30]
erate yield of 52% [UDP-2-fluoro-2-deoxy-α-d-mannose
The phosphoramidite method was used to convert the (24)], a high yield of 72% [UDP-2-fluoro-2-deoxy-α/β-d-
fluorinated sugars into the α-phosphates (Scheme 5).[31] glucose (25)], and a very high yield of 91% [UDP-2-fluoro-
Compounds 15–17 were treated with dibenzyl N,N-diiso- 2-deoxy-α/β-d-galactose (26)] and still showed the same ra-
propylphosphoramidite and 4,5-dicyanoimidazole (DCI) to tios of α/β anomers after coupling (Figure 4).
give the phosphite intermediates, which were oxidized by
m-chloroperbenzoic acid to give benzyl-protected phos-
phates in yields of about 70%. Phosphorylation of the man-
nose compound yielded only the α-anomer due to the trans-
diaxial substituents at the C1 and C2 atoms. Unfortunately,
the obtained α/β mixtures of benzyl-protected glucose and
galactose phosphates were inseparable, even after several
Synthesis of L-Configured NDP Sugar 27
As an additional example of an l-configured sugar nu-
cleotide, UDP-β-l-galactose (27) was synthesized
(Scheme 6). Therefore, l-galactono-1,4-lactone (28) was re-
duced with NaBH4 in the presence of Amberlite and per-
acetylated in one pot (2 steps, 84%).[32] After bromination
of the anomeric position (96%), the bromide was converted
selectively into the benzyl-protected β-phosphate (66%
yield) by using dibenzyl phosphate and addition of
Ag2CO3. After hydrogenation (74%), using the conditions
as above, 1,2,3,4-tetra-O-acetyl-β-l-galactose phosphate
(29) was coupled with 2. Coupling and deprotection led to
the expected product in a yield of 63%.
Application of Nucleoside Analogues
As an example of a nucleoside analogue, we have already
published the synthesis of 3Ј-deoxy-2Ј,3Ј-didehydrothymid-
ine-α-d-glucose (d4TDP-α-d-glucose), which was obtained
in 78% yield.[24] To demonstrate that the method was not
restricted to certain nucleobases, five NDP glycopyranoses
were synthesized. In addition to 3Ј-deoxythymidine (30;
ddT), which was prepared by hydrogenation of d4T 31, 3Ј-
azido-3Ј-deoxythymidine (32; AZT), the fluorescent nu-
cleosides iso-2Ј,3Ј-dideoxyadenosine (33; iso-ddA), and
carbocyclic iso-2Ј,3Ј-dideoxyadenosine (34; iso-carba-ddA)
were used. These nucleoside analogues were prepared ac-
cording to known literature procedures (Figure 5).[33–36]
Scheme 5. Synthesis of fluorinated glycopyranosyl phosphates 21–
23.
Figure 4. Fluorinated target compounds 24–26.
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Synthesis of Nonnatural Nucleoside Diphosphate Sugars
Scheme 6. Synthesis of UDP-β-l-galactose (27) starting from l-galactono-1,4-lactone (28).
Figure 5. Nucleoside analogues 30 and 32–34.
All nucleoside analogues were converted into their corre-
sponding 5-nitro-cycloSal phosphate triesters by reaction
with 5-nitro-cycloSal-chlorophosphite followed by oxi-
dation and then treatment with anomerically pure glycopyr-
anosyl phosphates to give 35 (59%), 36 (40%), 37 (58%),
and 38 (56%). Moreover, d4TDP-α-d-mannose (39) was
also prepared in a yield of 70% (Scheme 7).
Scheme 7. Synthesis of NDP sugars 35–39 with nucleoside ana-
logues.
The two nucleoside analogues 31 and 32 are known anti-
viral drugs used as chemotherapeutic agents. For this
reason, NDP sugars with these nucleoside analogues were
tested for their antiviral activity in an HIV-1 and HIV-2
infected human T-lymphocyte cell line [wild-type CEM-
cells (CEM/O) as well as mutant CEM/thymidine kinase
deficient (CEM/TK-def) cells]. Although the compounds
disclosed herein showed some antiviral effect in the submi-
cromolar range (EC50: 0.5 μm for 39 and 0.04 μm for 38) in
the wild-type CEM cells, they were devoid of any antiviral
lated to the high polarity of the NDP sugars due to the
remaining charges at the pyrophosphate linkage, which pre-
vent membrane permeability.
Conclusions
We have proven that the cycloSal-approach can be ap-
effect in the mutant CEM/TK-def cells. This may be due to plied to a wide range of nucleosides and nucleoside ana-
extracellular enzymatic or chemical hydrolysis of the NDP logues. In addition, so far, we have not observed any restric-
sugars to give the parent nucleoside analogues, which are tion with respect to the configuration or modification of
able to penetrate the cell membranes, and, thus, cause an the sugar moieties. It seems that the method offers universal
antiviral effect. However, another reason may be that the access to a large number of very interesting sugar nucleo-
NDP sugars are unable to enter the cells in an intact form, tides and their analogues. Moreover, the example of 6
because if this were true then some activity should also be proved that post-synthetic modifications could also be
observed in the mutant CEM/TK-def cells. This may be re- achieved with NDP sugars.
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NMR (101 MHz, CDCl3): δ = 174.9 (2ϫ Cq-acetyl), 170.4 (2ϫ Cq-
acetyl), 164.6 (2ϫ C4base), 153.6 (2ϫ C2ar), 150.7 (2ϫ C2base),
141.6 (2ϫ C5ar), 139.2 (2ϫ C6base), 131.1 (2ϫ C1ar), 125.7 (2ϫ
C4ar), 121.5 (2ϫ C6ar), 119.5 (2ϫ C3ar), 103.4 (2ϫ C5base), 88.3
(2ϫ C1Ј), 80.4 (2ϫ C4Ј), 72.4 (2ϫ C2Ј), 69.5 (2ϫ C3Ј), 68.1 (2ϫ
C7ar), 67.4 (2ϫ C5Ј), 20.5 (2ϫ CH3-acetyl), 20.4 (2ϫ CH3-acetyl)
ppm. 31P NMR (162 MHz, CDCl3): δ = –10.32 (s), –10.69 (s) ppm.
Experimental Section
General: All experiments were conducted under rigorously dry con-
ditions and under nitrogen. Acetonitrile and diisopropylethylamine
(DIPEA) were dried by heating under reflux over calcium hydride
for several days, followed by distillation. Acetonitrile was stored
over activated molecular sieves. N,N-Dimethylformamide was pur-
chased from Fluka and was stored over molecular sieves. Ethyl
acetate and dichloromethane for extraction were distilled before
use. Analytical TLC was performed on Merck precoated alumin-
ium plates (60 F254) with a 0.2 mm layer of silica gel containing
a fluorescence indicator; compounds were visualized with a spray
reagent [4-methoxybenzaldehyde (0.5 mL), EtOH (9 mL), concd.
H2SO4 (0.5 mL), glacial AcOH (0.1 mL)] by heating with a drying
fan. NMR spectroscopic data were recorded with Bruker AMX
400 (400 MHz), Bruker DMX 500 (500 MHz) or Bruker AV 400
(400 MHz) spectrometers. All 1H and 13C NMR chemical shifts are
quoted in ppm and were calibrated by solvent signals if possible.
