Synthesis of positively charged galactose-bearing surfactants

Article abstract of DOI:10.1134/S1068162010050146

An approach to the synthesis of cationic carbohydrate surfactants with potential antimicrobial and transfecting activities is proposed.

Full text of DOI:10.1134/S1068162010050146

ISSN 1068ꢀ1620, Russian Journal of Bioorganic Chemistry, 2010, Vol. 36, No. 5, pp. 657–662. © Pleiades Publishing, Ltd., 2010.
Original Russian Text © N.G. Morozova, M.A. Maslov, V.V. Myagchenkov, G.A. Serebrennikova, 2010, published in Bioorganicheskaya Khimiya, 2010, Vol. 36, No. 5, pp. 714–
720.
Synthesis of Positively Charged GalactoseꢀBearing Surfactants
N. G. Morozova1, M. A. Maslov, V. V. Myagchenkov, and G. A. Serebrennikova
Lomonosov State Academy of Fine Chemical Technology, pr. Vernadskogo 86, Moscow, 119571 Russia
Received January 1, 2010; in final form, March 31, 2010
Abstract—An approach to the synthesis of cationic carbohydrate surfactants with potential antimicrobial and
transfecting activities is proposed.
Key words: antimicrobial activity, cationic surfactants, galactose, Fukuyama reaction, transfection
DOI: 10.1134/S1068162010050146
2 1
INTRODUCTION
acyclic form) doubled the transfection activity as comꢀ
pared with surfactants containing two hydroxyl groups
[24].
Positively charged surfactants are widely used as
antiseptics and disinfectants [1]. As surfaceꢀactive
compounds, they interact with cell membrane phosꢀ
pholipids and damage membrane structures, causing
cell lysis [2].2 When added to bacterial cells or pathoꢀ
gen fungi, cationic surfactants, such as dioctadeꢀ
cyldimethylammonium bromide (DDAB) and hexaꢀ
decyltrimethylammonium bromide (CTAB), reverse
the cell membrane charge from negative to positive
and, thus, bring about cell death [3]. It was assumed
that the introduction of carbohydrate residues in the
structure of cationic surfactants could improve their
biocompatibility with cells and tissues [4].
The in vivo increased efficacy of nucleic acid delivꢀ
ery could be achieved if transport systems were to bear
specific ligands providing highly affine binding to speꢀ
cific receptors of target cells. It is known that hepatoꢀ
cyte membranes contain surfaceꢀexposed glycoproꢀ
tein receptors that allow the cell to recognize, immoꢀ
bilize, and swallow molecules bearing galactose
residues [25]. This property was used for the directed
delivery of nucleic acids into hepatocytes using galacꢀ
toseꢀcontaining cationic surfactants [26, 27]. Thus,
the search for biodegraded amphiphils among carboꢀ
hydrateꢀcontaining surfactants would provide an
extension of the spectrum of compounds with potenꢀ
tial antimicrobial activity and the development of
transport systems for the directed delivery of nucleic
acids to the target cells.
Due to their accessibility and low price, cationic
surfactants are also used in gene therapy as delivery
systems of therapeutic nucleic acids to eukaryotic cells
[5, 6]. Cationic surfactants protect nucleic acids from
enzymatic degradation by forming complexes with
them and support their transfer via cell membranes,
thus initiating endocytosis [7]. A large number of surꢀ
factants serve for the insertion of genetic constructs in
mammalian [5, 8, 9] and plant [10] cells. Particularly,
DDAB and dioleoyl dimethylammonium chloride
(DODAC) were used as mediators for the delivery of
plasmid DNA and antisense oligonucleotides in in
vitro [11–17] and in vivo [18, 19] experiments, as well
as in studies of the mechanism of DNA–cationic lipid
complex formation [20–22]. The replacement of two
DDAB methyl groups by 2ꢀhydroxyethyl groups
increased the transfection efficacy as compared with
both DDAB and the commercial transfection agent
lipofectamine (Lipofectamine, Invitrogen) [23]. The
introduction into the cationic surfactant structure of
four hydroxyl groups within arabinose or xylose (in the
RESULTS AND DISCUSSION
Earlier, we synthesized a series of cationic carbohyꢀ
drate amphiphils differing in the position of the posiꢀ
tively charged group and the spacer between the carꢀ
bohydrate unit and the hydrophobic domain [28, 29].
As a continuation of these studies, we report herein the
synthesis of cationic carbohydrate surfactants with
potential biological activity.
We used a tetradecyl substituent as a hydrophobic
domain and an ammonium fragment as a cationic
domain for the synthesis of carbohydrate surfactants.
Carbohydrate residues (galactose and lactose) were
introduced in the surfactant molecule using a hexamethꢀ
ylene spacer. Earlier, for the synthesis of similar comꢀ
pounds, an approach was proposed based on the couꢀ
pling of a trichloroacetamidate glucose derivative with
2ꢀbromoethanol followed by the quaternization of variꢀ
ous tertiary amines with the resulting 2ꢀbromoethylgluꢀ
coside [30]. Although this method is attractive due to the
small number of stages, its major disadvantage is the lack
1
Corresponding author; phone: +7 (495) 936ꢀ8903; fax: +7 (495)
936ꢀ8901; eꢀmail: ngmoroz@mail.ru.
2
Abbreviations: DDAB, dioctadecyldimethylammonium broꢀ
mide; CTAB, hexadecyltrimethylammonium bromide;
DODAC, dioleoyl dimethylammonium chloride.
