Research Article
Austin J Anal Pharm Chem. 2021; 8(2): 1136.
Ulcer Index and Anti-Inflammatory Testing of Some Benzimidazole Derivatives
Hussein AG¹*, Sakr HM², Mansour AM³ and Ayyad RR²
¹Ministry of Interior Medical Sector, Egypt
²Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Al-Azhar University, Egypt
³Department of Pharmacology, Faculty of Pharmacy, Al- Azhar University, Egypt
*Corresponding author: Ahmed Gaafar Hussein, Ministry of Interior Medical Sector, Egypt
Received: November 02, 2021; Accepted: November 26, 2021; Published: December 03, 2021
Abstract
In this work, we carry out the testing of some Benzimidazole derivatives as anti-inflammatory using Indomethacin 10mg/Kg, diclofenac sodium 7mg/ Kg, celecoxib 100mg/Kg and tested compounds of 200mg/Kg. The tested compounds showed anti-inflammatory activity in comparison with the standard reference drugs. In additionally, carrying out the testing of ulcer index for some testing compounds 600mg/Kg comparing with indomethacin 100mg/Kg, the testing revealed indomethacin causes ulcer in stomach of the testing animals, while the testing compounds no ulcerated the stomach of the testing animal.
Keywords: Benzimidazole; Anti-inflammatory; Ulcer index
Introduction
The Inflammation is a hard problem when somebody suffers from it. So the inflammation in any part of the body must be treated until no aggravate causing serious problems, one of these problems is the Tumor. So the inflammation when treated must take care in the choice of drug which used in the treatment.
Indomethacin used in treatment of many inflammations due to its activity as a powerful anti-inflammatory agent, but along time term of using it causes ulcerated stomach of the patient. Hance this work testing the some of benzimidazole derivatives 200mg/Kg as anti-inflammatory agent comparing with Indomethacin 10mg/Kg, diclofenac sodium 7mg/Kg, celecoxib 100mg/Kg as a reference drugs, and testing for some of benzimidazole 600mg/Kg comparing with Indomethacin 100mg/Kg
The chemical structure of heterocyclic compounds specially the structures which have benzo diazine and benzo diazole (phthalazine, Quinoxaline, Quinazoline and benzimidazole) have many of biological activities antibacterial, anticonvulsant, oral hypoglycemics, anticancer, anthelmintics, antiprotozoal, anti-inflammatory, analgesics, antioxidants, growth regulators, and other biological uses. so, we choose the benzimidazoles which is promising nucleus. it has analgesic activity like diclofenac and also has anticancer activity as doxirubsin i.e., six membered ring fused with five membered ring containing tow nitrogen like some compounds of anticancer, so we synthesized the following structure [1-25].
Chemistry
All of these compounds are reported [1] most of selected compounds (intermediate of final one) are tested as anti-inflammatory and carried out the ulcer index. Not all because of high cost of the biological studies.
The compounds prepared by the following Scheme 1.
Scheme 1:
Biological Studies
Experimental Animal for anti-inflammatory activity
Adult healthy Male Wister albino rats weighting between 140-160 gm were used for the study. The animals were housed in standard conditions (temperature 25±2°C with 55±5% relative humidity and a 12-hour light dark cycle). All animals had free access to water and normal diet. The study was approved by Institutional Animal Ethical Committee (IAEC) and was in accordance with the guideline of the Committee for the Purpose of Control and Supervision of Experimental Animal (CPCSEA).
In vivo anti-inflammatory activity
The initial paw volume of each rat was noted by the usage of Digital Vernier Caliper. One hundred fourty four animals were used in this study and divided into 24 groups (six animals per each).
Group-1 was served for saline injection in the right hind paw.
Group-2 was served for carrageenan injection in the right hind paw.
Group-3 was served for voltarin injection at a dose of 7mg/kg body weight.
Group-4 was received indomethacin at a dose of 10mg/kg body weight.
Group-5 was received celecoxib at a dose of 100mg/kg body weight.
