New effective synthesis of (N-acetyl- and N-stearoyl-2-amino-2-deoxy-β-D-glucopyranosyl)-(1→4)N-acetyln or-muramoyl-L-2-aminobutanoyl-D-isoglutamine, analogs of GMDP with immunopotentiating activity

Article abstract of DOI:10.1135/cccc19980577

Ethyl 3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (5), prepared by benzylation of ethyl 2-deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (4), was transformed by reaction with bromine into 3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-β-

Full text of DOI:10.1135/cccc19980577

Synthesis of Analogs of GMDP
577
NEW EFFECTIVE SYNTHESIS OF (N-ACETYL- AND N-STEAROYL-
2- AMINO-2-DEOXY-β-D-GLUCOPYRANOSYL)-(1→4)-N-ACETYLNOR-
MURAMOYL-L-2-AMINOBUTANOYL-D-ISOGLUTAMINE, ANALOGS
OF GMDP WITH IMMUNOPOTENTIATING ACTIVITY
a1
b1
a2
a3
Miroslav LEDVINA , Daniel ZYKA , Jan JEZEK , Tomas TRNKAb2 and David SAMAN
a Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
166 10, Prague 6, Czech Republic; e-mail: 1 ledvina@uochb.cas.cz, 2 jezek@uochb.cas.cz,
3 saman@uochb.cas.cz
b Department of Organic Chemistry, Charles University, 128 40 Prague 2, Czech Republic;
e-mail: 1 zyka@mail.natur.cuni.cz, 2 trnka@mail.natur.cuni.cz
Received January 15, 1998
Accepted March 14, 1998
Ethyl 3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (5), prepared by benzyla-
tion of ethyl 2-deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (4), was transformed by reaction
with bromine into 3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-β-D-glucopyranosyl bromide (6). Thio-
glycoside 5 in the presence of methyl triflate and glycosylbromide 6 in the presence of silver triflate
were used as glycosyl donors for condensation with benzyl 2-acetamido-3-O-allyl-6-O-benzyl-2-
deoxy-α-D-glucopyranoside (7), to give benzyl 2-acetamido-3-O-allyl-6-O-benzyl-4-O-(3,4,6-tri-O-
benzyl-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-2-deoxy-α-D-glucopyranoside (8). Its reductive
dephthaloylation with NaBH4/AcOH afforded benzyl 2-acetamido-3-O-allyl-4-O-(2-amino-3,4,6-tri-
O-benzyl-2-deoxy-β-D-glucopyranosyl)-6-O-benzyl-2-deoxy-α-D-glucopyranoside (11). Compound 11
was N-acylated to give benzyl 2-acetamido-4-O-(2-acylamino-3,4,6-tri-O-benzyl-2-deoxy-β-D-gluco-
pyranosyl)-3-O-allyl-6-O-benzyl-2-deoxy-α-D-glucopyranosides (12a) or (12b). These compounds
were converted into corresponding benzyl 2-acetamido-4-O-(2-acylamino-3,4,6-tri-O-benzyl-2-deoxy-
β-D-glucopyranosyl)-6-O-benzyl-3-O-carboxymethyl-2-deoxy-α-D-glucopyranosides which, by con-
densation with H-L-Abu-D-isoGln(OBzl) followed by hydrogenolysis of protective benzyl groups,
furnished glycopeptides 16a and 16b. Intramolecular O→N migration of the allyl protecting group
followed by its reduction to the propyl residue by reaction of compound 8 with hydrazine or hydra-
zinium acetate, to give benzyl 2-acetamido-4-O-(3,4,6-tri-O-benzyl-2-deoxy-2-propylamino-β-D-glu-
copyranosyl)-6-O-benzyl–2-deoxy-α-D-glucopyranoside (9), is also described.
Key words: Carbohydrates; Glycosides; Aminosugars; Oligosaccharides; Muramyl glycopeptides; Immuno-
stimulators.
Some time ago we described the synthesis of (2-acetamido-2-deoxy-β-D-glucopyrano-
syl)-(1→4)-N-acetylnormuramoyl-L-2-aminobutanoyl-D-isoglutamine (16a, β-D-GlcNAc-
(1→4)-norMurNAc-L-Abu-D-isoGln), an analogue of GMDP (β-D-GlcNAc-(1→4)-
MurNAc-L-Ala-D-isoGln) modified both in the sugar and peptide parts of the molecule,
and its lipophilic derivative (2-deoxy-2-stearamido-β-D-glucopyranosyl)-(1→4)-N-ace-
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

578
Ledvina, Zyka, Jezek, Trnka, Saman:
tylnormuramoyl-L-2-aminobutanoyl-D-isoglutamine* (16b, β-D-GlcNstearoyl-(1→4)-
norMurNAc-L-Abu-D-isoGln), bearing a bulky stearoyl residue on the NH2 group of
glucosamine subunit (refs1–3). Both compounds, compared with MDP (muramoyl-di-
peptide) and GMDP, exhibit higher immunopotentiating activity and their undesirable
side effects are suppressed (e.g., they are not pyrogenic). Immunoadjuvant activity of
the lipophilic derivative 16b is fully comparable with the activity of FCA (Freund’s
Complet Adjuvant). Compound 16a and especially its lipophilic derivative 16b stimu-
1
2
3
1
OBzl
O
1, R = Ac, R = R = H, X = OAc
OR
1
2
3
X
2, R = Ac, R + R = Pht, X = OAc
O
1
O
1
2
3
OR
3, R = Ac, R + R = Pht, X = SEt
OBzl
NHAc
CH CH=CH
HO
1
2
3
1
4, R = H, R + R = Pht, X = SEt
R O
2
3
NR R
1
2
3
5, R = Bzl, R + R = Pht, X = SEt
2
2
1
2
3
7
6, R = Bzl, R + R = Pht, X = Br
OBzl
OBzl
O
OBzl
O
1
OR
O
OBzl
NHAc
BzlO
3
2
3
NR R
1
2
3
1
2
13a, R = R = H, R = Ac
8, R = All, R = R = Pht
1
2
3
1
2
3
13b, R = R = H, R = stearoyl
9, R = H, R = H, R = (CH ) CH
2 2
3
1
2
3
1
2
3
14a, R = CH COOH, R = H, R = Ac
2
10, R = R = R = H
1
2
3
1
2
3
14b, R = CH COOH, R = H, R = stearoyl
2
11, R = All, R = R = H
1
2
3
1
2
3
15a, R = CH CO-L-Abu-D-isoGln(OBzl), R = H, R = Ac
12a, R = All, R = H, R = Ac
2
1
2
3
1
2
3
12b, R = All, R = H, R = stearoyl
15b, R = CH CO-L-Abu-D-isoGln(OBzl), R = H, R = stearoyl
2
OH
O
OH
O
OH
O
OH
O
HO
NHR
NHAc
CH CO-L-Abu-D-isoGln
All = CH CH=CH
2
2
2
Bzl = CH C H
6 5
16a, R = Ac
16b, R = stearoyl
2
Pht = phthaloyl
* Normuramic acid is the trivial name for 2-amino-3-O-carboxymethyl-2-deoxy-D-glucopyranose. The
symbols and abbreviations obey the published recommendations (Biochemical Nomenclature and Related
Documents. International Union of Biochemistry, London 1978).
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

Synthesis of Analogs of GMDP
579
late haemopoiesis and possess significant protective and therapeutic effects in radiation
injury and restore lymphopenia caused by some xenobiotics (for review, see refs4,5).
