Synthesis and biological evaluation of novel Jiyuan Oridonin A-1,2,3-triazole-azole derivatives as antiproliferative agents

Article abstract of DOI:10.1016/j.ejmech.2018.08.056

As a continuation of our research on developing potent and potentially safe anti-proliferative agents, two series of novel Jiyuan Oridonin A-1,2,3-triazole-azole hybrids were designed, synthesized and evaluated for their anti-proliferative activity against four selected cancer cell lines (MGC-803, MCF-7, PC-3, Eca-109). Some compounds with better growth inhibitory effects were chosen to carry out further studies in A549 and SMMC-7721. Most of the synthesized compounds exhibited moderate to good activity against all the cancer cell lines selected. Particularly, the most active agent 8b showed high potency against human cancer cells with IC₅₀ ranging from 0.2 ± 0.0 to 5.0 ± 0.9 μM. Cellular mechanism studies elucidated compound 8b arrests cell cycle at G1 phase and induce a strong apoptotic response in SMMC-7721 cells. Furthermore, 8b could inhibit the colony formation and migration via Wnt signaling pathway in SMMC-7721 cells. For all these reasons, compound 8b holds promising potential as anti-proliferative agent.

Full text of DOI:10.1016/j.ejmech.2018.08.056

Accepted Manuscript
Synthesis and biological evaluation of novel Jiyuan Oridonin A-1,2,3-triazole-azole
derivatives as antiproliferative agents
Yu Ke, Jian-Jia Liang, Rui-Juan Hou, Ming-Ming Li, Long-Fei Zhao, Wang Wang,
Ying Liu, Hang Xie, Rui-Hua Yang, Tian-Xing Hu, Jin-Yi Wang, Hong-Min Liu
PII:
S0223-5234(18)30726-8
10.1016/j.ejmech.2018.08.056
EJMECH 10667
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 17 June 2018
Revised Date: 20 August 2018
Accepted Date: 20 August 2018
Please cite this article as: Y. Ke, J.-J. Liang, R.-J. Hou, M.-M. Li, L.-F. Zhao, W. Wang, Y. Liu, H. Xie, R.-
H. Yang, T.-X. Hu, J.-Y. Wang, H.-M. Liu, Synthesis and biological evaluation of novel Jiyuan Oridonin
A-1,2,3-triazole-azole derivatives as antiproliferative agents, European Journal of Medicinal Chemistry
(2018), doi: 10.1016/j.ejmech.2018.08.056.
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ACCEPTED MANUSCRIPT
A focused library of Jiyuan Oridonin A-1,2,3-triazole-azole derivatives were designed
and synthesis. Compound 8b displayed the most potent inhibition against
SMMC-7721 cells, inhibited colony formation and migration of SMMC-7721, as well
as induced apoptosis.

ACCEPTED MANUSCRIPT
Synthesis and biological evaluation of novel Jiyuan Oridonin
A-1,2,3-triazole-azole derivatives as antiproliferative agents
1
1
Yu Ke , Jian-Jia Liang , Rui-Juan Hou, Ming-Ming Li, Long-Fei Zhao, Wang Wang,
Ying Liu, Hang Xie, Rui-Hua Yang, Tian-Xing Hu, Jin-Yi Wang, Hong-Min Liu*
School of Pharmaceutical Sciences; Institute of Drug Discovery and Development;
Key Laboratory of Advanced Pharmaceutical Technology, Ministry of Education of
China; Zhengzhou University, Zhengzhou 450001, PR China.
Collaborative Innovation Center of New Drug Research and Safety Evaluation,
Henan Province, Zhengzhou 450001, PR China.
1
These authors contributed equally to this work.
Corresponding Authors:
Prof. Hong-Min Liu
Zhengzhou University, Zhengzhou, 450001, China
E-mail: liuhm@zzu.edu.cn (Hong-Min Liu)
Abstract
As a continuation of our research on developing potent and potentially safe
anti-proliferative agents, two series of novel Jiyuan Oridonin A-1,2,3-triazole-azole
hybrids were designed, synthesized and evaluated for their anti-proliferative activity
against four selected cancer cell lines (MGC-803, MCF-7, PC-3, Eca-109). Some
compounds with better growth inhibitory effects were chosen to carry out further
studies in A549 and SMMC-7721. Most of the synthesized compounds exhibited
moderate to good activity against all the cancer cell lines selected. Particularly, the
most active agent 8b showed high potency against human cancer cells with IC50
ranging from 0.2 ± 0.0 to 5.0 ± 0.9 µM. Cellular mechanism studies elucidated
compound 8b arrests cell cycle at G1 phase and induce a strong apoptotic response in
SMMC-7721 cells. Furthermore, 8b could inhibit the colony formation and migration
via Wnt signaling pathway in SMMC-7721 cells. For all these reasons, compound 8b
holds promising potential as anti-proliferative agent.
1

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Keywords
:
ent-kaurene diterpenoid; 1,2,3-triazole-azole; Apoptosis; Wnt signaling
pathway
1
. Introduction
Cancer, being one of the leading causes of death globally, causes almost 8
million people death each year and poses a major socioeconomic hazard to humanity
at large [1]. Although great advances have been made in the field of cancer prevention
and treatment over the past few decades, the success to treat cancer remains a
challenge [2]. Therefore, development of novel anticancer agents with broader
spectrum of cytotoxicity to fight cancer are urgently needed.
Natural products with unique chemical skeletons and low toxicity are an
important source of new drugs, especially in the discovery of anticancer compounds.
Over 60% anticancer drugs have their origin from natural sources [3]. Isodon species
are widely distributed plants, and more than 600 diterpenoids have been isolated from
this species over the past 40 years [4]. Some of them showed distinct pharmacological
activities and low toxicity, especially ent-kaurane diterpenoids with exo-methylene
cyclopentanone in the D-ring such as Oridonin, Ponicidin and Phyllostachysin F, and
so on [5, 6] (Fig. 1). Isodon rubescens (“Donglingcao” in Chinese), a widely
distributed ent-kaurene in the Isodon plants, has attracted growing attention in recent
years due to its extensive and unique pharmacological effects as antibacterial [7],
anti-tumor and anti-inflammatory agents [8, 9]. Increasing studies have also illustrated
that Oridonin exerts broad anti-tumor activities through regulating the cell cycle [10,
11], apoptosis [12], and autophagy [13], as well as the inhibition of migration and
invasion [14]. The unique skeleton and potent antitumor activity makes them
promising candidates as antitumor agents [15]. However, the development of Isodon
diterpenoids for cancer therapy was impeded largely by their relatively moderate
bioavailability [16]. Therefore, it is particularly urgent to develop novel derivatives by
rational modification to enhance its potency and antitumor activity profile [17].
Previous studies have revealed that the modified of heterocycle-fused ring A and
1
4-hydroxy group could markedly increase the anti-proliferative efficacy of Oridonin
2

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and its analogues [18]. For instance, C. Ding reported that it could significantly
enhance the potency and aqueous solubility toward cancer cells to fuse the thiazole
with Oridonin A-ring system [19] (Fig. 1). However, there are few reports toward
modifications of the ent-kaurane diterpenoid’s 20- positions to generate diversity.
Fig. 1. Structures of ent-kaurene diterpenoids from Isodon and some representative Oridonin
incorporated with heterocycle derivatives have been reported.
1,2,3-triazole is one of the crucial structural units found in a large number of
bioactive molecules as anti-fungal [20], anti-bacterial [21-23], anti-allergic [24],
anti-HIV [25], anti-tubercular [26], and anti-inflammatory agents [27]. Moreover, this
heterocycle has a high dipole moment and is capable of hydrogen bonding, which
could be favorable in the binding of bio-molecular targets [28]. In recent years, people
are increasingly focused on their anticancer activity [29-32]. On the other hand, the
heterocyclic azoles, including 1,3,4-thiadiazole-2-thiol, 1-methyl-1H-tetrazole-5-thiol,
4
,5-dihydrothiazole-2-thiol, benzo[d]thiazole-2-thiol and 1,3-thiazolidine-2,4-dione,
represent one of the most effective cores of anticancer agents, with a wide range of
activities against various cancer cell lines [2, 33, 34].
3

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Our previous studies focused on another ent-kaurene diterpenoid Jiyuan
Oridonin A (The following is abbreviated JOA, Fig. 1), which was isolated from
Isodon rubescens by our group. It showed more potent in inhibiting proliferation and
pro-apoptotic in various human cancer cell lines than Oridonin, such as colon cancer,
gastric cancer and esophageal cancer cell lines [35, 36]. Furthermore, the JOA's
2
0-acetal derivatives containing 1,2,3-trizole group exhibited better anticancer activity
than JOA itself in vitro and in vivo [37, 38]. These above encouraging findings and
our continuous quest to identify more potent analogues, led to the molecular
hybridization of JOA, 1,2,3-trizole, and different azoles to generate a new architecture
with the aim of exploring the impact of such modification on the anticancer agents
and making better use of the special scaffold of JOA. To our delight, most of the
synthesized compounds exhibited robust activity against all the cancer cell lines
selected. Moreover, the anticancer mechanisms of our compounds involved Wnt
signaling pathway. These results provide new insights to the potential use of Isodon
diterpenoids as novel and efficient anticancer agents. The detailed illustration of target
derivatives was shown in Fig 2.
Fig. 2. Illustration of the design strategy for target compounds.
2
. Results and discussion
2
.1. Chemistry
The general route for the synthesis of the target Jiyuan Oridonin A
-1,2,3-triazole-azole analogues 5a-5f was depicted in Scheme 1. Compound 1 mixed
with 3-bromoethanol catalyzed by p-TsOH in THF at room temperature to provide
4

