Synthesis of cluster mannosides carrying a photolabile diazirine group

Article abstract of DOI:10.1002/ejoc.200500692

We are investigating the mechanisms underlying the carbohydrate-specific adhesion of bacteria such as Escherichia coli to the glycocalyx of their potential host cells. E. coli possess protein appendages, which are called type 1 fimbriae. Part of type 1 fimbriae is a protein named FimH, which is a mannose-specific lectin. We wish to use photoaffinity labeling to elucidate mannose binding sites on FimH. Thus we report the synthesis of di- and trivalent cluster mannosides, which carry a photolabile diazirine group. The diazirine group was introduced by a convergent approach using thiourea bridging (products 6, 13, 17, and 27) or in a divergent synthesis leading to the divalent cluster mannoside 31. Wiley-VCH Verlag GmbH & Co. KGaA, 2006.

Full text of DOI:10.1002/ejoc.200500692

FULL PAPER
DOI: 10.1002/ejoc.200500692
Synthesis of Cluster Mannosides Carrying a Photolabile Diazirine Group
Mark Walter,[a] Michaela Wiegand,[a] and Thisbe K. Lindhorst*[a]
Keywords: Carbohydrates / Glycoclusters / Photoaffinity labeling / Diazirines / Thiourea bridging
We are investigating the mechanisms underlying the carbo-
hydrate-specific adhesion of bacteria such as Escherichia coli
to the glycocalyx of their potential host cells. E. coli possess
protein appendages, which are called type 1 fimbriae. Part
of type 1 fimbriae is a protein named FimH, which is a man-
nose-specific lectin. We wish to use photoaffinity labeling to
elucidate mannose binding sites on FimH. Thus we report
the synthesis of di- and trivalent cluster mannosides, which
carry a photolabile diazirine group. The diazirine group was
introduced by a convergent approach using thiourea bridg-
ing (products 6, 13, 17, and 27) or in a divergent synthesis
leading to the divalent cluster mannoside 31.
(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim,
Germany, 2006)
To verifiy this hypothesis using the photoaffinity labeling
technology, we have defined di- and trivalent cluster man-
nosides as our target molecules, which carry a photolabile
diazirine group. Ideally, incubation of such photolabile clus-
ter mannosides with FimH leads to covalently linked gly-
cocluster-FimH conjugates, giving information about
carbohydrate binding sites on the protein. Thus, the synthe-
sis of divalent and trivalent photolabeled cluster mannos-
ides is described in this contribution.
Introduction
Photoaffinity labeling is a powerful method of biological
chemistry[1] to study receptor binding sites, such as the
active site of an enzyme, e.g., or the carbohydrate recogni-
tion domain (CRD) of a lectin, respectively. It enables di-
rect probing of a target receptor through a covalent bond,
which is photochemically introduced between the ligand
and the specific protein.
Among several photoreactive groups commonly used in
photoaffinity labeling,[2] diazirines[3] have often been shown
to be advantageous over others.[4] Upon irradiation diazir-
ines produce a reactive carbene after the light-induced loss
of nitrogen. This photogenerated intermediate is very reac-
tive and can insert into single bonds allowing the charac-
terization of carbohydrate-binding proteins, for instance.[5]
In the course of our work on the investigation of carbo-
hydrate binding to the bacterial lectin FimH[6] we have en-
visaged photoaffinity labeling of FimH using suitable
carbohydrate derivatives which carry a photolabile diazirine
group. The bacterial lectin FimH is part of the so-called
type 1 fimbriae, which are protein appendages on the sur-
face of Escherichia coli and are utilized for carbohydrate-
specific adhesion of the bacteria to the host cell glycocalyx.
X-ray studies have revealed that the lectin domain of type
1 fimbriae, the protein FimH, possesses a CRD at its tip,
which can accommodate one α-mannoside residue.[7] How-
ever, theoretical studies[8] as well as testing results with
multivalent mannosyl clusters[9] have suggested, that there
might be additional carbohydrate binding domains on
FimH.
Results and Discussion
We have recently reported a convergent approach for the
synthesis of diazirine-labeled carbohydrate derivatives,
which is based on thiourea bridging of an amino-function-
alized diazirine derivative (4, Scheme 1) and isothiocyanato-
functionalized sugars.[10] As thiourea-bridged glycoclusters
have been shown to be highly interesting probes for bio-
logical studies,[11] we have also envisaged this approach for
the preparation of photolabeled glycoclusters. Earlier on,
thiourea-bridging has led to photolabile glycoclusters carry-
ing three diazirine groups utilizing the diisothiocyanato-
functionalized mannose derivative 1.[10] However, we fig-
ured that glycoclusters carrying more than one diazirine
group are not ideally suited as photoaffinity probes for
FimH because irradation would always activate all photola-
bile groups present in the molecule and eventually lead to
a puzzling armada of products. Therefore, we tried to syn-
thesise mannose clusters with a single diazirine group and
have attempted to use an anlogous approach as reported
earlier.[10] Thus, the isothiocyanato-functionalized manno-
syl isothiocyanate 1 was treated with tris(2-aminoethyl)-
amine (2) to obtain the glycocluster 3 in 70% yield
(Scheme 1). Then, we have evaluated the feasibility of
mono-functionalization of 3 with the diazirine 4, however,
only the synthesis of the tri-functionalized 5 was satisfac-
[a] Otto Diels Institute of Organic Chemistry, Christiana Albertina
University of Kiel,
24098 Kiel, Germany
Fax: +49-431-880-7410
E-mail: tklind@oc.uni-kiel.de
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M. Walter, M. Wiegand, T. K. Lindhorst
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Scheme 1. a) Dichloromethane, 0 °C, 2 h, 70%; b) dichloromethane, DIPEA, room temp., 2 h, 52%; c) 1 m NaOMe in MeOH, 0 °C, 1 h,
69%.
tory. While deprotection of 5 according to Zemplén[12] lead- One possibility is to turn the azide group into an NCS func-
ing to the water-soluble unprotected glycocluster 6 was con- tion in a Staudinger-type reaction by using triethyl phos-
venient, this was not the type of product we had targeted. phite and carbon disulfide[15] to yield the isothiocyanate 11.
To achieve the preparation of cluster mannosides with only The latter was allowed to react with the hydrochloride 4,
one diazirine group present in the molecule, we have em- which had to be converted into the respective amine by
ployed 2-hydroxymethyl-1,3-propandiol (Scheme 2) as the treatment with DIPEA, to finally furnish the targeted pho-
core molecule instead of the branched trisamine 2.
tolabeled bis-mannoside 12. Because the Staudinger reac-
A straightforward reaction sequence starting with 2-hy- tion as well as thiourea bridging in the following step gave
droxymethyl-1,3-propandiol led to the isopropylidene-pro- lower yields than usual, we assume that the reactivity at the
tected azide 7 via isopropylidination, tosylation of the re- focal point of 10 and 11 is diminished, possibly due to steric
maining hydroxy group and nucleophilic substitution by so- hindrance. Deprotection of 12 led to the OH-free com-
dium azide in one pot (Scheme 2). The azide 7 was purified pound 13 which can be regarded as a photolabile disaccha-
and subjected to cleavage of the dioxane ring to yield the ride mimetic.[16]
diol 8, which served as mannosyl acceptor in the following
For a second synthetic pathway we planned to reduce the
glycosylation reaction. Mannosylation was carried out ac- azide 10 to the respective amine 14. Surprisingly this was
cording to the trichloroacetimidate method[13] using the not possible although several different reaction conditions
benzoyl-protected mannosyl donor 9[14] and TMS-triflate as were applied. Finally, Staudinger reaction using triphenyl
the Lewis acid. Almost one equivalent of the glycosyl donor phosphane provided the desired product 14 in moderate
per hydroxy group was enough to deliver the target com- yield, which was used without further purification
pound 10 in 75% yield.
(Scheme 2). The reaction with the NCS-functionalized
The azido group at the focal point of the divalent man- mannose derivative 15[10] carrying a diazirine group led to
noside 10 offers several options for further derivatization. the trivalent mannosyl cluster 16, which could be de-acyl-
720
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Eur. J. Org. Chem. 2006, 719–728

Synthesis of Cluster Mannosides Carrying a Photolabile Diazirine Group
FULL PAPER
Scheme 2. a) DMP, acetone, TsOH, THF, room temp., 1.5 h; then b) TsCl, pyridine, room temp., 12 h; then c) NaN3, TBABr, toluene,
60 °C, 24 h, 41% over three steps. d) HCl, MeOH, room temp., 1 h, quant. e) TMS-OTf, dichloromethane, Ar, 0 °C Ǟ room temp., 1 d,
75%. f) CS2, P(OEt)3, toluene, 80 °C, 8 h, 56%. g) PPh3, THF, Ar, 0 °C Ǟ room temp., 1 h; then h) H2O, 23% over two steps. i) 4,
DIPEA, dichloromethane, room temp., 2 h, 48%. k) 1 m NaOMe in MeOH/THF, room temp., 1 h, 65% for 13; 63% for 17.
ated according to Zemplén to yield 17. Both unprotected glycosylation. Again, only a minimum excess of the manno-
glycoclusters 13 and 17 carry differently positioned single syl donor 9 was necessary to deliver the trivalent cluster
diazirine groups. This is a favorable variation with regard mannoside 24 in good yield.
to the eventual biological application of these molecules, as
The azido-functionalized cluster 24 could be converted
the optimal position of the photoactive group within the into its isothiocyanato-functionalized analogue 25, as de-
carbohydrate probe is not known and can hardly be antici- scribed for 10 and thiourea bridging with 4 delivered the
pated.
