Design, synthesis and biological evaluation of novel 4-alkynyl-quinoline derivatives as PI3K/mTOR dual inhibitors

Article abstract of DOI:10.1016/j.ejmech.2015.05.025

Abstract A novel series of 4-alkynyl-quinoline derivatives were designed, synthesized and biologically evaluated for their PI3Kα inhibitory activities and anti-proliferative effects against two cancer cell lines PC-3 and HCT-116. Most of them showed potent PI3Kα inhibitory activities with IC₅₀ values at low nanomolar level and good to excellent anti-proliferative effects against both cell lines. Among them, compound 15d, the most potent one, was selected for further biological evaluation. As a result, 15d displayed strong inhibitory activity against other class I PI3K isoforms (PI3Kβ, PI3Kγ and PI3Kδ) and mTOR with an acceptable kinase selectivity profile. Moreover, the western blot assay indicated that the phosphorylation of Akt, another downstream effector of PI3K, can be remarkably suppressed by 15d at cellular level. All these experimental results suggested that 15d is a potent PI3K/mTOR dual inhibitor and could serve as a promising lead compound for the development of anticancer agents.

Full text of DOI:10.1016/j.ejmech.2015.05.025

Accepted Manuscript
Design, synthesis and biological evaluation of novel 4-alkynyl-quinoline derivatives as
PI3K/mTOR dual inhibitors
Xiaoqing Lv, Huazhou Ying, Xiaodong Ma, Ni Qiu, Peng Wu, Bo Yang, Yongzhou Hu
PII:
S0223-5234(15)30052-0
10.1016/j.ejmech.2015.05.025
EJMECH 7904
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 31 October 2014
Revised Date: 9 May 2015
Accepted Date: 15 May 2015
Please cite this article as: X. Lv, H. Ying, X. Ma, N. Qiu, P. Wu, B. Yang, Y. Hu, Design, synthesis and
biological evaluation of novel 4-alkynyl-quinoline derivatives as PI3K/mTOR dual inhibitors, European
Journal of Medicinal Chemistry (2015), doi: 10.1016/j.ejmech.2015.05.025.
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ACCEPTED MANUSCRIPT
Graphic Abstract
Design, synthesis and biological evaluation of novel 4-alkynyl-quinoline derivatives as
PI3K/mTOR dual inhibitors
Xiaoqing Lv, Huazhou Ying, Xiaodong Ma, Ni Qiu, Peng Wu, Bo Yang, Yongzhou Hu

ACCEPTED MANUSCRIPT
Design, synthesis and biological evaluation of novel 4-alkynyl-quinoline
derivatives as PI3K/mTOR dual inhibitors
Xiaoqing Lv, Huazhou Ying, Xiaodong Ma, Ni Qiu, Peng Wu, Bo Yang, Yongzhou Hu*
Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical
Sciences, Zhejiang University, Hangzhou 310058, China
Abstract: A novel series of 4-alkynyl-quinoline derivatives were designed,
synthesized and biologically evaluated for their PI3Kα inhibitory activities and
anti-proliferative effects against two cancer cell lines PC-3 and HCT-116. Most of
them showed potent PI3Kα inhibitory activities with IC values at low nanomolar
5
0
level and good to excellent anti-proliferative effects against both cell lines. Among
them, compound 15d, the most potent one, was selected for further biological
evaluation. As a result, 15d displayed strong inhibitory activity against other class I
PI3K isoforms (PI3Kβ, PI3Kγ and PI3Kδ) and mTOR with an acceptable kinase
selectivity profile. Moreover, the western blot assay indicated that the
phosphorylation of Akt, another downstream effector of PI3K, can be remarkably
suppressed by 15d at cellular level. All these experimental results suggested that 15d
is a potent PI3K/mTOR dual inhibitor and could serve as a promising lead compound
for the development of anticancer agents.
Keyword: PI3K, mTOR, Cancer, Inhibitor, 4-alkynyl-quinoline
*
Corresponding author.
E-mail address: huyz@zju.edu.cn (Y. Hu).
Tel./fax: +86 571 8820 8460.
1

ACCEPTED MANUSCRIPT
1
. Introduction
The phosphoinositide 3-kinase (PI3K)/protein kinase B (PKB, also known as
Akt)/mammalian target of rapamycin (mTOR) pathway is a key signal transduction
system involved in the regulation of cell cycle progression, cell growth, survival and
migration, and intracellular vesicular transport [1-3]. Accumulating studies have
demonstrated that PI3K/Akt/mTOR signaling is frequently dysregulated in human
cancers, resulting in constitutive activation of this signaling network [4-8]. Therefore,
a sizeable effort has been devoted to development of cancer therapies targeting key
kinase PI3K and/or downstream effectors such as Akt and mTOR [9-11]. In particular,
dual inhibition of PI3K and mTOR has been considered as a more effective strategy
for cancer therapy, since it can directly target the most commonly mutated
kinase-PI3K [4-8] in this pathway as well as suppress PI3K-independent activation of
mTOR. Meanwhile, dual inhibition of PI3K and mTOR avoids multiple
mTOR-related negative feedback loops that a selective mTOR inhibitor usually fails
to repress [12-15]. To date, a number of PI3K/mTOR dual inhibitors have been
advanced into clinical trials [14, 16], such as GSK2126458 [17], BEZ235 [18, 19],
BGT226 [20, 21], PF-04691502 [22], GDC-0980 [23] and PKI-587 [24], further
suggesting that simultaneous inhibition of PI3K and mTOR has great potential for the
treatment of cancer (Fig. 1).
<
Insert Fig. 1>
In our previous work, a series of quinoxaline derivatives was identified to be PI3K
inhibitors with favorable in vitro activities [25, 26]. However, the moderate in vivo
efficacy of this class of compounds led us to explore the dual targeting strategies for
an improved potency, especially the dual inhibition of PI3K and mTOR. Herein, we
describe our initial efforts in this field to pursue the desired PI3K/mTOR dual
inhibitors through structural modification of GSK2126458. As a PI3K/mTOR dual
inhibitor under clinical trials, GSK2126458 displayed remarkable in vitro and in vivo
potency, as well as excellent oral bioavailability [17]. Due to the availability of the
co-crystal structure of GSK2126458 with PI3Kγ [17], we investigated their binding
2

ACCEPTED MANUSCRIPT
mode seriously and envisioned that introduction of hydrophilic side chain onto C-4
position of the quinoline ring of GSK2126458 to replace pyridazine ring can not only
improve the water solubility but also explore potential interactions with the residues
located nearby ribose pocket (e.g. Lys802 and Ala805) (Fig. 2). In this study, an
alkyne was employed as a linkage bridge between quinoline ring and hydrophilic
group, affording a series of 4-alkynyl-quinoline derivatives (Fig. 2). All of them were
subsequently prepared and evaluated for their in vitro PI3K inhibitory activities and
anti-proliferative effects.
<
Insert Fig. 2>
2
. Results and discussion
2
.1. Chemistry
The synthetic routes for 4-alkynyl-quinolines 15a–n, 19a–e are outlined in Scheme
–3. Treatment of 5-bromo-2-chloro-3-nitropyridine 1 with sodium methoxide gave
1
5
-bromo-2-methyloxy-3-nitropyridine 2. Reduction of 2 with SnCl in ethyl acetate
2
produced the amine 3. Sulfonylation of 3 with benzenesulfonyl chlorides gave
corresponding sulfamides 4a–f, which were then subjected to palladium-catalyzed
Suzuki coupling with bis(pinacolato)diborane to afford boric acid esters 5a–f
(Scheme 1).
Condensation of 4-bromoaniline 6 with diethyl-2-(ethoxymethylene) malonate
followed by intramolecular cyclization provided ethyl quinoline-3-carboxylate 8.
Following hydrolysis of 8, the newly formed 9 was heated to remove the carboxylic
group, leading to the generation of 4-hydroxyquinoline 10. Treatment of 10 with
POCl afforded 4-chloroquinoline 11, which was then treated with KI to give the
3
4
-iodoquinoline 12. Sonogashira coupling of 12 with alkynes 13a–l in the presence of
Pd(PPh ) Cl provided the corresponding intermediates 14a–l. Coupling 14a–k with
3
2
2
5
a via Suzuki reaction yielded corresponding target compounds 15a–k. Similarly,
compound 15l was prepared by reaction of 16 with 5a (Scheme 2).
Deprotection of the Boc-protected piperazidine derivative 14l with TFA followed
by acylation with acetyl chloride or methanesulfonyl chloride yielded 18a and 18b.
3

ACCEPTED MANUSCRIPT
Subsequently, coupling 18a and 18b with 5a afforded target compounds 15m and 15n,
respectively. Compounds 19a–e were obtained via the Suzuki coupling as described
above (Scheme 3).
<
Insert Schemes 1-3>
2
.2. PI3Kα enzymatic and anti-proliferative assays
All synthesized target compounds were evaluated for their PI3Kα inhibitory
activities and anti-proliferative activities against PC-3 and HCT-116 cell lines by
Kinase-Glo Luminescent assay and Sulforhodamine B (SRB) assay, respectively.
BEZ235 was used as the positive control. The experimental results of PI3Kα
enzymatic assay summarized in Table 1, showed that all the tested 2,
4
-difluorophenyl derivatives (15a–n) exhibited potent PI3Kα inhibitory activities with
IC values in the range of 1.63–32.54 nM, better than that of the positive control
5
0
BEZ235. However, compounds 19a–e with mono-fluoro, methyl, trifluoromethyl or
trifluoromethoxy on the phenyl showed moderate to weak inhibitory activities. These
results suggested that the substitution pattern on the phenyl had a significant effect on
inhibitory activity and the 2,4-difluoro substitutent on the phenyl appeared to be
optimal in this series. On the other hand, among the 2,4-difluorophenyl derivatives
(15a–n), compounds (15a, 15d–h, 15m and 15n) with hydroxyl or amide group at the
end of 4-alkynyl linker displayed more than a 4-fold improvement in the enzymatic
activities compared to the compounds 15b and 15i–k. In addition, compound 15c with
N-methylpiperazine fragment and compound 15l with no 4-alkynyl substituent merely
exhibited moderate inhibitory activity among these derivatives.
According to the results of anti-proliferative assay (Table 1), the
4
-alkynyl-quinoline derivatives showed moderate to potent activities and five
compounds (15d, 15g, 15h, 15n and 19b) exhibited submicromolar potencies against
PC-3 or HCT-116 cell line. Among them, it is noteworthy that compounds 15d and
1
5g with favorable values of molecular parameters (such as ClogP and tPSA) not only
showed potent PI3Kα inhibitory potencies but also displayed excellent
anti-proliferative activities. More encouragingly, 4-hydroxylpiperidine derivative 15d,
4