31P NMR chemical shifts are quoted in ppm by using H3PO4 as an
external reference. High-resolution mass spectra were obtained
with a VG Analytical VG/70-250F spectrometer (FAB, m-ni-
trobenzyl alcohol matrix). HR-ESI spectra were obtained with an
Agilent Technologies ESITOF 6224 spectrometer. Optical rotations
were measured with a P8000 polarimeter (A. Kruss Optonic
GmbH) at 589 nm. IR spectra were recorded with a ThermoNicolet
Avatar 370 FT-IR spectrometer.
IR (KBr): ν = 3434, 2926, 1751, 1701, 1627, 1561, 1486, 1377, 1248,
˜
1091, 1034, 940, 909, 816 cm–1. HRMS (ESI+): calcd. for
C20H21N3O13P+ [M + H+] 542.0807; found 542.0806.
5-Nitro-cycloSal-3Ј-deoxy-3Ј-dehydrothymidine
Monophosphate:
According to the general procedure, the reaction was conducted
with 31 (143 mg, 0.64 mmol), 5-nitro-cycloSal-chlorophosphite
(295 mg, 1.27 mmol), DIPEA (0.22 mL, 1.27 mmol), Oxone®
(1.55 mg, 2.52 mmol) and acetonitrile (20 mL) within 5 h (265 mg,
1
0.61 mmol, 95%). H NMR (400 MHz, [D6]DMSO): δ = 11.35 (s,
4
4
2 H, 2ϫ NH), 8.31 (dd, JHH = 2.9, JHH = 2.4 Hz, 2 H, 2ϫ 6ar-
3
4
H), 8.25 (dd, JHH = 9.1, JHH = 2.4 Hz, 2 H, 2ϫ 4ar-H), 7.38 (d,
3JHH = 9.1 Hz, 2 H, 2ϫ 3ar-H), 7.18–7.16 (m, 2 H, 2ϫ 6base-H),
6.81–6.76 (m, 2 H, 2ϫ 1Ј-H), 6.44–6.37 (m, 2 H, 2ϫ 2Ј-H), 6.05–
6.01 (m, 2 H, 2ϫ 3Ј-H), 5.69–5.60 (m, 2 H, 2ϫ 7ar-H), 5.55–5.48
(m, 2 H, 2ϫ 7ar-H), 4.97–4.96 (m, 2 H, 2ϫ 4Ј-H), 4.40–4.31 (m, 4
4
4
H, 2ϫ 5Ј-H), 1.68 (d, JHH = 1.1 Hz, 3 H, CH3), 1.65 (d, JHH
=
1.1 Hz, 3 H, CH3) ppm. 13C NMR (101 MHz, [D6]DMSO): δ =
163.6 (2ϫ C4base), 153.9 (2ϫ C5ar), 150.5 (2ϫ C2base), 143.4 (2ϫ
C2ar), 135.5 (2ϫ C6base), 132.5 (2ϫ C3Ј), 127.3 (3ϫ C2Ј), 125.4
(2ϫ C4ar), 122.4 (2ϫ C6ar), 119.8 (2ϫ C1ar), 119.4 (2ϫ C3ar),
109.5 (2ϫ C5base), 89.2 (2ϫ C1Ј), 83.9 (2ϫ C4Ј), 68.8 (2ϫ C5Ј),
67.5 (2ϫ C7ar), 11.8 (2ϫ CH3) ppm. 31P NMR (162 MHz, [D6]-
General Procedure for the Synthesis of 5-Nitro-Substituted cycloSal-
Nucleotides: Reactions were carried out under nitrogen. Acetyl-pro-
tected nucleosides, with unprotected 5Ј-hydroxy functionalities
(1.0 equiv.) were dissolved in anhydrous acetonitrile, and the solu-
tion was cooled to –20 °C. Diisopropyl(ethyl)amine (DIPEA,
2.0 equiv.) and 5-nitro-cycloSal-chlorophosphite dissolved in anhy-
drous acetonitrile were added. The reaction mixture was warmed
to room temperature and stirring continued until complete conver-
sion of the nucleoside (about 5 h). Subsequently, Oxone®
(4.0 equiv.) dissolved in water was added at –10 °C. Cooling was
removed, and stirring was continued for 15 min. The mixture was
then directly extracted with ethyl acetate and cold water (3 times),
the organic layer was dried with sodium sulfate, and the solvent
was removed under reduced pressure. The resulting products were
redissolved in dichloromethane. After filtration of the precipitate,
the solvent was again removed in vacuo. Lyophilization yielded the
products as colorless foams in a diastereomeric mixture. The tri-
esters obtained were of high purity and could be used as crude
materials without further purification.
DMSO): δ = –10.31 (s), –10.56 (s) ppm. IR (KBr): ν = 3082, 1685,
˜
1588, 1465, 1345, 1248, 1085, 1028, 994, 936 cm–1. HRMS (FAB):
calcd. for C17H17N3O9P+ [M + H+] 438.0697; found 438.0709.
5-Nitro-cycloSal-3Ј-deoxythymidine Monophosphate: According to
the general procedure, the reaction was conducted with 30 (180 mg,
0.80 mmol), 5-nitro-cycloSal-chlorophosphite (450 mg, 1.93 mmol),
DIPEA (0.28 mL, 1.55 mmol), Oxone® (2.13 mg, 3.46 mmol), and
acetonitrile (30 mL) within 5 h to give the product (282 mg,
1
0.64 mmol, 80%). H NMR (400 MHz, [D6]DMSO): δ = 11.29 (s,
2 H, 2ϫ NH), 8.31 (s, 2 H, 2ϫ 6ar-H), 8.27–8.23 (m, 2 H, 2ϫ 3ar-
H), 7.45–7.37 (m, 4 H, 2ϫ 6base-H, 2ϫ 4ar-H), 6.00–5.97 (m, 2 H,
2ϫ 1Ј-H), 5.72–5.62 (m, 2 H, 2ϫ 7ar-H) 5.57–5.52 (m, 2 H, 2ϫ
7ar-H), 4.47–4.29 (m, 4 H, 2ϫ 5Ј-H), 4.19–4.17 (m, 2 H, 2ϫ 4Ј-H),
2.29–2.24 (m, 2 H, 2ϫ 2Ј-H), 2.03–1.91 (m, 4 H, 2ϫ 2Ј-H, 2ϫ 3Ј-
H), 1.87–1.82 (m, 2 H, 2ϫ 3Ј-H), 1.75 (s, 3 H, CH3), 1.72 (s, 3 H,
CH3) ppm. 13C NMR (101 MHz, [D6]DMSO): δ = 164.2 (2ϫ
C4base), 154.5 (2ϫ C5ar), 150.9 (2ϫ C2base), 144.0 (2ϫ C2ar), 136.3
(2ϫ C6base), 127.8 (2ϫ C4ar), 126.0 (2ϫ C6ar), 123.1 (2ϫ C1ar),
120.2 (2ϫ C3ar), 109.5 (C5base), 85.4 (2ϫ C1Ј), 78.2 (2ϫ C4Ј), 69.9
(2ϫ C5Ј), 68.4 (2ϫ C7ar), 30.7 (2ϫ C2Ј), 25.7 (2ϫ C3Ј), 12.6 (2ϫ
CH3) ppm. 31P NMR (162 MHz, [D6]DMSO): δ = –10.57 (s),
5-Nitro-cycloSal-2Ј,3Ј-O-acetyluridine Monophosphate (2): Accord-
ing to the general procedure, the reaction was conducted with 2Ј,3Ј-
OAc-uridine (300 mg, 0.91 mmol), 5-nitro-cycloSal-chlorophosphite
(419 mg, 1.80 mmol), DIPEA (0.32 mL, 1.83 mmol), Oxone®
(2.25 g, 3.66 mmol), and acetonitrile (25 mL) within 6 h to give 2
1
(460 mg, 0.85 mmol, 93%). H NMR (400 MHz, CDCl3): δ = 1.11
–10.95 (s) ppm. IR (KBr): ν = 3448, 3075, 1686, 1589, 1528, 1483,
˜
(s, 1 H, 1-H), 1.11 (s, 1 H, 1-H), 1.11 (s, 1 H, 1-H), 8.46 (s, 2 H,
2ϫ NH), 8.26–8.25 (m, 1 H, 4ar-H), 8.24–8.23 (m, 1 H, 4ar-H),
1346, 1256, 1090, 1034, 938 cm–1. HRMS (FAB): calcd. for
C17H19N3O9P+ [M + H+] 440.0853; found 440.0855.
