2-Aryl-3-(2-morpholinoethyl)thiazolidin-4-ones: Synthesis, anti-inflammatory in vivo, cytotoxicity in vitro and molecular docking studies

Article abstract of DOI:10.1016/j.ejmech.2016.04.028

Seven new 4-thiazolidinones bearing the morpholino moiety were easily synthesized by one-pot reactions of 4-(2-aminoethyl)morpholine (2-morpholinoethylamine), arenealdehydes and mercaptoacetic acid refluxing toluene for 19 h with moderate to good yields (

Full text of DOI:10.1016/j.ejmech.2016.04.028

Accepted Manuscript
2-Aryl-3-(2-morpholinoethyl)thiazolidin-4-ones: Synthesis, anti-inflammatory in vivo,
cytotoxicity in vitro and molecular docking studies
Daniela Gouvea, Flávia Aleixo Vasconcellos, Gabriele dos Anjos Berwaldt, Amilton
Seixas Neto, Gerferson Fischer, Renata Parruca Sakata, Wanda Pereira Almeida,
Wilson Cunico
PII:
S0223-5234(16)30321-X
10.1016/j.ejmech.2016.04.028
EJMECH 8543
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 9 December 2015
Revised Date: 16 March 2016
Accepted Date: 10 April 2016
Please cite this article as: D. Gouvea, F.A. Vasconcellos, G. dos Anjos Berwaldt, A.S. Neto, G. Fischer,
R.P. Sakata, W.P. Almeida, W. Cunico, 2-Aryl-3-(2-morpholinoethyl)thiazolidin-4-ones: Synthesis, anti-
inflammatory in vivo, cytotoxicity in vitro and molecular docking studies, European Journal of Medicinal
Chemistry (2016), doi: 10.1016/j.ejmech.2016.04.028.
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Graphical Abstract
2-Aryl-3-(2-morpholinoethyl)thiazolidin-4-ones: Synthesis, anti-inflammatory in
vivo, cytotoxicity in vitro and molecular docking studies.

ACCEPTED MANUSCRIPT
2-Aryl-3-(2-morpholinoethyl)thiazolidin-4-ones: Synthesis, anti-inflammatory in
vivo, cytotoxicity in vitro and molecular docking studies.
Daniela Gouveaa, Flávia Aleixo Vasconcellosa*, Gabriele dos Anjos Berwaldta, Amilton Seixas
Netob, Gerferson FischerC, Renata Parruca Sakatad, Wanda Pereira Almeidad, Wilson Cunicoa*.
a
Laboratório de Química Aplicada à Bioativos. Centro de Ciências Químicas, Farmacêuticas e de
Alimento, Universidade Federal de Pelotas,
b Laboratório do Grupo de Estudos em Doenças Transmitidas por Animais. Faculdade de Veterinária
Universidade Federal de Pelotas,
c Laboratório de Virologia. Faculdade de Veterinária., Universidade Federal de Pelotas,
d
Laboratório de Desenvolvimento de Fármacos. Instituto de Química e Faculdade de Ciências
Farmacêuticas, Universidade Estadual de Campinas.
*Corresponding authors. Tel.: +55 (53) 3275-7358; fax: +55 (53) 3275-7354.
Campus Universitário s/nº cx postal 354, Pelotas, 96010-900, RS, Brazil
For chemistry: wjcunico@yahoo.com.br
For biological tests: aleixo.fv@gmail.com

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ABSTRACT
Seven new 4-thiazolidinones bearing the morpholino moiety were easily synthesized by one-pot
reactions of 4-(2-aminoethyl)morpholine (2-morpholinoethylamine), arenealdehydes and
mercaptoacetic acid refluxing toluene for 19 hours with moderate to good yields (45-97%).
These novel compounds were fully identified and characterized by NMR spectroscopy and mass
spectrometry. Thiazolidin-4-ones in vivo anti-inflammatory activities were determined using a
croton oil-induced ear edema model of inflammation in BALB C mice. The best results were
found for compounds 4c (49.20 mmol/Kg), 4d (49.20 mmol/Kg) and 4f (52.48 mmol/Kg),
which showed the ability to decrease the ear edema in mice by 50%, 48% and 54%,
respectively, when compared to the standard drug indomethacin. In addition, the in vitro
cytotoxicity activity of thiazolidin-4-ones against Vero cells was also performed and four
compounds (4a, 4c, 4d and 4f) showed no toxic effect at 500 µg/mL. A docking simulation of
compounds into the 1Q4G (COX-1) and 4PH9 (COX-2) enzymes binding site was conducted.
This preliminary result will guide us in for further studies to improve the anti-inflammatory
activity.
Keywords: thiazolidin-4-ones, morpholine, anti-inflammatory activity, ear edema, cytotoxicity,
molecular docking.

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1. Introduction
The skin and mucous are an important protection of the immune response to external
stimuli by mechanical and chemical agents, pathogens and autoimmune response [1, 2]. The
inflammatory response is characterized at clinical level by four classic signs: heat; redness;
swelling; and pain. An exaggerated inflammatory response can cause a physiological imbalance
of the tissue and / or organ [3]. Anti-inflammatory drugs are widely used to treat inflammatory
acute and chronic conditions. The clinical use of non-steroidal anti-inflammatory (NSAIDs)
drugs is associated with significant toxicity particularly in the gastrointestinal tract and kidney
[4]. Medicinal and organic chemistry have shown a great interest to the design and production
of new bioactive compounds without these side effects [5].
In this context, the heterocyclic 4-thiazolidinone ring is an important moiety in
medicinal chemistry. Alone or when incorporated into different templates, it has been reported
to show potent biological activities [6] including in vivo anti-inflammatory activities in models
of acute inflammation such as induced paw edema and ear edema assays [7]. Hu et al 2013 [8]
also reported the significant inhibitory activity of thiazolidinones against LPS-induced TNF-α
and IL-6 expression in RAW 264.7 macrophages. Based on knowledge regarding anti-
inflammation mechanism results from the inhibition of cyclooxygenases (COX-1/COX-2) [9]
and lipoxygenase (LOX) enzymes [2, 10], thiazolidinones may provide a promising approach
for the treatment of multi-factorial disease states such as inflammation.
Compounds with morpholine scaffolds also feature broad biological activities as
antioxidants, anti-antimicrobials, antidepressants, anti-diabetics and anti-inflammatories [11].
Literature reports that the modification of the classic anti-inflammatory ibuprofen [12] and
indomethacin [13] with addition of the morpholine group increases the selective COX-2
inhibition.
Our research group has been working with the chemical and biological properties of
heterocyclic thiazolidin-4-ones for the past few years. Recently, we published thiazolidin-4-ones
with antifungal [14] and antioxidant [15] activity. In this work, we aim to report the synthesis,

