Functional induction of P-glycoprotein efflux pump by phenyl benzenesulfonamides: Synthesis and biological evaluation of T0901317 analogs

Article abstract of DOI:10.1016/j.ejmech.2016.07.005

N-(2,2,2-Trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide (T0901317, 6) is a potent activator of pregnane-X-receptor (PXR), which is a nuclear receptor controlling P-gp expression. Herein, we aimed to inv

Full text of DOI:10.1016/j.ejmech.2016.07.005

Accepted Manuscript
Functional induction of P-glycoprotein efflux pump by phenyl benzenesulfonamides:
Synthesis and biological evaluation of T0901317 analogs
Anil K. Padala, Abubakar Wani, Ram A. Vishwakarma, Ajay Kumar, Sandip B.
Bharate
PII:
S0223-5234(16)30552-9
10.1016/j.ejmech.2016.07.005
EJMECH 8723
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 9 March 2016
Revised Date: 17 June 2016
Accepted Date: 5 July 2016
Please cite this article as: A.K. Padala, A. Wani, R.A. Vishwakarma, A. Kumar, S.B. Bharate, Functional
induction of P-glycoprotein efflux pump by phenyl benzenesulfonamides: Synthesis and biological
evaluation of T0901317 analogs, European Journal of Medicinal Chemistry (2016), doi: 10.1016/
j.ejmech.2016.07.005.
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ACCEPTED MANUSCRIPT
GRAPHICAL ABSTRACT
O
O
O
CF3
S
N
HO
F3C
O
(Potent P-gp inducer)
Non-toxic to LS-180 cells
Non-toxic to SH-SH5Y cells
N
N
N
O
S
N
40 analogs
F3C
T0901317 (6)
26e
F3C
We report P-gp induction activity of a PXR activator T0901317 and its structure-activity
relationship. Analogs displaying promising P-gp induction have been identified.

ACCEPTED MANUSCRIPT
Functional induction of P-glycoprotein efflux pump by phenyl
benzenesulfonamides: Synthesis and biological evaluation of
T0901317 analogs
a,b$
c$
a,b
,c
Anil K. Padala,
Abubakar Wani, Ram A. Vishwakarma, Ajay Kumar,* and Sandip B.
,a,b
Bharate*
a
Medicinal Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Canal Road,
Jammu-180001, India.
b
Academy of Scientific & Innovative Research (AcSIR), CSIR-Indian Institute of Integrative
Medicine, Canal Road, Jammu-180001, India
c
Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road,
Jammu-180001, India.
$
A.K.P. and A.W. equally contributed for first authorship.
*
Corresponding authors
E-mail: sbharate@iiim.ac.in (S.B.B.),
ajaykmahajan@hotmail.com (AK)
Tel. +91-191-2585006-13, Ext. 345
1

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ABSTRACT.
N-(2,2,2-Trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-
benzenesulfonamide (T0901317, 6) is a potent activator of pregnane-X-receptor (PXR), which is
a nuclear receptor controlling P-gp expression. Herein, we aimed to investigate P-gp induction
activity of T0901317 and establish its structure-activity relationship. T0901317 along with a
series of N-triazolyl-methylene-linked benzenesulfonamides were synthesized and screened for
P-gp induction activity using a rhodamine-123 based efflux assay in the P-gp overexpressing
human adenocarcinoma LS-180 cells, wherein several compounds showed potent P-gp induction
activity at 5 µM. Treatment with benzene sulphonamides led to the decrease in intracellular
accumulation of a fluorescent P-gp substrate rhodamine-123 up to 48% (control 100%). In the
western-blot studies, T0901317 (6) and its triazole linked analog 26e at 5 µM displayed
induction of P-gp expression in LS180 cells. These compounds were non-toxic in LS-180 and
human neuroblastoma SH-SY5Y cells (IC50 > 50 µM). The compound 26e showed significant P-
gp induction even at 0.3 µM, indicating an excellent therapeutic window. These results clearly
indicate promise of this class of compounds as potential agents to enhance amyloid-β clearance
in Alzheimers patients.
Keywords.
T0901317; benzene sulfonamides; PXR, LS180; P-gp induction; Alzheimer’s disease
2

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1
. INTRODUCTION
The functional induction of ABC transporter efflux pump P-gp has been reported to be beneficial
to efflux out toxins from the body. Thus, this strategy has been strategically explored as a
potential treatment in numerous diseases wherein the excessive accumulation of toxic/ mis-
folded proteins is the key pathological event. The most deadly and incurable disease involving
accumulation of such mis-folded proteins is Alzheimer’s disease, wherein there is an increased
accumulation of Aβ plaques inside the brain [1]. Numerous studies have been published on
discovery of small molecule inducers of P-gp efflux pump for increasing amyloid-β clearance
across BBB [2-4]. Examples of P-gp inducers include rifampicin (1) [5], oleocanthal (2) [6, 7],
fascaplysin (3) [8], 4-arylquinoline derivative 4 [9] and a colupulone derivative 5 [10].
Figure 1.
The expression of P-gp is controlled by the nuclear receptors PXR [11-14] and LXR [15]. This
might be the possible reason for the substrate overlapping of PXR/ LXR and P-gp. Thus, there is
an opportunity to discover newer P-gp inducer leads from known PXR activators. T0901317 is a
potent activator of liver X receptor (LXR) and pregnane X receptor (PXR) [16]. Recently,
Gosselet’s group [15] have reported that T0901317 also induces P-gp expression. However, till
date, no medicinal chemistry efforts have been reported on this scaffold for P-gp induction
activity. Thus, in continuation to our efforts in this area [10], herein we investigated P-gp
induction potential of T0901317 (6) and established its structure-activity relationship. T0901317
along with a series of analogs were synthesized and evaluated for their P-gp induction activity
using in-vitro rhodamine 123 efflux assay in LS-180 cells. The functional induction of P-gp by
these compounds has also been demonstrated by western-blot analysis.
3

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2
2
. RESULTS AND DISCUSSION
.1. Synthesis of T0901317 and its analogs.
The synthesis of T0901317 (6) is depicted in Scheme 1. Treatment of aniline (7) with
hexafluoroacetone trihydrate (8) in the presence of 4-methylbenzenesulfonic acid gave 2-(4-
amino-phenyl)-1,1,1,3,3,3-hexafluoro-propan-2-ol (9) [17] in 80% yield. Compound 9 on
treatment with benzenesulfonyl chloride (10) produced N-(4-(1,1,1,3,3,3-hexafluoro-2-
hydroxypropan-2-yl)phenyl)benzenesulfonamide (11). Treatment of compound 11 with 2,2,2-
trifluoroethyl trifluoromethylsulfonate (12) yielded T-0901317 (6) in 80% yield. Next, the
treatment of sulfonamide 11 with propargyl bromide (13) yielded dipropargylated compound 14.
Further, in order to prepare mono-propargylated analog 18, first the compound 11 was treated
with PMB chloride, followed by treatment with propargyl bromide and finally deprotection of
PMB, resulting in formation of 18. Next, the treatment of N-propargyl compound 18 with
substituted aryl azides 19a-o under Click reaction conditions, produced triazoles 20a-o in 70-
9
0% yield [18] (Scheme 1).
Scheme 1.
The treatment of commercially available 4-aminoacetophenone (21) with benzenesulfonyl
chloride (10) gave sulfonamide 22. The treatment of 22 with 2,2,2-trifluoroethyl
trifluoromethylsulfonate (12) produced 23 in 80% yield. Next, the N-benzylated compound 24
was obtained by reaction of 22 with benzyl bromide. The reaction of benzenesulphonamide 22
with propargyl bromide yielded N-propargylated benzene sulfonamide 25 in 92% yield. Finally,
the click reaction of 25 with various substituted aryl azides 19a-i, n, p-x yielded triazole linked
benzenesulfonamides 26a-i, n, p-x in 70-90% yield (Scheme 2).
4

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1
It is noteworthy to mention that a significant difference in the H NMR chemical shift value of
N-CH of two triazole series 20a-o and 26a-x was noticed. In case of series, 26a-x, the N-CH
2
2
appeared between δ 4.98-5.08 ppm, whereas in series 20a-o, the N-CH signal appeared
2
comparatively at up-field shift at δ 4.78-4.82 ppm.
Scheme 2.
2
.2. P-gp induction activity.
All synthesized compounds along with T0901317 (6) were screened for P-gp induction activity
using a rhodamine-123 based efflux assay in P-gp over-expressing human colorectal
adenocarcinoma LS-180 cells. Rifampicin (10 ꢀM) was used as a positive control in this study.
T0901317 (6) showed promising P-gp induction as indicated by the decrease in % intracellular
accumulation of Rh123 in LS-180 cells (up to 64%) compared to that of control (100%). As
shown in the Table 1, on replacement of N-linked -CH CF of T0901317 (6) with ‘propargyl’
2
3
unit (compound 18), the P-gp induction activity was not affected. In order to ascertain that these
compounds are not toxic to the cells, the effect of these compounds on the cell viability of LS-
1
80 and neuroblastoma SH-SY5Y cells was assessed. T0901317 (6) was not toxic to both these
cells (IC50 > 50 µM). Similarly, the propargylated analog 18 was also found to be non-toxic
Table 1).
(
Table 1.
The P-gp induction results of triazolyl linked T0901317 analogs (wherein hexafluoropropan-2-ol
substituent on a phenyl ring was retained) are shown in Table 2. Most of the compounds from
this series showed P-gp induction at 5 µM, although none of the compounds showed better
5

