Efficient synthesis of phospholipids from glycidyl phosphates

Article abstract of DOI:10.1021/jo010734+

New efficient routes to enantiopure phospholipids, starting from (S)-glycidol, are described. Lysophosphatidic acids and phosphatidic acids were obtained in good overall yields from (S)-glycidol, in only three and four steps, respectively. Moreover, the strategy can also be used to produce phosphatidylcholines in three steps. Using dialkylphosphoramidites, (S)-glycidol was phosphorylated to give (R)-1-O-glycidyl dialkyl phosphates. Regiospecific epoxide opening, using hexadecanol or cesium palmitate, followed by phosphate deprotection, provided lysophosphatidic acids. 2-O-Esterification prior to phosphate deprotection provided 1,2-O-diacyl and 1-O-alkyl-2-O-acyl phosphatidic acids. Phosphorylation of (S)-glycidol using phosphorus oxychloride followed by in situ treatment with choline tosylate produced (R)-glycidyl phosphocholine. Subsequent nucleophilic opening of the epoxide using cesium palmitate produced 1-O palmitoyl-sn-glycero-3-phosphocholine, which has been used in syntheses of phosphatidylcholines.

Full text of DOI:10.1021/jo010734+

194
J . Org. Chem. 2002, 67, 194-199
Efficien t Syn th esis of P h osp h olip id s fr om Glycid yl P h osp h a tes
J an Lindberg, J ohan Ekeroth, and Peter Konradsson*
Department of Chemistry, Linko¨ping University, SE-581 83 Linko¨ping, Sweden
petko@ifm.liu.se
Received J uly 23, 2001
New efficient routes to enantiopure phospholipids, starting from (S)-glycidol, are described.
Lysophosphatidic acids and phosphatidic acids were obtained in good overall yields from (S)-glycidol,
in only three and four steps, respectively. Moreover, the strategy can also be used to produce
phosphatidylcholines in three steps. Using dialkylphosphoramidites, (S)-glycidol was phosphorylated
to give (R)-1-O-glycidyl dialkyl phosphates. Regiospecific epoxide opening, using hexadecanol or
cesium palmitate, followed by phosphate deprotection, provided lysophosphatidic acids. 2-O-
Esterification prior to phosphate deprotection provided 1,2-O-diacyl and 1-O-alkyl-2-O-acyl phos-
phatidic acids. Phosphorylation of (S)-glycidol using phosphorus oxychloride followed by in situ
treatment with choline tosylate produced (R)-glycidyl phosphocholine. Subsequent nucleophilic
opening of the epoxide using cesium palmitate produced 1-O-palmitoyl-sn-glycero-3-phosphocholine,
which has been used in syntheses of phosphatidylcholines.
In tr od u ction
optically active phosphatidylcholine, PAF, and analogues
thereof, starting from racemic glycidol and epichlorohy-
drin,13 the commercially available (S)-glycidol, or (R)-
glycidyl tosylate have been published.14-17 Glycidol de-
rivatives, in the form of glycidyl glycosides, have also
been used in syntheses of glycosyl glycerolipids.18-20 The
methods involving epoxide opening of enantiopure gly-
cidols have been shown to proceed regiospecifically, and
without significant loss of optical purity. Up to date,
however, these methods involve protecting group ma-
nipulations after epoxide opening.
Surprisingly, direct phosphorylation of glycidol and
subsequent opening of the epoxide, has not been pub-
lished. This approach provides an efficient route, without
tedious protecting group manipulations, to various bio-
logically important phospholipids, and analogues thereof.
