Synthesis of glycoglycerolipid of 1,2-dipalmitoyl-3-(N-palmitoyl-6′- amino-6′-deoxy-α-D-glucosyl)-sn-glycerol and its analogues, inhibitors of human Myt1-kinase

Article abstract of DOI:10.1016/j.carres.2012.04.005

A glycoglycerolipid 1a isolated from a marine alga showed inhibition to Myt1 kinase with IC₅₀ of 0.12 μg/mL. We synthesized 1a and its seven analogues (1b-h) in an efficient method with high stereoselectivity. The process employed trichloroacetimidate donor 4b at low substrate concentration to achieve high α-selectivity (α/β = 33:1) in glycosylation reaction. The present synthesis provided various acyl derivatives required for the study on the structure-activity relationship later.

Full text of DOI:10.1016/j.carres.2012.04.005

Carbohydrate Research 355 (2012) 6–12
Contents lists available at SciVerse ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Synthesis of glycoglycerolipid of 1,2-dipalmitoyl-3-(N-palmitoyl-60-amino-60-
deoxy-
a-D-glucosyl)-sn-glycerol and its analogues, inhibitors of human
Myt1-kinase
⇑
Yihua Sun, Jun Zhang, Chunxia Li , Huashi Guan, Guangli Yu
Key Laboratory of Marine Drugs, Ministry of Education of China, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
A glycoglycerolipid 1a isolated from a marine alga showed inhibition to Myt1 kinase with IC50 of 0.12 lg/
Received 20 February 2012
Received in revised form 5 April 2012
Accepted 6 April 2012
mL. We synthesized 1a and its seven analogues (1b–h) in an efficient method with high stereoselectivity.
The process employed trichloroacetimidate donor 4b at low substrate concentration to achieve high
selectivity ( /b = 33:1) in glycosylation reaction. The present synthesis provided various acyl derivatives
required for the study on the structure–activity relationship later.
a
-
a
Available online 26 April 2012
Ó 2012 Elsevier Ltd. All rights reserved.
Keywords:
Glycoglycerolipid
Glycosylation
a-Selectivity
Myt1-kinase inhibitor
1. Introduction
a: R = palmitoyl
b: R = hexanoyl
c: R = octanoyl
d: R = lauroyl
NHR
O
HO
HO
Glycoglycerolipids isolated from marine organisms have re-
cently shown a wide spectrum of biological activities, including
antitumor, anti-HIV-1 infection, and anti-inflammatory activi-
ties.1–4 The 6-deoxy-6-aminoglycoglycerolipids, rarely occurred
as glycoglycerolipids in nature, exhibit unique bioactivities, such
as antibacterial, Myt1 kinase inhibitory, and antileishmanial activ-
ities.5–8 Among the bioactive aminoglycoglycerolipids, glycolipid
1a (Fig. 1), which was isolated from a marine alga with high inhib-
OR
OH
e: R = myristoyl
f: R = stearoyl
O
OR
g: R = isovaleryl
h: R = hydrocinnamoyl
1
Figure 1. Structures of natural aminoglycoglycerolipid 1a and its analogues 1b–h.
studies have shown that the minor structural changes of lipophilic
acyl chain of glycolipids exert positive effect on the antitumor
activity of glycolipids.13 To ascertain the influence of acyl chains
on the inhibitory activity of Myt1-kinase, we synthesized a series
of analogues 1b–h (Fig. 1) of 1a. The structures of compounds
1b–h are different from the natural compound 1a mainly in the
length (b–f), branch (g), and aromaticity (h) of the acyl chains,
which are homogenous at C60, C1, and C2. Although the synthetic
method of compound 1a has been reported,12,14 the procedure
was tedious and the stereoselectivity of glycosylation was still
unsatisfied. Here we report a concise and high stereoselective
method for the synthesis of these compounds for further struc-
ture–activity relationship research.
itory activity on the enzyme Myt1-kinase (IC50 0.12
tracts our great interest.
l
g/mL),6 at-
The human Myt1-kinase is an important regulator of the G2/M
transition in the cell cycle. Myt1 specifically mediates phosphory-
lation at Thr-14 and Tyr-15 of Cdc2-kinase activity and the inhibi-
tion of phosphorylation of Cdc2 is important for timing the entry
into mitosis. Various studies have shown that premature activation
of Cdc2 would lead to mitotic dysfunction and apoptosis.9 Inhibi-
tion of Myt1-kinase is predicted to cause premature activation of
Cdc2.10,11 Therefore inhibitors of Myt1-kinase are supposed to kill
rapidly proliferating cells and interfere with cell cycle checkpoints.
Such inhibitors could represent an extension to conventional che-
motherapy and overcome drug resistance.
With the aim to search for potential Myt1-kinase inhibitors,
2. Results and discussion
glycoglycerolipid 1a has been synthesized by our group.12 Previous
Our synthetic strategy is depicted in Scheme 1. It is noteworthy
that compounds 1a–h have four common structural features,
which contain 60-amino group, homogeneous acyl chains at C60,
⇑
Corresponding author. Tel.: +86 532 82032030; fax: +86 532 82033054.
E-mail address: lchunxia@ouc.edu.cn (C. Li).
0008-6215/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.carres.2012.04.005

Y. Sun et al. / Carbohydrate Research 355 (2012) 6–12
NHR NH2
7
O
O
HO
BnO
BnO
HO
OH
OR
OH
OBn
OH
O
OR
O
2
Target compounds
1a-h
N3
N3
O
O
O
BnO
BnO
BnO
+
HO
O
LG
BnO
O
OBn
OBn
5
O
4
O
3
Scheme 1. Retrosynthetic analysis.
C1, and C2 positions,
a
glycosidic linkage bond and S-chiral center.
was dissolved in Et2O, followed by adding NIS, DTBMP (1 equiv),
5, and TMSOTf (0.6 equiv) in turn, anomeric mixture of glycosides
3 and 8 was produced after 72 h without racemization. Meanwhile,
the anomeric selectivity was raised to 9:1 (entry 2). Obviously,
DTBMP served as an addictive prolonged the reaction time but im-
proved the a-selectivity. We supposed that DTBMP can effectively
suppress Lewis acid promoters to make the coupling reaction occur
in a mild condition.
Considering the homogeneous acyl chains ligated at C60, C1, and
C2, precursor 2 was reasonably designed first. The structure of 2
would enable us to acylate the C60 amino and C1, C2 hydroxyl
groups with corresponding acyl chloride simultaneously. Thus a
properly protected glycosyl glycerol derivative 3 was envisaged
as a key intermediate, whose C60 was linked with azide as a latent
amino group and C1, C2 hydroxyl groups were protected with
acetonide. To get this key intermediate 3, we assumed that an
appropriate glycosyl donor 4 with the 60-azido group and non-
participating benzyl group at C20 could couple with (S)-1,2-isopro-
To further improve the stereoselectivity and efficiency of glyco-
sylation, we paid attention to the Schmidt method which usually
worked well for the synthesis of
a
-glycoside with good yield.18
pylideneglycerol 5 to construct the
a
-glycosidic bond.
