Synthesis, conformational analysis and SAR research of OSW-1 analogues

Article abstract of DOI:10.1016/j.tet.2016.05.049

A series of novel OSW-1 analogues were synthesized by coupling disaccharides (2-O-4-methoxylbenzoyl-β-D-xylopyranosyl-(1→3)-2-O-acetyl-α-L-arabinopyranosyl) or (2-O-4-(E)-cinnamoyl-β-D-xylopyranosyl-(1→3)-2-O-acetyl-α-L-arabinopyranosyl) and their 1→4 lin

Full text of DOI:10.1016/j.tet.2016.05.049

Tetrahedron 72 (2016) 4091e4102
Contents lists available at ScienceDirect
Tetrahedron
journal homepage: www.elsevier.com/locate/tet
Synthesis, conformational analysis and SAR research of OSW-1
analogues
,
Chao Liu a, A-peng Wang a, Longlong Jin a, Yanshen Guo a, Yan Li b, Zhehui Zhao a
*
,
,
Pingsheng Lei a
*
a State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Beijing Key Laboratory of Active Substances Discovery and Drug
Ability Evaluation, Department of Medicinal Chemistry, Institute of Materia Medica, Peking Union Medical College, Chinese Academy of Medical
Sciences, Beijing 100050, PR China
b Department of Pharmacology, Institute of Material Medica, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100050,
PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
A
series of novel OSW-1 analogues were synthesized by coupling disaccharides (2-O-4-
Received 28 March 2016
Received in revised form 16 May 2016
Accepted 18 May 2016
methoxylbenzoyl-
b
-
D
-xylopyranosyl-(1/3)-2-O-acetyl-
-arabinopyranosyl) and their 1/4 linked analogues [(2-O-4-
-xylopyranosyl-(1/4)-2-O-acetyl- -arabinopyranosyl) or (2-O-4-(E)-cinnamoyl-
-arabinopyranosyl)] with three different steroidal sapogenins at
-hydroxy. Their conformation was analyzed with NMR spectroscopy and molecule simulation. The
a-L-arabinopyranosyl) or (2-O-4-(E)-cinnamoyl-
b
-D
-xylopyranosyl-(1/3)-2-O-acetyl-
a-
L
methoxylbenzoyl-
b
-D
a-L
Available online 20 May 2016
b-D-xylopyranosyl-(1/4)-2-O-acetyl-a-L
16
b
Keywords:
OSW-1 analogues
Synthesis
Conformation analysis
Molecule simulation
Antitumor activity
arabinose moiety of 1e3 linked analogues was in chair conformation and 1e4 linked analogues was in
boat conformation. 1e3 linked analogues exhibited potent anti-proliferation activity against a panel of
human tumor cells at nanomolar concentration level, while 1e4 linked analogues did not show anti-
tumor activity. This work should provide an evidence that the conformation plays an important role in
the antitumor activity.
Ó 2016 Elsevier Ltd. All rights reserved.
1. Introduction
such as mitomycin C, adriamycin, and taxol.2 The biological study
showed that OSW-1 can damage the membrane and cristae of
OSW-1 (1) and its related congener 2 (Fig. 1), isolated from the
bulbs of Ornithogalum saundersiae,1 have attracted considerable
attention because of their potent and selective antitumor activities
(IC50¼0.25 nM for OSW-1, IC50¼0.20 nM for congener 2, HL-60
cells) superior to those used in the clinical treatment of cancer
cancer cell’s mitochondria which played an important role in me-
diating the cell function, leading to the loss of transmembrane
potential, a significant increase of cytosolic calcium, and activation
of calcium-dependant apoptosis. The evidence disclosed that OSW-
1 may cause cancer cell’s death by apoptosis.3
A number of pharmacologic studies and synthetic of OSW-1
and its analogues have been reported.4e7 Kandutsch, A.8 and
Taylor9 discovered OSBP as a cytosolic receptor for oxysterols.
Hurley10 mentioned that sterol molecule binds within a hydro-
phobic tunnel in a manner consistent with a transport function for
ORPs (a family of OSBP). Burgett et al.11 identified that OSBP was
a high-affinity target of OSW-1 using affinity chromatography and
demonstrated that OSW-1 exerted its antitumor activity via OSBP.
Yan’s group also observed that ORP4L was a relevant target of
OSW-1 and played a key role in maintaining the proliferation ca-
pacity of certain malignant cell types.12 While Huang’s group
revealed that OSW-1 inhibited Naþ/Ca2þ exchanger 1, a membrane
protein, which led to apoptosis through increased calcium con-
centration in cytosol and mitochondria.13 The cellular targets and
the molecular mechanism of their action were still controversial.
Fig. 1. Antitumor glycosylsterols OSW-1 (1) and its congeners (2).
* Corresponding authors. Tel.: þ86 10 63162006; fax: þ86 10 63017757 (P.L.);
e-mail address: lei@imm.ac.cn (P. Lei).
http://dx.doi.org/10.1016/j.tet.2016.05.049
0040-4020/Ó 2016 Elsevier Ltd. All rights reserved.

4092
C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
The precise binding form of the receptor and OSW-1 remains
unclear. The relationship between the disaccharide chain and its
biological activity has not been well understood. For example, the
necessity of carbonyl group at C-22 was still controversial. Wojt-
kielewice et al. commented that the presence of a carbonyl group
at C-22 was pharmacophore required.7 However, B. Yu found that
the 22-deoxy-OSW-1 was slightly more potent than OSW-1.14 The
3D structures of OSW-1 (1) and its closely related congener (2)
were investigated by NMR studies and an X-ray crystallographic
analysis by Sakurai et al.15 They commented that the 3D structures
of OSW-1 (1) and (2) had a flat triangular molecular shape, which
was an important structural basis for its biological activity. In our
previous work16 an interesting result was found that 1e3 linked
analogues showed high activity, while 1e4 linked analogues
showed no activity. It was curious for us that the conformation of
inactive 1e4 linked analogues. We intend to affirm the relation-
ship between activity and conformation by comparing the con-
formation of 1e4 linked analogues to 1e3 linked analogues. Ten
1e3 linked (48a, 49a, 50a, 51a, 52a) and 1e4 linked (48b, 49b, 50b,
51b, 52b) OSW-1 analogues have been synthesized in our lab
(Fig. 2). Their conformation was carefully analyzed with NMR
spectroscopy and molecule simulation in this paper. This study
should facilitate the elucidation of SAR and be useful for designing
more effective analogues.
Scheme 1. Reagents and conditions: (a) TBDMSCl, DMAP, imidazole, DMF, 50 ꢀC,
30 min, 99%; (b) Oxone, NaHCO3, acetone/H2O/CH2Cl2, rt, 48 h, 94%; (c) Zn powder, KI,
AcOH, rt, 24 h, 64%; (d) TBDMSCl, DMAP, pyridine, rt, 6 h, 92%; (e) NaBH4/CeCl3$7H2O,
THF, rt, 14 h, 73%.
Scheme 2. Reagents and conditions: (a) Zn, HCl, ethanol, 50 ꢀC, 20 min, 71%; (b)
TBDMSCl, DMAP, pyridine, rt, 5 h, 90%.
2. Results and discussion
2.1. Chemistry
Three monosaccharides (two donors 13, 23 and one acceptor 29)
were designed and synthesised. The 4-methoxy benzoyl
substituted xylose 13 was synthesised following Yu’s route20,21
(Scheme 3).
Ten compounds were synthesized (48a, 48b, 49a,16 49b, 50a,
50b, 51a, 51b, 52a, 52b) by adapting the strategy of coupling sa-
pogenin with disaccharides (Fig. 2). Among them, cinnamoyl
substituted analogues (50a, 50b, 51a, 51b, 52a, 52b) and 22-OH-
OSW-1 analogues (48a, 48b) were reported for the first time.
Sapogenin 8 was synthesized in six steps by several groups
previously.16e18 The 3-OH of diosgenin was protected with tert-
butyldimethylsilyl group, and the C-16 and 5(6)-double bond were
oxy-functionalized with Oxone in the presence of NaHCO3 to afford
the mixture of epoxide diastereomers 3 and 4.18 In previous work,16
the C26eOH was protected with acetyl group during the F ring
opened in the acidic condition, and the acetyl group was removed
with CH3OH/CH3ONa to afford 5 and 6. We found that treating the
mixture of 3 and 4 with Zn/KI in AcOH, 5 and 6 could be directly
afforded by undergoing ring-chain tautomerization in acidic con-
dition (5:6¼5:1) (Scheme 1). The mixture could be separated by
flash column chromatography. 5 could be silylated to afford 7 in
a yield of 92%. Dione 7 was then reduced with NaBH4/CeCl3 to
Scheme 3. Reagents and conditions: (a) BnOH, HCl, 50 ꢀC, 24 h, 73%; (b) MBzCl, pyr-
idine, ꢁ15 ꢀC, 48 h, 65%; (c) TESCl, DMAP, pyridine, 93%; (d) H2, Pd/C, MeOH, 50 atm,
50 ꢀC, 24 h, 70%; (e) DBU, CCl3CN, CH2Cl2, ꢁ5 ꢀC, 5 h, 82%.
For synthesis of 2-cinnamoyl-
D
-xylose 23, monosaccharide
tetra-acetyl-
D
-xylose 14 was used as starting material22 (Scheme 4).
The thio ortho ester 16 was prepared via glycoside bromide 15.
After deacetylation and protection with 4-methoxybenzyl, 18 was
obtained. Thio ortho ester 18 was converted to thioglycoside 19 by
zinc chloride-promoted intramolecular ring opening in a yield of
87%. After deacetylation, cinnamoyl group was introduced at the C-
2 position to afford 21 in a yield of 61%, sulfide at anomeric position
was removed with NBS to afford 22, which was subsequently
converted to trichloroacetimidate 23 in a yield of 80%.23 This is the
provide 16b,22b-dihydroxyl aglycone 8. The absolute configuration
of C-22(S) was determined by Mosher’s method.16 Here using our
method, sapogenin 8 was produced in five steps with a better yield.
Sapogenin 10 was afforded from diosgenin via two steps.19
Compound 9 was prepared by reaction of Zn and hydrochloric
acid with diosgenin in ethanol under 50 ꢀC in a yield of 71%. Then
silylation of compound 9 with TBDMSCl in pyridine formed sapo-
genin 10 in a yield of 90% (Scheme 2).
first synthesis of 2-cinnamoyl-D-xylose trichloroacetimidate.
Scheme 4. Reagents and conditions: (a) 30% HBreAcOH, CH2Cl2, 0 ꢀC to rt, 4 h, 93%;
(b) EtSH, 2,6-lutidine, MeNO2, rt, 12 h, 82%; (c) CH3ONa, MeOH, 25 ꢀC, 3 h; (d) NaH,
DMF, PMBCl, reflux, 4 h, 90%; (e) ZnCl2, CH2Cl2, ꢁ60 ꢀC to 0 ꢀC, 87%; (f) CH3ONa, MeOH,
25 ꢀC, 4 h, 93%; (g) Cinnamic acid, DMAP, EDCI, CH2Cl2, rt, 24 h, 61%; (h) NBS, H2O,
CH2Cl2, rt, 1 h, 83%; (i) CCl3CN, DBU, CH2Cl2, ꢁ5 ꢀC, 5 h, 80%.
Fig. 2. Structures of synthesized OSW-1 analogues.

C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
4093
0
As an acceptor, the 1-thio-(4-methylphenyl)-2-O-acetyl-
a
-
L
L
-
-
the
d
H1 was about 4.3 ppm for C22-methylene or C22-hydrox-
0
arabinopyranoside 29 was readily prepared from tetra-acetyl-
ymethyl analogues (Table 2). This shift indicated that H1 of
arabinose for C22-carbonyl ana0logues located in shielding zone of
carbonyl and implied that H1 of arabinose was near by C22 of
side chain.
arabinose (24) in five steps as illustrated16 in Scheme 5.
Second, a significant difference between 48a and 48b was
0
0
that the coupling constant of H1 and H5 in the portion of
arabinose in 01H NMR. 48a arabinose moiety:
d (ppm): 4.27 (d,
J¼7.2 Hz, H1 ); 3.89 (dd, J¼3.0 Hz, J¼12.5 Hz, H5a0); 3.55 (dd,
Scheme 5. Reagents and conditions: (a) p-Thiocresol, BF3$Et2O, CH2Cl2, 0 ꢀC to rt, 8 h,
85%; (b) CH3ONa, CH3OH, rt, 2 h, 90%; (c) (CH3)2C(OCH3)2, CSA, DMF, under vacuum,
50 ꢀC, 5 h, 95%; (d) Ac2O, DMAP, Et3N, CH2Cl2, rt, 1.5 h, 90%; (e) Amberlite IR-120(Hþ),
CH3OH, 50 ꢀC, 4 h, 81%.
0
0
J¼1.3 Hz, J¼12.5 Hz, H5b ), while 48b: 4.32 (d, J¼5.5 Hz, H1 ); 4.00
0
(dd, J¼5.0 Hz, J¼12.0 Hz, H5a ); 3.53 (dd, J¼2.4 Hz, J¼12.0 Hz,
H
5b0) (Table 3). The data demonstrated that the arabinose part
was in chair conformation for 48a, but approximate in boat
conformation for 48b. In addition, this result was also confirmed
Glycosylation of diol acceptor 29 with xylosyl donor 13 or 23 in
presence of BF3$Et2O afforded corresponding disaccharides (30
and 31) or (32 and 33). Their structures were determined by 2D-
NMR. Then the compounds (30, 31, 32, 33) were exposed to sily-
lation using TESCl in pyridine, sulfides at anomeric position were
removed with NIS, disaccharides were then converted to tri-
chloroacetimidates (34, 35, 36, 37) under basic conditions
(Scheme 6).