ACCEPTED MANUSCRIPT
the most potent compound in both the PI3K enzymatic and anti-proliferative assays,
showed stronger activity against PC-3 cell line than that of BEZ235. Therefore, this
compound can be used as a promising lead compound for further biological
evaluation.
<
Insert Table 1>
2
.4. Kinase selectivity assay
In order to evaluate the activities of target compounds against other class I PI3Ks
PI3Kβ, PI3Kγ and PI3Kδ) and mTOR, compound 15d was screened for its activities
(
by ADP-Glo Luminescent assay and Lance Ultra assay, respectively. BEZ235 was
used as the positive control. As shown in Table 2, compound 15d displayed
inhibitory activities with IC values ranging from single-digit nanomolar (PI3Kα, β, δ
5
0
and mTOR) to subnanomolar (PI3Kγ), which were more potent than that of BEZ235.
This result suggested that compound 15d was a potent PI3K/mTOR dual inhibitor.
<
Insert Table 2>
Subsequently, 15d was submitted for screening in a panel of 50 kinases provided by
DiscoveRx’s KinomeScan service [27]. The results further confirmed that 15d was a
potent PI3K family and mTOR inhibitor showing moderate-to-good selectivity over
other kinases with the exception of the PI4KIIIβ (PIK4CB) (Table 3).
<
Insert Table 3>
2
.5. Western blot assay
Finally, to determine whether 15d suppressed the activation of Akt, a key node in
the PI3K/Akt/mTOR signaling pathway, it was tested for its suppressive effect on
pAkt(Ser473) level in PC-3 cells. Glyceraldehyde-3-phosphate dehydrogenase
(GAPDH) was used as the internal control. The suppressive effect for 15d was
evaluated at the concentrations of 1 nM, 5 nM, 25 nM and 125 nM. As illustrated in
Fig. 4, at the concentration of 125 nM, 15d exhibited significantly inhibitory effect on
5

ACCEPTED MANUSCRIPT
pAkt(Ser473), which further demonstrated 15d can strongly down-regulate the
PI3K/Akt/mTOR pathway.
<
Insert Fig. 3>
2
.3. Molecular docking study
To examine structure–activity relationships (SARs) in more detail, docking analysis
of 15d bound to PI3Kγ (PDB code 3L08), and 15d, 19a and 19b bound to PI3Kα
PDB code 4JPS) was performed utilizing the Discovery Studio 2.1 software package,
(
respectively. The result of 15d bound to PI3Kγ as shown in Fig. 4, the nitrogen of
sulfonamide in 15d is involved in the hydrogen bonding interaction with Lys833 and
the 4-hydroxy of the piperidine moiety at the end of 4-alkynyl linker forms an
additional hydrogen bond with Lys802 near the ribose pocket. Additionally, a
conserved water molecule is bridged between pyridyl nitrogen and residues Asp841
and Tyr867, and the nitrogen of quinoline makes a critical hydrogen bond with
Val882 in the hinge region, which is consistent with the co-crystal structure of
GSK2126458 bound to PI3Kγ. In addition, in the back pocket, the 2-fluoro and
4
-fluoro on the phenyl moiety of 15d form two hydrogen bonds with the residues
Lys833 and Asn951, respectively. In contrast, the docking result of 15d with PI3Kα
showed the absence of one hydrogen bond between the 4-hydroxy of the piperidine
moiety of 15d and PI3Kα near the ribose pocket. In the other regions, 15d forms five
hydrogen bonds with residues (Lys802, Asp810, Tyr836, Val851 and Asn920) when
bound to PI3Kα. These results were consistent with the molecule’s (15d) potency and
selectivity profiles (subnanomolar PI3Kγ potency and > 4-fold selectivity over
PI3Kα).
The binding modes of 19a, 19b and 15d bound to PI3Kα are generally identical.
The difference lies in the interaction with the back pocket. In detail, 2-fluoro and
4
-fluoro on the phenyl moiety of 15d form two hydrogen bonds with the residues
Lys802 and Asn920 in this region, respectively. However, as for 19a and 19b, only
one hydrogen bond was formed between the 4-fluro or 3-fluro of the phenyl moiety
6

ACCEPTED MANUSCRIPT
and the residue Asn920, thereby accounting for their significant loss in PI3Kα
activities.
<
Insert Fig. 4>
3
. Conclusion
A novel series of 4-alkynyl-quinoline derivatives was evaluated as PI3K/mTOR
dual inhibitors with potent enzymatic and cellular activities, and 15d was identified as
a potentially interesting lead molecule. Most of derivatives exhibited stronger PI3Kα
inhibitory activities than that of BEZ235. Several compounds displayed potent
anti-proliferative activities with IC values less than 1 µM, which were comparable
5
0
to that of BEZ235. Additionally, the most potent compound 15d with an acceptable
kinase selectivity profile significantly inhibited other PI3Ks and mTOR, as well as the
phosphorylation of pAkt(Ser473) at nanomolar level. According to these experimental
results, it can be concluded that derivatization at the C-4 position of the quinoline core
via introducing appropriate substituents to the alkynyl linker is a feasible way to attain
promising inhibitors with high enzymatic and anti-proliferative activities.
4
. Experimental section
4
.1. Chemistry and chemical methods
1
13
1
H NMR and C NMR spectra were recorded on the BRUKER AVII 400 ( H: 400,
1
3
1
13
C: 100 MHz) and AVIII 500 ( H: 500, C: 125 MHz) NMR instruments. Chemical
shifts are given in ppm (δ) relative to TMS as internal standard, coupling constants (J)
are in hertz (Hz), and signals are using the following abbreviations: s, singlet; d,
doublet; dd, doublet of doublets; t, triplet; td, doublet of triplets; q, quartet; m,
multiplet, etc. Mass spectra (MS) were measured on an Esquire-LC-00075
spectrometer. Column chromatography and thin layer chromatography (TLC) were
carried out using silica gel ZCX-3 and GF-254 (Qingdao haiyang chemical Co., Ltd.),
respectively. Reagents and solvents were commercially available without further
purification.
7

ACCEPTED MANUSCRIPT
4
.1.1. 5-Bromo-2-methoxy-3-nitropyridine (2)
To a solution of 5-bromo-2-chloro-3-nitropyridine (1) (10.0 g, 42.37 mmol) in
anhydrous methanol (40 mL) was added sodium methoxide solution (20 mL, 63.56
mmol) slowly at 0 °C. The reaction mixture was then heated to 50 °C and stirred for
1
2 h. After the completion of reaction, the mixture was filtered and the precipitate was
washed with water. The obtained solids were then dried under reduced pressure to
give the title compound (8.12 g, 35.0 mmol, 83% yield) with light yellow. ESI-MS:
+
m/z = 233 [M+H] .
4
.1.2. 5-Bromo-2-methoxypyridin-3-amine (3)
A mixture of 5-bromo-2-methoxy-3-nitropyridine (2) (5.0 g, 21.55 mmol) and tin(II)
chloride dihydrate (24.35 g, 107.75 mmol) was dissolved into ethyl acetate (0.2 L)
and stirred at 50 °C for 3 h under a nitrogen atmosphere. The mixture was dissolved
in EtOAc (0.8 L), washed with 1 N sodium hydroxide (1.2 L) twice, water twice,
dried over magnesium sulfate and concentrated to give the title compound (3.31 g,
1
1
6.39 mmol, 76 % yield) as a brown solid. H NMR (500 MHz, DMSO-d ) δ 7.37 (d,
6
J = 2.0 Hz, 1H, Ar-H), 6.97 (d, J = 2.0 Hz, 1H, Ar-H), 5.28 (s, 2H, NH ), 3.82 (s, 3H,
2
+
OCH ). ESI-MS: m/z = 203 [M+H] .
3
4
.1.3.
General
procedure
A
for
synthesis
of
N-(5-bromo-2-methoxypyridin-3-yl)benzenesulfonamides (4a-f)
To a solution of 5-bromo-2-methoxypyridin-3-amine (3) (1.0 equiv) in anhydrous
pyridine was added benzenesulfonyl chloride (1.0 equiv) at room temperature. The
reaction was then stirred at room temperature for 24 h. The pyridine was removed
under reduced pressure and the residue was purified by silica gel chromatography
(10% ethyl acetate/petroleum ether to 100% ethyl acetate) to give the crude product. It
was further washed with diethyl ether to give the title compound.
4
.1.3.1. N-(5-Bromo-2-methoxypyridin-3-yl)-2,4- difluorobenzenesulfonamide (4a)
This compound was prepared from 5-bromo-2-methoxypyridin-3-amine (3) (3.0 g,
8

