Design and synthesis of novel nitrogen mustard-evodiamine hybrids with selective antiproliferative activity

Article abstract of DOI:10.1016/j.bmcl.2017.10.014

A series of novel nitrogen mustard-evodiamine hybrids were synthesized and evaluated for their antitproliferative properties. The antiproliferative activities of 10a–d, 11a–d, and 12a–d against four different kinds of human cancer cell lines (PC-3, HepG2, THP-1 and HL-60) and human normal peripheral blood mononuclear cells (PBMC) were determined. The results showed that all the target hybrid compounds exhibited antiproliferative activities against tested human tumor cell lines to some extent and no antiproliferative activities (>200 μM) against human normal PBMC cells. The antiproliferative selectivity between tumorous and normal cells was very useful for further antitumor drug development. Among the target compounds, 12c showed the strongest cytotoxicity against two tumor cell lines (THP-1 and HL-60) with IC₅₀ values of 4.05 μM and 0.50 μM, respectively, and selected for further mechanism study in HL-60 cells. The results showed that 12c could induce HL-60 cells apoptosis and arrest at G₂ phase at low sub-micromolar concentrations via mitochondria-related pathways.

Full text of DOI:10.1016/j.bmcl.2017.10.014

Bioorganic & Medicinal Chemistry Letters xxx (2017) xxx–xxx
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Design and synthesis of novel nitrogen mustard-evodiamine hybrids
with selective antiproliferative activity
a
b
a
a
a
a,
⇑
c
c
Xu Hu , Yan Wang , Jingjing Xue , Tong Han , Runwei Jiao , Zhanlin Li , Weiwei Liu , Fanxing Xu ,
a,
⇑
a,
⇑
Huiming Hua , Dahong Li
a
Key Laboratory of Structure-Based Drug Design & Discovery, Ministry of Education, and School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University,
03 Wenhua Road, Shenyang 110016, PR China
Valiant Co. Ltd., 11 Wuzhishan Road, YEDA Yantai, Shandong 264006, PR China
1
b
c
Wuya College of Innovation, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, 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 nitrogen mustard-evodiamine hybrids were synthesized and evaluated for their antit-
proliferative properties. The antiproliferative activities of 10a–d, 11a–d, and 12a–d against four different
kinds of human cancer cell lines (PC-3, HepG2, THP-1 and HL-60) and human normal peripheral blood
mononuclear cells (PBMC) were determined. The results showed that all the target hybrid compounds
exhibited antiproliferative activities against tested human tumor cell lines to some extent and no
Received 23 August 2017
Revised 26 September 2017
Accepted 7 October 2017
Available online xxxx
antiproliferative activities (>200 lM) against human normal PBMC cells. The antiproliferative selectivity
between tumorous and normal cells was very useful for further antitumor drug development. Among the
target compounds, 12c showed the strongest cytotoxicity against two tumor cell lines (THP-1 and HL-60)
with IC50 values of 4.05 lM and 0.50 lM, respectively, and selected for further mechanism study in HL-60
Keywords:
Nitrogen mustard
Evodiamine
Hybrid
Antiproliferative activity
2
cells. The results showed that 12c could induce HL-60 cells apoptosis and arrest at G phase at low sub-
micromolar concentrations via mitochondria-related pathways.
Ó 2017 Published by Elsevier Ltd.
5
Alkaloids have long played important roles in cancer
chemotherapy. Evodiamine (1, Scheme 1) is a quinolone alkaloid
potential in human renal carcinoma cells and human ovarian can-
1
14
cer cells, suppress ABCG2 mediated drug resistance by inhibiting
p50/p65 NF-jB pathway in colorectal cancer and so on. Besides,
it could also sensitize chemoresistant breast cancer cells to adri-
amycin without obvious cytotoxicity against normal human
1
5
isolated from the fruits of traditional Chinese herb Evodiae fructus,
which is widely used for the treatment of diverse human disor-
ders.2 Previous studies showed that evodiamine possessed exten-
3
–5
16
sive biological activities, such as antitumor,
disease, anti-inflammatory and so on.
anti-Alzheimer’s
peripheral blood cells. Although evodiamine exhibited safe and
6
7,8
9,10
Recent studies have
known broad antitumor activity, the development of evodiamine
for cancer therapy was stunted by its relatively moderate potency.
Therefore, it is very necessary to develop novel evodiamine deriva-
tives with improved antiproliferative activity by structural modifi-
cation. To date, some evodiamine derivatives A–F
been designed and synthesized, which also rouse our interest for
further investigation.
