1,2,3-triazole tethered Indole-3-glyoxamide derivatives as multiple inhibitors of 5-LOX, COX-2 & tubulin: Their anti-proliferative & anti-inflammatory activity

Article abstract of DOI:10.1016/j.bioorg.2018.07.029

To evaluate the role of COX-2 and 5-LOX as dual inhibitors in controlling the cancer cell proliferation, a set of two series having 42 compounds of 1, 2, 3-Tethered Indole-3-glyoxamide derivatives were synthesized by employing click chemistry approach and were also evaluated for their in vitro cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX) inhibitory activities with in vivo anti-inflammatory and in vitro anti-proliferative potencies. Among the compounds tested, compounds 11q and 13s displayed excellent inhibition of COX-2 (IC₅₀ 0.12 μM) with good COX-2 selectivity index (COX-2/COX-1) of 0.058 and 0.046 respectively. Compounds 11q and 13s also demonstrated comparable 5-LOX inhibitory activity with IC₅₀ 7.73 and 7.43 μM respectively to that of standard Norhihydroguaiaretic acid (NDGA: IC₅₀ 7.31 μM). Among all the selected cell lines, prostate cancer cell line DU145 was found to be susceptible to this class of compounds. Among all the tested compounds, compounds 11g, 11i, 11k, 11q, 13r, 13s and 13u demonstrated excellent to moderate anti-proliferative activity with IC_50s ranging between 6.29 and 18.53 μM. Compounds 11q and 11g demonstrated better anti-proliferative activities against DU145 cancer cell line with IC₅₀ values 8.17 and 8.69 μM respectively when compared to the standard drug etoposide (VP16; IC₅₀ 9.80 μM). Compounds 11g, 11k, 11q, 13s and 13u showed good dual COX-2/5-LOX inhibitory potentials with excellent anti-proliferative activity. Results from carrageenan-induced hind paw edema demonstrated that compounds 11b, 11l, 11q and 13q exhibited significant anti-inflammatory activity with 69–77% inhibition at 3 h, 75–82% inhibition at 5 h when compared to the standard drug indomethacin (66.6% at 3 h and 77.94% at 5 h). Ulcerogenic study revealed that compounds 11q and 13q did not cause any gastric ulceration. In vitro tubulin assay resuted that compound 11q interfered with microtubulin dynamic and act as tubulin polymerization inhibitor. In silico molecular docking studies demonstrated that compounds 11q and 13s are occupying the colchicines binding site of tubulin polymer and 11q illustrated very good binding affinities towards COX-2 and 5-LOX.

Full text of DOI:10.1016/j.bioorg.2018.07.029

Accepted Manuscript
1
5
,2,3-triazole tethered Indole-3-glyoxamide derivatives as multiple inhibitors of
-LOX, COX-2 & Tubulin: Their anti-proliferative & anti-inflammatory activ-
ity
Fatima Naaz, M.C. Preeti Pallavi, Syed Shafi, Naveen Mulakayala, M. Shahar
Yar, H.M. Sampath Kumar
PII:
S0045-2068(18)30331-6
DOI:
https://doi.org/10.1016/j.bioorg.2018.07.029
YBIOO 2448
Reference:
To appear in:
Bioorganic Chemistry
Received Date:
Revised Date:
Accepted Date:
11 April 2018
22 July 2018
26 July 2018
Please cite this article as: F. Naaz, M.C. Preeti Pallavi, S. Shafi, N. Mulakayala, M. Shahar Yar, H.M. Sampath
Kumar, 1,2,3-triazole tethered Indole-3-glyoxamide derivatives as multiple inhibitors of 5-LOX, COX-2 & Tubulin:
Their anti-proliferative & anti-inflammatory activity, Bioorganic Chemistry (2018), doi: https://doi.org/10.1016/
j.bioorg.2018.07.029
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1
,2,3-triazole tethered Indole-3-glyoxamide derivatives as multiple inhibitors of 5-
LOX, COX-2 & Tubulin: Their anti-proliferative & anti-inflammatory activity
a
b
c
d
a
Fatima Naaz, M. C. Preeti Pallavi, Syed Shafi, * Naveen Mulakayala, M. Shahar Yar,* H. M.
b
Sampath Kumar
a
Department of Pharmaceutical Chemistry, School of Pharmaceutical Education and Reseach,
b
Jamia Hamdard, New Delhi, India; Immunochemistry lab, Natural Product division, CSIR-
c
IICT-Hyderabad, India; Department of chemistry, School of chemical and lifescience, Jamia
Hamdard, New Delhi, India, ISVAK life Sciences, Pragati Nagar, Hyderabad 500090, India.
d
Corresponding authors. Tel.: +91 9990806873, +91 9811811033; fax: +91 91 01126059663.
E-mail addresses: syedshafi@jamiahamdard.ac.in (SS), msyar@jamiahamdard.ac.in (MSY)
Abstract:
To evaluate the role of COX-2 and 5-LOX as dual inhibitors in controlling the cancer cell
proliferation, a set of two series having 42 compounds of 1, 2, 3-Tethered Indole-3-glyoxamide
derivatives were synthesized by employing click chemistry approach and were also evaluated
for their in vitro cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-
LOX) inhibitory activities with in vivo anti-inflammatory and in vitro anti-proliferative
potencies. Among the compounds tested, compounds 11q and 13s displayed excellent inhibition
of COX-2 (IC50 0.12 µM) with good COX-2 selectivity index (COX-2/COX-1) of 0.058 and
0
.046 respectively. Compounds 11q and 13s also demonstrated comparable 5-LOX inhibitory
activity with IC50 7.73 and 7.43 µM respectively to that of standard Norhihydroguaiaretic acid
NDGA: IC50 7.31 µM). Among all the selected cell lines, prostate cancer cell line DU145 was
found to be susceptible to this class of compounds. Among all the tested compounds, compounds
1g, 11i, 11k, 11q, 13r, 13s and 13u demonstrated excellent to moderate anti-proliferative
(
1
activity with IC50s ranging between 6.29-18.53 µM. Compounds 11q and 11g demonstrated
better anti-proliferative activities against DU145 cancer cell line with IC50 values 8.17 and 8.69
µM respectively when compared to the standard drug etoposide (VP16; IC50 9.80 µM).
Compounds 11g, 11k, 11q, 13s and 13u showed good dual COX-2/5-LOX inhibitory potentials
with excellent anti-proliferative activity. Results from carrageenan-induced hind paw edema
demonstrated that compounds 11b, 11l, 11q and 13q exhibited significant anti-inflammatory
activity with 69-77% inhibition at 3h, 75-82% inhibition at 5h when compared to the standard
drug indomethacin (66.6% at 3h and 77.94% at 5h). Ulcerogenic study revealed that compounds
1
1
1q and 13q did not cause any gastric ulceration. In vitro tubulin assay resuted that compound
1q interfered with microtubulin dynamic and act as tubulin polymerization inhibitor. In silico
molecular docking studies demonstrated that compounds 11q and 13s are occupying the
colchicines binding site of tubulin polymer and 11q illustrated very good binding affinities
towards COX-2 and 5-LOX.
Keywords: Indole-3-glyoxamide derivatives, 1,2,3-triazoles, COX-1; COX-2; 5-LOX; Tubulin; Anti-
proliferation.
1

1
. Introduction
Worldwide cancer is the leading cause for death. About 90.5 million people were diagnosed with
cancer in 2015 which lead to 8.8 million deaths (15.7%). Cancer cases are increasing very vastly
throughout the world that it has become a very common word with one out of each family
diagnosed with cancer. Over the next 2 decades, the number of new cases is expected to rise by
about 70% [1]. There are various targets to treat cancer. Among all targets, the most fruitful for
cancer treatment is tubulin. Microtubules are synthesized by polymerization of α and β- tubulin
dimers. These microtubulins are mainly involved in the mitosis and meiosis which lead to cell
division [2]. There are various binding sites for inhibitors to bind tubulin i.e colchicine, taxane
paclitaxel and vinca (vinca alkaloids) binding sites [2-4] among which we focused on the
molecules that binds at colchicines binding site.
Recent studies regarding the relationship between arachidonic acid (AA) cascade and
carcinogenesis are revealing novel molecular targets for cancer treatment [5]. From literature, it
has been reported that many studies shows COX-2/5-LOX dual inhibition might be a suitable
adjuvant therapy to prevent liver metastasis in human ductal pancreatic adenocarcinoma [6].
Recently, it has been demonstrated that 5-LOX plays an important role in regulating cellular
proliferation as they are linked with each other through their common oxidative processes. As
overexpression of platelet 12-lipoxygenase (p12-LOX), 5-LOX enzyme and 15-LOX metabolite
leads to prostate cancer, pancreatic cancer, and breast cancer respectively. There are various
drugs such as MK591 and MK866 (in fig.1) which not only inhibit 5-LOX enzymes but also
cause cell death in prostate cancer cells [7]. Compound 1 (in fig.1) showed very mild inhibition
against COX-1 with IC50 89.53 µM and against COX-2 with IC50 87.71 µM but also exhibited
most pronounced anti-inflammatory activity and displayed effective antiproliferative activity
with growth inhibition of GI50 (MG-MID) 15.13 µM and LC50 (MG-MID) 7.58 µM [8]. From
literature, it is found that compound 2 (in fig.1), which is inhibiting 5-LOX (IC50 36.65 µM) also
showed a good anti-proliferative activity with LC50 6.49 µM [9] against A. salina species.
Similarly compound 3 (in fig. 1) also exhibited the cancer growth inhibition by inhibiting the
inflammatory markers COX-2 (IC50 0.18 µM with a good COX-1/COX-2 selectivity) and 5-
LOX (IC50 0.71 µM). It was found to induce apoptosis and G2/M phase cell cycle arrest in
human lung cancer A549 [10].
Figure 1
2

Compound 4 showed good anti-inflammatory activity and anti-proliferative activity against
DU145 cancer cell line with IC50 14.15 µM [11]. An indole derivative compound 5 was also
found to exhibit the dual action of anti-proliferative and anti-inflammatory activity and strongly
bind to the active site of the COX-2 binding site [12]. Furthermore, indoyloxy derivative 6
(
Shown in fig.1) displayed active anti-oxidant activity, anti-inflammatory activity and anti-
cancer activity against MCF-7 cancer cell line with GI50 30.84µM [13].
Indole is a buzzword in the field of research due to their wide spectrum of biological activities
which include anti-inflammatory, anti-proliferative, anti-viral, anti-HIV, anti-hypertensive, anti-
leukemic, anti-asthmatic, and also used in various sexual disorders [14]. There are various
reported natural and synthetic molecules which bind to tubulin, thereby causing an
inhibition of its polymerization and alteration in the tubulin-microtubule equilibrium such as
colchicines [15],[16], Combretastatin A-4 [15],[16], Indolyl chalcone [16],[17],
Phenylcinnamides [16], Bis (indolyl) hydrazide-hydrazone [17] indibulin (anti-tubulin agent in
clinical trial) [18].
Keeping in view the involvement of COX-2 and 5-LOX mechanisms in controlling cancer cell
proliferation and the importance of indole moiety as an attractive ligand for the development of
chemotherapeutic agents, 1,2,3-triazole tethered indole-3-glyoxamide derivatives have been
designed as dual inhibitors of COX-2 and 5-LOX for cancer chemotherapy by employing a
“hybrid conjugation of bio-active ligands” approach (fig. 2). Herein we are reporting design,
synthesis, dual inhibition of COX-2 & 5-LOX, in vivo anti-inflammatory activity and in vitro
anti-proliferative activities of 1,2,3-triazole tethered indole-3-glyoxamide derivatives.
Figure 2
2
. Materials and methods
2
.1. General
All the chemicals and reagents used in the study were purchased from Merck (India),
Spectrochem, and Sigma Aldrich which were of reagent grade. Melting points of all the
synthesized compounds were measured using Buchi labortechnik AG 9230 automated melting
point apparatus (Switzerland); IR spectra were recorded as potassium bromide pellets on a
1
Perkin Elmer 1650 spectrophotometer (USA), H- NMR spectra were determined on a Bruker
3

