Design, synthesis and biological evaluation of quinoline derivatives as HDAC class I inhibitors

Article abstract of DOI:10.1016/j.ejmech.2017.03.064

Inhibition of histone deacetylase (HDAC) has been regarded as a potential therapeutic approach for treatment of multiple diseases including cancer. Based on pharmacophore model of HDAC inhibitors, a series of quinoline-based N-hydroxycinnamamides and N-hydroxybenzamides were designed and synthesized as potent HDAC inhibitors. All target compounds were evaluated for their in?vitro HDAC inhibitory activities and anti-proliferative activities and the best compound 4a surpass Vorinostat in both enzymatic inhibitory activity and cellular anti-proliferative activity. In terms of HDAC isoforms selectivity, compounds 4a exhibited preferable inhibition for class I HDACs, especially for HDAC8, the IC₅₀ value (442?nM) was much lower than that of Vorinostat (7468?nM). Subsequently, we performed class I & IIa HDACs whole cell enzyme assay to evaluate inhibitory activity in whole cell context. Compounds 4a and 4e displayed much better cellular activity for class I HDACs than that for class IIa HDACs, which indicated that 4a and 4e might be potent class I HDAC inhibitors. Meanwhile, flow cytometry analysis showed that compound 4a and 4e can promote cell apoptosis in?vitro.

Full text of DOI:10.1016/j.ejmech.2017.03.064

Accepted Manuscript
Design, synthesis and biological evaluation of quinoline derivatives as HDAC class I
inhibitors
Chen Chen, Xuben Hou, Guohua Wang, Wenyan Pan, Xinying Yang, Yingkai Zhang,
Hao Fang
PII:
S0223-5234(17)30231-3
10.1016/j.ejmech.2017.03.064
EJMECH 9324
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 1 February 2017
Revised Date: 23 March 2017
Accepted Date: 25 March 2017
Please cite this article as: C. Chen, X. Hou, G. Wang, W. Pan, X. Yang, Y. Zhang, H. Fang, Design,
synthesis and biological evaluation of quinoline derivatives as HDAC class I inhibitors, European Journal
of Medicinal Chemistry (2017), doi: 10.1016/j.ejmech.2017.03.064.
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ACCEPTED MANUSCRIPT

ACCEPTED MANUSCRIPT
Design, synthesis and biological evaluation of quinoline derivatives as HDAC class I
inhibitors
Chen Chen a, Xuben Hou a,b, Guohua Wang a, Wenyan Pan a, Xinying Yang a, Yingkai Zhangb, Hao
Fang a*
a
Department of Medicinal Chemistry, Key Laboratory of Chemical Biology (Ministry of Education),
School of Pharmaceutical Sciences, Shandong University, 44, West Culture Road, 250012, Jinan,
Shandong, PR China
b Department of Chemistry, New York University, New York, New York 10003 United States
*Address correspondence to this author:
Prof. Hao Fang, Department of Medicinal Chemistry, School of Pharmacy, Shandong University,
44 West Culture Road, Jinan, PR China.
E-mail: haofangcn@sdu.edu.cn
Tel. 86-531-88382019
Fax. 86-531-88382548

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Abstract:
Inhibition of histone deacetylase(HDAC) has been regarded as a potential therapeutic approach
for treatment of multiple diseases including cancer. Based on pharmacophore model of HDAC
inhibitors, a series of quinoline-based N-hydroxycinnamamides and N-hydroxybenzamides were
designed and synthesized as potent HDAC inhibitors. All target compounds were evaluated for
their in vitro HDAC inhibitory activities and anti-proliferative activities and the best compound 4a
surpass Vorinostat in both enzymatic inhibitory activity and cellular anti-proliferative activity. In
terms of HDAC isoforms selectivity, compounds 4a exhibited preferable inhibition for class I
HDACs, especially for HDAC8, the IC50 value (442 nM) was much lower than that of Vorinostat
(7468 nM). Subsequently, we performed class I & IIa HDACs whole cell enzyme assay to
evaluate inhibitory activity in whole cell context. Compounds 4a and 4e displayed much better
cellular activity for class I HDACs than that for class IIa HDACs, which indicated that 4a and 4e
might be potent class I HDAC inhibitors. Meanwhile, flow cytometry analysis showed that
compound 4a and 4e can promote cell apoptosis in vitro.
Keyword:
Quinoline
Hydroxamic acid
Class I HDACs
Anti-proliferative
Class I celluar activity
Pro-apoptosis activity
1. Introduction
At present, cancer has been a leading cause of death and a major public health problem with
increasing incidence and mortality. It has been widely believed that widespread epigenetic
dysregulation that interacts extensively with underlying genetic mutations lead to tumorigenesis[1].
Histone deacetylases (HDACs) and histone acetyltransferases (HATs) played a pivotal role in
epigenetic regulation of gene expression and had pleiotropic effects at the cellular and systemic
levels[2]. HDACs removed the acetyl groups of lysine residues, leading to chromatin structure
becoming contraction and therewith repressing transcription. HATs opposed the effect of HDACs,
which referred to histone acetylation and relaxed chromatin structure resulting in enhancing
transcription [3]. There was a balance for histone acetylation and deacetylation between the
activities of both HATs and HDACs in normal cells. Human HDACs had 18 members and they
were divided into four classes. Class I HDACs acted a vital part in tumorigenesis through forming
complexes with other proteins. For example, HDAC1 connected with retinoblastoma tumor
suppressor protein and formed complex, which was a key controlling factor on cell proliferation
and differentiation. The transcriptional repressor complex, producing by association of HDAC2
with YY1, a mammalian zinc-finger transcription factor, played an essential role in transcriptional

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regulation[4]. HDAC8 has been proved to be important for cancer cell survival, when HDAC8
was knocked down, the growth of certain cancer cell lines (HeLa and HCT116) was suppressed[5].
HDAC8 expression level was also positively correlated to poor prognosis and metastasis of
neuroblastoma[6]. Thus, HDAC inhibitors (HDACIs) have capabilities to control acetylation level
of histones and non-histone proteins by inhibiting HDAC enzymes, which then produce a variety
of effects on cancer cells, such as cancer cell arrest, differentiation and active cell apoptosis [7].
Attributed to decades of synthetic efforts, five HDAC inhibitors (Vorinostat, Romidepsin,
Belinostat, Panobinostat and Chidamide) have been applied to the clinical therapy (Fig. 1) [8] and
more than twenty candidates have been initiated in clinical trials in monotherapy or combination
therapy. Vorinostat, Romidepsin, Belinostat and Panobinostat were approved by US FDA for
treatment of cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL) and
multiple myeloma (MM), respectively[9-12], while Chidamide has been only approved in
China[8]. Based on literature investigations, most HDAC inhibitors possessed a three-motif
pharmacophore model, comprising of a zinc binding group (ZBG), a linker group and a surface
recognition group (cap group) [13]. The ZBGs chelated zinc ion within HDACs active site and
were indispensable for HDAC inhibitory activities. The linker group bound into the acetyl-lysine
binding channel and connected the ZBG with the cap group, which further enhanced the
HDAC-inhibitor interactions.
Fig. 1 Approved HDAC inhibitors.
Lots of HDAC inhibitors have been developed by utilizing different heterocycles as the cap
group, such as thiadiazole[14, 15], saccharin[16], purine[17, 18]，N-phenylquinazolin[19] and
quinoline[20]. Importantly, quinoline hydroxamic acid derivatives has been proved to exert
HDAC inhibitory activities and anti-proliferative activities in our previous study [20]. Meanwhile,
N-hydroxycinnamamide and N-hydroxybenzamide, which serve as both ZBGs and linker groups,
were widely used fragments in current HDACIs design[21, 22]. Three HDACIs (pracinostat,
belinostat and panobinostat) with N-hydroxycinnamamide fragment have been approved by FDA
and several N-hydroxybenzamide HDACIs are in clinical trials[23]. Guided by these structural
analyses above, we developed a series of new HDACIs by utilizing quinoline derivatives as cap
group and utilized N-hydroxycinnamamide and N-hydroxybenzamide as the ZBG and linker
groups. This article reports the synthesis and preliminary biological evaluation of these
compounds.

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2. Results and discussion
2.1. Chemistry
The routes used for synthesis of the entire target compounds are outlined in Schemes 1-3. As
shown in Scheme 1, starting materials 1a-1g and 2a-b were commercially available or easily
synthesized according to literature [20, 24]. Compounds 1a-1g reacted with 2a-2b according to
reductive amination to obtain 3a-3g. Compounds 3a-3g were directly converted to
N-hydroxycinnamamides 4a-4g by treating with hydroxylamine. In order to increase the structural
diversity, N-alkylation were performed to introduce R1 group to get 5a-5g, which were converted
to N-hydroxycinnamamides 6a-6g. As shown in Scheme 2, 9a-9b were easily synthesized by
Wittig-Horner reaction. Reductive amination of 1a-1g with substituted aniline yielded 7a-7g,
which next did coupling reaction with 9a or 9b to get 10a-10i. Compounds 10a-10i were next
converted to N-hydroxycinnamamides 11a-11i. As shown in Scheme 3, final
N-hydroxybenzamides 14a-14d and 16a were synthesized following the similar procedures as
shown in Scheme 1.
Scheme 1. Synthesis of target compounds 4a-g and 6a-g. Reagents and conditions: (a) MeOH,
65°C, 1h; NaBH4, rt, 1h; (b) NH2OH•HCl, KOH, MeOH, rt, 4 h; (c) R1-X，DMSO, DIEA, 50°C,
2-4h; (d) NH2OH•HCl, KOH, MeOH, rt, 4 h.
Scheme 2. Synthesis of target compounds 11a-11i. Reagents and conditions: (a) (i)aniline, formic
acid, DCM, rt, overnight; (ii) NaBH4, MeOH, rt, 1h; (b) trimethyl phosphonoacetate, K2CO3, H2O,

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rt, overnight; (c) (i) oxalyl chloride, DCM, rt, 1h (ii)NaHCO3, DCM, rt, 4h (d) NH2OH•HCl, KOH,
MeOH, rt, 4 h.
Scheme 3. Synthesis of target compounds 14a-d and 16a. Reagents and conditions: (a) MeOH,
65°C, 1h; NaBH4, rt, 1h; (b) NH2OH•HCl, KOH, MeOH, rt, 4 h; (c) DMSO, DIEA, 50°C, 2-4h; (d)
NH2OH•HCl, KOH, MeOH, rt, 4 h.
2.2. Hela cell extract inhibitory assay
HDAC inhibitory activities of all the target compounds and the positive control drug Vorinostat
were evaluated by the Color de Lys™ assay (BMLAK501, Enzo®Life Sciences) including
HDAC1&2. Results were summarized as IC50 values in Table 1.
As shown in Table 1, the HDAC inhibitory activities of N-hydroxycinnamamides 4a-4f were
superior to N-hydroxybenzamides 14a-14d. As far as N-hydroxycinnamamides were concerned,
the HDAC inhibitory activities were influenced by the position of substituents on benzene ring A.
For example, compounds with para-substituted N-hydroxycinnamamide exhibited better activities
than meta-substituted N-hydroxycinnamamide derivatives. The differences in linker groups also
had significant impacts on inhibitory activities against HDACs. Compounds 11a-11i with amide
groups between quinoline and N-hydroxycinnamamide showed lower activities than compounds
4a-4f that with amine groups.
Besides, substituents on quinoline ring and amine also had effects on the inhibitory activities
against HDACs. Substitution on amine (R1 groups) decreased the HDAC inhibitory activities. For
example, the IC50 values of compounds 4a-4g were lower than that of compounds 6a-6g (>1000
nM). On the other hand, substituents on quinoline ring (R2 and R3 groups) only showed slight
effects on potency and the IC50 values of compounds 4d, 4e, 4f all distributed between 200-300
nM. Different from our previous study, in which the substituents on the R2, R3-position of
quinoline enhanced the enzymatic inhibitory activities[20], compound 4a without any substituent
in R1，R2，R3 manifested best enzymatic inhibitory activity with a IC50 value of 138 nM against
HDAC1&2.