3
8.09–8.08 (m, 2 H, 2ϫ 6ar-H), 7.38 (d, JHH = 8.2 Hz, 1 H, 6base
-
H), 7.36 (d, JHH = 8.2 Hz, 1 H, 6base-H), 7.24 (d, JHH = 9.1 Hz, 5-Nitro-cycloSal-2Ј,3Ј-dideoxy-N4-iso-adenosine
Monophosphate:
3
3
3
3
1 H, 3ar-H), 7.23 (d, JHH = 9.1 Hz, 1 H, 3ar-H), 5.94 (d, JHH
=
According to the general procedure, the reaction was conducted
3
6.8 Hz, 1 H, 1Ј-H), 5.93 (d, JHH = 6.8 Hz, 1 H, 1Ј-H), 5.74 (d, with 33 (40 mg, 0.17 mmol), 5-nitro-cycloSal-chlorophosphite
3JHH = 8.2 Hz, 1 H, 5base-H), 5.72 (d, 3JHH = 8.2 Hz, 1 H, 5base-H), (80 mg, 0.34 mmol), DIPEA (0.06 mL, 0.32 mmol), Oxone®
5.53–5.47 (m, 4 H, 2ϫ 7ar-H), 5.39 (dd, 3JHH = 6.0, 3JHH = 4.7 Hz,
(420 mg, 0.68 mmol), and acetonitrile (15 mL) within 5 h to give
1 H, 3Ј-H), 5.32 (dd, JHH = 6.0, JHH = 4.7 Hz, 1 H, 3Ј-H), 5.30– the product (25 mg, 0.06 mmol, 33%). 1H NMR (400 MHz, [D6]
3
3
5.26 (m, 2 H, 2ϫ 2Ј-H), 4.62–4.54 (m, 2 H, 5Ј-H), 4.52–4.42 (m, 2 DMSO): δ = 8.69 (s, 2 H, 2ϫ 6base-H), 8.53 (s, 2 H, 2ϫ 8base-H),
H, 5Ј-H), 4.35–4.30 (m, 2 H, 2ϫ 4Ј-H), 2.13 (s, 3 H, CH3-acetyl),
8.22–8.20 (m, 2 H, 2ϫ 3ar-H), 7.34–7.30 (m, 2 H, 2ϫ 4ar-H), 7.23–
3
2.11 (s, 3 H, CH3-acetyl), 2.09 (s, 6 H, 2ϫ CH3-acetyl) ppm. 13C 7.21 (m, 2 H, 2ϫ 6ar-H), 6.53 (s, 4 H, 2ϫ NH2), 6.15 (dd, JHH
=
6310
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© 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2011, 6304–6313

Synthesis of Nonnatural Nucleoside Diphosphate Sugars
4
6.6, JHH = 3.8 Hz, 2 H, 2ϫ 1Ј-H), 5.59–5.32 (m, 4 H, 2ϫ 7ar-H),
times). After lyophilization of the aqueous phase, acetyl groups
were cleaved by a mixture of methanol, water, and triethylamine
4.13–4.08 (m, 2 H, 2ϫ 4Ј-H), 3.45–3.35 (m, 4 H, 2ϫ 5Ј-H), 2.36–
2.26 (m, 4 H, 2ϫ 2Ј-H), 2.12–2.02 (m, 4 H, 2ϫ 3Ј-H) ppm. 13C (7:3:1) at room temperature overnight. After a second lyophiliza-
NMR (101 MHz, [D6]DMSO): δ = 160.2 (2ϫ C2base), 152.6 (2ϫ
C4base), 149.8 (2ϫ C6base), 148.6 (2ϫ C2ar), 140.8 (2ϫ C8base), an RP-18 column using water as the solvent. NDP pyranoses were
140.3 (2ϫ C5ar), 131.4 (2ϫ C1ar), 130.5 (2ϫ C4ar), 126.8 (2ϫ detected by TLC using 2-propanol and 1 n ammonium acetate
C5base), 124.6 (2ϫ C3ar), 117.2 (2ϫ C6ar), 83.7 (2ϫ C1Ј), 81.9 (2ϫ solution (2:1, v/v) as the mobile phase. Usually two rounds of
tion, the target compounds were purified by chromatography on
C4Ј), 68.6 (2ϫ C7ar), 58.6 (2ϫ C5Ј), 31.7 (2ϫ C2Ј), 25.8 (2ϫ C3Ј)
chromatography were sufficient to isolate the desired products in
high purity.
ppm. 31P NMR (162 MHz, [D6]DMSO): δ = –10.90 (s), –11.05 (s)
ppm. IR (KBr): ν = 2929, 2104, 1660, 1616, 1589, 1522, 1429, 1337,
˜
UDP-α-D-sulfoquinovose 4: According to the general procedure, the
1252, 1035 cm–1. HRMS (FAB): calcd. for C17H18N6O7P+ [M +
H+] 449.0969; found 449.0988.
