Synthesis and biological evaluation of β2-adrenoceptor agonists bearing the 2-amino-2-phenylethanol scaffold

Article abstract of DOI:10.1016/j.ejmech.2018.04.041

A new series of β₂-adrenoceptor agonists bearing the 2-amino-2-phenylethanol scaffold was synthesized. Evaluation of the compounds using cell assays and an in vitro guinea pig trachea relaxation assay showed that 8-hydroxy-5-(2-hydroxy-1-((4-hy

Full text of DOI:10.1016/j.ejmech.2018.04.041

Accepted Manuscript
Synthesis and biological evaluation of β -adrenoceptor agonists bearing the 2-
2
amino-2-phenylethanol scaffold
Xinyue Ge, Anthony Yiu-Ho Woo, Gang Xing, Yali Lu, Yongmei Mo, Ying Zhao, Yi
Lan, Jinyan Li, Haining Yan, Li Pan, Yuyang Zhang, Bin Lin, Maosheng Cheng
PII:
S0223-5234(18)30375-1
10.1016/j.ejmech.2018.04.041
EJMECH 10390
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 22 January 2018
Revised Date: 6 April 2018
Accepted Date: 19 April 2018
Please cite this article as: X. Ge, A.Y.-H. Woo, G. Xing, Y. Lu, Y. Mo, Y. Zhao, Y. Lan, J. Li, H. Yan,
L. Pan, Y. Zhang, B. Lin, M. Cheng, Synthesis and biological evaluation of β -adrenoceptor agonists
2
bearing the 2-amino-2-phenylethanol scaffold, European Journal of Medicinal Chemistry (2018), doi:
10.1016/j.ejmech.2018.04.041.
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ACCEPTED MANUSCRIPT

ACCEPTED MANUSCRIPT
Synthesis and biological evaluation of β -adrenoceptor agonists bearing the
2
2
-amino-2-phenylethanol scaffold
a,b,1
c,1
a,b
c
a,b
Xinyue Ge , Anthony Yiu-Ho Woo , Gang Xing , Yali Lu , Yongmei Mo , Ying
a,b
c
a,b
a,b
a,b
c
a,b**
Zhao , Yi Lan , Jinyan Li , Haining Yan , Li Pan , Yuyang Zhang , Bin Lin
,
a,b*
Maosheng Cheng
a Department of Medicinal Chemistry, School of Pharmaceutical Engineering,
Shenyang Pharmaceutical University, Shenyang 110016, China.
b Key Laboratory of Structure-Based Drug Design and Discovery of Ministry
of Education, Shenyang Pharmaceutical University, Shenyang 110016, China.
c Department of Pharmacology, School of Life Sciences and Biopharmaceutics,
Shenyang Pharmaceutical University, Shenyang 110016, China.
*
Corresponding author.
**
Corresponding author.
Email address: mscheng@syphu.edu.cn (Maosheng Cheng)，randybinlin@gmail.com
Bin Lin);
Abstract
(
A new series of β -adrenoceptor agonists bearing the 2-amino-2-phenylethanol
2
scaffold was synthesized. Evaluation of the compounds using cell assays and an in
vitro
-hydroxy-5-(2-hydroxy-1-((4-hydroxyphenethyl)amino)ethyl)quinolin-2(1H)-one
compound 5j) has the best pharmacological profile among all the evaluated
guinea
pig
trachea
relaxation
assay
showed
that
8
(
compounds. The (S)-isomer of 5j was subsequently found to be the active
enantiomer with a promising EC50 value of 1.26 nM in stimulating
β2-adrenoceptor-mediated cAMP accumulation and a substantially higher selectivity
for the β than for the β subtype. The putative binding mode of (S)-5j revealed by
2
1
molecular docking of the β -adrenoceptor resembles that in agonist binding. Taken
2

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together, these results showed that compound (S)-5j is a promising hit worthy of
further study for the development of β -adrenoceptor agonists.
2
Key words: β -Adrenoceptor agonists, Bronchodilators, Trantinterol, Indacaterol
2
1
. Introduction
Asthma and chronic obstructive pulmonary disease (COPD) are the most
common chronic respiratory diseases that affect millions of people worldwide [1]. Up
to 300 million people suffer from asthma globally, and the prevalence of asthma has
been increasing by 50% every decade [2]. COPD is a leading cause of morbidity and
mortality with a prevalence estimated currently at 9–10% in adults aged 40 years or
older [3]. The World Health Organization (WHO) estimated back in 2006 that by
2
020 COPD would be the third leading cause of mortality and the fifth leading cause
of morbidity in the world [4]. Both of these diseases can be treated with
bronchodilators, such as β -adrenoceptor agonists and muscarinic antagonists.
2
Monotherapy or combination therapy comprising these bronchodilators and inhaled
corticosteroids are known to produce relief or reduce the frequency of exacerbation.
The more widespread use of these pharmaceutical agents has been one of the major
means to lower the burden of chronic respiratory diseases [5].
The β -adrenorcecptor, a prototypical class A G protein-coupled receptor, is
2
widely distributed in the respiratory tract, especially in airway smooth muscles.
Agonists of this receptor exert a bronchodilatory effect through relaxation of airway
smooth muscles. Activation of the β -adrenorcecptor induces an increase of
2
2
+
cytoplasmic cAMP concentration and subsequently a decrease of cytoplasmic Ca
concentration, which results in a lower muscle tone [6, 7]. Bronchodilator β -agonists
2
may be broadly classified into three generations based on their onset of action and

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duration of action. The first generation or short-acting β -agonists with no more than
2
4
-6 h duration of action, such as salbutamol and terbutaline, are used mainly for quick
asthma relief [8, 9]. The second generation agents are long-acting with a duration of
action of about 12 h. Salmeterol, formoterol and other agents in this class are used in
persistent asthma and COPD [8]. The ultralong-acting β -agonists most recently
2
developed such as Novartis’ indacaterol [10], Boehringer-Ingelheim’s olodaterol [11]
and GlaxoSmithKline’s vilanterol [12] have duration of action that last for a whole
day.
In recent years, at least six candidates of β -adrenoceptor agonists are under
2
clinical development. These include Pfizer’s PF-610355 [1], Theravance’s TD-5471
and TD-4306 [13, 14], Almirall’s abediterol [15], Theravance’s milveterol [16] and
trantinterol (Fig. 1) developed by our group. Trantinterol is different from the other
candidates because it bears a 2-amino-2-phenylethanol scaffold rather than a
2
-amino-1-phenylethanol scaffold found in classical β -agonists. Pharmacological
2
characterization of trantinterol has demonstrated that it is a selective β -agonist with
2
strong biological activities [17].
Fig. 1 Structures of recent β -adrenoceptor agonists
2
In the present study, the head group (8-hydroxy-1H-quinolin-2-one) and the tail
group (aminoaryl portion) of indacaterol were combined with the 2-aminoethan-1-ol

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core of trantinterol to yield the lead compound 5a (Fig. 1) which had an EC value of
5
0
3
57.0 nM in an assay for β -adrenoceptor-mediated cAMP accumulation. To optimize
2
biological activity, a series of 5a analogs with different tail groups were subsequently
synthesized and biologically evaluated.
2. Results and discussion
2.1. Chemistry
The syntheses of the target compounds 5a-x are illustrated in Scheme 1. The
key intermediate 1 was synthesized according to a general procedure [18]. Reduction
of 1 with sodium borohydride afforded 2, which was subjected to intramolecular
nucleophilic substitution to afford the epoxide 3 in the presence of K CO .
2
3
Intermediates 4a-x were prepared by refluxing 3 with different primary amines in the
presence of zinc chloride as catalyst and acetonitrile as solvent. Target compounds
5
a-x were synthesized from 4a-x by hydrogenation with Pd/C in methanol.
Scheme 1. Synthetic route of the title compounds 5a-5x. Reagents and conditions: (a) NaBH , MeOH, rt., 1h,
4
9
4.5%; (b) K CO , acetone, reflux, 6h, 98.2%; (c) RNH , ZnCl , CH CN, reflux, 12h, 16.5-30.2%; (d) Pd/C, H ,
2 3 2 2 3 2
MeOH, rt., 2h, 93.2-99.8%.
2
.2. Biological tests