photolabeled trivalent cluster mannoside 26. Unfortunately,
A similar approach as applied for the synthesis of 13 was the yield of the following deprotection reaction to give 27
then used to prepare a branched trivalent cluster mannoside did not exceed 55%.
based on a pentaerythritol derivative as the core molecule,
Finally we wanted to find out if it is possible to bind a
carrying the diazirine group on the scaffold (Scheme 3). The diazirine function to the focal point of glycodendrons of
synthesis started with triallyl pentaerythritol (18) which was the glycoglycerol type, which we have recently intro-
easily tosylated to 19. The tosylate was subjected to re- duced.[17] Therefore, we used the alkene 28, carrying two
ductive ozonization to yield the tris(hydroxyethyl) derivative isopropylidene-protected α-mannosyl residues as the start-
20. To facilitate purification, this product was acetylated to ing material. Ozonolysis was successful and work-up with
21 prior to the following nucleophilic substitution step with triphenylphosphane yielded the cluster mannoside 29 with
sodium azide, leading to the azido-functionalized, O-acetyl- a carbonyl function at its focal point. The ketone 29 was
protected triol 22, which could readily be purified. After subjected to the usual reaction sequence for the conversion
Zemplén deprotection the core molecule 23 was ready for into a diazirine ring.[18] Reaction with hydroxylamine-O-
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M. Walter, M. Wiegand, T. K. Lindhorst
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Scheme 3. a) TsCl, pyridine, room temp., 16 h, 81%. b) O3, MeOH, –78 °C, 0.5 h; then NaBH4, 46%. c) Ac2O, pyridine, room temp.,
16 h, 84%. d) NaN3, TBABr, DMF, 80 °C, 6 d, 93%. e) 1 m NaOMe in MeOH, room temp., 0.5 h, 91%. f) 9, TMS-OTf, dichloromethane,
Ar, 0 °C Ǟ room temp., 1 d, 67%. g) CS2, P(OEt)3, toluene, 80 °C, 8 h, 83%. h) 4, DIPEA, dichloromethane, 30 °C, 8 h, 80%. i) 1 m
NaOMe in MeOH/THF, room temp., 1 h, 55%.
sulfonic acid in liquid ammonia, followed by oxidation of out in MeOH or water and the respective insertion products
the intermediate diaziridine with iodine provided the photo- could be detected in the ESI-MS. However, when the herein
labeled divalent cluster mannoside 30 in acceptable yield. reported cluster mannosides were irradiated in the presence
Interestingly, a byproduct was formed and isolated in 24% of more complex compounds such as a model peptide, com-
yield, which was identified as the dimethoxy acetale ana- plex mixtures were obtained, also including unsaturated
logue of ketone 29 (Scheme 4). Possibly the ammonium ion, products due to undesired H-abstraction in the intermedi-
which is formed after the addition of hydroxylamine-O-sul- ate carbene. Additional irradiation studies with model pep-
fonic acid under the almost water-free reaction conditions tides as well as FimH are currently under investigation in
is acidic enough to catalyze the side reaction leading to the our laboratory.
observed byproduct.
Table 1. UV spectroscopic data of selected diazirines.
The acid-catalyzed deprotection of 30 was a high yielding
reaction leading to the diazirine-labeled cluster 31 with in-
tact glycosidic bonds.
A selection of the prepared diazirine-labeled saccharides
was irradiated at wavelengths of Ն 320 nm. Prior to irradia-
tion the UV spectra were recorded and the decrease of the
absorption maximum λmax of the individual compound was
Solvent Conc. [mM] λmax [nm] λmax disappearing after [min][a]
6
MeOH
0.5
0.3
0.3
0.2
342.2
341.1
342.2
333.3
30
60
60
13 H2O
17 H2O
27 H2O
decomposition
[a] After irradiation at λ Ն 320 nm; during irradiation UV spectra
observed during irradiation (Table 1). Irradition was carried were recorded in 5 minutes intervals.
722 Eur. J. Org. Chem. 2006, 719–728
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© 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Synthesis of Cluster Mannosides Carrying a Photolabile Diazirine Group
FULL PAPER
Scheme 4. a) O3, MeOH/dichloromethane, –78 °C, 20 min; then PPh3, 92%. b) NH3, H2NOSO3H, MeOH/dichloromethane, –50 °C Ǟ
–20 °C, 16 h; then 10% I2/MeOH, TEA, 0 °C, 49%. c) TFA /H2O, room temp., 10 min, 93%.
as the matrix. Ionisation was effected with a nitrogen laser at
337 nm. ESI mass spectra were measured with an Applied Biosys-
Conclusion
The reaction of photolabeled mannosyl clusters is feas-
ible according to a convergent as well as a divergent syn-
thetic approach. Strategies from various areas of organic
chemistry such as glycodendron-like growth, thiourea-
bridging and glycosidation can be favorably combined to
furnish molecules in which the photolabile group is dif-
ferently positioned. This is advantageous as the location of
the photolabel in the probe, which is ideal for photoaffinity
labeling of the protein under investigation, -FimH-, is not
known and can hardly be predicted. A series of structurally
varied, unprotected diazirine-labeled mannosyl clusters
were obtained by the described method. These compounds
lose nitrogen after 30 to 60 minutes upon irradiation at λ Ն
320 nm and will be evaluated for the investigation of the
FimH protein as well as other mannose-specific lectins.
tems Mariner ESI-TOF 5280.
Tris{2-[3Ј-(2ЈЈ,3ЈЈ,4ЈЈ-tri-O-acetyl-6ЈЈ-deoxy-6ЈЈ-isothiocyanato-α-D-
mannopyranosyl)thioureido]ethyl}amine (3): The diisothiocyanate 1
(155 mg, 0.40 mmol) was dissolved in dry dichloromethane (2 mL)
and cooled to 0 °C. A solution of tris(2-aminoethyl)amine (2,
19.6 µL, 19.3 mg, 0.132 mmol, 0.33 equiv.) in dry dichloromethane
(500 µL) was added in two subsequent portions after 30 minutes
intervals. After 1 h at 0 °C, stirring was continued for 1 h at room
temp., then the solvent was removed in vacuo and the residual
syrup purified by flash chromatography (cyclohexane/ethyl acetate,
9:1). The title compound was obtained as an amorphous powder
(123 mg, 94 µmol, 70%). Rf = 0.21 (ethyl acetate). [α]D = +28.3 (c
1
= 1.0 in CHCl3). H NMR (500 MHz, [D6]acetone): δ = 8.14 (br.
s, 3 H, 3 NHman), 7.53 (br. s, 3 H, 3 NHCH2), 6.18 (br. s, 3 H, 3
H-1), 5.51–5.43 (m, 3 H, 3 H-3), 5.36–5.26 (m, 6 H, 3 H-2, 3 H-4),
4.04–3.95 (m, 3 H, 3 H-5), 3.89 (dd, 3 H, 3 H-6Ј), 3.84–3.73 (m, 6
H, 3 H-6, 3 NHCHH), 3.72–3.60 (m, 3 H, 3 NHCHH), 2.84 [br. t,
6 H, 3 N(CH2)3], 2.16, 2.07, 1.99 [each s, each 9 H, 9 C(O)
3
3
3
3
2
Experimental Section
CH3] ppm; J2,3 = 2.6, J3,4 = 9.4, J5,6 = 3.9, J5,6Ј = 3.3, J6,6Ј
=
15.6 Hz. 13C NMR (125.75 MHz, [D6]acetone): δ = 184.26 (3 C=S),
170.25, 170.20, 170.18 (9 C=O), 132.59 (3 NCS), 80.75 (3 C-1),
70.58 (3 C-5), 70.02 (3 C-2), 69.58 (3 C-3), 67.27 (3 C-4), 53.47 [3
N(CH2)3], 46.40 (3 C-6), 43.94 (3 NHCH2), 20.80, 20.67, 20.59 [9
C(O)CH3] ppm. MALDI-TOF MS (DHB): m/z = 1310.9 [M]+
(calcd. m/z = 1310.27) for C48H66N10O21S6.
General Remarks: Optical rotations were measured with a Perkin–
Elmer polarimeter (20 °C, 589 nm, length of cuvette: 1 dm). Reac-
tions were monitored by TLC on silica gel GF254 (Merck) with
detection under UV and by charring with 10% sulfuric acid in eth-
anol and subsequent heating. Flash column chromatography was
performed on silica gel 60 (40–63 µm, Merck). NMR spectra were
recorded on Bruker AMX 400 or Bruker DRX 500 instruments.
Chemical shifts are relative to TMS or the solvent peaks of CDCl3
(δ = 7.24 ppm 1H, 77.0 ppm 13C) or [D6]acetone (δ = 2.04 ppm 1H,
29.3 ppm 13C). Where necessary, assignments were based on COSY
and HSQC experiments. Assignments indexed with an asterisk (*)
are interchangable. IR spectra were taken with a Perkin–Elmer FT-
IR Paragon 1000 (KBr). MALDI-TOF mass spectra were mea-
sured with a Bruker Biflex III with 19 kV acceleration voltage.
