6
K. R. A. Abdellatif et al.
J Enzyme Inhib Med Chem, Early Online: 1–7
(
DMSO-d ) ꢀ 2.50 (s, 3H, CH ), 7.04 (t, 1H, J ¼ 7.2 Hz, phenyl 1-Benzyl-5-methanesulfonyl-3-methyl-2-phenyl-1H-indole
6
3
ꢂ
H-4), 7.11–7.15 (m, 1H, indolyl H-5), 7.34–7.42 (m, 2H, indolyl (10f). Buff solid; Yield 42%; m.p. 235–237 C; IR (KBr) 3031
H-6, H-7), 7.45 (m, 7H, phenyl H-2, H-3, H-5, H-6, indolyl H-4 (CH aromatic) 2921, 2861 (CH aliphatic), 1343, 1182 (SO )
and benzoyl H-3, H-5), 7.68–7.95 (m, 3H, benzoyl H-2, H-4, H- cm ; H NMR (DMSO-d ) ꢀ 2.47 (s, 3H, CH ), 3.19 (s, 3H,
6
6
1
1
2
ꢀ1 1
3
1
3
); C NMR (DMSO-d ) ꢀ 9.63 (CH ), 113.82, 116.35, 119.78, SO CH ), 5.40 (s, 2H, CH ), 6.83 (d, 2H, J ¼ 7.6 Hz, benzyl H-2,
6
3
2
3
2
23.29, 124.89, 127.93, 128.27, 128.46, 129.61, 129.77, 130.10, H-6), 7.19–7.21 (m, 3H, benzyl H-3, H-4, H-5), 7.41–7.65 (m,
31.19, 132.43, 133.22, 135.37, 136.95, 169.47 (C¼O); EIMS 7H, phenyl protons and indole H-6, H-7), 8.18 (s, 1H, indole H-4);
+
+
13
(m/z) 312 (M + 1, 3.43%), 311 (M , 13.64%), 105 (C H O ,
7
C NMR (DMSO-d ) ꢀ 10.06 (CH ),45.03 (SO CH ), 47.35
6 3 2 3
5
1
Found: C, 85.13; H, 5.56; N, 4.38.
00%). Anal. Calcd for C H NO: C, 84.86; H, 5.50; N, 4.50. (CH ), 110.55, 111.59, 112.04, 119.38, 120.19, 127.66, 128.08,
2
2
2
17
128.23, 129.00, 129.12, 129.32, 130.73, 130.93, 132.16, 138.12,
1
+
38.72; EIMS (m/z) 376 (M + 1, 13.95%), 375 (M , 47.96%), 91
þ
(C7H , 100%). Anal. Calcd for C H NO S: C, 73.57; H, 5.64;
N, 3.73. Found: C, 73.42; H, 5.53; N, 3.81.
7
23 21
2
(
(
(
(
4-Chlorophenyl)-(3-methyl-2-phenyl-indol-1-yl)-methanone
ꢂ
10b). Buff solid; Yield 57%; m.p. 99–101 C; IR (KBr) 3061
ꢀ1 1
CH aromatic), 2992 (CH aliphatic), 1685 (C¼O) cm ; H NMR
Biological evaluation
DMSO-d ) ꢀ 2.40 (s, 3H, CH ), 7.10–7.34 (m, 8H, 5 phenyl
6
3
Animals
protons and indolyl H-5, H-6, H-7), 7.52–7.85 (m, 5H, indolyl H-
1
3
4
1
1
1
2
7
and 4 benzoyl protons); C NMR (DMSO-d ) ꢀ 9.64 (CH ),
6 3
Swiss albino mice (20–25 g) and Wistar rats (150–175 g) were
obtained from the National Research Center, Cairo, Egypt. They
were used throughout the study and were kept at controlled
14.05, 116.66, 119.81, 123.55, 125.15, 127.84, 128.51, 128.86,
30.21, 130.52, 131.95, 132.27, 134.19, 136.17, 136.89, 137.76,
+
68.63 (C¼O); EIMS (m/z) 347 (M + 2, 9.70%), 345 (M ,
ꢂ
conditions (temperature 23 ± 2 C, humidity 60 ± 10%) at a 12/
1
+
5.82%), 139 (C H ClO , 100%). Anal. Calcd for C H NO: C,
6.41; H, 4.66; 10.25; N, 4.05. Found: C, 76.35; H, 4.49; N, 4.23.
7
4
22 17
2 h light/dark cycle. All procedures relating to animal care and
treatments were conducted in accordance with protocols approved
by the Research Ethical Committee of Faculty of Pharmacy Beni-
Suef University (2014-Beni-Suef, Egypt).
2
-Benzyl-3-methyl-2-phenyl-1H-indole (10c). Yellow solid ;
0
1
Yield 49%; m.p. 110–112 C; IR (KBr) 3059, 3027 (CH
ꢂ
ꢀ
1 1
aromatic), 2923, 2859 (CH aliphatic) cm ; H NMR (DMSO-
COX-1/COX-2 inhibition colorimetric assay
d ) ꢀ 2.23 (s, 3H, CH ), 5.31 (s, 2H, CH ), 6.83 (d, 2H, J ¼ 6.8 Hz,
6
3
2
The in vitro inhibition of COX-1/COX-2 was measured using
colorimetric COX (ovine) Inhibitor Screening Assay Kit (Cayman
Chemical, Ann Arbor, MI) according to manufacturer’s instruc-
tions. This assay directly measures PGF2a that was produced by
stannous chloride reduction of COX-derived PGH2 by
enzyme immunoassay. All assays were conducted in triplicates
and IC50 values are the average of three determinations for each
compound.
