The synthesis of new fluorinated or nonfluorinated sugar phosphonates and phosphoramidates as building blocks in the synthesis of modified hyaluronic acid subunits

Article abstract of DOI:10.1080/10426507.2017.1311332

The synthesis of several new fluorinated or nonfluorinated sugar phosphonates and phosphoramidates as building blocks for the synthesis of modified hyaluronic acid subunits is described. These compounds were prepared from d-glucose and d-glucosamine hydro

Full text of DOI:10.1080/10426507.2017.1311332

Phosphorus, Sulfur, and Silicon and the Related Elements
ISSN: 1042-6507 (Print) 1563-5325 (Online) Journal homepage: http://www.tandfonline.com/loi/gpss20
The synthesis of new fluorinated or
nonfluorinated sugar phosphonates and
phosphoramidates as building blocks in the
synthesis of modified hyaluronic acid subunits
Katarzyna Koroniak-Szejn, Joanna Tomaszewska & Henryk Koroniak
To cite this article: Katarzyna Koroniak-Szejn, Joanna Tomaszewska & Henryk Koroniak (2017):
The synthesis of new fluorinated or nonfluorinated sugar phosphonates and phosphoramidates
as building blocks in the synthesis of modified hyaluronic acid subunits, Phosphorus, Sulfur, and
Silicon and the Related Elements, DOI: 10.1080/10426507.2017.1311332
To link to this article: http://dx.doi.org/10.1080/10426507.2017.1311332
Accepted author version posted online: 27
Mar 2017.
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Date: 27 March 2017, At: 09:06

ACCEPTED MANUSCRIPT
The synthesis of new fluorinated or nonfluorinated sugar phosphonates and
phosphoramidates as building blocks in the synthesis of modified hyaluronic acid subunits
*
Katarzyna Koroniak-Szejn , Joanna Tomaszewska & Henryk Koroniak
Faculty of Chemistry, Adam Mickiewicz University, ul. Umultowska 89b, 60-614 Poznań,
Poland
*
Corresponding author E-mail address: katarzyna.koroniak@amu.edu.pl
In memoriam of Harry R. Hudson.
Abstract
The synthesis of several new fluorinated or nonfluorinated sugar phosphonates and
phosphoramidates as building blocks for the synthesis of modified hyaluronic acid subunits is
described. These compounds were prepared from D-glucose and D-glucosamine hydrochloride.
The syntheses of phosphonates and phosphoramidates are based on ARBUZOV and STAUDINGER
1
13
19
31
reactions. The products were fully characterized by H, C, F, P NMR and ESI MS
spectroscopy.
1
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Keywords
fluorinated sugar phosphonates, fluorinated sugar phosphoramidates, Arbuzov reaction,
neighbouring group participation
2
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1
. Introduction
Organophosphorus compounds, especially natural phosphates play an important role in
many biological processes. It is worth mentioning that glycosyl phosphates are of great interests
1
,2
by many researchers . These species are key glycosylating agents in biosynthesis of
oligosaccharides by glycosyl transferases. Also they can be used as metabolic intermediates or
3
–5
backbones for phosphorus-based biologically active compounds .
For many years, much attention has been focused on the synthesis of phosphonates and
6
,7
carbohydrate phosphonates as the analogues of phosphates . Furthermore, the phosphorus-
oxygen bond (P-O) is sensitive to hydrolysis by typical phosphate enzymes in contrast to the
6
,8
phosphorus-carbon bond (P-C) in phosphonates, which is hydrolytically stable . Moreover, the
replacement of an oxygen atom in phosphates by a methylene group changes the polarity and
9
acidity of the compound (Figure 1). Also in phosphonate analogues we can observe some
changes in bond length between C - C - P atoms (1.51 Å-1.80 Å -1.50 Å), which is slightly
6
,8
longer comparing to C - O - P bond (1.43 Å-1.59 Å-1.49 Å) .
Another example of phosphates analogues are phosphoramidates where instead of a
phosphorus-oxygen bond (P - O), a phosphorus-nitrogen bond (P - N) is present (Figure 2). This
class of organophosphorus compounds plays a dominant role in many processes.
1
0,11
12,13
Phosphoramidates have been used as catalysts
, inhibitors
and as a substitute for naturally
1
4,15
occurring phosphate group . Additionally, these compounds play an important role in
medicine and biological chemistry due to their biological activity. They have been used in the
3
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1
6,17
treatments of cancer and HIV
. Additionally, they display potent cytostatic, antiviral and
1
8
antifungal activities , which open up new inspirations for the synthesis of biologically active
analogues. Among them, glycosyl phosphoramidates are used as novel phosphoramidate
1
9
20
prodrugs or isosteric analogues of glycosyl phosphates .
Recently, special interest has been focused on the synthesis of drug delivery platforms.
One of the natural polymers, which can be successfully used in drug delivery applications is
hyaluronic acid (HA). This compound is a natural polysaccharide, which consists of the
repeating disaccharide subunits of β-D-N-acetylglucosamine and β-D-glucuronic acid (Figure 3).
HA is biodegradable and biocompatible, which makes this molecule a promising drug carrier.
There are several reports describing examples of modified HA-drug conjugate acting on cancer
2
1–23
cells
.
Diverse use of phosphonates and phosphoramidates shows, that they may be used
successfully as compounds displaying biological activity. The introduction of these two species
in building blocks of a drug delivery platform should be advantageous. It is also important, that
phosphonate and phosphoramidate moiety in combination with a fluorinated motif in the
molecule would lower polarity of the molecules. Because of that it is possible to increase the
lipophilicity of hyaluronic acid and increase the ability to penetrate cell membranes. Additionally
1
9
for such compounds, F NMR spectroscopy can be successfully used as an efficient tool for
2
4,25
studying the interactions between fluorinated compounds and biological systems
.
2
. Results and Discussion
4
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Two routes of conversion of fluorinated and nonfluorinated monosaccharides into sugar
phosphonates and sugar phosphoramidates have been explored. Our goal was to develop a
synthetic methodology for some model monosaccharides as building blocks for further HA
subunits synthesis. One of the common approaches for the formation of P - C bond is the
2
6
ARBUZOV reaction . In this reaction the nucleophilic triethyl phosphite P(OEt)
3
reacts in S 2
N
reaction with electrophylic alkyl halides (-Cl, -Br, -I). Alternatively, the STAUDINGER reaction
allows to obtain phosphoramidates with P - N bonds from azides and trivalent phosphorus e.g.
1
4,27
P(OEt)
3
.
In this paper we would like to present our results on the synthesis of new modified
monosaccharides based on D-glucose and D-glucosamine hydrochloride derivatives.
Our initial goal was the synthesis of the monosaccharides 2 and 6 with 2-hydroxyethyl
moieties, which has started from D-glucose 1 and D-glucosamine hydrochloride 5. This
2
8
procedure is known for 2-hydroxyethyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside 2 but only
2
9,30
partially for compound 6
. The sugar phosphonates 4 and 8 were obtained from corresponding
3
1,32
bromides 3
and 7 by heating at reflux with P(OEt) (Scheme 1). In several preparation of
3
3
2
phosphonates, like the one described by VALÉRY et al. triethyl phosphite was used in large
excess (even as a 156-fold excess). Such a huge excess of reagent can cause some problems with
isolation and purification of the product. In our case, evaporation of the reactant on rotavap
followed by column chromatography was not sufficient. The TLC analysis showed that our
3
1
products (4, 8) were more polar than P(OEt) itself. In P NMR spectra, except signals from the
3
main products 1-diethylphosphonoethyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside 4 or 1-
diethylphosphonoethyl-2-trifluoroacetamido-3,4,6-tri-O-acetyl-β-D-glucopyranoside 8, we have
5
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observed a lot of signals due to impurities and by-products. However, it was found that reduced
pressure distillation of excess of P(OEt) allows the complete removal of emerging by-products.
3
3
1
In P NMR spectrum only two signals were observed: 27.84 pm for phosphonate 4 and 28.53
ppm for phosphonate 8.
2
0
Sugar phosphoramidates 11 and 13 were prepared by Staudinger reaction from the
corresponding 2-azidoethyl 2,3,4,6-tetra-O-acetyl-β-D-glucose 10 and sugar azide having
trifluoroacetamide group 12 (Scheme 2) with high yields (93-94%). It is possible to use several
3
3
other methods for azide synthesis (i) in anhydrous DMF with heating in 60°C , in (ii) acetone
3
4
35
with KI at room temperature or (iii) using phase transfer catalysis (PTC) . The synthesis of 2-
azidoethyl 3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-β-D-glucopyranoside 12 was reported
3
0
in the literature previously. Generally azide 12 is prepared from bromide or sugar protected
3
6
with acetyl group in reaction with 2-azidoethanol in the presence of the Lewis acid.
Nucleophilic displacement reaction are often used in carbohydrate chemistry. In the
literature we can find reports on substitution reactions on anomeric carbon atoms in
monosaccharide with aliphatic alcohols. It is often observed that this kind of reaction proceeds
with neighboring group participation effect. In one of the stages of the reaction an oxonium ion
3
7
is formed, as a relatively stable five-member ring intermediate . In many cases formation of an
38,39
oxazoline ring was the main product
In our reaction with sugar 19 as a nucleophile, we have used partially fluorinated alcohols 14-18
Figure 4). The presence of the fluorine substituents will also lead to a dramatic change in
.
(
electronic properties of the molecule. In order to decrease the inductive effect caused by the
6
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fluorine substituents, it is necessary to use a short spacer between fluorinated carbon chain and
rest of the molecule (Figure 4). In our test reaction, when we have used a large excess of
1
2
,2,3,3,4,4,5,5-octafluoropentan-1-ol 15 like (16-, 8- or 4-fold excess), H NMR analysis of the
crude product showed the desired product and some by-products derived from the fluorinated
alcohol 15. Therefore, we have decided to limit the amount of fluorinated alcohol to only 1-2
equiv. of alcohol. It turned out that 1 equiv. can be used only for alcohol with two nonfluorinated
CH groups in 1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluorooctanol 17. In all other cases we have used 2
2
equiv. of fluorinated alcohols. In case of 14-16 and 18 the main product was 1-hydroxy-3,4,6-tri-
1
O-acetyl-2-chloroacetamido-2-deoxy-α-D-glucopyranose. In HNMR spectrum we have
3
observed a triplet at 5.31 ppm with coupling constant JH-H = 3.6 Hz. In reaction with 1 equiv. of
fluorinated alcohol 14-16 and 18, we have obtained only a traces of products, which was
confirmed only by a highly sensitive analysis ESI MS. For the reaction with 2,2,3,3,4,4,5,5-
+
octafluoropentan-1-ol 15 e.g. m/z [M+H] calcd for C19
H23ClF
8
NO : 596.0934; found: 596.0927;
9
+
m/z [M+Na] calcd for C19
H22ClF
8
NNaO
9
: 618.0753; found: 618.0745. We did not observed in
1
H NMR spectrum a signal from the CF
2
H group around 6.2 ppm. Unfortunately, for the reaction
with the short chain alcohol 1,1,1,3,3,3-hexafluoropropan-2-ol 14 we did not observe the desired
product neither in NMR nor MS spectra.