Phospholipids, especially phosphatidylcholine and phos-
phatidylethanolamine, are major constituents of cell
membranes. Many phospholipids possess biological
activity,1-4 examples are the hormone platelet activating
factor (PAF) and lysophosphatidic acid (LPA)3-5 (Figure
1). For these lipids, the mechanisms of action are not fully
understood. By modification of different parts of the lipid,
activity varies from one biological process to the other.5-7
In addition to structure-activity studies, recent advances
in the construction of artificial cell membranes with
specific biological functions demand tailor-made glycerol-
derived lipids.8
In this paper we present efficient methods for synthesis
of various phospholipids and analogues thereof, starting
from enantiomerically pure (S)-glycidol. Up to date
several routes to glycerophospholipids have been pre-
sented,9-12 most of them involving derivatization and
protecting group manipulations of various selectively
protected glycerol derivatives. Methods, for preparing
Resu lts a n d Discu ssion
In the routes leading to phosphatidic acids, phospho-
rylations were performed with two different dialkyl
phosphoramidites, dibenzyl-N,N-diisopropyl, and di-tert-
butyl-N,N-diisopropyl phosphoramidite. The amidites
were synthesized using known methods.21-24 The phos-
phitylation of (S)-glycidol using dibenzyl-N,N-diisopropyl
phosphoramidite and subsequent oxidation using m-
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10.1021/jo010734+ CCC: $22.00 © 2002 American Chemical Society
Published on Web 12/06/2001

Synthesis of Phospholipids from Glycidyl Phosphates
J . Org. Chem., Vol. 67, No. 1, 2002 195
F igu r e 1. Efficient and short synthetic routes from commercially available (S)-glycidol to many bioactive and/or naturally occurring
glycerophospholipids, developed in this work.
Sch em e 1
Sch em e 2
CPBA, produced the phosphorylated glycidol derivative
1 in 89% yield (Scheme 1). Lewis acid-catalyzed epoxide
opening of the corresponding tosylate, using hexadecanol
and BF3, has been shown to proceed regio- and stereospe-
cifically.15 Opening of the epoxide in 1 using similar
conditions proceeded with complete regiospecificity to
produce the dibenzyl-protected lysoalkyl phosphatidic
acid 2 in 67% yield. Deprotection of the phosphate gave
compound 3, in 53% overall yield from (S)-glycidol. DCC-
promoted esterification of 2 with palmitic acid in almost
quantitative yield, and subsequent debenzylation of the
phosphate produced the 1-O-alkyl-2-O-acyl-phosphatidic
acid 5, in 40% overall yield from (S)-glycidol.
Attempts to produce 1-O-acyl-3-O-dibenzylphosphoryl-
sn-glycerols by nucleophilic opening of the epoxide 1
using cesium palmitate produced only decomposed mate-
rial and benzyl palmitate, due to carboxylate attack on
the benzyl groups. At this stage our attention was turned
to other phosphate protecting groups, not so easily
attacked by nucleophiles. The tert-butyl group seemed
like a reasonable choice. Di-tert-butyl-N,N-diisopropyl
phosphoramidite was used for the oxidative phosphoryl-
ation of (S)-glycidol to produce (R)-di-tert-butyl-phospho-
rylglycidol 6 in 74% yield (Scheme 2). Regioselective
opening of the epoxide with cesium palmitate produced
7a in 71% yield along with minor amounts of the
regioisomeric 3-O-di-tert-butylphosphoryl-2-O-palmitoyl-
sn-glycerol.
Esterification of 7a with oleic acid produced the di-tert-
butyl protected mixed 1,2-diacyl phosphatidic acid 8 in
87% yield. Treatment of 7a and 8 with TFA in CH2Cl2
cleanly produced the deprotected lysophosphatidic acid
9 and the mixed 1,2-diacyl phosphatidic acid 10 in
quantitative yields. The overall yields were 53% and 46%,
respectively. Besides from being present in biological
membranes, these types of compounds have been used
in syntheses of phosphatidylcholines and phosphatidyl
ethanolamines, by coupling with various choline- and
ethanolamine derivatives.14,25-28
(22) Tanaka, T.; Tamatsukuri, S.; Ikehara, T. Tetrahedron Lett.
1986, 27, 199.
(23) Ullman, E.; Engels, J . Tetrahedron Lett. 1986, 27, 1023.
(24) Kuo-Long, Y.; Fraser-Reid, B. Tetrahedron Lett. 1988, 29, 979-
982.
To find a shorter route to choline derivatives, (S)-
glycidol was phosphorylated using phosphorus oxychlo-

196 J . Org. Chem., Vol. 67, No. 1, 2002
Lindberg et al.
Sch em e 3
Sch em e 4
ride. Subsequent in situ treatment with choline tosylate
produced the choline derivative 11 in 53% yield. Nucleo-
philic opening of the epoxide using cesium palmitate
produced lysophosphatidylcholine 12 (Scheme 3), which
has been used in syntheses of phosphatidylcholine.13,29,30
Con clu sion s
Both natural and unnatural enantiomers of a broad
range of biologically important phospholipids, and ana-
logues thereof, can be obtained in high optical purity from
commercially available (S)- or (R)-glycidol. The routes are
short and efficient and proceed in good overall yields.
