Novel pyrrole derivatives bearing sulfonamide groups: Synthesis in?vitro cytotoxicity evaluation, molecular docking and DFT study

Article abstract of DOI:10.1016/j.molstruc.2017.06.003

The synthesis of new derivatives of pyrrole substituted sulfonamide groups is described. The in vitro anticancer activity of these pyrroles was evaluated against MCF7, MOLT-4 and HL-60 cells using MTT assay. The target compounds showed inhibitory activity against tested cell lines. Among the compounds, compound 1a exhibited good cytotoxic activity. The potential of this analog to induce apoptosis was confirmed in a nuclear morphological assay by Hoechst 33258 staining in the PC-12 cells. Finally, molecular docking was performed to determine the probable binding mode of the designed pyrrole derivatives into the active site of FGFR1 protein. DFT calculations were carried out at the B3LYP levels of theory with 6-31+G (d,p) basis set for compound 1a. The point group (C₁) of it was obtained based on the optimized structures; the calculation of the FT-IR vibrational frequencies, ¹H NMR and ¹³C NMR chemical shifts of the compound were carried out and compared with those obtained experimentally.

Full text of DOI:10.1016/j.molstruc.2017.06.003

Accepted Manuscript
Novel pyrrole derivatives bearing sulfonamide groups: Synthesis in vitro cytotoxicity
evaluation, molecular docking and DFT study
Masoumeh Bavadi, Khodabakhsh Niknam, Omolbanin Shahraki
PII:
S0022-2860(17)30778-0
10.1016/j.molstruc.2017.06.003
MOLSTR 23886
DOI:
Reference:
To appear in: Journal of Molecular Structure
Received Date: 1 March 2017
Revised Date: 4 May 2017
Accepted Date: 1 June 2017
Please cite this article as: M. Bavadi, K. Niknam, O. Shahraki, Novel pyrrole derivatives bearing
sulfonamide groups: Synthesis in vitro cytotoxicity evaluation, molecular docking and DFT study, Journal
of Molecular Structure (2017), doi: 10.1016/j.molstruc.2017.06.003.
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ACCEPTED MANUSCRIPT
Novel Pyrrole Derivatives bearing Sulfonamide Groups: Synthesis,
In Vitro Cytotoxicity Evaluation, Molecular Docking and DFT
Study
CO
2
2
Me
Me
O
MeO C
2
O
CO
2
MeO C
9
N
SO NHG
2
NH2
OH
OCH
3
HO
O
O
Ethanol
reflux
1 (a-d)
+
+
SO NHG
2
O
OCH
3
O
N
O
O
6(a-c)
7
8(a-c)
O
O
1
0
2
SO NHG
OCH
3
1
(e-i)

ACCEPTED MANUSCRIPT
Novel pyrrole derivatives bearing sulfonamide groups: Synthesis in vitro cytotoxicity
evaluation, molecular docking and DFT study
,
a
a
b
Masoumeh Bavadi, * Khodabakhsh Niknam and Omolbanin Shahraki,
a
Department of Chemistry, Faculty of Sciences, Persian Gulf University, Bushehr, 75169 Iran; E-mail:
mbavadi@gmail.com
b
Department of Medicinal Chemistry, Faculty of Pharmacy, Zahedan University of Medical Sciences,
Zahedan, Iran
Abstract
The synthesis of new derivatives of pyrrole substituted sulfonamide groups is described. The in vitro
anticancer activity of these pyrroles was evaluated against MCF7, MOLT-4 and HL-60 cells using MTT
assay. The target compounds showed inhibitory activity against tested cell lines. Among the compounds,
compound 1a exhibited good cytotoxic activity. The potential of this analog to induce apoptosis was
confirmed in a nuclear morphological assay by Hoechst 33258 staining in the PC-12 cells. Finally,
molecular docking was performed to determine the probable binding mode of the designed pyrrole
derivatives into the active site of FGFR1 protein. DFT calculations were carried out at the B3LYP levels of
theory with 6-31+G (d,p) basis set for compound 1a. The point group (C ) of it was obtained based on the
1
1
13
optimized structures; the calculation of the FT-IR vibrational frequencies, H NMR and C NMR chemical
shifts of the compound were carried out and compared with those obtained experimentally.
Keywords: Pyrrole, Anticancer, Sulfonamide, MTT, Molecular docking, DFT
1
.
Introduction
These days the number of cancer related mortality is increasing and one of the most leading causes of death
in the world is cancer. Despite the immense advances and efforts in the field of cancer research, no
currently available drugs can exterminate cancer cells without harming normal tissues [1]. The fibroblast
growth factor (FGF) that signals through FGF receptors (FGFR1, FGFR2, FGFR3 and FGFR4) plays an
important role in many physiologic processes, comprising embryogenesis, regulation of angiogenesis,
wound healing and inflammation [2-4]. These families of proteins are expressed on various types of cells
and regulate cellular actions, such as proliferation, differentiation, survival and metastasis of tumor cells. In
this view, they are considered as the targets for cancer therapy [5].
There are several reports on small molecule FGFR inhibitors. Some of them such as NVP-BGJ398
and AZD4547 have entered clinical trials and LY2874455 showed good FGFR inhibitory activity [6] (Fig.
1
). Different series of nitrogen containing five-membered heterocyclic compounds have been reported as
inhibitors of FGFR [7-9]. Therefore, the development and discovery of new anticancer agents with
promising activity, selective and high therapeutic index is still a major challenge to medicinal chemists.
1

ACCEPTED MANUSCRIPT
O
HN
Cl
N
O
O
O
N
N
N
H
N
O
N
Cl
O
N
NH
N
N
H
AZD4547
NVP-BGJ398
H
N
Cl
N
O
N
Cl
LY2874455
Fig. 1. The structure of some FGFR inhibitors.
N
N
OH
Pyrrole and its derivatives are well known as the most important nitrogen-containing heterocyclic
compounds due to their present as structural subunits of many natural products including heme, vitamin
B12, and cytochromes, bile pigments, and alkaloids [10, 11].
F
R1
R
CO H
2
HO C
O
N
2
N
O
Me
HO
HO
HN
CH3
H C
R= H, R1= Me
Tolmetin
3
Atorvastatin
Me N
N
N
2
N
Me
N
Me
Me
Cl
N
pyrvinium
Fig. 2. The biologically active compounds containing the pyrrole moiety.
BM212
Cl
They have also exhibited remarkable biological activities such as antibacterial [12], antitubercular [13],
antitumor [14], anti-inflammatory [15] and antioxidant activity [16]. Some biologically important
compounds containing the pyrrole moiety such as atorvastatin (a cholesterol-lowering agent), tolmetin and
anthelmintic pyrvinium (nonsteroidal anti-inflammatory drugs), BM212 and its derivatives (Mycobacterium
2