Nitrogen mustards are DNA alkylating agents which have been
widely used in cancer chemotherapy with the advantages of broad
spectrum of antitumor activity and strong antiproliferative
reported that evodiamine exhibited cytotoxicity against a wide
variety of tumor cells in vitro and antitumor activities in vivo by
mainly inducing apoptosis or cell cycle arrest, and inhibiting topoi-
somerase I and II, angiogenesis, invasion, and metastasis.1 In addi-
tion, many researchers tried to elucidate the mechanisms of its
antitumor effects. For instance, evodiamine could block STAT3 sig-
naling pathway by inducing SHP-1 in hepatocellular carcinoma
1
17–22
(Fig. 1) have
1
2
cells, inhibit the activation of PI3K/Akt pathway, phosphorylation
of PTEN and mammalian target of rapamycin (mTOR), and the
activity of cAMP-dependent protein kinase A (PKA) in pancreatic
2
3
potency. However, the lack of selectivity causes serious side
effects, and acquired drug resistance limits the use of nitrogen
1
3
cancer, inhibit JNK- and PERK-mediated phosphorylation of the
Bcl-2 protein leading to the disruption of mitochondrial membrane
2
4
mustards in clinic. With this information in hand, it is necessary
to carry out chemical modifications on the nitrogen mustards to
enhance selectivity between normal and tumorous cells for the
chemotherapy of cancer. Recent studies have underpinned that
conjugating alkylating agents with an ideal natural compound of
⇑
Corresponding authors.
E-mail addresses: lzl1030@hotmail.com (Z. Li), huimhua@163.com (H. Hua),
lidahong0203@163.com (D. Li).
https://doi.org/10.1016/j.bmcl.2017.10.014
0
960-894X/Ó 2017 Published by Elsevier Ltd.
Please cite this article in press as: Hu X., et al. Bioorg. Med. Chem. Lett. (2017), https://doi.org/10.1016/j.bmcl.2017.10.014

2
X. Hu et al. / Bioorganic & Medicinal Chemistry Letters xxx (2017) xxx–xxx
special chemical scaffold is an attractive approach to enhance
activity and reduce adverse side effects of nitrogen mustard
2
5,26
drugs.
Previously, we synthesized a series of nitric oxide (NO) donating
2
2
derivatives of evodiamine and verified that the introduction of
the NO donating group to the N-13 position of evodiamine scaffold
significantly enhanced antitumor activity. So, in this letter, we also
designed and synthesized a series of evodiamine derivatives (bear-
ing nitrogen mustards) at N-13 according to combinational princi-
ple. Then, the antiproliferative activities of target compounds were
tested and preliminary mechanisms concerning the influence of
cell cycle progression, effects on cellular apoptosis and mitochon-
drial membrane potentials by representative compound 12c in
HL-60 cells were also disclosed.
The synthesis routine of nitrogen mustard-evodiamine hybrids
was illustrated in Scheme 1. First, 1 was treated with correspond-
ing bromohydrin (2-bromine ethanol, 3-bromine-1-propanol or 6-
bromine-1-hexanol) in the presence of NaH and anhydrous DMF as
1
9
reported to offer 2a–d in high yields. Esterification of 3 in dry
methanol in the presence of SOCl led to ester 4, and succedent
2
reaction of 4 with succinic anhydride produced melphalan deriva-
tive 5 in quantitative yield. The benzoic acid mustard 9 was pre-
2
7
pared according to the literature procedures. At last, the target
nitrogen mustard-evodiamine conjugates (10a–d, 11a–d and
1
2a–d) were synthesized from 2a–d accordingly with nitrogen
mustards (chlorambucil, 5 or 9) in the presence of DMAP/EDCI in
DCM at room temperature for 8–24 h. Each target compound28
was purified by column chromatography (PE/EA 7:1–2:1 v/v) and
1
13
confirmed by H NMR, C NMR and HR-MS (data in Supplemen-
tary Materials).