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anti-inflammatory activity against ear edema and cytotoxic activity against Vero cell lines of
thiazolidin-4-ones bearing the morpholine core. Docking studies with COX-1 and COX-2 were
also reported. The compounds were designed according to a similar structure of coxibs as
shown in Figure 1.
Figure 1. Design for proposed thiazolidin-4-ones
2. Results and discussion
4-Thiazolidinone derivatives have attracted continuing interest over the years because of
their diverse biological activities, recently reviewed by Tripathi in 2014 [6a]. A literature survey
reveals that several thiazolidin-4-ones have been prepared based on different synthetic routes
and the most common procedure involves a three component reaction of an aldehyde or ketone,
a primary amine and mercaptoacetic acid, either in a one (multicomponent or one-pot) or two-
step process [6].
Initially, reaction conditions were studied for the formation of thiazolidinone 4c by a
conventional heating methodology. For this purpose, all reactions were monitored by TLC
and/or GC analysis. According to papers that we recently published, the kind of amine
(aromatic or aliphatic) influences the reaction time and the multicomponent or one-pot reaction
influences the yields: Aliphatic amines generally required less time to complete formation of
thiazolidin-4-ones than aromatic amines [14a, 16]; and the one-pot reaction is important to
avoid the formation of undesired by-product oxathiolan-5-one [17]. We reported recently that
thiazolidin-4-ones from aliphatic amines were obtained in 2 to 5 hours [14a, 15a, 18]. Therefore,
we expected a similar reaction time to the thiazinanones synthesis when the aliphatic amine 2-

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aminoethylmorpholine was used. However, to our surprise, 19 hours were needed to complete
thiazolidinone 4c formation.
So, the 4-thiazolidinone derivatives 4a-g were synthesized from the reaction of 2-
aminoethylmorpholine 2 (1 mmol) and corresponding arenealdehyde (1a-f) (1 mmol) in toluene
reflux for 3 hours to complete formation in situ of the imine. Mercaptoacetic acid (3) (2 mmol)
was then added and the mixture was continuously refluxed for more than 16 hours to
intramolecular cyclization (Scheme 1). The thiazolidin-4-ones 4a-g were obtained at moderate
to excellent yields after purification by washing with hot hexane (45-97%). The structures of
unpublished thiazolidin-4-ones 4a-g were identified and characterized by 1H and 13C NMR, with
the assistance of 2D spectra (HMBC and HSQC) and by HRMS (see experimental section).
Scheme 1. General synthesis of thiazolidin-4-ones: Reaction conditions: (i) Toluene, 110°C,
3h; (ii) HSCH2COOH, 110°C, 16h.
To investigate the anti-inflammatory activity, the phorbol ester 12-o-
tetradecanoylphorbol-13-acetato (TPA) was used. It was present in croton oil (Croton tiglium
L.), a potent phlogistic agent and tumor promoter. Croton oil promotes an intense inflammatory
and hyperproliferative response, resembling some skin diseases [19, 20]. Topical application of
croton oil/TPA promotes an acute inflammatory reaction characterized by vasodilation,
polymorphonuclear leukocyte infiltration into the tissue and edema formation [3a, 21]. Ear
thickness was measured 6 hours after the administration of the compounds. Indomethacin was
used as a standard drug [3a].

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Table 1 shows the result for the anti-inflammatory activity of thiazolidin-4-ones 4a-g.
All compounds reduce the ear inflammation at 166 mg/Kg (39-58%). The best activity was
found for compounds 4c and 4d, both with electron-withdrawing substituent nitro, and for
compound 4f, with electron-donating substituent methoxy, which showed to be able to decrease
the ear edema in mice, by 50%, 48% and 54%, respectively. For this reason, it was difficult to
determine a structure-activity relationship for the aryl group substituent attached to
thiazolidinones. This result suggests aryl has little electronic influence of the activity. These
results were similar to those found for the standard drug Indomethacin that reduced the ear
inflammation by 58%.
It should be noted that the thiazolidin-4-ones 4a-g at doses up to the highest
concentration tested (333 mg/Kg), did not show the classical symptoms of inflammation, such
as heat, redness and swelling when compared to induced control with a phlogistic agent (acute
inflammation). Nowadays, it is difficult to treat inflammatory skin diseases because they are
chronic and require a lengthy treatment which could result in the development of drug
resistance. The most common treatments used are based on corticosteroids and recently,
treatments such as monoclonal antibodies against immunoglobulin E have also been used [21].
In addition (Table 1), the compounds showed low in vitro toxic effects on the Vero cell
line, especially thiazolidin-4-ones 4a, 4d and 4f that maintain the cell viability of 89, 92 and
89% at the highest concentration tested (1000 µg/mL). Compounds 4b, 4e and 4g showed
toxicity at 500 µ/mL, reducing cell viability by more than 20%.
Table 1. Anti-inflammatory (in vivo) and cytotoxicity activities (in vitro) of thiazolidin-4-ones
4a-g.
Cytotoxicity
Ear edema
Cell viability (µg/mL) (%)
Dose b
(mmol/Kg) activityc
Anti-infl.
Comp.
R
M.W.a
62.5
125 250 500 1000
4-F
4-Cl
2-NO2
310.39
326.84
337.39
53.48
50.79
49.20
39
44
50
100
87
98
100 100
91
76
91
89
74
83
4a
4b
4c
87
98
87
98

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3-NO2
4-OH
337.39
308.04
49.20
53.88
51.48
54.17
9.22
48
45
54
42
58
100
83
100 100
83 83
100 100
87 80
92
79
90
76
92
73
89
73
4d
4e
4-OCH3 322.42
100
87
4f
4g
4-CH3
306.42
Indomethacin
357.77
a g/moL molecular weight; b 166 mg/Kg; c % at 6h of anti-inflammatory activity.
The importance of these anti-inflammatory activities led us to perform a study of
molecular docking in cyclooxigenase-1 (COX-1) and cyclooxigenase-2 (COX-2) enzymes to
verify a comparison with a standard celecoxib drug. Results are shown in Table 2 for COX-1
and Table 3 for COX-2.
Table 2. Interactions of ibuprofen, indometacin, celecoxib and thiazolidin-4-ones 4a-g with
COX-1.
Dock score
(Kcal/mol)
-8.1
N of
interactions
3
Compound
Ibuprofen
Distance (Â)
aa
2.9
3.1
Arg120
Arg120
COOH
COOH
2.8
Tyr355
COOH
Indometacin
Celecoxib
-7.8
-6.6
3
3
4.0
3.7
3.1
3.7
2.7
3.1
3.2
3.8
Arg120
Tyr355
Tyr355
His513
Tyr355
Tyr355
Arg120
Arg120
N
OCH3
COOH
NH2
SO2
SO2
-6.5
-6.4
2
2
O morph
N morph
4a
4b
3.2
Arg120
N morph
3.8
3.1
3.8
Arg120
Arg120
Tyr355
N morph
NO2
NO2
-6.6
-6.6
2
2
4c
4d
3.1
Arg120
N morph
3.8
-
Arg120
-
N morph
-
-6.0
-6.3
0
2
4e
4f
3.1
Arg120
O morph.
3.8
3.3
Arg120
Arg120
N morph.
N morph.
-6.7
1
4g
Table 3. Interactions of ibuprofen, indometacin, celecoxib and thiazolidin-4-ones 4a-g with
COX-2.
Dock score
(Kcal/mol)
-7.6
N of
interactions
2
Compound
Ibuprofen
Distance (Â)
2.4
aa
Tyr355
-C(O)OH