According to the literature, the synthesis of 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-
deoxy-β-D-glucopyranose 19 required introduction of the 2-chloroacetamide group, what was
4
0
41
42
described by FONDY et al. , WONG et al. or WITTE et al. . Compound 19 was prepared with
1
3
very good yield. Surprisingly, to the best of our knowledge we did not find C NMR and HRMS
spectra for this compound. The next set of compounds was prepared from 1,3,4,6-tetra-O-acetyl-
7
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2
-chloroacetamido-2-deoxy-β-D-glucopyranose 19 as a starting material. Sugar 19 reacts with
fluorinated alcohols 16-18 in the presence of BF ∙OEt as a Lewis acid. Reactant 19 itself can be
3
2
obtained from D-glucosamine hydrochloride 5 in a multi-steps sequence. The results of this
reaction are monosaccharides 20-22 with fluorinated aliphatic chains having 13, 15, 20 fluorine
atoms on substituent in anomeric position (Scheme 3). In the next step we wanted to eliminate
problems with excess of P(OEt)
compounds 23-25 we applied the reaction conditions described by LAKOUD et al. . These
authors reported the synthesis of phosphonates only with 5 equiv. of P(OEt) . However, we run
3
, that has been mentioned previously. In syntheses of
4
3
3
4
1
our experiment in mild conditions, which were described by WONG et al. . In some cases we
have used only 2 equiv. of P(OEt) , which was sufficient. The reaction was conducted with
3
heating in 65°C and in the presence of NaI or KI. These reaction conditions allowed us to
achieve high yields even for the fluorinated monosaccharides 23-25 with 68–86%. It turned out
that it is possible to use only 2 equiv. excess of triethyl phosphate in this kind of reaction that
facilitates further purification.
Next, we have turned our attention to the syntheses of phosphoramidates 29-31 starting from
the corresponding azides 26-28 (Scheme 3). The reactions were carried out according to the
2
0
procedure described before with excellent yields 76–92%.
Consequently we have studied the preparation of the nonfluorinated phosphonate 32, 34
and nonfluorinated phosphoramidate 36, 38 as the analogues of 2-chloroacetamido-1,2,4,6-tetra-
O-acetyl-2-deoxy-β-D-glucopyranose 19 and 2-bromopropionamido-1,2,4,6-tetra-O-acetyl-2-
deoxy-β-D-glucopyranose 33 (Scheme 4). These compounds were prepared as a model
nonfluorinated monosaccharides. The chloride 19 was converted under ARBUZOV conditions to 2-
8
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diethylphosphonoacetamid-1,2,4,6-tetra-O-acetyl-2-deoxy-β-D-glucopyranose 32 according to
4
1
the procedure described by WONG et al. with 69% yield. Similarly, from the bromide 33 we
have obtained 2-diethylphosphonopropionamid-1,2,4,6-tetra-O-acetyl-2-deoxy-β-D-
glucopyranose 34. The syntheses of 36 and 38 were accomplished in two-step reactions with
1% and 69% yields for 2-diethylphosphoramidoacetamid-1,2,4,6-tetra-O-acetyl-2-deoxy-β-D-
glucopyranose 36 and with 92 % and 60 % yields for 2-diethylphosphoramidopropionamid-
,2,4,6-tetra-O-acetyl-2-deoxy-β-D-glucopyranose 38 (Scheme 5). To the best of our knowledge
compounds 34, 36 and 38 are new compounds, first reported examples of this class.
9
1
3
. Conclusions
In summary, we have developed an efficient synthetic route to a library of modified
monosaccharides, which may be useful way in the synthesis of hyaluronic acid subunits. The
starting materials 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy-β-D-glucopyranose 19 and
2
-bromopropionamido-1,2,4,6-tetra-O-acetyl-2-deoxy-β-D-glucopyranose 33 were prepared with
good yields (87% and 75%) in easy multistage reaction. The synthesis of fluorinated
phosphonates and phosphoramidates containing in their structure aliphatic chains with 13, 15 and
2
1 fluorine atoms was optimized.
4
. Experimental
All chemicals were reagent grade and were used as purchased without further purification.
Anhydrous dichloromethane (DCM) was prepared by pre-drying with anhydrous CaCl
2
followed
9
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by distillation over CaH under argon atmosphere. All moisture sensitive reactions were carried
2
out under argon atmosphere. Thin-layer chromatography (TLC) was carried out on silica gel
plates (Silica gel 60, F254, Merck) with detection by UV light or with stain solution (Cerium
Molybdate Stain and Potassium Permanganate). Purification was performed with column
1
13
19
chromatography using normal-phase silica gel (Silica gel 60, 70-230 mesh, Fluka). H, C, F,
3
1
P NMR were recorded with a Varian VNMR-S 400 MHz, VARIAN Mercury 300 MHz, Varian
GEMINI 300 (300 MHz) or Bruker Avance 400 (400 MHz), Bruker ASCEND 600 (600 MHz)
spectrometers. Chemical shifts are reported as δ values (ppm). NMR spectra were recorded in
1
13
deuterated solvents and calibrated using an internal reference: TMS ( H), CDCl
3
( C), CFCl
3
1
9
31
(
F) and 85% H
3
PO
4
( P) as an external standard (δ = 0 ppm). Multiplicities of signals are
described as follows: s = singlet, d = doublet, dd = doublet of doublets, ddd = doublet of doublet
of doublets, t = triplet, td = triplet of doublets, tt = triplet of triplets, m = multiplet. Coupling
constants (J) are given in Hz. Mass spectra (ESI) were carried out with Q-TOF (Impact HD,
Bruker) and Agilent 1200 LC system with Agilent Q-TOF 6540 spectrometer with DUAL AJS
1
13
19
31
ESI source. The Supplemental Materials contains some selected H, C, F and P NMR
spectra of products. (Figures S 1 – S 23)
4
.1. 1-Diethylphosphonoethyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside 4
A mixture of bromide 3 (0.104 g, 0.229 mmol) and P(OEt) (5.90 mL, 35.720 mmol) was heated
under reflux for 15 h. The excess of P(OEt) was distilled off under reduced pressure and the
3
3
crude product was purified by column chromatography (DCM/MeOH 20:1, v/v) afforded 4 as an
1
oil. Yield: (0.097 g) 83%; H NMR (400 MHz, CDCl
3
): δ 5.19 (appt, J = 9.5 Hz, 1H, H-3), 5.06
(
t, J = 9.7 Hz, 1H, H-4), 4.96 (dd, J = 8.0 Hz, J = 9.5 Hz, 1H, H-2), 4.52 (d, J = 8.0 Hz, 1H, H-
10
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1
), 4.26 (dd, J = 4.8 Hz, J = 12.3 Hz, 1H, H-6a), 4.14-4.00 (m, 6H, H-6b, 2xOCH
OCH CH ), 3.85-3.74 (m, 1H, OCH CH ), 3.69 (ddd, J = 2.4 Hz, J = 4.8 Hz, J = 9.9 Hz, 1H, H-
), 2.17-2.09 (m, 2H, CH P(O)), 2.08 (s, 3H, OAc), 2.04 (s, 3H, OAc), 2.01 (s, 3H, OAc), 1.99
s, 3H, OAc), 1.35-1.29 (m, 6H, 2xCH
C(O)CH , 169.4 C(O)CH , 169.3 C(O)CH
4.2 C-6, 63.6 (d, J = 5.8 Hz, OCH CH P(O)(OEt)
J = 6.3 Hz, OCH CH ), 27.0 (d, J = 139.3 Hz, OCH
C(O)CH , 20.57 2xC(O)CH , 16.4 (d, J = 6.0 Hz, OCH
P NMR (121 MHz, CDCl
2
CH
3
,
b
2
a
2
5
2
1
3
(
3
); C NMR (101 MHz, CDCl
, 100.7 C-1, 72.8 C-5, 71.9 C-2, 71.2 C-3, 68.3 C-4,
), 61.84 (d, J = 5.4 Hz, OCH CH ), 61.72 (d,
CH P(O)(OEt) ), 20.69 C(O)CH , 20.66
CH ), 16.1 (d, J = 6.7 Hz, OCH CH );
3
): δ 170.6 C(O)CH
3
, 170.2
3
3
3
6
2
2
2
2
3
2
3
2
2
2
3
3
3
2
3
2
3
3
1
+
3
): δ 27.84 (s, 1P); MS (ESI) m/z [M+H] calcd for C20
H O13P:
34
+
5
4
13.1737; found: 513.1737 m/z [M+Na] calcd for C20
H
33NaO13P: 535.1556; found: 535.1557.