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decrease in % intracellular accumulation of Rh-123 levels in LS180 cells. The analog 20j
possessing 2,6-di-F substituted phenyltriazole showed 65% intracellular accumulation of Rh 123
levels, which was similar to that of T0901317 (Table 2). Next, the in-vitro cytotoxicity of all
compounds from this series was assessed in LS-180 and SH-SY5Y cells at 50 µM (Table 2).
Some of the compounds were non-cytotoxic to both cells; however many showed toxicity either
to both cells or to at least one cell line (in these cases, the cell viability was reduced to less than
5
0%). These compounds include 20a (toxic to both cells), 20i (toxic to SH-SH5Y cells), 20m
(toxic to SH-SH5Y cells), 20g (toxic to SH-SH5Y cells), 20f (toxic to SH-SH5Y cells). In
general, it was observed that compounds were more toxic to SH-SH5Y cells than LS-180 cells.
From this series, the compounds 20j was selected for further studies, as it showed best P-gp
induction activity as well as it was not toxic to both cell lines.
Table 2.
The P-gp induction results of 4-acetylphenyl benzene sulfonamides 23, 24, 26a-i, 26n and 26p-x
(wherein hexafluoropropan-2-ol substituent on a phenyl ring was replaced with acetyl) are shown
in Table 3. Most of the compounds from this series showed P-gp induction at 5 µM, and many
of them displayed better decrease in % intracellular accumulation of Rh123 levels than
T0901317. The 4-acetylphenyl analog 23 wherein the N-CH CF was retained, showed similar
2
3
level of P-gp induction activity to that of T0901317. Furthermore, on replacement of N-CH
2 3
CF
with N-CH Ph (compound 24), a similar P-gp induction effect was observed. Amongst triazolyl
2
linked analogs, several compounds displayed better effect than T0901317. Particularly,
compounds 26a, 26e, and 26x showed < 55% intracellular Rh123 levels in LS-180 cells. It was
interesting to observe some effect of the substituent’s (present on triazolyl-phenyl ring) on the P-
gp induction activity. The presence of electron-withdrawing groups such as CF , NO on 2- or 4-
3
2
6

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position of phenyl ring (examples 26e, 26f, 26x, 26h, and 26i) showed potent P-gp induction
activity (Table 3). Finally, all compounds from this series were assessed for their effect on the
cell viability (at 50 µM). Unlike, in the earlier series (20a-o, shown in Table 2), the compounds
from this series 26a-x were better in terms of cytotoxicity (all of them are non-toxic) and
furthermore, the P-gp induction activity of this series of compounds was better than 20a-o series.
Table 3.
As the P-gp induction effect was observed at 5 µM and the safe concentration was observed to be
up to 50 µM; it indicated 10-fold therapeutic window. We further thought to investigate P-gp
induction activity at lower concentrations, for which we selected 2.5, 1.25, 0.6 and 0.3 µM as test
concentrations. The P-gp induction effect was analyzed for T0901317 (6) and analogs 20j, 20h,
2
6a, and 26e at these lower doses. Interestingly, all these compounds displayed significant P-gp
induction even at the lowest concentration tested (0.3 µM). The results of T0901317 (6), analog
0j and analog 26e are shown in Figure 2. The results of other compounds are provided in
supporting information.
2
Figure 2.
The promising P-gp induction activity of these compounds was further confirmed by western-
blot analysis in LS-180 cells. For western-blot study, T0901317 (6) along with compounds 26e
and 26x were selected. Upon exposure to compounds 6, 26e and 26x at 5 µM, the P-gp
expression in LS180 cells was increased significantly (Figure 3).
Figure 3.
7

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Next, we also investigated the ability of these compounds to overcome amyloid-β induced
neurotoxicity in neuronal cells. This experiment was performed in human neuroblastoma SH-
SY5Y cells after differentiation with 10 µM of retinoic acid for seven days. Compound treatment
was done 30 min prior to the treatment with 10 µg/ml amyloid-β for 48 h. Treatment with only
amyloid-β resulted in decrease in % cell viability to 68%. However, when amyloid-beta treated
cells were co-treated with test compounds 20j and 26e at 5 µM, the cell viability was retained
back to 75 and 79%. This indicated that these compounds possess neuroprotective effect in
neuronal cells.
3
. CONCLUSION
In conclusion, the present study has demonstrated that the phenyl benzene sulfonamide scaffold,
apart from its known PXR activation ability, also possess strong ability to induce P-gp
expression. Herein, for the first time, P-gp induction effect of T0901317 has been demonstrated.
The P-gp induction effect of benzene sulfonamide 26e at 0.3 µM with no toxicity up to 50 µM,
demonstrated excellent therapeutic window (>166-fold) for these compounds. As the expression
of P-gp plays major role in enhancing amyloid-β efflux across the BBB; these compounds have a
potential for exploring as anti-Alzheimer agents.
4
. EXPERIMENTAL SECTION
4
.1. General. All chemicals were obtained from Sigma-Aldrich Company and used as received.
1
13
H, and C NMR spectra were recorded on Brucker-Avance DPX FT-NMR 500 and 400 MHz
instruments. Chemical data for protons are reported in parts per million (ppm) downfield from
tetramethylsilane and are referenced to the residual proton in the NMR solvent (CDCl
3
, 7.26
1
3
ppm). Carbon nuclear magnetic resonance spectra ( C NMR) were recorded at 125 MHz or 100
8

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MHz: chemical data for carbons are reported in parts per million (ppm, δ scale) downfield from
tetramethylsilane and are referenced to the carbon resonance of the solvent (CDCl , 77.16 ppm).
3
ESI-MS and HRMS spectra were recorded on Agilent 1100 LC-Q-TOF and HRMS-6540-UHD
machines. IR spectra were recorded on Perkin-Elmer IR spectrophotometer. Melting points were
recorded on digital melting point apparatus.
4
.2. Preparation of 2-(4-aminophenyl)-1,1,1,3,3,3-hexafluoropropan-2-ol (9): To the
solution of aniline (7, 21.47 mmol) and pTSA (1.28 mmol) in xylene, was added
hexafluoroacetone trihydrate (8, 64.42 mmol) and reaction mixture was refluxed for five hours.
Reaction was allowed to cool to the room temperature, after which the product 9 was
crystallized, which was filtered and used for the next step.
1
4
.2.1. 2-(4-Aminophenyl)-1,1,1,3,3,3-hexafluoropropan-2-ol (9) [17]. H NMR (CDCl
3
, 400
),
MHz): δ 7.46 (d, J = 8.4 Hz, 2H, Ar-CH), 6.70 (d, J = 8.6 Hz, 2H, Ar-CH), 3.84 (brs, 2H, NH
2
3
.37 (s, 1H, OH).
.3. General procedure for preparation of 11 and 22. To the solution of substituted aniline (7
4
or 21, 10.0 mmol) in pyridine (1 M) at room temperature, aryl sulfonyl chloride (10, 10.5 mmol)
was added. The mixture was allowed to stir at room temperature overnight. After the complete
consumption of the substituted aniline (monitored by TLC), the reaction mixture was quenched
with water, and the aqueous layer was extracted with dichloromethane. The combined organic
layer was washed with aqueous copper sulfate. The organic layer was then dried over sodium
sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by
silica gel (#100-200) column chromatography using n-hexane: EtOAc as an eluent to get phenyl
benzene sulfonamides 11, 22.
9

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4
.3.1. N-(4-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)phenyl)benzenesulfonamide (11)
1
[
17]. H NMR (CDCl , 400 MHz): δ 7.83 (d, J = 7.6 Hz, 2H, Ar-CH), 7.58 (t, J = 9.5 Hz, 3H,
3
Ar-CH), 7.47 (t, J = 7.7 Hz, 2H, Ar-CH), 7.15 (d, J = 8.8 Hz, 2H, Ar-CH), 6.96 (brs, 1H, NH),
+
3
.50 (brs, 1H, OH); ESI-MS: m/z 397.90 [M-H] .
1
4
.3.2. N-(4-Acetylphenyl)benzenesulfonamide (22) [19]. H NMR (CDCl3, 400 MHz): δ 7.86-
7
.83 (m, 4H, Ar-CH), 7.57-7.54 (m, 1H, Ar-CH), 7.49-7.47 (m, 2H, Ar-CH), 7.15-7.13 (m, 2H,
Ar-CH), 7.00 (s, 1H, NH), 2.53 (s, 3H); IR (CHCl
3
): νmax 3436, 2921, 2089, 1661, 1601, 1510,
-1
+
1
447, 1404, 1360, 1334, 1272, 1239, 1160, 1121, 1091, 1020 cm ; ESI-MS: m/z 274.10 [M-H] ;
+
HR-ESIMS (ESI+): m/z 276.0684 calcd for C H NO S+H (276.0694).
1
4
13
3
4
.4. General procedure for propargylation: Synthesis of 14, 17 and 25. The intermediate 16
was prepared from 11 via PMB protection. Briefly, to the solution of 11 (10 mmol), and DBU
o
(
11 mmol) in DMF, the PMB-chloride (11 mmol) was added dropwise at 0 C and allowed to stir
for ten hours. The reaction mixture was concentrated under reduced pressure. Product was
directly used for next step (propargylation). The protocol for propargylation is as follows: To the
solution of aryl sulfonamide (11, 16 or 22, 6.0 mmol) and propargyl bromide (13, 18 mmol) in
acetonitrile was added K CO (18 mmol). The mixture was heated to 80°C for an hour. After the
2
3
complete consumption of the aryl sulfonamide (monitored by TLC), the reaction mixture was
cooled to room temperature. The potassium salts were filtered off and the mixture was
concentrated under reduced pressure. The residue was purified by silica gel column
chromatography using n-hexane: EtOAc as an eluent.
4
1
7
.4.1. N-(4-(1,1,1,3,3,3-Hexafluoro-2-(prop-2-yn-1-yloxy)propan-2-yl)phenyl)-N-(prop-2-yn-
1
-yl)benzenesulfonamide (14). H NMR (CDCl , 400 MHz): δ 7.71- 7.66 (m, 2H, Ar-CH), 7.59-
3
2 2
.57 (m, 3H, Ar-CH), 7.49-7.41 (m, 4H, Ar-CH), 4.49 (s, 2H, CH ), 4.25 (s, 2H, CH ), 2.57 (s,
10