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3.0
3.1
3.7
3.7
3.4
2.1
2.7
2.2
3.4
Arg120
Ser353
His90
-C(O)OH
OCH3
OCH3
NH2
Indometacin
Celecoxib
-7.7
2
6
-10.6
His90
Arg513
Gln192
Leu352
Ser353
Tyr355
SO
NH2
NH2
NH2
NN
-6.3
-6.4
2
2
4a
4b
3.0
3.3
3.0
3.3
Arg120
Arg120
Arg120
Arg120
O morph.
O thiaz.
O morph.
O thiaz.
-6.7
-7.6
1
6
3.6
2.9
3.9
3.4
3.7
3.7
3.6
3.7
3.3
2.8
3.2
Arg120
Tyr355
Arg120
His90
Ser353
Met523
Ser530
His90
NO2
NO2
NO2
4c
4d
O morph.
O and N morph.
O thiaz.
S thiaz.
OH
-6.8
-6.9
3
3
4e
4f
Arg120
O morph.
His90
Tyr355
OH
O morph.
3.3
3.8
3.0
3.3
Ser353
Ser530
Arg120
Arg120
O and N morph.
S of thiaz.
O morph.
-6.6
2
4g
O thiaz.
Celecoxib shows six interactions with COX-2 (Figure 2): a) four hydrogen bonds are
formed between NH2 with amino acids His90, Gln192, Leu352 and Ser353 at distances of 3.7,
2.1, 2.7 and 2.2 Å, respectively; b) hydrogen bonds between SO with Arg513 at 3.4 Å and; c) N
of pyrazole ring with Tyr355 at 3.4 Å. In COX-1, the celecoxib shows only two hydrogen bonds
between NH2 with His513 (3.7Å) and one hydrogen bond of SO2 with Tyr355 (2.7Å) (see
supplementary information).

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Fig. 2. Molecular docking of celecoxib binding site of COX-2 (dock score -10.6
kcal/mol) containing six hydrogen bonds as yellow dotted lines.
According to the docking analyses of thiazolidin-4-ones 4a-g, the nature and position of
the substituent in the phenyl ring induce different interactions with COX-2 (Figure 3).
Fig. 3. Molecular docking of thiazolidin-4-ones 4a-g binding COX-2. 4a (White); 4b (orange);
4c (blue); 4d (yellow); 4e (red); 4f (pink); 4g (purple).
Thiazolidin-4-ones 4d, 4e and 4f exhibit the greatest number of interactions with COX-
2 amino acids. Compound 4d, with nitro group at m-position, shows six hydrogen bonds

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(Figure 4): a) the O of morpholine ring with His90 (3.4 Å), as NH2 in celecoxib; b) the O and N
of the morpholine ring with Ser353 (3.2 and 3.7 Å), as NH2 in celecoxib; c) the NO2 group with
Tyr355 (2.9 Å), as NN of pyrazole in celecoxib; d) the NO2 with Arg120 (3.9 Å) and; e) the S
of the thiazolidinone moiety with Ser530 (3.5 Å). This compound (4d) exhibits only two
interactions with COX-1 (see supplementary information) through two hydrogen bonds between
the N of morpholine and Arg120 with a distance of 3.1 and 3.8 Å.
Fig. 4. Molecular docking of 4d binding site of COX-2 with dock score -7.6 kcal/mol
exhibits six hydrogen bonds illustrated as yellow dotted lines.
Changing the NO2 group from the meta position (4d) to the orto position (4c) in the
phenyl ring, changes the binding mode of thiazolidin-4-one 4c. Compound 4c shows only one
hydrogen bond between the NO2 group and Arg120 at 3.6 Å (Table 3). The COX-1 docking of
compound 4c shows two hydrogen bonds between the NO2 and Arg120 and Tyr355 at 3.1 and
3.8 Å, respectively (Table 2).
The thiazolidin-4-one 4f with substituent 4-OCH3, exhibits COX-2 dock score -6.9
Kcal/mol, containing hydrogen bonds between O and N of morpholine with Ser353, O of
thiazolidinone with Met523 (3.7 Å) and S of thiazolidinone with Ser530 (3.8 Å) (Table 3). This
compound also shows one hydrogen bond between the N of morpholine and Arg120 (3.3 and
3.8 Å) (Table 2).

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The thiazolidinone 4e (R=4-OH) shows the COX-2 docking score -6.8 kcal/mol and
reveals hydrophilic interactions (H-bonds) between the hydroxyl group and Tyr355 and His90
at 2.8 and 3.7 Å. The hydrogen bond between the O of morpholine and Arg120 at 3.3 Å of
distance is also observed (Table 3). This compound does not show interaction (less than 4 Å)
with amino acids of the COX-1 enzyme.
The compounds 4a, 4b and 4g, with substituents 4-F, 4-Cl and 4-CH3, show dock scores
-6.3, -6.4 and -6.6 Kcal/mol, respectively. According to docking, these compounds overlapped
(Figure 3) and show the same type of interaction with COX-2 enzyme: hydrophilic interaction
between Arg120 and O of the morpholine ring and O of thiazolidinone at 3.0 and 3.3 Å of
distance (Table 3). These compounds also show interaction with the COX-1 enzyme through
hydrogen bonds between N of morpholine and Arg120 (Table 2).
These in silico results suggest that some thiazolidin-4-ones may be COX-2 selective,
especially thiazolidinone 4d, however, enzyme studies were necessary to confirm this
hypothesis.
3. Conclusion
In summary, seven novel thiazolidin-4-ones were easily synthesized by one-pot
multicomponent reaction in good yields. In addition, two thiazolidin-4-ones (4d and 4f) showed
in vivo anti-inflammatory activity, reducing the ear edema inflammation up to 50% with no
cytotoxic effect at 1000 µg/mL. The molecular docking showed that compound 4d has six
hydrogen bonds with six different amino acids, three of them like celecoxib had. These
important findings will guide us to further chemical modifications for the development of new
compounds to be employed as biologically useful agents.
4. Experimental
4.1. Chemistry