ACCEPTED MANUSCRIPT
tuberculosis activity) [17-21] are shown in Fig. 2. Moreover, pyrroles are used as insecticides [22] and
useful building units in the field of material chemistry [23].
In recent years, the synthesis of drugs bearing sulfonamide groups in their structures has been
attended. They are exhibited a significant biological properties including anticancer [24], antimalarial [25],
anti-inflammatory [26], antihypertensive [27], anticonvulsant, and herbicidal properties [28]. A series of
these drugs such as sulfonamide KCN1 which was reported as an antitumor agent and the HIV protease
inhibitor amprenavir was used for the treatment of AIDS and HIV infections [29, 30]. In addition, it was
reported that sulfonamides were used as inhibitors of the activity of the enzymes for example;
dihydropteroate synthase (DHPS) [31], matrix metalloproteinase [32], and carbonic anhydrase (CA) [33].
H
SO NH
2
2
R1
N
R2
R3
O
O
S
S
O
Cl
N
N
O
O
H
H
NH
benzoxazine-6-sulfonamides
E7070
O
S
H N
2
O
N N
S
S
N
O
O
N
N
S NH2
O
N
O
H
AZA
MZA
Fig. 3. Chemical structures of some therapeutic sulfonamides.
Moreover, sulfonamide drugs have been reported such as benzoxazine-6-sulfonamides and E7070
(
[
anticancer agents) [34, 35], and anti-glaucoma compounds acetazolamide AZA and methazolamide MZA
36-38] (Fig. 3). Therefore, due to the pharmacological and biological importance of sulfonamides and
nitrogen-containing heterocyclic compounds in medicinal chemistry, the synthesis of compounds bearing
sulfonamide moieties with the aim of outstanding biological properties has become interesting field in
research.
In this study, we reported the synthesis of a new class of pyrrole derivatives substituted
sulfonamide groups. These compounds were subsequently evaluated for anticancer activity and
3

ACCEPTED MANUSCRIPT
morphological study. The binding mode of these derivatives with the active site of the FGFR1 protein was
also investigated by molecular docking.
2
.
Experimental Section
2
.1. Materials and methods
The chemicals were purchased from Merck and Aldrich Chemical Companies. The progress of the reactions
1
13
was monitored by TLC on silica gel PolyGram SILG/UV254 plates. For recorded HNMR and C NMR
400 MHZ and 100 MHZ, respectively) spectra we used Bruker (400 MHz) Avance Ultrashield in pure
deuterated DMSO-d solvent with tetramethylsilane (TMS) as internal standards. The FT-IR spectroscopy
(
6
(
(
Shimadzu FT-IR 8300 spectrophotometer) was run for characterization of the products. Mass spectra
FINNIGAN-MAT 8430 mass spectrometer) were employed at 70 eV. Melting points were obtained in
open capillary tubes in a Barnstead Electrothermal 9100 BZ circulating oil melting point apparatus. PC-12
rat pheochromocytoma), MCF7 (human breast adenocarcinoma), MOLT-4 (human acute lymphoblastic
(
leukemia) and HL-60 (promyelocytic leukemia) cells were obtained from the National Cell Bank of Iran
Pasteur Institute (Tehran, Iran). The cell culture medium (RPMI 1640), fetal bovine serum (FBS), and
penicillin–streptomycin were obtained from Gibco BRL (Life technology, Paisley, Scotland). 3-(4,5-
Dimethyltiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) powder and Hoechst 33258 were purchased
from Sigma Chemicals Co. (Germany). The culture plates were purchased from Nunc (Denmark).
2
.2 Cytotoxicity activities
2
.2.1. Cell culture
All cell lines were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 µg/mL
o
streptomycin, and 100 U/mL penicillin at 37 C in a humidified incubator with 5% CO .
2
2
.2.2.
Cytotoxicity assay
The cytotoxicity activity of the synthesized compounds was determined by 3-(4,5-dimethylthiazol-2-yl)-
4
2
,5-diphenyl-tetrazolium bromide (MTT) assay. The cells were seeded in 96-well plates (5 × 10 cells/mL)
and incubated for 24 h to allow cell attachment. In order to prepare stock solution, all of the synthesized
compounds were dissolved in DMSO. The final concentration of DMSO in the culture medium did not
exceed 0.5%. Different concentrations of the compounds (25, 50, 100 and 200 µM) were prepared in
4

ACCEPTED MANUSCRIPT
culture medium and added to the wells in triplicate and incubated for another 72 h. In the following, 800 µL
of MTT reagent (5 mg/mL) was diluted in 5.6 mL phosphate buffered serum (PBS). 80 µL of the obtained
o
MTT solution was added to each well and the plates incubated for an additional 3 h at 37 C. Thereafter, the
culture medium was removed and the formazan precipitates were dissolved in 200 µL dimethylsulfoxide
(
DMSO). Cisplatin was used as a positive control. Finally, the absorbance of each well was determined by
plate reader (Anthous 2020; Austria) at 570 nm with background correction at 655 nm. Cell viability was
calculated using the following formula:
Where Ab (S), Ab (B) and Ab (C) are the colorimetric intensity of the cells incubated with the samples,
blank (wells contained only growth medium for background correction) and negative control (wells
contained cells but no drugs) respectively. IC50 values were calculated with the software CurveExpert
version 1.34 for Windows.
2
.2.3. Morphological study
3
PC-12 cells (5 ×10 /well) were seeded in 24-well plates and incubated for 24 h. The cells were treated with
and without compound 1a for 48 h. After incubation, the cell morphology was assessed by Axiovert 25
phase-contrast microscope (Zeiss, Germany).
2
.2.4. Hoechst 33258 staining Assay
The ability of the target compounds to induce apoptosis in PC-12 cells was evaluated morphologically by
4
Hoechst 33258 staining test [39]. Briefly, the cells (7 × 10 /well) were seeded in 12-well plates and
o
incubated in a humidified atmosphere at 37 C with 5% CO . Then, the cells were treated with and without
2
compound 1a for 48 h. Plates were washed with PBS and stained with 0.5 µL of Hoechst 33258 (10
o
mg/mL) and incubated at 37 C for 15 min at room temperature. Afterwards, the cells were immediately
analyzed by an Axoscope2 plus fluorescence microscope from Zeiss (Germany).
2
.3. Molecular docking
The crystal structure of FGFR1 was obtained from the Protein Data Bank (PDB code: 4ZSA). Water
molecules and co-crystalized ligands were excluded. Validation of molecular docking protocol was checked
via testing the ability of AUTODOCK 4.2 to reproduce the binding mode of cognate ligand. The PDB file
5