DHB (c = 10 µg/µL in 40% acetonitrile/water, 0.1% TFA) was used
Tris{2-[3Ј-[2ЈЈ,3ЈЈ,4ЈЈ-tri-O-acetyl-6ЈЈ-deoxy-6ЈЈ-(2ЈЈЈ-azipropylthio-
ureido)-α-D-mannopyranosyl]thioureido]ethyl}amine (5): The iso-
thiocyanato-functionalized glycocluster 3 (110 mg, 85 µmol) was
dissolved in dry dichloromethane (1 mL) and a solution of the di-
azirine 4 (31 mg, 0.25 mmol) in dry dichloromethane (500 µL) and
DIPEA (50 µL) was added dropwise. The reaction mixture was
stirred for 2 h at room temp., then the solvent was removed in
vacuo. Flash chromatography (ethyl acetate/cyclohexane, 3:1) of
the residue yielded the glycocluster 5 (70 mg, 44 µmol, 52%) as a
Eur. J. Org. Chem. 2006, 719–728
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M. Walter, M. Wiegand, T. K. Lindhorst
FULL PAPER
white powder. Rf = 0.79 (ethyl acetate). [α]D = +82.8 (c = 0.9 in
tracted with dichloromethane and the combined organic phases
1
CHCl3). H NMR (500 MHz, [D6]acetone): δ = 8.39 (br. s, 3 H, 3 were washed with water, dried with sodium sulfate and filtered.
NHman), 7.53, 7.39, 6.91, (each br. s, each 3 H, 9 NH), 6.08 (br. s,
3 H, 3 H-1), 5.40 (br. d, 3 H, 3 H-3), 5.31 (t~dd, 3 H, 3 H-2), 5.08
Column chromatography (cyclohexane/ethyl acetate, 3:1) gave 7
(170 mg, 0.99 mmol, 41 %) as a colorless syrup. 1H NMR
(t, 3 H, 3 H-4), 4.31–4.17 (br. m, 3 H, 3 H-6Ј), 4.11–4.02 (br. m, 3 (300 MHz, CDCl3): δ = 4.02 (dd, 2 H, Heq, HeqЈ), 3.71 (dd, 2 H,
H, 3 H-5), 3.93–3.80 [br. m, 3 H, 3 CHHC(NN)], 3.79–3.59 (br. m, Hax, HaxЈ), 3.50 (d, 2 H, CH2N3), 1.82 (dsept, 1 H, CH), 1.44 (s, 3
3
3
3 H, 3 H-6), 3.54–3.34 [m, 9 H, 3 NH–CH2CH2N, 3 CHHC(NN)], H, CH3), 1.40 (s, 3 H, CH3) ppm; JCHCHaxHeq = 5.2, JCHCHaxHeq
2.84–2.79 (br. m, 3 H, 3 NCHH), 2.65–2.58 (br. m, 3 H, 3 NCHH),
3
2
= 3.7, JCHCHHN = 7.3, JCHaxHeq
NMR (75.47 MHz, CDCl3): δ = 98.23 (Cq), 61.55 (2 CH2O), 50.81
=
2JCHaxЈHeqЈ = 12.1 Hz. 13C
2.16, 2.11, 2.08 [each s, each 9 H, 9 C(O)CH3], 1.07 (s, 9 H, 3
CH3) ppm; J2,3 = 3.7, J3,4 = 7.1, J4,5 = 7.1 Hz. 13C NMR (CH2N3), 34.18 (CH), 25.15, 22.42 (2 CH3) ppm. MS (CI): m/z =
3
3
3
(125.75 MHz, [D6]acetone): δ = 185.07, 184.01 (6 C=S), 170.31,
172.1 [M + H]+ (calcd. m/z = 172.2) for C7H14N3O3, m/z = 144.1
170.25, 170.16 (9 C=O), 78.73 (3 C-1), 73.78 (3 C-5), 69.37 (3 C- [M + H – N2]+ (calcd. m/z = 144.2) for C7H15NO2.
2), 68.96 (3 C-4), 68.64 (3 C-3), 52.59 (3 NCH2), 47.67 [3
CH2C(NN)], 45.14 (3 C-6), 43.36 (3 NCH2CH2), 26.72 [3 C(NN)],
2-Azidomethyl-1,3-propanediol (8): The dioxane 7 (204 mg,
1.2 mmol) was dissolved in MeOH (1 mL), mixed with concd. hy-
20.93, 20.76, 20.62 [9 C(O)CH3], 18.30 (3 CH3) ppm. ESI MS:
drochloric acid (200 µL) and stirred at room temp. for 1 h. After
m/z = 1566.4890 [M + H]+ (calcd. m/z = 1566.4721) for
complete deprotection the solution was neutralized with ion-ex-
C
57H88N19O21S6 m/z = 1588.5011; [M + Na]+ (calcd. m/z =
change resin (Merck III-OH–), filtered and the solution concen-
trated to provide 8 as a colorless syrup (156 mg, 1.2 mmol, quant.).
1H NMR (300 MHz, CD3OD): δ = 3.63 (t, 4 H, 2 CH2OH), 3.48
(d, 2 H, CH2N3), 1.90 (sept, 1 H, CH) ppm. 13C NMR (75.47 MHz,
CD3OD): δ = 61.26 (2 CH2OH), 50.91 (CH2N3), 45.03 (CH) ppm.
MS (CI): m/z = 132.1 [M + H]+ (calcd. m/z = 132.1) for
C4H10N3O2, 104.0 [M + H – N2]+ (calcd. m/z = 104.1) for
C4H10NO2.
1588.4540) for C57H87N19NaO21S6.
Tris{2-[3Ј[6ЈЈ-deoxy-6ЈЈ-(2ЈЈЈ-azipropylthioureido)-α-D-manno-
pyranosyl]thioureido]ethyl}amine (6): The acetylated glycocluster 5
(68 mg, 43 µmol) was dissolved in dry MeOH (500 µL), a solution
in sodium methoxide (1 m in MeOH, 39 µL, 0.1 equiv. per OH) was
added and the reaction mixture was stirred at 0 °C for 1 h. The
solution was neutralized by addition of ion-exchange resin (Am-
berlite IR 120), filtered, washed with MeOH and then the solvent
was removed in vacuo. The residue was purified by HPLC (Merck
RP-8, 7 µm; MeCN/H2O, 40:60). The purified fractions were lyo-
philized to yield the glycocluster 6 as a colorless amorphous solid
(36 mg, 30 µmol, 69%). Rf = 0.21 (MeOH/ethyl acetate, 1:1). [α]D
1,3-Bis(2Ј,3Ј,4Ј,6Ј-tetra-O-benzoyl-α-D-mannopyranosyloxy)-2-
(azidomethyl)propane (10): The azide 8 (87 mg, 0.66 mmol) and the
mannosyl donor 9 (1.09 g, 1.46 mmol) were dissolved in dry dichlo-
romethane (10 mL) under argon atmosphere at 0 °C. The glycosy-
lation reaction was started by the addition of TMS-OTf (10µL).
The solution was stirred at 0 °C for 1 h and then for another hour
at room temp. After quenching the reaction by addition of concd.
sodium hydrogen carbonate solution (1 mL) the mixture was di-
luted with dichloromethane/water (1:1, 50 mL). The organic phase
was separated and dried with sodium sulfate. After filtration and
concentration, column chromatography (cyclohexane/ethyl acetate,
3:1) gave 10 as a colorless solid (637 mg, 0.49 mmol, 75%). 1H
NMR (500 MHz, CDCl3): δ = 8.11, 8.03, 7.96, 7.81 (each dd, each
1
= –19.8 (c = 0.88 in MeOH). H NMR (500 MHz, [D6]acetone): δ
= 5.82, 5.64 (br. s, 3 H, 3 H-1), 4.06–3.50 [m, 30 H, 3 H-2, 3 H-3,
3 H-4, 3 H-5, 3 H-6, 3 H-6Ј, 3 NCH2CH2, 3 CH2C(NN)], 2.87–
2.74 (br. m, 6 H, 3 NCH2), 1.11, 1.10, 1.09 (s, 9 H, 3 CH3) ppm.
13C NMR (125.75 MHz, CD3OD): δ = 184.42 (6 C=S), 83.70 (3 C-
1), 77.93 (3 C-5), 75.07, 74.54 (3 C-3), 72.15, 72.03, 71.39 (3 C-2),
69.56, 68.98, 68.78 (3 C-4), 54.44, 54.09 (3 NCH2), 46.30 (3 C-6),
44.11 (3 NCH2CH2), 26.81 [3 C(NN)], 18.34, 18.28 (3 CH3) ppm.
ESI MS: m/z = 1188.3929 [M + H]+ (calcd. m/z = 1188.3770) for
C39H70N19O12S6; m/z = 1210.3760 [M + Na]+ (calcd. m/z =
1210.3590) for C39H69N19NaO12S6.