The antiproliferative activities of target hybrids 10–12 against
four different kinds of human cancer cell lines (PC-3 human pro-
static cancer cell line, HepG2 hepatocellular carcinoma cell line,
THP-1 human acute monocytic leukemia cell line, and HL-60
human leukemic cell line) and human normal peripheral blood
mononuclear cells (PBMC) were determined and compared with
lead compound evodiamine (1), evodiamine intermediates (2a–
d), nitrogen mustard compounds (chlorambucil, 5 and 9) and pos-
itive control 5-fluorouracil (5-FU) in each panel (Table 1). Com-
pounds 10c, 12b and 12c were very sensitive to HL-60 cell lines
with IC50 values ranging from 0.50 lM to 1.29 lM and even supe-
rior to 5-FU, and the other compounds were weaker than parent
compound 1. In THP-1 cell line, most hybrids showed the IC50 val-
ues at micromolar level, except for 11a. While, in PC-3 and HepG2
cell lines, target compounds showed almost no cytotoxicity and
only 12c exhibited moderate activity against HepG2 cells with
IC50 value of 17.04
pounds exhibited no antiproliferative activity (>200
l
M. In PBMC cells, all the target hybrid com-
M). The
l
antiproliferative selectivity between human tumorous and normal
blood cells was a very important property for the further exploita-
tion of antitumor drug. As shown in Table 1, generally, compounds
1
2a–d incorporated with melphalan showed stronger activity than
compounds 10a–d and 11a–d those with chlorambucil or benzoic
acid mustard against THP-1 and HL-60 cells. When R were (CH
and (CH -O-(CH groups (10b–c, 11b–c and 12b–c), the
antiproliferative activities were stronger than those of (CH and
groups (10a, 10d, 11a, 11d, 12a and 12d). The derivative
2c showed the most promising antiproliferative activity against
2 3
)
2
)
3
2 3
)
2 2
)
(
2 6
CH )
1
HL-60 cells with IC50 value of 0.5
investigation.
Cell cycle can be divided into four functional phases: S phase,
when DNA replication occurred; M phase (mitosis), when DNA
and cellular components are divided to form two daughter cells;
lM and selected for further
Scheme 1. Synthetic routine of target nitrogen mustard-evodiamine hybrids 10–
1
2. Reagents and conditions: (a) NaH, DMF, rt, 3 h; (b) SOCl
succinic anhydride, DMAP, DCM, rt, 18 h; (d) H SO , ethanol, reflux, 6 h; (e)
ethylene oxide, 25% aqueous acetic acid, rt, 24 h; (f) POCl , 50 °C, 0.5 h; 10% HCl, 12
h; (g) chlorambucil, 5 or 9, EDCI, DMAP, DCM, rt, 12–24 h.
2
, MeOH, reflux, 12 h; (c)
2
4
3
G
G
2
phase, between S and M phases, when cells prepare for mitosis;
phase, after mitosis and before S phase, when cells prepare for
1
2
9
another round of DNA and cellular replication. Many anticancer
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X. Hu et al. / Bioorganic & Medicinal Chemistry Letters xxx (2017) xxx–xxx
3
Fig. 1. The structures of reported evodiamine derivatives.
Table 1
Antiproliferative activity of target compounds 10–12 against four human cancer cell lines and human normal peripheral blood mononuclear cells (IC50
a
l
M).
Compd.
PC-3
HepG2
THP-1
HL-60
PBMC
1
13.97 ± 0.86
12.31 ± 0.70
27.26 ± 1.27
19.86 ± 1.38
21.48 ± 0.94
>100
63.46 ± 3.59
42.05 ± 2.33
45.69 ± 2.98
>100
27.62 ± 1.74
>100
>100
>100
>100
9.32 ± 0.59
>100
>100
>100
>100
13.83 ± 0.90
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
3.37 ± 0.11
1.64 ± 0.16
8.89 ± 0.60
8.42 ± 0.35
6.97 ± 0.41
3.84 ± 0.22
8.56 ± 0.36
5.30 ± 0.28
4.82 ± 0.45
5.68 ± 0.35
5.31 ± 0.25
10.52 ± 0.67
33.64 ± 1.92
6.60 ± 0.44
9.29 ± 0.48
3.92 ± 0.15
9.37 ± 0.64
2.37 ± 0.14
4.05 ± 0.22
4.24 ± 0.23
4.87 ± 0.29
1.39 ± 0.08
7.10 ± 0.15
6.03 ± 0.21
5.31 ± 0.28
8.53 ± 0.49
5.34 ± 0.28
12.29 ± 0.72
12.87 ± 0.65
11.28 ± 0.76
4.30 ± 0.24
1.29 ± 0.08
17.50 ± 1.64
27.25 ± 1.50
30.85 ± 1.96
22.33 ± 1.05
23.49 ± 0.75
3.11 ± 0.27
1.12 ± 0.07
0.50 ± 0.04
2.84 ± 0.71
3.69 ± 0.25
22.87 ± 1.52
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
>200
2
2
2
2
5
9
a
b
c
d
Chlorambucil
1
1
1
1
1
1
1
1
1
1
1
1
5
0a
0b
0c
0d
1a
1b
1c
1d
2a
2b
2c
2d
-FU
>100
>100
>100
>100
>100
23.94 ± 0.76
17.04 ± 0.84
>100
30.48 ± 1.20
b
NT
a
IC50: concentration that inhibits 50% of cell growth. Results are expressed as the mean ± S.D. of three independent experiments.