657

658
MOROZOVA et al.
of a universal character because of the limited choice of approach based on the Fukuyama reaction [31, 32],
starting materials. A more universal synthetic scheme was namely, the interaction of 2ꢀnitrobenzenesulfonyl
described quite recently, which provided length variaꢀ amides with alkyl halogenides followed by the removal of
tions of the spacer joining a positively charged group and the 2ꢀnitrobenzenesulfonyl group. The resulting secondꢀ
a carbohydrate residue [27]. However, the total yield of ary amines can be easily subjected to quaternization with
the finished products was only 3%. We proposed another various alkyl halogenides at the next stages.
2ꢀNO ꢀC H ꢀSO Cl, Et N
2
6
4
2
3
HO
NH
O S O
HO
NH2
4
4
(I)
NO2
(II)
OAc
R
HO
N C14H29
4
O
R1
AcO
O S O
C H Br, K CO
14 29 2
3
NO2
AcO
Br
(III)
(
(
IVа), R = OAc, R1 = H
IVb), R = H, R1 = Oꢀ
ꢀDꢀGal(Ac)4
β
OAc
O
OAc
R
R
CdCO3
+
R1
O
O
R1
AcO
O
N C14H29
O S O
4
+
AcO
O
N C14H29
4
AcO
AcO
O S O
O
N C14H29
O S O
NO2
4
(
VII)
NO2
NO2
(
(
Vа), R = OAc, R1 = H
Vb), R = H, R1 =
Oꢀ ꢀDꢀGal(Ac)4
β
(
(
VIа), R = OAc, R1 = H
VIb), R = H, R1 =
OꢀβꢀDꢀGal(Ac)4
MeONa/MeOH
OH
OH
R
R
PhSH, K CO
2
3
R1
HO
R1
HO
O
O
O
N C14H29
O
N C14H29
H
4
4
HO
HO
IXа), R = OH, R1 = H
IXb), R = H, R1 =
Oꢀ ꢀDꢀGal
O S O
(
(
NO2
β
(
(
VIIIа), R = OH, R1 = H
VIIIb), R = H, R1 =
Oꢀ ꢀDꢀGal
β
OH
R
I–
CH I, Cs CO
3
R1
HO
O
+ Me
N C14H29
Me
3
2
O
4
HO
(
(
Xа), R = OH, R1 = H
Xb), R = H, R1 =
Oꢀ ꢀDꢀGal
β
Synthesis of cationic glycosurfactants.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 5
2010

SYNTHESIS OF POSITIVELY CHARGED GALACTOSEꢀBEARING SURFACTANTS
659
The starting compound was 6ꢀaminohexanol (I), respectively. Their structures were confirmed by physꢀ
whose amino group served as a “procationic” center of icochemical methods.
the future positively charged lipid, whereas its
To summarize, we proposed a preparative method
for the synthesis of carbohydrateꢀbearing cationic surꢀ
factants based on the Fukuyama reaction with the goal
of studying their antimicrobial activity and their
potential application for the preparation of transfectꢀ
hydroxyl group was used for the addition of carbohyꢀ
drate residues. The reaction of 6ꢀaminohexanol (
with an excess of 2ꢀnitrobenzenesulfonyl chloride in
the presence of triethylamine resulted in sulfonyl
amide (II). Alkylation under the conditions of the
Fukuyama reaction with tetradecyl bromide in the
presence of potassium carbonate led to sulfonyl amide
I)
ing agents. Although the total yield of compounds (Xa
)
and (Xb) was 10–13%, it can be increased by the optiꢀ
mization of separate stages. This approach can be used
for the development of cationic amphiphils with a
more complex structure, for example, comprising digꢀ
lyceride and sterol residues.
(
III) in a yield of 87%.
The key stage of the synthesis was the glycosylation
of compound (III) with 2,3,4,6ꢀtetraꢀ
galactosylꢀ (IVa) or 2,2',3,3',4’,6,6'ꢀheptaꢀ
ꢀlactosyl bromide (IVb) according to the modified
O
ꢀacetylꢀ
α
ꢀDꢀ
Oꢀacetylꢀ
αꢀD
EXPERIMENTAL
Koenigs–Knorr method in a Soxhlet apparatus [33]
with cadmium carbonate as a promoter [33]. As glycꢀ
osyl donors, we used very accessible and highly reacꢀ
tive acetyl bromosugars. The analysis of the reaction
mixture by TLC demonstrated the formation of a
complex mixture, which agreed with that earlier
described in [34]. Along with the major glycosylation
Distilled solvents of domestic production were
used. Tetradecyl bromide, 6ꢀaminohexanol, thiopheꢀ
nol, and DMF were purchased from Merck; 2ꢀ
nitrobenzenesulfonyl chloride was from Aldrich. TLC
was carried out on Silicagel 60 F254 plates (Merck) and
HP TLC RPꢀ18 F254S plates (Merck). The compounds
were visualized with a Ce(SO4)2– phosphomolybdic
acid solution followed by heating to 100°C [35]; the
Dragendorff’s reagent [36] or UV irradiation (254 nm)
were used. For TLC, the following systems were used:
1.5 : 1 toluene–ethyl acetate (A); 2.2 : 1 petroleum
ether–ethyl acetate (B); 2.5 : 1 benzene–ethyl acetate
(C); 1 : 3 toluene–acetone (D); 2: 10 : 1.5 toluene–
acetone–water (E); 10 : 1 methanol–water (F*); and
50 : 1 methanol–25% ammonia (G*) (* for reverseꢀ
phase silica gel). Column chromatography was perꢀ
products,
mixture contained
hydroxyglycosides, and acetate (VII).
β
ꢀglycosides (Va ) and (Vb), the reaction
α
ꢀanomers (VIa) and (VIb), 2ꢀ
For the minimization of the losses of the ꢀglycoꢀ
β
sides, the deacetylation of 2ꢀhydroxyglycosides was
performed in the reaction mixture. After column
chromatography, 1,2ꢀtransꢀglycosides (Va ) and (Vb
)
were obtained in 62 and 49% yields, respectively. Their
1
structures were confirmed by H NMR spectroscopy.
