Synthesis of β-d-galactopyranosylamine derivatives with the terminal amino group in the spacer as mono-, di-, and trivalent ligands of galectins

Article abstract of DOI:10.1007/s11172-008-0345-8

Glycoclusters were obtained by N-alkylation of N-glycyl-β-d- galactopyranosylamine with N-chloroacetyl derivatives of β-d- galactopyranosylamine and N,N'-iminodiacetyl-di-β-d-galactopyranosylamine. The glycoclusters with two and three galactopyranosylamin

Full text of DOI:10.1007/s11172-008-0345-8

Russian Chemical Bulletin, International Edition, Vol. 57, No. 11, pp. 2418—2422, November, 2008
2418
Synthesis of βꢀDꢀgalactopyranosylamine derivatives
with the terminal amino group in the spacer
as monoꢀ, diꢀ, and trivalent ligands of galectins
L. M. Likhosherstov,a O. S. Novikova,a and G. V. Mokrovb
aN. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences,
47 Leninsky prosp., 119991 Moscow, Russian Federation.
Fax: +7 (499) 135 5328. Eꢀmail: knirel@ioc.ac.ru
bHigher Chemical College of the Russian Academy of Sciences,
9 Miusskaya pl., 125047 Moscow, Russian Federation
Glycoclusters were obtained by Nꢀalkylation of NꢀglycylꢀβꢀDꢀgalactopyranosylamine
with Nꢀchloroacetyl derivatives of βꢀDꢀgalactopyranosylamine and N,N´´ꢀiminodiacetylꢀ
diꢀβꢀDꢀgalactopyranosylamine. The glycoclusters with two and three galactopyranosylamine
residues and the monovalent ligand NꢀdiglycylꢀβꢀDꢀgalactopyranosylamine with an amino
group in the spacer are suitable for subsequent conjugation with carboxylꢀcontaining
physiologically active compounds.
Key words: glycoclusters, polyvalent ligands, NꢀglycylꢀβꢀDꢀgalactopyranosylamine,
NꢀchloroacetylꢀβꢀDꢀgalactopyranosylamine.
monovalent ligand NꢀdiglycylꢀβꢀDꢀgalactopyranosylꢀ
amine (4) with a spacer consisting of seven atoms with a
terminal amino group (Scheme 1).
Some tumor cells are known to have an increased
surface concentration of galectins (the proteins capable of
binding βꢀDꢀgalactoseꢀcontaining glycoconjugates).1 This
feature of tumor cells was successfully used for targeted
delivery of glycoconjugates to breast carcinoma cells.2
One of accessible ligands of galectins is the disaccharide
lactose with the terminal Oꢀlinked βꢀDꢀgalactose residue.
Earlier, we developed methods for the synthesis of monoꢀ,
diꢀ, and trivalent ligands of galectins, lactosylamine
derivatives,3,4 and obtained some monovalent glycoconꢀ
jugates of polyhedral boron compounds (with orthoꢀ
carboranylacetic acid as an example),5 which are potenꢀ
tial agents for boron neutron capture therapy of cancer.
The goal of the present work was to obtain similar
ligands of galectins from βꢀgalactopyranosylamine and
compare the efficacies of galectin binding to glycoconꢀ
jugates containing Oꢀ and Nꢀlinked βꢀDꢀgalactose residues.
One of the starting compounds for the synthesis of the
ligands was NꢀglycylꢀβꢀDꢀgalactopyranosylamine (1) preꢀ
pared earlier6 from βꢀDꢀgalactosylamine (here we syntheꢀ
sized this compound in a new, more efficient way7). It is
thought that the spacer linking a carbohydrate with a
physiologically active compound should be long enough
in order not to hinder an interaction of the carbohydrate
with the lectin.8 For this reason, the spacer in compound
1 was lengthened by coupling with Nꢀbenzyloxycarbonylꢀ
glycine (2) in the presence of N,N´´ꢀdicyclohexylcarꢀ
bodiimide (DCC) and Nꢀhydroxysuccinimide (NHS).