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The reactions were monitored by thin layer chromatography (TLC, using silica gel
60F253 aluminum sheets; visualization by ultraviolet light 254 nm; (hexane / ethyl acetate 3:1)
and/or by a Shimadzu Gas Chromatograph GC-2010, Column L.D., 0,25 nm; Column length, 30
m. Column Head Pressure, 14 psi, program: T°= 50 °C; t° = 2.0 min; rate 16.0 °C/min; Tf = 250
°C; tf = 10.0 min; Injector = 250 °C; Detector = 270 °C. Melting points were determined using
open capillaries on a Fisatom model 430 apparatus and are uncorrected. GC–MS analyses were
performed on a GC 2010-plus GC–MS-QP2010SE System AOC-20i – auto injector. 1H and 13C
NMR spectra were recorded using a Bruker DRX 400 spectrometer (1H at 400.14 and 500 MHz
and 13C at 100.61 MHz) or a Bruker Avance 500 spectrometer (1H at 500.13 MHz and 13C at
13
125.75 MHz) or a Bruker Ac-200F spectrometer (1H at 200.13 MHz and C at 50.32 MHz) in
CDCl3 containing TMS as in internal standard. For compound 4a, the 13C NMR was recorded at
a Bruker DRX 400 spectrometer (13C at 100.61 MHz). Infrared spectra (IR) were obtained in
ATR on an IR Agilent Absorption Spectrometer, model CARY 630 FTIR. The elemental
analyses (C, H, N) were used for all compounds on a CHNS 2400 Perkin Elmer Analyses
equipment. Analyses were consistent within ± 0.4 theoretical values.
4.1.1. General synthesis of 2-aryl-3-(2-morpholinoethyl)thiazolidin-4-ones 4a-g
To a flask containing toluene (70 mL) attached to a Dean–Stark trap, 2-
morpholinoethylamine 1 (1 mmol) and corresponding arenealdehyde 2a-f (1 mmol) were added.
The mixture was refluxed for 3 hours, the mercaptoacetic acid 3 (2 mmol) was added and the
mixture was refluxed for 13 hours more. The organic layer was washed with a saturated solution
of NaHCO3 (3 x 30 mL), dried with MgSO4 and concentrated to give the products. When
necessary, the crude products were washed with hot hexane to furnish the pure thiazolidin-4-
ones.
4.1.1.1. Selected data for 2-(4-fluorophenyl)-3-(2-morpholinoethyl)thiazolidin-4-one 4a
1
Yield 60% of the yellow solid, m.p. 90-93 °C. H NMR δ (200 MHz, ppm, JH-H = Hz); 7.30 (d,
3
3
2
2H, J=8.6); 7.08 (t, 2H, J=8.6); 5.85 (s, 1H, H2); 3.80 (dt, 1H, J=12.4,3J=6.1); 3.79 (dd, 1H,

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4
2
3
2
2J=14.4, J=1.7, H5a); 3.70 (d, 1H, J=14.7, H5b); 3.67 (t, 4H, J=4.6); 2.78 (dt, 1H, J=12.6,
1
3J=6.3); 2.55-2.31 (m, 6H). 13C NMR δ (50 MHz, ppm); 171.2 (C4); 137.8 (d, JC-F=242.6);
128.9, (d, 3JC-F=8.2); 11.,4 (d, 2JC-F=21.9); 66.8; 63.3 (C2); 55.8; 53.8; 39.3; 32.9 (C5). IR (ATR)
V max/cm-1: 2950 (C-H sp2); 2819 (C-H sp3); 1653 (C=O); 1115 (C-O). GC-MS m/z (%): 310
(M+, 2); 281 (5); 207 (12); 153 (2.5); 139 (3); 113 (7); 100 (100); 56 (12.5); 44 (32.5). HRMS-
[ESI] for C15H19FN2O2S (M+ H)+: 307.1475; found 307.1475. Anal. Calc. for C15H19FN2O2S; C,
58.04; H, 6.17; N, 9.03. Found. C. 57.49; H, 5.98; N, 8.86.
4.1.1.2. Selected data for 2-(4-clorophenyl)-3-(2-morpholinoethyl)thiazolidin-4-one 4b
Yield 76% of the white solid, m.p. 50-53 °C, 1H NMR δ (200 MHz, ppm, JH-H = Hz); 7.37 (d,
2H, 3J=8.5); 7.24 (t, 2H, 3J=8.5); 5.84 (s, 1H, H2); 3.82 (dt, 1H, 2J=14.3, 3J=6.1); 3.75 (dd, 1H,
4
2
3
2
2J=15.0, J=1.7, H5a); 3.73 (dt, 1H, J=14.4,3J=6.4); 3.71 (t, 4H, J=4.6); 3.70 (d, J=14.4,
2
3
3
2
3
H5b); 2.52 (dt, 1H, J=12.8, J=6.4); 2.42 (t, 4H, J=4.3); 2.32 (dt, 1H, J=12.9, J=5.9). 13C
NMR δ (50 MHz, ppm); 171.2 (C4); 138.2; 134.9, 129.3; 128.3; 66.6 (2C); 60.2 (C2); 56.2;
53.6 (2C); 39.6; 32.8 (C5). IR (ATR) V max/cm-1: 2948 (C-H sp2); 2855 (C-H sp3); 1670
(C=O); 1171 (C-O). GC-MS m/z (%): 326 (M+, 2); 207 (5); 155 (5); 113 (3); 98 (100); 96 (4);
69 (5). C15H19ClN2O2S (M+ H)+: 327.0929; found 327.0931. Anal. Calc. for, C15H19ClN2O2S: C,
55.12; H, 5.86; N, 8.57. Found: C, 55.09; H, 5.87; N, 8.38.
4.1.1.3. Selected data for 3-(2-morpholinoethyl)-2-(2-nitrophenyl)thiazolidin-4-one 4c
Yield 45% of the white solid, m.p. 104-106 °C, 1H NMR δ (200 MHz, ppm, JH-H = Hz); 8.11 (d,
3
3
3
3
2H, J=8.1); 7.72 (t, 2H, J=7,7); 7.52 (t, 1H, J=8.3, 4J=1.2); 7.32 (t, 1H, J=7.9), 6.60 (s, 1H,
2
2
4
2
H2); 3.97 (dt, 1H, J=14.3, 3J=5.3); 3.74 (dd, 1H, J=15.4, J=1.8, H5a); 3.61 (d, 1H, J=15.6);
3.76 (dt, 1H, 2J=13.7, 3J=6.9); 3.58 (q, 4H, 3J=3.9); 2.81 (dt, 1H, 2J=14.3, 3J=5.0); 2.57 (dt, 1H,
3
2
3
3
2J=12.4, J=5.1); 2.44 (dt, 1H, J=12.4, J=5.1); 2.44 (t, 4H, J=4.3).13C NMR δ (50 MHz,
ppm); 172.3 (C4); 136.7; 134.3, 129.1, 126.1; 115.9; 66.9 (2C); 63.2 (C2); 58.8; 53.6 (2C);
39,4; 32,7 (C5). IR (ATR) V max/cm-1: 2970 (C-H sp2); 2929 (C-H sp3); 1653 (C=O); 1521 and
1346 (NO2); 1115 (C-O). GC-MS m/z (%): 281 (M+-56, 2); 220 (5); 207 (14); 113 (5); 100
(100); 70 (5); 56 (10). HRMS- [ESI] for C15H19N3O4S (M+ H)+: 338.1169 found: 338.1175.
Anal. Calc. for C15H19N3O4S: C, 53.40; H, 5.68; N, 12.45. Found: C. 53.07; H, 5.75; N, 12.16.

