Anti-cancer effect of a novel 2,3-didithiocarbamate-substituted naphthoquinone as a tumor metabolic suppressor: In vitro and in vivo

Article abstract of DOI:10.1039/c8md00062j

Tumor cells reprogram their cellular metabolism by switching from oxidative phosphorylation to aerobic glycolysis to support aberrant cell proliferation. Suppressing tumor cell metabolism has become an attractive strategy for treating cancer patients. In this study, we identified a 2,3-didithiocarbamate-substituted naphthoquinone 3i that inhibited the proliferation of tumor cells by disturbing their metabolism. Compound 3i reduced cancer cell viability with IC₅₀ values from 50 nM to 150 nM against HCT116, MCF7, MDA-MB231, HeLa, H1299 and B16 cells. Further, compound 3i was found to suppress ATP production in cultured cancer cells, inhibit the M2 isoform of pyruvate kinase (PKM2) which is a rate-limiting enzyme in the glycolytic pathway and block the subsequent transcription of the downstream genes GLUT1, LDH and CCND1. In addition, exposure to compound 3i significantly suppressed tumor growth in a B16 melanoma transplantation mouse model and a spontaneous breast carcinoma mouse model in vivo. The identification of compound 3i as a tumor metabolic suppressor not only offers a candidate compound for cancer therapy, but also provides a tool for an in-depth study of tumor metabolism.

Full text of DOI:10.1039/c8md00062j

View Article Online
View Journal
MedChemComm
Accepted Manuscript
This article can be cited before page numbers have been issued, to do this please use: X. Ning, Y. Li, H.
Qi, R. Li, Y. Jin, J. Liu and Y. Yin, Med. Chem. Commun., 2018, DOI: 10.1039/C8MD00062J.
This is an Accepted Manuscript, which has been through the
Volume 7 Number 1 January 2016 Pages 1–204
Royal Society of Chemistry peer review process and has been
accepted for publication.
MedChemComm
Accepted Manuscripts are published online shortly after
acceptance, before technical editing, formatting and proof reading.
Broadening the field of opportunity for medicinal chemists
www.rsc.org/medchemcomm
Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available.
You can find more information about Accepted Manuscripts in the
author guidelines.
Please note that technical editing may introduce minor changes
to the text and/or graphics, which may alter content. The journal’s
Themed issue: Antibiotic Resistance
ISSN 2040-2503
standard Terms & Conditions and the ethical guidelines, outlined
in our author and reviewer resource centre, still apply. In no
event shall the Royal Society of Chemistry be held responsible
for any errors or omissions in this Accepted Manuscript or any
consequences arising from the use of any information it contains.
rsc.li/medchemcomm

Page 1 of 11
MedChemComm
View Article Online
DOI: 10.1039/C8MD00062J
Antiꢀcancer effect of a novel 2, 3ꢀdidithiocarbamate substituted
naphthoquinone as a tumor metabolic suppressor in vitro and
vivo
Xianling Ninga, Yunqiao Lia, c, Hailong Qia, b, Ridong Lia, Yan Jina, Junyi Liud* and
Yuxin Yina, b, c *
aInstitute of Systems Biomedicine, School of Basic Medical Sciences, Beijing Key Laboratory of
Tumor Systems Biology, Peking University Health Science Center, Beijing, China.
bPekingꢀTsinghua Center for Life Sciences, Peking University Health Science Center, Beijing,
China.
cDepartment of Pathology, School of Basic Medical Sciences, Peking University Health Science
Center, Beijing, China.
dState Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School
of Pharmaceutical Sciences, Peking University Health Science Center, Beijing, China.
ABSTRACT
Tumor cells reprogram their cellular metabolism by switching from
oxidative phosphorylation to aerobic glycolysis to support cell aberrant proliferation.
Suppressing tumor cell metabolism has become an attractive strategy for treating
cancer patients. In this study, we identified a 2, 3ꢀdidithiocarbamate substituted
naphthoquinone 3i that inhibited tumor cells proliferation by disturbing their
metabolism. Compound 3i reduced cancer cell viability with IC50 values from 50 nM
to 150 nM against HCT116, MCF7, MDAꢀMB231, Hela, H1299 and B16 cells.
Further, compound 3i was found to suppress ATP production in cultured cancer cells,
and inhibit M2 isoform of pyruvate kinase (PKM2) that is a rateꢀlimiting enzyme in
glycolytic pathway and block the subsequent transcription of the downstream gene
GLUT1, LDH and CCND1. In addition, exposure to compound 3i significantly
suppressed tumor growth in a B16 melanoma transplantation mouse model and a
spontaneous breast carcinoma mouse model in vivo. The identification of compound
3i as a tumor metabolic suppressor not only offers a candidate compound for cancer
therapy, but also provides a tool for an in depth study of tumor metabolism.
Key words
Metabolic suppressor, Compound 3i, Antiproliferative activity, Suppressing tumor
growth
1. Introduction
Cancer cells differ from most normal cells in metabolism. The normal cells rely
generally on mitochondrial oxidative phosphorylation to generate energy from
* Corresponding author
Email address: jyliu@bjmu.edu.cn
* Corresponding author
Email address: yinyuxin@hsc.pku.edu.cn
1

