Synthesis of 60-acylamido-60-deoxy-a-D-galactoglycerolipids

Article abstract of DOI:10.1016/j.carres.2013.02.008

Aminoglycoglycerolipid 1a isolated from an algal extract showed activity against the enzyme Myt1 kinase with an IC50 value of 0.12 lg/mL. Its analogues, 60-acylamido-60-deoxy-a-D-galactoglycerolipids (2a-g) were synthesized in an efficient way with high s

Full text of DOI:10.1016/j.carres.2013.02.008

Carbohydrate Research 376 (2013) 15–23
Contents lists available at SciVerse ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Synthesis of 60-acylamido-60-deoxy-
a-D-galactoglycerolipids
⇑
Chunxia Li , Yihua Sun, Jun Zhang, Zhimin Zhao, Guangli Yu, Huashi Guan
Key Laboratory of Marine Drugs, Ministry of Education, 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:
Aminoglycoglycerolipid 1a isolated from an algal extract showed activity against the enzyme Myt1
kinase with an IC50 value of 0.12 l a-D-galactoglycerolipids
g/mL. Its analogues, 60-acylamido-60-deoxy-
Received 15 December 2012
Received in revised form 28 January 2013
Accepted 17 February 2013
Available online 26 February 2013
(2a–g) were synthesized in an efficient way with high stereoselectivity. The key step was to employ a
4-OAc protecting group of the galactosyl donor 14 as a remote neighboring participation group to give
the glycoside with high
ity screening showed that compound 2g exhibited good inhibition against Myt1 kinase.
Ó 2013 Elsevier Ltd. All rights reserved.
a-anomeric selectivity (a:b = 32:1) in the glycosylation. The preliminary bioactiv-
Keywords:
Glycolipid
Myt1-kinase
Remote neighboring participation
a-Selectivity
1. Introduction
glycerolipid 1a were synthesized and tested in a binding assay to-
gether with a set of common kinase inhibitors against a full-length
Myt1 expressed in a human cell line.17 But these compounds
showed no significant effects in the binding assay.
The human Myt1 kinase is a Thr-14 and Tyr-15 specific regula-
tor of the cdc2/cyclin B kinase activity in the cell cycle. Inhibition of
Myt1 kinase is predicted to cause premature activation of cdc2.1,2
It has been reported that premature activation of cdc2 leads to mi-
totic catastrophe and cell death.1,3,4 So inhibitors of this enzyme
would thus be expected to kill rapidly proliferating cells and abro-
gate normal cell cycle checkpoints of cancer cells. Such inhibitors
would be attractive for the treatment of cancer in the future.
In 2005, aminoglycoglycerolipid 1a (Fig. 1), which was isolated
from an algal species showed high activity against the enzyme
For most of the bioactive natural glycoglycerolipids, the sugar
moiety was the galactosyl residue. Colombo et al. had found that
the axial configuration of the hydroxyl in the 4-position of the su-
gar of glycoglycerolipids was important for a remarkable anti-tu-
mor-promoting activity.14 These prompted us to synthesize a
series of 60-acylamido-60-deoxy-
a-D-galactoglycerolipids ana-
logues 2a–g (Fig. 1) by substituting the glucosyl with galactosyl
and altering the acyl chains, and ascertain the structural features
responsible for the activity. In this paper, we introduced an intelli-
gent method for the synthesis of these compounds and evaluated
their Myt1 kinase inhibition activity.
Myt1-kinase with an IC50 value of 0.12 l
g/mL.5 Due to the unique
structure and inhibition of Myt1-kinase, 1a had been synthesized
by our group and Schmidt et al.6–8
Glycoglycerolipids demonstrated various biological activities,
such as tumor growth and DNA polymerase inhibition,9,10 fatty
acid synthase inhibiton,11 glucose-lowing effect,12 and anti-inflam-
matory action.13 Colombo et al. had evaluated a series of glycoglyc-
erolipids for their anti-tumor promoting activity in vitro and
vivo.14–16 Reports showed that the minor structural changes of
the lipophilic acyl chain and the type of glycosyl in glycolipids af-
fected the antitumor activity of glycolipids. In order to acquire de-
tailed information of the structure–activity relationship (SAR) of
glycoglycerolipid 1a, we synthesized its analogues 1b–h (Fig. 1)
with different acyl chains.7 Recently, through changes in the con-
2. Results and discussion
The main challenge in the synthesis of 2a–g was to efficiently
construct the
a-configured glycosidic bond in the glycosylation.
The aminoglycoglycerolipids 1a–h have been synthesized with
high stereoselectivity by employing thioglycoside or trichloroace-
timidate as donor.7 Now we tried to verify whether the same syn-
thetic strategy could be applied to the synthesis of 2a–g.
Thiogalactoside 8a and trichloroacetimidate 8b were initially
investigated as the donors. Synthesis of 8a and 8b starting from
the known 1-thio-b-D
-galactopyranoside 318 was shown in
figuration of glycosidic bond (a, b), saccharide unit (glu, gal,
Man) and acyl chains (palmitoyl, capryloyl), 6 derivatives of glyco-
Scheme 1. Tritylation of the primary hydroxyl of the thiogalacto-
side 3 yielded the triol 4, which was perbenzylated to yield 5.
Acid-catalyzed removal of the trityl group provided the desired
alcohol 6 in an overall yield of 60% over the three steps.19 Treat-
⇑
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 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.carres.2013.02.008

16
C. Li et al. / Carbohydrate Research 376 (2013) 15–23
a
: R = palmitoyl
a: R = hexanoyl
b: R = octanoyl
b: R = hexanoyl
NHR
O
OH
NHR
O
c
: R = octanoyl
c
d
: R = lauroyl
d: R = lauroyl
HO
HO
: R = myristoyl
e
: R = myristoyl
HO
e: R = palmitoyl
f: R = stearoyl
OR
OR
f: R = stearoyl
OH
OH
g
: R = isovaleryl
O
O
g
: R = isovaleryl
OR
OR
h: R = hydrocinnamoyl
1
2
Figure 1. Structures of natural aminoglycoglycerolipid 1a and its analogues 1b–h and 2a–g.
ment of thiogalactoside 6 with p-toluenesulfonyl chloride pro-
duced tosylate 7. Then 7 reacted with sodium azide in DMF to af-
ford 8a in only 26% yield. We supposed that steric hindrance of 4-
OBn may influence the substitution by azide. Then 8a was con-
verted to trichloroacetimidate 8b following a standard procedure
to undertake the initial investigation.
Glycosylation of trichloroacetimidate 8b and (S)-isopropylid-
eneglycerol catalyzed by TMSOTf in absolute ether furnished a
2:5 inseparable mixture of 9a (H-1: d 4.93, J 3.8 Hz) and 9b (H-1:
OBn in the production of 8 (Scheme 1) was confirmed. Then 13 was
O-acetylated to afford the 6-azido-6-deoxy thiogalactoside donor
14.
Subsequently, the coupling reaction between 14 and the accep-
tor (S)-isopropylideneglycerol in various solvents was carefully
examined using NIS–TMSOTf as promoters at room temperature,
meanwhile, DTBMP was added to prevent (S)-isopropylideneglyc-
erol from racemization.24–27 The results were summarized in
Table 1.
d 4.39, J 7.7 Hz), whereas the glycosylation of thioglycoside 8a un-
der NIS–TMSOTf in the presence of DTBMP afforded a better ratio
(a/b = 9:5) without S-chiral center racemization (Scheme 2).
Racemization of the (S)-isopropylideneglycerol residue was not
observed in all the reactions, which were identified by 1H NMR.
First, the glycosylation of (S)-isopropylideneglycerol with donor
14 was carried out in dioxane/toluene (3:1) (entry 1). There was
no change in the reaction mixture during 48 h. When CH2Cl2
Although the ratio was improved, the donor thioglycoside 8a was
obtained in a low yield (Scheme 1), and the anomeric stereoselec-
tivity of the glycosylation was far from our expectation. So the
strategy used in the synthesis of glucosyl type aminoglycoglycer-
olipids 1a–h failed in the practical synthesis of 2a–g. But it was
encouraging that these results gave us a clue that a thioglycoside
may become a hopeful glycosyl donor for the stereoselective
glycosylation.
served as the solvent, products 15
92% yield with a modest /b ratio of 5:1 (entry 2). This stereoselec-
tivity was further raised to /b 25:1 when a mixed solvent CH2Cl2/
Et2O (1:3) was used (entry 3). The best result was achieved using
Et2O as solvent which gave almost exclusively the 15 isomer
(15 :15b = 32:1) (entry 4). These results supported the notion that
a and 15b were obtained in
a
a
a
a
Then a major effort was made to find a suitable galactosyl do-
the 4-OAc protecting group of the galactosyl donor can perform re-
nor, which should be designed to obtain a high
as a satisfactory yield for the preparation of -galactosyl glycerol.
Cox and Besra prepared 6-N-derivatized -galactosyl ceramides
a/b ratio as well
mote neighboring group participation to give galactosides with
a
high
effect.
