d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5

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Original Article

Synthesis of novel N-(3-chloro-4-flurophenyl)-

2-(5-((3-(4-hydroxy-2, 2-dimethyl-2,
3-dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazol-
4-yl) methylene)-4-oxo-2-thioxothiazolidin-3-yl)
acetamides having anti-inflammatory activity

K. Shyam Sunder a, Jayapal Maleraju b,*

a
Department of Organic Chemistry, Organic e II Division, IICT, Hyderabad 07, Andhra Pradesh, India
b
Department of Chemistry, Institute of Aeronautical Engineering, Hyderabad 07, Andhra Pradesh, India

article info abstract

Article history: Eight derivatives of N-(3-chloro-4-fluorophenyl)-2-(5-((3-(4-hydroxy-2, 2-dimethyl l-2,3-

Received 18 June 2013 dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazol-4-yl)methylene)-4-oxo-2-thioxothiazolidin-
Accepted 26 July 2013 3-yl) acetamide were synthesized by reacting pyrazole having substitutions at 1 and 3
positions (phenyl and benzofuran) with various substituted N-(3-chloro-4-fluorophenyl)-2-
(4-oxo-2-thioxothiazolidin-3-yl)[Link] series of reactions are depicted in
Keywords: following scheme. The chemical structures of these compounds were confirmed by means
Rhodanine acetamides of 1H NMR, IR and Mass spectra. The compounds were assayed for anti-inflammatory
1
H NMR activity.
IR Among the compounds 10a, 10b and 10d showed significant anti-inflammatory activity,
Mass spectra and anti-inflammatory 10c showed moderate anti-inflammatory activity.
activity Copyright ª 2013, JPR Solutions; Published by Reed Elsevier India Pvt. Ltd. All rights
reserved.

1. Introduction pharmacological activities like antibacterial,2 antifungal,3

anti-viral,4antimalarial,5 anti-tumour,6 anti-inflammator-
Rhodanines1 (2-thioxo-4-thiazolidinone) have become a very y,7anti-Alzhemers,6 anti-diabetic agents,8 insecticidal, anti-
interesting class of heterocyclic compounds since the intro- tubercular HCV NS3 protease, aldose reductase, activities
duction of EPALRESTAT into clinical use for the treatment of and also have been popular as small molecule inhibitors of
type-II diabetes mellitus and diabetic complications respec- numerous targets such as UGM inhibitors,9 UDP-N-acetyl
tively. Chemical modifications of these heterocycles muramate/L-alanine ligase inhibitors,10 MurG inhibitors,11
constantly result in compounds with a wide spectrum of belactamase inhibitors,12 RNA polymerase inhibitors.13

* Corresponding author. Department of Chemistry, Organic e II Division, Institute of Aeronautical Engineering, Hyderabad 07, Andhra
Pradesh, India.
E-mail address: mrjayapa007@[Link] (J. Maleraju).
0975-7619/$ e see front matter Copyright ª 2013, JPR Solutions; Published by Reed Elsevier India Pvt. Ltd. All rights reserved.
[Link]
 d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5 289

Pyrazole refers to the class of simple aromatic ring organic precipitate thus obtained was filtered and washed with 1:3
compounds of the heterocyclic series characterized by 5- (conc. HCl:H2O) solution, the crude product was dried and was
membered ring structure composed of three carbon atoms crystallized from dil. HCl.
and two nitrogen atoms in adjacent positions. Being so
composed and having pharmacological effects on humans 2.1.2. Preparation of 1-(2-hydroxy-4-(2-methyl-allyloxy)
they are classified as alkaloids, though they are rare in nature. phenyl) ethanone (3)
Pyrazoles are produced synthetically through the reaction of A mixture of resacetophenone (20.0 g, 0.131 mol), methallyl
a, b-unsaturated aldehydes with hydrazine and subsequent chloride (12.32 g, 0.131 mol), K2CO3 (36.15 g, 0.262 mol), cata-
dehydrogenation. Pyrazoles and their derivatives have been lytic amount of NaI & 90 ml of acetone was heated to reflux
proved to posses anti-tumour,14 anti-microbial,15 anti-viral,16 with stirring for 12 h. The reaction mixture was allowed to
leishmanicidal agents,17 antimycobacterial,18 immunosup- cool; inorganic salt was filtered and washed with acetone. The
pressant activity,19 LMW luteinizing hormone receptor ago- combined filtrate was concentrated and then water was added
nists,20 antiangiogenic,21 anticonvulsant,22 activities. and was filtered by suction, dried, weighed and was purified
by column chromatography.