For example, the resonance of the anomeric proton of
β
ꢀglycoside (Va ) at 4.38 ppm and the coupling conꢀ
stant of 8.1 Hz implied the ꢀconfiguration of the glyꢀ
coside bond. In the 1H NMR spectrum of
ꢀglycoside
Vb), resonances of protons at the anomeric centers
were observed at 4.37 and 4.42 ppm with the coupling
constants 1, 2 of 8.1 and 7.8 Hz, respectively. ꢀAnoꢀ
formed on Kieselgel 60 (40–63
μ
m, Merck) and
®
1
β
LiChroprep RPꢀ18 (Merck). H NMR spectra were
registered on a pulse Fourier Bruker MSLꢀ200 or
Bruker MSLꢀ400 spectrometers in CDCl3 or a
CDCl3–CD3OD mixture with Me4Si as the internal
β
(
J
α
standard. Chemical shifts ( ) are given in ppm; J, in
δ
mers of galactoside (VIa) and lactoside (VIb) were isoꢀ
lated in 6% and 12% yields respectively.
Hz. Mass spectra were registered on a timeꢀofꢀflight
Bruker Ultraflex mass spectrometer (Germany) using
the method of laser–desorption ionization. Melting
points were measured on a Boetius microtable (Gerꢀ
many). Optical rotation was measured on a photoelecꢀ
tric Digytor Jasco, model DIP 360, spectropolarimeꢀ
At the final stage, compounds (Va ) and (Vb) were
ꢀdesulfonylated and deacetylated and the cationic
N
head was completed. At first, acetyl groups were
removed because the preserving of the UVꢀdetected
group in the lipid molecule facilitated the purification
of intermediate compounds. Deacetylation with
sodium methylate in methanol resulted in 64 and 68%
of products (VIIIa) and (VIIIb), respectively. The subꢀ
sequent deprotection of the amino group with
thiophenol in the presence of potassium carbonate
gave 76 and 68% of secondary amines (IXa) and (IXb),
respectively. Compounds (IXa) and (IXb) were puriꢀ
fied by column reverseꢀphase chromatography and
their structures were characterized by NMR and mass
spectrometry. The cationic head was completed by the
treatment of secondary amines (IXa) and (IXb) with
methyl iodide in methylethylketone at 70°C. Cationic
ter (Japan). 2,3,4,6ꢀTetraꢀ
bromide (IVa) or 2,2',3,3',4',6,6'ꢀheptaꢀ
ꢀlactosyl bromide (IVb) were obtained as described
in [37, 38].
ꢀ(6ꢀHydroxyhexyl)ꢀ2ꢀnitrobenzenesulfonyl amide
(II). 2ꢀNitrobenzenesulfonyl chloride (2.65 g,
11.96 mmol) was added to a solution of 6ꢀaminohexanol
) (1.17 g, 9.98 mmol), anhydrous triethylamine
Oꢀacetylꢀ
α
ꢀDꢀgalactosyl
Oꢀacetylꢀ
α
ꢀ
D
N
(
I
(3.5 ml), and molecular sieves of 4 Å in anhydrous
dichloromethane (20 ml) cooled to 0°C and the mixꢀ
ture was stirred for 40 h at 20°C. Molecular sieves were
®
filtered off via Celite 545 and washed with dichloꢀ
romethane. The solvent was evaporated in a vacuum
surfactants (Xa) and (Xb) were isolated by column and the residue was chromatographed on silica gel
chromatography on silica gel in 64 and 67% yields, eluted with 100 : 1 chloroform–methanol to give
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 5
2010

660
MOROZOVA et al.
2.41 g (80%) of compound (II); mp 77–78
°
C, Gal), 4.06 (1 H, dd,
J
6.9, 11.2, Haꢀ6 Gal), 4.15 (1 H,
6.5, 11.2, Hbꢀ6 Gal), 4.38 (1 H, d, 8.1, Hꢀ1
Gal), 4.95 (1 H, dd, 3.4, 10.3, Hꢀ3 Gal), 5.13 (1 H, dd,
8.1, 10.3, Hꢀ2 Gal), 5.31 (1 H, dd, 3.4, 0.9, Hꢀ4 Gal),
7.53–7.64 (3 H, m), and 7.92–7.96 (1 H, m, Ar).
ꢀ[6ꢀ(2,3,4,6ꢀTetraꢀ ꢀacetylꢀ ꢀgalactopyraꢀ
nosyloxy)hexyl]ꢀ
Rf 0.38 (A). Found, %: C 47.74; H 6.18; N 9.17.
C12Н18N2O5S. Calc., %: C 47.67; H 6.00; N 9.27. 1H
NMR (200 MHz): 1.25–1.65 (8 H, m,
dd,
J
J
J
J
J
HNCH2(CH2)4CH2OH), 3.05–3.10 (2 H, t,
CH2NH), 3.61 (2 H, t, 6.4,CH2OH), 5.26–5.41
(1 H, m, NH), 7.71–7.80 (2 H, m), 7.83–7.90 (1 H,
m), and 8.08–8.18 (1 H, m, Ar).