3
4 H, 16 HBz), 7.61–7.21 (m, 24 H, HBz), 6.14 (t, J4,5 = 10.1 Hz, 2
H, H-4man1, H-4man2), 5.93–5.88 (m, 3J3,4 = 10.1 Hz, 2 H, H-3man1
,
H-3man2), 5.77–5.74 (m, 3J2,3 = 3.3 Hz, 2 H, H-2man1, H-2man2), 5.17
(s, 3J1,2 = 1.8 Hz, 2 H, H-1man1, H-1man2), 4.77, 4.75 (each dd, 2J6,6Ј
= 12.2 Hz, each 1 H, H-6Јman1, H-6Јman2), 4.57, 4.55 (each dd, each
5-Azidomethyl-2,2-dimethyl-1,3-dioxane (7): 2-Hydroxymethyl-1,3-
propandiol (254 mg, 2.4 mmol) was dissolved in dry THF (5 mL)
and mixed with 2,2-dimethoxypropane (440 µL, 374 mg, 3.6 mmol)
and acetone (264 µL, 209 mg, 3.6 mmol). The reaction was started
by the addition of p-toluenesulfonic acid (50 mg) and the mixture
was stirred for 1.5 h at room temp. The reaction was quenched by
the addition of solid sodium carbonate (one spoon), finally the
liquid phase was filtered and concentrated. The residue was taken
up in dichloromethane/water, the aqueous phase was extracted
twice with dichloromethane, and the combined organic phases were
washed with water, dried with sodium sulfate and filtered. Without
further purification the concentrated residue was dissolved in dry
pyridine (10 mL), tosyl chloride (503 mg, 2.6 mmol) was added and
the reaction mixture was stirred overnight at room temp. Then
methanol (1 mL) was added and the solution concentrated in
vacuo. The residue was taken up in dichloromethane/water. The
aqueous phase was extracted with dichloromethane once, the com-
bined organic phases were washed with water, dried with sodium
sulfate and filtered. The concentrated residue was then dissolved in
dry toluene (10 mL), sodium azide (780 mg, 0.012 mol) and TBABr
3
3
1 H, H-6man1, H-6man2), 4.50–4.44 (m, J5,6 = 4.4, J5,6Ј = 3.9 Hz,
2 H, H-5man1, H-5man2), 4.02 (dd, 1 H, CHHOman1*), 3.97
(CHHOm a n 2 *), 3.74–3.64 (m, 4 H, CH2 N3 , CHHOm a n 1
,
CHHOman2), 2.44 [sept, 1 H, CH(CH2)3] ppm. 13C NMR
(125.75 MHz, CDCl3): δ = 166.15, 165.53, 165.38, 165.35 (8 C=O),
133.48, 133.42, 133.19, 133.11 [8 C(O)CBz], 129.89, 129.84, 129.74,
129.73, 129.40, 129.24, 129.02, 128.58, 128.48, 128.42, 128.30 (CBz),
98.07 (C-1man, C-1man2), 70.29 (C-2man1, C-2man2), 70.13, 70.10 (C-
3man1, C-3man2), 69.30 (C-5man1, C-5man2), 66.74 (C-4man1, C-4man2),
66.38, 66.30 (CH2Oman1, CH2man2), 62.83 (C-6man1, C-6man2), 49.88
(CH2N3), 39.17 [CH(CH2)3] ppm. MALDI-TOF MS (norharm-
ane): m/z = 1283.1 [M + Na – N2]+ (calcd. m/z = 1283.24) for
C72H61NNaO20; m/z = 1299.0 [M + K – N2]+ (calcd. m/z =
1299.19) for C72H61KNO20.
1,3-Bis(2,3,4,6-tetra-O-benzoyl-α-D-mannopyranosyloxy)-2-(isothio-
cyanatomethyl)propane (11): The azide 10 (300 mg, 0.23 mmol) was
dissolved in dry toluene (8 mL) and mixed with carbon disulfide
(773 mg, 2.4 mmol) were added and the reaction mixture was (560 µL, 707 mg, 9.3 mmol) and triethyl phosphite (160 µL,
stirred overnight at 65 °C. The mixture was concentrated, the resi-
due taken up in dichloromethane/water, the aqueous phase ex-
154 mg, 0.93 mmol). The solution was heated to 80 °C and stirred
under reflux for 8 h. After cooling to 0 °C water (5 mL) was added
724
www.eurjoc.org
© 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2006, 719–728

Synthesis of Cluster Mannosides Carrying a Photolabile Diazirine Group
and the reaction mixture was left standing overnight at 4 °C. solved in a 1:1 mixture of MeOH and THF (10 mL) and mixed
FULL PAPER
Dichloromethane (20 mL) was added, the organic phase was sepa-
rated, the aqueous phase was extracted with dichloromethane and
the combined organic phases were dried with sodium sulfate. After
filtration and concentration, flash column chromatography (cyclo-
hexane/ethyl acetate, 3:1) yielded 11 (170 mg, 0.13 mmol, 56%) as
with sodium methoxide solution (1 m in MeOH, 37 µL). The reac-
tion mixture was stirred for 1 h at room temp., neutralized with
Amberlite ion-exchange resin IR 120, filtered and purified by
HPLC (Merck RP-8, 7 µm; MeCN/H2O, 10:90). After lyophilis-
ation 13 was obtained as a colorless foam (17 mg, 30 µmol, 65%).
a yellowish solid. [α]2D0 = –16.1 (c = 0.88, CHCl ). IR (KBr): ν
1H NMR (500 MHz, D2O): δ = 4.83 (br. s, 2 H, 2 H-1), 4.02–3.43
˜
3
NCS
= 2104.4 cm–1. 1H NMR (500 MHz, CDCl3): δ = 8.07–7.14 (m, 40 (m, 20 H, 2 H-2, 2 H-3, 2 H-4, 2 H-5, 2 H-6, 2 H-6Ј, OCH2
,
man
3
H, HBz), 6.15–6.06 (m, J4,5 = 10.1 Hz, 2 H, H-4man1, H-4man2), CH2NH), 2.32 (br. s, 1 H, CH), 1.06 (s, 3 H, CH3) ppm. 13C NMR
3
3
5.81 (dd, J3,4 = 10.1 Hz, 2 H, H-3man1, H-3man2), 5.68 (dd, J2,3
=
(125.75, D2O): δ = 102.42 (C-1), 75.25 (C-5), 73.03 (C-3), 72.41 (C-
3.2 Hz, 2 H, H-2man1, H-2man2), 5.11 (d, J1,2 = 1.5 Hz, 1 H, H- 2), 69.18 (C-4), 68.77, 68.62 (OCH2man), 63.30 (C-6), 40.90 (CH),
3
1man1*), 5.09 (d, 1 H, H-1man2*), 4.74 –4.63 (m, J6Ј,6 = 12.3 Hz, 2
19.32 (CH3) ppm. MALDI-TOF MS (DHB): m/z = 551.4 [M +
3
H, H-6Јman1, H-6Јman2), 4.52–4.45 (m, 2 H, H-6man1, H-6man2), Na – N2]+ (calcd. m/z = 551.57) for C20H36N2NaO12S; m/z = 529.3
4.42–4.37 (m, J5,6Ј = 2.5 Hz, 2 H, H-5man1, H-5man2), 4.02–3.88 [M – N2]+ (calcd. m/z = 528.58) for C20H36N2O12S. ESI MS: m/z =
3
(m, 3 H, CH2Oman1*, CHH–NCS), 3.83–3.77 (m, 1 H, CHH– 579.1985 [M + Na]+ (calcd. m/z = 579.1943) for C20H36N4NaO12S.
NCS), 3.63 (dd, 1 H, CH2Oman2*), 2.50 (br. quint, 1 H, CH) ppm.
1,3-Bis(2Ј,3Ј,4Ј,6Ј-tetra-O-benzoyl-α-
azipropylthiourenyl-2ЈЈЈ,3ЈЈЈ,4ЈЈЈ-tri-O-acetyl-6ЈЈЈ-deoxy-6ЈЈЈ-thio-
urenylmethyl-α- -mannopyranos-6ЈЈЈ-yl)propane (16): The azide 10
D-mannopyranosyloxy)-2-(2ЈЈ-
13C NMR (125.75 MHz, CDCl3): δ = 165.54, 165.52, 165.51,
165.48, 165.34, 165.33 (8 C=O), 133.49, 133.44, 133.40, 133.21,
133.12, 133.09, 130.02, 129.94, 129.87, 129.85, 129.75, 129.71,
129.18, 129.17, 128.98, 128.95, 128.89, 128.62, 128.57,
D
(600 mg, 0.47 mmol) was dissolved in dry THF (5 mL) under argon
atmosphere at 0 °C and mixed with a solution of triphenylphos-
phane (244 mg, 0.93 mmol) in dry THF (2 mL). The mixture was
stirred at 0 °C for 0.5 h, then at room temp. for another 0.5 h. The
diazirine-labeled mannose derivative 15 (232 mg, 0.49 mmol), dis-
solved in dry THF (2 mL) and DIPEA (50 µL), was added to the
reaction mixture together with water (2 mL). The solution was
stirred at room temp. for 5 h, then concentrated in vacuo. Two puri-
fication steps (flash column chromatography first with cyclohex-
ane/ethyl acetate, 2:1 Ǟ 3:2, then cyclohexane/ethyl acetate, 1:1)
led to the pure product (188 mg, 0.11 mmol, 23%) as an amorph-
ous solid. [α]2D0 = –4.1 (c = 1.25, H2O). 1H NMR (500 MHz,
CDCl3): δ = 8.18–7.27 (m, 40 H, HBz), 8.11 (br. s, 1 H, NH), 7.20
(br. s, 1 H, NH), 6.29–6.22 (m, 2 H, H-4man1, H-4Bzman2), 6.11 (br.
s, 1 H, H-1man3), 6.03–5.97 (m, 2 H, H-3man1, H-3man2), 5.91–5.87
128.49,128.42, 128.41, 128.31, 128.29 (CBz), 98.18, 98.09 (C-1man1
,
C-1man2), 70.18, 70.15 (C-3man1, C-3man2), 70.11, 70.05 (C-2man1, C-
2man2), 69.41, 69.39 (C-5man1, C-5man2), 66.59, 66.52 (C-4man1, C-
4man2), 65.90, 65.85 (CH2Oman1, CH2Oman2), 62.70, 62.66 (C-6man1
,
C-6man2), 43.76 (CH2NCS), 39.72 (CH) ppm. MALDI-TOF MS:
m / z = 1 3 2 7 . 0 [ M + N a ] + ( c a l c d . m / z = 1 3 2 6 . 3 4 ) f o r
C73H61NNaO20S; m/z = 1342.9 [M + K]+ (calcd. m/z = 1342.29 for
C73H61KNO20S).