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compound 2, and then reacted with NaN in the system of ethyl acetoacetate and
3
water (5:1) to afford compound 3. Commercially available azoles were treated with
propargyl bromide to provide 4a-4f, which were further reacted with 3 by click
reaction to afford compounds 5a-5f with good yields.
Compound 5e showed better anti-proliferative activities than other compounds as
the anti-proliferative activities of series 1 were tested. It was supposed that thiazole
groups may improve the anticancer activities of compounds. Besides, modifications of
the 1,3-thiazolidine-2,4-dione have proven highly effective with regards to improved
potency and less toxicity [17, 39]. On this basis, another series of compounds bearing
a thiazolidinedione(TZD) ring were designed and synthesized according to Scheme 2.
Similar to the first route, compound 3 could be obtained directly following the first
procedures. Commercially available 2,4-thiazolidinedione was treated with propargyl
bromide to provide compound 6 which then undergoes Knoevenagal condensation
with different aldehydes to yield 7a-7k, which were subjected to click reaction with
compound 3 to yield compounds 8a-8k. Additionally, we also synthesized compound
1
6
a for comparison. All the synthesized compounds were characterized by H NMR,
1
3
C NMR, HRMS and IR.
Scheme 1. Synthetic route of series 1. Reagents and conditions: i) 3-Bromo-1-propanol, THF,
p-TsOH, reflux, 3h; ii) EtOAC/H O, NaN , reflux, 7h; iii) alkyne intermediates, CuSO /Cu,
2
3
4
t-BuOH/H O, reflux, 8-12h.
2
5

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Scheme 2. Synthetic route of series 2. Reagents and conditions: i) 3-Bromopropyne, K CO ,
2
3
acetone, reflux, 2h; ii) appropriate aldehyde, piperidine, 4-6h; iii) alkyne intermediates, CuSO /Cu,
4
t-BuOH/H O, reflux, 8-12h.
2
2
.2. Biological evaluation
2
.2.1. In Vitro antiproliferative activity and SAR study
The anti-proliferative activities against four human cancer cell lines, including
Eca-109 (human esophageal cancer cell line), MGC-803 (human stomach cancer line),
MCF-7 (human breast cancer cell line), PC-3 (human prostate cancer cell line), were
determined for all the prepared compounds, and the cytotoxic activity of target
compounds were detected by MTT assay. The IC values (concentration required to
5
0
inhibit tumor cell proliferation by 50%) were listed in Table 1 and Table 2, with
Oridonin and JOA used as positive control.
From the screening results in Table 1, it was observed that all the compounds
exhibited stronger anti-proliferative activities against all four selected cancer lines
than Oridonin. During the SAR (structure activity relationship) studies, we found that
5
e and 5f had a remarkable effect on their anti-proliferative activities by bearing
thiazole or benzothiazole ring with JOA (compound 5a-5f). And compound 5e
showed the most active inhibitory effect against the four-tested human cancer cell
6

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lines ranging from 0.8 to 2.6 µM, respectively. These results suggested that the
thiazole groups would be helpful to improve the anti-proliferative activities of the
ent-kaurene diterpenoids
a
Table 1. Inhibitory activities of compounds 5a-5f against the tested cancer cell lines.
IC (µM)
5
0
Compd
R1
-
Eca-109
33.6±1.2
MGC-803
34.3±1.7
PC-3
MCF-7
Oridonin
43.9±2.2
64.3±1.1
1
(Jiyuan
-
-
25.2±1.1
10.1±0.6
27.1±0.5
9.7±0.8
24.3±0.7
10.0±0.9
21.1±2.2
7.6±0.1
Oridonin A)
3
5
5
a
8.6±0.7
5.6±0.5
5.0±0.4
6.2±0.6
0.8±0.0
1.4±0.2
8.9±0.3
5.2±0.5
5.2±0.5
4.5±0.4
1.7±0.2
2.1±0.3
11.6 ±0.8
9.6 ±0.8
5.9±0.6
7.7±0.7
1.4±0.1
1.6±0.7
13.6±0.1
15.2±0.9
14.9±0.9
14.6±0.8
2.6±0.4
b
5
c
5
d
5
e
5
f
4.2±0.8
a
MTT methods, cell lines were treated with target compounds for 72h.The average
value of the three independent experiments was expressed by mean ± SDs.
In order to complete the SAR study, compound
with
1
,2,3-triazole-thiazolidinedione ring of JOA (compound 6a) was synthesized (Scheme
7

ACCEPTED MANUSCRIPT
2
) and its anti-proliferative activity results was shown in Table 2. Unfortunately, when
conjugated with thiazolidinedione, the anti-proliferative activity of compound 6a
decreased (6a VS. 5e or 5f), especially on the Eca-109 and MGC-803 cells, which
showed weak or no cytotoxicity. And it is well-known that the major structural
determinant for anticancer activity of Oridonin is the presence of the α, β-unsaturated
ketone (enone) system in the D-ring, destruction of this enone system could
counteract its anticancer activity [8, 17, 40]. To further determine the affection of the
additional enone motif in the TZD ring, compounds 8a-8k were synthesized and their
inhibitory activities were shown in Table 2. Delightedly, compounds 8a-8d led to a
dramatic increase in activity toward Eca-109 and MGC-803 cell lines. Meanwhile, the
length and type of carbon chain (compound 8a-8d) had little effect on anticancer
activity of the substituent of TZD ring. However, changing the aliphatic substituent
(compound 8a-8d) to aromatic unit (compound 8e-8k) led to a decrease of the activity,
particularly on the MGC-803 and PC-3 cells. All these results indicated that the
substituent of TZD ring may play an important role in determining activity.
Table 2. Inhibitory activities of compounds 6a and 8a-8k against the tested cancer
a
cell lines.
IC50 (µM)
Compd
Oridonin
R2
Eca-109
33.6±1.2
MGC-803
PC-3
MCF-7
-
-
34.3±1.7
43.8±2.2
64.3±1.0
1
(Jiyuan
25.2±1.1
27.1±0.5
24.3±0.7
21.1±2.2
Oridonin A)
3
-
-
10.1±0.6
35.0±1.3
2.5±0.2
2.7±0.2
9.7±0.8
>50
10.0±0.9
5.8±0.5
8.9±0.9
5.0±0.8
7.6±0.1
9.5±1.0
4.1±0.6
1.5±0.2
6a
8a
8b
2.1±0.1
0.6±0.0
8
c
4.9±0.6
2.5±0.2
6.2±0.8
4.3±0.6
8

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8
d
4.2±0.5
4.9±0.5
1.2±0.0
5.3±0.7
1.9±0.3
6.6±0.5
8
e
20.0±1.1
13.2±1.1
8
f
3.3±0.4
2.6±0.3
35.0±1.4
25.9±1.3
16.1±1.2
18.4±1.3
4.8±0.3
7.9±0.6
8
8
g
h
5.4±0.6
9.3±0.8
4.7±0.5
27.8±1.3
17.3±1.1
29.7±0.7
13.2±1.1
11.5±1.1
13.4±1.1
4.9±0.5
8.9±0.9
15.7±0.7
8
i
8
j
8
k
11.8±0.9
10.1±0.9
13.9±0.3
4.1±0.6
a
MTT methods, cell lines were treated with target compounds for 72h.The average
value of the three independent experiments was expressed by mean ± SDs.
As can be seen in Table 3, some compounds with better growth inhibitory effects
were chosen to carry out further studies in A549 (human lung cancer cell line) and
SMMC-7721(hepatoma carcinoma cell line). Among them, compound 8b was
generally observed for stronger anticancer activities in all tested cell lines compared
with Oridonin. Therefore, compound 8b was further examined for possible
cytotoxicity against HL-7702 (normal liver cell line). As showed in Fig. 3A, the
inhibitory rate of compound 8b on HL-7702 cells (IC50 = 18.1±1.2 µM) was
significantly less than that of SMMC-7721 (IC50 = 0.2±0.0 µM) at the same
concentration. The results indicated that compound 8b had good selectivity between
cancer and normal cells. Due to a remarkable broad-spectrum activity against all
tested human cancer cell lines and the best activity against SMMC-7721 cell line,
compound 8b was chosen to be further investigated regarding its mechanism of
action.
a
Table 3. IC Values of part of compounds for A549 and SMMC-7721 cell line.
5
0
Compd
R
IC (µM)
50
9

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A549
SMMC-7721
30.1±2.2
Oridonin
-
-
-
25.4±2.5
16.4±1.3
13.6±1.8
1
3
14.6±1.5
12.0±2.6
5
e
4.0±0.6
5.8±0.8
3.5±0.5
1.8±0.0
5
f
8
8
a
7.8±0.9
3.5±0.7
3.4±0.5
0.2±0.0
b
8
c
9.2±0.9
4.8±0.7
5.7±0.8
5.3±0.7
8d
a
MTT methods, cell lines were treated with target compounds for 72h.The average
value of the three independent experiments was expressed by mean ± SDs.
2
.2.2. Effect of compound 8b on the proliferation of SMMC-7721 cells
Further evaluation on dose-dependent and time-dependent inhibitory effects of
active compound 8b was performed. As shown in Fig. 3B, with the increase of drug
concentration range from 0 to 128 µmol/L and the extension of time, the inhibitory
effect also enhanced. The results suggested that compound 8b could inhibit cell
proliferation in a time-dependent and dose-dependent manner.
Fig. 3. (A) Effects of compound 8b on cancer cell line and normal cell line. (B) Inhibitory
activity of compound 8b on SMMC-7721 cells at different time.
1
0