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Table 1
The chemical structures and HDACs inhibitory activities of quinoline hydroxamate derivatives.
Cpd.
R1
R2
R3
Linker
Substitution
IC50 (nM)
of
A
H
H
Cl
H
H
CH2
CH2
para
para
para
para
para
para
para
para
para
para
para
meta
meta
meta
para
para
para
para
para
para
para
para
para
138±7
305±47
375±55
236±76
275±64
206±51
＞1000
＞1000
＞1000
＞1000
＞1000
559±84
＞1000
＞1000
＞1000
＞1000
744±48
＞1000
＞1000
724±14
720±97
＞1000
560±187
4a
4b
4c
H
H
Br
MeO
H
H
CH2
H
H
CH2
4d
4e
H
H
Cl
MeO
H
CH2
H
CH2
4f
Bn
H
CH2
6a
Bn
Cl
H
CH2
6b
6c
Bn
Br
MeO
Br
Br
Br
Br
H
H
CH2
Bn
H
CH2
6d
6e
n-hexyl
H
H
CH2
H
CH2
4g
Bn
H
CH2
6f
n-hexyl
Ph
H
CH2
6g
H
C=O
11a
11b
11c
11d
11e
11f
11g
11h
11i
4-Br-Ph
4-MeO-Ph
4-MeO-Ph
Ph
H
H
C=O
H
H
C=O
H
Cl
H
C=O
Br
H
C=O
Ph
MeO
H
C=O
Ph
H
(C=O) CH2
(C=O) CH2
(C=O) CH2
Ph
Br
MeO
H
Ph
H
Cpd.
R1
R2
R3
IC50 (nM)
H
H
H
H
H
Cl
H
H
H
＞1000
＞1000
＞1000
＞1000
836±63
14a
14b
14c
14d
16a
H
Br
H
MeO
H
Bn
159±36
Vorinostat

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2.3 In vitro anti-proliferative assay
Compounds 4a, 4b, 4d, 4e and 4f that possessed higher HDAC inhibitory activities than others
were selected to evaluate their anti-proliferative activities in vitro. MTT assays were performed
using five different cancer cell lines including MOLT-4 (acute lymphoblastic leukemia cell), HEL
(erythrocyte leukemic cell), PC-3 (prostatic cancer cell), K562 (chronic myelogenous leukemia
cell) and Hela (cervical carcinoma cell). The IC50 values were summarized in Table 2. Results
indicated that all tested compounds showed better anti-proliferative activities in different cancer
cell lines compared with Vorinostat. Specially, compound 4e exhibited the highest
anti-proliferative activities in all five cancer cell lines.
Table 2 Anti-proliferative activity of compounds 4a, 4b, 4d, 4e and 4f
Cpd.
IC50a (µM)
MOLT-4
0.24±0.01
0.25±0.02
0.23±0.01
0.16±0.02
0.19±0.02
0.56±0.04
HEL
K562
PC-3
Hela
0.15±0.01
0.22±0.02
0.13±0.01
0.11±0.01
0.22±0.04
0.25±0.02
0.51±0.01
0.72±0.04
0.69±0.01
0.36±0.02
0.75±0.03
1.95±0.26
3.08±0.69
4.35±0.35
3.70±0.60
2.92±0.35
4.60±0.50
13.91±0.73
2.20±0.43
3.03±0.13
2.68±0.08
1.82±0.16
3.33±0.61
3.55±0.15
4a
4b
4d
4e
4f
Vorinostat
a IC50 were expressed as the mean ± standard deviation of two or three separate determinations
2.4 HDAC isoforms inhibitory activity of representative compounds
To explore the HDAC isoforms inhibitory profile, we chose compounds 4a with most potent
HDACs inhibitory activity and 4e with highest anti-proliferative activity to perform enzyme
inhibitory assays against class I HDACs (HDAC1 and HDAC8) and class II HDAC (HDAC6).
Results in Table 3 indicated that compound 4a and 4e possessed better inhibitory activities against
class I HDACs (HDAC1 and HDAC8) than Vorinostat. For HDAC1, compounds 4a and 4e
exhibited approximate inhibitory activity with their IC50 values slight lower than Vorinostat. For
HDAC8, compounds 4a and 4e exposed notably inhibitory activity, particularly IC50 value of 4a
reached to the 500nM range while IC50 value of Vorinostat remained approximately 8µM.
Table 3 HDAC isoforms inhibitory activity of compounds 4a and 4e
a
Cpd.
IC50 (nM)
HDAC6
HDAC1
72.5±0.5
95.0±1.0
HDAC8
442.0±72.0
911.0±32.0
7468.5±559.5
49.5±2.5
36.5±5.5
14.5±2.5
4a
4e
Vorinostat 106.0±0.0
a IC50 were expressed as the mean ± standard deviation of two or three separate determinations
2.5 HDAC class I and class IIa whole cell assay
Subsequently, compounds 4a and 4e were chosen to perform HDAC whole cell enzyme assay
to examine their inhibitory activities as well as and isoforms selectivity in whole cell context.
Firstly, HDAC class I cellular assay was conducted in K562 cell line in order to contrast with
extracellular inhibitory activity which utilized Hela extract as the enzyme source (mainly
contained class I HDACs: HDAC1 and HDAC2). As shown in Table 4, cellular class I HDAC

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activity of compound 4a only had a slightly enhancement compared with extracellular activity
(IC50 = 138 nM). But compound 4e displayed substantial increase in cellular class I HDAC
activity (IC50 = 64 nM) in contrast to extracellular activity (IC50 = 275 nM). These results
indicated that compounds 4a and 4e possess good cell permeability in K562 cell line and their
cellular enzyme inhibitory activities were consisted with their anti-proliferative activities.
Furthermore, the result of class IIa cellular assay indicated that class IIa HDACs were less
sensitive to compounds 4a and 4e as their IC50 value ranging in the micromolar concentration.
These results in celluar level as well as the HDAC isoforms selectivity results (Table 3) suggested
that compounds 4a and 4e might be potent class I HDAC inhibitors.
Table 4 HDAC class I and class IIa whole cell assay
Cpd.
HDACs class I cellular IC50
HDACs class IIa cellular IC50
(nM) a,c
(nM) a,b
108±13
64±8
5213±563
4a
4e
4208±469
Vorinostat
473±47
42275±2729
a Assays were performed in K562 cells; Values are average of at least two determinations, the SD
values are < 20% of the mean;
b Substrate: Boc-Lys (acetyl)-AMC;
c Substrate: Boc-Lys-(ε-trifluormethylacetyl)-AMC.
2.6 Apoptotic assay
It has been previously reported that HDACIs can promote cell apoptosis in vitro[25, 26]. Hence,
we chose 4a, 4e and Vorinostat (positive control) to perform flow cytometry analysis in K562 cells
and explore their abilities in inducing cancer cells apoptosis. Results in Fig.2 showed that
compounds 4a, 4e and Vorinostat could induce apoptosis in a dose-dependent manner. Moreover,
compound 4a and 4e exerted better pro-apoptosis activities than Vorinostat.

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Fig. 2 Inducing apoptosis of K562 cells by 4a, 4e and Vorinostat at 1µM and 2 µM after 24 h.
2.7 Molecular docking and molecular dynamics simulation
In order to explore the HDAC-inhibitor binding interactions, we further performed molecular
docking as well as molecular dynamics (MD) simulation. Although the biological functions of 11
zinc-dependent HDACs are not fully understood, HDAC1 has been widely recognized as an
important cancer relevant target[27]. Thus, HDAC1 and the most potent HDAC1 inhibitor,
compound 4a, were used in our computational study.
Firstly, compound 4a was docked into the active site of HDAC1 using Autodock Vina[28].
Then, the top-scored conformation was subjected to MD simulation. After energy minimization,
heat and equilibration, a 20ns standard MD simulation was performed. As shown in Fig. 3,
compound 4a remained in the HDAC1 active site during 20ns MD simulation and became stable
after 9ns MD simulation. Clustering analysis was performed using the last 10ns snapshots and
representative structure from the biggest cluster suggested the favorable binding mode for
compound 4a with HDAC1. The hydroxamic acid group of compound 4a chelated with zinc atom
properly and formed a series of hydrogen bond interactions with key residues (His140, His141,
Asp176, His178 and Tyr301).
Asp99
A
B
C
4a
Compound 4a
Protein backbone
Phe205
Phe150
4a
His178
Tyr303
His141
His140
d1
Cluster analysis
Zn2+
d1=2.14±0.04
d2=2.09±0.04
Zn2+
Asp176
d2
Asp264
t (ns)
Fig. 3 Predication of the binding mode of compound 4a in HDAC1. (a) RMSD values of
compound 4a (forest green) and HDAC1 protein backbone (blue) during 20ns MD simulation.
Snapshots in the last 10ns (grey) were used in clustering analysis. (b) The zinc chelating mode of
compound 4a (green). Average distances were measured using the last 10ns snapshots of MD
simulation. Active site of HDAC1 was shown as mesh. (c) Interactions between compound 4a
(green) and HDAC1 residues (white). Hydrogen bonds were shown as yellow dotted line.

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3. Conclusions
In our current study, we developed a novel series of quinoline-based N-hydroxycinnamamides
and N-hydroxybenzamides, showed their biological evaluation for Hela extract inhibitory
activities and anti-proliferative activities on cancer cells. Compound 4a exhibited higher Hela
extract inhibitory activity than Vorinostat. Besides, compounds 4a and 4e showed good anti-
proliferative activities with IC50 being 3-5 folds lower than Vorinostat in K562 and PC-3 cell lines.
Meanwhile, HDAC isoforms inhibitory assay and HDAC whole cell assay revealed compounds 4a
and 4e exhibited selective inhibition against class I HDACs. Moreover, flow-cytometry analysis
suggested that compounds 4a and 4e induced stronger apoptotic effects in K562 cells than that of
Vorinostat, which were in line with their anti-proliferative activities. With the inspiriting results
that compound 4a displayed class I HDACs inhibition and inducing apoptotic effect, it provided a
starting point for development of novel HDAC inhibitors targeted to class I HDACs.
4. Experimental section
4.1. Chemistry
Reagents and solvents used were of commercially available LR grade quality and without
further purification. All reactions were monitored by TLC on 0.25 mm silica gel plates
(60GF-254). UV light, chloride ferric and iodine vapor were used to visualize the spots. Melting
points were recorded by the RY-1G electrothermal melting point apparatus without correction. 1H
and 13C NMR spectra were recorded on a Brucker DRX spectrometer at 300 MHz or 400 MHz, d
in parts per million and J in hertz, using TMS as an internal standard. Significant 1H NMR data are
reported in the following order: multiplicity (s, singlet; d, doublet; t, triplet; m, multiplet) number
of protons. High-resolution mass spectra (HRMS) were conducted on an Agilent 6510 Quadrupole
Time-of-Flight LC/MS deliver. The purity of all targeted compounds was determined to
be >95.0% by HPLC. HPLC analysis was performed using an Diamonsil C18 (5µ, 250 mm×4.6
mm) and as a mobile phase gradient from 10% MeCN / 90%H2O (1‰ formic acid) for 5 min,
10% MeCN / 90%H2O (1‰ formic acid) to 90% MeCN / 10%H2O (1‰ formic acid) for 25 min,
90% MeCN / 10%H2O (1‰ formic acid) for 10 min, a flow rate of 1.0 mL/min.

