Synthesis of amphiphilic glycophospholipids based on β-cyclodextrins

Article abstract of DOI:10.1007/s11172-009-0033-3

An approach to the synthesis of amphiphilic glycophospholipids based on β-cyclodextrin (β-CD) was proposed. Distinctive features of the proposed method are the use of highly efficient trivalent phosphorus derivatives in the first steps of the synthesis an

Full text of DOI:10.1007/s11172-009-0033-3

Russian Chemical Bulletin, International Edition, Vol. 58, No. 1, pp. 223—229, January, 2009
223
Synthesis of amphiphilic glycophospholipids based on βꢀcyclodextrins*
M. K. Grachev,a S. V. Sipin,a L. O. Kononov,b and E. E. Nifant´eva
aMoscow Pedagogical State University,
3 Nesvizhskii per., 119021 Moscow, Russian Federation.
Fax: +7 (495) 246 7766. Eꢀmail: chemdept@mtuꢀnet.ru, phosphorus@rambler.ru
bN. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences,
47 Leninsky prosp., 119991 Moscow, Russian Federation.
Fax: +7 (499) 135 5328. Eꢀmail: kononov@ioc.ac.ru
An approach to the synthesis of amphiphilic glycophospholipids based on βꢀcyclodextrin
(βꢀCD) was proposed. Distinctive features of the proposed method are the use of highly
efficient trivalent phosphorus derivatives in the first steps of the synthesis and the regioselective
monophosphorylation of nonprotected βꢀCD. The efficiency of using 13С NMR spectroscopy
for the determination of the position and number of phospholipid residues introduced into the
CD composition was shown.
Key words: βꢀcyclodextrin, phospholipidocyclodextrins, regioselected monophosphorylation,
NMR spectroscopy.
It is known that βꢀcyclodextrin (βꢀCD) and its nuꢀ
merous derivatives found wide use in pharmacology mainly
as drug “containers” due to their unique ability to “enꢀ
capsulate” various hydrophobic compounds (formation
of host—guest inclusion compounds).1,2 This encapsulaꢀ
tion protects the included drug from biodegradation, inꢀ
creases its solubility, and, which is especially important,
favors the efficient and selective delivery of the drug to
the required site within the predetermined period. Attenꢀ
tion has recently been given to studies of covalent “linkꢀ
ing” (conjugation) of drugs to CD, which in several cases
makes it possible to develop drugs of more prolonged and
purposeful effect.3—5
In addition to the extending role of drug mediators,
CD are used for the extraction of bioregulators (first of
all, cholesterol) and study of their metabolism in live
biological systems.2 It was found6 that hydroxypropylated
βꢀCD can extract virionꢀbound** cholesterol and affect
cholesterol bound to HIVꢀinfected cells. This extraction
of cholesterol strongly decreases the formation of viruses,
and the respective virions released from the cells that have
been liberated from cholesterol were minimally virulent.
In addition, CD can in vivo affect the transfer of cell
signals through the biological membrane6,7 changing the
composition of the soꢀcalled lipid rafts.8
The progress in the considered directions could be the
combination of the said specific properties of CD with
the transport properties of organized structures, such as
vesicles and micelles, or the introduction of CD derivaꢀ
tives into preꢀorganized lipid matrices, for example, lipoꢀ
somes or their analogs (see, e.g., Ref. 9). First representaꢀ
tives of a new class of amphiphilic CDꢀbased derivatives,
namely, βꢀCDꢀbound cholesterol (A) and phospholipidoꢀ
cyclodextrins (B), have recently been synthesized aimed
at design of fragments close in structure and nature of the
lipid cholesterol matrix localized in the cell membranes.10—13
These compounds contain hydrophobic lipid fragments
conjugated with the narrow part of the CD rim, which
imparts the amphiphilic properties and remains a wide
part of the CD rim free for “guest” inclusion. Cholesterol
was chosen as a hydrophobic residue because of its presꢀ
ence in biological membranes, due to which studies of its
role and influence on the behavior of bilayers are of
special interest.14 The second representative of amphiphilic
CD (B) contains the phospholipid residue attached to the
CD framework through the spacer. It is considered that
the control of the length of this spacer can optimize the
incorporation of the molecule into the lipid matrix to
form the soꢀcalled “membrane anchor” and thus induce
lesser structural changes in the matrix.10—12 It is of interest
that the methylated analogs of the phospholipid systems
(R = Me) are more soluble in water than the nonprotected
ones (R = H), which is important for the formation of
selfꢀorganized objects (vesicles and micelles). The low
values of critical micelle concentrations suggest strong
* Dedicated to Academician A. I. Konovalov on his 75th birthday.
** Virion is the completely formed virus particle consisting of
a nucleic acid and a protein shell. Virion stores and transfers the
genetic material from one cell to another.
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 1, pp. 221—227, January, 2009.
1066ꢀ5285/09/5801ꢀ0223 © 2009 Springer Science+Business Media, Inc.