Now we describe a new effective preparation of disaccharide precursors 12a and 12b
from which compounds 16a and 16b can be prepared. For the synthesis of the key
disaccharide 8 we chose an approach based on condensation of monosaccharide units
with protecting groups representing an orthogonal system. As glycosyl acceptor we
used benzyl 2-acetamido-3-O-allyl-6-O-benzyl-2-deoxy-α-D-glucopyranoside (7, ref.1)
and as a glycosyl donor, or its precursor, we used ethyl 3,4,6-tri-O-benzyl-2-deoxy-2-
phthalimido-1-thio-β-D-glucopyranoside (5). The use of glycosyl donor protected by
O-benzyl groups instead of the often used O-acetyl groups enabled us to avoid the
cumbersome exchange of the protecting groups after the demanding glycosylation step
(see, e.g., refs1,6). The glycosyl donor 5 was obtained by O-benzylation of ethyl 2-
deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (4, ref.7) by benzylbromide in N,N-
dimethylformamide in the presence of sodium hydride.
Ethyl thioglycoside 4 was prepared from 1,3,4,6-tetra-O-acetyl-2-amino-2-deoxy-β-
D-glucopyranose (1) in three steps by a modified procedure7,8. 1,3,4,6-Tetra-O-acetyl-
2-deoxy-2-phthalimido-β-D-glucopyranose (2) was obtained by the reaction of amine 1
with phthalic anhydride in dichloromethane in the presence of 4-dimethylaminopy-
ridine (DMAP) and subsequent cyclization of the intermediate phthalamic acid with
acetic anhydride. Ethyl chloroformate and triethylamine used in the cyclization step in
ref.7, were replaced by acetic anhydride in the presence of DMAP with the aim to
eliminate the undesirable reaction leading to isophthalimide9,10. Our procedure, in com-
parison with described9,11,12 ones, gave phthalimido derivative 2 in consistently high
yield (82%), and is also suitable for its preparation in a larger scale. The reaction of the
phthalimido derivative 2 with ethanethiol in dichloromethane promoted by titanium
tetrachloride afforded thioglycoside 3. The O-acetyl groups of 3 were removed with
sodium methoxide in methanol to give compound 4.
Methyl triflate-promoted glycosylation13,14 of glycosyl acceptor 7 with glycosyl
donor 5 afforded benzyl 2-acetamido-3-O-allyl-6-O-benzyl-4-O-(3,4,6-tri-O-benzyl-2-
deoxy-2-phthalimido-β-D-glucopyranosyl)-2-deoxy-α-D-glucopyranoside (8) with the
yield of only 20%. Owing to the low effectivity of the glycosylation proceeding via
sulfonium ion, we used an alternative approach. Thioglycoside 5 was transformed with
bromine in dichloromethane at 0 °C (ref.15) to 3,4,6-tri-O-benzyl-2-deoxy-2-phthal-
imido-β-D-glucopyranosylbromide (6), which by reaction with glycosyl acceptor 7 in
the presence of silver triflate (molar ratio 3 : 2 : 3) in dichloromethane at –45 °C
afforded the desired disaccharide 8 in satisfactory yield of 73%. This reaction was
carried out without base because, in the case of little reactive acceptor1,2, the base acts
as a glycosylation inhibitor.
The attempt to dephthaloylate disaccharide 8 with butylamine in boiling methanol,
i.e., under the conditions described1 for the cleavage of the phthalimido group from
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

580
Ledvina, Zyka, Jezek, Trnka, Saman:
benzyl 2-acetamido-4-O-(3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-β-D-glucopyrano-
syl)-3-O-allyl-6-O-benzyl-2-deoxy-α-D-glucopyranoside, was not successful. This ne-
gative result is probably due to the presence of alkali-stable benzyl group on the vicinal
3′-OH group in disaccharide 8. In the case of the above mentioned disaccharide, which
is protected by alkali-labile O-acetyl groups, the acetyls are preferentially cleaved by
butylamine and the free 3′-OH group can participate in the dephthaloylation step. The
strong accelerating effect of vicinal 3′-OH group on base-catalyzed N-deacetylation of
2-acetamido-2-deoxy-D-glucopyranosyl residue has been described3. Also the attempt
to dephthaloylate compound 8 under harsher conditions, i.e. by hydrazine in boiling
ethanol, was not successful. Under these conditions, the dephthaloylation was followed
by the O→N migration and reduction of the allyl protecting group to the propyl
residue, and as a product, the benzyl 2-acetamido-4-O-(3,4,6-tri-O-benzyl-2-deoxy-2-
propylamino-β-D-glucopyranosyl)-6-O-benzyl-2-deoxy-α-D-glucopyranoside (9) was
only isolated. An analogous result was obtained also by dephthaloylation of compound
8 with hydrazine acetate in boiling ethanol. In the literature, only migration of the allyl
group to oxygen, nitrogen, carbon, sulfur and hydride nucleophiles by transition metal
complex-catalyzed cleavage is known, and these compounds, as well as hydride donors
(e.g., formic acid), are used as allyl scavengers16,17. This unexpected result can be ex-
plained by transformation resembling to the intramolecular Leuckart–Wallach reaction
as shown in Scheme 1. The reaction of compound 8 with hydrazine or hydrazine ace-
tate leads, besides cleavage of the phthalimido group, also to isomerization of the allyl
protecting group to propenyl group under formation of A. The subsequent intramolecu-
lar addition of free NH2-group to vinyl ether system of the propenyl group affords
cyclic intermediate B. The action of formic acid added during workup of the reaction
mixture opens the cyclic compound B to give ion C. Reduction of C by formic acid
affords the N-propyl derivative 9. We consider an alternative intermolecular course of
this reaction based on the cleavage of the propenyl group of the intermediate A by
formic acid to give benzyl 2-acetamido-4-O-(2-amino-3,4,6-tri-O-benzyl-2-deoxy-β-D-
glucopyranosyl)-6-O-benzyl-2-deoxy-α-D-glucopyranoside (10) and propionaldehyde
and their subsequent Leuckart–Wallach reaction to compound 9 to be less probable,
owing to a high yield of propyl derivative 9. The presence of 10 in the reaction mixture
was not observed. Compound 10 was prepared from benzyl 2-acetamido-3-O-allyl-4-O-
(2-amino-3,4,6-tri-O-benzyl-2-deoxy-β-D-glucopyranosyl)-6-O-benzyl-2-deoxy-α-D-
glucopyranoside (11) by cleavage of the allyl protecting group via its catalytic isomeri-
zation to propenyl group by Wilkinson’s catalyst [(Ph3P)3RhCl] followed by acidic
hydrolysis. Amine 11 was prepared by two-steps reductive dephthaloylation11,18 of
compound 8. The phthalimido group was reduced with NaBH4 to the corresponding
2-(hydroxymethyl)benzamide, which was in the second step cleaved by acetic acid at
85 °C.