Phosphorylation of (S)-glycidol using dialkyl phos-
phoramidites, followed by opening of the epoxide, using
alcohol or cesium carboxylate, and subsequent phosphate
deprotection is an effective route to enantiopure lyso-
phosphatidic acids. Moreover, 2-O-esterification of the
lysophosphatidic acids prior to phosphate deprotection,
provides phosphatidic acids in only four steps.
The minor amounts of the regioisomeric 2-O-palmitoyl
derivative, produced in the nucleophilic opening of the
epoxide in Scheme 2, was thought to be the result of acyl
migration from the primary position. There is, of course,
a possibility that part of the product is a result of
nucleophilic attack on the secondary carbon, followed by
equilibration of the regioisomers as depicted in Scheme
4. This would give a partially racemized product. There-
fore the optical purity of 7a had to be evaluated.
Variation of the phosphorylation procedure enables
synthesis of other classes of phospholipids. Phosphory-
lation of (S)-glycidol using phosphorus oxychloride fol-
lowed by in situ treatment with choline tosylate, and
subsequent nucleophilic opening of the epoxide using
cesium palmitate, produced the lysophosphatidylcholine
1-O-Palmitoyl-sn-glycero-3-phosphocholine, which has
been used in syntheses of phosphatidylcholines.13,29,30
7a was converted to Mosher’s acid derivative 13a
(Figure 2) by treatment with (R)-R-methoxy-R-(trifluo-
romethyl)phenylacetic acid chloride ((R)-Mosher’s acid
chloride) in pyridine. The enantiomeric 7b was synthe-
sized analogously from (R)-glycidol, and derivatized with
(R)-Mosher’s acid to produce 13b (Figure 2).
The optical purity was determined by 1H and 19F NMR.
The double doublet at 4.38 ppm in 13a (Figure 2B)
shifted to 4.48 in 13b (Figure 2D) and the trifluoromethyl
peak from 104.44 to 104.36 (Figures 2A and 2C). Only
traces of the double doublet at 4.48 ppm in Figure 2B
and the double doublet at 3.38 in Figure 2D could be
detected, indicating that no loss of optical purity had
occurred. The 19F spectra (Figures 2A and 2C) indicated
>96% ee. Considering the optical purity of the starting
compouds, (S)-glycidol (98% ee) and (R)-Mosher’s acid
chloride (99% ee), no significant loss of optical purity is
observed.
Exp er im en ta l Section
Gen er a l Meth od s. Organic phases were dried over MgSO4,
filtered, and concentrated in vacuo below 40 °C. TLC: 0.25
mm precoated silica gel plates (MERCK silica gel 60F254);
Detection by spraying the plates with AMC solution [(NH4)2-
MoO4 100 g and Ce(IV)SO4 2 g dissolved in 10% H2SO4, 2 L]
followed by heating at ∼250 °C. Optical rotations were
recorded at room temperature with
a Perkin-Elmer 241
polarimeter. Melting points were recorded with a Gallenkamp
melting point apparatus. Flash chromatography (FC): Silica
1
gel MERCK 60 (0.040-0.063 mm). H and 13C NMR spectra
were recorded on a Varian Mercury 300 at 300 MHz (1H), 75
MHz (13C), 121 MHz (31P) and 282 MHz (19F), temp 25 °C.
Chemical shifts are given in ppm with TMS as internal
standard (δ ) 0.00); 31P, 85% H3PO4 (δ ) 0.00); 19F, CFCl3 (δ
) 0.00). MALDI-TOF mass-spectra were recorded with a
Voyager-DE STR Biospectrometry Workstation, in positive
mode, using an R-cyano-4-hydroxycinnamic acid matrix. (S)-
glycidol (98% ee) and (R)-glycidol (98% ee) were purchased
from Aldrich. (R)-R-methoxy-R-(trifluoromethyl)phenylacetic
acid chloride (99% ee) was purchased from Fluka.
(25) Aneja, R.; Chadha, J . S.; Davies, A. P. Tetrahedron Lett. 1969,
48, 4183-4186.