0
by NOESY. A strong H10eH4 dipolar interaction could be ob-
served from 48b while is not expected in 48a (Supplementary
data). This phenomenon could be ascribed to a large sub-
stituent group attached at 4-position of arabinose part as
pseudoequatorial in a boat conformation for 48b. The skew boat
conformation was benefit for lower molecular energy and more
stable. The difference could also be observed in 50a and 50b, 51a
and 51b correspondingly.
With the promotion of trimethylsilyl triflate (TMSOTf), coupling
of 16
chloroacetimidates (34, 35, 36, 37) gave the corresponding 16-
glycosidation saponins (38, 39, 40, 41). Coupling of 16 -OH agly-
b
,22(S)-diol aglycone
8
with four disaccharide tri-
Third, compared the 13C spectra of 1e3 and 1e4 linked sapo-
genin analogues with their relative disaccharides, a significant
difference could be observed. dC3 of arabinose of 1e3 linked sa-
b
cone 10 with two disaccharide trichloroacetimidates (35, 37) gave
the corresponding 16-glycosidation saponins (42, 43). Then re-
moval of all protecting groups in two steps afforded target com-
pounds 48a, 48b, 50a, 50b, 52a, 52b (Scheme 7). The saponins 38,
39, 40, 41 were exposed to oxidation with pyridinium dichromate
(PDC) to afford 22-ones (44, 45, 46, 47). Removal of all protecting
groups in two steps afforded four target compounds 49a, 49b, 51a,
51b (Scheme 8) benefit for forming such hydrophilic portion and
hydrophobic cluster.
pogenins (48a, 50a, 51a) shifted to low field. The Dd was approxi-
mate 3.7e7.0 ppm. However,1e4 linked sapogenins (48b, 50b, 51b)
d
C4 shifted to high field (Table 4). The Dd was approximate
ꢁ4.0 ppm. 13C chemical shift is sensitive to stereochemistry. Space
effect is an important factor to the 13C chemical shift. This differ-
ence could be attributed to the conformation discrepancy of these
compounds.
Fourth, in the 2D-NOESY spectra for compounds 48e52, weak
NOE crosspeaks were observed between H27 and H33/35, H32/36, and
between H29 and H32/36 in compound 48a (Fig. 3). It indicated the
close proximity between the C200-p-methoxybenzoyl (p-MBz) group
2.2. Conformation analysis
0
and the C2 -acetyl group, the C27 methyl groups of the C20eC27
sterol side chain. Similarly for compound 51a, NOEs were detected
In order to study the conformational difference between 1e3
and 1e4 linked analogues, many assays had been made to cultivate
crystal. Unfortunately, we did not get the single crystal because of
their poor physicochemical properties. Structures of target com-
pounds were studied with 1H NMR, 13C NMR, DEPT, NOESY, COSY,
HSQC, HMBC. 13C and 1H signals for all compounds were assigned in
detail. Four significant differences were observed by analyzing the
NMR dates which showed the conformation was different between
between H27 and H31, H32, H35/37, H34/38 and between H29 and H31
,
H
35/37, H34/38. This phenomenon was consistent with Sakurai’s re-
search report,15 they suggested that the overall molecular shape of
OSW-1 was a flat right triangle. However, such correlation was not
found in 48b, neither in 49b, 50b, 51b and 52b. This meant the
conformation was different between the 1e3 linked and 1e4 linked
analogues.
The results of molecule simulation showed that (Fig. 4): the
conformation of arabinose in 1e3 linked analogues was the chair
conformation, while that of 1e4 linked analogues was the skew
boat conformation. These results were consistent with the
analysis of 1H NMR previously. Another phenomenon was ob-
served in the molecule simulation, acetyl and side chain C27
the 1e3 linked and 1e4 linked analogues.
The first significant difference was that
analogues (49a, 49b, 51a, 51b) shifted to high-field compared
0
d
H1 of C22-carbonyl
with C22-methylene (52a, 52b) or C22-hydroxymethyl (48a, 48b,
0
50a, 50b) analogues. The
d
H1 of arabinose moieties was
about 4.0 ppm for C22-carbonyl analogues (Table 1), while
Scheme 6. Reagents and conditions: (a) 4 A MS, BF3$Et2O, CH2Cl2, ꢁ60 ꢀC, 3 h, 36% for 30, 18% for 31, 21% for 32, 13% for 33; (b) TESCl, DMAP, pyridine; (c) NIS, H2O, CH2Cl2, 1 h; (d)
CCl3CN, DBU, CH2Cl2, 3 h, three steps 56% for 34, 61% for 35, 64% for 35, 52% for 37.
ꢀ

4094
C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
Scheme 7. Reagents and conditions: (a) 4 A MS, TMSOTf, CH2Cl2, ꢁ60 ꢀC, 3 h, 64% for 38, 58% for 39, 51% for 40, 47% for 41, 55% for 42, 43% for 43; (b) i) DDQ, Pd(CH3CN)2Cl2, CH2Cl2/
actone/H2O, rt, 5 h, 59% for 50a, 54% for 50b, 48% for 52a, 61% for 52b; ii) Pd(CH3CN)2Cl2, actone/H2O, rt, 3 h, 59% for 48a, 67% for 48b.
ꢀ
Scheme 8. Reagents and conditions: (a) PDC, CH2Cl2, rt, 6 h, 96% for 44, 67% for 45, 80% for 46, 74% for 47; (b) i) DDQ, Pd(CH3CN)2Cl2, CH2Cl2/actone/H2O, rt, 5 h, 52% for 51a, 53% for
51b; ii) Pd(CH3CN)2Cl2, actone/H2O, rt, 3 h, 62% for 49a, 57% for 49b.
Table 1
functionalities. On the contrary, the results of molecule simula-
tion showed that the distance of acetyl and the side chain and
0
dH1 of analogues which C22 is carbonyl
aromatic ring was farther in 1e4 linked analogues. The distance
49a
49b
3.96
51a
51b
ꢀ
ꢀ
0
of aromatic ring to acetyl was 5.59 A, that to H27 was 3.88 A, and
the arabinose was the skew boat conformation. The change of
arabinose conformation led to the overall molecular conforma-
tion changed, which had a non flat triangular structure com-
pared with 1e3 linked analogues. Those data demonstrated that
the 1e4 linked analogues presented disperse structure which
was not benefit for forming such hydrophilic portion and hy-
drophobic cluster.
d
H1
4.00
4.06
4.09
Table 2
0
H1 of analogues which C22 is methylene or hydroxymethyl
d
48a
48b
50a
50b
52a
52b
0
d
H1
4.29
4.31
4.27
4.32
4.28
4.27
Table 3
The 1H NMR analysis of arabinose fragment for analogues (48a, 48b, 50a, 50b, 51a,
51b)
methyl and xylose substituted aromatic ring were close to each
other in 1e3 linked analogues, the distance of aromatic ring to
48a
d
48b d
0
0
H1
H5
4.27 (d, J¼7.2)
4.32 (d, J¼5.5)
ꢀ
ꢀ
acetyl was 3.11 A, that to H27 was 3.35 A. These results were
conformity with the NOESY research previously. The 1H NMR
3.89 (dd, J¼3.0, J¼12.5)
3.55 (dd, J¼1.3, J¼12.5)
4.00 (dd, J¼5.0, J¼12.0)
3.53 (dd, J¼2.4, J¼12.0)
0
spectra showed that C22 was close to H1 . Based on the results of
50a
d
50b d
1H NMR and molecule simulation, we speculated that the 1e3
linked analogues had flat triangular structures. The flat tri-
angular conformation of 1e3 linked 0a0 nalogues had a hydrophilic
0
0
H1
H5
4.29 (d, J¼7.1)
4.31 (d, J¼6.0)
3.87 (dd, J¼3.0, J¼12.6)
3.99 (dd, J¼4.5, J¼12.1)
3.54 (dd, J¼1.3, J¼12.6)
3.54 (dd, J¼2.2, J¼12.1)
portion which composed by O300, O4 and O5 of the xylose res-
00
51a
d
51b d
idue and O4 of arabinose residue. The C200-acylate and C2 -ace-
tate groups and the C20eC27 sterol side chain pack against each
other which resulted the generation of a hydrophobic cluster.
This overall conformation was important for its biological
0
0
0
0
H1
H5
4.06 (d, J¼6.9)
4.09 (d, J¼5.7)
3.85 (dd, J¼3.1, J¼12.5)
3.49 (dd, J¼1.6, J¼12.5)
3.94 (dd, J¼4.6, J¼11.9)
3.49 (dd, J¼2.0, J¼11.9)

C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
4095
and 7.2ꢂ10ꢁ11 mol/L to Pfeiffer, respectively. Analogues 52a and
52b were tested against HCT-116, NCI-H1975, Capan2, SW1990, SK-
N-SH, BGC-823, and HepG2 cell lines (Table 7). Activity of 52a was
also much higher than 52b, the IC50 value of 52a was
1.2ꢂ10ꢁ12 mol/L to HepG2, 2.1ꢂ10ꢁ12 mol/L to BGC823, while 52b
showed inactivity to all of those tumor cells.
An interesting phenomenon was observed from antitumor ac-
tivity results, the higher antiproliferative activity was displayed by
compounds containing 1e3 linked disaccharide which had flat
triangular structures (49a, 51a, 52a) at nanomolar concentration
level.1e4 linked analogues which had non flat triangular structures
(48b, 49b, 51b, 52b) showed a lower activity. The key role of flat
triangular molecular shape of 1e3 linked OSW-1 analogues for the
high activity was confirmed. Notably, the activity of 52a which
aglycone C22 was methylene was higher compared with 49a and
51a which C22 was carbonyl. This indicated that the C22 of choles-
tane side chain which reduction form seemed could boost its ac-
tivity by almost 10 times.
Table 4
The 13C NMR analysis of arabinose fragment
Compound
Position
Unlinkeda
Linkedb
Ddc
7.0
0
0
0
0
0
0
48a
48b
50a
50b
51a
51b
C3
C4
C3
C4
C3
C4
74.0
75.0
75.9
75.3
75.9
75.3
81.0
71.0
79.6
71.1
80.8
71.2
ꢁ4.0
3.7
ꢁ4.2
4.9
ꢁ4.1
a
Disaccharide which did not attach to saponin.
b
c
Target compound which disaccharide attached to saponin.
Dd dlinked dunlinked
¼
ꢁ
.
3. Conclusion
In conclusion, 10 OSW-1 analogues were synthesized and the
conformation of analogues was studied by NMR analysis and
molecule simulation. The arabinose moiety of 1e3 linked analogues
was in chair conformation and 1e4 linked analogues was in boat
conformation. The antitumor activity of 1e3 linked analogues
which present flat triangular structure was much higher than that
of 1e4 linked analogues which present non flat triangular struc-
ture. From the relationship between conformation and activity,
a conclusion was presumed that the flat triangular conformation of
OSW-1 analogues played a key role in their high antitumor activity.
The non flat triangular conformation which showed by 1e4 linked
analogues was unfavorable for its activity.
Fig. 3. Selected NOESY correlations (600 MHz, CD3OD) for compounds 48a (a) and 51a
(b). NOEs shown by black and red arrows (the corresponding crosspeaks in the NOESY
spectra are shown in bottom panels).
2.3. Antitumor activities in vitro
The synthetic OSW-1 analogues 49a, 49b, 51a, 51b, were
screened against HCT-116, MDA-MB-231, A375, BGC-823, ACHN,
SW1990, A2780 and HepG2 cell lines by MTT assay. Among them
49a and 49b were considered as positive control. Both 49a and 51a
were far more effective against all of the tested tumor cells than
49b and 51b (Table 5) in which of BGC823 and A2780 were most
sensitive at nanomolar concentration level. The activity test of 49a
and 51a were repeated three times to insure their accuracy. The
cytotoxicity (IC50) value of 49a was 2.7ꢂ10ꢁ10 mol/L to BGC823 and
1.2ꢂ10ꢁ10 mol/L to A2780, the value of 51a was 6.8ꢂ10ꢁ10 mol/L to
BGC823 and 7.1ꢂ10ꢁ10 mol/L to A2780. While the value of 49b was
only 2.7ꢂ10ꢁ5 mol/L to BGC823 and 1.9ꢂ10ꢁ6 mol/L to A2780, the
value of 51b was 3.7ꢂ10ꢁ5 mol/L to BGC823 and 2.2ꢂ10ꢁ5 mol/L to
A2780. In order to verify the reliability and expand the types of
tumor cell lines, another test was repeated, which showed high
anti-proliferation activities for 49a and 51a to SH-SY5Y, SK-N-SH,
T98G, Daoy, NCI-H1975, NCI-H1703, HCT116, BGC823, Capan2,
SW1990, HepG2, A375, ACHN, MDA-MB-231, A2780, PC-3 and
Pfeiffer cell lines (Table 6). Pfeiffer was the most sensitive cell line to
49a and 51a, the IC50 values of 49a and 51a were 5.3ꢂ10ꢁ9 mol/L
This study presents a considerable opportunity for the de-
velopment of new OSW-1 analogues as antitumor candidates.
