Structure-activity relationship studies on acremomannolipin A, the potent calcium signal modulator with a novel glycolipid structure 3: Role of the length of alditol side chain

Article abstract of DOI:10.1016/j.bmc.2015.03.079

Abstract Five homologs of a novel glycolipid acremomannolipin A (1a), the potential Ca²⁺ signal modulator isolated from Acremonium strictum, bearing alditols of different length (1g-1k) were synthesized by a stereoselective β-mannosylation of a

Full text of DOI:10.1016/j.bmc.2015.03.079

Bioorganic & Medicinal Chemistry 23 (2015) 3761–3773
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Structure–activity relationship studies on acremomannolipin A,
the potent calcium signal modulator with a novel glycolipid
structure 3: Role of the length of alditol side chain
Nozomi Tsutsui a, Genzoh Tanabe a, Nao Morita a, Yoshitomo Okayama a, Ayako Kita b, Reiko Sugiura b,
,
⇑
Osamu Muraoka a,
⇑
a Laboratory of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Kinki University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan
b Laboratory of Molecular Pharmacogenomics, Faculty of Pharmacy, Kinki University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan
a r t i c l e i n f o
a b s t r a c t
Five homologs of a novel glycolipid acremomannolipin A (1a), the potential Ca2+ signal modulator isolated
from Acremonium strictum, bearing alditols of different length (1g–1k) were synthesized by a stereoselec-
tive b-mannosylation of appropriately protected mannosyl sulfoxide (2) with five alditols (1g: C2, 1h: C3,
1i: C4, 1j: C5 and 1k: C7 units), and their potential in modulating Ca2+ signaling were evaluated. Homologs
with alditols of more than 4 carbons (1i, 1j and 1k) were equally or more potent than the parent compound
(1a) regardless of the length of the alditol chain. Whereas activities of two homologs with shorter chains
(1g and 1h) decreased to a considerable extent. The results indicated that the length of the alditol side
chain was a crucial determinant for the potent calcium signal modulating activity.
Article history:
Received 5 February 2015
Revised 30 March 2015
Accepted 31 March 2015
Available online 9 April 2015
Keywords:
Acremomannolipin A
Glycolipid
Calcium signal modulator
Acremonium strictum
SAR study
Ó 2015 Elsevier Ltd. All rights reserved.
1. Introduction
Concurrently, its a-isomer (epi-1a) was also synthesized and
evaluated. The activity decreased upon inversion of the
configuration at the anomeric center, but remained to some
extents. Thus, the b-configuration was found not critical for onset
of the activity, but preferable for the activity4b (Fig. 1).
In our continuing study on the structure–activity relationship
(SAR) of 1a, we next synthesized five side chain diastereomers
(1b–1f) in order to clarify the role of stereochemistry of the alditol
side chain. Activities of all candidates were nearly equal or slightly
superior to that of 1a regardless of the stereochemistry of the aldi-
tol, revealing that the stereochemistry of the alditol moiety was
not critical for the potent calcium signal modulating activity4c
(Fig. 2). It was speculated that the hexitol chain would function
to control the hydrophilic property of the molecule.
Calcium signaling pathways are known to be widely pro-
liferated throughout the cell and participate in all basic cellular
functions.1 The ability to manipulate Ca2+ signaling pathways
would provide a powerful tool for applications in therapeutic
and biotechnological settings. Recently, we identified a novel
glycolipid acremomannolipin A2 (1a), a constituent isolated from
Acremonium strictum, as a potential calcium signal modulator using
a newly developed chemical-genetic method.3 The structure of 1a
is quite unique, in which D-mannopyranose is connected to D-man-
nitol through a b-glycoside linkage. All the hydroxyls in the man-
nose are highly masked by aliphatic acids to form peresters, and
thus the moiety is hydrophobic, whereas the mannitol part exhi-
bits the highly hydrophilic property.2 The compound (1a) was
found to show quite characteristic bioactive property, enabling cal-
cineurin deletion cells to grow in the presence of Clꢀ, which would
be caused by calcium signal modulating. The activity was so potent
as to exert the effect at a concentration of 200 nM. The first total
synthesis of this unique compound was accomplished by us
In the present study, five homologs with alditols of different
carbon chain length having possibly the same stereochemistry as
1a (1g–1k) were synthesized and their potential as calcium signal
modulators were examined (Fig. 3). Contribution of hydrophilicity
caused by the alditol side chain length to the activity is discussed.
via
a
stereoselective b-mannosylation as the key reaction.4a
2. Results and discussion
2.1. Preparation of acceptors (3g–3k) for mannosylation
⇑
Corresponding authors. Tel.: +81 6 4307 3642; fax: +81 6 6730 1394 (R.S.);
tel.: +81 6 4307 4015; fax: +81 6 6721 2505 (O.M.).
E-mail addresses: sugiurar@phar.kindai.ac.jp (R. Sugiura), muraoka@phar.kindai.
ac.jp (O. Muraoka).
In the synthetic methodology especially for the sugar related
molecules, selection of the protecting groups is the most important
http://dx.doi.org/10.1016/j.bmc.2015.03.079
0968-0896/Ó 2015 Elsevier Ltd. All rights reserved.

3762
N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
OCO(CH2)6CH3
with DMP to give 5-O-(tert-butyldiphenylsilyl)-1,2:3,4-di-O-iso-
propylidene- -arabinitol (15) in overall 40% yield via three steps
from 12. The perprotected arabinitol derivative 15 was then con-
verted to the target acceptor 3j in 68% yield through de-silylation
by using TBAF.
CH3(CH2)4COO
CH3(CH2)4COO
CH3(CH2)4COO
O
OH OH
D
O
OH
OH OH
Acremomannolipin A (1a)
The seven-carbon acceptor, 1,2:3,4:5,6-tri-O-isopropylidene-D-
OCO(CH2)6CH3
O
glycero-D-galacto-heptitol (3k), was prepared starting from 12
CH3(CH2)4COO
CH3(CH2)4COO
CH3(CH2)4COO
according to the protocol reported by us9 (Scheme 2).
OH OH
O
2.2. Syntheses of homologs of acremomannolipin A (1g–1k)
OH
epi-1a
OH OH
With 5 acceptors (3g–3k) in hand, the coupling reaction of
these 5 acceptors with mannosyl sulfoxide 2 in the presence of tri-
fluoromethanesulfonic anhydride (Tf2O) was examined. The cou-
pling reaction of 2 with all the acceptors 3g, 3h, 3i, 3j and 3k
Figure 1. Acremomannolipin A (1a) and its
a
-anomer (epi-1a).
step. In the total synthesis of 1a, 4,6-benzylidene protected man-
nosyl sulfoxide (2) was proved to be an appropriate donor for the
stereoselective b-mannosylation with mannitol as the acceptor.5
Therefore, the target glycolipids 1g–1k were synthesized by apply-
ing the b-selective mannosylation of the donor 2 with protected
alditols (3g–3k) as acceptors (Scheme 1).
showed excellent b-selectivity (b/a = >ca. 20/1) giving correspond-
ing b-mannosides 16g, 16h, 16i, 16j and 16k in 73, 40, 60, 52 and
42% yield, respectively (Scheme 3). The NMR spectral properties of
the products were similar with each other. In the 1H NMR spectra
of 16g–16k, signals due to anomeric protons were shown at d
4.50–4.57. The b-mannoside structure was confirmed by the NOE
correlations between the signal due to the anomeric proton at
C10 and signals due to C30-H and C50-H as depicted in Scheme 3.
The selective removal of the PMB group at C20 in the coupled prod-
ucts (16g–16k) were successfully conducted by DDQ oxidation to
give corresponding monoalcohols (17g–17k) in 79–92% yields,
which were then treated with octanoyl chloride to give the
corresponding octanoyl esters (18g–18k) in good yields.
Simultaneous hydrogenolysis of both the benzyl and benzylidene
moieties of 18g–18k over palladium on carbon gave the desired
trisalcohols (19g–19k), which were then derived to the
corresponding tetraesters (20g–20k), by treating with hexanoic
anhydride, in 63–86% yields in two steps. Finally, the acetonide
moieties and/or the TBDMS group in 20g–20k were selectively
removed without affecting ester moieties under acidic conditions
to give the target homologs (1g–1k) of acremomannolipin A (1a)
in 67–94% yields. The 13C NMR spectroscopic properties of man-
nose moiety in all the final products were similar with those of
1a as shown in Table 1.
Firstly the two-carbon acceptor, 2-(tert-butyldimethylsilylox-
y)ethanol (3g), was prepared by the mono-silylation of ethylene
glycol (4) according to the literature.6 The three carbon acceptor,
(R)-(ꢀ)-2,2-dimethyl-1,3-dioxolane-4-methanol (3h), was com-
mercially available. Isopropylidene-
as follows. According to the literature7,
verted to 2,4-O-benzylidene- -erythritol (6). The primary hydroxyl
of 6 was selectively protected with an acetyl (Ac) moiety to
give 1-O-acetyl-2,4-O-benzylidene- -erythritol (7) in 89% yield.
D
-erythritol (3i), was prepared
D-glucose (5) was con-
D
D
Subsequent regioselective reductive-cleavage of benzylidene
acetal 7 was achieved by treatment with triethylsilane in the pres-
ence of BF3ꢁEt2O to give 1-O-acetyl-4-O-benzyl-
which was acetalized with 2,2-dimethoxypropane (DMP) to give
1-O-acetyl-4-O-benzyl-2,3-O-isopropylidene- -erythritol (9). The
deacetylation of 9 and subsequent silylation of the resulting
monoalcohol, 4-O-benzyl-2,3-O-isopropylidene- -erythritol (10),
4-O-benzyl-1-O-(tert-butyldimethylsilyl)-2,3-O-iso-
D-erythritol (8),
D
D
gave
propylidene-D-erythritol (11). Hydrogenolysis of 11 gave the
desired acceptor 3i in 98% yield.
The five-carbon acceptor, 1,2:3,4-di-O-isopropylidene-D-ara-
binitol (3j), was synthesized as follows. The primary hydroxyl in
-arabinose (12) was selectively protected with the tert-
butyldiphenylsilyl (TBDPS) moiety to give a silyl ether, 5-O-(tert-
butyldiphenylsilyl)-
-arabinose8 (13). After the NaBH4 reduction
of the hemiacetal moiety in 13, four hydroxyls in the resulting tet-
2.3. Bioassay of acremomannolipin A homologs (1g–1k)
D
The potencies of the synthesized homologs 1g–1k as calcium
signal modulators were examined.
D
Disruption of the calcineurin gene (ppb1+) in fission yeast
resulted in a Clꢀ-sensitive growth defect, thus calcineurin gene
raol, 5-O-(tert-butyldiphenylsilyl)-
D
-arabinitol (14) were protected
deletion cells (Dppb1) can grew in yeast extract peptone dextrose
(YPD) medium, but fail to grow in YPD medium containing
0.11 M MgCl2 [see filter papers disc containing DMSO (control) in
Fig. 4]. When a solution of acremomannolipin A (1a) in DMSO
OH OH
OCO(CH2)6CH3
O
CH3(CH2)4COO
CH3(CH2)4COO
CH3(CH2)4COO
a : R =
OH
O
was added onto the filter paper disc,
Dppb1 cells grew well around
R
OH OH
Acremomannolipin A
OH OH
1
OCO(CH2)6CH3
O
CH3(CH2)4COO
CH3(CH2)4COO
CH3(CH2)4COO
OH OH
b : R =
d : R =
c : R =
O
OH
OH
OH
R
1
OH OH
OH OH
OH OH
OH
OH
4
2
OH OH
OH
g : R =
i : R =
h : R =
i : R =
OH
OH
1
OH
1
3
1
e : R =
OH
OH
OH OH
OH OH
OH OH
OH
OH OH OH
OH OH
OH
k : R =
f : R =
1
7
1
5
OH
OH OH
OH OH
Figure 3. Homologs 1g–1k of acremomannolipin A (1a) with different alditol chain
Figure 2. Acremomannolipin A (1a) and its side chain stereoisomers 1b–1f.
length.

N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
3763
OPMB
O
In summary, five homologs (1g–1k) of acremomannolipin A
(1a) with alditols of different carbon length from that of 1a were
synthesized by a stereoselective b-mannosylation of mannose 2
with five alditols, and their calcium signal modulating potency
were evaluated. Alditols composed of at least four carbons were
necessary for the compound to exert the potent activity. Studies
on the target of this molecule and further structure–activity
relationships of the characteristic activity of 1a as a potent calcium
signal modulator are in progress.
O
Ph
O
BnO
2
S
Ph
O
+
O
HO
O
OTBDMS
HO
3g
1g−1k
3h
O
HO
3. Experimental
3.1. General
OTBDMS
O
3i
O
O
O
O
O
O
IR spectra were measured on a Shimadzu IRAffinity-1 spec-
trophotometer. NMR spectra were recorded on a JEOL JNM-ECA
400 (400 MHz 1H, 100 MHz 13C), a JEOL JNM-ECA 500 (500 MHz
1H, 125 MHz 13C), JEOL JNM-ECA 700 (700 MHz 1H, 175 MHz
13C), or a JEOL JNM-ECA 800 (800 MHz 1H, 200 MHz 13C) spec-
trometer. Chemical shifts (d) and coupling constants (J) are given
in ppm and Hz, respectively. Low-resolution and high-resolution
mass spectra were recorded on a JEOL JMS-700T spectrometer.
Optical rotations were determined with a JASCO P-2200 polarime-
ter. Column chromatography was performed over Fuji Silysia silica
gel BW-200. All the organic extracts were dried over anhydrous
sodium sulfate prior to evaporation.
O
HO
HO
O
O
O
3j
3k
Scheme 1. Retrosynthetic analysis of homologs 1g–1k on acremomannolipin A
(1a).
the disc 1a. On the basis of the effect of 1a on the growth of
Dppb1
cells in the presence of MgCl2, the activities of homologs 1g–1k
were relatively evaluated by comparison of the size of colonies
around the discs (Fig. 4).
Upon addition of solutions of 1g and 1h, having two- and three-
carbon alditol chain, respectively, onto the filter paper disc (1g and
1h), smaller colonies formed around the discs than that around the
disc 1a which contained 1a, showing that the activities of 1g and
1h were significantly inferior to that of 1a, but remained to some
extents. On contrary, addition of a solution of the four-carbon side
3.1.1. 1-O-Acetyl-2,4-O-benzylidene-
D
-erythritol (7)
-erythritol7 (6, 1.00 g,
To a solution of 2,4-O-benzylidene-
D
4.76 mmol) in pyridine (9.5 mL) was added dropwise acetyl chlo-
ride (0.4 mL, 5.23 mmol) at 0 °C. After being stirred at 0 °C for
30 min, the reaction mixture was diluted with water. The resulting
mixture was extracted with EtOAc. The extract was washed with
brine and concentrated in vacuo. The residue was purified by col-
chain homolog 1i onto the filter paper (disc 1i),
Dppb1 cells more
easily grew in the presence of MgCl2, forming the bigger colonies
around disc than those around discs 1 g and 1 h. These results sug-
gested that a further increase of hydrophilicity of the alditol chain
might lead to increase of the cell proliferation. However, fair-sized
colonies were found around discs 1j and 1 k, which contained five-
and seven-carbon side chain homologs 1j and 1k, respectively. The
size of the colonies were nearly equal to that of disc 1a, indicating
that 1j and 1k were found as potent calcium signal modulators as
1a. Thus, derivatives bearing alditols of more than five carbons
were adequate for the calcium signal modulating. The length of
the alditol side chain was found to be a crucial determinant for
the activity.
umn chromatography (n-hexane–EtOAc, 10/1) to give 7 (1.06 g,
25
89%) as a colorless solid. [
a
]
D
29.4 (c 1.33, CHCl3). IR (neat)
cmꢀ1
:
3441, 1732, 1454, 1373, 1238, 1080, 1030. 1H NMR
(400 MHz, CDCl3) d: 2.15 (s, 3H, COCH3), 2.76 (d, J = 4.0 Hz, 1H,
OH), 3.64 (dd, J = 10.0, 9.6 Hz, 1H, H-1a), 3.68 (dddd, J = 10.0, 9.2,
4.0, 4.0 Hz, 1H, H-2), 3.77 (ddd, J = 9.2, 4.0, 2.4 Hz, 1H, H-3), 4.29
(dd, J = 12.4, 2.4 Hz, 1H, H-4a), 4.33 (dd, J = 9.6, 4.0 Hz, 1H, H-1b),
4.64 (dd, J = 12.4, 4.0 Hz, 1H, H-4b), 5.50 (s, 1H, OCHPh), 7.32–
7.40 (m, 3H, Ar), 7.47–7.50 (m, 2H, Ar). 13C NMR (100 MHz,
CDCl3) d: 20.9 (COCH3), 61.8 (C-2), 63.6 (C-4), 70.6 (C-1), 80.5
(C-3), 101.3 (OCHPh), 126.1 (2C)/128.3 (2C)/129.1 (d, Ar), 137.3
(s, Ar), 172.4 (COCH3). FABMS m/z (%): 253 ([M+H]+, 37), 55
OH
OH
OH
OH
OH
O
v
ii
iv
O
OR
RO
R1O
OH
OAc
OR2
O
O
OH
OBn
O
4 : R = H
3g : R = TBDMS
OH OH
Ph
i
9 : R1 = Bn, R2 = Ac
5
6 : R = H
7 : R = Ac
8
vi
vii
viii
10 : R1 = Bn, R2 = H
iii
11 : R1 = Bn, R2 = TBDMS
3i : R1 = H, R2 = TBDMS
O
OH
O
O
OH
O
OH
O
O
xiii
x
xi
O
RO
TBDPSO
OH
HO
O
RO
xii
HO
O
OH
OH
14
O
O
3k
12 : R = H
13 : R = TBDPS
15 : R = TBDPS
3j : R = H
ix
Scheme 2. Reagents and conditions: (i) Ref. 6; (ii) Ref. 7; (iii) AcCl, Py, 0 °C; (iv) Et3SiH, BF3ꢁEt2O, CH3CN, 0 °C; (v) DMP, PTSA, rt; (vi) NaOMe, MeOH, rt; (vii) TBDMSCl, NaH,
DMF, rt; (viii) H2, Pd-C, EtOAc, rt; (ix) TBDPSCl, imidazole, DMF, rt; (x) NaBH4, MeOH, rt; (xi) DMP, PTSA, acetone, rt; (xii) TBAF, THF, rt; (xiii) Ref. 9.