ACCEPTED MANUSCRIPT
1
4.85 mmol) and 2,4-difluorobenzenesulfonyl chloride (3.15 g, 14.85 mmol)
according to the general synthesis procedure A to afford the title compound (3.58 g,
1
9
.47 mmol, 64 % yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 10.41 (s,
6
1
H, NH), 8.10 (d, J = 2.5 Hz, 1H, Ar-H), 7.75 (m, 1H, Ar-H), 7.74 (d, J = 2.5 Hz, 1H,
Ar-H), 7.54 (td, J = 8.5, 2.0 Hz, 1H, Ar-H), 7.21 (td, J = 8.5, 2.0 Hz, 1H, Ar-H), 3.60
+
(
s, 3H, OCH ). ESI-MS: m/z = 379 [M+H] .
3
4
.1.3.2. N-(5-Bromo-2-methoxypyridin-3-yl)-4-fluorobenzenesulfonamide (4b)
This compound was prepared from 5-bromo-2-methoxypyridin-3-amine (3) (3.0 g,
4.85 mmol) and 4-fluorobenzenesulfonyl chloride (2.88 g, 14.85 mmol) according to
1
the general synthesis procedure A to afford the title compound (3.15 g, 8.75 mmol,
1
5
8
9% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 10.19 (s, 1H, NH),
6
.08 (d, J = 2.5 Hz, 1H, Ar-H), 7.84 – 7.81 (m, 2H, Ar-H), 7.72 (d, J = 2.5 Hz, 1H,
Ar-H), 7.43 (t, J = 8.5 Hz, 2H, Ar-H), 3.64 (s, 3H, OCH ). ESI-MS: m/z = 361
3
+
[
M+H] .
4
.1.3.3. N-(5-Bromo-2-methoxypyridin-3-yl)-3-fluorobenzenesulfonamide (4c)
This compound was prepared from 5-bromo-2-methoxypyridin-3-amine (3) (3.0 g,
4.85 mmol) and 3-fluorobenzenesulfonyl chloride (2.88 g, 14.85 mmol) according to
1
the general synthesis procedure A to afford the title compound (2.86 g, 7.94 mmol,
1
5
8
7
3% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 10.31 (s, 1H, NH),
6
.09 (d, J = 2.5 Hz, 1H, Ar-H), 7.73 (d, J = 2.5 Hz, 1H, Ar-H), 7.65 (m, 1H, Ar-H),
.59 (m, 2H, Ar-H), 7.55 (m, 1H, Ar-H), 3.64 (s, 3H, OCH ). ESI-MS: m/z = 361
3
+
[
M+H] .
4
.1.3.4. N-(5-Bromo-2-methoxypyridin-3-yl)-4-(trifluoromethyl)benzenesulfonamide
4d)
This compound was prepared from 5-bromo-2-methoxypyridin-3-amine (3) (3.0 g,
4.85 mmol) and 4-(trifluoromethyl)benzene-1-sulfonyl chloride (3.62 g, 14.85 mmol)
(
1
according to the general synthesis procedure A to afford the title compound (3.31 g,
9

ACCEPTED MANUSCRIPT
1
8
1
2
.07 mmol, 54% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 10.42 (s,
6
H, NH), 8.11 (d, J = 2.5 Hz, 1H, Ar-H), 7.97 (q, J = 8.5 Hz, 4H, Ar-H), 7.77 (d, J =
+
.5 Hz, 1H, Ar-H), 3.55 (s, 3H, OCH ). ESI-MS: m/z = 411 [M+H] .
3
4
.1.3.5. N-(5-Bromo-2-methoxypyridin-3-yl)-4-(trifluoromethoxy)benzenesulfonamide
(
4e)
This compound was prepared from 5-bromo-2-methoxypyridin-3-amine (3) (3.0 g,
4.85 mmol) and 4-(trifluoromethoxy)benzene-1-sulfonyl chloride (3.86 g, 14.85
1
mmol) according to the general synthesis procedure A to afford the title compound
1
(
3.55 g, 8.33 mmol, 56% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.28 (s, 1H, NH), 8.11 (d, J = 2.0 Hz, 1H, Ar-H), 7.86 (d, J = 8.5 Hz, 2H, Ar-H),
7
.75 (d, J = 2.0 Hz, 1H, Ar-H), 7.59 (d, J = 8.5 Hz, 2H, Ar-H), 3.57 (s, 3H, OCH3).
+
ESI-MS: m/z = 427 [M+H] .
4
.1.3.6. N-(5-Bromo-2-methoxypyridin-3-yl)-4-methylbenzenesulfonamide (4f)
This compound was prepared from 5-bromo-2-methoxypyridin-3-amine (3) (3.0 g,
4.85 mmol) and 4-methylbenzene-1-sulfonyl chloride (2.82 g, 14.85 mmol)
1
according to the general synthesis procedure A to afford the title compound (3.02 g,
1
8
1
8
.46 mmol, 57% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 10.07 (s,
6
H, NH), 8.03 (d, J = 2.5 Hz, 1H, Ar-H), 7.68 (d, J = 8.0 Hz, 1H, Ar-H), 7.66 (d, J =
.0 Hz, 2H, Ar-H), 7.38 (d, J = 8.0 Hz, 2H, Ar-H), 3.66 (s, 3H, OCH ), 2.37 (s, 3H,
3
+
CH ). ESI-MS: m/z = 357 [M+H] .
3
4
.1.4.
General
procedure
B
for
synthesis
of
N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)benzenesul
fonamides (5a-f)
To a two neck flask with a magnetic stir bar was added bis(pinacolato)diborane (1.0
equiv),
N-(5-bromo-2-methoxypyridin-3-yl)benzenesulfonamide
(1.0
equiv),
Pd(dppf) Cl (0.1 equiv), KOAc (3.0 equiv) and anhydrous dioxane. The mixture was
2
2
deoxygenated by bubbling nitrogen through it for 10 min. The mixture was then
1
0

ACCEPTED MANUSCRIPT
stirred at 100 °C for 3 h. After cooling to room temperature, the mixture was
evaporated under reduced pressure and the residue was dissolved in EtOAc, washed
with water twice and dried over magnesium sulfate. The crude product was purified
by flash chromatography (25% ethyl acetate/petroleum ether) to give the target
compound.
4
2
.1.4.1.
,4-Difluoro-N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-
yl)benzenesulfonamide (5a)
This compound was prepared from bis(pinacolato)diborane (335 mg, 1.32 mmol),
N-(5-bromo-2-methoxypyridin-3-yl)-2,4-difluorobenzenesulfonamide (4a) (499 mg,
1
.32 mmol), Pd(dppf) Cl (97 mg, 0.13 mmol) and KOAc (388 mg, 3.96 mmol)
2 2
according to the general synthesis procedure B to afford the title compound (495 mg,
1
1
1
.16 mmol, 88% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 10.17 (s,
6
H, NH), 8.20 (d, J = 1.5 Hz, 1H, Ar-H), 7.71 (d, J = 1.5 Hz, 1H, Ar-H), 7.69 (m, 1H,
Ar-H), 7.56 (td, J = 8.5, 2.5 Hz, 1H, Ar-H), 7.19 (td, J = 8.5, 2.5 Hz, 1H, Ar-H), 3.62
+
(
s, 3H, OCH ), 1.29 (s, 12H, CH × 4). ESI-MS: m/z = 427 [M+H] .
3
3
4
4
.1.4.2.
-Fluoro-N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)b
enzenesulfonamide (5b)
This compound was prepared from bis(pinacolato)diborane (335 mg, 1.32 mmol),
N-(5-bromo-2-methoxypyridin-3-yl)-4-fluorobenzenesulfonamide (4b) (475 mg, 1.32
mmol), Pd(dppf) Cl (97 mg, 0.13 mmol) and KOAc (388 mg, 3.96 mmol) according
2
2
to the general synthesis procedure B to afford the title compound (413 mg, 1.01 mmol,
1
7
7% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 9.91 (s, 1H, NH), 8.18
6
(d, J = 1.5 Hz, 1H, Ar-H), 7.75 – 7.71 (m, 3H, Ar-H), 7.40 (t, J = 8.5 Hz, 2H, Ar-H),
+
3
.62 (d, J = 4.5 Hz, 3H, OCH ), 1.30 (s, 12H, CH × 4). ESI-MS: m/z = 409 [M+H] .
3
3
4
.1.4.3.
11

ACCEPTED MANUSCRIPT
3
-Fluoro-N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)b
enzenesulfonamide (5c)
This compound was prepared from bis(pinacolato)diborane (335 mg, 1.32 mmol),
N-(5-bromo-2-methoxypyridin-3-yl)-3-fluorobenzenesulfonamide (4c) (475 mg, 1.32
mmol), Pd(dppf) Cl (97 mg, 0.13 mmol) and KOAc (388 mg, 3.96 mmol) according
2
2
to the general synthesis procedure B to afford the title compound (433 mg, 1.06 mmol,
1
8
8
0% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 10.06 (s, 1H, NH),
6
.19 (d, J = 1.5 Hz, 1H, Ar-H), 7.73 (d, J = 1.5 Hz, 1H, Ar-H), 7.64 – 7.60 (m, 1H,
Ar-H), 7.55-7.48 (m, 3H, Ar-H), 3.64( s, 3H, OCH ), 1.30 (s, 12H, CH × 4). ESI-MS:
3
3
+
m/z = 409 [M+H] .
4
.1.4.4.
N-(2-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-4-(trifluor
omethyl)benzenesulfonamide (5d)
This compound was prepared from bis(pinacolato)diborane (335 mg, 1.32 mmol),
N-(5-bromo-2-methoxypyridin-3-yl)-4-(trifluoromethyl)benzenesulfonamide
(4d)
(
541 mg, 1.32 mmol), Pd(dppf) Cl (97 mg, 0.13 mmol) and KOAc (388 mg, 3.96
2
2
mmol) according to the general synthesis procedure B to afford the title compound
1
(
445 mg, 0.97 mmol, 73% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
7
1
0.15 (s, 1H, NH), 8.20 (d, J = 1.5 Hz, 1H, Ar-H), 7.97 (d, J = 8.5 Hz, 2H, Ar-H),
.88 (d, J = 8.5 Hz, 2H, Ar-H), 7.69 (d, J = 1.5 Hz, 1H, Ar-H), 3.55 (s, 3H, OCH3),
+
.30 (s, 12H, CH × 4). ESI-MS: m/z = 459 [M+H] .
3
4
.1.4.5.
N-(2-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-4-(trifluor
omethoxy)benzenesulfonamide (5e)
This compound was prepared from bis(pinacolato)diborane (335 mg, 1.32 mmol),
N-(5-bromo-2-methoxypyridin-3-yl)-4-(trifluoromethoxy)benzenesulfonamide
(4e)
(
562 mg, 1.32 mmol), Pd(dppf) Cl (97 mg, 0.13 mmol) and KOAc (388 mg, 3.96
2
2
mmol) according to the general synthesis procedure B to afford the title compound
1
2

