Synthesis of mono- and di-α-l-fucosylated 2-acetamido-2-deoxy-N-glycyl-β-d-glucopyranosylamines, spacered fragments of glycans of N-glycoproteins

Article abstract of DOI:10.1007/s11172-015-0990-7

α-l-Fucp-(1→3)-d-GlcNAc, α-l-Fucp-(1→6)-[α-l-Fucp-(1→3)]-d-GlcNAc, and β-d-Galp-(1→3)-[α-l-Fucp-(1→4)]-d-GlcNAc were converted into corresponding β-glycopyranosylamines by action of ammonium carbamate in aqueous boric acid, or in aqueous methanol in case

Full text of DOI:10.1007/s11172-015-0990-7

Russian Chemical Bulletin, International Edition, Vol. 64, No. 5, pp. 1134—1141, May, 2015
1134
Synthesis of monoꢀ and diꢀꢀLꢀfucosylated 2ꢀacetamidoꢀ2ꢀdeoxyꢀ
NꢀglycylꢀꢀDꢀglucopyranosylamines, spacered fragments
of glycans of Nꢀglycoproteins*
L. M. Likhosherstov,a O. S. Novikova,a N. N. Malysheva,a and V. E. Piskarevb
аN. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences,
47 Leninsky prosp., 119991 Moscow, Russian Federation.
Fax: (495) 135 5328. Eꢀmail: likhosherstov@mail.ru
bA. N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences,
28 ul. Vavilova, 119991 Moscow, Russian Federation
ꢀLꢀFucpꢀ(13)ꢀDꢀGlcNAc, ꢀLꢀFucpꢀ(16)ꢀ[ꢀLꢀFucpꢀ(13)]ꢀDꢀGlcNAc, and ꢀDꢀ
Galpꢀ(13)ꢀ[ꢀLꢀFucpꢀ(14)]ꢀDꢀGlcNAc were converted into corresponding ꢀglycopyraꢀ
nosylamines by action of ammonium carbamate in aqueous boric acid, or in aqueous methanol
in case of ꢀLꢀFucpꢀ(16)ꢀDꢀGlcNAc. NꢀAcylation of these fucooligosaccharides with
NꢀZꢀglycine Nꢀhydroxysuccinimide ester (Z is benzyloxycarbonyl) followed by hydrogenolytic
removal of Zꢀgroup afforded corresponding Nꢀglycylꢀꢀglycopyranosylamines of these fucooliꢀ
gosaccharides; three of them model carbohydrateꢀpeptide region of Nꢀglycoproteins, and the
forth is an aminoꢀspacered Leaꢀantigen.
Key words: ꢀLꢀfucose, 2ꢀacetamidoꢀ2ꢀdeoxyꢀꢀDꢀglucopyranose, oligosaccharides, Nꢀglyꢀ
cylꢀꢀglucopyranosylamines, Nꢀacylation, carbohydrateꢀpeptide bond, Nꢀglycoproteins.
ꢀLꢀFucopyranose (Fuc) is an important component
they are, all of them are linked to the polypeptide chains
through the DꢀGlcNAc1ꢀNꢀAsn fragment. In some types
of Nꢀglycans the GlcNAc (GlcNAcꢀ1) residue is modiꢀ
fied with Fuc at 6ꢀ or 3ꢀpositions, and in some cases at
both of them. The first type of modification was found in
many animals, for example, mammals and birds, the secꢀ
ond type was observed in plants, insects, helminths, and
slime mold, and the third type was found in insects and
some helminths.8—10 NꢀGlycans, which comprise fucoꢀ
sylated GlcNAcꢀ1, show pronounced biological activity.
For example, it was shown, that the presence of ꢀ1ꢀ6ꢀ
fucose residue decreases cytotoxicity of Nꢀglycoproteins,11
thus providing for monitoring of the efficacy of theraꢀ
peutic antibodies of Gꢀtype.12 ꢀ1ꢀ3ꢀFucosylated Nꢀglyꢀ
cans are highly immunogenic for mammals and are one of
the causes of allergic reactions to food from plant sources
and to insect stings in humans.13
In the present work we report the application of
the carbamate involving procedure for the synthesis of
ꢀglycopyranosylamines,14a,b which was elaborated in our
group earlier, to alkali labile fucooligosaccharides with
GlcNAc on the reducing end followed by preparation
of Nꢀglycylꢀꢀglycopyranosidic derivatives thereof.
Diꢀ and trisaccharides ꢀLꢀFucpꢀ(13)ꢀDꢀGlcNAc (1a),
ꢀLꢀFucpꢀ(13)ꢀ[ꢀLꢀFucpꢀ(16)]ꢀDꢀGlcNAc (2a),
ꢀDꢀGalpꢀ(13)ꢀ[ꢀLꢀFucpꢀ(14)]ꢀDꢀGlcNAc (3a) and
ꢀLꢀFucpꢀ(16)ꢀDꢀGlcNAc (4a), which are the fragments
of many glycoconjugates, and in particular, glycoproteins
and glycolipids. In these biopolymers, as a rule, Fuc is
linked to 2ꢀnd position of ꢀDꢀgalactopyranose (Gal) or
to 3ꢀ, 4ꢀ or 6ꢀth positions of 2ꢀacetamidoꢀ2ꢀdeoxyꢀꢀDꢀ
glucopyranose (GlcNAc) (see Ref. 1) as a terminal moiety.
Syntheses of fucooligosaccharides as free oligosaccharides,
Оꢀglycosides or Оꢀspacered derivatives are described in
the literature (see, for example, review2). Data on Nꢀspacꢀ
ered fucooligosaccharides is not so plentiful2,3а and it is
often referred to Nꢀasparagine derivatives.3b One type of
Nꢀspacered oligosaccharides used for preparation of conꢀ
jugates are Nꢀglycylꢀꢀglycopyranosylamines (see Ref. 4
and references cited therein). Earlier, we reported syntheꢀ
sis of derivatives of this type from oligosaccharides of breast
milk.4,5 Some of these compounds were used in our reꢀ
search6 aimed at natural inhibitors of two norovirus strains,
which belong to the Caliciviridae family of RNA viruses,
which are the leading cause of epidemics of acute gastroꢀ
enteritis worldwide.7
Biosynthesis of Nꢀglycoproteins in eukaryotic cells can
be accompanied by formation of considerable quantities
of Nꢀglycans, which differ in monosaccharide composiꢀ
tion and molecular weight; however, being different as
* On the occasion of the 100th anniversary of the birth of Acadeꢀ
mician N. K. Kochetkov (1915—2005).
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 5, pp. 1134—1141, May, 2015.
1066ꢀ5285/15/6405ꢀ1134 © 2015 Springer Science+Business Media, Inc.