(26) Rakhit, S.; Bagli, J . F.; Deghenghi, R. Can. J . Chem. 1969, 47,
2906-2910.
(27) Bersch, B.; Starck, J . P.; Milon, A.; Nakatani, Y.; Ourisson, G.
Bull. Soc. Chim. Fr. 1993, 130, 575-583.
(28) Akita, H.; Umezawa, I.; Matsukura, H.; Oishi, T. Chem. Pharm.
Bull. 1992, 40, 318-324.
(29) Xia, J .; Hui, Y.-Z. Tetrahedron Asym. 1997, 8, 451-458.
(30) Vodovozova, E. L.; Molotkovsky, J . G. Tetrahedron Lett. 1994,
35, 1933-1936.

Synthesis of Phospholipids from Glycidyl Phosphates
J . Org. Chem., Vol. 67, No. 1, 2002 197
F igu r e 2. 1H and 19F NMR spectra of the (S)-Mosher’s acid derivatives 13a (A+B) and 13b (C+D).
(R)-Diben zylp h osp h or ylglycid ol (1). To a stirred solu-
tion of (S)-glycidol (253 mg, 3.42 mmol) and dibenzyl-N,N-
diisopropyl phosphoramidite (2.50 g, 6.92 mmol) in CH2Cl2
(100 mL) was added 1H-tetrazole (800 mg, 11.4 mmol). After
30 min, the mixture was cooled to 0 °C and m-CPBA (1.75 g,
10 mmol) was added. The mixture was stirred for 40 min,
washed with 10% aqueous Na2S2O3 and NaHCO3 (sat.), dried,
filtered, and concentrated. FC (toluene/EtOAc 3:2, 1% NEt3)
gave 1 as a colorless oil (1.02 g, 3.05 mmol, 89%). Rf 0.38
(toluene/EtOAc 1:1); [R]D -8.9 (c 0.88, CHCl3); 1H and 13C NMR
spectra were identical to those published for the racemic
compound.31
3-O-Diben zylph osph or yl-1-O-h exadecyl-sn -glycer ol (2).
To a solution of 1 (80 mg, 0.24 mmol) and hexadecanol 116
mg, 0.48 mmol) in CH2Cl2 (3 mL) was added a 10% v/v solution
of BF3-diethyl ether complex in CH2Cl2 (40 µL). After 10 h,
the mixture was concentrated. FC (toluene/EtOAc 2:1) of the
residue gave crystalline 2 (93 mg, 0.16 mmol, 67%). Rf 0.43
(toluene/EtOAc 1:1): mp 44-45 °C (from hexane); [R]D -2.7
(c 1.1, CHCl3); NMR (CDCl3): 1H, δ 0.88 (t, 3H, J ) 6.4 Hz),
1.26 (br, 26H), 1.53 (m, 2H), 3.37-3.42 (m, 4H), 3.87-4.13 (m,
3H), 5.05 (d, 2H, J ) 8.4 Hz), 5.05 (d, 2H, J ) 8.4 Hz), 7.34
(br, 10H); 13C, δ 14.1, 22.7, 26.1, 29.3, 29.5, 29.6, 29.7, 31.9,
69.1 (d, J ) 5.5 Hz), 69.3 (d, J ) 7.4 Hz), 70.7, 71.7, 128.0,
128.6, 135.7 (d, J ) 5.5 Hz); 31P, δ 0.58; Anal. Calcd for
C
33H53O6P: C 68.7; H 9.3 Found C 68.8; H 8.9.
1-O-Hexa d ecyl-3-O-p h osp h or yl-sn -glycer ol (3). A mix-
ture of 2 (26 mg, 0.045 mmol) and 10% Pd on carbon (18 mg)
in EtOAc (2 mL) was stirred under H2 (1 atm) for 2 h, filtered
through Celite, and concentrated to yield crystalline 3 (16 mg,
0.040 mmol, 90%). Rf 0.12 (CHCl3/MeOH/H2O 65:25:4); mp.
66-67 °C (CHCl3); [R]D -2.0 (c 1.1, DMSO); 1H and 13C NMR
spectra were identical to those published for the racemic
compound.32
(31) De Viguerie, N. L.; Willson, M.; Pe´rie´, J . New J . Chem. 1994,
18, 1183-1195.