(
300 and 400 MHz) spectrometer and chemical shifts were expressed as ppm against TMS as
internal reference. Mass spectras were recorded on ESI-MS and Column Chromatography was
performed on (Merck) Silica gel 60 (particle size 0.06-0.20 mm). All compounds prepared in this
paper are novel and confirmed with spectral data.
2
2
.2. Chemistry
.2.1. Procedure for the synthesis of 2-(1H-indol-3-yl)-2-oxoacetyl chloride (8)
To the diethyl ether (60 mL) solution of Indole (6 g, 1.equiv), oxalylchloride (6 mL, 1.equiv)
was added drop wise over the period of 15 min and the reaction mixture was stirred at room
temperature for further 10 min to afford the yellow precipitate. The compound formed was
filtered off and washed with diethyl ether and dried under high vacuum pump to obtain the title
compound 8 in quantitative yields. Compound 8 was immediately taken for further reaction.
2
.2.2. Procedure for the synthesis of 2-(1H-indol-3-yl)-2-oxo-N-(prop-2-ynyl)acetamide (9)
Compound 8 (4 g, 1 equiv) was dissolved in dry pyridine (20 mL) then propargyl amine (1.6 mL,
.2 equivalent) was added in portions and reaction mixture was stirred at room temperature for 8
1
hrs. Completion of the reaction was monitored by TLC, the reaction mixture was poured into ice
to form the off white coloured solid. This off white coloured solid was filtered off and washed
with excess of water and vacuume dried to form 2-(1H-indol-3-yl)-2-oxo-N-(prop-2-
ynyl)acetamide in quantitative yields (4.34 g).
2
.2.3. General procedure for the click chemistry reaction (formation of 1,2,3-triazole ring 11a-
1u and 13a-13u):
1
Terminal alkyne (compound 9 or compound 12) was dissolved in 20 mL of tert. Butanol:water
1:1) solvent at ambient temperature. Then CuSO O (1.5 eq to terminal alkyne) was added to
(
4
.
5
H
2
the reaction mixture and stirred for 10 min. Then sodium ascorbate (5 eq.) was added at and
allowed to stir for 15 min. Reaction mixture colour was changed from blue to dark brown. After
1
5 min, different substituted azide (1.2 eq.) was added to reaction mixture and was allowed to
stir for further 8 h at ambient temperature. Reaction was monitored by TLC. After the
completion of the reaction, reaction mixture was quenched with water and extracted with ethyl
4

acetate. Combined organic layers were dried over anhydrous sodium sulphate, filtered and
concentrated under reduced pressure to obtain the final product.
2
-(1H-indol-3-yl)-N-((1-(4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxoacetamide (11a)
o
-1
Brown solid; 98% yield; mp 188.1 C; IR (KBr) (cm ): 3432, 3339, 3164, 3142, 2944, 1784,
1
1
633, 1615, 1530, 1515, 1481, 1430, 1334, 1296, 1101, 1029, 876, 801, 721; H NMR (400
MHz, DMSO-d
6
) δ (ppm): 12.2 (s, 1H,), 9.32 (s, 1H), 8.90 (s, 1H), 8.78 (s, 1H), 8.41 (d, 2H, J =
8
.40 Hz), 8.22 (d, 3H, J = 7.20 Hz), 7.51 (d, 1H, J = 3.20 Hz), 7.34 -7.24 (m, 2H), 4.58 (d, 2H,
1
3
J = 4.80 Hz); C NMR (100 MHz, DMSO-d
6
) : δ 181.5, 163.5, 146.5, 146.4, 140.8, 138.5,
1
1
36.2, 126.1, 125.5, 123.4, 122.5, 121.5, 121.2, 120.4, 112.5, 112.1, 34.25; MS (ESI) m/z [M +
+
] 391.0; Anal. Calcd for C19
H
14
N
6
4
O : C, 58.46; H, 3.61; N, 21.53%, Found: C, 58.49; H, 3.59;
N, 21.55%.
-(1H-indol-3-yl)-N-((1-(3-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxoacetamide (11b)
2
o
-1
Brown solid; 97% yield, mp 176.3 C; IR (KBr) (cm ) : 3442, 3323, 3132, 2974, 2873, 2813,
1
2
730, 1774, 1676, 1617, 1530, 1491, 1427, 1345, 1233, 1129, 1047, 888, 807, 733; H NMR
(
300 MHz, DMSO-d ) δ (ppm): 12.29 (s, 1H), 9.34 (t, 1H, J = 5.70), 8.76 (d, 1H, J = 1.20 Hz),
6
8
7
.65-8.63 (m, 1H), 8.52 (s, 1H), 8.24-8.20 (m, 3H), 7.73-7.68 (m, 1H), 7.55-7.52 (m, 1H), 7.28 -
+
.25 (m, 2H), 4.60 (d, 2H, J = 5.70 Hz,); MS (ESI) m/z [M + 1] 391.4; Anal. Calcd for
C
19
H
14
N
6
O
4
: C, 58.46; H, 3.61; N, 21.53%, Found: C, 58.42; H, 3.65; N, 21.52%.
N-((1-(2-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11c)
o
-1
Light Brown solid; 83% yield; mp 66.5 C; IR (KBr) (cm ): 3474, 3461, 3034, 2935, 2867,
1
2
833, 1703, 1672, 1622, 1493, 1423, 1233, 1126, 1046, 892, 747, 643; H NMR (400 MHz,
CDCl
3
) δ ppm: 9.54 (s, 1H), 9.11 (d, 1H, J = 3.20 Hz), 8.39-8.377 (m, 1H), 8.12 (t, 1H, J = 5.60
Hz), 8.01 (s, 1H), 7.59 -7.55 (m, 2H), 7.47-7.41 (m, 3H), 7.33 -7.28 (m, 2H), 4.80 (d, 2H, J =
1
3
6
.4 Hz); C NMR (100 MHz, CDCl
3
) : δ 179.5, 162.1, 143.5, 138.0, 135.3, 134.2, 130.4, 130.3,
1
28.1, 127.4, 127.3, 126.1, 123.9, 123.7, 122.9, 121.9, 112.7, 111.2, 34.2; MS (ESI) m/z [M + 1]
+
3
80.2; Anal. Calcd for C19
H14ClN
5
2
O : C, 60.08; H, 3.72; Cl, 9.33; N, 18.44%, Found: C, 60.11;
H, 3.74; Cl, 9.31; N, 18.41%.
N-((1-(3-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11d)
5

-1
Yellow solid; 76% yield; mp 156.4 °C; IR (KBr) (cm ): 3441, 3386, 3369, 3137, 3020, 2969,
1
1
1
1
741, 1683, 1488, 1431, 1341, 1237, 1098, 1055, 782, 749; H NMR (300 MHz, DMSO-d
6
) : δ
2.26 (s, 1H), 9.30 (t, 1H, J = 5.70 Hz), 8.80 (s, 1H), 8.79 (s, 1H), 8.25-8.22 (m, 1H), 8.06 (s,
H), 7.94 (d, 1H, J = 7.80 Hz), 7.64-7.54 (m, 3H), 7.28 -7.25 (m, 2H), 4.57 (d, 2H, J = 5.7 Hz);
+
MS (ESI) m/z [M + 1] 380.2; Anal. Calcd for C19
H14ClN
5
2
O : C, 60.08; H, 3.72; Cl, 9.33; N,
1
8.44%, Found: C, 60.09; H, 3.68; Cl, 9.37; N, 18.42%.
N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11e)
o
-1
Light yellow solid; 70% yield; mp 215.8 C; IR (KBr) (cm ): 3428, 3391, 3189, 3143, 3021,
1
2
1
1
4
967, 1612, 1497, 1430, 1238, 1054, 891, 748, 644; H NMR (400 MHz, DMSO-d
6
) δ (ppm):
2.22 (s, 1H), 9.25 (t, 1H, J = 6.00 Hz), 8.77 (d, 1H, J = 2.80 Hz), 8.70 (s, 1H), 8.23-8.21 (m,
H), 7.94 (d, 2H, J = 8.80 Hz), 7.64 (d, 2H, J = 8.80 Hz), 7.53 -7.51 (m, 1H), 7.27-7.22 (m, 2H),
1
3
.55 (d, 2H, J = 6.00 Hz); C NMR (100 MHz, DMSO-d
6
): δ 181.6, 163.5, 145.8, 138.5, 136.2,
1
35.4, 132.7, 129.8, 126.1, 123.4, 122.5, 121.6, 121.2, 112.5, 112.1,34.2; MS (ESI) m/z [M + 1]
+
3
80.2; Anal. Calcd for C19
H14ClN
5
2
O : C, 60.08; H, 3.72; Cl, 9.33; N, 18.44%, Found: C, 60.04;
H, 3.74; Cl, 9.29; N, 18.43%.
N-((1-(2-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11f)
o
-1
Yellow solid; 82% yield; mp 178.2 C; IR (KBr) (cm ): 3400, 3308, 3271, 3241, 3209, 3142,
1
2
1
999, 1632, 1614, 1497, 1237, 1149, 1058, 801, 754; H NMR (400 MHz, DMSO-d
6
) δ (ppm):
2.23 (s, 1H), 9.28 (t, 1H, J = 5.60 Hz), 8.75 (d, 1H, J = 2.8 Hz), 8.45 (s, 1H), 8.22-8.20 (m, 1H,
m), 7.81 (t, 1H, J = 7.60 Hz), 7.59-7.50 (m, 3H), 7.41 (t, 1H, J = 7.60 Hz), 7.27 -7.21 (m, 2H),
1
3
4
1
1
6
.57 (d, 2H, J = 6.00 Hz); C NMR (100 MHz, DMSO-d ) : δ 181.7, 163.6, 145, 138.5, 136.2,
6
31.5, 125.8 , 124.2, 123.4, 122.5, 121.2, 117.2, 117.0, 112.5, 112.1, 34.11; MS (ESI) m/z [M +
+
]
364.2; Anal. Calcd for C19
H14FN
5
2
O : C, 62.81; H, 3.88; F, 5.23; N, 19.27%, Found: C,
2.84; H, 3.91; F, 5.18; N, 19. 26%.
N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11g)
o
-1
Cream Brown solid; 88% yield; mp 148.6 C; IR(KBr) (cm ): 3475, 3392, 3239, 3184, 3074,
1
2
986, 2938, 1653, 1613, 1512, 1243, 1098, 833, 661cm-1; H NMR (300 MHz, DMSO-d
6
) δ
(
ppm): 12.26 (s, 1H), 9.29 (brs, 1H), 8.80 (s, 1H), 8.68 (d, 1H, J = 3.00 Hz), 8.24-8.23 (m, 1H),
6