224
Russ.Chem.Bull., Int.Ed., Vol. 58, No. 1, January, 2009
Grachev et al.
R = H, Me
proneness of these amphiphilic molecules to selfꢀorganiꢀ
zation in water. It was shown by static and dynamic light
scattering that in aqueous solutions these amphiphilic CD
behave as long fluctuating “twigs” (“rods”) corresponding
to infinitely long micelles with bulky CD heads rocking at
the sides. The NMR spectra recorded with the known
guest molecules showed that even in the highly organized
form these amphiphilic CD completely retain their ability
to include “guests” into the CD cavity.10—12
In addition, it is necessary to provide for any efficient
drug carrier to be able to penetrate through biological
barriers. Among the latter, the most important is the bloodꢀ
brain barrier (BBB) that protects the central nervous
system from foreign substances circulating in blood and
controls the composition and properties of the spinal fluid.
The effective use of, e.g., a neuroactive drug assumes that
the drug can penetrate this selective barrier. The studies
for the model system showed that some phospholipids
containing the βꢀCD residue can penetrate through this
barrier and, which is especially important, do not decomꢀ
pose it. It is considered that the conjugates mentioned are
the first example of drug mediators capable of penetrating
the BBB without violating its integrity.10—12 Thus, due to
a combination of properties of CD as host molecules with
the transport efficiency of selfꢀorganized amphiphilic
molecules, the conjugates can become valuable objects of
special interest for the point delivery of drugs and studyꢀ
ing their metabolism.
synthesis of amphiphilic CD belonging to the class of
phospholipidocyclodextrins. Amphiphilicity of these comꢀ
pounds should be determined by the presence of hydroꢀ
philic hydroxy groups of CD molecules and the presence
of hydrophobic residues of aliphatic fatty acids, and
the ability to inclusion is determined by the presence of
the CD cavity. What we propose is the use of CD with free
hydroxy groups. Since the stated problem is experimenꢀ
tally difficult, we decided first to select optimal condiꢀ
tions for the synthesis of compounds containing CD linked
through the phosphate group with the 1,2ꢀОꢀisoproꢀ
pylideneglycerol residue (1) modeling natural 1,2ꢀdiglyceꢀ
rides. Phenyl phosphodichloridite (2) was used as the
starting phosphorylating agent (Scheme 1).
The reaction was carried out in diethyl ether at –15 to
–18 °C in the presence of a tenfold molar excess of steriꢀ
cally hindered base, ethyldiisopropylamine (hydrogen
chloride acceptor), to suppress the possible phosphorylaꢀ
tion of the secondary hydroxy groups of CD in the subꢀ
sequent stage of phosphorylation16,17 (method A) or in
pyridine at –5 °C (method B). The reaction mixture
containing chloride 3 was added to a solution of CD in
pyridine (molar ratio 1 : 1) at –15 to –18 °C, and
triphosphite 4a that formed was oxidized by the hydrogen
peroxide—urea adduct (“hydroperit”) to the correspondꢀ
ing phosphate 5a (Scheme 1)*.
Alternatively, chloride 3 was added to a solution of
CD in pyridine at –5 °C, and the resulting triphosphite 4b
Taking into account our data on a possibility of
regioselective acylation15 and monophosphorylation of the
primary hydroxy groups in the presence of the secondary
groups,16 we intended to develop other approaches to the
* For discussion convenience, products 4 and 5 synthesized by
methods A and B (see further) were designated as 4a and 4b, 5a
and 5b, respectively.

Amphiphilic glycophospholipids
Russ.Chem.Bull., Int.Ed., Vol. 58, No. 1, January, 2009
225
Scheme 1
Excess
Method A
Method B
X = O (5a,b), S (6)
i. H2O2•C(O)(NH2)2; ii. H2O2•C(O)(NH2)2; S
was oxidized to the corresponding phosphate 5b or treated
with sulfur to form thiophosphate 6.
no inclusion compounds according to the data from
1Н NMR spectroscopy. An important feature of the oneꢀ
pot syntheses of compounds 5 and 6 according to methꢀ
ods A and B is that all the three steps before the isolation
of the final PV products should be carried out at the indiꢀ
cated reduced temperatures for at most 20—30 min to
avoid the accumulation of considerable amounts of poorly
separable byꢀproducts.
At all steps of the syntheses, the compositions of the
reaction mixtures and the course of the reaction were
monitored by 31Р NMR spectroscopy. Several important
features were observed for phosphorylation in the menꢀ
tioned solvents (among the solvents that are not proton
donors, βꢀcyclodextrin is rather well soluble only in pyriꢀ
dine and DMF). For example, when the reactions are
carried out at ambient temperature, the decomposition
products of PIII derivatives are rapidly accumulated
(mainly due to disproportionation). Under these phosꢀ
phorylation conditions, chloride 3 turned out to be espeꢀ
cially unstable, and triphosphites 4a and 4b, whose conꢀ
tent in the reaction mixture was ~50%, were more stable.