The glycosylation of (S)-isopropylideneglycerol with trichloroace-
timidate donor 4b was subsequently investigated (Table 1, entries
3–5). Compared to the thioglycoside 4a, this acid-catalyzed reaction
proceeded quickly without the racemization of the (S)-isopropylid-
eneglycerol residue. We found this process, which employed
chemically active imidate donor 4b with low substrate concentra-
The initial effort was to build up the donor 4 as shown in
Scheme 2. Thioglycoside 6 with the 6-hydroxyl free was synthe-
sized in five steps from glucose penta-acetate according to the lit-
erature procedures.15 Treatment of 6 with p-toluene sulfonyl
chloride produced compound 7, which was immediately reacted
with sodium azide in DMF to yield the thioglycoside 4a as the first
donor. Removal of the p-tolylthio group using N-bromosuccini-
mide (NBS) in acetone-water, followed by treatment with trichlo-
roacetonitrile (CCl3CN) and 1,8-diazabicyclo[5.4.0]-undec-7-ene
(DBU) gave trichloroacetimidate 4b as the second donor.
tion, can achieve good
a-selectivity in glycosylation. A high stere-
oselectivity ( /b = 33:1) could be achieved when the concentration
a
of 4b was 20 mM (entry 5). The trace amount of b anomer can be
isolated easily by silica gel column chromatography in the next
step. When the concentration of 4b was enhanced to 40 mM and
With the donors 4a and 4b available, glycosylation of (S)-iso-
propylideneglycerol 5 with the donors was then carefully exam-
ined. The results are summarized in Table 1. The main challenge
80 mM, the stereoselectivity would decline to
a/b 8:1 (entry 4)
and /b 4:1 (entry 3), respectively. The concentration of the reac-
a
tion mixture influenced anomeric selectivity and the reason was
presumably explained that mild conditions were beneficial for
1,2-cis glycosylation.19 However, this high stereoselectivity with
low concentration effect for imidate donor 4b was just opposite
to the study of using chemically stable thioglycoside donors in
appropriate CH2Cl2/nitrile solvent mixture.20
in the glycosylation was to efficiently construct the
a-configura-
tion glycosidic bond. Firstly, the glycosylation of (S)-isopropylid-
eneglycerol with donor 4a was carried out in the presence of
N-iodosuccinimide (NIS)–trimethylsilyl trifluoromethanesulfonate
(TMSOTf) in absolute ether (entry 1). This reaction was completed
in 4 h and gave a mixture of
their C2-epimers. The ratio of
a
-glycoside 3 and b-glycoside 8 and
/b (5:1) was determined by the
With the key intermediate 3 in hand, we accessed the final com-
pounds through four steps (Scheme 3). Hydrolysis of the isopropyl-
idene protecting group with TsOH in methanol provided 9 in 93%
yield. Then reduction of the azido group by Staudinger reaction
yielded the amino derivative 2. Introduction of acyl chains on C1,
C2, and C60–NH2 of 2 by condensation with corresponding acyl
chloride in the presence of 4-N,N-dimethylaminopyridine (DMAP)
gave 10a–h in 66–92% yield. Then removal of the benzyl by
hydrogenolysis using H2/Pd(OH)2 generated the final compounds
1a–h. All structures of the synthetic compounds were identified
a
1H NMR spectrum. Under such coupling conditions, we observed
the isopropylidene migration which resulted in the glycoside prod-
ucts with undesired C2-racemization. Intramolecular migration of
the isopropylidene group before glycosylation readily occurred un-
der Lewis acids.16 Danishefsky reported that the glycosylation of
glycosyl fluoride with 5 in the presence of 2,6-di-tert-butylpyridine
(DTBP) can prevent the racemization.17 Here the hindered base 2,6-
di-tert-butyl-4-methylpyridine (DTBMP) was employed. When 4a
OTs
O
OH
O
N3
O
N3
O
a
c
b
BnO
BnO
BnO
BnO
BnO
BnO
BnO
BnO
STol
STol
STol
OBn
OBn
OBn
4a
OBn
CCl3
NH
O
7
6
4b
Scheme 2. Synthesis of the glycosyl donors. Reagents and conditions: (a) CH2Cl2, Et3N, DMPA, TsCl, 95%; (b) NaN3, DMF, 65 °C, 88%; (c) NBS, acetone/water (9:1); then DBU,
CNCCl3, CH2Cl2, 76% over two steps.

8
Y. Sun et al. / Carbohydrate Research 355 (2012) 6–12
Table 1
Glycosylation of 4a and 4b with 5a
N3
O
BnO
BnO
STol
N3
O
OBn
N3
HO
4a
N3
O
O
O
O
BnO
BnO
BnO
BnO
5
O
O
+
O
OBn
OBn
O
BnO
BnO
O
promoters
O
8
3
OBn
O
CCl3
NH
Promoter (equiv)
4b
Entry
Donor
Temp
Time (h)
Yield (%)
Ratio (
a
/b)b
C2-chiralityc
1
4a
4a
4b
4b
4b
NIS–TMSOTf (1.5–0.6)
NIS–TMSOTf (1.5–0.6)
TMSOTf (0.2)
TMSOTf (0.2)
TMSOTf (0.2)
rt
rt
0 °C
0 °C
0 °C
4
86
85
87
87
88
5:1d
9:1
4:1
8:1
33:1
R
2e
3f
4g
5h
72
0.5
0.5
0.5
NR
NR
NR
NR
a
b
c
All reactions were carried out in absolute ether with 1.2 equiv acceptor.
Determined by 600 MHz 1H NMR.
R = Racemation, NR = No racemization.
C-2 epimers were not shown.
1 equiv DTBMP was added.
Concentration of 4b was 80 mM.
d
e
f
g
h
Concentration of 4b was 40 mM.
Concentration of 4b was 20 mM.
N3
NH2
O
N3
O
O
BnO
BnO
a
BnO
BnO
b
OH
BnO
OH
BnO
O
OBn
OBn
OBn
9
O
O
OH
O
OH
O
3
2
a: R = palmitoyl
b: R = hexanoyl
c: R = octanoyl
d: R = lauroyl
NHR
O
NHR
O
c
d
BnO
BnO
HO
HO
OR
OR
e: R = myristoyl,
OBn
OH
f: R = stearoyl
OR
OR
O
O
g: R = isovaleryl
10
1a-h
h: R = hydrocinnamoyl
Scheme 3. Synthesis of the target compounds 1a–h. (a) TsOH, MeOH, 93%; (b) PPh3, THF–H2O, 96%; (c) acyl chloride, Py, DMAP; (d) Pd(OH)2/C, H2, THF/i-PrOH (9:1).
by 1H, 13C NMR, and MS. And the data of 1a were in accord with the
rotation was determined with a Perkin–Elmer Model 241 MC
polarimeter. 1H NMR and 13C NMR spectra were taken on a JEOL
JNM-ECP 600 spectrometer with tetramethylsilane (Me4Si) as the
internal standard, and chemical shifts were recorded as d values.
Mass spectra were recorded on a Global Q-TOF mass spectrometer
and IonSpec 4.7 T FTMS (MALDI/DHB).
data of the natural product reported before.6,12
3. Conclusion
The natural aminoglycoglycerolipid 1a and its analogues were
synthesized in an efficient method with high stereoselectivity.
The glycosylation reaction employed trichloroacetimidate donor
4.2. p-Tolyl-2,3,4-tri-O-benzyl-6-O-p-tosyl-1-thio-b-D-glucopy
4b at low substrate concentration (20 mM) can achieve high
a-
ranoside (7)
configuration selectivity ( /b = 33:1). The present synthesis also
a
provided various acyl derivatives of glycolipid 1a required for the
studies on the structure–activity relationship and on the mode of
action. A detailed SAR study of these analogues will be reported
later.
To a mixture of 6 (1.1 g, 2.0 mmol) and TsCl (1.5 g, 8.0 mmol) in
CH2Cl2 (20 mL) were added Et3N (0.55 mL, 4.0 mmol) and catalytic
amount of DMAP. The reaction mixture was stirred at room tem-
perature for 5 h, and then washed with aq HCl (1 M) and brine.