Hydrogenolysis of the resulting compound 3 gave the
NꢀAlkylation of amino compound 1 with Nꢀchloroꢀ
acetylꢀβꢀDꢀgalactopyranosylamine (5)9 in aqueous MeOH
in the presence of Pri2EtN gave N,N´´ꢀiminodiacetylꢀ
diꢀβꢀDꢀgalactopyranosylamine (6). Imino compound 6
was further used to both lengthen the spacer and obtain a
trivalent ligand. The spacer was lengthened by condensaꢀ
tion of compound 6 with a twofold excess of Nꢀbenzylꢀ
oxycarbonylglycylglycine (7), DCC, and NHS in DMSO—
DMF (10 : 1) at 15 °C. Few Oꢀacyl groups in Nꢀacylated
product 8 were removed by treatment with Et3N in aqueous
MeOH. Divalent ligand 9 was obtained upon Nꢀdeprotecꢀ
tion of compound 8 by hydrogenolysis over Pd/C.
To obtain a precursor of a trivalent ligand, we carried
out Nꢀchloroacetylation of imino compound 6 with
chloroacetic anhydride (10) (twofold excess) in dry DMF.
Few Oꢀacyl groups in the Nꢀacylated product were
removed by treatment with Et3N in aqueous MeOH to
yield compound 11. NꢀAlkylation of amino compound 1
with Nꢀchloroacetyl compound 11 gave imino compound
12. The spacer in compound 12 was lengthened by a
reaction with an activated ester of acid 7, namely, the
NHS ester of Nꢀbenzyloxycarbonylglycylglycine (13).
The yield of compound 14 was higher than that achieved
in a reaction of acid 7 with DCC and NHS. Hydroꢀ
genolysis of the Nꢀprotecting group in compound 14 gave
the target trivalent ligand 15.
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 11, pp. 2369—2373, November, 2008.
1066ꢀ5285/08/5711ꢀ2418 © 2008 Springer Science+Business Media, Inc.

NꢀGlycyl(galactosylamine) glycoclusters
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 11, November, 2008 2419
The structures of the compounds obtained were conꢀ
firmed by data from elemental analysis and NMR spectra.
The H NMR spectra of all compounds with one, two,
and three βꢀgalactosylamine residues contain signals for
the H(2)—H(6) protons of the galactose residues at
δ 3.60—4.00, which were not analyzed by us any further.
1
Scheme 1
Z = PhCH2OCO
Reagents and conditions: a, DCC, NHS, DMSO; b, H2, Pd/C, H2O; c, Pri2EtN, MeOH—H2O (3 : 2); d, DMF;
e, Pri2EtN, DMSO; f, DMSO—DMF (2 : 3)

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Russ.Chem.Bull., Int.Ed., Vol. 57, No. 11, November, 2008
Likhosherstov et al.
AgNO3—KOH (see Ref. 11). Water was determined according
to the Fischer method. Column chromatography on silica
gel Sꢀ18 was monitored by UV absorption at 206 and 260 nm.