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2
.2.3. Clone assays
Based on the most excellent activity against SMMC-7721 cells, cell proliferation
inhibiting ability of compound 8b was measured using colony formation assay (Fig.
A and B). This assay tested each cell in the population to undergo extensive divisions
4
and also monitored the cells that had retained the capacity for producing colonies after
the treatment with cell death causing agents (radiations or chemotherapeutic agents)
[
41]. As shown in Fig. 4A and B, SMMC-7721 cells treated with 8b exhibited fewer
and smaller colonies compared to the control with a colony formation efficiency of
.7% at 2 µM, which indicated that compound 8b could significantly inhibit the
5
proliferation of SMMC-7721 cells in a concentration-dependent manner.
Fig. 4. (A) Representative images of SMMC-7721 cells colonies after treatment with various
concentrations for two weeks; (B) Quantitative analysis of the colony formation efficiency. The
results shown were representative of three independent experiments. **: p < 0.01 verse control.
2
.2.4. Cell cycle arrest effects of 8b on SMMC-7721 cells
Most of the anticancer agents inhibited cell proliferation through induction of
cell growth arrest. To identify whether the high anticancer effects of the hybrids were
caused by cell cycle accumulated at a certain phase, the effects of different
concentrations of compound 8b on cell cycle progression were examined with
SMMC-7721 cell line. Exposure of SMMC-7721 cells to compound 8b for 12 h
11

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resulted in an interference with cell cycle distribution. As shown in Fig. 5A and B, the
percentage of cells in the G1 phase after the treatment with compound 8b at various
concentrations (1, 2 and 4 µmol/L) were 45.8%, 54.9% and 75.7% when compared
with the control (36.5%), whereas the proportion of cells at S and G2/M phase
decreased significantly upon the application of 8b (p < 0.05 or p < 0.01). The results
suggested that 8b caused a clear G1 arrest pattern in a concentration and
time-dependent manner, with a concomitant decrease of cells in other phases of the
cell cycle.
2
.2.5. Apoptosis effects of 8b on SMMC-7721 cells
To get further insight into the mode of cell death induced by compound 8b,
Annexin V/PI staining was used to quantitatively measure the effect of 8b on
SMMC-7721 cells apoptosis. After treatment with compound 8b for 12h at indicated
concentrations (0, 0.5, 1, 2 µmol/L), SMMC-7721 cells were labeled with the two
dyes, and the resulting red (PI) and green (FITC) fluorescence was monitored by flow
cytometry. As shown in cytometry apoptosis diagram (Fig. 6A and B), the percentage
of apoptotic cells were significantly increased from 2.8% (DMSO control) to 29.2%
after treatment with enhancing concentrations of 8b.
Fig. 5. Effect of compound 8b on the cell cycle distribution of SMMC-7721 cells. Cells were
treated with different concentrations (0, 1, 2, 4 µmol/L) for 12 h. Then the cells were fixed and
stained with PI to analyze DNA content by flow cytometry. (A) Diagram of the cell cycle for each
phase based on flow cytometry. (B) The proportions of cells in G1, S, and G2/M phases were
1
2

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calculated. The data were analyzed by Student's t-test.
Fig. 6. Compound 8b induced apoptosis of SMMC-7721 cells. (A) SMMC-7721 cells apoptosis
was detected by Annexin V/PI on flow cytometry. Cells treated with 0, 0.5, 1 and 2 µM 8b for 24
h were collected and processed for analysis. (B) The proportion of apoptotic (Annexin V positive)
cells increased dependently with 8b treatment concentration.
2
.2.6. The effects on Wnt signaling pathway of 8b-treated SMMC-7721 cells
From above studies, we can conclude that compound 8b inhibited growth of
SMMC-7721 cells. We next investigated whether compound 8b could inhibit
migration of cancer cells. Herein, the monolayer of SMMC-7721 cells was scratched
and treated with 8b at various concentrations (0, 0.2 and 0.5 µM) for 24 h. The area of
the wound was measured at two different time points 0 and 24 h and the percentage
reduction in cell migration was assessed by recovered area of the scratch as compared
with 0 h post scratch. As shown in Fig. 7A and B, compound 8b inhibited migration
of SMMC-7721 cells even at low concentration (0.2 µM). It was observed that cell
migration was inhibited in a dose dependent manner (Fig. 7B).
The Wnt signaling pathway has been demonstrated to play a critical role in
numerous cellular processes including tissue homeostasis, cell proliferation, and cell
migration [42, 43]. Aberrant Wnt signaling has been linked to the formation and
metastasis of tumors [44]. Specifically, once activated, β-catenin disassembles from
complex comprised of APC and GSK3β and translocates into nuclear [45]. To
elucidate the potential mechanisms contributed to antimigration activity by the
compound 8b, several proteins (APC, p-GSK3β, β-catenin, Dvl2) related to Wnt
1
3

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signaling pathway were determined by Western blotting to confirm this property. As
shown in Fig. 7C and D, it is clearly to find the treatment of SMMC-7721 cells with
8
b resulted in the increase expression of APC and p-GSK3β, and the decrease
expression of β-catenin and Dvl2. These results indicated that compound 8b might
inhibit migration of SMMC-7721 cells via Wnt signaling pathway.
Fig. 7. Migration inhibition induced by compound 8b. (A) Anti-migration effect of compound 8b
on SMMC-7721 cells at concentrations of 0, 0.2 and 0.5 µM via transwell migration assay; (B)
Quantitative analysis of migration rate; (C) Inhibition of Wnt signaling pathway; (D) Staticstical
analysis of Wnt pathway protein expression levels. The results shown were representative of three
independent experiments. **: p < 0.01 verse control.
3
. Conclusion
Following our previous work, two series of novel Jiyuan Oridonin A
-1,2,3-triazole-azole hybrids were designed, synthesized and evaluated for their
anticancer activity against four selected human cancer cell lines. Most of the
synthesized compounds exhibited moderate to good activity against the tested cancer
cells and might serve as bioactive fragments and lead compounds for developing more
potent cytotoxic agents. Among them, compound 8b with an ethylidene substituted
thiazolidinedione moiety presented the best activity against Eca-109, MGC-803,
MCF-7, PC-3, A549 and SMMC-7721 cancer cell lines, with the IC50 values of
1
4

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2
.7±0.2 µM, 0.6±0.1 µM, 5.0±0.8 µM, 1.5±0.2 µM, 3.5±0.7 µM and 0.2±0.0 µM,
respectively. In addition, compound 8b showed good selectivity between cancer and
normal cells (70.2-fold selectivity to SMMC-7721 cells over HL-7702 cells). Further
mechanism studies revealed that compound 8b could obviously inhibit the
proliferation of SMMC-7721 cancer cells by inducing apoptosis and arresting the cell
cycle at G1 phase. Importantly, we also explored the effect on migration, and 8b
inhibited migration of SMMC-7721 cells in a concentration-dependent manner with
Wnt signaling pathway involved. More mechanism studies are underway and will be
reported in due course.
4
. Experimental
4
.1. General
All commercially available starting materials and solvents were reagent grade
and used without further purification. Column chromatography was carried out on
00−300 mesh silica gel (Qingdao Haiyang Chemical, China). Reactions were
2
monitored by thin-layer chromatography (TLC) on 0.25mm silicagel plates (GF254)
and visualized under UV light. Infrared (IR) spectra (KBr) were recorded on PE-1710
type instrument of PE Company (KBr pellet). Melting points were determined on an
1
13
X-5 micro-melting apparatus and are uncorrected. H NMR and C NMR spectra
were recorded on a Bruker 400 MHz and 101 MHz spectrometer, respectively. High
resolution mass spectra (HRMS) of all derivatives were recorded on a Waters
Micro-mass Q-T of Micro-mass spectrometer by electrospray ionization (ESI).
4
.2. General procedure for the synthesis of compound 2 and compound 3.
Jiyuan Oridonin A (150mg, 0.43mmol) was dissolved in THF (8mL),
3
-Bromo-1-propanol (150µL) and p-TsOH (10mg) were added to stir for 3h at room
temperature. Then the mixture was treated by heric-vacuum distillation and diluted
with water and extracted with ethyl acetate. The extract was washed with brine, dried
over anhydrous Na SO , filtered, evaporated and the crude product was purified by
2
4
column chromatography to give compound 2 (130mg, 75%). After that, compound 2
was put in the system of EtOAc/H O (5:1) with NaN reflux 7h at 80℃ to obtain
2
3
1
5

ACCEPTED MANUSCRIPT
compound 3.
.2.1. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-12-(3-bromopropoxy)-11,14-dihydr
4
oxy-4,4-dimethyl-8-methylenedecahydro-1H-6,11b-(epoxymethano)-6a,9-methanoc
yclohepta[a]naphthalen-7(8H)-one (Compound 2).
-1
White solid, 76% yield; Mp: 119-121 ℃ ; IR (KBr, cm ): 3608, 3362, 2951, 2869,
1
1
5
4
723, 1643, 1431, 1362, 1258, 1103, 1037, 654; H NMR (400 MHz, DMSO-d ) δ
6
.79, 5.38 (each 1H, s, H-17), 5.11 (1H, s, H-20), 5.10 (1H, d, J = 2.9 Hz, H-14-OH),
.51 (1H, s, H-11-OH), 4.45 (1H, d, J = 8.0 Hz, H-11α), 4.26 (1H, d, J = 7.0 Hz,
H-7β), 3.98-3.81 (2H, m, H-21), 2.80 (1H ,d, J = 8.9 Hz, H-13α), 2.68 (1H, d, J =
1
2.8 Hz, H-6β), 2.64 (1H, dd, J = 6.7 Hz, H-12α), 1.99 (1H, d, J = 12.4 Hz, H-1α),
.86-1.72 (2H, m, H-22), 1.61 (1H, d, J = 4.7 Hz, H-6α), 1.49-1.32 (3H, m, H-3α,
1
H-2), 1.29-1.26 (2H, m, H-1β, H-12β), 1.25-1.15 (2H, m, H-5β, H-9β), 1.11-1.05 (1H,
1
3
m, H-3β), 0.92 (3H, s, H-19), 0.82 (3H, s, H-18). C NMR (101 MHz, DMSO-d ) δ
6
2
3
06.20, 152.73, 119.69, 116.61, 99.94, 70.15, 65.85, 63.56, 62.88, 56.84, 48.90,
9.80, 39.59, 39.38, 34.17, 33.14, 30.31, 24.71, 21.19, 18.89, 18.23. ESI-HRMS: m/z
+
calcd. For C H BrNaO [M+Na] : 491.1409, found 491.1403.
2
3
33
5,
4
.2.2. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-12-(3-azidopropoxy)-11,14-dihydro
xy-4,4-dimethyl-8-methylenedecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocy
clohepta[a]naphthalen-7(8H)-one (Compound 3).
-1
White solid, 92% yield; Mp: 127-129 ℃ ; IR (KBr, cm ): 3648, 3612, 3421, 3136,
1
2
950, 2868, 1723, 1646, 1362, 1252, 1108, 1078, 1023, 766; H NMR (400 MHz,
DMSO-d ) δ 5.80, 5.38 (each 1H, s, H-17), 5.11 (1H, d, J = 2.9 Hz, H-14-OH), 5.10
6
(
1H, s, H-20), 4.51 (1H, s, H-11-OH), 4.45 (1H, d, J = 7.8 Hz, H-11α), 4.26 (1H, d, J
7.2Hz, H-7β), 3.99-3.82 (2H, m, H-21), 2.81 (1H, d, J = 9.1 Hz, H-13α), 2.68 (1H,
d, J = 12.9 Hz, H-6β), 2.60 (1H, s, H-12α), 2.00 (1H, d, J = 11.9 Hz, H-1α), 1.87-1.74
2H, m, H-22), 1.63-1.44 (1H, m, H-6α), 1.39 (2H, dd, J = 29.4, 11.2 Hz, H-2),
.36-1.27 (2H, m, H-1β, H-12β), 1.25-1.14 (2H, m, H-5β, H-9β), 1.09-1.01 (1H, m,
=
(
1
1
3
H-3β), 0.92 (3H, s, H-19), 0.84 (3H, s, H-18). C NMR (101 MHz, DMSO-d ) δ
6
2
06.25, 152.77, 116.54, 99.99, 70.13, 66.53, 65.78, 63.59, 57.77, 56.82, 51.24, 48.97,
1
6