The organic phase was collected, dried with Na2SO4, and concen-
trated. The residue was purified by column chromatography (8:1
petroleum ether–EtOAc) to provide 7 (1.32 g, 95%) as a white solid;
1H NMR (600 MHz, CDCl3) d 7.06–7.80 (m, 23H, Ar) , 4.88 (d, 1H, J
11.0 Hz, PhCH), 4.87 (d, 1H, J 10.4 Hz, PhCH), 4.81 (d, 1H, J 11.0 Hz,
PhCH), 4.80 (d, 1H, J 11.0 Hz, PhCH), 4.68 (d, 1H, J 10.4 Hz, PhCH),
4.52 (d, 1H, J 10.7 Hz, PhCH), 4.50 (d, 1H, J 9.8 Hz, H-1), 4.25 (dd,
1H, J 10.4, 1.6 Hz, H-60), 4.17(dd, 1H, J 10.4, 4.4 Hz, H-6), 3.64 (t,
1H, J 8.8 Hz, H-3), 3.48 (t, 1H, J 8.3 Hz, H-4), 3.45 (ddd, 1H, J 9.9,
4. Experimental
4.1. General methods
Solvents were purified in a conventional manner. Thin layer
chromatography (TLC) was performed on precoated HSGF254
plates (Yantai, China). Flash column chromatography was
performed on silica gel (200–300 mesh, Qingdao, China). Optical

Y. Sun et al. / Carbohydrate Research 355 (2012) 6–12
9
4.4, 1.7 Hz, H-5), 3.39 (dd, 1H, J 9.7, 8.9 Hz, H-2), 2.40 (s, 3H,
ArCH3), 2.33 (s, 3H, ArCH3); 13C NMR (150 MHz, CDCl3) d 21.3,
21.8, 68.6, 75.3, 75.6, 76.0, 76.6, 77.0, 80.7, 86.7, 87.6, 127.9–
130.1, 133.0, 133.1, 133.2, 137.7, 138.1, 138.2, 138.4, 145.1; HR-
ESI-MS m/z Calcd for C41H43O7S2 711.2445 [M+H]+, C41H42O7NaS2
733.2264 [M+Na]+. Found 711.2452 [M+H]+, 733.2271 [M+Na]+.
ished pressure and the residue was dissolved in CH2Cl2. The solu-
tion was washed with 10% Na2S2O3, followed by satd aq NaHCO3.
The organic layer was dried over Na2SO4, then the solvent was re-
moved in vacuo, and the residue was purified by column chroma-
tography (8:1 petroleum ether–EtOAc) to afford a mixture of 3 and
8 (0.36 g, 85%,
a
/b = 9:1) as a syrup; 1H NMR (CDCl3, 600 MHz): d
H-1 for 3: 4.86 (d, J 3.8 Hz), d H-1 for 8: 4.46 (d, J 7.7 Hz).
4.3. p-Tolyl-6-azido-2,3,4-tri-O-benzyl-6-deoxy-1-thio-b-D-
glucopyranoside (4a)
4.5.2. For donor 4b (entry 5)
A solution of trichloroacetimidate 4b (60.0 mg, 0.097 mmol)
A mixture of 7 (1.0 g, 1.4 mmol) and sodium azide (0.46 g,
7.0 mmol) in dry DMF (15 mL) was stirred at 60 °C overnight. After
the reaction was completed, the solvent was removed in vacuo and
the residue was dissolved in EtOAc. The excess of sodium azide and
sodium tosylate was removed by filtration. The filtrate was evapo-
rated and purified by silica gel column (12:1 petroleum ether–
and (S)-1,2-isopropylideneglycerol 5 (15
Et2O (4.9 mL) was added to freshly dried powdered 4 Å MS and
cooled to 0 °C. TMSOTf (3.9 L, 0.019 mmol) was added to the solu-
tion, and the mixture was stirred at 0 °C for 0.5 h. The mixture was
diluted with EtOAc (50 mL) and filtered through Celite. The organic
layer was washed with satd aq NaHCO3 and brine, dried (MgSO4),
and concentrated. The residue was purified by column chromatog-
raphy on silica gel (8:1 petroleum ether–EtOAc) to furnish 3 and 8
lL, 0.116 mmol) in dry
l
EtOAc) to afford 4a (0.73 g, 88%) as a white solid; ½a D22
¼ þ21:8 (c
ꢀ
0.70, CHCl3); 1H NMR (CDCl3, 600 MHz): d 7.11–7.49 (m, 19H,
Ar), 4.58–4.93 (m, 6H, 3ꢁ PhCH2), 4.73 (d, 1H, J 9.9 Hz, H-1), 3.68
(t, 1H, J 8.8 Hz, H-3), 3.52 (dd, 1H, J 13.2, 2.2 Hz, H-6), 3.45–3.50
(m, 2H, H-2, H-4), 3.40–3.43 (m, 1H, H-5), 3.33 (dd, 1H, J 13.2,
5.5 Hz, H-60), 2.34 (s, 3H, ArCH3); 13C NMR (150 MHz, CDCl3) d
21.4, 51.6, 75.4, 75.6, 76.0, 78.3 (2C), 81.0, 86.8, 88.2, 128.0–
128.8, 130.0, 133.6, 137.9, 138.2, 138.4; LR-ESI-MS m/z Calcd for
[M+Na]+ 604.2. Found 604.3.
(49.7 mg, 88%,
a/b = 33:1) as a colorless oil; for a anomer (3):
½
a 2D2
ꢀ
¼ þ58:0 (c 0.50, CHCl3), lit.13 +58.5 (c 11.96, CHCl3); 1H NMR
(CDCl3, 600 MHz): d 7.21–7.35 (m, 15H, –Ar); 4.55–5.0 (m, 6H,
3ꢁ PhCH2), 4.86 (d, 1H, J 3.8 Hz, H-1), 4.33–4.35 (m, 1H, Hsn-2),
0
4.07(dd, 1H, J 8.2, 6.0 Hz, Hsn-3 ), 3.96 (t, 1H, J 9.4 Hz, H-3), 3.80–
3.84 (m, 1H, H-5), 3.74 (dd, 1H, J 8.2, 6.0 Hz, Hsn-3), 3.63 (dd, 1H,
0
J 11.0, 6.0 Hz, Hsn-1 ), 3.53–3.57 (m, 2H, H-2, Hsn-1), 3.42–3.47 (m,
2H, H-4, H-60), 3.33 (dd, 1H, J 13.2, 5.5 Hz, H-6), 1.37 (s, 3H,
–CH3), 1.43 (s, 3H, –CH3); HR-ESI-MS m/z Calcd for C33H39O7N3Na
612.2680 [M+Na]+. Found 612.2671.
4.4. 6-Azido-2,3,4-tri-O-benzyl-6-deoxy-a-D-glucosyl trichloro
acetimidate (4b)
To a stirred solution of 4a (1.2 g, 2.0 mmol) in acetone (36 mL)
and H2O (4 mL) at ambient temperature was added NBS (1.1 g,
6.0 mmol). After 5 min, the reaction was quenched with saturated
NaHCO3 and the mixture concentrated. The crude intermediate
was diluted with CHCl3 (150 mL) and washed with satd aq NaHCO3
(100 mL), brine (2 ꢁ 100 mL), dried over Na2SO4, and concentrated.