NT means not test.
b
compounds exerted their growth inhibitory effects either by
arresting the cell cycle at a particular checkpoint or by induction
of apoptosis or a combined effects of both cycle block and apopto-
sis. Furthermore, the regulation of the cell cycle and apoptosis was
considered to be effective cancer therapeutic methods.3 To reveal
the mechanism responsible for the antiproliferative effects of
nitrogen mustard-evodiamine hybrid derivatives, 12c was exam-
ined for the influence on the cell cycle progression. In this assay,
of apoptosis. Hoechst 33258, which stains the cell nuclei and emits
fluorescence allowing the visualization of nuclear morphological
changes, is a membrane permeable dye. Hence, we observed the
morphology of HL-60 cells stained with Hoechst, after treatment
0
with different concentrations (0, 0.25 lM, 0.5 lM and 1.0 lM) of
compound 12c for 48 h. As shown in Fig. 3, control cells were uni-
formly stained with Hoechst 33258 and presented round homoge-
neous nuclei, without morphological changes, while the 12c
treated HL-60 cells exhibited bright chromatin condensation and
nuclear fragmentation, a hallmark of apoptosis. These results indi-
cated that 12c could induce HL-60 cells apoptosis.
HL-60 cells were treated with 0.25 lM, 0.5 lM and 1.0 lM of com-
pound 12c. DMSO was used as vehicle. The DNA content of cell
nuclei was detected by flow cytometry. As shown in Fig. 2, the cells
in G
5.19% and 11.61%, respectively. After cells were treated with dif-
ferent concentrations of compound 12c for 48 h, the ratio of S
phase were almost not changed. The cells of G phase increased
to 13.19%, 15.19% and 25.99%, respectively. The results indicated
that 12c caused HL-60 cells arrest at G phase.
1
, S and G
2
phases of control group accounted for 63.21%,
Apoptosis is a process of programmed cell death and cancer
cells usually have an abnormal ability of proliferation mainly due
to the lack of apoptosis. Moreover, many compounds exert their
cytotoxic activity in tumor cells by inducing apoptosis. To clarify
whether the loss of cancer cell viability promoted by 12c was asso-
ciated with apoptosis, HL-60 cells were treated with vehicle alone
2
2
2
Hoechst 33258 staining was carried out to estimate whether
or different concentrations (0.25
lM, 0.5 lM and 1.0 lM) of 12c for
the growth inhibitory activities of 12c were related to the inducing
48 h and stained with FITC-Annexin V and propidium iodide (PI). PI
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X. Hu et al. / Bioorganic & Medicinal Chemistry Letters xxx (2017) xxx–xxx
Fig. 2. Effects on HL-60 cell cycle of 12c.
Fig. 4. Apoptosis effects of compound 12c in HL-60 cells.
Fig. 3. Hoechst staining of compound 12c in HL-60 cells.
Fig. 5. The effects of compound 12c on mitochondrial membrane potentials in HL-
is a DNA-intercalating dye which enters into the cells only when
plasma membrane integrity is lost, a typical event of late apoptotic
or necrotic cells. In the scatter plot of a double variant flow cytom-
etry, the lower left (LL) quadrant of the histograms represented the
viable cells, which excluded PI and were negative for FITC-Annexin
V staining. The lower right (LR) quadrant showed the early apop-
totic cells, which was PI negative and annexin V positive, indicating
that cell population endowed with cytoplasmic membrane integ-
rity. The upper right (UR) quadrant provided information about
the non-viable necrotic and late-stage apoptotic cells, which was
positive for annexin V binding and PI uptake. This population
had loss of membrane integrity and presented externalization of
PI. Finally, the upper left (UL) quadrant represented necrotic cell
population, which had loss of cell membrane integrity (PI positive
and Annexin-V negative).31 As shown in Fig. 4, 12c significantly
induced apoptosis in a dose-dependent manner. When treated
60 cells.
cle control, suggesting that 12c exerted its antiproliferative effects
possibly by inducing apoptosis.