ACCEPTED MANUSCRIPT
1
(
461 mg, 0.97 mmol, 73% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
7
1
0.01 (s, 1H, NH), 8.20 (d, J = 1.5 Hz, 1H, Ar-H), 7.79 (d, J = 8.5 Hz, 2H, Ar-H),
.70 (d, J = 1.5 Hz, 1H, Ar-H), 7.57 (d, J = 8.5 Hz, 2H, Ar-H), 3.57 (s, 3H, OCH3),
+
.30 (s, 12H, CH × 4). ESI-MS: m/z = 475 [M+H] .
3
4
.1.4.6.
N-(2-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-4-methylb
enzenesulfonamide (5f)
This compound was prepared from bis(pinacolato)diborane (335 mg, 1.32 mmol),
N-(5-bromo-2-methoxypyridin-3-yl)-4-methylbenzenesulfonamide (4f) (470 mg, 1.32
mmol), Pd(dppf) Cl (97 mg, 0.13 mmol) and KOAc (388 mg, 3.96 mmol) according
2
2
to the general synthesis procedure B to afford the title compound (423 mg, 1.05 mmol,
1
8
0% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 9.76 (s, 1H, NH), 8.14
6
(d, J = 1.5 Hz, 1H, Ar-H), 7.73 (d, J = 1.5 Hz, 1H, Ar-H), 7.58 (d, J = 8.0 Hz, 2H,
Ar-H), 7.35 (d, J = 8.0 Hz, 2H, Ar-H), 3.64 (s, 3H, OCH ), 2.36 (s, 3H, CH ), 1.30 (s,
3
3
+
1
2H, CH × 4). ESI-MS: m/z = 405 [M+H] .
3
4
.1.5. Ethyl 6-bromo-4-hydroxyquinoline-3-carboxylate (8)
mixture of 4-bromoaniline (10.0 g,
A
58.48
mmol)
and
diethyl-2-(ethoxymethylene)malonate (12.63 g, 58.48 mmol) was stirred at 80 °C for
2
2
h. The mixture was evaporated under reduced pressure to give the diethyl
-((4-bromophenylamino)methylene)malonate (7). This product was then treated with
Ph O (100 mL) and stirred at 250 °C for 6 h. After cooling to 60 °C, petroleum ether
2
(100 mL) was added to the mixture and the suspended solid was filtered, washed with
petroleum ether, EtOAc and dried under reduced pressure to give the title compound
1
(
10.52 g, 35.66 mmol, 61% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
8
1
2.46 (s, 1H, OH), 8.60 (s, 1H, Ar-H), 8.23 (d, J = 2.5 Hz, 1H, Ar-H), 7.88 (dd, J =
.5, 2.5 Hz, 1H, Ar-H), 7.61 (d, J = 8.5 Hz, 1H, Ar-H), 4.23 (q, J = 7.0 Hz, 2H, CH2),
+
.29 (t, J = 7.0 Hz, 3H, CH ). ESI-MS: m/z = 296 [M+H] .
3
1
3

ACCEPTED MANUSCRIPT
4
.1.6. 6-Bromo-4-hydroxyquinoline-3-carboxylic acid (9)
Ethyl 6-bromo-4-hydroxyquinoline-3-carboxylate (8) (6.0 g, 20.34 mmol) and 2.5
N NaOH (50 mL) were charged in a 100 mL round-bottomed flask. The mixture was
stirred at reflux for 1 h. After cooling to room temperature, the pH of the mixture was
adjusted to 5 using 2 N HCl and the resulting solid was filtered and washed with
water. The filter cake was then dried under reduced pressure to afford the title
1
compound (5.15 g, 19.22 mmol, 94% yield) as a white solid. H NMR (500 MHz,
DMSO-d ) δ 12.51 (s, 1H), 8.63 (s, 1H, Ar-), 8.23 (d, J = 2.5 Hz, 1H, Ar-H), 7.87 (dd,
6
J = 8.5, 2.5 Hz, 1H, Ar-H), 7.61 (d, J = 8.5 Hz, 1H, Ar-H). ESI-MS: m/z = 268
+
[
M+H] .
4
.1.7. 6-Bromoquinolin-4-ol (10)
-Bromo-4-hydroxyquinoline-3-carboxylic acid (9) (5.0 g, 18.73 mmol) was added
6
to a 100 mL round-bottomed flask in Ph O (30 mL) and then stirred at 260 °C for 2 h.
2
After cooling to 60 °C, 30 mL petroleum ether was added and the suspended solid
was filtered, washed with petroleum ether, EtOAc and dried under reduced pressure to
give the title compound (3.21 g, 14.39 mmol, 77% yield) as a brown solid. ESI-MS:
+
m/z = 224 [M+H] .
4
.1.8. 6-Bromo-4-chloroquinoline (11)
To a 100 mL round-bottomed flask was added 6-bromoquinolin-4-ol (10) (3.0 g,
1
3.45 mmol), POCl (20 mL) and DMF (0.5 mL). The mixture was stirred at reflux
3
for 6 h. After cooling to room temperature, the reaction mixture was poured into ice
water (100 mL) and stirred for 1 h. Then the pH of the mixture was adjusted to 8
using saturated aqueous NaHCO . The mixture was extracted with EtOAc and the
3
organic phase was dried over sodium sulfate and concentrated in vacuo to give the
1
title compound (2.75 g, 11.36 mmol, 84% yield) as a white solid. H NMR (500 MHz,
DMSO-d ) δ 8.87 (d, J = 4.5 Hz, 1H, Ar-H), 8.32 (d, J = 2.0 Hz, 1H, Ar-H), 8.03 (d, J
6
=
9.0 Hz, 1H, Ar-H), 7.99 (dd, J = 9.0, 2.0 Hz, 1H, Ar-H), 7.82 (d, J = 4.5 Hz, 1H,
+
Ar-H). ESI-MS: m/z = 242 [M+H] .
1
4

ACCEPTED MANUSCRIPT
4
.1.9. 6-Bromo-4-iodoquinoline (12)
To a solution of 6-bromo-4-chloroquinoline (11) (3.50 g, 14.46 mmol) in
anhydrous EtOAc (20 mL) was added HCl-saturated EtOAc (40 mL) and a white
precipitate formed immediately. After stirring for 30 min, the suspension was
concentrated under vacuum to afford 6-bromo-4-chloroquinoline hydrochloride as an
off white solid (3.91 g, 14.14 mmol).
A two-neck flask was charged with 6-bromo-4-chloroquinoline hydrochloride (3.91
g, 14.14mmol), anhydrous potassium iodide (9.76 g, 70.70 mmol) and anhydrous
acetonitrile (100 mL). The resulting slurry was stirred at reflux for 48 h and allowed
to cool to room temperature. Saturated aqueous NaHCO solution (40 mL) was added
3
to the mixture, followed by 20 mL of a 5% sodium sulfite solution. The reaction
mixture was extracted with CH Cl (200 mL × 2). The combined organic extracts
2
2
were dried over magnesium sulfate and concentrated in vacuo to give the crude
product, which was further purified by silica gel column chromatography (25% ethyl
acetate/petroleum ether) to give the title compound (4.42 g, 13.27 mmol, 94% yield)
1
as an off-white solid. H NMR (500 MHz, DMSO-d ) δ 8.51 (d, J = 4.5 Hz, 1H,
6
Ar-H), 8.21 (d, J = 4.5 Hz, 1H, Ar-H), 8.11 (t, J = 1.5 Hz, 1H, Ar-H), 7.97 – 7.91 (m,
+
2
H, Ar-H). ESI-MS: m/z = 334 [M+H] .
4
.1.10. General procedure C for synthesis of aliphatic propargylamines (13b-h and
1
3l)
A solution of 3-bromopropyne (1.0 equiv), amine (1.0 equiv) and K CO (2.0 equiv)
2
3
in THF was stirred at reflux for 6 h. The solvent was removed and the residue was
partitioned between water and CH Cl . The organic layer was dried over magnesium
2
2
sulfate and under reduced pressure. The crude product was further purified by silica
gel column chromatography to give the desired propargylamine.
4
.1.10.1. 1-(Prop-2-ynyl)piperidine (13b)
1
5

ACCEPTED MANUSCRIPT
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
piperidine (360 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol) according to the
2
3
general synthesis procedure C to afford the title compound (344 mg, 2.80 mmol, 66%
+
yield) as a viscous oil. ESI-MS: m/z = 124 [M+H] .
4
.1.10.2. 1-Methyl-4-(prop-2-ynyl)piperazine (13c)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
N-methylpiperazine (424 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol) according
2
3
to the general synthesis procedure C to afford the title compound (359 mg, 2.59 mmol,
1
6
1% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 3.24 (d, J = 2.5 Hz, 2H,
6
CH ), 3.14 (t, J = 2.5 Hz, 1H, acetylenic hydrogen), 2.44 (s, 4H, CH × 2), 2.30 (s, 4H,
2
2
+
CH × 2), 2.15 (s, 3H, CH ). ESI-MS: m/z = 139 [M+H] .
2
3
4
.1.10.3. 1-(Prop-2-ynyl)piperidin-4-ol (13d)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
-hydroxypiperidine (428 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol) according
4
2
3
to general synthesis procedure C to afford the title compound (403 mg, 2.90 mmol,
1
6
8% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 4.53 (d, J = 3.5 Hz, 1H,
6
OH), 3.48 – 3.31 (m, 1H, CH), 3.19 (d, J = 2.0 Hz, 2H, CH ), 3.09 (d, J = 2.0 Hz, 1H,
2
acetylenic hydrogen), 2.69 – 2.59 (m, 2H, CH ), 2.14 (t, J = 9.5 Hz, 2H, CH ), 1.68 (d,
2
2
+
J = 9.5 Hz, 2H, CH ), 1.41 – 1.30 (m, 2H, CH ). ESI-MS: m/z = 140 [M+H] .
2
2
4
.1.10.4. 1-(Prop-2-ynyl)piperidin-3-ol (13e)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
-hydroxypiperidine (428 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol) according
3
2
3
to general synthesis procedure C to afford the title compound (392 mg, 2.82 mmol,
1
6
7% yield) as a white solid. H NMR (500 MHz, DMSO-d ) δ 4.60 (d, J = 5.0 Hz, 1H,
6
OH), 3.43 (m, 1H, CH), 3.21 (d, J = 2.0 Hz, 2H, CH ), 3.10 (d, J = 2.0 Hz, 1H,
2
acetylenic hydrogen), 2.74 (m, 1H, CH ), 2.55 (m, 1H, CH ), 1.98 (m, 1H, CH ), 1.86
2
2
2
(
m, 1H, CH ), 1.74 (m, 1H, CH ), 1.64 – 1.56 (m, 1H, CH ), 1.42 – 1.32 (m, 1H, CH ),
2 2 2 2
1
6