4. Experimental section
4.1. Materials and method in biological tests
4.1.1. Materials. Human cancer cell lines, SH-SY5Y, SK-N-SH, T98G,
Daoy, NCI-H1975, NCI-H1703, HCT116, BGC823, Capan2, SW1990,
HepG2, A375, ACHN, MDA-MB-231, A2780, PC-3, and Pfeiffer were
purchased from the American Type Culture Collection (ATCC) and
Cell Culture Center of Institute of Basic Medical Sciences, Chinese
Academy of Medical Sciences. All the cell lines were cultured in
RPMI-1640 media supplemented with 10% fetal calf serum, peni-
cillin (100
mg/mL) and streptomycin (100 mg/mL) (Gibco BRL, NY,
USA), and incubated at 37 ꢀC in a humidified air atmosphere
Fig. 4. Molecule simulation schematic for 49a (a) and 49b (b).

4096
C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
Table 5
Antitumor activities of compounds 49a, 49b, 51a, 51b against tumor cellsa
OSW-1 derivative
Cell line; IC50 (mmol/L)
HCT116 MDA-MB-231
0.0086 0.015
A375
BGC823
ACHN
0.03
28
0.0026
>50
SW1990
A2780
HepG2
49a
49b
51a
51b
0.038
18
0.045
20
0.00027
27
0.00068
37
0.71
>50
0.026
>50
0.00012
1.9
0.00071
22
0.15
23
0.16
>50
24 24
0.0053
>50
0.00012
2.60
a
The antitumor activities against HCT-116 (colon carcinoma), MDA-MB-231 (breast cancer), A375 (melanoma cell), BGC-823 (stomach carcinoma), ACHN (renal cancer),
SW1990 (pancreatic cancer), A2780 (ovarian cancer) and HepG2 (liver cancer) cell lines in vitro were evaluated by the MTT assay.
Table 6
Antitumor activities of compounds 49a and 51a against tumor cellsa
IC50(mmol/L)
SH-SY5Y
SK-N-SH
T98G
Daoy
NCI-H1975
NCI-H1703
HCT116
BGC823
Capan2
49a
51a
0.025
0.0089
0.63
0.48
0.0019
0.0008
0.0035
0.00023
0.075
0.002
0.021
0.0092
0.024
0.073
0.00027
0.00068
1.9
0.43
SW1990
HepG2
A375
ACHN
MDA-MB-231
A2780
PC-3
Pfeiffer
d
49a
51a
0.71
0.027
0.14
0.16
0.038
0.045
0.03
0.0026
0.015
0.00012
0.00018
0.00073
0.13
0.095
0.0053
0.000072
d
d
a
The antitumor activities against SH-SY5Y, SK-N-SH (neuroblastoma), T98G (glioblastoma), Daoy (medulloblastoma), NCI-H1975, NCI-H1703 (lung cancer), HCT116 (colon
cancer), BGC823 (gastric cancer), Capan2, SW1990 (pancreatic cancer), HepG2 (HCC) A375 (melanoma), ACHN (RCC), MDA-MB-231 (breast cancer), A2780 (ovarian cancer),
PC-3 (prostate cancer), and Pfeiffer (lymphoma) cell lines in vitro were evaluated by the MTT assay.
Table 7
Antitumor activities of compounds 52a and 52b against tumor cellsa
OSW-1 derivative
Cell line; IC50 (mmol/L)
HCT116
NCI-H1975
0.00012
>1
Capan2
SW1990
SK-N-SH
BGC823
HepG2
52a
52b
0.00043
>1
0.024
>1
0.013
>1
0.0012
>1
0.000021
>1
0.0000012
>1
a
The antitumor activities against HCT-116 (colon carcinoma), NCI-H1975 (lung cancer), Capan2 (pancreatic cancer), SW1990 (pancreatic cancer), SK-N-SH (neuroblastoma),
BGC-823 (stomach carcinoma), and HepG2 (liver cancer) cell lines in vitro were evaluated by the MTT assay.
containing 5% CO2. All cells were harvested in their exponentially
growing phase.
170e180 ꢀC for 10e15 h before use. Commercial anhydrous solvents
and reagents were used without further purification.
4.1.2. MTT assay. Cells were seeded into 96-well plates at a con-
centration of 1000e2000 cells/well and left there to adhere the
plate overnight. Then cells were incubated for another 72 h in the
absence and presence of various compounds and each condition
was performed with three wells. After removal of supernatant, cells
were treated with 0.5 mg/mL MTT for 4 h, the purple blue sediment
4.3. Method of molecule simulation
The partial initial conformation of the molecule comes from the
published crystal structure of 215 (Fig. 1), the initial conformation of
1e3 linked analogues was constructed based on the three-
dimensional crystal structures of 2, 1e4 linked analogues was
manually built from atoms using the Sketch module of Sybyl (Tri-
pos Inc., St. Louis, MO, USA) on Dell workstation. They were mini-
mized by the Maximin module of Sybyl using Tripos force field and
partial charges of GasteigereMarsili method implemented in
Sybyl.24 The conformational searches were also applied before the
minimization to get the possible different low energy
conformation.
was dissolved in 200 mL DMSO. The relative optical density (OD)/
well was determined at a test wavelength of 570 nm in a WELL-
SCAN MK3 ELIASA (Labsystems, Dragon, Finland) using a 450 nm
reference filter. The 50% inhibitory concentration (IC50) was calcu-
lated from the linear equation, which was educed by concentration
versus growth inhibition regression curve.
4.2. Synthetic experiments and spectra data
4.4. Chemical synthesis
General Methods: Optical rotatings were recorded in a Per-
kineElmer 341 LC polarimeter. 1H NMR spectra were recorded at
4.4.1. Purity. Purity of final compounds 48a, 48b, 49a, 49b, 51a,
51b, 52a, 52b were found to be ꢃ95% as determined by HPLC
analysis. HPLC was performed with LabAlliance apparatus com-
prising pumps (III Pump series), a UVevis detector (525 dual
wavelength), and injection valve (Rheodyne model 7725i). Ana-
lytical HPLC was carried out with a Phenomenex Luna C18 column
300, 400, 500 or 600 MHz with chemical shifts reported in ppm (
in reference to the solvent peak. 13C NMR spectra were recorded at
75, 100, 125 or 150 MHz with chemical shifts reported in ppm ( ) in
d)
d
reference to the solvent peak. The coupling constants (J) are re-
ported in hertz (Hz). HRMS were recorded on an Agilent 1100 series
LC/MSD TOF. Thin layer chromatography (TLC) was performed on
silica gel GF254 plates detected by charring with 5% sulfuric acid in
ethanol solution. Chromatography was performed on silica gel (HG/
T2354-92) or 200e300 mesh and Sephadex LH-20 (GE Healthcare).
(5 mm), 0.46 cme25 cm. The eluting solvent was MeCN/water. The
flow rate was 0.8 mL/min. The elution pattern of products was
monitored continuously by UV at 272 nm. The purity and retention
times of final products were as follows: 48a (97.4%, 11.53 min,
MeCN/water 60:40), 48b (95.6%, 12.91 min, MeCN/water 60:40),
ꢀ
Molecular sieves 4 A used were dried under high vacuum at

C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
4097
49a (95.2%, 20.78 min, MeCN/water 45:55), 49b (97.1%, 27.39 min,
MeCN/water 40:60), 51a (98.6%, 11.00 min, MeCN/water 60:40),
51b (95.8%, 20.17 min, MeCN/water 45:55), 52a (99.3%, 8.78 min,
MeCN/water 65:35), 52b (98.1%, 21.95 min, MeCN/water 45:55).
H-1), 4.06 (dd, J¼4.1 Hz, J¼11.6 Hz, 1H, H-5a), 3.79 (s, 3H), 3.67 (s,
3H), 3.65e3.63 (m, 2H), 3.29 (dd, J¼9.3 Hz, J¼11.6 Hz, 1H, H-5b),
2.67 (m, 2H), 1.23 (t, J¼7.5 Hz, 3H). 13C NMR (100 MHz, CDCl3):
d
165.5, 159.3, 159.0, 145.4, 134.2, 130.3, 130.2, 130.0, 129.6, 129.4,
128.8, 128.1, 128.1, 117.5, 113.8, 113.6, 83.8, 80.9, 76.9, 74.1, 72.7, 71.1,
66.9, 55.1, 55.0, 24.0, 14.7. C32H36O7S, HRMS (ESI): calcd for
(MþHþ): 565.2260; found: 565.2275.
4.4.2. 3b-O-tert-Butyldimethylsilyl kryptogenin (5)/3b-O-tert-bu-
tyldimethylsilyl 16-hydroxyl-diosgenin (6). A solution of 3 and 4
(0.50 g, 0.89 mmol) in 10 mL HOAc was added KI (0.72 g, 8.9 mmol),
Zn (0.55 g, 4.45 mmol), after stirring for 20 h at room temperature
and then filtered. The filtrates were diluted with 30 mL CH2Cl2, the
solution was washed with water, saturated NaHCO3, brine, dried
over MgSO4, the concentrate was purified by column chromatog-
raphy on silica gel (petroleum ether: ethyl acetate¼5:1) to afford
compound 5 and 6 (0.29 g, 60% for 5, 0.058 g, 12% for 6).
4.4.8. 3,4-Di-O-(4-methoxybenzyl)-2-O-[(E)-cinnamoyl]-D-xylopyr-
anose (22). NBS (0.24 g, 1.10 mmol) was added in a solution of 21
(0.50 g, 0.88 mmol) in 10 mL CH2Cl2 with 0.50 mL H2O, after stirring
1 h at room temperature, then saturated Na2SO3 aq was added in
and stirred 15 min the solution was washed with NaHCO3, brine,
dried over MgSO4, the concentrate was purified by column chro-
matography on silica gel (petroleum ether:ethyl acetate¼5:1) to
4.4.3. 3
b
,26-O-Bis(tert-butyldimethylsilyl) kryptogenin (7). To a so-
afford compound 22 (0.38 g, 83%). 1H NMR (300 MHz, CDCl3):
d 7.75
lution of 5 (1.00 g, 1.80 mmol), DMAP (0.24 g, 1.80 mmol) in dry
pyridine (10 mL) was added TBDMSCl (0.55 g, 3.60 mmol). After
being stirred for 6 h, then saturated Na2SO3 aq was added in and
stirred 5 min. The solution was diluted with 50 mL CH2Cl2 and
washed with NaHCO3, brine, then dried over MgSO4 and concen-
trated in vacuo, the residue was purified by column chromatogra-
phy on silica gel (petroleum ether:ethyl acetate¼8:1) to afford
compound 7 (1.10 g, 92%).
(d, J¼16.0 Hz, 1H), 7.52 (m, 2H), 7.40 (m, 3H), 7.30e7.24 (m, 4H),
6.90 (d, J¼8.6 Hz, 2H), 6.87 (d, J¼8.6 Hz, 2H), 6.49 (d, J¼16.0 Hz, 1H),
6.20 (d, J¼2.5 Hz, 1H, H-1), 4.77 (dd, AB, J¼11.0, J¼40.7 Hz, 2H), 4.62
(dd, AB, J¼11.2, J¼17.8 Hz, 2H), 4.30 (t, J¼6.7 Hz, 1H, H-2), 3.86 (m,
1H), 3.80 (s, 3H), 3.80 (s, 3H), 3.73 (m, 1H), 3.62 (m, 1H), 2.60 (d,
J¼5.2 Hz, 1H, OH). 13C NMR (75 MHz, CDCl3):
d 165.4, 159.4, 159.3,
146.3, 134.0, 130.8, 130.5, 130.2, 129.6, 129.5, 128.9, 128.8, 128.2,
117.1, 113.9, 91.9, 79.4, 76.2, 74.3, 72.5, 70.3, 65.5, 62.8, 55.2.
C
30H32O8, HRMS (ESI): calcd for (MþHþ): 521.2175; found:
4.4.4. (22S,25R)-3
b
,26-Bis (tert-butyldimethylsilyloxy)-16,22-
521.2181.
dihydroxyl-cholest-5-ene (8). A solution of 7 (2.70 g, 4.00 mmol)
in 15 mL dry THF was added NaBH4 (7.50 g, 0.20 mol), CeCl3$7H2O
(5.90 g, 0.016 mol), after stirring for 14 h at room temperature the
solution was diluted with 100 mL CH2Cl2, the solution was washed
with saturated NaHCO3, brine, dried over MgSO4, the concentrate
was purified by column chromatography on silica gel (petroleum
ether:ethyl acetate¼3:1) to afford compound 8 (2.00 g, 73%).