Whereas groups 6-24 received the test samples at a dose of 200mg/kg body weight.
One hour post oral administration of test compounds at a dose of 200mg/kg, isotonic saline (0.1ml/paw), standards: diclofenac sodium (7mg/kg I.P), indomethacin (10mg/kg orally) and celecoxib (100mg/ kg orally) 1% w/v from carrageenan solution (0.1ml/paw) was injected subcutaneously into the plantar surface of the rat right leg hind paw.
The paw volume of the left legs; negative control for each animal was measured with the help of official Digital Vernier caliper during the time intervals of 1, 2, 3, 6, 12 and 24 h after carrageenan injection.
Percentage protection (or inhibition) was calculated by using the formula,
% protection = (1-Vt/Vc) X 100, where:
Vt is the mean increase in the paw volume in the test animals group,
Vc is the mean increase in the paw volume in the control group (in anti-inflammatory study).
Evaluation of edema, and percentage increase in paw volume in saline, carrageenan, standards and tested compounds (Table 1-23, Figure 1-24).
Figure 1: Saline group.
Figure 2: Carrageenan group.
Figure 3: Diclofenac sodium group.
Figure 4: Indomethacin group.
Figure 5: Celecoxib group.
Figure 6: IX treated group.
Figure 7: XVI treated group.
Figure 8: Vb treated group.
Figure 9: XV treated group.
Figure 10: XIVa treated group.
Figure 11: XIVb treated group.
Figure 12: VIIb treated group.
Figure 13: Vc treated group.
Figure 14: VIII treated group.
Figure 15: XIIIa treated group.
Figure 16: X treated group.
Figure 17: XIIIb treated group.
Figure 18: XIVc treated group.
Figure 19: XIIIc treated group.
Figure 20: Va treated group.
Figure 21: VIIa treated group.
Figure 22: IIIc treated group.
Figure 23: IIIb treated group.
Figure 24: VIIc treated group.
Rat no.
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.31
0.36
0.32
0.31
0.31
0.31
0.31
2
0.29
0.33
0.34
0.3
0.3
0.3
0.3
3
0.19
0.24
0.31
0.3
0.26
0.26
0.23
4
0.24
0.32
0.34
0.24
0.24
0.24
0.24
5
0.19
0.28
0.32
0.24
0.22
0.22
0.2
6
0.19
0.32
0.35
0.29
0.26
0.23
0.22
Total
141
185
198
168
159
156
150
Mean
23.5
30.8
33
28
26.5
26
25
% of S.D
------
31.2
40.4
19.1
12.7
10.6
6.3
Table 1: Saline group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.3
0.6
0.85
0.85
0.89
0.94
0.6
2
0.25
0.75
1.12
0.91
0.8
0.85
0.63
3
0.24
0.8
0.99
0.9
0.71
0.88
0.65
4
0.32
0.82
0.98
0.95
0.91
0.96
0.74
5
0.26
0.81
0.96
0.9
0.85
0.88
0.58
6
0.27
0.82
0.97
0.92
0.84
0.88
0.61
Total
1.64
2.6
5.87
5.43
5
5.39
3.81
Mean
0.273
0.766
0.978
0.905
0.833
0.898
0.635
% of S.D
-------
58.5
258
231
204
228
132
Table 2: Carrageenan group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.3
0.42
0.5
0.6
0.7
0.77
0.62
2
0.29
0.35
0.4
0.5
0.54
0.52
0.44
3
0.24
0.4
0.45
0.4
0.38
0.49
0.45
4
0.25