Nꢀ(6ꢀHydroxyhexyl)ꢀ ꢀtetradecylꢀ2ꢀnitrobenzeꢀ
nesulfonyl amide (III). Tetradecyl bromide (0.91 ml,
3.06 mmol) was added to a mixture of compound (II
(0.5 g, 1.65 mmol) and potassium carbonate (0.57 g,
4.12 mmol) in DMF (10 ml) at 60 with intense stirꢀ
ring. In 2.5 h, the reaction mixture was diluted with
water (30 ml) and extracted with dichloromethane
3 Na2SO4 20 ml). The extract was washed with water
J 6.4,
J
N
O
α
ꢀD
N
ꢀtetradecylꢀ2ꢀnitrobenzenesulfonyl
amide (VIa), 39.5 mg (6%). Found, %: C 57.53; H
7.85; N 3.27; S 4.01. C40H64N2O14S. Calc., %: C
N
1
57.95; H 7.78; N 3.38; S 3.87. H NMR (400 MHz):
0.82 (3 H, t, J 6.8, (CH2)13CH3), 1.13–1.30 (26 H, m,
)
(CH2)11, (CH2)2), 1.39–1.56 (6 H, m, OCH2CH2,
2 NCH2CH2), 1.93 (3 H, s), 1.98 (3 H, s), 2.01 (3 H,
s) and 2.08 (3 H, s, 4 OCOCH3), 3.20 (4 H, dt, 7.5,
2 CH2N), 3.32 (1 H, dt, 6.4, 9.7, OCHHa), 3.59
(1 H, dt, 76.4, 9.7, OCHHb), 4.00–4.04 (2 H, m, Hꢀ6
Gal), 4.14 (1 H, ddd, 1.3, 5.9, 7.3, Hꢀ5 Gal), 5.01–
5.08 (2 H, m, Hꢀ1, Hꢀ2 Gal), 5.27 (1 H, dd, 3.3,
10.3, Hꢀ3 Gal), 5.38 (1 H, dd, 3.3, 1.3, Hꢀ4 Gal),
7.53–7.65 (3 H, m), and 7.92–7.99 (1H, m, Ar).
ꢀ(6ꢀAcetyloxyhexyl)ꢀ ꢀtetradecylꢀ2ꢀnitrobenzeꢀ
nesulfonyl amide (VII), 69.0 mg (16%); H NMR
(200 MHz): 0.87 (3 H, t, 6.8, (CH2)13CH3), 1.15–
°С
J
J
(
×
(20 ml), dried with Na2SO4, filtered, and evaporated.
The residue was chromatographed on silica gel eluted
with a 5 : 1 petroleum ether–ethyl acetate mixture to
give compound (III) (0.72 g, 87%); Rf 0.61 (B).
Found, %: C 92.85; H 9.19; N 5.52; S 6.09.
J
J
J
N
N
C26H46N2O5S. Calc., %: C 92.62; H 9.30; N 5.62;
1
1
S 6.43. H NMR (200 MHz): 0.88 (3 H, t,
J 6.8,
J
(CH2)13CH3), 1.17–1.38 (26 H, m, (CH2)2, (CH2)11),
1.47–1.66 (6 H, m, OCH2H2, 2 NCH2CH2), 3.22–
3.33 (4 H, m, CH2NCH2), 3.62 (2 H, t, 6.8, CH2OH),
7.58–7.73 (3 H, m) and 7.97–8.05 (1 H, m, Ar).
ꢀ[6ꢀ(2,3,4,6ꢀTetraꢀ ꢀacetylꢀ ꢀgalactopyraꢀ
nosyloxy)hexyl]ꢀ ꢀtetradecylꢀ2ꢀnitrobenzenesulfonyl
1.35 (26 H, m, (CH2)2, (CH2)11), 1.42–1.65 (6 H,
mm, OCH2CH2, 2 NCH2CH2), 2.02 (3 H, s,
COCH3), 3.18–3.31 (4 H, m, 2 CH2N), 4.00 (2 H, t,
J
J
6.5, CH2OH), 7.55–7.71 (3 H, m), and 7.94–8.03
(1 H, m, Ar).
N
O
β
ꢀD
N
N
ꢀ[6ꢀ(2,2',3,3',4',6,6,ꢀHeptaꢀ
O
ꢀacetylꢀ
β
ꢀlactoꢀ
amide (Va). A mixture of compound (III) (0.4 g,
0.8 mmol) and calcined cadmium carbonate (0.28 g,
1.6 mmol) in anhydrous toluene (50 ml) was refluxed
in a Soxhlet apparatus in the presence of calcined
granulated silica gel. After 3 toluene passes via silica
syloxy)hexyl]ꢀ
Nꢀtetradecylꢀ2ꢀnitrobenzenesulfonyl
amide (Vb) was obtained as described for amide (Va
)
from compound (III) (0.43 g, 0.86 mmol), cadmium
carbonate (0.3 g, 1.73 mmol), and 2,2',3,3',4',6,6'ꢀ
heptaꢀ ꢀacetylꢀ ꢀlactosyl bromide (IVb) (1.21 g,
O
α
ꢀD
gel, 2,3,4,6ꢀtetraꢀ
O
ꢀacetylꢀ ꢀDꢀgalactosyl bromide
α
1.73 mmol). The following compounds were isolated
by column chromatography on silica gel (3 : 1 toluꢀ
ene–ethyl acetate): diamide (Vb) (0.47 g, 49%) as a
20
(IVa) (0.66 g, 1.6 mmol) was added in portions for
30 min and the mixture was refluxed for 3.5 h. The
®
reaction mixture was filtered through Celite 545 and
yellowish oil;
–2.5 (c 1, CHC13), Rf 0.47 (A).
the solvent was removed in a vacuum. The residue was
dried in the vacuum of an oil pump and diluted in
anhydrous pyridine (2 ml). Acetic anhydride (0.5 ml)
was added, and the mixture was kept for 4 h at 20–
23°C and evaporated in a vacuum with toluene. The
residue was diluted in chloroform, washed with 3%
[α]D
Found, %: C 55.60; H 7.25; N 2.40. C52H80N2O22S.