2-(2Ј-Azipropylthiourenyl)-1,3-bis(2ЈЈ,3ЈЈ,4ЈЈ,6ЈЈ-tetra-O-benzoyl-α-
D-mannopyranosyloxy)propane (12): The isothiocyanate 11 (149 mg,
0.11 mmol) was dissolved in dry dichloromethane (2 mL) and
mixed with 4 (14 mg, 0.11 mmol) and a solution (50 µL) of DIPEA
in dry dichloromethane (0.5 mL). The reaction mixture was strirred
at room temp. for 4 h, then the solvent was removed in vacuo. Puri-
fication of the residual syrup by column chromatography (cyclo-
hexane/ethyl acetate, 2:1) yielded 12 (76 mg, 0.06 mmol, 48%) as a
colorless solid. [α]2D0 = –14.6 (c = 0.52, CHCl3). 1H NMR
(500 MHz, CDCl3): δ = 8.13–7.15 (m, 40 H, HBz), 6.81 (NHdiazirine),
(m, 2 H, H-2man1, H-2man2), 5.38–5.30 (m, 3 H, H-1man1, H-1man2
,
H-3man3), 5.27 (tϷdd, 1 H, H-2man3), 5.07 (t, 1 H, H-4man3), 4.87–
5.75 (m, 4 H, H-6Јman1, H-6Јman2, H-5man1, H-5man2), 4.70–4.62 (m,
2 H, H-6man1, H-6man2), 4.27–4.19 (m, 2 H, CHHOm an 1
,
CHHOman2), 4.06–3.90 (m, 6 H, H-5man3, H-6man3, H-6Јman3, CH–
6.62 (NH–CH2core), 6.22 (t, 3J4,5 = 10.2 Hz, 2 H, H-4man1, H-4man2), CHHNH, CHHOm a n 1 , CHHOm a n 2 ), 3.77–3.63 [m, 3 H,
3
3
5.88 (dd, J3,4 = 10.2 Hz, 1 H, H-3man1*), 5.82–5.75 (m, J2,3
=
CH2C(NN), CH–CHHNH], 2.73 (sept, 1 H, CH), 2.04, 1.98, 1.95
3.1 Hz, 3 H, H-2man1, H-2man2, H-3man2*), 5.20 (d, J1,2 = 1.5 Hz, [each br. s, each 3 H, 3 C(O)CH3], 1.04 (s, 3 H, CH3) ppm; man1,2:
3
1 H, H-1man1*), 5.18 (d, 1 H, H-1man2*), 4.83–4.73 (m, J6,6Ј
=
3J1,2 = 1.6, 3J2,3 = 3.3, 3J3,4 = 9.9, 3J4,5 = 9.9; man3: 3J2,3 = 3.4, 3J3,4
2
12.1 Hz, 2 H, H-6Јman1, H-6Јman2), 4.63–4.54 (m, J5,6 = 3.9, J5,6Ј = 7.9, 3J4,5 = 7.9; core: 3JCHCHH = 5.7 Hz. 13C NMR (125.75 MHz,
= 2.4 Hz, 4 H, H-5man1, H-5man2, H-6man1, H-6man2), 4.36–4.26 [m, CDCl3): δ = 170.22, 170.13, 170.09 (3 C=OAc), 166.43, 166.39,
1 H, NHCHHC(NN)], 4.21 (dd, 1 H, CHHOman1*), 4.12 (dd, 1 H, 166.01, 165.86, 165.85, 165.72, 165.71 (8 C=OBz), 134.40, 134.19,
CHHOman2*), 3.97–3.89 [m, 1 H, NHCHHC(NN)], 3.83–3.74 (m, 134.15, 134.09, 134.04, 134.02, 131.05, 130.43, 130.42, 130.40,
3
3
3 H, CHHOman1, CHHOman2, CHHNH), 3.59 (dd, 1 H, CHHNH),
2.56 (sept, 1 H, CH), 1.06 (s, 3 H, CH3) ppm. 13 C NMR
(125.75 MHz, CDCl3): δ = 184.10 (C=S), 166.27, 166.20, 166.16,
130.37, 130.36, 130.34, 130.19, 130.18, 130.15, 129.57, 129.51,
129.30, 129.29, 129.21, 129.19 (CBz), 99.09, 98.97 (C-1man1, C-
1man1), 79.97 (C-1man3), 71.66 (C-3man1, C-3man2), 71.23, 71.15 (C-
165.54, 165.51, 165.46 (8 C=O), 133.55, 133.49, 133.38, 133.36, 2man1, C-2man2), 69.97, 69.92 (C-5man1, C-5man2), 69.74 (C-2man3),
133.26, 133.13, 133.07, 129.90, 129.88, 129.77, 129.76, 129.71, 69.40 (C-3man3), 68.54 (C-4man3), 67.95, 67.88 (CH2Oman1
,
129.14, 129.04, 128.95, 128.87, 128.58, 128.57, 128.51, 128.48, CH2Oman2), 67.29 (C-4man1, C-4man1), 63.35 (C-6man1, C-6man2),
128.46, 128.37, 128.30, 128.29 (CBz), 98.09, 98.01 (C-1man1, C- 47.76 [CH2C(NN)], 45.29 (C-6man3), 43.44 (CHCH2NH), 40.10
1man2), 70.74, 70.68 (C-3man1, C-3man2), 70.34, 70.22 (C-2man1, C-
2man2), 69.87 (CH2Oman1*), 69.61, 69.46 (C-5man1, C-5man2), 69.28
(CH2Oman2*), 66.30, 66.26 (C-4man1, C-4man2), 62.76, 62.73 (C-
6man1, C-6man2), 47.72 (CH2NHcore), 46.43 [NHCH2C(NN)], 38.81
(CH), 25.85 [C(NN)], 18.19 (CH3) ppm. MALDI-TOF MS (nor-
harmane): m/z = 1384.7 [M + Na – N2]+ (calcd. m/z = 1384.41) for
C76H68N2NaO20S, m/z = 1361.7 [M – N2]+ (calcd. m/z = 1361.47)
for C76H68N2O20S.
(CH), 26.45 [C(NN)], 20.72, 20.64, 20.53 [3 C(O)CH3], 18.30
(CH3) ppm.
1,3-Bis(α-
oxy-6ЈЈ-thiourenylmethyl-α-
D
-mannopyranosyloxy)-2-(2Ј-azipropylthiourenyl-6ЈЈ-de-
-mannopyranos-6ЈЈ-yl)propane (17):
D
The protected cluster 16 (180 mg, 0.1 mmol) was dissolved in dry
MeOH (2 mL) and sodium methoxide (1 m in MeOH, 114 µL) was
added. The reaction mixture was stirred at room temp. or 1 h, then
the solution was neutralized with Amberlite ion exchanger IR 120,
[2-(2Ј-Azipropylthiourenyl)-1,3-bis(α-
propane (13): The protected derivative 12 (65 mg, 47µmol) was dis-
D
-mannopyranosyloxy)]- filtered, and concentrated. The residue was purified by HPLC
(Merck RP-8, 7 µm; MeCN/H2O, 15:85). Lyophilisation led to 17
Eur. J. Org. Chem. 2006, 719–728
© 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
725

M. Walter, M. Wiegand, T. K. Lindhorst
FULL PAPER
(51 mg, 0.07 mmol, 63%) as a colorless foam. [α]2D0 = +39.8 (c =
0.88, H2O). H NMR (500 MHz, D2O): δ = 5.50 (br. s, 1 H, NH),
Tri-O-(2-acetoxyethyl)-O-tosylpentaerythritol (21): To a solution of
the triol 20 (2.20 g, 5.0 mmol) in dry pyridine (20 mL) acetic anhy-
dride (2.8 mL, 0.03 mol) was added and the reaction mixture was
stirred at room temperature for 4 h. Then it was concentrated,
washed with concd. aqueous sodium hydrogen carbonate solution,
dried with sodium sulfate and filtered. Purification by column
chromatography (cyclohexane/ethyl acetate, 3:2) yielded 21 (2.40 g,
4.0 mmol, 84%) as a colorless syrup. 1H NMR (300 MHz, CDCl3):
δ = 7.78 (d, 3Ja,x = 3JaЈ,xЈ = 8.3 Hz, 2 H, Ha,aЈ), 7.35 (d, 2 H, Hx,xЈ),
4.14–4.08 (m, 6 H,3 CH2OAc), 4.03 (s, 2 H, CH2OTs), 3.56–3.50
1
4.86–4.81 (m, 2 H, H-1man3, H-1man3*), 4.02–3.93 (m, 3 H, H-2man1
,
H-2man2, H-2man3), 3.87 (dϷdd, 2 H, H-6Јman1, H-6Јman2), 3.84–3.47
[m, 21 H, H-3man1, H-3man2, H-3man3, H-4man1, H-4man2, H-4man3
,
H-5man1, H-5man2, H-5man3, H-6man1, H-6man2, H-6man3, H-6Јman3
,
CH2Oman1, CH2Oman2, CHCH2NH, CH2C(NN)], 2.33 (br. s, 1 H,
3
3
CH), 1.07 (br. s, 3 H, CH3) ppm; man1,2: J1,2 = 1.6, J2,3 = 3.3,
3J3,4 = 9.7, J4,5 = 9.7, J5,6 = 5.7, J6,6Ј = 11.9 Hz. 13C NMR
3
3
2
(125.75 MHz, D2O): δ = 102.46, 102.44 (C-1man1, C-1man2), 84.44
(C-1man3), 75.33 (C-5man3), 75.25 (C-5man1, C-5man2), 73.04 (C- (m, 6 H, 3 CH2CH2OAc), 3.40 [s, 6 H, C(CH2O)3], 2.46 (s, 3 H,
3man1, C-3man2), 72.67 (C-3man3), 73.43 (C-2man1, C-2man2), 70.05
(C-4man3), 69.17 (C-4man1, C-4man2), 68.78, 68.68 (CH2Oman1
CH3), 2.06 [s, 9 H, 3 C(O)CH3] ppm. 13C NMR (75.47 MHz,
CDCl3): δ = 170.96 (3 C=O), 144.79 (H3CCaryl), 132.86 (O3SCaryl),
,
CH2Oman2), 63.29 (C-6man1, C-6man2), 48.57 [CH2C(NN)], 46.40 (C- 129.79 [H3CCaryl(Cx,xЈ)], 127.94 [O3SCaryl(Ca,aЈ)], 69.22 (3 OCH2-
6man3), 40.98 (CH), 28.86 [C(NN)], 19.46 (CH3) ppm. ESI MS:
m/z = 799.2506 [M + Na]+ (calcd. m/z = 799.2460) for
C27H48N6NaO16S2.