7
.97-7.95 (m, 2H), 7.55-7.41 (m, 3H), 7.27 (d, 2H, J = 3.60 Hz), 4.57 (d, 2H, J = 3.00 Hz); MS
+
(
ESI) m/z [M + 1] 364.1; Anal. Calcd for C19
H14FN
5
2
O : C, 62.81; H, 3.88; F, 5.23; N, 19.27%,
Found: C, 62.77; H, 3.89; F, 5.21; N, 19. 31%.
N-((1-(2-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11h)
o
-1
Brown solid; 89% yield; mp 64.1 C; IR (KBr) (cm ): 3433, 3354, 3055, 3025, 2982, 1704,
1
1
681, 1623, 1559, 1488, 1458, 1287, 1237, 1177, 1156, 1133, 1099, 1019, 889, 751; H NMR
(
400 MHz, CDCl
3
) δ (ppm): 10.14 (s, 1H), 9.11 (d, 1H, J = 3.20 Hz), 8.39 – 8.36 (m, 1H), 8.10
(
s, 1H), 7.74-7.72 (dd, 1H, J = 8.00 and 1.20 Hz), 7.47-7.39 (m, 2H), 7.30-7.24 (m, 2H), 7.10 -
1
3
7
1
1
.05 (m, 2H), 4.78 (d, 2H, J = 6.00 Hz), 3.85 (s, 3H); C NMR (100 MHz, CDCl ) : δ 179.6,
3
62.1, 150.6, 138.3, 135.5, 129.8, 126.2, 125.5,125.0, 123.9, 123.5, 122.7, 121.8, 120.7, 112.6,
+
11.7, 111.4, 55.4, 34.2; MS (ESI) m/z [M + 1] 376.2; Anal. Calcd for C20
H
17
N
5
3
O : C, 63.99;
H, 4.56; N, 18.66%, Found: C, 64.03; H, 4.51; N, 18.68%.
N-((1-(3,4,5-trimethoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide
11i)
(
o
-1
Yellow solid; 83% yield; mp 281 C; IR (KBr) (cm ): 3412, 3141, 30644, 2948, 1673, 1632,
1
1
606, 1472, 1268, 1235, 1185, 1155, 1129, 1073, 1058, 863, 753, 652; H NMR (400 MHz,
DMSO-d
6
) δ (ppm): 12.23 (s, 1H), 9.26 (t, 1H, J = 5.60 Hz), 8.77 (d, 1H, J = 3.2 Hz), 8.69 (s,
1
H), 8.22 (d, 1H, J = 5.60 Hz), 7.53-7.51 (m, 1H), 7.25-7.22 (m, 2H), 7.18 (s, 2H), 4.55 (d, 2H, J
1
3
=
6.00 Hz), 3.85 (s, 6H), 3.68 (s, 3H); C NMR (100 MHz, DMSO-d ) : δ 181.6, 163.5, 153.4,
6
1
5
1
45.4, 138.5, 137.2, 136.2, 132.5, 126.1, 123.4, 122.5, 121.4, 121.2, 112.5, 112.1, 97.9, 60.1,
+
6.2, 34.2; MS (ESI) m/z [M + 1] 436.3. Anal. Calcd for C22
H
21
N
5
5
O : C, 60.68; H, 4.86; N,
6.08%; Found: C, 60.73; H, 4.83; N, 16.12%.
N-((1-(4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11j)
o
-1
White solid; 94% yield; mp 211.3 C; IR (KBr) (cm ): 3412, 3409, 3155, 2946, 1772, 1660,
1
1
628, 1618, 1264, 1197, 1160, 1139, 1047, 1012, 1010, 848, 778, 654; H NMR (300 MHz,
DMSO-d
6
) δ (ppm): 12.25 (s, 1H), 9.27 (s, 1H), 8.79 (s, 1H), 8.63 (s, 1H), 8.25-8.22 (m, 1H),
7
.78 (d, 2H, J = 8.40 Hz), 7.54-7.50 (m, 1H), 7.38 (d, 2H, J = 8.10 Hz), 7.26 (t, 2H, J = 3.90
7

+
Hz), 4.57 (d, 2H, J = 5.40 Hz), 2.37 (s, 3H); MS (ESI) m/z [M + 1] 360.4. Anal. Calcd for
: C, 55.84; H, 3.77; N, 15.09 %; Found: C, 55.87; H, 3.72; N, 15.11%.
C
20
H
17
N
5
O
2
N-((1-(3,4-dimethylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide
11k)
(
o
-1
Yellow solid; 89% yield; mp 162 C; IR (KBr) (cm ): 3485, 3456, 3366, 3278, 3142, 2904,
1
1
715, 1667, 1620, 1489, 1438, 1239, 1160, 1131, 1058, 874, 746; H NMR (400 MHz, CDCl
3
) δ
(
ppm): 9.21 (s, 1H), 9.10 (d, 1H, J = 3.20 Hz), 8.42 -8.39 (m, 1H), 8.07 (t, 1H, J = 6.00 Hz), 7.95
s, 1H), 7.49-7.45 (m, 2H), 7.41-7.39 (m, 1H), 7.35-7.29 (m, 2H), 7.23 (s, 1H), 4.77 (d, 2H, J =
(
1
3
6
1
1
1
.00 Hz), 2.33 (s, 3H), 2.31 (s, 3H); C NMR (100 MHz, CDCl ) : δ 179.6, 162.0, 137.9, 137.7,
3
37.1, 135.2, 134.3, 130.1, 126.1, 123.7, 122.9, 121.9, 121.2, 120.0, 117.4, 112.8, 111.1, 34.3,
+
9.4, 18.9; MS (ESI) m/z [M + 1] 374.4, Anal. Calcd for C21
H
19
N
5
2
O : C, 64.55; H, 4.13; N,
6.76%, Found: C, 64.49; H, 4.16; N, 16.78%.
N-((1-(4-tert-butylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11l)
o
-1
Brown solid; 96% yield; mp 120.7 C; IR (KBr) (cm ): 3473, 3455, 3098, 3047, 2955, 1723,
1
1
9
618, 1554, 1498, 1231, 1078, 867, 752, 648; H NMR (400 MHz, DMSO-d
6
) : δ 12.25 (s, 1H),
.26 (t, 1H, J = 5.20 Hz), 8.78 (s, 1H), 8.62 (s, 1H), 8.22 (d, 1H, J = 5.60 Hz, ArH), 7.79 (d, 2H,
J = 8.00 Hz,), 7.56 (d, 2H, J = 8.40 Hz), 7.52 (d, 1H, J = 6.00 Hz) 7.25 (t, 2H, J = 3.60 Hz), 4.55
1
3
(
d, 2H, J = 5.20 Hz), 1.29 (s, 9H); C NMR (100 MHz, DMSO-d
6
) : δ 181.7, 163.5, 151.1,
1
1
5
45.4, 138.5, 138.3, 136.2, 134.3, 126.5, 126.1, 123.4, 122.5, 121.5, 121.2, 121, 119.6, 112.5,
+
12.1, 34.4, 34.2, 30.9; MS (ESI) m/z [M + 1] 402.1. Anal. Calcd for C23
H
23
N
5
2
O : C, 68.81; H,
.77; N, 17.44%; Found: C, 68.78; H, 5.81;; N, 17.46%.
N-((1-(4-cyanophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11m)
o
-1
Brown solid; 94% yield; mp 235.2 C; IR (KBr) (cm ): 3455, 3427, 3417, 3219, 3015, 2987,
1
1
716, 1683, 1457, 1435, 1238, 1045, 751, 658; H NMR (400 MHz, DMSO-d
6
) δ (ppm): 12.27
(
s, 1H), 9.30 (t, 1H, J = 6.00 Hz), 8.85 (s, 1H), 8.77 (s, 1H), 8.23-8.21 (m, 1H), 8.16-7.99 (m,
1
3
4
H), 7.53-7.51 (m, 1H), 7.27 -7.22 (m, 2H), 4.57 (d, 2H, J = 5.60 Hz); C NMR (100 MHz,
DMSO-d
6
) : δ 181.3, 163.5, 151.1, 142.7, 138.5, 136.2, 134.2, 124.1, 122.5, 121.3, 121.2, 120.3,
8

+
1
3
12.5, 112.1, 34.28; MS (ESI) m/z [M + 1] 371.3; Anal. Calcd for C20
H
14
N
6
2
O : C, 64.86; H,
.81; N, 22.69%; Found: C, 64. 89; H, 3.79; N, 22.71%.
N-((1-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-
oxoacetamide (11n)
o
-1
Yellow solid; 86% yield; mp 188.5 C; IR (KBr) (cm ): 3494, 3464, 3378, 3056, 2933, 1784,
1
1
666, 1618, 1489, 1435, 1323, 1241, 1165, 1123, 1061, 887, 754, 700; H NMR (400 MHz,
DMSO-d
6
) δ (ppm): 12.24 (s, 1H), 9.28 (t, 1H, J = 6.00 Hz), 8.87 (s, 1H), 8.78 (d, 1H, J = 2.80
Hz), 8.27-8.21 (m, 3H), 7.83-7.81 (m, 2H), 7.53-7.51 (m, 1H), 7.27 -7.23 (m, 2H), 4.57 (d, 2H, J
1
3
=
6.00 Hz); C NMR (100 MHz, DMSO-d
6
): δ 181.5, 163.5, 146, 138.5, 137, 136.2, 131.2,
+
1
4
26.1, 125, 123.8, 123.4, 122.5, 121.4, 121.2, 116.5, 112.5, 112.1, 34.2; MS (ESI) m/z [M + 1]
14.1; Anal. Calcd for C20
H
14
F
3
N
5
O
2
: C, 58.11; H, 3.41; F, 13.79; N, 16.94%; Found: C, 58.09;
H, 3.43; F, 13.74; N, 16.97%.
-(1H-indol-3-yl)-2-oxo-N-((1-(pyridin-3-yl)-1H-1,2,3-triazol-4-yl)methyl)acetamide (11o)
2
o
-1
Reddish brown solid; 81% yield; mp 214.6 C; IR (KBr) (cm ): 3494, 3453, 3300, 2924,
1
1
746.94, 1719, 1667, 1618, 1555, 1522, 1489, 1432, 1235, 1123, 1050, 745, 690; H NMR (400
MHz, DMSO-d
6
) δ (ppm): 12.24 (s, 1H), 9.30 (t, 1H, J = 6.00 Hz), 9.13 (s, 1H), 8.78 (d, 2H, J =
2
.8 Hz), 8.66 (d, 1H, d, J = 3.60 Hz), 8.32 (d, 1H, J = 8.00 Hz), 8.23-8.21 (m, 1H), 7.64 -7.60
1
3
(
m, 1H), 7.53-7.50 (m, 1H), 7.27-7.22 (m, 2H), 4.57 (d, 2H, J = 6.00 Hz); C NMR (100 MHz,
DMSO-d
6
) : δ 181.3, 163.5, 149.5, 145.9, 141.1, 138.5, 136.2, 127.8, 126.1, 124.5, 123.4, 122.5,
+
1
6
21.5, 121.2, 112.5, 112.1, 34.2; MS (ESI) m/z [M + 1] 347.4; Anal. Calcd for C18
H
14
N
6
2
O : C,
2.42; H, 4.07; N, 24.27%; Found: C, 62.39; H, 4.06; N, 24.29%.
N-((1-(2-chloropyridin-3-yl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide
11p)
(
o
-1
Brown solid; 88% yield; mp 148.6 C; IR (KBr) (cm ): 3449, 3381, 3245, 3115, 3098, 3016,
1
1
746, 1678, 1618, 1574, 1522, 1489, 1427, 1131, 1040, 886, 747, 692; H NMR (300 MHz,
DMSO-d
6
) : δ 12.29 (s, 1H), 9.34 (t, 1H, J = 5.40 Hz), 8.76 (d, 1H, J = 2.70 Hz), 8.65-8.63 (m,
1
H), 8.52 (s, 1H), 8.24-8.20 (m, 2H), 7.72- 7.68 (m, 1H), 7.55-7.53 (m, 1H), 7.28-7.25 (m, 2H),
9