(32) Malekin, S. I.; Khromova, N. Y.; Kisin, A. V.; Kruglyak, Y. L.;
Kurochkin, V. K. Russ. J . Bioorg. Chem. 1997, 23, 677-682.

198 J . Org. Chem., Vol. 67, No. 1, 2002
Lindberg et al.
3-O-Diben zylp h osp h or yl-1-O-h exa d ecyl-2-O-p a lm itoyl-
sn -glycer ol (4). A solution of 2 (40 mg, 0.069 mmol), palmitic
acid (28 mg, 0.11 mmol), dicyclohexylcarbodiimide (23 mg, 0.11
mmol), and 4-(dimethylamino)pyridine (1 mg, 0.008 mmol) in
CH2Cl2 (2 mL) was stirred for 22 h, filtered through Celite,
and concentrated. FC (toluene/EtOAc 8:1) of the residue gave
crystalline 4 (54 mg, 0.066 mmol, 96%). Rf 0.27 (toluene/EtOAc
8:1); mp 37-38 °C (from ethanol); [R]D -3.1 (c 1.0, CHCl3);
NMR (CDCl3): 1H, δ 0.88 (m, 6H), 1.25 (br, 52H), 1.45-1.62
(m, 4H), 2.27, (t, 2H, J ) 7.7 Hz), 3.35-3.41 (m, 2H), 3.49 (d,
2H, J ) 5.5 Hz), 4.08-4.23 (m, 2H), 5.03 (d, 2H, J ) 8.0 Hz),
5.04 (d, 2H, J ) 8.0 Hz), 5.11 (m, 1H), 7.34 (br, 10H); 13C, δ
14.1, 22.7, 24.9, 26.1, 29.1, 29.3, 29.4, 29.7, 31.9, 34.3, 66.1 (d,
J ) 5.5 Hz), 68.3, 69.3, 69.4. 70.6 (d, J ) 7.4 Hz), 71.8, 127.9,
128.5, 128.6, 135.8 (d, J ) 7.4 Hz), 173.1; 31P, δ 0.53. Anal.
Calcd for C49H83O7P: C 72.2; H 10.3. Found: C 72.2; H 10.1.
1-O-Hexa d ecyl-2-O-p a lm itoyl-3-O-p h osp h or yl-sn -glyc-
er ol (5). A mixture of 4 (33 mg, 0.040 mmol) and 10% Pd on
carbon (14 mg) in EtOAc (2 mL) was stirred under H2 (1 atm)
for 2 h, filtered through Celite, and concentrated to yield
crystalline 5 (20 mg, 0.032 mmol, 78%). Rf 0.36 (CHCl3/MeOH/
H2O 65:25:4); mp 78-80 °C (from EtOAc/hexane); [R]D -1.3
172.9, 173.3; 31P, δ -9.02. Anal. Calcd for C45H87O8P: C 68.7;
H 11.1. Found: C 68.3; H 11.0.
1-O-P a lm itoyl-3-O-p h osp h or yl-sn -glycer ol (9). To 7a (37
mg, 0.071 mmol) in CH2Cl2 (8 mL) was added TFA (2 mL).
After 10 min was added MeOH (0.10 mL). After an additional
10 min, the mixture was diluted with toluene (20 mL) and
concentrated to yield crystalline 9 (29 mg, 0.071 mmol, quant.).
Rf 0.12 (CHCl3/MeOH/H2O 65:25:4); mp 79-80 °C (from
CHCl3); [R]D +5.0 (c 0.54, DMSO), lit.35 (sodium salt, CHCl3)
[R]D +13.2; NMR (DMSO): 1H, δ 0.84 (t, 3H, J ) 5.8 Hz), 1.22
(br, 24H), 1.50 (m, 2H), 2.28 (t, 2H, J ) 7.4 Hz), 3.72-3.85
(m, 3H), 3.93 (dd, 1H, J ) 11.2, 5.6 Hz), 4.02 (dd, 1H, J )
11.2, 4.1 Hz); 13C, δ 13.8, 22.0, 24.3, 28.4, 28.59, 28.61, 28.8,
28.89, 28.94, 31.2, 33.3, 64.7, 66.1 (d, J ) 5.4 Hz), 67.2 (d, J )
8.0 Hz), 172.7; 31P, δ 0.25; MALDI-TOF Calcd for C19H39O7P:
[M + Na]+ 433.2. Found: [M + Na]+ 433.2.
