Synthesis and biological evaluation of novel 2-oxo-1,2-dihydroquinoline-4-carboxamide derivatives for the treatment of esophageal squamous cell carcinoma

Article abstract of DOI:10.1016/j.ejmech.2018.05.042

No new and effective treatments have been approved for the treatment of esophageal squamous cell carcinoma (ESCC) in the past decade. Cisplatin and 5-fluoruracil are the most commonly used drugs for this disease. In order to develop a new class of drugs effective in our ESCC phenotypic screens, we began a systematic approach to generate novel compounds based on the 2-oxo-1,2-dihydroquinoline-4-carboxamide fragment. Herein, we report on the synthesis and initial assessment of 55 new analogues in two ESCC cell lines. Some of the active analogues with IC₅₀ values around 10 μM were tested in three additional cell lines. Our structure-activity relationships revealed remarkable alterations in the anti proliferative activities upon modest chemical modifications and autophagy modulation is a suggested mechanism of action.

Full text of DOI:10.1016/j.ejmech.2018.05.042

Accepted Manuscript
Synthesis and biological evaluation of novel 2-oxo-1,2-dihydroquinoline-4-
carboxamide derivatives for the treatment of esophageal squamous cell carcinoma
Mai I. Shahin, Joyeeta Roy, Maha Hanafi, Dongyao Wang, Urarika Luesakul, Yifeng
Chai, Nongnuj Muangsin, Deena S. Lasheen, Dalal A. Abou El Ella, Khaled A.
Abouzid, Nouri Neamati
PII:
S0223-5234(18)30455-0
10.1016/j.ejmech.2018.05.042
EJMECH 10453
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 9 April 2018
Revised Date: 21 May 2018
Accepted Date: 25 May 2018
Please cite this article as: M.I. Shahin, J. Roy, M. Hanafi, D. Wang, U. Luesakul, Y. Chai, N. Muangsin,
D.S. Lasheen, D.A. Abou El Ella, K.A. Abouzid, N. Neamati, Synthesis and biological evaluation of novel
2-oxo-1,2-dihydroquinoline-4-carboxamide derivatives for the treatment of esophageal squamous cell
carcinoma, European Journal of Medicinal Chemistry (2018), doi: 10.1016/j.ejmech.2018.05.042.
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ACCEPTED MANUSCRIPT
R
N
H
Br
O
Br
X
N
O
Br
N
O
F
NH
O
R
O
R
Series A
Series B
R= OH, Benzyl
R2
H
N
H
1
O
R1
X
N
O
2
4
O
F
NH
O
F
NH
Y
Y
X= C, N
Y=H, F
R2= OH, OCH3
R1= Cl, F, OCH3, CF3, OH, Ar, Alk
Series B and C

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Synthesis and biological evaluation of novel 2-oxo-1,2-
dihydroquinoline-4-carboxamide
derivatives
for
the
treatment of esophageal squamous cell carcinoma
[a,c]
Mai I. Shahin,[a,b] Joyeeta Roy,[a] Maha Hanafi,
Dongyao Wang,[a,d] Urarika
Luesakul,[a,e] Yifeng Chai,[d] Nongnuj Muangsin,[e] Deena S. Lasheen,[b] Dalal A.
Abou El Ella,[b,f] Khaled A. Abouzid,[b] and Nouri Neamati*[a]
[a] Department of Medicinal Chemistry, College of Pharmacy, University of Michigan, North Campus Research
Complex, 1600 Huron parkway, Ann Arbor, Michigan 48109, United States
[b] Pharmaceutical Chemistry Department, Faculty of Pharmacy, Ain Shams University, Abbassia, Cairo 11566,
Egypt
[c] Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt
[d] School of Pharmacy, Second Military Medical University, Shanghai, China, 200433
[e] Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand
[f] Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Nahda University, 62511 Beni Suef, Egypt
Abstract
No new and effective treatments have been approved for the treatment of esophageal squamous
cell carcinoma (ESCC) in the past decade. Cisplatin and 5-fluoruracil are the most commonly
used drugs for this disease. In order to develop a new class of drugs effective in our ESCC
phenotypic screens, we began a systematic approach to generate novel compounds based on the
2-oxo-1,2-dihydroquinoline-4-carboxamide fragment. Herein, we report on the synthesis and
initial assessment of 55 new analogues in two ESCC cell lines. Some of the active analogues
with IC50 values around 10 µM were tested in three additional cell lines. Our structure-activity
relationships revealed remarkable alterations in the anti proliferative activities upon modest
chemical modifications and autophagy modulation is a suggested mechanism of action.

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Keywords
Quinoline-4-carboxamide; esophageal squamous cell cancer; autophagy; structure-activity
relationships; chloroquine.
1.
Introduction
The two predominant histological types of esophageal cancers are esophageal squamous cell
cancer (ESCC) and adenocarcinoma (AC) [1-3]. These cancers are different in terms of
epidemiology, pathogenesis, tumor biology, and prognosis [4-9]. Recent studies have
demonstrated striking similarities between ESCC and head and neck cancers [10]. From
>400,000 deaths from esophageal cancer in the world each year, 80% are due to ESCC [11-13].
ESCC is an aggressive disease with poor prognosis and is typically diagnosed in late stage of the
disease. ESCC is the fourth leading cause of cancer death in the United States with a 5-year
survival rate of 19%. The standard of care treatment consists of the classical modalities including
surgery, radiotherapy, and the combination of two old drugs, cisplatin and 5-FU [2],[14-16].
Unfortunately, only 20–40% of patients show significant response. Therefore, novel drugs
targeting key genes or pathways implicated in ESCC are needed to effectively treat this disease.
In recent years, neoadjuvant chemotherapy followed by surgery have been adopted as a first-line
treatment for advanced ESCC tumor [17]. Selected small molecule drugs with different
biological targets that are currently under clinical development are listed in Table 1. Despite all
these substantial efforts, no significant improvements have been observed in the overall survival
rates of ESCC patients. Therefore, there is an unmet need for developing new effective
therapeutic agents for ESCC.

