Brief Article

pubs.acs.org/jmc

A Rhodium(III)-Based Inhibitor of Lysine-Specific Histone

Demethylase 1 as an Epigenetic Modulator in Prostate Cancer Cells
Chao Yang,†,∥ Wanhe Wang,‡,∥ Jia-Xin Liang,†,∥ Guodong Li,† Kasipandi Vellaisamy,‡
Chun-Yuen Wong,§ Dik-Lung Ma,*,‡ and Chung-Hang Leung*,†
†
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau,
China
‡
Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China
§
Department of Biology and Chemistry, City University of Hong Kong, Tat Chee Avenue, Kowloon, Hong Kong SAR, China
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*
S Supporting Information

ABSTRACT: We report herein a novel rhodium(III) complex 1 as a new

LSD1 targeting agent and epigenetic modulator. Complex 1 disrupted the
interaction of LSD1-H3K4me2 in human prostate carcinoma cells and
enhanced the amplification of p21, FOXA2, and BMP2 gene promoters.
Complex 1 was selective for LSD1 over other histone demethylases, such as
KDM2b, KDM7, and MAO activities, and also showed antiproliferative activity
toward human cancer cells. To date, complex 1 is the first metal-based inhibitor
of LSD1 activity.

■ INTRODUCTION
The post-translational modification of histones to regulate gene
attractive route to new therapies for various cancers, and
research into new LSD1 inhibitors is an active area of research.
expression via the modeling of nucleosome structure is a central Metal complexes have attracted considerable attention as
theme in epigenetics. 1,2 This “histone code” includes anticancer agents in recent decades.23,24 Although metal
methylation, acetylation, phosphorylation, and ubiquitina- complexes tend not to be readily orally available, they possess
other benefits compared to purely organic molecules, making
tion.3,4 The presence of histone lysine demethylases suggests
them appealing alternatives for therapeutic agent develop-
that histone lysine methylation is a reversible epigenetic
modification.4,5 ment.25,26 Metal complexes can exhibit various shapes depend-
Lysine-specific demethylase 1 (LSD1) catalyzes the deme- ing on the coordination number of the metal complex and the
type of ligands.27 Recent interest has focused on the discovery
thylation of di- and monomethylated Lys4 of histone H3 but
of molecularly targeted metal against specific enzymes or
not trimethylated H3K4.6,7 LSD1 activity is central to the
protein−protein interactions.28−34 In particular, rhodium(III)
development and maintenance of acute myeloid leukemia
complexes have been discovered to inhibit STAT3, β-amyloid
(AML),8,9 as well as prostate carcinoma, colon carcinoma
fibrillation, and JAK2.35−37 Moreover, metal-based epigenetic
cancer, and breast cancer.10,11 Aberrant activity of LSD1 is
modulators have been very recently discovered.25,38 However,
linked with tumor suppressor gene silencing, promoting
there are no metal-based inhibitors of LSD1 reported to date.
tumorigenesis.12,13 Thus, considerable effort has been devoted
We report herein a novel rhodium(III) complex 1 as the first
to developing LSD1 inhibitors for the treatment of cancer,
metal-based inhibitor of LSD1 activity and that shows promise
despite a lack of precise mechanistic insight.14 Several types of
for the treatment of prostate cancer.

LSD1 inhibitors have been presented in the literature, including
peptides, natural products and their derivatives, and poly-
amines.15−19 Wang and co-workers have reported CBB1007
(6),20 a known potent, reversible, and substrate-competitive
LSD1 inhibitor in F9 cells. ORY-1001 (9),21 a potent and
selective inhibitor of LSD1, has entered phase I clinical trials for
the treatment of AML, while 4-[[4-[[[(1R,2S)-2-phenyl-
cyclopropyl]amino]methyl]piperidin-1-yl]methyl]benzoic acid
(GSK2879552),22 an orally available, irreversible inhibitor of
LSD1, has also recently entered phase I trials for AML and
small-cell lung cancer therapy. Inhibition of LSD1 offers an
■
RESULTS AND DISCUSSION
Synthesis and Characterization of Metal-Based Com-
plexes. To develop potential metal-based scaffolds as LSD1
inhibitors, rhodium(III) complexes 1−5 were synthesized
(Figure 1) bearing structurally diverse C∧N and N∧N ligands.