0.38
0.41
0.44
0.52
0.61
0.55
5
0.24
0.41
0.42
0.46
0.53
0.61
0.44
6
0.26
0.4
0.44
0.48
0.56
0.63
0.43
Total
1.58
2.36
2.62
2.88
3.23
3.63
2.93
Mean
0.263
0.393
0.436
0.48
0.538
0.605
0.488
% of S.D
------
49.3
65.8
82.3
104.4
129.7
85.4
Table 3: Diclofenac sodium group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.48
0.54
0.6
0.65
0.59
0.44
2
0.28
0.33
0.36
0.4
0.41
0.46
0.3
3
0.24
0.35
0.38
0.54
0.6
0.48
0.35
4
0.3
0.32
0.35
0.36
0.39
0.51
0.4
5
0.28
0.34
0.37
0.38
0.4
0.52
0.41
6
0.27
0.35
0.39
0.4
0.42
0.54
0.42
Total
1.63
2.17
2.39
2.68
2.87
3.1
2.32
Mean
0.271
0.361
0.398
0.446
0.478
0.516
0.386
% of S.D
------
33.1
46.6
64.4
76
90.2
42.3
Table 4: Indomethacin group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.28
0.47
0.56
0.49
0.4
0.35
0.35
2
0.26
0.43
0.54
0.49
0.49
0.39
0.38
3
0.27
0.43
0.55
0.45
0.35
0.35
0.35
4
0.28
0.51
0.58
0.49
0.39
0.33
0.33
5
0.24
0.5
0.56
0.46
0.4
0.4
0.34
6
0.25
0.5
0.58
0.45
0.38
0.38
0.36
Total
1.6
2.84
3.37
2.83
2.41
2.2
2.11
Mean
0.266
0.47
0.56
0.47
0.4
0.36
0.35
% of S.D
------
77
111
77
51
37
32
Table 5: Celecoxib group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.28
0.6
0.7
0.8
0.9
0.75
0.6
2
0.23
0.55
0.7
0.84
0.84
0.76
0.66
3
0.26
0.6
0.75
0.8
0.75
0.7
0.64
4
0.24
0.56
0.68
0.74
0.77
0.75
0.54
5
0.27
0.55
0.7
0.75
0.7
0.7
0.46
6
0.29
0.5
0.65
0.68
0.74
0.6
0.44
Total
1.57
3.36
4.19
4.61
4.7
4.26
3.34
Mean
0.262
0.56
0.7
0.77
0.78
0.71
0.56
% of S.D
------
114
167
193
199
171
112
Table 6: IX treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.7
0.83
0.8
0.665
0.6
0.518
2
0.26
0.68
0.8
0.87
0.787
0.7
0.646
3
0.28
0.7
0.93
0.95
0.92
0.85
0.77
4
0.27
0.7
0.91
0.92
0.71
0.65
0.565
5
0.28
0.65
0.9
0.85
0.7
0.63
0.55
6
0.27
0.66
0.95
0.88
0.72
0.68
0.65
Total
1.62
4.09
5.32
5.27
4.5
4.11
3.68
Mean
0.27
0.68
0.88
0.87
0.75
0.68
0.61
% of S.D
------
147
220
216
172
147
121
Table 7: XVI treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.27
0.6
0.7
0.9
0.96
0.8
0.74
2
0.28
0.6
0.7
0.92
0.97
0.8
0.7
3
0.24
0.61
0.66
0.92
0.92
0.7
0.59
4
0.27
0.6
0.68
0.78
0.8
0.68
0.6
5
0.28
0.56
0.7
0.8
0.8
0.67
0.6
6
0.26
0.66
0.76
0.82
0.85
0.7
0.61
Total
1.6
3.63
4.2
5.14
5.27
4.35
4.04
Mean
26.6
0.605
0.7
0.856
0.878
0.725
0.673
% of S.D
------
127
163
222
230
177
153
Table 8: Vb treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.65
0.83
0.8
0.665
0.6
0.618
2
0.26
0.68
0.8
0.87
0.787
0.7
0.646
3
0.28
0.63
0.82
0.95
0.82
0.75
0.67
4
0.29
0.65
0.81
0.92
0.71
0.62
0.565
5
0.29
0.6
0.8
0.85
0.7
0.63
0.5
6
0.27
0.56
0.85
0.88
0.72
0.68
0.65
Total
1.65
3.77
4.91
5.27
4.39
3.98
3.63
Mean
0.275
0.63
0.82
0.88
0.73
0.66
0.6
% of S.D
------
129
198
220
165
140
118
Table 9: XV treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.65
0.73
0.88