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Table1. Selected examples of small molecule drugs under clinical trials for ESCC treatment
Mode of
action
Trial
stage
Agent
Structure
Tumor type
Trial number
References
Hedgehog Early
Esophageal
cancer
Itraconazole
pathway
phase
I
NCT02749513
[18]
inhibition
Ganetespib
(STA-9090)
HSP90
inhibition
Phase
I
Esophageal
cancer
NCT02389751
[19]
Taladegib
Hedgehog
pathway
inhibition
Phase
I/II
Esophageal
cancer
NCT02530437
NCT0213892
NCT00917462
[20]
[20]
[21]
(LY2940680
)
Hedgehog
pathway
inhibition
Sonidegib
(LDE225)
Phase
I
Esophageal
cancer
Esophageal
cancer
Gastroesophageal
Junction Cancer
Gastric
VEGFR
inhibition
Phase
II
Sorafenib
Adenocarcinoma
Gastroesophageal
Junction
Adenocarcinoma
Recurrent
Epacadostat
(INCB024360)
IDO1
inhibition
Phase
II
Esophageal
Carcinoma
NCT03196232
[22]
Recurrent Gastric
Carcinoma
Unresectable
Esophageal
Carcinoma
Autophagy, an evolutionarily conserved cellular response, mitigates stresses through catabolic
degradation of intracellular components to maintain hemostasis. The impact of autophagy is
investigated in many diseases especially cancer [23, 24]. Its dual role in cancer suggests that
autophagy modulation through inhibition or activation can be an effective strategy for treating
cancer [25]. Despite its complex role in cancer, many clinical interventions are thought to be

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through autophagy inhibition [26]. The solely clinically available drugs suppressing autophagy
are chloroquine and hydroxychloroquine. Chloroquine in combination with various drugs has
been explored as an effective strategy to treat different types of cancers including ESCC [27].
The mechanistic studies revealed that chloroquine exerts its anti-tumor activity mainly through
autophagy inhibition [28, 29]. A recent study reported that autophagy inhibition sensitizes
esophageal cancer cells to radiation [30].
In this context, 2-oxo-1,2-dihydroquinoline-4-carboxamide fragment was used as a core scaffold
to synthesize three series of compounds consisting of 55 derivatives. They were evaluated for
their inhibition of cell proliferation in two representative ESCC cell lines, KYSE410 and
KYSE70. We used the 2-oxo-1,2-dihydroquinoline-4-carboxamide fragment for synthetic
elaboration to design novel compounds for the following reasons: 1. This fragment is drug like
(MW = 264, tPSA = 58, cLogP = 1.3, H-bond donor= 2, H-bond acceptor = 2). 2. This fragment
is novel because it is not present in any clinically used drug. 3. 2-Oxo-1,2-dihydroquinoline-4-
carboxamide can be readily synthesized from commercially available isatin derivatives and
malonic acid. 4. This fragment is amenable to facile modifications producing a large library of
novel analogues with desirable pharmacokinetics and pharmaceutics properties. The stepwise
optimization strategy presented in this study yielded compounds with novel mechanisms to treat
ESCC. Therefore, the optimized compounds are suitable for further developments paving the
way for designing second-generation compounds for in vivo studies.
2.
Chemistry
Series A, B, and C were synthesized according to Schemes 1 and 2a,b. 7-Bromoquinoline-2,4-
dicarboxylic acid (1) and 7-substituted-2-oxo-1,2-dihydroquinoline-4-carboxylic acid (5a, 5c, 5d

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and 5e) were synthesized by reacting the commercially available 6-substituted isatin with
pyruvic acid sodium salt and malonic acid respectively as reported [31, 32].
Similar synthetic method was applied to prepare compound 14 from the commercially available
7-methoxyindoline-2,3-dione. 6-Methoxy-1H-pyrrolo[2,3-b]pyridine-2,3-dione
(4) was
synthesized by Pyridinium chlorochromate mediated oxidation of 6-methoxy-1H-pyrrolo[2,3-
b]pyridine [33]. Decarboxylation at position 2 of compound 1 was achieved by refluxing in
nitrobenzene as reported [34]. The commercially available 7-fluoro-2-oxo-1,2-dihydroquinoline-
4-carboxylic acid (5b) and the synthesized carboxylic acids 2, 5a, 5c, 5d, 5e and 5f were reacted
with the corresponding amines to give the targeted compounds (Schemes 1 and 2a, b; Series A
and B). BBr3-mediated demethylation of 6x and 15 afforded the corresponding hydroxy
analogs 6y and 16 [35]. To replace the bromo atom with the acetylene moiety,
bis(triphenylphosphine)palladium chloride was used as
a
catalyst to couple of
trimethylsilylacetylene 6i. Compound 8 was further deprotected to yield the free acetylenic
derivative (9) [36, 37]. Compound 6i was benzylated using benzyl chloride and potassium
carbonate in DMF to yield 10 [38].

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Table2. Cytotoxicity ± SD of compounds 3a-d and cisplatin in two ESCC cell lines
Coupling the bromo-derivatives with various boronic acid under Suzuki–Miyaura cross-coupling
conditions afforded series C analogues. This was achieved by refluxing the bromo-derivative and
the boronic acid in dioxane: water (4:1) using cesium carbonate as base and [1,1'-
bis(diphenylphosphino)ferrocene]palladium(II) dichloride (Pd(dppf)Cl2) as the palladium source
[39]. The N,N-diethylpentane-1,4-diamine moiety was appended to the corresponding bromo
derivatives 6i and 12 under conditions of Buchwald Hartwig amination. This coupling afforded
derivatives 11 and 13 using Xphos: (2-Dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl) and
Pd2(dba)3: (Tris(dibenzylideneacetone)dipalladium(0)) as catalysts in the presence of sodium
tert-butoxide as a base [40]. Compound 18 was synthesized according to the pathway outlined in
Scheme 3. Chlorination of N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide using
POCl3 resulted in 17 that was further oxidized using mCPBA (meta-chloroperoxybenzoic acid)

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to yield the N-oxide intermediate. This intermediate was directly converted to the targeted
compound 18 using 10% aq. HCl under reflux condition [41].

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Table 3. Cytotoxicity ± SD of compounds 6a-z, 13, 15 and 16 in two ESCC cell lines

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Table 4. Cytotoxicity ± SD of compounds 7a-r, 9, 10, 11and 18 in two ESCC cell lines

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3.
Results and discussion
3.1. Lead optimization and SAR analysis
To design compounds containing the 2-oxo-1,2-dihydroquinoline-4-carboxamide fragment,
various analogues were synthesized through stepwise structure optimization process. MTT assay
was performed to assess the antiproliferative potency of all compounds in KYSE410 and
KYSE70 cell lines (Tables 2, 3 and 4). Cisplatin, a clinically used drug was used as control. As
shown in Fig. 1, four positions were explored by applying different modifications. First, series A
and B were synthesized in parallel with different aliphatic and aromatic fragments at position 4.
Compounds 6i was the most active with IC50 values 7.3 ± 1.1 and 8.9 ± 0.1 µM, respectively in
KYSE410 and KYSE70 cell lines. Having another fluorine atom in the other meta position in
compound 6r maintained similar IC50 values around 10 µM in both cell lines. This illustrates the
importance of having a fluorine atom at a meta position of the phenyl ring. Other derivatives
bearing different substituents on this phenyl ring on different positions did not produce potent
compounds (e.g. compound 6o which has a tertiary butyl group on the para position). The dose
response curve and the colony formation assay of the active compound 6r and the inactive
compound 6o in comparison to cisplatin is shown in Fig. 2.