.2. 2-Hydroxyethyl 3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-β-D-glucopyranoside 6
To a solution of 3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-α-D-glucopyranosyl bromide
0.514g, 1.110 mmol) in anhydrous DCM (28.40 mL) was added anhydrous Na SO and 1,2-
ethanediol (0.62 mL, 11.100 mmol). After stirring at room temperature for 15 min, under an
argon atmosphere, Ag CO (4.60 g, 16.700 mmol) was added to the reaction mixture and stirred
overnight protected from the light. The reaction mixture was filtered, washed with water (3x30
mL), dried over Na SO and the solvent was evaporated. The crude product was purified by
(
2
4
2
3
2
4
column chromatography using a 5:1 mixture of EtOAc/cyclohexane (CH) as eluent. Compound 6
1
was obtained as a white solid. Yield: (0.299 g) 60%; H NMR (400 MHz, CDCl
3
): δ 7.39 (d, J =
8
.8 Hz, 1H, NH), 5.30 (dd, J = 9.4 Hz, J = 10.6 Hz, 1H, H-3), 5.10 (t, J = 9.7 Hz, 1H, H-4), 4.75
d, J = 9.7 Hz, 1H, H-1), 4.26 (dd, J = 5.2 Hz, J = 12.3 Hz, 1H, H-6a), 4.20 (dd, J = 2.6 Hz, J =
2.3 Hz, 1H, H-6b), 4.07 (dd, J = 8.7 Hz, J = 19.2 Hz, 1H, H-2), 3.90 (ddd, J = 2.4 Hz, J = 4.7
(
1
11
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Hz, J = 9.9 Hz, 1H, H-5), 3.84-3.72 (m, 4H, OCH
2
CH OH), 2.83 (t, J = 5.2 Hz, 1H, OH), 2.11
2
1
3
(
s, 3H, OAc), 2.06 (s, 3H, OAc), 2.05 (s, 3H, OAc); C NMR (101 MHz, CDCl
C(O)CH , 170.8 C(O)CH , 169.4 C(O)CH , 157.7 (q, J = 37.8 Hz, C(O)CF ), 115.6 (q, J = 289.1
Hz, CF ), 101.0 C-1, 72.4 C-5, 72.0 C-3, 71.9 C-4, 68.4 OCH CH OH, 62.0 C-6, 61.7
OCH CH
CDCl
3
): δ 171.1
3
3
3
3
3
2
2
1
9
2
2
OH, 54.8 C-2, 20.7 C(O)CH
3
, 20.6 C(O)CH
3
, 20.4 C(O)CH
3
; F NMR (376 MHz,
+
3
): δ -76.03 (s, 3F, CF
3
); MS (ESI) m/z [M+H] calcd for C16
H
F
23 3
NO10: 446.1275; found:
+
4
4
46.1271 m/z [M+Na] calcd for C16
H
22
F NNaO10: 468.1094; found: 468.1101.
3
.3. 2-Bromoethyl 3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-β-D-glucopyranoside 7
To a solution of alcohol 6 (0.124 g, 0.279 mmol) in anhydrous DCM (3.50 mL) were added PPh
0.146 g, 0.558 mmol) and CBr (0.204 g, 0.614 mmol) under an argon atmosphere. After
stirring for 2 h at room temperature, the reaction mixture was diluted with DCM (11 mL) and
washed with water (2x12 mL). The organic layer was then dried over Na SO and concentrated.
3
(
4
2
4
Purification by column chromatography (CH/EtOAc 2:1,v/v) afforded 7 as a white solid. Yield:
1
(
0.105 g) 75%; H NMR (300 MHz, CDCl
3
): δ 6.66 (d, J = 8.6 Hz, 1H, NH), 5.31 (dd, J = 9.3
Hz, J = 10.7 Hz, 1H, H-3), 5.11 (t, J = 9.6 Hz, 1H, H-4), 4.77 (d, J = 8.3 Hz, 1H, H-1), 4.28 (dd,
J = 4.8 Hz, J = 12.4 Hz, 1H, H-6a), 4.24-4.13 (m, 2H, H-6b, OCH CH Br), 4.02 (dd, J = 8.7 Hz,
J = 19.3 Hz, 1H, H-2), 3.87-3.72 (m, 2H, H-5, OCH CH Br), 3.40-3.44 (m, 2H, CH Br), 2.11 (s,
H, OAc), 2.05 (s, 3H, OAc), 2.04 (s, 3H, OAc); C NMR (75 MHz, CDCl ): δ 171.0 C(O)CH
70.6 C(O)CH , 169.3 C(O)CH , 157.4 (q, J = 37.5 Hz, C(O)CF ), 115.5 (q, J = 286.3 Hz, CF ),
00.4 C-1, 72.1 C-5, 71.5 C-3, 69.7 C-4, 68.2 OCH CH Br, 61.8 C-6, 54.9 C-2, 30.0
CH Br, 20.7 C(O)CH , 20.6 C(O)CH , 20.4 C(O)CH
a
2
b
2
2
1
3
3
1
1
3
3
,
3
3
3
3
2
2
1
9
OCH
2
2
3
3
3
; F NMR (282 MHz, CDCl ): δ -
3
12
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+
7
6.50 (s, 3F, CF
3
); MS (ESI) m/z [M+H] calcd for C16
H22BrF
3
NO
9
: 508.0431; found: 508.0501
+
m/z [M+Na] calcd for C16
H21BrF
3
NNaO
9
: 530.0249; found: 530.0250.
4
.4. 1-Diethylphosphonoethyl-2-trifluoroacetamido-3,4,6-tri-O-acetyl-β-D-glucopyranoside 8
A mixture of bromide 7 (0.099 g, 0.194 mmol) and P(OEt) (5.20 mL, 30.260 mmol) was heated
under reflux for 15 h. The excess of P(OEt) was distilled off under reduced pressure and crude
3
3
product was purified by column chromatography (DCM/MeOH 20:1, v/v) afforded 8. Yield:
1
(
0.094 g) 85%; H NMR (400 MHz, CDCl
3
): δ 8.56 (d, J = 8.4 Hz, 1H, NH), 5.23 (appt, J = 9.9
Hz, 1H, H-3), 5.11 (t, J = 9.6 Hz, 1H, H-4), 4.82 (d, J = 8.5 Hz, 1H, H-1), 4.29 (dd, J = 4.5 Hz, J
12.3 Hz, 1H, H-6a), 4.18-4.03 (m, 7H, H-6b, OCH CH , 2xOCH CH , H-2), 3.91-3.78 (m, 1H,
OCH CH ), 3.71 (ddd, J = 2.4 Hz, J = 4.3 Hz, J = 9.9 Hz, 1H, H-5), 2.10 (s, 3H, OAc), 2.10-2.02
m, 2H, CH
101 MHz, CDCl
), 115.9 (q, J = 288.0 Hz, CF
Hz, OCH CH P(O)(OEt) ), 62.3 (d, J = 6.5 Hz, OCH
OCH CH ), 54.1 C-2, 26.4 (d, J = 141 Hz, OCH CH
C(O)CH , 20.4 C(O)CH , 16.38 (d, J = 6.1 Hz, OCH CH ), 16.25 (d, J = 6.0 Hz, OCH
=
b
2
2
3
a
2
1
3
(
(
2
P(O)), 2.02 (s, 3H, OAc), 2.01 (s, 3H, OAc), 1.37-1.31 (m, 6H, 2xCH
): δ 170.7 C(O)CH , 170.6 C(O)CH , 169.3 C(O)CH , 157.8 (q, J = 37.4 Hz,
), 99.9 C-1, 72.9 C-5, 72.1 C-3, 68.4 C-4, 63.7 (d, J = 5.7
CH ), 61.9 C-6, 61.8 (d, J = 6.8 Hz,
P(O)(OEt)
3
); C NMR
3
3
3
3
C(O)CF
3
3
2
2
2
2
3
2
3
2
2
2
), 20.7 C(O)CH
3
, 20.6
1
9
3
3
2
3
2
CH ); F
3
3
1
NMR (282 MHz, CDCl
3
): δ -76.21 (s, 3F, CF
3
); P NMR (121 MHz, CDCl
3
): δ 28.53 (s, 1P);
+
+
MS (ESI) m/z [M+H] calcd for C20
H
32
F
3
NO12P: 566.1614; found: 566.1615 m/z [M+Na] calcd
for C20
H
31
F
3
NNaO12P: 588.1434; found: 588.1434.
4
.5. 1-Diethylphosphoramidoethoxy-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside 11
The mixture of azide 10 (0.080 g, 0.192 mmol) in anhydrous DCM (8.20 mL) with powdered
molecular sieves 4Å (0.017 g) was stirred for 1h under an argon atomosphere at room
13
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temperature. Then P(OEt) was added (0.16 mL, 0.960 mmol) and stirred overnight. The reaction
3
mixture was filtered through a pad of Celite and concentrated under reduced pressure.
Purification by column chromatography (DCM/MeOH 20:1,v/v) afforded 11 as a white foam.
1
Yield: (0.095 g) 94%; H NMR (400 MHz, CDCl
3
): δ 5.22 (t, J = 9.3 Hz, 1H, H-3), 5.08 (appt, J
=
9.7 Hz, 1H, H-4), 4.99 (dd, J = 8.0 Hz, J = 9.6 Hz, 1H, H-2), 4.53 (d, J = 8.0 Hz, 1H, H-1),
.26 (dd, J = 4.8 Hz, J = 12.4 Hz, 1H, H-6a), 4.14 (dd, J = 2.4 Hz, J = 12.4 Hz, 1H, H-6b), 4.09-
.02 (m, 5H, 2xOCH CH , NHP(O)), 3.92-3.83 (m, 1H, OCH CH ), 3.71 (ddd, J = 2.4 Hz, J =
.7 Hz, J = 9.9 Hz, 1H, H-5), 3.64 (m, 1H, OCH CH ), 3.18-3.06 (m, 2H, OCH CH ), 2.10 (s,
H, OAc), 2.07 (s, 3H, OAc), 2.03 (s, 3H, OAc), 2.01 (s, 3H, OAc), 1.35-1.29 (m, 6H, 2xCH );
, 169.35
, 101.0 C-1, 72.7 C-5, 71.9 C-2, 71.3 C-3, 70.7 (d, J = 5.4 Hz,
4
4
4
3
2
3
a
2
b
2
2
2
3
1
3
C NMR (101 MHz, CDCl
3
): δ 170.6 C(O)CH
3
, 170.2 C(O)CH
3
, 169.41 C(O)CH
3
C(O)CH
3
OCH
2
2
CH
2
2
NHP(O)(OEt)
2
2
), 68.3 C-4, 62.3 (d, J = 5.5 Hz, OCH
2
CH ), 61.8 C-6, 41.1
3
OCH
CH
NHP(O)(OEt)
, 20.7 C(O)CH , 20.63 C(O)CH , 20.55 2xC(O)CH
3
3
3
, 16.2 (d, J = 7.0
3
1
+
Hz, 2xOCH
2
CH
3
); P NMR (121 MHz, CDCl
3
): δ 9.50 (s, 1P); MS (ESI) m/z [M+H] calcd for
+
C
20
H
36NO13P: 529.1925; found: 529.1873 m/z [M+Na] calcd for C20
H
35NNaO13P: 551.1744;
found: 551.1691.