ACCEPTED MANUSCRIPT
of protein was regenerated. An N-terminus of a chain is capped with ACE (acetyl) while C-terminus is
capped with NME (formyl) moiety. The Reduce program [40] used to determine the hydrogen coordinates
using steric considerations and geometric hydrogen bond network analysis; ionization and flipped states of
His, Glu and Asp are assigned on geometric grounds alone and rotamers are assigned from a fixed discrete
collection. The loops were modeled by MODELLER 9v2 program [41] by merging the appropriate residues
in missing loop locations. The three-dimensional structures of compounds were built using the Avogadro
software. Structure optimization at [B3LYP combined to 6-31G (d,p) and 6-31+G(d,p)] level of theory was
performed using Gaussian 09 package software. The degrees of torsions were defined to generate PDBQT
format of the ligands. The grid parameter file was created using AutoGrid implemented in the MGLTools
package. The grid size was set to 60 × 60 × 60 points with spacing value 0.375 A. The prepared ligands
were docked in the rigid macromolecule using Lamarckian Genetic Algorithm (LGA), with 200 GA runs
and 25000 000 energy evaluations while the other parameters were left as default.
Molecular docking was performed by AutoDock 4.2 and results were analyzed with the aid of the
AutoDockTools-1.5.6. Cluster analysis was done on the docking results using a Root mean square deviation
(
RMSDꢀ≤ꢀ2 Å).
2
.4. General procedure for the synthesis of pyrrole derivatives bearing sulfonamide groups (1)
A mixture of sulfonamide (6; 1 mmol), phenylglyoxal monohydrate (7; 1 mmol), 1,3-dicarbonyl compound (8;
mmol), dimethyl acetylene dicarboxylate (9; 1 mmol) or acetylacetone (10; 1 mmol) in ethanol (96%, 5 mL)
1
was refluxed with stirring in an oil bath. The progress of the reaction was monitored by TLC. After completion
of the reaction, the resulting solids were filtered and washed with warm ethanol to obtain pure product [42].
Dimethyl-4-(1,3-dimethyl-2,4,6-trioxohexahydropyrimidin-5-yl)-1-(4-methoxy-3-
(
morpholinosulfonyl)phenyl)-5-phenyl-1H-pyrrole-2,3-dicarboxylate (1a): Light pink solid; Yield:
o
–1
9
1
3
7
5%; m.p. 177-179 C; IR (KBr) (ν ,cm ): 3000, 2980, 2850 (C–H), 1522 (C=C), 1729, 1678 (C=O),
max
1
344, 1165 (SO ); H NMR (400 MHz, DMSO-d ) δ (ppm): 2.78 (m, 4H, 2 × CH ), 3.18 (s, 6H, 2 × CH ),
2
6
2
3
.52–3.54 (m, 4H, 2 × CH ), 3.64 (s, 3H, CH ), 3.69 (s, 3H, CH ), 3.90 (s, 3H, OCH ), 4.98 (s, 1H, CH),
2
3
3
3
13
.20-7.22 (m, 2H, Ar), 7.30-7.36 (m, 5H, Ar), 7.68 (dd, 1H, J = 8.8, J = 2.8 Hz, Ar); C NMR (100 MHz,
1
2
DMSO-d ) δ (ppm): 28.9, 46.1, 52.3, 53.3, 56.9, 66.1, 114.3, 116.0, 124.7, 128.4, 128.8, 128.9, 129.2,
6
1
29.3, 129.7, 130.7, 131.1, 134.8, 138.7, 152.1, 157.0, 162.0, 163.9, and 168.0; Mass spectrum m/z: 668.3
+
[
M ]; Anal. Calcd. (%) C H N O S: C, 55.68; H, 4.82; N, 8.38; S, 4.80; Found: C, 55.37; H, 4.79, N,
3
1
32
4
11
8
.11; S, 4.48.
Dimethyl-4-(1,3-dimethyl-2,4,6-trioxohexahydropyrimidin-5-yl)-1-(4-methoxy-3-(N-
phenylsulfamoyl)phenyl)-5-phenyl-1H-pyrrole-2,3-dicarboxylate (1b): Light pink solid; Yield: 96%,
6