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Fig. 1. Lead optimization paradigm
The replacement of the bromo atom at position 7 with the fluoro, chloro, trifloromethyl,
methoxy, hydroxyl, vinyl and acetylene groups negatively affects the activity. Also, neither the
bioisosteric replacement of quinolone with naphthyridine nor shifting the substitution to be at
position 8 instead of position 7 showed improvement in the activities. Since 6i and 6r are
acceptable starting point for optimization, further modifications were carried out as follow: Step
1: Position 1 modifications: We synthesized compounds 10 and 18 to test the effect of
hydrophilic and hydrophobic moieties at Position 1. Compound 10, with a hydrophobic group
showed diminished activity. Compound 18 bearing a hydrophilic hydroxyl group at position 1

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did not improve activity. This suggests the importance of having 2-oxo-1,2-dihydroquinoline
skeleton without any modifications at position 1. Step 2: Position 7 optimization: Starting from
compound 6i, further optimization was carried out on position 7. Series C (compounds 7a-r)
were synthesized using Suzuki-Miyaura cross coupling reaction where the bromo atom was
replaced with different aromatic and aliphatic rings. As illustrated in Table 4, the para fluoro
substituted phenyl ring (compound 7c) showed moderate potency where the activity decreased
by moving this fluoro atom to meta then to ortho positions. Similar cytotoxicity profile was
observed by introducing a pyran and a 2-fluoropyridine fragments on compounds 7m and 7o,
respectively. Compound 7f having a thiophene ring showed IC50 values of 4.3 and 5.1 µM in
KYSE410 and KYSE70, respectively.
Fig. 2. Compounds with a meta fluoro substitutions are active. (A) and (B) Compound 6r but not 6o is more
potent than cisplatin in KYSE410 and KYSE70 cells, respectively. (C) and (D) Compound 6r completely block
formation of colonies at doses above 3 µM in KYSE410 and KYSE70 cells.

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Step 3: Adding the N,N-diethylpentane-1,4-diamine moiety: It was important to synthesize
examples of compounds having this feature to test the effect on both cytotoxicity and autophagy
process. That’s why we synthesized compounds 11 and 13 having this group in two variant
positions. Despite the drop in the cytotoxicity, they behaved like chloroquine in its inhibitory
activity of autophagy pathway as will be discussed in the western blot results.
From all the synthesized compounds, the active compounds (6f, 6i, 6r, 7c, 7f and 7m) were
further tested in pancreatic cancer MIA PaCa-2 and two ovarian cancer OVCAR8 and SKOV3
cell lines. Compound 7f showed best cytotoxic activity over the other compounds that only
showed moderate cytotoxicity against them. (Table 5).
Table 5. IC50 ± SD (µM) values of compounds 6f, 6i, 6r, 7c, 7f and 7m in KYSE410, KYSE70, MIA PaCa-2, OVCAR8
and SKOV3 cell lines using MTT assay.
Compound KYSE410 KYSE70 MIA PaCa-2 OVCAR8 SKOV3
9.8 ±3.5 11.0 ± 0.3 21.3 ± 6.0
>20
>20
6f
6i
7.3 ± 1.1 8.9 ± 0.1
11.2 ± 2.2 10.7 ± 0.1
10.3 ± 1.1 3.3 ± 0.1
7.5 ± 3.6
7.8 ± 1.2
5.8 ± 1.1 10.6 ± 3.8
12.1 ± 4.6 10.8 ± 1.7
6r
7c
7f
19.3 ± 2.5 22.3 ± 2.6 16.6 ± 2.2
4.3 ±2.2
5.1 ± 2.8
4.5 ± 1.6
8.8 ± 1.6
13.4 ± 4.4 6.7 ± 2.0
21.8 ± 0.8 13.3 ± 4.6
12.1 ± 2.2 10.5 ± 0.6
7m
3.2. Lead compounds decrease bafilomycin-induced LC3B levels
To further investigate the mechanism of action of the lead compounds, the levels of LC3B, a
marker of autophagy, were measured upon treatment with compounds 6i and 6r. The experiment
was carried out in the presence and absence of bafilomycin-A1, a lysosome inhibitor.
Bafilomycin-A1 inhibits autophagy at late stages and results in increased levels of LC3B. The

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autophagy inhibitor chloroquine was used as a control. In contrast to chloroquine, compounds 6i
and 6r cause a decrease in the bafilomycin-induced LC3B levels after 24h treatment (Fig. 3A
and 3B). To confirm these results, we repeated the experiment using different concentrations of
compound 6r. The Western blot analysis showed a concentration-dependent decrease in the
bafilomycin-induced LC3B levels (Fig. 3C). Furthermore, we tested whether proteasome
inhibition will block the effect of 6r. We pre-treated the cells with the proteasome inhibitor
MG132 for 1h prior to a 24h treatment with 6r. The results showed no decrease in LC3B levels
in the presence of MG132 suggesting that the effect of 6r on LC3B levels are mediated by the
proteasome. In conclusion, the results show that the lead compounds are effective in decreasing
LC3B levels in the presence of bafilomycin.
Fig. 3. Lead compounds decrease bafilomycin-induced LC3B levels. (A) MIA PaCa-2 cells treated with compounds
6i and 6r (1 µM and 10 µM each) show a decrease in the bafilomycin-induced LC3B levels after 24h. (B) KYSE410
treated withchloroquine (1 and 10 µM) show increase in the LC3B level compared to compound 6r (1 µM and 10
µM) that shows a decrease in the LC3B level. (C) KYSE410 cells treated with compound 6r (1 µM, 10 µM and 50
µM) show a concentration-dependent decrease in the bafilomycin-induced LC3B levels after 24h. (D) KYSE410
cells pre-treated with 10 µM MG132 for 1h shows no significant decrease in the LC3B levels upon treatment with
compound 6r (10 µM, 24 h).
We extended our studies to test the effect of the structural modifications on the autophagy-
related mechanism. We synthesized analogs 11 and 13 that contain the N,N-diethylpentane-1,4-

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diamine fragment, which is an important moiety in chloroquine-mediated inhibition of
autophagy. As expected, we observed an increase in LC3B levels compared to DMSO treatment.
Fig. 4. Compounds 11 and 13 increase LC3B levels. KYSE410 cell were treated with chloroquine, compounds
11 and 13 for 24h (10 µM and 50 µM). All three compounds induce an increased expression of LC3B compared to
the DMSO control.
3.3. Compound 7f arrests cells in G2/M phase
To better elucidate the mechanism of action of the leads, representative compounds were tested
for their effect on altering cell cycle. Although compounds 6i, 6f, and 6r and chloroquine
showed no significant effect on cell cycle, compound 7f arrested cells in G2/M phase (Fig. 4).