Nꢀ(NꢀBenzyloxycarbonyldiglycyl)ꢀβꢀDꢀgalactopyranosylꢀ
amine (3). A mixture of NꢀglycylꢀβꢀDꢀgalactopyranosylamine
(1) (0.35 g, 1.5 mmol), Nꢀbenzyloxycarbonylglycine (2) (0.32 g,
1.5 mmol), and NHS (0.19 g, 1.65 mmol) was dissolved in dry
DMSO (10 mL) heated to 50 °C. The resulting solution was
cooled to 15 °C and DCC (0.37 g, 1.8 mmol) was added. The
reaction mixture was stirred for 45 min and kept at 20 °C for
20 h. The precipitate of N,N´ꢀdicyclohexylurea was filtered off
and washed with DMSO (2×0.75 mL). The filtrate was added to
stirred Et2O (120 mL) and the clarified liquid was decanted
from an oily precipitate. The precipitate was repeatedly trituꢀ
rated with Et2O (25 mL each portion) into a viscous mass,
which was repeatedly treated with acetone (15 mL each porꢀ
tion). The resulting powder was filtered off, dried, and dissolved
in water (1 mL). Methanol (5×3 mL) was added with stirring
and heating to 40 °C. The solution was kept at 5 °C for 48 h. The
precipitate that formed was filtered off, washed with MeOH and
Et2O, and dried. The yield of compound 3 was 0.48 g (75%), an
amorphous solid, [α]D21 +10.2 (c 1, H2O). Found (%): C, 50.01;
H, 6.06; N, 9.84. C18H25N3O9. Calculated (%): C, 50.58;
H, 5.90; N, 9.83. 1H NMR, δ: 3.62—3.83 (m, 5 H); 3.92 (s,
2 H, CH2NCOCH2); 3.98 (br.s, 1 H, H(4)); 4.02 (s, 2 H,
CH2NHCOCH2); 4.95 (d, 1 H, H(1), J = 9.0 Hz); 5.17 (s,
2 H, CH2Ph); 7.44 (br.s, 5 H, Ph).
The spectra show some distinctions in the character and
number of the signals for the H(1) protons in compounds
with two (except for 6) and three galactosylamine resiꢀ
dues. The spectra of compounds 8, 9, and 11 each conꢀ
taining two βꢀgalactose residues exhibit two doublets with
Δδ 0.05—0.07 ppm. The spectra of compounds containing
three βꢀgalactose residues show a doublet and a doubleꢀ
intensity doublet with Δδ 0.04 ppm (12) and multiplets at
δ 4.89—5.02 (14) and 4.94—5.06 (15).
The signals for the CH2 protons of the spacer appear
in a wide range (δH 3.44—5.20). Part of these signals
(mainly broadened singlets) in the spectra of compounds
3, 4, 6, 8, 9, and 11 were assigned to particular CH2
groups (see Experimental) from both the literature3,5,6,9
and our experimental data (appearance or disappearance
of the signals as the result of the reactions). We did not
assign the signals to particular CH2 groups in compounds
14 and 15 (except for CH2Ph and CH2NH2, respectively)
because the CH2 protons produced a complex spectrum
with numerous signals. The presence of many signals for
the CH2 protons and a multiplet for the H(1) protons of
three βꢀgalactose residues in the spectra of compounds 14
and 15, as in analogous lactosylamine derivatives,4 is
attributable to the formation of rotamers because of the
presence of two disubstituted amide groups (see Ref. 10
and references therein).
The structure of trivalent ligand 15 was additionally
confirmed by its 13C NMR spectrum. The spectrum shows
signals for six carbon atoms of three βꢀDꢀgalactopyranosylꢀ
amine residues and lowꢀintensity signals for the CH2 and
CO carbon atoms: four signals (including two broadened
ones) due to six CH2 groups and six signals due to six
CO groups.
Thus, using the previously developed methods, we
obtained monoꢀ, diꢀ, and trivalent ligands of galectins,
βꢀgalactopyranosylamine derivatives, with the terminal
amino group in the spacer. These ligands are suitable for
conjugation with carboxylꢀcontaining physiologically
active compounds. The compounds obtained will allow
further investigations (see Ref. 5 and references therein)
aimed at the synthesis of new monoꢀ and polyvalent
glycoconjugates of polyhedral boron compounds, which
are of interest as potential agents for boron neutron
capture therapy of cancer.