ACCEPTED MANUSCRIPT
3
9.99, 39.78, 39.57, 39.36, 34.17, 33.17, 30.38, 24.75, 21.20, 18.25. ESI-HRMS: m/z
+
calcd. For C H N NaO [M+Na] : 454.2258, found 454.2256.
2
3
33
3
5,
4
.3. General procedure for the synthesis of the 4a-4f.
To a stirred solution of mercaptan (3.25 mmol, 1.25eq.) in acetone (15 mL),
propargyl bromide (3.00 mmol, 1.00eq) and K CO (3.05 mmol,1.02eq) were added
2
3
carefully and the reaction mixture was refluxed for 5h. Upon completion, the reaction
mixture was concentrated under vacuum, the residue was dissolved in EtOAc (30 mL)
and washed with water, brine, dried over anhydrous Na SO and concentrated under
2
4
vacuum to afford 4a-4f in 70%-86%, which were used in the next reaction without
further purification. The structure confirmation of 4a-4f were described in the
Supplementary data.
4
.4. General procedure to synthesize compounds 5a-5f.
Compound 3 (150mg, 2eq) was dissolved in t-BuOH (4mL) and appropriate alkynes
were dropped then put 1mL H O in it, finally, the 57mg(1eq) CuSO and 107mg(3eq)
2
4
Cu as catalytic agent to react 8.5h. Then insoluble matter was filtered and condensed,
subsequently, extracted with ethyl acetate and washed with brine, dried over
anhydrous Na SO , filtered, evaporated and the crude product was purified by column
2
4
chromatography to give compounds.
.4.1. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-dimethyl-12-(3
(4-(((1-methyl-1H-tetrazol-5-yl)thio)methyl)-1H-1,2,3-triazol-1-yl)propoxy)-8-meth
ylenedecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclohepta[a]naphthalen-
(8H)-one.(5a)
4
-
7
-1
White solid, 82% yield; Mp: 143-145℃ . IR (KBr, cm ) 3648, 3612, 3421, 3136, 2950,
1
2
868, 1723, 1646, 1362, 1268, 1145, 1018, 939, 669. H NMR (400 MHz, DMSO-d )
6
δ 8.05 (s, 1H, H-24), 5.80, 5.30 (each 1H, s, H-17), 5.09 (s, 2H, H-26), 4.59 (s, 2H,
H-20, H-14-OH), 4.49 (d, J = 26.2 Hz, 2H, H-11-OH, H-11α), 4.42 (t, 2H, H-21),
4
.29 (d, J = 2.9 Hz, 1H, H-7β), 3.99 (s, 1H, H-14α), 3.91 (s, 3H, H-28), 3.75 (s, 1H,
H-6β), 3.23 (s, 1H, H-13α), 2.77 (d, J = 29.7 Hz, 1H, H-12β), 2.66 (t, 2H, H-23), 2.51
d, J = 22.4 Hz, 1H, H-1α), 2.05 (m, 2H, H-22), 1.58 (s, 1H, H-6α), 1.39 (m, 1H,
(
1
7

ACCEPTED MANUSCRIPT
H-12α), 1.35 (m, 2H, H-1β, H-3α), 1.32 – 1.25 (m, 2H, H-2), 1.19 (m, 2H, H-5β,
1
3
H-9β), 1.08 (s, 1H, H-3β), 0.92 (s, 3H, H-19), 0.83 (s, 3H, H-18). C NMR (101 MHz,
DMSO-d ) δ 206.23, 153.45, 152.78, 142.61, 124.22, 116.57, 99.96, 70.09, 65.67,
6
6
3
4.69, 63.70, 60.14, 57.78, 56.68, 48.85, 47.22, 43.03, 42.35, 40.87, 34.17, 34.08,
3.16, 30.75, 30.58, 28.07, 24.76, 21.21, 18.24. HR-MS(ESI): Calculated for
+
C H N O SNa [M+Na] : 608.2631, found 608.2632.
28
39
7
5
4
.4.2. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-12-(3-(4-(((1,3,4-thiadiazol-2-yl)thi
o)methyl)-1H-1,2,3-triazol-1-yl)propoxy)-11,14-dihydroxy-4,4-dimethyl-8-methylene
decahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclohepta[a]naphthalen-7(8H)
-one.(5b)
-1
White solid, 76% yield; Mp: 163-165℃ . IR (KBr, cm ) 3648, 3612, 3430, 2928, 1752,
1
1
686, 1426, 1386, 1268, 1145, 1018, 938, 676. H NMR (400 MHz, DMSO-d ) δ 8.08
6
(s, 1H, H-24), 5.80, 5.39 (each 1H, s, H-17), 5.09 (s, 2H, H-26), 4.65 (s, 1H, H-28),
4.60 (s, 1H, H-20), 4.52 (s, 1H, H-14-OH), 4.47 (d, J = 24.4 Hz, 3H, H-21, H-11-OH),
4.28 (d, J = 5.9 Hz, 1H, H-11α), 3.99 (m, 1H, H-14α), 3.75 (d, J = 18.9 Hz, 1H, H-6β),
3.24 (d, J = 13.5 Hz, 1H, H-13α), 2.82 (d, J = 8.0 Hz, 1H, H-12β), 2.65 (d, J = 10.7
Hz, 2H, H-23), 2.11 (d, J = 12.6 Hz, 2H, H-22), 2.02 (d, J = 10.4 Hz, 1H, H-1α), 1.57
d, J = 12.1 Hz, 1H, H-6α), 1.48 – 1.40 (m, 2H, H-7β, H-12α), 1.35 (d, J = 24.6 Hz,
(
2
1
H, H-2), 1.31 – 1.23 (m, 2H, H-1β, H-3α), 1.20 (d, J = 6.5 Hz, 1H, H-5β), 1.15 (m,
1
3
H, H-9β), 1.09 (d, J = 10.6 Hz, 1H, H-3β), 0.91 (s, 3H, H-19), 0.82 (s, 3H, H-18). C
NMR (101 MHz, DMSO-d ) δ 206.22, 165.21, 154.79, 152.79, 142.69, 124.30,
6
116.56, 99.98, 70.09, 65.67, 64.71, 63.70, 57.78, 56.67, 48.85, 47.24, 43.03, 42.35,
4
0.87, 34.17, 33.16, 30.74, 30.58, 29.00, 28.88, 24.76, 21.21, 18.25. HR-MS(ESI):
+
Calculated for C H N O S Na [M+Na] : 610.2134, found 610.2139.
2
8
37
5
5 2
4
.4.3. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-dimethyl-8-me
thylene-12-(3-(4-((pyrimidin-2-ylthio)methyl)-1H-1,2,3-triazol-1-yl)propoxy)decahy
dro-1H-6,11b-(epoxymethano)-6a,9-methanocyclohepta[a]naphthalen-7(8H)-one.(5
c)
-1
White solid, 79% yield; Mp: 165-157℃ . IR (KBr, cm ) 3630, 3328, 1752, 1720, 1686,
1
8