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1
3
1
1
H, ≡CH), 2.20 (s, 1H, ≡CH); C NMR (CDCl 100 MHz): δ 141.74, 138.29, 133.28, 128.96,
3,
1
28.83, 127.93, 127.91, 126.88, 122.01 (d, JCF = 286 Hz, CF ), 83.5 (Ar-C-OH), 75.94 (-
3
C≡CH), 74.43 ((-C≡CH), 55.32 (-OCH
2
2 3
-), 40.75 (-NCH -); IR (CHCl ): νmax 3437, 3299, 2934,
-1
1
609, 1511, 1448, 1357, 1283, 1260, 1220, 1193, 1168, 1109, 1094, 1019 cm .
4
.4.2. N-(4-(1,1,1,3,3,3-Hexafluoro-2-((4-methoxybenzyl)oxy)propan-2-yl)phenyl)-N-(prop-2-
1
yn-1-yl)benzenesulfonamide (17). H NMR (CDCl
3
, 400 MHz): δ 7.66 (d, J = 7.5 Hz, 2H, Ar-
CH), 7.61 (d, J = 7.4 Hz, 1H, Ar-CH), 7.49 (dd, J = 14.3, 7.9 Hz, 4H, Ar-CH), 7.11 (d, J = 8.5
Hz, 4H, Ar-CH), 6.75 (d, J = 8.6 Hz, 2H, Ar-CH), 4.70 (s, 2H, O-CH
2
2
), 4.19 (s, 2H, N-CH ),
3
.74 (s, 3H, OMe).
1
4
.4.3. N-(4-Acetylphenyl)-N-(prop-2-yn-1-yl)benzenesulfonamide (25). H NMR (CDCl 400
3
,
MHz): δ 7.91 (d, J = 8.6 Hz, 2H, Ar-CH), 7.66-7.68 (m, 2H, Ar-CH), 7.58 (t, J = 5.8 Hz, 1H, Ar-
2
CH), 7.45 (t, J = 7.4 Hz, 2H, Ar-CH), 7.39 (d, J = 8.6 Hz, 2H), 4.50 (s, 2H, N-CH ), 2.60 (s, 3H,
+
-
COMe); ESI-MS: m/z 314.0 [M+H] ; HR-ESIMS (ESI+): m/z 314.08306 calcd for
+
C H NO S+H (314.08509).
17
15
3
4
.5. Procedure for PMB deprotection: Synthesis of 18. To the PMB-protected intermediate 17
was added 10% TFA in DCM and allowed to stir for an hour. The reaction mixture was
neutralized by addition of triethylamine and concentrated under reduced pressure. The residual
crude product was purified by silica gel column chromatography using a gradient mixture of
hexane and ethyl acetate.
4
.5.1. N-(4-(1,1,1,3,3,3-Hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-(prop-2-yn-1-
1
yl)benzenesulfonamide (18). H NMR (CDCl , 400 MHz): δ 7.88 (d, J = 7.5 Hz, 2H, Ar-CH),
3
7
.59 (t, J = 7.4 Hz, 1H, Ar-CH), 7.53-7.47 (m, 4H, Ar-CH), 7.27 (s, 1H, Ar-CH), 7.19 (d, J = 8.8
Hz, 1H, Ar-CH), 4.18 (s, 2H, N-CH ); IR (CHCl ): νmax 3294, 2921, 2850, 2134, 1614, 1515,
2
3
11

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-1
1
4
449, 1404, 1343, 1311, 1285, 1259, 1219, 1191, 1161, 1109, 1092, 1024 cm ; ESI-MS: m/z
+
35.90 [M-H] .
4
.6.
Procedure
for
N-benzylation:
Synthesis
of
N-(4-acetylphenyl)-N-
benzylbenzenesulfonamide (24). To the solution of the N-(4-acetylphenyl) benzenesulfonamide
18a, 1 mmol) and benzyl bromide (3 mmol) in acetonitrile was added K CO (3 mmol). The
(
2
3
mixture was heated to 80 °C for an hour. After the complete consumption of the N-(4-
acetylphenyl) benzenesulfonamide (monitored by TLC), the reaction mixture was cooled to room
temperature, and the potassium salts were filtered off. The mixture was concentrated under
reduced pressure. The residue was purified by silica gel column chromatography using hexane:
EtOAc as an eluent.
1
4
7
7
.6.1. N-(4-Acetylphenyl)-N-benzylbenzenesulfonamide (24). H NMR (CDCl3, 400 MHz): δ
.79 (d, J = 8.6 Hz, 2H, Ar-CH), 7.69-7.59 (m, 3H, Ar-CH), 7.50 (t, J = 7.5 Hz, 2H, Ar-CH),
.24-7.18 (m, 5H, Ar-CH), 7.12 (d, J = 8.6 Hz, 2H, Ar-CH), 4.77 (s, 2H, N-CH -Ar), 2.53 (s, 3H,
2
COMe); IR (CHCl ): ν 3356, 3063, 2920, 2846, 1683, 1600, 1496, 1446, 1408, 1351, 1309,
3
max
-1
+
1
292, 1266, 1218, 1167, 1091, 1057, 1027 cm ; ESI-MS: m/z 366.00 [M+H] ; HRMS (ESI+):
+
m/z 366.1154 calcd for C H NO S+H (366.1185).
2
1
19
3
4
.7. General procedure for N-trifluoromethyl methylation: Synthesis of 6, 23. To a solution
of the aryl sulfonamide (11 or 22, 1 mmol) and 2, 2, 2-trifluoroethyl trifluoromethylsulfonate
12, 2 mmol) in acetonitrile was added K CO (2 mmol). The mixture was heated to 80 °C for an
(
2
3
hour. After the complete consumption of the aryl sulfonamide (monitored by TLC), the reaction
mixture was cooled to room temperature. Potassium salts were filtered off and the mixture was
concentrated under reduced pressure. The crude product was purified by silica gel column
chromatography using hexane: EtOAc as an eluent.
12

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4
.7.1. N-(4-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-(2,2,2-trifluoroethyl)
1
benzenesulfonamide (6) [20]. H NMR (CDCl , 400 MHz): δ 7.91-7.86 (m, 2H, Ar-CH), 7.60 (t,
3
J = 7.4 Hz, 1H, Ar-CH), 7.49 (dd, J = 15.6, 8.2 Hz, 4H, Ar-CH), 7.20 (d, J = 8.9 Hz, 2H, Ar-
+
CH), 3.92-3.86 (m, 2H, N-CH ); ESI-MS: m/z 479.90 [M+H] ; HR-ESIMS: m/z 480.0329 calcd
2
+
for C H F NO S-H (480.0316).
1
7
12
9
3
4
.7.2. N-(4-Acetylphenyl)-N-(2,2,2-trifluoroethyl)benzenesulfonamide (23) [21]. White solid;
1
H NMR (CDCl3, 400 MHz): δ 7.96-7.88 (m, 2H, Ar-CH), 7.60 (dd, J = 11.4, 5.0 Hz, 3H, Ar-
CH), 7.48 (t, J = 6.9 Hz, 2H, Ar-CH), 7.22-7.13 (m, 2H, Ar-CH), 4.32-4.20 (m, 2H, N-CH2),
2
1
.65 (s, 3H, COMe); IR (CHCl ): νmax 3435, 2922, 1685, 1601, 1447, 1422, 1360, 1332, 1265,
3
-1
+
165, 1089, 1017 cm ; ESI-MS: m/z 357.90 [M+H] ; HR-ESIMS: m/z 358.0705 calcd for
+
C H NO S+H (358.0725).
14
13
3
4
.8. General procedure for click reaction: Synthesis of 20a-o and 26a-i, 26n, 26p-x. In a
round-bottom flask equipped with a magnetic stirring bar, aryl azide (19a-x, 0.70 mmol) was
added, along with N-aryl-N-(propargyl)benzenesulfonamide 18 or 25 (0.63 mmol), tert-butanol
(
0.7 mL), CuSO pentahydrate (0.03 mmol), sodium ascorbate (0.09 mmol), and H O (0.7 mL).
4
2
The reaction mixture was stirred for 48 h at room temperature and subsequently extracted with
ethyl acetate. The combined organic extracts were washed with H O, dried over anhydrous
2
2 4
Na SO , filtered, and concentrated in vacuo. The residual crude product was purified by silica
gel column chromatography using a gradient mixture of hexane and ethyl acetate.
4
1
.8.1.
N-(4-(1,1,1,3,3,3-Hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-((1-phenyl-1H-
1
,2,3-triazol-4-yl)methyl)benzenesulfonamide (20a). White solid; m.p. 69-71 °C; H NMR
(
CDCl , 400 MHz): δ 8.08 (s, 1H, Ar-CH), 7.87 (d, J = 7.4 Hz, 2H, Ar-CH), 7.76 (d, J = 7.6 Hz,
3
2
H, Ar-CH), 7.64-7.53 (m, 5H, Ar-CH), 7.48 (m, 3H, Ar-CH), 7.22 (d, J = 8.8 Hz, 2H, Ar-CH),
13

ACCEPTED MANUSCRIPT
1
3
7
1
1
1
.08 (s, 1H, OH), 4.79 (s, 2H, N-CH
2
); C NMR (CDCl
3
, 125 MHz) δ 143.97, 139.06, 138.99,
1
36.77, 133.38, 129.85, 129.45, 129.26, 129.11, 127.17, 122.98, 122.22 (d, J = 288.1 Hz),
C-F
21.33, 121.07, 119.83, 82.75 (Ar-C-OH), 60.24 (N-CH ); IR (CHCl ): ν 3436, 2920, 2851,
max
2
3
-1
633, 1515, 1466, 1409, 1346, 1259, 1218, 1190, 1161, 1106, 1022 cm ; ESI-MS: m/z 556.90
+
+
[
M+H] ; HR-ESIMS: m/z 557.1095 calcd for C H F N O S+H (557.1082).
24 18 6 4 3
4
.8.2. N-(4-(1,1,1,3,3,3-Hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-((1-(p-tolyl)-1H-1,2,3-
1
triazol-4-yl)methyl)benzenesulfonamide (20b). White solid; m.p. 74-76 °C; H NMR (CDCl
3
,
4
8
00 MHz): δ 8.07 (s, 1H, Ar-CH), 7.96-7.86 (m, 2H, Ar-CH), 7.81 (s, 1H, Ar-CH), 7.65 (d, J =
.4 Hz, 2H, Ar-CH), 7.63-7.56 (m, 3H, Ar-CH), 7.49 (dd, J = 10.5, 4.7 Hz, 2H, Ar-CH), 7.36 (d,
J = 8.1 Hz, 2H, Ar-CH), 7.29 (d, J = 2.7 Hz, 1H, Ar-CH), 4.81 (s, 2H, N-CH ), 2.46 (s, 3H, Ar-
2
13
CH ); C NMR (CDCl , 100 MHz): δ 143.83, 139.25, 139.13, 139.00, 134.58, 133.35, 130.32,
3
3
1
1
29.50, 129.25, 127.19, 123.32, 122.28 (d, JC-F = 288.1 Hz), 121.22, 120.61, 120.00, 82.75 (Ar-
C-OH), 60.36, 21.09; IR (CHCl ): νmax 3436, 2921, 2851, 1615, 1517, 1464, 1411, 1312, 1281,
218, 1190, 1161, 1108, 1020 cm ; ESI-MS: m/z 570.90 [M+H] ; HR-ESIMS: m/z 571.1228
3
-1
+
1
+
calcd for C H F N O S+H (571.1239).
2
5
20
6
4
3
4
1
.8.3. N-(4-(1,1,1,3,3,3-Hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-((1-(4-methoxyphenyl)-
1
H-1,2,3-triazol-4-yl)methyl)benzenesulfonamide (20c). White solid; m.p. 68-70 °C; H NMR
3
(CDCl , 400 MHz): δ 7.99 (s, 1H, Ar-CH), 7.89-7.84 (m, 2H, Ar-CH), 7.68-7.63 (m, 2H, Ar-
CH), 7.59-7.56 (m, 3H, Ar-CH), 7.48 (t, J = 7.6 Hz, 2H, Ar-CH), 7.41 (s, 1H, Ar-CH), 7.23 (d, J
1
3
=
8.9 Hz, 1H, Ar-CH), 7.07-7.01 (m, 2H, Ar-CH), 4.78 (s, 2H, N-CH ), 3.88 (s, 3H, OMe); C
2
NMR (CDCl , 125 MHz): δ 160.04, 143.74, 139.04, 138.96, 133.41, 130.25, 129.52, 129.29,
3
1
1
6
27.20, 123.23, 122.37, 122.25 (d, J
= 288.1 Hz), 121.46, 119.92, 114.86, 82.75 (Ar-C-OH),
C-F
0.32 (OCH ), 55.68 (OMe); IR (CHCl ): ν 3436, 2923, 1613, 1518, 1465, 1448, 1345, 1311,
max
2
3
14