2-O-Oleoyl-1-O-p a lm itoyl-3-O-p h osp h or yl-sn -glycer ol
(10). To 8 (32 mg, 0.041 mmol) in CH2Cl2 (8 mL) was added
TFA (2 mL). After 10 min was added MeOH (0.1 mL). After
an additional 10 min, the mixture was diluted with toluene
(20 mL) and concentrated to yield 10 (28 mg, 0.041 mmol,
quant.). Rf 0.64 (CHCl3/MeOH/H2O 65:25:4); mp diammonium
salt 185-187 °C (from CHCl3/Acetone), lit.36 192-194 °C; [R]D
+7.2 (c 3.0, CHCl3), lit.36 [R]D +4.5; NMR (CDCl3/CD3OD 4:1):
1H, δ 0.81-0.91 (m, 6H), 1.2-1.4 (m, 48H), 1.55-1.68 (m, 4H),
1.97-2.06 (m, 4H), 2.30-2.37 (m, 6H), 4.08-4.12 (m, 2H), 4.19
(dd, 1H, J ) 12.1, 6.3 Hz), 4.38 19 (dd, 1H, J ) 12.0, 3.7 Hz),
5.20-5.26 (m, 1H), 5.32-5.37 (m, 2H); 13C, δ 14.2, 22.9, 25.1,
27.4, 29.3-30.0, 32.1, 34.3, 34.4, 62.4, 64.5 (d, J ) 5.2 Hz),
70.1 (d, J ) 7.4 Hz), 129.9, 130.2, 173.7, 174.2; 31P (CDCl3), δ
1.62.
(R)-Glycid yl P h osp h och olin e (11). To a stirred solution
of (S)-glycidol (670 mg, 9.04 mmol) and ethyl N,N-diisopropy-
lamine (6.5 mL, 9.5 mmol) in CHCl3 (20 mL) at 0 °C under an
argon atmosphere was added phosphorus oxychloride (1.40 g,
9.13 mmol). After 2 h, pyridine (2.0 mL) and choline tosylate
(2.89 g, 10.5 mmol) were added, and the mixture was allowed
to attain room temperature. After an additional 5 h, water was
added (0.50 mL), and the stirring was continued for an
additional 1 h. Concentration and FC (70% EtOH) gave 11
(1.14 g, 4.76 mmol, 53%). Rf 0.19 (70% EtOH); [R]D -15 (c 1.8,
MeOH); NMR (CD3OD): 1H, δ 2.68 (dd, 1H, J ) 5.0, 2.7 Hz),
2.82 (dd, 1H, J ) 5.0, 4.1 Hz), 3.21-3.26 (m, 1H), 3.24 (s, 9H),
3.65-3.70 (m, 3H), 4.19 (ddd, 1H, J ) 12.0, 6.9, 2.5 Hz), 4.26-
4.33 (m, 2H); 13C, δ 45.0, 52.0 (d, J ) 8.3 Hz), 54.6, 54.7, 54.8,
60.4 (d, J ) 5.2 Hz), 67.5, 67.7 (d, J ) 5.4 Hz); 31P, δ 0.47;
MALDI-TOF Calcd for C8H19NO5P: [M + H]+ 240.1. Found:
[M + H]+ 240.1.
1-O-P alm itoyl-sn -glycer o-3-ph osph och olin e (12). A mix-
ture of 11 (107 mg, 0.45 mmol), cesium palmitate (174 mg,
0.45 mmol) and palmitic acid (115 mg, 0.55 mmol) in DMF (5
mL) was stirred at 80 °C. After 26 h, the mixture was
concentrated. FC (MeOH/H2O 5:1) gave crystalline 12 (143 mg,
0.29 mmol, 65%). Rf 0.29 (MeOH/H2O 5:1); mp 250-253 °C
(from MeOH), lit.37 243-245 °C; [R]D -2.8 (c 7.7, CHCl3/MeOH
9:1), lit. for the enantiomer 3-O-palmitoyl-sn-glycero-1-phos-
phocholine.37 [R]D +2.8; 1H and 13C NMR spectra were identical
to those previously published.38 31P (CD3OD), δ 1.22.