4.4.9. 3,4-Di-O-(4-methoxybenzyl)-2-O-[(E)-cinnamoyl]-a/b-D-xylo-
pyranosyl trichloroacetimidate (23). The compound 22 (0.93 g,
1.80 mmol) was dissolved in dried CH2Cl2, added CNCCl3 (1.80 mL,
0.018 mol) and DBU (0.067 mL, 0.52 mmol) under Ar at -5 ꢀC, after
stirring for 5 h, the concentrate was purified by column chroma-
tography on silica gel (petroleum ether:ethyl acetate¼10:1) to af-
ford compound 23 (1.10 g, 80%). 1H NMR (400 MHz, CDCl3):
d 8.53
4.4.5. (25R)-Cholest-5-ene-3
b
,16
b
,26-triol (9). A solution of dio-
(s, 1H, NH), 7.60 (d, J¼16.0 Hz, 1H), 7.48 (m, 2H), 7.39 (m, 3H), 7.27
(d, J¼8.8 Hz, 2H), 7.22 (d, J¼8.8 Hz, 2H), 6.88 (d, J¼8.6 Hz, 2H), 6.78
(d, J¼8.6 Hz, 2H), 6.42 (d, J¼3.6 Hz, 1H, H-1), 6.28 (d, J¼16.0 Hz, 1H),
5.10 (dd, J¼3.6, J¼9.8 Hz, 1H), 4.82 (d, AB, J¼11.1 Hz, 1H), 4.72 (dd,
AB, J¼3.9, J¼11.1 Hz, 2H), 4.61 (d, AB, J¼11.1 Hz, 1H), 4.30 (t,
J¼6.7 Hz, 1H), 4.06 (t, J¼8.4 Hz, 1H), 3.83e3.74 (m, 2H), 3.80 (s, 3H),
sgenin (2.00 g, 4.80 mmol) in 300 mL dry EtOH, Zn powder (45.00 g,
0.69 mol) was added under drastic stirring, heating to 50 ꢀC, then
dropping dilute hydrochloric acid to the solution during 40 min,
after stirred for 30 min, and then filtered. The filtrates washed with
water, saturated NaHCO3, brine, dried over MgSO4, the concentrate
was purified by column chromatography on silica gel (petroleum
ether:ethyl acetate¼4:1) to afford compound 9 (1.40 g, 71%).
3.68 (s, 3H).
C
32H32Cl3NO8, HRMS (ESI): calcd for (MþHþ):
664.1272; found: 664.1271.
4.4.6. (25R)-3b,26-Bis
(tert-butyldimethylsilyloxy)-cholest-5-en-
4.4.10. 4-Methylphenyl (2-O-(4-methoxybenzoyl) -3,4-di-(O-trie-
16 -ol (10). To a solution of 9 (7.80 g, 0.019 mol) in dry Pyridine
b
thylsilyl)-
arabinopyranoside(30)/4-methylphenyl (2-O-(4-methoxybenzoyl)-
3,4-di-(O-triethylsilyl)- -xylopyranosyl-(1/3)-2-O-acetyl-4-hy-
droxy-1- -arabinopyranoside (31). The mixture of compound 13
b-D-xylopyranosyl-(1/4)-2-O-acetyl-3-hydroxy-1-a-L-
(80 mL) was added imidazole (5.40 g, 0.079 mol), DMAP (0.23 g,
0.19 mmol) and TBDMSCl (8.40 g, 0.056 mol). After being stirred for
5 h at room temperature, then saturated NaHCO3 aq was added in
and stirred 5 min. The solution was diluted with 500 mL CH2Cl2 and
washed with 1 mol/L HCl, saturated NaHCO3, brine, then dried over
NaSO4 and concentrated in vacuo, the residue was purified by
column chromatography on silica gel (petroleum ether:ethyl
acetate¼8:1) to afford compound 10 (11 g, 90%).
b-D
a-L
ꢀ
(0.58 g, 0.88 mmol) and 29 (0.40 g, 1.30 mmol) was added 4 A MS in
dried CH2Cl2 under Ar at ꢁ60 ꢀC, then the BF3$Et2O (0.011 mL,
0.044 mmol) was slowly added, after stirring for 3 h, Et3N was
added, MS was filtered and filtrate was concentrated, then the
residue was purified by column chromatography on silica gel (pe-
troleum ether:ethyl acetate¼8:1) to afford compound 30 (0.25 g,
4.4.7. Ethyl 3,4-di-O-(4-methoxybenzyl)-2-O-[(E)-cinnamoyl]-1-
36%) and 31 (0.13 g, 18%). (30) 1H NMR (300 MHz, CDCl3):
d 8.02 (d,
thio-
b
-
D
-xylopyranoside (21). A solution of 20 (0.20 g, 0.46 mmol)
J¼9.0 Hz, 2H), 7.36 (d, J¼8.0 Hz, 2H), 7.08 (d, J¼8.0 Hz, 2H), 6.92 (d,
J¼9.0 Hz, 2H), 5.06 (dd, J¼5.9 Hz, J¼6.2 Hz, 1H, H-2), 4.99 (dd,
J¼6.2 Hz, J¼7.2 Hz, 1H, H-20), 4.82 (d, J¼5.3 Hz, 1H, H-1), 4.66 (d,
J¼6.0 Hz, 1H, H-10), 4.29 (dd, J¼6.6 Hz, J¼11.9 Hz, 1H, H-5a), 4.03
(dd, J¼4.4 Hz, J¼11.9 Hz, 1H, H-5a0), 3.91 (dd, J¼3.4 Hz, J¼6.6 Hz, 1H,
H-4), 3.81 (s, 3H), 3.78 (t, J¼7.2 Hz,1H, H-30), 3.70 (m, 2H, H-3, H-40),
3.58 (dd, J¼3.2 Hz, J¼12.1 Hz, 1H, H-5b0), 3.28 (dd, J¼7.9 Hz,
J¼11.6 Hz, 1H, H-5b), 2.57 (d, J¼6.0 Hz, 1H, OH), 2.31 (s, 3H), 2.06 (s,
3H), 0.96 (t, J¼8.0 Hz, 9H), 0.88 (t, J¼8.0 Hz, 9H), 0.63 (q, J¼8.1 Hz,
in 5.0 mL CH2Cl2 was added DMAP (50 mg, 0.41 mmol),
EDCI(0.068 mg, 0.35 mmol) and cinnamic acid, after stirring for
24 h at room temperature, the solution was concentrated in vac-
uum, then the residue was purified by column chromatography on
silica gel (petroleum ether:ethyl acetate¼10:1) to afford compound
21 (0.16 g, 61%). 1H NMR (400 MHz, CDCl3):
d
7.67 (d, J¼16.0 Hz,1H),
7.51 (m, 2H), 7.39 (m, 3H), 7.24 (d, J¼8.6 Hz, 2H), 7.19 (d, J¼8.6 Hz,
2H), 6.85 (d, J¼8.6 Hz, 2H), 6.77 (d, J¼8.6 Hz, 2H), 6.35 (d, J¼16.0 Hz,
1H), 5.06 (t, J¼8.6 Hz, 1H, H-2), 4.73 (d, AB, J¼11.3 Hz, 1H), 4.65 (d,
AB, J¼12.0 Hz, 2H), 4.55 (d, AB, J¼11.3 Hz, 1H), 4.48 (d, J¼8.9 Hz, 1H,
6H), 0.55 (q, J¼8.1 Hz, 6H). 13C NMR (100 MHz, CDCl3):
d 169.5,
165.3, 163.5, 137.4, 132.0(2ꢂ Ar), 131.8(2ꢂ Ar), 131.4, 129.6(2ꢂ Ar),

4098
C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
122.0, 113.6(2ꢂ Ar), 101.5(C-10), 86.3(C-1), 75.0(C-4), 74.1(C-30),
73.6(C-20), 72.1(C-2), 70.8(C-40), 69.4(C-3), 65.0(C-50), 63.1(C-5),
55.4(eOCH3), 21.0(AreCH3), 20.9(CH3eCOe), 6.8(eCH2eCH3),
5.0(eCH2eCH3), 4.9(eCH2eCH3). C39H60O11SSi2, HRMS (ESI): calcd
for (MþNaþ): 815.3293 found: 815.3288.
55.3, 55.1, 21.0, 21.0. C44H48O12S, HRMS (ESI): calcd for (MþNaþ):
823.2764; found: 823.2757.
4.4.12. 3
tyldimethylsilyl)
thylsilyl)- -xylopyranosyl-(1/4)-2-O-acetyl-3-O-triethylsilyl-
arabinopyranoside] (38)/16 ,22(S),26-tetrahydroxycholest-5-ene-
3,26-di-(tert-butyldimethylsilyl) 16-O-[2-O-(4-methoxybenzoyl)-3,4-
di-(O-triethylsilyl)- -xylopyranosyl-(1/3)-2-O-acetyl-4-O-trie-
thylsilyl- -arabinop-yranoside] (39)/3 ,16 ,22(S),26-
tetrahydroxycholest-5-ene-3,26-di-(tert-butyldimethylsilyl) 16-O-{2-
O-[4-(E)-cinnamoyl]-3,4-di-(O-p-methoxybenzyl)- -xylopyr-
anosyl-(1/4)-2-O-acetyl-3-O-triethylsilyl- -arabinopyranoside}
(40)/3 ,16 ,22(S),26-tetrahydroxycholest-5-ene-3,26-di-(tert-bu-
tyldimethylsilyl) 16-O-{2-O-[4-(E)-cinnamoyl]-3,4-di-(O-p-methox-
ybenzyl)- -xylopyranosyl-(1/3)-2-O-acetyl-4-O-triethylsilyl-
arabinopyranoside} (41)/3 ,16 ,26-tetrahydroxycholest-5-ene-3,26-
di-(tert-butyldimethylsilyl) 16-O-{2-O-[4-(E)-cinnamoyl]-3,4-di-(O-
p-methoxybenzyl)- -xylopyranosyl-(1/4)-2-O-acetyl-3-O-trie-
thylsilyl- -arabinopyranoside} (42)/3
5-ene-3,26-di-(tert-butyldimethylsilyl)
moyl]-3,4-di-(O-p-methoxybenzyl)-
acetyl-4-O-triethylsilyl- -arabinopyranoside} (43). The mixture of
b
,16
b
,22(S),26-Tetrahydroxycholest-5-ene-3,26-di-(tert-bu-
16-O-[2-O-(4-methoxybenzoyl)-3,4-di-(O-trie-
(31) 1H NMR (300 MHz, CDCl3):
d
8.05 (d, J¼8.9 Hz, 2H), 7.11 (d,
b
-
D
a-L-
J¼7.9 Hz, 2H), 7.00 (d, J¼7.9 Hz, 2H), 6.89 (d, J¼8.9 Hz, 2H), 5.16 (dd,
J¼3.4 Hz, J¼4.3 Hz, 1H, H-2), 5.06 (t, J¼5.9 Hz, 1H, H-20), 4.93 (d,
J¼3.2 Hz, 1H, H-1), 4.84 (d, J¼5.6 Hz, 1H, H-10), 4.12 (dd, J¼7.8 Hz,
J¼10.7 Hz, 1H, H-5a), 4.05 (dd, J¼4.3 Hz, J¼12.6 Hz, 1H, H-5a0), 3.99
(dd, J¼3.8 Hz, J¼6.6 Hz,1H, H-3), 3.97 (m,1H, H-4), 3.85 (s, 3H), 3.83
(t, J¼7.1 Hz, 1H, H-30), 3.69 (m, 1H, H-40), 3.58 (dd, J¼4.1 Hz,
J¼11.4 Hz, 1H, H-5b), 3.35 (dd, J¼6.8 Hz, J¼11.8 Hz, 1H, H-5b0), 2.97
(d, J¼8.6 Hz, 1H, OH), 2.27 (s, 3H), 1.97 (s, 3H), 0.94 (t, J¼7.8 Hz, 9H),
0.90 (t, J¼7.8 Hz, 9H), 0.60 (q, J¼7.8 Hz, 6H), 0.60 (q, J¼7.8 Hz, 6H).
b
b-D
a
-
L
b
b
b-D
a-L
b
b
13C NMR (100 MHz, CDCl3):
d
169.4, 164.8, 163.3, 137.1, 132.0(2ꢂ Ar),
b
-
D
a-L-
131.8(2ꢂ Ar), 131.6, 129.4(2ꢂ Ar), 122.8, 113.4(2ꢂ Ar), 99.0(C-10),
85.7(C-1), 74.0(C-3), 73.1(C-30), 72.0(C-20), 70.4(C-40), 70.2(C-2),
64.6(C-4), 64.3(C-50), 62.2(C-5), 55.3(eOCH3), 21.0(AreCH3),
20.8(CH3eCOe), 6.7(eCH2eCH3), 4.9(eCH2eCH3), 4.8(eCH2eCH3).
b
b
b-D
a-L
b,16b
,26-tetrahydroxycholest-
16-O-{2-O-[4-(E)-cinna-
C
39H60O11SSi2, HRMS (ESI): calcd for (MþNaþ): 815.3293 found:
815.3299.