.6. 2-Azidoethyl 3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-β-D-glucopyranoside 12
To solution of 2-bromoethyl 3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-β-D-
glucopyranoside 7 (0.082 g, 0.162 mmol) in DCM (0.82 mL) and aq. sat. NaHCO (0.82 mL),
TBASH (0.051 g, 0.149 mmol) and NaN (0.031 g, 0.483 mmol) were added. The reaction
mixture was stirred at room temperature for 48 h. It was subsequently diluted with EtOAc (5
mL) and washed with H O (8 mL) and aq. sat. NaHCO (8 mL), dried over Na SO and
4
a
3
3
2
3
2
4
14
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concentrated in vacuo. The crude product was purified by column chromatography (CH/EtOAc
1
9
1
:1, v/v) and afforded 12 as a white solid. Yield: (0.066 g) 87%; F NMR (282 MHz, CDCl
76.63 (s, 3F, CF ).
.7. 1-Diethylphosphoramidoethoxy-3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-β-D-
3
): δ
-
3
4
glucopyranoside 13
The mixture of azide 12 (0.055 g, 0.117 mmol) in anhydrous DCM (5.10 mL) with powdered
molecular sieves 4Å (0.025 g) was stirred for 1 h under an argon atmosphere at room
temperature. Then P(OEt) was added (0.1 mL, 0.585 mmol) and stirred overnight. The reaction
3
mixture was filtered through a pad of Celite and concentrated under reduced pressure.
Purification by column chromatography (DCM/MeOH 20:1,v/v) afforded 13 as a white solid.
1
Yield: (0.063 g) 93%; H NMR (400 MHz, CDCl
3
): δ 8.55 (d, J = 8.6 Hz, 1H, NH), 5.32 (dd, J =
9
.4 Hz, J = 10.5 Hz, 1H, H3), 5.10 (t, J = 9.7 Hz, 1H, H-4), 4.85 (d, J = 8.4 Hz, 1H, H-1), 4.28
dd, J = 4.6 Hz, J = 12.3 Hz, 1H, H-6a), 4.19-3.97 (m, 6H, H-6b, H-2, 2xOCH CH ), 3.95-3.86
m, 1H, OCH CH ), 3.73 (ddd, J = 2.4 Hz, J = 4.5 Hz, J = 10.0 Hz, 1H, H-5), 3.67-3.53 (m, 2H,
CH2, NHP(O)), 3.13-3.04 (m, 2H, OCH CH ), 2.11 (s, 3H, OAc), 2.03 (s, 3H, OAc), 2.02
); C NMR (101 MHz, CDCl ): δ 170.7 C(O)CH , 170.6
, 157.8 (q, J = 37.5 Hz, C(O)CF ), 115.8 (q, J = 289.3 Hz, CF ), 100.0
C-1, 72.1 C-5, 72.0 C-3, 70.0 (d, J = 5.3 Hz, OCH CH NHP(O)(OEt) ), 68.5 C-4, 62.61 (d, J =
.5 Hz, OCH CH ), 62.54 (d, J = 4.4 Hz, OCH CH ), 62.0 C-6, 54.7 C-2, 41.0
NHP(O)(OEt) , 20.4 C(O)CH , 16.2 (d, J = 6.9 Hz,
), 16.0 (d, J = 7.4 Hz, OCH ); F NMR (282 MHz, CDCl ): δ -76.22 (s, 3F, CF );
(
(
2
3
b
2
OCH
s, 3H, OAc), 1.37-1.29 (m, 6H, 2xCH
C(O)CH , 169.4 C(O)CH
a
2
2
1
3
(
3
3
3
3
3
3
3
2
2
2
4
2
3
2
3
OCH
2
2
CH
2
3
2
, 20.7 C(O)CH
3
, 20.6 C(O)CH
3
3
1
9
OCH
CH
2
CH
3
3
3
15
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3
1
+
P NMR (121 MHz, CDCl
3
): δ 9.33 (s, 1P); MS (ESI) m/z [M+H] calcd for C20
H
33
F
3
2
N O12P:
+
5
4
81.1724; found: 581.1737 m/z [M+Na] calcd for C20
H
32
F
3
2
N O12P: 603.1543; found: 603.1555.
.8. 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy- β-D-glucopyranoside 19
N (0.61 mL) was added to a solution of 2-deoxy-2-amino-1,3,4,6-tetra-O-acetyl-β-D-
glucopyranosyl hydrochloride (0.844 g, 2.200 mmol) in anhydrous DCM (10 mL) at 0 °C. After
min chloroacetic anhydride (0.640 g, 3.740 mmol) was added. The reaction mixture was stirred
Et
3
5
for 3.5 hours at room temperature and controlled by TLC. Next, the solution was diluted with
DCM (50 mL), washed successively with 1N HCl (2 x 50 mL) and brine (50 mL). The organic
phase was dried over Na
crystallized from EtOAc/CH giving 19 as a light grey and white solid. Yield: (0.807 g) 87%; C
NMR (101 MHz, CDCl ): δ 170.9 C(O)CH , 170.7 C(O)CH , 169.3 2xC(O)CH , 166.6
Cl, 20.9
2
SO
4
and solvent was evaporated. The resulting crude product was
1
3
3
3
3
3
C(O)CH
2
3
Cl, 92.1 C-1, 72.9 C-5, 71.9 C-3, 67.9 C-4, 61.7 C-6, 53.3 C-2, 42.3 CH
2
+
C(O)CH
, 20.7 C(O)CH
3
, 20.58 C(O)CH
3
, 20.57 C(O)CH
3
; MS (ESI) m/z [M+H] calcd for
+
C
16
H
23ClNO10: 424.1010; found: 424.1011 m/z [M+Na] calcd for C16
H
22ClNNaO10: 446.0830;
found: 446.0843.
.9. 3,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6,7,7-
4
pentadecafluoromethoxy)-β-D-glucopyranoside 20
To a stirred solution of 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy-β-D-glucopyranose 19
(
0.048 g, 0.113 mmol) in anhydrous DCM (0.23 mL) under an argon atmosphere and room
temperature BF ∙Et (0.03 mL, 0.226 mmol) and 1,1,1,2,2,3,3,4,4,5,5,6,6,7,7-
pentadecafluorooctanol 16 (0.090 g, 0.226 mmol) were added and stirred for 24 h. The reaction
mixture was diluted with EtOAc (2 mL) and washed with aq. sat. NaHCO (2mL). The organic
3
2
O
3
16
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phase was dried over Na
2
SO and concentrated in vacuo. The crude product was purified by
4
column chromatography (EtOAc/CH 1:1, v/v) afforded 20 as a white solid. Yield: (0.052 g)
1
6
0%; H NMR (400 MHz, CDCl
3
): δ 6.75 (d, J = 8.5 Hz, 1H, NH), 5.44 (appt, J = 9.9 Hz, 1H,
H-3), 5.10 (t, J = 9.5 Hz, 1H, H-4), 4.94 (d, J = 8.2 Hz, 1H, H-1), 4.37-4.25 (m, 2H, H-6a,H-6b),
4
2
.21-4.02 (m, 2H, OCH
.5 Hz, J = 4.6 Hz, J = 9.8 Hz, 1H, H-5), 2.10 (s, 3H, OAc), 2.04 (s, 6H, 2xOAc); C NMR
) δ 170.6 C(O)CH , 170.5 C(O)CH , 169.3 C(O)CH , 166.5 C(O)CH Cl,
22.7-106.8 (m, (CF CF ), 100.4 C-1, 72.2 C-5, 71.0 C-3, 68.2 C-4, 65.3 (t, J = 26.2 Hz,
(CF CF ), 61.7 C-6, 54.8 C-2, 42.2 CH Cl, 20.64 C(O)CH , 20.56 C(O)CH , 20.53
2
(CF
2
)
6
CF
3
), 3.98 (s, 2H, CH Cl), 3.93-3.84 (m, 1H, H-2), 3.78 (ddd, J =
2
1
3
(
101 MHz, CDCl
3
3
3
3
2
1
2
)
6
3
OCH
C(O)CH
2
2
)
6
3
2
3
3
1
9
3
; F NMR (376 MHz, CDCl
3
) δ: -80.77, -119.86, -122.02, -122.73, -123.13, -126.10;
+
+
MS (ESI) m/z [M+H] calcd for C22
H
22ClF15NO
9
: 764.0744; found: 764.0734; m/z [M+Na]
: 786.0563; found: 786.0559.
.10. 3,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6-
calcd for C22
H
21ClF15NNaO
9
4
tridecafluoroethoxy)-β-D-glucopyranoside 21
To a stirred solution of 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy-β-D-glucopyranose 19
(
0.091 g, 0.214 mmol) in anhydrous DCM (4.3 mL) under an argon atmosphere and room
temperature BF ∙Et O (0.06 mL, 0.428 mmol) and 1,1,1,2,2,3,3,4,4,5,5-pentadecafluorooctanol
7 (0.076 g, 0.214 mmol) were added and stirred for 24 h. The reaction mixture was diluted with
EtOAc (3 mL) and washed with aq. sat. NaHCO (3 mL). The organic phase was dried over
Na SO and concentrated in vacuo. The crude product was purified by column chromatography
EtOAc/CH 1:1, v/v) gave 21 as a white solid. Yield: (0.106 g) 69%; H NMR (300 MHz,
CDCl ) δ 6.65 (d, J = 8.6 Hz, 1H, NH), 5.40 (dd, J = 10.5 Hz, J = 9.3 Hz, 1H, H-3), 5.09 (t, J =
3
2
1
3
2
4
1
(
3
17
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9
.6 Hz, 1H, H-4), 4.83 (d, J = 8.2 Hz, 1H, H-1), 4.29 (dd, J = 12.3 Hz, J = 4.9 Hz, 1H, H-6a),
.21-4.12 (m, 2H, H-6b, H-2), 4.00 (d, J = 2.7 Hz, 2H, CH Cl), 3.91-3.82 (m, 2H,
CH (CF CF ), 3.77 (ddd, J = 9.9 Hz, J = 4.9 Hz, J = 2.4 Hz, 1H, H-5), 2.53-2.37 (m, 2H,
(CF CF ), 2.11 (s, 3H, OAc), 2.05 (s, 3H, OAc), 2.06 (s, 3H, OAc); C NMR (101 MHz,
CDCl ) δ 170.60 C(O)CH , 170.58 C(O)CH , 169.4 C(O)CH , 166.4 C(O)CH Cl, 121.7-106.4
m, (CF CF ), 100.4 C-1, 72.0 C-5, 71.4 C-3, 68.4 C-4, 61.9 C-6, OCH CH (CF CF , 55.0 C-
, 42.3 CH Cl, 31.3 (t, J = 21.5 Hz, OCH CH (CF CF ), 20.63 C(O)CH , 20.60 C(O)CH
0.55 C(O)CH ; F NMR (377 MHz, CDCl ) δ -80.77, -113.39, -121.89, -122.86, -123.53, -
: 728.0932; found: 728.0930; m/z
4
2
OCH
2
2
2
)
5
3
1
3
CH
2
2
)
5
3
3
3
3
3
2
(
2
)
5
3
2
2
2
)
5
3
2
2
1
2
2
2
2
)
5
3
3
3
,
1
9
3
3
+
26.12; MS (ESI) m/z [M+H] calcd for C22
H
24ClF13NO
9
+
[
M+Na] calcd for C22
H
23ClF13NNaO : 750.0751; found: 750.0748.