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

Synthesis of Analogs of GMDP
581
N-Acylation of amine 11 with acetic anhydride or stearoyl chloride in a mixture of
pyridine and dichloromethane afforded benzyl 2-acetamido-4-O-(2-acetamido-3,4,6-tri-
O-benzyl-2-deoxy-β-D-glucopyranosyl)-3-O-allyl-6-O-benzyl-2-deoxy-α-D-glucopyra
noside (12a, ref.1) or benzyl 2-acetamido-3-O-allyl-4-O-(3,4,6-tri-O-benzyl-2-deoxy-2-
stearoylamino-β-D-glucopyranosyl)-6-O-benzyl-2-deoxy-α-D-glucopyranoside (12b,
ref.3), respectively. These compounds can be transformed to the target glycopeptides,
OBzl
O
OBzl
O
OBzl
O
O
OBzl
NHAc
CH CH=CH
BzlO
NPht
2
2
8
OBzl
OBzl
O
OBzl
OBzl
O
OBzl
OBzl
O
O
O
O
O
O
OBzl
NHAc
CH=CHCH
OBzl
NHAc
BzlO
BzlO
NH
2
HN
CHCH CH
2
2
3
A
B
OBzl
O
OBzl
O
OBzl
OH
+
O
+
CH CH CHO
3 2
+ H
OBzl
NHAc
BzlO
NH
2
+
10
+ H
- H O
2
OBzl
O
OBzl
OBzl
O
OBzl
O
OBzl
OBzl
O
+
- H
OH
3
OH
O
O
OBzl
NHAc
OBzl
NHAc
BzlO
BzlO
N=CHCH CH
2
HN
CHCH CH
2 3
D
C
OBzl
O
OBzl
O
OBzl
OH
O
OBzl
NHAc
BzlO
HN CH CH CH
2
2
3
9
SCHEME 1
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

582
Ledvina, Zyka, Jezek, Trnka, Saman:
(2-acetamido-2-deoxy-β-D-glucopyranosyl)-(1→4)-N-acetylnormuramoyl-L-2-amino-
butanoyl-D-isoglutamine (16a) and (2-deoxy-2-stearamido-β-D-glucopyranosyl)-
(1→4)-N-acetylnormuramoyl-L-2-aminobutanoyl-D-isoglutamine (16b), respectively,
in four steps, according to our earlier described procedure1,3: (i) cleavage of the allyl
protecting group, (ii) O-alkylation of the formed hydroxy derivatives 13a and 13b by
chloroacetic acid in the presence of sodium hydride, (iii) DCC-promoted condensation
of the 3-O-carboxymethyl derivatives 14a and 14b with H-L-Abu-D-isoGln(OBzl) and
(iv) hydrogenolysis of benzyl protecting groups from glycopeptides 15a and 15b.
EXPERIMENTAL
Melting points were determined on a Kofler block and are uncorrected. Optical rotations were
measured on a Perkin–Elmer 141 polarimeter at 22 °C. The IR spectra were recorded on a Bruker
IFS 88 (FTIR) spectrometer, wavenumbers are given in cm–1. NMR spectra were recorded with a
Varian UNITY-500 spectrometer in the FT mode at 499.8 MHz (1H) and at 125.6 MHz (13C) in
deuteriochloroform, using tetramethylsilane as internal standard for the 1H NMR spectrum and
deuteriochloroform (δ 77.0) as standards for 13C NMR spectrum. Chemical shifts are given in ppm
(δ-scale) and coupling constants (J) in Hz. For uninterchangeable assignment of signals in 13C NMR
spectra of compounds 8, 10, 11 and 12a, the heterocorrelated 2D NMR spectra were measured by the
HMQC technique using the standard pulse sequence delivered by the producer of the spectrometer.
Following set of parameters was used: spectral width in both f1 and f2 dimensions 4 500 Hz and 17 000 Hz,
respectively, number of scans 32, number of increments in f1 dimension 256, recycle delay 1 s, acquisi-
1
tion time 0.2 s, 90° pulse for H was 22.5 µs, data matrix for processing 2 048 × 2 048 datapoints,
for processing no weighting function was used. Positive-ion FAB mass spectra were measured on a
BEqG geometry mass spectrometer ZAB-EQ (VG Analytical, Manchester, U.K.), using an M-Scan
FAB gun (Xe, energy 8 keV) at an accelerating voltage of 8 kV. Samples were dissolved in chloro-
form or methanol, and the mixture glycerol–thioglycerol or 3-nitrobenzyl alcohol was used as matrix.
Thin-layer chromatography (TLC) was performed on Silufol UV254 sheets, and column chromato-
graphy on silica gel Silpearl (both Kavalier, Votice, Czech Republic). Analytical RP HPLC was per-
formed with a Spectra-Physics 8700 apparatus (Darmstadt, Germany) equipped with a column (250 × 4 mm)
filled with Separon SGX-RPS (C18), particle size 10 nm (Tessek, Prague). Preparative RP HPLC was
performed with a Knauer apparatus (Bad Homburg, Germany) equipped with a column (250 × 10 mm)
filled with Separon SGX-RPS, particle size 10 nm (Tessek, Prague). Solutions were evaporated on
rotatory vacuum evaporator. Analytical samples were dried at 6.5 Pa and 25 °C for 8 h.
Dichloromethane was distilled from phosphorus pentoxide and stored over molecular sieves 4A.
Silver trifluoromethanesulfonate was recrystallized from toluene.
1,3,4,6-Tetra-O-acetyl-2-deoxy-2-phthalimido-β-D-glucopyranose (2)
Solution of 1,3,4,6-tetra-O-acetyl-2-amino-2-deoxy-β-D-glucopyranose8 (1, 173.7 g, 0.5 mol),
phthalic anhydride (75.54 g, 0.51 mol) and 4-dimethylaminopyridine (10.0 g, 81.8 mmol) in dichlo-
romethane (1 500 ml) was stirred at ambient temperature until of the starting amine 1 disappeared;
the reaction was monitored by TLC in ethyl acetate–chloroform–formic acid (20 : 10 : 1). Acetic an-
hydride was added (300 ml, 3.18 mol) and the mixture was allowed to stand overnight; the reaction
was monitored by TLC in ethyl acetate–toluene (1 : 1). Under stirring, ethanol (200 ml) was added
and, after 2 h at ambient temperature the solvents were evaporated and the residue was codistilled
with toluene (3 × 500 ml) to give a syrup, which crystallized on addition of ethanol and standing
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

Synthesis of Analogs of GMDP
583
overnight at +3 °C, to yield crude 2 (195.4 g, 82%); m.p. 72–75 °C. Recrystallization of the product
from ethanol afforded 175.8 g (75%) of compound 2; m.p. 79–81 °C, [α]D +62° (c 0.56, chloroform);
ref.9: m.p. 69–71 °C; ref.11: m.p. 74–75 °C, [α]D +71° (c 1.3, chloroform). IR spectrum (tetrachloro-
methane): 3 088, 3 062, 3 028 (C–H, Pht); 1 783, 1 725 (C=O, Pht); 1759 (C=O, Ac); 1 615, 1 592,
1 470, 1 384 (arom. ring). 1H NMR spectrum: 7.86–7.88 m, 7.75–7.77 m, 4 H (H-arom.); 6.52 d,
1 H, J = 8.8 (H-1); 5.89 dd, 1 H, J = 9.0, 10.5 (H-3); 5.22 dd, 1 H, J = 9.0, 10.3 (H-4); 4.47 dd, 1 H,
J = 8.8, 10.5 (H-2); 4.37 dd, 1 H, J = 4.3, 12.4 (H-6a); 4.15 dd, 1 H, J = 2.2, 12.4 (H-6b); 4.03 ddd,
1 H, J = 2.2, 4.3, 10.3 (H-5); 1.87 s, 2.00 s, 2.05 s, 2.12 s, 4 × 3 H (3 × CH3CO). 13C NMR spectrum:
170.7 s, 170.1 s, 169.5 s, 168.7 s (4 × CH3COO); 167.4 s, 2 × C (2 × C=O, Pht); 134.5 d, 2 × C,
131.2 s, 2 × C, 123.8 d, 3 × 2 C (Pht); 89.8 d (C-1); 72.6 d (C-5); 70.5 d (C-3); 68.3 d (C-4); 61.5 t
(C-6); 53.5 d (C-2); 20.7 q, 20.7 q, 20.6 q, 20.4 q (4 × COCH3). For C22H23NO11 calculated: relative
molecular mass 477.4, monoisotopic mass 477.1. FAB MS, m/z: 478.3 [M + H]+, 500.3 [M + Na]+.