0.98
0.9
0.84
2
0.27
0.62
0.7
0.9
0.98
0.55
0.51
3
0.3
0.61
0.66
0.9
0.98
0.64
0.6
4
0.3
0.54
0.68
0.83
0.9
0.8
0.74
5
0.28
0.52
0.72
0.75
0.8
0.7
0.54
6
0.3
0.56
0.68
0.88
0.93
0.7
0.68
Total
1.71
3.5
4.17
5.14
5.57
4.29
3.91
Mean
0.285
0.58
0.69
0.86
0.93
0.72
0.65
% of S.D
------
105
144
202
226
151
129
Table 10: XIVa treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.28
0.5
0.79
0.9
0.89
0.8
0.65
2
0.28
0.68
0.74
0.89
0.93
0.83
0.72
3
0.25
0.6
0.7
0.7
0.7
0.7
0.6
4
0.28
0.55
0.88
0.78
0.79
0.75
0.63
5
0.27
0.65
0.8
0.85
0.8
0.74
0.7
6
0.26
0.58
0.75
0.88
0.84
0.76
0.72
Total
1.62
3.56
4.66
5
4.95
4.58
4.02
Mean
0.27
0.59
0.78
0.83
0.82
0.76
0.67
% of S.D
------
120
187
208
206
182
148
Table 11: XIVb treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.66
0.75
0.9
0.9
0.7
0.6
2
0.26
0.65
0.8
0.99
0.84
0.72
0.67
3
0.29
0.55
0.85
0.99
0.9
0.8
0.71
4
0.28
0.66
0.83
0.9
0.9
0.75
0.62
5
0.28
0.5
0.9
0.95
0.8
0.72
0. 600
6
0.26
0.6
0.85
0.88
0.78
0.7
0.64
Total
1.63
3.63
4.98
5.62
5.12
4.39
3.84
Mean
0.271
0.61
0.83
0.94
0.85
0.73
0.64
% of S.D
------
123
206
246
215
170
136
Table 12: VIIb treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.55
0.7
0.84
0.9
0.8
0.7
2
0.27
0.53
0.66
0.82
0.787
0.78
0.72
3
0.28
0.6
0.76
0.95
0.82
0.82
0.77
4
0.27
0.5
0.68
0.92
0.8
0.78
0.565
5
0.28
0.55
0.7
0.85
0.8
0.76
0.55
6
0.26
0.5
0.67
0.88
0.82
0.8
0.65
Total
1.62
3.23
4.17
5.26
4.92
4.74
3.95
Mean
0.27
0.54
0.7
0.88
0.82
0.79
0.66
% of S.D
------
100
159
226
203
193
144
Table 13: Vc treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.28
0.6
0.79
0.9
0.8
0.75
0.7
2
0.23
0.68
0.84
0.89
0.9
0.83
0.72
3
0.26
0.6
0.75
0.9
0.75
0.7
0.6
4
0.24
0.65
0.88
0.98
0.79
0.75
0.63
5
0.27
0.55
0.7
0.85
0.7
0.74
0.6
6
0.29
0.58
0.75
0.88
0.64
0.7
0.58
Total
1.57
3.66
4.71
5.4
4.58
4.47
3.83
Mean
0.261
0.61
0.785
0.9
0.76
0.74
0.64
% of S.D
------
100
201
245
191
184
145
Table 14: VIII treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.28
0.7
0.83
0.99
0.95
0.85
0.57
2
0.26
0.6
0.8
0.97
0.84
0.75
0.68
3
0.25
0.63
0.76
0.8
0.85
0.7
0.57
4
0.24
0.65
0.78
0.83
0.83
0.72
0.6
5
0.29
0.6
0.72
0.77
0.8
0.67
0.6
6
0.28
0.56
0.6
0.67
0.72
0.6
0.58
Total
1.6
3.74
4.49
5.03
4.99
4.29
3.6
Mean
0.266
0.62
0.75
0.838
0.83
0.715
0.6
% of S.D
------
133
182
212
212
166
77
Table 15: XIIIa treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.28
0.7
0.83
0.88
0.7
0.65
0.57
2
0.26
0.6
0.7
0.8
0.71
0.55
0.48
3
0.27
0.63
0.66
0.8
0.66
0.54
0.46
4
0.26
0.56
0.78
0.83
0.7
0.6
0.54
5
0.29
0.58
0.72
0.75
0.7
0.6
0.54
6
0.28
0.56
0.58
0.66
0.6
0.6
0.58
Total
1.64
3.63
4.27
4.72
4.07
3.54
3.17
Mean
0.273
0.6
0.71
0.78
0.68
0.59
0.53
% of S.D
------
121
160
188
148
116
100
Table 16: X treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.5