9
4
.11. 3,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-
(
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10-eicosafluorononylmethoxy)-β-D-glucopyranoside 22
To a stirred solution of 1,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy-β-D-glucopyranose 19
0.059 g, 0.139 mmol) in anhydrous DCM (0.60 mL) under an argon atmosphere and room
temperature, BF ∙Et O (0.17 mL, 1.390 mmol) and 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9-
(
3
2
eicosafluoroundecanol 18 (0.074 g, 0.139 mmol) gave 22 as a white solid. Yield: (0.081 g) 55%;
1
H NMR (403MHz, CDCl
H), 5.43 (dd, J = 10.6 Hz, J = 9.2 Hz, 1H, H-3), 5.10 (t, J = 9.5 Hz, 1H, H-4), 4.93 (d, J = 8.2
Hz, 1H, H-1), 4.34-4.23 (m, 3H, H-6a, OCH (CF CF H), 4.18 (dd, J = 12.4 Hz, J = 2.5 Hz, 1H,
H-6b), 3.99 (apps, 2H, CH Cl), 3.87 (m, 1H, H-2), 3.77 (ddd, J = 9.9 Hz, J = 4.9 Hz, J = 2.4 Hz,
H, H-5), 2.10 (s, 3H, OAc), 2.04 (s, 6H, 2xOAc); C NMR (101 MHz, CDCl
C(O)CH , 170.5 C(O)CH , 169.3 C(O)CH , 166.5 C(O)CH Cl, 118.2-105.8 (m, (CF
3
) δ 6.70 (d, J = 8.5 Hz, 1H, NH), 6.06 (tt, J = 5.0 Hz, J = 51.9 Hz, 1H,
CF
2
2
2
)
9
2
2
1
3
1
3
) δ 170.6
CF H),
3
3
3
2
2
)
9
2
18
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1
2
1
8
9
4
00.3 C-1, 72.2 C-5, 70.9 C-3, 68.2 C-4, 65.3 (t, J = 25.9, OCH
2
(CF
2
)
9
CF
2
H), 61.7 C-6, 54.9 C-
) δ -
1
9
, 42.2 CH
2
Cl, 20.7 C(O)CH
3
, 20.64 C(O)CH
3
, 20.58 C(O)CH
3
; F NMR (379 MHz, CDCl
3
+
20.33, -122.35, -123.70, -129.74, -137.47; MS (ESI) m/z [M+H] calcd for C25
H
23ClF20NO :
9
+
96.0742; found: 896.0731; m/z [M+Na] calcd for C25
H22ClF20NNaO : 918.0561; found:
9
18.0558.
.12. General procedure for the synthesis of fluorinated phosphonates 23, 24, 25
To a solution of sugar with fluorinated aliphatic chain 20, 21, 22 (1 equiv.) in MeCN (29.70 mL
for 1 mmol of sugar), KI (2.26 equiv.) and P(OEt) (5 equiv.) were added. The reaction mixture
3
was heated in 65 °C for 4 h. Solvent was evaporated and the crude product was purified by
column chromatography using a 20:1 mixture of DCM/MeOH as eluent to afforded compounds
2
3, 24, 25.
4
.12.1. 3,4,6-Tri-O-acetyl-2-deoxy-2-diethylphosphonoacetamido-1-
(
1,1,1,2,2,3,3,4,4,5,5,6,6,7,7-pentadecafluoromethoxy)-β-D-glucopyranoside 23
,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6,7,7-
3
pentadecafluoromethoxy)-β-D-glucopyranoside 20 (0.069 g, 0.090 mmol) and P(OEt)
3
(0.08 mL,
) δ 7.05
1
0
.450 mmol) gave 23 as a white solid. Yield: (0.062 g) 80%; H NMR (300 MHz, CDCl
3
(
d, J = 8.2 Hz, 1H, NH), 5.44 (dd, J = 10.5 Hz, J = 9.1 Hz, 1H, H-3), 5.10 (t, J = 9.5 Hz, 1H, H-
), 5.00 (d, J = 8.2 Hz, 1H, H-1), 4.28 (dd, J = 4.8 Hz, J = 12.3 Hz, 1H, H-6a), 4.23-3.95 (m, 7H,
OCH (CF CF , H-6b, 2xOCH CH ), 3.83-3.72 (m, 2H, H-2, H-5), 2.78 (d, J = 21.6 Hz, 2H,
CH P(O)), 2.09 (s, 3H, OAc), 2.05 (s, 3H, OAc), 2.03 (s, 3H, OAc), 1.38-1.29 (m, 6H, 2xCH );
C NMR (101 MHz, CDCl ) δ 170.6 C(O)CH , 170.5 C(O)CH , 169.5 C(O)CH , 169.4 (d, J =
.4 Hz, C(O)CH P(O)(OEt) ), 122.0-106.4 (m, (CF CF ), 100.5 C-1, 72.0 C-5, 71.2 C-3, 68.8
4
2
2
)
6
3
2
3
2
3
1
3
3
3
3
3
3
2
2
2
)
6
3
19
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C-4, 62.4 C-6, 61.9 (d, J = 6.6 Hz, OCH
OCH (CF CF ), 54.6 C-2, 34.4 (d, J = 129.2 Hz, C(O)CH
C(O)CH , 20.3 C(O)CH , 16.2 (d, J = 4.8 Hz, OCH CH ), 16.1 (d, J = 4.4 Hz, OCH
NMR (282 MHz, CDCl
2
CH
3
), 61.6 (d, J = 6.4 Hz, OCH
P(O)(OEt) ), 20.6 C(O)CH
CH ); F
2
CH
3
), 64.6 (appm,
2
2
)
6
3
2
2
3
, 20.4
1
9
3
3
2
3
2
3
3
1
3
) δ: -81.23, -120.56, -122.51, -123.23, -123.70, -126.62; P NMR (121
+
MHz, CDCl
3
): δ 22.53 (s, 1P); MS (ESI) m/z [M+H] calcd for C26
32
H F15NO12P: 866.1423;
+
found: 886.1430 m/z [M+Na] calcd for C26
31
H F15NNaO12P: 888.1242; found: 888.1253.
4
.12.2. 3,4,6-Tri-O-acetyl-2-deoxy-2-diethylphosphonoacetamido-1-(1,1,1,2,2,3,3,4,4,5,5,6,6-
tridecafluoroethoxy)-β-D-glucopyranoside 24
,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluoroethoxy)-
3
β-D-glucopyranoside 21 (0.105 g, 0.144 mmol) and P(OEt)
3
(0.05 mL, 0.288 mmol) gave 24 as a
) δ 6.93 (d, J = 8.2 Hz, 1H, NH),
1
white solid. Yield: (0.103 g) 86 %; H NMR (403MHz, CDCl
3
5
.38 (dd, J = 9.2 Hz, J = 10.5 Hz, 1H, H-3), 5.03 (t, J = 9.6 Hz, 1H, H-4), 4.84 (d, J = 8.3 Hz,
H, H-1), 4.26 (dd, J = 5.0 Hz, J = 12.3 Hz, 1H, H-6a), 4.17-4.06 (m, 6H, H-6b, 2xOCH CH
CH (CF CF ), 3.75 (ddd, J = 9.9 Hz, J = 4.9 Hz, J = 2.4 Hz, 1H,
H-5), 2.79 (d, J = 21.0 Hz, 2H, CH P(O)), 2.53-2.38 (m, 2H, CH (CF CF ), 2.08 (s, 3H, OAc),
.04 (s, 3H, OAc), 2.02 (s, 3H, OAc), 1.33 (td, J = 1.7 Hz, J = 7.1 Hz, 6H, 2xCH
101 MHz, CDCl ) δ 170.6 C(O)CH , 170.4 C(O)CH , 169.5 C(O)CH , 164.4 (d, J = 3.3 Hz,
P(O)(OEt) ), 122.0-106.7 (m, (CF CF ), 100.5 C-1, 71.8 C-5, 71.7 C-3, 68.8 C-4,
CH ), 62.7 (d, J = 7.1 Hz, OCH CH ), 62.0 C-6, 61.8 (app m,
), 54.9 C-2, 34.9 (d, J = 129.6 Hz, C(O)CH P(O)(OEt) ), 31.4 (t, J = 21.4
CF ), 20.7 C(O)CH , 20.61 C(O)CH , 20.59 C(O)CH , 16.21 (d, J = 4.7 Hz,
), 16.15 (d, J = 4.8 Hz, OCH ); F NMR (379 MHz, CDCl ) δ -81.30, -113.85, -
1
2
3
,
H-2), 3.89-3.78 (m, 2H, OCH
2
2
2
)
5
3
2
2
2
)
5
3
1
3
2
3
); C NMR
(
3
3
3
3
C(O)CH
2
2
2
)
5
3
6
2.8 (d, J = 6.6 Hz, OCH
OCH CH (CF CF
Hz, OCH CH (CF
OCH CH
2
3
2
3
2
2
2
)
5
3
2
2
2
2
2
)
5
3
3
3
3
1
9
2
3
2
CH
3
3
20
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3
1
1
22.56, -123.38, -124.10, -126.65; P NMR (121 MHz, CDCl
3
): δ 22.52 (s, 1P); MS (ESI) m/z
+
+
[
M+H] calcd for C26
H
34
F
13NO12P: 830.1611; found: 830.1584 m/z [M+Na] calcd for
C
26
H
33
F
13NNaO12P: 852.1430; found: 852.1404.