benzyl H-2, H-6), 7.08–7.21 (m, 5H, benzyl H-3, H-4, H-5 and
indole H-5, H-6), 7.33 (d, 1H, J ¼ 7.6 Hz, indole H-7), 7.39–7.51
1
3
(
m, 5H, phenyl protons), 7.57 (d, 1H, J ¼ 7.6 Hz, indole H-4);
C
NMR (DMSO-d ) ꢀ 9.76 (CH ), 47.06 (CH ), 108.76, 110.89,
6
3
2
1
1
19.13, 119.65, 122.23, 126.43, 127.40, 128.52, 128.76, 128.87,
29.05, 130.71, 131.84, 136.81, 137.70, 138.87; EIMS (m/z) 298
+
þ
(M + 1, 12.57%), 297 (M , 53.35%), 91 (C7H , 100%). Anal.
Calcd for C H N: C, 88.85; H, 6.44; N, 4.71. Found: C, 88.93;
H, 6.56; N, 4.63.
7
2
2 19
Carrageenan-induced rat paw edema assay
(5-Methanesulfonyl-3-methyl-2-phenyl-indol-1-yl)-phenyl-metha-
none (10d). Buff solid; Yield 55%; m.p. 151–153 C; IR (KBr)
Rats were administered with vehicle, test compounds, indometh-
acin or celecoxib at a dose of 10 mg/kg by oral gavages.
Immediately thereafter, the rats received 100 mL of vehicle or
carrageenan (1% in saline) s.c. on the plantar surface of the left
hind paw under mild anesthesia, essentially, as reported before .
The paw edema was evaluated by measuring paw-volume changes
at 1, 3 and 6 h after carrageenan injection using a plethysmometer.
The right hind paw served as a reference of non-inflamed paw for
comparison. Results are expressed as paw-volume change (mL).
ꢂ
3
1
061 (CH aromatic), 2925, 2859 (CH aliphatic), 1677 (C¼O),
ꢀ
2 6
1
1
321, 1178 (SO ) cm ; H NMR (DMSO-d ) ꢀ 2.41 (s, 3H,
21
CH ), 3.19 (s, 3H, SO CH ), 7.21–7.72 (m, 11H, phenyl protons,
3
2
3
benzoyl protons and indolyl H-7), 7.76 (d, 1H, J ¼ 7.2 Hz, indolyl
1
3
H-6), 8.29 (s, 1H, indolyl H-4); C NMR (DMSO-d ) ꢀ 10.08
6
(CH ), 45.02 (SO CH ), 108.84, 112.03, 116.31, 119.60, 120.20,
3 2 3
1
1
0
27.33, 128.23, 128.64, 129.18, 129.31, 129.49, 130.35, 130.82,
31.60, 132.57, 138.79, 169.56 (C¼O); EIMS (m/z) 390 (M + 1,
+
þ
2
.27%), 389 (M , 0.93%), 285 (C16H15NSO , 100%). Anal. Calcd
Acetic acid-induced writhing test
for C H NO S: C, 70.93; H, 4.92; N, 3.60. Found: C, 80.13; H,
3
2
3
19
2
2
The writhing tests were carried as described before . Briefly,
mice were administered orally vehicle, test compounds, indo-
methacin or celecoxib at a dose of 10 mg/kg 30 min prior to
intraperitoneal administration of 0.7% v/v acetic acid solution
5
.04; N, 3.81.
(4-Chlorophenyl)-(5-methanesulfonyl-3-methyl-2-phenyl-indol-1-
yl)-methanone (10e). Buff solid; Yield 58%; m.p. 204–206 C;
ꢂ
(
10 mL/kg body weight). The number of writhes (i.e. abdominal
IR (KBr) 3047 (CH aromatic), 2943, 2914 (CH aliphatic), 1672
1
ꢀ
1
constriction followed by dorsiflexion and extension) occurring
during a 20-min period beginning 5 min after acetic acid injection
was measured. The results are expressed as the number of writhes
per 20-min period.
(
3
C¼O), 1317, 1175 (SO ) cm ; H NMR (DMSO-d ) ꢀ 2.33 (s,
2
6
H, CH ), 3.27 (s, 3H, SO CH ), 7.23–7.29 (m, 5H, phenyl
3 2 3
protons), 7.37 (d, 2H, J ¼ 8.0 Hz, benzoyl H-3, H-5), 7.56 (d, 2H,
J ¼ 8.0 Hz, benzoyl H-2, H-6), 7.81 (d, 2H, J ¼ 8.8 Hz, indolyl H-
7
), 7.86 (d, 2H, J ¼ 8.8 Hz, indolyl H-6), 8.29 (s, 1H, indolyl H-4);
C NMR (DMSO-d ) ꢀ 9.51 (CH ), 44.58 (SO CH ), 114.61,
1
3
Statistical analysis
6
3
2
3
1
1
16.61, 119.58, 123.30, 128.40, 128.69, 129.02, 130.24, 130.31, Comparisons among the groups were analyzed for statistical
31.48, 132.19, 133.35, 135.82, 138.42, 138.51, 139.00, 168.56 significance using the one-way ANOVA followed by Dunnett’s
+
(
(
C¼O); EIMS (m/z) 425 (M + 2, 4.52%), 423 (M , 11.15%), 139 test. Data were represented as mean ± standard deviation (SD).
+
C H ClO , 100%). Anal. Calcd for C H ClNO S: C, 65.17; H, Differences were considered significant at *p50.05, and
7
4
23 18
3
4
.28; N, 3.30. Found: C, 65.32; H, 4.41; N, 3.21.
**p50.01, respectively.