Having completed the synthesis of 15
thesize the final compounds 2a–g (Scheme 4). The acetyl group
of 15 was converted to the benzyl group to produce 16. Hydroly-
a-anomeric selectivity, and that using Et2O can enhance this
a
using a glycosyl iodide donor, and found that small changes in
a, we continued to syn-
the sugar substitution pattern affected the donors’ activity
greatly.20 During a one-pot
a
-galactosylation method using Appel
agents in DMF, Nishida and Kobayashi showed that the donors
with the 6-O-acyl group can promote
-selectivity.21 Boons re-
a
sis of the isopropylidene group with TsOH and then reduction of
the azido group by the Staudinger reaction yielded the appropriate
amino derivative 17. Introduction of acyl chains on C1, C2 and C60–
NH2 of 17 by condensation with the corresponding acyl chloride in
the presence of 4-N,N-dimethylaminopyridine (DMAP) gave 18a–g
in 67–90% yield. Then removal of the benzyl groups by hydrogen-
olysis with H2/Pd(OH)2 generated the final compounds 2a–g. The
structures of the synthetic samples were identified by 1H, 13C
NMR spectroscopy, and HRESIMS.
Glycoglycerolipids 1a–h and 2a–g were assayed for inhibitory
activity against Myt1 kinase based on the chemiluminescence
method.28 The profiling data for various compounds against Myt1
protein kinase were shown in Table 2. Compared with the refer-
ence compound staurosporine and 1a, compound 2g showed good
inhibition activity. This result indicated that the galactosyl and iso-
valeryl groups exerted positive effect on the inhibition of Myt1 ki-
nase. From this table, we found that the activities of the galacto
a
ported that iodonium-ion promoted glycosylations in 1,4-diox-
ane/toluene with galactosyl donors having a neighboring group
participating functionality at C-4 gave exceptionally high
a-ano-
meric selectivity.22 Based on these facts, a new galactosyl donor
14 which was equipped with an acetyl group on the C-4 position
was rationally designed. The synthesis of 14 began from 3 in six
steps as shown in Scheme 3. Treatment of galactopyranoside 3
with benzaldehyde dimethyl acetal yielded the 4,6-O-benzylidene
derivative. The remaining free hydroxyl groups were benzylated to
afford compound 10 in 80% yield for the two steps. Then the ben-
zylidene group was removed to furnish 11 with the 4-OH and 6-OH
exposed. Compound 11 was tosylated with p-toluenesulfonyl chlo-
ride in pyridine to give 12 selectively in 70% yield.23 Treatment of
12 with sodium azide in DMF in the presence of 15-crown-5 gave
13 in 76% yield. Here our supposition of the steric hindrance of 4-
OH
OBn
OH
OBn
OH
OH
OTr
OTr
O
O
O
O
c
a
b
HO
STol
BnO
STol
HO
STol
N3
BnO
STol
N3
OH
OH
OBn
OBn
3
4
6
5
OBn
BnO
OBn
OBn
BnO
OTs
O
O
O
f
NH
CCl3
d
e
STol
BnO
STol
OBn
8b
OBn
OBn
O
7
8a
Scheme 1. Reagents and conditions: (a) TrCl, DMAP, Py, 80 °C; (b) BnBr, NaH, DMF, 66% for two steps; (c) AcOH–EtOH, 90 °C, 91%; (d) CH2Cl2, Et3N, DMAP, TsCl, 95%; (e) NaN3,
DMF, 65 °C, 26%; (f) NBS, acetone/water (9:1), then DBU, CNCCl3, CH2Cl2 72%.

C. Li et al. / Carbohydrate Research 376 (2013) 15–23
17
OBn
N3
O
STol
BnO
OBn
OBn
OBn
N3
N3
O
8a
O
O
O
O
BnO
BnO
+
O
or
OBn
OBn
8b
O
b for
O
9
9
β
α
OBn
N3
O
9α : 9β = 9 : 5 for 8a
9α : 9β = 2 : 5 for 8b
NH
BnO
OBn
O
CCl3
8b
Scheme 2. Reagents and conditions: (a) (S)-isopropylideneglycerol, DTBMP, NIS–TMSOTf, Et2O, 0 °C to rt, 5 h, 83%; (b) (S)-isopropyleneglycerol, TMSOTf, Et2O, 0 °C, 30 min,
86%.
Ph
O
OH
OH
BnO
O
OH
OH
O
O
O
c
a
b
HO
STol
STol
STol
STol
STol
BnO
OH
OBn
OBn
11
3
10
OH
BnO
N3
OH OTs
OAc
N3
O
O
O
e
d
BnO
STol
BnO
OBn
OBn
OBn
13
14
12
Scheme 3. Reagents and conditions: (a) PhCH(OCH3)2, DMF, TsOHꢀH2O, 50 °C, then BnBr, NaH, DMF, 80% for the two steps; (b) TsOHꢀH2O, MeOH, 50 °C, 88%; (c) TsCl, Py
ꢁ45 °C to rt, 70%; (d) NaN3, DMF, 15-crown-5, 65 °C, 76%; (e) Ac2O, CH2Cl2, Et3N, DMAP, 92%.
Table 1
Glycosylation of (S)-isopropylideneglycerol with 14 in various solventsa
OAc
OAc
OAc
N3
N3
N3
O
O
O
O
O
O
O
HO
O
STol
BnO
BnO
BnO
+
+
O
OBn
OBn
OBn
O
O
(S)-isopropylideneglycerol
15α
15β
14
Entry
Solvents
Time
Yieldb
Ratio (15a
:15b)c
1
2
3
4
Dioxane/toluene = 3:1
CH2Cl2
CH2Cl2/Et2O = 1:3
Et2O
—
—
—
3 h
5 h
16 h
92%
90%
90%
5:1
25:1
32:1
a
b
c
All reactions were carried out with 1.2 equiv (S)-isopropylideneglycerol in the presence of N-iodosuccinimide (1.5 equiv), TMSOTf (0.6 equiv) and DTBMP (1.0 equiv).
Isolated yield.
Determined by 600 MHz 1H NMR.
series were better than their gluco counterparts generally. We in-
ferred that the galactosyl residue performed positive effect on
the activity and a suitable acyl group would enforce this effect. Fur-
ther structural optimization and more detailed bioassay are
underway.
Et2O. The preliminary bioactivity screening showed that com-
pound 2g had good inhibition against Myt1 kinase. A detailed
structure antitumor activity relationship study of these analogues
will be reported later.
4. Experimental
3. Conclusions
4.1. General methods
In conclusion, a series of 60-acylamido-60-deoxy-
glycerolipids were synthesized in an efficient way with high stere-
oselectivity ( :b = 32:1) by employing 14 as a glycosyl donor in
a-D-galacto-
Solvents were purified in a conventional manner. The boiling
range of petroleum ether was 60–90 °C. Thin layer chromatogra-
a

18
C. Li et al. / Carbohydrate Research 376 (2013) 15–23
OAc
OBn
NH2
OBn
N3
N3
O
O
O
a
b
BnO
BnO
BnO
O
OH
OBn
O
OBn
OBn
O
O
O
O
OH
O
15
α
17
16
OH
a
NHR
: R = hexanoyl
OBn NHR
O
b: R = octanoyl
O
c
: R = lauroyl
d
c
HO
d: R = myristoyl
BnO
OR3
OR3
OH
e
: R = palmitoyl
OR
OBn
f: R = stearoyl
O
O
OR
18
2
g
: R = isovaleryl
Scheme 4. Reagents and conditions: (a) MeONa, MeOH, then BnBr, NaH, DMF, 82% for two steps; (b) TsOH, MeOH, then PPh3, THF–H2O, 88% for the two steps; (c) acyl
chloride, Py, DMAP, 67–90%; (d) Pd(OH)2/C, H2, THF/i-PrOH (9:1), 82–90%.
Table 2
4.50–4.88 (m, 6H, 3 ꢂ PhCH2), 4.71 (d, 1H, J 9.9 Hz, H-1), 3.90 (d,
Activity change (%) of Myt1 in the presence of various compounds
1H, J 2.7 Hz, H-4), 3.87 (t, 1H, J 9.4 Hz, H-2), 3.57 (dd, 1H, J 9.5,
6.0 Hz, H-6), 3.54 (dd, 1H, J 9.3, 2.7 Hz, H-3), 3.34 (app. t, J
6.4 Hz, 1H, H-5), 3.24 (dd, 1H, J 9.5, 6.7 Hz, H-60), 2.28 (s, 3H,
ArCH3); LR-ESI-MS m/z calcd for [M+Na]+ 821.3; found 821.3.
Compound
Activity change %
Compound
Activity change %
1b
1c
1d
1e
1a
1f
2
4
5
32
34
32
35
38
2a
2b
2c
2d
2e
2f
2g
9
15
24
23
42
40
60
65
4.2.2. p-Tolyl-2,3,4-tri-O-benzyl-1-thio-b-D-galactopyranoside
(6)29
Thioglycoside 5 (2.0 g, 2.51 mmol) was suspended in a mixture
of AcOH:EtOH (20 mL:10 mL) and refluxed at 90 °C for 16 h. Then
the reaction mixture was concentrated in vacuo (co-evaporation
with toluene) and the residue was dissolved in CH2Cl2 (200 mL),
washed sequentially with satd aq NaHCO3 (200 mL), brine
(200 mL), dried (Na2SO4) and concentrated in vacuo to afford alco-
hol 6 (1.26 g, 91%) as a white crystalline solid; 1H NMR (600 MHz,
CDCl3): d 7.01–7.52 (m, 19H, –Ar), 4.62–4.99 (m, 6H, 3 ꢂ PhCH2),
4.56 (d, 1H, J 9.7 Hz, H-1), 4.25 (dd, 1H, J 11.2, 7.0 Hz, H-6), 4.10
(dd, 1H, J 11.3, 5.6 Hz, H-60), 3.91 (t, 1H, J 9.4 Hz, H-2), 3.83 (d,
1H, J 2.1 Hz, H-4), 3.59 (dd, 1H, J 9.3, 2.7 Hz, H-3), 3.56 (app.t, J
6.3 Hz, 1H, H-5), 2.30 (s, 3H, ArCH3); LR-ESI-MS m/z calcd for
[M+Na]+ 579.2; found 579.2.