ACCEPTED MANUSCRIPT
1
1
426, 1386, 1268, 1135, 1018, 939, 679. H NMR (400 MHz, DMSO-d ) δ 8.68 (d, J
6
=
4.8 Hz, 2H, H-28, H-30), 8.03 (s, 1H, H-24), 7.26 (t, J = 4.8 Hz, 1H, H-29), 5.81,
.39 (each 1H, s, H-17), 5.10 (d, J = 2.2 Hz, 1H, H-20), 5.08 (s, 1H, H-14-OH), 4.52
s, 1H, H-11-OH), 4.51 – 4.46 (m, 1H, H-11α), 4.45 (s, 2H, H-26), 4.41 (t, J = 7.0 Hz,
H, H-21), 4.27 (t, J = 8.9 Hz, 1H, H-7β), 3.98 (d, J = 2.7 Hz, 1H, H-14α), 3.74 (dd, J
10.0, 5.2 Hz, 1H, H-6β), 3.28 – 3.18 (m, 1H, H-13α), 2.81 (d, J = 8.9 Hz, 1H,
H-12β), 2.64 (dt, J = 17.3, 10.6 Hz, 2H, H-23), 2.10 – 2.04 (m, 2H, H-22), 2.00 (d, J =
.9 Hz, 1H, H-1α), 1.60 – 1.53 (m, 1H, H-6α), 1.45 – 1.39 (m, 2H, H-2), 1.35 – 1.28
m, 2H, H-1β, H-12α), 1.24 (d, J = 4.9 Hz, 1H, H-3α), 1.20 (d, J = 7.5 Hz, 1H, H-5β),
.18 (m, 1H, H-9β), 1.08 (d, J = 11.5 Hz, 1H, H-3β), 0.91 (s, 3H, H-19), 0.82 (s, 3H,
5
(
2
=
6
(
1
1
3
H-18). C NMR (101 MHz, DMSO-d ) δ 206.23, 170.71, 158.38, 152.79, 143.82,
6
1
4
1
6
4
23.85, 117.91, 116.57, 99.97, 70.08, 67.49, 65.66, 64.74, 63.68, 57.77, 56.66, 48.84,
7.13, 43.03, 42.36, 34.86, 34.17, 33.16, 30.88, 30.73, 30.57, 25.46, 24.75, 21.21,
+
8.24. HR-MS(ESI): Calculated for C H N O SNa [M+Na] : 604.2570, found
3
0
39
5
5
04.2572.
.4.4. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-dimethyl-12-
(
3-(4-(((5-methyl-1,3,4-thiadiazol-2-yl)thio)methyl)-1H-1,2,3-triazol-1-yl)propoxy)-8
methylenedecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclohepta[a]naphth
-
alen-7(8H)-one.(5d)
-1
White solid, 81% yield; Mp: 178-179℃. IR (KBr, cm ) 3630, 3430, 3328, 1752, 1720,
1
1
686, 1426, 1386, 1268, 1135, 1018, 939, 699. H NMR (400 MHz, DMSO-d ) δ 8.08
6
(s, 1H, H-24), 5.81, 5.34 (each 1H, s, H-17), 5.09 (s, 2H, H-26), 4.59 (d, J = 7.8 Hz,
2H, H-20, H-14-OH), 4.52 (s, 1H, H-11-OH), 4.50 – 4.46 (m, 1H, H-11α), 4.44 (t, J =
6.9 Hz, 2H, H-21), 4.28 (d, J = 7.0 Hz, 1H, H-7β), 3.99 (d, J = 3.0 Hz, 1H, H-14α),
3.81 – 3.70 (m, 1H, H-6β), 3.30 – 3.17 (m, 1H, H-13α), 2.82 (d, J = 9.0 Hz, 1H,
H-12β), 2.64 (dt, J = 13.8, 11.2 Hz, 2H, H-23), 2.11 (s, 3H, H-29), 2.03 (d, J = 11.4
Hz, 1H, H-1α), 1.62 – 1.52 (m, 1H, H-6α), 1.40 (dd, J = 23.7, 12.2 Hz, 4H, H-22,
H-1β, H-12α), 1.34 – 1.23 (m, 2H, H-2), 1.19 (t, J = 7.5 Hz, 2H, H-3α, H-5β), 1.13 (d,
J = 10.8 Hz, 1H, H-9β), 1.09 (d, J = 11.5 Hz, 1H, H-3β), 0.91 (s, 3H, H-19), 0.82 (s,
1
9

ACCEPTED MANUSCRIPT
1
3
3
1
4
H, H-18). C NMR (101 MHz, DMSO-d ) δ 206.22, 165.16, 152.78, 142.63, 124.30,
6
16.57, 99.98, 70.09, 65.67, 64.72, 63.70, 57.78, 56.67, 56.29, 48.85, 47.26, 43.03,
2.35, 40.87, 34.17, 33.16, 31.17, 30.74, 30.58, 30.07, 28.99, 24.76, 21.21, 18.25.
+
HR-MS(ESI): Calculated for C H N O S Na [M+Na] : 624.2290, found 624.2296.
2
9
39
5
5 2
4
.4.5. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-12-(3-(4-(((4,5-dihydrothiazol-2-yl)
thio)methyl)-1H-1,2,3-triazol-1-yl)propoxy)-11,14-dihydroxy-4,4-dimethyl-8-methyl
enedecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclohepta[a]naphthalen-7
(8H)-one.(5e)
-1
White solid, 77% yield; Mp: 172-174℃. IR (KBr, cm ) 3430, 2928, 1752, 1720, 1686,
1
1
1
4
426, 1386, 1268, 1145, 1018, 939, 669. H NMR (400 MHz, DMSO-d ) δ 7.99 (s,
6
H, H-24), 5.81, 5.37 (each 1H, s, H-17), 5.10 (s, 2H,H-26), 4.53 (s, 1H, H-14-OH),
.51 – 4.46 (m, 1H, H-20), 4.44 (d, J = 5.2 Hz, 1H, H-11α), 4.42 (d, J = 6.3 Hz, 3H,
H-21, H-11-OH), 4.29 (d, J = 7.0 Hz, 1H, H-7β), 4.18 (t, J = 8.0 Hz, 2H, H-13α,
H-12β), 3.99 (d, J = 3.1 Hz, 1H, H-14α), 3.81 – 3.71 (m, 1H, H-6β), 3.47 (t, J = 8.0
Hz, 2H, H-28), 3.25 (dt, J = 10.0, 6.4 Hz, 2H, H-29), 2.72 – 2.59 (m, 2H, H-23), 2.14
–
2.06 (m, 2H, H-22), 2.01 (d, J = 8.2 Hz, 1H, H-1α), 1.63 – 1.52 (m, 1H, H-6α), 1.45
d, J = 7.6 Hz, 1H, H-12α), 1.41 (d, J = 6.0 Hz, 1H, H-1β), 1.40 – 1.33 (m, 2H, H-2),
.29 (dd, J = 14.0, 8.4 Hz, 1H, H-3α), 1.19 (dd, J = 7.9, 4.5 Hz, 2H, H-5β, H-9β), 1.08
(
1
13
(
m, J = 11.7 Hz, 1H, H-3β), 0.92 (s, 3H, H-19), 0.83 (s, 3H, H-18). C NMR (101
MHz, DMSO-d ) δ 206.22, 163.01, 152.79, 143.17, 124.08, 116.56, 99.99, 70.09,
6
6
3
5.68, 64.75, 64.43, 63.70, 60.24, 57.78, 56.66, 48.84, 47.18, 43.04, 42.36, 40.87,
5.84, 34.18, 33.16, 30.75, 30.59, 27.09, 24.77, 21.22, 18.27. HR-MS(ESI):
+
Calculated for C H N O S Na [M+Na] : 611.2338, found 611.2336.
2
9
40
4
5 2
4
.4.6. (4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-12-(3-(4-((benzo[d]thiazol-2-ylthio)
methyl)-1H-1,2,3-triazol-1-yl)propoxy)-11,14-dihydroxy-4,4-dimethyl-8-methylened
ecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclohepta[a]naphthalen-7(8H)-
one.(5f)
-1
White solid, 83% yield; Mp: 182-184℃. IR (KBr, cm ) 3420, 3328, 2952, 1720, 1686,
1
1
426, 1386, 1268, 1145, 1018, 939, 658. H NMR (400 MHz, DMSO-d ) δ 8.12 (s, 1H,
6
2
0

ACCEPTED MANUSCRIPT
H-24), 8.02 (d, J = 7.7 Hz, 1H, H-29), 7.91 (d, J = 7.8 Hz, 1H, H-30), 7.49 (t, J = 7.3
Hz, 1H, H-31), 7.38 (t, J = 7.2 Hz, 1H, H-32), 5.81, 5.38 (each 1H, s, H-17), 5.08 (d,
J = 9.0 Hz, 2H, H-20, H-14-OH), 4.72 (s, 2H, H-26), 4.53 (s, 1H, H-11-OH), 4.51 –
4
.46 (m, 1H, H-11α), 4.44 (d, J = 6.7 Hz, 2H, H-21), 4.28 (d, J = 6.5 Hz, 1H, H-7β),
.99 (s, 1H, H-14α), 3.83 – 3.65 (m, 1H, H-6β), 3.24 (d, J = 8.4 Hz, 1H, H-13α), 2.81
3
(
d, J = 8.7 Hz, 1H, H-12β), 2.75 – 2.54 (m, 2H, H-23), 2.16 – 2.02 (m, 3H, H-1α,
H-22), 1.63 – 1.50 (m, 1H, H-6α), 1.39 (d, J = 9.5 Hz, 1H, H-12α), 1.38 (m, 2H, H-2),
.31 (d, J = 8.4 Hz, 1H, H-1β), 1.26 (dd, J = 17.3, 8.8 Hz, 1H, H-3α), 1.19 (d, J = 7.9
Hz, 2H, H-5β, H-9β), 1.07 (d, J = 9.8 Hz, 1H, H-3β), 0.88 (s, 3H, H-19), 0.81 (s, 3H,
1
1
3
H-18). C NMR (101 MHz, DMSO-d ) δ 206.21, 166.27, 153.07, 152.78, 142.87,
6
1
6
3
35.19, 126.85, 125.02, 124.29, 122.30, 121.73, 116.55, 99.98, 70.09, 65.68, 64.72,
3.70, 57.78, 56.67, 48.84, 47.23, 43.04, 42.35, 40.84, 34.14, 33.15, 31.16, 30.73,
0.57, 27.94, 24.75, 21.17, 18.24. HR-MS(ESI): Calculated for C H N O S Na
3
3
40
4
5 2
+
[
M+Na] : 659.2338, found 659.2335.
4
.5. 3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-dimet
hyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocycloh
epta[a]naphthalen-12-yl)ethoxy)propyl)-1H-1,2,3-triazol-4-yl)methyl)thiazolidine-2,
4
-dione. (6a)
-1
White solid, 87% yield; Mp: 152-153℃ . IR (KBr, cm ) 3430, 2928, 1752, 1720, 1686,
1
1
426, 1386, 1268, 1145, 1018, 939, 669. H NMR (400 MHz, DMSO-d
6
) δ 8.02 (s, 1H,
H-24), 5.80, 5.30 (each 1H, s, H-17), 5.10 (s, 1H, H-20), 5.08 (d, J = 2.8 Hz, 1H,
H-14-OH), 4.74 (dd, 2H, J = 10.4, 6.1 Hz, H-21), 4.52 (s, 1H, H-11-OH), 4.51 – 4.45
(
m, 1H, H-11α), 4.42 (t, J = 7.0 Hz, 2H, H-26), 4.36 (t, J = 5.1 Hz, 1H, H-7β), 4.26 (d,
J = 9.5 Hz, 2H, H-27), 3.99 (d, J = 3.1 Hz, 1H, H-14α), 3.81 – 3.71 (m, 1H, H-6β),
.45 (dt, J = 7.0, 1.8 Hz, 1H, H-12α), 3.26 (dd, J = 6.5, 3.6 Hz, 1H, H-13α), 2.82 (d, J
9.1 Hz, 1H, H-12β), 2.71 – 2.58 (m, 2H, H-23), 2.07 (dd, J = 12.7, 6.1 Hz, 2H,
3
=
H-22), 2.01 (d, J = 9.1 Hz, 1H, H-1α), 1.62 – 1.53 (m, 1H, H-6α), 1.37 – 1.27 (m, 2H,
H-2), 1.27 – 1.23 (m, 2H, H-1β, H-3α), 1.22 – 1.20 (m, 1H, H-5β), 1.19 – 1.16 (m, 1H,
13
H-9β), 1.13 – 1.07 (m, 1H, H-3β), 0.93 (s, 3H, H-19), 0.83 (s, 3H, H-18). C NMR
2
1