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2
.2.1 Agonistic effects of the compounds on various β-adrenoceptor subtypes
To evaluate the activities of the compounds at the β , β and β -adrenoceptors,
2
3
1
compounds in Table 1 were tested for their abilities to cause cAMP accumulation in
human embryonic kidney 293 (HEK293) cells expressing human β , β or
2
3
β1-adrenoceptors.
Our lead compound 5a was found to be a modest β -adrenoceptor agonist with a
2
pEC value of 6.45 in the cell assay targeting the β -adrenoceptor. Modifications on
50
2
the tail group of 5a led to a series of compounds. Initially, alkylbenzene adducts with
different chain lengths were examined. The compound with an ethylbenzene (5b)
exhibited a higher potency than the one with an N-propylbenzene (5c) or an
N-butylbenzene (5d), with pEC values of 8.8, 8.0 and 7.9, respectively. Replacement
5
0
of the phenyl group in 5b with an indo-3-yl group resulted in compound 5e with a
lower activity. Analogs of 5b containing an alkoxyphenyl tail similar to the one in
salmeterol (5f and 5g) had no improvement in activity compared to 5b. Compounds
5
b and 5c only weakly stimulated β -adrenoceptors. Their selectivity for the β - over
1 2
β1-adrenoceptor were 520 and 80, respectively, in terms of fold change in the
respective EC50 values of the assays for the two targets. In contrast, the selectivity of
the non-selective β-agonist isoprenaline was 10. Based on the data above, compound
5
b was selected for further modification by aliphatic or aromatic substitutions.
Table1.
Activities of the compounds in stimulating cAMP accumulation in HEK293 cells
expressing the β , β or β -adrenoceptors. Isoprenaline was used as a reference.
2
3
1
a
a
a
b
Compound
R
β pEC
β pEC
β pEC
β2-β1
2
50
1
50
3
50

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5
5
5
5
5
5
5
a
b
c
d
e
f
6.5±0.6
8.8±0.4
8.0±0.2
7.9±0.2
6.5±0.5
6.6±0.3
8.3±0.2
9.5±0.1
6.08±0.3
6.08±0.5
<5
<5
<5
2.72
1.89
>2.86
g
<5
>3.31
0.99
isoprenaline
8.5±0.1
7.27
2 1 3
a Stimulation of cAMP accumulation in HEK293 cells expressing human β , β or β -adrenoceptors, pEC50 is the negative logarithm of
the molar drug concentration that produces a cAMP response equal to 50% of its maximal response.
2 1 2 1
b Selectivity for β over β , expressed as pEC50 at the β -adrenoceptor - pEC50 at the β -adrenoceptor.
Derivatives of 5b with different substituents such as hydrogen bond acceptors,
hydrogen bond donors and halogen atoms on the ethylbenzene tail were synthesized
(
Table2). Substitution on the benzene ring with a hydroxy group at different positions
5h, 5i and 5j), a meta-methoxy group (5k) or a para-methoxy group (5m) decreased
(
β2-agonistic activity, whereas substitution with an ortho-methoxy group resulted in a
compound (5l) with a similar β -agonistic activity as the parent compound.
2
Substitution with two methyl groups or a phenyl group on the ethyl moiety yielded
two compounds 5q and 5r with lower activities. For the activities at the
β1-adrenoceptor, the pEC50 values of most of these compounds were less than 5.
Comparing the activities of the compounds at different β-adrenoceptor subtypes,
compounds 5j and 5k were identified to be more potent (pEC >8) and more selective
5
0
to the β -adrenoceptor (fold change in the respective EC values for β over β >2000)
2
50
2
1
among the compounds studied so far. Although some chlorine-containing derivatives

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of 5b had high activities at the β -adrenoceptor, they were discontinued in our
2
subsequent study due to low solubility in water.
Table 2
Activities of the compounds in stimulating cAMP accumulation in HEK293 cells
expressing β , β or β -adrenoceptors. Isoprenaline was used as a reference.
2
3
1
a
a
a
b
Compound R1 R2 R3 R4
β pEC
β pEC
β pEC
β2-β1
2
50
1
50
3
50
5
5
5
5
5
5
5
5
5
5
5
h
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
2-OH
3-OH
4-OH
8.0±0.3
6.1±0.3
8.5±0.1
<5
>3.03
i
j
<5
<5
<5
>3.5
>3.3
2.62
>2.71
>3.66
3.05
1.6
k
l
2-OMe 8.3±0.3
3-OMe 8.7±0.2
4-OMe 7.7±0.1
<5
6.2±0.6
<5
m
n
o
p
q
r
2-Cl
3-Cl
4-Cl
-
8.7±0.5
8.6±0.4
7.4±0.5
7.3±0.2
6.2±0.4
9.5±0.1
<5
5.5±0.5
5.8±0.4
<5
Me Me H
Ph -
>2.28
H
H
isoprenaline
8.5±0.1
0.99±0.1
2 1 3
a Stimulation of cAMP accumulation in HEK293 cells expressing human β , β or β -adrenoceptors, pEC50 is the negative logarithm of

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the molar drug concentration that produces a cAMP response equal to 50% of its maximal response.
2 1 2 1
b Selectivity for β over β , expressed as pEC50 at the β -adrenoceptor - pEC50 at the β -adrenoceptor.
Replacement of the 4-hydroxy substituent in the ethylbenzene tail of 5j with an
amino group yielded compound 5s with a lower potency. Based on compound 5s, a
series of amide derivatives were synthesized. As shown in Table 3, amides with a
phenyl group like compounds 5u and 5t showed higher β -agonistic activities,
2
whereas amides with an alkyl chain such as 5v, 5w and 5x had similar or lower
activities as compared to 5s. It is possible that the additional phenyl group could form
extra π-π interactions with amino acid residues farther away from the orthosteric
binding site. Nevertheless, compound 5u, with a potency similar to those of 5j and 5k,
were not included in subsequent characterization because its yield was too low.
Table 3
Activities of the compounds in stimulating cAMP accumulation in HEK293 cells
expressing β -adrenoceptors. Isoprenaline was used as a reference.
2
a
Compound
R
H
β pEC
2
50
5
5
s
t
7.1±0.1
7.3±0.2
5
u
v
8.4±0.3
7.3±0.2
5

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5
5
w
x
6.5±0.2
6.2±0.4
isoprenaline
9.5±0.1
a Stimulation of cAMP accumulation in HEK293 cells expressing human β
2
-adrenoceptors, pEC50 is the negative logarithm of the molar
drug concentration that produces a cAMP response equal to 50% of its maximal response.
2
.2.2 Study of intrinsic activity
The efficacies (i.e. the ability to cause maximal cAMP accumulation in HEK293
cells endogenously expressing human β -adrenoceptors) of selected compounds with
2
high potency and high β -selectivity were determined and expressed in terms of
2
intrinsic activities (IA), defined as the maximal responses of the test compounds
relative to the maximal response of the full β-agonist isoprenaline. By definition, the
intrinsic activity of isoprenaline is 1. As shown in Table 4, the intrinsic activities of
all these compounds were smaller than that of isoprenaline. Therefore, these
compounds are partial agonists of the human β -adrenoceptor in this
2
non-overexpressed system.
Table 4
Intrinsic activities of the compounds in HEK293 cells endogenously expressing
human β -adrenoceptors
2
Compound
Intrinsic activity
vs. isoprenaline %)
57.4±9.1
(
5
b
5
c
55.8±2.3
5
h
54.2±8.3