For C22H23NO11 (477.4) calculated: 55.34% C, 4.85% H, 2.93% N; found: 55.19% C, 4.85% H,
2.85% N.
Ethyl 3,4,6-Tri-O-acetyl-2-deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (3)
To a stirred solution of compound 2 (167 g, 0.35 mol) and ethanethiol (66 ml, 0.89 mol) in dry di-
chloromethane (1 700 ml) in the presence of powdered molecular sieves 4A (150 g) at 0 °C titanium
tetrachloride (50 ml, 0.46 mol) was slowly added. After 1 h stirring at ambient temperature the reac-
tion mixture was filtered through a layer of cellite and the cellite was washed with dichloromethane
(1 500 ml). Collected filtrate was washed with 1 M H2SO4 (800 ml), saturated solution of sodium
hydrogen carbonate till neutral pH, and water (800 ml), dried over anhydrous magnesium sulfate, and
the solvent was evaporated. The residue was crystallized from a mixture of ethyl acetate and petroleum
ether. Yield 121 g (72%) of compound 3; m.p. 115–118 °C, [α]D +46° (c 0.8, dichloromethane);
ref.7: m.p. 118–119 °C, [α]D +44° (c 0.8, dichloromethane). For C22H25NO9S calculated: relative
molecular mass 479.5, monoisotopic mass 479.1. FAB MS, m/z: 480.1 [M + H]+, 502.1 [M + Na]+.
For C22H25NO9S (479.5) calculated: 55.10% C, 5.25% H, 2.92% N, 6.68% S; found: 54.94% C,
5.11% H, 2.98% N, 6.46%S.
Ethyl 2-Deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (4)
A suspension of compound 3 (128 g, 0.27 mol; dried 6 h at room temperature and 1.32 Pa) in 0.01 M
CH3ONa in methanol (2 500 ml) was stirred at room temperature for 2 h; during this time the sus-
pension dissolved. The solution was allowed to stand overnight at 3 °C and neutralized by addition
of Dowex 50 (pyridinium form). The ion exchanger was filtered off, washed with methanol and the
filtrate was concentrated in vacuo. Crystallization of the residue from dichloromethane afforded
85.5 g (90%) of product 4; m.p. 169–170 °C, [α]D +9° (c 0.6, methanol); compound 4 is described
in ref.7 as a non characterized intermediate. IR spectrum (KBr): 3 494, 3 458, 3 361 (O–H, bonded);
3 109, 3 051, 3 026 (C–H, Pht); 1 767, 1 754, 1 748, 1 700, 1 673 (C=O, Pht); 1 611, 1 466, 1 394, 1 383
(arom. ring). For C16H19NO6S calculated: relative molecular mass 353.4, monoisotopic mass 353.1.
FAB MS, m/z: 376.0 [M + Na]+. For C16H19NO6S (353.4) calculated: 54.38% C, 5.41% H, 3.96% N,
9.07% S; found: 54.55% C, 5.24% H, 3.95% N, 9.04% S.
Ethyl 3,4,6-Tri-O-benzyl-2-deoxy-2-phthalimido-1-thio-β-D-glucopyranoside (5)
A solution of compound 4 (42.4 g, 120 mmol) and benzyl bromide (85.6 ml, 720 mmol) in dry N,N-
dimethylformamide (600 ml) was slowly added to a stirred 60% suspension of NaH in mineral oil
(28.8 g, 720 mmol) at 0 °C during 30 min under nitrogen, and the mixture was stirred at ambient
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temperature overnight. Acetic anhydride (350 ml, 3.7 mol) was added and stirring continued for 3 h.
The solution was poured under stirring and ice-bath cooling to a saturated sodium hydrogen carbo-
nate solution (2 l). The product was extracted with toluene (3 × 600 ml), the extracts were dried over
anhydrous magnesium sulfate, and concentrated in vacuo. Chromatography of the residue on a silica
gel column (1 000 g) in toluene–ethyl acetate (20 : 1) afforded 65.5 g (88%) of syrupy product,
which was crystallized from toluene–petroleum ether. Yield 54.1 g (72%) of compound 5; m.p. 103–104 °C,
[α]D +63° (c 0.4, chloroform). IR spectrum (tetrachloromethane): 3 089, 3 066, 3 032 (C–H, Bzl, Pht);
1 778, 1 718 (C=O, Pht); 1 612, 1 588, 1 497, 1 470, 1 454, 1 387 (arom. ring Bzl, Pht). 1H NMR
spectrum: 6.83–7.80 m, 3 × 5 H (H-arom., Bzl); 6.83–7.50 m, 4 H (H-arom., Pht); 5.26 d, 1 H, J =
10.5 (H-1); 4.84 d, 1 H, J = 10.5 (CH2–Ph); 4.79 d, 1H, J = 12.0 (CH2–Ph); 4.65 d, 1H, J = 12.0
(CH2–Ph); 4.65 d, 1 H, J = 10.5 (CH2–Ph); 4.58 d, 1 H, J = 12.0 (CH2–Ph); 4.45 d, J = 12.0 (CH2–Ph);
4.39 dd, 1 H, J = 8.7, 10.3 (H-3); 4.26 t, 1 H, J = 10.3 (H-2); 3.81 dd, 1 H, J = 2.4, 11.0 (H-6b);
3.78 dd, 1 H, J = 4.1, 11.0 (H-6a); 3.78 dd, 1 H, J = 8.5, 10.0 (H-4); 3.68 ddd, 1 H, J = 2.4, 4.1,
10.0 (H-5); 2.69 dq, 1 H, J = 7.3, 12.7 (CH3CHHS); 2.60 dq, 1 H, J = 7.3, 12.7 (CH3CHHS); 1.18 t,
3 H, J = 7.3 (CH3CH2S). 13C NMR spectrum: 168.0 s, 167.5 s, 138.3 s, 138.0 s (2 × C), 133.8 d,
133.7 d, 132.0 s, 131.7 s, 128.9 d (2 × C), 128.4 d (2 × C), 128.1 d (3 × C), 127.9 d (4 × C), 127.8 d,
127.7 d, 127.6 d, 127.3 d, 123.4 d, 123.2 d (arom. C, Bzl, Pht); 81.0 d (C-1); 80.4 d (C-3); 79.6 d
(C-4); 79.4 d (C-5); 75.0 t, 74.9 t, 73.4 t (CH2–Ph); 68.9 t (C-6); 55.0 d (C-2); 23.86 t (CH3CH2S);
14.9 q (CH3CH2S). For C37H37NO6S calculated: relative molecular mass 623.8, monoisotopic mass
623.2. FAB MS, m/z: 624.1 [M + H]+, 646.1 [M + Na]+. For C37H37NO6S (623.8) calculated: 71.24% C,
5.97% H, 2.24% N, 5.14% S; found: 71.05% C, 5.99% H, 2.18% N, 5.30% S.