MedChemComm
Page 2 of 11
View Article Online
DOI: 10.1039/C8MD00062J
glucose, whereas cancer cells instead rely on glycolysis. 1ꢀ4. This difference suggests
that targeting tumor metabolism could be a selective approach to suppress tumor
growth.5ꢀ7 Glucose goes through a series of biochemical transformations with
production of ATP in the process of glycolysis, and each step is catalyzed by specific
enzymes.5 Influencing these enzymes to reduce or reverse the abnormal
reprogrammed metabolism of cancer cells is an attractive therapeutic strategy for
cancer patients. The pyruvate kinase (PK) is a rateꢀlimiting enzyme that regulates the
final step in glycolysis and catalyzes the transfer of a phosphate group from
phosphoenolpyruvate (PEP) to adenosine diphosphate (ADP) to yield pyruvate and
adenosine triphosphate (ATP)8, 9. There are four isoforms of PK (M1, M2, L and R) in
mammalian cells: the M1 isoform (PKM1) is expressed in many differentiated tissues,
PKM2 is expressed during embryonic development and over expressed in tumor
10ꢀ12
tissues, PKL and PKR are expressed in liver and erythrocytes, respectively
.
Many studies have shown that tumorigenesis is connected with the reꢀexpression of
PKM2 together with a downꢀregulation of the expression of PKM1 and other
isozymes.9, 13 In addition, PKM2 activates βꢀcatenin to induce CCDN1 and cꢀMyc
expression and upregulate GLUT1 and lactate dehydrogenase A (LDHA)14ꢀ17
.
Upregulation of these glycolysis genes increases glucose intake, consumption and
lactate production to promote tumorigenesis 1.
We previously synthesized a series of dithiocarbamate substituted naphthoquinone
derivatives and evaluated their antiꢀproliferative effects in the cellular level and
PKM2 inhibition activity in enzyme level.18, 19 To further improve the activity, we
continue to optimize the structure. In this study, we synthesized a new 2,
3ꢀdidithiocarbamate substituted naphthoquinone compound 3i (Fig. 1A), which
exhibited more potent antiꢀproliferative effect than previously synthesized compounds.
It reduced cancer cell viability with IC50 values from 50 nM to 100 nM against
HCT116, MCF7, MDAꢀMB231, Hela, H1299 and B16 cells. We also found that
compound 3i suppressed tumor cells metabolism by reducing ATP production in
cancer cells, and inhibiting PKM2 activity and blocking the subsequent transcription
of the downstream gene GLUT1, LDH and CCND1. In addition, exposure to
compound 3i significantly suppressed tumor growth in a B16 melanoma
transplantation mouse model and a spontaneous breast carcinoma mouse model in
vivo.
2. Materials and methods
2.1 Synthesis of compound 3i
2.1.1 Procedure for preparation of 2,3ꢀbisꢀchloromethylꢀ[1,4]naphthoquinone
The 1,4ꢀnaphthaquinone (1 g, 6.3 mmol) in glacial acetic acid (20 mL) was taken in a
100 mL roundꢀbottomed flask, and 36% aqueous formaldehyde (6 mL) was added.
The reaction solution was cooled in iceꢀwater. Dry hydrogen chloride passed in for 2
h. The solution became red, then being kept at room temperature for 48 h. The
reaction mixture was poured on ice and extracted with ethyl acetate. The combined
organic fractions were washed with brine, dried (Na2SO4), and concentrated under
reduced pressure. Purification of the crude residue by column chromatography
2