NꢀGlycylꢀglycosylamines of fucosaccharides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 5, May, 2015
1135
of physiologically important natural glycoconjugates, were
used in this research. Disaccharide 1a is a fragment of the
aforementioned carbohydrateꢀpeptide linking unit in
Nꢀglycoproteins of plants, insects, helminths, and slime
mold.9,10 Besides, it is a part of carcinoembryonic antigen
Lex and its derivatives (SiaLex, SulfoLex, VIMꢀ2, LDNF),
which were found in many glycoconjugates. Trisaccharide
2a is a part of a carbohydrate—peptide linking unit in
Nꢀglycoproteins of insects and some helminths.9,10 Trisacꢀ
charide 3a is a carcinoembryonic antigen Lea, which was
found in many animal glycoconjugates. As a part of comꢀ
plex Nꢀglycans in plant glycoproteins, Lea is an antigen
determinant, and it is highly immunogenic for animals,
being one of the causes of allergic reaction to plants in
humans.13 Disaccharide 4a is a part of a carbohydrate—
peptide linking unit in Nꢀglycoproteins found in many
species of animals, including mammals and humans.8
Syntheses of oligosaccharides 1a,15 3a,16 and 4a17 were
described earlier, and trisaccharide 2a was synthesized as
an allyl Oꢀglycoside18 and a part of tetraꢀ and octasacchaꢀ
rides.19 Syntheses of oligosaccharides 1a—4a were perꢀ
formed by our group (Schemes 1 and 2) with the use of
earlier described approaches and procedures except that
we chose to use ethyl 2,3,4ꢀtriꢀOꢀbenzylꢀ1ꢀthioꢀꢀLꢀfucoꢀ
pyranoside (5) in the presence of Et4NBr and CuBr220 for
fucosylation. Herewith we have found, that increased
quantity of Et4NBr (3.5 molar excess instead of 1.5) reꢀ
sults in considerable improvement of stereoselectivity of
Scheme 1
Reagents and conditions: a. Et4NBr, CuBr2, CH2Cl2, DMF, ~20 C, 64 h; b. 90% aqueous TFA, CHCl3, 27 C, 30 min; c. MeONa,
MeOH, ~20 C, 5 h.

1136
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 5, May, 2015
Likhosherstov et al.
ꢀLꢀfucosylation ( :  ratio of anomers was ~9 : 1 inꢀ
stead of ~4 : 1). It should be noted, that we performed
hydrogenolytic removal of benzyl groups (with Pd/C
as a catalyst) at higher temperature, namely, ~50 C,
without addition of acetic acid or increasing the presꢀ
sure of H2.
On the basis of literature data we assumed that oligoꢀ
saccharides 1a—4a may have different resistance to alkali.
It is known21 that 2ꢀacetamidoꢀ2ꢀdeoxyhexoses are more
alkaliꢀlabile than the corresponding hexoses. The presꢀ
ence of a monosaccharide residue at position 6 of 2ꢀacetꢀ
amidoꢀ2ꢀdeoxyhexoses has no effect on its resistance to
alkaline conditions, and the presence of the substituent at
position 4 considerably enhances the resistance of the
oligosaccharide. Substituent at position 3 dramatically deꢀ
creases the stability of 2ꢀacetamidoꢀ2ꢀdeoxyhexose faciliꢀ
tating ꢀelimination of the substituent and formation of so
called chromogens21 along with 3,6ꢀanhydroꢀ2ꢀacetꢀ
amidoꢀ2ꢀdeoxyhexoses.22
The conditions, which we reported7 earlier for the synꢀ
thesis of ꢀglycopyranosylamines of alkaliꢀlabile oligosacꢀ
charides with a glucose residue on the reducing end (satuꢀ
rated solution of ammonium carbamate in ~90% aqueous
MeOH, ~20 C, 100 h), were found to be efficient only for
disaccharide 4a. Under these conditions, oligosaccharides
1a—3a were found to decompose, especially oligosacchaꢀ
rides 1a and 2a. According to data of electrophoresis analꢀ
ysis, the reaction conditions result in transformation of
oligosaccharides 1a, 2a or 3a into one neutral and at least
two positively charged products. The neutral product may
be a mixture of the starting oligosaccharide and decompoꢀ
sition products which are disaccharide 1a and 3,6ꢀanꢀ
hydroꢀ2ꢀacetamidoꢀ2ꢀdeoxyꢀDꢀhexofuranose,22 together
with oligosaccharides 1a—3a and corresponding chroꢀ
mogens.21 Two positively charged products have mobility,
which is close to both ꢀglycopyranosylamines of diꢀ and
trisaccharides (ꢀglycopyranosylamines of lactose and
2´ꢀfucosyllactose were used as reference compounds) and
to ꢀglycopyranosylamines of monosaccharides (ꢀgalacꢀ
topyranosylamine was used as a reference compound). Preꢀ
sumably, the former are ꢀglycopyranosylamines of oliꢀ
gosaccharides 1b—3b, and the latter are ꢀglycopyranoꢀ
sylamines of fucose or galactose, which are formed as
a result of ꢀelimination of oligosaccharides 1a—2a or
trisaccharide 3a, respectively.
It is known,24 that H3BO3 and its salts form negatively
charged complexes with sugars. Presumably, formation of
these complexes slows down ꢀelimination of fucose and
galactose residues from the position 3 of oligosaccharides
1a—3a (the effect was especially pronounced in case of
oligosaccharide 1a), and results in the increase of yields of
ꢀglycopyranosylamines 1b—3b. ꢀGlycopyranosylamines
1b—4b were not isolated from the reaction mixtures as
pure compounds as they are labile and can easily undergo
different transformations. ꢀGlycopyranosylamines were
transformed into stable Nꢀacylated derivatives without
purification using NꢀZꢀglycine Nꢀhydroxysuccinimide ester
as described in Ref. 7 (see Scheme 2). NꢀAcylꢀꢀglycoꢀ
pyranosylamines 1c—4c were isolated using chromatoꢀ
graphy on reversed phase silica gel (for 1c, 3c, and 4c),
and for 2c, additionally, adsorption chromatography on
silica gel was used. Compounds 1c, 2c, 3c, and 4c were
obtained with yields 47, 17, 64, and 72%, respectively. It
should be mentioned, that although boric acid was added
to the saturated aqueous solution of ammonium carbaꢀ
mate, preparation of ꢀglycosylamine 2b from 3,6ꢀdifuꢀ
Scheme 2
In connection with the obtained data, the research
work was carried out aimed at optimization of procedure
for preparation of ꢀglycopyranosylamines, which decreasꢀ
es decomposition of oligosaccharides 1a—3a. It was found,
that the use of ammonium bicarbonate23 instead of amꢀ
monium carbamate decreased the formation of decompoꢀ
sition products only to a small extent. Further investigaꢀ
tion showed that addition of H3BO3 to the saturated aqueꢀ
ous ammonium carbamate decreases the decomposition
of oligosaccharides 1a—3a.
R = OH (1a—4a), NH2 (1b—4b), NHCOCH2NHZ (1c—4c),
NHCOCH2NH2 (1d—4d)
Z = PhCH2OCO
Reagents and conditions: a. H2, Pd/C, 50 C, EtOH, 48 h;
b. NH2COONH4, H3BO3 (for 1b—3b), H2O or MeOH + H2O
(for 4a), ~20 C, 96 h; c. ZNHCH2COOSu (Su — succinimidyl),
DMF, H2O, 0 C, 3 h; d. H2, Pd/C, MeOH, H2O, ~20 C, 7 h.

NꢀGlycylꢀglycosylamines of fucosaccharides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 5, May, 2015
1137
Experimental
cosylated GlcNAc 2a was accompanied by substantial deꢀ
composition and the yield of Nꢀacylated derivative 2c is
still low.
1H and 13C NMR spectra were recorded in D2O on a Bruker
AM 300 spectrometer (operating frequencies 300.13 and
75.47 MHz) relative to the signals of acetone H 2.225, C 31.45
(internal standard). Only characteristic proton signals are given.