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5
j
46.3±7.8
48.6±4.6
52.5±7.3
5
k
5
l
isoprenaline
100.0±12.4
Intrinsic activity (IA) is expressed as a percentage of the maximal effect of isoprenaline (=100%).
2
.2.3 In vitro study of isolated guinea pig trachea relaxation
Three highly potent compounds in the cell assay, compounds 5b, 5j and 5k, were
further characterized in an in vitro smooth muscle relaxation assay using isolated
guinea pig trachea tissues. The effectiveness of the compounds in producing smooth
muscle relaxation in the assay can reflect the efficacy of the compounds as
bronchodilator drugs. Spasm induced by histamine and relaxation produced by the test
compounds were quantified by measuring muscle tones. The relaxation percentages
associated with each accumulated doses of the test compounds were calculated and
the data are shown in Fig. 2. Pharmacodynamic parameters of the test compounds are
summarized in Table 5. Emax represents the maximal effect obtained using the
saturating concentration of a test compound. Potency (pD ) represents the negative
2
logarithm of the concentration of the test compound, at which the relaxant effect is
equal to 50% of the compound’s own maximal effect. The Emax values of compounds
5
j and 5b did not differ from that of isoprenaline. However, their pD values were
2
smaller than that of isoprenaline. The Emax value of compound 5k was smaller than
that of isoprenaline, and so as its pD value. From the intrinsic activities of the test
2
compounds, it could be deduced that compounds 5b and 5j are full agonists in the
assay, whereas 5k is a partial agonist. Based on the data so far, compound 5j with
superior pharmacodynamic properties was selected for further studies.

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Fig. 2 Concentration-responses for compounds 5b, 5j, 5k and isoprenaline in relaxation of
isolated guinea pig trachea slices (n=4).
Table 5
Emax, pD and IA (vs. isoprenaline) values of compounds 5b, 5j, 5k and isoprenaline.
2
pD2
Emax (%)
IA (vs. isoprenaline)
isoprenaline
8.16±0.08
7.81±0.48
7.98±0.31
6.03±0.21*
254.48±91.49
255.34±60.30
249.93±41.83
111.13±65.60*
1.00
1.00
0.98
0.44
5
5
5
b
j
k
The pharmacodynamic parameters were determined in isolated guinea-pig trachea slices pre-contracted with
-
5
histamine (5×10 M). Data are expressed as means ± standard error of the mean (S.E.M.).* Differences between
the values (vs. those of isoprenaline) were compared by unpaired Student’s t test and P<0.05 was considered
significant
.
2
.2.4 In vitro pharmacokinetic studies
To identify the active configuration, the (R) and (S)-isomers of 5j were chirally

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separated by column chromatography. In the cell assay, the EC of (S)-5j was found
5
0
to be 1.26 nM whereas that of (R)-5j was 328 nM. The data suggest that the
S)-isomer is the active enantiomer. Compound (S)-5j was further subjected to in vitro
(
pharmacokinetic studies. It was found to be a slow substrate of drug metabolism in
human liver microsomes, with a modest intrinsic hepatic clearance (Clint = 11.5
mL/min/kg, T =108.6 min) as compared to the reference compound isoprenaline
1
/2
(
Cl = 112.3 mL/min/kg, T =12.3 min). The permeability of compound (S)-5j, in
int 1/2
terms of apical to basal (A-B, or B-A in the reverse direction) flux rate through a
monolayer of Caco-2 cells, was determined. The resultant bi-directional apparent
-6
-6
permeability coefficients (P A-B=0.43 ×10 cm/s, P B-A=0.74 ×10 cm/s)
app
app
suggest that (S)-5j possesses intrinsically poor cell permeability and that (S)-5j may
have a low oral bioavailability due to poor absorption through the gut wall. Thus,
(S)-5j appeared to exhibit an ideal pharmacokinetic profile for an inhaled agent
because the swallowed fraction of the dose of the inhaled drug may be less prone to
absorption [19].
2
2
.3 Molecular modeling
.3.1 Predicted binding mode of compound (S)-5j.
Understanding the binding mode of a lead compound in a molecular target
provides an opportunity to rationally design better drugs for the target. To explore
ligand-target interactions, compound (S)-5j was docked to the molecular model of the
crystallographic structure of the β -adrenoceptor co-crystallized with the high affinity
2
agonist hydroxybenzyl isoproterenol (HBI) (PDB: 4LDL) using the docking program

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Glide in Schrödinger Maestro. The most reasonable pose was used for the analysis of
protein-ligand interactions. For the HBI-bound β -adrenoceptor (Fig. 3A), the two
2
hydroxy groups on the catechol head of HBI formed hydrogen bonds with Asn293,
Ser207 and Ser203, which are thought to be important for agonist binding and
receptor activation. The β-OH group and the charged secondary amine moiety of HBI
formed a hydrogen bond network with Asp113 and Asn312. Furthermore, π−π
interactions and π−σ interactions were observed between HBI and residues Val114,
Val117, Phe193, Phe290 and Ile309 [20]. For the simulated binding pose of
compound (S)-5j in the binding pocket of the β -adrenoceptor (Fig. 3B), the key
2
interacting residues are the same: the aromatic head of (S)-5j was observed to interact
with Phe290 via π−π interactions, and with residues Val114 and Val117 via π−σ
interactions. The benzene moiety of the tail group was found to interact with Trp109
via π−π interaction and with Phe193 via π−σ interaction. The -OH and -NH groups on
the aromatic head formed hydrogen bonds with Ser203 and Ser207, respectively. The
β-OH group formed hydrogen bonds with Tyr316 and Asn312 and the charged
secondary amine moiety formed a hydrogen bond with Asp113 only. Besides, the
4
-OH group on the benzene tail of (S)-5j formed a hydrogen bond with Cys191
whereas the respective group in HBI had it formed with Asp192. The molecular
modeling results support experimental findings that these 2-amino-2-phenylethanol
derivatives could act as β -adrenoceptor agonists.
2

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Fig. 3 Binding modes of HBI (A) and compound (S)-5j (B).
3
. Conclusion
A series of novel β -adrenoceptor agonists based on the previously identified
2
2
-amino-2-phenylethanol scaffold have been synthesized and evaluated for agonistic
activities in β -adrenoceptor-expressing cells. Highly active compounds were further
2
characterized for receptor subtype selectivity, intrinsic activity and airway smooth
muscle relaxant effect. A hit compound (S)-5j was found to have pharmacokinetic
properties of an inhaled agent and binding features resembling a β -agonist. Taken
2
together, these preliminary data show that compound (S)-5j is a promising compound
worthy of further study for the development of β -adrenoceptor agonists.
2
4
. Experimental section
4
.1 Chemistry
Unless otherwise noted, all solvents and reagents were obtained from commercial
suppliers and were used as received. High resolution ESI-MS was performed on an
1
13
AB Sciex TripleTOF® 4600 LC/MS/MS system. H-NMR and C-NMR spectra
were recorded for compounds dissolved in DMSO-d on a Brucker AV400 NMR
6