reaction was conducted with 3 (215 mg, 0.43 mmol), 2, (92 mg,
0.17 mmol), and N,N-dimethylformamide (5 mL) within 24 h to
give 4 (76.4 mg, 0.08 mmol, 48%). 1H NMR (400 MHz, D2O): δ =
5-Nitro-cycloSal-2Ј,3Ј-dideoxy-N4-iso-carbaadenosine Monophos-
phate: According to the general procedure, the reaction was con-
ducted with 34 (148 mg, 0.64 mmol), 5-nitro-cycloSal-chlorophos-
phite (295 mg, 1.27 mmol), DIPEA (0.22 mL, 1.26 mmol), Oxone®
(1.55 g, 2.52 mmol), and acetonitrile (20 mL) within 5 h to give the
product (165 mg, 0.37 mmol, 58%). 1H NMR (400 MHz, [D6]-
3
7.99 (d, JHH = 8.1 Hz, 1 H, 6base-H), 6.03–6.02 (m, 2 H, 5base-H,
3
4
1Ј-H), 5.64 (dd, JHP = 8.1, JHH = 3.1 Hz, 1 H, 1-H), 4.44–4.42
(m, 2 H, 2Ј-H, 3Ј-H), 4.33–4.31 (m, 1 H, 5Ј-H), 4.27–4.25 (m, 2 H,
3
4Ј-H, 5Ј-H), 3.81 (dd, 3JHH = 9.5, JHH = 9.5 Hz, 1 H, 5-H), 3.61–
3
4
3.55 (m, 2 H, 2-H, 4-H), 3.43 (dd, JHH = 15.0, JHH = 3.1 Hz, 1
DMSO): δ = 8.57 (s, 2 H, 2ϫ 6base-H), 8.16 (s, 2 H, 2ϫ 8base-H), H, 3-H), 3.30–3.18 (m, 2 H, 6-H) ppm. 13C NMR (101 MHz, D2O):
3
3
2
7.25 (d, JHH = 2.9 Hz, 2 H, 4ar-H), 7.23 (d, JHH = 2.9 Hz, 1 H,
3ar-H), 7.06–7.04 (m, 2 H, 2ϫ 6ar-H), 6.53 (s, 4 H, 2ϫ NH2), 5.70–
δ = 166.3 (C4base), 151.9 (C2base), 102.8 (C5base), 95.7 (d, JCP
=
3
6.4 Hz, C1), 88.4 (C1Ј), 83.4 (d, JCP = 9.1 Hz, C2), 73.8 (C6base),
3
5.58 (m, 4 H, 2ϫ 7ar-H), 4.74–4.71 (m, 2 H, 2ϫ 1Ј-H), 4.29–4.21 72.7 (C2Ј), 71.8 (C4), 71.7 (d, JCP = 7.6 Hz, C4Ј), 69.8 (C5), 69.4
(m, 4 H, 2ϫ 6Ј-H), 2.43–2.39 (m, 2 H, 2ϫ 4Ј-H), 2.31–2.24 (m, 4 (C3Ј), 65.1 (d, JCP = 5.7 Hz, C5Ј), 51.9 (C3), 50.5 (C6) ppm. 31P
2
H, 2ϫ 5Ј-H), 2.14–1.99 (m, 4 H, 2ϫ 2Ј-H), 1.75–1.64 (m, 4 H, 2ϫ NMR (162 MHz, D2O): δ = –10.99 (d, JPP = 20.7 Hz, Pα), –12.87
3Ј-H) ppm. 13C NMR (101 MHz, [D6]DMSO): δ = 160.0 (2ϫ (d, JPP = 20.7 Hz, P ) ppm. IR (KBr): ν = 2933, 1677, 1453, 1034,
˜
β
C2base), 152.9 (2ϫ C4base), 149.3 (2ϫ C6base), 143.4 (2ϫ C2ar), 490 cm–1. HRMS (ESI–): calcd. for C15H23N2O19P2S– [M + 2H+]
139.6 (2ϫ C8base), 138.3 (2ϫ C5ar), 129.9 (2ϫ C3ar),127.2 (2ϫ 629.0096; found 629.0098. [α]2D4 = +11.8 (c = 0.1, H2O).
C5base), 124.8 (2ϫ C4ar), 124.3 (2ϫ C6ar), 121.8 (2ϫ C1ar), 76.8
UDP-6-sulfate-α-D-glucose 6: First, compound 5 was synthesized
(2ϫ C6Ј), 73.8 (2ϫ C7ar), 55.5 (2ϫ C1Ј), 38.9 (2ϫ C4Ј), 32.3 (2ϫ
according to the general procedure. Then, compound 5 (30 mg,
0.05 mmol) was dissolved in absolute pyridine (3 mL), and the sul-
fur trioxide–pyridine complex (13 mg, 0.08 mmol) was added. After
stirring at room temperature for 22 h, compound 6 was formed in
a quantitative yield (34 mg, 0.05 mmol). 1H NMR (400 MHz,
C5Ј), 31.1 (2ϫ 2Ј-H), 26.6 (2ϫ 3Ј-H) ppm. 31P NMR (162 MHz,
[D ]DMSO): δ = –10.84 (s), –10.86 (s) ppm. IR (KBr): ν = 3082,
˜
6
1660, 1524, 1344, 1296, 1249, 1025, 935, 904 cm–1. HRMS (FAB):
calcd. for C18H20N6O6P+ [M + H+] 447.1176; found 447.1190.
3
5-Nitro-cycloSal-3Ј-deoxy-3Ј-azidothymidine Monophosphate: Ac-
cording to the general procedure, the reaction was conducted with
32 (190 mg, 0.70 mmol), 5-nitro-cycloSal-chlorophosphite (330 mg,
1.41 mmol), DIPEA (0.25 mL, 1.40 mmol), Oxone® (1.75 g,
2.85 mmol), and acetonitrile (15 mL) within 5 h to give the product
D2O): δ = 8.01 (d, JHH = 8.2 Hz, 1 H, 6base-H), 6.01–5.99 (m, 2
H, 5base-H, 1Ј-H), 5.72–5.70 (m, 1 H, 1-H), 4.42–4.40 (m, 2 H, 2Ј-
H, 3Ј-H), 4.35–4.33 (m, 1 H, 5Ј-H), 4.30–4.28 (m, 2 H, 4Ј-H, 5Ј-
H), 3.79–3.76 (m, 3 H, 2-H, 4-H, 5-H), 3.31–3.19 (m, 3 H, 3-H, 6-
H) ppm. 13C NMR (101 MHz, D2O): δ = 161.5 (C4base), 153.7
(157 mg, 0.33 mmol, 47%). 1H NMR (400 MHz, CDCl3): δ = 8.26
(C2base), 102.3 (C5base), 96.8 (d, 2JCP = 6.7 Hz, C1), 89.6 (C1Ј), 84.7
3
3
(d, JHH = 8.5 Hz, 2 H, 2ϫ 4ar-H), 8.08 (s, 2 H, 2ϫ 6base-H),7.24– (d, JCP = 9.6 Hz, C2), 75.8 (C6base), 72.9 (C2Ј), 72.8 (C4), 71.6 (d,
2
7.21 (m, 4 H, 2ϫ 3ar-H, 2ϫ 6ar-H), 6.09–6.07 (m, 2 H, 2ϫ 1Ј-H), 3JCP = 7.3 Hz, C4Ј), 70.9 (C5), 69.1 (C3Ј), 67.3 (d, JCP = 6.1 Hz,
5.52–5.45 (m, 4 H, 2ϫ 7ar-H), 4.52–4.32 (m, 6 H, 2ϫ 4Ј-H, 2ϫ 5Ј-
H), 4.06–4.02 (m, 2 H, 2ϫ 3Ј-H), 2.48–2-42 (m, 4 H, 2ϫ 2Ј-H),
1.25 (s, 6 H, 2ϫ CH3) ppm. 13C NMR (101 MHz, CDCl3): δ =
C5Ј), 53.1 (C3), 60.4 (C6) ppm. 31P NMR (162 MHz, D2O): δ =
–11.24 (d, JPP = 19.9 Hz, Pα), –12.93 (d, JPP = 19.9 Hz, Pβ) ppm.