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Fig. 4. Compound 7f arrests cells in G2/M phase. Cells were treated with increasing
concentrations of lead compounds, stained with propidium iodide and analyzed by flow
cytometry.
4.
Conclusions
We synthesized 55 new compounds containing a drug-like 2-oxo-1,2-dihydroquinoline-4-
carboxamide fragment to select novel lead compounds to treat ESCC. All compounds were
initially tested in two ESCC cell lines using MTT and colony formation assays and active
compounds were tested in three additional cell lines. Overall, several compounds showed
potency comparable to cisplatin. Mechanistic studies of the lead compounds confirmed inhibition
of autophagy as the mechanism of action. From the cell cycle analysis, 7f was found to be the
only derivative that can cause cell cycle arrest at the G2/M phase. Our study suggests that
development of new anti-cancer drugs based on 2-oxo-1,2-dihydroquinoline-4-carboxamide
warrants further investigations due to their unique mechanism of action.

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5.
EXPERIMENTAL METHODS
5.3. Chemistry
All commercial reagents and anhydrous solvents were purchased and used without further
purification. Column chromatography was performed using normal phase silica gel on a Biotage
IsoleraTM flash chromatography system. Proton nuclear magnetic resonance (1H NMR)
spectroscopy was recorded on a Bruker Avance 300 NMR spectrometers. LC-MS were
performed on a Shimadzu LC-MS-2020 system. The purity of the final products was confirmed
1
to be > 95% as determined by HPLC/MS and H NMR. Acetonitrile/water solvent mixture was
used as an eluent for the HPLC.
5.3.1. 7-Bromoquinoline-2,4-dicarboxylic acid (1) was prepared as previously reported [31].
5.3.2. 7-Bromoquinoline-4-carboxylic acid (2). Decarboxylation was done by refluxing
compound 1 (1 g) in 10 mL of nitrobenzene overnight. The reaction mixture was evaporated
under reduced pressure and the resulting solid was crystallized from hexane to give brown solid
in 67% yield (570 mg).
5.3.3. General synthetic method for compounds (3a-d). The carboxylic acid (0.8 mmol), HATU
(hexafluorophosphate azabenzotriazole tetramethyl uranium) (455 mg, 1.2 mmol) and
triethylamine (324 mg, 3.2 mmol) were stirred for 30 minutes in 20 mL dichloromethane. The
corresponding amine (1.19 mmol) was added to the reaction mixture and stirred overnight. The
resulting solid was filtered, washed with dichloromethane and crystallized from methanol unless
otherwise noted. Purification was carried out over silica gel chromatography till purity reached >
95%.

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5.3.3.1. 7-Bromoquinolin-4-yl)(piperidin-1-yl)methanone (3a). Purification was carried out
using silica gel chromatography (Dichloromethane:MeOH 9.8:0.2). Brown solid (23 mg, 9%
yield). LC-MS m/z 319 [M + H] +; 1H NMR (300 MHz, Methanol-d4) δ 8.97 (d, J = 4.4 Hz, 1H),
8.36 – 8.28 (m, 1H), 7.89 – 7.75 (m, 2H), 7.53 (d, J = 4.4 Hz, 1H), 3.88 (t, J = 5.2 Hz, 2H), 3.19
(q, J = 5.7, 4.0 Hz, 2H), 1.77 (d, J = 4.4 Hz, 4H), 1.49 (dt, J = 11.3, 5.2 Hz, 2H).
5.3.3.2. 7-Bromo-N-(2-methoxyphenyl)quinoline-4-carboxamide (3b). Buff solid (20 mg, 7%
yield). LC-MS m/z 357 [M + H]+; 1H NMR (300 MHz, DMSO-d6) δ 10.06 (s, 1H), 9.06 (d, J =
4.3 Hz, 1H), 8.35 (s, 1H), 8.19 (d, J = 9.1 Hz, 1H), 7.92 – 7.84 (m, 2H), 7.75 (d, J = 4.4 Hz, 1H),
7.24 (d, J = 7.6 Hz, 1H), 7.13 (d, J = 8.5 Hz, 1H), 7.02 (dd, J = 8.1, 6.5 Hz, 1H), 3.85 (s, 3H).
5.3.3.3. 7-Bromo-N-(4-fluorophenyl)quinoline-4-carboxamide (3c). Yellow solid (41 mg, 15%
1
yield). LC-MS m/z 345 [M + H] +; H NMR (300 MHz, Methanol-d4) δ 9.06 – 8.98 (m, 1H),
8.33 (d, J = 2.2 Hz, 1H), 8.19 (dd, J = 9.0, 1.5 Hz, 1H), 7.88 – 7.71 (m, 4H), 7.22 – 7.10 (m, 2H).
5.3.3.4. 7-Bromo-N-(4-chlorophenyl)quinoline-4-carboxamide (3d). White solid (54 mg, 19%
1
yield). LC-MS m/z 361 [M + H] +; H NMR (300 MHz, Methanol-d4) δ 9.05 – 8.95 (m, 1H),
8.30 (d, J = 2.1 Hz, 1H), 8.17 (dd, J = 9.2, 1.5 Hz, 1H), 7.82 (s, 1H), 7.80 – 7.73 (m, 3H), 7.46 –
7.35 (m, 2H).
5.3.4.
6-Methoxy-1H-pyrrolo[2,3-b]pyridine-2,3-dione (4). Pyridinium chlorochromate (1.6
g, 7.5 mmol) was mixed with silica gel (1.6 g, 70-230 mesh) and transferred to a round-bottom
flask containing dichloroethane (20 mL). To the resulting orange suspension was added a
solution of azaindole (450 mg, 3 mmol) in dichloroethane (10 mL) while stirring at room
temperature. To this, AlCl3 (5 mg, 0.039 mmol) was added, and the reaction mixture was stirred
at 80 °C. Completion of the reaction was monitored by TLC. After completion of the reaction,