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N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
OR
O
5'
Ph
O
OCO(CH2)6CH3
O
iv
vi
RO
1'
O
O
BnO
1g
1h
1i
2'
RO
OTBDMS
3'
H
H
O
RO
H
OTBDMS
NOE
19g : R = H
16g : R = PMB
17g : R = H
v
ii
iii
20g : R = CO(CH2)4CH3
18g : R = CO(CH2)6CH3
OR
OCO(CH2)6CH3
O
Ph
O
O
RO
O
iv
O
O
vii
RO
O
O
BnO
O
O
RO
H
H
H
NOE
19h : R = H
16h: R = PMB
17h : R = H
v
ii
iii
20h : R = CO(CH2)4CH3
18h : R = CO(CH2)6CH3
OR
OCO(CH2)6CH3
O
RO
RO
Ph
O
O
O
O
O
iv
vii
i
O
BnO
O
RO
2
OTBDMS
H
OTBDMS
H
H
O
O
NOE
19i : R = H
16i : R = PMB
17i : R = H
v
ii
iii
20i : R = CO(CH2)4CH3
18i : R = CO(CH2)6CH3
OR
O
OCO(CH2)6CH3
O
Ph
O
O
O
O
RO
RO
RO
O
iv
iv
vii
O
O
O
BnO
O
1j
O
H
H
H
O
NOE
O
19j : R = H
16j : R = PMB
17j : R = H
v
ii
iii
20j : R = CO(CH2)4CH3
18j : R = CO(CH2)6CH3
OCO(CH2)6CH3
OR
O
RO
RO
RO
Ph
O
O
O
O
O
O
O
O
O
O
vii
O
O
BnO
O
1k
H
H
H
O
O
O
O
NOE
19k : R = H
20k : R = CO(CH2)4CH3
16k : R = PMB
17k : R = H
v
ii
iii
18k : R = CO(CH2)6CH3
Scheme 3. Reagents and conditions: (i) 3g–3k, Tf2O, DTBMP, CH2Cl2, ꢀ78 °C; (ii) DDQ, CH2Cl2, H2O, rt; (iii) CH3(CH2)6COCl, Py, DMAP, CH2Cl2, rt; (iv) H2, Pd-C, AcOEt, rt; (v)
[CH3(CH2)4CO]2O, Py, DMAP, CH2Cl2, rt; (vi) TBAF, THF, rt; (vii) 90% aq TFA aq, 0 °C.
(100). FABHRMS calcd for C13H17O5 [M+H]+: 253.1076; found:
253.1081.
Table 1
13C NMR data for acremomannolipin A (1a) and homologs 1g–1k in CD3OD
3.1.2. 1-O-Acetyl-4-O-benzyl-D-erythritol (8)
1aa
1gc
1hb
1ia
1jb
1ka
To a solution of 7 (784 mg, 3.11 mmol) in acetonitrile (155 mL)
was successively added triethylsilane (2.5 mL, 15.5 mmol) and
BF3ꢁEt2O (1.2 mL, 9.33 mmol) at 0 °C. The reaction mixture was
stirred at 0 °C for 30 min. After the reaction mixture was neutral-
ized with aqueous sodium hydrogen carbonate, the mixture was
extracted with EtOAc. The extract was washed with brine and con-
centrated in vacuo. The residue was purified by column chro-
C1
C2
C3
C4
C5
C6
C7
C10
C20
C30
C40
C50
C60
73.7
71.7
71.0
71.2
73.0
65.2
73.3
61.9
72.5
72.2
64.3
73.1
72.4
73.6
64.6
73.50
71.69
72.1
71.67
64.8
72.8
69.6
71.1
70.3
71.3
73.3
65.2
100.0
70.5
72.7
66.8
73.5
63.0
100.5
70.5
72.7
66.8
73.5
63.0
99.4
68.6
70.7
65.7
72.6
62.4
100.3
70.4
72.7
66.9
73.5
63.1
100.3
70.5
72.7
66.8
73.5
63.1
100.3
70.5
72.7
66.8
73.48
63.1
matography (n-hexane–EtOAc, 5/1) to give 8 (184 mg, 23%) as a
24
colorless oil. [
a
]
D
ꢀ5.7 (c 0.62, CHCl3). IR (neat) cmꢀ1: 3422,
1718, 1452, 1368, 1246, 1074, 1043, 1028. 1H NMR (400 MHz,
CD3OD) d: 1.96 (s, 3H, COCH3), 3.48 (dd, J = 11.2, 5.6 Hz, 1H, H-
1a), 3.59 (dd, J = 11.2, 3.2 Hz, 1H, H-1b), 3.62 (ddd, J = 7.2, 5.6,
3.2 Hz, 1H, H-2), 3.69 (ddd, J = 7.2, 6.4, 3.2 Hz, 1H, H-2), 4.02 (dd,
J = 11.6, 6.4 Hz, 1H, H-4a), 4.19 (dd, J = 11.6, 3.2 Hz, 1H, H-4b),
4.47 (s, 2H, OCH2Ph), 7.14–7.28 (m, 5H, Ar). 13C NMR (100 MHz,
a
b
c
Measured at 200 MHz.
Measured at 175 MHz.
Measured at 125 MHz.

N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
3765
25
(154 mg, 98%) as a colorless oil. [
a]
D
ꢀ9.8 (c 0.20, CHCl3). IR (neat)
cmꢀ1: 3456, 1454, 1371, 1248, 1217, 1167, 1076, 1053. 1H NMR
(400 MHz, CDCl3) d: 1.36/1.43 [each s, 3H, C(CH3)2], 2.59 (dd,
J = 6.4, 6.4 Hz, 1H, OH), 3.57 (dd, J = 9.6, 4.8 Hz, 1H, H-4a), 3.65
(dd, J = 9.6, 7.6 Hz, 1H, H-4b), 3.70 (ddd, J = 12.0, 6.4, 6.0 Hz, 1H,
H-1a), 3.74 (ddd, J = 12.0, 6.4, 6.0 Hz, 1H, H-1b), 4.31 (ddd, J = 6.0,
6.0, 6.0 Hz, 1H, H-2), 4.39 (ddd, J = 7.6, 6.0, 4.8 Hz, 1H, H-3), 4.55/
4.59 (each d, J = 12.0 Hz, 1H, OCH2Ph), 7.29–7.39 (m, 5H, Ar). 13C
NMR (100 MHz, CDCl3) d: 25.1/27.6 [C(CH3)2], 60.8 (C-1), 68.2
(C-4), 73.7 (OCH2Ph), 75.3 (C-3), 77.1 (C-2), 108.4 [C(CH3)2],
127.7 (2C)/127.9/128.5 (2C) (d, Ar), 137.1 (s, Ar). FABMS m/z (%):
253 ([M+H]+, 4), 91 (100). FABHRMS calcd for C14H21O4 [M+H]+:
253.1440; found: 253.1447.
3.1.5. 4-O-Benzyl-1-O-(tert-butyldimethylsilyl)-2,3-O-
isopropylidene-
D
-erythritol (11)
(150 mg, 0.595 mmol) and imidazole
To solution of
a
7
(80.9 mg, 1.19 mmol) in DMF (6.0 mL) was added TBSCl (179 mg,
1.19 mmol) at 0 °C. The reaction mixture was stirred at room tem-
perature for 4 h. After being cooled with ice-water, the reaction
mixture was diluted with water. The resulting mixture was
extracted with EtOAc. The extract was washed with brine and con-
centrated in vacuo. The residue was purified by column chro-
Figure 4. Suppression of the Clꢀ-sensitive growth defect of calcineurin deletion
cells by acremomannolipin A (1a) and the synthesized homologs 1g–1k. Calcineurin
matography (n-hexane–EtOAc, 20/1) to give 8 (210 mg, 96%) as a
24
colorless oil. [
a]
ꢀ3.3 (c 0.92, CHCl3). IR (neat) cmꢀ1: 1454,
deletion (
Filter paper containing DMSO (5
1j and 1k contain 5 l of 100
D
ppb1) cells were spread onto the YPD medium containing 0.11 M MgCl2.
l) is used as a control. Filter papers 1a, 1g, 1h, 1i,
M DMSO solution of 1a, 1g, 1h, 1i, 1j and 1k,
D
1379, 1371, 1252, 1215, 1111, 1094, 1055. 1H NMR (400 MHz,
CDCl3) d: 0.01/0.02 [each s, 3H, Si(CH3)2], 0.84 [s, 9H, SiC(CH3)3],
1.36/1.44 [each s, 3H, C(CH3)2], 3.56 (dd, J = 10.4, 7.6 Hz, 1H, H-
4a), 3.58 (dd, J = 10.4, 4.8 Hz, 1H, H-1a), 3.65 (dd, J = 10.4, 7.2 Hz,
1H, H-1b), 3.72 (dd, J = 10.4, 3.6 Hz, 1H, H-4b), 4.13 (ddd, J = 7.2,
6.4, 4.8 Hz, 1H, H-2), 4.39 (ddd, J = 7.6, 6.4, 3.6 Hz, 1H, H-3), 4.52/
4.66 (each d, J = 12.0 Hz, 1H, CH2Ph), 7.26–7.34 (m, 5H, Ar). 13C
NMR (100 MHz, CDCl3) d: ꢀ5.4/ꢀ5.5 [Si(CH3)2], 18.1 [SiC(CH3)3],
25.3/27.9 [C(CH3)2], 25.8 [3C, SiC(CH3)3], 61.7 (C-1), 68.7 (C-4),
73.5 (OCH2Ph), 76.5 (C-2), 77.0 (C-3), 108.6 [C(CH3)2], 127.6/
127.8 (2C)/128.4 (2C) (d, Ar), 138.0 (s, Ar). FABMS m/z (%): 367
([M+Na]+, 14), 91 (100). FABHRMS calcd for C20H35O4Si [M+H]+:
367.2305; found: 367.2301.
l
l
l
respectively. Cells were grown at 27 °C for 3 days.
CD3OD) d: 20.8 (COCH3), 67.2 (C-4), 71.1/72.1 (C-2 and C-3), 72.7
(C-1), 74.4 (OCH2Ph), 128.6/128.9 (2C)/129.3 (2C) (d, Ar), 139.7
(s, Ar), 173.0 (COCH3). FABMS m/z (%): 277 ([M+Na]+, 5), 55
(100). FABHRMS calcd for C13H18O5Na [M+H]+: 277.1052; found:
277.1059.
3.1.3. 1-O-Acetyl-4-O-benzyl-2,3-O-isopropylidene-D-erythritol
(9)
To a mixture of 8 (183 mg, 0.718 mmol) and 2,2-dimethoxypro-
pane (DMP, 5.0 mL) was added p-toluenesulfonic acid (13.7 mg,
0.0718 mmol) at room temperature, and the reaction mixture
was stirred at room temperature for 1 h. The reaction was
quenched with Et3N, and the resulting mixture was concentrated
in vacuo. The residue (245 mg) was purified by column chro-
3.1.6. 1-O-(tert-Butyldimethylsilyl)-2,3-O-isopropylidene-D-
erythritol (3i)
A suspension of 11 (177 mg, 0.482 mmol) and 10% palladium on
carbon (30 mg) in EtOAc (4.8 mL) was hydrogenated at room tem-
perature until the uptake of hydrogen ceased. The catalyst was fil-
tered off and washed with EtOAc. The combined filtrate and
washings were concentrated in vacuo. The residue was purified
by column chromatography (n-hexane–EtOAc, 10/1) to give 3i
matography (n-hexane–EtOAc, 15/1) to give 9 (203 mg, 96%) as a
24
colorless oil. [
a]
ꢀ29.4 (c 0.73, CHCl3). IR (neat) cmꢀ1: 1744,
D
1454, 1371, 1236, 1219, 1069, 1090, 1043. 1H NMR (400 MHz,
CDCl3) d: 1.38/1.47 [each s, 3H, C(CH3)2], 2.07 (s, 3H, COCH3),
3.51–3.57 (m, 2H, H-4), 4.05 (dd, J = 11.6, 7.2 Hz, 1H, H-1a), 4.31
(dd, J = 11.6, 3.6 Hz, 1H, H-1b), 4.37 (ddd, J = 7.2, 6.4, 3.6 Hz, 1H,
H-2), 4.39 (ddd, J = 6.4, 6.4, 6.4 Hz, 1H, H-3), 4.53/4.59 (each d,
J = 12.0 Hz, 1H, OCH2Ph), 7.28–7.37 (m, 5H, Ar). 13C NMR
(100 MHz, CDCl3) d: 20.7 (COCH3), 25.2/27.6 [C(CH3)2], 63.0 (C-1),
68.0 (C-4), 73.5 (OCH2Ph), 74.7/75.3 (C-2 and C-3), 109.0
[C(CH3)2], 127.69/127.74 (2C)/128.3 (2C) (d, Ar), 137.5 (s, Ar),
170.6 (COCH3). FABMS m/z (%): 295 ([M+H]+, 3), 91 (100).
FABHRMS calcd for C16H23O5 [M+H]+: 295.1545; found: 295.1526.
(123 mg, 92%) as a colorless oil. [a]
23 +7.8 (c 1.16, CHCl3). IR (neat)
D
cmꢀ1: 3483, 1470, 1373, 1254, 1219, 1169, 1084, 1057. 1H NMR
(500 MHz, CDCl3) d: 0.107/0.111 [each s, 3H, Si(CH3)2], 0.90 [s,
9H, SiC(CH3)3], 1.36/1.42 [each s, 3H, C(CH3)2], 3.01 (dd, J = 8.3,
5.4 Hz, 1H, OH), 3.68 (dd, J = 10.6, 4.0 Hz, 1H, H-1a), 3.76 (ddd,
J = 11.8, 8.3, 5.5 Hz, 1H, H-4a), 3.79 (dd, J = 10.6, 9.2 Hz, 1H, H-
1b), 3.82 (ddd, J = 11.8, 7.5, 5.4 Hz, 1H, H-4b), 4.23 (ddd, J = 9.2,
6.0, 4.0 Hz, 1H, H-2), 4.35 (ddd, J = 7.5, 6.0, 5.5 Hz, 1H, H-3). 13C
NMR (125 MHz, CDCl3) d: ꢀ5.6/ꢀ5.5 [Si(CH3)2], 18.2 [SiC(CH3)3],
25.2/27.8 [C(CH3)2], 25.8 [3C, SiC(CH3)3], 60.8 (C-4), 61.5 (C-1),
76.8 (C-2), 77.3 (C-3), 108.4 [C(CH3)2]. FABMS m/z (%): 277
([M+H]+, 26), 73 (100). FABHRMS calcd for C13H29O4Si [M+H]+:
277.1835; found: 277.1831.
3.1.4. 4-O-Benzyl-2,3-O-isopropylidene-D-erythritol (10)
To a solution of 9 (184 mg, 0.626 mmol) in MeOH (6.3 mL) was
added sodium methoxide (33.8 mg, 0.626 mmol) at room tempera-
ture. After being stirred at room temperature for 30 min, the reac-
tion mixture was neutralized with Amberlite IR-120. The resin was
filtered off and washed with MeOH. The combined filtrate and
washings were concentrated in vacuo. The residue was purified
by column chromatography (n-hexane–EtOAc, 10/1) to give 10
3.1.7. 5-O-(tert-Butyldiphenylsilyl)-1,2:3,4-di-O-isopropylidene-
D-arabinitol (15)
To a solution of D-arabinose (12) (300 mg, 2.00 mmol) and imi-
dazole (272 mg, 4.00 mmol) in DMF (15 mL) was added dropwise a
solution of tert-butyldiphenylsilyl chloride (TBDPSCl, 0.57 mL,