IR (KBr): ν = 2933, 1677, 1442, 1391, 1034, 613, 450 cm–1. HRMS
˜
166.3 (2ϫ C4base), 158.8 (2ϫ C2ar), 153.3 (2ϫ C2base), 141.3 (2ϫ (ESI–): calcd. for C15H23N2O20P2S– [M + 2H+] 645.0046; found
C5ar), 135.5 (2ϫ C4ar), 130.4 (2ϫ C1ar), 126.8 (2ϫ C3ar), 121.6
(2ϫ C6base), 119.4 (2ϫ C6ar), 103.4 (2ϫ C5base), 85.2 (2ϫ C1Ј),
81.9 (2ϫ C3Ј), 73.7 (2ϫ C2Ј), 68.1 (2ϫ C4Ј), 67.8 (2ϫ C7ar), 59.9
(2ϫ C5Ј), 29.7 (2ϫ CH3) ppm. 31P NMR (162 MHz, [D6]DMSO):
645.0051. [α]2D4 = –1.9 (c = 0.1, H2O).
UDP-2-fluoro-2-deoxy-α/β- -galactose 26: According to the general
D
procedure, the reaction was conducted with 21 (140 mg,
0.25 mmol), 2 (95.2 mg, 0.18 mmol), and N,N-dimethylformamide
(4 mL) within 24 h to give 26 (90.6 mg, 0.16 mmol, 91%) in an
anomeric ratio of α/β = 1:0.4. Characterization data for the α com-
δ = –10.13 (s), –10.29 (s) ppm. IR (KBr): ν = 3044, 2105, 1677,
˜
1460, 1336, 1253, 1071, 1033, 907, 846 cm–1. HRMS (FAB): calcd.
for C17H18N6O9P+ [M + H+] 481.0867; found 481.0869.
1
3
pound are given. H NMR (400 MHz, D2O): δ = 7.96 (d, JHH
=
General Procedure for the Synthesis of NDP Glycopyranoses: Reac-
tions were carried out under nitrogen, and dry conditions were
strictly adhered to. The corresponding glycopyranosyl phosphates
(2.0 equiv.) were dissolved in anhydrous N,N-dimethylformamide.
After storage over activated molecular sieves for 1 h, the corre-
sponding 5-nitro-cycloSal-nucleotides (1.0 equiv.) dissolved in an-
hydrous N,N-dimethylformamide were added within 10 min. The
reaction mixture was stirred until complete conversion of the tri-
ester (18–26 h). The solvent was removed under reduced pressure
followed by extraction with ethyl acetate and water (up to 10
8.1 Hz, 1 H, 6base-H), 6.02–5.97 (m, 2 H, 5base-H, 1Ј-H), 5.84 (dd,
3
3JHF = 7.5, JHH = 3.7 Hz, 1 H, 1-H), 4.67–4.63 (m, 1 H, 3Ј-H),
4.40–4.38 (m, 2 H, 2Ј-H, 4Ј-H), 4.30–4.27 (m, 1 H, 2-H), 4.25–4.19
(m, 3 H, 5-H, 5Ј-H), 4.11–4.10 (m, 1 H, 3-H), 4.02–3.97 (m, 1 H,
6-H), 3.84–3.74 (m, 2 H, 4-H, 6-H) ppm. 13C NMR (101 MHz,
D2O): δ = 166.0 (C4base), 161.4 (C2base), 141.5 (C6base), 102.7
(C5base), 92.8 (dd, 2JCF = 22.3, 2JCP = 5.4 Hz, C1), 88.4 (C1Ј), 87.2
3
(C3Ј), 83.2 (d, JCP = 9.3 Hz, C4Ј), 76.0 (C4), 73.1 (C2Ј), 71.0 (d,
2
3JCP = 18.0 Hz, C2), 69.7 (C3), 69.2 (C5), 64.9 (d, JCP = 5.4 Hz,
C5Ј), 60.8 (C6) ppm. 31P NMR (162 MHz, D2O): δ = –11.57 (d,
Eur. J. Org. Chem. 2011, 6304–6313
© 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
6311

S. Wolf, R. M. Berrio, C. Meier
FULL PAPER
JPP = 19.7 Hz, Pα), –13.30 (d, JPP = 19.7 Hz, Pβ) ppm. 19F NMR
ppm. IR (KBr): ν = 3213, 1680, 1462, 1231, 1035, 939, 815, 488,
˜
2
3
3
–
(200 MHz, D2O): δ = –208.0 (ddd, JFH = 57.0, JFH = 16.0, JFH 414 cm–1. HRMS (ESI–): calcd. for C15H23N2O17P2 [M + H+]
= 5.0 Hz) ppm. IR (KBr): ν = 3300, 1672, 1232, 1052, 982, 927, 565.0477; found 565.0475. [α]2D4 = +3.8 (c = 0.1, H2O).
˜
–
859 cm–1. HRMS (ESI–): calcd. for C15H22FN2O16P2 [M + H+]
ddT-α-D-glucose 35: According to the general procedure, the reac-
567.0434; found 567.0440. [α]2D4 = –1.7 (c = 0.1, H2O).
tion was conducted with α-d-glucopyranosyl phosphate (94 mg,
0.36 mmol), 5-nitro-cycloSal-3Ј-deoxythymidine monophosphate,
(80 mg, 0.18 mmol), and N,N-dimethylformamide (5 mL) within
18 h to give 35 (58.4 mg, 0.11 mmol, 59%). 1H NMR (400 MHz,
UDP-2-fluoro-2-deoxy-α/β-D-glucose 25: According to the general
procedure, the reaction was conducted with 22 (132 mg,
0.34 mmol), 2 (92.5 mg, 0.17 mmol) and N,N-dimethylformamide
(4 mL) within 24 h to give 25 (69.6 mg, 0.12 mmol, 72%) in an D2O): δ = 7.78 (d, JHH = 1.2 Hz, 1 H, 6base-H), 6.14 (dd, JHH
anomeric ratio of α/β = 1:0.4. Characterization data for the α com-
4
3
=
7.0, 4JHH = 4.0 Hz 1 H, 1Ј-H), 5.61 (dd, 3JHP = 7.3, 3JHH = 3.3 Hz,
1 H, 1-H), 4.39–4.38 (m, 1 H, 5Ј-H), 4.26 (ddd, JHH = 11.3, JHH
1
3
2
3
pound are given. H NMR (400 MHz, D2O): δ = 7.95 (d, JHH
=
3
4
2
3
8.0 Hz, 1 H, 6base-H), 6.03–6.01 (m, 1 H, 5base-H), 5.97 (dd, JHH = 5.6, JHH = 2.8 Hz, 1 H, 6-H), 4.12 (ddd, JHH = 11.3, JHH
=
4
3
3
= 8.0, JHH = 1.6 Hz, 1 H, 1Ј-H), 5.79 (dd, JHF = 7.5, JHH
=
6.2, 4JHH = 4.7 Hz, 1 H, 6-H), 3.94–3.85 (m, 2 H, 4-H, 5Ј-H), 3.82–
3.76 (m, 2 H, 3-H, 5-H), 3.53 (ddd, JHH = 9.8, JHH = 3.3, JHH