.12.3. 3,4,6-Tri-O-acetyl-2-deoxy-2-diethylphosphonoacetamido-1-
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10-eicosafluorononylmethoxy)-β-D-glucopyranoside 25
,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10-
4
(
3
eicosafluorononylmethoxy)-β-D-glucopyranoside 22 (0.065 g, 0.073 mmol) and P(OEt)
3
(0.03
) δ
H), 5.37 (dd, J = 10.8
Hz, J = 9.5 Hz, 1H, H-3), 5.15 (t, J = 9.4 Hz, 1H, H-4), 4.87 (d, J = 8.2 Hz, 1H, H-1), 4.27 (dd, J
4.8 Hz, J = 12.3 Hz, 1H, H-6a), 4.33-4.21 (m, 6H, H-6b, 2xOCH CH , H-2), 4.17-4.08 (m, 2H,
OCH (CF CF H), 3.69-3.65 (m, 1H, H-5), 2.77 (d, J = 21.1 Hz, 2H, CH P(O)), 2.11 (s, 3H,
OAc), 2.05 (s, 3H, OAc), 2.03 (s, 3H, OAc); C NMR (101 MHz, CDCl ) δ 171.2 C(O)CH
70.9 C(O)CH , 169.4 C(O)CH , 166.2 (d, J = 3.6 Hz, C(O)CH P(O)(OEt) ), 116.2-96.0 (m,
CF CF H), 100.4 C-1, 70.5 C-5, 68.3 C-3, 67.8 C-4, 62.0 (d, J = 6.6 Hz, OCH CH ), 62.1 (d, J
6.5 Hz, OCH CH ), 62.4 C-6, 60.5 (m, OCH (CF CF H), 52.6 C-2, 34.8 (d, J = 129.4 Hz,
P(O)(OEt) ), 20.9 C(O)CH , 20.5 C(O)CH , 20.4 C(O)CH , 16.21 (d, J = 4.8 Hz,
CH ), 16.18 (d, J = 4.6 Hz, OCH ); F NMR (376 MHz, CDCl
23.28, -129.22, -137.05; P NMR (162 MHz, CDCl
calcd for
20NNaO12P: 1020.1240; found: 1020.1248.
1
mL, 0.146 mmol) gave 25 as a white solid. Yield: (0.050 g) 68 %; H NMR (403MHz, CDCl
3
6
.86 (d, J = 9.2 Hz, 1H, NH), 6.07 (tt, J = 4.9 Hz, J = 51.7 Hz, 1H, CF
2
=
2
3
2
2
)
9
2
2
1
3
3
3
,
1
3
3
2
2
(
2
)
9
2
2
3
=
2
3
2
2
)
9
2
C(O)CH
2
2
3
3
3
1
9
OCH
2
3
2
CH
3
3
) δ -119.91, -121.88, -
3
1
+
1
3
): δ 22.18 (s, 1P); MS (ESI) m/z [M+H]
+
C
29
H
33
F20NO12P: 998.1421; found: 998.1423 m/z [M+Na]
calcd for
C
29
32
H F
21
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4
.13. General procedure for the synthesis of fluorinated sugar azides 26, 27, 28
Monosaccharides with 2-chloroacetamido group 20, 21, 22 (1 equiv.) were heated overnight with
KI (2.3 equiv.) and NaN (3 equiv.) in anhydrous DMF (12 mL for 1 mmol of sugar) under an
3
argon atmosphere in 80 °C. The solvent was evaporated from the reaction mixture and the
residue diluted with EtOAc (2 mL for 1 mmol of sugar) washed with brine (2x3 mL for 1 mmol
of sugar). The organic phase was dried over Na
2
SO and solvent was evaporated under reduced
4
pressure. The crude product was purified by column chromatography using a 3:1 mixture of
EtOAc/CH as eluent to afforded compounds 26, 27, 28.
4
.13.1. 3,4,6-Tri-O-acetyl-2-azidoacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6,7,7-
pentadecafluoromethoxy)-β-D-glucopyranoside 26
,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluoroethoxy)-
3
β-D-glucopyranoside 21 (0.032 g, 0.042 mmol), KI (0.016 g, 0.096 mmol) and NaN
3
(0.008 g,
) δ 6.74
1
0
.126 mmol) gave 26 as a white solid. Yield: (0.030 g) 93 %; H NMR (403 MHz, CDCl
3
(
d, J = 8.6 Hz, 1H, NH), 5.42 (dd, J = 9.2 Hz, J = 10.6 Hz, H, H-3), 5.09 (t, J = 9.6 Hz, 1H, H-4),
.95 (d, J = 8.2 Hz, 1H, H-1), 4.33-4.23 (m, 2H, H-6a, OCH (CF CF ), 4.17 (dd, J = 12.7 Hz, J
2.7 Hz, 1H, H6b), 3.94-3.83 (m, 3H, H-2, CH ), 3.77 (ddd, J = 2.4 Hz, J = 4.6 Hz, J = 9.9
Hz, 1H, H-5), 2.10 (s, 3H, OAc), 2.038 (s, 3H, OAc), 2.036 (s, 3H, OAc); C NMR (101 MHz,
CDCl ) δ 170.59 C(O)CH , 170.54 C(O)CH , 169.2 C(O)CH , 169.2 C(O)CH , 122.5-109.5
m, (CF CF ), 100.4 C-1, 72.4 C-5, 71.3 C-3, 68.3 C-4, 65.2 C-6, 65.3 (t, J = 65.3 Hz,
CF
4
2
2
)
6
3
=
2
N
3
1
3
3
3
3
3
2
N
3
(
2
)
6
3
1
9
CH
2
(CF
2
)
6
3
), 54.5 C-2, 52.5 CH
2
N
3
, 20.64 C(O)CH
3
, 20.57 C(O)CH3, 20.54 C(O)CH ; F
3
NMR (379 MHz, CDCl
3
) δ: -81.24, -120.38, -122.52, -123.22, -123.62, -126.60; MS (ESI) m/z
22
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+
+
[
M+H] calcd for C22
H
22
F
15
N
4
O
9
: 771.1147; found: 771.1155 m/z [M+Na] calcd for
: 793.0967; found: 793.0977.
.13.2. 3,4,6-Tri-O-acetyl-2-azidoacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6-
tridecafluoroethoxy)-β-D-glucopyranoside 27
,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluoroethoxy)-
C
22
H
21
F
15
N
4
NaO
9
4
3
β-D-glucopyranoside 21 (0.114 g, 0.157 mmol), KI (0.060 g, 0.361 mmol) and NaN
3
(0.031 g,
) δ 6.43
1
0
.471 mmol) gave 27 as a white solid. Yield: (0.108 g) 94 %; H NMR (403 MHz, CDCl
3
(
d, J = 8.7 Hz, 1H, NH), 5.36 (dd, J = 10.6 Hz, J = 9.3 Hz, 1H, H-3), 5.08 (t, J = 9.6 Hz, 1H, H-
), 4.80 (d, J = 8.3 Hz, 1H, H-1), 4.27 (dd, J = 12.3 Hz, J = 4.8 Hz, 1H, H-6a), 4.18-4.11 (m, 3H,
H-6b, OCH CH (CF CF ), 3.93 (s, 2H, CH ), 3.87-3.80 (m, 1H, H-2), 3.74 (ddd, J = 10.0
Hz, J = 4.9 Hz, J = 2.5 Hz, 1H, H-5), 2.52-2.34 (m, 2H, CH (CF CF ), 2.09 (s, 3H, OAc), 2.04
s, 3H, OAc), 2.03 (s, 3H, OAc); C NMR (101 MHz, CDCl ) δ 170.7 C(O)CH , 170.6
C(O)CH , 169.4 C(O)CH , 167.1 C(O)CH , 121.7-106.4 (m, (CF CF ), 100.5 C-1, 72.0 C-5,
1.6 C-3, 68.4 C-4, 61.9 C-6, OCH CH (CF CF , 54.7 C-2, 52.5 CH , 31.3 (t, J = 21.4 Hz,
), 20.6 C(O)CH , 20.59 C(O)CH , 20.57 C(O)CH
4
2
2
2
)
5
3
2
N
3
2
2
)
5
3
1
3
(
3
3
3
3
2
N
3
2
)
5
3
7
2
2
2
)
5
3
2
N
3
1
9
OCH
2
CH
2
(CF
2
)
5
CF
3
3
3
3
; F NMR (379 MHz,
+
CDCl
3
) δ -81.30, -113.91, -122.41, -123.38, -124.12, -126.64; MS (ESI) m/z [M+H] calcd for
+
C
22
H
24
F
13
N
4
O
9
: 735.1336; found: 735.1330 m/z [M+Na] calcd for C22
H
23
F
13
N
4
NaO : 757.1155;
9
found: 757.1148.
.13.3. 3,4,6-Tri-O-acetyl-2-azidoacetamido-2-deoxy-1-(1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10-
eicosafluorononylmethoxy)-β-D-glucopyranoside 28
,4,6-Tri-O-acetyl-2-chloroacetamido-2-deoxy-1-(1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10-
eicosafluorononylmethoxy)-β-D-glucopyranoside 22 (0.090 g, 0.101 mmol), KI (0.037 g, 0.225
4
3
23
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1
mmol) and NaN
NMR (403MHz, CDCl
CF H), 5.42 (dd, J = 10.6 Hz, J = 9.2 Hz, 1H, H-3), 5.11 (t, J = 9.4 Hz, 1H, H-4), 4.95 (d, J = 8.2
Hz, 1H, H-1), 4.34-4.25 (m, 3H, H-6a, OCH (CF CF H), 4.17 (dd, J = 12.4 Hz, J = 2.5 Hz, 1H,
H-6b), 3.96-3.84 (m, 3H, CH , H-2), 3.76 (ddd, J = 9.9 Hz, J = 4.9 Hz, J = 2.4 Hz, 1H, H-5),
.11 (s, 3H, OAc), 2.05 (s, 3H, OAc), 2.04 (s, 3H, OAc); C NMR (101 MHz, CDCl
C(O)CH , 170.9 C(O)CH , 169.5 C(O)CH , 169.5 C(O)CH , 116.2-98.0 (m, (CF
00.4 C-1, 70.5 C-5, 68.2 C-3, 67.7 C-4, 65.0 C-6, 61.5 CH (CF CF H, 53.6 C-2, 52.3 CH
; F NMR (379 MHz, CDCl ) δ -120.36, -122.40, -
: 903.1146; found:
3
(0.033 g, 0.505 mmol) gave 28 as a white solid. Yield: (0.081 g) 89 %; H
3
) δ 6.68 (d, J = 8.6 Hz, 1H, NH), 6.06 (tt, J = 5.1 Hz, J = 51.8 Hz, 1H,
2
2
2
)
9
2
2
N
3
1
3
2
3
) δ 171.2
CF H),
3
3
3
2
N
3
2
)
9
2
1
2
1
9
4
2
2
)
9
2
2
3
N ,
1
9
0.7 C(O)CH
3
, 20.5 C(O)CH
3
, 20.4 C(O)CH
3
3
+
23.76, -129.60, -137.41; MS (ESI) m/z [M+H] calcd for C25
H
23
F
20
N
4
O
9
+
03.1144; m/z [M+Na] calcd for C25
H
22
F
20
N
4
NaO : 925.0965; found: 925.0962.