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4.1.1.4. Selected data for 3-(2-morpholin-4-ethyl)-2-(3-nitrophenyl)thiazolidin-4-one 4d
Yield 93% of the white solid, m.p. 89-92 °C, 1H NMR δ (500 MHz, ppm, JH-H = Hz); 8.24 (dd,
3
4
3
3
1H, J=8.1); 8.08 (d, 1H, J=1.7); 7.58 (d, 1H, J=7.7); 7.53 (t, 1H, J=7.5); 5.90 (d, 1H,
2
3
2
3
2
4J=1.2); 3.86 (dd, 1H, J=14.5, J=5.9); 3.80 (dt, 1H, J=14.5, J=5.9); 3.86 (dd, 1H, J=15.3,
4J=1.6); 3.77 (d, 1H, 2J=15.5); 3.70 (t, 4H, 3J=4.6); 2.69 (dt, 1H, 2J=14.5, 3J=6.6); 2.44 (dt, 2H,
3
3
13
2J=13.0, J=6.3); 2.32 (q, 4H, J=5.7). C NMR δ (125 MHz, ppm); 171.3 (C4); 148.7; 132.8;
130.4; 124.1; 122.2; 66.9 (2C); 62.9 (C2); 56.0; 53.7 (2C); 39.6; 32.8 (C5). IR (ATR) V
max/cm-1: 3050 (C-H sp2); 2950 (C-H sp3); 1650 (C=O); 1500 and 1300 (NO2); 1100 (C-O).
GC-MS m/z (%): 281 (M+-56, 4%); 253 (3); 207 (14); 113 (3); 100 (100); 70 (8); 56 (14).
HRMS- [ESI] for C15H19N3O4S (M+ H)+: 338.1169; found 338.1171. Anal. Calc. for
C15H19N3O4S: C, 53.40; H, 5.68; N, 12.45. Found. C. 52.94; H, 5.46; N, 11.96.
4.1.1.5. Selected data for 2-(4-hidroxyphenyl)-3-(2-morpholinoethyl)thiazolidin-4-one 4e
1
Yield 65% of the white solid, m.p. 138-141 °C, H NMR δ (400 MHz, ppm, JH-H = Hz); 7.09
3
4
2
(dd, 3H, 3J=6.1, 4J=1.5); 6.72 (dd, 1H, J=8.5, J=1.6); 5.69 (s, 1H, H2); 3.76 (dt, 1H, J=14.4,
2
2
3
3J=6.0); 3.74 (dd, 1H, J=15.7,4J=1.8, H5a); 3.67 (d, 1H, J=15.4, H5b); 3.61 (t, 4H, J=4.9);
2
3
2
3
2.78 (dt, 1H, J=14.4, J=6.3); 2.42 (dt, 2H, J=12.9, J=6.6); 2.45-2.24 (m, 5H).13C NMR δ
(100 MHz, ppm); 171,4 (C4); 141.9; 135.1; 130.4; 129.3; 127.1; 125,1; 66.9 (2C); 63.3 (C2);
55.9; 53.6 (2C); 39.5; 32.8 (C5). IR (ATR) V max/cm-1: 3186 (OH); 2959 (C-H sp2); 2924 (C-H
sp3); 1633 (C=O); 1113 (C-O). GC-MS m/z (%): 308 (M+, 2); 280 (2); 207 (3); 113 (6); 100
(100); 56 (7). HRMS- [ESI] for C15H20N2O3S (M+ H)+: 309.1267 found: 309.1272.
4.1.1.6. Selected data for 2-(4-methoxylphenyl)-3-(2-morpholinoethyl)thiazolidin-4-one 4f
1
Yield 96% of the yellow solid, m.p. 40-43 °C. H NMR δ (500 MHz, ppm, JH-H = Hz); 7.25 (d,
3
3
2
2H, J=8.3); 6.90 (d, 2H, J=8.7); 5.80 (s, 1H, H2); 3.87 (s, 1H, OCH3); 2.81 (dt, 1H, J=13.1,
2
2
3
3J=8.7); 3.76 (dd, 1H, J=15.9,4J=1.6, H5a); 3.75 (d, 1H, J=16.8, H5b); 3.68 (t, 4H, J=4.6);
2
3
2
3
2
3
2.81 (dt, 1H, J=12.7, J=7.1); 2.50 (dt, 1H, J=12.8, J=6.4); 2.37 (dt, 2H, J=12.8, J=6.4);
3
2.40 (t, 4H, J=5.7). 13C NMR δ (125 MHz, ppm); 171.2 (C4); 160.2; 138.2; 134.9; 129.6;
128.3; 66.9 (2C); 63.8 (C2); 55.8; 53.8 (2C); 39.5; 32.8 (C5). IR (ATR) V max/cm-1: 2949 (C-H
sp2); 2850 (C-H sp3); 1665 (C=O); 1115 (C-O). GC-MS m/z (%): 322 (M+, 5); 208 (2); 151 (3);