0.6
0.74
0.59
0.5
0.36
2
0.27
0.53
0.66
0.82
0.77
0.7
0.52
3
0.28
0.6
0.76
0.85
0.72
0.7
0.6
4
0.27
0.6
0.68
0.82
0.7
0.68
0.55
5
0.28
0.55
0.7
0.8
0.72
0.6
0.55
6
0.26
0.55
0.66
0.82
0.7
0.7
0.6
Total
1.62
3.33
4.06
4.85
4.2
3.88
3.18
Mean
0.27
0.55
0.68
0.81
0.7
0.65
0.53
% of S.D
------
98
151
199
159
140
96
Table 17: XIIIb treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.28
0.7
0.83
0.88
0.7
0.65
0.57
2
0.26
0.6
0.7
0.8
0.71
0.55
0.48
3
0.27
0.63
0.66
0.8
0.66
0.54
0.46
4
0.26
0.56
0.78
0.83
0.7
0.6
0.54
5
0.29
0.58
0.72
0.75
0.7
0.6
0.54
6
0.28
0.56
0.58
0.66
0.6
0.6
0.58
Total
1.64
3.63
4.27
4.72
4.07
3.54
3.17
Mean
0.273
0.6
0.71
0.78
0.68
0.59
0.53
% of S.D
------
121
160
188
148
116
100
Table 18: XIVc treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.5
0.6
0.72
0.57
0.48
0.34
2
0.27
0.53
0.66
0.8
0.75
0.68
0.5
3
0.28
0.58
0.78
0.83
0.7
0.68
0.58
4
0.27
0.62
0.7
0.8
0.68
0.66
0.53
5
0.26
0.55
0.7
0.78
0.7
0.58
0.53
6
0.28
0.55
0.66
0.8
0.68
0.68
0.58
Total
1.62
3.33
4.06
4.73
4.08
3.76
3.06
Mean
0.27
0.55
0.68
0.78
0.68
0.62
0.51
% of S.D
------
98
151
191
152
132
89
Table 19: XIIIc treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.6
0.78
0.8
0.69
0.6
0.5
2
0.24
0.55
0.8
0.8
0.63
0.56
0.54
3
0.28
0.55
0.85
0.82
0.44
0.4
0.38
4
0.27
0.55
0.78
0.74
0.45
0.45
0.34
5
0.28
0.5
0.8
0.8
0.5
0.44
0.36
6
0.26
0.55
0.85
0.85
0.62
0.48
0.35
Total
1.59
3.3
4.86
4.81
3.33
2.93
2.47
Mean
0.265
0.55
0.81
0.8
0.555
0.49
0.41
% of S.D
------
108
205
202
109
84
55
Table 20: Va treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.27
0.5
0.5
0.52
0.45
0.4
0.29
2
0.35
0.63
0.76
0.87
0.62
0.55
0.4
3
0.24
0.61
0.76
0.8
0.58
0.5
0.41
4
0.27
0.54
0.68
0.85
0.55
0.5
0.38
5
0.28
0.55
0.75
0.78
0.6
0.55
0.42
6
0.26
0.56
0.76
0.85
0.65
0.6
0.48
Total
1.68
3.39
4.21
4.67
3.45
3.1
2.38
Mean
0.28
0.565
0.7
0.78
0.575
0.52
0.396
% of S.D
------
102
151
179
105
85
42
Table 21: VIIa treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.3
0.55
0.5
0.6
0.55
0.45
0.4
2
0.36
0.65
0.76
0.87
0.62
0.55
0.4
3
0.24
0.61
0.8
0.8
0.58
0.5
0.41
4
0.27
0.54
0.68
0.85
0.65
0.5
0.38
5
0.27
0.55
0.75
0.78
0.6
0.55
0.42
6
0.26
0.56
0.76
0.85
0.65
0.6
0.48
Total
1.7
3.46
4.25
4.75
3.65
3.15
2.49
Mean
0.283
0.58
0.71
0.792
0.61
0.525
0.415
% of S.D
------
103
150
179
115
86
47
Table 22: IIIc treated group.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.65
0.73
0.88
0.53
0.4
0.34
2
0.27
0.62
0.7
0.71
0.62
0.55
0.49
3
0.3
0.61
0.66
0.71
0.53
0.44
0.34
4
0.3
0.74
0.78
0.94
0.51
0.5
0.44
5
0.28
0.52
0.72
0.81
0.6
0.55
0.4
6
0.3
0.66
0.8
0.88
0.63
0.6
0.46
Total
1.71
3.8
4.39
4.93
3.42
3.04
2.47
Mean
0.285
0.633
0.73
0.822
0.57
0.506
0.41
% of S.D
------
122
157
188
100
78
44
Table 23: IIIb treated group.