Page 3 of 11
MedChemComm
View Article Online
DOI: 10.1039/C8MD00062J
(petroleum ether/ethyl acetate) afforded the title compound (yellow solid). The yield
1
of this reaction was 68.9%. H NMR (400 MHz, CDCl3) δ 8.18ꢀ8.20 (m, 2H, ArH),
7.81ꢀ7.83 (m, 2H, ArH), 4.72 (s, 4H, 2CH2Cl).
2.1.2 Procedure for preparation of dipiperidineꢀdithiocarbamic acid
3ꢀdipiperidinethiocarbamoylsulfanylmethylꢀ1,4ꢀdioxoꢀ1,4ꢀdihydroꢀnaphthalenꢀ2ꢀylme
thyl ester (3i)
Carbon disulfide (180 ꢁL, 3 mmol) and piperidine (297 ꢁL, 3 mmol) were added to
CH3CN (5 mL) and the resulting solution was stired for 30 minutes. 2,
3ꢀBisꢀchloromethylꢀ[1,4]naphthoquinone (254 mg, 1 mmol) was added in portions at
frequent intervals. Then the reaction mixture was kept at room temperature for 48 h.
The reaction mixture was concentrated in vacuo, diluted with H2O, and extracted with
CH2Cl2. The combined organic fractions were washed with brine, dried (Na2SO4), and
concentrated under reduced pressure. Purification of the crude residue by column
chromatography (petroleum ether/ CH2Cl2) afforded compound 3i (yellow solid). The
1
yield of this reaction was 83.5%. Mp 142ꢀ143 °C. H NMR (400 MHz, CDCl3) δ
8.11ꢀ8.13 (m, 2H, ArH), 7.73ꢀ7.75 (m, 2H, ArH), 4.86 (s, 4H, 2CH2S), 4.29 (q, 4H,
2NCH2), 3.87 (q, 4H, 2NCH2), 1.70 (m, 12H, 2CH2CH2CH2). 13C NMR (100 MHz,
CDCl3) δ 194.21, 183.95, 143.97, 133.84, 132.05, 126.65, 34.11, 24.25. HRꢀMS (ESI+)
m/z: 505.1112 [M+H]+, 527.0931[M+Na]+. Found: 505.1100 [M+H]+, 527.0903
[M+Na]+.
2.2 Cell culture
Cell lines were grown with routine culture techinques in RPMI 1640 supplemented
with 9% fetal bovine serum at 37°C in 5% CO2.
2.3 MTS cell proliferation assay
Cells were plated in 96ꢀwell plates at a density of 5000ꢀ10000 cells per well. 12 h
after seeding, cells were treated with various concentrations of test compounds for 48
h. Cell viability was assessed with the MTS assay (Promega) according to the
manufacturer's instruction.
2.4 Measurement of ATP
HCT116 cells were seeded into 6ꢀwell plates at a density of 5×105 cells per well. 24 h
after seeding, cells were treated with 10 ꢁM or 20 ꢁM compound 3i for 6 h. ATP
levels were measured using the Cell TiterꢀGlo Luminescent Cell Viability Assay
(Promega).
2.5 PKM2 activity assay
Pyruvate kinase activity was measured with a fluorescent pyruvate kinaseꢀlactate
dehydrogenase coupled assay as previously described 20. To evaluate PKM2 activity
in cell lysates, HCT116 cells were treated with various concentrations of compound 3i
for 8 h and lysed in NP40 lysis buffer immediately before measuring pyruvate kinase
activity as described previously 21.
2.6 Quantitative realꢀtime PCR (RTꢀqPCR)
HCT116 cells were seeded into 6ꢀwell plates at a density of 5×105 cells per well. 24 h
after seeding, cells were treated with 20 ꢁM compound 3i for 8 h. Total RNA was
extracted using TRIzol (Invitrogen). cDNA synthesis was carried out using a cDNA
synthesis kit (Tansgene). qPCR was then carried out using SYBR Green Master Mix
3