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4.1.1. Methyl (E)-3-(4-(((quinolin-2-ylmethyl)amino)methyl)phenyl)acrylate (3a)
Aldehyde 1a (0.16g, 1mmol) and compound 2 (0.19g, 1mmol) was dissolved in MeOH (25ml)
and heated at 50 °C for 2 h. After cooling to 0 °C, NaBH4 (3 equiv) was added to the reaction
mixture, then the solution was stirred for 2 h at RT. The MeOH was evaporated under reduced
pressure and EtOAc was added. The EtOAc layer was washed with a saturated solution of
NaHCO3 (×3), brine solution (×3) and then dried and reduced to yield an oil (55%). The oil was
purified by column chromatography (DCM / MeOH 120:1). 1H NMR (400 MHz, CDCl3) δ 8.12 (d,
J = 8.4 Hz, 1H), 8.06 (d, J = 8.5 Hz, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.73 – 7.66 (m, 2H), 7.54 –
7.49 (m, 3H), 7.44 (t, J = 8.6 Hz, 3H), 6.43 (d, J = 16.0 Hz, 1H), 4.12 (s, 2H), 3.94 (s, 2H), 3.81
(s, 3H).
4.1.2. Methyl (E)-3-(4-((((6-chloroquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate(3b)
Using the synthetic method for 3a, the target compound was obtained as a yellow oil in 63%
yield. 1H NMR (400 MHz, CDCl3) δ 8.05 (d, J = 8.5 Hz, 1H), 8.00 (d, J = 9.0 Hz, 1H), 7.79 (d, J
= 2.3 Hz, 1H), 7.71 – 7.63 (m, 2H), 7.51 (d, J = 8.2 Hz, 2H), 7.48 – 7.42 (m, 3H), 6.42 (d, J =
16.0 Hz, 1H), 4.15 (s, 2H), 3.99 (s, 2H), 3.81 (s, 3H).
4.1.3. Methyl (E)-3-(4-((((6-bromoquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate (3c)
Using the synthetic method for 3a, the target compound was obtained as a yellow oil in 53%
yield. 1H NMR (400 MHz, CDCl3) δ 8.04 (d, J = 8.5 Hz, 1H), 7.97 (d, J = 2.1 Hz, 1H), 7.93 (d, J
= 8.9 Hz, 1H), 7.77 (dd, J = 9.0, 2.2 Hz, 1H), 7.68 (d, J = 16.0 Hz, 1H), 7.51 (d, J = 8.2 Hz, 2H),
7.48 – 7.43 (m, 3H), 6.43 (d, J = 16.0 Hz, 1H), 4.14 (s, 2H), 3.98 (s, 2H), 3.81 (s, 3H).
4.1.4. Methyl (E)-3-(4-((((6-methoxyquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate(3d)
Using the synthetic method for 3a, the target compound was obtained as a yellow oil in 39%
yield. 1H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 8.4 Hz, 1H), 7.95 (d, J = 9.2 Hz, 1H), 7.69 (d, J
= 16.0 Hz, 1H), 7.49 (t, J = 8.3 Hz, 2H), 7.42 (t, J = 8.0 Hz, 2H), 7.40 – 7.33 (m, 2H), 7.07 (d, J =
2.7 Hz, 1H), 6.42 (d, J = 16.0 Hz, 1H), 4.60 (s, 2H), 4.08 (s, 2H), 3.93 (s, 3H), 3.81 (s, 3H).
4.1.5. Methyl (E)-3-(4-((((8-chloroquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate (3e)
Using the synthetic method for 3a, the target compound was obtained as a yellow oil in 62%
1
yield. H NMR (400 MHz, CDCl3) δ 8.12 (d, J = 8.4 Hz, 1H), 7.82 (d, 1H), 7.71 (dd, J = 14.3,
12.2 Hz, 2H), 7.53 – 7.40 (m, 6H), 6.43 (d, J = 16.0 Hz, 1H), 4.16 (s, 2H), 3.96 (s, 2H), 3.80 (s,
3H).
4.1.6. Methyl (E)-3-(4-((((8-methoxyquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate(3f)
Using the synthetic method for 3a, the target compound was obtained as a yellow oil in 49%
yield. 1H NMR (400 MHz, CDCl3) δ 8.10 (d, J = 8.5 Hz, 1H), 7.69 (d, J = 16.0 Hz, 1H), 7.57 (d,
J = 8.4 Hz, 1H), 7.49 (d, J = 8.2 Hz, 2H), 7.44 – 7.40 (m, 3H), 7.38 (m, 1H), 7.05 (d, J = 7.5, 1.1
Hz, 1H), 6.42 (d, J = 16.0 Hz, 1H), 4.16 (s, 2H), 4.07 (s, 3H), 3.91 (s, 2H), 3.80 (s, 3H).
4.1.7. (E)-N-hydroxy-3-(4-(((quinolin-2-ylmethyl)amino)methyl)phenyl)acrylamide (4a)
Preparation of hydroxylamine in methanol:

ACCEPTED MANUSCRIPT
Solution A was obtained by hydroxylamine hydrochloride (4.67g, 67 mmol) dissolving in
methanol (24 ml). Solution B was acquired by potassium hydroxide (6.60 g, 100 mmol) diluting in
methanol (14 ml). Next, solution B was added dropwise to the solution A at 0 °C. The reaction
mixture was stirred for 30 min at 0 °C and filtered to give the solution C.
Compound 3a (0.33g) was dissolved in the solution C (12 mL), and the reaction was stirred at
RT for 4 h. Then the mixture was neutralized with 1 M HCl to Ph 7. White precipitate obtained
was filtered and recrystallization to obtain a white solid (0.15g, 45%). mp：202-203 °C. 1H NMR
(400 MHz, DMSO) δ 10.90 (s, 1H), 9.84 (s, 1H), 9.10 (s, 1H), 8.46 (d, J = 8.5 Hz, 1H), 8.08 (d, J
= 8.4 Hz, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.84 (dd, J = 11.2, 4.1 Hz, 1H), 7.71 – 7.61 (m, 6H), 7.49
(d, J = 15.8 Hz, 1H), 6.55 (d, J = 15.8 Hz, 1H), 4.52 (s, 2H), 4.34 (s, 2H). 13C NMR (101 MHz,
DMSO) δ 163.02, 153.45, 147.00, 137.89, 137.68, 135.95, 133.52, 131.41, 130.68, 128.87, 128.57,
128.04, 127.69, 127.45, 121.20, 120.71, 50.51, 50.25. HRMS (AP-ESI) m/z Calcd for C20H19N3O2
[M+H]+ 334.155, found: 334.1560.
4.1.8.(E)-3-(4-((((6-chloroquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacrylamide (4b)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 29%
yield. mp：202-205 °C. 1H NMR (400 MHz, DMSO) δ 10.99 (s, 1H), 10.02 (s, 2H), 9.10 (s, 1H),
8.44 (d, J = 8.6 Hz, 1H), 8.19 (d, J = 2.4 Hz, 1H), 8.08 (d, J = 9.0 Hz, 1H), 7.84 (dd, J = 9.0, 2.4
Hz, 1H), 7.75 (d, J = 8.6 Hz, 1H), 7.68 (d, J = 8.2 Hz, 2H), 7.62 (d, J = 8.2 Hz, 2H), 7.48 (d, J =
15.8 Hz, 1H), 6.61 (d, J = 15.9 Hz, 1H), 4.49 (s, 2H), 4.32 (s, 2H). 13C NMR (101 MHz, DMSO)
δ 163.06, 155.21, 152.97, 138.88, 138.02, 137.40, 135.72, 134.89, 131.16, 128.74, 128.05, 127.67,
121.62, 120.43, 119.94, 109.56, 56.15, 51.22, 50.73. HRMS (AP-ESI) m/z Calcd for
C20H18ClN3O2 [M+H]+ 368.116, found: 368.1171.
4.1.9.(E)-3-(4-((((6-bromoquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacrylamide (4c)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 31%
yield. mp：144-146 °C. 1H NMR (400 MHz, DMSO) δ 10.86 (s, 1H), 9.07 (s, 1H), 8.43 (d, J = 8.6
Hz, 1H), 8.35 (d, J = 2.0 Hz, 1H), 8.01 (d, J = 9.0 Hz, 1H), 7.95 (dd, J = 9.0, 2.1 Hz, 1H), 7.69 (d,
J = 8.6 Hz, 1H), 7.61 (d, J = 12.2 Hz, 4H), 7.48 (d, J = 15.9 Hz, 1H), 6.53 (d, J = 15.8 Hz, 1H),
4.47 (s, 2H), 4.29 (s, 2H). 13C NMR (101 MHz, DMSO) δ 163.00, 154.80, 145.64, 143.66, 137.95,
136.84, 135.81, 133.66, 131.23, 131.15, 131.08, 130.55, 128.98, 128.78, 128.04, 122.17, 120.56,
120.26, 50.77, 50.44. HRMS (AP-ESI) m/z Calcd for C20H18BrN3O2 [M+H]+ 412.0655, found:
412.0655.
4.1.10 (E)-N-hydroxy-3-(4-((((6-methoxyquinolin-2-yl)methyl)amino)methyl)phenyl)acrylamide
(4d)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 34%
yield. mp：148-151 °C. 1H NMR (400 MHz, DMSO) δ 10.90 (s, 1H), 9.08 (s, 1H), 8.31 (d, J = 8.5
Hz, 1H), 7.94 (d, J = 9.1 Hz, 1H), 7.65 – 7.55 (m, 5H), 7.51 – 7.39 (m, 3H), 6.55 (d, J = 15.8 Hz,
1H), 4.31 (s, 2H), 4.18 (s, 2H), 3.91 (s, 3H). 13C NMR (101 MHz, DMSO) δ 163.03, 157.84,
152.64, 143.14, 138.01, 136.26, 135.78, 135.41, 130.79, 130.33, 128.80, 127.99, 122.82, 121.46,
120.30, 106.30, 56.03, 51.52, 50.82. HRMS (AP-ESI) m/z Calcd for C21H21N3O3 [M+H]+
364.1656, found: 364.1656.