The maintenance of mitochondrial membrane potential is sig-
nificant for mitochondrial integrity and bio-energetic function.
Mitochondrial changes, including loss of mitochondrial membrane
potential, are key events that took place during drug-induced apop-
tosis. On the other hand, it was reported that evodiamine might
induce apoptosis by activating the intrinsic mitochondria-related
pathway, which mainly resulted in the loss of mitochondrial mem-
5
,14
brane potential.
dria-mediated apoptosis in HL-60 cells by JC-1 method. HL-60
cells were incubated with different concentrations (0, 0.25 M,
0.5 M and 1.0 M) of compound 12c for 48 h prior to staining with
So, we studied the effects of 12c on mitochon-
l
l
l
with 0.25
percentages of apoptotic cells were 16.10%, 45.58% and 64.24%
early and late, Q2 + Q4), respectively, compared to 4.22% of vehi-
l
M, 0.5
lM and 1.0
lM of compound 12c for 48 h, the
the fluorescent dye JC-1. Then, the numbers of cells with collapsed
mitochondrial membrane potentials in different groups were
determined by flow cytometry analysis (Fig. 5). The percentage of
(
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X. Hu et al. / Bioorganic & Medicinal Chemistry Letters xxx (2017) xxx–xxx
5
apoptotic cells increased in a dose-dependent manner (7.81%,
4.13%, 28.51% and 62.78%, respectively). These results demon-
4. Shi L, Yang F, Luo F, et al. Tumour Biol. 2016;37:12791–12803.
5.
6.
7.
Wu WS, Chien CC, Chen YC, Chiu WT. PLoS One. 2016;11:e0160484.
Wang ZY, Liu JG, Li H, Yang HM. Am J Chin Med. 2016;44:1525–1541.
Fan X, Zhu JY, Sun Y, et al. Inflammation. 2017;40:1012–1027.
1
strated that the incubation of HL-60 cells with compound 12c
caused mitochondrial depolarization in the process of apoptosis.
In summary, twelve novel nitrogen mustard-evodiamine
hybrids (10–12) were designed and synthesized. The antiprolifera-
tive activities of all the target compounds were tested against
human cancer PC-3, HepG2, THP-1 and HL-60 cell lines and human
normal PBMC cells. Compound 12c showed potent cytotoxicity
against two tumor cell lines (THP-1 and HL-60), and even superior
to 5-fluorouracil. Preliminary SARs were also concluded based on
above results. 12c exhibited the most promising antiproliferative
8. Lv Q, Xue Y, Li G, et al. Int Immunopharmacol. 2015;28:1044–1049.
Kobayashi Y, Nakano Y, Kizaki M, Hoshikuma K, Yokoo Y, Kamiya T. Planta Med.
001;67:628–633.
9.
2
10. Ogasawara M, Matsunaga T, Takahashi S, Saiki I, Suzuki H. Biol Pharm Bull.
2002;25:1491–1493.
11. Shyu KG, Lin S, Lee CC, et al. Life Sci. 2006;78:2234–2243.
12. Yang J, Cai X, Lu W, et al. Cancer Lett. 2013;328:243–251.
13. Wei WT, Chen H, Wang ZH, et al. Int J Biol Sci. 2012;8:1–14.
14. Chen TC, Chien CC, Wu MS, Chen YC. Phytomedicine. 2016;23:68–78.
1
1
1
5. Sui H, Zhou LH, Zhang YL, et al. J Cell Biochem. 2016;117:1471–1481.
6. Jiang J, Hu C. Molecules. 2009;14:1852–1859.
7. Dong G, Sheng C, Wang S, Miao Z, Yao J, Zhang W.
2010;53:7521–7531.
J Med Chem.
activity with an IC50 value of 0.50 lM in HL-60 cells and good
1
1
2
8. Dong G, Wang S, Miao Z, et al. J Med Chem. 2012;55:7593–7613.
9. Song S, Chen Z, Li S, Huang Y, Wan Y, Song H. Molecules. 2013;18:15750–15768.
0. He S, Dong G, Wang Z, et al. ACS Med Chem Lett. 2015;6:239–243.
antiproliferative selectivity between tumorous and normal human
blood cells. Based on the above reasons, 12c was selected for fur-
ther mechanism study in HL-60 cell line. The results revealed that
incubation with representative compound 12c caused HL-60 cell
21. Wang S, Fang K, Dong G, et al. J Med Chem. 2015;58:6678–6696.
22. Zhao N, Tian KT, Cheng KG, et al. Bioorg Med Chem. 2016;24:2971–2978.
23. Povirk LF, Shuker DE. Mutat Res. 1994;18:205–226.
24. Chen W, Balakrishnan K, Kuang Y, et al. J Med Chem. 2014;57:4498–4510.
25. Liu K, Li DD, Zhao XM, Dai LL, Zhang T, Tao ZW. Appl Organomet Chem.
2016;1–7.