ACCEPTED MANUSCRIPT
-1
+
1
258, 1218, 1190, 1162, 1106 cm ; ESI-MS: m/z 586.90 [M+H] ; HR-ESIMS: m/z 587.1183
+
calcd for C H F N O S+H (587.1188).
2
5
20
6
4
4
4
.8.4. N-(4-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-((1-(4-
(trifluoromethoxy)phenyl)-1H-1,2,3-triazol-4-yl)methyl)benzenesulfonamide (20d). White
1
solid; m.p. 67-69 °C; H NMR (CDCl
3
, 400 MHz): δ 8.07 (s, 1H, Ar-CH), 7.90-7.85 (m, 2H, Ar-
CH), 7.84-7.79 (m, 2H, Ar-CH), 7.57 (m, 3H, Ar-CH), 7.48 (t, J = 7.7 Hz, 2H, Ar-CH), 7.41 (d,
1
3
J = 9.2 Hz, 3H, Ar-CH), 7.23 (d, J = 8.8 Hz, 1H, Ar-CH), 4.79 (s, 2H, N-CH
2
); C NMR
(
CDCl , 100 MHz) δ 149.24, 144.33, 139.11, 139.07, 135.16, 133.39, 129.45, 129.26, 127.16,
3
1
1
1
1
1
6
23.07, 122.34, 122.24 (q, JC-F = 288.1 Hz), 122.18, 121.35, 120.36 (q, J
= 256.4 Hz),
C-F
19.90, 82.8 (Ar-C-OH), 60.20; IR (CHCl ): ν
3437, 2920, 2851, 1614, 1541, 1517, 1448,
max
3
-1
+
346, 1259, 1218, 1190, 1162, 1107, 1020 cm ; ESI-MS: m/z 640.90 [M+H] ; HR-ESIMS: m/z
+
41.0925 calcd for C H F N O S+H (641.0905).
2
5
18
9
4
4
4
.8.5. N-(4-(1,1,1,3,3,3-Hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-((1-(4-
(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl) methyl)benzenesulfonamide (20e). White solid;
1
m.p. 141-143 °C; H-NMR (CDCl , 500 MHz): δ 8.14 (s, 1H, Ar-CH), 7.94 (d, J = 8.5 Hz, 2H,
3
Ar-CH), 7.85 (dd, J = 13.2, 8.2 Hz, 4H, Ar-CH), 7.59 (m, 3H, Ar-CH), 7.50 (t, J = 7.7 Hz, 2H,
1
3
Ar-CH), 7.22 (d, J = 8.7 Hz, 1H, Ar-CH), 7.11 (s, 1H), 4.80 (s, 2H, N-CH ); C NMR (DMSO,
2
3
1
1
1
25MHz) δ 143.46, 140.69, 139.98, 139.74, 133.74, 129.96, 129.70, 127.72 (q, JC-F= 3.8 Hz),
3
27.13, 124.20, 121.23, 121.17, 119.43, 60.03; IR (CHCl ): νmax 3436, 2923, 2852, 1616, 1516,
-1
+
448, 1325, 1282, 1259, 1219, 1190, 1163, 1092, 1070, 1020 cm ; ESI-MS: m/z 622.80 [M-H] ;
+
HR-ESIMS: m/z 625.0948 calcd for C H F N O S+H (625.0956).
2
5
17
9
4
3
4
1
.8.6. N-(4-(1,1,1,3,3,3-Hexafluoro-2-hydroxypropan-2-yl)phenyl)-N-((1-(4-nitrophenyl)-1H-
1
,2,3-triazol-4-yl)methyl)benzenesulfonamide (20f). Yellow solid; m.p. 85-84 °C; H NMR
15

ACCEPTED MANUSCRIPT
(
CDCl
3
, 400 MHz): δ 8.48-8.42 (m, 2H, Ar-CH), 8.19 (s, 1H, Ar-CH), 8.05- 8.00 (m, 2H, Ar-
CH), 7.89-7.84 (m, 2H, Ar-CH), 7.60 (m, 3H, Ar-CH), 7.50 (t, J = 7.6 Hz, 2H, Ar-CH), 7.22 (d,
1
3
J = 8.9 Hz, 1H, Ar-CH), 6.91 (s, 1H, Ar-CH), 4.82 (s, 2H, N-CH ); C NMR (CDCl , 125 MHz)
2
3
δ 147.42, 144.91, 140.95, 139.00, 138.97, 133.52, 129.49, 129.35, 127.18, 125.65, 123.05,
1
1
3
5
22.22 (q, J
= 288.1 Hz), 121.10, 120.70, 119.87, 82.8 (Ar-C-OH), 60.12; IR (CHCl ): ν
C-F 3 max
-1
436, 2919, 1599, 1529, 1448, 1341, 1312, 1259, 1218, 1190, 1161, 1019 cm ; ESI-MS: m/z
+
+
99.90 [M-1] ; HR-ESIMS: m/z 602.0919 calcd for C H F N O S+H (602.0933).
2
4
17
6
5
5
4
.8.7. N-((1-(4-Acetylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-hexafluoro-2-
1
hydroxypropan-2-yl)phenyl)benzenesulfonamide (20g). White solid; m.p. 87-89 °C; H NMR
(
CDCl , 400 MHz): δ 8.17 (d, J = 4.5 Hz, 2H, Ar-CH), 8.14 (s, 1H, Ar-CH), 7.89 (dd, J = 13.2,
3
8
2
.1 Hz, 4H, Ar-CH), 7.63 (s, 1H, Ar-CH), 7.57 (t, J = 7.6 Hz, 3H, Ar-CH), 7.48 (t, J = 7.7 Hz,
1
3
H, Ar-CH), 7.25 (d, J = 8.8 Hz, 1H, Ar-CH), 4.80 (s, 2H, N-CH ), 2.68 (s, 3H, COMe); C
2
NMR (CDCl
3
, 125 MHz): δ 197.02, 144.42, 139.85, 139.20, 139.06, 137.03, 133.39, 130.21,
1
1
2
29.45, 129.27, 127.16, 122.90, 122.22 (d, JC-F = 285 Hz), 121.22, 120.24, 119.83, 60.13,
-1
6.75; IR (CHCl ): ν
3436, 2919, 1606, 1410, 1261, 1160, 1020 cm ; ESI-MS: m/z 598.90
max
3
+
+
[
M+H] ; HR-ESIMS: m/z 599.1182 calcd for C H F N O S+H (599.1188).
2
6
20
6
4
4
4
.8.8. N-((1-(2-Fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-hexafluoro-2-
1
hydroxypropan-2-yl)phenyl)benzenesulfonamide (20h). White solid; m.p. 124-126°C; H NMR
(
CDCl
3
, 400 MHz): δ 8.16 (d, J = 2.5 Hz, 1H, Ar-CH), 8.02- 7.93 (m, 1H, Ar-CH), 7.87 (d, J =
.6 Hz, 2H, Ar-CH), 7.61 (d, J = 8.7 Hz, 2H, Ar-CH), 7.56 (d, J = 7.3 Hz, 1H, Ar-CH), 7.52-
.43 (m, 3H, Ar-CH), 7.39-7.29 (m, 3H, Ar-CH), 7.23 (d, J = 8.7 Hz, 1H, Ar-CH), 4.80 (s, 2H,
7
7
13
1
N-CH ); C NMR (CDCl , 125 MHz) δ 153.39 (d, J = 251.2 Hz), 143.61, 139.00, 138.91,
C-F
2
3
3
4
1
33.44, 130.54 (d, J = 7.8 Hz), 129.56, 129.30, 127.21, 125.39 (d, J = 3.8 Hz), 125.06 (d,
C-F
C-F
16