The residue was purified by silica gel column chromatography to
afford white solid (0.87 g). To a stirred solution of the white solid
4.6. 3-O-(60-Azido-20,30,40-tri-O-benzyl-60-deoxy-
sn-glycerol (9)
a-D-glucosyl)-
TsOH (0.32 g, 1.70 mmol) was added to a stirred solution of 3
(0.50 g, 0.85 mmol) in MeOH (20 mL). After stirring for 2 h at ambi-
ent temperature, the solution was concentrated and dissolved in
CH2Cl2. The organic layer was washed with satd aq NaHCO3 and
water, dried over Na2SO4, filtered, and concentrated. Purification
was achieved by silica gel column chromatography (2:1 petroleum
ether–EtOAc) to give 9 (0.27 g, 93%) as a colorless waxy substance;
were added CCl3CN (1.1 mL, 11.0 mmol) and DBU (137 lL,
0.92 mmol) in CH2Cl2 (20 mL) at 0 °C. After 4 h, the solution was
concentrated and the residue was purified by silica gel column
chromatography (10:1 petroleum ether–EtOAc) to afford 4b
(0.97 g, 76%) as a syrup; 1H NMR (CDCl3, 600 MHz): d 8.62 (s, 1H,
NH), 7.24–7.34 (m, 15H, Ph), 6.50 (d, 1H, J 4.4 Hz, H-1), 4.60–4.99
(m, 6H, 3ꢁ PhCH2), 4.05 (t, 1H, J 9.8 Hz, H-4), 4.01 (ddd, 1H, J 9.9,
4.4, 2.2 Hz, H-5), 3.74 (dd, 1H, J 9.9, 3.3 Hz, H-2), 3.60 (t, 1H, J
8.8 Hz, H-3), 3.53 (dd, 1H, J 13.2, 2.2 Hz, H-6), 3.37 (dd, 1H, J
13.2, 4.4 Hz, H-60); 13C NMR (150 MHz, CDCl3) d 51.1, 72.8, 73.2,
75.7, 75.9, 79.6, 81.3, 91.4, 94.0, 94.1, 128.2–128.8, 137.9, 138.0,
½
a 2D2
ꢀ
¼ þ44:2 (c 0.75, CHCl3), lit.13 +35.7 (c 0.56, CHCl3); 1H NMR
(CDCl3, 600 MHz): d 7.25–7.36 (m, 15H, –C6H5), 4.57–4.97 (m,
6H, 3ꢁ PhCH2), 4.73 (d, 1H, J 3.8 Hz, H-1), 3.95 (t, 1H, J 9.4 Hz, H-
3), 3.84–3.89 (m, 2H, Hsn-1, Hsn-2), 3.80–3.83 (m, 1H, H-5), 3.74
0
(dd, 1H, J 11.6, 4.4 Hz, Hsn-3), 3.64 (dd, 1H, J 11.5, 4.4 Hz, Hsn-3 ),
0
3.56 (dd, 1H, J 9.9, 3.8 Hz, H-2), 3.40–3.47 (m, 3H, H-4, H-6, Hsn-1 ),
3.33 (dd, 1H, J 12.7, 5.5 Hz, H-60); HR-ESI-MS m/z Calcd for
C
30H36O7N3 550.2548 [M+H]+, C30H35O7N3Na 572.2367 [M+Na]+.
Found 550.2537 [M+H]+, 572.2357 [M+Na]+.
138.6, 161.4; HR-ESI-MS m/z Calcd for
C29H29O5N4Cl3Na
641.1096 [M+Na]+. Found 641.1078.
4.7. 3-O-(60-Amino-20,30,40-tri-O-benzyl-60-deoxy-
sn-glycerol (2)
a-D-glucosyl)-
4.5. 3-O-(60-Azido-20,30,40-tri-O-benzyl-60-deoxy-
1,2-isopropylidene -sn-glycerol (3)
a-D-glucosyl)-
To a solution of 9 (0.60 g, 1.08 mmol) in THF (20 mL) and water
(0.21 mL) was added PPh3 (0.57 g, 2.16 mmol). The reaction mix-
ture was heated at 50 °C for 5 h and then cooled to room temper-
ature. The mixture was evaporated under diminished pressure, and
the residue was purified by gel silica column chromatography
(10:1 CH2Cl2–MeOH) to give 2 (0.55 g, 96%) as a colorless waxy
substance; 1H NMR (CDCl3, 600 MHz): d 7.24–7.34 (m, 15H,
–C6H5), 4.55–4.96 (m, 6H, 3ꢁ PhCH2), 4.73 (d, 1H, J 3.7 Hz, H-1),
3.93 (t, 1H, J 9.2 Hz, H-3), 3.86–3.89 (m, 1H, Hsn-2), 3.69–3.73 (m,
4.5.1. For donor 4a (entry 2)
A mixture of the glycosyl donor 4a (0.41 g, 0.70 mmol) and acti-
vated 4 Å molecular sieves was stirred in dry Et2O (40 mL) at room
temperature under an N2 atmosphere, then NIS (0.23 g, 1.05 mmol)
was added. After stirring for 10 min, DTBMP (143 mmg, 3.8 mmol)
was added, followed by addition of acceptor (S)-1,2-isopropylid-
0
eneglycerol (100
l
L, 0.84 mmol) and TMSOTf (150
l
L, 0.42 mmol).
2H, Hsn-3, Hsn-3 ), 3.62–3.67 (m, 2H, H-5, Hsn-1), 3.50 (dd, 1H, J 9.6,
0
The reaction mixture was stirred at room temperature under an N2
atmosphere for 72 h, when TLC showed that the reaction was com-
pleted. The mixture was quenched by addition of Et3N, filtered
through a pad of Celite. The filtrate was concentrated by dimin-
3.7 Hz, H-2), 3.45 (dd, 1H, J 10.1, 6.4 Hz, Hsn-1 ), 3.20–3.26 (m, 3H,
H-4, –NH2), 2.97 (dd, 1H, J 12.8, 2.3 Hz, H-6), 2.61 (dd, 1H, J 12.8,
8.3 Hz, H-60); 13C NMR (150 MHz, CDCl3) d 42.4, 63.0, 70.1, 70.2,
71.7, 73.6, 75.2, 75.9, 79.4, 80.2, 82.3, 97.5, 128.2–128.6, 138.0,

10
Y. Sun et al. / Carbohydrate Research 355 (2012) 6–12
138.8; HR-ESI-MS m/z Calcd for C30H38O7N 524.2643 [M+H]+.
173.6; HR-ESI-MS m/z Calcd for C54H80O10N 902.5777 [M+H]+.
Found 524.2634.
Found 902.5788.
4.8. General procedure for 10a–h formation
4.8.4. 1,2-Dilauroyl-3-O-(N -lauroyl-60-amino-20,30,40-tri-O-
benzyl-60-deoxy-
a-D-glucosyl)-sn-glycerol (10d, 92%)
To a solution of compound 2 (80 mg, 0.15 mmol) in dry pyridine
(4 mL), a catalytic amount of DMAP and acyl chloride (6 equiv)
were added. The reaction mixture was stirred at room temperature
for 4 h, and then was diluted with EtOAc (15 mL) and washed with
satd aq NH4Cl (15 mL). The organic phase was dried over MgSO4,
filtrated, and concentrated. Purification by flash chromatography
(petroleum ether–EtOAc) yielded the corresponding compounds
10a–h (66–92%) as a colorless syrup or white solid.
1H NMR (CDCl3, 600 MHz): d 7.26–7.35 (m, 15H, –C6H5), 5.67
(dd, 1H, J 4.4, 7.7 Hz, –NH–CO–), 5.21–5.23 (m, 1H, Hsn-2), 4.61–
4.97 (m, 6H, 3ꢁ PhCH2), 4.69 (d, 1H, J 3.3 Hz, H-1), 4.39 (dd, 1H, J
0
3.8, 12.0 Hz, Hsn-3), 4.18 (dd, 1H, J 6.6, 12.1 Hz, Hsn-3 ), 3.95 (t, 1H,
J 9.3 Hz, H-3), 3.75–3.78 (m, 1H, H-5), 3.65–3.70 (m, 2H, H-6,
0
H
sn-1), 3.57 (dd, 1H, J 5.5, 10.4 Hz, Hsn-1 ), 3.47 (dd, 1H, J 3.8,
9.9 Hz, H-2), 3.32–3.35 (m, 1H, H-60), 3.27 (t, 1H, J 9.4 Hz, H-4),
2.28–2.34 (m, 4H, 2ꢁ –CH2–COO–), 2.11 (t, 2H, J 7.1 Hz, –NHCO–
CH2–), 1.57–1.64 (m, 6H, 3ꢁ –CO–CH2–CH2–), 1.21–1.33 (m, 48H,
3ꢁ –CH2–CH2(CH2)8–CH3), 0.87 (t, 9H, J 7.1 Hz, 3ꢁ –CH3); 13C
NMR (150 MHz, CDCl3) d 14.3, 22.9, 25.1, 26.0, 29.4–29.8, 32.1,
34.3, 34.5, 37.0, 39.7, 62.7, 66.8, 69.8, 70.1, 73.4, 75.5, 76.0, 78.8,
80.3, 81.7, 97.8, 128.0–128.7, 138.1, 138.4, 138.8, 173.3, 173.4,
173.6; HR-ESI-MS m/z Calcd for C66H104O10N 1070.7655 [M+H]+.