Benzyl 2-Acetamido-3-O-allyl-6-O-benzyl-4-O-(3,4,6-tri-O-benzyl-2-deoxy-2-phthalimido-
β-D-glucopyranosyl)-2-deoxy-α-D-glucopyranoside (8)
Method A. A mixture of compound 5 (312 mg, 0.5 mmol), benzyl 2-acetamido-3-O-allyl-6-O-
benzyl-2-deoxy-D-glucopyranoside (7, ref.1; 221 mg, 0.5 mmol) and molecular sieves 4A (0.8 g) was
dried in an apparatus with a septum, for 8 h at room temperature and 1.32 Pa. The apparatus was
flushed with argon (2 ×) and dry dichloromethane (10 ml) was added through the septum. After dis-
solution, the mixture was cooled to –15 °C and methyl triflate (226 µl, 2 mmol) was added under
stirring through the septum and the mixture was allowed to stand at –15 °C for 3 days. Triethylamine
(200 µl, 1.4 mmol) was added and after 5 min stirring, the mixture was filtered. The filtrate was
diluted with chloroform (70 ml) and the solution was washed with 1 M H2SO4, saturated NaHCO3
and water (3 × 20 ml each), dried over anhydrous magnesium sulfate, and the solvents were evapo-
rated. Chromatography of the residue on silica gel column (30 g) in toluene–ethyl acetate (2 : 1)
followed by lyophilization from benzene afforded 105 mg (21%) of product 8; m.p. 50–54 °C, [α]D
+63° (c 0.5, chloroform). IR spectrum (tetrachloromethane): 3 449 (NH, NHAc); 3 089, 3 066, 3 033
(C–H, Bzl, Pht, All); 1 779, 1 717 (C=O, Pht); 1 692 (amide I); 1 614, 1 587, 1 497, 1 469, 1 454, 1 388
1
(arom. ring, Bzl, Pht); 1 508 (amide II). H NMR spectrum: 6.88–7.80 m, 19 H (arom. H, Bzl, Pht);
5.77 dddd, 1 H, J = 4.6, 6.1, 10.5, 17.2 (H-2′′); 5.48 bd, 1 H, J = 9.3 (NHCOCH3); 5.29 d, 1 H, J =
8.6 (H-1′); 5.17 ddt, 1 H, J = 1.7, 1.7, 2.0, 17.2 (H-3a′′); 4.99 ddt, 1 H, J = 1.4, 1.4, 1.7, 10.5
(H-3b′′); 4.82 d, 1 H, J = 3.6 (H-1); 4.80 d, 1 H, J = 11.2 (CH2–Ph); 4.78 d, 1 H, J = 12.2 (CH2–Ph);
4.67 d, 1 H, J = 11.2 (CH2–Ph); 4.64 d, 1 H, J = 12.2 (CH2–Ph); 4.54 d, 1 H, J = 12.2 (CH2–Ph);
4.53 d, 1 H, J = 11.9 (CH2–Ph); 4.41 d, 1 H, J = 12.2 (CH2–Ph); 4.39 d, 1 H, J = 12.2 (CH2–Ph);
4.38 ddt, 1 H, J = 1.5, 1.5, 4.6, 13.1 (H-1a′′); 4.34 d, 1 H, 11.9 (CH2–Ph); 4.34 d, 1 H, J = 12.2
(CH2–Ph); 4.33 dd, 1 H, J = 8.8, 11.0 (H-3′); 4.16 ddd, 1 H, J = 3.6, 9.3, 10.5 (H-2); 4.14 dd, 1 H,
J = 8.6, 11.0 (H-2′); 4.01 dd, 1 H, J = 8.8, 9.8 (H-4); 3.99 ddt, 1 H, J = 1.5, 1.5, 6.1, 13.1 (H-1b′′);
3.86 dd, 1 H, J = 8.8, 9.8 (H-4′); 3.75 dd, 1 H, J = 3.2, 11.1 (H-6a′); 3.72 dd, 1 H, J = 2.2, 11.1
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Synthesis of Analogs of GMDP
585
(H-6b′); 3.52 dt, 1 H, J = 2.7, 2.7, 9.8 (H-5); 3.50 dd, 1 H, J = 8.8, 10.5 (H-3); 3.43 ddd, 1 H, J =
2.2, 3.2, 9.8 (H-5′); 3.34–3.38 m, 2 H (H-6a and H-6b); 1.91 s, 3 H (NHCOCH3). 13C NMR spec-
trum: 169.7 s (NHCOCH3); 168.0 s, 167.8 s, 138.3 s (2 × C), 138.1 s, 138.0 s, 137.1 s, 133.7 d,
131.8 s, 131.4 s, 128.4 d (4 × C), 128.3 d (3 × C), 128.2 d (3 × C), 128.0 d (2 × C), 127.9 d (3 × C),
127.7 d (2 × C), 127.6 d (3 × C), 127.4 d (2 × C), 127.3 d (3 × C), 127.2 d (2 × C), 123.2 d (arom. C,
Bzl, Pht); 135.5 d (C-2′′); 116.0 t (C-3′′); 97.1 d (C-1′); 96.7 d (C-1); 79.5 d (C-4′); 79.0 d (C-3′);
78.1 d (C-3); 74.8 t (2 × C) (CH2–Ph); 74.9 d (C-5′); 74.7 d (C-4); 73.3 t (CH2–Ph); 72.6 t (C-1′′);
72.6 t (CH2–Ph); 70.6 d (C-5); 69.5 t (CH2–Ph); 68.2 t (C-6′); 68.1 t (C-6); 56.6 d (C-2′); 52.1 d
(C-2); 23.3 q (NHCOCH3). For C60H62N2O12 calculated: relative molecular mass 1 003.2, monoiso-
topic mass 1 002.4. FAB MS, m/z: 1 003.4 [M + H]+. For C60H62N2O12 (1 003.2) calculated: 71.83% C,
6.22% H, 2.79% N; found: 71.63% C, 6.29% H, 2.77% N.
Method B. Compound 5 (18.7 g, 30 mmol) was dried in an apparatus with a septum at 1.32 Pa and
room temperature for 8 h. The apparatus was flushed with argone (2 ×) and dry dichloromethane
(100 ml) was added through the septum. After dissolution, the mixture was cooled to 0 °C and a
solution of 1 M bromine in dichloromethane (30 ml, 30 mmol) was added through the septum. The
mixture was stirred at 0° C for 45 min. In the same apparatus, the solvents were evaporated in vacuo
(water pump) with exclusion of moisture and the residue was coevaporated with dry toluene (3 × 70 ml)
at 133.3 Pa, added through the septum, and dried at 1.32 Pa for 1 h. The solid foam obtained was
dissolved by addition of dry dichloromethane (55 ml) through the septum and the resulting solution
of glycosylbromide 6 was immediately used for the condensation with 7.
A mixture of compound 7 (8.83 g, 20 mmol) and silver triflate (7.71 g, 30 mmol) was dried in
apparatus equipped with a septum at room temperature and 1.32 Pa for 6 h. The apparatus was
flushed with argone (2 ×) and dry dichloromethane (55 ml) was added through the septum. After
dissolution, the mixture was cooled to –45 °C and the solution of glycosyl donor 6 (see above) was
added through the septum under stirring during 1 h. The mixture was stirred for another 1 h at –45 °C
and 30 min at –20 °C. Pyridine (9 ml) was added at –20 °C and after warming to room temperature
the mixture was diluted with chloroform (300 ml) and filtered. The filtrate was washed with 0.5
M
HCl (3 × 200 ml), saturated solution of NaHCO3 (3 × 200 ml) and water (2 × 200 ml), dried over
anhydrous magnesium sulfate and evaporated. The residue was worked up by the same procedure as
given in A to give 14.5 g (72%) of compound 8, identical (m.p., [α]D, IR and NMR spectra) with
compound prepared by method A. For C60H62N2O12 (1 003.2) calculated: 71.83% C, 6.22% H, 2.79% N;
found: 71.70% C, 6.32% H, 2.69% N.