NꢀDiglycylꢀβꢀDꢀgalactopyranosylamine (4). 10% Pd/C
(0.21 g) was added under argon to a solution of compound 3
(0.415 g, 0.97 mmol) in water (10 mL). Hydrogenation was
carried out by passing a weak flow of H2 through the vigorously
stirred mixture at 20 °C for 8 h. The reaction mixture was filꢀ
tered through silica gel Cꢀ18 (4 g). The sorbent was washed with
water (80 mL) and the solution was concentrated to a syrup,
which partially crystallized. The syrup was kept at 5 °C for 16 h.
Then MeOH (5 mL) was added to the resulting crystalline solid
and the precipitate was filtered off, washed with MeOH and
Et2O, and dried. The yield of compound 4 was 0.22 g (78%),
18
m.p. 165—166 °C, [α]D +12.2 (c 1, H2O). Found (%):
C, 40.47; H, 6.61; N, 13.84. C10H19N3O7. Calculated (%):
C, 40.95; H, 6.53; N, 14.33. 1H NMR, δ: 3.48 (s, 2 H, CH2NH2);
3.61—3.83 (m, 5 H); 3.97 (m, 1 H, H(4)); 4.03 (s, 2 H, CH2N);
4.95 (d, 1 H, H(1), J = 9.0 Hz).
N,NꢀBis[Nꢀ(βꢀDꢀgalactopyranosyl)carbamoylmethyl]amine
(6).
A mixture of compound 1 (0.82 g, 3.47 mmol)
and NꢀchloroacetylꢀβꢀDꢀgalactopyranosylamine (5) (0.89 g,
3.49 mmol) in 60% aqueous MeOH (13 mL) and Pri EtN
2
(0.66 mL, 3.88 mmol) was heated with periodical stirring in a
screwꢀcapped test tube at 70 °C for 20 h. The reaction mixture
was diluted with water (10 mL) and concentrated to 10 mL. The
residue was diluted with water (15 mL), stirred with the anion
exchange resin Dowex 1w×8 (OH–) (18 mL) for 10 min, and
again diluted with water (50 mL). The liberated Pri2EtN was
codistilled with water, while concentrating the suspension to
35 mL. The resin was filtered off and washed with water
(150 mL). The filtrate and the aqueous eluate were concentrated
to 50 mL and stirred with the cation exchange resin Dowex
50w×8 (H+) (30 mL) for 1 h. The resin was filtered off, washed
with water (300 mL) and 0.5 M aqueous pyridine (250 mL). The
latter eluate was evaporated to dryness. The residue was disꢀ
solved in water (3 mL). Then MeOH (40 mL) was added with
Experimental
1H and 13C NMR spectra were recorded in D2O at 24 °C on
a Bruker WMꢀ250 spectrometer (250 (1H) and 62.7 MHz (13C))
with acetone as the external standard. Optical rotation was meaꢀ
sured on a PUꢀ07 polarimeter (Russia). The course of the reacꢀ
tions and the isolation of products were monitored by electroꢀ
phoresis (30 V cm–1, 1 h) on Filtrak FN1 paper in pyridinium
acetate buffer (0.025 M Py, pH 4.5). The products were visualꢀ
ized with ninhydrin and the sequence of reagents KIO4—

NꢀGlycyl(galactosylamine) glycoclusters
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 11, November, 2008 2421
stirring and the solution was kept at 5 °C for 16 h. The precipiꢀ
tate that formed was filtered off, washed with MeOH and Et2O,
and dried. The yield of compound 6 was 0.95 g (59%), an amorꢀ
Nꢀ(Chloroacetyl)ꢀN,Nꢀbis[Nꢀ(βꢀDꢀgalactopyranosyl)carbaꢀ
moylmethyl]amine (11). A suspension of compound 6 (0.685 g,
1.47 mmol) in dry DMF (7 mL) was heated at 70 °C to homoꢀ
genization and cooled with ice. Chloroacetic anhydride (10)
(0.311 g, 1.8 mmol) was added and the mixture was stirred
at 0 °C for 3 h. Then another portion of anhydride 10 (0.178 g,
1.04 mmol) was added with stirring and the mixture was kept at
0 °C for 3 h. The mixture was diluted with MeOH (10 mL) and
kept at 20 °C for 2 h. Then Et2O (70 mL) was added with stirring
and the mixture was kept at 5 °C for 16 h. The gelꢀlike precipiꢀ
tate that formed was filtered off, washed with MeOH—Et2O (2 : 1),
MeOH—Et2O (1 : 1), and Et2O, and dried. The residue was
dissolved in water (3 mL), MeOH (3 mL) containing Et3N
(0.6 mL) was added, and the solution was kept at 20 °C for 3 h.