High resolution mass spectra were recorded on Bruker
micrOTOF II mass spectrometer with electrospray ionization.15
The mass spectra were registered in the positiveꢀ and negativeꢀ
ion modes (capillary voltage was –4500 and 3200 V, respectꢀ
ively). Experimental m/z values are given for the most abundant
signals of isotopic clusters together with the corresponding calꢀ
culated m/z values. Solutions of samples in aqueous methanol or
in MeOH—H2O (1 : 1), which were diluted with MeCN or
MeCN—H2O (1 : 1), were introduced by the syringe injection.
Optical rotation was measured on polarimeter PUꢀ07 (Russia).
Electrophoresis (10 V cm–1, 1 h) was carried out on a Filtrak
FN1 paper in 4% aqueous HCOOH solution, using EFAꢀ1 inꢀ
strument (USSR). The spots of compounds were visualized
with the sequence of reagents KIO4—AgNO3—KOH (see Ref. 16).
Molecular sieves were activated in vacuo at ~0.5 Torr, 200 C, 2 h.
Solutions were concentrated at ~10 Torr, bath temperature was
~30 °С. Detection of all compounds during chromatography on
Silica gel 100 C18 Reversed phase (Fluka) was carried out using
UVꢀabsorption at 206 nm.
After removal of protective groups by hydrogenolysis
of 1c—4c using the procedure earlier developed by our
group,7 Nꢀglycyl derivatives 1d—4d were obtained with
yields ~95%.
Structures of the obtained compounds 1a,b,c,d—
4a,b,c,d, 7, 8, 11, 13, 14 were confirmed using the high
resolution massꢀ and 1H and 13C NMR spectrometry (for
compounds 2a,c,d).
In this way, the procedure suggested14 by our group
earlier, which assumes the use of ammonium carbamate
for preparation of ꢀglycopyranosylamines of monoꢀ and
oligosaccharides, after modification (addition of H3BO3)
can be applied to the synthesis of ꢀglycopyranosylamines
1b—3b of known alkaliꢀlabile oligosaccharides 1a—3a with
Fuc or Gal at position 3 of reducing GlcNAc. The develꢀ
oped earlier procedure5 of Nꢀacylation of ꢀglycopyranoꢀ
sylamines 1b—4b with activated ester of NꢀZꢀglycine proꢀ
vided for Nꢀacylated compounds 1c—4c. Removal of the
protective group afforded Nꢀglycylꢀꢀglycopyranosylꢀ
amines of four fucooligosaccharides; three of them (1d,
2d, and 4d) are models of carbohydrate—peptide linking
unit in Nꢀglycoproteins, and 3d is an amino spacered deꢀ
terminant of carcinoembryonic antigen Lea.
Aminoꢀspacered compounds 1d—4d can be used for
immobilization on different carriers for different types of
ELISA and for affinity chromatography of fucolectins.
The primary amino group of glycine provides for formaꢀ
tion of conjugates with different physiologically active
compounds, (bio)polymers or nanoparticles in mild conꢀ
ditions. Glycoconjugates of this type can be used in differꢀ
ent test systems, for example, for investigation of specificꢀ
ity of fucolectins with affinity to core Fuc, one of them is
ꢀLꢀFucpꢀ(16)ꢀDꢀGlcNAcꢀspecific lectin from Aspergilꢀ
lus oryzae,25 and also for study of Fucꢀspecific adhesins of
bacteria and viruses.
Fragments of ꢀ1ꢀ3ꢀfucosylated Nꢀglycans can be used
as immunogenic ligands for formulation of synthetic vacꢀ
cines for treatment of helminthoses.26 It was shown, that
vaccination of sheep with glycoprotein Н(11) isolated from
nematodes Haemonchus contortus, which cause parasitic
infections in some domestic animals, mounted immune
response to other sheep parasites (see review27). Glycoꢀ
proteins H. contortus include considerable amount of core
antigen structures ꢀLꢀFucpꢀ(13)ꢀDꢀGlcNAc (1a) and
ꢀLꢀFucpꢀ(13)ꢀ[ꢀLꢀFucpꢀ(16)]ꢀDꢀGlcNAc (2а), and
the immune response caused by the vaccination is basicalꢀ
ly connected to the presence of these motifs.28 In this
aspect, aminoꢀsynthons 1d and 2d can be conjugated to
a polymer carrier (protein, waterꢀsoluble protein, or a nanoꢀ
particle) with the view to application as potential candiꢀ
dates for vaccination of mammals and humans against
parasitic worms nematodes and trematodes.
Benzyl 2ꢀacetamidoꢀ3ꢀОꢀ(2,3,4ꢀtriꢀОꢀbenzylꢀꢀLꢀfucopyraꢀ
nosyl)ꢀ2ꢀdeoxyꢀꢀDꢀglucopyranoside (7). Ethyl 2,3,4ꢀtriꢀОꢀbenzꢀ
ylꢀ1ꢀthioꢀꢀLꢀfucopyranoside31 (5) (956 mg, 2 mmol), Et4NBr
(1.470 g, 7 mmole), CuBr2 (535 mg, 2.4 mmol) and powdered
molecular sieves 3 Å (2.4 g) in the mixture of anhydrous CH2Cl2
(20 ml) and DMF (4 ml) were stirred for 4 h at 21 С. Benzyl
2ꢀacetamidoꢀ4,6ꢀОꢀbenzylideneꢀ2ꢀdeoxyꢀꢀDꢀglucopyranoꢀ
side32 (6) (800 mg, 2 mmol) was added to the suspension, and it
was stirred for 64 h at 21 С. The reaction mixture was diluted
with CHCl3 (125 mL), filtered through Celite, and Celite was
washed with CHCl3 until filtrate became colorless. The solution
was concentrated to ~120 mL and washed with 1 M aqueous
Na2S2O3 until the color diappeared, then washed with H2O
(330 mL) and the solvent was removed. The residue was disꢀ
solved in benzene and subjected to chromatography on silica gel
(benzene  benzene—acetone (7 : 3)). Fractions were concenꢀ
trated until dryness and analyzed with TLC using benzene—
acetone (7 : 3) as an eluent. Fractions with nearly homogenious
product were combined to yield 881 mg of benzyl 2ꢀacetamidoꢀ
4,6ꢀОꢀbenzylideneꢀ3ꢀОꢀ(2,3,4ꢀtriꢀОꢀbenzylꢀꢀLꢀfucopyranoꢀ
syl)ꢀ2ꢀdeoxyꢀꢀDꢀglucopyranoside which was without further
purification dissolved in 6 mL of CHCl3, then 0.9 mL of 90%
aqueous TFA was added and the mixture was kept for 30 min at
27 С. The reaction mixture was diluted with 6 mL of CHCl3,
cooled with an ice bath and added 1.8 mL of Et3N and 20 mL of
PriOH. The solution was concentrated until dryness. The resiꢀ
due was triturated with petroleum ether, dissolved in 15 mL of
CHCl3. The solution was washed with H2O (52 mL), diluted
with 10 mL of PriOH and concentrated to dryness. The residue
was subjected to column chromatography on silica gel using
CH2Cl2—Et2O (1 : 1) as an eluent. Fractions with homogeneous
product were combined and the solvents were removed to obꢀ
tain 653 mg of disaccharide 7 (yield 45%) which can be crystalꢀ
21
lized from MeOH, m.p. 204—205 С (needles), []D +26.5
(с 1, CНCl3); compared to Ref.15a data: m.p. 178—180 С,
25
[]D +26.4 (с 1, CНCl3). MS (positive ions), m/z: 728.3415,

1138
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 5, May, 2015
Likhosherstov et al.