Calc., %: C 55.90; H 7.22; N 2.51. H NMR
(400 MHz): 0.81 (3H, t,
1.25 (26 H, m, (CH2)11, (CH2)2), 1.38–1.50 (6 H, m,
OCH2CH2, 2 NCH2CH2), 1.89 (3 H, s), 1.96 (3 H, s),
1.98 (6 H, s), 2.00 (3 H, s), 2.05 (3 H, s), 2.09 (3 H, s,
7 OCOCH3), 3.15–3.22 (4 H, m, 2 CH2N), 3.36 (1 H,
1
J 6.8, (CH2)13CH3), 1.11–
HCl (3
×
5 ml) and water to pH 7.0, and evaporated.
The residue was chromatographed on silica gel eluted
with an 8 : 1 toluene–ethyl acetate mixture to give
dt, J 6.5, 9.7, OCHHa), 3.50–3.56 (1 H, m, Hꢀ5 Lac),
20
0.41 g of compound (Va ) (62%) as yellow oil;
[α]D
3.69–3.76 (2 H, m, OCHHb, Hꢀ4 Lac), 3.78–3.83
(1 H, m, Hꢀ5' Lac), 3.97–4.10 (3 H, m, Haꢀ6, Ha,bꢀ6'
⎯
1.5 (c 1, CHC13), Rf 0.6 (C). Found, %: C 57.41; H
7.80; N 3.29; S 3.99. C40H64N2O14S. Calc., %: C
Lac), 4.37 (1 H, d,
Hꢀ1' Lac), 4.40–4.44 (1 H, m, Hbꢀ6 Lac), 4.81 (1 H,
dd, 8.1, 9.3, Hꢀ2 Lac), 4.88 (1 H, dd, 3.4, 10.3, Hꢀ
3' Lac), 5.04 (1 H, dd, 7.8, 10.3, Hꢀ2' Lac), 5.12
(1 H, t, 9.3, Hꢀ3 Lac), 5.27 (1 H, d, 3.4, Hꢀ4' Lac),
7.52–7.63 (3 H, m), and 7.91–7.96 (1 H, m, Ar).
ꢀ[6ꢀ(2,2',3,3',4',6,6'ꢀHeptaꢀ ꢀacetylꢀ ꢀlactoꢀ
syloxy)hexyl]ꢀ ꢀtetradecylꢀ2ꢀnitrobenzenesulfonyl
amide (VIb), yield 115.0 mg (12%); H NMR
J 8.1, Hꢀ1 Lac), 4.42 (1 H, d, J 7.8,
57.95; H 7.78; N 3.38; S 3.87. Mass, m/z: 851.538
[
M
+ Na]+, 867.529 [
M
+ Na]+; 1H NMR
J 6.8, (CH2)13CH3), 1.12–
J
J
(400 MHz): 0.81 (3H, t,
J
1.28 (26 H, m, (CH2)11, (CH2)2), 1.37–1.53 (6 H, m,
OCH2CH2, 2 NCH2CH2), 1.92 (3 H, s), 1.98 (6 H, s)
J
J
and 2.08 (3 H, s, 4 OCOCH3), 3.19 (4 H, dt,
2 CH2N), 3.38 (1 H, dt, 6.9, 9.7, OCHHa), 3.79
(1 H, dt, 6.2, 9.7, OCHHb), 3.81–3.86 (1 H, m, Hꢀ5
J 7.2, 8.1,
N
O
β
J
N
1
J
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 5
2010

SYNTHESIS OF POSITIVELY CHARGED GALACTOSEꢀBEARING SURFACTANTS
661
(400 MHz): 0.81 (3H,t,
J
6.8, (CH2)13CH3), 1.12–
6ꢀ(
N
ꢀTetradecylamino)hexylꢀ ꢀDꢀgalactopyranoꢀ
β
1.31 (26 H, m, (CH2)11, (CH2)2), 1.39–1.56 (6 H, m, side (IXa). Potassium carbonate (0.25 g, 1.8 mmol)
OCH2CH2, 2 NCH2CH2), 1.91 (3 H, s), 1.97 (3 H, s), and thiophenol (0.18 ml, 1.8 mmol) were added to a
2.00 (3 H, s), 2.04 (3 H, s), 2.08 (3 H, s), 2.09 (3 H, s), solution of compound (VIIIa) (0.12 g, 0.18 mmol) in
2.10 (3 H, s, 7 OCOCH3), 3.14–3.25 (4 H, m, 2 DMF (10 ml) under stirring. In 2 h, the mixture was
®
CH2N), 3.38 (1 H, dt,
(1 H, m, Hꢀ5 Lac), 3.63 (1 H, dt,
3.71–3.65 (1 H, m, Hꢀ4 Lac), 3.82–4.30 (6 H, m, Hꢀ
5, Hꢀ5', Hꢀ6, Hꢀ6' Lac), 4.52 (1 H, d, 7.8, Hꢀ1' Lac),
4.79 (1 H, dd, 4.1, 10.5, Hꢀ2 Lac), 4.89–4.98 (2 H,
m, Hꢀ1, Hꢀ3' Lac), 5.13 (1 H, dd, 7.9, 10.5, Hꢀ2'
Lac), 5.31 (1 H, dd, 3.4, 1.1, Hꢀ4' Lac), 5.30 (1 H, t,
10.5, Hꢀ3 Lac), 7.51–7.65 (3 H, m), and 7.90–7.98
(1 H, m, Ar);
J
6.5, 9.7, OCHHa), 3.50–3.56
filtered through Celite 545 and the solvent was evapꢀ
orated in a vacuum. The residue was chromatographed
on a LiChroprep RPꢀ18 column eluted with a 5 : 1
methanol–5% ammonia mixture to give 0.066 g (77%)
J
6.5, 9.7, OCHHb),
®
J
J
20
of compound (IXa),
+12.7° (
c
1, 1 : 1 CHC13–
[α]D
J
+
CH3OH), Rf 0.5 (D). Mass, m/z: 475.769 [
513.817 [
6.8, (CH2)13CH3), 1.15–1.35 (26 H, m, (CH2)11,
M]
J
+ K]+; 1H NMR (400 MHz): 0.80 (3 H, t,
J
M
J
(CH2)2), 1.39–1.50 (4 H, m, 2 NCH2CH2), 1.51–1.60
(2 H, m, OCH2CH2), 2.52–2.58 (4 H, m, 2 NCH2),
3.07–3.19 (3 H, m, Hꢀ2, Hꢀ3, Hꢀ5 Gal), 3.42 (1 H,
N
ꢀ(6ꢀAcetoxyhexyl)ꢀ
nesulfonyl amide (VII), yield 83.6 mg (18%). 1H NMR
(200 MHz): 0.87 (3 H, t, 6.8, (CH2)13CH3), 1.15–
1.35 (26 H, m, (CH2)2, (CH2)11), 1.42–1.65 (6 H, m,
OCH2CH2, 2 NCH2CH2), 2.02 (3 H, s, COCH3),
3.18–3.31 (4 H, m, 2 CH2N), 4.00 (2 H, t, 6.5,
CH2OH), 7.55–7.71 (3 H, m) and 7.94–8.03 (1 H,
m, Ar).