1g
1h
Staurosporine
phy (TLC) was performed on precoated HSGF254 plates (Yantai,
China). Flash column chromatography was performed on silica
gel (200–300 mesh, Qingdao, China). Optical rotation was deter-
mined with a Perkin–Elmer Model 241 MC polarimeter. IR spec-
troscopy was performed on
a
Nicolet Nexus 470 FT-IR
spectrometer using the KBr pellet technique. 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 Tesla FTMS
(MALDI/DHB).
4.2.3. p-Tolyl-2,3,4-tri-O-benzyl-6-O-p-tosyl-1-thio-b-D-
galactopyranoside (7)
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
DMAP (24 mg, 0.2 mmol). The reaction mixture was stirred at
room temperature for 5 h, and then washed sequentially with aq
HCl (1 M) and brine. The organic phase was collected, dried with
Na2SO4 and concentrated. The residue was purified by column
4.2. Synthetic chemistry
4.2.1. p-Tolyl-2,3,4-tri-O-benzyl-6-O-trityl-1-thio-b-D-
galactopyranoside (5)
To a mixture of tetrol 3 (5.3 g, 18.5 mmol) and trityl chloride
(7.7 g, 27.8 mmol) in pyridine (50 mL) was added catalytic DMAP
(0.23 g, 1.85 mmol). The reaction mixture was stirred at 80 °C for
5 h, and then concentrated in vacuo. The residue was dissolved
in CH2Cl2 (100 mL), washed sequentially with aq HCl (1 M,
2 ꢂ 100 mL), satd aq NaHCO3 (2 ꢂ 100 mL) and brine (100 mL).
The organic phase was collected, dried with Na2SO4 and concen-
trated in vacuo to afford the crude triol 4.
The crude residue 4 was dissolved in DMF (50 mL) and BnBr
(11 mL, 92.5 mmol) was added. The solution was cooled to 0 °C
and sodium hydride (4.5 g, 60%, 111 mmol) was added portionwise
to the stirred solution over 5 min. The reaction mixture was al-
lowed to warm to room temperature. After 6 h, MeOH (40 mL)
was added and the solution was stirred for 15 min. The reaction
mixture was then concentrated in vacuo (co-evaporation with tol-
uene, 3 ꢂ 50 mL) and the residue was dissolved in CH2Cl2 (100 mL).
The resulting solution was washed with brine (3 ꢂ 50 mL), dried
(Na2SO4) and concentrated in vacuo. The residue was purified by
flash column chromatography (1:12 EtOAc–petroleum ether) to af-
ford fully protected thioglycoside 5 (9.8 g, 66%) as a white crystal-
line solid; 1H NMR (600 MHz, CDCl3): d 6.95–7.46 (m, 34H, –Ar),
chromatography (8:1 petroleum ether–EtOAc) to provide
7
(1.32 g, 95%) as a white solid; 1H NMR (600 MHz, CDCl3): d 6.99–
7.74 (m, 23H, –Ar), 4.47–4.95 (m, 6H, 3 ꢂ PhCH2), 4.52 (d, 1H, J
9.6 Hz, H-1), 4.11 (dd, 1H, J 9.8, 6.2 Hz, H-6), 4.05 (dd, 1H, J 9.8,
6.7 Hz, H-60), 3.90 (d, 1H, J 2.2 Hz, H-4), 3.83 (t, 1H, J 9.4 Hz, H-2),
3.63 (app.t, J 6.4 Hz, 1H, H-5), 3.57 (dd, 1H, J 9.2, 2.6 Hz, H-3),
2.40 (s, 3H, ArCH3), 2.30 (s, 3H, ArCH3); LR-ESI-MS m/z calcd for
[M+Na]+ 733.2; found 733.3.
4.2.4. p-Tolyl-6-azido-2,3,4-tri-O-benzyl-6-deoxy-1-thio-b-D-
galactopyranoside (8a)30
A mixture of 7 (0.8 g, 1.1 mmol) and sodium azide (0.37 g,
5.5 mmol) in dry DMF (15 mL) was stirred at 65 °C overnight. Then
the solvent was removed in vacuo and the residue was dissolved in
EtOAc. The excess of sodium azide and sodium tosylate was re-
moved by filtration. The filtrate was evaporated and purified by sil-
ica gel column (12:1 petroleum ether–EtOAc) to afford 8a (0.17 g,
26%) as a white solid; 1H NMR (CDCl3, 600 MHz): d 7.04–7.48 (m,
19H, Ar), 4.60–5.03 (m, 6H, 3 ꢂ PhCH2), 4.56 (d, 1H, J 9.9 Hz, H-

C. Li et al. / Carbohydrate Research 376 (2013) 15–23
19
1), 3.89 (t, 1H, J 9.4 Hz, H-2), 3.79 (d, 1H, J 2.1 Hz, H-4), 3.58–3.62
(m, 2H, H-3, H-6), 3.43–3.45 (m, 1H, H-5), 3.15 (dd, 1H, J 12.1,
5.5 Hz, H-60), 2.32 (s, 3H, ArCH3); LR-ESI-MS m/z calcd for
[M+Na]+ 604.2; found 604.3.
(15 mL ꢂ 3). All the organic extracts were combined, dried over
Na2SO4 and concentrated. The residue was recrystallized in ethanol
to afford 10 (0.77 g, 80%) as a white solid; 1H NMR (600 MHz,
CDCl3): d 7.25–7.62 (m, 19H, –Ar), 5.48 (s, 1H, PhCH), 4.68–4.73
(m, 4H, 2 ꢂ PhCH2), 4.56 (d, 1H, J 9.3 Hz, H-1), 4.37 (dd, 1H, J
12.1, 1.6 Hz, H-6), 4.14 (d, 1H, J 2.8 Hz, H-4), 3.97 (dd, 1H, J 12.1,
1.7 Hz, H-60), 3.83 (t, 1H, J 9.3 Hz, H-2), 3.62 (dd, 1H, J 9.3, 3.3 Hz,
H-3), 3.40–3.42 (m, 1H, H-5), 2.30 (s, 3H, ArCH3); LR-ESI-MS m/z
calcd for [M+Na]+ 577.2; found 577.1.
4.2.5. 3-O-(60-azido-20,30,40-tri-O-benzyl-60-deoxy-
galactosyl)-1,2-isopropylidene-sn-glycerol (9
4.2.5.1. For donor 8a. A mixture of the glycosyl donor 8a
a-D-
a
)
(137 mg, 0.23 mmol) and freshly dried 4 Å molecular sieves was
stirred in dry Et2O (10 mL) at 0 °C under a N2 atmosphere, then
NIS (77 mg, 0.35 mmol) was added. After stirring for 10 min,
DTBMP (48 mg, 0.23 mmol) was added, followed by the addition
4.2.7. p-Tolyl-2,3-di-O-benzyl-1-thio-b-D-galactopyranoside (11)
A mixture of compound 10 (1.2 g, 2.1 mmol) and TsOHꢀH2O
(0.41 g, 2.1 mmol) in MeOH (30 mL) was stirred at room temper-
ature for 2 h. The reaction solution was diluted with EtOAc
(100 mL), and the mixture was consecutively washed by satd aq
NaHCO3 and brine. The organic layer was dried over NaSO4, fil-
tered, and concentrated in vacuo to get a residue, which was
recrystallized in EtOH to afford 11 (0.86 g, 88%) as a foamy solid;
1H NMR (600 MHz, CDCl3): d 7.02–7.39 (m, 14H, –Ar), 4.62–4.77
(m, 4 H, 2 ꢂ PhCH2), 4.51 (d, 1H, J 9.4 Hz, H-1), 3.97 (d, 1H, J
2.8 Hz, H-4), 3.88 (dd, 1H, J 11.5, 6.6 Hz, H-6), 3.72 (dd, 1H, J
11.5, 3.8 Hz, H-60), 3.64 (t, 1H, J 9.4 Hz, H-2), 3.50 (dd, 1H, J 8.8,
2.8 Hz, H-3), 3.39 (m, 1H, H-5), 2.24 (s, 3H, ArCH3), 2.08 (br s,
2H, 2 ꢂ –OH); LR-ESI-MS m/z calcd for [M+Na]+ 489.2; found
489.1.
of acceptor (S)-1,2-isopropylideneglycerol (34
lL, 0.28 mmol) and
TMSOTf (50 L, 0.14 mmol). The reaction mixture was stirred at
l
room temperature under a N2 atmosphere for 5 h, when TLC
showed that the reaction was complete. The mixture was
quenched by the addition of Et3N, filtered through a pad of Celite.
The filtrate was concentrated under reduced pressure and the res-
idue was dissolved in CH2Cl2. The solution was washed with 10%
Na2S2O3 solution, followed by satd aq NaHCO3. The organic layer
was dried over Na2SO4, then the solvent was removed in vacuo,
and the residue was purified by column chromatography (8:1
petroleum ether–EtOAc) to afford
(115 mg, 83%,
/b = 9:5) as a syrup; 1H NMR (CDCl3, 600 MHz): d
H-1 for 9
a mixture of 9a and 9b
a
a
: 4.93 (d, J 3.8 Hz), d H-1 for 9b: 4.40 (d, J 7.7 Hz). HR-
ESI-MS m/z calcd for C33H39O7N3 Na 612.2686 [M+Na]+, found
612.2680.