9
.14. General procedure for the synthesis of fluorinated phosphoramidates 29, 30, 31
Corresponding florinated azides 26, 27, 28 in anhydrous DCM (43.3 mL for 1 mmol of sugar),
molecular sieves 4Å (0.213 g for 1 mmol of sugar) was added and the reaction mixture was
stirred for 1 h at room temperature. Next, P(OEt) (5 equiv.) was added and stirring was
3
continued overnight. Then, it was diluted with DCM, passed through a pad of Celite and solvent
was evaporated under reduced pressure. The crude product was purified by column
chromatography (DCM:MeOH 20:1, v/v) gave 29, 30, 31.
4
.14.1. 3,4,6-Tri-O-acetyl-2-deoxy-2-diethylphosphoramidoacetamido-1-
1,1,1,2,2,3,3,4,4,5,5,6,6,7,7-pentadecafluoromethoxy)-β-D-glucopyranoside 29
,4,6-Tri-O-acetyl-2-azidoacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6,7,7-
pentadecafluoromethoxy)-β-D-glucopyranoside 26 (0.051 g, 0.066 mmol) and P(OEt)
(
3
3
(0.06 mL,
24
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1
0
.330 mmol) gave 29 as a white solid. Yield: (0.044 g) 76%; H NMR (400 MHz, CDCl
d, J = 8.6 Hz, 1H, NH), 5.42 (dd, J = 10.4 Hz, J = 9.3 Hz, 1H, H-3), 5.08 (t, J = 9.6 Hz, 1H, H-
), 4.99 (d, 1H, J = 8.2 Hz, H-1), 4.28 (dd, J = 12.3 Hz, J = 4.8 Hz, 1H, H-6a), 4.16 (dd, J = 2.4
Hz, J = 12.3 Hz, 1H, H-6b), 4.12-4.03 (m, 6H, 2xCH CH , OCH (CF CF ), 3.94-3.84 (m, 1H,
H-2), 3.76 (ddd, J = 9.9 Hz, J = 5.0 Hz, J = 2.3 Hz, 1H, H-5), 3.53-3.99 (m, 3H, CH NHP(O)),
3
) δ 7.25
(
4
2
3
2
2
)
6
3
2
1
3
2
.09 (s, 3H, OAc), 2.03 (s, 3H, OAc), 2.02 (s, 3H, OAc), 1.36-1.28 (m, 6H, 2xCH
151 MHz, CDCl ) δ 170.6 C(O)CH , 170.5 C(O)CH , 169.5 C(O)CH , 169.4 (d, J = 9.4 Hz,
NHP(O)(OEt) ), 122.8-108.6 (m, (CF CF ), 100.6 C-1, 72.3 C-5, 71.7 C-3, 68.7 C-4,
CH ), 62.9 (d, J = 6.6 Hz, OCH CH ), 62.1 C-6, 64.5 (m,
), 54.5 C-2, 44.8 (s, C(O)CH NHP(O)(OEt) ), 20.6 C(O)CH , 20.5 C(O)CH
, 16.09 (d, J = 6.8 Hz, OCH CH ), 16.06 (d, J = 6.7 Hz, OCH
3
); C NMR
(
3
3
3
3
C(O)CH
2
2
2
)
5
3
6
3.0 (d, J = 6.6 Hz, OCH
2
3
2
3
OCH
2
(CF
2
)
6
CF
3
2
2
3
3
,
1
9
2
0.3 C(O)CH
3
2
3
2
CH ); F NMR
3
3
1
(
282 MHz, CDCl
3
) δ -81.29, -120.68, -122.54, -123.27, -123.80, -126.65; P NMR (121 MHz,
+
CDCl
3
): δ 8.51 (s, 1P); MS (ESI) m/z [M+H] calcd for C26
H
33
F
15
2
N O12P: 881.1532; found:
+
8
4
81.1555 m/z [M+Na] calcd for C26
H
32
F
15
N NaO12P: 903.1351; found: 903.1372.
2
.14.2. 3,4,6-Tri-O-acetyl-2-deoxy-2-diethylphosphoramidoacetamido-1-
(
1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluoroethoxy)-β-D-glucopyranoside 30
,4,6-Tri-O-acetyl-2-azidoacetamido-2-deoxy-1-(1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluoroethoxy)-
3
β-D-glucopyranoside 27 (0.102 g, 0.139 mmol) and P(OEt)
3
(0.12 mL, 0.695 mmol) gave 30 as a
) δ 7.18 (d, J = 8.8 Hz, 1H, NH),
1
white solid. Yield: (0.100 g) 85%; H NMR (403 MHz, CDCl
3
5
1
6
.34 (dd, J = 10.3 Hz, J = 9.4 Hz, 1H, H-3), 5.06 (t, J = 9.6 Hz, 1H, H-4), 4.83 (d, J = 8.3 Hz,
H, H-1), 4.26 (dd, J = 12.3 Hz, J = 5.1 Hz, 1H, H-6a), 4.15 (dd, J = 2.4 Hz, J = 12.3 Hz, 1H, H-
b), 4.12-4.03 (m, 4H, 2xCH CH ), 3.92-3.82 (m, 3H, H-2, OCH CH (CF CF ), 3.73 (ddd, J =
2
3
2
2
2
)
5
3
25
ACCEPTED MANUSCRIPT

ACCEPTED MANUSCRIPT
9
.9 Hz, J = 5.0 Hz, J = 2.3 Hz, 1H, H-5), 3.58-3.43 (m, 3H, CH
2
NHP(O)), 2.52-2.36 (m, 2H,
), 2.08 (s, 3H, OAc), 2.02 (s, 3H, OAc), 2.01 (s, 3H, OAc), 1.36-1.29 (m, 6H,
) δ 170.7 C(O)CH , 170.6 C(O)CH , 170.5 C(O)CH , 169.4
NHP(O)(OEt) ), 120.8-107.6 (m, (CF CF ), 100.6 C-1, 72.2 C-5, 71.9
C-3, 68.6 C-4, 62.95 (d, J = 6.6 Hz, OCH CH ), 62.90 (d, J = 6.5 Hz, OCH CH ), 62.0 C-6, 61.6
appm, OCH CH (CF CF ), 54.4 C-2, 44.8 (s, C(O)CH NHP(O)(OEt) ), 31.4 (t, J = 21.7 Hz,
(CF CF ), 20.64 C(O)CH , 20.61 C(O)CH , 20.57 C(O)CH , 16.09 (d, J = 6.8 Hz,
), 16.04 (d, J = 6.8 Hz, OCH ); F NMR (282 MHz, CDCl ) δ -81.29, -113.99, -
CH
2
(CF
2
)
5
CF
3
1
3
2
xCH
3
); C NMR (101 MHz, CDCl
3
3
3
3
(
d, J = 9.4 Hz, C(O)CH
2
2
2
)
5
3
2
3
2
3
(
2
2
2
)
5
3
2
2
OCH
2
2
CH
2
3
2
)
5
3
3
3
3
1
9
OCH
CH
2
CH
3
3
3
1
1
22.42, -123.40, -124.15, -126.66; P NMR (121 MHz, CDCl
3
): δ 8.59 (s, 1P); MS (ESI) m/z
+
+
[
M+H] calcd for C26
NaO12P: 867.1539; found: 867.1530.
.14.3. 3,4,6-Tri-O-acetyl-2-deoxy-2-diethylphosphoramidoacetamido-
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10-eicosafluorononylmethoxy)-β-D-glucopyranoside 31
,4,6-Tri-O-acetyl-2-azidoacetamido-2-deoxy-1-(1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10-
H
35
F
13
N
2
O
12P: 845.1720; found: 845.1706 m/z [M+Na] calcd for
C
26
H
34
F
13
N
2
4
(
3
eicosafluorononylmethoxy)-β-D-glucopyranoside 28 (0.052 g, 0.058 mmol) and P(OEt)
3
(0.05
) δ
1
mL, 0.290 mmol) gave 30 as a white solid. Yield: (0.053 g) 92%; H NMR (400MHz, CDCl
3
6
.65 (d, J = 8.6 Hz, 1H, NH), 6.06 (tt, J = 5.1 Hz, J = 51.8 Hz, CF
.2 Hz, 1H, H-3), 5.10 (t, J = 9.4 Hz, 1H, H-4), 4.96 (d, J = 8.2 Hz, 1H, H-1), 4.34-4.23 (m, 7H,
CH , OCH (CF CF H), 4.17 (dd, J = 12.4 Hz, J = 2.5 Hz, 1H, H-6b), 3.96-3.84
m, 3H, H-2, CH NHP(O)), 3.77 (ddd, J = 9.9 Hz, J = 4.9 Hz, J = 2.4 Hz, 1H, H-5), 2.10 (s, 3H,
OAc), 2.05 (s, 3H, OAc), 2.03 (s, 3H, OAc); F NMR (379 MHz, CDCl
2
H), 5.44 (dd, J = 10.6 Hz, J =
9
H-6a, 2xCH
2
3
2
2
)
9
2
(
2
1
9
3
) δ -120.36, -122.42, -
26
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+
1
1
4
23.74, -129.72, -137.39; MS (ESI) m/z [M+H] calcd for C29
H
34
F
20
N
2
O
12P: 1013.1530; found:
NaO12P: 1035.1349; found: 1035.1349.