Nꢀtetradecylꢀ2ꢀnitrobenzeꢀ
J
dt,
3.76 (1 H, m, Hꢀ4 Gal), 3.80 (1 H, dt,
OCHHb), 4.20(1 H, d, 7.8, Hꢀ1 Gal).
6ꢀ( ꢀTetradecylamino)hexylꢀ ꢀlactopyranoside (IXb)
J
6.5, 9.7, OCHHa), 3.67 (2 H, d,
J
6.2, 2 Hꢀ6 Gal),
J
6.5, 9.7,
J
J
N
β
was obtained similarly to compound (IXa) from sulfoꢀ
nyl amide (VIIIb) (0.22 g 0.267 mmol), potassium carꢀ
bonate (0.41 g, 3.0 mmol), and thiophenol (0.4 ml, 3.0
mmol). Compound (IXb) (0.115 g, 68%) was isolated
by chromatography on a LiChroprep RPꢀ18 column
eluted with 5 : 100 : 1 chloroform–methanol–5%
20
Nꢀ[6ꢀ ꢀDꢀgalactopyranosyloxy)hexyl]ꢀNꢀtetradeꢀ
β
cylꢀ2ꢀnitrobenzenesulfonyl amide (VIIIa). A freshly
prepared solution of 1N sodium methylate (1 ml) was
added to a solution of compound (Va ) (0.41 g,
0.49 mmol) in methanol (4 ml) and the mixture was
kept for 30 min at 23°C. The reaction mixture was
neutralized with cationꢀexchange resin Amberlite IRꢀ
120 (H+), filtered, and evaporated. The residue was
chromatographed on a silica gel column eluted with a
ammonia,
–1.2° (
c
1, CH3OH), Rf 0.45 (G).
[α]D
Mass,
m/
z
: 676.524 [M
+ K]+; 1H NMR (400 MHz):
0.80 (3 H, t, J 6.9, (CH2)13CH3), 1.17–1.35 (26 H, m,
(CH2)11, (CH2)2), 1.36–1.58 (6 H, m, 2 NCH2CH2,
OCH2CH2), 2.42–2.48 (4 H, m, 2 NCH2), 3.14–3.82
1 : 1 toluene–acetone mixture to give 0.208 g of comꢀ
20
[α]D
pound (VIIIa) (64%) as a yellow oil;
–1.0° (
c
1,
+ Na]+,
+ K]+; 1H NMR (400 MHz): 0.81 (3 H, t,
6.8, (CH2)13CH3), 1.10–1.29 (26 H, m, (CH2)11,
(14 H, m, Lac protons, OCH2), 4.19 (1 H, d,
Hꢀ1 Lac), 4.29 (1 H, d, 7.5, Hꢀ1' Lac).
ꢀ[6ꢀ ꢀgalactopyranosyloxy)hexyl]ꢀN,Nꢀdiꢀ
methylꢀ ꢀtetradecylammonium iodide (Xa). Cesium
J 7.8,
CHC13), Rf 0.4 (E). Mass, m/z: 683.450 [
699.424 [
M
J
M
J
N
β
ꢀD
(CH2)2), 1.36–1.66 (6 H, m, OCH2CH2,
2 NCH2CH2), 3.15–3.24 (H, m, 2CH2N), 3.40–3.62
(4 H, m, Hꢀ2, Hꢀ3, Hꢀ5 Gal, OCHHa), 3.74–
3.90 (3 H, m, 2 Hꢀ6 Gal, OCHHb), 3.98–4.04 (1 H,
m, Hꢀ4 Gal), 4.17 (1 H, d,
(3 H, m), and 7.90–7.95 (1 H, m, Ar).