Complex 1 bears the 4-chloro-2-phenylquinoline C∧N ligand,
Received: January 25, 2017
Published: February 20, 2017

© 2017 American Chemical Society 2597 DOI: 10.1021/acs.jmedchem.7b00133

J. Med. Chem. 2017, 60, 2597−2603
Journal of Medicinal Chemistry
Figure 1. Chemical structures of rhodium(III) complexes 1−5, 6, 7
(4-chloro-2-phenylquinoline), 8 (4,4′-dimethoxy-2,2′-bipyridine), and
9 evaluated in this study.
while complex 2 carries the related 2-phenylquinoline ligand.
Complexes 3 and 5 bear the relatively small 2-phenylpyridine
C∧N ligand, while complex 4 has the 1-phenyl-1H-pyrazole
C∧N ligand. With regard to the N∧N ligand, complexes 1 and 5
carry a 2,2′-bipyridine scaffold substituted with methoxy
groups, respectively, while complex 2 carries the unsubstituted
2,2′-bipyridine ligand. Finally, complex 4 bears the 2,9-
dimethyl-1,10-phenanthroline N∧N ligand, while complex 3
contains the same scaffold substituted with pendent phenyl
groups. Complex 3 has been previously investigated for its
anticancer and antibacterial activity, whereas complexes 4 and 5
have been studied for their ability to inhibit BRD4, an
epigenetic “reader” protein.31,39 Complexes 1 and 2 are novel
and are reported for the first time in this work.
Inhibition of LSD1 Activity in Vitro. We screened
racemic complexes 1−5 for their ability to modulate LSD1
activity using a fluorescence-based assay. Complexes 1−5 have
no intrinsic fluorescence (Figure S1 in Supporting Informa-
tion). From those results, the rhodium(III) complex 1 showed
the greatest inhibition of LSD1, as it reduced LSD1 activity by a
fluorescence-based method at 10 μM (Figure 2a). Compound 6
Figure 2. (a) Complexes 1−6 inhibit the activity of LSD1 as
determined using a fluorimetric assay. (b) Complex 1 dose inhibits
LSD1 activity as determined by a fluorimetric assay. Error bars
represent the standard deviations of results obtained from three
independent experiments.
reduced LSD1 activity by nearly 74.7% at the same
concentration.20 Complexes 2, 4, and 5 showed moderate
inhibitory activity in this assay, while complex 3 was nearly
inactive. A dose−response experiment was then performed to
determine the efficacy of complex 1 against LSD1 demethylase
activity. Complex 1 inhibited LSD1 activity in a dose-
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dependent manner, with an IC50 of 0.04 ± 0.008 μM (Figure
2b). Furthermore, we performed a kinetic assay to determine
the kinetics of inhibition. A kinetic assay showed that like the
competitive inhibitor 6,31 complex 1 acts as a competitive
inhibitor of LSD1 with Ki of 0.57 μM (Figure S2).
Structure−Activity Relationships. To further investigate
the mechanism of action of 1, we evaluated the activity of its
isolated ligands 7 and 8 (Figure 1). The results showed that
neither 7 nor 8 had any effect on the activity of LSD1 (Figure
S3). This indicates that the isolated ligands are not effective
against LSD1 activity on their own. Additionally, a preliminary
structure−activity relationship analysis can be performed by
considering the biological potency of complexes 1−5. Complex
2 was the second-most active member of this series, which
could be possibly accounted for by the observation that the 2-
phenylquinoline C∧N ligand of 2 is simply the dechlorinated
derivative the 4-chloro-2-phenylquinoline C∧N ligand of 1. This
indicates that C∧N ligands of that approximate size might have
higher complementarity with the binding site of LSD1
compared to C∧N ligands of other sizes. Finally, the weakest
activity of complex 5 indicates that the combination of the 2-
phenylpyridine C∧N ligand and the 4,4′-dimethoxy-2,2′-
bipyridine N∧N ligand is undesirable for biological activity.