CH2OAc), 69.16 (TsOCH2), 68.72 [C(CH2O)3], 61.37 (3 OCH2-
CH2OAc), 44.95 (Cq), 21.65 (H3CCaryl), 20.90 [3 C(O)CH3] ppm.
MALDI-TOF MS: m/z = 571.5 [M + Na]+ (calcd. m/z = 571.18),
587.5 [M + K]+ (calcd. m/z = 587.15) for C24H36O12S.
Tri-O-allyl-O-tosylpentaerythritol (19): Triallyl pentaerythritol (18,
4.00 g, 16 mmol) was dissolved in dry pyridine (30 mL). Tosyl chlo-
ride (3.24 g, 17 mmol) was added in portions to this solution and
the mixture stirred at room temp. overnight. The reaction was
quenched by addition of methanol (10 mL), the solution concen-
trated and the residue taken up in dichloromethane/concd. aqueous
sodium hydrogen carbonate solution. The organic phase was sepa-
rated, washed once with concd. aqueous sodium hydrogen carbon-
ate solution and dried with sodium sulfate. After filtration and col-
umn chromatography (cyclohexane/ethyl acetate, 9:1) 19 was ob-
tained (5.30 g, 13 mmol, 81 %) as a yellowish syrup. 1H NMR
(300 MHz, CDCl3): δ = 7.77 (d, 2 H, Ha,aЈ), 7.33 (d, 2 H, Hx,xЈ),
1-Azido-3-(2Ј-acetoxyethoxy)-2,2-bis(2Ј-acetoxyethoxymethyl)pro-
pane (22): To a solution of the tosylate 21 (2.20 g, 4.0 mmol) in dry
DMF (15 mL) sodium azide (1.30 g, 0.02 mol) and TBABr (1.29 g,
4.0 mmol) were added. The reaction mixture was stirred for 6 d at
85 °C. The solution was concentrated and the residue taken up in
water/ethyl acetate. The organic phase was separated, washed twice
with water, dried with sodium sulfate and filtered. After concentra-
tion in vacuo and column chromatography (cyclohexane/ethyl ace-
tate, 2:1) 22 was obtained (1.57 g, 3.7 mmol, 93%) as a colorless
syrup. 1H NMR (300 MHz, CDCl3): δ = 4.20 (dt, 6 H, 3 CH2OAc),
3.61 (dt, 6 H, 3 OCH2CH2OAc), 3.40 [s, 6 H, 3 C(CH2O)3], 3.35
(s, 2 H, CH2N3), 2.07 [s, 9 H, 3 C(O)CH3] ppm. 13C NMR
(75.47 MHz, CDCl3): δ = 171.00 (3 C=O), 69.53 [C(CH2O)3], 69.27
(3 OCH2CH2OAc), 63.34 (3 CH2OAc), 51.50 (CH2N3), 45.49 (Cq),
20.93 [3 C(O)CH3] ppm. MS (CI): m/z = 392.2 [M + H – N2]+
(calcd. m/z = 392.19) for C17H30NO9.
5.79 (dddd, 3 H, 3 OCH2CH=CH2), 5.18 (dddϷdd, 2Jgem = 1.8 Hz,
3
3Jtrans = 17.3 Hz, 3 H, 3 OCH2CH=CHHt), 5.12 (dddϷdd, Jcis
=
10.5 Hz, 3 H, 3 OCH2CH=CHcH), 4.06 (s, 2 H, TsOCH2), 3.86
3
(dt, JOCHHCH = 5.4 Hz, 6 H, 3 OCH2CH=CH2), 3.38 [s, 6 H, 3
C(CH2O)3], 2.44 (CH3) ppm. 13C NMR (125.75 MHz, CDCl3): δ =
144.53 (H3CCaryl), 134.76 (3 CH=CH2), 132.90 (O3SCaryl), 129.73
[H3CCaryl(Cx,xЈ)], 127.96 [O3SCaryl(Ca,aЈ)], 116.39 (3 CH=CH2),
72.28 (3 OCH2CH=CH2), 69.74 (TsOCH2), 68.18 [3 C(CH2O)3],
44.81 (Cq), 21.61 (CH3) ppm.
1-Azido-3-(2Ј-hydroxyethoxy)-2,2-bis(2Ј-hydroxyethoxymethyl)pro-
pane (23): The azide 22 (1.50 g, 3.6 mmol) was dissolved in dry
MeOH (5 mL), mixed with sodium methoxide (1 m in MeOH,
1 mL) and stirred at room temp. for 30 min. Then the solution was
neutralized with Amberlite ion-exchange resin IR 120. After fil-
tration, removal of the solvent and purification by column
chromatography (ethyl acetate/cyclohexane, 2:1 Ǟ MeOH/ethyl
acetate, 2:1) 23 was obtained (952 mg, 3.3 mmol, 91%) as a yellow-
Tri-O-(2-hydroxyethyl)-O-tosylpentaerythritol (20): The tosylate 19
(5.00 g, 12 mmol) was dissolved in a 1:1 mixture of dry dichloro-
methane and dry methanol (40 mL), the solution was cooled to
–78 °C. Oxygen was passed through the solution for 5 min, then
ozone for 40 min, then again oxygen for 5 min and finally argon
for 5 min. Then sodium boronhydride (4.54 g, 0.12 mol) was added
in portions to the cold solution. The reaction mixture was stirred
for 2 h at 0 °C and was then left standing at room temp. overnight.
The reaction was quenched by addition of acetic acid (15 mL), the
solution concentrated and the residue taken up in diethyl ether/
1
ish syrup. H NMR (300 MHz, CDCl3): δ = 3.76–3.69 (m, 6 H, 3
CH2OH), 3.61–3.54 (m, 6 H, 3 OCH2CH2OH), 3.46 [s, 6 H,
C(CH2O)3], 3.39 (s, 2 H, CH2N3), 2.98 (br. s, 3 H, 3 OH) ppm. 13
C
NMR (75.47 MHz, CDCl3): δ = 72.63 (3 OCH2CH2OH), 70.02
[C(CH2O)3], 61.41 (3 CH2OH), 51.91 (CH2N3), 45.34 (Cq) ppm.
1-Azido-3-[2Ј-(2ЈЈ,3ЈЈ,4ЈЈ,6ЈЈ-tetra-O-benzoyl-α-
D
-mannopyranosyl-
-manno-
pyranosyloxy)ethoxymethyl]propane (24): The triol 23 (100 mg,
water. The aqueous phase was extracted twice with diethyl ether oxy)ethoxy]-2,2-bis[2Ј-(2ЈЈ,3ЈЈ,4ЈЈ,6ЈЈ-tetra-O-benzoyl-α-
D
and the combined organic phases were washed once with water.
After drying with sodium sulfate and filtration, column chromatog-
raphy (ethyl acetate/cyclohexane, 2:1 Ǟ ethyl acetate Ǟ ethyl ace-
tate/methanol, 2:1) the triol 20 (2.32 g, 5.0 mmol, 46%) was ob-
0.34 mmol) and 9 (833 mg, 1.12 mmol) were dissolved in dry
dichloromethane (10 mL) under argon and cooled to 0 °C. The re-
action was started by addition of TMS-OTf (10µL). The mixture
was stirred at 0 °C for 1 h, then for an additional day at room temp.
1
tained as a colorless syrup. H NMR (300 MHz, CDCl3): δ = 7.78
3
3
(d, Ja,x = JaЈ,xЈ = 8.3 Hz, 2 H, Ha,aЈ), 7.36 (d, 2 H, Hx,xЈ), 4.07 (s, The reaction was quenched by addition of a small amount of
2 H, CH2OTs), 3.69–3.63 (m, 6 H, 3 CH2OH), 3.53–3.47 (m, 6 H, concd. aqueous sodium hydrogen carbonate solution. The mixture
3 CH2CH2OH), 3.45 [s, 6 H, 3 C(CH2O)3], 3.02 (br. s, 3 H, 3 OH),
was taken up in dichloromethane/water, the organic phase sepa-
rated, dried with sodium sulfate and filtered. After removal of the
2.46 (s, 3 H, CH3) ppm. 13C NMR (125.75 MHz, CDCl3): δ =
145.01 (H3CCaryl), 132.71 (O3SCaryl), 129.91 [H3CCaryl(Cx,xЈ)], solvent and purification by column chromatography (cyclohexane/
127.91 [O3SCaryl(Ca,aЈ)], 72.67 (3 OCH2CH2OH), 69.10 (TsOCH2), ethyl acetate, 5:2) 24 was obtained (466 mg, 0.23 mmol, 67%) as a
69.04 [C(CH2O)3], 61.37 (3 OCH2CH2OH), 44.95 (Cq), 21.65 colorless solid. [α]2D0 = –20.7 (c = 1.98, CHCl3). 1H NMR
(CH3) ppm.