ACCEPTED MANUSCRIPT
+
0
4
2
.99 (m, 1H, CH ). ESI-MS: m/z = 140 [M+H] .
2
.1.10.5. 2-(Methyl(prop-2-ynyl)amino)ethanol (13g)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
-methylaminoethanol (318 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol)
2
3
according to the general synthesis procedure C to afford the title compound (271 mg,
1
2
.40 mmol, 57% yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 4.42 (s, 1H,
6
OH), 3.45 (t, J = 6.5 Hz, 2H, CH ), 3.30 (d, J = 2.5 Hz, 2H, CH ), 3.10 (t, J = 2.5 Hz,
2
2
1
H, acetylenic hydrogen), 2.43 (t, J = 6.5 Hz, 2H, CH ), 2.21 (s, 3H, CH ). ESI-MS:
2 3
+
m/z = 114 [M+H] .
4
.1.10.6. 3-(Methyl(prop-2-ynyl)amino)propane-1,2-diol (13h)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
-methylamino-1,2-propanediol (445 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol)
3
2
3
according to general synthesis procedure C to afford the title compound (314 mg, 2.18
1
mmol, 51% yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 4.48 (s, 1H,
6
OH), 4.41 (d, J = 4.5 Hz, 1H, OH), 3.54 (m, 1H, CH), 3.34 – 3.27 (m, 4H, CH × 2),
2
3
2
.10 (t, J = 2.5 Hz, 1H, acetylenic hydrogen), 2.40 (dd, J = 12.5, 5.0 Hz, 1H, CH2),
+
.28 (dd, J = 12.5, 7.0 Hz, 1H, CH ), 2.23 (s, 3H, CH ). ESI-MS: m/z = 144 [M+H] .
2
3
4
.1.10.7. Tert-butyl 4-(prop-2-ynyl)piperazine-1-carboxylate (13l)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
N-(tert-butoxycarbonyl)piperazine (789 mg, 4.24 mmol) and K CO (1.17 g, 8.48
2
3
mmol) according to the general synthesis procedure C to afford the title compound
1
(
515 mg, 2.30 mmol, 54% yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ
6
3
.33 (m, 4H, CH × 2), 3.29 (d, J = 2.5 Hz, 2H, CH ), 3.18 (t, J = 2.5 Hz, 1H,
2 2
acetylenic hydrogen), 2.42 – 2.33 (m, 4H, CH × 2), 1.40 (s, 9H, CH × 3). ESI-MS:
2
3
+
m/z = 225 [M+H] .
4
.1.11. General procedure D for synthesis of aromatic propargylamines (13i-k)
1
7

ACCEPTED MANUSCRIPT
To a round-bottomed flask was added 3-bromopropyne (1.0 equiv), phenylamine
1.0 equiv), K CO (2.0 equiv) and acetone. The mixture was stirred at room
(
2
3
temperature for 24 h. The solvent was removed and the residue was partitioned
between water and CH Cl . The organic layer was dried over magnesium sulfate and
2
2
under reduced pressure. The crude product was further purified by silica gel column
chromatography to give the desired propargylamine.
4
.1.11.1. N-(Prop-2-ynyl)aniline (13i)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol), aniline
(
394 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol) according to the general
2 3
synthesis procedure D to afford the title compound (183 mg, 1.39 mmol, 33% yield)
1
as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 7.11 (dd, J = 8.5, 7.5 Hz, 2H,
6
Ar-H), 6.64 (d, J = 8.5 Hz, 2H, Ar-H), 6.60 (t, J = 7.5 Hz, 1H, Ar-H), 5.94 (t, J = 6.0
Hz, 1H, NH), 3.85 (dd, J = 6.0, 2.5 Hz, 2H, CH ), 3.05 (t, J = 2.5 Hz, 1H, acetylenic
2
+
hydrogen). ESI-MS: m/z = 132 [M+H] .
4
.1.11.2. 4-Methoxy-N-(prop-2-ynyl)aniline (13j)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
p-anisidine (522 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol) according to the
2
3
general synthesis procedure D to afford the title compound (151 mg, 0.94 mmol, 22%
1
yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 6.75 (d, J = 9.0 Hz, 2H,
6
Ar-H), 6.61 (d, J = 9.0 Hz, 2H, Ar-H), 5.52 (t, J = 6.5 Hz, 1H, NH), 3.80 (dd, J = 6.5,
2
.5 Hz, 2H, CH ), 3.65 (s, 3H, OCH ), 3.02 (t, J = 2.5 Hz, 1H, acetylenic hydrogen).
2 3
+
ESI-MS: m/z = 162 [M+H] .
4
.1.11.3. 4-Fluoro-N-(prop-2-ynyl)aniline (13k)
This compound was prepared from 3-bromopropyne (500 mg, 4.24 mmol),
-fluoroaniline (471 mg, 4.24 mmol) and K CO (1.17 g, 8.48 mmol) according to the
4
2
3
general synthesis procedure D to afford the title compound (179 mg, 1.20 mmol, 28%
1
yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 6.96 (t, J = 9.0 Hz, 2H,
6
1
8

ACCEPTED MANUSCRIPT
Ar-H), 6.65 – 6.61 (m, 2H, Ar-H), 5.90 (t, J = 6.0 Hz, 1H, NH), 3.84 (dd, J = 6.0, 2.5
Hz, 2H, CH ), 3.05 (t, J = 2.5 Hz, 1H, acetylenic hydrogen). ESI-MS: m/z = 150
2
+
[
M+H] .
4
.1.12. General procedure E for sonogashira coupling reaction (14a-l)
-Bromo-4-iodoquinoline (12) (1.0 equiv), Pd(PPh ) Cl (0.1 equiv), CuI (0.15
6
3
2
2
equiv) and triethylamine were charged in a three-neck round bottom flask. The flask
was fitted with a N inlet adaptor and purged with N for 10 min. The solution of
2
2
alkyne (1.0 equiv) was then added via syringe and purged with N for another 10 min.
2
The reaction mixture was stirred at 50 °C for 5 h. After the completion of reaction, the
mixture was concentrated under reduced pressure and the residue was dissolved in
EtOAc, washed with 1 N NaOH and water, then the organic phase was dried over
magnesium sulfate. The crude product was purified by silica gel column
chromatography yielded the desired compound.
4
.1.12.1. 4-(6-Bromoquinolin-4-yl)but-3-yn-1-ol (14a)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and commercially available but-3-yn-1-ol (13a) (21 mg, 0.30 mmol) according
to the general synthesis procedure E to afford the title compound (58 mg, 0.21 mmol,
1
7
1
0% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ 8.88 (d, J = 4.5 Hz,
6
H, Ar-H), 8.38 (d, J = 2.0 Hz, 1H, Ar-H), 7.98 (d, J = 9.0 Hz, 1H, Ar-H), 7.93 (dd, J
=
9.0, 2.0 Hz, 1H, Ar-H), 7.61 (d, J = 4.5 Hz, 1H, Ar-H), 5.05 (t, J = 5.5 Hz, 1H, OH),
3
.69 (q, J = 6.5 Hz, 2H, CH ), 2.77 (t, J = 6.5 Hz, 2H, CH ). ESI-MS: m/z = 276
2
2
+
[
M+H] .
4
.1.12.2. 6-Bromo-4-(3-(piperidin-1-yl)prop-1-ynyl)quinoline (14b)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 1-(prop-2-ynyl)piperidine (13b) (37 mg, 0.30 mmol) according to the
general synthesis procedure E to afford the title compound (56 mg, 0.17 mmol, 57%
1
yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ 8.90 (d, J = 4.5 Hz, 1H,
6
1
9