= 3.2 Hz, 1 H, 2-H), 3.49–3.44 (m, 1 H, 4Ј-H), 2.47–2.40 (m, 1 H,
3.6 Hz, 1 H, 1-H), 4.38 (d, 3JHH = 3.9 Hz, 1 H, 3Ј-H), 4.32 (s, 2 H,
3
3
4
3
2Ј-H, 4Ј-H), 4.24 (s, 1 H, 2-H), 3.99 (d, JHH = 7.9 Hz, 1 H, 3-H),
3.69 (dd, JHH = 9.5, JHH = 7.9 Hz, 3 H, 5-H, 5Ј-H), 3.50–3.46 2Ј-H), 2.19–2.12 (m, 2 H, 3Ј-H), 2.08–2.03 (m, 1 H, 2Ј-H), 1.95 (s,
3
3
2
3
(m, 1 H, 6-H), 3.38 (dd, JHH = 18.4, JHH = 9.5 Hz, 2 H, 4-H, 6-
3 H, CH3) ppm. 13C NMR (101 MHz, D2O): δ = 168.9 (C4base),
H) ppm. 13C NMR (101 MHz, D2O): δ = 170.6 (C4base), 162.4 150.8 (C2base), 137.2 (C6base),95.5 (d, JCP = 6.8 Hz, C1), 88.1
2
2
2
3
(C2base), 141.4 (C5base), 102.8 (C6base), 92.6 (dd, JCF = 20.9, JCP (C1Ј), 80.2 (d, JCP = 8.6 Hz, C2), 111.3 (C5base), 72.9 (C4), 72.8
3
= 5.0 Hz, C1), 88.4 (C1Ј),83.2 (d, JCP = 9.3 Hz, C4Ј), 72.7 (C2Ј),
64.9 (d, 2JCP = 5.8 Hz, C5Ј), 69.8 (d, 3JCP = 6.0 Hz, C2),76.6 (C3Ј),
73.8 (C4), 70.9 (C3), 68.7 (C5), 60.1 (C6) ppm. 31P NMR
(162 MHz, D2O): δ = –11.59 (d, JPP = 19.8 Hz, Pα), –13.42 (d, JPP
= 19.8 Hz, Pβ) ppm. 19F NMR (200 MHz, D2O): δ = –200.0 (dd,
(C5), 71.7 (d, 3JCP = 8.5 Hz, C4Ј), 69.3 (C3), 66.9 (d, 2JCP = 5.8 Hz,
C5Ј), 60.4 (C6), 31.1 (C2Ј), 25.0 (C3Ј), 11.7 (CH3) ppm. 31P NMR
(162 MHz, D2O): δ = –11.09 (d, JPP = 21.1 Hz, Pα), –12.91 (d, JPP
= 21.1 Hz, P ) ppm. IR (KBr): ν = 3246, 2107, 1681, 1475, 1232,
˜
β
1081, 1046, 1002, 918, 505, 418 cm–1. HRMS (ESI–): calcd. for
–
2JFH = 49.2, 3JFH = 12.8 Hz) ppm. IR (KBr): ν = 3250, 1676, 1244,
C16H25N2O15P2 [M + H+] 547.0736; found 547.0734. [α]2D4 = +4.6
˜
1050, 972, 925, 814 cm–1. HRMS (ESI–): calcd. for
C15H22FN2O16P2– [M + H+] 567.0434; found 567.0438. [α]2D4 = +7.5
(c = 0.1, H2O).
(c = 0.1, H2O).
isoddA-β-D-glucose 36: According to the general procedure, the re-
action was conducted with 2,3,4,6-tetra-OAc-β-d-glucopyranosyl
phosphate (38 mg, 0.09 mmol), 5-nitro-cycloSal-2Ј,3Ј-dideoxy-N4-
iso-adenosine monophosphate, (20 mg, 0.04 mmol), and N,N-di-
methylformamide (3 mL) within 20 h to give 36 (10.1 mg,
UDP-2-fluoro-2-deoxy-α-D-mannose 24: According to the general
procedure, the reaction was conducted with 23 (73.7 mg,
0.19 mmol), 2 (51.5 mg, 0.09 mmol) and N,N-dimethylformamide
(4 mL) within 24 h to give 24 (38.1 mg, 0.05 mmol, 52%) as a pure 0.02 mmol, 40%). 1H NMR (400 MHz, D2O): δ = 8.66 (s, 1 H,
1
3
α-anomer. H NMR (400 MHz, D2O): δ = 8.00 (d, JHH = 8.0 Hz,
6
base-H), 8.42 (s, 1 H, 8base-H), 6.33 (dd, 3JHH = 6.6, 4JHH = 4.0 Hz,
3
3
3
1 H, 6base-H), 6.02–60 (m, 2 H, 5base-H, 1Ј-H), 5.65 (d, JHH
=
1 H, 1Ј-H), 5.00 (dd, JHP = 8.0, JHH = 8.0 Hz, 1 H, 1-H), 4.24–
4.5 Hz, 1 H, 1-H), 4.95 (s, 1 H, 3Ј-H), 4.41 –4.40 (s, 2 H, 2Ј-H, 4Ј- 4.19 (m, 1 H, 5-H), 4.13–4.07 (m, 1 H, 6-H), 3.89 (dd, 2JHH = 12.4,
2
3
H), 4.32–4.23 (m, 3 H, 5-H, 5Ј-H), 4.03 (dd, JHF = 30.8, JHH
9.8 Hz, 1 H, 2-H),3.93–3.91 (m, 2 H, 3-H, 6-H), 3.86–3.76 (m, 2 3.40–3.35 (m, 3 H, 5Ј-H, 2-H), 2.62–2.54 (m, 2 H, 2Ј-H), 2.30–2.19
H, 4-H, 6-H) ppm. 13C NMR (101 MHz, D2O): δ = 166.2 (C4base), (m, 2 H, 3Ј-H) ppm. 13C NMR (101 MHz, D2O): δ = 160.4
=
3JHH = 2.2 Hz, 1 H, 6-H), 3.56–3.49 (m, 3 H, 4Ј-H, 3-H, 4-H),
151.8 (C2base), 141.6 (C6base), 102.7 (C5base), 93.5 (dd, 2JCF = 30.8, (C2base), 152.9 (C4base), 149.7 (C6base), 140.3 (C8base), 126.1
2JCP = 9.8 Hz, C1), 90.3 (d, JCP = 10.3 Hz, C2), 88.9 (C3Ј), 88.4
(C5base), 96.5 (d, JCP = 6.7 Hz, C1), 83.9 (C1Ј), 73.6 (C5), 72.4 (d,
3
2
(C1Ј), 83.2 (d, 3JCP = 9.1 Hz, C4Ј), 73.8 (C4), 73.6 (C2Ј), 69.6 (C3),
3JCP = 6.1 Hz, C2), 72.3 (C3), 69.8 (d, JCP = 2.9 Hz, C4Ј), 68.6
3
69.2 (C5), 64.9 (d, JCP = 5.5 Hz, C5Ј), 60.3 (C6) ppm. 31P NMR (C4), 65.1 (d, 2JCP = 3.4 Hz, C5Ј), 60.2 (C6), 31.6 (C2Ј), 25.1 (C3Ј)
2
(162 MHz, D2O): δ = –11.49 (d, JPP = 19.8 Hz, Pα), –14.03 (d, JPP
ppm. 31P NMR (162 MHz, D2O): δ = –11.12 (d, JPP = 19.9 Hz,
= 19.8 Hz, Pβ) ppm. 19F NMR (200 MHz, D2O): δ = –204.7 (ddd,
Pα), –12.95 (d, JPP = 19.9 Hz, P ) ppm. IR (KBr): ν = 3516, 2924,
˜
β
2JFH = 49.4, 3JFH = 24.7, 3JFH = 5.9 Hz) ppm. IR (KBr): ν = 2982,
2853, 1620, 1581, 1429, 1052, 794, 640, 452, 414 cm–1. HRMS
˜
1682, 1065, 939, 801 cm–1. HRMS (ESI–): calcd. for
(ESI–): calcd. for C16H24N5O13P2 [M + H+] 556.0851 found;
–
2–
C15H21FN2O16P2 [M] 566.0361; found 566.1430. [α]2D4 = +2.0 (c
556.0870. [α]2D4 = –5.8 (c = 0.1, H2O).