The precipitate that formed was diluted with MeOH (9 mL),
filtered off, washed with 80% aqueous MeOH, then MeOH, and
Et2O, and dried. The residue was heated in water (5 mL) to
homogenization. The resulting solution was diluted with MeOH
(25 mL), heated to homogenization, and kept at 5 °C for 16 h.
The precipitate that formed was filtered off, washed with 80%
aqueous MeOH, then MeOH, and Et2O, and dried. The yield
21
phous solid, [α]D +16.2 (c 1, H2O). Found (%): C, 41.59; H,
6.84; N, 8.61; H2O, 2.35. C16H29N3O12•0.5H2O. Calculated
(%): C, 41.38; H, 6.51; N, 9.05; H2O, 1.94. 1H NMR, δ: 3.46 (s,
4 H, 2 CH2N); 3.61—3.83 (m, 10 H); 3.98 (br.s, 2 H, 2 H(4));
4.96 (d, 2 H, 2 H(1), J = 9.0 Hz).
Nꢀ(NꢀBenzyloxycarbonyldiglycyl)ꢀN,Nꢀbis[Nꢀ(βꢀDꢀgalactoꢀ
pyranosyl)carbamoylmethyl]amine (8). A mixture of compound 6
(0.228 g, 0.5 mmol), Nꢀbenzyloxycarbonylglycylglycine (7)
(0.266 g, 1 mmol), and NHS (0.134 g, 1.16 mmol) was dissolved
in dry DMSO (1.7 mL) and DMF (0.17 mL). The solution was
cooled to 10 °C and DCC (0.27 g, 1.31 mmol) was added. The
reaction mixture was stirred for 45 min and then kept at 15 °C
for 60 h. The precipitate of N,N´ꢀdicyclohexylurea was filtered
off and washed with DMSO (2×0.5 mL). The filtrate was added
to stirred Et2O (35 mL) and the clarified liquid was decanted
from the oily precipitate. The precipitate was repeatedly trituꢀ
rated with Et2O (10 mL each portion) into a viscous mass,
which was repeatedly treated with hot acetone (10 mL each
portion). The resulting powder was filtered off, washed with
Et2O, dried, and dissolved in a mixture of water (2.5 mL) and
MeOH (2.5 mL). The solution was kept with Et3N (0.5 mL) at
20 °C for 3 h, diluted with MeOH (10 mL), and concentrated to
1 mL. The procedure was repeated twice. In the last run, the
solution was concentrated to dryness. The residue was washed
with PriOH (5×7 mL) and Et2O, dried, dissolved in water
(5 mL), and filtered through silica gel Cꢀ18 (10 g). The sorbent
was washed with water (300 mL), 10% aqueous MeOH
(220 mL), and 20% aqueous MeOH (200 mL). The water—
methanol fractions were concentrated to dryness. The residue
was dissolved with heating in MeOH (5 mL) and the solution
was kept at 5 °C for 16 h. The precipitate that formed was
filtered off, washed with MeOH and Et2O, and dried. The
yield of compound 8 was 0.2 g (56%), m.p. 164—165 °C,
[α]D21 +11.5 (c 1, H2O). Found (%): C, 45.95; H, 6.20; N, 9.40;
H2O, 3.37. C28H41N5O16•1.5H2O. Calculated (%): C, 46.03;
H, 6.07; N, 9.58; H2O, 3.70. 1H NMR, δ: 3.61—3.83 (m, 10 H);
3.92 (br.s, 2 H, CH2NCOCH2); 3.99 (br.s, 2 H, 2 H(4));
4.12—4.27 (m, 4 H, 2 CH2); 4.41 (m, 2 H, CH2); 4.96 (d, 1 H,
H(1), J = 9.0 Hz); 5.03 (d, 1 H, H(1), J = 9.0 Hz); 5.18 (s, 2 H,
CH2Ph); 7.46 (br.s, 5 H, Ph).