750.3235, 766.2973, С42Н49NО10, calc., : 728.3429 [M + H]+,
750.3249 [M + Na]+, 766.2988 [M + K]+. 1H NMR,  (CDCl3):
1.17 (d, 3 H, CH3, Fuc, J = 6.5 Hz); 1.40 (s, 3 H, NAc); 5.23
(d, 1 H, H(1), GlcNAc, J = 3.3 Hz); 5.99 (d, 1 H, NH, J = 7.6 Hz);
7.16—7.48 (m, 20 H, 4 Ph).
Benzyl 2ꢀacetamidoꢀ3,6ꢀdiꢀOꢀ(2,3,4ꢀtriꢀOꢀbenzylꢀꢀLꢀfucoꢀ
pyranosyl)ꢀ2ꢀdeoxyꢀꢀDꢀglucopyranoside (8) was obtained from
disaccharide 7 (291 mg, 0.4 mmol) and thioglycoside 5 (191 mg,
0.4 mmol) as described for disaccharide 7. Chromatography on
silica gel (benzene benzene—acetone (8 : 2)) gave 247 mg
(54%) of amorphous trisaccharide 8, []D20 –18.8 (с 1, CНCl3).
MS (positive ions), m/z: 1166.5232, С69Н77NО14, calc.
1166.5236 [M + Na]+. 1H NMR, (CDCl3): 1.04 (d, 3 H, CH3,
Fuc13, J = 6.5 Hz); 1.17 (d, 3 H, CH3, Fuc16, J = 6.5 Hz);
1.46 (s, 3 H, NAc); 3.56 (br.s, 1 H, H(4), Fuc13); 5.22 (d, 1 H,
H(1), GlcNAc, J = 3.0 Hz); 5.99 (d, 1 H, NH, J = 6.6 Hz);
7.19—7.46 (m, 35 H, 7 Ph).
the solution was kept for 5 h at 20 С. Then the solution was
diluted with 4 mL of MeOH, 75 mg of Amberlyst 15 (H+) was
added, the reaction mixture was stirred for 1 h, the resin was
filtered off and washed with MeOH. The solution was concenꢀ
trated to dryness. The residue was subjected to column chromaꢀ
tography on silica gel in CH2Cl2  CH2Cl2—Et2O (1 : 1). Fracꢀ
tion with homogeneous product were combined and concentratꢀ
ed to dryness to obtain 113 mg (78%) of amorphous disaccharide
14, []D21 +6.5 (с 1, CНCl3). MS (positive ions), m/z: 728.3441,
750.3254, 766.2994, С42Н49NО10, calc., 728.3429 [M + H]+;
750.3249 [M + Na]+; 766.2988 [M + K]+. 1H NMR,  (CDCl3):
1.13 (d, 3 H, CH3, Fuc, J = 6.4 Hz); 2.00 (s, 3 H, NAc); 5.78
(d, 1 H, NH, J = 8.2 Hz); 7.23—7.45 (m, 20 H, 4 Ph).
Hydrogenolysis of 7, 8, 11 и 14 (general procedure). The
suspension of 10% Pd/C (75 mg, the catalyst) in 1.5 mL of EtOH
was stirred under H2 (30 min at ~50 С). The solution of comꢀ
pounds 7, 8, 11 or 14 (150 mg) in 12 mL of EtOH was heated to
~50 С and added to the suspension at ~50 С, and hydrogenaꢀ
tion was performed upon stirring under a weak flow of H2 for
24 h at ~50 С. Then 1.5 mL of H2O and 75 mg of 10% Pd/C was
added to the suspension, and the hydrogenation was conducted
for another 24 h at ~50 С. The reaction mixture was diluted
with 8 mL of 50% aqueous MeOH, the catalyst was removed by
centrifugation, washed with 50% aqueous MeOH and the soluꢀ
tion was concentrated to dryness. The residue was dissolved in
1 mL of H2O and applied onto the C18ꢀreversed phase silica gel
column (0.6×4.5 см). The product was eluted with 30 mL of
H2O, the solution was concentrated and dried. Oligosacchaꢀ
rides 1a—4a were obtained with yield ~95%.
Benzyl 2ꢀacetamidoꢀ6ꢀOꢀbenzylꢀ4ꢀOꢀ(2,3,4ꢀtriꢀOꢀbenzylꢀꢀ
Lꢀfucopyranosyl)ꢀ3ꢀOꢀ(ꢀDꢀgalactopyranosyl)ꢀ2ꢀdeoxyꢀꢀDꢀglucoꢀ
pyranoside (11) was obtained from benzyl 2ꢀacetamidoꢀ6ꢀOꢀ
benzylꢀ3ꢀOꢀ(2,3,4,6ꢀtetraꢀOꢀacetylꢀꢀDꢀgalactopyranosyl)ꢀ2ꢀ
deoxyꢀꢀDꢀglucopyranoside16a (9) (292.5 mg, 0.4 mmol) and
ethyl 2,3,4ꢀtriꢀOꢀbenzylꢀ1ꢀthioꢀꢀLꢀfucopyranoside31 (5) (191 mg,
0.4 mmol) as described for disaccharide 7. After chromatography
on silica gel (ethyl acetate—hexane (1 : 1)  ethyl acetate—
hexane (3 : 1)) 238.6 mg (52%) of benzyl 2ꢀacetamidoꢀ3ꢀOꢀ
(2,3,4,6ꢀtetraꢀOꢀacetylꢀꢀDꢀgalactopyranosyl)ꢀ4ꢀOꢀ(2,3,4ꢀtriꢀOꢀ
benzylꢀꢀLꢀfucopyranosyl)ꢀ6ꢀOꢀbenzylꢀ2ꢀdeoxyꢀꢀDꢀglucopyrꢀ
anoside16a (10) was obtained, which was dissolved in 4 mL of
anhydrous MeOH and 0.4 mL of 1 М MeONa in anhydrous
MeOH was added. The solution was kept for 5 h at 20 С. Then
the solution was diluted with 4 mL of MeOH, 75 mg of Amꢀ
berlyst 15 (H+) was added, the reaction mixture was stirred for 1 h,
the resin was filtered off and washed with MeOH. The solution
was concentrated to dryness to obtain 170 mg (82%) of trisacꢀ
charide 11, which is crystallized from EtOH, m.p. 189—191 С,
[]D26 –14.3 (с 1, CНCl3); compare to Ref. 16a: m.p. 180—181 С
from the mixture ethyl acetate—ether, []D –15 (с 1, CНCl3).
MS (positive ions), m/z: 980.4421, 1002.4251, 1018.3987,
С55Н65NО15, calc. 980.4427 [M + H]+; 1002.4246 [M + Na]+;
1018.3986 [M + K]+. MS (negative ions), m/z: 978.4261.
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ3ꢀOꢀ(ꢀLꢀfucopyranosyl)ꢀDꢀglucose
(1a), m.p. 218—220 С from EtOH, []D22 –84.9 (after 15 min)
 –102.2 (after 24 h) (с 1, Н2О); compare with Ref.: m.p.