ACCEPTED MANUSCRIPT
(101 MHz, DMSO-d
6
) δ 206.21, 172.38, 171.95, 152.80, 141.69, 124.07, 116.53,
9
4
9.99, 70.09, 65.68, 64.79, 63.70, 57.79, 56.70, 56.49, 48.86, 47.21, 43.04, 42.36,
0.89, 39.92, 36.80, 34.44, 34.17, 33.17, 30.70, 30.57, 24.76, 21.22, 18.24.
+
HR-MS(ESI): Calculated for C H N O SNa [M+Na] : 609.2359, found 609.2352.
2
9
38
4
7
4
.6. General procedure for the synthesis of compounds 7a-7k.
Equimolar quantities of substituted appropriate aldehydes 200mg and
thiazolidinedione 150mg in absolute methanol with few drops of glacial acetic acid
and piperidine(0.05eq) were refluxed for 2-3 h. The solvent was removed under
reduced pressure; residue left behind was poured in crushed ice, filtered, washed with
ice cold water, dried and the crude product was purified by column chromatography
to yield the final derivatives. The purified compounds were subsequently
1
13
characterized using H NMR, C NMR. The Z or E configuration of 7a-7k was
determined by the foresight method to calculate the chemical shifts of the protons on
variously substituted olefins [46]. Sohda,T have reported the calculated value of the
1
methine protons of Z products in H NMR spectra to be 7.80 ppm, which is consistent
with our tested value, and the value of E-products to be 7.14 ppm [47]. The NMR data
of compounds (7a-7k) were listed in supplementary material.
4
.7. General procedure for the synthesis of compounds 8a-8k.
Compound 3 (150mg, 2.00eq) was dissolved in t-BuOH (6mL) and compound 7a-7k
120mg, 1.05eq) were dropped then put 3mL H O in the system. Finally, the mixture
(
2
was stirred at room temperature for 6-12 h by using CuSO (53 mg, 1.00eq) and Cu
4
(103 mg, 3.00eq) as catalytic agents and then was filtered and condensed.
Subsequently, the residue was extracted with ethyl acetate, the organic layer was
washed with brine, dried over anhydrous Na SO , filtered, evaporated and the crude
2
4
product was purified by column chromatography to give compounds.
.7.1. (E)-3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-
4
dimethyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)-6a,9-methanoc
yclohepta[a]naphthalen-12-yl)ethoxy)propyl)-1H-1,2,3-triazol-4-yl)methyl)-5-ethyli
denethiazolidine-2,4-dione.(8a)
2
2

ACCEPTED MANUSCRIPT
-1
White solid, 84% yield; Mp: 158-159 ℃ . IR（KBr, cm ）: 3428, 2931, 2870, 1724,
1
1
689, 1636, 1426, 1365, 1336, 1264, 1158, 1101, 1018, 792. H NMR (400 MHz,
DMSO-d
H-17), 5.13 – 5.04 (m, 2H, H-20, H-14-OH), 4.86 (d, J = 15.5 Hz, 2H, H-21), 4.52 (s,
H, H-11-OH), 4.50 – 4.45 (m, 1H, H-11α), 4.42 (t, J = 7.0 Hz, 2H, H-26), 4.27 (d, J
7.0 Hz, 1H, H-7β), 3.98 (d, J = 3.1 Hz, 1H, H-14α), 3.75 (dt, J = 10.6, 5.5 Hz, 1H,
6
) δ 8.06 (s, 1H, H-24), 7.08 (t, J = 7.6 Hz, 1H, H-30), 5.80, 5.30 (each 1H, s,
1
=
H-6β), 3.23 (dt, J = 9.9, 6.4 Hz, 1H, H-13α), 2.83 (t, J = 9.8 Hz, 1H, H-12α), 2.64 (dt,
J = 14.2, 11.0 Hz, 2H, H-23), 2.24 (p, J = 7.5 Hz, 2H, H-22), 2.08 (dt, J = 13.4, 6.7
Hz, 3H, H-31), 2.04 (s, 1H, H-1α), 1.62 – 1.51 (m, 1H, H-6α), 1.45 (s, 1H, H-2α),
1
.41 (s, 1H, H-12β), 1.38 (s, 1H, H-2β), 1.32 – 1.27 (m, 1H, H-1β), 1.25 (d, J = 4.6
Hz, 1H, H-3α), 1.20 (s, 1H, H-5β), 1.18 (s, 1H, H-9β), 1.09 (t, J = 7.5 Hz, 1H, H-3β),
1
3
0
1
6
3
.91 (s, 3H, H-19), 0.82 (s, 3H, H-18). C NMR (101 MHz, DMSO-d ) δ 206.23,
6
67.23, 164.44, 152.79, 141.58, 140.85, 124.24, 124.02, 116.54, 99.98, 70.07, 65.66,
4.74, 63.68, 60.23, 57.77, 56.67, 48.84, 47.19, 43.03, 42.35, 40.87, 39.90, 36.90,
4.16, 33.17, 30.65, 30.57, 25.26, 24.76, 21.20, 18.22, 12.55. HR-MS(ESI)
+
Calculated for C H N O SNa [M+Na] : 649.2672, found 649.2679.
3
2
42
4
7
4
.7.2. (E)-3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-
dimethyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)-6a,9-methanoc
yclohepta[a]naphthalen-12-yl)ethoxy)propyl)-1H-1,2,3-triazol-4-yl)methyl)-5-methy
lenethiazolidine-2,4-dione.(8b)
-1
White solid, 82% yield; Mp: 155-156℃ . IR（KBr, cm ）: 3425, 2953, 2869, 1724,
1
1
687, 1638, 1426, 1365, 1334, 1102, 1019, 724, 655, 597, 552. H NMR (400 MHz,
DMSO-d
H-17), 5.09 (s, 2H, H-20, H-14-OH), 4.84 (s, 2H, H-25), 4.53 (s, 1H, H-11-OH), 4.48
s, 1H, H-11α), 4.42 (d, 1H, H-26), 4.27 (d, J = 6.3 Hz, 1H, H-7β), 4.02 (d, J = 6.3 Hz,
6
) δ 8.05 (s, 1H, H-24), 7.14 (d, J = 7.1 Hz, 1H, H-30), 5.80, 5.30 (each 1H, s,
(
1H, H-14α), 3.76 (s, 1H, H-6β), 3.25 (s, 1H, H-13α), 2.82 (d, J = 8.6 Hz, 1H, H-12α),
2.65 (d, J = 11.9 Hz, 2H, H-21), 2.19 (s, 1H, H-1α), 2.08 (s, 2H, H-22), 1.94 (d, J =
6.9 Hz, 2H, H-23), 1.55 (s, 1H, H-6α), 1.41 (s, 1H, H-3α), 1.36 (s, 2H, H-2), 1.28 (s,
1H, H-1β), 1.24 (s, 1H, H-12β), 1.21 (s, 1H, H-5β), 1.19 (s, 1H, H-9β), 1.08 (s, 1H,
2
3