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113 (17); 100 (100); 86 (10); 70 (7); 56 (12). HRMS- [ESI] for C16H22N2O2S (M+ H)+:
307.1475 found: 307.1475.
4.1.1.7. Selected data for 2-(4-methylphenyl)-3-(2-morpholinoethyl)thiazolidin-4-one 4g
Yield 89% of the white solid, m.p. 105-107 °C. 1H NMR δ (400 MHz, ppm, JH-H = Hz); 7.21 (s,
4
2
3
2
4H); 5.86 (d, 1H, J=1.2, H2); 3,83 (dt, 1H, J=14.1, J=6.2); 3.82 (dd, 1H, J=15.6,4J=2.0);
3.73 (d, 1H, 2J=15.5); 3.67 (t, 4H, 3J=4.6); 2.83 (dt, 1H, 2J=14.3, 3J=6.7); 2.51 (dt, 2H, 2J=12.8,
3J=6.4); 2.36 (m, 9H). 13C NMR δ (100 MHz, ppm); 171.4 (C4); 139.2; 136.4; 129.8; 126.9;
67.0 (C2); 63.9; 55.9; 53.6; 39.4; 33.0 (C5); 21.4 (Me). IR (ATR) V max/cm-1: 3166 (C-H sp2);
1633 (C=O). GC-MS m/z (%): 306 (M+, 5); 192 (2); 135 (5); 113 (15); 100 (100); 86 (12); 70
(6); 56 (12). HRMS- [ESI] for C16H22N2O2S (M+ H)+: 307.1475 found: 307.1475. Anal. Calc.
For C16H22N2O2S, C, 62.71; H, 7.23; N, 9.09. Found. C. 62.69; H, 7.43; N, 9.14.
4.2. Animals
Experiments were performed on BALB/c male mice (25-35 g). The animals were
controlled at room temperature (22 ± 3 °C) under 8 h, with access to water and food ad libitum.
The experimental protocols in this study were approved and followed in accordance with the
guidelines of the Ethical Committee in Animal Experimentation of the Federal University of
Pelotas (CEEA 7068), in accordance with international guidelines.
4.3. Determination of anti-inflammatory effect in ear edema
BALB/c mice weighing 30 g, in mean, of were divided into three animals in each group.
Edema was induced of croton oil at a concentration of 2 µg/ear, dissolved in acetone. The
compounds 4a, 4b, 4c, 4d, 4e, 4f and 4g at 33, 100, 166 and 333 mg/Kg concentrations and
indomethacin at 33 mg/Kg concentration (a positive control). All the suspensions were prepared
in the vehicle acetone (20%) and administered by topical application in a constant volume of 20
µl per ear mouse. The anti-inflammatory potency of thiazolidin-4-ones was compared with the
used standard drug, Indomethacin [22].

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The edema was expressed as an increase in ear thickness due to inflammation. Ear
thickness was measured before and after inflammatory induction. A 6 mm section of ears was
obtained and weighed analytical balance (BEL engineering Mark M214A). The swelling
induced was assessed as the increase in weight of ear punch of treated groups over untreated one
and it was called the edema index, according to the method reported by Tubaro et al 1985 [19].
Croton oil-induced ear inflammation, carry out all experiments between 9 a.m. and 5
p.m., in order to avoid the influence of circadian variations in corticosteroid levels in the
inflammatory responses [23].
4.4. Inhibition of inflammation analysis
The percentage protection of edema (inhibition of inflammation) was calculated
according to the formula percentage anti-inflammatory activity = 100 x (1 - Vt/Vc), where Vt and
Vc are the volume of edema in test compounds and control groups edema, respectively [24].
Measure was obtained at 6 h after application of compounds [5b, 7b].
4.5. Cytotoxicity assay (MTT assay)
The Vero cells (African green monkey kidney cell line) were used to analyze toxicity of
compounds. The culture medium used to culture and compounds diluted was the modified
Eagle´s medium (E-MEM, Sigma-Aldrich, USA) supplemented with penicillin (Sigma Aldrich,
USA), streptomycin (Vetec, Brazil), amphotericin B (Cristália, Brazil) and enrofloxacin (Bayer,
Brazil). The Vero cells were cultured in 96-well plates (2 x 106 cells mL-1) in E-MEM
supplemented with 10% (v/v) fetal bovine serum (Gibco, USA) and were allowed to attach for
24 h and 48 h before treatment with compounds. Series dilutions were prepared, ranging
between 62.5, 125, 250, 500 and 1000 µg/mL of a previous solution in E-MEM. After removing
the test product, cells were washed with phosphate-buffered saline (PBS). Then, as described by
Mosmann (1983) [25] and Scopel e Silva [26], 50 mL of MTT (3-(4,5-dimethylthiazol-2-yl)-

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2,5-diphenyltetrazolium bromide) solution 1 mg/mL in PBS, was added to each to each well.
After 4h of incubation at 37 °C in darkness, the supernatant was replaced to 100 mL of
dimethylsulfoxide (DMSO, Synth®, purity 99.9%) to solubilize the formazan salt generated by
the reaction between salt MTT and mitochondrial metabolism of viable cell. The plates were
gently shaken for 10 minutes, and the colorimetric intensity was evaluated by
spectrophotometry wavelength of 540 nm. All experiments were performed three times. The
percentage of viability was calculated as AT/AC x 100; where AT and AC are the absorbance of
treated and control cells, respectively.
4.6. Molecular docking
The molecular docking studies of thiazolidin-4-ones 4a-g were performed in AutoDock
Vina program [27]. Firstly, the compounds were sketched in GaussView 5.0.8 program, then
compounds ideal conformations were calculated by semi-empirical method PM6 in Gaussian 09
program and the obtained structures were used to perform the docking. The crystal structures of
the enzymes Cyclooxygenase-1 (COX-1) and Cyclooxygenase-2 (COX-2) were obtained from
PDB (Protein Data Bank) with codes 1Q4G and 4PH9 respectively. The 1Q4G enzyme is
complexed with 2-(1,1'-biphenyl-4-yl) propanoic acid. This enzyme is from Ovis aries organism
and the resolution of the crystal Structure is 2.00 Å. The 4PH9 enzyme is complexed with
ibuprofen, this enzyme is from Mus musculus organism and the resolution of the crystal
Structure is 1.81 Å. As enzyme preparation, the water molecules were removed and added polar
hydrogens only. In the AutoDock Vina, the grid size to perform docking in 1Q4G was x = 26; y
= 26; z = 26 and the coordinates was x = 27.772; y = 34.177; z = 201.688. To perform docking
in 4PH9 the grid size was x = 26; y = 26; z = 26 and the coordinates was x = 11.677; y =
23.272; z = 25.420. To validate the methodology, it was done redoking. Poses obtained by
redocking of 2-(1,1'-biphenyl-4-yl)propanoic acid and ibuprofen are close to crystal binders