20
218—220 С from EtOH—MeOH—H2O, []D –60  –74 (с
0.83, Н2О)15a; []D22 –66.615b. MS (positive ions), m/z: 368.1554,
390.1372, 406.1001, С14Н25NO10, calc., 368.1551 [M + H]+;
390.1371 [M + Na]+; 406.1110 [M + K]+. 1H NMR, 1.18 (d, 3 H,
CH3, Fuc, J = 6.5 Hz); 2.04 (s, 3 H, NAc); 4.35 (m, 1 H, H(5),
Fuc); 4.73 (d, H(1), ꢀGlcNAc, J = 8.3 Hz); 5.00 and 5.03 (both d,
H(1), Fuc, J = 3.9 Hz); 5.15 (d, H(1), ꢀGlcNAc, J = 3.4 Hz).
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ3,6ꢀdiꢀOꢀ(ꢀLꢀfucopyranosyl)ꢀDꢀgluꢀ
21
cose (2a), []D –93.4 (с 1, Н2О). MS (positive ions), m/z:
1
1
С55Н65NO15, calc. 978.4281 [M – H]–. H NMR, (CDCl3):
536.1941, С20H35NO14, calc., 536.1950 [M + Na]+. H NMR
1.12 (d, 3 H, CH3, Fuc, J = 6.5 Hz); 1.95 (s, 3 H, NAc); 5.09 (d,
1 H, H(1), GlcNAc, J = 3.2 Hz); 5.84 (d, 1 H, NH, J = 7.7 Hz);
7.20—7.46 (m, 25 H, 5 Ph).
,: 1.17 (m, 3 H, CH3, Fuc13); 1.22 (m, 3 H, CH3, Fuc16);
2.02 (s, 3 H, NAc); 4.12 (m, 1 H, H(5), Fuc16); 4.33 (m, 1 H,
H(5), Fuc1 3); 4.73 (d, H(1), ꢀGlcNAc, J = 8.2 Hz); 4.92
(m, 1 H, H(1), Fuc16); 4.98, 5.02 (both d, H(1), Fuc13); 5.14
(d, H(1), ꢀGlcNAc, J = 3.4 Hz). 13C NMR, : 16.49 (2 C(6),
2 Fuc); 23.23 (CH3CO, ꢀGlcNAc); 23.48 (CH3CO, ꢀGlcNAc);
54.84 (C(2), ꢀGlcNAc); 57.63 (C(2), ꢀGlcNAc); 67.97 (C(5),
Fuc16); 68.14 (C(5), Fuc13); 68.59 (C(6), ꢀGlcNAc);
68.97(C(6), ꢀGlcNAc); 69.24 (C(2), Fuc13); 69.47 (C(2),
Fuc16); 69.89 (C(4), GlcNAc); 70.79 (2 C(3), 2 Fuc); 72.25
(C(5), ꢀGlcNAc); 73.08 (2 C(4), 2 Fuc); 76.24 (C(5),
ꢀGlcNAc); 79.27 (C(3), ꢀGlcNAc); 81.75 (C(3), ꢀGlcNAc);
92.29 (C(1), ꢀGlcNAc); 96.01 (C(1), ꢀGlcNAc); 100.32,
100.67 (C(1), Fuc16); 101.01, 101.24 (C(1), Fuc13); 175.75
(C=O); 176.02 (C=O).
Benzyl 2ꢀacetamidoꢀ3,4ꢀdiꢀOꢀacetylꢀ6ꢀOꢀ(2,3,4ꢀtriꢀOꢀbenzꢀ
ylꢀꢀLꢀfucopyranosyl)ꢀ2ꢀdeoxyꢀꢀDꢀglucopyranoside (13) was
obtained from benzyl 2ꢀacetamidoꢀ3,4ꢀdiꢀOꢀacetylꢀ2ꢀdeoxyꢀꢀ
Dꢀglucopyranoside17b (12) (158 mg, 0.4 mmol) and thioglycoꢀ
side 5 (191 mg, 0.4 mmol) as desribed for disaccharide 7. Chroꢀ
matography on silica gel (benzene benzene—acetone (8 : 2))
21
gave 175 mg (54%) of amorphous disaccharide 6, []D +24.1
(с 1, CНCl3). MS (positive ions), m/z: 834.3473, 850.3211,
С46Н53NО12, calc., 834.3460 [M + Na]+; 850.3199 [M + K]+.
1H NMR,  (CDCl3): 1.17 (d, 3 H, CH3, Fuc, J = 6.5 Hz); 1.95
(s, 3 H, NAc); 2.03 (s, 3 H, Ac); 2.07 (s, 3 H, Ac); 5.70 (d, 1 H,
NH, J = 9.4 Hz); 7.20—7.40 (m, 20 H, 4 Ph).
Benzyl 2ꢀacetamidoꢀ6ꢀOꢀ(2,3,4ꢀtriꢀOꢀbenzylꢀꢀLꢀfucopyraꢀ
nosyl)ꢀ2ꢀdeoxyꢀꢀDꢀglucopyranoside (14). Disaccharide 13
(162 mg, 0.2 mmol) was dissolved in 4 mL of anhydrous MeOH
and 0.4 mL of 1 М MeONa in anhydrous MeOH was added, and
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ4ꢀOꢀ(ꢀLꢀfucopyranosyl)ꢀ3ꢀOꢀ(ꢀDꢀ
galactopyranosyl)ꢀDꢀglucose (3a), m.p. 168—170 С from EtOH,
21
[]D –48.2 (after 15 min)  –53.9 (after 24 h) (с 1, Н2О);
compare to Ref.: []D –44.5 (с 1, Н2О)16a,c; []D –45.1
25

NꢀGlycylꢀglycosylamines of fucosaccharides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 5, May, 2015
1139
(с 1, Н2О)16b. MS (positive ions), m/z: 552.1890, С20Н35NO15,
calc., 552.1899 [M + Na]+. 1H NMR, : 1.18 (d, 3 H, CH3, Fuc,
J = 6.5 Hz); 2.02 (s, 3 H, NAc); 4.47 and 4.50 (both d, H(1),
Gal, J = 7.5 Hz); 4.71 (d, H(1), ꢀGlcNAc, J = 8.2 Hz); 4.88
(m, 1 H, H(5), Fuc); 5.02 (m, 1 H, H(1), Fuc); 5.12 (d, H(1),
ꢀGlcNAc, J = 3.2 Hz).
subjected to column chromatography on С18ꢀreversed phase silica
gel in the described conditions. The methanol—water fractions,
which contained homogeneous compound (TLC, EtOAc—
MeOH (2.5 : 1)), were combined and concentrated to dryness.
The residue was dried to yield 26 mg (47%) of compound 1c,
46 mg (64%) of compound 3c, 40.2 mg (72%) of compound 4с.
Compound 2c (16 mg) was additionally chromatographed on
silica gel (acetone—MeOH (9 : 1)  acetone—MeOH (4 : 1)) to
obtain 12 mg (17%) of compound 2c.
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ6ꢀOꢀ(ꢀLꢀfucopyranosyl)ꢀDꢀglucose
22
25
(4a), []D –61.2 (с 1, Н2О); compare with Ref.: []D –65
(с 1.05, Н2О)17a,b; []D –61.2  –63.2 (с 1, Н2О)17c. MS (positive
ions), m/z: 368.1550, 390.1367, 406.1088, С14Н25NO10, calc.,
368.1551 [M + H]+; 390.1371 [M + Na]+; 406.1110 [M + K]+.