b-D-xylopyranosyl-(1/3)-2-O-
a-L
4.4.11. 4-Methylphenyl {2-O-[4-(E)-cinnamoyl]-3,4-di-(O-p-methox-
compound 34 (0.40 g, 0.42 mmol) and 8 (0.31 g, 0.46ꢀmmol) was
ꢀ
ybenzyl)-
binopyranoside}(32)/4-methylphenyl {2-O-[4-(E)-cinnamoyl]-3,4-di-
(O-p-methoxybenzyl)- -xylopyranosyl-(1/3)-2-O-acetyl-4-hy-
droxy-1- -arabinopyranoside} (33). The mixture of compound 23
b-
D
-xylopyranosyl-(1/4)-2-O-acetyl-3-hydroxy-1-
a-
L
-ara-
added 4 A MS (1.00 g) in dried CH2Cl2 under Ar at -60 C, then the
TMSOTf (6.60 mL) was slowly added, after stirring for 3 h, Et3N was
b
-
D
added, MS was filtered and filtrate was concentrated, then the
residue was purified by column chromatography on silica gel (pe-
troleum ether:ethyl acetate¼10:1) to afford compound 38 (0.40 g,
64%). Following the same procedure 39(58%), 40(51%), 41(47%),
42(55%), 43(43%) were achieved. (38) 1H NMR (400 MHz, CDCl3):
a-L
ꢀ
(8.40 g, 0.013 mol) and 29 (3.40 g, 0.011 mol) was added 4 A MS in
dried CH2Cl2 under Ar at ꢁ60 ꢀC, then the BF3$Et2O (0.077 mL,
0.061 mmol) was slowly added, after stirring for 3 h, Et3N was
added, MS was filtered and filtrate was concentrated, then the
residue was purified by column chromatography on silica gel (pe-
troleum ether:ethyl acetate¼8:1) to afford compound 32 (2.40 g,
d
8.03 (d, J¼8.7 Hz, 2H), 6.90 (d, J¼8.7 Hz, 2H), 5.30 (d, J¼4.9 Hz, 1H,
H-6), 4.96 (t, J¼6.6, 1H, H-20), 4.70 (dd, J¼7.0 Hz, J¼9.0 Hz,1H, H-200),
4.66 (d, J¼7.0 Hz, 1H, H-100), 4.33 (m, 1H), 4.09e3.95 (m, 3H), 3.86 (s,
3H), 3.72 (m, 4H), 3.46 (m, 4H), 3.33 (dd, J¼6.9 Hz, J¼9.0 Hz, 1H),
3.25 (dd, J¼8.6 Hz, J¼11.9 Hz, 1H), 2.28e2.20 (m, 2H), 1.97 (s, 3H),
1.81e1.70 (m, 2H), 0.64e0.44 (m, 18H), 0.06e0.02 (m, 12H). 13C
21%) and 33 (1.50 g, 13%). (32) 1H NMR (400 MHz, CDCl3):
d 7.65 (d,
J¼16.0 Hz,1H), 7.51 (m, 2H), 7.39 (m, 3H), 7.35 (d, J¼8.0 Hz, 2H), 7.25
(d, J¼8.4 Hz, 2H), 7.18 (d, J¼8.4 Hz, 2H), 7.07 (d, J¼8.0 Hz, 2H), 6.87
(d, J¼8.5 Hz, 2H), 6.78 (d, J¼8.5 Hz, 2H), 6.33 (d, J¼16.0 Hz, 1H), 5.06
(t, J¼6.4 Hz, 1H, H-2), 5.01 (t, J¼7.4 Hz, 1H, H-20), 4.78 (d, J¼5.9 Hz,
1H, H-1), 4.73 (d, J¼11.3 Hz, 1H), 4.64 (dd, J¼7.0 Hz, J¼11.1 Hz, 2H),
4.61 (d, J¼6.6 Hz, 1H, H-10), 4.54 (d, J¼11.3 Hz, 1H), 4.25 (dd,
J¼5.8 Hz, J¼12.1 Hz, 1H, H-5a), 3.97 (dd, J¼3.5 Hz, J¼12.6 Hz, 1H, H-
5a0), 3.93 (m, 1H, H-4), 3.80 (s, 3H), 3.74 (m, 1H, H-3), 3.67 (s, 3H),
3.64 (m, 2H, H-30, H-40), 3.56 (dd, J¼2.5 Hz, J¼11.9 Hz, 1H, H-5b),
3.29 (dd, J¼8.2 Hz, J¼11.4 Hz, 1H, H-5b0), 2.70 (m, 1H, OH), 2.23 (s,
NMR (100 MHz, CDCl3):
d
168.8, 163.7, 163.2, 141.4, 131.7(2ꢂ
AreCH), 122.6, 120.9, 113.3(2ꢂ AreCH), 101.7, 101.0, 81.6, 77.1, 74.8,
74.1, 73.4, 73.1, 72.6, 71.7, 71.1, 68.4, 65.5, 65.0, 57.1, 55.3, 54.8, 51.1,
42.7, 42.0, 39.6, 37.2, 36.4, 36.2, 35.8, 34.9, 32.0, 31.8, 31.4, 30.5, 29.6,
25.9(3ꢂ Me3CSi), 25.9(3ꢂ Me3CSi), 22.6, 20.9, 19.3, 18.3, 18.2, 16.8,
12.7, 11.2, 6.8(6ꢂ CH2CH3), 6.7(3ꢂ CH2CH3), 4.9(3ꢂ CH2CH3), 4.9(3ꢂ
CH2CH3), 4.5(3ꢂ CH2CH3), ꢁ4.6(2ꢂ Me2Si), ꢁ5.4(2ꢂ Me2Si).
C77H140O15Si5, HRMS (ESI): calcd for (MþNaþ): 1467.8936; found:
1467.8945.
3H), 2.05 (s, 3H). 13C NMR (100 MHz, CDCl3):
d 169.8, 165.8, 159.4,
159.2, 145.9, 137.6, 134.2, 132.3, 130.9, 130.8, 130.5, 130.2, 130.0,
129.7(2ꢂ Ar), 129.6(2ꢂ Ar), 129.5(2ꢂ Ar), 128.8(2ꢂ Ar), 128.3(2ꢂ
Ar), 117.2, 113.9(2ꢂ Ar), 113.7(2ꢂ Ar), 101.7(C-10), 86.3(C-1), 79.6(C-
40), 76.8(C-30), 75.3(C-4), 74.0, 72.8, 72.6(C-20), 72.0(C-2), 70.1(C-3),
64.2(C-5), 63.5(C-50), 55.3, 55.1, 21.1, 20.9. C44H48O12S, HRMS (ESI):
calcd for (MþNaþ): 823.2764; found: 823.2773.
(39) 1H NMR (400 MHz, CDCl3):
d
8.02 (d, J¼8.8 Hz, 2H), 6.91 (d,
J¼8.8 Hz, 2H), 5.32 (d, J¼5.1 Hz, 1H, H-6), 4.96 (m, 1H), 4.90 (m, 1H),
4.81 (m, 1H), 4.32 (m, 1H), 4.21 (m, 1H), 4.03 (m, 2H), 3.91 (m, 1H),
3.87 (s, 3H), 3.78 (m, 2H), 3.60 (m, 1H), 3.45 (m, 3H), 3.27 (m, 3H),
1.92 (s, 3H), 0.64e0.55 (m, 18H), 0.07e0.04 (m, 12H). 13C NMR
(150 MHz, CDCl3):
d
168.6, 164.5, 163.2, 141.6, 141.5, 131.8(2ꢂ
(33) 1H NMR (400 MHz, CDCl3):
d
7.65 (d, J¼16.0 Hz,1H), 7.46 (m,
AreCH),122.7,121.0,121.0,113.3(2ꢂ AreCH),101.4, 100.8, 81.8, 78.5,
75.6, 74.5, 72.6, 72.6, 71.9, 70.2, 68.3, 68.0, 65.5, 63.0, 57.0, 55.3,
54.8, 50.1, 42.8, 42.1, 40.0, 39.4, 37.3, 36.5, 36.2, 35.9, 35.2, 32.0, 31.8,
31.5, 30.3, 25.9(3ꢂ Me3CSi), 25.9(3ꢂ Me3CSi), 20.8, 20.7, 19.2, 18.3,
18.2, 16.9, 13.0, 11.7, 6.9(3ꢂ CH2CH3), 6.8(3ꢂ CH2CH3), 6.8(3ꢂ
CH2CH3), 4.9(6ꢂ CH2CH3), 4.8(3ꢂ CH2CH3), ꢁ4.6(2ꢂ Me2Si), ꢁ5.3,
ꢁ5.4. C77H140O15Si5, HRMS (ESI): calcd for (MþNaþ): 1467.8936;
found: 1467.8945.
2H), 7.37 (m, 3H), 7.21 (m, 4H), 6.92 (d, J¼8.0 Hz, 2H), 6.86 (d,
J¼8.4 Hz, 2H), 6.77 (d, J¼8.4 Hz, 2H), 6.39 (d, J¼16.0 Hz, 1H), 5.17 (t,
J¼3.5 Hz, 1H, H-2), 5.11 (t, J¼7.1 Hz, 1H, H-20), 4.93 (d, J¼3.5 Hz, 1H,
H-1), 4.73 (d, J¼12.0 Hz, 1H), 4.65 (d, J¼10.9 Hz, 2H), 4.65 (d,
J¼7.1 Hz, 1H, H-10), 4.54 (d, J¼12.0 Hz, 1H), 4.09 (m, 1H, H-5a0), 3.98
(dd, J¼3.8 Hz, J¼11.2 Hz, 1H, H-5a), 3.95 (m, 2H, H-3, H-4), 3.79 (s,
3H), 3.67 (s, 3H), 3.64 (m, 2H, H-30, H-40), 3.55 (dd, J¼3.8 Hz,
J¼11.2 Hz, 1H, H-5b), 3.33 (m, 1H, H-5b0), 2.23 (s, 3H), 2.03 (s, 3H).
(40) 1H NMR (400 MHz, CDCl3):
d
7.64 (d, J¼16.0 Hz,1H), 7.51 (m,
13C NMR (100 MHz, CDCl3):
d
169.5, 165.5, 159.5, 159.2, 145.2, 137.2,
2H), 7.38 (m, 3H), 7.23 (d, J¼8.4 Hz, 2H), 7.23 (d, J¼8.4 Hz, 2H), 6.85
(d, J¼8.4 Hz, 2H), 6.74 (d, J¼8.4 Hz, 2H), 6.36 (d, J¼16.0 Hz, 1H), 5.29
(d, J¼4.2 Hz, 1H, H-6), 5.01 (t, J¼8.0 Hz, 1H), 4.72e4.51 (m, 6H), 4.36
(m, 1H), 4.30 (t, J¼6.6 Hz, 1H), 4.09e3.94 (m, 3H), 3.79 (s, 3H),
3.86e3,75 (m, 2H), 3.64 (s, 3H), 3.59 (m, 2H), 3.45 (m, 2H), 3.26 (m,
134.4, 131.7(2ꢂ Ar), 131.7, 130.3, 130.1, 129.9, 129.9 (2ꢂ Ar),
129.5 (2ꢂ Ar), 128.8 (2ꢂ Ar), 128.2 (2ꢂ Ar), 118.1, 113.9 (2ꢂ Ar),
113.7 (2ꢂ Ar), 100.4(C-10), 85.7(C-1), 79.3(C-30), 76.6(C-40), 75.9(C-
3), 74.0, 72.9, 71.3(C-20), 70.8(C-2), 64.8(C-4), 63.5(C-50), 62.5(C-5),

C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
4099
2H),1.99 (s, 3H), 0.52 (q, J¼7.9 Hz, 6H), 0.06e0.01 (m,12H). 13C NMR
3.86 (s, 3H), 3.70 (m, 4H), 3.45 (m, 4H), 3.24 (dd, J¼8.0 Hz, J¼11.7 Hz,
1H), 2.87 (m, 1H), 2.61 (m, 1H),1.95 (s, 3H), 0.54 (m,18H), 0.09e0.01
(75 MHz, CDCl3):
d 168.8, 165.2, 159.3, 159.0, 145.0, 141.4, 134.4,
130.2, 130.2, 129.9, 129.8(2ꢂ AreCH), 129.4(2ꢂ AreCH), 128.8(2ꢂ
AreCH), 128.0(2ꢂ AreCH), 120.9, 117.8, 113.8(2ꢂ AreCH), 113.6(2ꢂ
AreCH),102.0,100.8, 80.1, 77.2, 75.9, 75.6, 74.9, 73.9, 73.9, 73.2, 72.7,
72.6, 72.2, 69.1, 68.4, 63.3, 57.1, 55.2, 54.9, 54.8, 50.1, 42.7, 42.0, 39.6,
37.2, 36.5, 36.2, 35.7, 34.8, 32.0, 31.8, 31.4, 30.3, 29.6, 25.9(6ꢂ
Me3CSi), 22.6, 20.9, 19.3, 18.3, 18.2, 16.8, 12.8, 11.1, 6.8(3ꢂ CH2CH3),
4.6(3ꢂ CH2CH3), ꢁ4.6(2ꢂ Me2Si), ꢁ5.4(2ꢂ Me2Si). C82H128O16Si3,
HRMS (ESI): calcd for (MþNaþ): 1475.8408; found: 1475.8423.
(m, 12H). 13C NMR (75 MHz, CDCl3):
d 214.3, 168.8, 164.6, 163.2,
141.4, 131.7, 122.8, 121.0, 113.4, 101.7, 100.1, 82.6, 80.5, 74.6, 73.5,
73.1, 72.6, 72.3, 71.1, 68.1, 65.0, 61.7, 56.4, 55.3, 54.1, 53.1, 50.0, 43.8,
42.8, 41.4, 39.6, 38.8, 37.3, 36.5, 35.4, 35.3, 31.9, 31.7, 31.3, 29.3,
25.9(6ꢂ Me3CSi), 22.7, 22.6, 20.8, 19.4, 18.3, 18.2, 16.6, 13.3, 6.8(9ꢂ
CH2CH3), 5.0(3ꢂ CH2CH3), 4.9(3ꢂ CH2CH3), 4.6(3ꢂ CH2CH3),
ꢁ4.6(2ꢂ Me2Si), ꢁ5.4(2ꢂ Me2Si). C77H138O15Si5, HRMS (ESI): calcd
for (MþNaþ): 1465.8780; found: 1465.8790.