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the solid was filtered under suction through a sintered funnel layered with silica gel. The filtrate
was evaporated. Crude product was subjected to the next reaction.
5.3.5. General synthetic method for compounds (5a,5c-f and 14). Synthesis was achieved
according to the reported procedure. [32]
5.1.6. General synthetic method for compounds (6a-y, 12 and 15). Same procedure as for
compounds 3a-d.
5.1.6.1. 7-Bromo-4-(piperidine-1-carbonyl)quinolin-2(1H)-one (6a). Brown solid (48 mg, 18%
yield). LC-MS m/z 335 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ 11.98 (s, 1H), 7.54 (d, J =
1.8 Hz, 1H), 7.46 – 7.24 (m, 2H), 6.46 (s, 1H), 3.17 (d, J = 4.8 Hz, 4H), 1.60 (d, J = 4.7 Hz, 4H),
1.44 (s, 2H).
5.1.6.2. 7-Bromo-N-(2-methoxyphenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6b). White
solid (150 mg, 51% yield). LC-MS m/z 373 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ 12.04
(s, 1H), 10.00 (s, 1H), 7.87 – 7.77 (m, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.56 (s, 1H), 7.47 – 7.37 (m,
1H), 7.29 – 7.17 (m, 1H), 7.11 (d, J = 8.2 Hz, 1H), 7.00 (t, J = 7.7 Hz, 1H), 6.69 (s, 1H), 3.84 (s,
3H).
5.1.6.3. 7-Bromo-N-(4-methoxyphenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6c). White
solid (30% yield). LC-MS m/z 373 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ 12.07 (s, 1H),
10.61 (s, 1H), 7.68 (dd, J = 13.5, 8.6 Hz, 3H), 7.56 (s, 1H), 7.39 (d, J = 8.7 Hz, 1H), 6.95 (d, J =
8.5 Hz, 2H), 6.74 (s, 1H), 3.75 (s, 3H).
5.1.6.4. 7-Bromo-N-(4-chlorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6d). Buff
solid (88 mg, 33% yield). LC-MS m/z 377 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ 12.10
(s, 1H), 10.88 (s, 1H), 7.78 (d, J = 8.5 Hz, 2H), 7.69 (d, J = 8.8 Hz, 1H), 7.59 – 7.55 (m, 1H),
7.45 (d, J = 8.2 Hz, 2H), 7.39 (d, J = 9.3 Hz, 1H), 6.79 (s, 1H).

ACCEPTED MANUSCRIPT
5.1.6.5. 7-Bromo-N-(2,4-difluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
(6e).
Yellow solid (66 mg, 22% yield). LC-MS m/z 379 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
7.86 – 7.66 (m, 2H), 7.57 (d, J = 1.9 Hz, 1H), 7.40 (ddd, J = 11.7, 8.9, 2.3 Hz, 2H), 7.17 (dd, J =
9.8, 7.8 Hz, 1H), 6.78 (s, 1H).
5.1.6.6. 7-Bromo-2-oxo-N-(pyridin-2-yl)-1,2-dihydroquinoline-4-carboxamide (6f). Yellow
1
solid (49 mg, 18% yield). LC-MS m/z 344 [M + H] +; H NMR (300 MHz, DMSO-d6) δ 12.05
(s, 1H), 11.27 (s, 1H), 8.39 (d, J = 4.5 Hz, 1H), 8.22 (d, J = 7.9 Hz, 1H), 7.89 (t, J = 8.1 Hz, 1H),
7.69 – 7.48 (m, 2H), 7.43 – 7.33 (m, 1H), 7.22 (t, J = 6.1 Hz, 1H), 6.73 (s, 1H).
5.1.6.7. 7-Bromo-N-(4-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6g). White
solid (86 mg, 30% yield). LC-MS m/z 361 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ 12.08
(s, 1H), 10.81 (s, 1H), 7.86 – 7.64 (m, 3H), 7.57 (d, J = 2.0 Hz, 1H), 7.39 (dd, J = 8.8, 2.0 Hz,
1H), 7.23 (t, J = 8.8 Hz, 2H), 6.78 (s, 1H).
5.1.6.8. 7-Bromo-N-(2,5-difluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
(6h).
Yellow solid (42 mg, 14% yield). LC-MS m/z 379 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
10.76 (s, 1H), 7.84 (s, 1H), 7.70 (d, J = 8.7 Hz, 1H), 7.57 (d, J = 1.8 Hz, 1H), 7.41 (d, J = 8.9 Hz,
2H), 7.15 (s, 1H), 6.78 (s, 1H); 13C NMR (101 MHz, DMSO) δ 164.91, 161.46, 158.06 (d, J
=240.4 Hz, 2.0 Hz), 151.25 (d, J = 245.4 Hz, 2.0 Hz), 144.99, 140.77, 128.09, 126.65 (dd, J =
15.1 Hz, 12.1 Hz), 125.61, 124.54, 121.59, 118.44, 117.26 (dd, J = 23.2 Hz, 9.1 Hz), 115.59,
113.25 (dd, J = 24.2 Hz, 8.1 Hz), 112.48 (d, J = 27.3 Hz).
5.1.6.9. 7-Bromo-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6i). White
solid (111 mg, 39% yield). LC-MS m/z 361 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ 12.15
– 12.07 (m, 1H), 10.95 (s, 1H), 7.71 (t, J = 10.2 Hz, 2H), 7.61 – 7.35 (m, 4H), 7.00 (t, J = 8.2 Hz,
1H), 6.80 (s, 1H); 13C NMR (101 MHz, DMSO) δ 164.55, 162.52 (d, J= 242.2 Hz), 161.51,

ACCEPTED MANUSCRIPT
145.35, 140.82, 140.60 (d, J = 11.1 Hz), 130.96 (d, J = 9.1 Hz), 128.21, 125.65, 124.59, 121.36,
118.45, 116.28 (d, J= 2.0 Hz), 115.46, 111.28 (d, J = 21.2 Hz), 107.33 (d, J = 26.3 Hz).
5.1.6.10. 7-Bromo-N-(2,5-dimethoxyphenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
(6j).
White solid (80 mg, 25% yield). LC-MS m/z 403 [M + H] +; 1H NMR (400 MHz, DMSO-d6) δ
12.03 (s, 1H), 9.96 (s, 1H), 7.71 (d, J = 8.6 Hz, 1H), 7.57 (dd, J = 6.8, 2.5 Hz, 2H), 7.41 (dd, J =
8.7, 2.0 Hz, 1H), 7.03 (d, J = 9.0 Hz, 1H), 6.79 (dd, J = 8.9, 3.1 Hz, 1H), 6.69 (s, 1H), 3.76 (s,
6H).
5.1.6.11. 7-Bromo-2-oxo-N-(4-(trifluoromethoxy)phenyl)-1,2-dihydroquinoline-4-carboxamide
(6k). White solid (68 mg, 20% yield). LC-MS m/z 427 [M + H] +; 1H NMR (300 MHz, DMSO-
d6) δ 12.12 (s, 1H), 10.96 (s, 1H), 7.92 – 7.82 (m, 2H), 7.70 (d, J = 8.6 Hz, 1H), 7.57 (d, J = 2.0
Hz, 1H), 7.40 (td, J = 6.3, 3.2 Hz, 3H), 6.81 (s, 1H).
5.1.6.12. 7-Bromo-2-oxo-N-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-4-carboxamide
(6l). White solid (98 mg, 30% yield). LC-MS m/z 411 [M + H] +; 1H NMR (300 MHz, DMSO-
d6) δ 12.10 (s, 1H), 11.11 (s, 1H), 7.97 (d, J = 8.4 Hz, 2H), 7.77 (d, J = 8.4 Hz, 2H), 7.69 (d, J =
8.6 Hz, 1H), 7.57 (s, 1H), 7.40 (dd, J = 8.5, 1.9 Hz, 1H), 6.84 (s, 1H).
5.1.6.13. 7-Bromo-N-(3-methoxyphenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
(6m).
White solid (77 mg, 26% yield). LC-MS m/z 373 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
12.08 (s, 1H), 10.72 (s, 1H), 7.69 (d, J = 8.6 Hz, 1H), 7.57 (s, 1H), 7.47 – 7.35 (m, 2H), 7.29 (d, J
13
= 5.8 Hz, 2H), 6.75 (d, J = 7.8 Hz, 2H), 3.76 (s, 3H); C NMR (101 MHz, DMSO) δ 164.32,
161.56, 159.99, 145.69, 140.82, 140.10, 130.10, 128.23, 125.63, 124.52, 121.14, 118.43, 115.59,
112.74, 110.27, 106.30, 55.55
5.1.6.14. 7-Bromo-2-oxo-N-(3-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-4-carboxamide
(6n). White solid (59 mg, 18% yield).LC-MS m/z 411 [M + H] +; 1H NMR (300 MHz, DMSO-