2. Materials and methods 2.1.3. Preparation of 1-(2,4-dihydroxy-3-(2-methylallyl)

phenyl) ethanone (4)
All the required chemicals used were obtained from Aldrich 5 ml of diethyl aniline and 1 g of 1-(2-hydroxy-4-(2-methyl-
Chemicals and SD-Fine Chemicals. All the solvents used were allyloxy) phenyl) ethanone (0.485 mol) taken into refluxed for
of laboratory grade. Each reaction was monitored by TLC by 6 h while temperature maintaining below 220
C. The reaction
using appropriate solvent system, which was selected by trial mixture was then cooled and was poured into hexane. The
and error method. Precoated TLC plates (0.25 mm silica gel) precipitate obtained was filtered and dried. The crude product
were obtained from E. Merck. All the synthesized compounds was purified by column chromatography.
were purified by recrystallization. Melting points were deter-
mined on FishereJohns melting point apparatus and they 2.1.4. Preparation of 1-(4-Hydroxy-2, 2-dimethyl-2, 3-
were uncorrected. dihydrobenzofuran-5-yl) ethanone (5)
All 1H NMR spectrum were recorded on Avance 300 MHz Catalytic quantity of p-toluene sulphonic acid was added to
Bruker UX-NMR instrument and the samples were made in compound (4) and was heated to reflux in toluene for 6 h. The
CDCl3 and DMSO-d6 using tetra methyl silane (Me4Si) as the formation of product was monitored by TLC. The crude
internal standard. product was purified by column chromatography.
All the mass spectra were recorded on Thermo Finnigan
LCQ Ion Trap instrument and they were reported in m/z value 2.1.5. Preparation of 3-(4-hydroxy-2,2-dimethyl-2,3-dihydro-
as molecular ion peak. benzofuran-5-yl)-1-phenyl-1H-pyrazole-4-carbaldehyde (6)
IR spectra were recorded on Nexus 670 FTIR Thermo- To compound (5) (0.024 mol), phenyl hydrazine (0.024 mol)
Nicolet instrument by KBr disc method. was added and was kept for reflux for 12 h in presence of
The final compounds were synthesized as given below: ethanol. The reaction mixture was then concentrated and
crude product was filtered and to this crude product, cold
➢ Preparation of 3-(4-hydroxy-2,2-dimethyl-2,3-dihydro- solution of VilsmeiereHaack (VH) reagent i.e., DMF and POCl3
benzofuran-5-yl)-1-phenyl-1H-pyrazole-4-carbaldehyde was added. The mixture was stirred at 50e60
C for 5e6 h, was
(6) was carried out in 5 steps. cooled to room temperature, poured into cold water and
➢ Preparation of 2-substituted N-(4-chloro-3-fluorophenyl)- neutralized with saturated solution of sodium bicarbonate.
2-(4-oxo-2-thioxothiazolidin-3-yl) acetamide (9ae9h) The solid obtained was filtered and washed thoroughly with
was carried out in 2 steps. water. The obtained solid was purified by column
➢ The final compounds were synthesized by condensing 2- chromatography.
substituted N-(4-chloro-3-fluorophenyl)-2-(4-oxo-2-
thioxothiazolidin-3-yl) acetamide derivatives (9ae9h) 2.2. Preparation of 2-substituted N-(4-chloro-3-
with pyrazole aldehyde (6) fluorophenyl)-2-(4-oxo-2-thioxothiazolidin-3-yl) acetamide