ACCEPTED MANUSCRIPT
o
–1
m.p. 207-209 C; IR (KBr) (ν ,cm ): 3288 (N–H), 3100 (C–H), 1495 (C=C), 1723, 1692 (C=O), 1332,
max
1
1
3
158 (SO ); H NMR (400 MHz, DMSO-d ) δ (ppm): 3.18 (s, 6H, 2 × CH ), 3.56 (s, 3H, OCH ), 3.63 (s,
2
6
3
3
H, OCH ), 3.89 (s, 3H, OCH ), 4.97 (s, 1H, CH), 6.99-7.05 (m, 3H, Ar), 7.11-7.14 (m, 3H, Ar), 7.20-7.28
3
3
(
m, 4H, Ar), 7.32-7.36 (m, 1H, Ar), 7.42 (dd, 1H, J = 8.8, J = 2.8 Hz, Ar), 7.57 (d, 1H, J = 2.8 Hz, Ar),
1 2
1
3
1
1
1
0.20 (s, 1H, SO NH); C NMR (100 MHz, DMSO-d ) δ (ppm): 28.9, 48.2, 52.3, 53.2, 57.1, 113.2, 113.8,
2 6
15.9, 119.8, 124.2, 126.8, 128.5, 128.7, 129.1, 129.4, 129.5, 129.9, 130.0, 130.3, 130.9, 134.8, 137.8,
+
38.6, 152.2, 156.6, 161.9, 163.8, and 168.0; Mass spectrum m/z: 674.3 [M ]; Anal. Calcd. (%)
C H N O S: C, 58.75; H, 4.48; N, 8.30; S, 4.75; Found: C, 58.43; H, 4.51, N, 7.98; S, 4.46.
3
3
30
4
10
Dimethyl-1-(3-(N-(chlorophenyl)sulfamoyl)-4-methoxyphenyl)-4-(1,3-dimethyl-2,4,6-
trioxohexahydropyrimidin-5-yl)-5-phenyl-1H-pyrrole-2,3-dicarboxylate (1c): Light pink solid; Yield:
o
–1
9
1
3%; m.p. ˃ 250 C; IR (KBr) (ν ,cm ): 3288 (N–H), 3050 (C–H), 1517 (C=C), 1722, 1691 (C=O),
max
1
322, 1160 (SO ); H NMR (400 MHz, DMSO-d ) δ (ppm): 3.18 (s, 6H, 2 × CH ), 3.59 (s, 3H, CH ), 3.63
2
6
3
3
(
s, 3H, CH ), 3.89 (s, 3H, OCH ), 4.97 (s, 1H, CH), 7.01-7.04 (m, 1H, Ar), 7.11-7.14 (m, 3H, Ar), 7.24-7.33
3 3
(
m, 6H, Ar), 7.44 (dd, 1H, J = 8.8, J = 2.8 Hz, Ar), 7.56 (d, 1H, J = 2.8 Hz, Ar), 10.36 (s, 1H, SO NH);
1 2 2
1
3
C NMR (100 MHz, DMSO-d ) δ (ppm): 28.9, 48.2, 52.3, 53.2, 57.1, 113.8, 115.9, 121.4, 126.2, 128.3,
6
1
28.5, 128.7, 129.0, 129.3, 129.5, 129.9, 130.0, 130.9, 135.0, 136.8, 138.6, 152.2, 156.6, 161.9, 163.8, and
+2
+
1
68.0; Mass spectrum m/z: 710.3 [M ], 708.3 [M ]; Anal. Calcd. (%) C H ClN O S: C, 55.89; H, 4.12;
33 29 4 10
Cl, 5.00; N, 7.90; S, 4.52; Found: C, 55.61; H, 4.15, N, 7.57; S, 4.18.
Dimethyl-1-(3-(N-(4-chlorophenyl)sulfamoyl)-4-methoxyphenyl)-5-phenyl-4-(2,4,6-
trioxohexahydropyrimidin-5-yl)-1H-pyrrole-2,3-dicarboxlate (1d): White solid; Yield: 88%; m.p. ˃ 250
o
–1
C; IR (KBr) (νmax,cm ): 3241, 3184 (N–H), 3075 (C–H), 1538 (C=C), 1750, 1710 (C=O), 1342, 1161
1
(
SO ); H NMR (400 MHz, DMSO-d ) δ (ppm): 3.59 (s, 3H, CH ), 3.67 (s, 3H, CH ), 3.89 (s, 3H, OCH ),
2
6
3
3
3
4
.82 (s, 1H, CH), 6.91–6.93 (m, 1H, Ar), 7.01–7.03 (m, 1H, Ar), 7.10–7.14 (m, 3H, Ar), 7.22–7.35 (m, 5H,
Ar), 7.42 (dd, 1H, J = 9.0, J = 2.4 Hz, Ar), 7.54 (d, 1H, J = 2.4 Hz, Ar), 10.36 (s, 1H, SO NH), 11.31 (s,
1
2
2
1
3
2
1
1
H, NH); C NMR (100 MHz, DMSO-d ) δ (ppm): 48.1, 52.1, 53.2, 57.1, 113.8, 115.6, 121.4, 126.1,
6
28.2, 128.3, 128.5, 128.8, 128.9, 129.2, 129.4, 129.5, 129.9, 130.0, 130.1, 130.3, 130.9, 135.0, 136.8,
+2
+
51.4, 156.6, 162.0, 163.8, and 169.6; Mass spectrum m/z: 682.7 [M ], 680.9 [M ]; Anal. Calcd. (%)
C H ClN O S: C, 54.67; H, 3.70; Cl, 5.21; N, 8.23; S, 4.71; Found: C, 54.34; H, 3.64, N, 7.91; S, 4.43.
3
1
25
4
10
5
-(3-Acetyl-4-(1,3-dimethyl-2,4,6-trioxohexahydropyrimidin-5-yl)-2-methyl-5-phenyl-1H-pyrrol-1-
o
yl)-2-methoxy-N-phenyl-benzenesulfonamide (1e): White solid; Yield: 94%; m.p. ˃ 250 C; IR (KBr)
–
1
1
(
νmax,cm ): 3419 (O–H, N–H), 3072 (C–H), 1573 (C=C), 1670 (C=O), 1346, 1159 (SO ); H NMR (400
2
MHz, DMSO-d ) δ (ppm): 2.13 (s, 3H, CH ), 2.16 (s, 3H, CH ), 3.05 (s, 6H, CH ), 3.89 (s, 3H, OCH ),
6
3
3
3
3
6
1
5
1
.95-6.99 (m, 5H, Ar), 7.00-7.03 (m, 3H, Ar), 7.14 (d, 1H, J = 8.8 Hz, Ar), 7.19-7.25 (m, 3H, Ar), 7.44 (d,
13
H, J = 2.8 Hz, Ar), 10.16 (s, 1H, SO NH); C NMR (100 MHz, DMSO-d ) δ (ppm): 13.5, 27.6, 29.3,
2
6
7.0, 82.2, 113.8, 119.7, 120.1, 123.7, 124.1, 126.2, 126.5, 127.6, 129.5, 130.2, 130.5, 130.5, 132.8, 133.1,
+
34.9, 135.1, 138.0, 153.6, 155.8, 162.7, and 196.8; Mass spectrum m/z: 614.4 [M ]; Anal. Calcd. (%)
C H N O S: C, 62.53; H, 4.92; N, 9.11; S, 5.22; Found: C, 62.24; H, 4.88, N, 8.82; S, 4.89.
3
2
30
4
7
1
H NMR (400 MHz, DMSO-d + D O) δ (ppm): 2.11 (s, 3H, CH ), 2.15 (s, 3H, CH ), 3.05 (s, 6H, CH ),
6
2
3
3
3
3
.87 (s, 3H, OCH ), 6.95-6.96 (m, 4H, Ar), 6.98-7.04 (m, 4H, Ar), 7.12 (d, 1H, J = 8.8 Hz, Ar), 7.19-7.28
3
(
m, 3H, Ar), 7.45(d, 1H, J = 2.4 Hz, Ar).
7