4.2.8. p-Tolyl-2,3-di-O-benzyl-6-O-p-tosyl-1-thio-b-D-
galactopyranoside (12)
4.2.5.2. For donor 8b.
To a stirred solution of 8a (120 mg,
To a solution of 11 (750 mg, 1.6 mmol) in dry pyridine (15 mL)
under N2 at ꢁ45 °C, was added TsCl (340 mg, 1.76 mmol). Then the
temperature was raised to ambient temperature. CH2Cl2 (30 mL)
was added after 6 h and the mixture was washed with water and
aq HCl (0.1 M). The organic phase was dried and concentrated
and the residue was purified by chromatography on silica gel
(1:2 EtOAc–petroleum ether) to give 12 (697 mg, 70%) as a white
0.20 mmol) in acetone (4.5 mL) and H2O (0.5 mL) at ambient tem-
perature was added NBS (110 mg, 0.60 mmol). After 10 min, the
reaction was quenched with satd aq NaHCO3 and the mixture con-
centrated. The crude intermediate was diluted with CHCl3 (15 mL)
and washed sequentially with satd aq NaHCO3 (10 mL), brine
(2 ꢂ 10 mL), dried over Na2SO4 and concentrated. The residue
was purified by silica gel column chromatography to afford a white
solid (82.0 mg). The white solid was added CCl3CN (0.11 mL,
solid; ½a 2D2
ꢃ
+10.2 (c 1.5, CHCl3); 1H NMR (600 MHz, CDCl3): d
7.07–7.79 (m, 18H, Ar), 4.66–4.81 (m, 4H, 2 ꢂ PhCH2), 4.50 (d,
1H, J 9.9 Hz, H-1), 4.25 (dd, 1H, J 10.4, 6.1 Hz, H-6), 4.19 (dd, 1H,
J 10.4, 7.1 Hz, H-60), 3.98 (d, 1H, J 3.3 Hz, H-4), 3.71 (t, 1H, J
9.3 Hz, H-2), 3.62–3.65 (m, 1H, H-5), 3.54 (dd, 1H, J 8.8, 3.3 Hz,
H-3), 2.41 (s, 3H, ArCH3), 2.31 (s, 3H, ArCH3); 13C NMR (150 MHz,
CDCl3): d 21.3, 21.9, 66.3, 68.5, 72.6, 75.4, 75.9, 76.9, 82.2, 88.1,
128.1–128.8, 129.8–130.1 (resonance overlap), 132.7, 132.8,
137.6, 138.0, 138.3, 145.2; LR-ESI-MS m/z calcd for [M+Na]+
643.2; found 643.1.
1.1 mmol) and DBU (13.7 lL, 0.092 mmol) in CH2Cl2 (4 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 8b (92 mg, 72%) as a syrup.
To a solution of trichloroacetimidate 8b (50.0 mg, 0.081 mmol)
and (S)-1,2-isopropylideneglycerol (13
(4 mL) was added freshly dried powdered 4 Å MS and the mixture
was cooled to 0 °C. TMSOTf (3.3 L, 0.016 mmol) was added to the
lL, 0.097 mmol) in dry Et2O
l
solution, and the mixture was stirred at 0 °C for 0.5 h. The mixture
was diluted with EtOAc (40 mL) and filtered through Celite. The or-
ganic layer was washed sequentially with satd aq NaHCO3 and
brine, dried (Na2SO4), and concentrated. The residue was purified
by column chromatography on silica gel (8:1 petroleum ether–
4.2.9. p-Tolyl-6-azido-2,3-di-O-benzyl-6-deoxy-1-thio-b-D-
galactopyranoside (13)
To a solution of 12 (500 mg, 0.80 mmol) in dry DMF (12 mL),
were added 15-crown-5 (0.16 mL, 0.80 mmol) and NaN3
(200 mg, 3.2 mmol). The reaction mixture was stirred at 65 °C
for 8 h, then cooled to rt, and water (20 mL) and CH2Cl2 (80 mL)
were added. The organic layer was dried over Na2SO4, filtered
and concentrated. The residue was purified on silica gel (1:4
EtOAc–petroleum ether) to give 13 (300 mg, 76%) as a thick oil;
EtOAc) to furnish 9
a
and 9b (43.6 mg, 86%,
a
/b = 2:5) as a syrup;
1H NMR (CDCl3, 600 MHz): d H-1 for 9
for 9b: 4.40 (d, J 7.7 Hz).
a: 4.93 (d, J 3.8 Hz), d H-1
4.2.6. p-Tolyl-4,6-O-Benzylidene-2,3-di-O-benzyl-1-thio-b-D-
galactopyranoside (10)31
½
a 2D2
ꢃ
+26.0 (c 4.0, CHCl3); IR (film, cmꢁ1): 3383.5 (
m
O–H), 3030.0
Compound 3 (0.5 g, 1.7 mmol) was dissolved in dry DMF (5 mL),
then benzaldehyde dimethyl acetal (0.3 mL, 2.0 mmol) and TsOH
(65 mg, 0.34 mmol) were added. After stirring at 50 °C for 6 h,
the mixture was neutralized by the addition of Et3N, and then
evaporated to dryness to provide a foamy solid. The crude foamy
solid (0.60 g, about 1.3 mmol) was dissolved in DMF (5 mL) and
then cooled to 0 °C. Sodium hydride (156 mg, 60%, 3.9 mmol)
was added, followed by BnBr (0.4 mL, 3.9 mmol). The reaction
was monitored by TLC and was complete after 2 h at 0 °C. The mix-
ture was diluted with water (15 mL) and extracted with CH2Cl2
(mAr-H), 2106.6
(
mN N), 1092.9 cmꢁ1 C–O); 1H NMR (600 MHz,
(m
„
CDCl3): d 7.10–7.49 (m, 14H, Ar), 4.86 (d, 1H, J 9.9 Hz, PhCH2),
4.74 (d, 1H, J 9.9 Hz, PhCH2), 4.69 (br s, 2H, PhCH2, H-1), 4.55
(d, 1H, J 9.9 Hz, PhCH2), 3.94 (br s, 1H, H-4, J 4.4 Hz), 3.65–3.70
(m, 2H, H-2, H-6), 3.57(dd, 1H, J 8.8, 3.3 Hz, H-3), 3.47–3.49 (m,
1H, H-5), 3.39 (dd, 1H, J 13.2, 5.5 Hz, H-60), 2.33 (s, 3H, ArCH3);
13C NMR (150 MHz, CDCl3): d 21.4, 51.3, 67.1, 72.7, 76.0, 76.9,
77.0, 82.5, 88.6, 128.1–128.8, 129.7–129.9 (resonance overlap),
133.1, 137.6, 138.2, 138.3; LR-ESI-MS m/z calcd for [M+Na]+
514.2; found 514.2.

20
C. Li et al. / Carbohydrate Research 376 (2013) 15–23
4.2.10. p-Tolyl-4-O-acetyl-6-azido-2,3-di-O-benzyl-6-deoxy-1-
thio-b- -galactopyranoside (14)
Na2SO4 and concentrated. The residue was chromatographed on
D
silica gel (1:8 EtOAc–petroleum ether) to give 16 (0.98 g, 82% over
To a solution of 13 (1.0 g, 2.0 mmol) in CH2Cl2 (15 mL), were
added Ac2O (0.38 mL, 4.0 mmol) and Et3N (0.55 mL, 4.0 mmol)
and catalytic DMAP (24 mg, 0.2 mmol). The reaction mixture was
stirred at room temperature for 0.5 h, then washed sequentially
with aq HCl (1 M) and brine. The organic phase was dried and con-
centrated and the residue was purified by column chromatography
two steps) as a thick oil. ½a D22
ꢃ
+20.6 (c 1.9, CHCl3); 1H NMR
(600 MHz, CDCl3): d 7.27–7.40 (m, 15H, Ar), 4.94 (d, 1H, J
3.3 Hz, H-1), 4.58–5.01 (m, 6H, 3 ꢂ PhCH2), 4.33–4.37 (m, 1H,
Hsn-2), 4.01–4.07 (m, 2H, H-3, H-5), 3.91 (dd, 1H, J 9.9, 3.3 Hz,
H-2), 3.84 (dd, 1H, J 8.8, 5.5 Hz, Hsn-3), 3.79 (d, 1H, J 2.2 Hz, H-
0
4), 3.74 (dd, 1H, J 8.8, 6.6 Hz, Hsn-3 ), 3.65 (dd, 1H, J 11.0, 5.5 Hz,
on silica gel to provide 14 (0.99 g, 92%) as a white solid; ½a D22
ꢃ
+3.7 (c
H
sn-1), 3.58 (dd, 1H, J 11.0, 5.5 Hz, Hsn-1 ), 3.49 (dd, 1H, J 12.1,
0
1.4, CHCl3); IR (film, cmꢁ1): 3027.1 (
m
Ar-H), 2103.7 (mN N), 1743.9
7.7 Hz, H-6), 2.96 (dd, 1H, J 12.1, 4.4 Hz, H-60), 1.40 (s, 3H, –
CH3), 1.37 (s, 3H, –CH3); 13C NMR (150 MHz, CDCl3): d 25.8,
27.0, 51.6, 66.9, 69.2, 70.2, 73.5, 73.8, 74.9 (2C), 75.3, 76.6, 98.2,
109.7, 127.8–128.1, 128.6–128.8 (resonance overlap), 138.3,
138.7, 138.9; LR-ESI-MS m/z calcd for [M+Na]+ 612.3, found
(mC O), 1230.3, 1102.3 (
m
C–O–C); 1H NMR (600 MHz,„ CDCl3): d
7.1@0–7.49 (m, 14H, Ar), 5.46 (d, 1H, J 4.4 Hz, H-4), 4.81 (d, 1H, J
9.8 Hz, PhCH2), 4.75 (d, 1H, J 9.8 Hz, PhCH2), 4.72 (d, 1H, J
11.0 Hz, PhCH2), 4.61 (d, 1H, J 7.7 Hz, H-1), 4.60 (d, 1H, J 11.0 Hz,
PhCH2), 3.61–3.64 (m, 3H, H-2, H-3, H-5), 3.53 (dd, 1H, J 12.1,
7.7 Hz, H-6), 3.22 (dd, 1H, J 12.1, 4.4 Hz, H-60), 2.33 (s, 3H, ArCH3),
2.17 (s, 3H, –COCH3); 13C NMR (150 MHz, CDCl3): d 21.1, 21.3, 51.4,
67.5, 72.3, 76.0, 76.9, 77.1, 81.1, 88.7, 128.1–128.7, 129.6–129.9
(resonance overlap), 133.2, 137.6, 138.2, 138.4, 170.1; LR-ESI-MS
m/z calcd for [M+Na]+ 556.2; found 556.1.