2.2.1. Preparation of 2-substituted-2-(4-oxo-2-

2.1. Preparation of 3-(4-hydroxy-2,2-dimethyl-2,3- thioxothiazolidin-3-yl) acetic acid derivatives (8ae8h)
dihydro-benzofuran-5-yl)-1-phenyl-1H-pyrazole-4- Amino acid (10.0 g, 0.04 mol) was dissolved in KOH solution
carbaldehyde (6) [6 g (0.1 mol) in 50 ml water] and cooled in an ice bath. To this
4 g (0.05 mol) of carbon disulphide was added drop wise and
2.1.1. Preparation of resacetophenone (2) kept for stirring at room temperature overnight. To the
Resacetophenone preparation was carried out by heating a resultant brown solution, sodium salt of chloroacetic acid
mixture of ZnCl2 (33.0 g, 0.24 mol) with glacial acetic acid [prepared by adding 5 g (0.05 mol) of chloroacetic acid to the
(38.0 g, 0.63 mol) at 140
C. To this solution, resorcinol (22.0 g, solution of sodium carbonate (7 g (0.06 mol) of sodium car-
0.18 mol) was added in small portion keeping the temperature bonate in 25 ml water)] was added and stirred for 10 h. The
below 160
C. After 20 min the mixture was diluted with 1:1 reaction mixture was neutralized with 6 N HCl. The oily mass
(conc. HCl:H2O) solution and was cooled to 5
C. The obtained was extracted with ethyl acetate. The organic layer
290 d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5