Found 1070.7647.
4.8.1. 1,2-Dipalmitoyl-3-O-(N -palmitoyl-60-amino-20,30,40-tri-O-
benzyl-60-deoxy-
a-D-glucosyl)-sn-glycerol (10a, 76%)
1H NMR (CDCl3, 600 MHz): d 7.26–7.35 (m, 15H, –C6H5), 5.66
(dd, 1H, J 7.3, 4.1 Hz, –NH–CO–), 5.21–5.23 (m, 1H, Hsn-2), 4.61–
4.97 (m, 6H, 3ꢁ PhCH2), 4.69 (d, 1H, J 3.3 Hz, H-1), 4.39 (dd, 1H, J
0
3.8, 12.1 Hz, Hsn-3), 4.18 (dd, 1H, J 6.1, 12.1 Hz, Hsn-3 ), 3.95 (t, 1H,
J 8.8 Hz, H-3), 3.75–3.78 (m, 1H, H-5), 3.65–3.70 (m, 2H, H-6,
4.8.5. 1,2-Dimyristoyl-3-O-(N -myristoyl-60-amino-20,30,40-tri-O-
0
H
sn-1), 3.57 (dd, 1H, J 5.5, 11.0 Hz, Hsn-1 ), 3.47 (dd, 1H, J 3.8,
benzyl-60-deoxy-
a-D-glucosyl)-sn-glycerol (10e, 83%)
9.9 Hz, H-2), 3.32–3.35 (m, 1H, H-60), 3.27 (t, 1H, J 9.4 Hz, H-4),
2.28–2.34 (m, 4H, 2ꢁ –CH2–COO–), 2.11 (t, 2H, J 7.1 Hz, –NHCO–
CH2–), 1.57–1.64 (m, 6H, 3ꢁ –CO–CH2–CH2–), 1.21–1.33 (m, 72H,
3ꢁ –CH2–CH2(CH2)12–CH3), 0.87 (t, 9H, J 6.6 Hz, 3ꢁ –CH3); 13C
NMR (150 MHz, CDCl3) d 14.3, 22.9, 25.0, 25.1, 26.0, 29.3–29.9,
32.1, 34.3, 34.5, 37.0, 39.8, 62.7, 66.8, 69.8, 70.1, 73.4, 75.5, 76.0,
78.9, 80.3, 81.7, 99.8, 128.1–128.7, 138.1, 138.4, 138.7, 173.4,
173.6, 179.0; HR-ESI-MS m/z Calcd for C78H128O10N 1238.9533
[M+H]+. Found 1238.9549.
1H NMR (CDCl3, 600 MHz): d 7.26–7.35 (m, 15H, –C6H5), 5.66
(dd, 1H, J 3.8, 7.1 Hz, –NH–CO–), 5.20–5.23 (m, 1H, Hsn-2), 4.61–
4.97 (m, 6H, 3ꢁ PhCH2), 4.69 (d, 1H, J 3.3 Hz, H-1), 4.39 (dd, 1H, J
0
3.8, 12.0 Hz, Hsn-3), 4.18 (dd, 1H, J 6.1, 11.5 Hz, Hsn-3 ), 3.95 (t, 1H,
J 8.8 Hz, H-3), 3.75–3.78 (m, 1H, H-5), 3.65–3.70 (m, 2H, H-6,
0
H
sn-1), 3.57 (dd, 1H, J 4.9, 10.4 Hz, Hsn-1 ), 3.47 (dd, 1H, J 3.8,
9.9 Hz, H-2), 3.32–3.35 (m, 1H, H-60), 3.27 (t, 1H, J 9.4 Hz, H-4),
2.28–2.35 (m, 4H, 2ꢁ –CH2–COO–), 2.11 (t, 2H, J 7.7 Hz, –NHCO–
CH2–), 1.57–1.64 (m, 6H, 3ꢁ –CO–CH2–CH2–), 1.21–1.33 (m, 60H,
3ꢁ –CH2–CH2(CH2)10–CH3), 0.87 (t, 9H, J 6.6 Hz, 3ꢁ –CH3); 13C
NMR (150 MHz, CDCl3) d 14.3, 22.9, 25.1, 26.0, 29.4–29.9, 32.1,
34.3, 34.5, 37.0, 39.8, 62.7, 66.8, 69.8, 70.1, 73.4, 75.5, 76.0, 78.8,
80.3, 81.7, 99.8, 128.1–128.6, 138.1, 138.3, 138.7, 173.4, 173.6,
178.4; HR-ESI-MS m/z Calcd for C72H116O10N 1154.8594 [M+H]+.
Found 1154.8600.
4.8.2. 1,2-Dihexanoyl-3-O-(N -hexanoyl-60-amino-20,30,40-tri-O-
benzyl-60-deoxy-
a-D-glucosyl)-sn-glycerol (10b, 81%)
1H NMR (CDCl3, 600 MHz): d 7.26–7.34 (m, 15H, –C6H5), 5.65
(dd, 1H, J 4.4, 7.7 Hz, –NH–CO–), 5.20–5.23 (m, 1H, Hsn-2), 4.62–
4.97 (m, 6H, 3ꢁ PhCH2), 4.69 (d, 1H, J 3.3 Hz, H-1), 4.39 (dd, 1H, J
0
3.9, 12.1 Hz, Hsn-3), 4.18 (dd, 1H, J 6.1, 11.6 Hz, Hsn-3 ), 3.95 (t, 1H,
J 9.4 Hz, H-3), 3.71–3.76 (m, 1H, H-5), 3.65–3.70 (m, 2H, H-6,
4.8.6. 1,2-Distearoyl-3-O-(N -stearoyl-60-amino-20,30,40-tri-O-
0
H
sn-1), 3.57 (dd, 1H, J 5.5, 11.0 Hz, Hsn-1 ), 3.47 (dd, 1H, J 3.3,
benzyl-60-deoxy-
a-D-glucosyl)-sn-glycerol (10f, 66%)
9.3 Hz, H-2), 3.33–3.36 (m, 1H, H-60), 3.27 (t, 1H, J 9.4 Hz, H-4),
2.28–2.31 (m, 4H, 2ꢁ –CH2–COO–), 2.11 (t, 2H, J 6.6 Hz, –NHCO–
CH2–), 1.57–1.62 (m, 6H, 3ꢁ –CO–CH2–CH2–), 1.26–1.31 (m, 12H,
3ꢁ –CH2–CH2(CH2)2–CH3), 0.88 (m, 9H, 3ꢁ –CH3); 13C NMR
(150 MHz, CDCl3) d 14.1, 14.2, 22.5, 22.6, 24.8, 25.7, 31.4–31.7,
34.3, 34.4, 37.0, 39.7, 62.6, 66.8, 69.8, 70.1, 73.4, 75.5, 76.0, 78.8,
80.3, 81.7, 97.8, 128.0–128.7, 138.1, 138.4, 138.8, 173.3, 173.4,
173.6; HR-ESI-MS m/z Calcd for C48H68O10N 818.4838 [M+H]+.
Found 818.4863.