MedChemComm
Page 4 of 11
View Article Online
DOI: 10.1039/C8MD00062J
(Transgene) in a bioꢀred Realꢀtime PCR machine. PCR primers sequences used are
listed in Supplementary Table S1.
2.7 B16 melanoma transplantation mouse model
Female C57BL/6 mice were injected with 1×106 B16 cells subcutaneously in the
armpits. Approximately 6 days later, B16 tumor appeared, and mice were paired (N =
7) and injected with compound 3i (25 mg/kg and 50 mg/kg) or vehicle. Compound 3i
was dissolved in 5% (v/v) DMAC (dimethylacetamide) and added to olive oil.
Intraperitoneal injection was performed in the mouse. The animals were injected once
every two days. Tumor volume was calculated using the following equation:
V=L(S2)π/6, where L is the longer and S is the shorter of the two tumor dimensions.
2.8 Spontaneous breast carcinoma mouse model
C57BL/6 spontaneous breast carcinoma mice 22 were paired (N = 3) and injected with
compound 3i (50 mg/kg) or vehicle, when spontaneous tumor volumes reached 0.1
cm3. Intraperitoneal injection was performed in the mouse. Animals were injected
once every two days.
All experiments were performed in compliance with the relevant laws and
institutional guidelines of the Institute Research Ethics Committee of Peking
University Health Science Center, and the committee had approved the experiments.
2.9 Statistical analysis
Statistical analysis was performed using GraphPad Prism 5.0. Data are presented as
mean ± SD (n = 3).
3. Results and discussion
We prepared compound 3i by the synthetic route as shown in Figure 1B. To determine
the efficiency of compound 3i as an antiꢀtumor agent, we assessed the in vitro
cytotoxicity of 3i using several different tumor cell lines derived from human colon
cancer (HCT116), breast cancer (MCF7), breast cancer (MDAꢀMB231), cervical
cancer (Hela) and lung cancer (H1299), and mouse melanoma (B16). The results are
presented in Table 1. Compound 3i reduced cancer cell viability at nanomolar
concentrations with IC50 values against HCT116, MCF7, MDAꢀMB231, Hela, H1299
and B16 cells from 50 nM to 150 nM in MTS reduction assays. Specially, compound
3i exhibited a doseꢀdependent cytotoxicity (Fig. 2). To further explore the selectivity
of the target compound against cancer cells, we tested its cytotoxicity in BEASꢀ2B
cells derived from normal human bronchial epithelial cells. As seen in Table 1,
compound 3i showed higher selectivity for cancer cells H1299 than normal cells
BEASꢀ2B, which indicated that compound 3i probably have low toxicity to normal
cells.
Compound 3i
4

Page 5 of 11
MedChemComm
View Article Online
DOI: 10.1039/C8MD00062J
Fig. 1. The structure and synthetic route of compound 3i. (A) The structure of compound 3i. (B)
The synthetic route of compound 3i. Reagents and conditions: (a) formaldehyde, HCl, HAc, H2O,
0
°C, 68.9%; (b) CS2, piperidine, CH3CN, rt, 83.5%.
Table 1. In vitro cytotoxicity of compound 3i
IC50 ± SD (ꢁM)
0.077 ± 0.011
0.061 ± 0.030
0.135 ± 0.010
HCT116
MCF7
MDAꢀMB231
Hela
0.124 ± 0.010
H1299
0.109 ± 0.015
0.104 ± 0.007
32.61 ± 2.04
B16
BEASꢀ2B
Fig. 2. Compound 3i reduces cancer cell viability. Cells were treated with increasing
concentrations of compound 3i. Cell viability was measured using MTS.
The direct consequence of inhibiting tumor cells metabolism is the decrease of ATP
production in cells. We treated HCT116 cells with 10 ꢁM and 20 ꢁM compound 3i
and tested ATP production with the ATP Assay Kit. The results showed that compound
3i significantly influenced the metabolic function by impairing the cellular ATP
production (Fig. 3A). Compound 3i has a similar napthaquinone skeleton as in
shikonin23 that was reported to affect the metabolism of tumor cells by inhibiting
PKM2 activity, therefore, we tested the influence of compound 3i on PKM2 activity
using a fluorescent PKꢀLDH coupled assay according to a previously reported method
5