= 0.1, H2O).
iso-carba-ddA-β-D-glucose 37: According to the general procedure,
UDP-β-
L
-galactose 27: According to the general procedure, the re-
the reaction was conducted with 2,3,4,6-tetra-OAc-β-d-glucopyr-
anosyl phosphate (275 mg, 0.52 mmol), 5-nitro-cycloSal-2Ј,3Ј-dide-
oxy-N4-iso-adenosine monophosphate, (116 mg, 0.26 mmol) and
N,N-dimethylformamide (5 mL) within 20 h to give 37 (81.5 mg,
0.15 mmol, 58%). 1H NMR (400 MHz, D2O): δ = 8.66 (s, 1 H,
action was conducted with 1.7 instead of 2.0 equiv. 29 (120 mg,
0.22 mmol), 2 (70 mg, 0.13 mmol), and N,N-dimethylformamide
(4 mL) within 22 h to give 27 (76.5 mg, 0.08 mmol, 63%). 1H NMR
3
(400 MHz, D2O): δ = 8.00 (d, JHH = 8.1 Hz, 1 H, 6base-H), 6.04–
6.02 (m, 2 H, 5base-H, 1Ј-H), 5.00 (dd, JHP = 7.7, JHH = 7.7 Hz, 6base-H), 8.35 (s, 1 H, 8base-H), 4.85 (dd, 3JHH = 8.8, 4JHH = 7.6 Hz,
3
3
3
3
1 H, 1-H), 4.43–4.42 (m, 2 H, 2Ј-H, 3Ј-H), 4.33–4.26 (m, 3 H, 4Ј-
H, 5Ј-H), 3.97–3.96 (m, 1 H, 5-H), 3.89–3.63 (m, 5 H, 2-H, 3-H,
1 H, 1Ј-H), 4.80 (dd, JHP = 8.5, JHH = 7.1 Hz, 1 H, 1-H), 3.97–
3.96 (m, 2 H, 6Ј-H), 3.84 (m, 1 H, 5-H), 3.79–3.74 (m, 1 H, 6-H),
4-H, 6-H) ppm. 13C NMR (101 MHz, D2O): δ = 166.2 (C4base), 3.58–3.56 (m, 4 H, 2-H, 3-H, 4-H, 6-H), 2.50–2.44 (m, 2 H, 4Ј-H,
151.9 (C2base), 141.7 (C6base), 102.8 (C5base), 98.6 (d, 2JCP = 6.1 Hz, 5Ј-H), 2.32–2.27 (m, 1 H, 5Ј-H), 2.00–1.90 (m, 2 H, 2Ј-H), 1.78–
C1), 88.4 (C1Ј), 83.4 (d, 3JCP = 9.1 Hz, C4Ј), 75.9 (C2Ј), 73.8 (C3Ј),
1.73 (m, 2 H, 3Ј-H) ppm. 13C NMR (101 MHz, D2O): δ = 163.4
3
72.4 (C4), 71.4 (d, JCP = 8.5 Hz, C2), 69.8 (C3), 68.7 (C5), 65.1 (C2base), 150.9 (C4base), 149.2 (C6base), 143.3 (C8base), 129.1
(d, 2JCP = 5.5 Hz, C5Ј), 61.3 (C6) ppm. 31P NMR (162 MHz, D2O): (C5base), 98.5 (d, JCP = 6.7 Hz, C1), 75.9 (C5), 72.4 (d, JCP
=
2
3
δ = –11.16 (d, JPP = 19.6 Hz, Pα), –12.74 (d, JPP = 19.6 Hz, Pβ) 6.1 Hz, C2), 72.3 (C3), 69.9 (C6Ј), 68.6 (C4), 61.2 (C6), 54.9 (C1Ј),
6312
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© 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2011, 6304–6313

Synthesis of Nonnatural Nucleoside Diphosphate Sugars
3
2
[2] N. K. Kochetkov, V. N. Shibaev, Adv. Carbohydr. Chem. Bio-
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37.8 (d, JCP = 2.9 Hz, C4Ј), 35.1 (d, JCP = 3.4 Hz, C5Ј), 31.3
(C2Ј), 26.1 (C3Ј) ppm. 31P NMR (162 MHz, D2O): δ = –10.73 (d,
JPP = 19.6 Hz, Pα), –12.98 (d, JPP = 19.6 Hz, Pβ) ppm. IR (KBr):
ν = 3257, 1613, 1509, 1454, 1421, 1224, 1047, 943, 789, 451 cm–1.
˜
–
HRMS (ESI–): calcd. for C17H26N5O12P2 [M + H+] 554.1059;
found 554.1071. [α]2D4 = –2.3 (c = 0.1, H2O).
AZT-α-D-glucose 38: According to the general procedure, the reac-
tion was conducted with α-d-glucopyranosyl phosphate (87 mg,
0.33 mmol), 5-nitro-cycloSal-3Ј-deoxy-3Ј-azidothymidine mono-
phosphate, (80 mg, 0.17 mmol), and N,N-dimethylformamide
(5 mL) within 18 h to give 38 (58.4 mg, 0.11 mmol, 56%). 1H NMR
3
(400 MHz, D2O): δ = 7.74 (s, 1 H, 6base-H), 6.28 (dd, JHH = 6.3,
3JHH = 6.3 Hz, 1 H, 1Ј-H), 5.61 (dd, JHH = 6.7, JHH = 3.3 Hz, 1
3
4
3
3
H, 1-H), 4.57 (dd, JHH = 8.8, JHH = 5.3 Hz, 1 H, 3Ј-H), 4.21–
4.20 (m, 3 H, 4Ј-H, 5Ј-H, 6-H), 4.92–4.85 (m, 2 H, 5Ј-H, 6-H),
3.80–3.76 (m, 2 H, 3-H, 5-H), 3.55–3.51 (m, 1 H, 2-H), 3.46 (dd,
4JHH = 9.6, JHH = 9.6 Hz, 1 H, 4-H), 2.50 (dd, JHH = 6.3, JHH
= 6.3 Hz, 2 H, 2Ј-H), 1.94 (s, 3 H, CH3) ppm. 13C NMR (101 MHz,
D2O): δ = 166.6 (C4base), 151.7 (C2base), 137.3 (C6base), 111.8
4
3
3
[9] D. Roche, J. Greiner, A.-M. Aubertin, P. Vierling, Bioconjugate
Chem. 2006, 17, 1568–1581.