21
of compound 11 was 0.63 g (78%), an amorphous solid, [α]D
+15.4 (c 1, H2O). Found (%): C, 39.19; H, 6.06; Cl, 6.39;
N, 7.55; H2O, 3.47. C18H30ClN3O13•H2O. Calculated (%):
C, 39.31; H, 5.86; Cl, 6.45; N, 7.64; H2O, 3.27. 1H NMR, δ:
3.60—3.82 (m, 10 H); 3.97 (m, 2 H, 2 H(4)); 4.23, 4.32
(AB system, 2 H, CH2Cl, J = 17.2 Hz); 4.48 (br.s, 2 H, CH2);
4.50 (br.s, 2 H, CH2); 4.93 (d, 1 H, H(1), J = 9.0 Hz); 4.98 (d,
1 H, H(1), J = 9.0 Hz).
Nꢀ[Nꢀ(βꢀDꢀGalactopyranosyl)carbamoylmethyl]ꢀNꢀ{N,Nꢀ
bis[Nꢀ(βꢀDꢀgalactopyranosyl)carbamoylmethyl]carbamoylꢀ
methyl}amine (12).
A mixture of compound 1 (0.44 g,
1.86 mmol), chloroacetyl derivative 11 (0.5 g, 0.91 mmol) in dry
DMSO (5.6 mL), and Pri2EtN (0.161 mL, 0.95 mmol) was
heated in a screwꢀcapped test tube at 70 °C for 16 h. On cooling
to 20 °C, Et2O (50 mL) was added with stirring. The clarified
liquid was decanted from the oily precipitate. The precipitate
was repeatedly triturated with Et2O (15 mL each portion) into a
viscous mass, which was dissolved in water (50 mL). The soluꢀ
tion was stirred with the cation exchange resin Dowex 50w×2
(H+) (50 mL) for 1 h. The resin was filtered off and washed with
water (500 mL) and 0.5 M aqueous pyridine (500 mL). The
pyridine eluate (250 mL) containing the target product was
evaporated to dryness. The residue was dissolved in water
(3 mL) and then MeOH (30 mL) was added with stirring. The
mixture was kept at 5 °C for 16 h. The precipitate that formed
was filtered off, washed with MeOH and Et2O, and dried. The
yield of compound 12 was 0.51 g (75%), an amorphous solid,
[α]D21 +16.6 (c 1, H2O). Found (%): C, 41.65; H, 6.46; N, 9.10;
H2O, 2.13. C26H45N5O19•H2O. Calculated (%): C, 41.65; H,
Nꢀ(Diglycyl)ꢀN,Nꢀbis[Nꢀ(βꢀDꢀgalactopyranosyl)carbamoylꢀ
methyl]amine (9). 10% Pd/C (0.07 g) was added under argon
to a solution of compound 8 (0.135 g, 0.185 mmol) in water
(3.5 mL). Hydrogenation was carried out by passing a weak flow
of H2 through the vigorously stirred mixture at 20 °C for 8 h. The
reaction mixture was filtered through silica gel Cꢀ18 (2 g). The
sorbent was washed with water (70 mL) and the solution was
concentrated to 0.5 mL. Methanol (10 mL) was added dropwise
with stirring at 50 °C and the solution was kept at 5 °C for 16 h.