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spectrometer. Chemical shifts (δ) are expressed in parts per million (ppm) relative to
1
the internal control (TMS). H-NMR spectra are reported in the following order:
multiplicity, approximate coupling constant (J value) in hertz (Hz), and number of
protons; signals are characterized as s (singlet), d (doublet), dd (doublet of doublets), t
(triplet), q (quartet), m (multiplet) and br (broad signal). All reactions were monitored
by TLC, using TLC plates (Silica gel 60 GF254, Merck) and UV light visualization.
Where appropriate, crude products were purified by column chromatography using
silica gel (200-300 mesh) purchased from Qingdao Haiyang Chemical Co. Ltd.
(China). All biologically evaluated compounds were >95% pure.
4
.1.1 8-(Benzyloxy)-5-(2-bromo-1-hydroxyethyl)quinolin-2(1H)-one (2)
To a solution of intermediate 1 (40 g, 0.108 mol) in methanol (800 mL) and
dichloromethane (400 mL) at 0°C, sodium borohydride was added portion-wise. After
0 min of stirring, the reaction was acidified with 2 mol/L aqueous hydrochloric acid
3
solution to pH <7. The solvent was evaporated and water (500 mL) was added,
followed by stirring for 2 h. The residue was filtered and dried to give a yellow solid
1
(
38.0 g, 94.5%). H-NMR (400 MHz, DMSO-d ) δ: 10.71 (s, 1H), 8.19 (d, J = 9.96
6
Hz, 1H), 7.38 (m, 2H), 7.31 (m, 1H), 7.19 (m, 2H), 6.56 (d, J = 9.92 Hz, 1H), 5.95 (d,
J = 4.60 Hz, 1H), 5.31 (m, 1H), 5.21 (m, 2H), 3.68 (m, 1H), 3.61 (m, 1H). MS (ESI)
+
+
m/z 374.0 [M+H] , 396.0 [M+Na] .
4
.1.2 8-(Benzyloxy)-5-(2-oxiranyl)quinolin-2(1H)-one (3)

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To a solution of intermediate 2 (38.0 g, 0.102 mol) in methanol (760 mL),
K CO (28.2 g, 0.204 mol) was added and the mixture was refluxed for 6 h. After the
2
3
reaction was completed (TLC analysis), the solvent was evaporated to give a yellow
oily liquid. Water was added and the mixture was extracted with ethyl acetate. The
organic phase was washed with water, and then with brine. The organics were dried
over sodium sulfate, filtered and concentrated to give a yellow solid (29.5 g, 99.1%).
1
H-NMR (400 MHz, DMSO-d ) δ: 10.85 (s, 1H) , 8.20 (d, J=9.84 Hz, 1H), 7.57 (d, J
6
=
7.24 Hz, 1H), 7.37 (m, 2H), 7.31 (m, 1H), 7.18 (d, J = 8.36 Hz, 1H), 6.93 (d, J =
8
2
.28 Hz, 1H), 6.61 (d, J = 9.60 Hz, 1H), 5.30 (s, 2H), 4.36 (m, 1H), 3.13 (m, 1H),
+
+
.78 (m, 2H). MS (ESI) m/z 294.1 [M+H] , 316.0 [M+Na] .
4
.1.3 General method for the synthesis of 4a-x
To a stirred solution of intermediate 3 (1.0 equiv.) and zinc chloride (0.25 equiv.)
in 150 mL acetonitrile, different primary amines (3.0 equiv.) were added to prepare
different target compounds. The reaction mixture for each was heated under reflux for
1
2 h. Then, the solvent was evaporated. The product was purified by column
chromatography (CH Cl : CH OH: NH OH = 400:10:1). After evaporation, the solid
2
2
3
4
was dissolved in 50 mL acetone, followed by acidification with a saturated solution of
hydrogen chloride in isopropanol to pH 3. Filtration of the resulting mixture gave
4
a-x as a white solid.
4
.1.3.1 8-Benzyloxy-5-(2-hydroxy-1-(isopropylamino)ethyl)quinolin-2(1H)-one (4a).
1
White solid; mp: 274.4-280.3°C; yield 12.7%; H-NMR (400 MHz, DMSO-d ) δ:
6

ACCEPTED MANUSCRIPT
1
0.90 (s, 1H), 9.80 (br, 1H), 9.60 (br, 1H), 8.31 (d, J=10.12 Hz, 1H), 7.59 (d, J = 7.20
Hz, 2H), 7.54 (m, J = 8.52 Hz, 1H), 7.29-7.40 (m, 4H), 7.95 (d, J = 7.12 Hz, 2H),
6
.66 (d, J = 9.92 Hz, 2H), 5.58 (br, 1H), 5.34 (s, 2H), 4.99 (s, 1H), 3.84 (m, 3H),
.10-3.16 (m, 3H), 2.97 (m, 1H), 2.53 (m, 4H), 1.10 (td, J = 7.48 Hz, J = 2.76 Hz,
3
1
3
1
H). C-NMR (150 MHz, DMSO-d ) δ: 160.16, 143.76, 139.38, 139.32, 136.32,
6
1
36.02, 135.88, 135.15, 129.02, 127.76, 127.34, 127.26, 123.45, 123.41, 122.57,
22.29, 120.63, 117.50, 111.57, 69.15, 61.93, 55.67, 55.37, 34.92, 34.64, 24.20,
1
+
2
4.17, 14.94, 14.90. HRMS (ESI): Calcd. for C H N NaO [M+Na] 505.2467;
31
34
2
3
found 505.2452.
4
.1.3.2 8-Benzyloxy-5-(2-hydroxy-1-(phenethylamino)ethyl)quinolin-2(1H)-one (4b).
1
White solid, mp: 276.1-277.2°C; yield 16.7%; H-NMR (400 MHz, DMSO-d ) δ:
6
1
0.90 (s, 1H), 9.86 (s, 1H), 9.53 (s, 1H), 8.23 (d, J = 10.1 Hz, 1H), 7.73 – 7.51 (m,
H), 7.38 (t, J = 7.4 Hz, 2H), 7.35 – 7.24 (m, 4H), 7.21 (t, J = 7.2 Hz, 1H), 7.14 (d, J
3
=
7.1 Hz, 2H), 6.61 (d, J = 9.9 Hz, 1H), 5.61 (s, 1H), 5.34 (s, 2H), 4.96 (s, 1H), 3.92
(d, J = 10.3 Hz, 1H), 3.81 (d, J = 8.3 Hz, 1H), 3.20 (dt, J = 15.9, 8.1 Hz, 1H), 3.13 –
1
3
2
1
1
.90 (m, 3H). C-NMR (150 MHz, DMSO-d ) δ: 161.24, 144.77, 137.73, 136.94,
6
36.49, 130.06, 129.10, 128.95, 128.83, 128.43, 128.36, 127.20, 123.68, 123.39,
21.40, 118.51, 112.68, 70.21, 62.64, 57.78, 47.17, 32.03. HRMS (ESI): Calcd. For
+
C H N O [M+H] 415.2022; found 415.2028.
26
27
2
3