ACCEPTED MANUSCRIPT
4.1.11(E)-3-(4-((((8-chloroquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacrylamide (4e)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 26%
yield. mp：159-161 °C. 1H NMR (400 MHz, DMSO) δ 10.87 (s, 1H), 9.07 (s, 1H), 8.54 (d, J = 8.5
Hz, 1H), 8.06 – 8.00 (m, 2H), 7.80 (d, J = 8.5 Hz, 1H), 7.66 – 7.59 (m, 5H), 7.48 (d, J = 15.8 Hz,
1H), 6.55 (d, J = 15.8 Hz, 1H), 4.52 (s, 2H), 4.38 (s, 2H). 13C NMR (101 MHz, DMSO) δ 163.04,
155.42, 142.95, 138.27, 137.95, 135.74, 134.57, 132.32, 131.12, 130.67, 129.12, 128.80, 128.03,
127.64, 122.27, 120.50, 50.98, 50.65. HRMS (AP-ESI) m/z Calcd for C20H18ClN3O2 [M+H]+
368.116, found: 368.1161.
4.1.12 (E)-N-hydroxy-3-(4-((((8-methoxyquinolin-2-yl)methyl)amino)methyl)phenyl)acrylamide
(4f)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 29%
yield. mp：151-154 °C. 1H NMR (400 MHz, DMSO) δ 10.82 (s, 1H), 9.07 (s, 1H), 8.39 (d, J = 8.5
Hz, 1H), 7.67 – 7.59 (m, 5H), 7.56 (d, J = 6.7 Hz, 2H), 7.52 (d, J = 7.7 Hz, 1H), 7.46 (d, J = 9.8
Hz, 1H), 7.29 (dd, J = 6.8, 2.2 Hz, 1H), 6.53 (d, J = 15.9 Hz, 1H), 4.45 (s, 2H), 4.28 (s, 2H), 4.02
(s, 3H). 13C NMR (101 MHz, DMSO) δ 163.06, 155.21, 152.97, 138.88, 138.02, 137.40, 135.72,
134.89, 131.16, 128.74, 128.05, 127.67, 121.62, 120.43, 119.94, 109.56, 56.15, 51.22, 50.73.
HRMS (AP-ESI) m/z Calcd for C21H21N3O3 [M+H]+ 364.1656, found: 364.1656.
4.1.13
(E)-3-(3-((((6-bromoquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacrylamide
(4g)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 36%
yield. mp：176-178 °C. 1H NMR (400 MHz, DMSO) δ 10.75 (s, 1H), 9.06 (s, 1H), 8.32 (d, J = 8.5
Hz, 1H), 8.26 (d, J = 2.1 Hz, 1H), 7.91 (d, J = 9.0 Hz, 1H), 7.84 (dd, J = 8.9, 2.2 Hz, 1H), 7.73 (d,
J = 8.5 Hz, 1H), 7.58 (s, 1H), 7.47 – 7.32 (m, 4H), 6.47 (d, J = 15.8 Hz, 1H), 3.97 (s, 2H), 3.78 (s,
2H). 13C NMR (101 MHz, DMSO) δ 163.17, 162.42, 146.06, 141.85, 138.85, 136.00, 135.18,
132.90, 131.13, 130.26, 129.70, 129.23, 128.75, 127.27, 126.59, 121.95, 119.46, 119.22, 54.95,
52.78. HRMS (AP-ESI) m/z Calcd for C20H18BrN3O2 [M+H]+ 412.0655, found: 412.0667.
4.1.14 Methyl (E)-3-(4-((benzyl(quinolin-2-ylmethyl)amino)methyl)phenyl)acrylate (5a)
Compound 4a(0.60g, 1.7mmol), Benzyl bromide (0.29g, 1.7mmol) and DIEA(3 equiv) was
dissolved in DMSO (10 ml) and stirred for 4h at 50 °C. After cooled down, the mixture was
poured into 100 ml H2O and extracted with DCM. The solution was evaporated under vacuum to
yield an oil (48%). The oil was purified by column chromatography (DCM/ MeOH 90:1).
1H NMR (400 MHz, DMSO) δ 8.36 (d, J = 8.5 Hz, 1H), 8.01 (d, J = 8.4 Hz, 1H), 7.95 (d, J =
7.5 Hz, 1H), 7.77 – 7.64 (m, 5H), 7.58 (t, 1H), 7.49 (d, J = 8.1 Hz, 2H), 7.44 (d, J = 7.2 Hz, 2H),
7.37 (t, J = 7.5 Hz, 2H), 7.27 (t, J = 7.3 Hz, 1H), 6.63 (d, J = 16.1 Hz, 1H), 3.82 (s, 2H), 3.75 (s,
3H), 3.62 (s, 2H), 3.61 (s, 2H).
4.1.15
Methyl (E)-3-(4-((benzyl((6-chloroquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate
(5b)

ACCEPTED MANUSCRIPT
Using the synthetic method for 5a, the target compound was obtained as a light yellow oil in
56% yield. 1H NMR (400 MHz, CDCl3) δ 8.02 (d, J = 8.5 Hz, 1H), 7.96 (d, J = 9.0 Hz, 1H), 7.77
– 7.71 (m, 2H), 7.67 (d, J = 16.0 Hz, 1H), 7.60 (dd, J = 9.0, 2.3 Hz, 1H), 7.47 (d, J = 8.2 Hz, 2H),
7.45 – 7.38 (m, 4H), 7.33 (t, J = 7.5 Hz, 2H), 7.27 – 7.21 (m, 1H), 6.41 (d, J = 16.0 Hz, 1H), 3.87
(s, 2H), 3.79 (s, 3H), 3.64 (s, 2H), 3.64 (s, 2H).
4.1.16 Methyl (E)-3-(4-((benzyl((6-bromoquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate
(5c)
Using the synthetic method for 5a, the target compound was obtained as a light yellow oil in
59% yield. 1H NMR (400 MHz, DMSO) δ 8.36 – 8.30 (m, 2H), 8.22 (d, J = 2.2 Hz, 1H), 7.92 (d,
J = 9.0 Hz, 1H), 7.83 (dd, J = 9.0, 2.2 Hz, 1H), 7.76 (d, J = 8.5 Hz, 1H), 7.72 – 7.62 (m, 3H),
7.47 (d, J = 8.1 Hz, 2H), 7.43 (d, J = 7.2 Hz, 2H), 7.36 (t, J = 7.5 Hz, 2H), 7.26 (t, J = 7.3 Hz,
1H), 6.61 (d, J = 16.1 Hz, 1H), 3.80 (s, 2H), 3.75 (s, 3H), 3.61 (s, 2H), 3.60 (s, 2H).
4.1.17 Methyl (E)-3-(4-((benzyl((6-methoxyquinolin-2-yl)methyl)amino)methyl)phenyl)acrylate
(5d)
Using the synthetic method for 5a, the target compound was obtained as a light yellow oil in
47% yield. 1H NMR (400 MHz, CDCl3) δ 8.02 (d, J = 8.5 Hz, 1H), 7.94 (d, J = 9.2 Hz, 1H), 7.71
– 7.63 (m, 2H), 7.47 (d, J = 8.3 Hz, 2H), 7.45 – 7.37 (m, 4H), 7.36 – 7.30 (m, 3H), 7.26 – 7.21 (m,
1H), 7.05 (d, J = 2.7 Hz, 1H), 6.41 (d, J = 16.0 Hz, 1H), 3.90 (s, 3H), 3.86 (s, 2H), 3.79 (s, 3H),
3.63 (s, 4H).
4.1.18 Methyl (E)-3-(4-((((6-bromoquinolin-2-yl)methyl)(hexyl)amino)methyl)phenyl)acrylate
(5e)
Using the synthetic method for 5a, the target compound was obtained as a light yellow oil in
46% yield. 1H NMR (400 MHz, DMSO) δ 8.29 (d, J = 8.6 Hz, 1H), 8.20 (d, J = 2.0 Hz, 1H), 7.88
(d, J = 9.0 Hz, 1H), 7.80 (dd, J = 8.9, 2.1 Hz, 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.66 – 7.59 (m, 3H),
7.39 (d, J = 8.1 Hz, 2H), 6.58 (d, J = 16.0 Hz, 1H), 3.77 (s, 2H), 3.71 (s, 3H), 3.59 (s, 2H), 2.37 (t,
J = 7.1 Hz, 2H), 1.47 – 1.37 (m, 2H), 1.19 – 1.08 (m, 4H), 1.08 – 0.98 (m, 2H), 0.73 (t, J = 7.2 Hz,
3H).
4.1.19 (E)-3-(4-((benzyl(quinolin-2-ylmethyl)amino)methyl)phenyl)-N-hydroxyacrylamide (6a)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 17%
yield. mp：100-103 °C. 1H NMR (400 MHz, DMSO) δ 10.73 (s, 1H), 9.01 (s, 1H), 8.36 (d, J = 8.4
Hz, 1H), 7.97 (d, J = 8.5 Hz, 1H), 7.94 (d, J = 7.9 Hz, 1H), 7.76 – 7.69 (m, 2H), 7.60 – 7.51 (m,
3H), 7.49 – 7.40 (m, 5H), 7.35 (t, J = 7.5 Hz, 2H), 7.26 (t, J = 7.3 Hz, 1H), 6.43 (d, J = 15.9 Hz,
1H), 3.81 (s, 2H), 3.61 (s, 2H), 3.60 (s, 2H). 13C NMR (101 MHz, CDCl3) δ 160.69, 147.02,
141.03, 138.82, 136.86, 133.39, 129.71, 129.17, 128.83, 128.32, 127.86, 127.58, 127.39, 127.12,
126.32, 120.90, 60.08, 58.41, 57.97. HRMS (AP-ESI) m/z Calcd for C27H25N3O2 [M+H]+
424.2020, found: 424.2029.