Benzyl 2-Acetamido-4-O-(3,4,6-tri-O-benzyl-2-deoxy-2-propylamino-β-D-glucopyranosyl)-
6-O-benzyl-2-deoxy-α-D-glucopyranoside (9)
Method A. Compound 8 (2.0 g, 2 mmol) was refluxed in a mixture of ethanol–hydrazine hydrate
(2 : 1, 50 ml) for 5 h. The mixture was concentrated in vacuo, coevaporated with toluene (3 × 30 ml)
and the residue was dissolved in small amount of methanol. The pH of the solution was adjusted to
5 with formic acid and the solution was poured on a column of Dowex 50 in the pyridinium form
(80 ml). The column was washed with methanol (600 ml) and the product was desorbed with 5%
solution of triethylamine in methanol (600 ml). The eluate was evaporated to give 1.46 g of solid
residue, which was crystallized from toluene–petroleum ether. Yield 1.05 g (60%) of compound 9;
m.p. 145–147 °C, [α]D +68° (c 0.4, chloroform). IR spectrum (tetrachloromethane): 3 452 (N–H,
amide); 3 090, 3 067, 3 032, 3 010 (C–H, Bzl); 1 691 (amide I); 1 608, 1 498, 1 454 (arom. ring, Bzl);
1 505 (amide II). 1H NMR spectrum: 7.16–7.36 m, 25 H (arom. H, Bzl); 5.54 d, 1 H, J = 9.2
(NHCOCH3); 4.94 d, 1 H, J = 3.8 (H-1); 4.93 d, 1 H, J = 11.1 (CH2–Ph); 4.75 d, 1 H, J = 10.9
(CH2–Ph); 4.68 d, 1 H, J = 10.9 (CH2–Ph); 4.68 d, 1 H, J = 11.7 (CH2–Ph); 4.68 d, 1 H, J = 11.1
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586
Ledvina, Zyka, Jezek, Trnka, Saman:
(CH2–Ph); 4.66 d, 1 H, J = 12.1 (CH2–Ph); 4.60 d, 1 H, J = 11.7 (CH2–Ph); 4.53 d, 1 H, J = 12.0
(CH2–Ph); 4.49 d, 1 H, J = 12.1 (CH2–Ph); 4.45 d, 1 H, J = 12.0 (CH2–Ph); 4.28 d, 1 H, J = 8.0
(H-1′); 4.25 ddd, 1 H, J = 3.8, 9.2, 10.6 (H-2); 4.05 dd, 1 H, J = 8.9, 9.8 (H-4); 3.95 dd, 1 H, J =
3.6, 11.0 (H-6b); 3.79 ddd, 1 H, J = 1.8, 3.6, 9.8 (H-5); 3.77 dd, 1 H, J = 9.1, 9.7 (H-4′); 3.67 dd, 1 H,
J = 1.8, 11.0 (H-6a); 3.66–3.75 m, 2 H (H-6a′ and H-6b′); 3.53 dd, 1 H, J = 8.9, 10.6 (H-3); 3.36 dt,
2 H, J = 6.8, 6.8, 8.9 (NHCH2CH2CH3); 3.29 ddd, 1 H, J = 2.3, 3.3, 9.7 (H-5′); 3.28 dd, 1 H, J =
9.1, 9.8 (H-3′); 2.77 dd, 1 H, J = 8.0, 9.8 (H-2′); 1.95 s, 3 H (NHCOCH3);1.38–1.52 m, 2 H
(NHCH2CH2CH3); 0.78 t, 3 H, J = 7.3 (NHCH2CH2CH3). 13C NMR spectrum: 169.6 s (NHCOCH3);
138.5 s, 138.3 s, 138.2 s, 138.1 s, 137.3 s (5 × arom. C, Bzl); 128.5 d (4 × C), 128.4 d (3 × C), 128.3 d
(2 × C), 128.0 d (2 × C), 127.8 d (2 × C), 127.7 d (12 × C) (arom. C, Bzl); 102.9 d (C-1′); 97.1 d
(C-1); 85.1 d (C-3′); 78.4 d (C-4, C-4′); 75.1 d (C-5′); 74.6 t (CH2–Ph); 73.4 t (CH2–Ph); 73.3 t
(CH2–Ph); 73.0 t (CH2–Ph); 71.2 d (C-3); 70.0 d (C-5); 69.7 t (CH2–Ph); 68.8 t (C-6′); 68.3 t (C-6);
57.6 d (C-2′); 52.0 d (C-2); 23.4 q (NHCOCH3); 23.3 t (C-2′′); 17.7 t (C-1′′); 10.5 q (C-3′′). For
C52H62N2O10 calculated: relative molecular mass 875.1, monoisotopic mass 874.4. FAB MS, m/z:
875.3 [M + H]+. For C52H62N2O10 (875.1) calculated: 71.37% C, 7.14% H, 3.20% N; found: 71.28% C,
7.08% H, 3.23% N.
Method B: Compound 8 (1.0 g, 1 mmol) was heated in a solution of hydrazine acetate (1.84 g, 20 mmol)
in ethanol (18 ml) at 80 °C for 9 h. The mixture was concentrated in vacuo, coevaporated with toluene
(3 × 20 ml) and the residue was taken between chloroform (100 ml) and water (20 ml). The organic
layer was separated, washed with water (2 × 20 ml), dried over anhydrous magnesium sulfate and
evaporated. The residue was worked up, using the same procedure as in method A, to give 535 mg
(61%) of compound 9, identical (m.p., [α]D, IR and NMR spectra) with compound prepared by
method A. For C52H62N2O10 (875.1) calculated: 71.37% C, 7.14% H, 3.20% N; found: 71.21% C, 7.12% H,
3.19% N.