ACCEPTED MANUSCRIPT
d6) δ 12.13 (s, 1H), 11.08 (s, 1H), 8.26 (s, 1H), 7.94 (d, J = 8.1 Hz, 1H), 7.73 (d, J = 8.7 Hz, 1H),
7.68 – 7.60 (m, 1H), 7.60 – 7.48 (m, 2H), 7.39 (dd, J = 8.6, 2.0 Hz, 1H), 6.86 (s, 1H).
5.1.6.15. 7-Bromo-N-(4-(tert-butyl)phenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
(6o).
White solid (85 mg, 27% yield). LC-MS m/z 399 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
12.09 (s, 1H), 10.69 (s, 1H), 7.67 (t, J = 8.5 Hz, 3H), 7.56 (d, J = 2.0 Hz, 1H), 7.40 (d, J = 8.5
Hz, 3H), 6.74 (s, 1H), 1.28 (s, 9H).
5.1.6.16. 7-Bromo-2-oxo-N-(3,4,5-trifluorophenyl)-1,2-dihydroquinoline-4-carboxamide (6p).
White solid (170 mg, 54% yield). LC-MS m/z 397 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
12.15 (s, 1H), 11.11 (s, 1H), 7.77 – 7.60 (m, 3H), 7.57 (d, J = 1.8 Hz, 1H), 7.48 – 7.31 (m, 1H),
6.83 (s, 1H).
5.1.6.17 7-Bromo-N-(3-chlorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6q). White
solid (159 mg, 53% yield).LC-MS m/z 377 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ 12.12
(s, 1H), 10.93 (d, J = 6.1 Hz, 1H), 7.96 (s, 1H), 7.78 – 7.52 (m, 3H), 7.40 (dd, J = 8.1, 4.2 Hz,
2H), 7.23 (d, J = 7.6 Hz, 1H), 6.81 (d, J = 4.0 Hz, 1H).
5.1.6.18. 7-Bromo-N-(3,5-difluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6r)
.
White solid (168 mg, 56% yield). LC-MS m/z 379 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
12.15 (s, 1H), 11.12 (s, 1H), 7.70 (d, J = 8.6 Hz, 1H), 7.57 (d, J = 2.0 Hz, 1H), 7.52 – 7.44 (m,
2H), 7.40 (dd, J = 8.7, 2.0 Hz, 1H), 7.06 (tt, J = 9.5, 2.6 Hz, 1H), 6.83 (d, J = 1.5 Hz, 1H).
5.1.6.19. 7-Bromo-2-oxo-N-(3-(trifluoromethoxy)phenyl)-1,2-dihydroquinoline-4-carboxamide
1
(6s). White solid (59 mg, 26% yield). LC-MS m/z 427 [M + H] +; H NMR (300 MHz, DMSO-
d6) δ 12.14 (s, 1H), 11.05 (s, 1H), 7.93 (s, 1H), 7.83 – 6.98 (m, 6H), 6.84 (s, 1H).
5.1.6.20. 7-Fluoro-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6t). White
1
solid (121 mg, 51% yield). LC-MS m/z 301 [M+H]+; H NMR (300 MHz, DMSO-d6) δ 12.15

ACCEPTED MANUSCRIPT
(s, 1H), 10.97 (s, 1H), 7.89 – 7.61 (m, 2H), 7.55 – 7.31 (m, 2H), 7.13 (ddt, J = 13.1, 9.2, 4.5 Hz,
2H), 7.00 (t, J = 8.3 Hz, 1H), 6.72 (s, 1H).
5.1.6.21. N-(3,5-Difluorophenyl)-7-fluoro-2-oxo-1,2-dihydroquinoline-4-carboxamide (6u).
1
White solid (133 mg, 53% yield). LC-MS m/z 319 [M+H]+; H NMR (300 MHz, DMSO-d6) δ
12.17 (s, 1H), 11.12 (s, 1H), 7.81 (dd, J = 8.9, 6.0 Hz, 1H), 7.48 (dd, J = 9.3, 2.3 Hz, 2H), 7.21 –
6.95 (m, 3H), 6.75 (s, 1H).
5.1.6.22. 7-Chloro-N-(3,5-difluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (6v).
1
Brown solid (122 mg, 46% yield). LC-MS m/z 335 [M+H]+, 376 [M+H+ MeCN]+; H NMR
(300 MHz, DMSO-d6) δ 12.16 (s, 1H), 11.12 (s, 1H), 7.78 (d, J = 8.7 Hz, 1H), 7.48 (d, J = 8.6
Hz, 2H), 7.42 (d, J = 2.1 Hz, 1H), 7.28 (dd, J = 8.7, 2.1 Hz, 1H), 7.06 (t, J = 9.3 Hz, 1H), 6.82 (s,
1H).
5.1.6.23. N-(3,5-Difluorophenyl)-2-oxo-7-(trifluoromethyl)-1,2-dihydroquinoline-4-carboxamide
1
(6w). White solid (128 mg, 44% yield). LC-MS m/z 369 [M+H]+, 410 [M+H+ MeCN]+; H
NMR (300 MHz, DMSO-d6) δ 12.34 (s, 1H), 11.17 (s, 1H), 7.99 (d, J = 8.4 Hz, 1H), 7.71 (s,
1H), 7.52 (dd, J = 15.1, 8.6 Hz, 2H), 7.07 (t, J = 9.3 Hz, 1H), 6.99 (s, 1H), 6.56 (s, 1H).
5.1.6.24. N-(3,5-Difluorophenyl)-7-methoxy-2-oxo-1,2-dihydroquinoline-4-carboxamide (6x).
1
Brown solid (126 mg, 48% yield). LC-MS m/z 372 [M + H+ MeCN] +; H NMR (300 MHz,
DMSO-d6) δ 11.93 (s, 1H), 11.06 (s, 1H), 7.65 (d, J = 8.8 Hz, 1H), 7.49 (d, J = 8.8 Hz, 2H), 7.03
(d, J = 9.3 Hz, 1H), 6.86 (dd, J = 12.1, 3.2 Hz, 2H), 6.57 (s, 1H), 3.83 (s, 3H).
5.1.6.25.
N-(3,5-Difluorophenyl)-7-methoxy-2-oxo-1,2-dihydro-1,8-naphthyridine-4-
1
carboxamide (6y). White solid (113 mg, 43% yield). LC-MS m/z 373 [M + H+ MeCN] +; H
NMR (300 MHz, DMSO-d6) δ 12.39 (s, 1H), 11.07 (s, 1H), 8.09 (d, J = 8.8 Hz, 1H), 7.49 (d, J =
9.0 Hz, 2H), 7.05 (s, 1H), 6.83 – 6.60 (m, 2H), 3.95 (s, 3H).