ACCEPTED MANUSCRIPT
4.1.20
(E)-3-(4-((benzyl((6-chloroquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacrylamide
(6b)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 26%
yield. mp：110-112 °C. 1H NMR (400 MHz, DMSO) δ 8.34 (d, J = 8.5 Hz, 1H), 8.09 (d, J = 2.3
Hz, 1H), 7.99 (d, J = 9.0 Hz, 1H), 7.77 (d, J = 8.6 Hz, 1H), 7.73 (dd, J = 9.0, 2.4 Hz, 1H), 7.50 (d,
J = 8.0 Hz, 2H), 7.43 (d, J = 8.6 Hz, 4H), 7.37 – 7.31 (m, 3H), 7.28 – 7.23 (m, 1H), 6.41 (d, J =
15.8 Hz, 1H), 3.80 (s, 2H), 3.59 (s, 4H). 13C NMR (101 MHz, DMSO) δ 163.26, 161.24, 145.82,
140.81, 139.07, 138.57, 136.34, 134.16, 131.09, 130.94, 130.40, 129.64, 129.13, 128.78, 128.23,
127.98, 127.54, 126.97, 122.16, 119.16, 59.88, 57.88, 57.53. HRMS (AP-ESI) m/z Calcd for
C27H24ClN3O2 [M+H]+ 458.1630, found: 458.1637.
4.1.21
(E)-3-(4-((benzyl((6-bromoquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacrylamide
(6c)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 31%
yield. mp：117-119 °C. 1H NMR (400 MHz, DMSO) δ 10.73 (s, 1H), 9.04 (s, 1H), 8.35 (d, J = 8.5
Hz, 1H), 8.25 (d, J = 1.7 Hz, 1H), 7.92 (d, J = 9.0 Hz, 1H), 7.84 (dd, J = 9.0, 2.0 Hz, 1H), 7.77 (d,
J = 8.5 Hz, 1H), 7.54 (d, J = 7.9 Hz, 2H), 7.49 – 7.40 (m, 5H), 7.35 (t, J = 7.5 Hz, 2H), 7.25 (t, J
= 7.2 Hz, 1H), 6.43 (d, J = 15.9 Hz, 1H), 3.79 (s, 2H), 3.61 (s, 2H), 3.60 (s, 2H).13C NMR (101
MHz, CDCl3) δ 165.33, 161.19, 145.64, 140.93, 138.66, 135.68, 133.40, 132.99, 130.18, 129.57,
129.20, 128.83, 128.44, 128.36, 127.89, 127.20, 121.73, 120.05, 116.15, 60.01, 58.45, 58.01.
HRMS (AP-ESI) m/z Calcd for C27H24BrN3O2 [M+H]+ 502.1125, found: 502.1123.
4.1.22(E)-3-(4-((benzyl((6-methoxyquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacryla
mide (6d)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 29%
yield. mp：94-96 °C. 1H NMR (400 MHz, DMSO) δ 10.75 (s, 1H), 9.02 (s, 1H), 8.25 (d, J = 8.5
Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.54 (d, J = 8.0 Hz, 2H), 7.44 (dd, J =
13.0, 7.5 Hz, 5H), 7.40 – 7.31 (m, 4H), 7.26 (t, J = 7.3 Hz, 1H), 6.44 (d, J = 15.8 Hz, 1H), 3.88 (s,
3H), 3.76 (s, 2H), 3.59 (d, J = 2.7 Hz, 4H). 13C NMR (101 MHz, CDCl3) δ 164.69, 158.03, 157.55,
143.12, 141.06, 140.47, 139.00, 135.61, 133.56, 129.87, 129.14, 128.83, 128.36, 128.30, 127.81,
127.06, 122.06, 121.23, 116.78, 105.30, 60.05, 58.35, 57.95, 55.53. HRMS (AP-ESI) m/z Calcd
for C28H27N3O3 [M+H]+ 454.2125, found: 454.2122.
4.1.23
(E)-3-(4-((((6-bromoquinolin-2-yl)methyl)(hexyl)amino)methyl)phenyl)-N-hydroxyacrylamide
(6e)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 37%
yield. mp：82-84 °C. 1H NMR (400 MHz, DMSO) δ 10.75 (s, 1H), 9.03 (s, 1H), 8.33 (d, J = 8.6
Hz, 1H), 8.24 (d, J = 2.1 Hz, 1H), 7.91 (d, J = 9.0 Hz, 1H), 7.84 (dd, J = 9.0, 2.2 Hz, 1H), 7.72 (d,
J = 8.5 Hz, 1H), 7.52 (d, J = 8.0 Hz, 2H), 7.47 – 7.38 (m, 3H), 6.44 (d, J = 15.8 Hz, 1H), 3.81 (s,
2H), 3.62 (s, 2H), 2.42 (t, J = 7.0 Hz, 2H), 1.52 – 1.40 (m, 2H), 1.18 (dt, J = 14.3, 7.2 Hz, 4H),
1.12 – 1.03 (m, 2H), 0.76 (t, J = 7.2 Hz, 3H). 13C NMR (101 MHz, CDCl3) δ 161.04, 148.30,

ACCEPTED MANUSCRIPT
145.76, 140.70, 135.51, 133.60, 132.89, 130.35, 129.56, 129.32, 128.46, 127.93, 121.91, 120.34,
119.98, 116.73, 60.37, 58.43, 54.20, 31.59, 29.70, 26.88, 22.57, 14.02. HRMS (AP-ESI) m/z
Calcd for C26H30BrN3O2 [M+H]+ 496.1597, found: 496.1604.
4.1.24
(E)-3-(3-((benzyl((6-bromoquinolin-2-yl)methyl)amino)methyl)phenyl)-N-hydroxyacrylamide
(6f)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 33%
yield. mp：82-84 °C. 1H NMR (400 MHz, DMSO) δ 10.82 (s, 1H), 9.10 (s, 1H), 8.33 (d, J = 9.1
Hz, 1H), 8.22 (d, J = 2.1 Hz, 1H), 7.90 (d, J = 9.0 Hz, 1H), 7.82 (dd, J = 9.0, 2.2 Hz, 1H), 7.77 (d,
J = 8.6 Hz, 1H), 7.60 (s, 1H), 7.48(d, J = 15.8 Hz, 1H), 7.46 – 7.31 (m, 7H), 7.25 (t, J = 7.3 Hz,
1H), 6.49 (d, J = 15.8 Hz, 1H), 3.79 (s, 2H), 3.59 (s, 4H). 13C NMR (101 MHz, CDCl3) δ 165.26,
161.31, 145.42, 141.14, 139.45, 138.70, 135.73, 134.54, 132.97, 130.24, 129.94, 129.55, 128.85,
128.68, 128.40, 128.35, 127.17, 126.74, 121.80, 120.03, 116.93, 60.01, 58.63, 58.19.
HRMS (AP-ESI) m/z Calcd for C27H24BrN3O2 [M+H]+ 502.1125, found: 502.1135.
4.1.25
(E)-3-(3-((((6-bromoquinolin-2-yl)methyl)(hexyl)amino)methyl)phenyl)-N-hydroxyacrylamide
(6g)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 26%
yield. mp：76-78 °C. 1H NMR (400 MHz, DMSO) δ 10.75 (s, 1H), 9.11 (s, 1H), 8.33 (d, J = 8.5
Hz, 1H), 8.24 (d, J = 1.7 Hz, 1H), 7.91 (d, J = 9.0 Hz, 1H), 7.84 (dd, J = 8.9, 2.0 Hz, 1H), 7.73 (d,
J = 8.5 Hz, 1H), 7.56 (s, 1H), 7.51 – 7.31 (m, 4H), 6.48 (d, J = 15.8 Hz, 1H), 3.82 (s, 2H), 3.63 (s,
2H), 2.43 (t, J = 6.9 Hz, 2H), 1.51 – 1.44 (m, 2H), 1.22 – 1.14 (m, 4H), 1.11 – 1.04 (m, 2H), 0.76
(t, J = 9.4, 5.0 Hz, 3H). 13C NMR (101 MHz, CDCl3) δ 164.82, 161.37, 145.63, 140.98, 139.50,
135.60, 134.64, 132.94, 130.41, 130.14, 129.58, 128.64, 128.46, 127.66, 126.80, 121.99, 120.00,
116.84, 60.46, 58.61, 54.38, 31.61, 29.72, 26.91, 22.59, 14.03. HRMS (AP-ESI) m/z Calcd for
C26H30BrN3O2 [M+H]+ 496.1594, found: 496.1604.
4.1.26 N-(quinolin-2-ylmethyl)aniline (7a)
Aldehyde 1a (0.16g, 1 mmol) was dissolved in DCM (10 ml) and aniline (0.11g, 1.2 mmol) was
added dropwise. The reaction was stirred at RT for overnight, and a crude product was obtained
after evaporation of the solvent. Then MeOH and NaBH4 (0.113 g, 3 mmol) was added, and
reaction mixture was stirred 2h at RT. The mixture was concentrated under reduced pressure and
added saturated NaCO3 solution. The aqueous phase was extracted with EtOAc, dried over
anhydrous MgSO4 and evaporated to give crude residue. The crude product was purified by
column chromatography (petroleum ether/EtOAc 30: 1). mp：56-58 °C.1H NMR (400 MHz,
CDCl3) δ 8.12 (t, 2H), 7.82 (d, J = 8.3 Hz, 1H), 7.73 (t, 1H), 7.54 (t, 1H), 7.46 (d, 1H), 7.21 (t, J =
7.8 Hz, 2H), 6.74 (t, J = 8.7 Hz, 3H), 5.14 (s, 1H), 4.64 (d, J = 5.2 Hz, 2H).
4.1.27 4-Bromo-N-(quinolin-2-ylmethyl)aniline (7b)
Using the synthetic method for 7a, the target compound was obtained as a white solid in 66%
yield. mp：128-131 °C. 1H NMR (400 MHz, CDCl3) δ 8.11 (dd, J = 11.3, 8.6 Hz, 2H), 7.82 (d, J =
8.1 Hz, 1H), 7.74 (t, J = 7.6 Hz, 1H), 7.54 (t, J = 7.5 Hz, 1H), 7.41 (d, J = 8.4 Hz, 1H), 7.27 (d, J
= 8.8 Hz, 2H), 6.63 (d, J = 8.7 Hz, 2H), 5.27 (s, 1H), 4.59 (d, J = 5.1 Hz, 2H).

ACCEPTED MANUSCRIPT
4.1.28 4-Methoxy-N-(quinolin-2-ylmethyl)aniline (7c)
Using the synthetic method for 7a, the target compound was obtained as a light yellow solid in
61% yield. mp：138-141 °C. 1H NMR (400 MHz, CDCl3) δ 8.11 (t, J = 7.6 Hz, 2H), 7.81 (d, J =
7.9 Hz, 1H), 7.73 (t, J = 7.8 Hz, 1H), 7.54 (d, J = 7.5 Hz, 1H), 7.46 (d, J = 8.5 Hz, 1H), 6.80 (d, J
= 8.9 Hz, 2H), 6.71 (d, J = 8.9 Hz, 2H), 4.60 (s, 2H), 3.75 (s, 3H).
4.1.29 N-((8-chloroquinolin-2-yl)methyl)-4-methoxyaniline (7d)
Using the synthetic method for 7a, the target compound was obtained as a white solid in 58%
yield. 1H NMR (400 MHz, CDCl3) δ 8.14 (d, J = 8.5 Hz, 1H), 7.85 (d, J = 7.5, 1.1 Hz, 1H), 7.75
(d, 1H), 7.51 (d, J = 8.5 Hz, 1H), 7.47 (t, J = 7.8 Hz, 1H), 6.81 (d, J = 1.8 Hz, 4H), 4.68 (s, 2H),
3.75 (s, 3H).
4.1.30 N-((6-bromoquinolin-2-yl)methyl)aniline (7e)
Using the synthetic method for 7a, the target compound was obtained as a white solid in 41%
yield. mp：192-193°C.1H NMR (400 MHz, CDCl3) δ 8.06 – 7.95 (m, 3H), 7.79 (dd, J = 8.9, 2.2
Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 7.20 (dd, J = 8.6, 7.3 Hz, 2H), 6.73 (d, J = 7.7 Hz, 3H), 4.62 (s,
2H).
4.1.31 N-((8-methoxyquinolin-2-yl)methyl)aniline(7f)
Using the synthetic method for 7a, the target compound was obtained as a brown solid in 46%
yield. 1H NMR (400 MHz, CDCl3) δ 8.09 (d, J = 8.5 Hz, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.44 (d, J
= 7.8 Hz, 1H), 7.39 (d, J = 7.3 Hz, 1H), 7.21 – 7.14 (m, 2H), 7.09 (d, J = 7.5 Hz, 1H), 6.75 – 6.66
(m, 3H), 4.73 (s, 2H), 4.12 (s, 3H).
4.1.32 N-((6-methoxyquinolin-2-yl)methyl)aniline (7g)
Using the synthetic method for 7a, the target compound was obtained as a pale yellow solid in
49% yield. 1H NMR (400 MHz, CDCl3) δ 8.04 (d, J = 8.4 Hz, 2H), 7.43 (d, J = 8.5 Hz, 1H), 7.39
(dd, J = 9.2, 2.8 Hz, 1H), 7.21 (d, J = 7.1 Hz, 1H), 7.18 (d, J = 7.4 Hz, 1H), 7.08 (d, J = 2.8 Hz,
1H), 6.73 (m, 3H), 4.62 (s, 2H), 3.94 (s, 3H).
4.1.33 Methyl (E)-3-(4-(phenyl(quinolin-2-ylmethyl)carbamoyl)phenyl)acrylate (10a)
Compound 9a (0.82 g, 4 mmol) was dissolved in 20 mL DCM and one drop DMF was added.
Then oxalyl chloride (1 mL, 12 mmol) was added dropwise and the reaction was stirred at RT for
1h. After the mixture was evaropated, the residue was dissolved in DCM and again concentrated
in vacuo to offer quantitatively the crude chloride. A solution of chloride in DCM (20 mL) was
suspended with NaHCO3 (2.7 g) and compound 7a (0.47g, 2mmol) was added dropwise with
stirring. After the addition, the reaction was stirred for 4 h. Then the mixture was washed with
1M citric acid(×3), saturate solution of NaHCO3(×3) and brine solution(×3). The solution was
concentrated in vacuo, then purified via chlomatography on silica (petroleum ether / EtOAc=3:1)
to give a white solid as the target compound（0.5g，50%）. mp：128-131 °C. 1H NMR (400 MHz,
CDCl3) δ 8.16 (d, J = 8.5 Hz, 1H), 8.03 (d, J = 8.4 Hz, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.69 (t, J =
7.7 Hz, 1H), 7.61 (d, J = 3.8 Hz, 1H), 7.57 (d, J = 11.4 Hz, 1H), 7.52 (t, J = 7.5 Hz, 1H), 7.44 (d,
J = 8.2 Hz, 2H), 7.35 (d, J = 8.2 Hz, 2H), 7.18 – 7.08 (m, 5H), 6.39 (d, J = 16.0 Hz, 1H), 5.45 (s,