612.4. HR-ESI-MS m/z calcd for
C33H39O7N3 Na 612.2680
[M+Na]+, found 612.2684.
4.2.13. 3-O-(60-Amino-20,30,40-tri-O-benzyl-60-deoxy-
galactosyl)-sn-glycerol (17)
a-D-
TsOH (320 mg, 1.7 mmol) was added to a stirred solution of 16
(500 mg, 0.85 mmol) in MeOH (20 mL). After stirring for 2 h at
ambient temperature, the solution was concentrated and dissolved
in CH2Cl2. The organic layer was washed sequentially with satd aq
NaHCO3 and water, dried over Na2SO4, filtered and concentrated to
get a crude product. To a solution of the crude product in THF
(25 mL) and water (0.2 mL) was added PPh3 (450 mg, 3.4 mmol).
The reaction mixture was stirred at 50 °C for 5 h. The mixture
was evaporated under reduced pressure, and the residue was puri-
fied by silica gel column chromatography (CH2Cl2/MeOH) to give
17 (390 mg, 88% over two steps) as a colorless waxy substance;
4.2.11. 3-O-(40-O-Acetyl-60-azido-20,30-di-O-benzyl-60-deoxy-
galactosyl)-1,2-isopropylidene-sn-glycerol (15
a-D-
a)
A mixture of the glycosyl donor 14 (2.0 g, 3.8 mmol) and freshly
dried 4 Å molecular sieves was stirred in dry Et2O (150 mL) at room
temperature under a N2 atmosphere. The reaction mixture was
cooled in an ice bath and NIS (1.28 g, 5.7 mmol) was added. After
stirring for 10 min, DTBMP (780 mg, 3.8 mmol) was added, fol-
lowed by the addition of acceptor (S)-1,2-isopropylideneglycerol
(0.57 mL, 4.6 mmol) and TMSOTf (0.4 mL, 2.3 mmol). The reaction
mixture was stirred at 0 °C under a N2 atmosphere for 12 h, when
TLC showed that the reaction was complete. The mixture was
quenched by the addition of Et3N, filtered through a pad of Celite.
The filtrate was removed under reduced pressure and the residue
was dissolved in CH2Cl2. The solution was washed with 10%
Na2S2O3, followed by satd aq NaHCO3. The organic layer was dried
over Na2SO4, then the solvent was removed in vacuo, and the res-
idue was purified by column chromatography on silica gel to afford
½
a 2D2 +27.0 (c 1.2, CHCl3); 1H NMR (CDCl3, 600 MHz): d 7.26–7.40
ꢃ
(m, 15H, Ar), 4.59–4.95 (m, 6H, 3 ꢂ PhCH2), 4.77 (d, 1H, J 3.3 Hz,
H-1), 4.04 (dd, 1H, J 9.9, 4.4 Hz, H-3), 3.86–3.91 (m, 2H, H-5, Hsn-
0
2), 3.77–3.81 (m, 2H, Hsn-3, Hsn-3 ), 3.68 (dd, 1H, J 9.9, 5.5 Hz, Hsn-
0
1), 3.61 (d, 1H, J 4.4 Hz, H-4), 3.50 (dd, 1H, J 11.0, 6.6 Hz, Hsn-1 ),
3.05–3.10 (m, 3H, H-2, –NH2), 2.95 (dd, 1H, J 12.1, 8.8 Hz, H-6),
2.42 (dd, 1H, J 12.1, 3.3 Hz, H-60); LR-ESI-MS m/z calcd for
[M+H]+ 524.3; found 524.0. HR-ESI-MS m/z calcd for C30H38O7N
H 524.2643 [M+H]+, found 524.2649.
15
a
and 15b (1.82 g, 90%,
a
/b = 32:1) as a thick oil; for 15
a
: ½a 2D2
ꢃ
+89.0 (c 2.5, CHCl3); 1H NMR (600 MHz, CDCl3): d 7.27–7.36 (m,
10H, Ar), 5.46 (d, 1H, J 2.8 Hz, H-4), 4.97 (d, 1H, J 3.8 Hz, H-1),
4.82 (d, 1H, J 12.1 Hz, PhCH2), 4.70 (d, 1H, J 11.5 Hz, PhCH2), 4.64
(d, 1H, J 11.6 Hz, PhCH2), 4.57 (d, 1H, J 11.5 Hz, PhCH2), 4.33–
4.38 (m, 1H, Hsn-2), 4.03–4.09 (m, 2H, Hsn-3, H-5), 3.94 (dd, 1H, J
9.9, 3.3 Hz, H-3), 3.78 (dd, 1H, J 10.4, 3.8 Hz, H-2), 3.75 (dd, 1H, J
4.2.14. General procedure for 18a–g formation
To a solution of compound 17 (80 mg, 0.15 mmol) in dry pyri-
dine (4 mL), catalytic DMAP (2 mg, 0.015 mmol) 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 Na2SO4, filtered and concentrated. Purification by flash
chromatography (petroleum ether–EtOAc) yielded the correspond-
ing compound 18a–g (67–90%) as a colorless syrup or white solid.
0
8.2, 6.0 Hz, Hsn-3 ), 3.69 (dd, 1H, J 11.0, 5.0 Hz, Hsn-1), 3.60 (dd,
0
1H, J 10.4, 5.5 Hz, Hsn-1 ), 3.40 (dd, 1H, J 13.2, 8.2 Hz, H-6), 3.13
(dd, 1H, J 12.6, 4.4 Hz, H-60), 2.14 (s, 3H, –COCH3), 1.40 (s, 3H, –
CH3), 1.36 (s, 3H, –CH3); 13C NMR (150 MHz, CDCl3): d 21.1, 25.8,
27.0, 51.3, 66.8, 68.6, 68.8, 69.2, 72.5, 73.6, 74.9, 75.6, 75.8, 98.2,
109.8, 127.9–128.6 (resonance overlap), 138.1, 138.6, 170.6; LR-
ESI-MS m/z calcd for [M+Na]+ 564.6, found 564.0; HR-ESI-MS m/z
calcd for C28H35O8N3 Na 564.2316 [M+Na]+, found 564.2325.
4.2.14.1. 1,2-Dihexanoyl-3-(N-hexanoyl-60-amino-20,30,40-tri-O-
benzyl-60-deoxy-
78%).
d 7.26–7.40 (m, 15H, Ar), 5.37 (dd, 1H, J 6.6, 3.8 Hz, –NHCO–),
5.22–5.24 (m, 1H, Hsn-2), 4.63–4.99 (m, 6H, 3 ꢂ PhCH2), 4.80 (d,
1H, J 3.3 Hz, H-1), 4.35 (dd, 1H, J 12.1, 4.4 Hz, Hsn-1), 4.16 (dd, 1H,
a
-
D
-galactosyl)-sn-glycerol
(18a,
½ ꢃ
a 2D2
+23.8 (c 0.35, CHCl3); 1H NMR (CDCl3, 600 MHz):
4.2.12. 3-O-(60-Azido-20,30,40-tri-O-benzyl-60-deoxy-
galactosyl)-1,2-isopropylidene-sn-glycerol (16)
a-D-
To a solution of 15a (1.1 g, 2.0 mmol) in MeOH (30 mL) was
0
J 12.1, 6.6 Hz, Hsn-1 ), 4.02 (dd, 1H, J 9.9, 3.3 Hz, H-3), 3.89 (dd,
added catalytic NaOMe (11 mg, 0.2 mmol). The reaction mixture
was stirred for 30 min, and then was neutralized with Amberlite
IR120 resin (H+). After filtration, the filtrate was concentrated in
vacuo to get a residue. To a solution of the residue in DMF
(10 mL) was added BnBr (0.36 mL, 3.0 mmol). The reaction solu-
tion was cooled to 0 °C, and then NaH (160 mg, 4 mmol) was
slowly added in 10 min. After stirring for 2 h, the reaction mix-
ture was diluted with water (30 mL) and extracted with CH2Cl2
(30 mL ꢂ 3). All of the organic extracts were combined, dried over
1H, J 9.9, 2.2 Hz, H-2), 3.80 (br s, 1H, H-4), 3.73–3.75 (m, 1H, H-
5), 3.66 (dd, 1H, J 11.0, 5.5 Hz, Hsn-3), 3.56 (dd, 1H, J 11.0, 5.5 Hz,
0
0
H
sn-3 ), 3.38–3.42 (m, 1H, H-6), 3.19–3.24 (m, 1H, H-6 ), 2.24–2.32
(m, 4H, 2 ꢂ –CH2–COO–), 1.95 (t, 2H, J 7.7 Hz, –NHCO–CH2–),
1.48–1.65 (m, 6H, 3 ꢂ –CO–CH2–CH2–), 1.22–1.32 (m, 12H, 3 ꢂ –
CH2–CH2(CH2)2–CH3), 0.87–0.89 (m, 9H, 3 ꢂ –CH3); HR-ESI-MS
m/z calcd for
C48H68O10N N Na
818.4838 [M+H]+, C48H67O10
840.4657 [M+Na]+, found 818.4854 [M+H]+, 840.4671 [M+Na]+.