Small distinctions of the 31Р NMR chemical shifts of
compounds 4a and 4b with the same structure should be
specially mentioned: broadened singlets at δ 136.6 (4a)
and 138.0 (4b).* The low total yields of phosphates 5a
and 5b and thiophosphate 6 (12—15% over three steps)
are mainly related to losses upon crystallization from
water (compound 5a) and column chromatography (comꢀ
pounds 5b and 6). According to the data from 13Р NMR
spectroscopy, they appear as singlets at δ 4.6 (5a, 5b) and
53.2 (6). According to the 1Н and 13С NMR data,
ethyldiisopropylamine is included in product 5a. The reꢀ
placement of diethyl ether by toluene as a solvent resulted
in inclusion of toluene in product 5a under the same
conditions of synthesis and isolation. Thus, method B
should be preferred, because products 5b and 6 contained
The structures of isolated compounds 5a, 5b, and 6
1
were confirmed by the data from Н, 13С, and 31Р NMR
1
spectroscopy. The H NMR spectrum of compound 5a
contains additional signals for ethyldiisopropylamine
included into the CD cavity: the broadened signal for
protons of the CH3 groups at δ 1.27 and the multiplet of
protons of the CH2 and CH groups at δ 3.18—3.22. The
signals for ethyldiisopropylamine included in compound
5a also appear in the 13С NMR spectrum as four singlets:
signals for the СH3CH2 and (CH3)2CH groups at δ 14 and
23 and signals for the CH2 and CH groups at δ 44 and 46,
respectively. Note that the signals for ethyldiisopropylꢀ
amine do not disappear after additional recrystallizations
from water and drying. Among other features, we can
mention good resolution of the signals for protons of the
CD framework, which is usually observed for the incluꢀ
sion of an appropriate guest18 into the CD cavity, most
likely, because the CD framework becomes more rigid.
Based on the analysis of the 13С NMR spectra, we can
conclude that the phosphorylation of βꢀCD by phosphoꢀ
dichloridite 3 involved, as expected,15,16 the primary
hydroxy groups. We observed the spinꢀspin coupling
2
constant JP,С(6A)** and the downfield shift of the signals
* Different chemical shifts for compounds of the same structure
are probably caused by the inclusion of Pri NEt into the cavity
** The atom of the glucose CD fragment containing the subꢀ
stituent is marked.
2
of CD 4a (see further).

226
Russ.Chem.Bull., Int.Ed., Vol. 58, No. 1, January, 2009
Grachev et al.
Scheme 2
R = Ph (2, 9, 11), CH2CH2CN (8, 10, 12); X = LEP (9, 10), O (11, 12)
The structures of isolated compounds 11 and 12 were
for the С(6A) atoms in products 5a and 5b (δС 63.4)
compared to the corresponding signal in free βꢀCD
(δС 61.4) and the upfield characteristic shift of the С(5)
signal at δС 73.7 (in free βꢀCD) to δС 71.1 for С(5A).19—22
It is important that no shifts of signals for the secondary
С(2) and С(3) carbon atoms of the CD framework were
observed in the 13С NMR spectra of products 5a and 5b,
i.e., these positions were not affected by phosphorylation.
The individual character of isolated compounds 5a, 5b,
and 6 was confirmed by the TLC data. Compounds 5a
and 5b had the same chromatographic mobilities and
slightly different temperatures of melting onset (with
decomposition): 164 °C (5a) and 167 °C (5b), which is
evidently related to the fact that product 5а is an incluꢀ
1
confirmed by the data from Н, 13С, and 31Р NMR specꢀ
troscopy and MALDIꢀTOF mass spectrometry.
1
Note that the Н NMR spectrum of compound 11
exhibits well resolved signals for protons of the CD frameꢀ
1
work, unlike the Н NMR spectrum of compound 12,
which is probably a consequence of the inclusion of the
phenyl radical into the CD cavity.
Similarly to compounds 5a and 5b, we analyzed the
13С NMR spectra and made a conclusion that the phosꢀ
phorylation of βꢀCD involved the primary hydroxy groups.
We observed the spinꢀspin coupling constant 2JP,С(6A) and
the downfield shift of signals from the С(6A) atoms in
products 11 and 12 (δС 63.1 for C(6A)) compared to the
corresponding signals for C(6) in free βꢀCD (δС 61.4).
The characteristic upfield shift of the signal for С(5) at
δС 73.7 (in free βꢀCD) to δС 71.3 for С(5А) was also
observed.19—22 In addition, in the 13C NMR spectra the
signals for the carbonyl groups of compounds 11 and 12
appear as four signals and the cyano group in compound
12 is observed as two signals suggesting the presence of
diastereomers.
An additional confirmation of the structures of comꢀ
pounds 11 and 12 was obtained by the integration of the
signals for the carbon atoms of these compounds in the
13С NMR spectra recorded with a large delay (RD = 5)
between pulses (Fig. 1).