ACCEPTED MANUSCRIPT
2
3
1
J
C-F = 10.2 Hz), 124.94, 124.36 (d, JC-F = 8.2 Hz), 123.32, 122.24 (q, JC-F = 288.3 Hz),
2
1
1
19.93, 117.12 (d, J = 19.9 Hz), 82.8 (Ar-C-OH), 60.15; IR (CHCl ): ν
3436, 2919, 1613,
max
C-F
3
-1
513, 1474, 1346, 1312, 1282, 1259, 1218, 1190, 1163, 1107, 1011 cm ; ESI-MS: m/z 572.90
+
+
[
M-H] ; HR-ESIMS: m/z 575.1035 calcd for C H F N O S+H (575.0988).
24 17 7 4 3
4
.8.9. N-((1-(4-Fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-hexafluoro-2-
1
hydroxypropan-2-yl)phenyl)benzenesulfonamide (20i). White solid; m.p. 94-96 °C; H NMR
3
(CDCl , 400 MHz): δ 8.03 (s, 1H, Ar-CH), 7.90-7.84 (m, 2H, Ar-CH), 7.78-7.70 (m, 2H, Ar-
CH), 7.57 (t, J = 7.5 Hz, 3H, Ar-CH), 7.48 (t, J = 7.7 Hz, 2H, Ar-CH), 7.26-7.22 (m, 5H, Ar-
1
3
1
CH), 4.79 (s, 2H, N-CH ); C NMR (CDCl , 125 MHz): δ 162.61 (d, J = 249.7 Hz), 144.11,
2
3
C-F
4
1
39.01, 138.93, 133.46, 133.11 (d, J = 3.1 Hz), 129.52, 129.31, 127.19, 123.23, 122.73 (d,
C-F
3
1
2
JC-F = 8.7 Hz), 122.24 (d, J = 288.0 Hz), 121.42, 119.92, 116.87 (d, J = 23.3 Hz), 60.27;
C-F
C-F
-1
IR (CHCl ): ν
3443, 2928, 1615, 1516, 1448, 1282, 1219, 1190, 1162, 1092, 1022 cm ; ESI-
max
3
+
+
17 7 4 3
MS: m/z 572.90 [M+H] ; HR-ESIMS: m/z 575.1049 calcd for C24H F N O S+H (575.0988).
4
2
.8.10. N-((1-(2,6-Difluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-hexafluoro-
1
-hydroxypropan-2-yl)phenyl)benzenesulfonamide (20j). White solid; m.p. 71-73 °C; H NMR
(
CDCl , 400 MHz): δ 7.93 (s, 1H, Ar-CH), 7.90-7.84 (m, 2H, Ar-CH), 7.62- 7.43 (m, 7H, Ar-
3
1
3
CH), 7.24 (d, J = 8.9 Hz, 2H, Ar-CH), 7.16 (t, J = 8.2 Hz, 1H, Ar-CH), 4.82 (s, 2H, N-CH ); C
2
1
NMR (CDCl
3
, 125 MHz) δ 156.82 (d, JC-F= 256.9 Hz), 143.23, 138.98, 138.84, 133.47, 131.60
3
2
(
d, JC-F= 9.7 Hz), 129.56, 129.31, 127.21, 125.58, 123.39, 120.01, 112.65 (d, JC-F = 19.6 Hz),
6
1
0.19; IR (CHCl ): ν
3443, 3256, 2927, 1610, 1516, 1481, 1312, 1284, 1259, 1218, 1204,
max
3
-1
+
190, 1163, 1106, 1092, 1013 cm ; ESI-MS: m/z 592.90 [M+H] ; HR-ESIMS: m/z 593.0886
+
calcd for C H F N O S+H (593.0894).
2
4
16
8
4
3
17

ACCEPTED MANUSCRIPT
4
2
.8.11. N-((1-(2,4-Difluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-hexafluoro-
1
-hydroxypropan-2-yl)phenyl)benzenesulfonamide (20k). White solid; m.p. 65-67 °C; H NMR
(
400 MHz, CDCl ): δ 8.12 (d, J = 2.0 Hz, 1H, Ar-CH), 7.93-7.90 (m, 4H, Ar-CH), 7.57 (dd, J =
3
1
2
2.1, 8.0 Hz, 3H, Ar-CH), 7.47 (t, J = 7.6 Hz, 2H, Ar-CH), 7.27 (s, 1H, Ar-CH), 7.13-7.05 (m,
1
3
1
H, Ar-CH), 4.79 (s, 2H, N-CH ); C NMR (CDCl 125 MHz): δ 162.64 (d, J = 253.2 Hz),
2
3,
C-F
1
1
53.88 (d, J = 254.0 Hz), 143.75, 139.07, 139.01, 133.41, 129.49, 129.28, 127.19, 126.34 (d,
C-F
3
4
1
2
JC-F = 10.0 Hz), 124.32 (d, J = 7.2 Hz), 123.10, 122.23 (q, J = 288.1 Hz), 121.68 (d, J
C-F
C-F
C-F
2
2
=
10.7 Hz), 119.86, 112.75 (d, JC-F = 22.7 Hz), 105.49 (d, JC-F = 26.7 Hz), 82.79 (Ar-C-OH),
+
+
6
0.08; ESI-MS: m/z 592.90 [M+H] ; HR-ESIMS: m/z 593.0874 calcd for C24
16 8 4 3
H F N O S+H
(593.0894).
4
.8.12. N-((1-(3,4-Dimethylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-
hexafluoro-2-hydroxypropan-2-yl)phenyl)benzenesulfonamide (20l). White solid; m.p. 71-73
1
°
C; H NMR (CDCl
3
, 500 MHz): δ 8.03 (s, 1H, Ar-CH), 7.90-7.82 (m, 2H, Ar-CH), 7.59 (m,
3
H, Ar-CH), 7.54 (s, 1H, Ar-CH), 7.50 (t, J = 7.8 Hz, 2H, Ar-CH), 7.45 (d, J = 8.1 Hz, 1H, Ar-
CH), 7.29 (d, J = 5.3 Hz, 1H, Ar-CH), 7.21 (d, J = 8.8 Hz, 1H, Ar-CH), 6.89 (s, 1H, Ar-CH),
1
3
4
1
1
.78 (s, 2H, N-CH ), 2.37 (s, 3H, Ar-CH ), 2.34 (s, 3H, Ar-CH ); C NMR (CDCl , 125 MHz) δ
2 3 3 3
43.73, 139.06, 139.02, 138.51, 137.93, 134.73, 133.39, 130.72, 129.50, 129.27, 127.20, 123.19,
1
22.26 (q, JC-F= 288.1 Hz), 121.83, 121.32, 119.91, 118.01, 60.34, 19.93, 19.52; IR (CHCl3):
-1
+
νmax 3436, 2921, 2850, 1615, 1190, 1161, 1021 cm ; ESI-MS: m/z 584.90 [M+H] ; HR-ESIMS:
+
m/z 585.1389 calcd for C26
H
22
F
6
N
4
O
3
S+H (585.1395).
4
.8.13. N-((1-(3-Fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-hexafluoro-2-
1
hydroxypropan-2-yl)phenyl)benzenesulfonamide (20m). White solid; m.p. 69-71 °C; H NMR
CDCl , 400 MHz): δ 8.08 (s, 1H, Ar-CH), 7.91-7.83 (m, 2H, Ar-CH), 7.62- 7.46 (m, 8H, Ar-
(
3
18

ACCEPTED MANUSCRIPT
13
CH), 7.25-7.14 (m, 3H, Ar-CH), 4.79 (s, 2H, N-CH
2
); C NMR (CDCl
3
,125 MHz) δ 162.57 (d,
1
3
3
JC-F = 248.9 Hz), 143.75, 138.66, 138.53, 137.40 (d, JC-F = 10.0 Hz), 132.90, 130.86 (d, J
=
C-F
1
8
.9 Hz), 128.94, 128.78, 126.68, 122.46, 121.74 (d, J = 290.5 Hz), 120.81, 119.36, 115.52 (d,
C-F
2
4
2
JC-F = 21.1 Hz), 115.50 (d, J = 3.2 Hz), 107.94 (d, J = 26.2 Hz), 59.67; IR (CHCl ): ν
max
C-F
C-F
3
-1
3
436, 2921, 1612, 1515, 1463, 1406, 1345, 1311, 1282, 1259, 1218, 1190, 1162 cm ; ESI-MS:
+
+
m/z 572.90 [M+H] ; HR-ESIMS: m/z 573.0849 calcd for C H F N O S-H (573.0831).
2
4
17
7
4
3
4
.8.14.
Ethyl 4-(4-((N-(4-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)phenyl) phenyl
1
sulfonamido) methyl)-1H-1,2,3-triazol-1-yl)benzoate (20n). White solid; m.p. 109-111 °C; H
NMR (CDCl , 500 MHz): δ 8.24 (d, J = 8.6 Hz, 2H, Ar-CH), 8.17 (s, 1H, Ar-CH), 7.88 (d, J =
3
8
1
.4 Hz, 4H, Ar-CH), 7.58 (t, J = 7.1 Hz, 3H, Ar-CH), 7.49 (t, J = 7.7 Hz, 2H, Ar-CH), 7.42 (s,
H, Ar-CH), 7.24 (d, J = 8.8 Hz, 2H, Ar-CH), 4.80 (s, 2H, N-CH ), 4.44 (q, J = 7.1, 10 Hz, 2H, -
2
1
3
COOCH CH ), 1.44 (t, J = 7.1 Hz, 3H, -COOCH CH ); C NMR (CDCl , 125 MHz) δ 165.46,
2
3
2
3
3
1
1
44.39, 139.79, 139.02, 139.00, 133.45, 131.40, 130.87, 129.51, 129.31, 127.19, 122.22 (q, JC-F
288.0 Hz), 123.13, 121.10, 120.04, 119.90, 82.80 (m), 61.59, 60.21, 14.32; IR (CHCl
=
3
): νmax
-1
3
436, 2922, 1715, 1610, 1516, 1447, 1280, 1218, 1162, 1107, 1021 cm ; ESI-MS: m/z 628.90
+
+
[
M+H] ; HR-ESIMS: m/z 629.1282 calcd for C H F N O S+H (629.1293).
27 22 6 4 5
4
.8.15.
N-((1-(2,5-Dimethylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-N-(4-(1,1,1,3,3,3-
hexafluoro -2-hydroxypropan-2-yl)phenyl)benzenesulfonamide (20o). White solid; m.p. 74-76
1
°
C; H NMR (CDCl
3
, 500 MHz): δ 7.86 (d, J = 7.4 Hz, 2H, Ar-CH), 7.82 (s, 1H, Ar-CH), 7.59
(m, 3H, Ar-CH), 7.49 (t, J = 7.6 Hz, 2H, Ar-CH), 7.26-7.24 (m, 1H, Ar-CH), 7.24-7.18 (m, 3H,
Ar-CH), 7.11 (s, 1H, Ar-CH), 4.80 (s, 2H, N-CH ), 2.39 (s, 3H, Ar-CH ), 2.19 (s, 3H, Ar-CH );
2
3
3
1
3
C NMR (126 MHz, CDCl ): δ 143.05, 139.05, 138.98, 136.98, 136.01, 133.40, 131.37, 130.82,
3
1
1
30.33, 129.52, 129.29, 127.20, 126.50, 124.54, 123.26, 122.27 (q, J = 288.2 Hz), 119.91,
C-F
19