The precipitate that formed was filtered off, washed with MeOH
and Et2O, and dried. The yield of compound 9 was 0.09 g (83%),
1
6.32; N, 9.34; H2O, 2.40. H NMR, δ: 3.41 (s, 2 H, CH2NH);
3.58 (s, 2 H, CH2NH); 3.61—3.82 (m, 15 H); 3.97 (m, 3 H,
3 H(4)); 4.14, 4.25 (AB system, 2 H, CH2N, J = 27.5 Hz); 4.31
(br.s, 2 H, CH2N); 4.94 (d, 2 H, 2 H(1), J = 9.0 Hz); 4.98 (d,
1 H, H(1), J = 9.0 Hz).
18
an amorphous solid, [α]D +13.7 (c 1, H2O). Found (%):
C, 40.75; H, 6.73; N, 11.40; H2O, 2.83. C20H35N5O14•H2O.
Calculated (%): C, 40.88; H, 6.35; N, 11.92; H2O, 3.07.
1H NMR, δ: 3.44 (br.s, 2 H, CH2NH2); 3.61—3.83 (m, 10 H);
3.97 (br.s, 2 H, 2 H(4)); 4.18 (m, 4 H, 2 CH2); 4.38 (br.s,
2 H, CH2); 4.93 (d, 1 H, H(1), J = 9.0 Hz); 4.98 (d, 1 H, H(1),
J = 9.0 Hz).
Nꢀ(NꢀBenzyloxycarbonyldiglycyl)ꢀNꢀ[Nꢀ(βꢀDꢀgalactopyraꢀ
nosyl)carbamoylmethyl]ꢀNꢀ{N,Nꢀbis[Nꢀ(βꢀDꢀgalactopyranosyl)ꢀ
carbamoylmethyl]carbamoylmethyl}amine (14). Compound 12
(0.22 g, 0.29 mmol) was dissolved at 50 °C in dry DMSO
(1.2 mL). On cooling to 15 °C, a solution of Nꢀhydroxysuccinꢀ
imide ester of Nꢀbenzyloxycarbonylglycylglycine (13) (0.218 g,

2422
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 11, November, 2008
Likhosherstov et al.
0.6 mmol) in DMF (1.8 mL) was added. The reaction mixture
was kept at 20 °C for 60 h. Another portion of DMF (0.9 mL)
containing compound 13 (0.11 g, 0.3 mmol) was added. The
mixture was kept at 20 °C for an additional 24 h and added to
stirred Et2O (50 mL). The clarified liquid was decanted from the
oily precipitate. The precipitate was repeatedly triturated with
Et2O (10 mL each portion) into a viscous mass, which was
repeatedly treated with hot acetone (10 mL each portion). The
resulting powder was filtered off, dried, and dissolved in a mixꢀ
ture of water (5 mL) and MeOH (5 mL). The solution was
treated with Et3N (1 mL) at 20 °C for 3 h, diluted with MeOH
(15 mL), and concentrated to 2 mL. The procedure was
repeated twice. In the last run, the solution was concentrated to
dryness. The residue was dissolved in MeOH (10 mL). Then
PriOH (10 mL) was added with stirring and the mixture was kept
at 5 °C for 16 h. The precipitate that formed was filtered off,
washed with MeOH—PriOH (1 : 1) and Et2O, and dried. The
residue was dissolved in water (5 mL) and filtered through silica
gel Cꢀ18 (20 g). The sorbent was washed with water (600 mL)
and 25% aqueous MeOH (400 mL). The water—methanol fracꢀ
tions were concentrated to dryness. The yield of compound 14
This work was financially supported by the Russian
Foundation for Basic Research (Project No. 07ꢀ03ꢀ00120).
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Akad. Nauk, Ser. Khim., 2008, 647 [Russ. Chem. Bull., Int.
Ed., 2008, 57, 660].
5. L. M. Likhosherstov, O. S. Novikova, L. O. Kononov, A. V.
Orlova, I. B. Sivaev, V. I. Bregadze, Izv. Akad. Nauk, Ser.