Benzyl 2-Acetamido-4-O-(2-amino-3,4,6-tri-O-benzyl-2-deoxy-β-D-glucopyranosyl)-6-O-benzyl-
2-deoxy-α-D-glucopyranoside (10)
A mixture of compound 11 (1.745 g, 2 mmol) and chlorotris(triphenylphosphine)rhodium(I) (200 mg,
0.22 mmol) was refluxed in a mixture of ethanol–toluene–water (7 : 3 : 1, 80 ml) for 10 h under stir-
ring. Formic acid was added (2 ml) and the mixture was refluxed for another 1 h. After cooling, the
mixture was filtered and the filtrate was evaporated. The residue was dissolved in a small amount of
methanol, pH of the solution was adjusted to 5 with formic acid and the solution was poured on a
column of Dowex 50 in the pyridinium form (60 ml). The column was washed with methanol (600 ml)
and the product desorbed with 5% triethylamine in methanol (600 ml). The eluate was evaporated
and the residue coevaporated with toluene (3 × 20 ml). Crystallization of the residue from a mixture
toluene–petroleum ether afforded 1.14 g (68%) of product 10; m.p. 170–175 °C, [α]D +60° (c 0.4,
chloroform). IR spectrum (tetrachloromethane): 3 455 (N–H, amide); 3 090, 3 066, 3 033, 3 008 (C–H,
Bzl); 1 688 (amide I); 1 648 (NH2); 1 608, 1 587, 1 498, 1 454 (arom. ring, Bzl); 1 507 (amide II). 1H NMR
spectrum: 7.26–7.35 m, 25 H (H arom., Bzl); 5.60 d, 1 H, J = 8.4 (NHCOCH3); 5.03 d, 1 H, J = 3.6
(H-1); 4.93 d, 1 H, J = 11.3 (CH2–Ph); 4.76 d, 1 H, J = 10.9 (CH2–Ph); 4.72 d, 1 H, J = 11.8
(CH2–Ph); 4.68 d, 1 H, J = 11.3 (CH2–Ph); 4.62 d, 1 H, J = 12.1 (CH2–Ph); 4.56 d, 1 H, J = 11.9
(CH2–Ph); 4.56 d, 1 H, J = 11.8 (CH2–Ph); 4.53 d, 1 H, J = 10.9 (CH2–Ph); 4.47 d, 1 H, J = 12.1
(CH2–Ph); 4.47 d, 1 H, J = 11.9 (CH2–Ph); 4.15 d, 1 H, J = 8.1 (H-1′); 4.13 ddd, 1 H, J = 3.6, 8.4,
10.6 (H-2); 3.86 ddd, 1 H, J = 2.0, 4.3, 9.9 (H-5); 3.78 dd, 1 H, J = 8.4, 10.6 (H-3); 3.77 dd, 1 H,
J = 4.3, 10.0 (H-6a); 3.70 dd, 1 H, J = 2.0, 10.0 (H-6b); 3.70 dd, 1 H, J = 2.3, 10.7 (H-6b′); 3.65 dd,
1 H, J = 5.1, 10.7 (H-6a′); 3.64 dd, 1 H, J = 8.4, 9.9 (H-4); 3.61 dd, 1 H, J = 9.0, 9.9 (H-4′); 3.49 ddd,
1 H, J = 2.3, 5.1, 9.9 (H-5′); 3.30 dd, 1 H, J = 9.0, 10.0 (H-3′); 2.78 dd, J = 8.1, 10.0 (H-2′); 1.97 s,
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Synthesis of Analogs of GMDP
587
3 H (NHCOCH3). 13C NMR spectrum: 170.1 s (NHCOCH3); 138.2 s (2 × C), 137.7 s (2 × C), 137.3 s,
128.6 d (2 × C), 128.5 d (4 × C), 128.4 d (5 × C), 127.9 d (10 × C), 127.8 d (2 × C), 127.7 d (2 × C)
(30 × C, arom. C, Bzl); 104.3 d (C-1′); 96.7 d (C-1); 84.7 d (C-3′); 81.3 d (C-4); 78.2 d (C-4′); 75.4 t
(CH2–Ph); 74.8 d (C-5′); 74.8 t (CH2–Ph); 73.5 t (CH2–Ph); 73.3 t (CH2–Ph); 70.3 d (C-3); 69.9 d
(C-5); 69.8 t (CH2–Ph); 68.7 t (2 × C) (C-6 and C-6′); 57.0 d (C-2′); 53.3 d (C-2); 23.4 q
(NHCOCH3). For C49H56N2O10 calculated: relative molecular mass 833.0, monoisotopic mass 832.4.
FAB MS, m/z: 833.5 [M + H]+. For C49H56N2O10 (833.0) calculated: 70.65% C, 6.77% H, 3.36% N;
found: 70.47%C, 6.80% H, 3.27% N.
Benzyl 2-Acetamido-3-O-allyl-4-O-(2-amino-3,4,6-tri-O-benzyl-2-deoxy-β-D-glucopyranosyl)-
6-O-benzyl-2-deoxy-α-D-glucopyranoside (11)
Sodium borohydride (3.03 g, 80 mmol) was gradually added during 2 h to a stired solution of com-
pound 8 (10.03 g, 10 mmol) in a mixture of propan-2-ol–water (6 : 1, 200 ml) at room temperature
and the stirring was continued for another 2 h. The solvents were evaporated and the residue was
coevaporated with toluene (3 × 60 ml), dissolved in chloroform (350 ml) and extracted with water
(3 × 100 ml). The organic phase was dried over anhydrous magnesium sulfate, filtered and evapo-
rated. The solid residue was dissolved in a mixture of toluene–acetic acid (6 : 1, 70 ml) and heated
at 85 °C for 5 h. The solvents were evaporated, the residue coevaporated with toluene and dissolved
in a small amount of methanol. The pH of the solution was adjusted to 5 with formic acid and the
solution was poured on a column of Dowex 50 in the pyridinium form (100 ml). The column was
washed with methanol (800 ml) and the product was desorbed with 5% solution of triethylamine in
methanol (800 ml). Evaporation of the eluate afforded 6.1 g (70%) of a solid residue which was
crystallized from a mixture of toluene and petroleum ether. Yield 4.9 g (56%) of compound 11; m.p.
139 °C, [α]D +63° (c 0.2, chloroform). IR spectrum (tetrachloromethane): 3 451 (N–H, amide); 3 395,
1 646 (NH2); 3 090, 3 067, 3 032 (C–H, Bzl); 1 691 (amide I); 1 607, 1 498, 1 454 (arom. ring, Bzl);
1
1 506 (amide II). H NMR spectrum: 7.24–7.35 m, 25 H (arom. H, Bzl); 5.80 dddd, 1 H, J = 5.1, 6.1,
10.6, 17.0 (H-2′′); 5.51 d, 1 H, J = 9.3 (NHCOCH3); 5.15 dq, 1 H, J = 3 × 1.7, 17.0 (H-3a′′); 5.00 ddt,
1 H, J = 1.2, 1.2, 2.0, 10.6 (H-3b′′); 4.96 d, 1 H, J = 3.7 (H-1); 4.75 d, 1 H, J = 11.7 (CH2–Ph); 4.68 d,
1 H, J = 12.2 (CH2–Ph); 4.68 d, 1 H, J = 12.0 (CH2–Ph); 4.68 d, 1 H, J = 11.7 (CH2–Ph); 4.66 d, 1 H,
J = 12.0 (CH2–Ph); 4.60 d, 1 H, J = 11.7 (CH2–Ph); 4.59 d, 1 H, J = 12.0 (CH2–Ph); 4.53 d, 1 H, J =
12.0 (CH2–Ph); 4.50 d, 1 H, J = 12.2 (CH2–Ph); 4.46 d, 1 H, J = 11.7 (CH2–Ph); 4.41 ddt, 1 H, J =
1.5, 1.5, 5.1, 12.9 (H-1a′′); 4.27 d, 1 H, J = 7.8 (H-1′); 4.23 ddd, 1 H, J = 3.7, 9.0, 10.5 (H-2); 4.05 dd,
1 H, J = 8.9, 9.9 (H-4); 4.00 ddt, 1 H, J = 1.5, 1.5, 6.1, 12.9 (H-1b′′); 3.95 dd, 1 H, J = 3.7, 11.0
(H-6a′); 3.78 ddd, 1 H, J = 1.7, 3.7, 9.8 (H-5′); 3.71–3.74 m, 2 H (H-6); 3.70 t, 1 H, J = 9.5 (H-3′);
3.67 dd, 1 H, J = 1.7, 11.0 (H-6b′); 3.56 dd, 1 H, J = 8.9, 10.5 (H-3); 3.29 dt, 1 H, J = 2.8, 2.8, 9.9
(H-5); 3.28 bt, 1 H, J = 9.5 (H-4′); 2.77 dd, 1 H, J = 7.8, 10.0 (H-2); 1.94 s, 3 H (NHCOCH3). 13C NMR
spectrum: 169.7 s (NHCOCH3); 138.6 s, 138.4 s, 138.3 s, 138.2 s, 137.3 s (Ph); 135.8 d (C-2′′);
128.6 d, 128.5 d (2 × C), 128.4 d (3 × C), 128.3 d (2 × C), 128.1 d (3 × C), 127.8 d (6 × C), 127.7 d
(7 × C), 127.5 d (Ph); 115.9 t (C-2′′); 103.1 d (C-1′); 97.1 d (C-1); 85.0 d (C-5); 78.4 d (C-3); 78.5 d
(C-3′); 75.8 d (C-4); 75.2 t (CH2Ph); 75.2 d (C-4′); 74.6 t (CH2Ph); 73.5 t (CH2Ph); 73.3 t (CH2Ph);
72.5 t (C-1′′); 71.2 d (C-5′); 69.8 t (CH2Ph); 68.8 t (C-6); 68.3 t (C-6′); 57.7 d (C-2′); 52.2 d (C-2);
23.3 q (NHCOCH3). For C52H60N2O10 calculated: relative molecular mass 873.1, monoisotopic mass
872.4. FAB MS, m/z: 873.4 [M + H]+. For C52H60N2O10 (873.1) calculated: 71.53% C, 6.92% H,
3.20% N; found: 71.48% C, 7.00% H, 3.25% N.