N
carbonate (0.055 g, 0.17 mmol) and CH3I (1 ml) were
added to a solution of compound (IXa) (0.046 g,
0.097 mmol) in methylethylketone (1.5 ml) and the
mixture was stirred for 3 h at 70°C. The reaction mixꢀ
ture was filtered off, the solvent was evaporated in a
vacuum, and the residue was chromatographed on a
J 7.2, Hꢀ1 Gal), 7.53–7.65
Nꢀ[6ꢀ ꢀlactosyloxy)hexyl]ꢀ ꢀtetradecylꢀ2ꢀnitrobenꢀ
β
N
®
LiChroprep RPꢀ18 column eluted with 5 : 1 methaꢀ
nol–5% ammonia to give 0.039 g (64%) of compound
zenesulfonyl amide (VIIIb) was obtained similarly to
compound (VIIIa) from (Vb) (0.18 g, 0.16 mmol) and
1 N sodium methylate in methanol (1 ml). Compound
20
(
Ха),
+14.2° (
c
1, CH3OH), Rf 0.8 (D). Mass,
– I]+. 1H NMR (400 MHz): 0.82
6.8, (CH2)13CH3), 1.19–1.48 (26 H, m,
[α]D
(
VIIIb) (0.09 g, 68%) was isolated by chromatography
on LiChroprep RPꢀ18 eluted with a 10 : 1 methanol–
m
/z
: 504.416 [
M
®
(3 H, t,
J
20
[α]D
water mixture.
–1.5° (
c
1, CHC13), Rf 0.45 (F).
Found, %: C 54.58; H 8.31; N 3.07. C38H66N2O15S
1/2 H20. Calc., %: C 54.86; H 8.12; N 3.37. 1H NMR
(400 MHz): 0.80 (3 H, t, 6.7, (CH2)13CH3), 1.12–
(CH2)11, (CH2)2), 1.55–1.68 (m, 2 H, OCH2CH2),
1.69–1.74 (m, 4 H, 2 NCH2CH2), 3.01 (6 H, s,
N(CH3)2), 3.20–3.59 (8 H, m, 2 N+CH2, Hꢀ2, Hꢀ3,
Hꢀ5 Gal, OCHHa), 3.65–3.69 (2 H, m, 2 Hꢀ6 Gal),
×
J
3.78 (1 H, m, Hꢀ4 Gal), 3.84 (1 H, dt,
OCHHb), 4.15 (1 H, d, 7.2, Hꢀ1 Gal).
J 6.5, 9.9,
1.29 (26 H, m, (CH2)11, (CH2)2), 1.38–1.57 (6 H, m,
OCH2CH2, 2 NCH2CH2), 3.15–3.23 (4 H, m,
2 CH2N), 3.24–3.85 (14 H, m, Lac protons, OCH2),
J
N
ꢀ[6ꢀ ꢀLactopyranosyloxy)hexyl]ꢀN,Nꢀdimethylꢀ
ꢀtetradecylammonium iodide (Xb) was obtained simꢀ
Lac), 7.53–7.67 (3 H, m) and 7.89–7.93 (1 H, m, Ar). ilarly to compound (Xa) from lactoside (IXb) (0.059 g,
β
4.21 (1 H, d,
J
7.8, Hꢀ1 Lac), 4.29 (1 H, d,
J
7.5, Hꢀ1'
N
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 5
2010

662
MOROZOVA et al.
17. Gaucheron, J., Wong, T., Wong, K.F., Maurer, N., and
Cullis, P.R., Bioconjugate Chem., 2002, vol. 13, pp.
671–675.
0.092 mmol), cesium carbonate (0.163 g, 0.50 mmol),
and CH3I (1 ml). Compound (Xb) (0.049 g, 67%) was
®
isolated by chromatography on a LiChroprep RPꢀ18
18. Cui, Z. and Mumper, R.J., Bioconjugate Chem., 2002,
column eluted with a 5 : : 1 methanol–water mixture;
1
Rf 0. 5 (E). Mass, m/z: 666.536 [
M
– I]+. H NMR
vol. 13, pp. 1319–1327.
19. Shi, N., Zhang, Y., Zhu, C., Boado, R.J., and Parꢀ
dridge, W.M., Proc. Natl. Acad. Sci. USA, 2001, vol. 98,
pp. 12754–12759.
20. Choosakoonkriang, S., Wiethoff, C.M., Anchordoquy, T.J.,
Koe, G.S., Smith, J.G., and Middaugh, C.R., J. Biol.
Chem., 2001, vol. 276, pp. 8037–8043.
21. Lobo, B.A., Davis, A., Koe, G., Smith, J.G., and Midꢀ
daugh, C.R., Arch. Biochem. Biophys., 2001, vol. 386,
pp. 95–105.
(400 MHz): 0.81 (3 H, t,
J
6.9, (CH2)13CH3), 1.18–
1.39 (26 H, m, (CH2)11, (CH2)2), 1.49–1.79 (6 H, m,
2 NCH2CH2, OCH2CH2), 3.05 (6 H, s, N(CH3)2),
3.10–3.87 (18 H, m, Lac protons, OCH22,
2 N+CH2), 4.15 (1 H, d,
J 7.8, Hꢀ1 Lac), 4.27 (1 H, d,
J
7.5, Hꢀ1' Lac).
ACKNOWLEDGMENTS
22. Kikuchi, I.S. and CarmonaꢀRibeiro, A.M., J. Phys.
Chem. B, 2000, vol. 104, pp. 2829–2835.
The work was supported by the Russian Foundaꢀ
tion for Basic Research, project no. 07ꢀ03ꢀ00632a,
and federal program “Scientific and Teaching Personꢀ
nel of Innovative Russia” (contract no. P715).
23. Banerjee, R., Das P.K., Srilakshmi G.V., Chaudhuri A.,
Rao N.M, J. Med. Chem., 1999, vol. 42, pp. 4292–4299.
24. Banerjee, R., Mahidhar, Y.V., Chaudhuri, A., Gopal, V.,
and Rao, N.M., J. Med. Chem., 2001, vol. 44, pp. 4176–
4185.
REFERENCES
25. Zhang, H., Ma, Y., and Sun, X.ꢀL., Med. Res. Rev.
[Online early access] DOI: 10.1002/med.20171. Pubꢀ
lished Online: July 22, 2009.