The anti-inflammatory effects of saline, diclofenac sodium, indomethacin, Celecoxib and tested compounds administration in comparison to carrageenan injected rats
Data in Table 24 showed that injection of 0.1ml saline in right hind paw (S.C) caused no significant increase in right paw volume by about (31.2%, 40.4%, 19.1% 12.7% 10.6% and 6.3% after 1, 2, 3, 6, 12 and 24 h, respectively, when compared to volume of left leg, while injection of 0.1ml carrageenan in right hind paw (S.C) in a dose of 1% w/v caused a significant increase in right paw volume by about (58.5%, 258%, 231% 204% 228% and 132% after 1, 2, 3, 6, 12 and 24 h, respectively, when compared to volume of left leg, while pretreatment with diclofenac sodium (I.P) in a dose of 7mg/kg caused a significant reduction in right paw volume by (49.3%, 65.8%, 82.3%, 104.4%, 129.7% and 85.4%), after 1, 2, 3, 6, 12 and 24 h, respectively,. when compared to volume of left leg The pretreatment with indomethacin (orally) in a dose of 10mg/kg caused a significant reduction in right paw volume by, (33.1%, 46.4%, 64.4%, 76.0%, 90.2% and 42.3%), after 1, 2, 3, 6, 12 and 24 h, respectively, when compared to volume of left leg. While pretreatment with Celecoxib (orally) in a dose of 100mg/kg caused a significant reduction in right paw volume by (77%, 111%, 77%, 51%, 37% and 32%), after 1, 2, 3, 6, 12 and 24 h, respectively, when compared to volume of left leg. The compounds XIIIa, X, XIIIb, IX, VIII, Vc, VIIb, XVI, XV, XIVb and XIVa at a dose of (200mg/kg orally) shows no significant reduction in right hind paw volume after 1, 2, 3, 6, 12 and 24 h, respectively, when compared to volume of left leg .taken in consideration that this compounds had no activity when compared to three standards (diclofenac sodium, indomethacin and Celecoxib). The compounds Vb, and Va) at a dose of (200mg/kg orally) were failed to finish the experiment and most of animals were died and the remaining number not sufficient to carry out the statistics, taken in consideration that this compounds had no activity when compared to three standards (diclofenac sodium, indomethacin and Celecoxib). The use of tested compounds (XIVc and XIIIc) at (200mg/kg orally) had mild reduction in right paw volume when compared to standard treated groups after 1, 2, 3, 6, 12 and 24 h, respectively, The use of tested compound (IIIb) at (200mg/ kg orally) had mild to moderate reduction in right paw volume after 1, 2, 3, 6, 12 and 24 h, respectively, when compared to standard treated groups. The use of tested compounds (VIIa, IIIc and VIIc) at (200mg/ kg orally) had a remarkable promising reduction in right paw volume after 1, 2, 3, 6, 12 and 24 h, respectively, when compared to standard treated groups.