+
4
5
.60 (d, 2H, J = 5.70 Hz); MS (ESI) m/z [M + 1] 381.8; Anal. Calcd for C18
H13ClN
6
2
O : C,
6.78; H, 3.44; Cl, 9.31; N, 22.07%; Found: C, 56.81; H, 3.42; Cl, 9.33; N, 22.04%.
N-((1-(4-ethylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11q)
o
-1
Yellow solid; 97% yield; mp 221.1 C; IR (KBr) (cm ): 3423, 3341, 3298, 3329, 1715, 1699,
1
1
7
2
680, 1523, 1489, 1439, 1240, 1132, 1057, 828, 744; 1H NMR (400 MHz, DMSO-d ) δ (ppm):
6
2.23 (s, 1H), 9.25 (t, 1H, J = 5.60 Hz), 8.78 (d, 1H, J = 2.8 Hz), 8.62 (s, 1H), 8.23-8.21 (m, 1H),
.78 (d, 2H, J = 8.40 Hz), 7.52 (d, 1H, J = 6.00 Hz), 7.39 (d, 2H, J = 8.00 Hz), 7.25 -7.24 (m,
1
3
H), 4.55 (d, 2H, J = 6.00 Hz), 2.65 (q, 2H, J = 7.60 Hz), 1.19 (t, 3H, J = 7.60 Hz) ; C NMR
(
100 MHz, DMSO-d
6
): δ 181.7, 163.5, 145.4, 144.3, 138.5, 136.2, 134.5, 129, 126.1, 123.4,
+
1
22.5, 121.2, 121, 119.9, 34.2, 27.6, 15.4; MS (ESI) m/z [M + 1] 374.4; Anal. Calcd for
C
21
H
19
N
5
2
O : C, 67.55; H, 5.13; N, 18.76%; Found: C, 67.57; H, 5.11; N, 18.77%.
N-((1-(3,4-dichlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide
11r)
(
o
-1
White solid; 89% yield; mp 222.3 C. IR (KBr) (cm ): 3434, 3408, 3352, 3115, 3009, 1731,
1
1
680, 1667, 1439, 1354, 1241, 1135, 1059, 872, 816, 754; H NMR (400 MHz, DMSO-d
6
) δ
(
ppm): 12.24 (s, 1H), 9.30 (t, 1H, J = 6.00 Hz), 8.79-8.78 (m, 2H), 8.27 (d, 1H, J = 2.40 Hz),
.23-8.21 (m, 1H), 7.98-7.95 (dd, 1H, J = 8.80 and 2.40 Hz), 7.84 (d, 1H, J = 8.80 Hz), 7.53 -
8
7
1
1
5
1
3
.51 (m, 1H), 7.27-7.22 (m, 2H), 4.55 (d, 2H, J = 6.00 Hz); C NMR (100 MHz, DMSO-d ) : δ
6
81.5, 163.5, 146, 138.5, 136.2, 136.1, 132.3, 131.7, 130.8, 126.1, 123.4, 122.5, 121.5, 121.3,
+
21.2, 119.9, 112.5, 112.1, 34.2; MS (ESI) m/z [M ] 414; Anal. Calcd for C19
H
13Cl
2
N
5
2
O : C,
5.09; H, 3.16; Cl, 17.12; N, 16.91%; Found: C, 55.07; H, 3.19; Cl, 17.08; N, 16.88%.
3
S,4R,6S)-2-(4-((2-(1H-indol-3-yl)-2-oxoacetamido)methyl)-1H-1,2,3-triazol-1-yl)-6-
(acetoxymethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (11s)
o
-1
White solid; 72% yield; mp 111.6 C. IR (KBr) (cm ): 3496, 3465, 3302, 3263, 3228, 1753,
1
1
666, 1629, 1494, 1431, 1374, 1224, 1099, 1040, 907, 751; H NMR (400 MHz, DMSO-d
6
) δ
(
ppm): 12.24 (s, 1H), 9.25 (t, 1H, J = 6.00 Hz), 8.74 (d, 1H, J = 2.80 Hz), 8.27 (s, 1H), 8.22-
8
9
.20 (m, 1H), 7.52-7.50 (m, 1H), 7.27-7.24 (m, 2H), 6.31 (d, 1H, J = 9.20 Hz), 5.66 (t, 1H, J =
.60 Hz), 5.51 (t, 1H, J = 9.60 Hz), 5.15 (t, 1H, J = 9.60 Hz), 4.45 (d, 2H, J = 6.00 Hz) 4.35-
1
0

4
.31 (m, 1H), 4.12-4.02 (m, 2H), 2.00 (s, 3H), 1.97 (s, 3H), 1.94 (s, 3H); 1.78 (s, 3H); 13C NMR
(
100 MHz, DMSO-d ) : δ 181.7, 170, 169.5, 169.3, 168.4, 163.5, 145.2, 138.5, 136.2, 126.1,
6
1
1
23.4, 122.5, 121.9, 121.2, 112.53, 112.13, 83.7, 73.2, 72.2, 70, 67.5, 61.8, 34.1, 20.4, 20.3, 20.2,
+
9.9; MS (ESI) m/z [M + 1] 600.4; Anal. Calcd for C27
H
29
5
N O11: C, 54.09; H, 4.88; N, 11.68%,
Found: C, 54.05; H, 4.92; N, 11.71%.
3
S,4R,5R,6S)-2-(4-((2-(1H-indol-3-yl)-2-oxoacetamido)methyl)-1H-1,2,3-triazol-1-yl)-6-
(acetoxymethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (11t)
o
-1
White solid; 58% yield; mp 108.7 C; IR (KBr) (cm ): 3460, 3401, 3301, 3119, 3033, 2987,
1
2
944, 2861, 2839, 1767, 1658, 1621, 1411, 1122, 1045, 1025, 781, 749, 681, 658; H NMR (300
MHz, DMSO-d
6
): δ 12.26 (1H, s, NH), 9.24 (1H, t, J = 5.70 Hz), 8.77 (d, 1H, J = 2.70 Hz),
8
5
4
.56 (s, 1H), 8.24-8.21 (m, 1H), 7.54-7.52 (m, 1H), 7.26-7.22 (m, 2H), 6.27 (1H, d, J = 9.30 Hz),
.58 (1H, t, J = 9.0 Hz), 5.49-5.44 (m, 2H), 4.51-4.49 (m, 1H), 4.47 (d, 2H, J = 5.70 Hz), 4.24-
+
.13 (m, 2H), 2.16 (s, 3H), 2.07 (s, 3H), 1.99 (s, 3H), 1.86 (s, 3H); MS (ESI) m/z [M + 1] 600;
Anal. Calcd for C27
1.67%.
H
29
5
N O11: C, 54.09; H, 4.88; N, 11.68%, Found: C, 54.11; H, 4.84; N,
1
N-((1-(4-bromophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1H-indol-3-yl)-2-oxoacetamide (11u)
o
-1
Yellow solid; 98% yield; mp 224.8 C; IR (KBr) (cm ): 3469, 3440, 3373, 3309, 3258, 3153,
1
3
:
009, 1731, 1714, 1680, 1557, 1489, 1439, 1240, 820, 749, 643; H NMR (400 MHz, DMSO-d
6
)
δ 12.23 (s, 1H), 9.26 (t, 1H, J = 6.00 Hz), 8.78 (s, 1H), 8.70 (s, 1H), 8.22 (d, 1H, J = 5.60 Hz),
.87 (d, 2H, J = 8.80 Hz), 7.76 (d, 2H, J = 8.80 Hz), 7.51 (d, 1H, J = 6.00 Hz), 7.25-7.23 (m,
7
2
1
1
3
H), 4.55 (d, 2H, J = 6.00 Hz); C NMR (100 MHz, DMSO-d ) : δ 181.6, 163.5, 145.8, 138.5,
6
36.2, 135.8, 132.7, 126.1, 123.4, 122.5, 121.8, 121.2, 121.1, 121.1, 112.5, 112.1, 34.27; (ESI)
+
m/z [M + H] 424.1; Anal. Calcd for C19
H14BrN
5
2
O : C, 53.79; H, 3.33; Br, 18.83; N, 16.51%,
Found: C, 53.82; H, 3.31; Br, 18.79; N, 16.54%.
2
.2.4. Procedure for the synthesis of 2-oxo-N-(prop-2-ynyl)-2-(1-tosyl-1H-indol-3-yl)acetamide
(12)
To the dry pyridine solution (10ml) of compound 9 (2g), p-tosyl sulphonylchloride (10.1g,
equiv) was added in portions over a period of 10 minutes. The reaction mixture was stirred at
6
1
1

room temp for further 12 hrs. Completion of reaction was monitored by TLC and reaction
mixture was quenched with ice water. A white colour precipitate thus formed. It was filtered off,
washed with excess of water and dried under vaccum to afford the title compound in quantitative
yields. Compund 12 was directly used in Cu(I) catalysed 1,3-dipolarcycloaddition (click
reaction) reaction to obtain N-tosylindoleglyoxamide-1,2,3-triazole conjugates 13a-13u (general
method has been given in section 4.1.3).
N-((1-(4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
(13a)
o
-1
Brown solid; yield: 94% ; mp 265.9 C; IR (KBr) (cm ): 3428, 3348, 3302, 3109, 3011, 2908,
1
1
683, 1644, 1597, 1444, 1388, 1341, 1233, 1174, 1137, 1106, 1087.3, 1039, 832, 747, , 537; H
NMR (300 MHz, DMSO-d
6
) : δ 9.52 (t, 1H, J = 5.70 Hz, NH), 9.17 (s, 1H), 8.92 (s, 1H), 8.44
(
d, 2H, J = 8.10 Hz), 8.24-8.21 (m, 3H), 8.03-7.96 ( m, 3H), 7.50-7.43 (m, 4H), 4.63(d, 2H, J =
1
3
5
1
2
.70 Hz) 2.34 (s, 3H); C NMR (75 MHz, DMSO-d ): δ 182.8, 162.4, 147.1, 146.5, 141.3,
6
37.6, 133.8, 133.5, 131.1, 127.8, 126.6, 126.0, 125.7, 122.6, 122.2, 120.9, 116.1, 113.6, 34.8,
+
1.5; MS (ESI) m/z [M + 1] 545.2; Anal. Calcd for C26
H
20
N
6
6
O S: C, 57.35; H, 3.70; N, 15.43;
S, 5.89%, Found: C, 57.38; H, 3.73; N, 15.38; S, 5.86%.
N-((1-(3-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13b)
(
o
-1
Brown solid; yield: 89%; mp 105.6 C; IR (KBr) (cm ): 3483, 3217, 3151, 2978, 1680, 1652.,
1
1
534, 1441, 1380, 1352, 1293, 1136, 1090, 1043, 967, 807, 753, 664, 575; H NMR (300 MHz,
DMSO-d
6
) : δ 9.55 (t, 1H, J = 6.00 Hz), 9.19 (s, 1H), 9.00 (s, 1H), 8.74 (s, 1H), 8.42 (d, 1H, J =
8
4
1
1
.70 Hz), 8.33-8.25 (m, 3H), 8.04-7.97 ( m, 3H),7.89 (t, 1H, J = 8.10 Hz), 7.50-7.43 (m, 4H),
1
3
.63 (d, 2H, J = 5.70 Hz), 2.34 (s, 3H); C NMR (75 MHz, DMSO-d ): δ 182.6, 162.4, 149.0,
6
47.1, 146.4, 137.7, 133.8, 133.6, 132.0, 131.1, 127.8, 126.6, 126.4, 125.7, 123.5, 122.6, 125.7,
+
23.5, 122.6, 122.2, 116.1, 115.0, 113.6, 34.9, 21.5; MS (ESI) m/z [M + 1] 545.0. Anal. Calcd
for C26
H
20
N
6
6
O S: C, 57.35; H, 3.70; N, 15.43; S, 5.89%, Found: C, 57.33; H, 3.74; N, 15.37; S,
5
.88%.
N-((1-(2-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13c)
(
1
2