ACCEPTED MANUSCRIPT
1
3
H-3β), 0.92 (s, 3H, H-19), 0.83 (s, 3H, H-18). C NMR (101 MHz, DMSO-d
6
) δ
2
6
3
06.23, 167.20, 164.25, 152.80, 141.59, 135.41, 125.66, 124.00, 116.54, 99.99, 70.08,
7.49, 65.68, 64.76, 63.69, 57.78, 56.68, 48.85, 47.20, 43.04, 42.35, 40.88, 40.61,
6.88, 34.16, 33.17, 30.89, 30.65, 27.68, 24.76, 21.20, 18.23, 17.47. HR-MS (ESI)
+
Calculated for C H N O SNa [M+Na] : 635.2515, found 635.2511.
3
1
40
4
7
4
.7.3. 3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-dime
thyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclo
hepta[a]naphthalen-12-yl)ethoxy)propyl)-1H-1,2,3-triazol-4-yl)methyl)-5-(propan-2
-ylidene)thiazolidine-2,4-dione.(8c)
-1
White solid, 85% yield; Mp: 164-165℃ .I R（ KBr，cm ）3429, 2951, 2869, 1725, 1680,
1
1
8
2
624, 1425, 1364, 1335, 1103, 1050, 1019, 659. H NMR (400 MHz, DMSO-d
6
) δ
.09 (s, 1H, H-24), 5.81, 5.38 (each 1H, s, H-17), 5.09 (s, 2H, H-26), 4.86 – 4.77 (m,
H, H-20, H-14-OH), 4.53 (s, 1H, H-11-OH), 4.49 (d, J = 7.8 Hz, 1H, H-11α), 4.43
(dd, J = 13.9, 7.2 Hz, 2H, H-21), 4.27 (d, J = 7.0 Hz, 1H, H-7β), 3.99 (d, J = 3.0 Hz,
1
H, H-14α), 3.76 (dd, J = 10.1, 5.3 Hz, 1H, H-6β), 3.24 (dt, J = 10.0, 6.5 Hz, 1H,
H-13α), 2.82 (d, J = 9.0 Hz, 1H, H-12β), 2.73 – 2.58 (m, 2H, H-23), 2.41 (s, 2H,
H-22), 2.07 (dd, J = 12.6, 6.2 Hz, 3H, H-31), 2.00 (s, 3H, H-32), 1.77 (dd, J = 8.0, 4.8
Hz, 1H, H-1α), 1.57 (dd, J = 8.5, 5.7 Hz, 1H, H-6α), 1.45 (s, 1H, H-12α), 1.41 (s, 1H,
H-2α), 1.38 (s, 1H, H-1β), 1.32 – 1.27 (m, 1H, H-2β), 1.25 (d, J = 4.7 Hz, 1H, H-3α),
1
.20 (s, 1H, H-5β), 1.18 (s, 1H, H-9β), 1.09 (d, J = 10.7 Hz, 1H, H-3β), 0.92 (s, 3H,
1
3
H-19), 0.83 (s, 3H, H-18). C NMR (101 MHz, DMSO-d
6
) δ 206.23, 166.53, 164.07,
1
5
3
52.80, 151.79, 141.80, 123.96, 116.54, 99.99, 70.08, 67.49, 65.67, 64.75, 63.69,
7.77, 56.68, 48.85, 47.18, 43.04, 42.35, 40.87, 39.91, 36.59, 34.16, 33.17, 30.89,
0.66, 27.01, 25.60, 24.76, 22.18, 21.20, 18.24. HR-MS(ESI) Calculated for
+
C H N O SNa [M+Na] : 649.2672, found 649.2676.
32
42
4
7
4
.7.4. (E)-3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-
dimethyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)-6a,9-methanoc
2
4

ACCEPTED MANUSCRIPT
yclohepta[a]naphthalen-12-yl)ethoxy)propyl)-1H-1,2,3-triazol-4-yl)methyl)-5-hexyli
denethiazolidine-2,4-dione.(8d)
-1
White solid, 90% yield; Mp: 172-173℃ . IR（KBr, cm ）: 3426, 2954, 2930, 2869,
1
1
724, 1688, 1635, 1425, 1365, 1333, 1147, 1019, 794, 657. H NMR (400 MHz,
DMSO-d ) δ 8.06 (s, 1H, H-24), 7.09 (d, J = 7.3 Hz, 1H, H-30), 5.81, 5.39 (each 1H, s,
6
H-17), 5.09 (s, 2H, H-26), 4.86 – 4.77 (m, 2H, H-20, H-14-OH), 4.53 (s, 1H,
H-11-OH), 4.51 – 4.46 (m, 1H, H-11α), 4.43 (d, 2H, H-21), 4.27 (d, J = 6.5 Hz, 1H,
H-7β), 3.99 – 3.76 (m, 2H, H-21), 3.25 (s, 1H, H-13α), 2.82 (d, J = 8.6 Hz, 1H,
H-12β), 2.65 (d, J = 11.5 Hz, 2H, H-23), 2.23 (d, J = 7.1 Hz, 2H, H-22), 2.13 – 1.97
(m, 3H, H-1α, H-31), 1.57 (s, 1H, H-6α), 1.51 (m, 2H, H-2), 1.44 (d, J = 5.5 Hz, 1H,
H-1β), 1.38 (m, 3H, H-3α, H-32), 1.29 (m, 4H, H-34, H-33), 1.19 (d, J = 7.9 Hz, 2H,
H-5β, H-9β), 1.09 (d, J = 10.8 Hz, 1H, H-3β), 0.92 (s, 3H, H-19), 0.87 (d, 3H, H-35),
1
3
0
1
5
3
.83 (s, 3H, H-18). C NMR (101 MHz, DMSO-d ) δ 206.22, 167.23, 164.35, 152.80,
6
41.57, 139.62, 124.90, 124.05, 116.52, 99.99, 70.08, 65.67, 64.75, 63.69, 57.78,
6.68, 48.85, 47.20, 43.04, 42.35, 40.88, 39.99, 36.91, 34.16, 33.17, 31.72, 31.27,
0.64, 27.38, 24.77, 22.29, 21.20, 18.23, 14.28. HR-MS(ESI) Calculated for
+
C H N O SNa [M+Na] : 691.3141, found 691.3127.
35
48
4
7
4
.7.5. 5-((Z)-benzylidene)-3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14
-
dihydroxy-4,4-dimethyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)
6a,9-methanocyclohepta[a]naphthalen-12-yl)ethoxy)propyl)-1H-1,2,3-triazol-4-yl)
-
methyl)thiazolidine-2,4-dione.(8e)
-1
White solid, 80% yield; Mp: 184-185℃ . IR（KBr, cm ）: 3414, 2929, 2869, 1723,
1
1
656, 1624, 1546, 1449, 1392, 1365, 1300, 1219, 1101, 1018, 768, 669. H NMR
(400 MHz, DMSO-d
6
) δ 8.62 (t, J = 5.1 Hz, 1H, H-30), 7.95 (s, 1H, H-24), 7.56 (d, J
=
6.9 Hz, 1H, H-34), 7.44 (d, J = 9.0 Hz, 2H, H-32, H-33), 7.40 (d, J = 9.9 Hz, 2H,
H-35, H-36), 6.72 – 6.60 (d, 1H, H-20), 5.81, 5.37 (each 1H, s, H-17), 5.18 – 5.02 (m,
H, H-12α, H-14-OH), 4.52 (s, 1H, H-11-OH), 4.48 (d, J = 12.0 Hz, 2H, H-26), 4.46
t, 1H, H-11α), 4.36 (t, J = 5.2 Hz, 1H, H-7β), 4.28 (d, J = 7.0 Hz, 2H, H-21), 3.99 (d,
J = 2.6 Hz, 1H, H-14α), 3.82 – 3.71 (m, 1H, H-6β), 3.26 (dd, J = 14.9, 5.6 Hz, 1H,
2
(
2
5

ACCEPTED MANUSCRIPT
1
3α), 2.82 (d, J = 8.9 Hz, 1H, 12β), 2.72 – 2.58 (t, 2H, H-23), 2.08 (dd, J = 14.8, 8.6
Hz, 2H, H-22), 2.01 (d, J = 9.1 Hz, 1H, H-1α), 1.56 (dd, J = 8.8, 5.7 Hz, 1H, H-6α),
.42 (s, 1H, 12α), 1.39 (m, 2H, H-2), 1.32 – 1.23 (m, 2H, H-1β, H-3α), 1.20 (s, 1H,
H-5β), 1.18 (s, 1H, H-9β), 1.06 (d, J = 6.9 Hz, 1H, H-3β), 0.91 (s, 3H, H-19), 0.80 (s,
1
1
3
3
1
6
3
H, H-18). C NMR (101 MHz, DMSO-d ) δ 206.21, 172.38, 171.95, 152.80, 145.31,
6
41.69, 135.28, 128.69, 128.53, 128.06, 124.07, 116.53, 116.17, 99.99, 70.09, 65.68,
4.79, 63.70, 57.79, 56.70, 56.49, 48.86, 47.21, 43.04, 42.36, 40.89, 39.92, 36.80,
4.16, 33.17, 30.71, 30.57, 24.76, 21.21, 18.23. HR-MS(ESI): Calculated for
+
C H N O SNa [M+Na] : 697.2672, found 697.2671.
36
42
4
7
4
.7.6. 5-((Z)-3,4-difluorobenzylidene)-3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12
S,14R)-11,14-dihydroxy-4,4-dimethyl-8-methylene-7-oxododecahydro-1H-6,11b-(ep
oxymethano)-6a,9-methanocyclohepta[a]naphthalen-12-yl)oxy)propyl)-1H-1,2,3-tri
azol-4-yl)methyl)thiazolidine-2,4-dione.(8f)
-1
White solid, 89% yield; Mp: 197-198℃ . IR（KBr, cm ）: 3423, 2953, 2870, 1722,
1
1
664, 1629, 1517, 1435, 1365, 1277, 1113, 1018, 819, 663, 584. H NMR (400 MHz,
DMSO-d ) δ 8.63 (t, J = 5.6 Hz, 1H, H-30), 7.95 (s, 1H, H-24), 7.71 – 7.63 (m, 1H,
6
H-32), 7.50 (dd, J = 13.6, 5.3 Hz, 1H, H-35), 7.46 (d, J = 6.3 Hz, 1H, H-36), 6.65 (d,
J = 15.8 Hz, 1H, H-14-OH), 5.81, 5.37 (each 1H, s, H-17), 5.09 (d, J = 3.0 Hz, 2H,
H-26), 4.52 (s, 1H, H-11-OH), 4.48 (d, J = 9.3 Hz, 1H, H-20), 4.45 (d, J = 6.1 Hz, 2H,
H-21), 4.42 (s, 1H, H-11α), 4.28 (d, J = 7.1 Hz, 1H, H-7β), 3.99 (d, J = 3.1 Hz, 1H,
H-14α), 3.81 – 3.73 (m, 1H, H-6β), 3.26 (dt, J = 9.9, 6.4 Hz, 1H, H-13α), 2.82 (d, J =
9
.0 Hz, 1H, 12β), 2.72 – 2.58 (m, 2H, H-23), 2.09 (dt, J = 13.4, 6.7 Hz, 2H, H-22),
.00 (d, J = 7.3 Hz, 1H, H-1α), 1.62 – 1.51 (m, 1H, H-6α), 1.42 (s, 1H, H-12α), 1.39
2
(s, 2H, H-2), 1.32 – 1.27 (m, 1H, H-1β), 1.25 (d, J = 8.7 Hz, 1H, H-3α), 1.21 (s, 1H,
H-5β), 1.18 (s, 1H, H-9β), 1.06 (t, J = 7.0 Hz, 1H, H-3β), 0.91 (s, 3H, H-19), 0.80 (s,
1
3
3
1
9
H, H-18). C NMR (101 MHz, DMSO-d ) δ 206.21, 172.38, 171.95, 152.80, 151.38,
6
50.23, 145.31, 141.69, 132.59, 126.54, 125.19, 124.07, 123.40, 116.53, 116.17,
9.99, 70.09, 65.68, 64.79, 63.70, 57.79, 56.70, 56.49, 48.86, 47.21, 43.04, 42.36,
2
6