1. Merianos, J.J., in Disinfection, Sterilization, and Preserꢀ
vation, Block, S.S., Ed., Lea & Febiger, 1922, pp. 225–
255.
26. Murao, A., Nishikawa, M., Managit, C., Wong, J.,
Kawakami, S., Yamashita, F., and Hashida, M., Pharm.
Res., 2002, vol. 19, pp. 1808–1814.
27. Mukthavaram, R., Marepally, S., Venkata, M.Y.,
Vegi, G.N., Sistla, R., and Chaudhuri, A., Biomateꢀ
rials, 2009, vol. 30, pp. 2369–2384.
2. Cabral, J.P.S., Can. J. Microbiol., 1992, vol. 38,
pp. 115–123.
3. Vieira, D.B. and CarmonaꢀRibeiro, A.M., J. Antimiꢀ
crob. Chemother., 2006, vol. 58, pp. 760–767.
4. Viscardi, G., Quagliotto, P., Barolo, C., Savarino, P.,
Barni, E., and Fisicaro, E., J. Org. Chem., 2000, vol. 65,
pp. 8197ꢀ8203.
28. Alshoaibi, Z.Ya., Morozova, N.G., and Serebrennikꢀ
ova, G.A., Bioorg. Khim., 2000, vol. 26, pp. 703–706
5. Bhattacharya, S. and Mandal, S.S., Biochemistry (US)
1998, vol. 37, pp. 7764–7777.
,
[
Russ. J. Bioorg. Chem. (Eng. Transl.), 2000, vol. 26,
pp. 633–636].
6. Li, S. and Thacker, J., Biochem. Biophys. Res. Comꢀ
mun., 1997, vol. 231, pp. 531–534.
7. Martin, B., Sainlos, M., Aissaoui, A., Oudrhiri, N.,
Hauchecorne, M., Vigneron, J.ꢀP., Lehn, J.ꢀM., and
Lehn, P., Curr. Pharm. Design, 2005, vol. 11, pp. 375–
394.
8. Pinnaduwage, P., Schmitt, L., and Huang, L., Biochim.
Biophys. Acta, 1989, vol. 985, pp. 33–37.
9. Rose, J.K., Buonocore, L., and Whitt, M.A., BioTechꢀ
niques, 1991, vol. 10, pp. 520–525.
29. Alshoaibi, Z.Ya., Andryushina, T.Yu., Morozova, N.G.,
and Serebrennikova, G.A., Bioorg. Khim., 2003, vol. 29,
pp. 323–325 [Russ. J. Bioorg. Chem. (Eng. Transl.), 2003,
vol. 29, pp. 293–295].
30. Quagliotto, P., Viscardi, G., Barolo, C., D’Angelo, D.,
Barni, E., Compari, C., Duce, E., and Fisicaro, E.,
J. Org. Chem., 2005, vol. 70, pp. 9857–9866.
31. Fukuyama, T., Jow, C.ꢀK., and Cheung, M., Tetraheꢀ
dron Lett., 1995, vol. 36, pp. 6373–6374.
32. Fukuyama, T., Cheung, M., Jow, C.ꢀK., Hidai, Y., and
10. Ballas, N., Zakai, N., Sela, I., and Loyter, A., Biochim.
Biophys. Acta, 1988, vol. 939, pp. 8–18.
Kan, T., Tetrahedron Lett., 1997, vol. 38, pp. 5831–5834.
33. Tolkach, A.M., Polonik, S.G., and Uvarova, N.I.,
USSR Inventor’s Certificate no. 1428755, Byull. Izoꢀ
bret., 1988, no. 37.
11. Tabatt, K., Sameti, M., Olbrich, C., Muller, R.H., and
Lehr, C.ꢀM., Eur. J. Pharm. Biopharm., 2004, vol. 57,
pp. 155–162.
12. Masotti, A., Mossa, G., Cametti, C., Ortaggi, G.,
Bianco, A., Del Grosso, N., Malizia, D., and Esposito, C.,
Colloid. Surface. B, 2009, vol. 68, pp. 136–144.
13. Lappalainen, K., Pirila, L., Jaaskelainen, I., Syrjanen,
K., and Syrjanen, S., Anticancer Res., 1996, vol. 16,
pp. 2485–2489.
34. Morozova, N.G., Peredkova, E.V., and Serebrennikꢀ
ova, G.A., Bioorg. Khim., 1996, vol. 22, pp. 799–803
[
Russ. J. Bioorg. Chem. (Eng. Transl.), 1996, vol. 22,
pp. 675–678].
35. Kritchevsky, D. and Kirk, M.R., Arch. Biochem. Bioꢀ
phys., 1952, vol. 35, pp. 346–351.
36. Dawson, R., Elliott, D., Elliott, W., and Jones, K.,
Data for Biochemical Research, Oxford: Clarendon,
1986.
14. Wielbo, D., Shi, N., and Sernia, C., Biochem. Biophys.
Res. Commun., 1997, vol. 232, pp. 794–799.
15. Meyer, O., Kirpotin, D., Hong, K., Sternberg, B.,
Park, J.W., Woodle, M.C., and Papahadjopoulos, D.,
J. Biol. Chem., 1998, vol. 273, pp. 15621–15627.
37. Dahmen, J., Frejd, T., Magnusson, G., and Noori, G.,
Carbohyd. Res., 1982, vol. 111, pp. C1–C4.
16. Hu, Q., Shew, C.R., Bally, M.B., and Madden, T.D., 38. Hudson, C.S. and Johnson, J.M., J. Am. Chem. Soc.
Biochim. Biophys. Acta, 2001, vol. 1514, pp. 1–13. 1915, vol. 37, pp. 1270–1275.
,
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 36
No. 5
2010