1
(c 1.7, CHCl3); H and 13C NMR of the pyridinium salt were
in accordance with those published for the racemic com-
pound.33
(R)-Di-ter t-bu tylp h osp h or ylglycid ol (6). To a stirred
solution of (S)-glycidol (238 mg, 3.21 mmol) and di-tert-butyl-
N,N-diisopropyl phosphoramidite (1.58 g, 5.70 mmol) in CH2-
Cl2 (100 mL) was added 1H-tetrazole (760 mg, 10.9 mmol).
After 30 min, the mixture was cooled to 0 °C and m-CPBA
(1.70 g, 9.85 mmol) was added. The mixture was stirred for
40 min, washed with 10% aqueous Na2S2O3 and NaHCO3
(sat.), dried, filtered, and concentrated. FC (gradient petroleum
ether 65-75/EtOAc 3:1 f 1:1, 0.1% NEt3) gave 6 as a colorless
oil (635 mg, 2.38 mmol, 74%). Rf (toluene/EtOAc 1:1) 0.24; [R]D
1
-5.0 (c 1.1, CHCl3); H NMR (CCl4) were in accordance with
that published for the racemic compound.34 NMR (CDCl3): 1H,
δ 1.52 (s, 18H), 2.68 (dd, 1H, J ) 4.7, 2.6 Hz), 2.84 (dd, 1H, J
) 4.6, 4.6 Hz), 3.25 (m, 1H), 3.91 (ddd, 1H, J ) 11.7, 8.0, 5.8
Hz), 4.19 (ddd, 1H, J ) 11.7, 3.3, 6.9); 13C, δ 29.9 (d, J ) 3.7
Hz), 44.7, 50.2 (d, J ) 9.2 Hz), 67.4 (d, J ) 5.6 Hz), 82.8 (d, J
) 3.8 Hz), 83.0 (d, J ) 3.7); 31P, δ -9.09.
3-O-Di-t er t -b u t ylp h osp h or yl-1-O-p a lm it oyl-sn -gly-
cer ol (7a ). A mixture of 6 (127 mg, 0.48 mmol), cesium
palmitate (557 mg, 1.43 mmol), and palmitic acid (122 mg, 0.48
mmol) in DMF (5 mL) was stirred at 80 °C. After 10 h, the
mixture was allowed to attain room temperature, and the
precipitate filtered off. Concentration and FC (toluene/EtOAc
2:1 1% NEt3) gave 7a as a waxy solid (178 mg, 0.34 mmol,
71%). Rf 0.52 (toluene/EtOAc 1:3); [R]D -1.7 (c 1.5, CHCl3);
NMR (CDCl3): 1H, δ 0.88 (t, 3H, J ) 6.6 Hz), 1.26 (br, 26H),
1.50 (s, 18H), 1.62 (m, 2H), 2.34 (t, 2H, J ) 7.5 Hz), 3.98-
4.17 (m, 5H); 13C, δ 14.1, 22.7, 24.9, 29.2, 29.3, 29.4, 29.5, 29.6,
29.7, 29.8, 29.9, 31.9, 34.2, 64.5, 68.2 (d, J ) 7.4 Hz), 68.9 (d,
J ) 5.5 Hz), 83.2 (d, J ) 7.4 Hz), 173.8; 31P, δ -7.32. Anal.
Calcd for C27H55O7P: C 62.0; H 10.6. Found: C 61.9; H 10.4.