(41) 1H NMR (400 MHz, CDCl3):
d
7.65 (d, J¼16.0 Hz,1H), 7.50 (m,
(45) 1H NMR (400 MHz, CDCl3):
d
8.04 (d, J¼8.3 Hz, 2H), 6.90 (d,
2H), 7.37 (m, 3H), 7.23 (d, J¼8.5 Hz, 2H), 7.23 (d, J¼8.5 Hz, 2H), 6.81
(d, J¼8.5 Hz, 2H), 6.75 (d, J¼8.5 Hz, 2H), 6.37 (d, J¼16.0 Hz, 1H), 5.29
(d, J¼4.2 Hz, 1H, H-6), 5.00 (m, 1H), 4.71e4.54 (m, 6H), 4.28 (m, 1H),
4.10 (m, 1H), 3.97 (m, 3H), 3.75 (s, 3H), 3.70 (m, 1H), 3.68 (s, 3H),
3.59 (m, 2H), 3.45 (m, 3H), 3.33 (m, 2H), 2.02 (s, 3H), 0.60 (q,
J¼8.0 Hz, 6H), 0.06e0.01 (m, 12H). C82H128O16Si3, HRMS (ESI): calcd
for (MþNaþ): 1475.8408; found: 1475.8415.
J¼8.3 Hz, 2H), 5.30 (d, J¼4.4 Hz, 1H, H-6), 4.91 (m, 1H), 4.80 (m, 2H),
4.2 (m, 1H), 4.07 (m, 1H), 3.93 (m, 2H), 3.86 (s, 3H), 3.85 (m, 1H),
3.76 (m, 2H), 3.60 (m, 1H), 3.46 (m, 1H), 3.37 (m, 1H), 3.25 (m, 3H),
2.84 (m, 1H), 2.42 (m, 1H), 1.93 (s, 3H), 0.56 (m, 18H), 0.05e0.00 (m,
12H). 13C NMR (100 MHz, CDCl3):
d 214.1, 168.9, 164.5, 163.5, 141.4,
131.9(2ꢂ AreCH), 122.4, 113.5(2ꢂ AreCH), 99.6, 98.6, 80.7, 74.2,
72.6, 71.9, 71.4, 69.6, 69.2, 68.2, 64.7, 63.1, 61.1, 56.1, 55.3, 54.1, 50.0,
43.7, 42.8, 41.4, 39.6, 38.0, 37.2, 36.5, 35.3, 35.1, 32.0, 31.7, 31.3, 29.6,
26.9, 25.9(6ꢂ Me3CSi), 20.7, 20.7, 19.3, 18.3, 18.2, 16.3, 16.2, 13.3,
6.8(6ꢂ CH2CH3), 6.7(3ꢂ CH2CH3), 4.8(6ꢂ CH2CH3), 4.7(3ꢂ CH2CH3),
ꢁ4.6(2ꢂ Me2Si), ꢁ5.4(2ꢂ Me2Si). C77H138O15Si5, HRMS (ESI): calcd
for (MþNaþ): 1465.8780; found: 1465.8785.
(42) 1H NMR (400 MHz, CDCl3):
d
7.65 (d, J¼16.0 Hz,1H), 7.50 (m,
2H), 7.37 (m, 3H), 7.23 (d, J¼8.2 Hz, 2H), 7.18 (d, J¼8.2 Hz, 2H), 6.85
(d, J¼8.2 Hz, 2H), 6.75 (d, J¼8.2 Hz, 2H), 6.38 (d, J¼16.0 Hz, 1H), 5.30
(m, 1H, H-6), 5.01 (t, J¼6.8 Hz, 1H), 4.78e4.51 (m, 6H), 4.30 (m, 1H),
4.02e3.93 (m, 2H), 3.85e3.71 (m, 3H), 3.79 (s, 3H), 3.65 (s, 3H), 3.62
(m, 2H), 3.52e3.39 (m, 3H), 3.29 (m, 2H), 1.98 (s, 3H), 0.53 (q,
J¼7.92 Hz, 6H), 0.05 (s, 6H), 0.01 (s, 6H). C82H128O15Si3, HRMS (ESI):
calcd for (MþNaþ): 1459.8459; found: 1459.8464.
(46) 1H NMR (400 MHz, CDCl3):
d
7.63 (d, J¼16.0 Hz,1H), 7.50 (m,
2H), 7.39 (m, 3H), 7.23 (d, J¼8.0 Hz, 2H), 7.16 (d, J¼8.0 Hz, 2H), 6.85
(d, J¼7.8 Hz, 2H), 6.74 (d, J¼7.8 Hz, 2H), 6.34 (d, J¼16.0 Hz, 1H), 5.31
(d, J¼3.4 Hz, 1H, H-6), 4.98 (t, J¼7.64 Hz, 1H), 4.72e4.51 (m, 6H),
4.09 (m, 1H), 3.95 (m, 3H), 3.80 (m, 1H), 3.79 (s, 3H), 3.70 (m, 2H),
3.64 (s, 3H), 3.56 (m, 2H), 3.47 (m, 2H), 3.32 (m, 1H), 3.25 (m, 1H),
1.98 (s, 3H), 0.54 (q, J¼8.0 Hz, 6H), 0.06e0.01 (m, 12H). 13C NMR
(43) 1H NMR (400 MHz, CDCl3):
d
7.64 (d, J¼16.0 Hz,1H), 7.51 (m,
2H), 7.37 (m, 3H), 7.23 (d, J¼8.5 Hz, 2H), 7.20 (d, J¼8.5 Hz, 2H), 6.83
(d, J¼8.5 Hz, 2H), 6.75 (d, J¼8.5 Hz, 2H), 6.35 (d, J¼16.0 Hz, 1H), 5.27
(d, J¼3.5 Hz, 1H, H-6), 5.06 (t, J¼6.8 Hz, 1H), 4.99 (t, J¼6.2 Hz, 1H),
4.69e4.51 (m, 6H), 4.30 (m, 1H), 4.20 (m, 1H), 3.96 (m, 1H), 3.85 (m,
2H), 3.77 (s, 3H), 3.67 (s, 3H), 3.60 (m, 2H), 3.45e3.29 (m, 5H), 1.98
(s, 3H), 0.58 (m, 6H), 0.04 (s, 6H), 0.01 (s, 6H). 13C NMR (100 MHz,
(100 MHz, CDCl3):
d 214.2, 168.8, 165.2, 159.3, 159.1, 145.0, 141.3,
134.3, 130.2, 130.1, 130.0, 129.8(2ꢂ AreCH), 129.5(2ꢂ AreCH),
128.8(2ꢂ AreCH), 128.0(2ꢂ AreCH), 120.9, 117.8, 113.8(2ꢂ AreCH),
113.6(2ꢂ AreCH), 102.1, 100.0, 80.1, 77.1, 75.7, 74.9, 73.9, 73.0, 72.7,
72.6, 72.2, 68.1, 66.7, 63.3, 56.4, 55.2, 55.0, 54.1, 50.0, 43.7, 42.8, 42.7,
41.3, 39.6, 38.8, 37.2, 36.5, 35.3, 32.0, 31.3, 30.5, 27.2, 25.9(3ꢂ
Me3CSi), 25.9(3ꢂ Me3CSi), 22.6, 20.9, 19.3, 18.2, 18.2, 16.5, 16.3, 13.3,
6.8(3ꢂ CH2CH3), 4.6(3ꢂ CH2CH3), ꢁ4.6(2ꢂ Me2Si), ꢁ5.4(2ꢂ Me2Si).
CDCl3):
d 168.7, 165.5, 159.5, 159.2, 145.1, 141.6, 134.7, 130.6, 130.4,
130.3, 129.6(2ꢂ AreCH), 129.6(2ꢂ AreCH), 129.0(2ꢂ AreCH),
128.3(2ꢂ AreCH), 121.3, 118.1, 114.0(2ꢂ AreCH), 113.7(2ꢂ AreCH),
102.9, 100.6, 81.9, 77.1, 76.2, 75.0, 73.5, 72.8, 72.7, 72.7, 68.7, 68.7,
65.7, 60.8, 60.6, 55.4, 55.4, 55.3, 55.2, 50.5, 43.0, 42.3, 39.8, 37.5,
36.7, 36.5, 36.3, 36.1, 34.4, 32.3, 31.9, 31.5, 30.8, 29.9, 26.2(3ꢂ
Me3CSi), 26.1(3ꢂ Me3CSi), 21.3, 21.0, 19.6, 18.6, 18.5, 17.0, 14.4, 13.0,
7.0(3ꢂ CH2CH3), 5.0(3ꢂ CH2CH3), ꢁ4.3(2ꢂ Me2Si), ꢁ5.0, ꢁ5.1.
C
82H126O16Si3, HRMS (ESI): calcd for (MþNaþ): 1473.8251; found:
1473.8256.
(47) 1H NMR (400 MHz, CDCl3):
d
7.67 (d, J¼16.0 Hz,1H), 7.50 (m,
C
82H128O15Si3, HRMS (ESI): calcd for (MþNaþ): 1459.8459; found:
2H), 7.38 (m, 3H), 7.22 (d, J¼8.0 Hz, 2H), 7.20 (d, J¼8.0 Hz, 2H), 6.81
(d, J¼8.6 Hz, 2H), 6.74 (d, J¼8.6 Hz, 2H), 6.42 (d, J¼16.0 Hz, 1H), 5.28
(d, J¼3.4 Hz, 1H, H-6), 5.04 (t, J¼5.5 Hz, 1H), 4.70e4.52 (m, 6H), 4.06
(m, 1H), 3.89 (m, 3H), 3.78 (m, 1H), 3.75 (s, 3H), 3.70 (m, 1H), 3.67 (s,
3H), 3.58 (m, 2H), 3.45 (m, 2H), 3.35 (m, 2H), 3.29 (m, 1H), 2.88 (m,
1H), 2.63 (m, 1H), 2.03 (s, 3H), 0.60 (m, 6H), 0.07e0.01 (m, 12H).
1459.8457.
4.4.13. 3
tyldimethylsilyl)
thylsilyl)- -xylopyranosyl-(1/4)-2-O-acetyl-3-O-triethylsilyl-
arabinopyranoside] (44)/16 ,26-trihydroxycholest-5-en-22-one-
3,26-di-(tert-butyldimethylsilyl) 16-O-[2-O-(4-methoxybenzoyl)-3,4-
di-(O-triethylsilyl)- -xylopyranosyl-(1/3)-2-O-acetyl-4-O-trie-
thylsilyl- -arabinopyranoside] (45)/16
22-one-3,26-di-(tert-butyldimethylsilyl)
moyl]-3,4-di-(O-p-methoxybenzyl)-
acetyl-3-O-triethylsilyl- -arabinopyranoside} 46)/16
trihydroxycholest-5-en-22-one-3,26-di-(tert-butyldimethylsilyl) 16-
b
,16
b
,26-Trihydroxycholest-5-en-22-one-3,26-di-(tert-bu-
16-O-[2-O-(4-methoxybenzoyl)-3,4-di-(O-trie-
b
-
D
a-L-
b
C
82H126O16Si3, HRMS (ESI): calcd for (MþNaþ): 1473.8251; found:
1473.8259.
b-D
a-
L
b
,26-trihydroxycholest-5-en-
16-O-{2-O-[4-(E)-cinna-
4.4.14. 3
[O-2-O-(4-methoxybenzoyl)-
-arabinopyranoside] (48a)/3
5-en-22-hydroxy 16-O-[O-2-O-(4-methoxybenzoyl)-
anosyl-(1/4)-2-O-acetyl- -arabinopyranoside] (48b)/3
trihydroxycholest-5-en-22-one 16-O-[O-2-O-(4-methoxybenzoyl)-
-xylopyranosyl-(1/3)-2-O-acetyl- -arabinopyranoside] (49a)/
,16 ,26-trihydroxycholest-5-en-22-one 16-O-[O-2-O-(4-
methoxybenzoyl)- -xylopyranosyl-(1/4)-2-O-acetyl- -arabino-
b
,16
b
,22(S),26-Tetrahydroxycholest-5-en-22-hydroxy 16-O-
-xylopyranosyl-(1/3)-2-O-acetyl-
,16 ,22(S),26-tetrahydroxycholest-
-xylopyr-
,16 ,26-
b-D
b
-
D
-xylopyranosyl-(1/4)-2-O-
,26-
a-L
b
b
a
-
L
b
b-D
a
-
L
b
b
O-{2-O-[4-(E)-cinnamoyl]-3,4-di-(O-p-methoxybenzyl)-
a n o s yl - ( 1 /3 ) - 2 - O - a c e t yl - 4 - O - t r i e t hyl s i l yl -
b
-
a
D
-xylopyr-
- a ra -
b-
-
L
D
a-L
binopyranoside}(47). A solution of 38 (0.087 g, 0.060 mmol) in
CH2Cl2 (3.00 mL), was added PDC (0.069 g, 0.18 mmol), after stirring
for 6 h at room temperature, the concentrate was purified by col-
umn chromatography on silica gel (petroleum ether:ethyl
acetate¼10:1) to afford compound 44 (0.084 g, 96%). Following the
same procedure 45 (67%), 46 (80%), 47 (74%) were achieved. (44) 1H
3
b
b
b
-D
a-L
pyranoside] (49b). To a solution of 38 (0.021 g, 0.014 mmol) in
acetone/H2O (1 mL, 10: 1) was added Pd(CH3CN)2Cl2 (0.0030 g).