ACCEPTED MANUSCRIPT
4
8
.1.3.3
-Benzyloxy-5-(2-hydroxy-1-((3-phenylpropyl)amino)ethyl)quinolin-2(1H)-one (4c).
1
White solid; mp: 238.7-239.7°C; yield 19.5%; H-NMR (400 MHz, DMSO-d ) δ:
6
1
0.92 (s, 1H), 9.54 (s, 1H), 9.23 (s, 1H), 8.23 (d, J = 10.1 Hz, 1H), 7.61 (d, J = 7.3
Hz, 2H), 7.50 (d, J = 8.4 Hz, 1H), 7.40 (t, J = 7.3 Hz, 2H), 7.32 (dd, J = 12.3, 7.9 Hz,
2
1
2
H), 7.22 (t, J = 7.2 Hz, 2H), 7.18 – 7.06 (m, 3H), 6.62 (d, J = 9.9 Hz, 1H), 5.55 (s,
H), 5.34 (s, 2H), 4.91 (s, 1H), 3.85 (dt, J = 11.1, 5.7 Hz, 1H), 3.81 – 3.70 (m, 1H),
1
3
.89 (s, 1H), 2.74 (s, 1H), 2.67 – 2.53 (m, 2H), 2.04 – 1.84 (m, 2H). C-NMR (150
MHz, DMSO-d ) δ: 161.23, 144.79, 141.03, 136.97, 130.07, 128.85, 128.77, 128.61,
6
1
3
4
4
28.44, 128.34, 126.45, 123.46, 121.31, 118.58, 112.25, 70.21, 62.61, 57.57, 45.41,
+
2.33, 27.39. HRMS (ESI): Calcd. for C H N O [M+H] 429.2178; found
2
7
29
2
3
29.2205.
.1.3.4 8-Benzyloxy-5-(2-hydroxy-1-((4-phenylbutyl)amino)ethyl)quinolin-2(1H)-one
4d).
White solid; mp: 246.2-246.9°C; yield 26.3%; H-NMR (400 MHz, DMSO-d ) δ:
(
1
6
1
0.92 (s, 1H), 9.49 (s, 1H), 9.19 (s, 1H), 8.21 (d, J = 9.9 Hz, 1H), 7.60 (d, J = 7.2 Hz,
H), 7.50 (d, J = 8.4 Hz, 1H), 7.39 (t, J = 7.1 Hz, 2H), 7.34 – 7.27 (m, 2H), 7.24 (t, J
2
=
7.1 Hz, 2H), 7.14 (t, J = 9.6 Hz, 3H), 6.62 (d, J = 9.8 Hz, 1H), 5.54 (s, 1H), 5.35 (s,
2
1
1
1
H), 4.88 (s, 1H), 3.84 (s, 1H), 3.79 (s, 1H), 2.93 (s, 1H), 2.72 (s, 1H), 2.44 (s, 2H),
1
3
.63 (s, 2H), 1.56 – 1.39 (m, 2H). C-NMR (150 MHz, DMSO-d ) δ: 161.24, 144.75,
6
41.95, 136.96, 136.40, 130.06, 128.84, 128.73, 128.68, 128.43, 128.35, 126.22,
23.60, 123.43, 121.28, 118.60, 70.20, 62.66, 57.63, 45.83, 34.92, 28.23, 25.39.

ACCEPTED MANUSCRIPT
+
HRMS (ESI): Calcd. for C H N O [M+H] 443.2335; found 443.2365.
28
31
2
3
4
8
.1.3.5
-Benzyloxy-5-(2-hydroxy-1-((2-(1H-indol-3-yl)ethyl)amino)ethyl)quinolin-2(1H)-on
e (4e).
White solid; mp: 156.8-158.3°C yield 27.6%; H-NMR (600 MHz, DMSO-d ) δ:
1
6
10.94 (s, 1H), 10.87 (s, 1H), 9.78 (s, 1H), 9.46 (s, 1H), 8.22 (d, J = 10.1 Hz, 1H), 7.64
– 7.50 (m, 3H), 7.42 – 7.36 (m, 3H), 7.35 – 7.28 (m, 3H), 7.14 (d, J = 2.3 Hz, 1H),
7.06 (t, J = 8.0 Hz, 1H), 6.94 (t, J = 7.5 Hz, 1H), 6.60 (d, J = 9.9 Hz, 1H), 5.60 (s,
1H), 5.35 (s, 2H), 5.08 – 4.90 (m, 1H), 3.99 – 3.87 (m, 1H), 3.82 (d, J = 11.3 Hz, 1H),
3.19 (d, J = 11.0 Hz, 1H), 3.11 (dd, J = 24.9, 22.9, 14.7 Hz, 2H), 3.02 (s, 1H).
1
3
C-NMR (150 MHz, DMSO-d ) δ: 161.23, 144.78, 136.95, 136.66, 136.42, 130.09,
6
1
1
28.84, 128.43, 128.35, 127.08, 123.65, 123.44, 121.63, 121.35, 118.88, 118.55,
18.44, 112.64, 112.01, 109.77, 70.20, 62.77, 57.61, 46.42, 22.18. HRMS (ESI):
+
Calcd. for C H N NaO [M+Na] 476.1950; found 476.1988.
2
8
27
3
3
4
8
.1.3.6
-Benzyloxy-5-(2-hydroxy-1-((4-(4-phenylbutoxy)butyl)amino)ethyl)quinolin-2(1H)-
one (4f).
White solid; mp: 254.3-258.1°C; yield 30.2%; H-NMR (400 MHz, DMSO-d ) δ:
1
6
1
0.92 (s, 1H), 9.43 (s, 1H), 9.16 (s, 1H), 8.23 (d, J = 10.1 Hz, 1H), 7.59 (d, J = 7.1
Hz, 2H), 7.49 (d, J = 8.5 Hz, 1H), 7.38 (t, J = 7.3 Hz, 2H), 7.32 (dd, J = 7.9, 4.1 Hz,
2
H), 7.29 – 7.22 (m, 2H), 7.21 – 7.10 (m, 3H), 6.62 (d, J = 9.9 Hz, 1H), 5.55 (t, J =
.1 Hz, 1H), 5.34 (s, 2H), 4.89 (s, 1H), 3.81 (dt, J = 16.3, 11.3, 5.3 Hz, 2H), 3.26 (q, J
5

ACCEPTED MANUSCRIPT
=
6.4 Hz, 4H), 2.91 (s, 1H), 2.73 (s, 1H), 2.60 – 2.51 (m, 2H), 1.76 – 1.59 (m, 2H),
1
3
1
.57 – 1.47 (m, 2H), 1.47 – 1.32 (m, 4H). C-NMR (150 MHz, DMSO-d ) δ: 161.23,
6
1
44.78, 142.56, 136.93, 136.38, 130.09, 128.83, 128.74, 128.69, 128.43, 128.33,
26.11, 123.47, 121.23, 118.58, 112.61, 70.22, 69.77, 62.61, 57.57, 45.90, 35.34,
1
+
2
9.20, 28.04, 26.75, 23.24. HRMS (ESI): Calcd. for C H N O [M+H] 515.2910;
32
39
2
4
found 515.2945.
4
8
.1.3.7
-Benzyloxy-5-(2-hydroxy-1-((6-(4-phenylbutoxy)hexyl)amino)ethyl)quinolin-2(1H)-
one (4g).
White solid; mp: 199.9-201.2°C; yield 19.3%; H-NMR (600 MHz, DMSO-d ) δ:
1
6
1
0.87 (s, 1H), 9.46 (s, 1H), 9.14 (s, 1H), 8.23 (d, J = 10.1 Hz, 1H), 7.59 (d, J = 7.3
Hz, 2H), 7.50 (d, J = 8.5 Hz, 1H), 7.39 (t, J = 7.5 Hz, 2H), 7.32 (dd, J = 7.9, 4.6 Hz,
H), 7.26 (t, J = 7.5 Hz, 2H), 7.16 (t, J = 8.3 Hz, 3H), 6.62 (d, J = 9.9 Hz, 1H), 5.53
2
(
s, 1H), 5.34 (s, 2H), 4.90 (s, 1H), 3.97 – 3.82 (m, 1H), 3.78 (d, J = 11.1 Hz, 1H), 3.32
t, J = 6.5 Hz, 2H), 3.27 (t, J = 6.4 Hz, 2H), 2.89 (s, 1H), 2.70 (s, 1H), 2.56 (t, J = 7.6
(
Hz, 2H), 1.62 (d, J = 5.7 Hz, 2H), 1.57 (dt, J = 15.4, 7.5 Hz, 2H), 1.47 (dt, J = 13.3,
1
3
6
1
1
5
.5 Hz, 2H), 1.43 – 1.36 (m, 2H), 1.21 (s, 4H). C-NMR (150 MHz, DMSO-d ) δ:
6
61.13, 144.67, 142.52, 136.86, 136.34, 129.98, 128.74, 128.64, 128.60, 128.34,
28.25, 126.01, 123.56, 123.35, 121.21, 118.50, 112.54, 70.13, 70.10, 70.09, 62.56,
7.53, 45.83, 35.25, 29.25, 29.15, 28.04, 26.15, 25.61, 25.53. HRMS (ESI): Calcd. for
+
C H N O [M+H] 543.3223; found 543.3264.
34
43
2
4