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

588
Ledvina, Zyka, Jezek, Trnka, Saman:
Benzyl 2-Acetamido-4-O-(2-acetamido-3,4,6-tri-O-benzyl-2-deoxy-β-D-glucopyranosyl)-3-O-allyl-
6-O-benzyl-2-deoxy-α-D-glucopyranoside (12a)
To a stirred solution of compound 11 (2.62 g, 3.0 mmol) in a mixture of dichloromethane and py-
ridine (20 : 1, 42 ml) acetic acid anhydride (0.6 ml, 6.2 mmol) was slowly added and the mixture
was stirred for 30 min at 0 °C and another 30 min at room temperature. The solvents were evapo-
rated and the residue was coevaporated with toluene (3 × 15 ml). Column chromatography on silica
gel (50 g) in chloroform–ethyl acetate (2 : 1) yielded 2.0 g (73%) of crystalline compound 12a; m.p.
208–214 °C (dec.), [α]D +54° (c 0.4, chloroform); ref.1: m.p. 205–209 °C (dec.), [α]D +53° (c 0.2,
chloroform). 1H NMR spectrum: 7.24–7.37 m, 25 H (arom. H, Bzl); 5.77 dddd, 1 H, J = 4.8, 6.2,
10.5, 17.1 (H-2′′); 5.50 d, 2 H, J = 9.2 (2 × NHCOCH3); 5.14 dq, 1 H, J = 3 × 1.8, 17.1 (H-3a′′);
4.99 ddt, 1 H, J = 1.5, 1.5, 1.9, 10.5 (H-3b′′); 4.92 d, 1 H, J = 3.9 (H-1); 4.79 d, 1 H, J = 11.5
(CH2–Ph); 4.75 d, 1 H, J = 11.9 (CH2–Ph); 4.74 d, 1 H, J = 12.1 (CH2–Ph); 4.66 d, 1 H, J = 11.9
(CH2–Ph); 4.58 d, 1 H, J = 11.9 (CH2–Ph); 4.58 d, 1 H, J = 12.1 (CH2–Ph); 4.57 d, 1 H, J = 11.5
(CH2–Ph); 4.51 d, 1 H, J = 11.9 (CH2–Ph); 4.49 d, 1 H, J = 8.1 (H-1′); 4.45 d, 1 H, J = 12.1 (CH2–Ph);
4.41 d, 1 H, J = 11.9 (CH2–Ph); 4.41 ddt, 1 H, J = 1.4, 1.4, 4.8, 13.2 (H-1a′′); 4.19 ddd, 1 H, J =
3.9, 9.2, 10.6 (H-2); 3.96 ddt, 1 H, J = 1.4, 1.4, 6.2, 13.2 (H-1b′′); 3.89 dd, 1 H, J = 8.6, 9.9 (H-4);
3.74 dd, 1 H, J = 2.2, 10.8 (H-6a′); 3.70 dd, 1 H, J = 3.6, 10.8 (H-6b′); 3.67–3.70 m, 3 H (H-5, H-6a
and H-2′); 3.63 dd, 1 H, J = 8.8, 9.7 (H-4′); 3.54 dd, 1 H, J = 8.8, 9.7 (H-3′); 3.50 dd, 1 H, J = 8.6,
10.6 (H-3); 3.49 dd, 1 H, J = 2.4, 10.8 (H-6b); 3.35 ddd, 1 H, J = 2.2, 3.6, 9.7 (H-5′); 1.93 s, 3 H, 1.69 s,
3 H (2 × NHCOCH3). 13C NMR spectrum: 169.9 s (NHCOCH3); 169.7 s (NHCOCH3); 138.5 s, 138.3 s,
138.2 s (2 × C), 137.3 s (Ph); 135.8 d (C-2′′), 128.7 d (2 × C), 128.6 d (2 × C), 128.5 d (2 × C), 128.4 d
(4 × C), 128.3 d (2 × C), 128.1 d (2 × C), 128.0 d (3 × C), 127.9 d, 127.8 d (3 × C), 127.7 d, 127.6 d
(2 × C), 127.5 (Ph); 115.9 t (C-3′′); 100.2 d (C-1′); 97.0 d (C-1); 81.8 d (C-3′); 78.5 d (C-4′); 78.4 d
(C-3); 76.7 d (C-4); 74.8 d (C-5); 74.6 t (CH2–Ph); 74.5 t (CH2–Ph); 73.6 t (CH2–Ph); 73.4 t (CH2–Ph);
73.0 t (C-1′′); 70.7 d (C-5); 69.8 t (CH2–Ph); 68.8 t (C-6′); 68.1 t (C-6); 56.4 d (C-2′); 52.2 d (C-2);
23.4 q (NHCOCH3); 23.3 q (NHCOCH3). For C54H62N2O11 calculated: relative molecular mass
915.1, monoisotopic mass 914.4. FAB MS, m/z: 915.5 [M + H]+. For C54H62N2O11 (915.1) calcu-
lated: 70.89% C, 6.82% H, 3.06% N; found: 70.67% C, 6.86% H, 2.96% N.
Benzyl 2-Acetamido-3-O-allyl-6-O-benzyl-4-O-(3,4,6-tri-O-benzyl-2-deoxy-2-stearamido-
β-D-glucopyranosyl)-2-deoxy-α-D-glucopyranoside (12b)
To a stirred solution of compound 11 (5.24 g, 6.0 mmol) in a mixture of dichloromethane and py-
ridine (4 : 1, 70 ml), stearoyl chloride (3.03 g, 10 mmol) was added. After 5 h stirring at room tem-
perature, methanol (3 ml) was added and the mixture was stirred for 1 h. The solvents were
evaporated and the solid residue was extracted with petroleum ether (3 × 70 ml) to remove methyl
stearate. Chromatography of the residue on a silica gel column (120 g) in chloroform–ethyl acetate
(3 : 1) and concentration in vacuo yielded 5.8 g (85%) of solid product 12b, [α]D +43° (c 0.5, chlo-
roform); ref.3: [α]D +43° (c 0.4, chloroform). For C70H94N2O11 calculated: relative molecular mass 1 139.5,
monoisotopic mass 1 138.7. FAB MS, m/z: 1 139.8 [M + H]+. For C70H94N2O11 (1 139.5) calculated:
73.78% C, 8.31% H, 2.45% N; found: 72.52% C, 8.37% H, 2.39% N.
The authors would like to express their thanks to Dr V. Pechanec for elemental analyses and to Dr K. Ubik
for FAB MS spectra. Part of this work was supported by grant No. 203/96/0309 of the Grant Agency of
the Czech Republic.
Collect. Czech. Chem. Commun. (Vol. 63) (1998)

Synthesis of Analogs of GMDP
589
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