3766
N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
2.20 mmol) in DMF (5 mL) at 0 °C. The reaction mixture was stirred
at room temperature for 23 h. After being cooled with ice-water,
the reaction mixture was diluted with water. The resulting mixture
was extracted with EtOAc. The extract was washed with brine, and
concentrated to give a pale yellow oil (13, 828 mg). To a mixture of
13 (828 mg) and MeOH (15 mL) was added NaBH4 (113 mg,
3.00 mmol), and the reaction mixture was stirred at room tem-
perature for 30 min. After the reaction was quenched with acetone,
the solvent was evaporated, and the residue 14 was dissolved in
DMP (15 mL). To the solution was added p-toluenesulfonic acid
(500 mg), and the reaction mixture was stirred at room tempera-
ture for 2 h. The reaction was quenched with Et3N, and the result-
ing mixture was concentrated in vacuo. The residue was purified
IR (neat) cmꢀ1:1512, 1454, 1385, 1250, 1096. 1H NMR (800 MHz,
CDCl3) d: 0.06/0.07 [each s, 3H, Si(CH3)2], 0.89 [s, 9H, SiC(CH3)3],
3.31 (ddd, J = 10.3, 9.8, 4.8 Hz, 1H, H-50), 3.55 (dd, J = 9.8, 3.2 Hz,
1H, H-30), 3.63 (ddd, J = 10.5, 7.8, 4.6 Hz, 1H, H-1a), 3.77 (ddd,
J = 11.0, 4.6, 4.1 Hz, 1H, H-2a), 3.80 (s, 3H, OCH3), 3.82 (ddd,
J = 11.0, 7.8, 4.4 Hz, 1H, H-2b), 3.926 (dd, J = 10.3, 10.3 Hz, 1H, H-
60a), 3.933 (ddd, J = 10.5, 4.4, 4.1 Hz, 1H, H-1b), 3.94 (dd, J = 3.2,
0.7 Hz, 1H, H-20), 4.18 (dd, J = 9.8, 9.8 Hz, 1H, H-40), 4.30 (dd,
J = 10.3, 4.8 Hz, 1H, H-60b), 4.54/4.64 (each d, J = 12.4 Hz, 1H,
OCH2Ph), 4.56 (d, J = 0.7 Hz, 1H, H-10), 4.81/4.90 (each d,
J = 11.6 Hz, 1H, OCH2C6H4OCH3), 5.61 (s, 1H, CHPh), 6.84 (d,
J = 8.8 Hz, 2H, Ar), 7.25ꢀ7.30 (m, 5H, Ar), 7.34ꢀ7.38 (m, 3H, Ar),
7.38 (d, J = 8.8 Hz, 2H, Ar), 7.49ꢀ7.50 (m, 2H, Ar). 13C NMR
(200 MHz, CDCl3) d: ꢀ5.32/ꢀ5.30 [Si(CH3)2], 18.3 [SiC(CH3)3],
25.9 [3C, SiC(CH3)3], 55.2 (OCH3), 62.7 (C-2), 67.6 (C-50), 68.6 (C-
60), 71.1 (C-1), 72.2 (OCH2Ph), 74.3 (OCH2C6H4OCH3), 75.3 (C-20),
77.8 (C-30), 78.6 (C-40), 101.4 (CHPh), 102.7 (C-10), 113.5 (2C)/
126.0 (2C)/127.5 (3C)/128.2 (2C)/128.3 (2C)/128.8/130.3 (2C) (d,
Ar), 130.6/137.6/138.3/159.2 (s, Ar). FAB MS m/z (%): 659
([M+Na]+, 7), 121 (100). FAB HRMS calcd for C36H48O8SiNa
[M+Na]+: 659.3016; found: 659.3041.
by column chromatography (n-hexane–EtOAc, 30/1) to give 15
25
(379 mg, 40% from 12) as a colorless oil. [
a
]
D
+4.80 (c 1.27,
CHCl3). IR (neat) cmꢀ1
:
1250, 1215, 1111, 1072. 1H NMR
(400 MHz, CDCl3) d: 1.06 [s, 9H, C(CH3)3], 1.35/1.36.1.42.1.46 [each
s, 3H, C(CH3)2], 3.62 (dd, J = 10.8, 4.0 Hz, 1H, H-5a), 3.73 (dd, J = 8.4,
6.4 Hz, 1H, H-1a), 3.81 (dd, J = 10.8, 7.6 Hz, 1H, H-5b), 4.08 (dd,
J = 8.4, 6.8 Hz, 1H, H-1b), 4.16 (dd, J = 6.4, 6.0 Hz, 1H, H-3), 4.19
(ddd, J = 7.6, 6.4, 4.0 Hz, 1H, H-4), 4.41 (ddd, J = 6.8, 6.4, 6.0 Hz,
1H, H-2), 7.37–7.47 (m, 6H, Ar), 7.63–7.67 (m, 4H, Ar). 13C NMR
(100 MHz, CDCl3) d: 19.2 [SiC(CH3)3], 25.2/25.4/26.8/27.6
[C(CH3)2], 26.9 [3C, SiC(CH3)3], 62.7 (C-5), 66.7 (C-1), 74.3 (C-2),
76.9 (C-4), 78.5 (C-3), 109.0/109.6 [C(CH3)2], 127.79 (2C)/129.81
(2C)/129.86/129.90/135.5 (4C) (d, Ar), 132.8/132.9 (s, Ar). FABMS
m/z (%): 493 ([M+Na]+, 13), 135 (100). FABHRMS calcd for
Following the method described above, (R)-(ꢀ)-2,2-dimethyl-
1,3-dioxolane-4-methanol (3h, 476 mg, 3.60 mmol), 3i (116 mg,
0.421 mmol), 3j (476 mg, 2.05 mmol), and 3k (300 mg, 0.903 mmol)
were converted to the corresponding coupled products, 1,
2-O-isopropylidene-
D-glycerol-3-yl
3-O-benzyl-4,6-O-benzyli-
dene-2-O-p-methoxybenzyl-b-
D-mannopyranoside (16h, 692 mg,
C
27H38O5SiNa [M+Na]+: 493.2386; found: 493.2387.
40%), 2,3:4,5-di-O-isopropylidene-
dimethylsilyl)-2,3-O-isopropylidene-
6-O-benzylidene-2-O-p-methoxybenzyl-b-
161 mg, 62%), 1,2:3,4-di-O-isopropylidene-
zyl-4,6-O-benzylidene-2-O-p-methoxybenzyl-b-
(16j, 620 mg, 52%), and 2,3:4,5:6,7-tri-O-isopropylidene-
galactoheptitol-1-yl 3-O-benzyl-4,6-O-benzylidene-2-O-p-methoxy-
D
-arabinitol-1-yl 1-O-(tert-butyl-
-erythritol-4-yl 3-O-benzyl-4,
-mannopyranoside (16i,
-arabinitol-5-yl 3-O-ben-
-mannopyranoside
-glycero-
D
3.1.8. 1,2:3,4-Di-O-isopropylidene- -arabinitol (3j)
D
D
To a solution of 15 (379 mg, 0.805 mmol) in THF (6.9 mL) was
added 1 M solution of tetrabutylammonium fluoride (TBAF) in
THF (1.2 mL, 1.21 mmol) at 0 °C. After being stirred at room tem-
perature for 5 h, the reaction mixture was diluted with water.
The resulting mixture was extracted with EtOAc. The extract was
washed with brine, and concentrated in vacuo. The residue was
D
D
D
D-
benzyl-b-
D-mannopyranoside (16k, 250 mg, 42%), respectively.
24
16h: A colorless oil. [
a]
ꢀ62.3 (c 0.79, CHCl3). IR (neat) cmꢀ1
:
D
purified by column chromatography (n-hexane–EtOAc, 4/1) to give
1612, 1512, 1454, 1373, 1250, 1215, 1096, 1053. 1H NMR
(700 MHz, CDCl3) d: 1.38/1.42 [each s, 3H, C(CH3)2], 3.32 (ddd,
J = 10.2, 9.6, 4.8 Hz, 1H, H-50), 3.57 (dd, J = 9.6, 3.2 Hz, 1H, H-30),
3.59 (dd, J = 10.0, 6.2 Hz, 1H, H-3a), 3.75 (dd, J = 8.4, 6.2 Hz, 1H,
H-1a), 3.80 (s, 3H, OCH3), 3.88 (dd, J = 10.0, 5.8 Hz, 1H, H-3b),
3.93 (dd, J = 10.2, 10.2 Hz, 1H, H-60a), 3.96 (dd, J = 3.2, 0.8 Hz, 1H,
H-20), 4.09 (dd, J = 8.4, 6.4 Hz, 1H, H-1b), 4.19 (dd, J = 9.6, 9.6 Hz,
1H, H-40), 4.29 (dddd, J = 6.4, 6.2, 6.2, 5.8 Hz, 1H, H-2), 4.30 (dd,
J = 10.2, 4.8 Hz, 1H, H-60b), 4.52 (d, J = 0.8 Hz, 1H, H-10), 4.58/4.68
(each d, 1H, J = 12.4 Hz, OCH2Ph), 4.81/4.88 (each d, 1H,
J = 11.8 Hz, OCH2C6H4OCH3), 5.61 (s, 1H, CHPh), 6.85 (d,
J = 8.6 Hz, 2H, Ar), 7.26–7.31 (m, 5H, Ar), 7.34–7.39 (m, 5H, Ar),
7.49–7.50 (m, 2H, Ar). 13C NMR (175 MHz, CDCl3) d: 25.4/26.8
[C(CH3)2], 55.3 (OCH3), 67.0 (C-1), 67.7 (C-50), 68.5 (C-60), 70.9 (C-
3), 72.4 (OCH2Ph), 74.3 (OCH2C6H4OCH3), 74.7 (C-2), 75.1 (C-20),
77.8 (C-30), 78.6 (C-40), 101.4 (CHPh), 102.2 (C-10), 109.6
[C(CH3)2], 113.5 (2C)/126.0 (2C)/127.52 (2C)/127.55/128.2 (2C)/
128.3 (2C)/128.8/130.2 (2C) (d, Ar), 130.5/137.5/138.3/159.2 (s,
Ar). FABMS m/z (%): 615 ([M+Na]+, 13), 57 (100). FABHRMS calcd
25
3j (128 mg, 68%) as a colorless amorphous. [
a
]
+7.03 (c 0.69,
D
CHCl3). IR (neat) cmꢀ1: 3406, 1373, 1250, 1215, 1161, 1049. 1H
NMR (700 MHz, CDCl3) d: 1.388/1.391/1.47/1.52 [each s, 3H,
C(CH3)2], 2.33 (dd, J = 7.2, 5.2 Hz, 1H, OH), 3.66 (ddd, J = 11.8, 5.2,
5.2 Hz, 1H, H-5a), 3.70 (ddd, J = 11.8, 7.2, 5.2 Hz, 1H, H-5b), 3.81
(dd, J = 8.2, 7.0 Hz, 1H, H-1a), 4.13 (dd, J = 8.2, 6.6 Hz, 1H, H-1b),
4.15 (dd, J = 6.4, 5.6 Hz, 1H, H-3), 4.20 (ddd, J = 6.4, 5.2, 5.2 Hz,
1H, H-4), 4.32 (ddd, J = 7.0, 6.6, 5.6 Hz, 1H, H-2). 13C NMR
(175 MHz, CDCl3) d: 25.3/25.4/26.3/27.4 [C(CH3)2], 61.4 (C-5),
66.4 (C-1), 74.0 (C-2), 77.0 (C-4), 77.5 (C-3), 109.1/110.0
[C(CH3)2]. FAB MS m/z (%): 233 ([M+H]+, 35), 59 (100). FAB HRMS
calcd for C11H21O5 [M+H]+: 233.1389; found: 233.1402.
3.1.9. Coupling reaction between mannosyl sulfoxide (2) and
protected alditols (3g–3k)
Under argon atmosphere, to
0.852 mmol) and 2,6-di-tert-butyl-4-methylpyridine (DTBMP,
350 mg, 1.70 mmol) in CH2Cl2 (8.5 mL) was added
a
solution of 24b (500 mg,
trifluoromethanesulfonic anhydride (Tf2O, 0.15 mL, 0.938 mmol)
at ꢀ78 °C, and the mixture was stirred at ꢀ78 °C for 10 min. A
solution of 3g6 (165 mg, 0.938 mmol) in CH2Cl2 (8.5 mL) was added
to the mixture, and the resulting mixture was stirred at ꢀ78 °C for
another 1 h. After addition of aqueous sodium hydrogen carbonate,
the resulting mixture was extracted with CH2Cl2. The extract was
washed with brine, and condensed in vacuo. The residue was
purified by column chromatography (n-hexane–EtOAc, 12/1) to
give 2-(tert-butyldimethylsiloxy)ethanol-1-yl 3-O-benzyl-4,6-
for C34H40O9Na [M+Na]+: 615.2570; found: 615.2593.
25
16i: A colorless oil. [
a
]
D
ꢀ34.1 (c 0.73, CHCl3). IR (neat) cmꢀ1
:
1612, 1512, 1454, 1373, 1304, 1250, 1219, 1173, 1111, 1057. 1H
NMR (700 MHz, CDCl3) d: 0.039/0.042 [each s, 3H, Si(CH3)2], 0.87
[s, 9H, SiC(CH3)3], 1.38/1.45 [each s, 3H, C(CH3)2], 3.31 (ddd,
J = 10.0, 9.6, 4.8 Hz, 1H, H-50), 3.565 (dd, J = 9.6, 3.2 Hz, 1H, H-30),
3.573 (dd, J = 10.5, 4.8 Hz, 1H, H-1a), 3.60 (dd, 11.0, 8.6 Hz, 1H,
H-4a), 3.62 (dd, J = 10.5, 7.6 Hz, 1H, H-1b), 3.80 (s, 3H, OCH3),
3.92 (dd, J = 10.4, 10.0 Hz, 1H, H-60a), 4.01 (d, J = 3.2 Hz, 1H, H-20),
4.13 (ddd, J = 7.6, 6.2, 4.8 Hz, 1H, H-2), 4.19 (dd, J = 9.6, 9.6 Hz,
1H, H-40), 4.27 (dd, J = 10.4, 4.8 Hz, 1H, H-60b), 4.29 (dd, J = 11.0,
O-benzylidene-2-O-p-methoxybenzyl-b-
D
25
-mannopyranoside (16g,
ꢀ47.4 (c 1.08, CHCl3).
397 mg, 73%) as a colorless oil. [a]
D