After the reaction was stirred at room temperature for 3 h, the
solvent was removed. The residue was isolated by silica gel column
chromatography (petroleum ether:ethyl acetate¼12:1), then puri-
fied by Sephadex LH-20 column chromatography (chloroform:
MeOH¼1:1) to afford 48b (0.0087 g, 67%). Following the same
NMR (400 MHz, CDCl3):
2H), 5.32 (d, J¼4.1 Hz, 1H, H-6), 4.94 (t, J¼6.7 Hz, 1H), 4.66 (d,
J¼6.1 Hz, 1H), 4.58 (m, 1H), 4.07 (m, 1H), 3.99 (m, 2H), 3.88 (m, 1H),
d
8.00 (d, J¼8.8 Hz, 2H), 6.89 (d, J¼8.8 Hz,

4100
C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
procedure 48a (yield: 59%), 49a (yield: 62%), 49b (yield: 57%) were
achieved. (48a) 1H NMR (600 MHz, CD3OD):
8.04 (d, J¼8.8 Hz, 2H),
was removed. The residue was isolated by silica gel column chro-
matography (petroleum ether:ethyl acetate¼12:1), then purified by
Sephadex LH-20 column chromatography (chloroform:MeOH¼1:1)
to afford 50b (6.70 mg, 54%). Following the same procedure 50a
(yield: 59%), 51a (yield: 52%), 51b (yield: 53%), 52a (yield: 48%), 52b
(yield: 61%) were achieved. (50a) 1H NMR (600 MHz, CD3OD):
d
7.03 (d, J¼8.8 Hz, 2H), 5.34 (d, J¼4.9 Hz, 1H, H-6), 5.03 (dd, J¼7.2 Hz,
J¼9.4 Hz, 1H, H-20), 4.94 (dd, J¼7.3 Hz, J¼9.0 Hz, 1H, H-200), 4.72 (d,
J¼7.3 Hz, 1H, H-100), 4.29 (d, J¼7.2 Hz, 1H, H-10), 4.14 (m, 1H, H-16),
4.05 (m, 1H, H-40), 3.98 (m, 1H, H-500), 3.89 (s, 3H), 3.89 (dd,
J¼3.0 Hz, J¼12.5 Hz, 1H, H-50), 3.80 (dd, J¼2.1 Hz, J¼9.4 Hz, 1H, H-
30), 3.64 (m, 2H, H-300, H-400), 3.55 (dd, J¼1.3 Hz, J¼12.5 Hz,1H, H-50),
3.44 (m, 1H, H-26), 3.43 (m, 1H, H-22), 3.37 (m, 1H, H-3), 3.35 (m,
1H, H-500), 3.32 (m, 1H, H-26), 2.25 (m, 2H, H-4), 2.17 (m, 2H, H-15),
1.97 (m, 2H, H-11), 1.87 (m, 1H, H-1), 1.8 (m, 2H, H-2), 1.59 (s, 3H),
1.52 (m, 3H, H-7, H-25), 1.48 (m, 2H, H-12), 1.47 (m, 1H, H-17), 1.40
(m, 2H, H-23), 1.31 (m, 1H, H-8), 1.07 (m, 1H, H-1), 1.02 (s, 3H, H-19),
0.91 (m, 1H, H-9), 0.89 (d, J¼6.6 Hz, 3H, H-27), 0.87 (m, 1H, H-14),
0.86 (d, J¼7.0 Hz, 3H, H-21), 0.81 (s, 3H, H-18). 13C NMR (150 MHz,
d
7.75 (d, J¼16.0 Hz, 1H), 7.65 (m, 2H), 7.44 (m, 3H), 6.57 (d,
J¼16.0 Hz, 1H), 5.34 (d, J¼5.3 Hz, 1H, H-6), 5.07 (dd, J¼7.1 Hz,
J¼9.3 Hz, 1H, H-20), 4.84 (m, 1H, H-200), 4.64 (d, J¼7.4 Hz, 1H, H-100),
4.27 (d, J¼7.1 Hz,1H, H-10), 4.15 (m, 1H, H-16), 4.0 (m,1H, H-40), 3.96
(dd, J¼5.2 Hz, J¼11.7 Hz,1H, H-500), 3.87 (dd, J¼3.0 Hz, J¼12.6 Hz,1H,
H-50), 3.79 (dd, J¼3.3 Hz, J¼9.3 Hz, 1H, H-30), 3.62 (m, 1H, H-400),
3.54 (dd, J¼1.3 Hz, J¼12.6 Hz, 1H, H-50), 3.53 (t, J¼8.7 Hz, 1H, H-300),
3.48 (m, 1H, H-22), 3.41 (m, 1H, H-26), 3.38 (m, 1H, H-3), 3.30 (m,
2H, H-500, H-26), 2.23 (m, 2H, H-4), 2.19 (m, 1H, H-15), 2.00 (s, 3H),
1.98 (m, 1H, H-11), 1.84 (m, 1H, H-15), 1.82 (m, 1H, H-20), 1.79 (m,
2H, H-2), 1.62 (m, 2H, H-24), 1.53 (m, 1H, H-25), 1.52 (m, 1H, H-23),
1.50 (m, 2H, H-7, H-17), 1.44 (m, 3H, H-12, H-25), 1.32 (m, 1H, H-8),
1.31 (m, 1H, H-23), 1.05 (m, 2H, H-1), 1.02 (s, 3H, H-19), 0.92 (m, 1H,
H-9), 0.91 (d, J¼6.7 Hz, 3H, H-21), 0.89 (d, J¼6.6 Hz, 3H, H-27), 0.87
(m, 1H, H-14), 0.84 (s, 3H, H-18). 13C NMR (150 MHz, CD3OD):
CD3OD):
d
170.9, 167.1, 165.4, 142.2(C-5), 133.2(2ꢂ Ar), 123.4,
122.3(C-6), 114.7(2ꢂ Ar), 103.6(C-100), 103.1(C-10), 81.4(C-16),
81.0(C-30), 75.6(C-300), 75.1(C-200), 73.9(C-22), 72.4(C-20), 72.2(C-3),
70.9(C-400), 69.6(C-40), 68.3(C-26), 66.5(C-500), 66.3(C-50), 58.7(C-
17), 56.3(C-14), 56.0(eOCH3), 51.7(C-9), 43.1(C-13), 42.9(C-4),
41.0(C-11), 38.4(C-1), 37.6(C-10), 37.2(C-25), 37.2(C-15), 36.5(C-20),
33.9(C-23), 32.8(C-7), 32.2(C-2), 31.6(C-24), 30.6(C-8), 21.8(C-12),
20.9, 19.8(C-19), 17.5(C-27), 13.1(C-18), 11.6(C-21). C47H70O15, HRMS
(ESI): calcd for (MþNaþ): 897.4612; found: 897.4610.
d
169.4, 166.3, 145.3, 140.7(C-5), 134.3, 130.2, 129.4, 128.7, 127.9,
120.9(C-6), 117.5, 102.2(C-100), 101.8(C-10), 80.1(C-16), 79.7(C-30),
74.3(C-300), 73.5(C-200), 72.5(C-22), 71.0(C-3), 71.0(C-20), 69.3(C-400),
68.3(C-40), 66.8(C-26), 65.2(C-500), 65.1(C-50), 57.4(C-17), 55.1(C-14),
50.2(C-9), 41.7(C-13), 41.5(C-4), 39.6(C-11), 37.0(C-1), 36.4(C-10),
35.8(C-25), 35.2(C-15), 35.1(C-20), 31.6(C-23), 31.6(C-7), 30.8(C-2),
30.2(C-24), 29.4(C-8), 22.3(C-12), 20.1, 18.4(C-19), 16.1(C-27),
11.7(C-18), 10.2(C-21). C48H70O14, HRMS (ESI): calcd for (MþNaþ):
893.4663; found: 893.4660.
(48b) 1H NMR (600 MHz, CD3OD):
d
8.05 (d, J¼8.9 Hz, 2H), 6.98
(d, J¼8.9 Hz, 2H), 5.36 (d, J¼4.7 Hz, 1H, H-6), 4.96 (dd, J¼7.4 Hz,
J¼9.1 Hz, 1H, H-200), 4.85 (d, J¼7.4 Hz, 1H, H-100), 4.78 (dd, J¼5.5 Hz,
J¼7.9 Hz, 1H, H-20),4.31 (d, J¼5.5 Hz, 1H, H-10), 4.11 (m, 1H, H-16),
4.00 (dd, J¼5.0 Hz, J¼12.0 Hz, 1H, H-50), 3.93 (m, 1H, H-400), 3.87 (s,
3H), 3.69 (dd, J¼3.3 Hz, J¼7.9 Hz, 1H, H-30), 3.65 (m, 1H, H-40), 3.64
(m, 1H, H-300), 3.53 (dd, J¼2.4 Hz, J¼12.0 Hz, 1H, H-50), 3.50 (m, 1H,
H-22), 3.46 (m, 1H, H-26), 3.40 (m, 1H, H-3), 3.33 (m, 2H, H-26, H-
500), 2.25 (m, 2H, H-4), 2.15 (m, 1H, H-15), 2.00 (m, 2H, H-11), 2.00 (s,
3H), 1.87 (m, 2H, H-1, H-20), 1.80 (m, 2H, H-2), 1.56 (m, 2H, H-24, H-
25), 1.52 (m, 4H, H-7, H-12), 1.50 (m, 1H, H-17), 1.42 (m, 2H, H-23),
1.31 (m, 3H, H-8, H-24), 1.07 (m, 1H, H-1), 1.05 (s, 3H, H-19), 0.93 (d,
J¼6.6 Hz, 3H, H-27), 0.92 (m, 1H, H-9), 0.90 (d, J¼7.0 Hz, 3H, H-21),
0.86 (m, 1H, H-14), 0.82 (s, 3H, H-18). 13C NMR (150 MHz, CD3OD):
(50b) 1H NMR (600 MHz, CD3OD):
d
7.76 (d, J¼16.0 Hz, 1H), 7.60
(m, 2H), 7.41 (m, 3H), 6.62 (d, J¼16.0 Hz, 1H), 5.34 (d, J¼4.8 Hz, 1H,
H-6), 4.87 (m, 2H, H-20, H-200), 4.81 (d, J¼7.6 Hz, 1H, H-100), 4.32 (d,
J¼6.0 Hz, 1H, H-10), 4.14 (m, 1H, H-16), 3.99 (dd, J¼4.5 Hz, J¼12.1 Hz,
1H, H-50), 3.97 (m, 1H, H-500), 3.95 (m, 1H, H-400), 3.74 (dd, J¼3.3 Hz,
J¼8.2 Hz, 1H, H-30), 3.63 (m, 1H, H-40), 3.58 (m, 1H, H-300), 3.54 (dd,
J¼2.2 Hz, J¼12.1 Hz, 1H, H-50), 3.52 (m, 1H, H-22), 3.47 (m, 1H, H-
26), 3.41 (m, 1H, H-3), 3.35 (m, 1H, H-26), 3.30 (dd, J¼1.5 Hz,
J¼11.7 Hz, 1H, H-500), 2.24 (m, 2H, H-4), 2.19 (m, 1H, H-15), 1.98 (m,
1H, H-11), 1.97 (s, 3H), 1.88 (m, 1H, H-20), 1.86 (m, 1H, H-1), 1.81 (m,
1H, H-2), 1.63 (m, 1H, H-8), 1.61 (m, 1H, H-24), 1.56 (m, 1H, H-25),
1.53 (m, 2H, H-12), 1.51 (m, 1H, H-17), 1.50 (m, 2H, H-7), 1.48 (m, 2H,
H-23), 1.14 (m, 1H, H-24), 1.04 (s, 3H, H-19), 1.01 (m, 1H, H-1), 0.93
(d, J¼6.7 Hz, 3H, H-27), 0.92 (m, 1H, H-9), 0.87 (m, 1H, H-14), 0.86
(d, J¼7.0 Hz, 3H, H-21), 0.79 (s, 3H, H-18). 13C NMR (150 MHz,
d
171.3, 167.7, 165.1, 142.2(C-5), 133.1(2ꢂ Ar), 123.5, 122.3(C-6),
114.6(2ꢂ Ar), 103.5(C-100), 102.6(C-10), 82.0(C-16), 77.1(C-400),
75.8(C-300), 75.7(C-200), 74.1(C-22), 73.9(C-20), 72.4(C-3), 71.1(C-30),
71.0(C-40), 68.4(C-26), 66.7(C-500), 64.3(C-50), 58.6(C-17), 56.2(C-
14), 55.9(eOCH3), 51.7(C-9), 43.1(C-4), 43.0(C-13), 41.0(C-11),
38.4(C-1), 37.6(C-10), 37.3(C-25), 37.0(C-15), 36.5(C-20), 33.9(C-23),
32.8(C-7), 32.2(C-2), 31.6(C-24), 30.6(C-8), 21.9(C-12), 20.9, 19.8(C-
19), 17.4(C-27), 13.3(C-18), 11.6(C-21). C47H70O15, HRMS (ESI): calcd
for (MþNaþ): 897.4612; found: 897.4615.