ACCEPTED MANUSCRIPT
2H), 3.79 (s, 3H).
4.1.34 Methyl (E)-3-(4-((4-bromophenyl)(quinolin-2-ylmethyl)carbamoyl)phenyl)acrylate (10b)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 33%
yield. mp：178-180 °C. 1H NMR (400 MHz, CDCl3) δ 8.16 (d, J = 8.5 Hz, 1H), 8.02 (d, J = 8.4
Hz, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.73 – 7.68 (m, 1H), 7.62 (s, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.52
(d, 1H), 7.44 (d, J = 8.3 Hz, 2H), 7.38 (d, J = 8.3 Hz, 2H), 7.28 (d, J = 8.8 Hz, 2H), 7.03 (d, J =
8.7 Hz, 2H), 6.41 (d, J = 16.0 Hz, 1H), 5.40 (s, 2H), 3.80 (s, 3H).
4.1.35 Methyl (E)-3-(4-((4-methoxyphenyl)(quinolin-2-ylmethyl)carbamoyl)phenyl)acrylate
(10c)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 44%
yield. mp：158-160 °C. 1H NMR (400 MHz, CDCl3) δ 8.16 (d, J = 8.5 Hz, 1H), 8.03 (d, J = 8.4
Hz, 1H), 7.81 (d, J = 8.2 Hz, 1H), 7.69 (t, J = 7.1 Hz, 1H), 7.59 (d, J = 16.1 Hz, 2H), 7.52 (t, J =
7.1 Hz, 1H), 7.43 (d, J = 7.6 Hz, 2H), 7.35 (d, J = 8.2 Hz, 2H), 7.02 (d, J = 8.3 Hz, 2H), 6.65 (d, J
= 8.7 Hz, 2H), 6.39 (d, J = 16.0 Hz, 1H), 5.41 (s, 2H), 3.79 (s, 3H), 3.69 (s, 3H).
4.1.36
Methyl (E)-3-(4-(((8-chloroquinolin-2-yl)methyl)(4-methoxyphenyl)carbamoyl)phenyl)acrylate
(10d)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 62%
yield. mp：170-172 °C. 1H NMR (400 MHz, CDCl3) δ 8.21 – 8.15 (m, 1H), 7.83 (d, J = 7.5 Hz,
1H), 7.74 (d, J = 8.1 Hz, 1H), 7.59 (d, J = 16.1 Hz, 2H), 7.48 – 7.41 (m, 3H), 7.34 (t, 4H), 6.71 (d,
J = 8.7 Hz, 2H), 6.39 (d, J = 16.0 Hz, 1H), 5.44 (s, 2H), 3.79 (s, 3H), 3.72 (s, 3H).
4.1.37 Methyl (E)-3-(4-(((6-bromoquinolin-2-yl)methyl)(phenyl)carbamoyl)phenyl)acrylate
(10e)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 57%
yield. mp：175-177 °C. 1H NMR (400 MHz, CDCl3) δ 8.08 (d, J = 8.5 Hz, 1H), 8.00 – 7.87 (m,
2H), 7.75 (dd, J = 8.9, 2.1 Hz, 1H), 7.65 – 7.53 (m, 2H), 7.43 (d, J = 8.3 Hz, 2H), 7.34 (d, J = 8.3
Hz, 2H), 7.20 – 7.06 (m, 5H), 6.38 (d, J = 16.0 Hz, 1H), 5.43 (s, 2H), 3.78 (s, 3H).
4.1.38 Methyl (E)-3-(4-(((8-methoxyquinolin-2-yl)methyl)(phenyl)carbamoyl)phenyl)acrylate
(10f)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 59%
yield. mp：162-165 °C. 1H NMR (400 MHz, CDCl3) δ 8.18 (t, J = 7.6 Hz, 1H), 7.67 (d, J = 8.5 Hz,
1H), 7.58 (d, J = 16.0 Hz, 1H), 7.46 (d, 1H), 7.45 – 7.42 (m, 2H), 7.40 (d, J = 7.4 Hz, 1H), 7.35 (d,
J = 8.3 Hz, 2H), 7.17 – 7.05 (m, 6H), 6.39 (d, J = 16.0 Hz, 1H), 5.61 (s, 2H), 4.07 (s, 3H), 3.79 (s,
3H).
4.1.39 Methyl (E)-3-(4-(2-oxo-2-(phenyl(quinolin-2-ylmethyl)amino)ethyl)phenyl)acrylate
(10g)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 49%

ACCEPTED MANUSCRIPT
yield. mp：168-170 °C. 1H NMR (400 MHz, CDCl3) δ 8.12 (d, J = 8.1 Hz, 1H), 7.98 (s, 1H), 7.79
(d, J = 8.0 Hz, 1H), 7.73 – 7.63 (m, 2H), 7.55 – 7.46 (m, 2H), 7.42 (d, J = 8.1 Hz, 2H), 7.34 –
7.28 (m, 3H), 7.15 (d, J = 7.7 Hz, 4H), 6.41 (d, J = 16.0 Hz, 1H), 5.23 (s, 2H), 3.81 (s, 3H), 3.59
(s, 2H).
4.1.40
Methyl(E)-3-(4-(2-(((6-bromoquinolin-2-yl)methyl)(phenyl)amino)-2-oxoethyl)phenyl)acrylate
(10h)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 44%
yield. mp：162-164 °C.1H NMR (400 MHz, CDCl3) δ 8.05 (d, J = 8.4 Hz, 1H), 7.96 (d, J = 1.8 Hz,
1H), 7.87 (s, 1H), 7.75 (d, J = 8.9 Hz, 1H), 7.67 (d, J = 16.0 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H),
7.42 (d, J = 8.1 Hz, 2H), 7.37 – 7.29 (m, 3H), 7.20 – 7.08 (m, 4H), 6.41 (d, J = 16.0 Hz, 1H), 5.21
(s, 2H), 3.81 (s, 3H), 3.58 (s, 2H).
4.1.41
Methyl(E)-3-(4-(2-(((6-methoxyquinolin-2-yl)methyl)(phenyl)amino)-2-oxoethyl)phenyl)acrylate
(10i)
Using the synthetic method for 10a, the target compound was obtained as a white solid in 55%
yield. mp：172-174 °C. 1H NMR (400 MHz, CDCl3) δ 8.03 (d, J = 7.2 Hz, 1H), 7.88 (s, 1H), 7.67
(d, J = 16.0 Hz, 1H), 7.47 (d, J = 8.4 Hz, 1H), 7.42 (d, J = 8.1 Hz, 2H), 7.37 – 7.27 (m, 4H), 7.18
– 7.04 (m, 5H), 6.41 (d, J = 16.0 Hz, 1H), 5.21 (s, 2H), 3.92 (s, 3H), 3.81 (s, 3H), 3.58 (s, 2H).
4.1.42
(E)-4-(3-(hydroxyamino)-3-oxoprop-1-en-1-yl)-N-phenyl-N-(quinolin-2-ylmethyl)benzamide
(11a)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 41%
yield. mp：176-180 °C. 1H NMR (300 MHz, DMSO) δ 10.75 (s, 1H), 9.05 (s, 1H), 8.36 (d, J = 8.5
Hz, 1H), 7.96 (dd, J = 8.1, 3.0 Hz, 2H), 7.78 – 7.70 (m, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.59 (dd, J
= 11.0, 3.9 Hz, 1H), 7.48 – 7.39 (m, 4H), 7.35 (d, 1H), 7.22 (dt, J = 15.2, 7.6 Hz, 4H), 7.09 (t, J =
7.0 Hz, 1H), 6.43 (d, J = 15.9 Hz, 1H), 5.37 (s, 2H). 13C NMR (101 MHz, DMSO) δ 169.85,
162.86, 158.14, 147.48, 143.85, 137.72, 137.28, 137.24, 136.35, 130.15, 129.45, 129.03, 128.31,
127.97, 127.39, 127.34, 127.06, 126.77, 120.97, 120.42, 55.89. HRMS (AP-ESI) m/z Calcd for
C26H21N3O3 [M+H]+ 424.1656, found: 424.1661.
4.1.43
(E)-N-(4-bromophenyl)-4-(3-(hydroxyamino)-3-oxoprop-1-en-1-yl)-N-(quinolin-2-ylmethyl)be
nzamide (11b)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 33%
yield. mp：192-193 °C. 1H NMR (400 MHz, DMSO) δ 10.77 (s, 1H), 9.07 (s, 1H), 8.36 (d, J = 8.5
Hz, 1H), 7.96 (dd, J = 7.7, 4.3 Hz, 2H), 7.78 – 7.72 (m, 1H), 7.61 (d, J = 8.3 Hz, 1H), 7.59 – 7.56
(m, 1H), 7.49 (d, J = 8.3 Hz, 2H), 7.46 – 7.35 (m, 6H), 7.23 (d, J = 8.7 Hz, 2H), 6.46 (d, J = 15.8
Hz, 1H), 5.36 (s, 2H). 13C NMR (101 MHz, DMSO) δ 169.75, 162.89, 157.84, 147.48, 143.29,
137.65, 137.29, 136.95, 136.60, 132.33, 130.18, 130.01, 129.49, 129.05, 128.32, 127.48, 127.41,