ACCEPTED MANUSCRIPT
Ar-H), 8.38 (d, J = 2.0 Hz, 1H, Ar-H), 8.00 (d, J = 9.0 Hz, 1H, Ar-H), 7.94 (dd, J =
9
.0, 2.0 Hz, 1H, Ar-H), 7.66 (d, J = 4.5 Hz, 1H, Ar-H), 3.70 (s, 2H, CH ), 2.56 (m, 4H,
2
CH × 2), 1.58 – 1.52 (m, 4H, CH × 2), 1.42 – 1.37 (m, 2H, CH ). ESI-MS: m/z =
2
2
2
+
3
29 [M+H] .
4
.1.12.3. 6-Bromo-4-(3-(4-methylpiperazin-1-yl)prop-1-ynyl)quinoline (14c)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 1-methyl-4-(prop-2-ynyl)piperazine (13c) (41 mg, 0.30 mmol) according
to the general synthesis procedure E to afford the title compound (62 mg, 0.18 mmol,
1
6
0% yield) as a viscous oil. H NMR (500 MHz, CDCl ) δ 8.87 (d, J = 4.5 Hz, 1H,
3
Ar-H), 8.36 (d, J = 2.0 Hz, 1H, Ar-H), 7.98 (d, J = 9.0 Hz, 1H, Ar-H), 7.80 (dd, J =
9
.0, 2.0 Hz, 1H, Ar-H), 7.61 (d, J = 4.5 Hz, 1H, Ar-H), 3.76 (s, 2H, CH ), 3.01 – 2.85
2
+
(
m, 8H, CH × 4), 2.57 (s, 3H, CH ). ESI-MS: m/z = 344 [M+H] .
2
3
4
.1.12.4. 1-(3-(6-Bromoquinolin-4-yl)prop-2-ynyl)piperidin-4-ol (14d)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 1-(prop-2-ynyl)piperidin-4-ol (13d) (42 mg, 0.30 mmol) according to the
general synthesis procedure E to afford the title compound (58 mg, 0.17 mmol, 57%
1
yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ 8.92 (d, J = 4.5 Hz, 1H,
6
Ar-H), 8.38 (d, J = 2.0 Hz, 1H, Ar-H), 8.03 (d, J = 9.0 Hz, 1H, Ar-H), 7.97 (dd, J =
9
3
1
.0, 2.0 Hz, 1H, Ar-H), 7.68 (d, J = 4.5 Hz, 1H, Ar-H), 4.61 (d, J = 4.5 Hz, 1H, OH),
.74 (s, 2H, CH ), 3.50 (m, 1H, CH), 2.86 (m, 2H, CH ), 2.44 – 2.30 (m, 2H, CH ),
2
2
2
+
.77 (m, 2H, CH ), 1.47 (m, 2H, CH ). ESI-MS: m/z = 345 [M+H] .
2
2
4
.1.12.5. 1-(3-(6-Bromoquinolin-4-yl)prop-2-ynyl)piperidin-3-ol (14e)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 1-(prop-2-ynyl)piperidin-3-ol (13e) (42 mg, 0.30 mmol) according to the
general synthesis procedure E to afford the title compound (51 mg, 0.15 mmol, 50%
1
yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ 8.92 (d, J = 4.5 Hz, 1H,
6
Ar-H), 8.38 (d, J = 2.0 Hz, 1H, Ar-H), 8.02 (d, J = 9.0 Hz, 1H, Ar-H), 7.96 (dd, J =
2
0

ACCEPTED MANUSCRIPT
9
.0, 2.0 Hz, 1H, Ar-H), 7.67 (d, J = 4.5 Hz, 1H, Ar-H), 4.72 (s, 1H, OH), 3.76 (s, 2H,
CH ), 3.56 (m, 1H, CH), 2.94 (dd, J = 10.0, 4.0 Hz, 1H, CH ), 2.77 (d, J = 11.0 Hz,
2
2
1
H, CH ), 2.24 (td, J = 11.0, 3.0 Hz, 1H, CH ), 2.10 (t, J = 10.0 Hz, 1H, CH ), 1.85 –
2 2 2
1
.79 (m, 1H, CH ), 1.74 – 1.67 (m, 1H, CH ), 1.52 – 1.44 (m, 1H, CH ), 1.15 – 1.07
2
2
2
+
(
m, 1H, CH ). ESI-MS: m/z = 345 [M+H] .
2
4
.1.12.6. 2-(3-(6-Bromoquinolin-4-yl)prop-2-ynyloxy)ethanol (14f)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and commercially available 2-(prop-2-ynyloxy)ethanol (13f) (30 mg, 0.30
mmol) according to the general synthesis procedure E to afford the title compound
1
(
55 mg, 0.18 mmol, 60% yield) as an off-white solid. H NMR (500 MHz, DMSO-d )
6
δ 8.92 (d, J = 4.5 Hz, 1H, Ar-H), 8.32 (d, J = 2.0 Hz, 1H, Ar-H), 8.01 (d, J = 9.0 Hz,
H, Ar-H), 7.95 (dd, J = 9.0, 2.0 Hz, 1H, Ar-H), 7.69 (d, J = 4.5 Hz, 1H, Ar-H), 4.73
t, J = 5.5 Hz, 1H, OH), 4.61 (s, 2H, CH ), 3.64 – 3.61 (m, 2H, CH ), 3.60 – 3.56 (m,
1
(
2
2
+
2
H, CH ). ESI-MS: m/z = 306 [M+H] .
2
4
.1.12.7. 2-((3-(6-Bromoquinolin-4-yl)prop-2-ynyl)(methyl)amino)ethanol (14g)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 2-(methyl(prop-2-ynyl)amino)ethanol (13g) (34 mg, 0.30 mmol)
according to the general synthesis procedure E to afford the title compound (51 mg,
1
0
4
7
.16 mmol, 53% yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 8.90 (d, J =
6
.5 Hz, 1H, Ar-H), 8.36 (d, J = 2.0 Hz, 1H, Ar-H), 8.00 (d, J = 9.0 Hz, 1H, Ar-H),
.95 (dd, J = 9.0, 2.0 Hz, 1H, Ar-H), 7.66 (d, J = 4.5 Hz, 1H, Ar-H), 4.49 (t, J = 5.5
Hz, 1H, OH), 3.78 (s, 2H, CH ), 3.53 (q, J = 6.0 Hz, 2H, CH ), 2.58 (t, J = 6.0 Hz, 2H,
2
2
+
CH ), 2.37 (s, 3H, CH ). ESI-MS: m/z = 319 [M+H] .
2
3
4
.1.12.8. 3-((3-(6-Bromoquinolin-4-yl)prop-2-ynyl)(methyl)amino)propane-1,2-diol
14h)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 3-(methyl(prop-2-ynyl)amino)propane-1,2-diol (13h) (43 mg, 0.30 mmol)
(
2
1

ACCEPTED MANUSCRIPT
according to the general synthesis procedure E to afford the title compound (66 mg,
1
0
4
7
.19 mmol, 63% yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 8.90 (d, J =
6
.5 Hz, 1H, Ar-H), 8.37 (d, J = 2.0 Hz, 1H, Ar-H), 8.00 (d, J = 9.0 Hz, 1H, Ar-H),
.94 (dd, J = 9.0, 2.0 Hz, 1H, Ar-H), 7.66 (d, J = 4.5 Hz, 1H, Ar-H), 4.50 (d, J = 4.5
Hz, 2H, OH × 2), 3.80 (s, 2H, CH ), 3.62 (m, 1H, CH), 3.40 – 3.33 (m, 2H, CH ),
2
2
2
.57 (dd, J = 12.5, 5.0 Hz, 1H, CH ), 2.44 (dd, J = 12.5, 7.0 Hz, 1H, CH ), 2.39 (s, 3H,
2 2
+
CH ). ESI-MS: m/z = 349 [M+H] .
3
4
.1.12.9. N-(3-(6-Bromoquinolin-4-yl)prop-2-ynyl)aniline (14i)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and N-(prop-2-ynyl)aniline (13i) (39 mg, 0.30 mmol) according to the general
synthesis procedure E to afford the title compound (60 mg, 0.18 mmol, 60% yield) as
1
a viscous oil. H NMR (500 MHz, DMSO-d ) δ 8.89 (d, J = 4.5 Hz, 1H, Ar-H), 8.24
6
(d, J = 2.0 Hz, 1H, Ar-H), 7.98 (d, J = 9.0 Hz, 1H, Ar-H), 7.92 (dd, J = 9.0, 2.0 Hz,
1
H, Ar-H), 7.60 (d, J = 4.5 Hz, 1H, Ar-H), 7.19 (dd, J = 9.5, 9.0 Hz, 2H, Ar-H ), 6.80
(d, J = 9.5 Hz, 2H, Ar-H), 6.66 (t, J = 9.0 Hz, 1H, Ar-H), 6.21 (t, J = 6.5 Hz, 1H, NH),
+
4
.35 (d, J = 6.5 Hz, 2H, CH ). ESI-MS: m/z = 337 [M+H] .
2
4
.1.12.10. N-(3-(6-Bromoquinolin-4-yl)prop-2-ynyl)-4-methoxyaniline (14j)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 4-methoxy-N-(prop-2-ynyl)aniline (13j) (48 mg, 0.30 mmol) according to
the general synthesis procedure E to afford the title compound (52 mg, 0.14 mmol,
1
4
8
7
7% yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 8.87 (s, 1H, Ar-H),
6
.16 (s, 1H, Ar-H), 7.95 (d, J = 9.0 Hz, 1H, Ar-H), 7.89 (d, J = 9.0 Hz, 1H, Ar-H),
.57 (d, J = 4.0 Hz, 1H, Ar-H), 6.80 (d, J = 8.5 Hz, 2H, Ar-H), 6.75 (d, J = 8.5 Hz, 2H,
Ar-H), 5.79 (t, J = 6.5 Hz, 1H, NH), 4.27 (d, J = 6.5 Hz, 2H, CH ), 3.64 (s, 3H,
2
+
OCH ). ESI-MS: m/z = 367 [M+H] .
3
4
.1.12.11. N-(3-(6-Bromoquinolin-4-yl)prop-2-ynyl)-4-fluoroaniline (14k)
2
2