was washed with brine solution and then with water and (9ae9h) required for condensation were prepared as depicted
dried over sodium sulphate and concentrated under reduced in Scheme 2:
pressure. The crude product obtained was purified by column The pyrazole aldehyde (6) was functionalized (Scheme 3) by
chromatography. condensation reaction with various substituted rhodanine
acetamides (9ae9h), in ethanol with catalytic mount of piper-
2.2.2. Preparation of 2-substituted N-(4-chloro-3- idine to give the corresponding derivatives (10ae10h). Table 1
fluorophenyl)-2-(4-oxo-2-thioxothiazolidin-3-yl) acetamide
(9ae9h) 3.2. Spectral data of synthesized compounds
2 g of (0.016 mol) 2-substituted-2-(4-oxo-2-thioxothiazolidin-
3-yl) acetic acid derivatives (8ae8h) was dissolved in dry DCM 3.2.1. 1-(2,4-Dihydroxyphenyl)-ethanone (2)
and to which added catalytic amount (0.1 ml) of dry DMF and IR (KBr) cm1: 3299.60 and 1606.31.
2 ml (0.015 mol) of oxallyl chloride under cold temperature EI-MS: 152 [86%, Mþ], 137 [100%], 109 [6%], 81 [21%], 69
(below 4
C). After the completion of the acid chloride for- [12%], 43 [13%], 41 [4%].
1
mation, the compound was concentrated. On the other hand H NMR [CDCl3, 300 MHz]: d 2.50 (s, 3H, CH3), 6.25 (d,
about 1.54 g (0.016 mol) of flouro chloro aniline was dissolved J ¼ 2.25 Hz, H-2), 6.31 (dd, J ¼ 2.2 Hz, J ¼ 8.8 Hz, H-6) and 7.54 (d,
in dry DCM and to which added drop wise acid chloride so- J ¼ 8.8 Hz, H-5).
lution and allowed the reaction for overnight. Then the com-
pound was extracted with ethyl acetate and the organic layer 3.2.2. 1-[2-Hydroxy-4-(2-methyl-allyloxy)-phenyl]-ethanone (3)
was washed with brine solution followed by water, dried over IR (KBr) cm1: 1636.99.
sodium sulphate and concentrated under reduced pressure. EI-MS: 206 [100%, Mþ.], 191 [82%], 163 [16%], 149 [14%], 137
The crude product obtained was purified by Column [33%], 123 [10%], 107 [8%], 95 [10%], 77 [12%], 69 [10%], 55 [60%],
chromatography. 40 [8%].
1
H NMR [CDCl3, 300 MHz]: d1.825 (s, 3H, eCH3), 2.532 (s, 3H,
2.3. General procedure for the synthesis of N-(3-chloro- eCH3), 4.439 (s, 2H, eCH2), 4.99 and 5.06 (s, 2H, eCH2), 6.358 (d,
4-fluorophenyl)-2-(5-((3-(4-hydroxy-2, 2-dimethyl-2,3- J ¼ 2.266 Hz, H-3), 6.408 (dd, J ¼ 3.022 Hz, J ¼ 2.2 Hz, H-5), 7.568
dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazol-4-yl) (d, J ¼ 9.06 Hz, H-6) and 12.59 (s, OH, D2O exchangeable).
methylene)-4-oxo-2-thioxothiazolidin-3-yl) acetamide
(10ae10h) 3.2.3. 1-[2,4-Dihydroxy-3-(2-methyl-allyl)-phenyl]-ethanone (4)
IR (KBr) cm1: 3167.72 and 1623.43.
10ae10h were prepared by dissolving an equimolar mixture EI-MS: 206 [100%, Mþ], 191 [100%], 173 [8%], 165 [14%] 149
of 2-substituted N-(4-chloro-3-fluorophenyl)-2-(4-oxo-2- [72%], 121 [10%], 85 [56%], 83 [85%], 65 [23%], 43 [82%], 41 [20%],
thioxothiazolidin-3-yl) acetamide (9ae9h) and 3-(4-hydroxy- 40 [8%].
1
2, 2-dimethyl-2, 3-dihydro-benzofuran-5-yl)-1-phenyl-1H- H NMR [CDCl3, 300 MHz]: d1.759 (s, 3H, CH3), 2.552 (s, 3H,
pyrazole-4-carbaldehyde (6) in ethanol. To these 3e4 drops of CH3), 3.475 (s, 2H, CH2), 4.92 (s, 2H, CH2), 5.95 (s, 1H, OH, D2O
piperidine was added and kept for stirring overnight at room exchangeable), 6.368 (d, J ¼ 8.684 Hz, H-5), 7.537 (d, J ¼ 8.684 Hz,
temperature. The formed product was then filtered and dried. H-6) and 13.052 (s, 1H, OH, D2O exchangeable).
The crude product was recrystallized from ethyl acetate and
hexane. 3.2.4. 1-(4-Hydroxy-2, 2-dimethyl-2, 3-dihydro-benzofuran-
5-yl)-ethanone (5)
IR (KBr) cm1: 1614.68.
EI-MS: 206 [86%, Mþ.], 191 [100%], 149 [16%], 137[10%], 83
3. Results and discussion [10%], 65 [8%], 43 [40%], 41 [10%].
1
H NMR [CDCl3, 300 MHz]: 1.495 (s, 6H, (CH3), 2.520 (s, 3H,
Synthesis of rhodanine acetamide derivatives incorporating CH3), 2.958 (s, 2H, CH2)), 6.244 (d, J ¼ 8.687 Hz, H-7), 7.529 (d,
pyrazole having substitutions at 1 and 3 positions (phenyl and J ¼ 8.687 Hz, H-6) and 12.633 (s, 1H, OH, D2O exchangeable).
benzofuran) has been planned firstly by synthesizing key in-
termediate benzofuran pyrazole aldehyde, which was then 3.2.5. 3-(4-Hydroxy-2, 2-dimethyl-2, 3-dihydrobenzofuran-5-
subsequently condensed with different rhodanine acetamide yl)-1-phenyl-1H-pyrazole-4-carbaldehyde (6)
derivatives. The series of reactions are depicted in following IR (KBr) cm1: 3423.18, 2923.43 and 1631.95.
schemes. EI-MS: 334 [90%, Mþ], 319 [60%], 291 [98%], 263 [10%], 191
[16%], 1117 [20%], 104 [24%], 91 [25%], 77 [100%], 65 [24%], 43
3.1. Synthesis of 3-(4-hydroxy-2, 2-dimethyl-2, 3- [50%], 41 [32%].
dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazole-4- ESI-MS: 333 [Mþ  1] Negative mode.
1
carbaldehyde (6) H NMR (CDCl3, 300 MHz, d): 1.529 (s, 6H, 2CH3), 3.043 (s, 2H,
CH2), 6.402 (d, J ¼ 7.365 Hz, H-7), 7.38 (d, J ¼ 7.365 Hz, H-6), 7.518
Starting from resorcinol, the pyrazole aldehyde (6) was syn- (t, 2H), 7.699 (t, 3H), 8.513 (s, 1H), 10.144 (s, OH, D2O
thesized in 5 steps as shown in Scheme 1. exchangeable) and 10.192 (s, CHO proton).
After the preparation of pyrazole aldehyde (6), the next
step in the synthesis is to functionalize aldehyde with 3.2.6. 2-(4-oxo-2-thioxothiazolidin-3-yl) acetic acid (8a)
substituted rhodanine acetamides. The rhodanine acetamides IR (KBr): 3404, 2927, 1704, 1631 and 1234 cm1
d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5 291