[10] S. Roseman, J. J. Distler, J. G. Moffatt, H. G. Khorana, J. Am.
Chem. Soc. 1961, 83, 649–659.
2
(C5base), 95.6 (d, JCP = 6.5 Hz, C1), 85.7 (C1Ј), 85.0 (C3Ј), 83.0
[11] J. G. Moffatt, H. G. Khorana, J. Am. Chem. Soc. 1958, 80,
(d, 3JCP = 9.0 Hz, C2), 79.2 (C2Ј), 74.0 (C4), 72.9 (d, 3JCP = 6.8 Hz,
3756–3761.
2
C4Ј), 71.7 (d, JCP = 8.1 Hz, C5Ј), 60.4 (C6), 69.4 (C3), 69.3 (C5),
[12] Q. Zhang, H.-W. Lui, J. Am. Chem. Soc. 2000, 122, 9065–9070.
[13] R. R. Schmidt, B. Wegmann, K.-H. Jung, Liebigs Ann. Chem.
1991, 121–124.
[14] S. Zamyatina, C. Gronow, M. Oertelt, H. Puchberger, P. Brade,
P. Kosma, Angew. Chem. 2000, 112, 4322–4325; Angew. Chem.
Int. Ed. 2000, 39, 4150–4153.
[15] H. Gold, P. van Delft, N. Meeuwenoord, J. D. C. Code, D. V.
Filippov, G. Eggink, H. S. Overkleeft, G. A. van der Marel, J.
Org. Chem. 2008, 73, 9458–9460.
[16] M. Arlt, O. Hindsgaul, J. Org. Chem. 1995, 60, 14–15.
[17] S. C. Timmons, D. L. Jakeman, Org. Lett. 2007, 9, 1227–1230.
[18] P. Peltier, R. Daniellou, C. Nugier-Chauvin, V. Ferrières, Org.
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[19] R. Stiller, J. Thiem, Liebigs Ann. Chem. 1992, 467–471.
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[21] S. Warnecke, C. Meier, J. Org. Chem. 2009, 74, 3024–3030.
[22] C. Meier, Eur. J. Org. Chem. 2006, 1081–1102.
[23] S. Wendicke, S. Warnecke, C. Meier, Angew. Chem. 2008, 120,
1523–1525; Angew. Chem. Int. Ed. 2008, 47, 1500–1502.
[24] S. Wolf, T. Zismann, N. Lunau, C. Meier, Chem. Eur. J. 2009,
15, 7656–7664.
11.7 (CH3) ppm. 31P NMR (162 MHz, D2O): δ = –11.26 (d, JPP
=
19.8 Hz, Pα), –12.85 (d, JPP = 19.8 Hz, P ) ppm. IR (KBr): ν =
˜
β
3271, 2102, 1681, 1470, 1232, 1080, 1050, 915, 834, 508, 413 cm–1.
–
HRMS (ESI–): calcd. for C16H24N5O15P2 [M + H+] 588.0750;
found 588.0747. [α]2D4 = +8.7 (c = 0.1, H2O).
d4T-α-D-mannose 39: According to the general procedure, the reac-
tion was conducted with α-d-mannopyranosyl phosphate (99.3 mg,
0.28 mmol), 5-nitro-cycloSal-3Ј-deoxy-3Ј-dehydrothymidine mono-
phosphate, (60 mg, 0.14 mmol), and N,N-dimethylformamide
(4 mL) within 22 h to give 39 (52.4 mg, 0.10 mmol, 70%). 1H NMR
4
(400 MHz, D2O): δ = 7.47 (d, JHH = 4.1 Hz, 1 H, 6base-H), 6.99–
3
6.97 (m, 1 H, 3Ј-H), 6.52 (d, JHH = 6.1 Hz, 1 H, 2Ј-H), 5.96 (d,
3JHH = 6.1 Hz, 1 H, 1Ј-H), 5.50 (dd, JHP = 7.7, JHH = 1.3 Hz, 1
3
3
4
3
H, 1-H), 4.98 (dd, JHP = 8.0, JHH = 3.0 Hz, 1 H, 4Ј-H), 4.19–
4.11 (m, 2 H, 5Ј-H), 3.91–3.77 (m, 4 H, 2-H, 5-H, 6-H), 3.50–3.42
(m, 2 H, 3-H, 4-H), 1.92 (s, 3 H, CH3) ppm. 13C NMR (101 MHz,
D2O): δ = 166.3 (C4base), 151.9 (C2base), 138.2 (C6base), 134.3 (C3Ј),
125.3 (C2Ј),102.8 (C5base), 95.6 (d, 2JCP = 6.8 Hz, C1), 86.2 (d, 3JCP
[25] S. Wolf, T. Zismann, N. Lunau, S. Warnecke, S. Wendicke, C.
Meier, Eur. J. Cell Biol. 2010, 89, 63–75.
3
= 9.3 Hz, C2), 88.4 (C1Ј), 72.6 (C4), 71.7 (d, JCP = 7.6 Hz, C4Ј),
2
69.8 (C3), 69.4 (C5), 65.1 (d, JCP = 5.7 Hz, C5Ј), 60.5 (C6), 11.6
[26] M. Hoch, E. Heinz, R. R. Schmidt, Carbohydr. Res. 1989, 191,
(CH3) ppm. 31P NMR (162 MHz, D2O): δ = –11.66 (d, JPP
=
21–28.
19.8 Hz, Pα), –13.91 (d, JPP = 19.8 Hz, P ) ppm. IR (KBr): ν =
˜
β
[27] E. Heinz, H. Schmidt, M. Hoch, K. H. Jung, H. Binder, R. R.
Schmidt, Eur. J. Biochem. 1989, 184, 445–453.
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Chem. Soc. 1997, 119, 11743–11746.
3228, 2123, 1701, 1469, 1228, 917, 743, 512 cm–1. HRMS (ESI–):
calcd. for C16H23N2O15P2– [M + H+] 545.0579; found 545.0572. [α]
24
= –3.9 (c = 0.1, H2O).
D
Supporting Information (see footnote on the first page of this arti-
cle): 31P, 1H and 13C NMR spectra for compounds 4, 6, 24–27 and
35–39.
[30] G. Excoffier, D. Gagnaire, J. P. Utille, Carbohydr. Res. 1975, 39,
368–373.
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J. Org. Chem. 1966, 31, 205–211.
Acknowledgments
We thank Prof. Dr. E. Heinz for the generous gift of 6-deoxy-6-
sulfo-α-d-glucopyranosyl phosphate. Furthermore, we thank Prof.
Dr. J. Balzarini, Rega Institute for Medical Research, KU Leuven,
Belgium, for the antiviral tests of the NDP sugars with nucleoside
analogues.
[34] J. P. Horwitz, J. Chua, M. Noel, J. Org. Chem. 1964, 29, 2076–
2078.
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2817–2820.
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932–940.
Received: June 21, 2011
Published Online: September 8, 2011
[1] L. F. Leloir, Science 1971, 172, 1299–1303.
Eur. J. Org. Chem. 2011, 6304–6313
© 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
6313