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indicating that the docking conditions are correct. The results were analyzed using the PyMOL
Molecular Graphics System, Version 1.7.2.1.
Supplementary information
1
The supplementary information contains the H and 13C NMR, Infrared and CG-MS
spectrum for thiazolidin-4-ones 4a-g.
Acknowledgements
The authors thank UFPel, FAPERGS (11/2068-7) and CAPES (23038.007284/2012-
02) for financial support. UNICAMP and CNPq are also acknowledged. The authors also thank
Prof. Hedy Hofmann for the help with the manuscript
References
[1] T. Welss, D. A. Basketter, K. R. Schröder, In vitro skin irritation: facts and future. State of
the art review of mechanisms and models, Toxicology in Vitro 18 (2004) 231–243.
[2] A. A. Geronikaki, A. A. Lagunin, D. I. Hadjipavlou-Litina, P. T. Eleftheriou, D. A.
Filimonov, V. V. Poroikov, I. Alam, A. K. Saxena, Computer-aided discovery of anti-
inflammatory thiazolidinones with dual cyclooxygenase/lipoxygenase inhibition, J. Med. Chem.
51 (2008) 1601–1609.
[3] a) E. J. Joung, M. S. Lee, J. W. Choi, J. S. Kim, T. Shin, B. M. Jeong, N. Y. Yoon, C. W.
Lim, J. Kim, H. R. Kim, Anti-inflammatory effect of ethanolic extract from Myagropsis
myagroides on murine macrophages and mouse ear edema, BMC.Complementary Altern. Med.
12 (2012) 171-181. b) C. N. Serhan, J. Savill, Resolution of inflammation: the beginning
programs the end, Nature Immunol. 6 (2005) 1191-1197.
[4] F. T. Uchôa, T. G. Silva, M. C. A. Lima, S. L. Galdino, I. R. Pitta, T. D. Costa, Preclinical
pharmacokinetic and pharmacodynamic evaluation of thiazolidinone PG15: an anti-
inflammatory candidate, J. Pharm. Pharmacol. 61 (2009) 339–345.
[5] a) V. Kumar, A. P. C. Sharma, Synthesis of some novel 2,5-disubstituted thiazolidinones
from a long chain fatty acid as possible anti-inflammatory, analgesic and hydrogen peroxide
scavenging agents, J. Enzyme Inhib. Med. Chem. 26 (2011) 198–203. b) A. Sharma, V. Kumar,
S. Jain, and P. C. Sharma, Thiazolidin-4-one and hydrazone derivatives of capric acid as

ACCEPTED MANUSCRIPT
possible anti-inflammatory, analgesic and hydrogen peroxide-scavenging agents, J. Enzyme
Inhib. Med. Chem. 26 (2011) 546–552.
[6] a) A. C. Tripathi, S. J. Gupt, G. N. Fatima, P. K. Sonar, A. Verma, S. K. Saraf, 4-
Thiazolidinones: The advances continue, Eur. J. Med. Chem. 72 (2014) 52-77. b) A. K. Jain, A.
Vaidya, V. Ravichandran, S. K. Kashaw, R. K. Agrawal, Recent developments and biological
activities of thiazolidinone derivatives: A review, Bioorg. Med. Chem. 20 (2012) 3378–3395.
[7] a) I. Apostolidis, K. Liaras, A. Geronikaki, D. Hadjipavlou-Litina, A. Gavalas, M. Sokovic,
J. Glamoclija, A. Ciric, Synthesis and biological evaluation of some 5-arylidene-2-(1,3-thiazol-
2-ylimino)-1,3-thiazolidin-4-ones as dual anti-inflammatory/antimicrobial agents, Bioorg. Med.
Chem. 21 (2013) 532–539. b) C. Nandagokula, B. Poojary, S. Vittal, S. Shenoy, P. Shetty, A.
Tangavelu, Synthesis, characterization, and biological evaluation of some N-aryl hydrazones
and their 2,3-disubstituted-4-thiazolidinone derivatives, Med. Chem. Res. 22 (2013) 253–266.
c) S. H. Shelke, P. C. Mhaske, M. Nandave, S. Narkhade, N. M. Walhekar, V. D. Bobade,
Synthesis and pharmacological evaluation of a novel series of 3-aryl-2-(2-substituted-4-
methylthiazole-5-yl)thiazolidin-4-one as possible anti-inflammatory and antimicrobial agents,
Bioorg. Med. Chem. Letters 22 (2012) 6373–6376. d) V. Kumar, A. P. C. Sharma, Synthesis of
some novel 2,5-disubstituted thiazolidinones from a long chain fatty acid as possible anti-
inflammatory, analgesic and hydrogen peroxide scavenging agents, J. Enzyme Inhib. Med.
Chem. 26 (2011) 198–203. e) A. Deep, S. Jain, P. C. Sharma, P. Phogat, M. Malhotra, Synthesis
of 2-(aryl)-5-(arylidene)-4-thiazolidinone derivatives with potential analgesic and anti-
inflammatory activity, Med. Chem. Res. 21 (2012) 1652–1659.
[8] J. Hu, Y. Wang, X. Wei, X. Wu, G. Chen, G. Cao , X. Shen, X. Zhang, Q. Tang, G. Liang,
X. Li, Synthesis and biological evaluation of novel thiazolidinone derivatives as potential anti-
inflammatory agents, Eur. J. Med. Chem. 64 (2013) 292-301.
[9] a) S. G. Alegaon, M. B. Hirpara, K. R. Alagawadi, K. K. Hullatti, K. Kashniyal, Synthesis of
novel pyrazole–thiadiazole hybrid as potential and selective cyclooxygenase-2 (COX-2)
inhibitors, Bioorg. Med. Chem. Lett. 24 (2014) 5324–5329. b) A. Zarghi, E. H. Arefi, E. O. G.
Dadrass, E. S. Torabi, Design and synthesis of new 2-aryl, 3-benzyl-(1,3-oxazolidine or 1,3-
thiazolidine)-4-ones as selective cyclooxygenase (COX-2) inhibitors, Med. Chem. Res. 19
(2010) 782–793. c) E. Pontiki, D. H. Litina, K. Litinas, O. Nicolotti, A. Carotti, Design,
synthesis and pharmacobiological evaluation of novel acrylic acid derivatives acting as
lipoxygenase and cyclooxygenase-1 inhibitors with antioxidant and anti-inflammatory activities,
Eur. J. Med. Chem. 46 (2011) 191-200.
[10] a) P. Eleftheriou, A. Geronikaki, D. Hadjipavlou-Litina, P.Vicini, O. Filz, D. Filimonov, V.
Poroikov, S. S. Chaudhaery, K. K. Roy, A. K. Saxena, Fragment-based design, docking,
synthesis, biological evaluation and structure activity relationships of 2-
benzo/benzisothiazolimino-5-aryliden-4-thiazolidinones
as
cyclooxygenase/lipoxygenase