1H NMR, : 1.21 (d, 3 H, CH3, Fuc, J = 6.5 Hz); 2.02 (s, 3 H,
NAc); 4.11 (m, 1 H, H(5), Fuc); 4.71 (d, H(1), ꢀGlcNAc,
J = 8.4 Hz); 4.92 (m, 1 H, H(1), Fuc); 5.19 (d, H(1), ꢀGlcNAc,
J = 3.4 Hz).
2ꢀAcetamidoꢀNꢀ(Nꢀbenzyloxycarbonylglycyl)ꢀ2ꢀdeoxyꢀ3ꢀOꢀ
(ꢀLꢀfucopyranosyl)ꢀꢀDꢀglucopyranosylamine (1c), []D21 –53.8
(с 1, Н2О). MS (positive ions), m/z: 558.2284, 580.2103,
596.1844, С24H35N3O12, calc. 558.2294 [M + H]+; 580.2113
[M + Na]+; 596.1852 [M + K]+. 1H NMR, : 1.18 (d, 3 H, CH3,
Fuc, J = 6.5 Hz); 2.00 (s, 3 H, NAc); 4.35 (qd, 1 H, H(5), Fuc,
J = 6.5 Hz); 5.03 (d, 1 H, H(1), Fuc, J = 3.8 Hz); 5.10—5.26 (m,
3 H, 1 H, H(1), GlcNAc, 2 H, CH2Ph); 7.35—7.48 (m, 5 H, Ph).
2ꢀAcetamidoꢀNꢀ(Nꢀbenzyloxycarbonylglycyl)ꢀ2ꢀdeoxyꢀ3,6ꢀdiꢀ
Oꢀ(ꢀLꢀfucopyranosyl)ꢀꢀDꢀglucopyranosylamine (2c), []D22–78.6
(с 1, Н2О). MS (positive ions), m/z: 726.2685, С30Н45N3O16,
calc.: 726.2692 [M + Na]+. MS (negative ions), m/z: 702.2715,
С30Н45N3O16, calc.: 702.2727 [M – H]–. 1H NMR, : 1.18 (d, 3 H,
CH3, Fuc13, J = 6.6 Hz); 1.21 (d, 3 H, CH3, Fuc16,
J = 6.6 Hz); 2.00 (s, 3 H, NAc); 4.12 (qd, 1 H, H(5), Fuc16,
J = 6.6 Hz); 4.34 (qd, 1 H, H(5), Fuc1 3, J = 6.6 Hz); 4.91
(d, 1 H, H(1), Fuc16, J = 3.6 Hz); 5.02 (d, 1 H, H(1), Fuc13,
J = 3.6 Hz); 5.09 (d, 1 H, H(1), GlcNAc, J = 10.1 Hz); 5.11,
5.17 (ABꢀsystem, 2 H, CH2Ph, J = 12.6 Hz); 7.35—7.48 (m, 5 H,
Ph). 13C NMR, : 16.41 (C(6), Fuc13); 16.54 (C(6), Fuc16);
23.31 (CH3CO); 44.98 (CH2N); 54.99 (C(2), GlcNAc); 67.98
(C(5), Fuc16); 68.18 (C(5), Fuc13); 68.58 (CH2Ph); 68.72
(C(6), GlcNAc); 69.18 (C(2), Fuc13); 69.43 (C(2), Fuc16);
69.70 (C(4), GlcNAc); 70.74 (2 C(3), 2 Fuc); 73.07 (2 C(4),
2 Fuc); 78.05 (C(5), GlcNAc); 79.66 (C(1), GlcNAc); 81.85 (C(3),
GlcNAc); 100.45 (C(1), Fuc16); 101.21 (C(1), Fuc13);
129.03; 129.68; 130.03 (6 C, C6H5); 174.21 (C=O); 175.91 (C=O).
2ꢀAcetamidoꢀNꢀ(Nꢀbenzyloxycarbonylglycyl)ꢀ2ꢀdeoxyꢀ4ꢀOꢀ
(ꢀLꢀfucopyranosyl)ꢀ3ꢀOꢀ(ꢀDꢀgalactopyranosyl)ꢀꢀDꢀglucoꢀ
pyranosylamine (3c), []D21 –40.6 (с 1, Н2О). MS (positive ions),
m/z: 720.2822, 742.2634, 758.2351, С30Н45N3O17, calc. 720.2822
[M + H]+; 742.2641 [M + Na]+; 758.2381 [M + K]+. MS (negꢀ
ative ions), m/z: 718.2675, С30Н45N3O17, calc. m/z: 718.2676
ꢀGlycopyranosylamines 1b—3b. Oligosaccharide 1a, 2a or
3a (0.1 mmol) was dissolved in 1 mL of H2O, boric acid was
added (93 mg, 1.5 mmol) and powdered ammonium carbamate
(468 mg, 6 mmol), stirred (~15 min) and kept for 4 days at ~20 С.
The residue was filtered off, washed with MeOH and the soluꢀ
tion was concentrated to ~0.3 mL at ~40 Torr. MeOH (7mL)
was added to the residue and the solution was concentrated ~0.3 mL
at ~40 Torr. This procedure was repeated several times until
total removal of ammonium carbamate, which was detected with
a pH paper placed over the solution of the compound in methaꢀ
nol, andfinally concentrated to dryness at ~10 Torr. The resiꢀ
dues were dried and the obtained ꢀglycopyranosylamines 1b—3b
were Nꢀacylated without further purification or storage.
ꢀGlycopyranosylamine 4b. Oligosaccharide 4a (36.7 mg,
0.1 mmol) was dissolved in 0.12 mL of H2O, the 0.88 mL of
MeOH and powdered ammonium carbamate (188 mg, 2.4 mmol)
were added, the reaction mixture was stirred for ~15 min and
kept for 4 days at ~20 С. The reaction mixture was diluted with
7 mL of MeOH, the solution was concentrated and the excess of
ammonium carbamate was removed with MeOH as described
earlier. The residue was dried to obtain ꢀglycopyranosylamine
4b (36 mg) contaminated with (up to 10%) of starting oligosacꢀ
charide 4a (as estimated by the visual analysis of the intensity of
spots on the electropherogram).
Nꢀ(NꢀZꢀGlycyl)ꢀꢀglycopyranosylamines 1c—4c. ꢀGlycopyꢀ
ranosylamines 1b—4b, obtained from 0.1 mmol of oligosacchaꢀ
rides 1a—4a, without further purification were dissovled in
0.15 mL of H2O, cooled with ice, 0.85 mL of DMF and 112 mg
(0.3 mmol) of NꢀZꢀglycine Nꢀhydroxysuccinimide ester were
added and stirred for 3 h at ~0 С. The reaction mixture was
diluted with 10 mL of MeOH, concentrated to ~1 mL, and this
procedure was repeated 2 times. To the stirred residue (~0.8 mL)
20 mL of Et2O was added. After the liquid became clear, it was
decanted from the oily residue. The residue was several times
washed with Et2O (5 mL), and the mixture Et2O—acetone (1 : 1),
and dried. The residue was dissolved in 1 mL of H2O and subꢀ
jected to column chromatography on С18ꢀreversed phase silica
gel in H2O in proportion 10 mg of the compound to 1 g (~3 mL)
of the phase. The elution was proceeded until the absence of
UVꢀabsorption, and then the product was eluted with 25% aqueous
MeOH. The water—methanol fractions, which contained comꢀ
pounds 1c—4c, were combined and concentrated until dryness.