ACCEPTED MANUSCRIPT
5.1.6.26. N-(3,5-Difluorophenyl)-8-methoxy-2-oxo-1,2-dihydroquinoline-4-carboxamide (15).
White solid (102 mg, 39% yield). LC-MS m/z 372 [M + H+ MeCN] +; 1H NMR (300 MHz,
DMSO-d6) δ 11.21 (s, 1H), 11.13 (s, 1H), 7.49 (d, J = 8.8 Hz, 2H), 7.36 – 7.11 (m, 3H), 7.05 (t,
J = 9.2 Hz, 1H), 6.78 (s, 1H), 3.93 (s, 3H).
5.1.7. N-(3,5-Difluorophenyl)-7-hydroxy-2-oxo-1,2-dihydroquinoline-4-carboxamide (6z). To a
solution of 6x (10 mg, 0.03 mmol) in dichloromethane, BBr3 (1M in dichloromethane) (3 equiv)
was added at 0 ºC and stirred at that temperature for 4h. After completion of the reaction as
monitored by TLC analysis, work up was done using dichloromethane and water. The crude
product was purified by column chromatography. White solid (5 mg, 52% yield). LC-MS m/z
358 [M + H+ MeCN] +; 1H NMR (300 MHz, DMSO-d6) δ 11.83 (s, 1H), 11.03 (s, 1H), 10.34 (s,
1H), 7.51 (dd, J = 16.0, 8.8 Hz, 3H), 7.04 (t, J = 9.4 Hz, 1H), 6.77 (s, 1H), 6.69 (d, J = 8.0 Hz,
1H), 6.47 (s, 1H).
5.1.8. N-(3,5-Difluorophenyl)-8-hydroxy-2-oxo-1,2-dihydroquinoline-4-carboxamide (16). Same
procedure as compound 6z. White solid (4 mg, 41% yield). LC-MS m/z 358 [M + H+ MeCN] +;
1H NMR (300 MHz, DMSO-d6) δ 11.09 (s, 1H), 10.89 (s, 1H), 10.50 (s, 1H), 7.48 (d, J = 8.7
Hz, 2H), 7.15 (d, J = 6.4 Hz, 1H), 7.10 – 6.96 (m, 3H), 6.73 (s, 1H).
5.1.9. General synthetic method for compounds (7a-r): To a suspension of compound 6i (0.76
mmol) in 3 mL dioxane:water (4:1), the boronic acid reagent (0.76 mmol) and Cs2CO3 (270 mg,
0.83 mmol) was added. The mixture was purged using nitrogen gas then PdCl2(dppf) (10 mg,
0.014 mmol) was added under inert condition. The mixture was heated to 100 °C for 24 h under
argon and then cooled to room temperature. Filtration using celite, then water (30 mL) was
added and the resulting mixture was extracted with EtOAc. The organic phase was dried over

ACCEPTED MANUSCRIPT
MgSO4, filtered, and dried under vacuum. The crude product was purified by flash
chromatography (Dichloromethane:MeOH 9.8:0.2) to afford the compounds described below.
5.1.9.1. N-(3-Fluorophenyl)-2-oxo-7-phenyl-1,2-dihydroquinoline-4-carboxamide (7a). Brown
solid (77 mg, 28% yield). LC-MS m/z 359 [M + H]+ ; 1H NMR (300 MHz, DMSO-d6) δ 7.88 –
7.75 (m, 2H), 7.70 (d, J = 8.4 Hz, 3H), 7.64 (s, 1H), 7.60 – 7.38 (m, 5H), 7.02 (s, 1H), 6.76 (s,
1H).
5.1.9.2. 7-(4-(Tert-butyl)phenyl)-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
(7b).
Brown solid (50 mg, 16 % yield). LC-MS m/z 415 [M + H]+; 1H NMR (300 MHz, DMSO-
d6) δ 12.11 (s, 1H), 10.98 (s, 1H), 7.96 (s, 1H), 7.81 (d, J = 9.3 Hz, 1H), 7.72 (d, J = 7.9 Hz, 2H),
7.62 (d, J = 5.5 Hz, 1H), 7.54 (d, J = 7.7 Hz, 3H), 7.35 (d, J = 7.7 Hz, 2H), 7.05 – 6.96 (m, 1H),
6.75 (s, 1H), 1.27 (s, 9H).
5.1.9.3. N-(3-Fluorophenyl)-7-(4-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (7c).
Brown solid (132 mg, 46% yield). LC-MS m/z 377 [M + H] +; 1H NMR (300 MHz, DMSO-d6)
δ 12.11 (s, 1H), 10.98 (s, 1H), 7.89 – 7.68 (m, 4H), 7.62 – 7.30 (m, 6H), 7.10 – 6.91 (m, 1H),
6.76 (d, J = 1.6 Hz, 1H).
5.1.9.4. 7-(4-Acetylphenyl)-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (7d).
Brown solid (159 mg, 52% yield). LC-MS m/z 401 [M + H] +; 1H NMR (300 MHz, DMSO-d6)
δ 11.00 (s, 1H), 8.11 (d, J = 8.7 Hz, 2H), 7.95 – 7.26 (m, 8H), 7.02 (s, 1H), 6.80 (s, 1H), 2.64 (s,
3H).
5.1.9.5.
N-(3-Fuorophenyl)-7-(4-methoxyphenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
(7e)
.
Brown solid (98 mg, 33% yield). LC-MS m/z 389 [M + H] +; 1H NMR (300 MHz, DMSO-
d6) δ 12.06 (s, 1H), 10.97 (s, 1H), 7.95 – 7.26 (m, 8H), 7.20 – 6.87 (m, 3H), 6.72 (s, 1H), 3.82 (s,
13
3H); C NMR (101 MHz, DMSO) δ 165.02, 163.76 (d, J = 242.2 Hz), 161.84, 160.13, 145.75,