Scheme 1 e

Scheme 2
292 d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5

Scheme 3

EI-MS: Mass m/z ¼ 191,191 [100% Mþ], 173 [22%], 149 [12%], EI-MS: Mass m/z ¼ 267, 267 [65%, Mþ], 249 [62%], 192 [24%],
115 [25%], 90 [12%], 72 [84%], 64 [26%], 46 [72%]. 173 [32%], 148 [44%], 135 [20%], 117 [56%], 104 [90%], 89 [45%], 77
1
H NMR (300 MHz, DMSO-d6þCDCl3, d) 4.24 (s, 2H), 4.72 (s, [100%], 63 [21%], 46 [62%].
1
2H), 10.36 (s, OH, D2O exchangeable). H NMR (300 MHz, DMSO-d6þCDCl3, d): 4.22 (s, 2H), 6.67 (s,
1H), 7.21 (m, 3H), 7.5 (s, 2H), 10.26 (s, OH, D2O exchangeable).
3.2.7. 2-(4-oxo-2-thioxothiazolidin-3-yl)-2-phenylacetic acid
(8b) 3.2.8. N-(3-Chloro-4-fluorophenyl)-2-(4-oxo-2-
IR (KBr): 3359, 2926, 1770 and 1710 cm1. thioxothiazolidin-3-yl) acetamide (9a)
IR (KBr): 3253, 3062, 2924 and 1665 cm1
EI-MS: Mass m/z ¼ 318. 318 [12%, Mþ], 174 [100%], 158 [35%],
146 [60%], 118 [60%], 104 [16%], 72 [85%], 64 [8%], 59 [18%].
1
Table 1 e Physical data of synthesized compounds H NMR (300 MHz, DMSO-d6þCDCl3, d), 4.21 (s, 2H), 4.74 (s,
(10ae10h). 2H), 7.06 (t, 1H), 7.42 (m, 1H), 7.80 (dd, J ¼ 7.24 Hz, 1H), 10.24 (s,
S. no R Molecular Melting Yield (%) 1H).
weight point
3.2.9. N-(3-Chloro-4-fluorophenyl)-2-(4-oxo-2-
10a H 635.2 162 77
thioxothiazolidin-3-yl)-2-phenylacetamide (9b)
10b C6H5 711.12 158 81
10c CH2C6H5 725.25 164 83 IR (KBr): 3378, 2963 and 1241 cm1
10d CH(CH3)2 677.21 156 85 EI-MS: Mass m/z ¼ 394,394 [46%Mþ], 361 [83%].
10e CH2COOH 693.12 172 74 ESI-MS: 395 [M þ 1]þ
1
10f CH2CH2COOH 707.19 152 58 H NMR (300 MHz, DMSO-d6þCDCl3, d), 4.21 (s, 2H), 5.69 (m,
10g CHSH 679.20 166 85 1H), 7.23e7.26 (m, 3H), 7.33e7.39 (m, 3H), 7.48 (d, J ¼ 7.24, 1H)
10h CH2CH(CH3)2 691.23 165 82
7.85 (s, 1H), and 9.40 (s, 1H).
 d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5 293