3-O-Di-ter t-bu tylp h osp h or yl-2-O-oleoyl-1-O-p a lm itoyl-
sn -glycer ol (8). A solution of 7a (51 mg, 0.098 mmol), oleic
acid (74 mg, 0.26 mmol), dicyclohexylcarbodiimide (54 mg,
0.262 mmol), and 4-(dimethylamino)pyridine (2 mg, 0.016
mmol) in CH2Cl2 (3 mL) was stirred for 20 h, filtered through
Celite, and concentrated. FC (toluene/EtOAc 8:1, 0.1% NEt3)
gave 8 (67 mg, 0.085 mmol, 87%) as a colorless syrup. Rf 0.32
(toluene/EtOAc 4:1); [R]D +4.9 (c 0.9, CHCl3); NMR (CDCl3):
1H, δ 0.88 (m, 6H), 1.26 (br, 48H), 1.48 (s, 18H), 1.61 (m, 4H),
2.01 (m, 4H), 2.31 (t, 2H, J ) 7.5 Hz), 2.32 (t, 2H, J ) 7.5 Hz),
4.05-4.10 (m, 2H), 4.17 (dd, 1H, J ) 11.9, 6.0 Hz), 4.36 (dd,
1H, J ) 11.9, 4.2 Hz), 5.18-5.41 (m, 3H); 13C, δ 14.1, 22.7,
24.9, 27.2, 27.3, 29.1, 29.2, 29.3, 29.5, 29.6, 29.7, 29.8, 29.9,
31.9, 34.1, 34.2, 62.0, 64.4 (d, J ) 5.5 Hz), 69.6 (d, J ) 9.2
Hz), 82.7 (d, J ) 7.4 Hz), 82.8 (d, J ) 7.4 Hz), 129.7, 130.0,
3-O-Di-ter t-bu tylp h osp h or yl-2-O-[(S)-R-m eth oxy-R-(tr i-
flu or om et h yl)p h en yla cet yl]-1-O-p a lm it oyl-sn -glycer ol
(13a ). A solution of 7a (17 mg, 0.033 mmol) and (R)-R-methoxy-
R-(trifluoromethyl)phenylacetic acid chloride (32 mg, 0.13
mmol) in pyridine (2 mL) was stirred for 21 h. The mixture
was diluted with CH2Cl2, washed with aqueous NaHCO3,
dried, filtered, and concentrated. FC (toluene f toluene/EtOAc
2:1 0.1% NEt3) gave 13a as a white solid (23 mg, 0.31 mmol,
(35) Stoffel, W.; Wolf, G. D. Hoppe-Seyler’s Z. Physiol. Chem. 1966,
347, 94-101.
(36) Molotkowsky, J . G.; Nikulina, L. F.; Bergelson, L. D. Chem.
Phys. Lipids 1976, 17, 108-110.
(37) Eibl, H.; Westphal, O. Liebings Ann. Chem. 1970, 738, 161-
(33) Malekin, S. I.; Kruglyak, Y. L.; Khromova, N. Y.; Aksenova,
M. Y.; Sokolov, V. P. Russ. J . Bioorg. Chem. 1997, 23, 591-596.
(34) Kluba, M.; Zwierak, A. Roczniki Chem. 1974, 48, 1603-1608.
169.
(38) Noda, N.; Tanaka, R.; Nishi, M.; Inoue, S.; Miyahara, K. Chem.
Pharm. Bull. 1993, 41, 1366-1368.

Synthesis of Phospholipids from Glycidyl Phosphates
J . Org. Chem., Vol. 67, No. 1, 2002 199
96%). Rf 0.44 (toluene/EtOAc 2:1); NMR: 1H (CDCl3), δ 0.88
(t, 3H, J ) 6.6 Hz), 1.26 (br, 24H), 1.46 (br, 20H), 2.22 (t, 2H,
J ) 8.1 Hz), 3.58 (s, 3H), 4.05-4.24 (m, 3H), 4.38 (dd, 1H, J )
4.0, 12.4 Hz), 5.51 (m, 1H), 7.40 (m, 3H), 7.54 (m, 2H); 19F
(CCl4), δ 104.36.
1-O-Di-ter t-bu tylp h osp h or yl-2-O-[(S)-R-m eth oxy-R-(tr i-
flu or om et h yl)p h en yla cet yl]-3-O-p a lm it oyl-sn -glycer ol
(13b). 13b was synthesized by the same method as 13a ,
starting from (R)-glycidol. Rf 0.44 (toluene/EtOAc 2:1); NMR:
1H (CDCl3), δ 0.88 (t, 3H, J ) 7.0 Hz), 1.25 (br, 24H), 1.44 (br,
20H), 2.28 (t, 2H, J ) 7.7 Hz), 3.57 (s, 3H), 4.0-4.25 (m, 3H),
4.48 (dd, 1H, J ) 3.3, 12.1 Hz), 5.51 (m, 1H), 7.41 (m, 3H),
7.55 (m, 2H); 19F (CCl4), δ 104.44.
J O010734+