N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
3767
2.4 Hz, 1H, H-4b), 4.44 (ddd, J = 8.6, 6.2, 2.4 Hz, 1H, H-3), 4.55 (s,
1H, H-10), 4.58/4.66 (each d, J = 12.4 Hz, 1H, OCH2Ph), 4.85/4.91
(each d, J = 11.8 Hz, 1H, OCH2C6H4OCH3), 5.62 (s, 1H, CHPh), 6.85
(d, J = 8.6 Hz, 2H, Ar), 7.25–7.33 (m, 5H, Ar), 7.33–7.39 (m, 3H,
Ar), 7.41 (d, J = 8.6 Hz, 2H, Ar), 7.49–7.50 (m, 2H, Ar). 13C NMR
(175 MHz, CDCl3) d: ꢀ5.6/ꢀ5.5 [Si(CH3)2], 18.1 [SiC(CH3)3], 25.4/
28.0 [C(CH3)2], 25.8 [3C, SiC(CH3)3], 55.2 (OCH3), 61.6 (C-1), 67.6
(C-50), 68.56/68.58 (C-4 and C-60), 72.2 (OCH2Ph), 74.3
(OCH2C6H4OCH3), 74.9 (C-20), 76.7 (C-2), 77.0 (C-3), 77.7 (C-30)
78.6 (C-40), 101.4 (CHPh), 102.2 (C-10), 108.6 [C(CH3)2], 113.5
(2C)/126.0 (2C)/127.51 (2C)/127.53/128.2 (2C)/128.3 (2C)/128.8/
130.3 (2C) (d, Ar), 130.5/137.6/138.3/159.2 (s, Ar). FABMS m/z
(%): 759 ([M+Na]+, 6), 121 (100). FABHRMS calcd for
(DDQ, 191 mg, 0.808 mmol), and the mixture was stirred at room
temperature for 1 h. After addition of aqueous sodium hydrogen
carbonate, the resulting mixture was extracted with CH2Cl2. The
extract was washed with brine, and concentrated in vacuo. The
residue was purified by column chromatography (n-hexane–
EtOAc, 1/1) to give 2-(tert-butyldimethylsiloxy)ethanol-1-yl 3-O-
benzyl-4,6-O-benzylidene-b-D-mannopyranoside (17k, 214 mmol,
25
72%) as a colorless oil. [
a]
ꢀ18.0 (c 0.99, CHCl3). IR (neat) cmꢀ1
:
D
3451, 1385, 1256, 1094. 1H NMR (400 MHz, CDCl3) d: 0.05/0.06
[each s, 3H, Si(CH3)2], 0.88 [s, 9H, SiC(CH3)3], 3.34 (ddd, J = 9.6,
9.6, 4.8 Hz, 1H, H-50), 3.63 (dd, J = 9.6, 3.2 Hz, 1H, H-30), 3.71
(ddd, J = 10.4, 7.6, 4.0 Hz, 1H, H-1a), 3.76 (ddd, J = 11.2, 4.0,
4.0 Hz, 1H, H-2a), 3.83 (ddd, J = 11.2, 7.6, 4.4 Hz, 1H, H-2b), 3.89
(dd, J = 10.4, 9.6 Hz, 1H, H-60a), 3.91 (ddd, J = 10.4, 4.4, 4.0 Hz, 1H,
H-1b), 4.14 (dd, J = 3.2, 0.8 Hz, 1H, H-20), 4.15 (dd, J = 9.6, 9.6 Hz,
1H, H-40), 4.33 (dd, J = 10.4, 4.8 Hz, 1H, H-60b), 4.63 (d, J = 0.8 Hz,
1H, H-10), 4.78/4.86 (each d, J = 12.0 Hz, 1H, OCH2Ph), 5.61 (s, 1H,
CHPh), 7.27ꢀ7.41 (m, 8H, Ar), 7.49ꢀ7.52 (m, 2H, Ar). 13C NMR
(100 MHz, CDCl3) d: ꢀ5.4/ꢀ5.3 [Si(CH3)2], 18.2 [SiC(CH3)3], 25.8
[3C, SiC(CH3)3], 62.6 (C-2), 66.9 (C-50), 68.6 (C-60), 70.0 (C-20),
70.8 (C-1), 72.5 (OCH2Ph), 76.7 (C-30), 78.4 (C-40), 100.7, (C-10)
101.5 (CHPh), 126.0 (2C)/127.8/127.9 (2C)/128.2 (2C)/128.4 (2C)/
128.9 (d, Ar), 137.4/137.9 (s, Ar). FABMS m/z (%): 539 ([M+Na]+,
C41H56O10SiNa [M+Na]+: 759.3540; found: 759.3548.
23
16j: A colorless oil. [
a]
ꢀ43.6 (c 0.19, CHCl3). IR (neat) cmꢀ1
:
D
1377, 1250, 1219, 1092, 1049. 1H NMR (700 MHz, CDCl3) d: 1.34/
1.40/1.46/1.49 [each s, 3H, C(CH3)2], 3.32 (ddd, J = 9.8, 9.8, 4.8 Hz,
1H, H-50), 3.58 (dd, J = 9.8, 3.0 Hz, 1H, H-30), 3.60 (dd, J = 10.0,
6.2 Hz, 1H, H-5a), 3.70 (dd, J = 8.4, 7.1 Hz, 1H, H-1a), 3.80 (s, 3H,
OCH3), 3.89 (dd, J = 10.2, 9.8 Hz, 1H, H-60a), 3.95 (dd, J = 3.0,
0.8 Hz, 1H, H-20), 3.98 (dd, J = 10.0, 6.4 Hz, 1H, H-5b), 4.126 (dd,
J = 6.4, 6.4 Hz, 1H, H-3), 4.132 (dd, J = 8.4, 6.6 Hz, 1H, H-1b), 4.18
(dd, J = 9.8, 9.8 Hz, 1H, H-40), 4.22 (ddd, J = 7.1, 6.6, 6.4 Hz, 1H, H-
2), 4.296 (ddd, J = 6.4, 6.4, 6.2 Hz, 1H, H-4), 4.300 (dd, J = 10.2,
4.8 Hz, 1H, H-60b), 4.50 (d, J = 0.8 Hz, 1H, H-10), 4.58/4.70 (each d,
J = 12.4 Hz, 1H, OCH2Ph), 4.82 (s, 2H, OCH2C6H4OCH3), 5.62 (s,
1H, CHPh), 6.85 (d, J = 8.8 Hz, 2H, Ar), 7.26–7.32 (m, 5H, Ar),
7.34–7.39 (m, 5H, Ar), 7.49–7.50 (m, 2H, Ar). 13C NMR (175 MHz,
CDCl3) d: 25.3/25.5/26.6/27.6 [C(CH3)2], 55.2 (OCH3), 66.3 (C-1),
67.6 (C-50), 68.1 (C-5), 68.6 (C-60), 72.5 (OCH2Ph), 74.2 (C-2), 74.5
(OCH2C6H4OCH3), 75.5 (2C, C-4 and C-20), 77.7 (C-3), 77.9 (C-30),
78.6 (C-40), 101.4 (CHPh), 101.9 (C-10), 109.2/109.6 [C(CH3)2],
113.5 (2C)/126.0 (2C)/127.5 (2C)/127.6/128.2 (2C)/128.3 (2C)/
128.9/130.2 (2C) (d, Ar), 130.4/137.5/138.3/159.2 (s, Ar). FABMS
m/z (%): 715 ([M+Na]+, 7), 121 (100). FABHRMS calcd for
50), 91 (100). FABHRMS calcd for
539.2441; found: 539.2432.
C
28H40O7SiNa [M+Na]+:
Following the method described above, 16h (680 mg, 1.15 mmol),
16i (134 mg, 0.182 mmol), 16j (596 mg, 0.860 mmol), and 16k
(229 mg, 0.289 mmol) were converted to 1,2-O-isopropylidene-
glycerol-3-yl 3-O-benzyl-4,6-O-benzylidene-b- -mannopyranoside
(17h, 490 mg, 90%), 1-O-(tert-butyldimethylsilyl)-2,3-O-isopro-
pylidene- -erythritol-4-yl 3-O-benzyl-4,6-O-benzylidene-b- -man-
nopyranoside (17i, 91.0 mg, 81%), 1,2:3,4-di-O-isopropylidene- -ara-
binitol-5-yl 3-O-benzyl-4,6-O-benzylidene-b- -mannopyranoside
(17j, 453 mg, 92%), and 2,3:4,5:6,7-tri-O-isopropylidene- -glycero-
-galactoheptitol-1-yl 3-O-benzyl-4,6-O-benzylidene-b- -mannopyr-
D-
D
D
D
D
D
D
D
D
C
39H48O11Na [M+Na]+: 715.3094; found: 715.3069.
anoside (17k, 165 mg, 85%), respectively.
24
25
16k: A colorless amorphous. [
a]
ꢀ44.3 (c 1.06, CHCl3). IR (neat)
D
17h: A colorless oil. [
a]
ꢀ20.7 (c 0.51, CHCl3). IR (neat) cmꢀ1
:
D
cmꢀ1
:
1512, 1454, 1377, 1250, 1215, 1092, 1049. 1H NMR
3483, 1454, 1373, 1258, 1211, 1153, 1096, 1049. 1H NMR
(400 MHz, CDCl3) d: 1.36/1.42 [each s, 3H, C(CH3)2], 2.04 (br s,
1H, OH), 3.35 (ddd, J = 10.0, 9.6, 4.8 Hz, 1H, H-50), 3.65 (dd,
J = 9.6, 3.2 Hz, 1H, H-30), 3.66 (dd, J = 10.8, 6.8 Hz, 1H, H-3a), 3.70
(dd, J = 8.4, 6.0 Hz, 1H, H-1a), 3.88 (dd, J = 10.8, 4.8 Hz, 1H, H-3b),
3.89 (dd, J = 10.4, 10.0 Hz, 1H, H-60a), 4.08 (dd, J = 8.4, 6.4 Hz, 1H,
H-1b), 4.14 (dd, J = 9.6, 9.6 Hz, 1H, H-40), 4.19 (d, J = 3.2 Hz, 1H,
H-20), 4.33 (dd, J = 10.4, 4.8 Hz, 1H, H-60b), 4.34 (dddd, J = 6.8, 6.4,
6.0, 4.8 Hz, 1H, H-2), 4.60 (s, 1H, H-10), 4.78/4.86 (each d,
J = 12.4 Hz, 1H, OCH2Ph), 5.61 (s, 1H, CHPh), 7.27–7.42 (m, 8H,
Ar), 7.49–7.51 (m, 2H, Ar). 13C NMR (100 MHz, CDCl3) d: 25.3/
26.8 [C(CH3)2], 66.7 (C-1), 67.0 (C-50), 68.5 (C-60), 69.7 (C-20), 70.9
(C-3), 72.5 (OCH2Ph), 74.7 (C-2), 76.7 (C-30), 78.3 (C-40), 100.4 (C-
10), 101.6 (CHPh), 109.8 [C(CH3)2], 126.0 (2C)/127.9 (3C)/ 128.2
(2C)/128.5 (2C)/129.0 (d, Ar), 137.4/137.9 (s, Ar). FABMS m/z (%):
495 ([M+Na]+, 4), 55 (100). FABHRMS calcd for C26H32O8Na
(800 MHz, CDCl3) d: 1.370/1.374/1.39/1.45/1.50/1.55 [each s, 3H,
C(CH3)2], 3.32 (ddd, J = 10.3, 9.6, 4.8 Hz, 1H, H-50), 3.58 (dd, J = 9.6,
3.2 Hz, 1H, H-30), 3.67 (dd, J = 9.8, 6.2 Hz, 1H, H-7a), 3.68 (dd,
J = 8.0, 7.4 Hz, 1H, H-1a), 3.80 (s, 3H, OCH3), 3.89 (dd, J = 10.3,
10.3 Hz, 1H, H-60a), 3.95 (dd, J = 3.2, 0.6 Hz, 1H, H-20), 4.01 (dd,
J = 8.0, 6.2 Hz, 1H, H-1b), 4.03 (dd, J = 9.8, 6.2 Hz, 1H, H-7b), 4.17
(dd, J = 9.6, 9.6 Hz, 1H, H-40), 4.19 (dd, J = 6.2, 6.2 Hz, 1H, H-3),
4.26 (ddd, J = 7.4, 6.2, 6.2 Hz, 1H, H-2), 4.27 (dd, J = 6.2, 6.2 Hz, 1H,
H-4), 4.28 (dd, J = 10.3, 4.8 Hz, 1H, H-60b), 4.29 (dd, J = 6.2, 6.2 Hz,
1H, H-5), 4.38 (ddd, J = 6.2, 6.2, 6.2 Hz, 1H, H-6), 4.51 (d,
J = 0.6 Hz, 1H, H-10), 4.58/4.70 (each d, J = 12.4 Hz, 1H, OCH2Ph),
4.82 (s, 2H, OCH2C6H4OCH3), 5.61 (s, 1H, CHPh), 6.85 (d, J = 8.7 Hz,
2H, Ar), 7.26–7.31 (m, 5H, Ar), 7.35–7.39 (m, 5H, Ar), 7.49–7.50
(m, 2H, Ar). 13C NMR (200 MHz, CDCl3) d: 25.2/25.56/25.59/26.5/
26.9/27.4 [C(CH3)2], 55.2 (OCH3), 66.2 (C-1), 67.6 (C-50), 68.5 (C-
60), 68.6 (C-7), 72.5 (OCH2Ph), 74.3 (OCH2C6H4OCH3), 74.7 (C-4),
75.0 (C-5), 75.41/74.44 (C-2 and C-20), 75.7 (C-6), 77.5 (C-3),
77.9 (C-30), 78.6 (C-40), 101.4 (CHPh), 101.9 (C-10), 109.09/109.15/
109.4 [C(CH3)2], 113.5 (2C)/126.0 (2C)/127.5 (2C)/127.6/128.2
(2C)/128.3 (2C)/128.9/129.9 (2C) (d, Ar), 130.4/137.5/138.3/
159.2 (s, Ar). FABMS m/z (%): 815 ([M+Na]+, 3), 121 (100).
FABHRMS calcd for C44H56O13Na [M+Na]+: 815.3619; found:
815.3624.
[M+Na]+: 495.1995; found: 495.2010.
24
17i: A colorless oil. [
a]
ꢀ7.6 (c 0.99, CHCl3). IR (neat) cmꢀ1
:
D
3478, 1454, 1381, 1372, 1254, 1213, 1096, 1047. 1H NMR
(400 MHz, CDCl3) d: 0.05 [s, 6H, Si(CH3)2], 0.88 [s, 9H, SiC(CH3)3],
1.35/1.44 [each s, 3H, C(CH3)2], 2.61 (br s, 1H, OH), 3.34 (ddd,
J = 10.0, 9.6, 4.8 Hz, 1H, H-50), 3.58 (dd, J = 10.8, 5.2 Hz, 1H, H-1a),
3.61 (dd, J = 10.8, 7.6 Hz, 1H, H-1b), 3.656 (dd, J = 9.2, 3.2 Hz, 1H,
H-30), 3.67 (dd, J = 11.2, 8.8 Hz, 1H, H-4a), 3.89 (dd, J = 10.4,
10.0 Hz, 1H, H-60a), 4.13 (ddd, J = 7.6, 6.4, 5.2 Hz, 1H, H-2), 4.15
(dd, J = 9.6, 9.2 Hz, 1H, H-40), 4.23 (dd, J = 3.2, 0.8 Hz, 1H, H-20),
4.27 (dd, J = 11.2, 2.4 Hz, 1H, H-4b), 4.29 (dd, J = 10.4, 4.8 Hz, 1H,
H-60b), 4.44 (ddd, J = 8.8, 6.4, 2.4 Hz, 1H, H-3), 4.61 (d, J = 0.8 Hz,
1H, H-10), 4.78/4.85 (each d, J = 12.4 Hz, 1H, OCH2Ph), 5.61 (s, 1H,
3.1.10. Deprotection of PMB group at C20position in coupled
products (16g–16k)
To a mixture of 16g (368 mg, 0.577 mmol) CH2Cl2 (5.0 mL), and
H2O (200 lL) was added 2,3-dichloro-5,6-dicyano-p-benzoquinone