MedChemComm
Page 6 of 11
View Article Online
DOI: 10.1039/C8MD00062J
20. Shikonin was used as the positive control. As shown in Table 2, compound 3i (IC50
= 0.88±0.37) displayed the higher inhibitory activity than shikonin (IC50 = 8.82 ±
2.62). Moreover, compound 3i showed inhibition of PKM2 with less inhibition of
PKM1 and PKL. To investigate whether compound 3i is able to inhibit PKM2 in cells,
we treated cells in which PKM2 is highly expressed with 2.5 ꢁM, 5 ꢁM, 10 ꢁM and
20 ꢁM compound 3i and assayed PKM2 activity in the corresponding cell lysates. We
found that compound 3i inhibited PKM2 activity in cells in a dose dependent manner
(Fig. 3B). In addition, there are extensive evidences that PKM2 coactivates βꢀcatenin
to induce its downstream gene CCND1 and cꢀMyc transcription, resulting in
upregulation of GLUT1 and LDHA. We evaluated the transcription of these genes.
Cells were treated with 20 ꢁM compound 3i, and RTꢀPCR showed significant
reduction of GLUT1, LDH and CCND1 (Fig. 3C). These data suggest that
compound 3i probably interferes with energy metabolism of cancer cells by inhibiting
PKM2 activity.
A
B
C
Fig. 3. Compound 3i can regulate the metabolism of tumor cells. (A) Compound 3i inhibited
cellular ATP production. HCT116 cells were treated with 10 ꢁM and 20 ꢁM compound 3i for 6 h
and ATP production was tested with the ATP Assay Kit. (B) Compound 3i inhibited PKM2 activity
in cells in a dose depend manner. HCT116 cells were treated with 2.5 ꢁM, 5 ꢁM, 10 ꢁM and 20
ꢁM compound 3i for 8 h and PKM2 activity was tested in the corresponding cell lysates using a
6

Page 7 of 11
MedChemComm
View Article Online
DOI: 10.1039/C8MD00062J
fluorescent PKꢀLDH coupled assay. (C) Compound 3i inhibited the transcription of
PKM2ꢀregulated glycolytic genes GLUT1, LDH and CCND1. HCT116 cells were treated with 20
ꢁM compound 3i for 8 h and qPCR was performed.
Table 2. In vitro inhibitory activity (IC50) of 3i and shikonin on different PKM2 isoforms
PKM2 (ꢁM)
PKM1 (ꢁM)
PKL (ꢁM)
IC50(PKM1)/
IC50(PKM2)
5.8
IC50(PKL)/
IC50(PKM2)
3.3
3i
0.88 ± 0.37
8.82 ± 2.62
5.07 ± 0.13
2.90 ± 0.64
Shikonin
12.96 ± 3.37
39.25 ± 6.53
1.5
4.5
Based on the potent inhibitory effect of compound 3i on tumor cells in vitro, we next
assessed its inhibitory efficiency in mouse models. The in vivo antiꢀtumor effect of
compound 3i was first evaluated in a B16 transplantation mouse model. As shown in
Fig. 4A, intraperitoneal injection of compound 3i at 25 and 50 mg/kg every two days
for a period of 12 days demonstrated significant doseꢀdependent inhibition of tumor
growth over the course of the treatment. The mice treated with compound 3i at 50
mg/kg showed tumors of 30% tumor weight compared to the vehicleꢀtreated (control)
group (Fig. 4B). The T/C values (relative tumor volume growth rate) of the 50 mg/kg
treatment group were close to or less than 40% at each time point, which consistent
with the high efficiency of compound 3i. (Fig. 4D). In addition, compound 3i had no
significant effect on body weight over the course of this experiment, suggesting that it
was well tolerated in vivo.
Fig. 4. Compound 3i inhibited B16 tumor growth in a dose depend manner. (A) Mice were treated
with 25 mg/kg or 50 mg/kg compound 3i or vehicle. Tumor volume was measured once every two
7