ACCEPTED MANUSCRIPT
5
-(3-Acetyl-4-(1,3-dimethyl-2,4,6-trioxohexahydropyrimidin-5-yl)-2-methyl-5-phenyl-1H-pyrrol-1yl)-
o
N-(4-chlorophenyl)-2-methoxybenzenesulfonamide (1f): White solid; Yield: 89%; m.p. ˃ 250 C; IR
–
1
1
(
KBr) (νmax,cm ): 3419 (O–H, N–H), 3070 (C–H), 1574 (C=C), 1667, 1646 (C=O), 1350, 1160 (SO ); H
2
NMR (400 MHz, DMSO-d ) δ (ppm): 2.15 (s, 3H, CH ), 2.16 (s, 3H, CH ), 3.05 (s, 6H, CH ), 3.89 (s, 3H,
6
3
3
3
OCH ), 6.95-6.99 (m, 5H, Ar), 7.01-7.04 (m, 2H, Ar), 7.15 (d, 1H, J = 8.8 Hz, Ar), 7.25-7.29 (m, 3H, Ar),
3
1
3
7
2
1
.42 (d, 1H, J = 2.4 Hz, Ar), 10.33 (brs, 1H, SO NH); C NMR (100 MHz, DMSO-d ) δ (ppm): 13.5, 27.6,
2 6
9.3, 57.0, 82.2, 113.8, 120.1, 121.4, 123.7, 126.2, 127.6, 128.0, 129.4, 130.2, 130.5, 130.5, 132.8, 133.1,
+2
+
34.9, 135.2, 153.6, 155.8, 162.7, and 196.8; Mass spectrum m/z: 650.3 [M ], 648.3 [M ]; Anal. Calcd.
(
%) C H ClN O S: C, 59.21; H, 4.50; Cl, 5.46; N, 8.63; S, 4.94; Found: C, 58.87; H, 4.45, N, 8.32; S,
32 29 4 7
4
5
.63.
-(3-Acetyl-4-(4-hydroxy-2-oxo-2H-chromen-3-yl)-2-methyl-5-phenyl-1H-pyrrol-1-yl)-2-methoxy-N-
o
–1
phenylbenzenesulfonamide (1g): White solid; Yield: 83%; m.p. 219–221 C; IR (KBr) (ν ,cm ): 3419
max
1
(
O–H, N–H), 3080 (C–H), 1495 (C=C), 1635, 1600 (C=O), 1342, 1158 (SO ); H NMR (400 MHz, DMSO-
2
d ) δ (ppm): 2.16 (s, 3H, CH ), 2.17 (s, 3H, CH ), 3.89 (s, 3H, OCH ), 6.96 (m, 5H, Ar), 7.01-7.04 (m, 3H,
6
3
3
3
Ar), 7.09-7.16 (m, 3H, Ar), 7.19-7.23 (m, 2H, Ar), 7.33-7.37 (m, 2H, Ar), 7.49 (s, 1H, Ar), 7.80 (d, 1H, J =
1
3
7
1
1
6
3
.2 Hz, Ar), 10.18 (s, 1H, SO NH); C NMR (100 MHz, DMSO-d ) δ (ppm): 13.5, 29.3, 57.0, 93.6, 113.9,
2 6
15.8, 119.7, 121.9, 124.1, 125.3, 126.4, 126.5, 127.7, 129.1, 129.5, 130.0, 130.1, 130.3, 130.5, 135.1,
+
38.0, 154.3, 155.9, 164.4, and 196.5; Mass spectrum m/z: 620.7 [M ]; Anal. Calcd. (%) C H N O S: C,
35
28
2
7
7.73; H, 4.55; N, 4.51; S, 5.17; Found: C, 67.41; H, 4.52, N, 4.20; S, 4.87.
-(4-Acetyl-1-(4-methoxy-3-(morpholinosulfonyl)phenyl)-5-methyl-2-phenyl-1H-pyrrol-3-yl)-4-
o
–1
hydroxy-2H-chromen-2-one (1h): White solid; Yield: 85%; m.p. 245–247 C; IR (KBr) (ν ,cm ): 3419
max
1
(
O–H), 3080 (C–H), 1496 (C=C), 1638, 1600 (C=O), 1343, 1161 (SO ); H NMR (400 MHz, DMSO-d ) δ
2
6
(
ppm): 2.17 (s, 3H, CH ), 2.37 (s, 3H, CH ), 2.68–2.71 (m, 4H, 2 × CH ), 3.53 (m, 4H, 2 × CH ), 3.92 (s,
3 3 2 2
13
3
H, OCH ), 6.98–7.15 (m, 8H, Ar), 7.33–7.40 (m, 2H, Ar), 7.73–7.89 (m, 2H, Ar); C NMR (100 MHz,
3
DMSO-d ) δ (ppm): 13.7, 29.3, 46.0, 56.9, 66.1, 93.3, 114.2, 115.8, 120.1, 122.0, 123.5, 124.6, 125.4,
6
1
26.3, 127.8, 130.0, 130.4, 130.6, 131.4, 132.5, 133.3, 135.1, 135.3, 154.3, 156.2, and 196.5; Mass
+
spectrum m/z: 614.6 [M ]; Anal. Calcd. (%) C H N O S: C, 64.48; H, 4.92; N, 4.56; S, 5.22; Found: C,
33
30
2
8
6
4.16; H, 4.87, N, 4.38; S, 4.91.
5
-(3-Acetyl-2-methyl-5-phenyl-4-(2,4,6-trioxohexahydro-pyrimidin-5-yl)-1H-pyrrol-1-yl)-2-methoxy-
o
–1
N-phenyl-benzenesulfonamide (1i): White solid; Yield: 91%, m.p. ˃ 250 C; IR (KBr) (ν ,cm ): 3395,
max
1
3
252 (N–H), 3100 (C–H), 1498 (C=C), 1725, 1634 (C=O), 1344, 1161 (SO ); H NMR (400 MHz, DMSO-
2
d ) δ (ppm): 2.25 (s, 3H, CH ), 2.40 (s, 3H, CH ), 3.91 (s, 3H, OCH ), 4.49 (s, 1H, CH), 7.00-7.04 (m, 5H,
6
3
3
3
Ar), 7.16-7.25 (m, 6H, Ar), 7.40 (dd, 1H, J = 8.8, J = 2.8 Hz, Ar), 7.51 (d, 1H, J = 2.8 Hz, Ar), 10.21 (s,
1
2
1
3
1
1
1
6
H, SO NH), 11.05 (s, 2H, NH); C NMR (100 MHz, DMSO-d ) δ (ppm): 14.1, 30.7, 48.5, 57.1, 114.0,
2 6
14.7, 119.6, 124.1, 126.6, 128.4, 128.8, 129.3, 129.5, 130.0, 130.2, 130.6, 131.0, 135.6, 136.6, 137.0,
+
38.0, 151.8, 156.4, 169.8, and 194.6; Mass spectrum m/z: 586.6 [M ]; Anal. Calcd. (%) C H N O S: C,
30
26
4
7
1.42; H, 4.47; N, 9.55; S, 5.47; Found: C, 61.11; H, 4.50, N, 4.32; S, 5.19.
3
.
Results and Discussion
3
.1. Chemistry
8

ACCEPTED MANUSCRIPT
The synthetic route to obtain pyrrole compounds bearing sulfonamide groups (1a-i) is outlined in
Scheme 1. Initially, sulfonamide derivatives (6a-c) have been carried out according to the reported
procedure [43, 44].
NH2
NHCOCH
3
NH2
a) R = H; R' = C H -
6
5
1
) (CH CO) O
1) RR'NH
2) HCl, H O
3
2
b) R = H; R' = 4-Cl-C H -
6 4
2
) HSO Cl
3
SO Cl
2
SO NRR'
2
2
C)
O
NH
OCH3
OCH
3
OCH3
2
4
6
CO Me
2
O
MeO C
2
O
CO Me
2
MeO C
2
9
N
SO NRR'
2
NH2
OH
OCH
3
HO
O
O
Ethanol
reflux
1 (a-d)
+
+
SO NRR'
2
O
OCH3
O
N
O
O
6(a-c)
7
8(a-c)
O
O
10
O
O
O
OH
H
H
H C
CH3
O
SO NRR'
2
3
N
N
N
N
=
OCH3
O
O
O
O
O
O
1
(e-i)
8a
8b
8c
Scheme 1 Synthesis of pyrrole derivatives bearing sulfonamide groups 1a-i
As demonstrated in Scheme 1, N-(4-methoxyphenyl)acetamide 3 was obtained via the reaction
between p-anisidine 2 and acetic anhydride which was treated with chlorosulfonic acid to give the
intermediate 4. The arylsulfonyl chloride intermediate 4 was converted into corresponding sulfonamide 5 in
the presence of various amines and anhydrous NaHCO at room temperature and under solvent-free
3
conditions. Finally, the compounds (5a-c) were hydrolyzed with water and acid to give sulfonamide
derivatives (6a-c) in high yield and purity. The reaction of sulfonamide derivatives (6a-c) with
acetylacetone or dimethyl acetylene dicarboxylate, 1, 3-dicarbonyl compounds and phenylglyoxal
monohydrate under catalyst-free conditions afforded corresponding final compounds 1a-i (Scheme 2).
9