CD3OD):
d 171.3, 168.2, 146.7, 142.2(C-5), 136.0, 131.3, 129.9, 129.4,
122.3(C-6), 119.1, 103.5(C-100), 102.8(C-10), 81.7(C-16), 77.1(C-400),
75.8(C-300), 75.5(C-20), 74.1(C-200), 73.9(C-22), 72.4(C-3), 71.4(C-30),
71.1(C-40), 68.4(C-26), 66.8(C-500), 64.9(C-50), 58.7(C-17), 56.3(C-14),
51.7(C-9), 43.1(C-4), 43.0(C-13), 41.0(C-11), 38.4(C-1), 37.6(C-10),
37.3(C-25), 37.1(C-15), 36.5(C-20), 32.9(C-23), 32.8(C-7), 32.2(C-2),
31.6(C-24), 30.6(C-8), 21.9(C-12), 21.1, 19.9(C-19), 17.4(C-27), 13.2(C-
4.4.15. 3
{O-2-O-[4-(E)-cinnamoyl]-
arabinopyranoside} (50a)/3
22-hydroxy 16-O-{O-2-O-[4-(E)-cinnamoyl]-
(1/4)-2-O-acetyl- -arabinopyranoside} (50b)/3
trihydroxycholest-5-en-22-one 16-O-{O-2-O-[4-(E)-cinnamoyl]-
xylopyranosyl (1/3)-2-O-acetyl- -arabinopyranoside} (51a)/
,16 ,26-trihydroxycholest-5-en-22-one 16-O-{O-2-O-[4-(E)-cinna-
moyl]-
(51b)/3
moyl]-
(52a)/3
moyl]-
b
,16
b
,22(S),26-Tetrahydroxycholest-5-en-22-hydroxy 16-O-
-xylopyranosyl (1/3)-2-O-acetyl-
,16 ,22(S),26-tetrahydroxycholest-5-en-
-xylopyranosyl
,16 ,26-
b
-
D
a-L-
b
b
b-D
a
-
L
b
b
18), 11.6(C-21).
893.4663; found: 893.4665.
(51a) 1H NMR (600 MHz, CD3OD):
C
48H70O14
,
HRMS (ESI): calcd for (MþNaþ):
b-D-
a-L
d
7.77 (d, J¼16.0 Hz, 1H), 7.66
3b
b
(m, 2H), 7.45 (m, 3H), 6.60 (d, J¼16.0 Hz, 1H), 5.34 (d, J¼4.9 Hz, 1H,
H-6), 4.98 (dd, J¼6.9 Hz, J¼9.1 Hz, 1H, H-20), 4.84 (m, 1H, H-200), 4.62
(d, J¼7.6 Hz, 1H, H-100), 4.06 (d, J¼6.9 Hz, 1H, H-10), 4.02 (m, 1H, H-
16), 3.97 (m, 1H, H-500), 3.96 (m, 1H, H-40), 3.85 (dd, J¼3.1 Hz,
J¼12.5 Hz, 1H, H-50), 3.76 (dd, J¼3.2 Hz, J¼9.1 Hz, 1H, H-30), 3.62 (m,
1H, H-400), 3.56 (t, J¼8.6 Hz, 1H, H-300), 3.49 (dd, J¼1.6 Hz, J¼12.5 Hz,
1H, H-50), 3.40 (m, 1H, H-3), 3.35 (m, 2H, H-26), 3.31 (m, 1H, H-500),
2.85 (m, 1H, H-20), 2.65 (m, 1H, H-23), 2.45 (m, 1H, H-23), 2.24 (m,
2H, H-4), 2.14 (m, 1H, H-15), 2.07 (s, 3H), 1.93 (m, 2H, H-11), 1.85 (m,
2H, H-1), 1.8 (m, 2H, H-2), 1.69 (m,1H, H-17),1.59 (m,1H, H-24),1.57
(m, 1H, H-12), 1.52 (m, 2H, H-7), 1.50 (m, 1H, H-12), 1.48 (m, 1H, H-
b
-
D
-xylopyranosyl (1/4)-2-O-acetyl-
a
-
L
L
-arabinopyranoside}
-arabinopyranoside}
b
,16
b
,26-trihydroxycholest-5-en-16-O-{O-2-O-[4-(E)-cinna-
b-
D-xylopyranosyl (1/3)-2-O-acetyl-
a-
b
,16
b
,26-trihydroxycholest-5-en-16-O-{O-2-O-[4-(E)-cinna-
-arabinopyranoside}
b-
D
-xylopyranosyl (1/4)-2-O-acetyl-a-L
(52b). To a solution 40 (0.021 g, 0.014 mmol) in CH2Cl2/H2O (1 mL,
10:1) was added DDQ (9.5 mg, 0.043 mmol). The reaction was
stirred at ambient temperature for 5 h, then CH2Cl2 was removed,
and acetone (1 mL) and Pd(CH3CN)2Cl2 (0.0030 g) was added. After
the reaction was stirred at room temperature for 3 h, the solvent

C. Liu et al. / Tetrahedron 72 (2016) 4091e4102
4101
25), 1.32 (m, 1H, H-8), 1.27 (m, 1H, H-24), 1.09 (d, J¼7.2 Hz, 3H, H-
21), 1.03 (s, 3H, H-19), 0.95 (m, 1H, H-9), 0.92 (m, 1H, H-14), 0.85 (d,
J¼6.7 Hz, 3H, H-27), 0.83 (s, 3H, H-18). 13C NMR (150 MHz, CD3OD):
3.20 (m, 1H, H-500), 2.19 (m, 2H, H-4), 2.05 (s, 3H), 1.97 (m, 1H, H-11),
1.80 (m, 1H, H-1), 1.76 (m, 2H, H-2), 1.54 (m, 1H, H-25), 1.47 (m, 2H,
H-7), 1.46 (m, 3H, H-12, H-20), 1.40 (m, 2H, H-22), 1.31 (m, 4H, H-15,
H-23), 1.28 (m, 1H, H-8), 1.11 (m, 2H, H-11, H-17), 1.02 (m, 3H, H-1,
H-24),1.00 (s, 3H, H-19), 0.93 (d, J¼7.5 Hz, 3H, H-27), 0.88 (m,1H, H-
9), 0.81 (m, 1H, H-14), 0.81 (d, J¼5.4 Hz, 3H, H-21). 13C NMR
d
216.4(C-22), 170.9, 167.8, 146.8, 142.2(C-5), 135.7, 131.6, 130.8,
130.1, 1209.3, 122.2(C-6), 118.9, 103.7(C-100), 102.1(C-10), 80.9(C-16),
80.7(C-3 ), 75.8(C-300), 75.0(C-200), 72.5(C-20), 72.3(C-3), 70.9(C-400),
69.4(C-500), 68.1(C-26), 66.7(C-40), 66.1(C-50), 57.6(C-17), 55.5(C-14),
51.6(C-9), 44.9(C-20), 42.9(C-4), 42.8(C-13), 41.0(C-11), 39.2(C-23),
38.4(C-1), 37.6(C-10), 36.6(C-15), 36.3(C-25), 32.7(C-7), 32.2(C-2),
30.4(C-8), 28.0(C-24), 21.8(C-12), 21.5, 19.8(C-19), 17.0(C-27),
16.9(C-21), 13.5(C-18). C48H68O14, HRMS (ESI): calcd for (MþNaþ):
891.4507; found: 891.4510.
(150 MHz, CD3OD):
d 170.1, 167.6, 144.8, 140.7(C-5), 130.0, 128.5,
127.9, 127.7, 120.9(C-6), 117.1, 105.3(C-100), 102.6(C-10), 81.4(C-16),
78.4(C-400), 76.3(C-300), 73.6(C-200), 73.5(C-3), 73.0(C-20), 70.9(C-30),
70.7(C-40), 69.5(C-26), 65.2(C-500), 64.4(C-50), 60.6(C-17), 54.8(C-
14), 50.2(C-9), 41.8(C-13), 41.5(C-4), 39.5(C-11), 37.0(C-1), 36.2(C-
10), 36.0(C-15), 35.9(C-22), 35.4(C-25), 33.8(C-24), 31.4(C-20),
31.3(C-7), 30.8(C-2), 29.2(C-8), 22.2(C-23), 20.4(C-12), 19.9, 18.3(C-
19), 17.2(C-27), 15.7(C-18), 11.7(C-21). C48H70O13, HRMS (ESI): calcd
for (MþNaþ): 877.4714; found: 877.4716.
(51b) 1H NMR (600 MHz, CD3OD):
d
7.77 (d, J¼16.0 Hz, 1H), 7.64
(m, 2H), 7.42 (m, 3H), 6.62 (d, J¼16.0 Hz, 1H), 5.34 (d, J¼4.9 Hz, 1H,
H-6), 4.86 (m, 1H, H-200), 4.80 (d, J¼7.6 Hz, 1H, H-100), 4.77 (dd,
J¼5.7 Hz, J¼8.0 Hz, 1H, H-20), 4.09 (d, J¼5.7 Hz, 1H, H-10), 4.01 (m,
1H, H-16), 3.95 (dd, J¼4.9 Hz, J¼11.5 Hz, 1H, H-500), 3.94 (dd,
J¼4.6 Hz, J¼11.9 Hz, 1H, H-50), 3.91 (m, 1H, H-400), 3.72 (dd, J¼3.1 Hz,
J¼8.0 Hz, 1H, H-30), 3.61 (m, 1H, H-40), 3.58 (t, J¼8.9 Hz, 1H, H-300),
3.49 (dd, J¼1.9 Hz, J¼11.9 Hz, 1H, H-50), 3.4 (m, 1H, H-3), 3.34 (m,
2H, H-26), 3.30 (m, 1H, H-500), 2.88 (m, 1H, H-20), 2.70 (m, 1H, H-
23), 2.55 (m,1H, H-23), 2.25 (m, 2H, H-4), 2.12 (m,1H, H-15), 2.02 (s,
3H), 1.94 (m, 1H, H-11), 1.86 (m, 1H, H-1), 1.81 (m, 2H, H-2), 1.72 (m,
1H, H-17), 1.62 (m, 1H, H-24), 1.53 (m, 1H, H-25), 1.52 (m, 2H, H-12),
1.52 (m, 2H, H-7), 1.32 (m, 1H, H-8), 1.30 (m, 1H, H-24), 1.23 (m, 1H,
H-11), 1.10 (d, J¼7.3 Hz, 3H, H-21), 1.04 (s, 3H, H-19), 0.95 (m, 1H, H-
9), 0.92 (m, 1H, H-14), 0.90 (d, J¼6.7 Hz, 3H, H-27), 0.80 (s, 3H, H-
Acknowledgements
This work was supported by the National Program of New Drug
Innovation and Production (2009ZX09301-003-9-1) and Beijing
Key Laboratory of Active Substances Discovery and Drug Ability
Evaluation. And we are grateful to Dr. Wanqi Zhou for the antitu-
mor activity test and thanks Dr. Yanshen Guo provide molecule
simulation test for this work.
Supplementary data
18). 13C NMR (150 MHz, CD3OD):
d 216.6(C-22), 171.4, 168.3, 146.7,
Supplementary data (1H NMR, 13C NMR, COSY, HSQC, HMBC,
NOESY, HPLC spectra of target compounds.) associated with this
article can be found in the online version, at http://dx.doi.org/
10.1016/j.tet.2016.05.049.
142.2(C-5), 136.0, 131.4, 130.8, 129.9, 129.4, 122.2(C-6), 119.0,
103.5(C-100), 101.8(C-10), 81.3(C-16), 77.0(C-400), 75.8(C-300), 75.5(C-
200), 74.0(C-20), 72.3(C-3), 71.1(C-30), 71.1(C-40), 68.2(C-26), 66.8(C-
500), 64.4(C-50), 57.7(C-17), 55.5(C-14), 51.5(C-9), 44.8(C-20), 42.9(C-
4), 42.7(C-13), 41.0(C-11), 39.3(C-23), 38.4(C-1), 37.6(C-10), 36.4(C-
15), 36.4(C-25), 32.8(C-7), 32.2(C-2), 30.6(C-8), 28.0(C-24), 21.9(C-
References and notes
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d
7.74 (d, J¼16.0 Hz, 1H), 7.65
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d 169.5, 166.3, 145.3, 140.8(C-5), 134.3, 130.2,
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33.9(C-24), 31.5(C-20), 31.4(C-7), 30.8(C-2), 29.2(C-8), 22.1(C-23),
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877.4708.
(52b) 1H NMR (600 MHz, CD3OD):
d
7.72 (d, J¼16.0 Hz, 1H), 7.59
(m, 2H), 7.38 (m, 3H), 6.50 (d, J¼16.0 Hz, 1H), 5.30 (d, J¼4.8 Hz, 1H,
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3.88 (m, 1H, H-400), 3.70 (m, 1H, H-30), 3.54 (m, 1H, H-300), 3.50 (m,
1H, H-50), 3.46 (m, 2H, H-26), 3.34 (m, 1H, H-40), 3.30 (m, 1H, H-3),
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