Thus, we proposed the new routes for the synthesis of
the CD—phospholipid derivatives. The proposed approach
differs from the known method10—12 in the use (at the first
step of the design of complicated systems) of the correꢀ
sponding highly reactive trivalent phosphorus derivatives
instead of the reactant traditional for the chemistry of
natural compounds and in the phosphorylation of βꢀCD
sion compound (5a = 5b ⊃ Pri NEt).
2
Thus, we showed for the first time that highly reactive
phosphochloridites can be used for the regioselective
monophosphorylation of the primary hydroxy groups of
βꢀcyclodextrin in the presence of unprotected secondary
hydroxy groups.
Taking into account the data from model experiments,
we synthesized amphiphilic CD derivatives containing
the covalently linked phospholipid residue using method
B. 1,2ꢀDiꢀОꢀpalmitoylglycerol 7 reacted with freshly disꢀ
tilled phenyl dichloridite 2 or βꢀcyanoethyl phosphoꢀ
dichloridite 8. Treatment of βꢀCD in pyridine at –5 °C
with the chlorides formed and oxidation of βꢀCD triphosꢀ
phites 9 and 10 with hydroperit gave the corresponding
phosphates 11 and 12.
The course of the reaction and compositions of the
reaction mixtures were monitored by 31Р NMR spectroscopy.
Triphosphites 9 and 10 appeared in the 31Р NMR spectra
as broadened singlets at δ 134 and 140, and phosphates 11 and
12 gave broadened singlets at δ 4.2 and 4.5, respectively.

Amphiphilic glycophospholipids
Russ.Chem.Bull., Int.Ed., Vol. 58, No. 1, January, 2009
227
CH2–CH2CH3
O
C–CH2–CH2
CH2–CH3
C(2)
CH3
O
C(3)
C(5) C(6)
O
–O–C–Pal
C≡N
C(4)
C(1)
C–CH2
CN–CH2
δ
160 150 140 130 120 110 100
90
70
60
50
40
30
20
170
80
C(2)
C(5)
C(3)
C(6)
POCH2CHCH2
CN–CH2CH2
C(5A) CH
C(6A)
PO–CH2CH
δ
74
73
72
71
70
69
68
67
65
64
63
62
61
60
59
58
75
66
Fig. 1. 13С NMR spectrum (Pyꢀd5) of compound 12.
containing no protecting groups by the proposed reacꢀ
tants. It can be expected that the new route of synthesis
and the synthesized compounds will find practical use in
studies of functioning cellular membranes and metabolism
of a series of biologically important compounds, as well as
in works devoted to problems of targeted drug delivery.

228
Russ.Chem.Bull., Int.Ed., Vol. 58, No. 1, January, 2009
Grachev et al.
109.9 (1 С, С(СН3)2); 120.4 (2 С, Сoꢀarom); 125.7 (1 С,
С
Experimental
pꢀarom); 130.0 (2 С, Сmꢀarom); 161.7 (1 С, Сipsoꢀarom). 31Р NMR
(Py), δ: 4.6 br.s.
All experiments with trivalent phosphorus compounds were
Method B. 1,2ꢀOꢀIsopropylideneglycerol (1) (0.132 g,
1.00 mmol) in pyridine (3 mL) was added with stirring at –5 °C
over 5 min to a solution of phenyl phosphodichloridite 2 (0.195
g, 1.00 mmol) in pyridine (3 mL). The reaction mixture was
immediately (without isolation of intermediate phosphodichloꢀ
ridite 3) was added with stirring to a solution of βꢀCD (1.00 g,
0.88 mmol) in pyridine (20 mL) at –5 °C over 10 min; the
31Р NMR spectrum of the reaction mixture, δ : 138.0 br.s
(compound 4b). Crushed hydroperit (0.103 g, 1.1Р0 mmol) was
added to the reaction mixture, which was stirred for 24 h at
20 °C. The solvent was distilled off in vacuo. Compound 5b was
purified by column chromatography eluting with system A.
The solvents were removed in vacuo, and the residue was kept
in vacuo (1 Torr) for 5 h at 70 °C. The yield was 0.185 g (15% for
three stages), m.p. (decomp.) 167 °C, Rf 0.74 (A). Found (%):
С, 46.18; Н, 6.06. С54Н85О40Р. Calculated (%): С, 46.16; Н,
carried out in anhydrous solvents purified according to standard
procedures and under an inert atmosphere. 1Н NMR spectra
were recorded on Bruker WMꢀ250 and Bruker АСꢀ200 instruꢀ
ments (250 and 200 MHz, respectively). 13С NMR spectra were
measured on a Bruker АСꢀ200 instrument (50.32 MHz). The
1Н NMR chemical shifts are presented relative to the residual
signal of Py (δ 8.75 for the downfield signal (s)), and the
13С NMR chemical shifts are given relative to the signal for
Pyꢀd5 (δ 149.8 for the downfield signal (t)). 31Р{H} NMR spectra
were recorded on Bruker WPꢀ80SY and Bruker АСꢀ200 instruꢀ
ments (compounds 11 and 12) (32.4 and 82.2 MHz) relative to
85% Н3РО4 as the external standard. Preparative column
chromatography was carried out on Kieselgel 40/60 (Merck),
and TLC analysis was performed on glass plates with silica gel 60
F 254 (Merck), detection of spots in iodine vapor or under
the UV light after spraying with anisaldehyde. The systems for
TLC and column chromatography were acetonitrile—methanol—
water, 3 : 3 : 1 (A) and acetonitrile—chloroform—methanol,
2 : 2 : 1 (B).