The residue was dissolved in 3 mL of 10% Et3N in 50% aqueous
MeOH and kept for 3 h at ~20 С. The reaction mixture was
diluted with 10 mL of MeOH, concentrated to dryness, and this
procedure was repeated twice. The residue was dissolved in H2O
1
[M – H]–. H NMR, : 1.18 (d, 3 H, CH3, Fuc, J = 6.5 Hz);
2.00 (s, 3 H, NAc); 4.50 (d, 1 H, H(1), Gal, J = 7.6 Hz); 4.88
(qd, 1 H, H(5), Fuc, J = 6.5 Hz); 5.04 (d, 1 H, H(1), Fuc,
J = 3.6 Hz); 5.11 m, 1 H, 1(H), GlcNAc); 5.12, 5.17 (ABꢀsystem,
2 H, CH2Ph, J = 12.5 Hz); 7.33—7.49 (m, 5 H, Ph).
2ꢀAcetamidoꢀNꢀ(Nꢀbenzyloxycarbonylglycyl)ꢀ2ꢀdeoxyꢀ6ꢀOꢀ
(ꢀLꢀfucopyranosyl)ꢀꢀDꢀglucopyranosylamine (4c), []D21 –28.3
(с 1, Н2О). MS (positive ions), m/z: 558.2302, 580.2116, 596.1853,
С24Н35N3O12, calc. 558.2294 [M + H]+; 580.2113 [M + Na]+;
596.1852 [M + K]+. MS (negative ions), m/z: 556.2160.
С24Н35N3O12, calc. 556.2148 [M – H]–. 1H NMR, : 1.19 (d, 3 H,
CH3, Fuc, J = 6.3 Hz); 2.00 (s, 3 H, NAc); 4.10 (m, 1 H, H(5),
Fuc); 4.89 (d, 1 H, H(1), Fuc, J = 3.5 Hz); 5.06 (d, 1 H, H(1),
GlcNAc, J = 9.8 Hz); 5.09, 5.15 (ABꢀсsystem, 2 H, CH2Ph,
J = 12.6 Hz); 7.30—7.48 (m, 5 H, Ph).
NꢀGlycylꢀꢀglycopyranosylamines 1d—4d. Compounds 1c,
3c, 4c (0.03 mmol) and compound 2с (0.01 mmol) were disꢀ
solved in 2.5 mL (0.8 mL for 2с) of 25% aqueous MeOH, then
10% Pd/C (half the weight of compounds 1c—4c) under Ar and

1140
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 5, May, 2015
Likhosherstov et al.
Table 1. 1H NMR data (characteristic signals) for compound 1d—4d
Residue
GlcNAc
Proton
 (J/Гц)
1d
2d
3d
4d
H(1)
NAc
H(1)
CH2N
H(1)
H(5)
CH3
H(1)
H(5)
CH3
H(1)
H(5)
CH3
5.13 (J = 9.8)
5.13 (J = 10.0)
2.00
5.13 (J = 9.6)
5.09 (J = 9.7)
2.00
—
3.41
2.00
2.00
—
3.43
—
—
—
Gal
Gly
Fuc(13)
—
3.41
4.50 (J = 7.6)
3.41
—
—
—
—
—
—
5.03 (J = 3.8)
4.37 (J = 6.5)
1.18 (J = 6.5)
5.01 (J = 3.8)
4.33 (J = 6.6)
1.17 (J = 6.6)
4.91 (J = 3.8)
4.12 (J = 6.6)
1.22 (J = 6.6)
—
Fuc(16)
Fuc(14)
—
—
—
—
—
—
4.90 (J = 3.5)
4.11 (J = 6.3)
1.20 (J = 6.3)
5.04 (J = 3.6)
4.88
—
—
—
—
—
1.18 (J = 6.5)
the hydration was performed upon stirring under a weak flow of
H2 for 7 h at ~20 С. The reaction mixture was diluted with 25%
aqueous MeOH, the catalyst was removed by centrifugation,
washed with 25% aqueous MeOH and the solution was concenꢀ
trated to dryness. The residue was dissolved in 1 mL of H2O and
applied onto the C18ꢀreversed phase silica gel column (0.6×3 cm).
The product was eluted with 20 mL of H2O, the solution was
concentrated and dried. Oligosaccharides 1d—4d were obtained
with yield 95% as amorphous powder.
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ3ꢀOꢀ(ꢀLꢀfucopyranosyl)ꢀNꢀglycylꢀꢀ
Dꢀglucopyranosylamine (1d), []D22 –75.1 (с 1, Н2О). MS (posiꢀ
tive ions), m/z: 424.1907, 446.1731, 462.1471, С16Н29N3O10,
calc. m/z: 424.1926 [M + H]+; 446.1745 [M + Na]+; 462.1485
[M + K]+. MS (negative ions), m/z: 422.1793, С16Н29N3O10,
calc. 422.1780 [M – H]–.
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ3,6ꢀdiꢀOꢀ(ꢀLꢀfucopyranosyl)ꢀNꢀglyꢀ
cylꢀꢀDꢀglucopyranosylamine (2d), []D21 –107.0 (с 1, Н2О). MS
(positive ions), m/z: 570.2508, 592.2328, 608.2062, С22Н39N3O14,
calc. 570.2505 [M + H]+; 592.2324 [M + Na]+; 608.2064
[M + K]+. MS (negative ions), m/z: 568.2345, C22Н39N3O14,
calc. 568.2359 [M – H]–. 13C NMR, : 16.41 (C(6), Fuc13);
16.54 (C(6), Fuc16); 23.28 (CH3CO); 44.39 (CH2N); 55.14
(C(2), GlcNAc); 67.99 (C(5), Fuc16); 68.19 (C(5), Fuc13);
68.67 (C(6), GlcNAc); 69.17 (C(2), Fuc13); 69.42 (C(2),
Fuc16); 69.68 (C(4), GlcNAc); 70.69 (2 C(3), 2 Fuc); 73.07
(2 C(4), 2 Fuc); 78.06 (C(5), GlcNAc); 79.50 (C(1), GlcNAc);
81.92 (C(3), GlcNAc); 100.39 (C(1), Fuc16); 101.24 (C(1),
Fuc13); 176.00 (C=O); 176.18 (C=O).
References
1. M. Flowers, Adv. Carbohydr. Chem. Biochem., 1981,
39, 279.aaa
2. F. Barresi, O. Hindsgaul, J. Carohydr. Chem., 1995,
14, 1043.aaa
3. (a) H. Herzner, H. Kunz, Carbohydr. Res., 2007, 342, 541;
(b) H. Paulsen, S. Peters, T. Bielfeldt, in Glycoproteins, Eds
J. Montreuil, J. F. G. Vliegenthart, H. Schachter, Elsevier,
Amsterdam, 1995, 87.
4. L. M. Likhosherstov, O. S. Novikova, I. A. Yamskov, V. E.
Piskarev, Russ. Chem. Bull. (Int. Ed.), 2012, 61, 1816 [Izv.
Akad. Nauk, Ser. Khim., 2012, 1800].
5. L. M. Likhosherstov, O. S. Novikova, N. G. Kolotyrkina,
I. A. Yamskov, V. E. Piskarev, Russ. Chem. Bull. (Int. Ed.),
2014, 63, 507 [Izv. Akad. Nauk, Ser. Khim., 2014, 507].