C. Li et al. / Carbohydrate Research 376 (2013) 15–23
21
4.2.14.2. 1,2-Dioctanoyl-3-(N-octanoyl-60-amino-20,30,40-tri-O-
benzyl-60-deoxy-
-galactosyl)-sn-glycerol (18b,
87%).
+20.8 (c 0.37, CHCl3); 1H NMR (CDCl3, 600 MHz):
d 7.26–7.40 (m, 15H, Ar), 5.37 (dd, 1H, J 7.2, 4.2 Hz, –NHCO–),
5.22–5.24 (m, 1H, Hsn-2), 4.63–4.98 (m, 6H, 3 ꢂ PhCH2), 4.80 (d,
1H, J 3.3 Hz, H-1), 4.35 (dd, 1H, J 12.1, 3.3 Hz, Hsn-1), 4.16 (dd, 1H,
4.2.14.6.
benzyl-60-deoxy-
67%).
+5.4 (c 0.10, CHCl3); 1H NMR (CDCl3, 600 MHz): d
7.26–7.40 (m, 15H, Ar), 5.36 (dd, 1H, J 7.7, 4.4 Hz, –NHCO–),
5.20–5.23 (m, 1H, Hsn-2), 4.62–4.98 (m, 6H, 3 ꢂ PhCH2), 4.80 (d,
1H, J 3.3 Hz, H-1), 4.34 (dd, 1H, J 12.1, 3.3 Hz, Hsn-1), 4.16 (dd, 1H,
1,2-Distearoyl-3-(N-stearoyl-60-amino-20,30,40-tri-O-
-galactosyl)-sn-glycerol (18f,
½ ꢃ
a-D
a-D
½
a 2D2
ꢃ
a 2D2
0
0
J 12.1, 6.6 Hz, Hsn-1 ), 4.02 (dd, 1H, J 9.9, 4.4 Hz, H-3), 3.89 (dd,
J 12.1, 6.6 Hz, Hsn-1 ), 4.02 (dd, 1H, J 9.9, 4.4 Hz, H-3), 3.88 (dd,
1H, J 9.9, 3.3 Hz, H-2), 3.79 (br s, 1H, H-4), 3.73–3.75 (m, 1H, H-
5), 3.65 (dd, 1H, J 11.0, 5.5 Hz, Hsn-3), 3.56 (dd, 1H, J 11.0, 5.5 Hz,
1H, J 9.9, 2.2 Hz, H-2), 3.80 (br s, 1H, H-4), 3.73–3.75 (m, 1H, H-
5), 3.65 (dd, 1H, J 11.0, 5.5 Hz, Hsn-3), 3.56 (dd, 1H, J 11.0, 6.6 Hz,
0
0
0
0
H
sn-3 ), 3.38–3.42 (m, 1H, H-6), 3.21–3.24 (m, 1H, H-6 ), 2.26–2.34
Hsn-3 ), 3.38–3.42 (m, 1H, H-6), 3.19–3.23 (m, 1H, H-6 ), 2.26–2.34
(m, 4H, 2 ꢂ –CH2–COO–), 1.95 (t, 2H, –NHCO–CH2–, J 7.7 Hz),
1.48–1.62 (m, 6H, 3 ꢂ –CO–CH2–CH2–), 1.20–1.30 (m, 24H, 3 ꢂ –
CH2–CH2(CH2)4–CH3), 0.86–0.88 (m, 9H, 3 ꢂ –CH3); LR-ESI-MS m/
(m, 4H, 2 ꢂ –CH2–COO–), 1.94 (t, 2H, J 7.7 Hz, –NHCO–CH2–),
1.49–1.62 (m, 6H, 3 ꢂ –CO–CH2–CH2–), 1.18–1.31 (m, 84H, 3 ꢂ –
CH2–CH2(CH2)14–CH3), 0.87 (t, 9H, J 6.6 Hz, 3 ꢂ –CH3); LR-ESI-MS
m/z calcd for [M+Na]+ 1345.0, found 1345.0.
z
924.6 [M+Na]+; HR-ESI-MS m/z calcd for C54H79O10
N Na
924.5596 [M+Na]+, found 924.5605.
4.2.14.7. 1,2-Diisovaleryl-3-(N-isovaleryl-60-amino-20,30,40-tri-O-
4.2.14.3. 1,2-Dilauroyl-3-(N-lauroyl-60-amino-20,30,40-tri-O-ben-
benzyl-60-deoxy-
82%).
d 7.26–7.40 (m, 15H, Ar), 5.35 (dd, 1H, J 7.2, 4.2 Hz, –NHCO–),
5.21–5.24 (m, 1H, Hsn-2), 4.63–4.99 (m, 6H, 3 ꢂ PhCH2), 4.79 (d,
1H, J 2.8 Hz, H-1), 4.35 (dd, 1H, J 12.1, 3.3 Hz, Hsn-1), 4.15 (dd, 1H,
a
-
D
-galactosyl)-sn-glycerol
(18g,
zyl-60-deoxy-
a
-
D
-galactosyl)-sn-glycerol (18c, 90%).
½
a 2D2
ꢃ
½ ꢃ
a 2D2
+23.8 (c 0.65, CHCl3); 1H NMR (CDCl3, 600 MHz):
+10.6 (c 0.20, CHCl3); 1H NMR (CDCl3, 600 MHz): d 7.27–7.40 (m,
15H, Ar), 5.35 (dd, 1H, J 7.1, 4.4 Hz, –NHCO–), 5.22–5.23 (m, 1H,
H
sn-2), 4.63–4.98 (m, 6H, 3 ꢂ PhCH2), 4.80 (d, 1H, J 3.3 Hz, H-1),
0
4.35 (dd, 1H, J 12.1, 3.3 Hz, Hsn-1), 4.16 (dd, 1H, J 12.1, 6.6 Hz,
J 12.1, 6.6 Hz, Hsn-1 ), 4.02 (dd, 1H, J 9.9, 3.3 Hz, H-3), 3.88 (dd,
0
Hsn-1 ), 4.02 (dd, 1H, J 9.9, 3.3 Hz, H-3), 3.88 (dd, 1H, J 9.9, 3.3 Hz,
1H, J 9.9, 2.2 Hz, H-2), 3.79 (br s, 1H, H-4), 3.72–3.74 (m, 1H, H-
5), 3.66 (dd, 1H, J 11.0, 5.5 Hz, Hsn-3), 3.57 (dd, 1H, J 11.0, 6.6 Hz,
H-2), 3.80–3.81 (br s, 1H, H-4), 3.73–3.75 (m, 1H, H-5), 3.65 (dd,
1H, J 11.0, 5.5 Hz, Hsn-3), 3.55 (dd, 1H, J 11.0, 5.5 Hz, Hsn-3 ), 3.38–
0
0
0
Hsn-3 ), 3.40–3.44 (m, 1H, H-6), 3.20–3.24 (m, 1H, H-6 ), 2.16–2.22
3.43 (m, 1H, H-6), 3.19–3.24 (m, 1H, H-60), 2.26–2.34 (m, 4H,
2 ꢂ –CH2–COO–), 1.95 (t, 2H, –NHCO–CH2–, J 6.6 Hz), 1.48–1.65
(m, 6H, 3 ꢂ –CO–CH2–CH2–), 1.20–1.31 (m, 48H, 3 ꢂ –CH2–
CH2(CH2)8–CH3), 0.88 (t, 9H, J 6.6 Hz, 3 ꢂ –CH3); HR-ESI-MS m/z
calcd for C66H103O10N Na 1092.7474 [M+Na]+, found 1092.7458.
(m, 4H, 2 ꢂ –CH2–COO–), 1.81–2.09 (m, 5H, –NHCO–CH2–, 3 ꢂ –
CH(CH3)2), 0.85–0.95 (m, 18H, 6 ꢂ –CH3). LR-ESI-MS m/z 776.4
[M+H]+, 798.4 [M+Na]+; HR-ESI-MS m/z calcd for C45H62O10
N
776.4368 [M+H],
C45H61O10N Na 798.4188 [M+Na], found
776.4375 [M+H]+, 798.4194 [M+Na]+.