ACCEPTED MANUSCRIPT
126.81, 121.26, 120.43, 119.70, 55.66. HRMS (AP-ESI) m/z Calcd for C26H20BrN3O3 [M+H]+
502.0761, found: 502.0769.
4.1.44
(E)-4-(3-(hydroxyamino)-3-oxoprop-1-en-1-yl)-N-(4-methoxyphenyl)-N-(quinolin-2-ylmethyl)ben
zamide (11c)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 32%
yield. mp：188-190 °C. 1H NMR (400 MHz, DMSO) δ 10.78 (s, 1H), 9.08 (s, 1H), 8.36 (d, J = 8.5
Hz, 1H), 7.96 (dd, J = 7.7, 5.0 Hz, 2H), 7.77 – 7.71 (m, 1H), 7.64 (d, J = 8.4 Hz, 1H), 7.61 – 7.55
(m, 1H), 7.49 – 7.34 (m, 5H), 7.18 (d, J = 8.6 Hz, 2H), 6.74 (d, J = 8.8 Hz, 2H), 6.44 (d, J = 15.8
Hz, 1H), 5.32 (s, 2H), 3.62 (s, 3H). 13C NMR (101 MHz, DMSO) δ 169.85, 162.92, 158.21,
157.99, 147.46, 137.72, 137.47, 137.22, 136.44, 136.17, 130.14, 129.38, 129.03, 128.31, 127.39,
127.31, 126.76, 121.01, 120.49, 114.59, 56.03, 55.57. HRMS (AP-ESI) m/z Calcd for C27H23N3O4
[M+H]+ 454.1761, found: 454.1755.
4.1.45
(E)-N-((8-chloroquinolin-2-yl)methyl)-4-(3-(hydroxyamino)-3-oxoprop-1-en-1-yl)-N-(4-methoxy
phenyl)benzamide (11d)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 54%
yield. mp：178-179 °C. 1H NMR (400 MHz, DMSO) δ 10.74 (s, 1H), 9.11 (s, 1H), 8.44 (d, J = 8.5
Hz, 1H), 8.02 – 7.91 (m, 2H), 7.71 (d, J = 8.3 Hz, 1H), 7.57 (t, J = 7.8 Hz, 1H), 7.50 – 7.28 (m,
7H), 6.77 (d, J = 8.9 Hz, 2H), 6.45 (d, J = 15.8 Hz, 1H), 5.34 (s, 2H), 3.64 (s, 3H). 13C NMR (101
MHz, DMSO) δ 169.75, 162.86, 159.04, 158.05, 143.34, 137.78, 137.70, 137.61, 136.93, 136.08,
132.44, 130.23, 129.66, 129.25, 128.87, 127.87, 127.29, 126.95, 121.58, 120.88, 114.51, 55.59,
49.08. HRMS (AP-ESI) m/z Calcd for C27H22ClN3O4 [M+H]+ 488.1372, found: 488.1364.
4.1.46
(E)-N-((6-bromoquinolin-2-yl)methyl)-4-(3-(hydroxyamino)-3-oxoprop-1-en-1-yl)-N-phenylbenza
mide (11e)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 39%
yield. mp：138-141 °C. 1H NMR (400 MHz, DMSO) δ 10.73 (s, 1H), 9.04 (s, 1H), 8.35 (d, J = 8.6
Hz, 1H), 8.27 (s, 1H), 7.94 – 7.82 (m, 2H), 7.69 (d, J = 8.5 Hz, 1H), 7.48 – 7.32 (m, 5H), 7.28 –
7.15 (m, 4H), 7.10 (t, J = 7.2 Hz, 1H), 6.44 (d, J = 19.9 Hz, 1H), 5.35 (s, 2H). 13C NMR (101
MHz, DMSO) δ 169.84, 162.90, 158.94, 146.08, 143.82, 137.77, 137.21, 136.50, 136.35, 133.17,
131.24, 130.32, 129.47, 128.73, 127.99, 127.33, 127.11, 121.41, 120.98, 119.59, 55.87. HRMS
(AP-ESI) m/z Calcd for C26H20BrN3O3 [M+H]+ 502.0761, found: 502.0772.
4.1.47
(E)-4-(3-(hydroxyamino)-3-oxoprop-1-en-1-yl)-N-((8-methoxyquinolin-2-yl)methyl)-N-phenylben
zamide (11f)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 29%
yield. mp：198-200 °C. 1H NMR (400 MHz, DMSO) δ 10.75 (s, 1H), 9.05 (s, 1H), 8.30 (d, J = 8.5
Hz, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.50 – 7.42 (m, 5H), 7.38 (d, J = 15.8 Hz, 1H), 7.28 (d, J = 7.7
Hz, 2H), 7.19 (dd, J = 9.4, 6.0 Hz, 3H), 7.09 (t, J = 7.4 Hz, 1H), 6.44 (d, J = 15.8 Hz, 1H), 5.35 (s,

ACCEPTED MANUSCRIPT
2H), 3.96 (s, 3H). 13C NMR (101 MHz, DMSO) δ 170.00, 162.94, 156.49, 155.38, 143.95, 139.28,
137.80, 137.54, 137.04, 136.23, 129.44, 129.38, 128.43, 128.06, 127.28, 127.01, 120.94, 120.59,
119.77, 109.23, 56.20, 55.90. HRMS (AP-ESI) m/z Calcd for C27H23N3O4 [M+H]+ 454.1689,
found: 454.1699.
4.1.48
(E)-N-hydroxy-3-(4-(2-oxo-2-(phenyl(quinolin-2-ylmethyl)amino)ethyl)phenyl)acrylamide (11g)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 37%
yield. mp：160-164 °C. 1H NMR (400 MHz, DMSO) δ 10.73 (s, 1H), 9.02 (s, 1H), 8.33 (d, J = 8.5
Hz, 1H), 7.98 – 7.88 (m, 2H), 7.74 (t, J = 7.5 Hz, 1H), 7.57 (t, J = 13.7, 5.9 Hz, 2H), 7.53 (d, J =
8.4 Hz, 1H), 7.46 (d, J = 8.3 Hz, 2H), 7.43 – 7.27 (m, 5H), 7.16 (d, J = 7.4 Hz, 2H), 6.42 (d, J =
15.8 Hz, 1H), 5.15 (s, 2H), 3.57 (s, 2H). 13C NMR (101 MHz, DMSO) δ 170.50, 163.31, 158.18,
147.40, 143.09, 138.61, 137.76, 137.12, 133.50, 130.28, 130.13, 130.00, 128.95, 128.80, 128.39,
128.30, 127.75, 127.36, 126.74, 120.53, 119.08, 55.38. HRMS (AP-ESI) m/z Calcd for
C27H23N3O3 [M+H]+ 438.1812, found: 438.1816.
4.1.49
(E)-3-(4-(2-(((6-bromoquinolin-2-yl)methyl)(phenyl)amino)-2-oxoethyl)phenyl)-N-hydroxyacryla
mide (11h)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 46%
yield. mp：170-173 °C. 1H NMR (400 MHz, DMSO) δ 8.33 (d, J = 8.6 Hz, 1H), 8.26 (s, 1H), 7.86
(d, J = 1.9 Hz, 2H), 7.59 (d, J = 8.5 Hz, 1H), 7.47 (d, J = 8.4 Hz, 2H), 7.44 – 7.29 (m, 6H), 7.17
(d, J = 7.6 Hz, 2H), 6.50 (d, J = 15.8 Hz, 1H), 5.15 (s, 2H), 3.58 (s, 2H). 13C NMR (101 MHz,
DMSO) δ 170.53, 163.20, 158.98, 146.00, 143.06, 138.32, 137.64, 136.39, 133.56, 133.16, 131.14,
130.30, 130.24, 130.01, 128.78, 128.69, 128.42, 127.71, 121.50, 119.56, 119.30, 55.36. HRMS
(AP-ESI) m/z Calcd for C27H22BrN3O3 [M+H]+ 516.0917, found: 516.0928.
4.1.50
(E)-N-hydroxy-3-(4-(2-(((6-methoxyquinolin-2-yl)methyl)(phenyl)amino)-2-oxoethyl)phenyl)acry
lamide (11i)
Using the synthetic method for 4a, the target compound was obtained as a white solid in 33%
yield. mp：190-191 °C. 1H NMR (400 MHz, DMSO) δ 10.77 (s, 1H), 9.04 (s, 1H), 8.22 (d, J = 8.5
Hz, 1H), 7.82 (d, J = 8.9 Hz, 1H), 7.54 – 7.43 (m, 3H), 7.43 – 7.26 (m, 8H), 7.16 (d, J = 7.2 Hz,
2H), 6.45 (d, J = 15.8 Hz, 1H), 5.11 (s, 2H), 3.88 (s, 3H), 3.56 (s, 2H). 13C NMR (101 MHz,
DMSO) δ 170.46, 168.12, 163.34, 157.56, 155.43, 143.31, 143.03, 138.54, 137.75, 136.03, 133.53,
130.31, 130.27, 129.97, 128.80, 128.42, 127.74, 122.43, 120.79, 119.20, 106.18, 55.95, 55.16.
HRMS (AP-ESI) m/z Calcd for C28H25N3O4 [M+H]+ 468.1918, found: 468.1926.
4.1.51 Methyl 4-(((quinolin-2-ylmethyl)amino)methyl)benzoate (13a)
Using the synthetic method for 4a, the target compound was obtained as a yellow oil in 40%
yield. 1H NMR (400 MHz, CDCl3) δ 8.11 (d, J = 8.4 Hz, 1H), 8.06 (d, J = 8.5 Hz, 1H), 8.01 (d, J
= 8.3 Hz, 2H), 7.80 (d, J = 8.1 Hz, 1H), 7.73 – 7.67 (m, 1H), 7.52 (t, J = 11.6, 4.6 Hz, 1H), 7.48
(d, J = 8.3 Hz, 2H), 7.44 (d, J = 8.4 Hz, 1H), 4.11 (s, 2H), 3.97 (s, 2H), 3.91 (s, 3H).