Molecular Docking Analysis. To further investigate the
mechanism of action of complex 1, we evaluated the binding
mode of complex 1 to LSD1 via molecular docking. Docking
was performed using the Molsoft ICM method (ICM-Pro 3.6-
1d molecular docking software). A peptide substrate-like
H3K4M, which resembles the natural histone H3 peptide
except where lysine 4 (K4) is replaced by methionine, is
predicted to bind across the entire LSD1 (PDB code 2V1D)
binding site, forming H-bonding interactions with Asp375 and
Asp 556 (Figure S4A). This peptide inspired the design of 6,
which is also predicted to bind across most of the site (Figure
S4B). In contrast, 9 is predicted to be located only on the lower
half of the binding site (Figure S4C). In this respect, the
predicted binding mode of complex 1 resembles 9 more than 6
or the substrate-like peptide, as complex 1 occupies only part of
the LSD1 binding site, contacting the residues around Asp556
(Figures 3 and S4D).
Selective Inhibition of the LSD1 in Human Prostate
Carcinoma Cells. Previous studies have revealed that LSD1 is
most likely responsible for the gene inactivation of p21.40,41
Moreover, inactivation of LSD1 significantly induced the
expression of differentiation genes such as FOXA2,
Figure 3. Top view of complex 1 bound to the LSD1 generated by
molecular docking. LSD1 (PDB code 2V1D) is depicted in ribbon
form and is colored purple. Complex 1 is depicted as a space-filling
representation showing carbon (yellow), oxygen (red), nitrogen
(blue), and chloride (green) atoms. The binding pocket of the
LSD1 is represented as a translucent green surface.
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BMP2.42,43 Chromatin immunoprecipitation (ChIP)-qPCR
assays were performed to test whether complex 1 could block
the binding of LSD1 to chromatin in human prostate cancer
PC3 cells and human prostate carcinoma epithelial 22RV1 cells,
thereby relieving the suppression of downstream genes. Cells
were treated with the indicated concentration of complex 1,
and cell lysates were collected and immunoprecipitated with
anti-H3K4me2 antibody. ChIP-qPCR analysis showed that
complex 1 could increase the amplification of the p21, FOXA2,
and BMP2 gene promoters (Figures 4 and S5). This result
Figure 4. Effect of complex 1 on the level of H3K4me2 at the p21,
FOXA2, BMP2, and GAPDH genes in PC3 cells. ChIP-qPCR assays
were performed with primary antibody against H3K4me2. Error bars
represent the standard deviations of results obtained from three
independent experiments.
suggested that complex 1 could suppress the demethylation of
H3K4me2 on these promoters in PC3 cells or 22RV1 cells,
thus leading to increased gene expression.
Complex 1 induced the accumulation of H3K4me2 in treated
PC3 cells in a dose-dependent manner (Figure S6), which we
presume is due to its effect on LSD1 activity. To further
evaluate the mechanism of action of complex 1, coimmuno-
precipitation experiments were performed. Cell lysates were
immunoprecipitated with LSD1 antibody and immunoblotted
with the appropriate antibodies. Notably, complex 1
interrupted the LSD1-H3K4me2 interaction in PC3 (Figure
5) and 22RV1 (Figure S7) cells, which could account for its
Figure 5. Effect of 1 on the interaction between LSD1 and other
proteins in PC3 cells studied by coimmunoprecipitation. PC3 cells
were treated with the indicated concentrations of 1 or 6 for 24 h.
Protein lysates were incubated with anti-LSD1 magnetic beads, and
precipitated proteins were analyzed by Western blotting with the
indicated antibodies.
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ability to induce H3K4me2 accumulation as described
previously. However, complex 1 had no effect on the binding
between LSD1 and H3K4, H3K4me3, REST, and CoREST in
PC3 cells (Figure 5). As REST and CoREST form part of the
LSD1-containing epigenetic complex,44,45 this suggests that
complex 1 does not act by disrupting the interactions between
LSD1 and REST or CoREST.