ACCEPTED MANUSCRIPT
inhibitors, Eur. J. Med. Chem. 47 (2012) 111-124. b) Bennamane, B. N. Kolli, A. A.
Geronikaki, P. Th. Eleftheriou, R. Kaoua, K. Boubekeu, P. Hoffman, S. S. Chaudhary, A. K.
Saxena, N-Substituted [phenyl-pyrazolo]-oxazin-2-thiones as COX-LOX inhibitors: influence
of the replacement of the oxo-group with thioxo-group on the COX inhibition activity of N-
substituted pyrazolo-oxazin-2-ones, ARKIVOC (2011) 69-82.
[11] V.A. Pal’chkov, Morpholine, synthesis and biological activity, Russian J. Org. Chem. 49
(2013) 787-814.
[12] J. Dou, L. Shi, A. Hu, M. Dong, J. Xu, A. Liu,Y. Jiang Synthesis and Evaluation of 2-(2-
,
Arylmorpholino)ethyl Esters of Ibuprofen Hydrochlorides as COX-2 and Serotonin Reuptake
Inhibitors, Arch. Pharm. Chem. Life Sci. 347 (2014), 89–95.
[13] L. Shi., H. Aixi, X. Jiangpina, J. Yiping. Design, synthesis and biological evaluation of 2-
(2-arylmorpholino-4-yl)ethy esters. Chin. J. Chem. 30 (2012) 1339-1344.
[14] a) A. Kunzler, P. D. Neuenfeldt, A. M. Neves, C. M. P. Pereira, G. H. Marques, P. S.
Nascente, M. H. V. Fernandes, S. O. Hubner, W. Cunico. Synthesis, antifungal and cytotoxic
activities of 2-aryl-3-((piperidin-1-yl)ethyl) thiazolidinones. Eur. J. Med. Chem. 64 (2013) 74-
80. b) G. H. Marques, G. G. Nascimento, A. Kunzler, V. B. O Bareno, B. B. Drawanz, H. G.
Mastelloto, F. R. M. Leite, P. Nascente, G. M. Siqueira, W Cunico, Antifungal activity of 3-
(heteroaryl-2-ylmethyl)thiazolidinone derivatives. Med. Chem. 10 (2014) 355-360.
[15] a) A. M. Neves, A. R. Duval, G. A Berwaldt, D. P. Gouvêa, N. P. Flores, P. G. Silva, F.
M.; Stefanello, W. Cunico. Thiazolidin-4-ones from 3-(aminomethyl)pyridine, arenealdehydes
and mercaptoacetic acid: Synthesis and radical scavenger activity. J. Braz. Chem. Soc. 26
(2015) 381-388. b) J. Bosenbecker, V. D. O Bareño, R. Difabio, F. A. Vasconcellos, F. S. P.
Dutra, P. S. Oliveira, A. G. Barschak, F. M. Stefanello, W. Cunico, Synthesis and antioxidant
activity of 3-(pyridin-2-ylmethyl)-1,3-thiazinan(thiazolidin)-4-ones, J. Biochem. Mol. Toxicol.
28 (2014) 425-432. c) J. C. Campos, D. P. Gouvea, C. S. Ribeiro, F. S. P Dutra, F. M.
Stefanello, C. M. P. Pereira, W. Cunico, G. M. Siqueira, Efficient synthesis and antioxidant
evaluation of 2-aryl-3-(pyrimidin-2-yl)-thiazolidinones, J. Biochem. Mol. Toxicol. 27 (2013)
445-450.
[16] D. P. Gouvêa, V. D. O. Bareño, J. Bosenbecker, B. B. Drawanz, P. D. Neuenfeldt, G. M.
Siqueira,; W. Cunico, Ultrasonics promoted synthesis of thiazolidinones from 2-aminopyridine
and 2-picolilamine, Ultrason. Sonochem. 19 (2012) 1127-1131.
[17] W. Cunico, W. T. Vellasco Junior, M. Moreth, C. R. B. Gomes, Microwave-assisted
synthesis of 1.3-thiazolidin-4-ones and 2-aryl-1,3-oxathiolan-5-ones, Lett. Org. Chem. 5 (2008)
349-352.
[18] P. D. Neuenfeldt, B. B. Drawanz, A. C. C. Aguiar, F. Figueiredo, A. Krettli, W. Cunico,
Multicomponent synthesis of new primaquine thiazolidinone derivatives, Synthesis (2011)
3866-3870.

ACCEPTED MANUSCRIPT
[19] A. Tubaro, P. Dri, G. Delbello, C. Zilli, R. D. Logia. The croton oil ear test
revisited. Agents Actions 17 (1985) 347–349.
[20] M. Gábor, Models of acute inflammation in the ear, in: P.G. Winyard, D.A. Willoughby
(Eds.), Inflammation Protocols, first ed., Szeged, Hungary, 2003, pp. 129-137.
[21] D. A. Cabrini, H. H. Moresco, P. Imazu, C. D. Silva, E. F. Pietrovski, D. A. G. B. Mendes,
A. S. Prudente, M. G. Pizzolatti, I. M. C. Brighente, M. Fleithotuki, Analysis of the potential
topical anti-inflammatory activity of Averrhoa carambola L. in mice, Evid. Based Complement.
Alternat. Med. 2011 (2011) 1-7.
[22] A. K. L. Rajahamsa, K. S. Deepak, T. K. V. K. Rao, A. V. R. P. Kumar, G. R. S. Reddy,
M. S. Potbhare, Multi-model confirmatory evaluation of anti-inflammatory, analgesic and
antioxidant activities of Puranjiva roxburghii wall, Int. J. Biomed. Advance Res. 12 (2013)
921-932.
[23] G. Coruzzi, C. Pozzoli, M. Adami, D. Grandi, N. Guido, R. Smits, I. de Esch, R. Leurs.
Strain-dependent effects of the histamine H4 receptor antagonist JNJ7777120 in a murine model
of acute skin inflammation. Exp. Dermatol. 21 (2012) 32-37.
[24] V. Kumar, A. Sharma, P. C. Sharma. Synthesis of some novel 2,5-disubstituted
thiazolidinones from a long chain fatty acid as possible anti-inflammatory, analgesic and
hydrogen peroxide scavenging agents. J. Enzyme Inhib. Med. Chem. 26 (2011) 198–203.
[25] T. Mosmann. Rapid colorimetric assay for cellular growth and survival: application to
proliferation and cytotoxicity assays. J. Immunol. Methods 65 (1983) 55-63.
[26] D. Scopel e Silva, C. C. Castro, S. F. Silva, V. Sant’anna, G. A. Vargas, M. Lima, G.
Fischer, A. Brandelli, A. S. da Motta, S. O. Hübner. Antiviral activity of a Bacillus sp. P34
peptide against pathogenic viruses of domestic animals, Braz. J. Microbiol. 45 (2014) 1089-
1094.
[27] O. Trott, A. Olson, AutoDock Vina: improving the speed and accuracy of docking with a
new scoring function, efficient optimization, and multithreading, J. Comput. Chem. 31 (2010)
455-461.

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2-Aryl-3-(2-morpholinoethyl)thiazolidin-4-ones: Synthesis, anti-inflammatory in
vivo, cytotoxicity in vitro and molecular docking studies.
Research Highlights:
ꢀ A serial of novel thiazolidinones were easily obtained by one-pot procedure
ꢀ Thiazolidinones showed anti-inflammatory activity in the ear edema model
ꢀ Four compounds had no toxic effect at 500 mg/ml in the Vero cell line
ꢀ One compound has six interaction with COX-2 enzyme according in silico docking
study