3768
N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
CHPh), 7.27–7.42 (m, 8H, Ar), 7.49–7.52 (m, 2H, Ar). 13C NMR
(100 MHz, CDCl3) d: ꢀ5.61/ꢀ5.58 [Si(CH3)2], 18.0 [SiC(CH3)3],
25.3/27.9 [C(CH3)2], 25.7 [3C, SiC(CH3)3], 61.5 (C-1), 66.9 (C-50),
68.5 (2C, C-4 and C-60), 69.6 (C-20), 72.3 (OCH2Ph), 76.47/76.49
(C-2 and C-3), 76.7 (C-30), 78.3 (C-40), 100.5 (C-10), 101.5 (CHPh),
108.8 [C(CH3)2], 126.0 (2C)/127.78/127.82 (2C)/128.2 (2C)/128.4
(2C)/129.0 (d, Ar), 137.4/137.9 (s, Ar). FABMS m/z (%): 639
([M+Na]+, 56), 91 (100). FABHRMS calcd for C33H48O9SiNa
CO(CH2)6CH3], 0.89 [s, 9H, SiC(CH3)3], 1.25ꢀ1.33 [m, 8H,
CO(CH2)2(CH2)4CH3], 1.66 [tt, J = 7.6, 7.6 Hz, 2H, COCH2CH2
(CH2)4CH3], 2.46 [t, J = 7.6 Hz, 2H, COCH2(CH2)5CH3], 3.38 (ddd,
J = 10.0, 10.0, 4.8 Hz, 1H, H-50), 3.68 (ddd, J = 10.0, 7.6, 4.0 Hz, 1H,
H-1a), 3.70 (dd, J = 10.0, 3.2 Hz, 1H, H-30), 3.73 (ddd, J =11.2, 4.0,
4.0 Hz, 1H, H-2a), 3.79 (ddd, J = 11.2, 7.6, 4.0 Hz, 1H, H-2b), 3.86
(ddd, J = 10.0, 4.0, 4.0 Hz, 1H, H-1b), 3.90 (dd, J = 10.0, 10.0 Hz,
1H, H-60a), 3.99 (dd, J = 10.0, 10.0 Hz, 1H, H-40), 4.33 (dd, J = 10.0,
4.8 Hz, 1H, H-60b), 4.62/4.74 (each d, J = 12.4 Hz, 1H, OCH2Ph),
4.73 (d, J = 1.2 Hz, 1H, H-10), 5.61 (s, 1H, CHPh), 5.69 (dd, J = 3.2,
1.2 Hz, 1H, H-20), 7.24–7.40 (m, 8H, Ar), 7.49–7.52 (m, 2H, Ar).
13C NMR (100 MHz, CDCl3) d: ꢀ5.4/ꢀ5.3 [Si(CH3)2], 14.1
[CO(CH2)6CH3], 18.3 [SiC(CH3)3], 22.6 [CO(CH2)5CH2CH3], 25.0
[COCH2CH2(CH2)5CH3], 25.9 [3C, SiC(CH3)3], 28.95/28.99
[CO(CH2)2(CH2)2(CH2)2CH3], 31.7 [CO(CH2)4CH2CH2CH3], 34.1
[COCH2(CH2)5CH3], 62.7 (C-2), 67.3 (C-50), 68.47 (C-20), 68.53 (C-
60), 71.2 (C-1), 71.6 (OCH2Ph), 75.8 (C-30), 78.0 (C-40), 100.1 (C-10),
101.5 (CHPh), 126.1 (2C)/127.7/127.8 (2C)/128.2 (2C)/128.3 (2C)/
128.9 (d, Ar), 137.4/137.7 (s, Ar), 173.1 (OCOCH2). FABMS m/z
(%): 665 ([M+Na]+, 2), 91 (100). FABHRMS calcd for C36H54O8SiNa
[M+Na]+: 665.3486; found: 665.3501.
[M+Na]+: 639.2965; found: 639.2973.
ꢀ13.4 (c 1.19, CHCl3). IR (neat) cmꢀ1
:
23
17j: A colorless oil. [
a]
D
3483, 1454, 1373, 1250, 1215, 1092, 1045. 1H NMR (400 MHz,
CDCl3) d: 1.37/1.38/1.46/1.48 [each s, 3H, C(CH3)2], 3.35 (ddd,
J = 10.0, 9.6, 4.8 Hz, 1H, H-50), 3.62 (dd, J = 10.0, 6.4 Hz, 1H, H-5a),
3.65 (dd, J = 9.6, 3.2 Hz, 1H, H-30), 3.70 (dd, J = 7.6, 6.0 Hz, 1H, H-
1a), 3.87 (dd, J = 10.4, 10.0 Hz, 1H, H-60a), 3.97 (dd, J = 10.0,
6.4 Hz, 1H, H-5b), 4.12 (dd, J = 9.6, 9.6 Hz, 1H, H-40), 4.13 (dd,
J = 6.4, 6.4 Hz, 1H, H-3), 4.14 (d, J = 3.2 Hz, 1H, H-20), 4.19 (dd,
J = 7.6, 6.8 Hz, 1H, H-1b), 4.23 (ddd, J = 6.8, 6.4, 6.0 Hz, 1H, H-2),
4.30 (ddd, J = 6.4, 6.4, 6.4 Hz, 1H, H-4), 4.33 (dd, J = 10.4, 4.8 Hz,
1H, H-60b), 4.56 (s, 1H, H-10), 4.77/4.85 (each d, J = 12.0 Hz, 1H,
OCH2Ph), 5.61 (s, 1H, CHPh), 7.28–7.42 (m, 8H, Ar), 7.48–7.51 (m,
2H, Ar). 13C NMR (100 MHz, CDCl3) d: 25.2/25.4/26.6/27.7
[C(CH3)2], 66.3 (C-1), 67.0 (C-50), 68.0 (C-5), 68.6 (C-60), 69.7 (C-
20), 72.5 (OCH2Ph), 74.1 (C-2), 75.4 (C-4), 76.6 (C-30), 77.9 (C-3),
78.3 (C-40), 100.5 (C-10), 101.6 (CHPh), 109.3/109.7 [C(CH3)2],
126.0 (2C)/127.8 (2C)/127.9/128.2 (2C)/128.5 (2C)/129.0 (d, Ar),
137.3/137.8 (s, Ar). FABMS m/z (%): 595 ([M+Na]+, 20), 154 (100).
Following the method described above, 17h (474 mg,
0.997 mmol), 17i (91.0 mg, 0.148 mmol), 17j (417 mg,
0.728 mmol), and 17k (150 mg, 0.223 mmol) were converted to
1,2-O-isopropylidene-
dene-2-O-octanoyl-b-
1-O-(tert-butyldimethylsilyl)-2,3-O-isopropylidene-
yl 3-O-benzyl-4,6-O-benzylidene-2-O-octanoyl-b- -mannopyrano-
side (18i, 97.0 mg, 88%), 1,2:3,4-di-O-isopropylidene- -arabinitol-
5-yl 3-O-benzyl-4,6-O-benzylidene-2-O-octanoyl-b- -mannopyra-
noside (18j, 509 mg, quant.), and 2,3:4,5:6,7-tri-O-iso-
D
-glycerol-3-yl
-mannopyranoside (18h, 565 mg, 95%),
-erythritol-4-
3-O-benzyl-4,6-O-benzyli-
D
D
FABHRMS calcd for
595.2512.
C
31H40O10Na [M+Na]+: 595.2519; found:
D
D
ꢀ7.4 (c 0.38, CHCl3). IR (neat) cmꢀ1
:
D
25
17k: A colorless oil. [
a]
D
3483, 1454, 1377, 1246, 1215, 1092, 1049. 1H NMR (800 MHz,
CDCl3) d: 1.375/1.384/1.39/1.45/1.48/1.55 [each s, 3H, C(CH3)2],
3.34 (ddd, J = 10.3, 9.6, 4.8 Hz, 1H, H-50), 3.60 (dd, J = 9.8, 5.5 Hz,
1H, H-7a), 3.65 (dd, J = 9.6, 3.4 Hz, 1H, H-30), 3.79 (dd, J = 8.0,
7.8 Hz, 1H, H-1a), 3.85 (dd, J = 10.3, 10.3 Hz, 1H, H-60a), 4.01 (dd,
J = 9.8, 7.6 Hz, 1H, H-7b), 4.04 (dd, J = 8.0, 6.6 Hz, 1H, H-1b), 4.09
(dd, J = 9.6, 9.6 Hz, 1H, H-40), 4.11 (dd, J = 3.4, 0.9 Hz, 1H, H-20),
4.23 (ddd, J = 7.8, 6.6, 4.1 Hz, 1H, H-2), 4.326 (dd, J = 6.9, 6.6 Hz,
1H, H-4), 4.328 (dd, J = 10.3, 4.8 Hz, 1H, H-60b), 4.37 (ddd, J = 7.6,
6.0, 5.5 Hz, 1H, H-6), 4.39 (dd, J = 6.9, 4.1 Hz, 1H, H-3), 4.41 (dd,
J = 6.6, 6.0 Hz, 1H, H-5), 4.55 (d, J = 0.9 Hz, 1H, H-10), 4.76/4.86 (each
d, J = 12.4 Hz, 1H, OCH2Ph), 5.60 (s, 1H, CHPh), 7.29–7.40 (m, 8H,
Ar), 7.49 (m, 2H, Ar). 13C NMR (200 MHz, CDCl3) d: 25.1/25.6/
25.8/26.4/26.8/27.8 [C(CH3)2], 66.4 (C-1), 66.9 (C-50), 68.1 (C-7),
68.6 (C-60), 69.7 (C-20), 72.6 (OCH2Ph), 75.20/75. 21 (C-4 and C-5),
75.3 (C-2), 75.7 (C-6), 76.3 (C-3), 76.6 (C-30), 78.4 (C-40), 100.4 (C-
10), 101.6 (CHPh), 108.8/109.0/109.6 [C(CH3)2], 126.0 (2C)/127.9
(2C)/128.0/128.3 (2C)/128.5 (2C)/129.0 (d, Ar), 137.3/137.7 (s, Ar).
FABMS m/z (%): 695 ([M+Na]+, 36), 91 (100). FABHRMS calcd for
C36H48O12Na [M+Na]+: 695.3043; found: 695.3030.
propylidene-
D
-glycero-
D
-galactoheptitol-1-yl
3-O-benzyl-4,6-O-
benzylidene-2-O-octanoyl-b-
D-mannopyranoside (18k, 160 mg,
90%), respectively.
23
18h: A colorless oil. [
a
]
ꢀ66.3 (c 1.31, CHCl3). IR (neat) cmꢀ1
:
D
1744, 1454, 1373, 1250, 1215, 1157, 1096, 1053. 1H NMR
(400 MHz, CDCl3) d: 0.87 [t, J = 7.2 Hz, 3H, CO(CH2)6CH3], 1.25–
1.34 [m, 8H, CO(CH2)2(CH2)4CH3], 1.35/1.40 [each s, 3H, C(CH3)2],
1.66 [tt, J = 7.2, 7.2 Hz, 2H, COCH2CH2(CH2)4CH3], 2.45 [t, J = 7.2 Hz,
2H, COCH2(CH2)5CH3], 3.38 (ddd, J = 10.0, 9.6, 4.8 Hz, 1H, H-50),
3.66 (dd, J = 10.4, 6.0 Hz, 1H, H-3a), 3.71 (dd, J = 8.4, 6.0 Hz, 1H, H-
1a), 3.72 (dd, J = 9.6, 3.6 Hz, 1H, H-30), 3.79 (dd, J = 10.4, 5.6 Hz, 1H,
H-3b), 3.90 (dd, J = 10.4, 10.0 Hz, 1H, H-60a), 3.99 (dd, J = 9.6,
9.6 Hz, 1H, H-40), 4.03 (dd, J = 8.4, 6.4 Hz, 1H, H-1b), 4.25 (dddd,
J = 6.4, 6.0, 6.0, 5.6 Hz, 1H, H-20), 4.33 (dd, J = 10.4, 4.8 Hz, 1H, H-
60b), 4.63/4.74 (each d, J = 12.4 Hz, 1H, OCH2Ph), 4.70 (d, J = 1.2 Hz,
1H, H-10), 5.61 (s 1H, CHPh), 5.67 (dd, J = 3.6, 1.2 Hz, 1H, H-20),
7.27–7.41 (m, 8H, Ar), 7.49–7.51 (m, 2H, Ar). 13C NMR (100 MHz,
CDCl3) d: 14.1 [CO(CH2)6CH3], 22.6 [CO(CH2)5CH2CH3], 25.0
[COCH2CH2(CH2)5CH3],
25.4/26.8
[C(CH3)2],
28.9/29.0
[CO(CH2)2(CH2)2(CH2)2CH3], 31.7 [CO(CH2)4CH2CH2CH3], 34.1
[COCH2(CH2)5CH3], 66.8 (C-1), 67.3 (C-50), 68.3 (C-20), 68.5 (C-60),
70.5, (C-3) 71.6 (OCH2Ph), 74.5 (C-2), 75.6 (C-30), 77.9 (C-40), 99.6
(C-10), 101.5 (CHPh), 109.6 [C(CH3)2], 126.0 (2C)/127.7 (3C)/128.2
(2C)/128.3 (2C)/129.0 (d, Ar), 137.3/137.7 (s, Ar), 173.2 (OCOCH2).
FABMS m/z (%): 621 ([M+Na]+, 20), 73 (100). HRMSFAB calcd for
3.1.11. Esterification of monoalcohols (17g–17k)
To a solution of 17g (216 mg, 0.418 mmol) and N,N-dimethy-
laminopyridine (DMAP, 153 mg, 1.25 mmol) in
a mixture of
CH2Cl2 (4.2 mL) and pyridine (0.14 mL, 1.67 mmol) was added n-
octanoyl chloride (0.36 mL, 2.09 mmol), and the mixture was stir-
red at room temperature for 7 h. After the reaction was quenched
with water, the resulting mixture was extracted with CH2Cl2. The
extract was successively washed with aqueous sodium bicarbon-
ate, water and brine, and concentrated in vacuo. The residue was
purified by column chromatography (n-hexane–EtOAc, 20/1) to
give 2-(tert-butyldimethylsiloxy)ethanol-1-yl 3-O-benzyl-4,6-O-
C
34H46O9Na [M+Na]+: 621.3040; found: 621.3039.
24
18i: A colorless oil. [
a]
ꢀ43.8 (c 1.29, CHCl3). IR (neat) cmꢀ1
:
D
1744, 1458, 1377, 1254, 1215, 1157, 1096, 1049. 1H NMR
(700 MHz, CDCl3) d: 0.047/0.048 [each s, 3H, Si(CH3)2], 0.87 [t,
J = 7.4 Hz, 3H, CO(CH2)6CH3], 0.88 [s, 9H, SiC(CH3)3], 1.22–1.31
[m, 8H, CO(CH2)2(CH2)4CH3], 1.35/1.42 [each s, 3H, C(CH3)2], 1.67
[tt, J = 7.6, 7.6 Hz, 2H, COCH2CH2(CH2)4CH3], 2.46 [t, J = 7.6 Hz,
2H, COCH2(CH2)5CH3], 3.38 (ddd, J = 10.0, 9.6, 4.8 Hz, 1H, H-50),
3.57 (dd, J = 10.8, 4.8 Hz, 1H, H-1a), 3.62 (dd, J = 10.8, 7.2 Hz, 1H,
H-1b), 3.70 (dd, J = 11.4, 8.2 Hz, 1H, H-4a), 3.73 (dd, J = 9.6,
benzylidene-2-O-octanoyl-b-D-mannopyranoside (18g, 207 mg,
25
77%) as a colorless oil. [
a]
ꢀ50.8 (c 1.08, CHCl3). IR (neat) cmꢀ1
:
D
1746, 1456, 1373, 1256, 1157, 1096. 1H NMR (400 MHz, CDCl3) d:
0.05/0.06 [each s, 3H, Si(CH3)2], 0.87 [t, J = 7.2 Hz, 3H,