2
cycle arrest at G phase and induced apoptosis via mitochon-
drial-dependent pathways. Collectively, 12c might be considered
as a promising lead compound for the design of more efficacious
and less toxic novel anticancer agents.
2
2
2
6. Acharya PC, Bansal R. Steroids. 2017;123:73–83.
7. Zheng QZ, Zhang F, Cheng K, et al. Bioorg Med Chem. 2010;18:880–886.
8. Compound 12c: yellow oil, yield 64%. 1H NMR (CDCl
, 400 MHz), d (ppm) 8.11
(1H, d, J = 7.8 Hz, Ar-H), 7.60 (1H, d, J = 7.8 Hz, Ar-H), 7.47 (1H, m, Ar-H), 7.42
3
Acknowledgments
(
(
(
1H, m, Ar-H), 7.29 (1H, m, Ar-H), 7.22 (1H, m, Ar-H), 7.19 (1H, m, Ar-H), 7.15
1H, m, Ar-H), 6.97 (2H, d, J = 8.5 Hz, Ar-H), 6.59 (2H, d, J = 8.5 Hz, Ar-H), 6.17
This work was financially supported by Career Development
Support Plan for Young and Middle-aged Teachers in Shenyang
Pharmaceutical University.
1H, m, –NH), 5.99 (1H, s, NCH), 4.55 (1H, m, –CH), 4.28 (2H, m, –CH
), 3.73 (3H, s, –COOCH ), 3.69 (4H, m, NCH CH Cl), 3.60 (4H, m,
Cl), 2.90–3.37 (6H, m, –CH ), 2.49–2.63 (4H, m, –CH ), 2.39 (3H, s,
); C NMR (CDCl , 100 MHz) d (ppm) 172.78,
2
), 4.06 (2H,
m, –CH
NCH
NCH
2
3
2
2
2
CH
), 1.45–2.05 (8H, m, –CH
2
2
2
13
3
2
3
1
72.23, 171.00, 164.75, 151.07, 145.30, 137.37, 133.00, 131.04, 130.71 (Â2),
A. Supplementary data
129.01, 128.97, 125.86, 124.71, 124.21, 123.16, 122.72, 119.70, 119.09, 113.16,
1
4
12.17 (Â2), 109.95, 67.96, 67.64, 67.38, 65.70, 61.92, 53.59 (Â2), 53.43, 52.43,
0.70, 40.53 (Â2), 39.24, 36.83, 32.05, 30.90, 29.83, 29.05, 20.52; HR-MS (ESI, M
Supplementary data associated with this article can be found, in
the online version, at https://doi.org/10.1016/j.bmcl.2017.10.014.
+
2 5 7
H) m/z: calcd for C43H51Cl N O H: 820.3238, found: 820.3239. Spectrum data
of compounds 10a–c, 11a–c, 12a, 12b and 13a–c and experiment procedures of
MTT assay, cell cycle study, Annexin-V and PI double staining cellular apoptosis
analysis and mitochondrial membrane potential effects could be found in
Supplementary Materials.
Reference
2
9. Sánchez-Martínez C, Gelbert LM, Lallena MJ, de Dios A. Bioorg Med Chem Lett.
2015;25:3420–3435.
1.
2.
3.
Joshi P, Vishwakarma RA, Bharate SB. Eur J Med Chem. 2017;138:273–292.
Yu H, Jin H, Gong W, Wang Z, Liang H. Molecules. 2013;18:1826–1843.
Shi CS, Li JM, Chin CC, Kuo YH, Lee YR, Huang YC. Anticancer Res.
30. Blank M, Shiloh Y. Cell Cycle. 2007;6:686–695.
31. Raffa D, Maggio B, Plescia F, et al. Bioorg Med Chem. 2015;23:6305.
2
017;37:1149–1159.
Please cite this article in press as: Hu X., et al. Bioorg. Med. Chem. Lett. (2017), https://doi.org/10.1016/j.bmcl.2017.10.014