In view of the promising activity of complex 1 at binding to
and inhibiting the activity of LSD as described above, the
specific mechanism of complex 1 was further investigated using
an enzyme-linked immunosorbent assay. In this assay, PC3 cells
were treated with different concentration of complex 1 for 24 h,
and cell lysates were harvested. In this assay, active LSD1 in the
cell lysates binds to and demethylates a unique dimethylated
histone H3K4 substrate. Then, remaining nondemethylated
substrate is recognized with a high affinity antimethylated
histone H3K4 antibody. The results showed that complex 1
decreased H3K4me2 demethylation with an IC50 of 0.23 μM
(Figure 6A), while the IC50 of 6 in the same assay was 7.23 μM.
Figure 6. Selectivity of complex 1 for histone demethylases in vitro.
PC3 cell lysates were collected after treatment of cells with complex 1
for 24 h. Complexes 1 and 6 dose-dependently inhibit LSD1 (A)
activity in PC3 cells as determined by ELISA. Effect of complex 1 on
KDM2b (B), KDM7 (C), MAO (D) activity in PC3 cells as
determined by ELISA. Error bars represent the standard deviations of
results obtained from three independent experiments.
These results suggest that complex 1 could inhibit the activity
of LSD1 in vitro with stronger potency compared to 6 and are
consistent with the results of the previous experiments.
To further investigate the specificity of complex 1 for LSD1,
we examined the activity of complex 1 against the
demethylation of other histone demethylases targeting di- or
monomethylated histone H3. KDM2b, highly expressed in
human leukemia cells, not only catalyzes the demethylation of
mono- and dimethylated H3K26 but also demethylates tri- and
dimethylated H3K4.46 Another JmjC domain containing
protein, KDM7, catalyzes demethylation of both mono- and
dimethylated H3K9 and H3K27.47 Thus, PC3 cells were
incubated with various concentrations of complex 1 for 24 h,
and the demethylase activities of the cell lysates were
investigated using ELISA. Complex 1 showed only weak
potency against the activities of the other demethylases, with
only 10% inhibition at the highest concentration (10 μM)
tested (Figure 6B,C). This indicates that complex 1 is selective
for LSD1 over other demethylase enzymes, which could be
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expected since these demethylases have a different mechanism
and are not flavin-dependent.
LSD1 belongs to the flavin adenine dinucleotide (FAD)-
dependent amine oxidases superfamily, while the monoamine
oxidases (MAOs) belong to the same superfamily and share the
same enzymatic mechanism of LSD1 demethylase.48 Therefore,
the enzyme activity observed activity of complex 1 against
MAO was measured on the same cell treatment as before by
monitoring the fluorescence at excitation of 530 nm and
emission of 585 nm. In brief, MAO reacts with p-tyramine, a
substrate for MAOA and MAOB, resulting in the formation of
H2O2, which is determined by a fluorimetric method. Complex
1 exhibited little or no inhibition of MAOA/B in PC3 cells
(Figure 6D). Taken together, these results together indicate
that complex 1 is selective for LSD1 over other related
enzymes, including KDM2b, KDM7, and MAO.
Inhibition of GLUT1 Expression in Human Prostate
Carcinoma Cells. Glucose transporter 1 (GLUT1) is a
uniporter carrier protein that exhibits variable expression in
various tumor types.49 Recent studies have suggested that
LSD1 is linked with the increased expression of GLUT1 in
human cancer cells.50 Therefore, the effect of complex 1 on the
expression of GLUT1 was further explored. PC3 cells were
treated with the indicated concentration of complex 1 for 24 h,
and GLUT1 levels were analyzed by Western blotting. The
results showed that complex 1 could decrease GLUT1
expression and have no effect on the expression of LSD1 and
GAPDH in PC3 cells (Figure 7).
Figure 7. Effect of complex 1 on GLUT1 protein expression in PC3
cells. PC3 cells were treated with the indicated concentrations of
complex 1 for 24 h, and then GLUT1 levels were analyzed by Western
blotting.