N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
3769
3.4 Hz, 1H, H-30), 3.89 (dd, J = 10.4, 10.0 Hz, 1H, H-60a), 3.98 (dd,
J = 9.6, 9.6 Hz, 1H, H-40), 4.11 (ddd, J = 7.2, 6.4, 4.8 Hz, 1H, H-2),
4.14 (dd, J = 11.4, 3.0 Hz, 1H, H-4b), 4.31 (dd, J = 10.4, 4.8 Hz, 1H,
H-60b), 4.34 (ddd, J = 8.2, 6.4, 3.0 Hz, 1H, H-3), 4.62/4.745 (each d,
J = 12.4 Hz, 1H, OCH2Ph), 4.744 (d, J = 1.2 Hz, 1H, H-10), 5.61 (s,
1H, CHPh), 5.72 (dd, J = 3.4, 1.2 Hz, 1H, H-20), 7.26–7.32 (m, 3H,
Ar), 7.36–7.40 (m, 5H, Ar), 7.49–7.51 (m, 2H, Ar). 13C NMR
(175 MHz, CDCl3) d: ꢀ5.52/ꢀ5.49 [Si(CH3)2], 14.0 [CO(CH2)6CH3],
18.1 [SiC(CH3)3], 22.6 [CO(CH2)5CH2CH3], 25.0 [COCH2CH2
(CH2)5CH3], 25.3/27.9 [C(CH3)2], 25.8 [3C, SiC(CH3)3], 28.9/29.0
[CO(CH2)2(CH2)2(CH2)2CH3], 31.7 [CO(CH2)4CH2CH2CH3], 34.2
[COCH2(CH2)5CH3], 61.6 (C-1), 67.3 (C-50), 68.4 (C-4), 68.49 (C-20),
68.52 (C-60), 71.5 (OCH2Ph), 75.7 (C-30), 76.78 (C-3), 76.82 (C-2),
78.0 (C-40), 99.7 (C-10), 101.5 (CHPh), 108.7 [C(CH3)2], 126.1 (2C)/
127.67/127.72 (2C)/128.2 (2C)/128.3 (2C)/128.9 (d, Ar), 137.4/
137.7 (s, Ar), 173.2 (OCOCH2). FABMS m/z (%): 765 ([M+Na]+, 1),
73 (100). FABHRMS calcd for C41H62O10SiNa [M+Na]+: 765.4010;
(CH2)2CH3], 31.7 [CO(CH2)4CH2CH2CH3], 34.1 [COCH2(CH2)5CH3],
66.3 (C-1), 67.5 (C-50), 68.3/68.4 (C-7 and C-60), 68.4 (C-20), 71.7
(OCH2Ph), 74.8/75.1/75.2/75.3 (C-2, C-4, C-5 and C-6), 75.7 (C-30),
76.8 (C-3), 77.9 (C-40), 99.8 (C-10), 101.6 (CHPh), 109.0/109.2/
109.5 [C(CH3)2], 126.0 (2C)/127.69 (2C)/127.72/128.2 (2C)/128.3
(2C)/129.0 (d, Ar), 137.2/137.6 (s, Ar), 173.1 (OCOCH2). FABMS m/
z
C
(%): 821 ([M+Na]+, 4), 91 (100). FABHRMS calcd for
44H62O13Na [M+Na]+: 821.4088; found: 821.4090.
3.1.12. Hydrogenolysis of octanoyl esters (18g–18k)
A suspension of 18g (207 mg, 0.322 mmol) and 10% palladium on
carbon (20 mg) in EtOAc (3.2 mL) was hydrogenated at room tem-
perature until the uptake of hydrogen ceased. The catalyst was fil-
tered off and washed with MeOH. The combined filtrate and
washings were condensed in vacuo. The residue was purified by col-
umn chromatography (CHCl3–MeOH, 50/1) to give 2-(tert-
butyldimethylsiloxy)ethanol-1-yl 2-O-octanoyl-b-
D
-mannopyra-
25
found: 765.3992.
noside (19g, 116 mg, 78%) as a colorless oil. [a]
ꢀ29.0 (c 1.02,
D
24
18j: A colorless oil. [
a]
ꢀ53.9 (c 1.55, CHCl3). IR (neat) cmꢀ1
:
CHCl3). IR (neat) cmꢀ1: 3383, 1748, 1472, 1362, 1256, 1161, 1074.
1H NMR (400 MHz, CD3OD) d: 0.067/0.071 [each s, 3H, Si(CH3)2],
0.90 [s, 9H, SiC(CH3)3], 0.91 [t, J = 7.2 Hz, 3H, CO(CH2)6CH3], 1.27–
1.39 [m, 8H, CO(CH2)2(CH2)4CH3], 1.63 [ddt, J = 7.6, 7.6, 7.6 Hz, 2H,
COCH2CH2(CH2)4CH3], 2.35/2.40 [each dt, J = 15.6, 7.6 Hz, 1H,
COCH2(CH2)5CH3], 3.26 (ddd, J = 9.6, 6.4, 2.4 Hz, 1H, H-50), 3.50
(dd, J = 9.6, 9.6 Hz, 1H, H-40), 3.63 (dd, J = 9.6, 3.6 Hz, 1H, H-30),
3.64 (ddd, J = 10.8, 6.8, 4.4 Hz, 1H, H-1a), 3.70 (dd, J = 11.6, 6.4 Hz,
1H, H-60a), 3.75ꢀ3.80 (m, 2H, H-2a and H-2b), 3.88 (ddd, J = 10.8,
4.8, 4.0 Hz, 1H, H-1b), 3.90 (dd, J = 11.6, 2.4 Hz, 1H, H-60b), 4.71 (d,
J = 0.8 Hz, 1H, H-10), 5.36 (dd, J = 3.6, 0.8 Hz, 1H, H-20). 13C NMR
(100 MHz, CD3OD) d: ꢀ5.2/ꢀ5.1 [Si(CH3)2], 14.4 [CO(CH2)6CH3], 19.2
[SiC(CH3)3], 23.7 [CO(CH2)5CH2CH3], 26.0 [COCH2CH2(CH2)5CH3],
26.4 [3C, SiC(CH3)3], 30.1/30.2 [CO(CH2)2(CH2)2(CH2)2CH3], 32.9
[CO(CH2)4CH2CH2CH3], 35.1 [COCH2(CH2)5CH3], 62.9 (C-60), 63.9
(C-2), 68.9 (C-40), 71.9 (C-1), 72.9 (C-20), 73.7 (C-30), 78.6 (C-50), 100.7
(C-10), 175.0 (OCOCH2). FABMS m/z (%): 487 ([M+Na]+, 100). FABHRMS
calcd for C22H44O8SiNa [M+Na]+: 487.2703; found: 487.2723.
D
1744, 1454, 1373, 1250, 1219, 1157, 1092, 1049. 1H NMR
(400 MHz, CDCl3) d: 0.87 [t, J = 6.8 Hz, 3H, CO(CH2)6CH3], 1.24–
1.38 [m, 8H, CO(CH2)2(CH2)4CH3], 1.37 [s, 6H, C(CH3)2], 1.456/
1.462 [each s, 3H, C(CH3)2], 1.67 [tt, J = 7.6, 7.6 Hz, 2H,
COCH2CH2(CH2)4CH3], 2.43/2.48 [each dt, J = 16.0, 7.6 Hz, 1H,
COCH2(CH2)5CH3], 3.37 (ddd, J = 10.0, 10.0, 4.8 Hz, 1H, H-50), 3.56
(dd, J = 10.0, 4.8 Hz, 1H, H-5a), 3.61 (dd, J = 8.4, 7.2 Hz, 1H, H-1a),
3.71 (dd, J = 10.0, 3.2 Hz, 1H, H-30), 3.87 (dd, J = 10.4, 10.0 Hz, 1H,
H-60a), 3.93 (dd, J = 10.0, 8.0 Hz, 1H, H-5b), 3.96 (dd, J = 10.0,
10.0 Hz, 1H, H-40), 4.09 (dd, J = 6.4, 5.6 Hz, 1H, H-3), 4.12 (dd,
J = 8.4, 6.8 Hz, 1H, H-1b), 4.19 (ddd, J = 8.0, 5.6, 4.8 Hz, 1H, H-4),
4.25 (ddd, J = 7.2, 6.8, 6.4 Hz, 1H, H-2), 4.34 (dd, J = 10.4, 4.8 Hz,
1H, H-60b), 4.62/4.73 (each d, J = 12.4 Hz, 1H, OCH2Ph), 4.64 (d,
J = 0.8 Hz, 1H, H-10), 5.61 (s, 1H, CHPh), 5.65 (dd, J = 3.2, 0.8 Hz,
1H, H-20), 7.25–7.42 (m, 8H, Ar), 7.49–7.51 (m, 2H, Ar). 13C NMR
(100 MHz, CDCl3) d: 14.1 [CO(CH2)6CH3], 22.6 [CO(CH2)5CH2CH3],
24.9 [COCH2CH2(CH2)5CH3], 25.2/25.4/26.6/27.8 [C(CH3)2], 28.96/
29.00 [CO(CH2)2(CH2)2(CH2)2CH3], 31.7 [CO(CH2)4CH2CH2CH3],
34.2 [COCH2(CH2)5CH3], 66.4 (C-1), 67.4 (C-50), 67.8 (C-5), 68.4
(C-20), 68.5 (C-60), 71.6 (OCH2Ph), 74.1 (C-2), 74.9 (C-4), 75.6 (C-
30), 77.9 (C-40), 78.4 (C-3), 99.6 (C-10), 101.6 (CHPh), 109.2/109.5
[C(CH3)2], 126.1 (2C)/127.7 (3C)/128.2 (2C)/128.3 (2C)/129.0 (d,
Ar), 137.3/137.6 (s, Ar), 173.1 (OCOCH2). FABMS m/z (%): 721
([M+Na]+, 2), 91 (100). FABHRMS calcd for C39H54O11Na [M+Na]+:
Following the method described above, 18h (107 mg,
0.179 mmol), 18i (84.9 mg, 0.114 mmol), 18j (454 mg,
0.650 mmol), and 18k (150 mg, 0.188 mmol) were converted to
1,2-O-isopropylidene-
anoside (19h, 55.9 mg, 74%), 1-O-(tert-butyldimethylsilyl)-2,3-O-
isopropylidene- -erythritol-4-yl 2-O-octanoyl-b- -mannopyrano-
side (19i, 47.2 mg, 73%), 1,2:3,4-di-O-isopropylidene- -arabinitol-
5-yl 2-O-octanoyl-b- -mannopyranoside (19j, 294 mg, 87%), and
2,3:4,5:6,7-tri-O-isopropylidene- -glycero- -galactoheptitol-1-yl
-mannopyranoside (19k, 81.0 mg, 70%),
D-glycerol-3-yl 2-O-octanoyl-b-D-mannopyr-
D
D
D
721.3564; found: 721.3592.
D
25
18k: A colorless amorphous. [
a]
ꢀ42.9 (c 1.07, CHCl3). IR
D
D
D
(neat) cmꢀ1: 1744, 1454, 1377, 1246, 1215, 1157, 1092, 1049. 1H
NMR (800 MHz, CDCl3) d: 0.87 [t, J = 7.2 Hz, 3H, CO(CH2)6CH3],
1.22–1.39 [m, 8H, CO(CH2)2(CH2)4CH3], 1.37 [s, 6H, C(CH3)2],
1.38/1.45/1.47/1.55 [each s, 3H, C(CH3)2], 1.61–1.69 [m, 2H,
COCH2CH2(CH2)4CH3], 2.41/ 2.45 [each ddd, J = 15.8, 8.2, 6.9 Hz,
1H, COCH2(CH2)5CH3], 3.38 (ddd, J = 9.8, 9.8, 4.8 Hz, 1H, H-50),
3.61 (dd, J = 10.5, 5.3 Hz, 1H, H-7a), 3.71 (dd, J = 9.8, 3.2 Hz, 1H,
H-30), 3.77 (dd, J = 8.0, 7.3 Hz, 1H, H-1a), 3.87 (dd, J = 10.6, 9.8 Hz,
1H, H-60a), 3.96 (dd, J = 9.8, 9.8 Hz, 1H, H-40), 3.99 (dd, J = 10.5,
8.0 Hz, 1H, H-7b), 4.08 (dd, J = 8.0, 6.4 Hz, 1H, H-1b), 4.11 (dd,
J = 6.7, 4.6 Hz, 1H, H-3), 4.16 (ddd, J = 7.3, 6.4, 4.6 Hz, 1H, H-2),
4.3057 (dd, J = 6.7, 6.0 Hz, 1H, H-4), 4.3061 (ddd, J = 8.0, 6.0,
5.3 Hz, 1H, H-6), 4.32 (dd, J = 10.6, 4.8 Hz, 1H, H-60b), 4.35 (dd,
J = 6.0, 6.0 Hz, 1H, H-5), 4.62/4.72 (each d, J = 12.3 Hz, 1H,
OCH2Ph), 4.66 (d, J = 1.2 Hz, 1H, H-10), 5.61 (s, 1H, CHPh), 5.63
(dd, J = 3.2, 1.2 Hz, 1H, H-20), 7.26–7.28 (m, 1H, Ar), 7.30–7.32 (m,
2H, Ar), 7.34–7.35 (m, 2H, Ar), 7.36–7.40 (m, 3H, Ar), 7.49–7.50
(m, 2H, Ar). 13C NMR (100 MHz, CDCl3) d: 14.1 [CO(CH2)6CH3],
22.6 [CO(CH2)5CH2CH3], 24.9 [COCH2CH2(CH2)5CH3], 25.1/25.6/
2-O-octanoyl-b-D
respectively.
24
19h: A colorless oil. [
a]
ꢀ42.0 (c 0.79, CHCl3). IR (neat) cmꢀ1
:
D
3383, 1732, 1454, 1416, 1373, 1261, 1219, 1165, 1072. 1H NMR
(400 MHz, CDCl3) d: 0.88 [t, J = 7.2 Hz, 3H, CO(CH2)6CH3], 1.25–
1.35 [m, 8H, CO(CH2)2(CH2)4CH3], 1.35/1.40 [each s, 3H, C(CH3)2],
1.63 [tt, J = 7.6, 7.6 Hz, 2H, COCH2CH2(CH2)4CH3], 2.41 [t,
J = 7.6 Hz, 2H, COCH2(CH2)5CH3], 3.34 (ddd, J = 8.0, 4.8, 3.2 Hz, 1H,
H-50), 3.64 (dd, J = 10.4, 5.6 Hz, 1H, H-3a), 3.72–3.76 (m, 2H, H-30
and H-40), 3.73 (dd, J = 8.0, 6.0 Hz, 1H, H-1a), 3.81 (dd, J = 10.4,
5.6 Hz, 1H, H-3b), 3.86 (dd, J = 12.0, 4.8 Hz, 1H, H-60a), 3.94 (dd,
J = 12.0, 3.2 Hz, 1H, H-60b), 4.02 (dd, J = 8.0, 6.0 Hz, 1H, H-1b),
4.25 (dddd, J = 6.0, 6.0, 5.6, 5.6 Hz, 1H, H-2), 4.71 (s, 1H, H-10),
5.39 (br s, 1H, H-20). 13C NMR (100 MHz, CDCl3) d: 14.0
[CO(CH2)6CH3], 22.6 [CO(CH2)5CH2CH3], 24.9 [COCH2CH2
(CH2)5CH3], 25.4/26.7 [C(CH3)2], 28.9/29.0 [CO(CH2)2(CH2)2
(CH2)2CH3], 31.7 [CO(CH2)4CH2CH2CH3], 34.2 [COCH2(CH2)5CH3],
62.4 (C-60), 66.6 (C-1), 68.4 (C-40), 70.2 (C-3), 70.9 (C-20), 72.9 (C-
30), 74.5 (C-2), 75.7 (C-50), 98.9 (C-10), 109.6 [C(CH3)2], 174.2
(OCOCH2). FABMS m/z (%): 443 ([M+Na]+, 40), 57 (100).
25.7/26.5/26.9/27.6
[C(CH3)2],
28.9/29.0
[CO(CH2)2(CH2)2

3770
N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
FABHRMS calcd for C20H36O9Na [M+Na]+: 443.2257; found:
[COCH2(CH2)5CH3], 63.0 (C-60), 67.4 (C-1), 69.0 (C-40), 69.1 (C-7),
72.9 (C-20), 73.6 (C-30), 76.3/76.46/76.50/76.8 (C-2, C-4, C-5 and
C-6), 77.6 (C-3), 78.8 (C-50), 100.7 (C-10), 109.8/110.1/110.6
[C(CH3)2], 175.0 (OCOCH2). FABMS m/z (%): 643 ([M+Na]+, 99),
127 (100). FABHRMS calcd for C30H52O13Na [M+Na]+: 643.3306;
found: 643.3305.
443.2260.
23
19i: A colorless oil. [
a]
ꢀ22.2 (c 1.11, CHCl3). IR (neat) cmꢀ1
:
D
3399, 1744, 1458, 1377, 1254, 1219, 1165, 1076. 1H NMR
(400 MHz, CD3OD) d: 0.085/0.087 [each s, 3H, Si(CH3)2], 0.90 [t,
J = 7.2 Hz, 3H, CO(CH2)6CH3], 0.91 [s, 9H, SiC(CH3)3], 1.27–1.39
[m, 8H, CO(CH2)2(CH2)4CH3], 1.32/1.40 [each s, 3H, C(CH3)2], 1.62
[ddt, J = 7.6, 7.6, 7.6 Hz, 2H, COCH2CH2(CH2)4CH3], 2.35/2.39 [each
dt, J = 16.0, 7.6 Hz, 1H, COCH2(CH2)5CH3], 3.24 (ddd, J = 9.6, 6.0,
2.4 Hz, 1H, H-50), 3.51 (dd, J = 9.6, 9.6 Hz, 1H, H-40), 3.63 (dd,
J = 9.6, 3.2 Hz, 1H, H-30), 3.65 (dd, J = 11.2, 6.0 Hz, 1H, H-1a), 3.66
(dd, J = 11.2, 6.8 Hz, 1H, H-4a), 3.71 (dd, J = 12.0, 6.0 Hz, 1H, H-
60a), 3.75 (dd, J = 11.2, 5.2 Hz, 1H, H-1b), 3.89 (dd, J = 12.0, 2.4 Hz,
1H, H-60b), 4.01 (dd, J = 11.2, 4.8 Hz, 1H, H-4b), 4.16 (ddd, J = 6.4,
6.0, 5.2 Hz, 1H, H-2), 4.29 (ddd, J = 6.8, 6.4, 4.8 Hz, 1H, H-3), 4.70
(d, J = 0.8 Hz, 1H, H-10), 5.35 (dd, J = 3.2, 0.8 Hz, 1H, H-20).
13C NMR (100 MHz, CD3OD) d: ꢀ5.2/ꢀ5.1 [Si(CH3)2], 14.5
[CO(CH2)6CH3], 19.1 [SiC(CH3)3], 23.7 [CO(CH2)5CH2CH3], 25.5/
28.1 [C(CH3)2], 26.0 [COCH2CH2(CH2)5CH3], 26.4 [3C, SiC(CH3)3],
30.1/30.2 [CO(CH2)2(CH2)2(CH2)2CH3], 32.9 [CO(CH2)4CH2CH2CH3],
35.1 [COCH2(CH2)5CH3], 62.9 (C-60), 63.2 (C-1), 68.9 (C-40), 69.0
(C-4), 72.9 (C-20), 73.6 (C-30), 77.4 (C-3), 78.5 (C-50), 79.0 (C-2),
100.4 (C-10), 109.8 [C(CH3)2], 175.0 (OCOCH2). FABMS m/z (%):
587 ([M+Na]+, 20), 73 (100). FABHRMS calcd for C27H52O10SiNa
3.1.13. Esterification of trisalcohols (19g–19k)
To a solution of 19g (103 mg, 0.221 mmol) and DMAP (80.9 mg,
0.662 mmol) in
a mixture of CH2Cl2 (2.2 mL) and pyridine
(0.07 mL, 0.883 mmol) was added hexanoic anhydride (0.20 mL,
0.883 mmol) at room temperature, and the reaction mixture was
stirred at room temperature for 30 min. After the reaction was
quenched with water, the resulting mixture was extracted with
CH2Cl2. The extract was successively washed with aqueous sodium
bicarbonate, water and brine, and condensed in vacuo. The residue
was purified by column chromatography (n-hexane–EtOAc, 30/1)
to give 2-(tert-butyldimethylsiloxy)ethanol-1-yl 3,4,6-tri-O-hex-
anoyl-2-O-octanoyl-b-D-mannopyranoside (20g, 134 mg, 80%) as
25
a colorless oil. [a]
ꢀ25.4 (c 1.06, CHCl3). IR (neat) cmꢀ1: 1748,
D
1456, 1248, 1161, 1103, 1070. 1H NMR (400 MHz, CDCl3) d:
0.045/0.049 [each s, 3H, Si(CH3)2], 0.86–0.91 (m, 12H, acyl CH3),
0.89 [s, 9H, SiC(CH3)3], 1.22–1.36 (m, 20H, acyl CH2), 1.50–1.67
(m, 8H, acyl CH2), 2.17/2.22/2.23/2.28/2.40/2.45 (dt, J = 16.0,
7.6 Hz, 1H, COCH2), 2.34 (t, J = 7.6 Hz, 2H, COCH2), 3.65 (ddd,
J = 10.0, 5.6, 2.4 Hz, 1H, H-50), 3.66 (ddd, J = 10.0, 7.6, 4.0 Hz, 1H,
H-1a), 3.73 (ddd, J = 11.2, 4.0, 4.0 Hz, 1H, H-2a), 3.79 (ddd,
J = 11.2, 7.6, 4.0 Hz, 1H, H-2b), 3.87 (ddd, J = 10.0, 4.0, 4.0 Hz, 1H,
H-1b), 4.17 (dd, J = 12.0, 2.4 Hz, 1H, H-60a), 4.25 (dd, J = 12.0,
5.6 Hz, 1H, H-60b), 4.77 (s, 1H, H-10), 5.05 (dd, J = 10.0, 3.6 Hz, 1H,
H-30), 5.26 (dd, J = 10.0, 10.0 Hz, 1H, H-40), 5.50 (d, J = 3.6 Hz, 1H,
H-20). 13C NMR (100 MHz, CDCl3) d: ꢀ5.4/ꢀ5.3 [Si(CH3)2], 13.8
(2C)/13.9/14.0 (acyl CH3), 18.3 [SiC(CH3)3], 22.2 (2C)/22.3/22.6
(acyl CH2), 24.2/24.4/24.5/25.0 (COCH2CH2), 25.8 [3C, SiC(CH3)3],
28.95/28.99 [CO(CH2)2(CH2)2(CH2)2CH3], 31.2 (2C)/31.3/31.7 (acyl
CH2), 33.9/34.0 (2C)/34.1 (COCH2), 62.45/62.53 (C-2 and C-60),
65.8 (C-40), 68.6 (C-20), 71.0 (C-30), 71.1 (C-1), 72.5 (C-50), 99.1 (C-
10), 172.3/172.6/173.0/173.5 (OCOCH2). FABMS m/z (%): 781
([M+Na]+, 4), 99 (100). FABHRMS calcd for C40H74O11SiNa
[M+Na]+: 781.4898; found: 781.4883.
[M+Na]+: 587.3227; found: 587.3250.
25
19j: A colorless oil. [
a]
ꢀ32.8 (c 0.51, CHCl3). IR (neat) cmꢀ1
:
D
3395, 1736, 1458, 1373, 1250, 1215, 1161, 1072. 1H NMR
(400 MHz, CDCl3) d: 0.88 [t, J = 7.2 Hz, 3H, CO(CH2)6CH3], 1.24–
1.38 [m, 8H, CO(CH2)2(CH2)4CH3], 1.37/1.38/1.45/1.46 [each s, 3H,
C(CH3)2], 1.64 [tt, J = 7.2, 7.2 Hz, 2H, COCH2CH2(CH2)4CH3], 2.39/
2.44 [each dt, J = 16.0, 7.2 Hz, 1H, COCH2(CH2)5CH3], 3.34 (ddd,
J = 8.8, 4.4, 3.6 Hz, 1H, H-50), 3.58 (dd, J = 10.0, 4.8 Hz, 1H, H-5a),
3.59 (dd, J = 8.0, 7.2 Hz, 1H, H-1a), 3.74 (dd, J = 9.6, 2.4 Hz, 1H, H-
30), 3.79 (dd, J = 9.2, 8.8 Hz, 1H, H-40), 3.87 (dd, J = 12.0, 4.4 Hz,
1H, H-60a), 3.92 (dd, J = 10.0, 8.0 Hz, 1H, H-5b), 3.94 (dd, J = 12.0,
3.6 Hz, 1H, H-60b), 4.10 (dd, J = 6.8, 6.0 Hz, 1H, H-3), 4.18 (dd,
J = 8.0, 6.0 Hz, 1H, H-1b), 4.16–4.21 (m, 1H, H-4), 4.24 (ddd,
J = 7.2, 6.8, 6.0 Hz, 1H, H-2), 4.67 (s, 1H, H-10), 5.38 (d, J = 2.4 Hz,
1H, H-20). 13C NMR (100 MHz, CDCl3) d: 14.0 [CO(CH2)6CH3], 22.6
[CO(CH2)5CH2CH3], 24.9 [COCH2CH2(CH2)5CH3], 25.4 (2C)/26.6/
27.7 [C(CH3)2], 28.9/29.0 [CO(CH2)2(CH2)2(CH2)2CH3], 31.7
[CO(CH2)4CH2CH2CH3], 34.2 [COCH2(CH2)5CH3], 62.3 (C-60), 66.4
(C-1), 67.8 (C-5), 68.2 (C-40), 70.9 (C-20), 73.0 (C-30), 74.3 (C-2),
75.0 (C-4), 75.7 (C-50), 78.2 (C-3), 98.8 (C-10), 109.2/109.6
[C(CH3)2], 174.2 (OCOCH2). FABMS m/z (%): 543 ([M+Na]+, 100).
Following the method described above, 19h (45.5 mg,
0.108 mmol), 19i (40.0 mg, 0.069 mmol), 19j (286 mg,
0.550 mmol), and 19k (70.0 mg, 0.113 mmol) were converted to
1,2-O-isopropylidene-
tanoyl-b- -mannopyranoside (20h, 77.0 mg, quant.), 1-O-(tert-
butyldimethylsilyl)-2,3-O-isopropylidene- -erythritol-4-yl 3,4,6-
tri-O-hexanoyl-2-O-octanoyl-b- -mannopyranoside (20i, 58.7 mg,
97%), 1,2:3,4-di-O-isopropylidene- -arabinitol-5-yl 3,4,6-tri-O-
hexanoyl-2-O-octanoyl-b- -mannopyranoside (20j, 351 mg, 78%),
and 2,3:4,5:6,7-tri-O-isopropylidene- -glycero- -galactoheptitol-
1-yl 3,4,6-tri-O-hexanoyl-2-O-octanoyl-b- -mannopyranoside
(20k, 95.0 mg, 92%), respectively.
D-glycerol-3-yl 3,4,6-tri-O-hexanoyl-2-O-oc-
D
FABHRMS calcd for
543.2778.
C
25H44O11Na [M+Na]+: 543.2781; found:
D
D
24
19k: A colorless amorphous. [
a]
ꢀ19.1 (c 1.01, CHCl3). IR
D
D
(neat) cmꢀ1: 3445, 1740, 1458,1377, 1250, 1215, 1157, 1069. 1H
NMR (400 MHz, CD3OD) d: 0.90 [t, J = 6.8 Hz, 3H, CO(CH2)6CH3],
1.26–1.38 [m, 8H, CO(CH2)2(CH2)4CH3], 1.33/1.34/1.35/1.39/1.42/
1.48 [each s, 3H, C(CH3)2], 1.62 [tdd, J = 7.6, 7.6, 7.6 Hz, 2H,
COCH2CH2(CH2)4CH3], 2.34/2.40 [each dt, J = 16.0, 7.6 Hz, 1H,
COCH2(CH2)5CH3], 3.28 (ddd, J = 9.6, 6.4, 2.0 Hz, 1H, H-50), 3.47
(dd, J = 9.6, 9.6 Hz, 1H, H-40), 3.58 (dd, J = 10.8, 4.8 Hz, 1H, H-7a),
3.63 (dd, J = 9.6, 3.2 Hz, 1H, H-30), 3.69 (dd, J = 12.0, 6.4 Hz, 1H,
H-60a), 3.77 (dd, J = 7.6, 7.6 Hz, 1H, H-1a), 3.91 (dd, J = 12.0,
2.0 Hz, 1H, H-60b), 4.00 (dd, J = 10.8, 8.4 Hz, 1H, H-7b), 4.06 (dd,
J = 7.6, 6.4 Hz, 1H, H-1b), 4,13 (dd, J = 6.4, 4.0 Hz, 1H, H-3), 4.17
(ddd, J = 7.6, 6.4, 4.0 Hz, 1H, H-2), 4.33 (ddd, J = 8.4, 5.6, 4.8 Hz,
1H, H-6), 4.37 (dd, J = 6.4, 6.4 Hz, 1H, H-4), 4.42 (dd, J = 6.4,
5.6 Hz, 1H, H-5), 4.71 (d, J = 0.8 Hz, 1H, H-10), 5.31 (dd, J = 3.2,
0.8 Hz, 1H, H-20). 13C NMR (100 MHz, CD3OD) d: 14.4
[CO(CH2)6CH3], 23.7 [CO(CH2)5CH2CH3], 25.6 [COCH2CH2
D
D
D
D
25
20h: A colorless oil. [
a]
ꢀ41.6 (c 1.05, CHCl3). IR (neat) cmꢀ1
:
D
1748, 1458, 1373, 1242, 1165, 1103, 1072. 1H NMR (400 MHz,
CDCl3) d: 0.86–0.92 (m, 12H, acyl CH3), 1.24–1.36 (m, 20H, acyl
CH2), 1.35/1.40 [each s, 3H, C(CH3)2], 1.50–1.68 (m, 8H, acyl CH2),
2.17/2.225/2.234/2.29/2.40/2.44 (each dt, J = 16.0, 7.2 Hz, 1H,
COCH2), 2.34 (t, J = 7.6 Hz, 2H, COCH2), 3.64–3.68 (m, 1H, H-50a),
3.66 (dd, J = 10.4, 6.0 Hz, 1H, H-3a), 3.69 (dd, J = 8.4, 6.0 Hz, 1H,
H-1a), 3.78 (dd, J = 10.4, 5.6 Hz, 1H, H-3b), 4.02 (dd, J = 8.4,
6.0 Hz, 1H, H-1b), 4.17 (dd, J = 12.0, 2.4 Hz, 1H, H-60a), 4.244 (dd,
J = 12.2, 5.6 Hz, 1H, H-60b), 4.244 (dddd, J = 6.0, 6.0, 6.0, 5.6 Hz,
1H, H-2), 4.77 (s, 1H, H-10), 5.06 (dd, J = 10.0, 3.2 Hz, 1H, H-30),
5.26 (dd, J = 10.0, 10.0 Hz, 1H, H-40), 5.50 (d, J = 3.2 Hz, 1H, H-20).
13C NMR (100 MHz, CDCl3) d: 13.8 (2C)/13.9/14.0 (acyl CH3), 22.2
(2C)/22.3/22.6 (acyl CH2), 24.3/24.4/24.5/25.0 (COCH2CH2), 25.3/
(CH2)5CH3],
25.8/25.9/26.0/26.8/27.2/27.9
[C(CH3)2],
30.2
[2C, CO(CH2)2(CH2)2(CH2)2CH3], 32.9 [CO(CH2)4CH2CH2CH3], 35.1