1
3.2.10. N-(3-Chloro-4-fluorophenyl)-2-(4-oxo-2- H NMR (300 MHz, DMSO-d6, d): 1.48 (s, 6H), 2.98 (s, 2H), 4.48
thioxothiazolidin-3-yl)-3-phenylpropanamide (9c) (s, 2H), 6.32 (d, J ¼ 7.844 Hz, 1H), 7.01e7.06 (m, 1H), 7.14e7.18
IR (KBr): 3399, 2957, 1711, 1650 and 1209 cm1. (m, 1H), 7.34e7.36 (m, 1H), 7.46e7.50 (m, 2H), 7.66 (s, 1H),
ESI-MS : Mass: m/z ¼ 408, 409[Mþþ1]. 7.74e7.80 (m,2H), 7.84 (d, J ¼ 7.844 Hz, 2H),8.45 (s, 1H),9.40 (s,
1
H NMR (300 MHz, DMSO-d6þCDCl3, d), 2.51 (dd, 1H) and 10.23 (s, OH, D2O exchangeable).
J ¼ 16.43 Hz, 7.55 Hz, 2H), 4.21 (s, 2H), 5.69 (m, 1H), 7.23e7.26
(m, 3H), 7.33e7.39 (m, 3H), 7.48 (d, J ¼ 7.24, 1H) 7.85 (s, 1H), and 3.2.17. N-(3-Chloro-4-fluorophenyl)-2-(5-((3-(4-hydroxy-2, 2-
9.40 (s, 1H). dimethyl-2, 3-dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazol-4-
yl) methylene)-4-oxo-2-thioxothiazolidin-3-yl)-2-
3.2.11. N-(3-Chloro-4-fluorophenyl)-3-methyl-2-(4-oxo-2- phenylacetamide (10b)
thioxothiazolidin-3-yl) butanamide (9d) IR (KBr): 3424, 2965, 1691, 1635 and 1231 cm1
1
IR (KBr): 2970, 1716, 1635 and 1235 cm ESI-MS: Mass m/z ¼ 710, 711 [M þ 1]þ, 734 [M ¼ Na].
ESI-MS : Mass m/z ¼ 360, 361 [Mþ þ 1]. 1
H NMR (300 MHz, DMSO-d6, d): 1.48 (s, 6H), 2.98 (s, 2H), 5.78
1
H NMR (300 MHz, DMSO-d6þCDCl3, d) 0.77 (d, J ¼ 6.98 Hz, (s, 1H), 6.32 (d, J ¼ 9.18 Hz, 1H), 7.10e7.14 (m, 2H), 7.43e7.49 (m,
3H), 1.19 (d, J ¼ 6.98 Hz, 3H), 2.82e2.95 (m, 1H), 4.17 (s, 2H), 4.96 2H), 7.52e7.62 (m, 5H), 7.66e7.70 (m, 5H), 7.81 (d, J ¼ 7.347 Hz,
(d, 1H), 7.10 (t, 1H), 7.43e7.49 (m, 1H), 7.77 (dd, J ¼ 2.45 Hz, 1H), 1H), 8.45 (s, 1H), 9.40 (s, 1H) and 10.23 (s, OH, D2O
and 9.51 (s, 1H). exchangeable).

3.2.12. .4-(3-Chloro-4-fluorophenylamino)-4-oxo-3-(4-oxo-2- 3.2.18. N-(3-Chloro-4-fluorophenyl)-2-(5-((3-(4-hydroxy-2, 2-