ACCEPTED MANUSCRIPT
4
8
.1.3.8
-Benzyloxy-5-(2-hydroxy-1-((2-hydroxyphenethyl)amino)ethyl)quinolin-2(1H)-one
(4h).
1
White solid; mp: 240.2-241.4°C; yield 26.5%; H-NMR (600 MHz, DMSO-d ) δ:
6
1
0.82 (s, 1H), 9.64 (s, 1H), 8.91 (s, 1H), 8.31 (d, J = 10.0 Hz, 1H), 7.58 (d, J = 7.3
Hz, 2H), 7.47 (d, J = 7.3 Hz, 2H), 7.38 (t, J = 7.6 Hz, 2H), 7.32 (dd, J = 9.9, 5.1 Hz,
H), 7.28 – 7.18 (m, 5H), 7.07 (d, J = 8.1 Hz, 1H), 6.92 (t, J = 7.4 Hz, 1H), 6.59 (d, J
9.9 Hz, 1H), 6.07 (s, 1H), 5.50 (d, J = 10.0 Hz, 1H), 5.33 (s, 2H), 5.13 (s, 2H), 3.19
3
=
–
1
3
3.12 (m, 2H), 3.12 – 2.93 (m, 4H). C-NMR (150 MHz, DMSO-d ) δ: 161.21,
6
1
1
1
56.35, 144.78, 137.45, 136.88, 136.35, 130.52, 130.08, 128.86, 128.82, 128.71,
28.42, 128.29, 128.16, 127.70, 125.69, 123.45, 123.37, 121.20, 118.54, 112.65,
12.53, 70.20, 69.38, 62.74, 57.75,45.41, 27.29. HRMS (ESI): Calcd. for C H N O
4
33
33
2
+
[
M+H] 521.2440; found 521.2437.
4
8
.1.3.9
-Benzyloxy-5-(2-hydroxy-1-((3-hydroxyphenethyl)amino)ethyl)quinolin-2(1H)-one
(4i).
1
White solid; mp: 213.9-215.0°C; yield 16.8%; H-NMR (600 MHz, DMSO-d ) δ:
6
1
0.88 (s, 1H), 9.71 (s, 1H), 9.40 (s, 1H), 8.20 (d, J = 10.1 Hz, 1H), 7.59 (d, J = 7.4
Hz, 2H), 7.52 (d, J = 8.4 Hz, 1H), 7.42 (d, J = 7.3 Hz, 2H), 7.38 (td, J = 7.6, 2.3 Hz,
H), 7.35 – 7.29 (m, 3H), 7.20 (t, J = 7.9 Hz, 1H), 6.87 (dd, J = 8.2, 2.3 Hz, 1H), 6.81
s, 1H), 6.73 (d, J = 7.6 Hz, 1H), 6.62 (d, J = 9.9 Hz, 1H), 5.59 (s, 1H), 5.34 (s, 2H),
4
(

ACCEPTED MANUSCRIPT
5
.05 (s, 2H), 4.94 (s, 1H), 3.88 (d, J = 3.0 Hz, 1H), 3.81 (d, J = 10.1 Hz, 1H), 3.20 (d,
1
3
J = 5.2 Hz, 1H), 3.05 – 2.96 (m, 2H), 2.93 (dd, J = 11.1, 7.9 Hz, 1H). C-NMR (150
MHz, DMSO-d ) δ: 161.14, 158.89, 144.69, 139.22, 137.36, 136.85, 136.36, 130.10,
6
1
1
29.98, 128.78, 128.74, 128.33, 128.26, 128.20, 128.06, 123.55, 123.30, 121.28,
18.42, 115.50, 113.30, 112.58, 70.14, 69.48, 62.54, 57.71, 46.89, 31.91. HRMS
+
(
ESI): Calcd. for C H N O [M+H] 521.2440; found 521.2445.
3
3
33
2
4
4
8
.1.3.10
-Benzyloxy-5-(2-hydroxy-1-((4-hydroxyphenethyl)amino)ethyl)quinolin-2(1H)-one
(4j).
1
White solid; mp: 178.0-179.3°C; yield 23.4%; H-NMR (600 MHz, DMSO-d ) δ:
6
1
0.87 (s, 1H), 9.66 (s, 1H), 9.34 (s, 1H), 8.20 (d, J = 10.1 Hz, 1H), 7.59 (d, J = 7.4
Hz, 2H), 7.51 (d, J = 8.4 Hz, 1H), 7.42 (d, J = 7.3 Hz, 2H), 7.38 (dt, J = 7.6, 5.8 Hz,
H), 7.32 (t, J = 8.1 Hz, 3H), 7.06 (d, J = 8.6 Hz, 2H), 6.92 (d, J = 8.6 Hz, 2H), 6.62
d, J = 9.9 Hz, 1H), 5.58 (s, 1H), 5.35 (s, 2H), 5.05 (s, 2H), 4.94 (s, 1H), 3.94 – 3.84
4
(
(
1
3
m, 1H), 3.80 (d, J = 11.5 Hz, 1H), 3.14 (s, 1H), 3.02 – 2.81 (m, 3H). C-NMR (150
MHz, DMSO-d ) δ: 161.13, 157.53, 144.67, 137.47, 136.86, 136.37, 129.92, 129.69,
6
1
1
28.79, 128.34, 128.27, 128.16, 127.98, 123.58, 123.31, 121.29, 118.43,
15.33,112.59, 70.14, 69.52, 62.53, 57.65, 47.27, 31.08. HRMS (ESI): Calcd. for
+
C H N NaO [M+Na] 543.2260; found 543.2203.
33
32
2
4
4
8
.1.3.11
-Benzyloxy-5-(2-hydroxy-1-((2-methoxyphenethyl)amino)ethyl)quinolin-2(1H)-one
(4k).

ACCEPTED MANUSCRIPT
1
White solid; mp: 234.3-236.4°C; yield 24.5%; H-NMR (600 MHz, DMSO
-d6) δ:
1
0.90 (s, 1H), 9.69 (s, 1H), 9.40 (s, 1H), 8.22 (d, J = 10.0 Hz, 1H), 7.60 (d, J = 7.5
Hz, 2H), 7.53 (d, J = 8.1 Hz, 1H), 7.39 (t, J = 7.4 Hz, 2H), 7.32 (dd, J = 14.8, 7.8 Hz,
H), 7.21 (t, J = 7.7 Hz, 1H), 7.07 (d, J = 7.2 Hz, 1H), 6.90 (d, J = 8.2 Hz, 1H), 6.86
t, J = 7.4 Hz, 1H), 6.62 (d, J = 9.9 Hz, 1H), 5.59 (s, 1H), 5.36 (s, 2H), 4.97 (s, 1H),
2
(
3
.86 (s, 1H), 3.80 (d, J = 10.7 Hz, 1H), 3.60 (s, 3H), 3.07 (s, 1H), 3.02 – 2.87 (m, 3H).
1
3
C-NMR (150 MHz, DMSO-d ) δ: 161.25, 157.43, 14.72, 136.97, 136.48, 130.38,
6
1
1
30.08, 128.83, 128.42, 128.33, 125.34, 123.51, 123.40, 121.28, 120.97, 118.58,
12.62, 111.33, 70.14, 62.73, 57.62, 55.0, 45.40, 27.08. HRMS (ESI): Calcd. for
+
C H N O [M+H] 445.2127; found 445.2157.
27
29
2
4
4
8
.1.3.12
-Benzyloxy-5-(2-hydroxy-1-((3-methoxyphenethyl)amino)ethyl)quinolin-2(1H)-one
(
4l).
White solid; mp: 242.9-243.6°C; yield 25.4%; H-NMR (600 MHz, DMSO
0.88 (s, 1H), 9.58 (d, J = 99.1 Hz, 1H), 9.28 (d, J = 73.6 Hz, 1H), 8.19 (d, J = 10.1
Hz, 1H), 7.59 (d, J = 7.3 Hz, 2H), 7.47 (dd, J = 34.3, 9.2 Hz, 1H), 7.38 (t, J = 7.5 Hz,
H), 7.32 (t, J = 8.3 Hz, 2H), 7.20 (t, J = 8.8 Hz, 1H), 6.79 (d, J = 7.9 Hz, 1H), 6.71
1
-d6) δ:
1
2
(d, J = 6.9 Hz, 2H), 6.62 (d, J = 9.8 Hz, 1H), 5.59 (s, 1H), 5.34 (s, 2H), 5.08 – 4.81
(m, 1H), 4.02 – 3.77 (m, 2H), 3.71 (s, 3H), 3.19 (s, 1H), 3.09 – 2.84 (m, 3H).
1
3
C-NMR (150 MHz, DMSO-d ) δ: 161.34, 159.86, 144.78, 139.26, 136.94, 136.45,
6
1
1
30.17, 130.07, 128.83, 128.43, 128.35, 123.64, 123.41, 121.36, 121.11, 118.50,
14.66, 112.67, 112.60, 70.20, 62.57, 57.73, 55.43, 47.03, 32.02. HRMS (ESI):