ACCEPTED MANUSCRIPT
4.1.52 Methyl 4-((((8-chloroquinolin-2-yl)methyl)amino)methyl)benzoate (13b)
Using the synthetic method for 4a, the target compound was obtained as a yellow oil in 37%.
1H NMR (400 MHz, CDCl3) δ 7.91 (d, J = 8.2 Hz, 3H), 7.85 – 7.80 (m, 2H), 7.65 (dd, J = 9.0, 2.2
Hz, 1H), 7.36 (t, J = 8.3 Hz, 3H), 3.99 (s, 2H), 3.87 (s, 2H), 3.81 (s, 3H).
4.1.53 Methyl 4-((((6-bromoquinolin-2-yl)methyl)amino)methyl)benzoate(13c)
Using the synthetic method for 4a, the target compound was obtained as a yellow oil in 53%
yield. 1H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 8.2 Hz, 3H), 7.94 (d, J = 2.1 Hz, 1H), 7.92 (d, J
= 9.0 Hz, 1H), 7.75 (dd, J = 9.0, 2.2 Hz, 1H), 7.46 (t, J = 8.3 Hz, 3H), 4.09 (s, 2H), 3.97 (s, 2H),
3.91 (s, 3H).
4.1.54 Methyl 4-((((6-methoxyquinolin-2-yl)methyl)amino)methyl)benzoate(13d)
Using the synthetic method for 4a, the target compound was obtained as a yellow oil in 44%
yield. 1H NMR (400 MHz, CDCl3) δ 7.99 (d, J = 8.2 Hz, 2H), 7.94 (d, 2H), 7.44 (d, J = 8.1 Hz,
2H), 7.37 – 7.31 (m, 2H), 7.01 (d, J = 2.6 Hz, 1H), 4.04 (s, 2H), 3.91 (s, 2H), 3.87 (s, 3H), 3.86 (s,
3H).
4.1.55 Methyl 4-((benzyl(quinolin-2-ylmethyl)amino)methyl)benzoate (15a)
Using the synthetic method for 5a, the target compound was obtained as a colourless oil in 33%
yield. 1H NMR (400 MHz, CDCl3) δ 8.13 (d, J = 8.5 Hz, 1H), 8.04 (d, J = 8.5 Hz, 1H), 7.99 (d, J
= 8.3 Hz, 2H), 7.79 (d, J = 8.1 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.71 – 7.65 (m, 1H), 7.54 – 7.47
(m, 3H), 7.41 (d, J = 7.1 Hz, 2H), 7.33 (t, J = 7.4 Hz, 2H), 7.23 (d, 1H), 3.90 (s, 3H), 3.89 (s, 2H),
3.69 (s, 2H), 3.64 (s, 2H).
4.1.56 N-hydroxy-4-(((quinolin-2-ylmethyl)amino)methyl)benzamide (14a)
Using the synthetic method for 6a, the target compound was obtained as a white solid in 44%
1
yield. mp：186-188 °C. H NMR (400 MHz, DMSO) δ 11.32 (s, 1H), 9.69 (s, 1H), 9.12 (s, 1H),
8.47 (d, J = 8.5 Hz, 1H), 8.09 (d, J = 8.4 Hz, 1H), 8.05 (d, J = 8.0 Hz, 1H), 7.88 – 7.79 (m, 3H),
7.68 (t, J = 7.4 Hz, 3H), 7.62 (d, J = 8.5 Hz, 1H), 4.54 (s, 2H), 4.37 (s, 2H). 13C NMR (101 MHz,
DMSO) δ 164.08, 153.51, 147.01, 137.69, 135.55, 133.62, 130.76, 130.68, 128.88, 128.57, 127.69,
127.58, 127.46, 121.19, 50.67, 50.22. HRMS (AP-ESI) m/z Calcd for C18H17N3O2 [M+H]+
308.1394, found: 308.1389.
4.1.57 4-((((8-Chloroquinolin-2-yl)methyl)amino)methyl)-N-hydroxybenzamide (14b)
Using the synthetic method for 6a, the target compound was obtained as a white solid in 33%
yield. mp：147-150 °C. 1H NMR (400 MHz, DMSO) δ 11.32 (s, 1H), 9.80 (s, 1H), 9.10 (s, 1H),
8.56 (d, J = 8.5 Hz, 1H), 8.04 (dd, J = 7.9, 3.2 Hz, 2H), 7.84 (d, J = 8.2 Hz, 2H), 7.78 (d, J = 8.5
Hz, 1H), 7.71 (d, J = 8.2 Hz, 2H), 7.66 (t, J = 7.9 Hz, 1H), 4.61 (s, 2H), 4.49 (s, 2H). 13C NMR
(101 MHz, DMSO) δ 164.07, 154.56, 142.93, 138.40, 135.58, 133.68, 132.32, 130.78, 130.73,
129.16, 128.07, 127.78, 127.61, 122.29, 50.63, 50.37. HRMS (AP-ESI) m/z Calcd for
C18H16ClN3O2 [M+H]+ 342.1004, found: 342.1006.
4.1.58 4-((((6-Bromoquinolin-2-yl)methyl)amino)methyl)-N-hydroxybenzamide (14c)
Using the synthetic method for 6a, the target compound was obtained as a white solid in 36%

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yield. mp：110-114°C. 1H NMR (400 MHz, DMSO) δ 11.15 (s, 1H), 8.99 (s, 1H), 8.32 (d, J = 8.6
Hz, 1H), 8.26 (d, J = 1.8 Hz, 1H), 7.91 (d, J = 9.0 Hz, 1H), 7.86 (dd, J = 2.0 Hz, 1H), 7.72 (dd, J
= 8.3, 4.6 Hz, 3H), 7.44 (d, J = 8.1 Hz, 2H), 3.96 (s, 2H), 3.79 (s, 2H). 13C NMR (101 MHz,
DMSO) δ 164.34, 162.41, 146.06, 144.02, 136.01, 132.90, 131.72, 131.12, 130.27, 128.75, 128.28,
127.20, 121.93, 119.22, 54.89, 52.60. HRMS (AP-ESI) m/z Calcd for C18H16BrN3O2 [M+H]+
386.0499, found: 386.0497.
4.1.59 N-hydroxy-4-((((6-methoxyquinolin-2-yl)methyl)amino)methyl)benzamide (14d)
Using the synthetic method for 6a, the target compound was obtained as a white solid in 41%
yield. mp：174-177 °C. 1H NMR (400 MHz, DMSO) δ 11.35 (s, 1H), 9.94 (s, 1H), 9.10 (s, 1H),
8.33 (d, J = 8.5 Hz, 1H), 7.97 (d, J = 9.1 Hz, 1H), 7.82 (d, J = 8.1 Hz, 2H), 7.73 – 7.57 (m, 3H),
7.72 – 7.57 (m, 3H), 7.45 (dd, J = 9.4, 7.1 Hz, 2H), 4.44 (d, J = 8.0 Hz, 2H), 4.33 (s, 2H), 3.91 (s,
3H). 13C NMR (101 MHz, DMSO) δ 164.08, 158.00, 150.65, 143.06, 136.45, 135.50, 133.60,
130.77, 130.35, 128.93, 127.57, 123.03, 121.57, 106.33, 56.07, 50.63, 50.14. HRMS (AP-ESI)
m/z Calcd for C19H19N3O3 [M+H]+ 338.1499, found: 338.1502.
4.1.60 4-((Benzyl(quinolin-2-ylmethyl)amino)methyl)-N-hydroxybenzamide (16a)
Using the synthetic method for 6a, the target compound was obtained as a white solid in 45%
yield. mp：90-92°C. 1H NMR (400 MHz, DMSO) δ 11.15 (s, 1H), 8.95 (s, 1H), 8.36 (d, J = 8.5 Hz,
1H), 8.00 – 7.92 (m, 2H), 7.76 – 7.70 (m, 4H), 7.57 (t, J = 7.5 Hz, 1H), 7.51 (d, J = 8.1 Hz, 2H),
7.43 (d, J = 7.3 Hz, 2H), 7.35 (t, J = 7.5 Hz, 2H), 7.26 (t, J = 7.3 Hz, 1H), 3.81 (s, 2H), 3.64 (s,
2H), 3.60 (s, 2H). 13C NMR (101 MHz, CDCl3) δ 166.69, 160.48, 147.09, 143.52, 138.69, 136.84,
129.94, 129.68, 129.03, 128.84, 128.42, 128.37, 127.58, 127.38, 127.20, 127.03, 126.34, 120.87,
60.08, 58.49, 57.87. HRMS (AP-ESI) m/z Calcd for C25H23N3O2 [M+H]+ 398.1863, found:
398.1856.
4.2. In vitro HDAC enzymatic assay
4.2.1. HeLa nuclear extracts
HDAC inhibitory activities of all the final products and the positive control drug Vorinostat
were evaluated by the Color de Lys™ assay (BMLAK501, Enzo®Life Sciences) including
HDAC1&2. Based on the HDAC kit instruction, HeLa cell nucleus extracts (HDAC1&2),
substrate and tested compounds (including positive control compound) were diluted to needed
concentrations. First, HeLa cell nucleus extracts (15µL/well) were incubated at 37°C for 5 min in
the 96-well plate with different concentrations of tested compounds (10µL /well). Then substrate
(25µL/well) was added and the mixture was incubating at 37°C for 30 min. At the end, the mixture
of Color de Lys Developer and TSA (50µL/well) was added. After incubation for 30 min, the
ultraviolet absorption was measured on a microtiterplate reader at 405 nm. The inhibition rates
were calculated from the ultraviolet absorption of inhibited wells and control ones. A regression
analysis method between the concentration and inhibition rate was used to give the IC50 values.
4.2.2. HDAC1, HDAC6 and HDAC8
All of the enzymatic reactions were conducted at 37°C for 30 minutes. The 50ul reaction
mixture contains 25 mM Tris, pH 8.0, 1 mM MgCl2, 0.1 mg/ml BSA, 137mM NaCl, 2.7 mM KCl,
HDAC and the enzyme substrate. The compounds were diluted in 10% DMSO and 5µL of the

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dilution was added to a 50µL reaction so that the final concentration of DMSO is 1% in all of
reactions. The assay was performed by quantitating the fluorescent product amount of in solution
following a enzyme reaction. Fluorescence is then analyzed with an excitation of 350-360 nm and
an emission wavelength of 450-460 nm at SpectraMax M5 microtiter plate reader. The IC50 values
were calculated using nonlinear regression with normalized dose-response fit in Prism GraphPad
software.
4.3. MTT assay
All cell lines were cultured in RPMI1640 medium (10% FBS) at 37°C in a 5% CO2 humidified
incubator. Briefly, Cancer cells were plated at 4000–5000 cells per well (100 µL/well) in 96-well
plates for 8 h and then treated with different concentrations of compounds (100 µL/well) for 48 h.
0.5% MTT solution (10 µL/well) was added and incubation for 4 h. DMSO (150 µL/well) was
added to extract formazan formed from MTT. Absorbance was determined using a microtiter-plate
reader at 570 nm. The IC50 values were calculated according to the inhibition ratios.
4.4. Whole-cell HDAC inhibition assay
The cellular HDAC assay was based on such assay described on patent WO20080952A1 with
minor modifications. Briefly, human chronic myelogenous leukemia cell K562 were seeded into a
96-well tissue culture plates (Corning, Germany) at a density of 1.2×104 cells/well and incubation
for 24 h. Then cells were treated with various concentrations of test compounds for 3 h. Substrates
were added which Boc-Lys (acetyl)-AMC was used to measuring class I HDAC activity or
Boc-Lys (triflouroacetyl)-AMC for measuring HDAC class IIa activity (final concentration: 0.2
mM). 3 h later under cell culture conditions, stop solution (50 mM Tris–HCl (pH 8.0), 137 mM
NaCl, 2.7 mM KCl, 1 mM MgCl2, 1% NP-40, 2.0 mg/mL trypsin, 10µM TSA) was added and the
mixture was incubated for another 3 h. Fluorescence intensity was measured using a microplate
reader at excitation and emission wavelengths of 360 and 470 nm, respectively.
4.5. Computational details
The crystal structure of HDAC1 (PDB: 5ICN [29]) was used in current study. PDB2PQR server
was used to determine the protonation states of charged residues in HDAC1[30]. Specially, the
protonation state of catalytic residues are determined based on our previous QM/MM study[31].
The initial structure of compound 4a was built and minimized using Sybyl-X package. Molecular
docking was performed using Autodock Vina[28] and the top-scored result was used as the
starting conformation for MD simulation.
MD simulation was performed using Amber14 package[32] with Amber99SB[33] and TIP3P
water model. Partial charges for compound 4a was fit with HF/6-31G(d) calculations using
Gaussian 09 package[34]. The HDAC1-4a system was neutralized with Cl- counterions and
solvated with explicit TIP3P water. Short-long effective function2 (SLEF2[35]) was introduced to
describe charge interactions between the zinc ion and all other atoms. After a series of energy
minimization and equilibration, 20ns standard molecular dynamics simulation was performed with
periodic boundary condition. Other parameters were default values. All saved MD snapshots were
analyzed using AmberTools 15. Figures are created using PyMOL (The PyMOL Molecular
Graphics System, Version 1.7. 4 Schrödinger, LLC.).

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Acknowledgments
This research was supported by the National Natural Science Foundation China (No. 81373281).
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1. Novel quinoline-based N-hydroxycinnamamides were designed and synthesized.
2. Compound 4a showed better inhibitory activity for class I HDAC than Vorinostat.
3. Compound 4e exhibited excellent anti-proliferative activity against K562.
4. Compound 4a and 4e could promote cell apoptosis in vitro.