ACCEPTED MANUSCRIPT
This compound was prepared from 6-bromo-4-iodoquinoline (12) (100 mg, 0.30
mmol) and 4-fluoro-N-(prop-2-ynyl)aniline (13k) (45 mg, 0.30 mmol) according to
the general synthesis procedure E to afford the title compound (63 mg, 0.18 mmol,
1
6
0% yield) as a viscous oil. H NMR (500 MHz, DMSO-d ) δ 8.87 (d, J = 4.5 Hz, 1H,
6
Ar-H), 8.13 (d, J = 2.0 Hz, 1H, Ar-H), 7.95 (d, J = 9.0 Hz, 1H, Ar-H), 7.89 (dd, J =
9
6
.0, 2.0 Hz, 1H, Ar-H), 7.57 (d, J = 4.5 Hz, 1H, Ar-H), 7.01 (t, J = 9.0 Hz, 2H, Ar-H ),
.84 – 6.75 (m, 2H, Ar-H), 6.15 (t, J = 6.5 Hz, 1H, NH), 4.31 (d, J = 6.5 Hz, 2H, CH2).
+
ESI-MS: m/z = 355 [M+H] .
4
4
.1.12.12.
Tert-butyl
-(3-(6-bromoquinolin-4-yl)prop-2-ynyl)piperazine-1-carboxylate (14l)
This compound was prepared from 6-bromo-4-iodoquinoline (12) (400 mg, 1.20
mmol) and tert-butyl 4-(prop-2-ynyl)piperazine-1-carboxylate (13l) (269 mg, 1.20
mmol) according to the general synthesis procedure E to afford the title compound
1
(
224 mg, 0.52 mmol, 43% yield) as an off-white solid. H NMR (500 MHz, DMSO-d )
6
δ 8.93 (d, J = 4.5 Hz, 1H, Ar-H), 8.36 (d, J = 2.0 Hz, 1H, Ar-H), 8.03 (d, J = 9.0 Hz,
H, Ar-H), 7.97 (dd, J = 9.0, 2.0 Hz, 1H, Ar-H), 7.70 (d, J = 4.5 Hz, 1H, Ar-H), 3.82
s, 2H, CH ), 3.40 (m, 4H, CH × 2), 2.60 – 2.55 (m, 4H, CH × 2), 1.40 (s, 9H, CH
1
(
2
2
2
3
+
×
3). ESI-MS: m/z = 430 [M+H] .
4
.1.13. General procedure F for suzuki coupling reaction (15a-n, 19a-e)
To a three-neck round bottom flask was added a halide (1.0 equiv), a boronic ester
1.0 equiv), Pd(dppf) Cl (0.1 equiv) and K CO (3.0 equiv) in dioxane/H O (3/1).
(
2
2
2
3
2
The flask was fitted with a N inlet adaptor and purged with N for 15 min. The
2
2
reaction mixture was then sealed under an atmosphere of N and stirred at 100 °C for
2
1
0 h. The crude mixture was concentrated under reduced pressure and the residue was
dissolved in CH Cl , washed with water twice, then the organic phase was dried over
2
2
magnesium sulfate. The crude product was purified by silica gel column
chromatography to give the desired target compound.
2
3

ACCEPTED MANUSCRIPT
4
2
.1.13.1.
,4-Difluoro-N-(5-(4-(4-hydroxybut-1-ynyl)quinolin-6-yl)-2-methoxypyridin-3-yl)benz
enesulfonamide (15a)
This compound was prepared from 14a (33 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (25
1
mg, 0.051 mmol, 43% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.36 (s, 1H, NH), 8.85 (d, J = 4.5 Hz, 1H, Ar-H), 8.48 (d, J = 2.5 Hz, 1H, Ar-H),
.39 (d, J = 2.0 Hz, 1H, Ar-H), 8.12 (d, J = 9.0 Hz, 1H, Ar-H), 8.07 (dd, J = 9.0, 2.0
8
Hz, 1H, Ar-H), 8.03 (d, J = 2.5 Hz, 1H, Ar-H), 7.77 (m, 1H, Ar-H), 7.59 (d, J = 4.5
Hz, 1H, Ar-H), 7.56(m, 1H, Ar-H), 7.20 (m, 1H, Ar-H), 5.04 (t, J = 5.5 Hz, 1H, OH),
13
3
.72 (q, J = 6.5 Hz, 2H, CH ), 3.68 (s, 3H, OCH ), 2.78 (t, J = 6.5 Hz, 2H, CH ). C
2 3 2
NMR (100 MHz, DMSO-d ) δ 165.06 (dd, JC-F = 252.8, 11.2 Hz), 159.36 (dd, JC-F
=
=
6
2
1
1
1
55.9, 13.4 Hz), 157.59, 150.31, 146.88, 142.62, 135.02, 133.96, 131.83 (d, JC-F
1.2 Hz), 130.45, 129.41, 128.75, 128.73, 127.53, 125.01 (dd, JC-F = 14.2, 3.3 Hz),
24.04, 122.42, 119.82, 111.85 (dd, JC-F = 22.8, 3.8 Hz), 105.81 (t, JC-F = 25.8 Hz),
+
00.20, 76.65, 59.48, 53.49, 23.69. ESI-MS: m/z = 496 [M+H] .
4
2
3
.1.13.2.
,4-Difluoro-N-(2-methoxy-5-(4-(3-(piperidin-1-yl)prop-1-ynyl)quinolin-6-yl)pyridin-
-yl)benzenesulfonamide (15b)
This compound was prepared from 14b (39 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (16
1
mg, 0.029 mmol, 24% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.38 (s, 1H, NH), 8.87 (d, J = 4.5 Hz, 1H, Ar-H), 8.42 (d, J = 2.5 Hz, 1H, Ar-H),
.39 (d, J = 2.0 Hz, 1H, Ar-H), 8.14 (d, J = 9.0 Hz, 1H, Ar-H), 8.09 (dd, J = 9.0, 2.0
8
Hz, 1H, Ar-H), 7.99 (d, J = 2.5 Hz, 1H, Ar-H), 7.74 (m, 1H, Ar-H), 7.64 (d, J = 4.5
Hz, 1H, Ar-H), 7.53 (m, 1H, Ar-H), 7.17 (m, 1H, Ar-H), 3.74 (s, 2H, CH ), 3.66 (s,
2
3
H, OCH ), 2.61 (m, 4H, CH × 2), 1.59 – 1.52 (m, 4H, CH × 2), 1.35 (m, 2H, CH ).
3 2 2 2
1
3
C NMR (100 MHz, DMSO-d ) δ 164.98 (dd, J = 251.7, 11.0 Hz), 159.36 (dd, JC-F
6
C-F
=
254.3, 11.8 Hz), 157.72, 150.35, 146.88, 142.41, 135.31, 133.76, 131.77 (d, JC-F =
2
4

ACCEPTED MANUSCRIPT
1
1
5
0.0 Hz), 130.55, 128.82, 128.71, 127.28, 125.18 (d, JC-F = 13.6 Hz), 124.21, 122.31,
20.27, 111.75 (dd, JC-F = 21.9, 3.8 Hz), 105.73 (t, JC-F = 25.6 Hz), 96.66, 80.48,
+
3.42, 52.63, 47.68, 25.37, 23.39. ESI-MS: m/z = 549 [M+H] .
4
2
.1.13.3.
,4-Difluoro-N-(2-methoxy-5-(4-(3-(4-methylpiperazin-1-yl)prop-1-ynyl)quinolin-6-yl
)pyridin-3-yl)benzenesulfonamide (15c)
This compound was prepared from 14c (41 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (21
1
mg, 0.037 mmol, 31% yield) as an off-white solid. H NMR (500 MHz, CDCl ) δ 8.85
3
(d, J = 4.5 Hz, 1H, Ar-H), 8.36 (s, 1H, Ar-H), 8.26 (s, 1H, Ar-H), 8.17 (d, J = 9.0 Hz,
1
7
2
1
1
1
H, Ar-H), 8.12 (s, 1H, Ar-H), 7.88 (m, 2H, Ar-H), 7.53 (d, J = 4.5 Hz, 1H, Ar-H),
.00 – 6.90 (m, 2H, Ar-H), 3.95 (s, 3H, OCH ), 3.75 (s, 2H, CH ), 2.82 (s, 4H, CH ×
3
2
2
13
), 2.62 (s, 4H, CH × 2), 2.36 (s, 3H, CH ). C NMR (125 MHz, DMSO-d ) δ
2
3
6
65.12 (dd, JC-F = 251.0, 11.0 Hz), 159.81 (dd, JC-F = 255.5, 13.5 Hz), 158.14, 150.67,
47.33, 141.13, 136.13, 132.53, 132.15 (d, JC-F = 11.1 Hz), 130.98, 129.25, 129.02,
28.99, 127.78, 126.73 (dd, JC-F = 15.6, 4.5 Hz), 124.75, 123.06, 122.50, 112.02 (dd,
JC-F = 21.6, 3.1 Hz), 106.08 (t, JC-F = 26.1 Hz), 96.68, 81.08, 54.63, 53.79, 51.13,
+
4
7.24, 45.41. ESI-MS: m/z = 564 [M+H] .
4
2
.1.13.4.
,4-Difluoro-N-(5-(4-(3-(4-hydroxypiperidin-1-yl)prop-1-ynyl)quinolin-6-yl)-2-metho
xypyridin-3-yl)benzenesulfonamide (15d)
This compound was prepared from 14d (41 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (18
1
mg, 0.032 mmol, 27% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.36 (s, 1H, NH), 8.89 (d, J = 4.5 Hz, 1H, Ar-H), 8.43 (d, J = 2.5 Hz, 1H, Ar-H),
.38 (d, J = 2.0 Hz, 1H, Ar-H), 8.16 (d, J = 9.0 Hz, 1H, Ar-H), 8.11 (dd, J = 9.0, 2.0
8
Hz, 1H, Ar-H), 8.01 (d, J = 2.5 Hz, 1H, Ar-H), 7.78 (m, 1H, Ar-H), 7.67 (d, J = 4.5
Hz, 1H, Ar-H), 7.55 (m, 1H, Ar-H), 7.21 (m, 1H, Ar-H), 4.52 (s, 1H, OH), 3.76 (s, 2H,
2
5