ACCEPTED MANUSCRIPT
8
1
5
3
2.76 (Ar-C-OH), 60.33, 20.73, 17.45; IR (CHCl ): νmax 3436, 2924, 1614, 1515, 1465, 1448,
-1
+
406, 1346, 1312, 1189, 1162, 1045, 1016 cm ; ESI-MS: m/z 584.90 [M+H] ; HR-ESIMS: m/z
+
85.1383 calcd for C H F N O S+H (585.1395).
2
6
22
6
4
3
4
.8.16. N-(4-Acetylphenyl)-N-((1-phenyl-1H-1,2,3-triazol-4-yl)methyl)benzenesulfonamide
1
(
26a): White solid; m.p. 189-191 °C; H NMR (CDCl3, 400 MHz): δ 8.02 (s, 1H, Ar-CH), 7.86
(d, J = 8.4 Hz, 2H, Ar-CH), 7.67 (d, J = 8.0 Hz, 2H, Ar-CH), 7.62 (d, J = 8.4 Hz, 3H, Ar-CH),
7
.49-7.47 (m, 5H, Ar-CH), 7.29 (d, J = 8.5 Hz, 1H, Ar-CH), 5.03 (s, 2H, N-CH
2
), 2.56 (s, 3H,
13
COCH3); C NMR (CDCl 125 MHz): δ 195.14, 141.32, 135.70, 134.86, 134.01, 131.39,
3,
1
2
27.86, 127.30, 127.23, 127.02, 125.82, 125.59, 118.5, 44.42, 24.71; IR (CHCl ): νmax 3400,
3
-1
915, 1681, 1599, 1502, 1445, 1360, 1265, 1238, 1207, 1161, 1091, 1061, 1041 cm ; ESI-MS:
+
+
m/z 433.00 [M+H] ; HR-ESIMS: m/z 433.1333 calcd for C H N O S+H (433.1356).
2
3
20
4
3
4
.8.17. N-(4-Acetylphenyl)-N-((1-(p-tolyl)-1H-1,2,3-triazol-4-yl)methyl)benzenesulfonamide
1
(
26b). White solid; m.p. 179-181 °C; H NMR (CDCl 400 MHz): 7.98 (s, 1H, Ar-CH), 7.86 (d,
3
,
J = 8 Hz, 2H, Ar-CH), 7.62 (d, J = 12 Hz, 3H, Ar-CH), 7.54 (d, J = 7.6 Hz, 2H, Ar-CH), 7.48
(
m, 2H, Ar-CH), 7.30 (dd, J = 4, 8 Hz, 4H, Ar-CH), 5.02 (s, 2H, N-CH ), 2.56 (s, 3H, COCH ),
2 3
1
3
2
1
.42 (s, 3H, Ar-Me); C NMR (CDCl 100 MHz): δ 196.97, 143.35, 139.08, 137.86, 135.97,
3,
34.57, 133.26, 130.25, 129.20, 129.12, 127.76, 127.53, 120.36, 46.43, 26.57, 21.07; IR
-1
(
3
CHCl ): νmax 3435, 2970, 1739, 1680, 1365, 1232, 1216, 1161, 1089, 1042 cm ; ESI-MS: m/z
+
+
4
22 4 4
47.00 [M+H] ; HR-ESIMS: m/z 447.14799 calcd for C25H N O S+H (447.15126).
4
.8.18. N-(4-Acetylphenyl)-N-((1-(4-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl) benzene
1
sulfonamide (26c). White solid; m.p. 179-181 °C; H NMR (CDCl 400 MHz): 7.94 (s, 1H, Ar-
3
,
CH), 7.86 (d, J = 8Hz, 2H, Ar-CH), 7.55 (m, 7H, Ar-CH), 7.29 (d, J = 8 Hz, 2H, Ar-CH), 7.00
1
3
(
d, J = 8 Hz, 2H, Ar-CH), 5.02 (s, 2H, N-CH
2
), 3.86 (s, 3H, OMe), 2.56 (s, 3H, COCH
3
);
C
20

ACCEPTED MANUSCRIPT
NMR (CDCl3, 125 MHz): δ 197.11, 159.93, 143.30, 137.68, 135.93, 133.37, 129.23, 129.16,
1
1
1
27.77, 127.53, 122.09, 114.79, 55.66, 46.41, 26.65; IR (CHCl ): ν
3400, 3114, 2920, 2850,
max
3
679, 1600, 1518, 1444, 1409, 1384, 1353, 1304, 1254, 1229, 1190, 1163, 1111, 1092, 1046,
-1
+
+
022 cm ; ESI-MS: m/z 463.10 [M+H] ; HR-ESIMS: m/z 463.1433 calcd for C H N O S+H
2
4
22
4
4
(463.1461).
4
.8.19. N-(4-Acetylphenyl)-N-((1-(4-(trifluoromethoxy)phenyl)-1H-1,2,3-triazol-4-
1
yl)methyl)benzenesulfonamide (26d). White solid; m.p. 162-164 °C; H NMR (CDCl3, 400
MHz): 8.03 (s, 1H, Ar-CH), 7.87 (d, J = 12 Hz, 2H, Ar-CH), 7.74 (d, J = 8 Hz, 2H, Ar-CH),
7
.62 (m, 3H, Ar-CH), 7.50 (d, J = 8 Hz, 2H, Ar-CH), 7.38 (d, J = 8 Hz, 2H, Ar-CH), 7.28 (d, J
13
=
8 Hz, 2H, Ar-CH), 5.03 (s, 2H, N-CH ), 2.56 (s, 3H, COCH ); C NMR (DMSO-d , 100
2 3 6
MHz): δ 197.04, 147.83, 143.53, 142.74, 137.48, 135.53, 135.20, 133.50, 129.43, 128.87,
1
1
27.93, 127.26, 122.54, 122.47, 121.91, 45.46, 26.66; IR (CHCl ): ν 3436, 3127, 3065, 2923,
3 max
-1
+
681, 1660, 1517, 1354, 1266, 1165, 1092, 1042 cm ; ESI-MS: m/z 517.00 [M+H] ; HR-
+
ESIMS: m/z 517.1149 calcd for C24
H
19
F
3
N
4
O
4
S+H (517.1179).
4
.8.20. N-(4-Acetylphenyl)-N-((1-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-
1
yl)methyl)benzene sulfonamide (26e). White solid; m.p. 194-196 °C; H NMR (CDCl 400
3
,
MHz): δ 8.11 (s, 1H, Ar-CH), 7.91-7.83 (m, 4H, Ar-CH), 7.80 (d, J = 8.5 Hz, 2H, Ar-CH), 7.62
(d, J = 6.7 Hz, 3H, Ar-CH), 7.50 (t, J = 7.7 Hz, 2H, Ar-CH), 7.28 (d, J = 8.7 Hz, 2H, Ar-CH),
1
3
5
1
1
.04 (s, 2H, N-CH
2
), 2.56 (s, 3H, COCH
3
3
); C NMR (CDCl , 125 MHz) δ 197.03, 144.74,
2
43.24, 139.16, 137.59, 136.05, 133.43, 130.91 (q, JC-F = 32.2 Hz), 129.30, 129.22, 127.75,
3
1
27.53, 127.16 (q, J = 3.6 Hz), 123.48 (q, J = 270.1 Hz), 121.24, 120.41, 46.35, 26.65; IR
C-F
C-F
(
CHCl ): ν 3435, 3124, 3085, 3003, 2922, 1683, 1616, 1600, 1352, 1325, 1266, 1163, 1129,
3 max
21

ACCEPTED MANUSCRIPT
-1
+
1
093, 1042 cm ; ESI-MS: m/z 501.00 [M+H] ; HR-ESIMS: m/z 501.1199 calcd for
+
C H F N O S+H (501.1230).
24
19
3
4
3
4
.8.21. N-(4-Acetylphenyl)-N-((1-(4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)benzene
1
sulfonamide (26f). Yellow solid; m.p. 169-171 °C; H NMR (CDCl 400 MHz): 8.42 (m, 2H,
3
,
Ar-CH), 8.18 (s, 1H, Ar-CH), 7.94 (d, J = 9.1Hz, 2H, Ar-CH), 7.88 (d, J = 8.6 Hz, 2H, Ar-CH),
7
3
1
.63 (d, 3H, Ar-CH), 7.50 (m, 2H, Ar-CH), 7.28 (d, 2H, Ar-CH), 5.04 (s, 2H, N-CH
2
), 2.56 (s,
, 125 MHz): δ 197.57, 147.15, 144.58, 143.21, 141.04,
37.85, 136.03, 134.05, 129.96, 129.42, 128.45, 127.77, 126.01, 123.18, 120.95, 45.92, 27.19;
1
3
3 6
H, COCH ); C NMR (DMSO-d
IR (CHCl ): ν
3848, 3744, 3399, 2919, 2850, 1682, 1597, 1523, 1384, 1344, 1266, 1162,
max
3
-1
+
1
092, 1041 cm ; ESI-MS: m/z 478.00 [M+H] ; HR-ESIMS: m/z 478.11682 calcd for
+
C H N O S+H (478.12069).
23
19
5
5
4
.8.22. N-(4-acetylphenyl)-N-((1-(4-acetylphenyl)-1H-1,2,3-triazol-4-yl)methyl)benzene
1
sulfonamide (26g). White solid; m.p. 188-190 °C; H NMR (CDCl 400 MHz) δ 8.15 (s, 1H,
3
,
Ar-CH), 8.11 (d, J = 8.5 Hz, 2H, Ar-CH), 7.85 (dd, J = 16.3, 8.5 Hz, 4H, Ar-CH), 7.66-7.59 (m,
H, Ar-CH), 7.50 (t, J = 7.7 Hz, 2H, Ar-CH), 7.28 (d, J = 3.0 Hz, 2H, Ar-CH), 5.04 (s, 2H, N-
3
1
3
CH ), 2.65 (s, 3H, COCH ), 2.56 (s, 3H, COCH ); C NMR (CDCl 125 MHz) δ 197.05,
2
3
3
3,
1
1
3
1
4
96.59, 144.65, 143.25, 139.80, 137.60, 136.97, 136.03, 133.42, 130.12, 129.30, 129.22, 127.76,
27.53, 121.23, 120.03, 46.35, 26.74, 26.66; IR (CHCl ): νmax 3849, 3812, 3741, 3685, 3443,
129, 3061, 3003, 2920, 1681, 1603, 1554, 1516, 1446, 1411, 1354, 1308, 1292, 1266, 1236,
3
-1
+
215, 1166, 1112, 1091, 1040, 1018 cm ; ESI-MS: m/z 475.00 [M+H] ; HR-ESIMS: m/z
+
75.14220 calcd for C H N O S+H (475.14618).
2
5
22
4
4
4
.8.23. N-(4-Acetylphenyl)-N-((1-(2-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)
benzene
1
sulfonamide (26h). White solid; m.p. 144-146 °C; H NMR (CDCl3, 400 MHz): 8.07 (s, 1H, Ar-
22