A previous study has reported that GLUT1 expression was
reduced in knockdown MAOA cells under hypoxia.51 There-
fore, we performed a quantitative PCR experiment to study the
impact of complex 1 on LSD1 or MAOA knockdown cells
(Figure S8A). Knockdown of LSD1 and MAOA resulted in a
decrease of GLUT1 mRNA levels by 40% and 20%, respectively
(Figure S8B). However, cells were more resistant to 1 in LSD1
knockdown cells when compared with knockdown cells with
control or MAOA siRNA (Figure S8C). Taken together, these
data suggest that 1 suppresses GLUT1 at the transcriptional
level in a manner that is, at least in part, LSD1 dependent.
Complex 1 Inhibits Cellular Proliferation and Induces
Cell Cycle Arrest. The antiproliferative activities of 1 and 6
against 22RV1 cells, human prostate cancer DU145 cells,
human breast adenocarcinoma MCF-7 cells, PC3 cells, human
epithelial colorectal adenocarcinoma Caco2 cells, human
embryonic kidney 293 cells, and human liver LO2 cells were
assessed by the XTT assay. Cell viability was measured after 72
h of treatment with complex 1. IC50 values of complex 1 were
greater than 100 μM against PC3 cells, 22RV1 cells, or LO2
cells (Figure S9). By comparison, complex 1 was most potent
against human prostate cancer PC3 cells, with an IC50 of 2.83
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μM (Figure S10). Moreover, the toxicity of 1 was significantly
reduced in LSD1-knockdown PC3 cells (Figure S10). This
indicates that complex 1 could be considered as a potential
scaffold for the development of antiprostate cancer drugs.
To evaluate the nature of cell death induced by complex 1,
we evaluated the levels of proteins related to apoptosis,
including cleaved caspase 3 and cleaved PARP, in the presence
of 1. Complex 1 had no effect on the expression level of
apoptosis markers in PC3 cancer cells and LO2 normal cells
(Figure S11A). In flow cytometry analysis, a minor population
of complex-1-treated cells were observed in the Q1-LR quarter
(early apoptosis: FITC annexin V positive and PI negative)
(Figure S11B), indicating that complex 1 below 3 μM, as well
as 6 at 1 μM, had no effects on early apoptosis in PC3 and LO2
cells. Moreover, the expression of cyclin D1, a protein related to
G0/G1 cell cycle arrest, was down-regulated after complex 1
treatment (Figure S11A). Furthermore, complex 1 increased
cells in the G0/G1 phase and reduced cell counts in the S phase
in a dose-dependent manner, indicating that complex 1 could
induce G0/G1 arrest in PC3 and LO2 cell lines (Figure S11C
and Figure S11D). Taken together, apoptosis and cell cycle
arrest analysis suggested that complex 1 could induce G0/G1
arrest, but not apoptosis, in PC3 cancer and LO2 normal cell
lines.
■ CONCLUSIONS
In summary, we have described a novel rhodium(III) complex
1 as the first metal-based inhibitor of LSD1 activity. Complex 1
exhibited superior inhibitory activity against LSD1 in a
fluorescence-based assay and an ELISA assay compared to
the known LSD1 inhibitor, 6. Moreover, complex 1 reduced the
proliferation of human prostate cancer PC3 cells at low
micromolar concentrations. Additionally, complex 1 enhanced
the amplification of LSD1-regulated promoters and decreased
the LSD1-H3K4me2 interaction in PC3 cells. Complex 1 also
selectively inhibited LSD1 activity without inhibiting the
activity of other related enzymes, KDM2b, KDM7, and
MAO. Finally, complex 1 inhibited the expression of GLUT1
in PC3 cells, presumably due to its ability to inhibit LSD1
activity. We anticipate that complex 1 could be considered as a
useful scaffold for the further development of more selective
and potent epigenetic modulators to treat different types of
cancer, including prostate cancer.