o
-1
Arylide yellow solid; yield: 80%; mp 61.5 C; IR (KBr) (cm ): 3478, 3378, 3298, 3098,
1
2
901,1664, 1654, 1559, 1507, 1446, 1384, 1178, 1135, 967, 758, 664, 571; H NMR (300 MHz,
DMSO-d
6
) : δ 9.52 (t, 1H, J = 5.70 Hz), 9.15 (s, 1H), 8.48 (s, 1H), 8.25 (d, 1H, J = 6.90 Hz),
8
4
1
1
5
6
.04-7.97 (m, 3H), 7.79-7.77 (dd, 1H, J = 7.80 and 1.5 Hz), 7.69-7.56 (m, 3H), 7.50-7.43 (m,
1
3
H), 4.63(d, 2H, J = 5.70 Hz), 2.34 (s, 3H); C NMR (75 MHz, DMSO-d ): δ 183.1, 162.5,
6
47.1, 144.7, 137.6, 135.0, 133.9, 133.6, 132.0, 131.1, 131.0, 128.9, 128.8, 127.8, 126.6, 125.7,
+
22.6, 116.2, 113.6, 34.8, 21.5; MS (ESI) m/z [M + 1] 534; Anal. Calcd for C26
H20ClN
5
4
O S: C,
8.48; H, 3.78; Cl, 6.64, N, 13.12; S, 6.00%, Found: C, 58.51; H, 3.77; Cl, 6.63; N, 13.15; S,
.03%.
N-((1-(3-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13d)
(
o
-1
Yellow solid; yield: 73%; mp 140.3 C; IR (KBr) (cm ) : 3304, 3151, 3127, 3054, 2902, 1653,
1
1
596, 1512, 1441, 1381, 1290, 1174, 1135, 1092, 967, 780, 715, 664, 576; H NMR (300 MHz,
DMSO-d
Hz), 8.05-7.93 (m, 4H), 7.65-7.28 (m, 7H), 4.61 (d, 2H, J = 6.00 Hz), 2.34 (s, 3H); C NMR
75 MHz, DMSO-d ): 182.9, 162.4, 147.1, 146.1, 138.1, 137.6, 134.6, 133.8, 133.6, 132.0,
31.1, 128.8, 127.8, 126.6, 125.7, 122.6, 121.9, 120.1, 118.9, 116.1, 113.6, 34.9, 21.5; MS (ESI)
6
) δ (ppm): 9.52 (t, 1H, J = 6.00 Hz), 9.19 (s, 1H), 8.83 (s, 1H), 8.26 (d, 1H, J = 7.20
1
3
(
6
1
+
m/z [M + 1] 518.2; Anal. Calcd for C26
H20ClN
5
4
O S: C, 58.48; H, 3.78; Cl, 6.64, N, 13.12; S,
6
.00%, Found: C, 58.46; H, 3.76; Cl, 6.67; N, 13.11; S, 5.97%.
N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13e)
(
o
-1
Yellow solid; yield: 65%; mp 165.3 C; IR (KBr) (cm ): 3394, 3151, 3076, 2988, 1717, 1683,
1
1
650, 1506, 1439, 1380, 1229, 1174, 1136, 1090, 1043, 968, 831, 751, 665.34, 573; H NMR
(
300 MHz, DMSO-d
6
) δ (ppm): 9.54 (t, 1H, J = 5.40 Hz), 9.19 (s, 1H), 8.78 (s, 1H), 8.26 (d, 1H,
J = 7.20 Hz), 8.04-7.94 (m, 4H), 7.65 (d, 2H, d, J = 8.70 Hz), 7.47-7.42 (m, 5H), 4.61 (d, 2H, J
1
3
=
5.70), 2.33 (s, 3H); C NMR (75 MHz, DMSO-d
6
): δ 182.9, 162.4, 147.1, 146.0, 137.6, 135.9,
1
33.8, 133.6, 133.3, 131.1, 130.0, 127.8, 126.6, 125.7, 122.6, 122.0, 121.8, 116.1, 113.6, 34.9,
1
3

+
2
1.5; MS (ESI) m/z [M + 1] 534.4; Anal. Calcd for C26
H20ClN
5
4
O S: C, 58.48; H, 3.78; Cl, 6.64,
N, 13.12; S, 6.00%, Found: C, 58.47; H, 3.81; Cl, 6.61; N, 13.16; S, 6.02%.
N-((1-(2-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
(13f)
o
-1
Reddish brown solid; yield 80%; mp 101.9 C; IR (KBr) (cm ): 3463, 3157, 2955, 2863, 1653,
1
1
599, 1511, 1443, 1382, 1290, 1227, 1175, 1136, 1090, 1043, 967, 755.08, 664.09, 575.08; H
NMR (300 MHz, DMSO-d
6
): δ 9.51 (t, 1H, J = 4.50 Hz), 9.16 (s, 1H), 8.53 (s, 1H), 8.25 (d, 1H,
J = 5.70 Hz), 8.04-7.97 (m, 3H), 7.84 (t, 1H, J = 7.50 Hz), 7.59-7.45 (m, 7H), 4.63 (2H, d, J =
1
3
4
1
1
6
6
.50 Hz) 2.34 (s, 3H); C NMR (75 MHz, DMSO-d ): δ 183.0, 162.5, 147.1, 145.2, 137.6,
6
33.9, 133.6, 131.7, 131.6, 131.1, 127.8, 126.6, 126.3, 125.9, 125.7, 125.1, 122.6, 117.7, 117.4
+
16.2, 113.6, 34.8, 21.5; MS (ESI) m/z [M + 1] 518.2; Anal. Calcd for C26
H20FN
5
4
O S: C,
0.34; H, 3.90; F, 3.67, N, 13.53; S, 6.20%, Found: C, 60.37; H, 3.88; F, 3.65; N, 13.54; S,
.23%.
N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13g)
(
o
-1
Yellow solid; yield: 88%; mp 151.2 C; IR (KBr) (cm ): 3456, 3356, 3091, 2959, 1683, 1652,
1
1
515, 1445, 1380, 1230, 1177, 1088, 1044, 968, 837, 665, 571; H NMR (300 MHz, DMSO-d
6
)
δ (ppm): 9.52 (t, 1H, J = 5.40 Hz), 9.28 (s, 1H), 8.73 (s, 1H), 8.26 (d, 1H, J = 6.90 Hz), 8.05-
1
3
7
.98 (m, 5H), 7.46-7.44 (m, 6H), 4.61 (d, 2H, J = 5.40 Hz), 2.34 (s, 3H); C NMR (75 MHz,
DMSO-d
6
): δ 182.9, 162.4, 147.1, 145.8, 137.6, 133.8, 133.6, 133.1, 127.8, 126.6, 125.7, 122.8,
+
1
22.7, 122.6, 122.0, 117.3, 117.0, 116.1, 113.6, 34.9, 21.5; MS (ESI) m/z [M + 1] 518.4; Anal.
Calcd for C26
H20FN
5
4
O S: C, 60.34; H, 3.90; F, 3.67, N, 13.53; S, 6.20%, Found: C, 60.31; H,
3
.92; F, 3.69; N, 13.51; S, 6.21%.
N-((1-(2-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-
yl)acetamide (13h)
o
-1
Dark Brown solid; yield: 90% yield; mp 71.4 C; IR (KBr) (cm ): 3392.01, 3155.74, 3098, 2932,
1
5
734, 1716, 1684, 1652, 1558, 1475, 380, 1175, 1135, 1105, 1044, 1020, 967, 812, 752, 664,
1
37; H NMR (300 MHz, CD
3
OD) δ (ppm): 9.21 (s, 1H), 8.31 (s, 1H), 8.28 (d, 1H, J = 7.20 Hz),
1
4

7
4
.97-7.89 (m, 3H), 7.64-7.61 (dd, 1H, J = 8.10 and 1.50 Hz), 7.51-7.46 (m, 1H), 7.41-7.32 (m,
H), 7.24 (d, 1H, J = 8.10 Hz), 7.10 (t, 1H, J = 7.80 Hz), 4.71 ( s, 2H), 3.88 (s, 3H), 2.34 (s, 3H);
1
3
C NMR (75 MHz, DMSO-d ): 183.2, 162.5, 152.0, 147.1, 144.3, 137.6, 133.9, 133.6, 131.1,
6
1
27.8, 126.6, 126.1, 125.7, 125.4, 122.6, 121.3, 116.2, 113.6, 113.5, 56.6, 34.8, 21.5; MS (ESI)
+
m/z [M + 1] 530.5; Anal. Calcd for C27
H
23
N
5
O S: C, 61.24; H, 4.38; N, 13.22; S, 6.05%,
5
Found: C, 61.20; H, 4.36; N, 13.18; S, 6.07%.
N-((1-(3,4,5-trimethoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-
yl)acetamide (13i)
o
-1
White solid; Yield: 85% ; mp 185.1 C; IR (KBr) (cm ): 3498, 3311, 3208, 2987, 1604, 1511,
1
1
475, 1378, 1232, 1177, 1130, 1089, 968, 851, 664, 571; H NMR (300 MHz, CD
3
OD) : δ 9.23
(
(
s, 1H), 8.55-8.36 (m, 2H), 8.06-7.91 (m, 3H), 7.40-7.35 (m, 4H), 7.10 (s, 2H), 4.57 (s, 2H), 3.91
1
3
s, 6H), 3.72 (s, 3H), 2.36 (s, 3H); C NMR (75 MHz, DMSO-d
6
): δ 183.1, 162.5, 153.9, 147.4,
1
5
45.7, 137.6, 133.9, 133.0, 131.1, 127.8, 126.6, 125.7, 122.6, 122.0, 116.2, 113.6, 98.4, 60.6,
+
6.7, 34.9, 21.5; MS (ESI) m/z [M + 1] 590.1. Anal. Calcd for C29
H
27
N
5
7
O S: C, 59.07; H, 4.62;
N, 11.88; S, 5.44%, Found: C, 59.11; H, 4.57; N, 11.91; S, 5.43%.
N-((1-(4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13j)
(
o
-1
Yellow solid; yield: 91%; mp 127.4 C; IR (KBr) (cm ): 3390, 3154, 2924, 2861, 1716, 1684,
1
1
652, 1512, 1442, 1381, 1291, 1135, 1105, 1089, 967, 751, 663, 573,; H NMR (300 MHz,
DMSO-d ) δ (ppm): 9.49 (t, 1H, J = 6.00 Hz), 9.17 (s, 1H), 8.67 (s, 1H), 8.26 (d, 1H, J = 7.20
6
Hz), 8.05-7.97 (m, 3H), 7.78 (d, 2H, J = 8.40 Hz), 7.50-7.38 (m, 6H), 4.60 (d, 2H, J = 6.00 Hz)
+
2
6
.38 (s, 3H), 2.34 (s, 3H); MS (ESI) m/z [M + 1] 514.3; Anal. Calcd for C27
H
23
N
5
4
O S: C,
3.14; H, 4.51; N, 13.64; S, 6.24%, Found: C, 63.11; H, 4.55; N, 13.69; S, 6.22%.
N-((1-(3,4-dimethylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-
yl)acetamide (13k)
o
-1
Light brown solid; yield: 87%; mp 101.1 C. IR (KBr) (cm ): 3478, 3311, 3201, 2909, 2859,
1
1
684, 1654, 1559, , 1507, 1388, 1380, 1363, 1177, 967, 665, 571; H NMR (300 MHz, CD
3
OD)
δ (ppm): 9.22 (s, 1H), 8.43 (s, 1H), 8.29 (d, 1H, J = 7.50 Hz), 7.98-7.90 (m, 2H), 7.70 (d, 1H, J
1
5