1H NMR (CDCl3, 600 MHz): d 7.26–7.35 (m, 15H, –C6H5), 5.66
(dd, 1H, J 3.8, 7.1 Hz, –NH–CO–), 5.21–5.23 (m, 1H, Hsn-2), 4.61–
4.97 (m, 6H, 3ꢁ PhCH2), 4.69 (d, 1H, J 3.3 Hz, H-1), 4.39 (dd, 1H, J
0
3.3, 11.5 Hz, Hsn-3), 4.18 (dd, 1H, J 6.1, 12.1 Hz, Hsn-3 ), 3.95 (t, 1H,
J 9.4 Hz, H-3), 3.75–3.78 (m, 1H, H-5), 3.65–3.70 (m, 2H, H-6,
0
H
sn-1), 3.57 (dd, 1H, J 5.5, 11.0 Hz, Hsn-1 ), 3.47 (dd, 1H, J 3.3,
9.9 Hz, H-2), 3.32–3.35 (m, 1H, H-60), 3.27 (t, 1H, J 9.3 Hz, H-4),
2.28–2.34 (m, 4H, 2ꢁ –CH2–COO–), 2.11 (t, 2H, J 7.1 Hz, –NHCO–
CH2–), 1.57–1.64 (m, 6H, 3ꢁ –CO–CH2–CH2–), 1.21–1.33 (m, 84H,
3ꢁ –CH2–CH2(CH2)14–CH3), 0.87 (t, 9H, J 6.6 Hz, 3ꢁ –CH3); 13C
NMR (150 MHz, CDCl3) d 14.4, 22.9, 25.1, 26.0, 29.4–29.9, 32.2,
34.1, 34.3, 34.5, 37.0, 39.7, 62.7, 66.8, 69.8, 70.1, 73.4, 75.5, 76.0,
78.8, 80.3, 81.7, 97.8, 128.1–128.7, 138.1, 138.4, 138.8, 173.4,
173.6, 178.3; LR-APCI-MS: m/z Calcd for C84H140NO10 1323.0
[M+H]+. Found 1323.0.
4.8.3. 1,2-Dioctanoyl-3-O-(N -octanoyl-60-amino-20,30,40-tri-O-
benzyl-60-deoxy-
a-D-glucosyl)-sn-glycerol (10c, 86%)
1H NMR (CDCl3, 600 MHz): d 7.26–7.34 (m, 15H, –C6H5), 5.65
(dd, 1H, J 3.9, 7.1 Hz, –NH–CO–), 5.20–5.23 (m, 1H, Hsn-2), 4.62–
4.97 (m, 6H, 3ꢁ PhCH2), 4.69 (d, 1H, J 3.3 Hz, H-1), 4.39 (dd, 1H, J
0
3.8, 12.1 Hz, Hsn-3), 4.18 (dd, 1H, J 6.0, 12.1 Hz, Hsn-3 ), 3.95 (t, 1H,
4.8.7. 1,2-Diisovaleryl-3-O-(N -isovaleryl-60-amino-20,30,40-tri-O-
J 9.3 Hz, H-3), 3.75–3.78 (m, 1H, H-5), 3.65–3.70 (m, 2H, H-6,
benzyl-60-deoxy-
a-D-glucosyl)-sn-glycerol (10g, 78%)
0
H
sn-1), 3.57 (dd, 1H, J 5.5, 11.0 Hz, Hsn-1 ), 3.47 (dd, 1H, J 3.3,
9.4 Hz, H-2), 3.33–3.35 (m, 1H, H-60), 3.27 (t, 1H, J 9.9 Hz, H-4),
2.28–2.33 (m, 4H, 2ꢁ –CH2–COO–), 2.11 (t, 2H, J 7.1 Hz, –NHCO–
CH2–), 1.59–1.64 (m, 6H, 3ꢁ –CO–CH2–CH2–), 1.21–1.33 (m, 24H,
3ꢁ –CH2–CH2(CH2)4–CH3), 0.87 (t, 9H, J 7.1 Hz, 3ꢁ –CH3); 13C
NMR (150 MHz, CDCl3) d 14.3, 22.8, 25.1, 26.0, 29.1–29.4, 31.9,
34.3, 34.5, 37.0, 39.7, 62.6, 66.8, 69.8, 70.0, 73.4, 75.5, 76.0, 78.8,
80.3, 81.7, 97.8, 128.3–128.7, 138.1, 138.4, 138.7, 173.2, 173.4,
1H NMR (CDCl3, 600 MHz): d 7.26–7.34 (m, 15H, –C6H5), 5.63
(dd, 1H, J 3.3, 7.2 Hz, –NH–CO–), 5.21–5.24 (m, 1H, Hsn-2), 4.61–
4.96 (m, 6H, 3ꢁ PhCH2), 4.69 (d, 1H, J 3.3 Hz, H-1), 4.40 (dd, 1H, J
0
3.9, 12.1 Hz, Hsn-3), 4.18 (dd, 1H, J 6.1, 12.1 Hz, Hsn-3 ), 3.95 (t, 1H,
J 8.8 Hz, H-3), 3.72–3.76 (m, 1H, H-5), 3.65–3.71 (m, 2H, H-6,
0
H
sn-1), 3.57 (dd, 1H, J 5.5, 11.0 Hz, Hsn-1 ), 3.46 (dd, 1H, J 3.3,
9.9 Hz, H-2), 3.35–3.39 (m, 1H, H-60), 3.27 (t, 1H, J 9.4 Hz, H-4),

Y. Sun et al. / Carbohydrate Research 355 (2012) 6–12
11
1.95–2.19 (m, 9H, 3ꢁ –CO–CH2CH(CH3)2), 0.92–0.95 (m, 18H, 3ꢁ –
4.9.3. 1,2-Dioctanoyl-3-O-(N-octanoyl-60-amino-60-deoxy-
glucosyl)-sn-glycerol (1c, 90%)
a-D-
CO–CH2CH(CH3)2); 13C NMR (150 MHz, CDCl3) d 22.5, 22.6, 22.8,
25.8, 25.9, 26.3, 39.7, 43.3, 43.5, 46.3, 62.6, 66.8, 69.8, 70.0, 73.4,
75.4, 75.9, 78.9, 80.3, 81.7, 97.8, 128.2–128.7, 138.1, 138.4, 138.7,
172.6, 172.8; HR-ESI-MS m/z Calcd for C45H62O10N 776.4368
[M+H]+. Found 776.4375.
½
a 2D2
ꢀ
¼ þ15:9 (c 2.0, CHCl3); 1H NMR (CDCl3, 600 MHz): d 5.96
(dd, 1H, J 7.7, 4.4 Hz, –NH–CO–), 5.23–5.25 (m, 1H, Hsn-2), 4.81
(d, 1H, J 2.8 Hz, H-1), 4.39 (dd, 1H, J 3.8, 12.1 Hz, Hsn-1), 4.15 (dd,
0
1H, J 6.1, 12.1 Hz, Hsn-1 ), 3.86–3.91 (m, 1H, H-5), 3.77 (dd, 1H, J
5.0, 11.0 Hz, Hsn-3), 3.74 (t, 1H, J 9.4 Hz, H-3), 3.63 (dd, 1H, J 5.5,
4.8.8. 1,2-Dihydrocinnamoyl-3-O-(N -hydrocinnamoyl-60-
10.4 Hz, Hsn-3 ), 3.58 (dd, 1H, J 3.8, 9.4 Hz, H-2), 3.50 (dd, 1H, J
0
amino-20,30,40-tri-O-benzyl-60-deoxy-
a
-
D
-glucosyl)-sn-glycerol
3.8, 10.1 Hz, H-6), 3.13–3.18 (m, 2H, H-4, H-60), 2.21–2.32 (m,
6H, 3ꢁ –CH2–CO–), 1.61 (m, 6H, 3ꢁ –CO–CH2–CH2 –), 1.24–1.30
(m, 24H, 3ꢁ –CH2–CH2(CH2)4–CH3), 0.88 (t, 9H, J 7.7 Hz, 3ꢁ –
CH3); 13C NMR (150 MHz, CDCl3) d 14.2, 22.8, 25.0, 25.9, 29.1–
29.5, 31.8, 34.3, 34.5, 36.6, 40.1, 62.5, 66.8, 70.1, 70.6, 71.2, 72.4,
73.3, 99.6, 173.4, 173.7, 175.6; LR-ESI-MS m/z 632.5 [M+H]+,
654.5 [M+Na]+; HR-ESI-MS m/z Calcd for C33H62O10N 632.4374
[M+H]+, C33H61O10NNa 654.4193 [M+Na]+. Found 632.4387
[M+H]+, 654.4206 [M+Na]+.