ACCEPTED MANUSCRIPT
142.75, 140.75 (d, J = 11.1 Hz), 140.38, 131.63, 131.03, 130.94, 128.51 (2C), 126.78, 121.17,
120.18, 116.29 (d, J= 2.0 Hz), 115.12 (2C), 112.98, 111.21 (d, J = 21.2 Hz), 107.31 (d, J = 26.3
Hz), 55.75.
5.1.9.6. N-(3-Fluorophenyl)-2-oxo-7-(thiophen-3-yl)-1,2-dihydroquinoline-4-carboxamide (7f).
Brown solid (45 mg, 16% yield). LC-MS m/z 365 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
12.04 (s, 1H), 10.97 (s, 1H), 7.97 (t, J = 1.9 Hz, 1H), 7.78 (s, 1H), 7.73 (dd, J = 6.0, 3.7 Hz, 2H),
7.65 (s, 1H), 7.60 (dd, J = 8.6, 1.8 Hz, 1H), 7.55 – 7.39 (m, 3H), 7.09 – 6.92 (m, 1H), 6.81 – 6.61
(m, 1H); 13C NMR (101 MHz, DMSO) δ 164.97, 162.55 (d, J = 242.4 Hz), 161.79, 145.69,
140.71, 140.70 (d, J = 11.1 Hz), 140.40, 137.90, 130.99 (d, J = 10.0 Hz), 128.31, 126.81, 126.42,
123.37, 121.15, 120.31, 116.28 (d, J = 2.0 Hz), 115.36, 112.89, 111.22 (d, J = 21.2 Hz), 107.32
(d, J = 26.3 Hz).
5.1.9.7. N,7-bis(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (7g). Brown solid
1
(35 mg, 12% yield). LC-MS m/z 377 [M + H] +; H NMR (300 MHz, DMSO-d6) δ 11.01 (s,
1H), 7.80 (dd, J = 21.5, 9.9 Hz, 2H), 7.66 – 7.35 (m, 7H), 7.29 (dd, J = 9.4, 6.7 Hz, 1H), 7.01 (t, J
= 8.3 Hz, 1H), 6.78 (s, 1H).
5.1.9.8. 7-(2-Fluorophenyl)-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (7h).
White solid (60 mg, 21%yield). LC-MS m/z 377 [M + H] +; 1H NMR (300 MHz, DMSO-d6) δ
12.14 (s, 1H), 10.99 (s, 1H), 7.88 – 7.71 (m, 2H), 7.64 – 7.30 (m, 8H), 7.01 (ddd, J = 10.4, 8.3,
2.1 Hz, 1H), 6.80 (d, J = 1.5 Hz, 1H).
5.1.9.9. N-(3-Fluorophenyl)-2-oxo-7-(p-tolyl)-1,2-dihydroquinoline-4-carboxamide (7i). White
solid (59 mg, 21%yield). LC-MS m/z 373 [M + H]+; 1H NMR (300 MHz, DMSO-d6) δ 12.07 (s,
1H), 10.97 (s, 1H), 7.87 – 7.69 (m, 2H), 7.66 – 7.24 (m, 8H), 7.00 (t, J = 8.4 Hz, 1H), 6.74 (s,
1H), 2.37 (s, 3H).

ACCEPTED MANUSCRIPT
5.1.9.10. 7-Cyclopropyl-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide (7j).
1
White solid (50 mg, 20% yield), LC-MS m/z 324 [M+H]+; H NMR (300 MHz, DMSO-d6) δ
12.05 (s, 1H), 10.95 (s, 1H), 7.79 – 7.69 (m, 2H), 7.58 (t, J = 7.9 Hz, 1H), 7.48 (t, J = 7.1 Hz,
1H), 7.41 (t, J = 8.8 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.07 – 6.92 (m, 1H), 6.74 (s, 1H), 4.13 (q,
J = 5.2 Hz, 1H), 3.17 (d, J = 5.2 Hz, 4H).
5.1.9.11. N-(3-Fluorophenyl)-2-oxo-7-(pyridin-4-yl)-1,2-dihydroquinoline-4-carboxamide (7k).
1
Pale yellow solid (137 mg, 50% yield). LC-MS m/z 360 [M+H]+; 401 [M+H+CH3CN]+. H
NMR (300 MHz, DMSO-d6) δ 12.18 (s, 1H), 11.00 (s, 1H), 8.72 (s, 2H), 7.89 (d, J = 8.4 Hz,
1H), 7.81 – 7.59 (m, 5H), 7.54 – 7.32 (m, 2H), 7.01 (t, J = 7.9 Hz, 1H), 6.83 (s, 1H).
5.1.9.12.
7-(4-Aminophenyl)-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-carboxamide
1
(7l). Yellow solid (96 mg, 34% yield). LC-MS m/z 374 [M+H]+. H NMR (300 MHz, DMSO-
d6) δ 11.95 (s, 1H), 10.94 (s, 1H), 7.73 (dd, J = 15.3, 10.0 Hz, 2H), 7.57 – 7.34 (m, 6H), 7.00 (t, J
= 8.3 Hz, 1H), 6.73 – 6.58 (m, 3H), 5.45 (s, 2H).
5.1.9.13.
7-(3,6-Dihydro-2H-pyran-4-yl)-N-(3-fluorophenyl)-2-oxo-1,2-dihydroquinoline-4-
carboxamide (7m). Yellow solid (45 mg, 16% yield). LC-MS m/z 365 [M+H]+; 406
1
[M+H+CH3CN]+. H NMR (300 MHz, DMSO-d6) δ 12.01 (s, 1H), 10.95 (s, 1H), 7.72 (dd, J =
14.2, 10.4 Hz, 2H), 7.55 – 7.36 (m, 3H), 7.07 – 6.94 (m, 1H), 6.70 (s, 1H), 6.57 (s, 1H), 6.42 (s,
1H), 4.27 (d, J = 2.9 Hz, 2H), 3.85 (t, J = 5.4 Hz, 2H), 2.47 (m, 2H).
5.1.9.14.
N-(3-Fluorophenyl)-7-(6-fluoropyridin-3-yl)-2-oxo-1,2-dihydroquinoline-4-
carboxamide (7n). White solid (160 mg, 56% yield). LC-MS m/z 378 [M+H]+, 419
1
[M+H+CH3CN]+; H NMR (300 MHz, DMSO-d6) δ 12.15 (s, 1H), 10.99 (s, 1H), 8.58 (d, J =
2.5 Hz, 1H), 8.31 (td, J = 8.0, 2.6 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.76 (d, J = 11.3 Hz, 1H),