ACCEPTED MANUSCRIPT
H C
H
3
C
N
O
H C
3
3
O
O
N
N
O
O
N
O
N CH3
O
CH3
N CH3
O
H CO C
H
3
CO
2
C
H
3
CO C
2
3
2
O
H CO C
H CO
3 2
C
H
3
CO
2
C
3
2
N
N
N
S
N
O
SO
2
NH
SO
2
NH
Cl
1a
O
1b
1c
2
OCH3
OCH
3
OCH
3
Time = 30 min; Yield = 95%
Amine = 6c and Dicarbonyl 8b
Time = 30 min; Yield = 96%
Amine = 6a and Dicarbonyl 8b
Time = 30 min; Yield = 93%
Amine = 6b and Dicarbonyl 8b
H
O
H C
3
O
H C
3
O
N
N
N
HO
O
HO
N H
O
N CH3
O
O
N
CH3
H CO C
3
2
O
O
H CO C
3
2
N
N
N
SO NH
Cl
SO NH
2
2
SO NH
Cl
1d
1e
2
OCH3
OCH3
1f
OCH3
Time = 120 min; Yield = 88%
Amine = 6b and Dicarbonyl 8a
Time = 30 min; Yield = 94%
Amine = 6a and Dicarbonyl 8b
Time = 30 min; Yield = 89%
Amine = 6b and Dicarbonyl 8b
H
O
O
O
N
O
HO
O
HO
O
N
O
H
O
O
O
N
N
N
SO NH
2
S N
O
SO NH
2
1
g
1h
1i
O
OCH3
2
OCH3
OCH3
Time = 120 min; Yield = 83%
Amine = 6a and Dicarbonyl 8c
Time = 120 min; Yield = 85%
Amine = 6c and Dicarbonyl 8c
Time = 50 min; Yield = 91%
Amine = 6a and Dicarbonyl 8a
Scheme 2 The structure of synthesized pyrrole derivatives bearing sulfonamide groups 1a-i
High chemical yields were achieved in short reaction time and without the need for column
chromatography or recrystallization. It is worth mentioning, there are possible tautomeric structures
(
Scheme 3) for the synthesized pyrroles. These tautomeric structures come from common hydrogen
bonding interaction between hydrogen and oxygen or nitrogen [45, 46]. The structures of these pyrrole
derivatives were elucidated on the running spectral data and elemental analysis. The IR spectra of
synthesized pyrroles 1a-i approved clearly the certain structures by observing the absorption bands for
1
C=O, NH, and SO functions at their determined regions. The H NMR spectra of pyrroles 1a-c were shown
2
the singlet peaks at 3.18 ppm and in the region of 3.56-3.90 ppm for the methyl and methoxy protons
1
0

ACCEPTED MANUSCRIPT
respectively. The CH proton of the pyrimidine ring was appeared as singlet between 4.97 and 4.98 ppm.
Also, the aromatic protons and the NH proton of the sulfonamide moiety were observed in the region of
1
3
6
.99-7.68 ppm and 10.21-10.36 ppm, respectively. The C NMR spectra of these pyrroles were appeared
signals between 152.1 and 168.0 ppm corresponding to the carbonyl carbon atoms and signals between
13.2 and 138.6 ppm corresponding to the carbon–carbon double bonds. Also, aliphatic carbon atoms were
shown in the range of 28.9-66.1 ppm.
1
1
13
Similarly, the structures of other pyrrole derivatives were determined by IR, H NMR, C NMR,
Mass spectroscopy and elemental analysis.
Scheme 3 Possible tautomeric forms of compounds 1e and 1i.
3
.2. Cytotoxicity activities
All of these new synthesized pyrrole derivatives were screened for biological properties as anticancer
activity against three human cell lines including MCF7, MOLT-4 and HL-60 using MTT assay [47]. The
different concentrations of the target compounds (25, 50, 100 and 200 µM) was selected to treat with cells
and Cisplatin was used as reference drug. The cytotoxicity activity of these analogs is shown in Table 1.
The results of tested pyrroles displayed a broad range of cell growth inhibitory activity toward tested cell
1
1

ACCEPTED MANUSCRIPT
lines. Among the compounds 1a-i, compound 1a having morpholine ring at sulfonamide moiety exhibited
good inhibitory activity against tested cell lines with 39.0, 25.5 and 30.6 µM IC values for MCF7, MOLT-
50
4
and HL-60 respectively. Replacing morpholine ring (in compound 1a) with phenyl ring (compound 1b) at
sulfonamide moiety led to loss of inhibitory activity Introduction of chloro substitution to sulfonamide in
compound 1c afforded better inhibitory activity (IC = 51.2, 38.3 and 45.4 µM respectively) in comparison
50
to its counterpart 1b. In contrast, the replacement of 1,3-dimethyl barbituric acid group (compound 1c) with
barbituric acid group (compound 1d) at 4-position of pyrrole ring did not affect inhibitory activity.
Table 1 Cell growth inhibitory activity of synthetic pyrrole derivatives assessed by the MTT reduction
assay
a
IC (µM)
5
0
Compound
MCF7
MOLT-4
25.5 ± 1.1
105.5 ± 3.4
38.3 ± 5.1
42.8 ± 3.4
>200
HL-60
30.6 ± 3.6
95.2± 3.1
45.4 ± 8.5
48.1± 5.0
>200
1
1
a
39.0 ± 4.5
b
>
200
1
c
51.2 ± 6.2
61.9 ± 2.6
>200
1
d
1
e
1
f
85.3± 7.6
63.2± 2.1
40.2± 9.0
73.2± 1.3
6.3 ± 1.5
80.1± 2.7
60.3 ± 2.2
42.0± 8.1
87.0 ± 4.6
8.1 ± 1.3
1
01± 8.1
1
1
g
82.1 ± 6.2
h
65.0± 2.1
1
i
1
23.0 ± 13.6
15.1 ± 0.5
Cisplatin
a
Values represent mean ± S.E.M.
1
2