(10h, 77%)
1H NMR (CDCl3, 600 MHz): d 7.10–7.34(m, 30H, –C6H5), 5.55
(dd, 1H, J 3.8, 7.1 Hz, –NH–CO–), 5.15–5.17 (m, 1H, Hsn-2), 4.56–
4.96 (m, 6H, 3ꢁ PhCH2), 4.61 (d, 1H, J 2.8 Hz, H-1), 4.34 (dd, 1H, J
0
3.8, 12.1 Hz, Hsn-3), 4.14 (dd, 1H, J 6.1, 12.1 Hz, Hsn-3 ), 3.90 (t, 1H,
J 9.4 Hz, H-3), 3.72–3.77 (m, 1H, H-5), 3.58–3.60 (m, 2H, H-6,
0
H
sn-1), 3.46 (dd, 1H, J 5.0, 11.0 Hz, Hsn-1 ), 3.38 (dd, 1H, J 3.8,
9.9 Hz, H-2), 3.24–3.27 (m, 1H, H-60), 3.14 (t, 1H, J 8.8 Hz, H-4),
2.40–2.95 (m, 12H, 3ꢁ –COCH2CH2–); 13C NMR (150 MHz, CDCl3)
d 31.0, 31.8, 35.7, 35.9, 38.5, 39.6, 62.8, 66.7, 69.7, 70.3, 73.3,
75.4, 75.9, 78.6, 80.2, 81.7, 97.8, 127.8–128.7, 138.1, 138.4, 138.8,
140.4, 140.5, 141.0, 172.2, 172.4, 172.5; HR-ESI-MS m/z Calcd for
4.9.4. 1,2-Dilauroyl-3-O-(N-lauroyl-60-amino-60-deoxy-
glucosyl)-sn-glycerol (1d, 86%)
a-D-
½
a 2D2
ꢀ
¼ þ10:5 (c 1.5, CHCl3); 1H NMR (CDCl3, 600 MHz): d 6.27
C
57H62O10N 920.4368 [M+H]+. Found 920.4385.
(dd, 1H, J 7.7, 4.9 Hz, –NH–CO–), 5.22–5.24 (m, 1H, Hsn-2), 4.81
(d, 1H, J 3.8 Hz, H-1), 4.39 (dd, 1H, J 3.3, 11.6 Hz, Hsn-1), 4.14 (dd,
0
4.9. General procedure for 1a–h formation
1H, J 6.1, 12.1 Hz, Hsn-1 ), 3.88–3.92 (m, 1H, H-5), 3.78 (dd, 1H, J
5.5, 11.0 Hz, Hsn-3), 3.74 (t, 1H, J 9.4 Hz, H-3), 3.63 (dd, 1H, J 5.5,
0
A solution of 10a–h (100 mg) in 20 mL THF/i-PrOH (9:1) was
treated with 10% palladium hydroxide (100 mg) and stirred at
ambient temperature under hydrogen atmosphere for 6 h. After fil-
tration the solvent was evaporated and the residue was purified by
column chromatography (CH2Cl2–MeOH) to afford 1a–h (84–90%)
as a colorless syrup or waxy solid.
11.0 Hz, Hsn-3 ), 3.58 (dd, 1H, J 3.8, 9.4 Hz, H-2), 3.48 (dd, 1H, J
3.8, 10.1 Hz, H-6), 3.13–3.16 (m, 2H, H-4, H-60), 2.22–2.32
(m, 6H, 3ꢁ –CH2–CO–), 1.61 (m, 6H, 3ꢁ –CO–CH2–CH2 –), 1.24–
1.31 (m, 48H, 3ꢁ –CH2–CH2(CH2)8–CH3), 0.88 (t, 9H, J 6.6 Hz, 3ꢁ
–CH3); 13C NMR (150 MHz, CDCl3) d 14.2, 22.7, 24.9, 25.8, 29.2–
30.0, 32.0, 34.2, 34.3, 36.5, 39.9, 62.4, 66.7, 70.2, 70.5, 71.1, 72.4,
73.2, 99.6, 173.3, 173.5, 175.5; LR-ESI-MS m/z 800.7 [M+H]+; HR-
4.9.1. 1,2-Dipalmitoyl-3-O-(N-palmitoyl-60-amino-60-deoxy-
glucosyl)-sn-glycerol (1a, 86%)
a
-
D
-
ESI-MS m/z Calcd for C45H86O10N
800.6252 [M+H]+. Found
800.6270.
½
a 2D2
ꢀ
¼ þ9:5 (c 0.75, CHCl3), lit.13 +12.4 (c 0.32, CHCl3); 1H NMR
(CDCl3, 600 MHz): d 5.95 (dd, 1H, J 7.7, 4.4 Hz, –NH–CO–), 5.22–
4.9.5. 1,2-Dimyristoyl-3-O-(N-myristoyl-60-amino-60-deoxy-
glucopyranosyl)-sn-glycerol (1e, 85%)
a-D-
5.24 (m, 1H, Hsn-2), 4.81 (d, 1H, J 3.8 Hz, H-1), 4.39 (dd, 1H, J 3.8,
0
11.5 Hz, Hsn-1), 4.13 (dd, 1H, J 6.1, 12.1 Hz, Hsn-1 ), 4.01–4.05 (m,
½
a 2D2
ꢀ
¼ þ6:2 (c 3.2, CHCl3); 1H NMR (CDCl3, 600 MHz): d 6.00
1H, H-5), 3.79 (dd, 1H, J 4.9, 11.0 Hz, Hsn-3), 3.75 (t, 1H, J 9.3 Hz,
(dd, 1H, J 8.2, 4.9 Hz, –NH–CO–), 5.23–5.25 (m, 1H, Hsn-2), 4.81
0
H-3), 3.63 (dd, 1H, J 6.1, 11.0 Hz, Hsn-3 ), 3.58 (dd, 1H, J 3.8,
(d, 1H, J 3.8 Hz, H-1), 4.39 (dd, 1H, J 3.8, 11.5 Hz, Hsn-1), 4.13 (dd,
0
9.5 Hz, H-2), 3.49 (dd, 1H, J 3.9, 11.1 Hz, H-6), 3.11 (t, 1H, J
9.4 Hz, H-4), 3.03–3.05 (m, 1H, H-60), 2.24–2.31 (m, 6H, 3ꢁ –CH2
–CO–), 1.60–1.65 (m, 6H, 3ꢁ –CO–CH2–CH2–), 1.26–1.30 (m, 72H,
3ꢁ –CH2–CH2(CH2)12–CH3), 0.87 (t, 9H, J 7.2 Hz, 3ꢁ –CH3); 13C
NMR (150 MHz, CDCl3) d 14.4, 22.9, 25.1, 25.9, 29.6–29.9, 32.2,
34.3, 34.5, 36.7, 40.0, 62.4, 67.1, 70.1, 70.3, 71.4, 72.6, 73.4, 99.7,
173.4, 173.7, 175.9; LR-APCI-MS m/z 968.9 [M+H]+; HR-MALDI-
1H, J 6.1, 12.1 Hz, Hsn-1 ), 3.97–4.01 (m, 1H, H-5), 3.78 (dd, 1H, J
4.4, 11.0 Hz, Hsn-3), 3.75 (t, 1H, J 9.3 Hz, H-3), 3.63 (dd, 1H, J 6.0,
11.0 Hz, Hsn-3 ), 3.58 (dd, 1H, J 3.8, 9.5 Hz, H-2), 3.49 (dd, 1H, J
0
4.4, 9.9 Hz, H-6), 3.12 (t, 1H, J 9.9 Hz, H-4), 3.04 (dd, 1H, J 7.4,
14.9 Hz, H-60), 2.24–2.34 (m, 6H, 3ꢁ –CH2 –CO–), 1.61–1.66 (m,
6H, 3ꢁ –CO–CH2–CH2–), 1.26–1.30 (m, 60H, 3ꢁ –CH2–
CH2(CH2)10–CH3), 0.88 (t, 9H,
J
7.2 Hz, 3ꢁ –CH3); 13C NMR
MS m/z Calcd for
C
57H109O10NNa 990.7949 [M+Na]+. Found
(150 MHz, CDCl3) d 14.3, 22.9, 25.0, 25.1, 25.9, 29.4–29.9, 32.2,
34.2, 34.3, 34.5, 36.7, 40.0, 62.4, 67.1, 70.1, 70.4, 71.3, 72.5, 73.3,
99.7, 173.5, 173.7, 176.0; LR-ESI-MS m/z 884.9 [M+H]+; HR-ESI-
MS m/z Calcd for C51H98O10N 884.7191 [M+H]+. Found 884.7210.