Khim., 2007, 2033 [Russ. Chem. Bull., Int. Ed., 2007, 56,
2105].
23
was 0.22 g (73%), an amorphous solid, [α]D + 10.3 (c 0.5,
H2O). Found (%): C, 43.72; H, 6.28; N, 9.34; H2O, 5.68.
C38H57N7O23•3H2O. Calculated (%): C, 44.14; H, 6.14;
N, 9.48; H2O, 5.23. 1H NMR, δ: 3.60—3.83 (m, 15 H); 3.92 (s,
2 H, CH2) 3.98 (m, 3 H, 3 H(4)); 4.04—4.47 (m, 9 H); 4.56
(br.s, 1 H); 4.89—5.02 (m, 3 H, 3 H(1)); 5.18 (s, 2 H, CH2Ph);
7.45 (br.s, 5 H, Ph).
6. L. M. Likhosherstov, O. S. Novikova, A. O. Zheltova, V. N.
Shibaev, Izv. Akad. Nauk, Ser. Khim., 2000, 1461 [Russ.
Chem. Bull., 2000, 49, 1454 (Engl. Transl.)].
7. L. M. Likhosherstov, O. S. Novikova, V. N. Shibaev, Dokl.
Akad. Nauk, 2002, 383, 500 [Dokl. Chem., 2002, 383, 89
(Engl. Transl.)].
Nꢀ(Diglycyl)ꢀNꢀ[Nꢀ(βꢀDꢀgalactopyranosyl)carbamoylꢀ
methyl]ꢀNꢀ{N,Nꢀbis[Nꢀ(βꢀDꢀgalactopyranosyl)carbamoylꢀ
methyl]carbamoylmethyl}amine (15) was obtained from comꢀ
pound 14 (0.188 g, 0.182 mmol) by hydrogenolysis in the presꢀ
ence of 10% Pd/C (0.11 g) as described for compound 9. The
yield of compound 15 was 0.14 g (91%), an amorphous solid,
8. G. Magnusson, A. Ya. Chernyak, J. Kihlberg, L. O. Kononov,
in Neoglycoconjugates: Preparation and Application, Eds
Y. C. Lee, R. T. Lee, Academic Press Inc., San Diego,
California, 1994, p. 53.
9. L. M. Likhosherstov, O. S. Novikova, V. N. Shibaev, N. K.
Kochetkov, Izv. Akad. Nauk, Ser. Khim., 1996, 1848 [Russ.
Chem. Bull., 1996, 45, 1760 (Engl. Transl.)].
15
[α]D + 12.7 (c 1, H2O). Found (%): C, 42.53; H, 6.19; N,
11.46. C30H51N7O21. Calculated (%): C, 42.60; H, 6.08;
N, 11.59. 1H NMR, δ: 3.53 (br.s, 2 H, CH2NH2); 3.63—3.84
(m, 15 H); 3.99 (m, 3 H, 3 H(4)); 4.10—4.46 (m, 7 H); 4.58
(br.s, 1 H); 4.94—5.06 (m, 3 H, 3 H(1)). 13C NMR, δ: 42.3
(CH2NH2); 44.9 (CH2NH); 52.9 (br.s, CH2NCH2); 53.2 (br.s,
CH2NCH2); 62.5 (C(6)); 70.2 (C(4)): 70.9 (C(2)); 74.9
(C(3)); 78.3 (C(5)); 81.3 (C(1)); 172.6; 172.8; 173.2; 173.4;
173.5; 173.7 (6 CO).
10. U. K. Saha, R. Roy, Tetrahedron Lett., 1997, 38, 7697.
11. A. I. Usov, M. A. Rekhter, Zh. Obshch. Khim., 1969, 39, 912
[J. Gen. Chem. USSR, 1969, 39 (Engl. Transl.)].
Received May 22, 2008