thioxothiazolidin-3-yl) butanoic acid (9e) dimethyl-2, 3-dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazol-4-
IR (KBr): 3424, 2964, 1232 and 1598 cm1 yl) methylene)-4-oxo-2-thioxothiazolidin-3-yl)-3-
ESI-MS: Mass m/z ¼ 376,377[Mþ þ 1]. phenylpropanamide (10c)
1
H NMR (300 MHz, DMSO-d6þCDCl3, d) 1.26 (s, 2H), 2.01 (s, IR (KBr): 3424, 2958, and 1231 cm1
1H), 4.30 (s, 2H), 7.03e7.09 (t, 1H), 7.40e7.47 (m, 1H), 7.65 (s, 1H), ESI-MS: Mass m/z ¼ 724, 726 [M þ 1]þ
7.77e7.83 (dd, J ¼ 2.26 Hz, 2.42 Hz, 1H), 10.24 (s, 1H). 1
H NMR (300 MHz, DMSO-d6, d): 1.51 (s, 6H), 2.97 (s, 2H),
3.28e3.54 (m, 2H), 5.31 (t, 1H), 6.34 (d, J ¼ 7.48 Hz, 1H), 7.27e7.42
3.2.13. 5-(3-Chloro-4-fluorophenylamino)-5-oxo-4-(4-oxo-2- (m, 5H), 7.46e7.52 (m, 5H), 7.79e7.81 (m, 3H) 7.91 (d,
thioxothiazolidin-3-yl) pentanoic acid (9f) J ¼ 7.347 Hz, 1H), 8.45 (s, 2H), 9.45 (s, 1H) and 10.23 (s, OH, D2O
IR (KBr): 3442, 2895, 1691, 1635 and 1234 cm1 exchangeable).
ESI-MS: Mass m/z ¼ 390,391[Mþ þ 1].
1
H NMR (300 MHz, DMSO-d6þCDCl3, d): 1.26 (s, 2H), 2.01(s, 3.2.19. N-(3-Chloro-4-fluorophenyl)-2-(5-((3-(4-hydroxy-2, 2-
1H), 2.46 (s,2H), 4.30 (s, 2H), 7.03e7.09 (t, 1H), 7.40e7.47 (m, 1H), dimethyl-2, 3-dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazol-4-
7.65 (s, 1H),7.77e7.83 (dd, J ¼ 2.26 Hz, 2.42 Hz, 1H), 10.24 (s,1H). yl) methylene)-4-oxo-2-thioxothiazolidin-3-yl)-3-
methylbutanamide (10d)
3.2.14. N-(3-Chloro-4-fluorophenyl)-4-methyl-2-(4-oxo-2- IR (KBr): 3378, 2963, and 1241 cm1
thioxothiazolidin-3-yl) pentanamide (9g) ESI-MS: Mass m/z ¼ 676, 677 [M þ 1]þ
IR (KBr): 3348, 2958, 1672, 1635 and 1208 cm1 1
H NMR (300 MHz, DMSO-d6, d): 1.26 (m, 6H), 1.52 (d,
EI-MS: Mass m/z ¼ 374, 374 [65%, Mþ], 300 [12%], 241 [20%], J ¼ 10.78 Hz, 6H), 2.96e3.06 (m, 3H), 6.35 (s, 1H), 6.46 (d,
230 [80%], 187 [30%], 145 [35%], 134 [100%], 109 [25%], 85 [50%], J ¼ 7.84 Hz, 1H), 7.22e7.26 (t, 1H), 7.40e7.44 (t, 2H), 7.72 (d,
83 [75%], 69 [90%]. J ¼ 6.83 5H), 7.88 (s, 1H), 8.49 (s, 1H). and 10.13 (s, OH, D2O
1
H NMR (300 MHz, DMSO-d6þCDCl3, d) 0.99 (d, J ¼ 6.610 Hz, exchangeable).
6H), 1.46e1.57 (m, 1H), 1.96e2.07 (m, 1H), 2.27e2.38 (m, 1H),
3.97 (s, 2H), 5.64e5.70 (m, 1H), 7.05 (t, 1H), 7.29e7.35 (m, 1H),
and 7.55 (dd, J ¼ 2.644 Hz, 2H).
4. Screening for anti-inflammatory activity
3.2.15. N-(3-Chloro-4-fluorophenyl)-3-mercapto-2-(4-oxo-2-
thioxothiazolidin-3-yl) propanamide (9h) 4.1. Procurement and maintenance of animals
IR (KBr): 3424, 2965, 1691, 1635 and 1231 cm1
EI-MS: Mass m/z ¼ 364. Male Wistar rats weighing between 150 and 180 g were used
364 [100%, Mþ], 348 [82%], 225 [30%], 179 [72%], 145 [60%], 91 in the experiment and are obtained locally from Mahaveer
[75%], 57 [82%], 45 [100%], 41 [88%]. Enterprises, Hyderabad. The animals were acclimatized for
1
H NMR (300 MHz, DMSO-d6þCDCl3, d): 1.26 (s, 1SH), about 7 days prior to dosing. Cage numbers and individual
3.21e3.26 (m, 1H), 3.71 (s, 1H), 4.12 (s, 2H), 4.74 (s, 1H), 7.03e7.14 markings were made with a marker pen to identify the an-
(m, 2H), 7.62e7.69 (m, 1H), and 8.54 (s, 1H). imals. The animals were housed three per cage of same sex
in polypropylene cages provided with a bedding of paddy.
3.2.16. N-(3-Chloro-4-fluorophenyl)-2-(5-((3-(4-hydroxy-2, 2- Pellet chow feed standard diet under good management
dimethyl-2, 3-dihydrobenzofuran-5-yl)-1-phenyl-1H-pyrazol-4- conditions and water ad libitum was provided to the animals.
yl) methylene)-4-oxo-2-thioxothiazolidin-3-yl) acetamide (10a) They were maintained at standard environmental condi-
IR (KBr): 3252, 2924, 1665, and 1224 cm1 tions of temperature, relative humidity and dark/light
ESI-MS: Mass m/z ¼ 634, 635[M þ 1]þ. cycles.
294 d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5