4.2.15. General procedure for 2a–g formation
4.2.14.4. 1,2-Dimyristoyl-3-(N-myristoyl-60-amino-20,30,40-tri-O-
A solution of 18a–g (100 mg) in THF/i-PrOH (9:1, 20 mL) was
treated with 10% palladium(II) hydroxide (100 mg) and stirred at
ambient temperature under a hydrogen atmosphere for 8 h. After
filtration, the solvent was evaporated and the residue was purified
by column chromatography (CH2Cl2–MeOH) to afford 2a–g (82–
90%) as a colorless syrup or waxy solid.
benzyl-60-deoxy-
83%).
a
-
D
-galactosyl)-sn-glycerol
(18d,
½ ꢃ
a 2D2
+10.8 (c 0.35, CHCl3); 1H NMR (CDCl3, 600 MHz):
d 7.26–7.40 (m, 15H, Ar), 5.36 (dd, 1H, J 7.1, 3.8 Hz, –NHCO–),
5.22–5.23 (m, 1H, Hsn-2), 4.62–4.98 (m, 6H, 3 ꢂ PhCH2), 4.80 (d,
1H, J 3.3 Hz, H-1), 4.35 (dd, 1H, J 12.1, 3.3 Hz, Hsn-1), 4.16 (dd, 1H,
0
J 12.1, 5.5 Hz, Hsn-1 ), 4.02 (dd, 1H, J 9.9, 3.3 Hz, H-3), 3.88 (dd,
1H, J 9.9, 3.3 Hz, H-2), 3.80 (br s, 1H, H-4), 3.73–3.75 (m, 1H, H-
5), 3.65 (dd, 1H, J 11.0, 5.5 Hz, Hsn-3), 3.55 (dd, 1H, J 11.0, 5.5 Hz,
4.2.15.1. 1,2-Dihexanoyl-3-(N-hexanoyl-60-amino-60-deoxy-
a-D-
galactosyl)-sn-glycerol (2a, 82%).
½
a 2D2
+109.2 (c 0.5, CHCl3);
ꢃ
0
0
H
sn-3 ), 3.38–3.41 (m, 1H, H-6), 3.21–3.23 (m, 1H, H-6 ), 2.26–2.35
1H NMR (CDCl3, 600 MHz): d 6.46 (dd, 1H, J 12.1, 6.6 Hz, –NHCO–),
5.24–5.26 (m, 1H, Hsn-2), 4.88 (d, 1H, J 3.3 Hz, H-1), 4.35 (dd, 1H, J
(m, 4H, 2 ꢂ –CH2–COO–), 1.94 (t, 2H, J 7.7 Hz, –NHCO–CH2–),
1.47–1.63 (m, 6H, 3 ꢂ –CO–CH2–CH2–), 1.18–1.31 (m, 60H, 3 ꢂ –
CH2–CH2(CH2)10–CH3), 0.87 (t, 9H, J 7.6 Hz, 3 ꢂ –CH3); LR-ESI-MS
0
12.1, 3.3 Hz, Hsn-1), 4.14 (dd, 1H, J 12.1, 6.6 Hz, Hsn-1 ), 3.66–3.90
0
(m, 7H, H-2, H-3, H-4, H-5, H-6, Hsn-3, Hsn-3 ), 3.26–3.28 (m, 1H,
m/z 1176.9 [M+Na]+; HR-ESI-MS m/z calcd for C72H116O10
N
H-60), 2.30–2.34 (m, 4H, 2 ꢂ –CH2–COO–), 2.20 (t, 2H, J 7.7 Hz, –
NHCO–CH2–), 1.59–1.62 (m, 6H, 3 ꢂ –CO–CH2–CH2–), 1.25–1.34
(m, 12H, 3 ꢂ –CH2–CH2(CH2)2CH3), 0.86–0.90 (m, 9H, 3 ꢂ –CH3);
13C NMR (150 MHz, CDCl3): d 14.1 (3CH3), 22.5 (2CH2), 24.7, 24.8,
25.6, 29.9, 31.4, 31.6, 31.7, 34.3, 34.5, 36.7, 39.5 (C-60), 62.7, 67.5,
69.0, 69.1, 69.4, 70.3, 70.4, 100.1 (C-10), 173.7, 174.0, 175.2; LR-
ESI-MS m/z 548.2 [M+H]+, 570.2 [M+Na]+; HR-ESI-MS m/z calcd
1154.8594 [M+H]+, C72H115O10N Na 1176.8413 [M+Na]+, found
1154.8571 [M+H]+, 1176.8405 [M+Na]+.
4.2.14.5. 1,2-Dipalmitoyl-3-(N-palmitoyl-60-amino-20,30,40-tri-O-
benzyl-60-deoxy-
74%).
d 7.26–7.40 (m, 15H, Ar), 5.37 (dd, 1H, J 7.2, 4.2 Hz, –NHCO–),
5.21–5.23 (m, 1H, Hsn-2), 4.62–4.98 (m, 6H, 3 ꢂ PhCH2), 4.80 (d,
1H, J 3.3 Hz, H-1), 4.35 (dd, 1H, J 12.1, 3.3 Hz, Hsn-1), 4.16 (dd, 1H,
a
-
D
-galactosyl)-sn-glycerol
(18e,
½ ꢃ
a 2D2
+12.0 (c 0.60, CHCl3); 1H NMR (CDCl3, 600 MHz):
for
C27H50O10N N Na 570.3249
548.3429 [M+H]+, C27H49O10
[M+Na]+, found 548.3436 [M+H]+, 570.3254 [M+Na]+.
4.2.15.2. 1,2-Dioctanoyl-3-(N-octanoyl-60-amino-60-deoxy-
a-D-
0
J 12.1, 6.6 Hz, Hsn-1 ), 4.02 (dd, 1H, J 9.9, 4.4 Hz, H-3), 3.88 (dd,
galactosyl)-sn-glycerol (2b, 88%).
½
a 2D2
+69.7 (c 1.4, CHCl3);
ꢃ
1H, J 9.9, 3.3 Hz, H-2), 3.80 (br s, 1H, H-4), 3.73–3.75 (m, 1H, H-
1H NMR (CDCl3, 600 MHz): d 6.29 (dd, 1H, –NHCO–, J 12.1,
6.6 Hz), 5.24–5.26 (m, 1H, Hsn-2), 4.88 (d, 1H, J 3.3 Hz, H-1), 4.33
5), 3.65 (dd, 1H, J 11.0, 5.5 Hz, Hsn-3), 3.55 (dd, 1H, J 11.0, 5.5 Hz,
0
0
Hsn-3 ), 3.38–3.42 (m, 1H, H-6), 3.19–3.23 (m, 1H, H-6 ), 2.25–2.35
0
(dd, 1H, J 12.1, 3.3 Hz, Hsn-1), 4.14 (dd, 1H, J 12.1, 6.6 Hz, Hsn-1 ),
(m, 4H, 2 ꢂ –CH2COO–), 1.95 (t, 2H, J 6.6 Hz, –NHCO–CH2–),
1.48–1.64 (m, 6H, 3 ꢂ –CO–CH2–CH2–), 1.20–1.31 (m, 72H, 3 ꢂ –
CH2–CH2(CH2)12–CH3), 0.87 (t, 9H, J 6.6 Hz, 3 ꢂ –CH3); LR-ESI-MS
0
3.66–3.84 (m, 7H, H-2, H-3, H-4, H-5, H-6, Hsn-3, Hsn-3 ), 3.20–
3.22 (m, 1H, H-60), 2.30–2.36 (m, 4H, 2 ꢂ –CH2–COO–), 2.20 (t,
2H, J 7.7 Hz, –NHCO–CH2–), 1.58–1.62 (m, 6H, 3 ꢂ –CO–CH2–
CH2–), 1.22–1.32 (m, 24H, 3 ꢂ –CH2–CH2(CH2)4CH3), 0.88 (t, 9H, J
6.6 Hz, 3 ꢂ –CH3); 13C NMR (150 MHz, CDCl3): d 14.3 (3CH3),
22.8, 25.1, 25.2, 25.9, 29.2–29.5 (resonance overlap), 31.9, 34.3,
m/z 1260.9 [M+Na]+; HR-ESI-MS m/z calcd for C78H128O10
N
1238.9533 [M+H]+, C78H127O10N Na 1260.9352 [M+Na]+, found
1238.9545 [M+H]+, 1260.9370 [M+Na]+.

22
C. Li et al. / Carbohydrate Research 376 (2013) 15–23
34.5, 36.8, 39.7 (C-60), 62.6, 67.4, 68.9, 69.3, 69.5, 70.3, 70.4, 100.1
(C-10), 173.7, 173.9, 175.0; LR-ESI-MS m/z 632.5 [M+H]+, 654.5
[M+Na]+; HR-ESI-MS m/z calcd for C33H62O10N 632.4368 [M+H]+,
100.1 (C-10), 173.6, 173.8, 175.1; HR-MALDI-MS m/z calcd for
C63H121O10NNa 1074.8888 [M+Na]+, found 1074.8886.
C
33H61O10
N
Na 654.4188 [M+Na]+, found 632.4376 [M+H]+,
4.2.15.7. 1,2-Diisovaleryl-3-(N-isovaleryl-60-amino-60-deoxy-
a-
654.4194 [M+Na]+.
D
-galactosyl)-sn-glycerol (2g, 83%).
½
a 2D2
+75.0 (c 2.2, CHCl3);
ꢃ
1H NMR (CDCl3, 600 MHz): d 6.57 (dd, 1H, J 12.1, 6.6 Hz, –NHCO–),
4.2.15.3.
1,2-Dilauroyl-3-(N-lauroyl-60-amino-60-deoxy-
a-D-
5.25–5.27 (m, 1H, Hsn-2), 4.88 (d, 1H, J 2.8 Hz, H-1), 4.36 (dd, 1H, J
a 2D2
ꢃ
+56.2 (c 1.9, CHCl3);
12.1, 2.2 Hz, Hsn-1), 4.14 (dd, 1H, J 11.5, 6.1 Hz, Hsn-1 ), 3.65–3.88
0
galactosyl)-sn-glycerol (2c, 90%).