ACCEPTED MANUSCRIPT
CH), 7.87 (t, J = 8 Hz, 3H, Ar-CH), 7.61 (t, J = 8 Hz, 3H, Ar-CH), 7.46 (m, 4H, Ar-CH), 7.30 (d,
13
J = 8 Hz, 3H, Ar-CH), 5.05 (s, 2H, N-CH ), 2.56 (s, 3H, COCH ); C NMR (CDCl 125 MHz) δ
2
3
3,
1
2
1
7
97.13, 153.27 (d, J = 251.1 Hz), 143.53, 143.25, 137.68, 136.01, 133.31, 130.42 (d, J
=
C-F
C-F
3
2
.9 Hz), 129.24, 129.15, 127.96, 127.57, 125.28 (d, J = 3.6 Hz), 124.76, 124.54 (d, J = 8.6
C-F
C-F
Hz), 117.17, 117.01, 46.24, 26.67; IR (CHCl ): ν 3437, 3067, 2922, 2852, 1683, 1600, 1509,
3
max
-1
1
446, 1409, 1354, 1310, 1266, 1245, 1166, 1111, 1091, 1040, 1015 cm ; ESI-MS: m/z 451.00
+
+
[
M+H] ; HR-ESIMS: m/z 451.12285 calcd for C H FN O S+H (451.12619).
23 19 4 3
4
.8.24. N-(4-Acetylphenyl)-N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)benzene
1
sulfonamide (26i). White solid; m.p. 186-188 °C; H NMR (CDCl 400 MHz): 7.99 (s, 1H, Ar-
3
,
CH), 7.87 (d, J = 8.7 Hz, 2H, Ar-CH), 7.65 (m, 5H, Ar-CH), 7.49 (m, 2H, Ar-CH), 7.29 (m, 2H,
1
3
Ar-CH), 7.21 (m, 2H, Ar-CH), 5.02 (s, 2H, N-CH ), 2.56 (s, 3H, COCH ); C NMR (CDCl 125
2
3
3,
1
MHz) δ 197.16, 162.50 (d, J = 249.4 Hz), 144.25, 143.26, 137.63, 135.98, 133.39, 133.07 (d,
C-F
4
3
2
J
C-F= 3.1 Hz), 129.27, 129.20, 127.74, 127.53, 122.46 (d, JC-F = 8.6 Hz), 121.50, 116.79 (d, JC-
F
= 23.2 Hz), 46.36, 26.68; IR (CHCl ): νmax 3434, 3120, 2921, 1683, 1599, 1514, 1447, 1409,
3
-1
+
1
356, 1309, 1265, 1226, 1165, 1091, 1043 cm ; ESI-MS: m/z 451.00 [M+H] ; HR-ESIMS: m/z
+
4
51.12367 calcd for C H FN O S+H (451.12619).
2
3
19
4
3
4
.8.25. Methyl-4-(4-((N-(4-acetylphenyl)phenylsulfonamido)methyl)-1H-1,2,3-triazol-1-
1
yl)benzoate (26n). White solid; m.p. 191-193 °C; H NMR (CDCl3, 400 MHz): 8.20 (d, J =
8
2
.7Hz, 2H, Ar-CH), 8.11 (s, 1H, Ar-CH), 7.87 (d, J = 8.6 Hz, 2H, Ar-CH), 7.80 (d, J = 8.8 Hz,
H, Ar-CH), 7.62 (m, 3H, Ar-CH), 7.49 (m, 2H, Ar-CH), 7.29 (d, J = 8.7 Hz, 2H, Ar-CH), 5.04
1
3
(
s, 2H, N-CH ), 3.96 (s, 3H, -COOMe), 2.56 (s, 3H, COCH ); C NMR (CDCl 125 MHz) δ
2 3 3,
1
1
97.04, 165.83, 143.24, 139.81, 137.62, 136.02, 133.40, 131.36, 130.38, 129.28, 129.21, 127.76,
27.53, 121.26, 119.87, 52.45, 46.34, 26.65; IR (CHCl ): νmax 3789, 3435, 3126, 3059, 2951,
3
23

ACCEPTED MANUSCRIPT
2
1
850, 1720, 1681, 1607, 1518, 1445, 1411, 1354, 1310, 1282, 1235, 1166, 1111, 1091, 1040,
-
1
+
+
019 cm ; ESI-MS: m/z 491.00 [M+H] ; HR-ESIMS: m/z 491.13821 calcd for C H N O S+H
2
5
22
4
5
(491.14109).
4
.8.26.
N-(4-Acetylphenyl)-N-((1-(2,6-dimethylphenyl)-1H-1,2,3-triazol-4-yl)methyl)
1
benzenesulfonamide (26p). White solid; m.p. 141-143 °C; H NMR (CDCl3, 400 MHz): 7.84 (d,
J = 8.6 Hz, 2H, Ar-CH), 7.64 (m, 3H, Ar-CH), 7.55 (s, 1H, Ar-CH), 7.51 (t, J = 7.4 Hz, 2H, Ar-
CH), 7.30 (d, J = 7.6 Hz, 1H, Ar-CH), 7.22 (d, J = 8.6 Hz, 2H, Ar-CH), 7.13 (d, J = 6.6 Hz, 2H,
1
3
Ar-CH), 5.08 (s, 2H, N-CH ), 2.56 (s, 3H, COCH ), 1.80 (s, 6H, 2 x Ar-CH ); C NMR (CDCl
3,
2
3
3
1
1
25 MHz): δ 196.08, 142.16, 141.94, 136.50, 134.99, 134.69, 134.29, 132.46, 129.25, 128.28,
28.18, 127.54, 127.16, 126.74, 124.22, 45.40, 25.75, 16.17; IR (CHCl ): ν 3436, 3140, 3066,
3
max
-
3
004, 2924, 1683, 1600, 1573, 1504, 1487, 1446, 1354, 1266, 1237, 1166, 1092, 1070, 1042 cm
1
+
+
;
ESI-MS: m/z 461.10 [M+H] ; HR-ESIMS: m/z 461.1648 calcd for C H N O S+H
25 24 4 3
(461.1669).
4
.8.27. N-(4-Acetylphenyl)-N-((1-(4-hydroxy-3-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)
1
benzene sulfonamide (26q). Yellow solid; m.p. 163-165 °C; H NMR (CDCl 400 MHz): 10.64
3
,
(
s, 1H, OH), 8.41 (d, J = 4 Hz, 1H, Ar-CH), 8.05 (s, 1H, Ar-CH), 7.99 (dd, J = 4, 8 Hz, 1H, Ar-
CH), 7.88 (d, J = 12 Hz, 2H, Ar-CH), 7.63 (m, 3H, Ar-CH), 7.50 (m, 2H, Ar-CH), 7.35 (d, J =
Hz, 1H, Ar-CH), 7.28 (d, J = 8 Hz, 2H, Ar-CH), 5.03 (s, 2H, N-CH ), 2.56 (s, 3H, COCH );
C NMR (CDCl3, 125 MHz) δ 197.06, 154.93, 143.24, 137.59, 136.06, 133.53, 129.63, 129.32,
8
2
3
1
3
1
29.24, 127.73, 127.54, 121.85, 121.32, 116.65, 46.36, 26.67; IR (CHCl ): ν
3306, 2925,
3
max
-
2
346, 1731, 1682, 1634, 1600, 1545, 1503, 1423, 1447, 1354, 1264, 1166, 1091, 1076, 1041 cm
1
+
+
;
ESI-MS: m/z 494.00 [M+H] ; HR-ESIMS: m/z 494.1116 calcd for C H N O S+H
25 22 4 5
(494.1156).
24

ACCEPTED MANUSCRIPT
4
1
.8.28. N-(4-Acetylphenyl)-N-((1-(4-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)phenyl)-
1
H-1,2,3-triazol-4-yl)methyl)benzenesulfonamide (26r). White solid; m.p. 151-153 °C; H
NMR (CD OD, 400 MHz): δ 8.40 (s, 1H, Ar-CH), 7.83-7.79 (m, 6H, Ar-CH), 7.58 (m, 3H, Ar-
3
CH), 7.50-7.44 (m, 2H, Ar-CH), 7.21 (d, J = 8.6 Hz, 2H, Ar-CH), 4.98 (s, 2H, N-CH ), 2.45 (s,
2
1
3
3
1
3
1
H, COCH ); C NMR (CD OD, 125 MHz): δ 199.40, 144.66, 139.28, 139.12, 137.41, 134.60,
3 3
33.32, 130.39, 130.25, 129.95, 129.58, 128.83, 123.62, 121.17, 46.76, 26.74; IR (CHCl ): νmax
3
891, 3880, 3853, 3821, 3807, 3801, 3779, 3744, 3702, 3400, 2920, 2851, 2356, 1683, 1598,
-1
517, 1447, 1432, 1410, 1353, 1272, 1220, 1195, 1166, 1117, 1058 cm ; ESI-MS: m/z 598.90
+
+
[
20 6 4 4
M+H] ; HR-ESIMS: m/z 599.11742 calcd for C26H F N O S+H (599.12095).
4
.8.29. N-(4-Acetylphenyl)-N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)benzene
1
sulfonamide (26s). White solid; m.p. 203-205 °C; H NMR (CDCl 400 MHz): 8.02 (s, 1H, Ar-
3
,
CH), 7.86 (d, J = 8.6 Hz, 2H, Ar-CH), 7.64-7.61 (m, 5H, Ar-CH), 7.49-7.46 (m, 4H, Ar-CH),
1
3
7
1
1
3
1
.28 (d, J = 8.6 Hz, 2H, Ar-CH), 5.02 (s, 2H, N-CH
00 MHz): δ 196.98, 143.29, 137.74, 136.03, 135.30, 134.76, 133.34, 129.97, 129.26, 129.17,
27.72, 127.52, 121.62, 46.39, 26.60; IR (CHCl ): ν 3914, 3889, 3861, 3835, 3799, 3763,
2 3
), 2.56 (s, 3H, COCH ); C NMR (CDCl3,
3
max
748, 3742, 3707, 3699, 3673, 3667, 3645, 3626, 3584, 3418, 3121, 2919, 1679, 1599, 1552,
-1
+
499, 1355, 1265, 1235, 1213, 1162, 1092, 1042, 1013 cm ; ESI-MS: m/z 467.00 [M+H] ; HR-
+
ESIMS: m/z 467.09372 calcd for C H N O S+H (467.09664).
2
5
22
4
5
4
.8.30. N-(4-Acetylphenyl)-N-((1-(3-acetylphenyl)-1H-1,2,3-triazol-4-yl)methyl)benzene
1
sulfonamide (26t). White solid; m.p. 149-151 °C; H NMR (CDCl 400 MHz): 8.24 (s, 1H, Ar-
3
,
CH), 8.11 (s, 1H, Ar-CH), 8.01 (d, J = 4 Hz, 1H, Ar-CH), 7.94 (d, J = 8 Hz, 1H, Ar-CH), 7.87
(d, J = 8 Hz, 2H, Ar-CH), 7.64-7.62 (m, 4H, Ar-CH), 7.50 (t, J = 8 Hz, 2H, Ar-CH), 7.29 (d, J
13
=
8 Hz, 2H, Ar-CH), 5.04 (s, 2H, N-CH ), 2.67 (s, 3H, COCH ), 2.56 (s, 3H, COCH ); C NMR
2 3 3
25