MedChemComm
Page 8 of 11
View Article Online
DOI: 10.1039/C8MD00062J
days. (B) The weight of individual tumors was measured. (C-D) The T/C values of the 25 mg/kg
and 50 mg/kg treatment group at selected time points.
To further assess the therapeutic efficiency of this compound, we used a PTEN
deleted mediated mouse spontaneous breast tumor model22. Mice were divided into
two groups when tumors reached a volume of 0.1 cm3, and compound 3i at 50 mg/kg
or vehicle was injected intraperitoneally every two days for 36 days. As shown in Fig.
5A, tumor growth was significantly suppressed over this course of treatment. Mouse
tumor weight after treatment with compound 3i was approximately 20% of that of the
control (Fig. 5B and 5C).
Fig. 5. Compound 3i significantly inhibited mouse spontaneous breast tumor growth. (A) Mice
were treated with compound 3i or vehicle. Tumor volume was measured once every four days.
(B-C) Tumors were removed after sacrificing mice. Weight of individual tumors was measured.
4. Conclusions
Modulation of metabolism is a key characteristic of highly proliferative cancer cells
which allows both rapid ATP generation and access to metabolites needed as cellular
building blocks. Here, we describe a metabolic suppressor compound 3i that is a
previously unreported compound. Compound 3i reduced cancer cell viability in a high
response with IC50 values in nanomolar concentrations and suppressed ATP
production in cancer cells. Meanwhile, compound 3i inhibited a rateꢀlimiting enzyme
PKM2 in glycolytic pathway. However, the cytotoxicity of compound 3i was much
8

Page 9 of 11
MedChemComm
View Article Online
DOI: 10.1039/C8MD00062J
higher than PKM2 inhibitory activity, which suggested that compound 3i had other
mechanisms to influence tumor cells metabolism and suppress cell proliferation. The
1,4ꢀnaphthoquinone moiety of compound 3i has been reported to influence other
proteins, such as, DTꢀdiaphorase24 and P2X7 receptor25, which may lead to the poor
activity relationship in the enzyme and cell level. However, there is no doubt that
compound 3i targets PKM2 and affects tumor cells metabolism. In future study, we
will focus on investigating other mechanisms of compound 3i.
In this study, we also found compound 3i significantly inhibited PTEN loss mediated
mouse spontaneous breast tumor growth. Previous studies have indicated that
PTENꢀnegative human hepatocellular carcinoma cell lines show upꢀregulation of
PKM2 expression, which is advantageous for cell proliferation and
anchorageꢀindependent growth26. This probably accounts for the fact that the
compound 3i is very sensitive to PTEN loss in the spontaneous tumor model. The
mechanism will be investigated in our further work. In addition, we will also continue
optimization of the structure of 2, 3ꢀdidithiocarbamate substituted naphthoquinones
and prepare suitable formulation for identifying new compounds with better
physicoꢀchemical properties and higher efficiency.
Conflict of Interest
The authors declare no competing interest.
Acknowledgments
This study was supported by the National Natural Science Foundation of China (Key
grants #81430056, #81372491 and #81402777) and the China Postdoctoral Science
Foundation (#2014M560026 and #2015T80028).
Reference
1.
W. Yang and Z. Lu, Cancer Lett, 2013, 339, 153ꢀ158.
A. T. Ooi and B. N. Gomperts, Clinical cancer research : an official journal of the
American Association for Cancer Research, 2015, 21, 2440ꢀ2444.
J. Penkert, T. Ripperger, M. Schieck, B. Schlegelberger, D. Steinemann and T. Illig,
Oncotarget, 2016, 7, 67626ꢀ67649.
2.
3.
4.
5.
6.
O. Warburg, Science, 1956, 123, 309ꢀ314.
Y. Zhao, E. B. Butler and M. Tan, Cell death & disease, 2013, 4, e532.
N. D. Amoedo, E. Obre and R. Rossignol, Biochimica et biophysica acta, 2017, 1858,
674ꢀ685.
7.
8.
N. E. Scharping and G. M. Delgoffe, Vaccines, 2016, 4.
N. Wong, J. De Melo and D. Tang, International journal of cell biology, 2013, 2013,
242513.
9.
V. Gupta and R. N. Bamezai, Protein Sci, 2010, 19, 2031ꢀ2044.
W. Luo and G. L. Semenza, Trends in endocrinology and metabolism: TEM, 2012, 23,
560ꢀ566.
10.
11.
T. Noguchi, H. Inoue and T. Tanaka, The Journal of biological chemistry, 1986, 261,
13807ꢀ13812.
9