ACCEPTED MANUSCRIPT
Fig. 4. Morphological analysis of PC-12 cells in the absence and the presence of pyrrole 1a for 48 h. (A)
phase-contrast microscopic images. (B) Fluorescence microscopic images by Hoechst 33258 staining
method.
Comparison of IC50 value of 1f with those of unsubstituted analog 1e showed that insertion of chloro group
into 4-phenyl ring of sulfonamide moiety could improve cytotoxic effect. Moreover, compound 1h having
morpholine and coumarin groups at sulfonamide moiety and 4-position of pyrrole ring exhibited inhibitory
activity in target cell lines comparing with compound 1g. It indicated that the introduction of morpholine
ring to sulfonamide moiety have better anticancer activity.
After determination of the cytotoxicity of the synthesized compounds, the morphological changes in the
nuclei of PC-12 cells were examined using phase-contrast microscopy in the absence and presence of the
active compound 1a. As shown in Fig. 4A, compound 1a-treated cells lost their polyhedral shape and
became shrinking in comparison to control cells. For apoptotic detection, PC-12 cells were stained with
Hoechst 33258. As depicted in Fig. 4B, control or untreated cells appeared regular nuclei without any
fragmentation. In contrast, the active compound 1a-exposed cells exhibited chromatin condensation and
nuclear fragmentation. Morphological results revealed that the compound 1a reduced cell viability and
induced apoptosis in PC-12 cells.
3
.3. Molecular docking study
To better understanding the mechanism of anticancer activity, docking studies were performed to fit pyrrole
compounds bearing sulfonamide groups into the active site of FGFR1 (PDB code: 4ZSA) using AutoDock
4
.2. The docking results are shown in Table 2. These results exhibited that most of the tested pyrroles
displayed hydrogen bonding and hydrophobic interactions with residues present in the active site of target
protein. The binding mode of the active compound 1a with FGFR1 protein is depicted in Fig. 5. In the
binding mode, compound 1a was nicely bound to the active site of the FGFR1 protein by seven hydrogen
bonds with Ala564, Lys566, Gly485, Ser565, Arg576 and Tyr563 and hydrophobic interactions with amino
acid groups present in the active site which could be contributed to the affinity of compound 1a with
FGFR1. Molecular docking results agreed with the biological assay data, indicated that compound 1a is
possible inhibitor of FGFR1 protein.
1
3

ACCEPTED MANUSCRIPT
Table 2 Molecular docking results of the synthesized compounds with FGFR1
H-bonds
∆
G
Compound
Distance (Å)
(
kcal/mol)
Amino acids
1
a
-8.91
Ala564, Lys566, Gly485, Ser565,
Arg576 and Tyr563
3.1, 2.8, 2.8, 2.7, 3.0,
3.3 (and 3.5)
1
b
-7.82
Glu571, Asn568, Ser565, Tyr563,
Lys482
3.5, 2.7, 2.8, 2.9, 3.5
1
c
-8.63
-8.12
Lys482, Gly485, Glu486, Asn568
3.0, 2.7, 2.8, 3.0
1
d
Asn568, Gly485, Glu486, Pro483,
Lys482
3.0, 3.0, 2.8, 3.8, 3.0
1
e
-7.35
-7.95
-8.51
-8.24
-8.08
Ser565, Tyr563, Glu486,
Asn568
3.0, 2.8, 3.3
2.7
1
f
1
g
Gly567
2.9
1
h
Glu571, Gly567
Asn568, Glu571
3.3, 2.9
2.6, 3.5
1
i
Fig. 5. Presentation of the binding mode for 1a as the most active compound in the active site of FGFR1
PDB code: 4ZSA).
(
3
.4. Computations
All calculations were performed with the Gaussian 09 program package [48]. The geometry of compound
a was fully optimized without imposing any symmetry constraint with the Becke’s three-parameter hybrid
1
functional with the Lee– Yang–Parr correlation functional (B3LYP) [49–51]. The 6-31G (d,p) [52] was
employed for the atoms of 1a [53]. The optimized structure of 1a was showed in labeled Fig. 6. Frequency
was calculated at DFT level, scaled and compared with the experimental frequency to check whether
stationary points from the geometry optimization calculations were in real minima. As shown in Table 3,
1
4

ACCEPTED MANUSCRIPT
the calculated bond lengths and bond angles at the level of theory are in reasonable agreement with the
crystallographic data.
Fig. 6. Optimized structure of compound 1a
Table 3 Some selected bond lengths (Å) and bond angles (º) of compound 1a and the experimental data
a
Bond length (Å)
C=C (aromatic)
Calculated
1.39
Experimental
1.37–1.38
N –C1
5
1.41
1.41–1.46
C=O (ester)
C=O (amide)
S –N
1.23
1.257-1.287
1.257-1.287
1.613-1.657
1.428-1.439
1.24
1.88
4
5
29
S =O , S =O
47
1.64
4
5
46
45
Bond angles (º)
O –S –O
118.82
102.61
105.39
125.61
121.5
118.86
107.77
46
45
47
29
22
C –S –N
2
2
45
O –S –C
108.05
46
45
C =C –C
6
121.69–121.67
120.8
1
2
C –C –S
2
1
22
45
C –C –C
23
120.32
124.85
120.25
1
9
18
O –C –C
3
125.6
36
35
1
5

ACCEPTED MANUSCRIPT
C –O –C
38
116.20
117.5
3
5
37
a
Ref. [54-61].
1
13
1
13
H and C NMR in DMSO were calculated with GIAO [62, 63]. IR, H NMR and C NMR chemical
shifts calculated at the B3LYP /6-31+G (d,p) level basis set and compared with experimental data.
An important parameter to measure reactivity of the molecules is the energy gap, ꢁE (ꢁE = ELUMO–EHOMO).
Decreasing in ꢁE of the molecule leads to decrease the required energy to remove an electron from the last
occupied orbital. A molecule with a low energy gap is usually more polarisable with high chemical activity,
low kinetic stability and high softness value [64]. Fig. 7 shows the 3D plots of HOMO and LUMO orbitals
of the compound 1a and gap energy.
1
a
LUMO
ꢁE(eV)
3.198
HOMO
Fig. 7. 3D plots of the HOMO, LUMO orbitals of compound 1a and ꢁE (The enery gap between HOMO and LUMO)
eV)
(
In addition, DFT is very useful in providing chemical descriptors such as chemical hardness (η),
electronegativity (χ), softness (S) and electrophilicity index (ω) Zhou & Navangul (1990) reported the
1
6

ACCEPTED MANUSCRIPT
principle of maximum hardness (absolute hardness) η, for an N-electron system with total energy E and η
are defined as:
(
ELUMO-EHOMO
)
(1)
In the formula IE is the vertical ionization energy which is approximated as -EHOMO and EA for the vertical
electron affinity as -ELUMO [65]. The global softness is the inverse of chemical hardness (S= ). The electron
affinity can also be used in combination with ionization energy to give electronic chemical potential ꢂ,
negative of electron affinity (-χ) defined as the characteristic of electronegativity of molecules [66]:
χ =-
=
(ELUMO+EHOMO
)
(2)
The global electrophilicity index, ω, is calculated using the electronic chemical potential ꢂ and chemical
hardness η [67]. According to the definition this index measures the propensity of a species to accept
electrons and is defined as:
S =
(3)
Table 4 shows the gap energy, electronegativity, electrophilicity index and chemical hardness and softness
values of the compound 1a.
Table 4 HOMO–LUMO gap energy, electronegativity, electrophilicity index and chemical
hardness and softness values of compound 1a
1
a
EHOMO (ev)
ELUMO (ev)
-7.898
-4.699
3.198
ꢁ
E L-H (ev)
Electronegativity (ꢀ) (ev)
hardness (η) (ev)
6.299
1.599
-
Softness (S) (ev )
0.3126
12.405
Electrophilicity (ψ) (ev)
3
.4.1. Computational IR
1
7