N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
3771
26.8 [C(CH3)2], 28.9/29.0 [CO(CH2)2(CH2)2(CH2)2CH3], 31.1 (2C)/
31.2/31.7 (acyl CH2), 33.9/34.0 (2C)/34.1 (COCH2), 62.4 (C-60),
65.8 (C-40), 66.7 (C-1), 68.4 (C-20), 70.4 (C-3), 70.9 (C-30), 72.6 (C-
50), 74.6 (C-2), 98.5 (C-10), 109.6 [C(CH3)2], 172.2/172.6/173.0/
173.4 (OCOCH2). FABMS m/z (%): 737 ([M+Na]+, 8), 99 (100).
4.36 (dd, J = 6.4, 5.6 Hz, 1H, H-5), 4.73 (d, J = 0.8 Hz, 1H, H-10), 5.04
(dd, J = 10.0, 3.2 Hz, 1H, H-30), 5.25 (dd, J = 10.0, 10.0 Hz, 1H, H-40),
5.47 (dd, J = 3.2, 0.8 Hz, 1H, H-20). 13C NMR (100 MHz, CDCl3) d:
13.8 (2C)/13.9/14.0 (acyl CH3), 22.2 (2C)/22.3/22.6 (acyl CH2), 24.2/
24.4/24.5/24.97
(COCH2CH2),
25.03/25.6/25.7/26.4/26.8/27.7
FABHRMS calcd for
737.4425.
C
38H66O12Na [M+Na]+: 737.4452; found:
[C(CH3)2], 28.95/29.03 [CO(CH2)2(CH2)2(CH2)2CH3], 31.2 (2C)/31.3/
31.7 (acyl CH2), 33.89/33.94/34.0/34.1 (COCH2), 62.3 (C-60), 65.5
(C-40), 66.2 (C-1), 68.2 (C-7), 68.4 (C-20), 71.0 (C-30), 72.7 (C-50),
74.8/75.1 (2C)/75.3 (C-2, C-4, C-5 and C-6), 76.6 (C-3), 98.7 (C-10),
109.0/109.2/109.6 [C(CH3)2], 172.2/172.6/172.9/173.4 (OCOCH2).
FABMS m/z (%): 937 ([M+Na]+, 3), 99 (100). FABHRMS calcd for
23
20i: A colorless oil. [
a
]
ꢀ17.4 (c 1.05, CHCl3). IR (neat) cmꢀ1
:
D
1748, 1466, 1377, 1250, 1219, 1165, 1099, 1072. 1H NMR
(400 MHz, CDCl3) d: 0.050/0.052 [each s, 3H, Si(CH3)2], 0.86–0.92
(m, 12H, acyl CH3), 0.88 [s, 9H, SiC(CH3)3], 1.20–1.36 (m, 20H, acyl
CH2), 1.34/1.43 [each s, 3H, C(CH3)2], 1.51–1.70 (m, 8H, acyl CH2),
2.17/2.22 (each dt, J = 16.0, 7.6 Hz, 1H, COCH2), 2.23/2.28/2.41/
2.45 (each dt, J = 15.6, 7.6 Hz, 1H, COCH2), 2.34 (t, J = 7.6 Hz, 2H,
COCH2), 3.57 (dd, J = 10.8, 5.2 Hz, 1H, H-1a), 3.62 (dd, J = 10.8,
7.2 Hz, 1H, H-1b), 3.65 (ddd, J = 10.0, 5.2, 2.4 Hz, 1H, H-50), 3.72
(dd, J = 11.6, 8.0 Hz, 1H, H-4a), 4.10 (ddd, J = 7.2, 6.0, 5.2 Hz, 1H,
H-2), 4.11 (dd, J = 11.6, 2.8 Hz, 1H, H-4b), 4.15 (dd, J = 12.4,
2.4 Hz, 1H, H-60a), 4.26 (dd, J = 12.4, 5.2 Hz, 1H, H-60b), 4.32 (ddd,
J = 8.0, 6.0, 2.8 Hz, 1H, H-3), 4.83 (d, J = 0.4 Hz, 1H, H-10), 5.07
(dd, J = 10.0, 3.2 Hz, 1H, H-30), 5.29 (dd, J = 10.0, 10.0 Hz, 1H, H-
40), 5.53 (dd, J = 3.2, 0.4 Hz, 1H, H-20). 13C NMR (100 MHz, CDCl3)
d: ꢀ5.48/ꢀ5.46 [Si(CH3)2], 13.8 (2C)/13.9/14.1 (acyl CH3), 18.2
[SiC(CH3)3], 22.2 (2C)/22.3/22.6 (acyl CH2), 24.3/24.4/24.5/25.0
(COCH2CH2), 25.3/27.9 [C(CH3)2], 25.8 [3C, SiC(CH3)3], 28.96/
29.00 [CO(CH2)2(CH2)2(CH2)2CH3], 31.2 (2C)/31.3/31.7 (acyl CH2),
33.95/33.98/34.02/34.2 (COCH2), 61.6 (C-1), 62.3 (C-60), 65.7 (C-
40), 68.4 (C-4), 68.7 (C-20), 71.1 (C-30), 72.6 (C-50), 76.8/77.2 (C-2
and C-3), 98.7 (C-10), 108.7 [C(CH3)2], 172.2/172.7/173.0/173.5
(OCOCH2). FABMS m/z (%): 881 ([M+Na]+, 8), 73 (100). FABHRMS
C
48H82O16Na [M+Na]+: 937.5501; found: 937.5511.
3.1.14. Preparation of acremomannolipin A homolog (1g)
To a solution of 20g (114 mg, 0.150 mmol) in THF (1.5 mL) was
added 1 M solution of TBAF in THF (0.23 mL, 0.23 mmol) at room
temperature, and the reaction mixture was stirred for 2 h. After
the reaction mixture was diluted with water, the resulting mixture
was extracted with EtOAc. The extract was washed with brine, and
concentrated in vacuo. The residue was purified by column chro-
matography (n-hexane–EtOAc, 3/1) to give 1g (98.1 mg, quant.) as
25
a colorless amorphous. [
a
]
ꢀ29.6 (c 1.05, CHCl3). IR (neat) cmꢀ1
:
D
3422, 1748, 1458, 1362, 1242, 1165, 1103, 1069. 1H NMR
(700 MHz, CDCl3) d: 0.882 (t, J = 7.2 Hz, 3H, acyl CH3), 0.885 (t,
J = 7.2 Hz, 6H, acyl CH3), 0.90 (t, J = 7.2 Hz, 3H, acyl CH3), 1.22–1.38
(m, 20H, acyl CH2), 1.51–1.59 (m, 2H, acyl CH2), 1.57/1.63 (each
ddt, J = 7.6, 7.6, 7.6 Hz, 2H, acyl CH2), 1.66 (tt, J = 7.6, 7.6, Hz, 2H, acyl
CH2), 2.20/2.21(each ddd, J = 15.6, 7.6 Hz, 1H, COCH2), 2.26/2.29/
2.34/2.36 (each dt, J = 15.6, 7.6 Hz, 1H, COCH2), 2.44 (t, J = 7.6 Hz,
2H, COCH2), 3.69 (ddd, J = 12.6, 5.8, 2.8 Hz, 1H, H-2a), 3.71 (ddd,
J = 10.0, 6.2, 2.8 Hz, 1H, H-50), 3.72 (ddd, J = 12.6, 5.8, 2.8 Hz, 1H, H-
2b), 3.83 (ddd, J = 11.2, 5.8, 2.8 Hz, 1H, H-1a), 3.87 (ddd, J = 11.2,
5.8, 2.8 Hz, 1H, H-1b), 4.19 (dd, J = 12.3, 6.2 Hz, 1H, H-60a), 4.22
(dd, J = 12.3, 2.8 Hz, 1H, H-60b), 4.70 (d, J = 1.0 Hz, 1H, H-10), 5.08
(dd, J = 10.0, 3.2 Hz, 1H, H-30), 5.26 (dd, J = 10.0, 10.0 Hz, 1H, H-40),
5.52 (dd, J = 3.2, 1.0 Hz, 1H, H-20).13C NMR (125 MHz, CDCl3) d:13.8
(2C)/13.9/14.0 (acyl CH3), 22.2 (2C)/22.3/22.6 (acyl CH2), 24.3/
24.4/24.5/25.0 (COCH2CH2), 28.9/29.0 [CO(CH2)2(CH2)2(CH2)2CH3],
31.1 (2C)/31.2/31.7 (acyl CH2), 33.88/33.91/34.0/34.1 (COCH2),
61.9 (C-2), 62.4 (C-60), 65.7 (C-40), 68.6 (C-20), 70.7 (C-30), 72.6 (C-
50), 73.3 (C-1), 99.4 (C-10), 172.3/172.6/173.2/173.4 (OCOCH2).
FABMS m/z (%): 667 ([M+Na]+, 98), 99 (100). FABHRMS calcd for
C34H60O11Na [M+Na]+: 667.4033; found: 667.4022.
calcd for C45H82O13SiNa [M+Na]+: 881.5422; found: 881.5446.
25
20j: A colorless oil. [
a
]
ꢀ29.6 (c 0.80, CHCl3). IR (neat) cmꢀ1
:
D
1748, 1458, 1373, 1246, 1219, 1161, 1099, 1069, 1045. 1H NMR
(400 MHz, CDCl3) d: 0.83–0.92 (m, 12H, acyl CH3), 1.20–1.40 (m,
20H, acyl CH2), 1.35/1.37 [each s, 3H, C(CH3)2], 1.46 [s, 6H,
C(CH3)2], 1.51–1.71 (m, 8H, acyl CH2), 2.17/2.22/2.23/2.29/2.31/
2.35/2.40/2.46 (each dt, J = 16.0, 7.2 Hz, 1H, COCH2), 3.56 (dd,
J = 10.0, 4.8 Hz, 1H, H-5a), 3.59 (dd, J = 8.0, 6.8 Hz, 1H, H-1a), 3.65
(ddd, J = 10.0, 5.6, 2.4 Hz, 1H, H-50), 3.92 (dd, J = 10.0, 8.0 Hz, 1H,
H-5b), 4.09 (dd, J = 7.2, 5.6 Hz, 1H, H-3), 4.11 (dd, J = 8.0, 7.2 Hz,
1H, H-1b), 4.16 (dd, J = 12.0, 2.4 Hz, 1H, H-60a), 4.18 (ddd, J = 8.0,
5.6, 4.8 Hz, 1H, H-4), 4.24 (dd, J = 12.0, 5.6 Hz, 1H, H-60b), 4.24
(ddd, J = 7.2, 7.2, 6.8 Hz, 1H, H-4), 4.70 (s, 1H, H-10), 5.04 (dd,
J = 10.0, 3.2 Hz, 1H, H-30), 5.26 (dd, J = 10.0, 10.0 Hz, 1H, H-40),
5.49 (d, J = 3.2 Hz, 1H, H-20). 13C NMR (100 MHz, CDCl3) d: 13.8
(2C)/13.9/14.1 (acyl CH3), 22.2 (2C)n22.3n22.6 (acyl CH2), 24.2/
24.4/24.5/24.98 (COCH2CH2), 24.99/25.4/26.6/27.8 [C(CH3)2],
28.97/29.02 [CO(CH2)2(CH2)2(CH2)2CH3], 31.2 (2C)/31.3/31.7 (acyl
CH2), 33.9/33.95/34.00/34.1 (COCH2), 62.3 (C-60), 65.6 (C-40), 66.4
(C-1), 67.7 (C-5), 68.4 (C-20), 71.0 (C-30), 72.7 (C-50), 74.1 (C-2),
74.9 (C-4), 78.5 (C-3), 98.5 (C-10), 109.2/109.5 [C(CH3)2], 172.2/
172.6/172.9/173.4 (OCOCH2). FABMS m/z (%): 837 ([M+Na]+, 9),
99 (100). FABHRMS calcd for C43H74O14Na [M+Na]+: 837.4976;
3.1.15. Preparation of acremomannolipin A homologs (1h–1k)
A solution of 20h (29.8 mg, 0.0417 mmol) in 90% aqueous tri-
fluoroacetic acid (TFA, 1.0 mL) was stirred at 0 °C for 10 min.
After removal of the solvent, the residue was co-evaporated five
times with MeOH in vacuo. The residue was purified by column
chromatography (CHCl3–MeOH, 100/1) to give
3,4,6-tri-O-hexanoyl-2-O-octanoyl-b- -mannopyranoside
20.4 mg, 72%) as a colorless amorphous. [
D
-glycerol-3-yl
(1h,
ꢀ29.6 (c 0.67,
D
25
a
]
D
CHCl3). IR (neat) cmꢀ1: 3476, 1748, 1682, 1458, 1362, 1246,
1165, 1103, 1069. 1H NMR (700 MHz, CD3OD) d: 0.901 (t,
J = 7.4 Hz, 3H, acyl CH3), 0.902 (t, J = 7.2 Hz, 3H, acyl CH3), 0.91 (t,
J = 7.0 Hz, 3H, acyl CH3), 0.93 (t, J = 7.2 Hz, 3H, acyl CH3), 1.24–
1.44 (m, 20H, acyl CH2), 1.52–1.60 (m, 4H, acyl CH2), 1.63–1.70
(m, 4H, acyl CH2), 2.19/2.21 (each dt, J = 15.4, 7.4 Hz, 1H, COCH2),
2.27/2.32 (each dt, J = 15.9, 7.4 Hz, 1H, COCH2), 2.34/2.37 (each
dt, J = 15.8, 7.4 Hz, 1H, COCH2), 2.40/2.46 (each dt, J = 15.4,
7.2 Hz, 1H, COCH2), 3.48 (dd, J = 11.4, 5.8 Hz, 1H, H-3a), 3.53 (dd,
J = 11.4, 5.4 Hz, 1H, H-3b), 3.57 (dd, J = 10.4, 6.6 Hz, 1H, H-1a),
3.75 (dddd, J = 6.6, 5.8, 5.4, 4.4 Hz, 1H, H-2), 3.83 (ddd, J = 10.0,
4.4, 2.2 Hz, 1H, H-50), 3.88 (dd, J = 10.4, 4.4 Hz, 1H, H-1b), 4.15
(dd, J = 12.2, 2.2 Hz, 1H, H-60a), 4.27 (dd, J = 12.2, 4.4 Hz, 1H, H-
60b), 4.90 (d, J = 1.0 Hz, 1H, H-10), 5.16 (dd, J = 10.0, 3.2 Hz, 1H, H-
found: 837.4995.
24
20k: A colorless oil. [
a]
ꢀ20.2 (c 1.01, CHCl3). IR (neat) cmꢀ1
:
D
1748, 1458, 1377, 1246, 1219, 1161, 1096, 1069. 1H NMR
(400 MHz, CDCl3) d: 0.86–0.94 (m, 12H, acyl CH3), 1.20–1.41 (m,
20H, acyl CH2), 1.36 [s, 6H, C(CH3)2], 1.38/1.44/1.47/1.54 [each s,
3H, C(CH3)2], 1.50–1.67 (m, 8H, acyl CH2), 2.17/2.21/2.23/2.28/
2.30/2.34/2.38/2.44 (each dt, J = 15.6, 7.6 Hz, 1H, COCH2), 3.59 (dd,
J = 10.4, 5.2 Hz, 1H, H-7a), 3.66 (ddd, J = 10.0, 5.6, 2.4 Hz, 1H, H-50),
3.74 (dd, J = 7.6, 7.6 Hz, 1H, H-1a), 3.99 (dd, J = 10.4, 7.6 Hz, 1H, H-
7b), 4.03 (dd, J = 7.6, 6.0 Hz, 1H, H-1b), 4,09 (dd, J = 6.4, 6.4 Hz, 1H,
H-3), 4.11 (ddd, J = 7.6, 6.4, 6.0 Hz, 1H, H-2), 4.16 (dd, J = 12.0,
2.4 Hz, 1H, H-60a), 4.23 (dd, J = 12.0, 5.6 Hz, 1H, H-60b), 4.296 (ddd,
J = 7.6, 5.6, 5.2 Hz, 1H, H-6), 4.303 (dd, J = 6.4, 6.4 Hz, 1H, H-4),