Figure 25: Normal stomach.
Figure 26: Indomethacin treated group.
Figure 27: Compound VIIc.
Figure 28: Compound VIII.
Rat no
Left leg
1 h
2 h
3 h
6 h
12 h
24 h
1
0.26
0.65
0.7
0.88
0.63
0.5
0.3
2
0.27
0.6
0.7
0.81
0.62
0.55
0.49
3
0.24
0.61
0.66
0.77
0.63
0.54
0.34
4
0.25
0.7
0.78
0.9
0.81
0.6
0.44
5
0.28
0.5
0.76
0.86
0.76
0.55
0.4
6
0.27
0.6
0.8
0.88
0.63
0.6
0.46
Total
1.57
3.66
4.4
5.1
4.08
3.34
2.43
Mean
0.262
0.61
0.73
0.85
0.68
0.556
0.405
% of S.D
------
133
180
224
160
112
55
Table 24: VIIc treated group.
Discussion
The present study showed that, the selected dose (200 mg/kg) of tested compounds were evaluated for their in-vivo anti-inflammatory activity and compared to diclofenac sodium, indomethacin and Celecoxib as a references were measured before and 1, 2, 3, 6, 12, and 24 h, after carrageenan injection. Percent of the oedema inhibition was calculated as a regard to carrageenan control group and potency was calculated as a regard to the percentage of the change of the diclofenac sodium, indomethacin and Celecoxib and tested compounds, it was observed that the 200mg/kg dose of compounds (XIIIa, X, XIIIb, IX, VIII, Vc, VIIb, XVI, XV, XIVb and XIVa) had shown no antiinflammatory activity at all-time points. The compounds (Vb and Va) at a dose of (200mg/kg orally) were failed to finish the experiment. It was observed that the 200mg/kg dose of compounds (XIVc and XIIIc) had shown mild anti-inflammatory activity at all-time points, while tested compounds, it was observed that the 200mg/kg dose of compound (IIIb) had shown mild to moderate anti-inflammatory activity at all-time points, it was observed that the 200mg/kg dose of compounds (VIIa, IIIc and VIIc) had shown highest antiinflammatory activity at all-time points.
Ulcer Index Experiment
Materials and methods
Drugs: Fresh solution of indomethacin 25 mg/kg was purchased from (El-Nile Co. for Pharmaceuticals and chemical industries, Cairo, Egypt), VIIc, VIII, IX, X, Va, Vb, XIVa, XIVb, XIVc, XIIIa and XIIIb the tested compounds were dissolved in DMSO while indomethacin in sterile water was prepared.
Animals: Seventy-eight female Sprague Dawly rats weighing between 150 ± 10 gm were used for the study. Prior to the experiments, the rats were kept in the animal house for one week for acclimatization in rat cages and were given standard rat feed with water ad libitum.
The animals were kept fasting for 24 hours before carrying out the experiments.
Experimental procedures
Rats were divided into 13 groups of 6 each:
Group I: Control (1ml/kg DMSO solvent orally).
Group II: Indomethacin (25mg/kg orally).