ACCEPTED MANUSCRIPT
Infrared spectra were recorded on KBr Pellet and the vibrational frequencies of 1a were calculated
at the B3LYP / 6-31G+(d,p) level of theory. In order to assign the calculated frequency to the approximate
vibrational descriptor, the vibration modes have been analyzed by means of atom movements, calculated in
Cartesian coordinates. The observed and calculated vibration frequencies of the compound in the 3200–400
-1
cm region were summarized in Table 5.
-1
The observed bands at 2980.7 and 2850.4 cm are assigned to CH stretching vibrations in aliphatic
-1
moieties. The calculated wave numbers corresponding to these bands are found at 3014.4 and 2857.5 cm
-1
respectively. The CH in-plane bending vibrations can be occurred in the region 1600–1000 cm and are
mostly observed as combined with other vibrational modes. For example, the observed bands at 1165.0,
-1
1
202.0, 1451.7 and 1493.0 cm including in plane bending modes are uncovered as mixed with vibrational
modes such as νCC in ring and νCCC in ring. The CH out-of-plane bending vibrations are observed at the
-1
-1
interval 1000–650 cm [68–72]. In this study, the observed bands 1016.6-760.6 cm is assigned to the CH
-1
out-of-plane bending modes which were computed at 985.2-770.8 cm . The asymmetric and symmetric
stretching vibration of SO were appeared at 1381.7-1275.7 while were calculated at 1386.4-1276.5
2
respectively.
-1
Table 5 Experimental and scaled vibrational wavenumbers (harmonic frequency) (cm ), and assignments
of 1a utilizing (6-31G+ (d,p) basis set
1
a
Assignment
Exprimental
Scaled
3014.4
2857.5
1729.7
1678.2
1577.7
1502.4
1405.4
1386.4
1276.5
1215.6
1162.3
985.2
2
2
1
1
1
1
1
1
1
1
1
1
9
7
7
980.7
850.4
710.1
677.4
522.3
493.0
451.7
381.7
275.7
202.0
165.0
016.6
50.6
ν (C-H) Aliphatic(asymmetric)
ν (C-H) Aliphatic(symmetric)
ν (C=O)
ν (C=O)
ν (C=C)
βHCC
βHCC
SO
SO
βHCC
2
(asym)
2
(sym)
βHCC
ɣCH
ɣCH
ɣCH
963.2
60.6
770.8
40.3
764.3
2
SO (scis)
1
8

ACCEPTED MANUSCRIPT
3
.4.2. Computational NMR
The isotropic chemical shifts are frequently used as an aid in identification of organic compounds
and accurate predictions of molecular geometries are essential for reliable studies of magnetic properties.
1
3
1
The C and H NMR isotropic shielding were calculated with the GIAO method [73, 74] using the
optimized parameters obtained from B3LYP by /6-31+G (d,p) methods. The GIAO method is one of the
most common approaches for calculating nuclear magnetic shielding tensors. In order to comparison
between experimental and theoretical NMR data, this may be helpful in making correct assignments and
13
understanding the relationship between chemical shift and molecular structure. C NMR chemical shifts
calculation for further clarification of the synthesized complexes is reported. To clarify the relation between
theoretical and experimental values of NMR chemical shift constants, the experimental data are plotted
versus computed values. The impact of the solvent was taken into account using the Polarized Continuum
1
3
Model (PCM) [75]. In order to compute the C NMR chemical shifts, each couple of carbon atoms on
equivalent locations of the compound were considered as equivalent and their average of chemical shifts
1
13
were calculated. The isotropic H and C chemical shifts calculated by all DFT methods are given in Table
, 7 respectively and compared with the experimental values. As can be seen, the results obtained by using
6
all methods are in reasonable agreement with experimental values. The chemical shift changes with
methods presumably occur due to variation of the hybrid functional. The results of B3LYP method are
close to experimental data and they differ slightly from results of experiment. As can be seen, there is a
good linear relationship between experimental and theoretical B3LYP/6-31+G (d,p) chemical shifts (for
more details see SI, Fig. 30 and 31).
1
Table 6 The Experimental and calculated H NMR nuclear shielding and assignments of 1a utilizing basis
sets 6-31G+ (d,p)
1
9

ACCEPTED MANUSCRIPT
1
a
Experimental
6-31+G(d,p)
Assignment
7
7
7
7
7
7
4
3
3
3
3
2
2
3
.36
.68
.34
.27
.18
.32
.98
.90
.69
.18
.64
.78
.78
.52
.14
8.04
7.17
7.06
6.96
6.87
6.69
6.25
4.32
3.88
3.70
3.52
3.41
3.31
3.19
2.87
54
56
49
50
55
53
48
75
72,73
61
78
63
69
65
3
58
2
R = 0.971 (R = root-mean-square deviation)
1
3
Table 7 The Experimental and calculated C NMR nuclear shielding and assignments of 1a utilizing basis
sets 6-31G+ (d,p)
Experimental
6-31+G(d, p)
175.87
Assignment
C13
1
68.00
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
63.89
57.04
52.15
38.74
31.06
29.74
34.78
30.72
31.06
29.32
29.16
28.89
14.27
13.52
28.36
24.75
173.71
158.08
155.29
142.66
139.37
136.02
135.48
133.84
132.54
129.56
129.48
129.08
128.80
127.40
123.42
123.09
76.52
C35
C21
C15
C1
C19
C4
C18
C8
C7
C11
C10
C9
C23
C20
C3
C2
6
6.09
6.09
6.96
3.34
C31
C33
C44
C38
6
75.30
5
5
73.24
58.92
2
0

ACCEPTED MANUSCRIPT
4
4
2
2
8.16
6.11
8.90
8.90
58.06
56.17
35.05
34.72
C6
C30
C27
C28
2
R = 0.986 (R = root-mean-square deviation)
4
.
Conclusions
In conclusion, a new set of pyrrole derivatives bearing sulfonamide groups was synthesized and
characterized using various spectroscopic techniques. The synthesized pyrrole derivatives were evaluated
for their toxicity against MCF7, MOLT-4 and HL-60 cell lines. Among them, compound 1a possessing
morpholine ring at sulfonamide moiety displayed good inhibitory activity. The morphological analysis by
Hoechst 33258 staining test demonstrated that the active compound 1a can induce apoptosis in PC-12 cells.
Also, Molecular docking studies were performed to insert these compounds into FGFR1 active site to
predict a possible binding mode. The obtained results suggested that compound 1a may be a promising lead
for further modifications towards search of potent antineoplastic agents. In addition, DFT calculations were
used successfully to optimize the structural of compound 1a and support the IR and NMR data.
Acknowledgment:
We are thankful to Persian Gulf University Research Council for partial support of this work.
2
1

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