.15. General synthesis of phosphonates 32 and 34
+
013.1529 m/z [M+Na] calcd for C29
H
33
F
20
N
2
To a solution of sugar chloride 19 (1 equiv.) or sugar bromide 33 (1 equiv.) in MeCN (30 mL for
mmol of sugar), KI (2.26 equiv.) and P(OEt) (2 equiv.) were added. The reaction mixture was
1
3
heated in 65 °C for 3.5-4 h. The solvent was evaporated and the crude product was purified by
column chromatography using a 20:1 mixture of DCM/MeOH as eluent.
4
1
.15.1. 2-Diethylhosphonoacetamido-1,2,4,6-tetra-O-acetyl-2-deoxy-β-D-glucopyranoside 32
,3,4,6-tetra-O-acetyl-2-chloroacetamido-2-deoxy-β-D-glucopyranoside 19 (0.021 g, 0.050
1
mmol) and P(OEt)
3
(0.02 mL, 0.100 mmol) gave 32 as a white solid. Yield: (0.018 g) 69%; H
NMR (600 MHz, CDCl
3
): δ 6.81 (d, J = 9.0 Hz, 1H, NH), 5.85 (d, J = 8.8 Hz, 1H, H-1), 5.33
(
dd, J = 9.3 Hz, J = 10.6 Hz, 1H, H-4), 5.10 (t, J = 9.6 Hz, 1H, H-3), 4.29 (dd, J = 4.5 Hz, J =
1
2.5 Hz, 1H, H-6a), 4.20-4.14 (m, 1H, H-2), 4.13-4.09 (m, 5H, H-6b, 2xOCH
2.2 Hz, J = 4.5 Hz, J = 10.0 Hz, 1H, H-5), 2.81 (d, J = 21.0 Hz, 2H, CH P(O)(OEt)
H, OAc), 2.09 (s, 3H, OAc), 2.05 (s, 3H, OAc), 2.03 (s, 3H, OAc), 1.37-1.32 (m, 6H, 2xCH
2
CH
3
), 3.87 (ddd, J
), 2.13 (s,
);
, 169.3
), 91.9 C-1, 72.6 C-5, 72.0 C-3, 68.2 C-4,
), 61.6 C-6, 53.3 C-2, 35.3 (d, J = 129.9 Hz, C(O)CH P(O)(OEt) ),
=
2
2
3
3
1
3
C NMR (101 MHz, CDCl
C(O)CH , 164.5 (d, J = 4.1 Hz, C(O)CH
2.9 (d, J = 6.4 Hz, OCH CH
0.9 C(O)CH , 20.71 C(O)CH
3
): δ 170.7 C(O)CH
3
, 170.6 C(O)CH
3
, 169.4 C(O)CH
3
3
2
P(O)(OEt)
2
6
2
3
2
2
2
3
3
, 20.68 C(O)CH
3
, 20.60 C(O)CH
3
, 16.28 (d, J = 6.1 Hz,
3
1
+
OCH
2
CH
3
); P NMR (243 MHz, CDCl
3
): δ 21.53 (s, 1P); MS (ESI) m/z [M+H] calcd for
+
C
20
H
33NO13P: 526.1690; found: 526.1698 m/z [M+Na] calcd for C20
H
32NNaO13P: 548.1509;
found: 548.1522.
27
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4
1
0
.15.2. 2-Diethylphosphonopropanamido-1,2,4,6-tetra-O-acetyl-2-deoxy-β-D-glucopyranoside 34
,3,4,6-tetra-O-acetyl-2-(3-bromopropionylamido)-2-deoxy- β-D-glucopyranoside (33) (0.097 g,
.200 mmol) and P(OEt)
3
(0.1 mL, 0.600 mmol) gave 34 as a white solid. Yield: (0.046 g) 43
): δ 7.07 (d, J = 9.2 Hz, 1H, NH), 5.76 (d, J = 8.8 Hz, 1H, H-1),
1
%
; H NMR (400 MHz, CDCl
3
5
4
2
2
.23 (appt, J = 9.9 Hz, 1H, H-3), 5.10 (t, J = 9.6 Hz, 1H, H-4), 4.30-4.25 (m, 2H, H-6a, H-6b),
.14-4.05 (m, 5H, 2xOCH CH , H-2), 3.81 (ddd, J = 2.2 Hz, J = 4.6 Hz, J = 9.8 Hz, 1H, H-5),
.35-2.45 (m, 2H, CH CH P(O)(OEt) ), 2.06-1.96 (m, 2H, CH P(O)(OEt) ), 2.10 (s, 3H, OAc),
.08 (s, 3H, OAc), 2.02 (s, 6H, 2xOAc), 1.37-1.26 (m, 6H, 2xCH
): δ 171.4 C(O)CH , 171.3 C(O)CH , 170.72 C(O)CH , 170.68 C(O)CH
Hz, C(O)CH CH P(O)(OEt) ), 92.4 C-1, 72.66 C-5, 72.65 C-3, 68.1 C-4, 62.05 (d, J = 6.5 Hz,
OCH CH ), 61.99 (d, J = 6.6 Hz, OCH CH ), 61.7 C-6, 53.0 C-2, 29.1 (d, J = 3.6 Hz,
, 20.72 C(O)CH , 20.74 (d, J = 143.8 Hz,
2
3
2
2
2
2
2
1
3
3
); C NMR (151 MHz,
CDCl
3
3
3
3
3
, 169.4 (d, J = 4.4
2
2
2
2
3
2
3
C(O)CH
C(O)CH
2
CH
2
2
P(O)(OEt)
P(O)(OEt)
2
), 20.9 C(O)CH
), 20.65 C(O)CH , 20.59 C(O)CH
3
3
2
CH
2
3
3
, 16.4 (d, J = 5.6 Hz, 2xOCH
2
CH );
3
3
1
+
P NMR (121 MHz, CDCl
40.1846; found: 540.1852 m/z [M+Na] calcd for C21
.16. 1,3,4,6-tetra-O-acetyl-2-(3-bromopropionylamido)-2-deoxy-β-D-glucopyranoside 33
3
): δ 31.33 (s, 1P); MS (ESI) m/z [M+H] calcd for C21
H
35NO13P:
+
5
4
H34NNaO13P: 562.1665; found: 562.1674.
A solution of 2-deoxy-2-amino-1,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl hydrochloride (0.099
g, 0.258 mmol) in anhydrous DCM (3.3 mL) was cooled to 0 °C. Next Et N (0.1 mL) was added
3
followed by DCC (0.186 g, 0.903 mmol) and 3-bromopropionic acid (0.276 g, 1.806 mmol). The
reaction mixture was allowed to warm up to room temperature and stirred overnight. After
finishing the precipitate was filtered and the solvents were evaporated. Product was purified by
1
column chromatography (EtOAc:CH 1:1, v/v) gave 33 as white solid. Yield: (0.092 g) 75%; H
28
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NMR (403 MHz, CDCl
3
): δ 6.02 (d, J = 9.4 Hz, 1H, NH), 5.72 (d, J = 8.8 Hz, 1H, H-1), 5.21
(
dd, J = 9.4 Hz, J = 10.5 Hz, 1H, H-3), 5.13 (t, J = 9.6 Hz, 1H, H-4), 4.41-4.32 (m, 1H, H-2),
4
.28 (dd, J = 4.7 Hz, J = 12.5 Hz, 1H, H-6a), 4.13 (dd, J = 2.6 Hz, J = 12.3 Hz, 1H, H-6b), 3.85
Br), 2.70 (t, J =
Br), 2.13 (s, 3H, OAc), 2.10 (s, 3H, OAc), 2.06 (s, 3H, OAc), 2.05 (s, 3H,
): δ 171.3 C(O)CH , 170.7 C(O)CH , 169.7 C(O)CH , 169.6
Br, 92.4 C-1, 72.8 C-5, 72.4 C-3, 67.9 C-4, 61.6 C-6, 52.8 C-2,
(
ddd, J = 2.2 Hz, J = 4.6 Hz, J = 9.8 Hz, 1H, H-5), 3.61 (t, J = 6.2 Hz, 1H, CH
2
6
.2 Hz, 1H, CH
2
CH
2
1
3
OAc); C NMR (101 MHz, CDCl
C(O)CH , 169.3 C(O)CH CH
9.6 C(O)CH CH Br, 27.3 CH
3
3
3
3
3
2
2
3
2
2
2
CH
2
Br, 21.0 C(O)CH
3
, 20.9 C(O)CH
3
, 20.7 C(O)CH , 20.6
3
+
C(O)CH
3
; MS (ESI) m/z [M+H] calcd for C17
H25BrNO10: 482.0662; found: 482.0676 m/z
+
[
M+Na] calcd for C17
H24BrNNaO10: 504.0481; found: 504.0485.
4
.17. General synthesis of phosphoramidates 36 and 38
To the mixture of 35 (1 equiv.) or 37 (1 equiv.) in anhydrous DCM (43.3 mL for 1 mmol of
sugar), molecular sieves 4Å (0.213 g for 1 mmol of sugar) was added and the reaction mixture
was stirred for 1h at room temperature. Next, P(OEt) (5 equiv.) was added and stirring was
3
continued overnight. Then, it was diluted with DCM, passed through a pad of Celite and solvent
was evaporated under reduced pressure. The crude product was purified by column
chromatography (DCM:MeOH 20:1, v/v) gave 36 or 38.
4
1
.17.1. 2-diethylphosphoramidoacetamido-1,2,4,6-tetra-O-acetyl-2-deoxy-β-D-glucopyranose 36
,3,4,6-tetra-O-acetyl-2-azidoacetamide-2-deoxy-β-D-glucopyranose 35 (0.079 g, 0.184 mmol)
1
and P(OEt)
3
(0.16 mL, 0.920 mmol) gave 36 as a white solid. Yield: (0.068 g) 69%; H NMR
(
400 MHz, CDCl
3
): δ 7.28 (d, J = 9.5 Hz, 1H, NH), 5.86 (d, J = 8.7 Hz, 1H, H-1), 5.38 (dd, J =
9
.4 Hz, J = 10.5 Hz, 1H, H-3), 5.13 (t, J = 9.7 Hz, 1H, H-4), 4.35-4.25 (m, 1H, H-6a, H-2), 4.16-
29
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