4.2. Principle
Table 2 e Experiment protocol was done as follows.

Carrageenaneinduced paw oedema is the most commonly Group Compounds

used method in experimental pharmacology. Carrageenan is a Group 1 (control) 1% CMC
sulphated polysaccharide obtained from sea weed (Rhodo- Group 2 (standard) Ibuprofen (100 mg/kg)
phyceae), and by causing the release of histamine, 5-hy- Group 3 (10b)
droxytryptamine, bradykinin and prostaglandins it produces Group 4 (10c)
Group 5 (10d)
inflammation and oedema. The volume of oedema of rat hind
Group 6 (10e)
paw is measured prior and after inducing the inflammation Group 7 (10f)
using plethysmograph. Plethysmograph is a simple apparatus Group 8 (10g)
containing mercury in two glass arms A and B. The technique Group 9 (10h)
is based on mercury displacement .The mercury displacement
due to dipping of the paw can be read directly from a scale
attached to the mercury column or adjusting the mercury
level in the arm B to the original level by moving arm B up or was injected into dorsal region of sub-plantar surface of hind
down and noting the volume required to bring the level in paw of rat subcutaneously with the help of 26G needle.
both arms equal. The net oedema volume can be calculated by The paw volume was measured using the plethysmometer
subtracting paw volume before induction of oedema from the at 0, 30, 60,120,180, 240 min after the carrageenan challenge.
paw volume after the inflammation. The average value of oedema was calculated by taking the
average of each group at different hours. Percentage inhibition
4.3. Preparation of standard and test suspensions of oedema was calculated for each group with respect to the
control group. Table 2.
Suspensions of the standard drug, Ibuprofen and test com- Inflammation was expressed as the change in paw volume
pounds containing 20 mg/ml were prepared in 1% carboxy
Edema ¼ Tt  T0
methyl cellulose (1% CMC) solution.
Where T0 ¼ volume at 0 h, Tt ¼ Volume at t h
4.4. Route of administration of drugs
Percentage reduction ¼ ðV0  VT Þ=V0  100
The standard and test drugs were administered orally by an Where, VO ¼ volume of the paw of control at time ‘t’
oral feeding needle inserted into the posterior part of the VT ¼ volume of the paw of drug treated at time ‘t’.
pharynx. From the data obtained, the mean oedema volume and
percentage reduction in oedema was calculated.
4.5. Preparation of carrageenan suspension Screening for anti-inflammatory activity was performed
for all the substituted rhodanine acetamide derivatives at
1% suspension of carrageenan sodium salt was prepared by dose of 100 mg/kg weight in rats.
soaking 100 mg of carrageenan powder in 10 ml of saline (0.9% The drug was administered to the rats, and after 1 h, the
w/v NaCl) solution and homogeneous suspension was then inflammation inducing agent carrageenan was injected into
obtained by proper mixing. dorsal region of sub-plantar surface of hind paw. This allows
the evaluation of ability and potency of drug to protect from
4.6. Experimental procedure producing inflammation. The paw volumes were measured
after 0.5, 1, 2, 3 and 4 h of carrageenan administration.
In the present investigation, the anti-inflammatory activities Ibuprofen at a dose of 100 mg/kg was used as standard drug for
of 7 derivatives were tested by (chemical) carrageenan comparison. Table 3.
induced rat paw oedema method using Ibuprofen as standard.
Young adult male Wistar rats weighing 150e180 g were used,
which are acclimatized to the laboratory conditions and
maintained on standard laboratory rat feed and clean water.
Rats were fasted for 12 h prior to experiment, while allowing Table 3 e BAR diagram of percentage protection against
free access to water throughout the experiment. oedema formation.
The rats were divided into the groups of 6 animals each;
one group served as the control and received only the vehicle
i.e., the CMC. In the other groups, the test and standard drugs
were administered orally in a dose of 100 mg/kg of the body
weight (0.5 ml of 20 mg/ml drug solutions per 100 g body
weight). A mark was made on both the hind paws just beyond
the tibiotarsal junction, so that every time the paw is dipped in
the mercury column up to the marked level to ensure constant
paw volume.
After 1 h of administration of the test and standard sam-
ples, 0.1 ml of freshly prepared 1% carrageenan suspension
 d r u g i n v e n t i o n t o d a y 5 ( 2 0 1 3 ) 2 8 8 e2 9 5 295

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