Mass spectra were recorded on a Bruker Ultra Flex instruꢀ
ment with the timeꢀofꢀflight (TOF) detector by matrixꢀactivated
laser desorption and ionization (MALDI) (λ = 337 nm) using
2,5ꢀdihydroxybenzoic acid as the matrix.
Commercial βꢀCD (Novodex, Russia) was subjected
to additional thorough dehydration.15 Phenyl phosphodichloꢀ
ridite was obtained by an earlier described procedure.23
1,2ꢀOꢀIsopropylideneglycerol was from Sigma and hydroperit
was from Tatkhimfarmpreparaty (Russia).
1
6.10. The Н, 13C NMR (Pyꢀd5, δ) and 31P NMR spectra are
identical to those of sample 5a. MALDIꢀTOF, m/z: 1405.3 [M]+.
racꢀ6ꢀОꢀ[(2,3ꢀIsopropylidenedioxypropoxy)(phenoxy)thioꢀ
phosphoryl]ꢀβꢀcyclodextrin (6). 1,2ꢀOꢀIsopropylideneglycerol
(1) (0.132 g, 1.00 mmol) in pyridine (3 mL) was added with
stirring to a solution of phenyl phosphodichloridite (2) (0.195 g,
1.00 mmol) in pyridine (3 mL) at –5 °C for 5 min. The reaction
mixture was immediately added with stirring to a solution of
βꢀCD (1.00 g, 0.88 mmol) in pyridine (20 mL) at –5 °C over
10 min; the 31Р NMR spectrum of the reaction mixture;
δ : 138.0 br.s (compound 4b). Finely divided sulfur (0.035 g,
1Р.10 mmol) was added to the reaction mixture, which was stirred
for 24 h at 20 °C. The solvent was distilled off in vacuo. Comꢀ
pound 6 was purified by column chromatography eluting with
system A. The solvents were removed in vacuo, and the residue
was kept in vacuo (1 Torr) for 5 h at 70 °C. The yield was 0.163 g
(13%), Rf 0.71 (A), m.p. (decomp.) 176 °C. Found (%): С, 45.68;
Н, 6.00. С54Н85О39РS. Calculated (%): С, 45.63; Н, 6.03.
1Н NMR (Pyꢀd5), δ: 1.30 (s, 3 Н, CH3); 1.42 (s, 3 Н, CH3); 4.12
(dd, 7 Н, С(2)Н, 3JH(1),H(2) = 2.8 Hz, 3JH(32),H(3) = 9.4 Hz); 4.32
racꢀ6ꢀОꢀ[(2,3ꢀIsopropylidenedioxypropoxy)phenoxy)ꢀ
phosphoryl]ꢀβꢀcyclodextrin (5). Method A. 1,2ꢀOꢀIsopropyliꢀ
deneglycerol (1) (0.123 g, 1 mmol) and ethyldiisopropylamine
(1.423 g, 11.0 mmol) in diethyl ether or toluene (3 mL) were
added to a solution of phenyl phosphodichloridite (2) (0.195 g,
1 mmol) in diethyl ether or toluene (3 mL) at –15 to –18 °C for
5 min. The reaction mixture was immediately (without isolation
of intermediate phosphodichloridite 3) added with stirring to
a solution of βꢀCD (1.00 g, 0.88 mmol) in pyridine (20 mL) at
–15 to –18 °C for 10 min; the 31Р NMR spectrum of the reacꢀ
3
(dd, 7 Н, С(4)Н, JH(3),H(4) = 9.0 Hz, JH(4),H(5) = 9.9 Hz);
4.46—4.55 (m, 21 Н, С(5)Н, С(6)Н2, 3JH(4),H(5) = 9.9 Hz); 4.84
(dd, 7 Н, С(3)Н); 4.91—5.44 (m, 5 Н, СН2—СН—СН2); 5.67
(d, 7 Н, С(1)Н); 7.21—7.48 (m, 5 Н, Нꢀarom). 31Р NMR (Py),
δ: 53 br.s.