=
8.10 Hz), 7.60 (s, 1H), 7.51 (d, 1H, J = 8.10 Hz), 7.42-7.23 (m, 5H), 4.70 (s, 2H) 2.35 (s, 6H),
1
3
2
1
1
1
.24 (s, 3H); C NMR (300 MHz, DMSO-d ): 182.9, 162,1, 147.1, 145.8, 138.7, 137.6, 135.4,
6
33.7, 133.3, 131.1, 131.0, 127.8, 125.7, 125.1, 122.6, 121.5, 121.2, 117.6, 113.8, 34.8, 21.5,
+
9.8, 19.4; MS (ESI) m/z [M + 1] 528.3. Anal. Calcd for C28
H
25
N
5
4
O S: C, 63.74; H, 4.78; N,
3.27; S, 6.08%, Found: C, 63.77; H, 4.82; N, 13.22; S, 6.05%.
N-((1-(4-tert-butylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-
yl)acetamide (13l)
o
-1
White solid; yield: 89.7%; mp 91 C; IR (KBr) (cm ): 3463, 3154, 2960, 2867, 1767, 1698, 1653,
1
1
513, 1441, 1382, 1291, 1228, 1175, 1136, 1090, 1045, 968, 753, 717, 663, 574; H NMR (300
MHz, CD
3
OD): δ 9.23 (s, 1H), 8.45 (s, 1H), 8.28 (d, 1H, J = 7.20 Hz), 7.97-7.89 (m, 3H), 7.73
(
d, 2H, J = 8.70 Hz, ArH), 7.58 (d, 2H, J = 8.70 Hz), 7.48-7.25 ( m, 4H), 4.71 (s, 2H) 2.30 (s,
+
3
5
H), 1.35 (s, 9H); MS (ESI) m/z [M +1] 556.5; Anal. Calcd for C30
H
29
N
5
4
O S: C, 64.85; H,
.26; N, 12.60; S, 5.77%, Found: C, 64.91; H, 5.29; N, 12.57; S, 5.71%.
N-((1-(4-cyanophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13m)
(
o
-1
Yellow solid; yield: 88%; mp 208.4 C. IR (KBr) (cm ): 3466, 3255, 31057, 2987, 2836, 1608,
1
1
654, 1509, 1478, 1445, 1380, 1364, 1283, 1176, 1135, 1088, 1043, 967, 664, 570; H NMR
(
300 MHz, DMSO-d
6
): δ 9.53 (t, 1H, J =3.60 Hz), 9.19 (s, 1H), 8.83 (s, 1H), 8.26 (d, 1H, J =
6
.90 Hz), 8.11-7.91 (m, 6H), 7.65-7.26 (m, 4H), 7.19 (d, 1H, J = 7.80 Hz), 4.62 (d, 2H, J = 3.60
1
3
Hz), 2.34 (s, 3H); C NMR (100 MHz, DMSO-d
6
): 182.9, 167.2, 162.4, 147.1, 146.1, 138.9,
1
1
3
37.6, 134.5, 133.9, 133.6, 131.1, 129.8, 129.6, 127.8, 126.6, 125.7, 122.6, 121.8, 119.8, 119.2,
+
16.2, 113.6, 34.9, 21.5; MS (ESI) m/z [M ] 524; Anal. Calcd for C27
H
20
N
6
4
O S : C, 61.82; H,
.84; N, 16.02; S, 6.11%, Found: C, 61.87; H, 3.77; N, 16.05; S, 6.07%.
N-((1-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-
yl)acetamide (13n)
o
-1
Brown solid; yield: 81%; mp 193.2 C; IR (KBr) (cm ): 3392, 3153, 3061, 2927, 1792, 1734,
1
5
716, 1684, 1653, 1600, 1539, 1508, 1326, 1295, 1090, 1071, 1043, 1090, 967, 752, 664, 573,
1
37; H NMR (300 MHz, DMSO-d
6
) δ (ppm): 9.54 (t, 1H, J = 4.50 Hz), 9.18 (d, 1H, J = 3.00
1
6

Hz), 8.94 (s, 1H), 8.30-8.25 (m, 3H), 8.05-7.98 (m, 3H), 7.80-7.71 (m, 2H), 7.47-7.44 (m, 4H),
+
4
.62 (d, 2H, J = 4.50 Hz), 2.34 (s, 3H); MS (ESI) m/z [M + 1] 568.6. Anal. Calcd for
C
27
H
20
F
3
N
5
4
O S : C, 57.14; H, 3.55; F, 10.04; N, 12.34; S, 5.65%, Found: C, 57.09; H, 3.57; N,
9
.99; S, 5.71%.
2
-oxo-N-((1-(pyridin-3-yl)-1H-1,2,3-triazol-4-yl)methyl)-2-(1-tosyl-1H-indol-3-yl)acetamide
(13o)
o
-1
Yellow solid; yield: 79%; mp 95.6 C. IR (KBr) (cm ): 3486, 3155, 3056, 2993, 1652, 1597,
1
1
9
511, 1443, 1389, 1175, 1136, 1044, 967, 753, 663, 574; H NMR (300 MHz, CD
3
OD) δ (ppm):
.23 (s, 1H), 8.62 (s, 1H), 8.35-8.29 (m, 2H), 7.99-7.91 (m, 3H), 7.71-7.66 (m, 2H), 7.46-7.36
+
(
m, 4H) 7.26-7.21 (m, 1H), 4.73 (s, 2H), 2.36 (s, 3H); MS (ESI) m/z [M + 1] 501.2. Anal. Calcd
for C25
H
20
N
6
4
O S : C, 59.99; H, 4.03; N, 16.79; S, 6.41%, Found: C, 60.03; H, 3.98; N, 16.82; S,
6
.47%.
N-((1-(2-chloropyridin-3-yl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-
yl)acetamide (13p)
o
-1
White solid; yield: 82%; mp 116.6 C. IR (KBr) (cm ): 3491, 3151, 3060, 2975, 1652, 1511,
1
1
479, 1436, 1383, 1291, 1175, 1093, 753, 663, 575; H NMR (300 MHz, CD
3
OD) δ (ppm): 9.21
(
s, 1H), 8.58 - 8.56 (dd, 1H, J = 4.80 and 1.80 Hz), 8.42 (s, 1H), 8.29 (d, 1H, J = 7.80 Hz), 8.12-
.09 (dd, 1H, J = 7.80 and 1.80 Hz), 7.98-7.90 (m, 3H), 7.64-7.59 (m, 1H), 7.42-7.32 (m, 4H),
8
4
1
1
3
1
3
.74 (s, 2H) 2.35 (s, 3H); C NMR (75 MHz, DMSO-d ): 183.0, 162.5, 151.3, 147.1, 145.7,
6
45.0, 137.8, 137.6, 133.9, 133.6, 132.2, 131.1, 127.8, 126.6, 125.9, 125.7, 124.7, 122.6 116.2,
+
13.6, 34.8, 21.5; MS (ESI) m/z [M + 1] 535.1; Anal. Calcd for C25
H19ClN
6
4
O S : C, 56.13; H,
.58; Cl, 6.63; N, 15.71; S, 5.99%, Found: C, 56.16; H, 3.63; N, 15.67; S, 5.93%.
N-((1-(4-ethylphenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13q)
(
o
-1
Yellow solid; yield: 94%; mp 77.7 C; IR (KBr) (cm ): 3366, 3151, 3098, 2964, 1684, 1652,
1
1
597, 1513, 1476, 1379, 1291, 1228, 1135, 1105, 1089, 1045, 967, 834, 751, 663, 574; H NMR
(
300 MHz, DMSO-d
6
) δ (ppm): 9.49 (t, 1H, J = 5.70 Hz), 9.18 (s, 1H), 8.68 (s, 1H), 8.26 (d, 1H,
J = 6.90 Hz), 8.04-7.97 (m, 3H), 7.80 (d, 2H, J = 8.10 Hz), 7.45-7.40 (m, 6H), 4.61 (d, 2H, J =
1
7

1
3
5
.70 Hz), 2.67 (q, 2H, J = 7.50 Hz), 2.33 (s, 3H), 1.20 (t, 3H, J = 7.50 Hz); C NMR (300 MHz,
DMSO-d
6
): δ 183.0, 162.4, 147.1, 145.6, 144.8, 137.6, 135.0, 133.9, 133.6, 131.1, 129.5, 127.8,
+
1
26.6, 125.7, 122.6, 121.6, 120.4, 116.2, 113.6, 34.9, 28.1, 15.8; MS (ESI) m/z [M + 1] 528.4;
Anal. Calcd for C28
N, 13.32; S, 6.04%.
H
25
N
5
4
O S: C, 63.74; H, 4.78; N, 13.27; S, 6.08%, Found: C, 63.71; H, 4.85;
N-((1-(3,4-dichlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-
yl)acetamide (13r)
o
-1
Cream Brown solid; yield: 83%; mp 140.1 C; IR (KBr) (cm ): 3456, 3354, 3288, 3011, 2906,
1
1
684, 1595, 1508, 1489, 1380, 1177, 1089, 968, 812, 749, 664, 571; H NMR (300 MHz,
DMSO-d
6
) δ (ppm): 9.54 (t, 1H, J = 5.70 Hz), 9.20 (s, 1H), 8.81 (s, 1H), 8.25-8.20 (m, 2H),
8
.13-8.00 (m, 4H), 7.86 (d, 1H, J = 8.70 Hz, ArH), 7.50-7.40 (m, 4H, m), 4.60 (d, 2H, J = 5.70
1
3
Hz) 2.28 (s, 3H); C NMR (300 MHz, DMSO-d
6
) : 182.8, 162.4, 147.1, 146.2, 137.7, 136.6,
1
1
3
33.8, 133.6, 132.8, 132.2, 131.3, 131.1, 127.8, 126.6, 125.7, 122.6, 122.0, 122.0, 120.4, 116.1,
+
13.6, 34.9, 21.5. MS (ESI) m/z [M] 568.3; Anal. Calcd for C26
H
19Cl
2
N
5
4
O S: C, 54.94; H,
.37; Cl, 12.47; N, 12.32; S, 5.64%, Found: C, 55.00; H, 3.33; Cl, 12.41; N, 12.29; S, 5.66%.
(2S,4R,5S)-2-(acetoxymethyl)-6-(4-((2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamido)
methyl)-1H-
1
,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (13s)
o
-1
White solid; 70% yield; mp 114.3 C. IR (KBr) (cm ): 3435, 3233, 3179, 2993, 1761, 1664,
1
1
9
8
654, 1560, 1387, 1378, 1220, 1177, 958, 678, 556; H NMR (400 MHz, DMSO-d
6
) δ (ppm):
.48 (t, 1H, J = 6.00 Hz), 9.15 (s, 1H), 8.33 (s, 1H), 8.23 (d, 1H, J = 6.80 Hz), 8.02 (d, 2H, J =
.40 Hz), 7.96 ( d, 1H, J = 8.00 Hz), 7.47-7.40 (m, 4H), 6.33 (d, 1H, J = 9.60 Hz), 5.67 (t, 1H, J
=
9.60 Hz), 5.54 (t, 1H, J = 9.60 Hz), 5.16 (t, 1H, J = 10.00 Hz), 4.48 (d, 2H, J = 5.60 Hz), 4.37-
4
3
1
1
.33 (m,1H), 4.13-3.99 (m, 2H), 2.32 (s, 3H), 2.00 ( s, 3H), 1.97 (s, 3H), 1.93 (s, 3H), 1.78 (s,
1
3
H); C NMR (100 MHz, DMSO-d ) : δ 182.5, 170.0, 169.5, 169.3, 168.4, 161.9, 146.6, 144.9,
6
37.5, 137.2, 136.2, 133.3, 133.0, 130.6, 128.0, 127.4, 126.1, 125.4, 125.2, 123.4, 122.5, 122.0,
21.2, 115.6, 113.1, 112.5, 83.7, 73.2, 72.1, 70, 67.4, 61.8, 34.3, 21.0, 20.4, 20.3, 20.2, 19.9; MS
+
(
ESI) m/z [M + 1] 754; Anal. Calcd for C34
H
35
5
N O13S: C, 54.18; H, 4.68; N, 9.29; S, 4.25%,
Found: C, 54.22; H, 4.65; N, 9.34; 4.22%.
1
8