ACCEPTED MANUSCRIPT
CH ), 3.70 (s, 3H, OCH ), 3.45 (m, 1H, CH), 2.87 (m, 2H, CH ), 2.42 (m, 2H, CH ),
2
3
2
2
13
1
.79 (m, 2H, CH ), 1.46 (m, 2H, CH ). C NMR (100 MHz, DMSO-d ) δ 164.98 (dd,
2 2 6
JC-F = 251.6, 9.6 Hz), 159.41 (dd, JC-F = 267.8, 13.4 Hz), 157.68, 150.34, 146.86,
1
1
2
3
42.37, 135.28, 133.75, 131.77 (d, JC-F = 10.3 Hz), 130.54, 128.81, 128.67, 128.64,
27.28, 125.16 (dd, JC-F = 13.5, 4.2 Hz), 124.31, 122.27, 120.23, 111.81 (dd, JC-F
=
1.2, 3.8 Hz), 105.77 (t, JC-F = 25.7 Hz), 96.63, 80.34, 63.7, 53.47, 49.75, 47.03,
+
4.19. ESI-MS: m/z = 565 [M+H] .
4
2
.1.13.5.
,4-Difluoro-N-(5-(4-(3-(3-hydroxypiperidin-1-yl)prop-1-ynyl)quinolin-6-yl)-2-metho
xypyridin-3-yl)benzenesulfonamide (15e)
This compound was prepared from 14e (41 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (15
1
mg, 0.027 mmol, 23% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.38 (s, 1H, NH), 8.90 (d, J = 4.5 Hz, 1H, Ar-H), 8.47 (d, J = 2.5 Hz, 1H, Ar-H),
.40 (d, J = 2.0 Hz, 1H, Ar-H), 8.17 (d, J = 9.0 Hz, 1H, Ar-H), 8.12 (dd, J = 9.0, 2.0
8
Hz, 1H, Ar-H), 8.03 (d, J = 2.5 Hz, 1H, Ar-H), 7.78 (m, 1H, Ar-H), 7.68 (d, J = 4.5
Hz, 1H, Ar-H), 7.60 – 7.54 (m, 1H, Ar-H), 7.21 (td, J = 8.5, 2.0 Hz, 1H, Ar-H), 4.67
(
s, 1H, OH), 3.78 (d, J = 4.5 Hz, 2H, CH ), 3.69 (s, 3H, OCH ), 3.59 – 3.53 (m, 1H,
2 3
CH ), 2.96 – 2.91 (dd, J = 10.5, 3.5 Hz, 1H, CH ), 2.82 (m, 1H, CH ), 2.32 (t, J = 9.5
2
2
2
Hz, 1H, CH ), 2.14 (t, J = 9.5 Hz, 1H, CH ), 1.76 (m, 2H, CH ), 1.49 (m, 1H, CH ),
2
2
2
2
13
1
.08 (m, 1H, CH ). C NMR (125 MHz, DMSO-d ) δ 165.51 (dd, J = 251.8, 11.5
2 6 C-F
Hz), 159.86 (dd, JC-F = 255.8, 12.7 Hz), 158.18, 150.79, 147.37, 142.96, 135.78,
34.30, 132.30 (d, JC-F = 10.8 Hz), 131.03, 129.29, 129.22, 129.15, 127.76, 125.65 (d,
1
JC-F = 11.3 Hz), 124.78, 122.78, 120.62, 112.28 (dd, JC-F = 21.6, 2.8 Hz), 106.24 (t,
JC-F = 25.9 Hz), 96.92, 81.03, 66.41, 60.37, 53.93, 52.16, 47.68, 33.19, 23.34. ESI-MS:
+
m/z = 565 [M+H] .
4
2
.1.13.6.
,4-Difluoro-N-(5-(4-(3-(2-hydroxyethoxy)prop-1-ynyl)quinolin-6-yl)-2-methoxypyridi
2
6

ACCEPTED MANUSCRIPT
n-3-yl)benzenesulfonamide (15f)
This compound was prepared from 14f (37 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (25
1
mg, 0.048 mmol, 40% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.36 (s, 1H, NH), 8.90 (d, J = 4.5 Hz, 1H, Ar-H), 8.47 (d, J = 2.5 Hz, 1H, Ar-H),
.32 (d, J = 2.0 Hz, 1H, Ar-H), 8.15 (d, J = 9.0 Hz, 1H, Ar-H), 8.09 (dd, J = 9.0, 2.0
8
Hz, 1H, Ar-H), 8.01 (d, J = 2.5 Hz, 1H, Ar-H), 7.76 (m, 1H, Ar-H), 7.68 (d, J = 4.5
Hz, 1H, Ar-H), 7.59 – 7.53 (m, 1H, Ar-H), 7.20 (m, 1H, Ar-H), 4.69 (s, 1H, OH), 4.63
1
3
(
s, 2H, CH ), 3.67 (s, 3H, OCH ), 3.66 – 3.62 (m, 2H, CH ), 3.58 (m, 2H, CH ). C
2 3 2 2
NMR (100 MHz, DMSO-d ) δ 165.07 (dd, JC-F = 254.2, 12.8 Hz), 159.36 (dd, JC-F
=
=
6
2
1
1
8
55.7, 12.9 Hz), 157.66, 150.32, 146.87, 142.72, 135.39, 134.05, 131.83 (d, JC-F
0.7 Hz), 130.55, 128.98, 128.70, 128.10, 127.09, 125.01 (d, JC-F = 13.6 Hz), 124.49,
22.15, 119.86, 111.85 (dd, JC-F = 21.8, 2.7 Hz), 105.80 (t, JC-F = 27.0 Hz), 96.79,
+
0.86, 71.63, 60.07, 58.24, 53.48. ESI-MS: m/z = 526 [M+H] .
4
2
.1.13.7.
,4-Difluoro-N-(5-(4-(3-((2-hydroxyethyl)(methyl)amino)prop-1-ynyl)quinolin-6-yl)-2
-methoxypyridin-3-yl)benzenesulfonamide (15g)
This compound was prepared from 14g (38 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (28
1
mg, 0.052 mmol, 43% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.34 (s, 1H, NH), 8.88 (d, J = 4.5 Hz, 1H, Ar-H), 8.45 (d, J = 2.5 Hz, 1H, Ar-H),
.38 (d, J = 2.0 Hz, 1H, Ar-H), 8.14 (d, J = 9.0 Hz, 1H, Ar-H), 8.09 (dd, J = 9.0, 2.0
8
Hz, 1H, Ar-H), 8.01 (d, J = 2.5 Hz, 1H, Ar-H), 7.75 (m, 1H, Ar-H), 7.66 (d, J = 4.5
Hz, 1H, Ar-H), 7.55 (m, 1H, Ar-H), 7.19 (m, 1H, Ar-H), 4.47 (s, 1H, OH), 3.83 (s, 2H,
CH ), 3.67 (s, 3H, OCH ), 3.54 (t, J = 6.0 Hz, 2H, CH ), 2.62 (t, J = 6.0 Hz, 2H, CH ),
2
3
2
2
13
2
.42 (s, 3H, CH ). C NMR (100 MHz, DMSO-d ) δ 165.05 (dd, J = 251.7, 9.6
3 6 C-F
Hz), 159.52 (dd, JC-F = 255.6, 8.6 Hz), 157.78, 150.40, 146.94, 142.45, 135.32,
33.91, 131.82 (d, JC-F = 10.6 Hz), 130.61, 128.90, 128.76, 128.72, 127.32, 125.27
dd, JC-F = 13.8, 3.6 Hz), 124.45, 122.25, 120.30, 111.85 (dd, JC-F = 22.0, 2.9 Hz),
1
(
2
7

ACCEPTED MANUSCRIPT
1
05.82 (t, JC-F = 25.3 Hz), 96.45, 80.60, 58.96, 57.78, 54.93, 46.43, 42.08. ESI-MS:
+
m/z = 539 [M+H] .
4
.1.13.8.
N-(5-(4-(3-((2,3-Dihydroxypropyl)(methyl)amino)prop-1-ynyl)quinolin-6-yl)-2-metho
xypyridin-3-yl)-2,4-difluorobenzenesulfonamide (15h)
This compound was prepared from 14h (42 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (33
1
mg, 0.058 mmol, 48% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.32 (s, 1H, NH), 8.88 (s, 1H, Ar-H), 8.44 (s, 1H, Ar-H), 8.38 (s, 1H, Ar-H), 8.14 (d,
J = 8.5 Hz, 1H, Ar-H), 8.08 (d, J = 7.5 Hz, 1H, Ar-H), 8.00 (s, 1H, Ar-H), 7.75 (m,
H, Ar-H), 7.66 (s, 1H, Ar-H), 7.55 (m, 1H, Ar-H), 7.19 (m, 1H, Ar-H), 4.48 (s, 2H,
OH × 2), 3.84 (s, 2H, CH ), 3.67 (s, 3H, OCH ), 3.63 (s, 1H, CH), 3.35 (m, 2H, CH ),
1
2
3
2
1
3
2
1
1
1
.59 (m, 1H, CH ), 2.41 (m, 4H, CH + CH ). C NMR (125 MHz, DMSO-d ) δ
2 3 2 6
65.49 (dd, JC-F = 252.6, 11.6 Hz), 159.85 (dd, JC-F = 256.0, 13.6 Hz), 158.19, 150.79,
47.39, 142.84, 135.78, 134.26, 132.27 (d, JC-F = 10.6 Hz), 131.03, 129.33, 129.24,
29.19, 127.77, 125.73 (dd, JC-F = 14.9, 4.0 Hz), 124.91, 122.73, 120.76, 112.28 (dd,
JC-F = 22.0, 2.9 Hz), 106.24 (t, JC-F = 25.9 Hz), 96.96, 81.03, 69.90, 65.00, 59.34,
+
5
3.95, 47.26, 42.89. ESI-MS: m/z = 569 [M+H] .
4
2
.1.13.9.
,4-Difluoro-N-(2-methoxy-5-(4-(3-(phenylamino)prop-1-ynyl)quinolin-6-yl)pyridin-3
-yl)benzenesulfonamide (15i)
This compound was prepared from 14i (40 mg, 0.12 mmol) and 5a (50 mg, 0.12
mmol) according to the general synthesis procedure F to afford the title compound (18
1
mg, 0.032 mmol, 27% yield) as an off-white solid. H NMR (500 MHz, DMSO-d ) δ
6
1
0.39 (s, 1H, NH), 8.87 (d, J = 4.5 Hz, 1H, Ar-H), 8.36 (d, J = 2.5 Hz, 1H, Ar-H),
.25 (d, J = 2.0 Hz, 1H, Ar-H), 8.13 (d, J = 9.0 Hz, 1H, Ar-H), 8.05 (dd, J = 9.0, 2.0
8
Hz, 1H, Ar-H), 8.00 (d, J = 2.5 Hz, 1H, Ar-H), 7.78 (m, 1H, Ar-H), 7.60 (d, J = 4.5
Hz, 1H, Ar-H), 7.59 (m, 1H, Ar-H), 7.24 – 7.21 (m, 1H, Ar-H), 7.09 (m, 2H, Ar-H),
2
8