tion mixture, δ : 136.6 br.s (compound 4a). Crushed hydroperit
Р
(0.103 g, 1.1 mmol) was added to the reaction mixture, which
was stirred for 24 h at 20 °C. The solvent was distilled off
in vacuo. Compound 5a was purified by recrystallization from
water (2´50 mL). The crystalline product was kept in vacuo
(1 Torr) for 5 h at 70 °C. The yield was 0.148 g (12% over three
racꢀ6ꢀОꢀ[(1,2ꢀDipalmitoyloxypropyloxy)(phenoxy)phosphoꢀ
ryl]ꢀβꢀcyclodextrin (11). 1,2ꢀDiꢀOꢀpalmitoylglycerol23 (7)
(0.50 g, 0.88 mmol) in pyridine (3 mL) was added with stirring
to a solution of phenyl phosphodichloridite 2 (0.195 g, 1.00 mL)
in pyridine (3 mL) at –5 °C for 5 min. The reaction mixture was
immediately added with stirring to a solution of βꢀCD (1.00 g,
0.88 mmol) in pyridine (20 mL) at –5 °C for 10 min; the
31Р NMR spectrum of the reaction mixture, δ : 134.0 br.s (comꢀ
pound 9). Crushed hydroperit (0.103 g, 1.10Рmmol) was added
to the reaction mixture, which was stirred for 24 h at 20 °C. The
solvent was distilled off in vacuo. Compound 11 was purified
by column chromatography eluting with system B. The
solvents were removed in vacuo, and the residue was kept
in vacuo (1 Torr) for 5 h at 70 °C. The yield was 0.194 g (12%),
m.p. (decomp.) 186 °C, Rf 0.57 (B). Found (%): С, 54.20;
Н, 7.76. С83Н141О42Р. Calculated (%): С, 54.12; Н, 7.72.
1Н NMR (Pyꢀd5), δ: 0.87 (m, 3 Н, CH3); 1.03 (m, 3 Н, CH3);
1.29 (m, 48 Н, (СН2)12); 1.68 (m, 4 Н, С(О)СН2СН2); 2.42
1
steps), m.p. (decomp.) 164 °C, Rf 0.74 (А). Н NMR* (Pyꢀd5),
δ: 1.31 (s, 3 Н, CH3); 1.41 (s, 3 Н, CH3); 4.14 (dd, 7 Н, С(2)Н,
3JН(1),Н(2) = 2.9 Hz, 3JН(2),Н(3) = 9.5 Hz); 4.30 (dd, 7 Н, С(4)Н,
3JН(3),Н(4) = 9.1 Hz, 3JН(4),Н(5) = 9.9 Hz); 4.46—4.57 (m, 21 Н,
3
С(5)Н, С(6)Н2, JН(4),Н(5) = 9.9 Hz); 4.82 (dd, 7 Н, С(3)Н);
4.88—5.46 (m, 5 Н, СН2—СН—СН2); 5.68 (d, 7 Н, С(1)Н);
7.20—7.49 (m, 5 Н, Нꢀarom.). 13С NMR* (Pyꢀd5), δ: 26.9
(1 С, СН3); 29.7 (1 С, СН3); 61.4 (6 С, С(6)); 62.1 (1 С,
2
РОСН2СНСН2); 63.4 (С(6А), JС(6),(А)Р = 5.0 Hz); 66.7
2
(1 С, РОСН2СН, JС,Р = 3.6 Hz); 70.1 (1 С, РОСН2СН,
3JС,Р = 6.8 Hz); 71.1 (1 С, С(5А); 73.7 (6 С, С(5)); 74.1 (7 С,
С(3)); 74.5 (7 С, С(2)); 83.2 (7 С, С(4)); 103.8 (7 С, С(1));
* For the signals of ethyldiisopropylamine, see discussion of the
obtained results.

Amphiphilic glycophospholipids
Russ.Chem.Bull., Int.Ed., Vol. 58, No. 1, January, 2009
229
(m, 4 Н, С(О)СН2); 4.13 (dd, 7 Н, С(2)Н, 3JH(1),H(2) = 2.9 Hz,
3JH(2),H(3) = 9.9 Hz); 4.28 (dd, 7 Н, С(4)Н, 3JH(3),H(4) = 8.0 Hz,
3JH(4),H(5) = 9.1 Hz); 4.48 (dd, 7 Н, С(6)Н, 3JH(5),H(6´) = 9.9 Hz,
2JH(6),H(6´) = 14.3 Hz); 4.53 (dd, 7 Н, С(6)Н´, 3JH(5),H(6´) = 9.9 Hz,
2JH(6),H(6´) = 14.3 Hz); 4.67 (m, 7 Н, С(5)Н, 3JH(4),H(5) = 9.1 Hz,
3JH(5),H(6´) = 9.9 Hz, 3JH(5),H(6) = 9.9 Hz); 4.80 (dd, 7 Н, С(3)Н);
4.50—5.10 (m, 5 Н); 5.64 (d, 7 Н, С(1)Н); 7.15 (m, 2 Н, Нmꢀarom);
7.40 (m, 1 Н, Нpꢀarom); 7.52 (d, 2 Н, Ноꢀarom). 13С NMR (Pyꢀd5),
δ: 14.3 (2 С, СН3); 22.8 (2 С, С(О)СН2СН2); 25.2 (2 С,
СН2СН2СН3); 29.1—29.6 (20 С, СН2); 31.9 (2 С, СН2СН3);
34.0, 34.2 (2 С, С(О)СН2СН2); 61.4 (6 С, С(6)); 62.0 (1 С,
3. K. Uekama, K. Minami, F. Hirayama, J. Med. Chem., 1997,
40, 2755; K. Minami, F. Hirayama, K. Uekama, J. Pharm.