■
EXPERIMENTAL SECTION
General Synthesis of [Rh2(C∧N)4Cl2] complexes. Cyclometa-
lated dichloro-bridged dimers of the general formula [Rh2(C∧N)4Cl2]
were synthesized according to a literature method.52 In brief, RhCl3·
xH2O was heated to 140 °C with 2.1 equiv of cyclometallated C∧N
ligands in 3:1 methoxymethanol and deionized water under a nitrogen
atmosphere overnight. The reaction was cooled to room temperature,
and the product was filtered and washed with three portions of
deionized water and then three portions of ether to yield the
corresponding dimer.
General Synthesis of [Rh(C∧N)2(N∧N)]PF6 Complexes. These
complexes were synthesized using a modified literature method.52
Briefly, a suspension of [Rh2(C∧N)4Cl2] (0.1 mmol) and correspond-
ing N∧N (0.21 mmol) ligands in a mixture of dichloromethane/
methanol (1:1, 6 mL) was refluxed overnight under a nitrogen
atmosphere. The resulting solution was allowed to cool to room
temperature and was filtered to remove unreacted cyclometallated
dimer. To the filtrate, an aqueous solution of ammonium
hexafluorophosphate (excess) was added, and the filtrate was reduced
in volume by rotary evaporation until precipitation of the crude
DOI: 10.1021/acs.jmedchem.7b00133
J. Med. Chem. 2017, 60, 2597−2603
Journal of Medicinal Chemistry
product occurred. The precipitate was then filtered and washed with
several portions of water (2 × 30 mL) followed by diethyl ether (2 ×
30 mL). The solid was dissolved into acetone, and then the product
was precipitated by adding diethyl ether and filtered. The purity of all
complexes was determined by an Agilent 1200 high-performance
liquid chromatography (HPLC) system. The results showed that the
purity of all complexes was over 95% (Figure S12).
Complex 1. Yield: 49%. HPLC purity: >95%. 1H NMR (400 MHz,
acetone-d6) δ 8.72 (s, 2H), 8.38 (d, J = 7.9 Hz, 2H), 8.26−8.15 (m,
4H), 7.89 (d, J = 2.6 Hz, 2H), 7.72 (d, J = 8.8 Hz, 2H), 7.60 (t, J = 8.2
Hz, 2H), 7.31 (t, J = 8.7 Hz, 2H), 7.26 (t, J = 8.0 Hz, 2H), 7.20 (dd, J
= 6.3, 2.6 Hz, 2H), 6.94 (t, J = 7.8 Hz, 2H), 6.64 (d, J = 7.6 Hz, 2H),
3.94 (s, 6H); 13C NMR (101 MHz, acetone) δ 169.56, 169.07, 167.16,
156.79, 149.97, 148.32, 146.52, 145.72, 136.13, 132.50, 131.45, 128.88,
128.41, 126.61, 125.96, 124.85, 119.45, 119.43, 114.62, 110.96, 57.03.
MALDI-TOF-HRMS calcd for C42H30Cl2N4O2Rh [M − PF6]+:
795.0801. Found: 794.9014. Anal. (C42H30Cl2F6N4O2PRh + H2O)
C, H, N: calcd 52.57, 3.36, 5.84; found 52.66, 3.23, 5.98.
Complex 2. Yield: 35%. HPLC purity: >95%. 1H NMR (400 MHz,
acetone-d6) δ 8.64−8.51 (m, 4H), 8.45 (d, J = 5.2 Hz, 2H), 8.40−8.27
(m, 4H), 8.10 (t, J = 7.9 Hz, 2H), 7.94 (d, J = 8.1 Hz, 2H), 7.64 (t, J =
7.6 Hz, 2H), 7.57 (d, J = 8.9 Hz, 2H), 7.44 (t, J = 8.0 Hz, 2H), 7.25 (t,
J = 8.0 Hz, 2H), 7.15 (t, J = 8.7 Hz, 2H), 6.92 (t, J = 7.5 Hz, 2H), 6.61
(d, J = 7.8 Hz, 2H); 13C NMR (101 MHz, acetone) δ 167.35, 155.37,
149.10, 147.67, 146.59, 140.82, 140.68, 136.03, 131.46, 131.09, 130.00,
129.10, 128.51, 128.06, 127.76, 125.86, 124.82, 124.52, 119.16.