(500 MHz, CDCl3): δ = 8.10, 8.03, 7.93, 7.82 (each dd, each 4 H,
726
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© 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2006, 719–728

Synthesis of Cluster Mannosides Carrying a Photolabile Diazirine Group
FULL PAPER
16 HBz), 7.58–7.53 (m, 4 H, 4 HBz), 7.47–7.29 (m, 24 H, HBz), 7.25–
7.21 (m, 6 H, 6 HBz), 6.13 (t, 3 H, 3 H-4), 5.96 (dd, 3 H, 3 H-3),
H, H-4), 5.94 (dd, J3,4 = 10.1 Hz, 3 H, H-3), 5.73 (dd, J2,3
=
3
3
3.2 Hz, 3 H, H-2), 5.20 (d, 3J1,2 = 1.6 Hz, 3 H, H-1), 4.73 (dd, 3J5,6Ј
3
2
5.73 (dd, 3 H, 3 H-2), 5.17 (d, 3 H, 3 H-1), 4.72 (dd, 3 H, 3 H-6Ј), = 2.5 Hz, 3 H, H-6Ј), 4.54–4.47 (m, J5,6 = 4.1, J6,6Ј = 12.1 Hz, 6
4.54–4.47 (m, 6 H, 3 H-5, 3 H-6), 3.94 (ddd, 3 H, 3 man-
H, H-5, H-6), 3.99–3.92 (m, 3 H, manOCHHCH2), 3.85–3.61 [m,
19 H, manOCHHCH2, manOCH2CH2, CH2NHC(S), C(CH2N)],
OCHHCH2), 3.79 (ddd, 3 H, 3 manOCHHCH2), 3.73 (q, 6 H, 3
manOCH2CH2), 3.57 [d, 6 H, 3 C(CH2O)3], 3.47 (d, 2 H, 3.61 [s, 6 H, C(CH2O)3], 0.99 [s, 3 H, C(NN)CH3] ppm. 13C NMR
3
3
3
CH2N3) ppm; Ja,x
=
3JaЈ,xЈ = 8.4, J1,2 = 1.8, J2,3 = 3.3, (125.75 MHz, CDCl3): δ = 166.17, 165.53, 165.49, 165.42 (C=O),
3J3,4 = 10.1, J4,5 = 10.1, J5,6 = 4.2, J5,6Ј = 2.2, J6,6Ј = 11.5, 133.42, 133.40, 133.15, 133.06 [OC(O)CBz], 129.89, 129.85, 129.78,
3
3
3
2
2JCHHCHHOman = 11.2, JCHHCHHOman = 5.6, JCHHCHHOman
=
129.74, 129.29, 129.08, 128.99, 128.56, 128.47, 128.30 (CBz), 97.67
(C-1), 70.62 (manOCH2CH2), 70.60 C(CH2O)3) , 70.52 (C-2), 70.16
(C-3), 68.90 (C-5), 67.28 (manOCH2CH2), 66.89 (C-4), 62.87 (C-
6), 43.47 (Cq) ppm. MALDI-TOF MS (norharmane): m/z = 2102.1
[M – N2]+ (calcd. m/z = 2102.17) for C117H108N2O33S.
3
3
8.8 Hz. 13C NMR (125.75 MHz, CDCl3): δ = 165.14, 165.49,
165.38, 165.30 (12 C=O), 133.40, 133.37, 133.07, 133.03 [OC(O)-
CBz], 129.94, 129.84, 129.75, 129.73, 129.40, 129.17, 129.04, 128.55,
128.45, 128.43, 128.27 (CBz), 97.66 (3 C-1), 70.50 (3 man-
OCH2CH2), 70.48 (3 C-2), 70.11 (3 C-3), 69.91 [C(CH2O)3], 68.79
(3 C-5), 67.17 (3 manOCH2CH2), 67.00 (3 C-4), 62.87 (3 C-6),
51.90 (CH2N3), 45.73 (Cq) ppm. MALDI-TOF MS (norharmane):
m / z = 2 0 5 2 . 3 [ M – N a ] + ( c a l c d . m / z = 2 0 5 2 . 0 ) f o r
C117H108N2NaO33S; m/z = 2068.2 [M + K]+ (calcd. 2068.0) for
C117H108KN2O33S.
N-{3-[2Ј-(α-D-Mannopyranosyloxy)ethoxy]-2,2-bis[2Ј-(α-D-manno-
pyranosyloxy)ethoxymethyl]propyl}-NЈ-(2ЈЈ-azipropyl)thiourea (27):
The protected cluster 26 (225 mg, 0.11 mmol) was dissolved in a
1:1 mixture of dry MeOH and THF (2 mL) and mixed with sodium
methoxide (1 m in MeOH, 126 µL). The solution was stirred for 1 h
at room temp., then neutralised with Amberlite ion-exchange resin
IR 120. After filtration and removal of the solvent the residue was
purified by column chromatography (RP-18, water/MeOH, 5:1,
then lyophilisation) to yield 27 (51 mg, 0.06 mmol, 55%) as a color-
less foam. [α]2D0 = 21.3 (c = 0.70, H2O). 1H NMR (500 MHz, D2O):
δ = 4.90 (d, 3 H, 3 H-1), 3.98–3.94 (m, 3 H, 3 H-2), 3.90–3.61
[m, 31 H, 3 H-3, 3 H-4, 3 H-5, 3 H-6, 3 H-6Ј, 3 manO–CH2CH2,
CH2NHC(S)], 3.56–3.46 [m, 6 H, C(CH2O)3], 3.34 [s, 2 H,
C(CH2N)], 1.08 [s, 3 H, C(NN)CH3] ppm. 13C NMR (125.75 MHz,
D2O): δ = 102.23 (3 C-1), 75.14 (3 C-4), 73.02 (3 C-3), 72.74 (3
manO–CH2CH2), 72.44 (3 C-2), 69.12 (3 C-5), 68.63 [C(CH2O)3],
64.87 (3 manO–CH2), 51.26 [CH2NHC(S)], 46.79 (Cq), 26.25
[C(NN)], 19.37 [C(NN)CH3] ppm. MALDI-TOF MS (DHB):
m/z = 875.5 [M + Na – N2 ]+ (calcd. m/z = 875.33) for
C33H60N2NaO21S. ESI MS: m/z = 903.3358 [M + Na]+ (calcd.
m/z = 903.3363) for C33H60N4NaO21S.
1-Isothiocyanato-3-[2Ј-(2ЈЈ,3ЈЈ,4ЈЈ,6ЈЈ-tetra-O-benzoyl-α-
D-manno-
pyranosyloxy)ethoxy]-2,2-bis[2Ј-(2ЈЈ,3ЈЈ,4ЈЈ,6ЈЈ-tetra-O-benzoyl-α-
D-
mannopyranosyloxy)ethoxymethyl]propan (25): The azide 24
(400 mg, 0.20 mmol) was dissolved in dry toluene (5 mL) and
mixed with carbon disulfide (480 µL, 600 mg, 8 mmol) and triethyl
phosphite (136 µL, 131 mg, 0.80 mmol). The reaction mixture was
heated to 80 °C and stirred at reflux temperature for 8 h. After
cooling, water (2 mL) was added and the reaction mixture was left
standing overnight at 4 °C. The organic phase was diluted with
dichloromethane and the aqueous phase extracted once with
dichloromethane. The combined organic phases were dried with
sodium sulfate. The solution was filtered, concentrated and the resi-
due purified by column chromatography (cyclohexane/ethyl acetate,
3:2) to yield 25 (333 mg, 0.16 mmol, 83 %) as a colorless solid.
1
[α]2D0 = –41.4 (c = 0.64, CHCl3). H NMR (500 MHz, CDCl3): δ =
8.09, 8.03, 7.93, 7.82 (each dd, each 4 H, HBz), 7.58–7.52 (m, 4 H,
4 HBz), 7.47–7.29 (m, 24 H, HBz), 7.25–7.20 (t, 6 H, 6 HBz), 6.13
1,9-Bis(2Ј,3Ј:4Ј,6Ј-diisopropylidene-α-D-mannopyranosyloxy)-3,7-di-
3
3
oxanonan-5-one (29): The alkene 28 (1.44 g 2.2 mmol) was dissolved
in a 1:1 mixture of dry dichloromethane and methanol (20 mL)
and cooled to –78 °C. Oxygen was passed through the reaction mix-
ture for 5 min, then ozone for 20 min, then again oxygen for 5 min
and finally argon for 5 min. Then triphenylphosphane (2.22 g,
8.5 mmol) was added to the cold solution which was then stirred
for 2 h at room temp. After evaporation of the solvent in vacuo the
residue was purified by flash chromatography (cyclohexane/ethyl
acetate, 2:1) to yield 29 (1.33 g, 2.0 mmol, 92%) as an amorphous
colorless solid. [α]2D0 = +6.8 (c = 1.25, CHCl3). 1H NMR (500 MHz,
CDCl3): δ = 5.07 (s, 2 H, 2 H-1), 4.31 (d, 8 H, 4 Hc,cЈ), 4.22 (d,
(t, J4,5 = 10.0 Hz, 3 H, 3 H-4), 5.95 (dd, J3,4 = 10.0 Hz, 3 H, 3
3
3
H-3), 5.73 (dd, J2,3 = 3.3 Hz, 3 H, 3 H-2), 5.17 (d, J1,2 = 1.9 Hz,
3 H, 3 H-1), 4.72 (dd, J6,6Ј = 13.0 Hz, 3 H, 3 H-6), 4.53–4.46 (m,
2
3J5,6 = 3.5 Hz, 6 H, 3 H-5, 3 H-6Ј), 3.97–3.91 (m, 3 H, 3 manO–
CHH), 3.83–3.68 (m, 11 H, 3 manOCHH, 3 manOCH2CH2,
CH2NCS), 3.60 [d, 6 H, C(CH2O)3] ppm. 13C NMR (125.75 MHz,
CDCl3): δ = 166.12, 165.48, 165.38, 165.29 (12 C=O), 133.39,
133.36, 133.06, 133.02 [OC(O)CBz], 129.90, 129.83, 129.74, 129.73,
129.36, 129.14, 129.00, 128.53, 128.44, 128.42, 128.25 (CBz), 97.66
(3 C-1), 70.60 (3 manO–CH2CH2), 70.47 (3 C-2), 70.08 (3 C-3),
69.70 [C(CH2O)3], 68.81 (3 C-5), 67.12 (3 manO–CH2), 66.98 (3 C-
4), 62.87 (3 C-6), Cq (46.03), 45.78 (CH2NCS) ppm. MALDI-TOF
MS (norharmane): m/z = 2068.0 [M – Na]+ (calcd. m/z = 2068.06)
for C117H108N2NaO33S.