Sci., 1998, 87, 715; F. Hirayama, M. Kamada, H. Yano,
K. Udo, H. Arima, K. Uekama, J. Incl. Phenom., 2002,
44, 159; H. Yano, F. Hirayama, M. Kamada, H. Arima,
K. Uekama, J. Cont. Rel., 2002, 79, 103.
4. G. I. Kurochkina, N. A. Kudryavtseva, M. K. Grachev,
S. A. Lysenko, L. K. Vasyanina, E. E. Nifant´ev, Zh.
Obshch. Khim.
Transl.), 2007, 77, 442].
5. G. I. Kurochkina, N. A. Kudryavtseva, M. K. Grachev,
S. A. Lysenko, E. E. Nifant´ev, Zh. Obshch. Khim. 2007,
77, 485 Russ. J. Gen. Chem. (Engl. Transl.), 2007, 77, 450].
, 2007, 77, 477 [Russ. J. Gen. Chem. (Engl.
2
РОСН2СНСН2); 63.3 (С(6А), JС(6А),Р = 4.9 Hz); 66.7 (1 С,
,
РОСН2СН, 2JС,Р = 4.0 Hz); 70.2 (1 С, РОСН2СН, 3JС,Р = 7.1 Hz);
71.4 (1 С, С(5А)); 73.9 (6 С, С(5)); 74.3 (7 С, С(3)); 74.7 (7 С,
С(2)); 83.2 (7 С, С(4)); 103.7 (7 С, С(1)); 119.9 (2 С, Соꢀarom);
125.39 (1 С, Сpꢀarom); 129.78 (2 С, Сmꢀarom); 163.52 (1 С, Сipsoꢀarom).
31Р NMR (Pyꢀd5), δ: 4.2 br.s. MALDIꢀTOF, m/z: 1841.8 [M]+.
racꢀ6ꢀОꢀ[(2,3ꢀDipalmitoyloxypropyloxy)(2ꢀcyanoethoxy)ꢀ
phosphoryl]ꢀβꢀcyclodextrin (12). Dipalmitoylglycerol 7 (0.50 g,
0.88 mmol) in pyridine (3 mL) was added with stirring to a
solution of βꢀcyanoethyl phosphodichloridite24 (8) in pyridine
(3 mL) at –5 °C for 5 min. The reaction mixture was immediꢀ
ately added with stirring to a solution of βꢀCD (1.00 g, 0.88 mmol) in
pyridine (20 mL) at –5 °C over 10 min; the 31Р NMR spectrum
[
6. D. Graham, E. Chertova, J. Hilburn, L. Arthur, J. E. K.
Hildreth, J. Virol., 2003, 77, 8237.
7. Z. Liao, D. Graham, J. E. K. Hildreth, AIDS Res. Human
Retroviruse, 2003, 19, 675.
8. M. Sugahara, M. Uragami, S. L. Regen, J. Am. Chem. Soc.,
2003, 125, 13040.
9. T. Sukegawa, T. Furuike, K. Niikura, A. Yamagishi,
K. Monde, S.ꢀI. Nishimura, Chem. Commun., 2002, 430.
10. S. Moutard, B. Perly, P. Gode, G. Demailly, F. Djedaпniꢀ
Pilard, J. Incl. Phen. Macrocycl. Chem., 2002, 44, 317.
11. B. Perly, S. Moutard, F. DjedaïniꢀPilard, PharmaChem: Fine,
Specialty & Performance Chemicals, 2005, 4, 4.
12. S. Moutard, Relation entre la structure et les propriétés
d´ordanisation de nouvelles cyclodextrines amphiphiles,
Ph. D. Dissertation, Université de Picardie Jules Verne
Picardie, Oct, 2003.
13. A. Janshoff, C. Steinem, A. Michalke, C. Henke, H.ꢀJ. Galla,
Sensors and Actuators B, 2000, 70, 243.
14. R. AuzélyꢀVelty, B. Perly, O. Taché, T. Zemb, P. Jéhan,
P. Guenot, J.ꢀP. Dalbiez, F. DjedaïniꢀPilard, Carbohydr.
Res., 1999, 318, 82.
15. A. E. Glazyrin, A. N. Syrtsev, G. I. Kurochkina, L. O.
Kononov, M. K. Grachev, E. E. Nifant´ev, Izv. Akad. Nauk,
Ser. Khim., 2003, 225 [Russ. Chem. Bull., Int. Ed., 2003, 52,
237].