990.7922.
4.9.2. 1,2-Dihexanoyl-3-O-(N-hexanoyl-60-amino-60-deoxy-
glucosyl)-sn-glycerol (1b, 84%)
a-D-
½
a 2D2
ꢀ
¼ þ13:0 (c 0.90, CHCl3); 1H NMR (CDCl3, 600 MHz): d 6.06
4.9.6. 1,2-Distearoyl-3-O-(N-stearoyl-60-amino-60-deoxy-
glucosyl)-sn-glycerol (1f, 88%)
a-D-
(dd, 1H, J 6.1, 5.0 Hz, –NH–CO–), 5.16–5.18 (m, 1H, Hsn-2), 4.74 (d,
1H, J 2.2 Hz, H-1), 4.31 (dd, 1H, J 3.9, 12.1 Hz, Hsn-1), 4.08 (dd, 1H, J
½
a 2D2
ꢀ
¼ þ5:6 (c 1.2, CHCl3); 1H NMR (CDCl3, 600 MHz): d 5.89
0
6.1, 12.1 Hz, Hsn-1 ), 3.82–3.85 (m, 1H, H-5), 3.71 (dd, 1H, J 3.8,
(dd, 1H, J 8.7, 5.5 Hz, –NH–CO–), 5.22–5.25 (m, 1H, Hsn-2), 4.81
9.9 Hz, Hsn-3), 3.67 (t, 1H, J 9.9 Hz, H-3), 3.57 (dd, 1H, J 5.5,
(d, 1H, J 3.8 Hz, H-1), 4.39 (dd, 1H, J 3.8, 12.1 Hz, Hsn-1), 4.13 (dd,
0
0
10.4 Hz, Hsn-3 ), 3.51 (dd, 1H, J 3.8, 9.4 Hz, H-2), 3.41 (dd, 1H, J
1H, J 6.1, 12.1 Hz, Hsn-1 ), 3.97–4.01 (m, 1H, H-5), 3.78 (dd, 1H, J
5.0, 11.0 Hz, H-6), 3.06–3.09 (m, 2H, H-4, H-60), 2.13–2.26 (m,
6H, 3ꢁ –CH2–CO–), 1.54 (m, 6H, 3ꢁ –CO–CH2–CH2 –), 1.17–1.25
(m, 12H, 3ꢁ –CH2–CH2(CH2)2–CH3), 0.83 (t, 9H, J 7.1 Hz, 3ꢁ –
CH3); 13C NMR (150 MHz, CDCl3) d 14.1, 22.8, 25.0, 25.9, 29.1–
29.5, 31.8, 34.3, 34.5, 36.6, 40.1, 62.5, 66.8, 70.1, 70.6, 71.2, 72.4,
73.3, 99.6, 173.4, 173.7, 175.6; LR-ESI-MS m/z 548.2 [M+H]+; HR-
4.9, 11.0 Hz, Hsn-3), 3.75 (t, 1H, J 9.3 Hz, H-3), 3.63 (dd, 1H, J 6.5,
0
11.5 Hz, Hsn-3 ), 3.58 (dd, 1H, J 3.8, 9.9 Hz, H-2), 3.49 (dd, 1H, J
3.8, 10.4 Hz, H-6), 3.12 (t, 1H, J 9.9 Hz, H-4), 3.07 (m, 1H, H-60),
2.24–2.34 (m, 6H, 3ꢁ –CH2 –CO–), 1.61–1.66 (m, 6H, 3ꢁ –CO–
CH2–CH2 –), 1.26–1.30 (m, 84H, 3ꢁ –CH2–CH2(CH2)14–CH3), 0.87
(t, 9H, J 6.6 Hz, 3ꢁ –CH3); 13C NMR (150 MHz, CDCl3) d 14.3, 22.9,
25.0, 25.1, 25.9, 29.4–29.9, 32.2, 34.2, 34.3, 34.5, 36.7, 40.0, 62.4,
67.1, 70.1, 70.4, 71.3, 72.5, 73.3, 99.7, 173.5, 173.7, 176.0;
ESI-MS m/z Calcd for
548.3447.
C27H50O10N
548.3435 [M+H]+. Found

12
Y. Sun et al. / Carbohydrate Research 355 (2012) 6–12
LR-APCI-MS m/z 1052.9 [M+H]+; HR-MALDI-MS m/z Calcd for
63H121O10NNa 1074.8888 [M+Na]+. Found 1074.8889.
for Marine Scientific Research in the Public Interest (No.
201005024) and PCSIRT (No. IRT09444).
C
4.9.7. 1,2-Diisovaleryl-3-O-(N-isovaleryl-60-amino-60-deoxy-
a-
Supplementary data
D
-glucosyl)-sn-glycerol (1g, 88%)
½
a 2D2
ꢀ
¼ þ16:5 (c 1.5, CHCl3); 1H NMR (CDCl3, 600 MHz): d 6.25
Supplementary data associated with this article can be
found, in the online version, at http://dx.doi.org/10.1016/j.carres.
2012.04.005.
(dd, 1H, J 6.6, 5.0 Hz, –NH–CO–), 5.24–5.26 (m, 1H, Hsn-2), 4.81 (d,
1H, J 3.8 Hz, H-1), 4.40 (dd, 1H, J 3.3, 11.6 Hz, Hsn-1), 4.16 (dd, 1H,
0
J 6.1, 11.6 Hz, Hsn-1 ), 3.83–3.89 (m, 1H, H-5), 3.78 (dd, 1H, J 4.9,
10.4 Hz, Hsn-3), 3.74 (t, 1H, J 8.8 Hz, H-3), 3.63 (dd, 1H, J 5.5,
References
0
11.0 Hz, Hsn-3 ), 3.58 (dd, 1H, J 3.8, 9.9 Hz, H-2), 3.48 (dd, 1H, J 3.8,
10.1 Hz, H-6), 3.15–3.22 (m, 2H, H-4, H-60), 2.07–2.21 (m, 9H, 3ꢁ
–CO–CH2CH(CH3)2), 0.93–0.96 (m, 18H, –CO–CH2CH(CH3)2); 13C
NMR (150 MHz, CDCl3) d 22.6–22.7, 25.8, 25.9, 26.3, 40.1, 43.3,
43.5, 45.9, 62.5, 66.8, 70.0, 70.8, 71.2, 72.4, 73.3, 99.6, 172.7,
173.1, 174.9; LR-ESI-MS m/z 506.2 [M+H]+, 528.2 [M+Na]+; HR-
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amino-60-deoxy-
a-Diglucosyl)-sn-glycerol (1h, 87%)
½
a 2D2
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7.27 (m, 15H, C6H5), 6.04 (dd, 1H, J 7.2, 5.0 Hz, –NH–CO–), 5.17–
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0
12.1 Hz, Hsn-1), 4.09 (dd, 1H, J 5.5, 12.1 Hz, Hsn-1 ), 3.82–3.85 (m,
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Acknowledgements
This work was supported in parts by the National Natural Sci-
ence Foundation of China (No. 81072574), program for New Cen-
tury Excellent Talents in University (NCET-08-0506), Special Fund