3
3J2,3 = 5.7 Hz, 2 H, 2 H-2), 4.16 (dd, J3,4 = 7.9 Hz, 2 H, 2 H-3),
3.89–3.82 (m, 4 H, 2 H-6, 2 Ha), 3.79–3.64 (m, 10 H, 2 H-4, 2 H-
2
3
6Ј, J6,6Ј = 10.8 Hz, 4 Hb,bЈ, 2 HaЈ), 3.59 (dt, J5,6 = 5.6 Hz, 2 H, 2
H-5), 1.55, 1.52, 1.42, 1.35 (each s, each 6 H, 8 iPr-CH3) ppm. 13C
NMR (125.75, CDCl3): δ = 205.59 (C=O), 109.46 (2 2,3-iPr-Cq),
99.70 (2 4,6-iPr-Cq), 97.92 (2 C-1), 75.94 (2 C-2), 75.01 (2 Cc), 74.80
(2 C-3), 72.68 (2 C-4), 70.73 (2 Cb), 66.56 (2 Ca), 62.02 (2 C-6),
61.45 (2 C-5), 29.04, 28.15, 26.14, 18.77 (8 iPr-CH3) ppm. MALDI-
TOF MS (norharmane): m/z = 685.75 [M + Na]+ (calcd. m/z =
685.30) for C31H50NaO15, m/z = 701.71 [M – K]+ (calcd. m/z =
701.25) for C31H50KO15.
N-{3-[2Ј-(2ЈЈ,3ЈЈ,4ЈЈ,6ЈЈ-Tetra-O-benzoyl-α-
ethoxy]-2,2-bis[2Ј-(2ЈЈ,3ЈЈ,4ЈЈ,6ЈЈ-tetra-O-benzoyl-α-
D
-mannopyranosyloxy)-
-mannopyranos-
D
yloxy)ethoxymethyl]propyl}-NЈ-(2ЈЈЈ-azipropyl)thiourea (26): The
isothiocyanate 25 (318 mg, 0.16 mmol) was dissolved in dry dichlo-
romethane (3 mL). DIPEA (50 µL) and a solution of the diazirine
4 (13 mg, 0.16 mmol) in dichloromethane (0.5 mL) were added and
the reaction mixture was stirred at 30 °C for 8 h. The solvent was
removed in vacuo and purification of the residue by column
chromatography (cyclohexane/ethyl acetate, 2:1) yielded 26
(265 mg, 0.12 mmol, 80%) as a colorless solid. [α]2D0 = –35.0 (c =
1,9-Bis(2Ј,3Ј:4Ј,6Ј-diisopropylidene-α-D-mannopyranosyloxy)-5-azi-
3,7-dioxanonane (30): The ketone 29 (810 mg, 1.22 mmol) was dis-
solved in a 1:2 mixture of dry dichloromethane and methanol
1
0.16, CHCl3). H NMR (500 MHz, CDCl3): δ = 8.09, 8.02, 7.93,
7.82 (each dd, each 6 H, HBz), 7.58–7.53 (m, 6 H, HBz), 7.47–7.28 (5 mL). After cooling of the solution to –50 °C gaseous NH3 was
3
(m, 24 H, HBz), 7.25–7.21 (t, 6 H, HBz), 6.14 (t, J4,5 = 10.1 Hz, 3
added until a volume of approx. 80 mL had condensed. The solu-
Eur. J. Org. Chem. 2006, 719–728
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727

M. Walter, M. Wiegand, T. K. Lindhorst
FULL PAPER
tion was stirred for 4 h at –20 °C. Then hydroxylamine-O-sulfonic
acid (152 mg, 1.24 mmol), dissolved in dry methanol (2 mL), was
added slowly at –50 °C. Cooling was stopped after 1 h and the solu-
tion was left standing overnight at room temp. The solution was
filtered and concentrated. The remaining residue was dissolved in
methanol (20 mL) and triethylamine (10 mL). This solution was
titrated with a 10% methanolic iodine solution until the colour
remained yellow. This solution was again concentrated and the resi-
due dissolved in diethyl ether and water. The aqueous phase was
extracted twice with diethyl ether and the combined organic phases
were washed once with aqueous sodium thiosulfate solution and
once with water. After drying with sodium sulfate and filtration
purification by column chromatography (cyclohexane/ethyl acetate,
5:2) the diazirine 30 was obtained (404 mg, 0.60 mmol, 49%) as a
colorless solid. [α]2D0 = +5.7 (c = 1.15, CHCl3). 1H NMR (500 MHz,
[1] a) Y. Hatanaka, H. Nakayama, Y. Kanaoka, Rev. Heteroat.
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3
3
CDCl3): δ = 4.97 (d, J1,2 Ϸ 0 Hz, 2 H, 2 H-1), 4.13 (ddϷd, J2,3
3
= 5.7 Hz, 2 H, 2 H-2), 4.08 (dd, J2,3 = 7.9 Hz, 2 H, 2 H-3), 3.81
2
(dd, J5,6 = 5.6 Hz, 2 H, 2 H-6), 3.74–3.64 (m, 6 H, 2 H-4, 2 H-6Ј,
3J2,3 = 10.8 Hz, 2 Ha), 3.57–3.48 (m, 8 H, 2 H-5, 2 Hc,cЈ, 2 HaЈ),
3.39 (d, 2 H, 2 Hb), 3.33 (d, 2 H, 2 HbЈ), 1.48, 1.45, 1.35, 1.28 (each
s, each 6 H, 8 iPr-CH3) ppm. 13C NMR (125.75, CDCl3): δ =
109.44 (2 2,3-iPr-Cq), 99.69 (2 4,6-iPr-Cq), 97.80 (2 C-1), 75.96 (2
C-2), 74.81 (2 C-3), 72.70 (2 C-4), 70.29 (2 Cb), 70.01 (2 Cc), 66.47
(2 Ca), 62.03 (2 C-6), 61.39 (2 C-5), 29.05, 28.17, 26.15, 18.78 (8
iPr-CH3), 27.38 [C(NN)] ppm. MALDI-TOF MS (norharmane):
m/z = 669.79 [M + Na – N2 ]+ (calcd. m/z = 669.31) for
C31H50NaO15.
1,9-Bis(α-D-mannopyranosyloxy)-5-azi-3,7-dioxanonane (31): The
protected compound 30 (187 mg, 0.28 mmol) was dissolved in a 9:1
mixture of TFA and water (10 mL). After 1 min the solution was
concentrated in vacuo at room temp. The residue was codistilled
twice with toluene and after column chromatography (ethyl acetate/
methanol, 1:2) the title compound (133 mg, 0.26 mg, 93%) was de-
livered as a colorless solid. [α]2D0 = +57.1 (c = 0.95, MeOH). 1H
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3
NMR (500 MHz, CD3OD): δ = 4.84 (d, J1,2 = 1.4 Hz, 2 H, H-1),
3.90–3.82 (m, 3J2,3 = 3.3, 3J5,6Ј = 2.4 Hz, 6 H, H-2, H-6Ј, Ha), 3.79–
3.72 (m, 3J5,6 = 5.9, 2J6,6Ј = 11.7 Hz, 4 H, H-3, H-6), 3.69–3.57 (m,
10 H, H-4, H-5, HaЈ, Hc,cЈ), 3.47 (d, 4 H, Hb,bЈ) ppm. 13C NMR
(125.75, CD3OD): δ = 101.70 (C-1), 74.60 (C-5), 72.55 (C-3), 72.09
(C-2), 71.33 (Cc), 71.13 (Cb), 68.57 (C-4), 67.63 (Ca), 62.88 (C-6),
28.69 [C(NN)] ppm. MALDI-TOF MS (DHB): m/z = 509.54 [M
+ Na – N2]+ (calcd. m/z = 509.19) for C19H34 NaO14, 537.52 [M +
Na]+ (calcd. m/z = 537.19) for C19H34 N2NaO14. ESI MS: m/z =
537.2150 [M + Na]+ (calcd. m/z = 537.1902) for C19H34N2NaO14.
M. Kuldová, P. Rossmann, O. Horváth, V. Kren, P. Krist, K.
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Acknowledgments
This work was supported by the Deutsche Forschungsgemeinschaft
(DFG) in the frame of SFB470.
Received: September 9, 2005
Published Online: November 21, 2005
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