MALDI-TOF-HRMS calcd for C40H28N4Rh [M − PF6]+: 667.1369.
Found: 667.1373. Anal. (C40H28F6N4PRh + 2H2O) C, H, N: calcd
56.62, 3.80, 6.60; found 56.40, 3.52, 6.61.
Complex 3. HPLC purity: >95% (reported).39
Complex 4. HPLC purity: >95% (reported).31
Complex 5. HPLC purity: >95% (reported).31
■
ASSOCIATED CONTENT
*
S Supporting Information
The Supporting Information is available free of charge on the
ACS Publications website at DOI: 10.1021/acs.jmed-
chem.7b00133.
Experimental methods and supplemental data (PDF)
Molecular formula strings (CSV)
■ AUTHOR INFORMATION
Corresponding Authors
*D.-L.M.: e-mail, edmondma@hkbu.edu.hk; phone, +852
9251-0870.
*C.-H.L.: e-mail, duncanleung@umac.mo; phone, +853 8822-
4688.
ORCID
Chun-Yuen Wong: 0000-0003-4780-480X
Dik-Lung Ma: 0000-0002-9515-340X
Chung-Hang Leung: 0000-0003-2988-3786
Author Contributions
∥ (8) Harris, W. J.; Huang, X.; Lynch, J. T.; Hitchin, J. R.; Li, Y. Y.;
C.Y., W.W., and J.-X.L. contributed equally to this work. C.Y.,
W.W., J.L., and G.L. carried out all the experiments. C.Y., W.W.,
and K.V. wrote the manuscript. D.-L.M., C.-Y.W., and C.-H.L
analyzed the results. C.-H.L. and D.-L.M. designed the
experiments.
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
This work is supported by Hong Kong Baptist University
(Grant FRG2/15-16/002), the Health and Medical Research
Fund (Grant HMRF/14130522), the Research Grants Council
2601
Brief Article
(Grants HKBU/12301115, HKBU/204612, and HKBU/
201913), the French National Research Agency/Research
Grants Council Joint Research Scheme (Grant A-HKBU201/
12, Oligoswitch), National Natural Science Foundation of
China (Grant 21575121), Guangdong Province Natural
Science Foundation (Grant 2015A030313816), Hong Kong
Baptist University Century Club Sponsorship Scheme 2016,
Interdisciplinary Research Matching Scheme (Grant RC-
IRMS/15-16/03), the Science and Technology Development
Fund, Macao SAR (Grant 098/2014/A2), the University of
Macau (Grants MYRG2015-00137-ICMS-QRCM,
MYRG2016-00151-ICMS-QRCM, and MRG044/LCH/2015/
ICMS), and National Natural Science Foundation of China
(Grant 21628502).
■
ABBREVIATIONS USED
LSD1, lysine-specific histone demethylase 1; AML, acute
myeloid leukemia; H3K4me2, histone H3 dimethyl Lys4;
MAO, monoamine oxidase; MAOA, monoamine oxidase A;
MAOB, monoamine oxidase B; ELISA, enzyme-linked
immunosorbent assay; DMEM, Dulbecco’s modified Eagle
medium; FBS, fetal bovine serum; PBS, phosphate buffered
saline; PC3 cell, human prostate carcinoma cell; Caco2 cell,
human epithelial colorectal adenocarcinoma cell; LO2 cell,
human liver cell; 293 cell, human embryonic kidney cell;
22RV1 cell, human prostate carcinoma epithelial cell; DU145
cell, human prostate cancer cell; KDM2b, lysine-specific histone
demethylase 2b; KDM7, lysine-specific histone demethylase 7;
XTT, cell proliferation kit II; H3K26me2, histone H3 dimethyl
Lys26; H3K27me2, histone H3 dimethyl K27
■
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2603 DOI: 10.1021/acs.jmedchem.7b00133

J. Med. Chem. 2017, 60, 2597−2603