½
1H NMR (CDCl3, 600 MHz): d 6.35 (dd, 1H, J 12.1, 6.6 Hz, –NHCO–
), 5.24–5.26 (m, 1H, Hsn-2), 4.87 (d, 1H, J 3.3 Hz, H-1), 4.34 (dd,
(m, 7H, H-2, H-3, H-4, H-5, H-6, Hsn-3, Hsn-3 ), 3.27–3.30 (m, 1H,
0
H-60), 2.17–2.22 (m, 4H, 2 ꢂ –CH2–COO–), 2.05–2.10 (m, 5H, –
NHCO–CH2–, 3 ꢂ –CH(CH3)2), 0.92–0.97 (m, 18H, 6 ꢂ –CH3); 13C
NMR (150 MHz, CDCl3): d 22.3–22.7 (resonace overlap), 25.8,
25.9, 26.2, 39.8 (C-60), 43.3, 43.5, 45.9, 62.7, 67.1, 69.2, 69.4, 70.1,
70.2, 70.3, 100.1 (C-10), 172.9, 173.2, 174.1; LR-ESI-MS m/z 506.2
0
1H, J 11.0, 3.3 Hz, Hsn-1), 4.14 (dd, 1H, J 12.1, 5.5 Hz, H sn-1 ),
0
3.65–3.85 (m, 7H, H-2, H-3, H-4, H-5, H-6, Hsn-3, Hsn-3 ), 3.21–
3.23 (m, 1H, H-60), 2.30–2.34 (m, 4H, 2 ꢂ –CH2–COO–), 2.19 (t,
2H, J 7.4 Hz, –NHCO–CH2–), 1.59–1.62 (m, 6H, 3 ꢂ –CO–CH2–
CH2–), 1.22–1.32 (m, 48H, 3 ꢂ –CH2–CH2(CH2)8CH3), 0.88 (t, 9H, J
6.6 Hz, 3 ꢂ –CH3); 13C NMR13C NMR (150 MHz, CDCl3): d 14.3
(3CH3), 22.9, 25.1, 25.2, 25.9, 29.4–30.0 (resonance overlap), 32.1,
34.3, 34.5, 36.8, 39.7 (C-60), 62.7, 67.3, 69.1, 69.3, 69.4, 70.3, 70.4,
100.1 (C-10), 173.6, 173.9, 175.0; LR-ESI-MS m/z 800.4 [M+H]+;
HR-ESI-MS m/z calcd for C45H86O10N 800.6246 [M+H]+, found
800.6253.
[M+H]+, 528.2 [M+Na]+; HR-ESI-MS m/z calcd for C24H44O10
N
506.2960 [M+H]+, found 506.2959.
4.3. Bioassay methods
The Myt1 assay was performed using the ADPGlo™ assay kit
(Promega) which measures the generation of ADP by Myt1. Gener-
ation of ADP by the Myt1 reaction leads to an increase in the lumi-
nescence signal in the presence of™ assay kit.
4.2.15.4. 1,2-Dimyristoyl-3-(N-myristoyl-60-amino-60-deoxy-
a-
D
–
a
-
D
-galactosyl)-sn-glycerol (2d, 85%).
½
a 2D2
ꢃ
+46.0 (c 1.0,
The Myt1 assay was performed at 30 °C for 30 min in a final vol-
CHCl3); 1H NMR (CDCl3, 600 MHz): d 6.08 (dd, 1H, J 12.1, 6.6 Hz,
–NHCO–), 5.25–5.27 (m, 1H, Hsn-2), 4.88 (d, 1H, J 3.3 Hz, H-1),
4.32 (dd, 1H, J 12.1, 3.8 Hz, Hsn-1), 4.13 (dd, 1H, J 12.1, 6.1 Hz,
ume of 25
of diluted active Myt1, 5
L kinase assay buffer, 5
of 50 mM ATP stock solution.
l
L according to the following assay reaction recipe: 5
L of stock solution of peptide substrate,
L of compound or 10% DMSO, and 5
lL
l
l
5
l
lL
0
0
H
sn-1 ), 3.67–3.81 (m, 7H, H-2, H-3, H-4, H-5, H-6, Hsn-3, Hsn-3 ),
3.13–3.15 (m, 1H, H-60), 2.30–2.34 (m, 4H, 2 ꢂ –CH2–COO–), 2.20
(t, 2H, J 7.7 Hz –NHCO–CH2–), 1.59–1.62 (m, 6H, 3 ꢂ –CO–CH2–
CH2–), 1.21–1.31 (m, 60H, 3 ꢂ –CH2–CH2(CH2)10–CH3), 0.87 (t,
9H, J 7.1 Hz, 3 ꢂ –CH3); 13C NMR (150 MHz, CDCl3): d 14.4
(3CH3), 22.9, 25.1, 25.2, 25.9, 29.4–30.0 (resonance overlap), 32.1,
34.3, 34.5, 36.8, 39.6 (C-60), 62.5, 67.7, 68.7, 69.4, 69.8, 70.3, 70.4,
100.1 (C-10), 173.6, 173.9, 175.2; LR-ESI-MS m/z 884.8 [M+H]+;
HR-ESI-MS m/z calcd for C51H98O10N 884.7185 [M+H]+, found
884.7198.
The assay was started by incubating the reaction mixture in a
96-well plate at 30 °C for 30 min. After a 30 min’ incubation period,
the assay was terminated by the addition of 25 mL of ADPGlo™ re-
agent (Promega). The 96-well plate was shaken and then incubated
for 40 min at ambient temperature. Then 50 mL of Kinase Detec-
tion Reagent was added, the 96-well plate shaken and then incu-
bated for a further 30 min at ambient temperature. The 96-well
reaction plate was then read using the ADPGlo™ Luminescence
Protocol on a GloMax plate reader (Promega; Catalog #E7031).
Blank control was set up that included all the assay components
except the addition of the substrate (replace with equal volume of
kinase assay buffer). The corrected activity for Myt1 target was
determined by removing the blank control value.
4.2.15.5. 1,2-Dipalmitoyl-3-(N-palmitoyl-60-amino-60-deoxy-
a-
D
-galactosyl)-sn-glycerol (2e, 85%)17
.
½
a 2D2
+63.2 (c 1.0,
ꢃ
CHCl3); 1H NMR (CDCl3, 600 MHz): d 6.08 (dd, 1H, J 12.1, 6.6 Hz,
–NHCO–), 5.24–5.26 (m, 1H, Hsn-2), 4.87 (d, 1H, J 3.3 Hz, H-1),
4.32 (dd, 1H, J 12.1, 3.8 Hz, Hsn-1), 4.13 (dd, 1H, J 12.1, 6.1 Hz,
Acknowledgements
0
0
H
sn-1 ), 3.66–3.82 (m, 7H, H-2, H-3, H-4, H-5, H-6, Hsn-3, Hsn-3 ),
3.15–3.18 (m, 1H, H-60), 2.30–2.34 (m, 4H, 2 ꢂ –CH2–COO–), 2.20
(t, 2H, J 7.7 Hz, –NHCO–CH2–), 1.59–1.62 (m, 6H, 3 ꢂ –CO–CH2–
CH2–), 1.21–1.31 (m, 72H, 3 ꢂ –CH2–CH2(CH2)12–CH3), 0.87 (t,
9H, J 6.6 Hz, 3 ꢂ –CH3); 13C NMR (150 MHz, CDCl3): d 14.3
(3CH3), 22.9, 25.1, 25.2, 25.9, 29.6–30.0 (resonance overlap), 32.2,
34.3, 34.6, 36.8, 39.7 (C-60), 62.6, 67.7, 68.8, 69.4, 69.8, 70.4, 70.5,
100.2 (C-10), 173.6, 173.8, 175.1; HR-MALDI-MS m/z calcd for
This work was supported by the National Natural Science Foun-
dation of China (No. 81072574), program for New Century Excel-
lent Talents in University (NCET-08-0506), Special Fund for
Marine Scientific Research in the Public Interest (No. 201005024)
and PCSIRT (No. IRT09444).
Supplementary data
C
57H109O10NNa 990.7949 [M+Na]+, found 990.7919.
Supplementary data associated with this article can be found, in
the
online
version,
at
http://dx.doi.org/10.1016/
4.2.15.6.
1,2-Distearoyl-3-(N-stearoyl-60-amino-60-deoxy-
a-D-
j.carres.2013.02.008.
galactosyl)-sn-glycerol (2f, 82%).
½
a 2D2
+31.5 (c 2.6, CHCl3);
ꢃ
1H NMR (CDCl3, 600 MHz): d 6.09 (dd, 1H, J 11.0,5.5 Hz, –NHCO–
), 5.24–5.26 (m, 1H, Hsn-2), 4.88 (d, 1H, J 3.3 Hz, H-1), 4.32 (dd,
References
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1H, J 11.0, 3.3 Hz, Hsn-1), 4.13 (dd, 1H, J 12.1, 6.1 Hz, Hsn-1 ), 3.67–
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0
3.86 (m, 7H, H-2, H-3, H-4, H-5, H-6, Hsn-3, Hsn-3 ), 3.13–3.17 (m,
1H, H-60), 2.30–2.34 (m, 4H, 2 ꢂ –CH2–COO–), 2.19 (t, 2H, J
7.7 Hz, –NHCO–CH2–), 1.57–1.64 (m, 6H, 3 ꢂ –CO–CH2–CH2–),
1.15–1.35 (m, 84H, 3 ꢂ –CH2–CH2(CH2)14–CH3), 0.88 (t, 9H,
J
6.6 Hz, 3 ꢂ –CH3); 13C NMR (150 MHz, CDCl3): d 14.2 (3CH3),
22.9, 25.1, 25.2, 25.9, 29.6–29.9 (resonace overlap), 32.2, 34.3,
34.5, 36.8, 39.6 (C-60), 62.6, 67.6, 68.9, 69.3, 69.7, 70.3, 70.4,

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