Group III: VIIc (600mg/kg orally in DMSO solvent).
Group IV: VIII (600mg/kg orally in DMSO solvent).
Group V: IX (600mg/kg orally in DMSO solvent).
Group VI: X (600mg/kg orally in DMSO solvent).
Group VII: Va (600mg/kg orally in DMSO solvent).
Group VIII: Vb (600mg/kg orally in DMSO solvent).
Group IX: XIVa (600mg/kg orally in DMSO solvent).
Group X: XIVb (600mg/kg orally in DMSO solvent).
Group XI: XIVc, (600mg/kg orally in DMSO solvent).
Group XII: XIIIa (600mg/kg orally in DMSO solvent).
Group XIII: XIIIb (600mg/kg orally in DMSO solvent).
The rats were sacrificed by cervical dislocation 3 hours after indomethacin and tested compounds administration. The abdomen was dissected to retrieve stomach, analyzed for ulcer index. With small nick, fundus of stomach was perforated on greater curvature of stomach. The greater curvature of stomach was opened. Gastric mucosa was observed under magnifying glass to calculate the ulcer index.
Measurement of gastric lesions
Measurement of gastric ulcerations following their induction is done by first dissecting the stomach along its greater curvature and fixing on a board or transparent glass.
Examination can be carried out macroscopically with a hand lens and by tracing on a transparent paper after which the transparent paper is placed onto a graph sheet and sizes of ulcers are measured.
According to the method by Kulkarni, the ulcer index can be measured or registered using the following scores involving the number and severity of ulcers:
0.0 = normal colored stomach,
0.5 = red coloration,
1.0 = spot ulcers,
1.5 = hemorrhagic streaks,
2.0 = ulcers with area >3 but ≤5mm²,
Statistical analysis
All the data are expressed as mean ± standard error of mean. One-way analysis of variance (ANOVA) test (SPSS Version no. 15) unpaired one tailed ‘t’ test was also used considering P ≤0.05 to be statistically significant (Figure 29-36).
Figure 29: Compound XIVC.
Figure 30: Compound XII.
Figure 31: Compound XIVa.
Figure 32: Compound XIVb.
Figure 33: Effects of indomethacin and VIIc, VIII, IX, X1, Va, Vb, XIVa, XIVb,
XIVc, XIII1 and XIII2, on gastric mucosa of rats compared to control group.
Data are expressed as means ± SEM of six rats per group.
Indomethacin (25mg/kg), VIIc, VIII, IX, X1, Va, Vb, XIVa, XIVb, XIVc, XIII1 and
XIII2 (600mg/kg) were given orally single dose.
aSignificantly different from control group.
bSignificantly different from indomethacin-administered group using one-way
analysis of variance (ANOVA) followed by Tukey-Kramer test for multiple
comparison at P ≤0.05.
Effects of indomethacin and VIIc, VIII, IX, X, Va, Vb, XIVa, XIVb, XIVc, XIIIa and XIIIb, on gastric mucosa of rats compared to control group
The results in Figure 29 illustrate that the effects of indomethacin and VIIc, VIII, IX, X1, Va, Vb, XIVa, XIVb, XIVc, XIII1 and XIII2, on incidence of ulcer in rats compared to control group.
Apparently, oral administration with indomethacin in a dose of 25 mg/kg body weight significantly increase the incidence of ulcer index (100%) compared to the control group. Similarly, oral administration with compounds VIIc, XIVa and XIVc significant decrease the incidence of ulcer index to 50% in comparison with indomethacin-administered group, while compounds, VIII, Va, Vb, and XIVb significant decrease the incidence of ulcer index to 75% in comparison with indomethacin-administered group. In addition, the compound XIIIa showed non-significant reduction in ulcer index in comparison with indomethacin-administered group, finally the compound XIIIb induced ulcer index greater than that occurs with indomethacin treated group. Unfortunately, the animals that treated with compounds IX and X were died during the carried out of the experiments (Figure 37).
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