6. J. Shang, V. E, Piskarev, M. Xia, P. Huang, X. Jiang, L. M.
Likhosherstov, O. S. Novikova, D. S. Newburg, D. M. Ratꢀ
ner, Glycobiology, 2013, 23, 1491.
7. S. M. Ahmed, A. J. Hall, A. E. Robinson, L. Verhoef,
P. Premkumar, U. D. Parashar, M. Koopmans, B. A. Lopman,
Lancet Infect Dis., 2014, 14, 725.
8. A. Kobata, Eur. J. Biochem., 1992, 209, 483.
9. E. Staudacher, F. Altmann, I. B. H. Wilson, L. Marz, Bioꢀ
chim. Biophs. Acta, 1999, 1473, 216.
10. K. Paschinger, D. Rendic, I. B. H. Wilson, Glycoconjugate
J., 2009, 26, 385.
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ4ꢀОꢀ(ꢀLꢀfucopyranosyl)ꢀ3ꢀOꢀ(ꢀDꢀ
galactopyranosyl)ꢀNꢀglycylꢀꢀDꢀglucopyranosylamine (3d),
[]D –62.5 (с 1, Н2О). MS (positive ions), m/z: 586.2459,
11. R. L. Shields, J. Lai, R. Keck, L. Y. O´Connell, K. Hong,
Y. G. Meng, S. H. A. Weikert, L. G. Presta, J. Biol. Chem.,
2002, 277, 26733.
23
608.2278, 624.1996, С22Н39N3О15, calc. m/z: 586.2454 [M + H]+;
608.2273 [M + Na]+; 624.2013 [M + K]+. MS (negative ions),
m/z: 584.2312, С22Н39N3О15, calc. m/z: 584.2308 [M – H]–.
2ꢀAcetamidoꢀ2ꢀdeoxyꢀ6ꢀOꢀ(ꢀLꢀfucopyranosyl)ꢀNꢀglycylꢀꢀ
Dꢀglucopyranosylamine (4d), []D20 –58.0 (с 1, Н2О). MS (posiꢀ
tive ions), m/z: 424.1926, 446.1745, 462.1481, С16Н29N3O10,
calc. 424.1926 [M + H]+; 446.1745 [M + Na]+; 462.1485 [M + K]+.
MS (negative ions), m/z: 422.1787, С16Н29N3O10, calc. 422.1780
[M – H]–.
12. K. Mori, S. Iida, N. YamaneꢀOhnuki, Y. Kanda, R. Kuniꢀ
Kamochi, R. Nakano, H. ImaiꢀNishiya, A. Okazaki,
T. Shinkawa, A. Natsume, R. Niwa, K. Shitara, M. Satoh.
Cytotechnology, 2007, 55, 109.
13. F. Altmann, Allergy Immunol., 2007, 142, 99.
14. (a) L. M. Likhosherstov, O. S. Novikova, V. N. Shibaev,
Dokl. Chem. (Engl. Transl.), 2002, 383, 89 [Dokl. Akad. Nauk,
2002, 383, 500]; (b) L. M. Likhosherstov, O. S. Novikova,
V. N. Shibaev, Dokl. Chem. (Engl. Transl.), 2003, 389, 73
[Dokl. Akad. Nauk, 2003, 389, 482].
1H NMR data for compounds 1d—4d is given in Table 1.

NꢀGlycylꢀglycosylamines of fucosaccharides
Russ.Chem.Bull., Int.Ed., Vol. 64, No. 5, May, 2015
1141
15. (a) M. DejterꢀJuszynski, H. M. Flowers, Carbohydr. Res.,
1973, 30, 287; (b) K. L. Matta, E. A. Z. Johnson, J. J. Barꢀ
low, Carbohydr. Res., 1974, 32, 396.
Kochetkov, Bull. Acad. Sci. USSR, Div. Chem. Sci.
(Engl. Transl.), 1986, 35, 1512 [Izv. Akad. Nauk, Ser. Khim.,
1986, 1663].
16. (a) J. C. Jacquinet, P. Sinay, J. Chem. Soc., Perkin Trans.1,
1979, 319; (b) R. U. Lemieux, H. Driguez, J. Am. Chem.
Soc., 1975, 97, 4063; (c) S. Rana, K. L. Matta, Carbohydr.
Res., 1983, 117, 101.
17. (a) M. DejterꢀJuszynski, H. M. Flowers, Carbohydr. Res.,
1972, 23, 41; (b) M. DejterꢀJuszynski, H. M. Flowers, Carboꢀ
hydr. Res., 1971, 18, 219; (c) S. Rana, J. J. Barlow, K. L.
Matta, Carbohydr. Res., 1982, 101, 245.
24. R. J. Ferrier, Adv. Carbohyr. Chem. Biochem., 1978, 35, 31.
25. K. Matsumura, K. Higashida, H. Ishida, Y. Hata, K. Yamaꢀ
moto, M. Shigeta, Y. MizunoꢀHorikawa, X. Wang, E. Miꢀ
yoshi, J. Gu, N.Taniguchi, J. Biol. Chem., 2007, 282, 15700.
26. I. van Die, R. D. Cummings, Glycobiology, 2010, 20, 2.
27. S. E. Newton, E. N. Meeusen, Parasite Immunol., 2003,
25, 283.
28. S. M. Haslam, G. C. Coles, E. U. Munn, T. S. Smith, H. F.
Smith, H. R. Morris, A. Dell, J. Biol. Chem., 1996, 271,
30561.
18. C. Steindl, P. Kosma, L. Marz, A. Neszmelyi, Carbohydr.
Res., 1993, 246, 353.
19. (a) M. Collot, I. B. H. Wilson, M. Bublin, K. Hoffmanꢀ
Sommergruber, J.ꢀM. Mallet, Bioorg. Med. Chem., 2011, 19,
1306; (b) J. Nakano, A. Ishiwata, H. Ohta, Y. Ito, Carbohydr.
Res., 2007, 342, 675.
20. S. Sato, M. Mori, Y. Ito, T. A. Ogawa, Carbohydr. Res.,
1973, 155, C6.
29. P. A. Belyakov, V. I. Kadentsev, A. O. Chizhov, N. G. Koloꢀ
tyrkina, A. S. Shashkov, V. P. Ananikov, Mendeleev Comꢀ
mun., 2010, 20, 125.
30. A. I. Usov, M. A. Rekhter, J. Gen. Chem. USSR (Engl. Transl.),
1969, 39 [Zh. Obshch. Khimii, 1969, 39, 912].
31. H. Lonn, Carbohydr. Res., 1985, 139, 105.
32. R. Kuhn, H. H. Baer, A. Seeliger, Ann. Chimie, 1958,
611, 236.
21. R. L. Whistler, J. N. BeMiller, Adv. Carbohyr. Chem. Bioꢀ
chem., 1958, 13, 300.
22. V. A. Derevitskaya, L. M. Likhosherstov, V. A. Schennikov,
N. K. Kochetkov, Carbohydr. Res., 1971, 20, 285.
23. (a) L. M. Likhosherstov, O. S. Novikova, V. A. Derevitskaya,
N. K. Kochetkov, Carbohydr. Res., 1986, 146, C1; (b) L. M.
Likhosherstov, O. S. Novikova, V. A. Derevitskaya, N. K.
Received August 28, 2014;
in revised form November 20, 2014