2
1
S,3R,4R,5S)-2-(acetoxymethyl)-6-(4-((2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamido)methyl)-1H-
,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (13t)
o
-1
White solid; yield: 67%; mp 111.2 C; IR (KBr) (cm ): 3434, 2793, 1752, 1684, 1654, 1559,
1
1
507, 1387, 1368, 1226., 1176, 968, 665, 572; H NMR (300 MHz, DMSO-d
6
): δ 9.44 (t, 1H, J
=
5.40 Hz), 9.15 (s, 1H), 8.24 (s, 2H), 8.04-7.97 (m, 3H), 7.47-7.44 (m, 4H), 6.26 (d, 1H, J =
9
2
.00 Hz), 5.62 (t, 1H), 5.47-5.43 (m, 2H), 4.59-4.51 (m,3H), 4.14-4.05 (m, 2H), 2.35 (s, 3H),
+
.18 (s, 3H), 1.98 (s, 3H), 1.94 (s, 3H), 1.83 (s, 3H); MS (ESI) m/z [M + 1] 754; Anal. Calcd
for C34
H
35
5
N O13S: C, 54.18; H, 4.68; N, 9.29; S, 4.25%, Found: C, 54.16; H, 4.71; N, 9.32;
4
.21%.
N-((1-(4-bromophenyl)-1H-1,2,3-triazol-4-yl)methyl)-2-oxo-2-(1-tosyl-1H-indol-3-yl)acetamide
13u)
(
o
-1
White solid; yield: 92%; mp 159.6 C; IR (KBr) (cm ): 3458, 3157, 3029, 2987, 1684, 1653,
1
1
507, 1497, 1478, 1445, 1384, 1190, 1177, 1135, 1104, 1089, 967, 663, 571; H NMR (300
MHz, DMSO-d
6
): δ 9.50 ( t, 1H, J = 5.70 Hz), 9.16 (s, 1H), 8.75 (s, 1H), 8.25 (d, 1H, J = 6.90
Hz) 8.03-7.96 (3H, m , ArH), 7.88 (d, 2H, J = 8.70 Hz), 7.79 (d, 2H, J = 8.40 Hz), 7.55-7.43 (m,
1
3
4
1
1
5
5
H), 4.59 (d, 2H, J = 5.70 Hz), 2.33 (s, 3H); C NMR (75 MHz, DMSO-d ): 182.9, 162.4, 147.1,
6
46.0, 137.6, 136.2, 133.8, 133.5, 133.2, 131.1, 127.8, 126.6, 125.7, 122.6, 122.3, 121.8, 121.6,
+
16.1, 113.6, 34.9, 21.5; MS (ESI) m/z [M + 2] 579.9; Anal. Calcd for C26
H20BrN
5
4
O S: C,
3.99; H, 3.49; Br, 13.81; N, 12.11; S, 5.54%, Found: C, 54.04; H, 3.46; Br, 13.79; N, 12.14; S,
.57%.
2
2
.3. Pharmacology
.3.1. MTT Assay
Anti-proliferative activity of all the synthesized compounds were evaluated against the panel of
three cancer cell lines such as ovaries (SKOV3), prostate (DU145), and cervical (HELA) cancer
3
cell line using MTT assay [19]. Cancer cells were seeded on 96-well plate (3-5×10 cells per
o
well) and incubated for 48 h at 37 C then treated with different concentration of the test
compound with respective to vehicle control. After 48 h of incubation, it was exposed to 20 µl of
freshly prepared MTT reagent (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)
1
9

incubated for 2 h. The supernatant from each well was removed and replaced with DMSO
100µl) to dissolve formazan crystal [20]. The optical density (absorbance) was recorded at 570
nm with reference wavelength of 620 nm.
(
2
.3.2. In vitro 5-LOX inhibition assay
All the synthesized compounds were evaluated for their 5-LOX inhibitory potential [20].
Solutions of the compounds were prepared at different concentration. Assay standardization was
done with NDGA (Nordihydroguaiaretic acid), a selective 5-LOX inhibitor. Blank (assay buffer
+
arachidonic acid), positive control (enzyme in buffer assay + arachidonic acid) and 10µL of
compound was added to 90 µL solution (in assay buffer) of 5-LOX enzyme (soyabean
lipoxygenase) in the wells of 96-well plate. This 96-well plate was placed on the shaker. After 5
min, 100 µL of the chromogen (developing reagent) was added to each well and was again
shaken for 5 min and read at 490 nm in a microplate scanning spectrophotometer [21]. The %
inhibition for each compound was calculated according to the following formula:
%
inhibition = (Activity of control – Activity of test compound)/ Activity of control × 100
The IC50 values were determined as the maximal inhibition by bliss method.
.3.3. In vitro COX-1/COX-2 inhibition assay
2
All the synthesized compounds were screened for their in-vitro COX-1 and COX-2 inhibitory
potential by using “COX inhibitory screening assay” kit (cayman chemical, Ann Arbor, MI)
followed by the manufacturer’s instruction [22]. Celecoxib was used as the standard for the
COX-2 and the indomethacin was used as the standard for the COX-1. Reaction mixture of tris-
HCl buffer (0.5M) pH 8.0, hematin (5mM), EDTA (0.5M), enzyme (COX-1 or COX-2) and the
tested compounds (different conc.) were made and pre-incubated at 25 °C for 5 min. then
colorimetric substrate and arachidonic acid was added to reaction mixture and was incubated for
further 20 min. Absorbance was measured at 420 nm using the plate reader [23]. The
concentrations of the tested compounds causing 50% inhibition (IC50) of the enzyme (COX-1 or
COX-2) were calculated by the dose response curve.
2
.3.4. In vivo Anti-inflammatory Activity
2
0

Anti-inflammatory activity was evaluated for the compounds showed good 5-LOX activity by
using Carrageenan-induced hind paw edema method [24]. Wistar rats (either sex) of weighing
1
75-220 g were procured from Central Animal House facility of Jamia Hamdard, New Delhi.
The experimental protocol was approved by the Institutional Animal Ethics Committee (IAEC)
Reg. No: 1287 CPCSEA] Jamia Hamdard and was performed according to CPCSEA guidelines.
[
During the experiment, Animals were kept under overnight fasting which were grouped into 19
groups (n = 3-6). Out of which one was standard group which received indomethacin (20 mg/kg
b.w) as the standard drug, one was control group to which only distilled water (1% CMC in
water in a volume of 10 ml/kg) was given and other were tested groups which was treated with
the tested compounds (20 mg/kg b.w). Tested compounds were given prior to 30 min of
carrageenan insertion to the tested group. 0.1ml of 0.1% of the Carrageenan (prepared in saline
water) was injected to the right hind paw (in sub-plantar region) of each rat to stimulate
inflammation. The volume of the paw edema was calculated by the digital Plethysmometer
(
PLM01) at 0 hour, 3 hour and 5 hour [25]. The average percent increase in paw volume with
time was calculated and compared against the control group. Percent inhibition of paw edema
was measure by using the formula:
V -Vt
c
%
inhibition =
×100
Vc
= mean increase in paw volume of control group
= mean increase in paw volume of test group
.3.5. Ulcerogenic activity
Where V
c
V
t
2
Compounds with good anti-inflammatory activity were further proceeding for ulcerogenic
activity [26]. Animals were divided into 4 groups; each group was having three animals.
Indomethacin (60mg/kg) was used as a standard. Tested compounds were administered with the
dose three times higher than the dose used for anti-inflammatory activity. After 5 hour of oral
administration, animals were scarified and stomach was removed and collected in 10% of the
formalin. The slides of the tissues were prepared by the standard method using hematoxylin and
eosin staining and were examined under light microscope.
2
1

2
.3.6. In vitro tubulin Assay
Compounds with excellent anti-proliferative activity were screened for tubuin assay [27] by
using tubulin polymerization assay kit from cytoskeleton, Inc. (BK011P) following the
manufacture’s instruction. The fluorescence polymerization assay was performed which was one
step procedure for determining the effects of drugs on tubulin polymerization. 2 mg/ ml of
Tubulin (derived from porcine brain) was added in 80 mM PIPES PH 6.9, 2.0 mM MgCl , 0.5
2
mM EGTA, 1.0 mM GTP, and 15% glycerol in presence or absence of test compounds to make
tubulin reaction mix. This reaction mix was poured into the 96 well plates and the plate reader
was started. Fluorescence emission was measured by the fluorimeter at 410-460nm (excitation
wavelength 340-360 nm). Paclitaxel and vincristine were used as the standard drug. Due to
presence of the fluorescence reporter on the microtubules, fluorescence enhancement was
measured by the fluorimeter when polymerization occurs.
2
.4. In silico Molecular docking studies
To view the binding pattern of synthesized compounds at molecular level, molecular docking
was done at COX-2 receptor catalytic ligand binding site (PDBID: 4RRX) [28], LOX-5 receptor
catalytic ligand binding site (PDB ID: 3V99) [29] and at ATP-binding site of Colchicine receptor
catalytic ligand binding site (PDB ID: 4O2B) [30]. The docking simulations of all the
compounds were performed using Maestro, version 9.6 implemented from Schrodinger software
suite. Synthesized compounds were sketched in 3D format using the build panel and were also
prepared for docking using lig prep application. The proteins were taken from Protein data bank
and were prepared for docking by solvent removing, hydrogen adding and further minimization
in the presence of bound ligand (COL) using protein preparation wizard. Grids for molecular
docking were generated with bound co-crystallized ligand. For validation of docking parameters
co-crystal ligand (COL) was re-docked at catalytic site of protein. RMSD between co-crystal and
re-docked pose was found to be 0.235 A [10]. Compounds were docked using Glide extra-
precision (XP) mode, with up to three poses saved per molecule.
3
. Result and Discussion
3
.1. Chemistry
2
2

A library of 1,2,3-triazole tethered indole-3-glyoxamide derivatives were synthesized by click
chemistry [31] approach as illustrated in scheme 1.
Scheme 1
Intermediate indole-3-glyoxalyl chloride 8 was obtained by treating indole 7 with oxalyl chloride
in ether at 0 °C. This indole-3-glyoxalyl chloride 8 was immediately reacted with propargyl
amine to afford indole-3-glyoxamide 9 with terminal alkyne. Propargylated intermediate 9 was
finally cyclized to triazole ring by reacting with different substituted of azides under Cu(I)
catalysed click chemistry conditions. A series of triazole derivatives of indole-3-glyoxamides
(
11a-11u) were synthesized by using different aromatic/sugar azides (10a-10u) by varying the
nature and site of substituents as depicted in Table 1. A.S. Hélio et al. have reported synthesis of
compounds 11c, 11d and 11h [32] but their spectral data and biological properties have not been
mentioned. However diisopropylethylamine (DIPEA) catalyzed amidation of indolglyolallyl
chloride with propargyl amine was resulted in the formation of 2-(1H-indol-3-yl)-2-oxo-N-(prop-
2
-ynyl)acetamide in 80%. In our case, pyridine catalysed amidation of indolglyolallyl chloride
with propargyl amine yielded the product 2-(1H-indol-3-yl)-2-oxo-N-(prop-2-ynyl)acetamide
9) in quantitative yields.
(
Propargylated intermediate 9 was treated with p-toluene sulphonyl chloride in dry pyridine to
yield 2-oxo-N-(prop-2-ynyl)-2-(1-tosyl-1H-indol-3-yl)acetamide (12) in quantitative amount.
When dry DCM was used instead of dry pyridine, the desired compound was obtained in low
yields (>5%).
N-tosylated glyoxamide derivative 12 with terminal alkyne was again treated with different
aromatic/sugar azides (10a-10u) to yield 1,2,3-triazole tethered N-tosyl indole-3-glyoxamide
derivatives 13a-13u as shown in Table 2.
1
13
All the synthesized compounds were characterized by H NMR, C NMR, IR, ESI-MS. In the
1
H NMR spectra, the resonance of H-C(5) of the triazollyl proton (δ 8.01-8.92 ppm) in the
aromatic region gave the evidence for the formation of triazoles. ESI-MS of all synthesized
+
+
+
compounds gave molecule fragement of [M] or [M + 1] or [M + 2] .
3
.2. Biological Evaluation
2
3