ACCEPTED MANUSCRIPT
4
0.89, 39.92, 36.80, 34.17, 33.17, 30.70, 30.57, 24.76, 21.22, 18.24. HR-MS(ESI):
+
Calculated for C H F N O SNa [M+Na] : 733.2483, found 733.2482.
3
6
40
2
4
7
4
.7.7. 5-((Z)-2-chlorobenzylidene)-3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14
R)-11,14-dihydroxy-4,4-dimethyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxy
methano)-6a,9-methanocyclohepta[a]naphthalen-12-yl)oxy)propyl)-1H-1,2,3-triazol
-4-yl)methyl)thiazolidine-2,4-dione.(8g)
-1
White solid, 86% yield; Mp: 181-182℃ . IR（KBr, cm ）：3413, 3069, 2949, 2869,
1
7
722, 1660, 1624, 1546, 1470, 1442, 1364, 1270, 1219, 1102, 1051, 1018, 975, 955,
59, 677, 594. 1H NMR (400 MHz, DMSO-d ) δ 8.75 (t, J = 5.6 Hz, 1H, H-30), 7.97
6
(s, 1H, H-24), 7.77 (d, J = 15.7 Hz, 1H, H-33), 7.70 (dd, J = 5.8, 3.7 Hz, 1H, H-34),
7.54 (dq, J = 7.4, 3.8 Hz, 1H, H-35), 7.46 – 7.38 (m, 1H, H-36), 6.72 (d, J = 15.7 Hz,
1H, H-14-OH), 5.80, 5.37 (each 1H, s, H-17), 5.09 (d, J = 3.1 Hz, 2H, H-26), 4.52 (s,
1H, H-20), 4.48 (d, J = 10.3 Hz, 1H, H-11-OH), 4.45 (d, J = 6.1 Hz, 2H, H-21), 4.35
(
d, J = 4.8 Hz, 1H, H-11α), 4.28 (d, J = 7.1 Hz, 1H, H-7β), 3.99 (d, J = 3.1 Hz, 1H,
H-14α), 3.82 – 3.73 (m, 1H, H-6β), 3.25 (dt, J = 10.0, 6.5 Hz, 1H, H-13α), 2.82 (d, J
9.0 Hz, 1H, H-12β), 2.72 – 2.59 (m, 2H, H-23), 2.09 (dt, J = 13.1, 6.7 Hz, 2H,
H-22), 2.01 (d, J = 9.3 Hz, 1H, H-1α), 1.61 – 1.50 (m, 1H, H-6α), 1.42 (s, 1H, H-12α),
=
1
.39 (s, 1H, H-1β), 1.34 – 1.27 (m, 2H, H-2), 1.25 (d, J = 8.7 Hz, 1H, H-3α), 1.20 (s,
H, H-5β), 1.18 (s, 1H, H-9β), 1.07 (t, J = 5.6 Hz, 1H, H-3β), 0.90 (s, 3H, H-19), 0.80
1
1
3
(
s, 3H, H-18). C NMR (101 MHz, DMSO-d ) δ 206.21, 172.38, 171.95, 152.80,
6
1
45.31, 141.69, 135.10, 133.09, 130.39, 130.17, 127.98, 126.79, 124.07, 116.53,
116.17, 99.99, 70.09, 65.68, 64.79, 63.70, 57.79, 56.70, 56.49, 48.86, 47.21, 43.04,
4
2.36, 40.89, 39.92, 36.80, 34.17, 33.17, 30.70, 30.57, 24.76, 21.22, 18.24.
+
HR-MS(ESI): Calculated for C H ClN O SNa [M+Na] : 731.2282, found 731.2281.
3
6
41
4
7
4
.7.8. 3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-dim
ethyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclo
hepta[a]naphthalen-12-yl)oxy)propyl)-1H-1,2,3-triazol-4-yl)methyl)-5-((Z)-4-nitrob
enzylidene)thiazolidine-2,4-dione.(8h)
2
7

ACCEPTED MANUSCRIPT
-1
White solid, 90% yield; Mp: 207-208℃ . IR（KBr, cm ）: 3413, 2925, 2869, 1721,
1
1
654, 1630, 1520, 1449, 1344, 1222, 1106, 1018, 864, 669. H NMR (400 MHz,
DMSO-d
.97 (s, 1H, H-24), 7.89 – 7.79 (m, 2H, H-35, H-36), 7.65 – 7.53 (d, 1H, H-33), 6.86
dd, J = 14.2, 4.4 Hz, 1H, H-14-OH), 5.80, 5.37 (each 1H, s, H-17), 5.09 (dd, J = 6.5,
.6 Hz, 2H, H-26), 4.52 (s, 1H, H-11-OH), 4.49 (d, J = 5.4 Hz, 1H, H-11α), 4.45 (d, J
8.6 Hz, 2H, H-21), 4.37 (dd, J = 5.4, 2.9 Hz, 1H, H-7β), 4.30 – 4.25 (m, 1H, H-20),
6
) δ 8.79 (dd, J = 12.4, 6.7 Hz, 1H, H-30), 8.27 (d, J = 8.8 Hz, 1H, H-32),
7
(
3
=
3.99 (d, J = 3.1 Hz, 1H, H-14α), 3.82 – 3.72 (m, 1H, H-6β), 3.26 (dt, J = 9.9, 6.4 Hz,
1H, H-13α), 2.82 (d, J = 9.0 Hz, 1H, H-12β), 2.72 – 2.58 (m, 2H, H-23), 2.09 (dt, J =
15.0, 7.4 Hz, 2H, H-22), 2.05 – 1.98 (m, 1H, H-1α), 1.61 – 1.51 (m, 1H, H-6α), 1.42
(s, 1H, H-12α), 1.39 (m, 2H, H-2), 1.30 – 1.23 (m, 2H, H-1β, H-3α), 1.19 (d, J = 9.8
Hz, 2H, H-5β, H-9β), 1.07 (t, J = 5.6 Hz, 1H, H-3β) 0.91 (s, 3H, H-19), 0.80 (s, 3H,
1
3
H-18). C NMR (101 MHz, DMSO-d
6
) δ 206.21, 172.38, 171.95, 152.80, 149.23,
1
6
3
43.38, 145.31, 141.69, 130.09, 124.37, 124.07, 116.53, 116.17, 99.99, 70.09, 65.68,
4.79, 63.70, 57.79, 56.70, 56.49, 48.86, 47.21, 43.04, 42.36, 40.89, 39.92, 36.80,
4.17, 33.17, 30.70, 30.57, 24.76, 21.22, 18.24. HR-MS(ESI): Calculated for
+
C H N O SNa [M+Na] : 742.2523, found 742.2521.
36
41
5
9
4
.7.9. 3-((1-(3-(((4aR,6R,6aS,9S,11S,11aS,11bS,12S,14R)-11,14-dihydroxy-4,4-dim
ethyl-8-methylene-7-oxododecahydro-1H-6,11b-(epoxymethano)-6a,9-methanocyclo
hepta[a]naphthalen-12-yl)ethoxy)propyl)-1H-1,2,3-triazol-4-yl)methyl)-5-((Z)-3-met
hoxybenzylidene)thiazolidine-2,4-dione.(8i)
-1
White solid, 80% yield; Mp: 214-215℃ . IR（KBr, cm ）：3412, 2929, 2868, 1720,
1
1
654, 1618, 1578, 1451, 1365, 1292, 1256, 1047, 889, 848, 790, 669, 558. H NMR
(400 MHz, DMSO-d
6
) δ 8.61 (t, J = 5.6 Hz, 1H, H-30), 7.97 (s, 1H, H-24), 7.44 (d, J
=
15.8 Hz, 1H, H-32), 7.33 (t, J = 7.9 Hz, 1H, H-34), 7.17 – 7.10 (m, 2H, H-35, H-36),
6
.96 (s, 1H, H-14-OH), 6.66 (s, 1H, H-20), 5.80, 5.37 (each 1H, s, H-17), 5.10 (d, J =
.8 Hz, 2H, H-26), 4.52 (s, 1H, H-11-OH), 4.48 (d, J = 8.4 Hz, 1H, H-11α), 4.43 (dd,
2
J = 9.2, 4.2 Hz, 4H, H-12α, H-37), 4.28 (dd, J = 12.3, 7.5 Hz, 1H, H-7β), 3.99 (t, J =
.1 Hz, 1H, H-14α), 3.78 (m, 2H, H-21), 3.77 – 3.74 (m, 1H, H-6β), 3.25 (dt, J = 10.0,
5
2
8