ACCEPTED MANUSCRIPT
(
DMSO-d
6
, 100 MHz) δ 197.09, 197.04, 143.50, 142.77, 138.19, 136.64, 135.54, 135.21, 133.50,
1
30.42, 129.44, 128.88, 128.18, 127.93, 127.27, 124.26, 122.51, 119.24, 45.49, 26.92, 26.67; IR
(
CHCl ): ν 3443, 3145, 3068, 3011, 2924, 2442, 1685, 1600, 1502, 1490, 1475, 1447, 1427,
3 max
-1
+
1
356, 1310, 1291, 1263, 1167, 1091, 1041, 1017 cm ; ESI-MS: m/z 475.10 [M+H] ; HR-
+
ESIMS: m/z 475.14245 calcd for C H N O S+H (475.14618).
2
5
22
4
4
4
.8.31. N-(4-Acetylphenyl)-N-((1-(3-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)benzene
1
sulfonamide (26u). Yellow solid; m.p. 159-161°C; H NMR (CDCl3, 400 MHz): δ 8.58 (d, J =
1
1
.8 Hz, 1H, Ar-CH), 8.31 (d, J = 8.2 Hz, 1H, Ar-CH), 8.17 (s, 1H, Ar-CH), 8.11 (d, J = 7.4 Hz,
H, Ar-CH), 7.88 (d, J = 8.5 Hz, 2H, Ar-CH), 7.75 (t, J = 8.2 Hz, 1H, Ar-CH), 7.64 (d, J = 8.9
Hz, 3H, Ar-CH), 7.51 (t, J = 7.7 Hz, 2H, Ar-CH), 7.29 (d, J = 8.7 Hz, 2H, Ar-CH), 5.05 (s, 2H,
1
3
N-CH ), 2.57 (s, 3H, COCH ); C NMR (CDCl 125 MHz): δ 197.09, 148.90, 143.21, 137.54,
2
3
3,
1
2
1
37.49 136.07, 133.48, 131.04, 129.33, 129.26, 127.78, 127.54, 125.86, 123.38, 115.33, 46.32,
6.68; IR(CHCl ): νmax 3790, 3439, 3099, 2924, 1731, 1682, 1600, 1534, 1447, 1409, 1350,
3
-
1
+
266, 1165, 1091, 1040 cm ; ESI-MS: m/z 478.00 [M+H] ; HR-ESIMS: m/z 478.11737 calcd
+
for C H N O S+H (478.12069).
2
5
22
4
4
4
.8.32. N-(4-Acetylphenyl)-N-((1-(o-tolyl)-1H-1,2,3-triazol-4-yl)methyl)benzenesulfonamide
1
(
26v). White solid; m.p. 170-172 °C; H NMR (CDCl 400 MHz): δ 7.86 (d, J = 8.7 Hz, 2H, Ar-
3
,
CH), 7.73 (s, 1H, Ar-CH), 7.62 (dd, J = 10.6, 4.7 Hz, 3H, Ar-CH), 7.50 (dd, J = 8.6, 7.0 Hz, 2H,
Ar-CH), 7.40 (dd, J = 10.6, 4.3 Hz, 1H, Ar-CH), 7.36- 7.28 (m, 3H, Ar-CH), 7.25 (m, 2H, Ar-
1
3
CH), 5.06 (s, 2H, N-CH ), 2.56 (s, 3H, COCH ), 2.05 (s, 3H, Ar-Me); C NMR (CDCl 100
2
3
3,
MHz): δ 196.94, 143.34, 137.7, 136.2, 135.98, 133.50, 133.28, 131.48, 130.02, 129.16, 129.14,
1
3
27.89, 127.57, 126.88, 125.87, 124.87, 99.99, 46.39, 26.58, 17.63; IR (CHCl ): ν 3435, 3141,
3 max
-
066, 2924, 1683, 1600, 1503, 1446, 1409, 1354, 1310, 1266, 1241, 1166, 1091, 1040, 1017 cm
26

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1
+
+
;
ESI-MS: m/z 447.10 [M+H] ; HR-ESIMS: m/z 447.14818 calcd for C24
H
22
N
4
O
3
S+H
(447.15126).
4
.8.33. N-(4-Acetylphenyl)-N-((1-(2-cyanophenyl)-1H-1,2,3-triazol-4-yl)methyl)benzene
1
sulfonamide (26w). White solid; m.p. 136-138 °C; H NMR (CDCl 400 MHz): δ 8.21 (s, 1H,
3
,
Ar-CH), 7.86 (dd, J = 11.5, 8.5 Hz, 3H, Ar-CH), 7.79 (d, J = 3.9 Hz, 2H, Ar-CH), 7.68-7.58 (m,
4
H, Ar-CH), 7.50 (t, J = 7.7 Hz, 2H, Ar-CH), 7.29 (d, J = 8.5 Hz, 2H, Ar-CH), 5.06 (s, 2H, N-
13
2 3
CH ), 2.57 (s, 3H, COCH ); C NMR (CDCl3, 125 MHz): δ 197.22, 144.25, 143.15, 138.26,
1
1
1
4
37.66, 136.14, 134.37, 133.37, 129.79, 129.37, 129.21, 128.15, 127.62, 125.44, 123.85, 115.47,
06.78, 46.29, 26.70; IR (CHCl ): νmax 3860, 3848, 3791, 3683, 3438, 3141, 2230, 2139, 1682,
3
-1
600, 1505, 1446, 1408, 1356, 1310, 1293, 1266, 1243 1164, 1091, 1015 cm ; ESI-MS: m/z
+
+
58.10 [M+H] ; HR-ESIMS: m/z 458.12826 calcd for C H N O S+H (458.13086).
2
4
19
5
3
4
.8.34. N-(4-Acetylphenyl)-N-((1-(2-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)
1
benzene sulfonamide (26x). White solid; m.p. 142-144 °C; H NMR (CDCl 400 MHz): 7.84
3
,
(m, 4H, Ar-CH), 7.67 (m, 5H, Ar-CH), 7.49 (m, 3H, Ar-CH), 7.24-7.23 (m, 2H, Ar-CH), 5.06
1
3
(
s, 2H, N-CH ), 2.56 (s, 3H, COCH ); C NMR (CDCl , 100 MHz) δ 197.06, 143.18, 143.08,
2 3 3
1
37.78, 136.06, 134.54, 133.29, 133.08, 130.60, 129.18, 129.14, 128.89, 128.03, 127.60, 127.32
3
2
4
1
(
q, J = 4.9 Hz), 126.11 (q, JC-F = 30.5 Hz), 126.03 (d, J = 1.7 Hz), 122.45 (q, J
=
C-F
C-F
C-F
2
1
5
3
71.7 Hz), 46.22, 26.60; IR (CHCl ): νmax 3435, 3145, 3067, 2923, 2852, 1684, 1601, 1508,
-1
+
354, 1316, 1267, 1166, 1135, 1092, 1042 cm ; ESI-MS: m/z 501.00 [M+H] ; HR-ESIMS: m/z
+
01.12115 calcd for C H F N O S+H (501.12300).
2
4
19
3
4
3
4
.9. Cell culture and treatments. Human colorectal adenocarcinoma LS-180 cells were
purchased from ECACC, England. LS-180 cells were grown in MEM growth medium mixed in
the ratio of 1: 1. The media for the cell lines were supplemented with 1% MEM non-essential
27

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-1
amino acids along with 10% FCS, 100 U penicillin G and 100 ꢀg ml of streptomycin. Cells
were grown in 5% CO2 at 37 °C with 95% humidity. All the test compounds were dissolved in
DMSO for treatment of LS-180 cells, while the untreated control cultures received only the
vehicle (DMSO < 0.2%).
4
4
4
.10. P-gp induction activity in LS-180 cells: As described earlier [8].
.11. Cell viability studies in LS-180 cells. As described earlier [8].
6
.12. Western-blot analysis. LS180 cells were seeded at a density of 2×10 in 60mm dishes
and treated with 5 ꢀM of 6 and 20j for 24 hr. After incubation cells were collected, washed with
PBS and were lysed with lysis buffer (RIPA) composed of 0.5 mM NaVO 2 mM PMSF, 50mM
4
,
NaF and 1% protease inhibitor cocktail for 30 min on ice. The cells were vortexed and
o
centrifuged at 13000 rpm for 15min at 4 C. The supernatant were taken as whole cell lysate and
total protein was measured by Bradford reagent. The concentration of protein in each well was
7
0 ug for SDS-PAGE. Resolved proteins by SDS-PAGE were transferred to PVDF membrane
for 2 hr at 100V. Non-specific binding was blocked by blocking buffer, TBST (10 mM Tris-HCl,
50 mM NaCl and 0.1% Tween 20) containing 5% non-fat milk for 1h at room temperature. The
1
membrane was probed overnight with primary antibody followed by three times washing with
TBST for 15min. Then the membrane was incubated with horseradish peroxidase-conjugated
with rabbit secondary antibody for 1h and washed again three times with blocking buffer. The
protein bands were detected using ECL chemiluminescence on X-ray film.
4
4
.13. Neuroprotection activity. As reported earlier [9].
.14. Statistical analysis: Data are expressed as mean ± SD of three independent experiments
unless otherwise indicated. The comparisons were made between control and treated groups or
28