MedChemComm
Page 10 of 11
View Article Online
DOI: 10.1039/C8MD00062J
12.
13.
W. Yang and Z. Lu, J Cell Sci, 2015, 128, 1655ꢀ1660.
H. R. Christofk, M. G. Vander Heiden, M. H. Harris, A. Ramanathan, R. E. Gerszten, R.
Wei, M. D. Fleming, S. L. Schreiber and L. C. Cantley, Nature, 2008, 452, 230ꢀ233.
W. Yang, Y. Xia, H. Ji, Y. Zheng, J. Liang, W. Huang, X. Gao, K. Aldape and Z. Lu,
Nature, 2011, 480, 118ꢀ122.
14.
15.
16.
17.
T. L. Dayton, T. Jacks and M. G. Vander Heiden, EMBO Rep, 2016, 17, 1721ꢀ1730.
X. Gao, H. Wang, J. J. Yang, X. Liu and Z. R. Liu, Mol Cell, 2012, 45, 598ꢀ609.
X. Gao, H. Wang, J. J. Yang, J. Chen, J. Jie, L. Li, Y. Zhang and Z. R. Liu, The Journal of
biological chemistry, 2013, 288, 15971ꢀ15979.
18.
19.
20.
21.
22.
X. Ning, H. Qi, R. Li, Y. Li, Y. Jin, M. A. McNutt, J. Liu and Y. Yin, Eur J Med Chem,
2017, 138, 343ꢀ352.
X. Ning, H. Qi, R. Li, Y. Jin, M. A. McNutt and Y. Yin, J Enzyme Inhib Med Chem, 2018,
33, 126ꢀ129.
M. G. Vander Heiden, H. R. Christofk, E. Schuman, A. O. Subtelny, H. Sharfi, E. E.
Harlow, J. Xian and L. C. Cantley, Biochemical pharmacology, 2010, 79, 1118ꢀ1124.
H. R. Christofk, M. G. Vander Heiden, N. Wu, J. M. Asara and L. C. Cantley, Nature,
2008, 452, 181ꢀ186.
A. Di Cristofano, B. Pesce, C. CordonꢀCardo and P. P. Pandolfi, Nature genetics, 1998, 19,
348ꢀ355.
23.
24.
25.
J. Chen, J. Xie, Z. Jiang, B. Wang, Y. Wang and X. Hu, Oncogene, 2011, 30, 4297ꢀ4306.
C. Flader, J. Liu and R. F. Borch, J Med Chem, 2000, 43, 3157ꢀ3167.
R. X. Faria, F. H. Oliveira, J. P. Salles, A. S. Oliveira, N. L. von Ranke, M. L. Bello, C. R.
Rodrigues, H. C. Castro, A. R. Louvis, D. L. Martins and V. F. Ferreira, Eur J Med Chem,
2018, 143, 1361ꢀ1372.
26.
I. Nemazanyy, C. Espeillac, M. Pende and G. Panasyuk, Biochemical Society transactions,
2013, 41, 917ꢀ922.
10

Page 11 of 11
MedChemComm
View Article Online
DOI: 10.1039/C8MD00062J
Graphical abstract:
O
O
S
N
S
S
N
S
Compound 3i