16. M. K. Grachev, S. V. Sipin, G. I. Kurochkina, E. E.
Nifant´ev, Zh. Obshch. Khim., 2005, 75, 1631 [Russ. J. Gen.
Chem. (Engl. Transl.), 2005, 75, 1556].
17. K. Ishihara, H. Kurihara, H. Yamamoto, J. Org. Chem.,
1993, 58, 3791.
18. A. Douhal, Chem. Rev., 2004, 104, 1955.
of the reaction mixture, δ : 140.0 br.s (compound 10). Crushed
Р
hydroperit (0.103 g, 1.1 mmol) was added to the reaction mixꢀ
ture, which was stirred for 24 h at 20 °C. The solvent was
distilled in vacuo. Compound 12 was purified by column chroꢀ
matography eluting with system B. The solvents were removed
in vacuo, and the residue was kept in vacuo (1 Torr) for 5 h at
70 °C. The yield was 0.176 g (11%), m.p. (decomp.) 196 °C,
Rf 0.55 (B). Found (%): С, 52.78; Н, 7.80. С80Н140NО42Р.
1
Calculated (%): С, 52.83; Н, 7.76. Н NMR (Pyꢀd5), δ: 0.88
(m, 6 Н, CH3); 1.28—1.30 (m, 48 Н, (СН2)12); 1.71 (m, 4 Н,
С(О)СН2СН2); 2.39 (m, 4 Н, С(О)СН2); 3.16 (m, 2 Н, СН2СN);
4.11 (dd, 7 Н, С(2)Н, 3JH(1),H(2) = 3.4 Hz, 3JH(2),H(3) = 9.5 Hz);
3
3
4.22 (t, 7 Н, С(4)Н, JH(3),H(4) = 9.5 Hz, JH(4),H(5) = 9.5 Hz);
4.45—4.56 (m, 23 Н, С(5)Н, С(6)Н2, СН2СН2CN,
3JH(4),H(5) = 9.5 Hz); 4.25—4.90 (m, 5 Н, СН2—СН—СН2);
4.75 (t, 7 Н, С(3)Н); 5.61 (d, 7 Н, С(1)Н). 13С NMR (Pyꢀd5),
δ: 14.3 (2 С, СН3); 22.6 (1 С, СН2СN); 23.0 (2 С, С(О)СН2СН2);
25.2 (2 С, СН2СН2СН3); 29.1—30.0 (20 С, СН2); 32.2 (2 С,
СН2СН3); 34.1, 34.2, 34.3, 34.5 (2 С, С(О)СН2СН2); 61.5 (6 С,
С(6)); 62.1 (1 С, РОСН2СНСН2); 63.1 (1 С, РОСН2СН2СN);
2
63.4 (С(6А), JС(6)(А)Р = 5.0 Hz); 66.7 (1 С, РОСН2СН,
19. D. Rong, V. T. D´Souza, Tetrahedron Lett., 1990, 31, 4275.
20. H.ꢀJ. Schneider, F. Hacket, V. Ruidiger, H. Ikeda, Chem.
Rev., 1998, 98, 1755.
21. P. Forgo, V. T. D´Souza, J. Org. Chem., 1999, 64, 306.
22. P. Forgo, V. T. D´Souza, Tetrahedron Lett., 1999, 40, 8533.
23. E. E. Nifant´ev, A. I. Zavalishina, Khimiya elementoorganiꢀ
cheskikh soedinenii [Chemistry of Organoelement Compounds],
Moscow Pedagogical State University, Moscow, 1980, 91 pp.
(in Russian).
3
2JС,Р = 4.1 Hz); 70.2 (1 С, РОСН2СН, JС,Р = 7.2 Hz); 71.3
(1 С, С(5А)); 73.9 (6 С, С(5)); 74.3 (7 С, С(3)); 74.7 (7 С,
С(2)); 83.4 (7 С, С(4)); 104.0 (7 С, С(1)); 118.0, 119.2 (1 С,
СN); 173.1, 173.2, 173.4, 173.6 (2 С, С(О)). 31Р NMR (Pyꢀd5),
δ: 4.5 br.s. MALDIꢀTOF: m/z, 1818.7 [M]+.
This work was financially supported by the Russian
Foundation for Basic Research (Project No. 08ꢀ03ꢀ00374)
and the Council on Grants at the President of the Russian
Federation (Program of State Support for Leading Scienꢀ
tific Schools of the Russian Federation, Grant 5515.2006.3).
24. P. V. Ioannou, G. H. Dodd, B. T. Golding, Synthesis,
1979, 939.
25. W. Bannwarth, A. Trzeciak, Helv. Chim. Acta, 1987, 70, 175.
References
1. K. Uekama, F. Hirayama, T. Irie, Chem. Rev., 1998, 98, 2045.
2. M. E. Davis, M. E. Brewster, Nature Rev. Drug Discovery,
2004, 3, 1033.
Received December 11, 2007;
in revised form June 3, 2008