ACCEPTED MANUSCRIPT
+
Calcd. for C H N O [M+H] 445.2127; found 445.2166.
2
7
29
2
4
4
8
.1.3.13
-Benzyloxy-5-(2-hydroxy-1-((4-methoxyphenethyl)amino)ethyl)quinolin-2(1H)-one
(
4m).
White solid; mp: 191.8-192.9°C; yield 21.1%; H-NMR (400 MHz, DMSO
0.92 (s, 1H), 9.70 (s, 1H), 9.39 (s, 1H), 8.21 (d, J = 10.1 Hz, 1H), 7.60 (d, J = 8.3
Hz, 2H), 7.53 (d, J = 8.5 Hz, 1H), 7.39 (t, J = 7.3 Hz, 2H), 7.32 (t, J = 8.2 Hz, 2H),
1
-d6) δ:
1
7
1
1
.06 (d, J = 8.7 Hz, 2H), 6.84 (d, J = 8.7 Hz, 2H), 6.62 (d, J = 9.9 Hz, 1H), 5.60 (s,
H), 5.35 (s, 2H), 5.04 – 4.87 (m, 1H), 3.89 (dd, J = 11.1, 6.3 Hz, 1H), 3.80 (dd, J =
1
3
1.5, 4.5 Hz, 1H), 3.70 (s, 3H), 3.14 (s, 1H), 2.91 (dt, J = 11.3, 7.8 Hz, 3H). C-NMR
(
150 MHz, DMSO-d ) δ: 161.23, 158.53, 144.77, 136.94, 136.45, 130.06, 129.48,
6
1
28.83, 128.43, 128.36, 123.65, 123.40, 121.37, 118.51, 114.50, 112.67, 70.21, 62.60,
+
5
7.71, 55.49, 57.40, 31.16. HRMS (ESI): Calcd. for C H N O [M+H] 445.2127;
27
29
2
4
found 445.2196.
4
8
.1.3.14
-Benzyloxy-5-(1-((2-chlorophenethyl)amino)-2-hydroxyethyl)quinolin-2(1H)-one
(
4n).
White solid; mp: 253.5-255.5°C; yield 20.1%; H-NMR (400 MHz, DMSO
0.94 (s, 1H), 9.88 (s, 1H), 9.56 (s, 1H), 8.24 (d, J = 10.1 Hz, 1H), 7.60 (d, J = 7.1
Hz, 2H), 7.55 (d, J = 8.5 Hz, 1H), 7.43 – 7.20 (m, 8H), 6.63 (d, J = 9.9 Hz, 1H), 5.63
1
-d6) δ:
1
(s, 1H), 5.36 (s, 2H), 5.00 (s, 1H), 3.88 (dd, J = 11.4, 5.7 Hz, 1H), 3.85 – 3.71 (m,
1
3
1
H), 3.24 – 3.05 (m, 3H), 2.97 (s, 1H). C-NMR (150 MHz, DMSO-d ) δ: 160.17,
6

ACCEPTED MANUSCRIPT
1
1
4
4
4
8
43.67, 135.86, 135.41, 134.13, 132.29, 130.28, 129.03, 128.81, 128.27, 127.75,
27.34, 127.27, 127.03, 122.41, 122.34, 120.25, 117.47, 111.61, 69.10, 61.65, 56.70,
+
4.11, 28.80. HRMS (ESI): Calcd. for C H ClN NaO [M+Na] 471.1451; found
2
6
25
2
3
71.1488.
.1.3.15
-Benzyloxy-5-(1-((3-chlorophenethyl)amino)-2-hydroxyethyl)quinolin-2(1H)-one
4o).
White solid; mp: 236.1-237.3°C; yield 18.3%; H-NMR (400 MHz, DMSO
0.91 (s, 1H), 9.74 (s, 1H), 9.40 (s, 1H), 8.20 (d, J = 10.1 Hz, 1H), 7.60 (d, J = 7.1
Hz, 2H), 7.51 (d, J = 8.5 Hz, 1H), 7.39 (t, J = 7.3 Hz, 2H), 7.35 – 7.29 (m, 4H), 7.27
s, 1H), 7.14 (d, J = 6.9 Hz, 1H), 6.63 (d, J = 9.9 Hz, 1H), 5.60 (s, 1H), 5.35 (s, 2H),
(
1
-d6) δ:
1
(
5
.07 – 4.89 (m, 1H), 3.88 (dd, J = 11.3, 6.2 Hz, 1H), 3.81 (dd, J = 11.5, 4.5 Hz, 1H),
1
3
3
.23 (dd, J = 17.1, 8.4 Hz, 1H), 3.09 – 2.90 (m, 3H). C-NMR (150 MHz, DMSO-d )
6
δ: 160.16, 143.69, 139.25, 135.87, 135.38, 132.52, 129.84, 128.99, 127.83, 127.76,
1
27.35, 127.28, 126.75, 126.13, 122.58, 122.31, 120.30, 117.42, 111.57, 69.12, 61.51,
+
5
6.76, 45.68, 30.48. HRMS (ESI): Calcd. for C H ClN NaO [M+Na] 471.1451;
2
6
25
2
3
found 471.1489.
4
8
.1.3.16
-Benzyloxy-5-(1-((4-chlorophenethyl)amino)-2-hydroxyethyl)quinolin-2(1H)-one
(
4p).
White solid; mp: 205.1-206.8°C; yield 19.6%; H-NMR (400 MHz, DMSO
0.91 (s, 1H), 9.78 (s, 1H), 9.44 (s, 1H), 8.21 (d, J = 10.1 Hz, 1H), 7.60 (d, J = 7.2
1
-d6) δ:
1