3772
N. Tsutsui et al. / Bioorg. Med. Chem. 23 (2015) 3761–3773
24
30), 5.29 (dd, J = 10.0, 10.0 Hz, 1H, H-40), 5.48 (dd, J = 3.2, 1.0 Hz, 1H,
H-20). 13C NMR (175 MHz, CD3OD) d: 14.2 (2C)/14.3/14.5 (acyl
CH3), 23.36/23.38/23.41/23.8 (acyl CH2), 25.5/25.60/25.62/26.4
(COCH2CH2), 30.2/30.3 [CO(CH2)2(CH2)2(CH2)2CH3], 32.3/32.36/
32.44/33.0 (acyl CH2), 34.86/34.92/32.96/35.2 (COCH2), 63.1
(C-60), 64.3 (C-1), 66.9 (C-40), 70.4 (C-20), 72.2 (C-2), 72.5 (C-3),
72.7 (C-30), 73.5 (C-50), 100.3 (C-10), 173.8/173.9/174.7/175.0
(OCOCH2). FABMS m/z (%): 697 ([M+Na]+, 100). FABHRMS calcd
for C35H62O12Na [M+Na]+: 697.4139; found: 697.4169.
1k: A colorless amorphous. [
a
]
D
ꢀ27.7 (c 1.07, CHCl3). IR (neat)
cmꢀ1: 3383, 1748, 1246, 1169, 1103, 1069. 1H NMR (800 MHz,
CD3OD) d: 0.90 (t, J = 7.3 Hz, 9H, acyl CH3), 0.92 (t, J = 7.3 Hz, 3H, acyl
CH3), 1.24–1.42 (m, 20H, acyl CH2), 1.52–1.59 (m, 4H, acyl CH2),
1.63–1.70 (m, 4H, acyl CH2), 2.19/2.21/2.27/2.31/2.34/2.37 (each
dt, J = 15.8, 7.3 Hz, 1H, COCH2), 2.40/2.47 (each dt, J = 15.3, 7.3 Hz,
1H, COCH2), 3.63 (dd, J = 9.2, 1.8 Hz, 1H, H-3), 3.64 (d, J = 6.2 Hz,
2H, H-1), 3.69 (dd, J = 10.5, 6.4 Hz, 1H, H-7a), 3.75 (dd, J = 8.5,
0.9 Hz, 1H, H-5), 3.80 (ddd, J = 8.5, 6.4, 2.3 Hz, 1H, H-6), 3.83 (ddd,
J = 10.1, 4.3, 2.3 Hz, 1H, H-50), 3.89 (dd, J = 9.2, 0.9 Hz, 1H, H-4),
3.92 (td, J = 6.2, 1.8 Hz, 1H, H-2), 4.141 (dd, J = 10.5, 2.3 Hz, 1H,
H-7b), 4.143 (dd, J = 12.4, 2.3 Hz, 1H, H-60a), 4.28 (dd, J = 12.4,
4.3 Hz, 1H, H-60b), 4.93 (d, J = 0.9 Hz, 1H, H-10), 5.16 (dd, J = 10.1,
3.2 Hz, 1H, H-30), 5.30 (dd, J = 10.1, 10.1 Hz, 1H, H-40), 5.51 (dd,
J = 3.2, 0.9 Hz, 1H, H-20). 13C NMR (200 MHz, CD3OD) d: 14.2 (2C)/
14.3/14.5 (acyl CH3), 23.37/23.39/23.42/23.8 (acyl CH2), 25.5/25.59/
25.63/26.4 (COCH2CH2), 30.2/30.3 [CO(CH2)2(CH2)2(CH2)2CH3],
32.3/32.4/32.5/33.0 (acyl CH2), 34.8/34.9/35.0/35.2 (COCH2), 63.0
(C-60), 65.0 (C-1), 66.8 (C-40), 70.2 (C-4), 70.5 (C-20), 71.0 (C-5),
71.4 (C-3), 71.9 (C-6), 72.0 (C-2), 72.7 (C-30), 73.5 (C-50), 73.8 (C-7),
100.6 (C-10), 173.75/173.85/174.8/175.0 (OCOCH2). FABMS m/z (%):
817 ([M+Na]+, 54), 99 (100). FABHRMS calcd for C39H70O16Na
[M+Na]+: 817.4562; found: 817.4579.
Following the method described above, 20i (48.4 mg,
0.055 mmol), 20j (331 mg, 0.406 mmol), and 20k (74.0 mg,
0.808 mmol) were converted to
anoyl-2-O-octanoyl-b- -mannopyranoside (1i, 29.6 mg, 75%),
-arabinitol-5-yl 3,4,6-tri-O-hexanoyl-2-O-octanoyl-b- -manno-
pyranoside (1j, 195 mg, 65%), and -glycero- -galactoheptitol-1-
-mannopyranoside (1k,
D-erythritol-4-yl 3,4,6-tri-O-hex-
D
D
D
D
D
yl 3,4,6-tri-O-hexanoyl-2-O-octanoyl-b-
D
62.0 mg, 97%), respectively.
25
1i: A colorless oil. [
a]
ꢀ26.4 (c 0.49, CHCl3). IR (neat) cmꢀ1
:
D
3445, 1748, 1458, 1416, 1362, 1246, 1169, 1107, 1069. 1H NMR
(700 MHz, CD3OD) d: 0.90 (t, J = 7.2 Hz, 6H, acyl CH3), 0.91 (t,
J = 6.8 Hz, 3H, acyl CH3), 0.92 (t, J = 7.0 Hz, 3H, acyl CH3), 1.23–
1.43 (m, 20H, acyl CH2), 1.51–1.59 (m, 4H, acyl CH2), 1.62–1.71
(m, 4H, acyl CH2), 2.19/2.21 (each dt, J = 15.4, 7.4 Hz, 1H, COCH2),
2.27/2.31 (each dt, J = 15.8, 7.2 Hz, 1H, COCH2), 2.34/2.37 (each
dt, J = 15.8, 7.6 Hz, 1H, COCH2), 2.40/2.46 (each dt, J = 15.2,
7.2 Hz, 1H, COCH2), 3.52 (ddd, J = 6.8, 6.2, 3.6 Hz, 1H, H-2), 3.57
(dd, J = 11.4, 6.2 Hz, 1H, H-1a), 3.64 (dd, J = 10.0, 6.8 Hz, 1H, H-
4a), 3.67 (ddd, J = 6.8, 6.8, 2.2 Hz, 1H, H-3), 3.70 (dd, J = 11.4,
3.6 Hz, 1H, H-1b), 3.82 (ddd, J = 10.1, 4.4, 2.2 Hz, 1H, H-50), 4.05
(dd, J = 10.0, 2.2 Hz, 1H, H-4b), 4.14 (dd, J = 12.4, 2.2 Hz, 1H, H-
60a), 4.27 (dd, J = 12.4, 4.4 Hz, 1H, H-60b), 4.90 (d, J = 0.8 Hz, 1H,
H-10), 5.16 (dd, J = 10.1, 3.4 Hz, 1H, H-30), 5.29 (dd, J = 10.1,
10.1 Hz, 1H, H-40), 5.50 (dd, J = 3.4, 0.8 Hz, 1H, H-20). 13C NMR
(175 MHz, CD3OD) d: 14.2 (2C)/14.3/14.5 (acyl CH3), 23.36/23.38/
23.41/23.8 (acyl CH2), 25.5/25.60/25.62/26.4 (COCH2CH2), 30.2/
30.3 [CO(CH2)2(CH2)2(CH2)2CH3], 32.34/32.36/32.5/33.0 (acyl
CH2), 34.8/34.9/35.0/35.2 (COCH2), 63.1 (C-60), 64.6 (C-1), 66.8
(C-40), 70.5 (C-20), 72.4 (C-3), 72.7 (C-30), 73.1 (C-4), 73.5 (C-50),
73.6 (C-2), 100.5 (C-10), 173.78/173.84/174.7/175.0 (OCOCH2).
FABMS m/z (%): 727 ([M+Na]+, 30), 73 (100). FABHRMS calcd for
3.2. Bioassay of acremomannolipin A homologs3
The Schizosaccharomyces pombe strains used in this study are
calcineurin deletion cells (h+ leu1 ura4-D18 ppb1::ura4+), the pre-
paration of which is described in a Supplementary data. In the pre-
sent study the cells were provided by Dr. M. Yanagida. The cells
were grown at 27 °C in the YPD (the complete medium yeast
extract-peptone-dextrose) and harvested in the early logarithmic
growth phase. The cells were resuspended in fresh YPD, and the
optical density was adjusted to 0.5 at 660 nm. The cells were
spread onto the YPD agar medium containing 0.1 M MgCl2 and
then put a filter paper disk (3 mm in diameter) in each section.
Next, 5
l
L of DMSO or the compound which was dissolved in
DMSO at a concentration of 100
lM was spotted onto each paper
and the cells were incubated at 27 °C for 3 days.
36H64O13Na [M+Na]+: 727.4245; found: 727.4277.
Acknowledgements
C
24
1j: A colorless amorphous. [
a
]
D
ꢀ26.6 (c 1.11, CHCl3). IR (neat)
cmꢀ1: 3395, 1748, 1458, 1366, 1242, 1165, 1099, 1069. 1H NMR
(700 MHz, CD3OD) d: 0.901 (t, J = 7.4 Hz, 3H, acyl CH3), 0.902 (t,
J = 7.4 Hz, 3H, acyl CH3), 0.91 (t, J = 7.0 Hz, 3H, acyl CH3), 0.93 (t,
J = 7.2 Hz, 3H, acyl CH3), 1.24–1.42 (m, 20H, acyl CH2), 1.52–1.60
(m, 4H, acyl CH2), 1.63–1.70 (m, 4H, acyl CH2), 2.19/2.21/2.27/
2.32/2.34/2.37 (each dt, J = 15.8, 7.4 Hz, 1H, COCH2), 2.40/2.47
(each dt, J = 15.5, 7.2 Hz, 1H, COCH2), 3.48 (dd, J = 8.4, 2.0 Hz, 1H,
H-3), 3.61 (d, J = 6.2 Hz, 2H, H-1), 3.69 (dd, J = 10.8, 6.4 Hz, 1H,
H-5a), 3.78 (ddd, J = 8.4, 6.4, 2.8 Hz, 1H, H-4), 3.83 (ddd, J = 10.0,
4.4, 2.2 Hz, 1H, H-50), 3.86 (td, J = 6.2, 2.0 Hz, 1H, H-2), 4.11 (dd,
J = 10.8, 2.8 Hz, 1H, H-5b), 4.15 (dd, J = 12.4, 2.2 Hz, 1H, H-60a),
4.28 (dd, J = 12.4, 4.4 Hz, 1H, H-60b), 4.92 (d, J = 1.0 Hz, 1H, H-10),
5.16 (dd, J = 10.0, 3.2 Hz, 1H, H-30), 5.30 (dd, J = 10.0, 10.0 Hz, 1H,
H-40), 5.51 (dd, J = 3.2, 1.0 Hz, 1H, H-20). 13C NMR (175 MHz,
CD3OD) d: 14.2 (2C)/14.3/14.5 (acyl CH3), 23.38 (2C)/23.42/23.8
(acyl CH2), 25.5/25.60/25.62/26.3 (COCH2CH2), 30.2/30.3
[CO(CH2)2(CH2)2(CH2)2CH3], 32.3/32.4/32.5/33.0 (acyl CH2), 34.8/
34.9/35.0/35.2 (COCH2), 63.1 (C-60), 64.8 (C-1), 66.8 (C-40), 70.5
(C-20), 71.67 (C-2), 71.69 (C-4), 72.1 (C-3), 72.7 (C-30), 73.48
(C-50), 73.50 (C-5), 100.6 (C-10), 173.77/173.84/174.8/175.0
(OCOCH2). FABMS m/z (%): 757 ([M+Na]+, 100). FAB HRMS calcd
for C37H66O14Na [M+Na]+: 757.4350; found: 757.4342.
This study was performed as a research program of the Project
for Development of Innovative Research on Cancer Therapeutics
(P-Direct), Ministry of Education, Culture, Sports, Science and
Technology of Japan (MEXT). This study was supported by research
grants from MEXT, and by the MEXT-Supported Program for the
Strategic Research Foundation at Private Universities (2014–2018).
We would like to thank Dr. M. Yanagida for providing the strain.
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2015.03.079.
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