3
.2.1. Anti-proliferative Activity
All the synthesized compounds (11a-11u) and compounds (13a-13u) were evaluated for their
in-vitro anti-proliferative activity against selected human cancer cell lines of ovarian (SKOV3),
prostate (DU145), cervical (HELA) by using MTT assay. The IC50 values (µM) of the tested
compounds (11a-11u), (13a-13u) and the standard VP-16 are enlisted in Table 1 and Table 2.
From the obtained data it is evident that prostate (DU145) and cervical (HELA) cancer cell lines
were more susceptible for this class of compounds than ovarian (SKOV3) cell line. Most of the
compounds showed moderate to good anti-proliferative activity against DU145 and HELA
cancer cell lines. Compound 11g and 11q were demonstrated better growth inhibitions with IC50
values of 8.69 µM and 8.17 µM respectively against DU145 and found to be better than the
standard (VP16, IC50 9.8 µM).
Compound 11k displayed potent activity against cervical (HELA) cancer cell line with IC50 6.29
µM, where as the standard compound (VP16) showed IC50 of 7.43 µM. Compounds 11h, 11i,
1
1n, 11o, 11r also showed good to moderate cytotoxicity towards prostate (DU145) cancer cell
line (IC50 15.96 - 39.57 µM) and Compounds 11e, 11i, 11r illustrated good anti-proliferative
activity with IC50s ranging between 11.91 - 12.70 µM against ovarian (SKOV3) cancer cell line
as summarized in Table 1.
Table 1
In the next series of compounds (13a-13u), it was observed that N-tosylation on the indolyl
nitrogen lead to decrease the activity as compared to compounds without N-substitution (11a-
1
1u). Among this series, compound 13u was found to be the most potent compound against
cervical (HELA) cancer cell line with IC50 7.79 µM when compared with VP16 (IC50 7.43 µM).
Compounds 13b, 13l, 13r, 13s demonstrated moderate to good anti-proliferative activity against
DU145 cancer cell line with IC50s ranging between 11.88 - 18.53 µM as shown in Table 2.
Table 2
Dose response curve of compounds 11g and 11q against DU145 cell line are shown in fig. 3.
2
4

Figure 3
3
.2.2. In vitro COX-1, COX-2 and 5-LOX inhibitory activities
All the synthesized compounds (11a-11u) and (13a-13u) were evaluated for their in vitro COX-
, COX-2 and 5-LOX inhibitory potencies. Standard compounds indomethacin (COX-1
1
inhibitor) celecoxib (COX-2 inhibitor) and Nordihydroguaiaretic acid (5-LOX inhibitor) were
evaluated in parallel. Among compounds 11a-11u, compounds 11a, 11e, 11m and 11o
demonstrated moderate COX-1 inhibition with IC50s ranging between 2.02-2.03 µM when
compared with standard indomethacin (IC50 0.0068 µM) shown in Table 1. Whereas compounds
1
1d, 11l, 11m, 11q and 11r exhibited potent COX-2 inhibition with IC50s ranging between 0.12
0.19 µM (activity order: 11q > 11m > 11r > 11d > 11l) when compared to positive control
-
celecoxib (IC50 0.041 µM). Compounds 11b and 11d demonstrated potent 5-LOX inhibition
with IC50s 6.86 and 7.42 µM respectively and are comparable to standard NDGA (IC50 7.31
µM). Some of the other compounds also illustrated good 5-LOX inhibitory activity with order
1
1b > 11d > 11l=11r > 11m=11q (IC50s ranging between 6.85-7.73 µM) as summarized Table 1.
Compound 11g which was exhibiting excellent anti-proliferative activity against DU145
demonstrated the good 5-LOX inhibitory activity with IC50 8.62 µM. Based on the results
obtained from COX-1, COX-2 and 5-LOX inhibition studies, compound 11q with p-ethyl group
illustrated excellent COX-2/COX-1 selectivity index with 0.058.
From the synthesized compounds (13a-13u), Compound 13m (p-cyano group) showed a good
COX-1 inhibitory activity with IC50 2.13 µM. Whereas compound 13s having sugar moiety
(
glucosyl) instead of aromatic ring lead to the potent COX-2 inhibition (IC50 0.12 µM).
Compounds 13q and 13s demonstrated excellent 5-LOX inhibition with IC50s 7.43 and 7.46 µM
respectively and are comparable with the standard NDGA. Among all the synthesized molecules
of 13a-13u series, Compound 13s showed good COX-2 selectivity index of 0.046 as illustrated
Table 2. Dose response curve of compounds 11g, 11q and 13s against COX-1,COX-2, and 5-
LOX enzyme were shown in fig. 4, fig. 5 and fig. 6.
Figure 4
Figure 5
Figure 6
2
5

3
.2.3. In vivo anti-inflammatory activity
In vivo anti-inflammatory effect was evaluated for those compounds having good 5-LOX
inhibitory activity by using carragennan-induced hind paw edema model at equimolar
concentrations to the standard drug indomethacin (20mg/kg) and the results are depicted in Table
3
. Among the compounds tested, compounds 11b, 11l, 11q and 13q exhibited pronounced anti-
inflammatory activity as mentioned graphically in fig. 7 (69.2-76.2% inhibition at 3h, 75.0-
2.8% inhibition at 5h).
8
The anti-inflammatory activity profile of compounds 11q and 11b (76.90 & 76.92% inhibition
respectively at 3h post-carrageenan and 82.80 & 80.88% inhibition respectively at 5h post-
carrageenan) was found to be better than the standard NSAID, indomethacin (66.60% inhibition
at 3h as well as 77.94% inhibition at 5h) and showed a time-dependent increase in the inhibition
of inflammation. Compounds 11l and 13q demonstrated comparable anti-inflammatory activity
profiles to standard drug indomethacin.
Table 3
Figure 7
3
.2.4. Ulcerogenic studies
The compounds showing potential in vivo anti-inflammatory activity (11q and 13q) were further
carried out for gastric ulceration studies. Indomethacin was taken as the standard drug. Tested
compounds did not persuade any gastric ulceration and disruption of gastric epithelial cells. The
epithelial lining of the glandular cells are intact and no loss of epithelial cells were seen whereas
in standard compound lead to thinning and loss of the epithelial cell line as shown in fig. 8.
Figure 8
3
.2.5. Effect on tubulin polymerization
To investigate the mechanistic studies for the compound 11q, compound 11q was evaluated for
anti-tubulin activity by using the in vitro assay kit. Compound 11q (at 20µM) was compared
with paclitaxel (at 3µM) as microtubule stabilizing agents and vincristine as microtubule
2
6

destabilizing agents. The result showed that compound 11q interfered with the microtubule
dynamics. From the fig. 9, it was concluded compound 11q showed same dynamic curve as that
of vincristine. It lead to decrease the polymerization rate to some level which shrink microtubule
polymer cell mass in cell. so, 11q was act as microbule-destabilizing agents called as
polymerization inhibitor.
Figure 9
3
.3. Results of docking experiments
Using the Schrodinger software [33], docking studies were evaluated to find out the binding
mode of the active compound for its COX-2, 5-LOX and the tubulin inhibitory activity. Docking
score of the compounds is depicted in Table 4. The binding interaction of the active compounds
with respective of their activity can be justified by the in silico data.
Table 4
3
.3.1. Docking to COX-2 active centre
All the synthesized molecules were docked in the catalytic binding pocket of COX-2
receptor (PDB ID: 4RRX) which revealed a common binding orientation of the molecules.
The synthesized compounds (11a-11u and 13a-13u) were docked against COX-2 receptor
active site. The binding pose of the potent COX-2 Inhibitor 11q was observed and
compared with the Co-Crystal ligand (celecoxib). Compound 11q was interacted with the
backbone of COX-2 receptor with Tyr 385, Ser530 and Tyr355 amino acids and also
formed the hydrophobic interaction with Trp89, Phe381 and Ile354 amino acids as shown
in fig. 10. It was also superimposited with the Co-crystal ligand (celecoxib) which
displayed the same interaction in catalytic domain of COX-2 receptor as that of celecoxib.
Aromatic moiety attached to triazollyl ring of compound 11q is superimposing on the
aromatic moiety with sulphonamide functional group of celecoxib and triazollyl ring of
1
1q is slightly super imposing on pyrazole ring of celecoxib. Mostly para-substituted
alkyl and halogen substituted) compounds have shown similar docking results to that of
celecoxib due to hydrophobic interactions.
(
2
7

Figure 10
3
.3.2. Docking to 5-LOX active centre
Compound 11q exhibited a good COX-2 inhibitory activity and also good anti-prolifertive
activity which was further selected to docked in the catalytic binding pocket of 5-LOX receptor
(
PDB ID:3V99). Herein, Compound 11q formed the interaction with Gln557 amino acid of 5-
LOX receptor (in fig. 11) and also exhibited the hydrophobic bonding with Phe555 and Phe169.
Furtheron, it was superimposited with NDGA to see the positioning of the compound 11q in the
catalytic domain of the 5-LOX receptor. But NDGA (standard) demonstrated the higher docking
score than the synthesised compounds which revealed In silico data is irrespective to the
experimental data.
Figure 11
3
.3.3. Docking to Tubulin active centre
As most of the synthesized compounds showed excellent to good anti-proliferative activity.so
molecular docking of Compounds 11a-11u and 13a-13u were performed in the active binding
site of tubulin and were carried out to determine common binding orientation of all the
compounds in catalytic domain of ATP at colchicine binding site of tubulin (PDB ID : 4O2B).
The binding pose of most potent compounds 11q and 13s were explored and compared with the
standard Nocodazole as seen in fig. 8. Compound 11q showed a strong binding in the pocket of
tubulin with Lys352 and Val238 amino acids. Compound 11q was also involved in hydrophobic
bonding with Leu255 and Ile354 amino acid. Whereas compound 13s was interacted with the
Lys254, Asn249 and Gln247 residue in catylatic domain colchicine binding site. Compounds
1
1q and 13s were posed in superimposition with Nocodazole ligand as illustrated in fig. 12.
Furtheron, It was found that compound 11q exhibited similar kind of interaction as that of
nocodazole in catalytic domain of ATP at Colchicine binding site of tubulin. So the docking
score of 11q was higher than nocodazole and 13s showed lower docking score than nocodazole.
Figure 12
3
.4. Structure Activity Relationship
2
8

The structure activity relationship was summarized after overviewing the biological activity data.
Structure activity relationship of synthesized compounds has been depicted based on three
parameters: N-tosylation on indolyl nitrogen, nature of the group attached to 1,2,3-triazollyl ring,
nature of the substituents (functional group) and the position of substitutions on aromatic ring as
depicted in fig. 13.
The influence of N-tosylation on indolyl nitrogen (X = -SO
2
(C6H
5
)CH ) can be easily drawn
3
from the biological data as N-tosylated compounds (13a-13u) demonstrated reduced anti-
proliferative activity when compared to unsubstituted indole derivatives (X = H) (11a-11u). This
is also evidenced from the in silico molecular docking studies against tubulin. Indolyl –NH is
required for the key interactions with Val-238 at colchicine binding site of tubulin. With respect
to nature of the group attached to 1,2,3-triazollyl ring, aryl substitution on triazollyl ring (11g,
1
1i, 11k, 11q) showed superior anti-proliferative as well as dual COX-2 and 5-LOX inhibitory
activities over the glucosyl, galactosyl and pyridyl substitutions (11s, 11t, 11o, 11p).
Nature and position of the substituents had greatly influenced anti-proliferative as well as COX-2
and 5-LOX inhibitory activities. Electron donating groups and halogens substituted on aromatic
ring (11k, 11q and 11g) have shown promising anti-proliferative activity when compared to
electron withdrawing groups (other than fluorine). Most suited site for anti-proliferative activity
was found to be para position and 3,4-disubstitution on the phenyl ring. Anti-proliferative
activity with respect to the position of substitution on aromatic ring can be witten as
para>meta>ortho.
Compound with -fluoro substitution at para-position (11g) conferred significantly higher anti-
proliferative activity compared to chloro- (11c-e)/bromo- (11u) substituted compounds. Activity
profiles of halogen substituted compounds were observed as F > Cl > Br. Compound with p-
ethyl substitution (11q) on aromatic ring was found to be the best against DU145 cancer cell
lines and demonstrated better activity when compared to the standard drug etoposide (VP16).
Compound having 3,4 di-methyl substitution on phenyl ring (11k) displayed excellent anti-
proliferative activity when compared to the standard drug etoposide against HELA cancer cell
lines. It has also shown good 5-LOX inhibitory activity.
Compounds with both electron donating (11q, 11l, 13i, 13q) and electron withdrawing groups
(
11b, 11a) have demonstrated good anti-proliferative activity profiles. Substitution at para
position of phenyl ring retained the anti-inflammatory activity in both the series. From the in
2
9