ACCEPTED MANUSCRIPT
Hz, 2H), 7.53 (d, J = 8.5 Hz, 1H), 7.47 – 7.29 (m, 7H), 7.19 (d, J = 8.4 Hz, 2H), 6.62
(d, J = 9.9 Hz, 1H), 5.60 (s, 1H), 5.35 (s, 2H), 4.94 (s, 1H), 3.88 (s, 1H), 3.82 (s, 1H),
1
3
3
1
1
.20 (s, 1H), 3.12 – 2.86 (m, 3H). C-NMR (150 MHz, DMSO-d ) δ: 161.24, 144.78,
6
36.95, 136.75, 136.44, 131.86, 130.93, 130.07, 129.01, 128.84, 128.44, 128.36,
23.61, 123.41, 121.35, 118.49, 112.66, 70.20, 62.56, 57.79, 46.88, 31.31. HRMS
+
(
ESI): Calcd. for C H ClN NaO [M+Na] 471.1451; found 471.1488.
2
6
25
2
3
4
8
.1.3.17
-Benzyloxy-5-(2-hydroxy-1-((2-methyl-1-phenylpropan-2-yl)amino)ethyl)quinolin-2
(
1H)-one (4q).
White solid; mp: 254.3-255.8°C; yield 18.1%; H-NMR (600 MHz, DMSO
0.88 (s, 1H), 9.36 (s, 1H), 8.84 (s, 1H), 8.42 (d, J = 9.9 Hz, 1H), 7.65 (s, 1H), 7.60
d, J = 7.3 Hz, 2H), 7.38 (t, J = 7.5 Hz, 2H), 7.32 (dt, J = 10.7, 9.7 Hz, 4H), 7.26 (t, J
7.2 Hz, 1H), 7.15 (d, J = 7.0 Hz, 2H), 6.65 (d, J = 9.9 Hz, 1H), 5.59 (s, 1H), 5.35 (s,
H), 5.11 (s, 1H), 3.93 (s, 1H), 3.87 (s, 1H), 3.05 (d, J = 12.6 Hz, 1H), 2.95 (d, J =
1
-d6) δ:
1
(
=
2
1
1
1
4
4
4
8
1
3
0.7 Hz, 1H), 1.12 (d, J = 6.6 Hz, 6H). C-NMR (150 MHz, DMSO-d ) δ: 160.12,
6
43.68, 135.87, 135.30, 134.64, 130.05, 128.93, 127.76, 127.64, 127.35, 127.29,
26.39, 126.37, 124.41, 122.59, 121.35, 116.92, 111.50, 69.16, 63.08, 61.24, 52.97,
+
2.95, 22.94, 22.49. HRMS (ESI): Calcd. for C H N O [M+H] 443.2335; found
2
8
31
2
3
43.2367.
.1.3.18
-Benzyloxy-5-(1-((2,2-diphenylethyl)amino)-2-hydroxyethyl)quinolin-2(1H)-one
4r)
(

ACCEPTED MANUSCRIPT
1
White solid; mp: 258.4-260.7°C; yield 19.9%; H-NMR (150 MHz, DMSO
-d6) δ:
1
–
0.79 (s, 1H), 10.56 (s, 1H), 9.42 (s, 1H), 8.95 (s, 1H), 8.02 (d, J = 10.0 Hz, 1H), 7.35
7.14 (m, 1H), 7.05 (d, J = 8.3 Hz, 1H), 6.54 (d, J = 9.9 Hz, 1H), 5.57 (s, 1H), 4.89
13
(
s, 1H), 4.59 (t, J = 6.7 Hz, 1H), 3.91 (s, 1H), 3.72 (s, 2H), 3.55 (s, 1H). C-NMR
(
150 MHz, DMSO-d ) δ: 161.25, 144.82, 141.77, 141.29, 136.41, 130.08, 129.25,
6
1
1
29.20, 128.84, 128.44, 128.36, 128.29, 128.25, 128.15, 128.14, 127.51, 127.42,
23.33, 123.28, 121.20, 118.77, 112.65, 70.19, 62.55, 58.53, 49.76, 47.78. HRMS
+
(
ESI): Calcd. for C H N O [M+H] 491.2335; found 491.2371.
3
2
31
2
3
4
5
.1.3.19
-(1-((4-Aminophenethyl)amino)-2-hydroxyethyl)-8-(benzyloxy)quinolin-2(1H)-one
(4s).
White
solid;
mp:
161.1-162.5°C;
yield
20.3%;
1
H-NMR (600 MHz, DMSO-d ) δ: 10.90 (s, 1H), 9.52 (s, 1H), 9.23 (s, 1H), 8.19 (d, J
6
=
=
=
=
=
=
10.1 Hz, 2H), 7.59 (d,
7.5 Hz, 2H), 7.32 (t,
9.9 Hz, 1H), 6.50 (d,
J
= 7.4 Hz, 2H), 7.47 (d,
= 7.9 Hz, 2H), 6.79 (d,
J
= 8.5 Hz, 1H), 7.39 (t,
= 8.2 Hz, 2H), 6.62 (d,
J
J
J
J
J
= 8.2 Hz, 2H), 5.57 (t,
J
5.0 Hz, 1H), 5.35 (s, 2H), 5.23 (s, 1H), 4.92 (s, 1H), 3.85 (dt,
11.3, 5.7 Hz, 1H), 3.82 – 3.75 (m, 1H), 3.07 (s, 1H), 2.87 (d,
10.7 Hz, 1H), 2.85 – 2.78 (m, 1H), 2.78 – 2.69 (m, 1H).
J
J
1
3
C-NMR (150
MHz, DMSO-d ) δ: 161.21, 144.85, 136.93, 136.48, 130.15, 129.40, 128.83, 128.45,
6
1
3
28.32, 123.78, 123.42, 121.31, 118.54, 114.87, 112.75, 70.26, 62.59, 57.62, 47.64,
+
1.35. HRMS (ESI): Calcd. for C H N O [M+H] 430.2131; found 430.2135.
2
6
28
3
3

ACCEPTED MANUSCRIPT
4
.1.3.20
N-(4-(2-((1-(8-(Benzyloxy)-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethyl)amino)e
thyl)phenyl)benzamide (4t).
1
White solid; mp: 136.0-137.2°C; yield 13.7%; H-NMR (600 MHz, DMSO-d ) δ:
6
1
0.90 (s, 1H), 10.26 (s, 1H), 9.71 (d, J = 5.0 Hz, 1H), 9.39 (d, J = 7.7 Hz, 1H), 8.21 (d,
J = 10.1 Hz, 1H), 8.01 – 7.91 (m, 2H), 7.71 (d, J = 8.5 Hz, 2H), 7.59 (dd, J = 10.3, 7.5
Hz, 3H), 7.56 – 7.44 (m, 3H), 7.39 (t, J = 7.5 Hz, 2H), 7.36 – 7.27 (m, 2H), 7.13 (d, J
=
8.5 Hz, 2H), 6.63 (d, J = 9.9 Hz, 1H), 5.61 (s, 1H), 5.35 (s, 2H), 3.89 (dd, J = 11.7,
6
2
1
1
4
5
.3 Hz, 1H), 3.82 (dd, J = 11.7, 4.7 Hz, 1H), 3.24 – 3.18 (m, 1H), 2.96 (ddd, J = 32.4,
1
3
3.8, 14.6 Hz, 3H). C-NMR (150 MHz, DMSO-d ) δ: 165.90, 161.27, 144.84,
6
38.33, 136.95, 136.52, 135.38, 132.84, 132.07, 130.13, 129.17, 128.84, 128.49,
28.34, 128.11, 123.75, 123.47, 121.32, 121.00, 118.52, 112.74, 70.22, 62.68, 56.54,
+
7.23, 31.56. HRMS (ESI): Calcd. forC H N O [M+H] 534.2393; found
3
3
32
3
4
34.2389.
4
.1.3.21
N-(4-(2-((1-(8-(Benzyloxy)-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethyl)amino)e
thyl)phenyl)-2-phenylacetamide (4u).
1
White solid; mp: 185.5-186.9°C; yield 17.2%; H-NMR (600 MHz, DMSO-d ) δ:
6
1
0.88 (s, 1H), 10.24 (s, 1H), 9.52 (d, J = 4.3 Hz, 1H), 9.22 (d, J = 7.8 Hz, 1H), 8.23 (d,