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SYNTHESIS, ANTI-INFAMMATORY AND ANALGESIC EVALUATION
OF SOME 2-AMINO-5-SELENOTHIAZOLES
Moustafa A. Gouda a,c, , Yousery El-Sayed Sherif b,c, and Mohamed S. Elsherbinid
a
b
Chemistry Department, Faculty of Science, Mansoura University, Mansoura, 35516, Egypt
Department of Clinical Pharmacology, Faculty of Medicine, Mansoura University, Mansoura,
Egypt
c
Chemistry Department, Faculty of Science and Arts, Ulla, Taibah University, KSA
d
Chemistry Department, Faculty of Science, Al Jouf University, Sakaka, KSA
Email: dr_mostafa_chem@yahoo.com
Supplemental Materials
Biological evaluation
The anti-inflammatory activity and Analgesic effect of the synthesized compounds were
evaluated by collagen II-adjuvant induced paw edema test in rats. As shown in Table S 1,
Meloxicam was able to modify the rheumatoid index (RI), pain tolerance, and pain scoring
positively.
Measurement of joint inflammation
The anti-inflammatory activities of the synthesized compounds were evaluated by collagen II–
adjuvant induced paw edema test in rats. The anti-inflammatory activity data (Table S 1)
indicated that most of the tested compounds protected rats from paw edema-induced
inflammation. Compounds 3, 4, 7, 16, 19, 20, 21 and 22 showed similar and higher antiinflammatory activity than Meloxicam. Compound 7 was found as the most effective compound
in reduction of rheumatoid index versus all other compounds. On the other hand, the antiinflammatory activity data (the thickness of edema) indicated that all the tested compounds
showed better results than Meloxicam (Table S 1).
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Table S 1: Influence of Meloxicam and twelve compounds in analgesic effect, and inflammation
in adjuvant induced rheumatoid arthritis model in rats (M± S.E), n number of rats = 6
Group
Dose
(mg
Inflammation
-1
200 g ) [29]
--
--
Rheumatoid
index
Edema
thickness
Pain
tolerance
Pain scoring
Negative
control
[SCMC]
Solvent
1.0±0.00
2.3±0.22
14.33±2.02
1±0.
Positive
control
[SCMC]
Solvent
3.67±0.22*
5.0±0.36*
1.33±0.21*
3.67±0. 21*
Meloxicam
0.36
2.67±0.22*
4.3±0.42* θ
3.33±0.84*
2.67±0. 21*θ
1
1.10
3.0±0.0
3.67±0.28*
2.83±0.38*
2.67±0. 21*θ
3
1.1
2.67±0.22*
3.17±0.19 θφ
4.0±0.36*
2.67±0. 21*θ
4
1.95
2.3±0.21* θ
3.0±0.18 θφ
3.0±0.37*
2.30±0.21*θ
5
0.98
3.3±0.22*
3.67±0.19*θ
3.5±0.54*
3.0±0.36*θ
6
1.10
3.0±0.37*
3.0±0.0* θφ
4.67±0.21*
2.33±0.21*θ
7
1.1
1.67±0.22
2.83±0.19 θφ
3.83±0.1*
1.67±0.
14
1.95
3.0±0.37* θ
3.67±0.28* θ
2.67±0.21*
2. 67±0.21*θ
16
1.45
2.0±0.0* θ
3.30±0.21* θφ
2.67±0.21*.
2.0±0.00*θ
19
1.45
2.67±0.22*
3.5±0.18* θ
4.0±0.63*
2.33±0.21*θ
20
1.95
2.67±0.22*
3.5±0.18* θ
3.67±0.76*
2.33±0.21*θ
21
0.98
2.0±0.0* θ
3.0±0.19 θφ
2.67±0.21*
2. 67±0.21*θ
22
1.45
2.3±0.42* θ
2.83±0.19 θφ
3.0±0.37*
2.33±0.21*θ
Analgesic effect
SCMC 0.5% sodium carboxymethyl cellulose ¥ P < 0.001 vs. non-arthritic control. Ɵ P < 0.01
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Analgesic effect
The analgesic activity was determined by pain tolerance and pain scoring assay. The results
(Table S 1) revealed that compounds 1, 4, 14, 16, 21 and 22 exhibited similar and higher
analgesic effect by pain tolerance assay. On the other hand the remaining tested compounds
have also shown good activities, all the tested compounds exhibited activities higher than the
reference drug in Pain scoring analgesic assay . By comparing the obtained results of antiinflammatory and analgesic activities of the compounds reported in this study to their structures,
the following structure activity relationship (SARs) were postulated: (i) Thiazole derivative 1 is
more potent than reference which may be attributable to presence of thiazole moiety. (ii) Most of
the other tested compounds exhibited activities higher than the reference and 1 which may be
due to the seleno moiety. (iii) Compound 4 and 7 were more potent than 3 and 5 which may be
due to the dimerization of 3 and 5 or the presence of the bisthiazole and diselenide moiety. (iv)
Compound 16 is more potent than 14 which may be due to the conversion of the cyanoacetamide
moiety into the coumarin moiety.(v) Compound 19 and 20 were more potent than 21 and 22
which may be due to the dimerization of 19 and 20 or presence of the bisthiazole,
bisnaphthoquinone and diselenide moiety (Figure S 1)
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N
H3C
HO
H
N
S
O
Ph
N
O
NH2
S
N
S
O
CH3
thiazol moiety increace activity
Meloxicam
1
Ph
N
Dimerization
Ph
Se
N
S
enhance the activity
N
H
R
Se
Se
Ph
NC
3, 5,19, 20
S
N
H
R
S
N
N
H
R
4, 7, 21, 22
3, 4, R=H
5, 7R=COCH3
O
19, 21,R=
HO
O
Ph
N
Se
NC
NH
S
O
20, 22,R=
O
O
Ph
conversion of cyanoacetamide
moiety into coumarin moiety Se
CN
increase the activity
NC
14
O
N
S
N
H
NH
O
16
Figure S 1: Structure activity relationship of the more potent antioxidant compounds
Material and Methods
Animals
90 Sprague-Dawley rats (200-250 g) were obtained from the animal house of Mansoura Faculty
of Medicine and fed on a standard rat chow and water ad libitum. Animal care and experiments
were performed in accordance with the guidelines established by the NIH Guide to the Care and
Use of Laboratory Animals, NIH publication no. 86-23. Rats were housed under similar standard
laboratory conditions. The animals were divided into two main groups (non-arthritic control,
arthritic group). In the arthritic group, all rats had been inoculated by the reagent of Collagen-
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Adjuvant arthritis into the left paw pad. Rats which developed right paw arthritic manifestations
after 45 days were divided into four groups as follow, arthritic control, piroxicam treated groups.
The compounds were given orally via gastric tube daily for seven days, 45 days after CollagenFreund’s injection. This is a modified form of the previous models.19 This model of Collagen–
Adjuvant Arthritis19-21 is considered to be a representative of rheumatoid arthritis or ankylosing
spondylitis in humans. Collagen II-Freund’s adjuvant emulsion (0.1 mL) was injected
intradermal into the left hind foot paw-pad of each rat (if no arthritis developed within 4 weeks,
some of the animals were challenged by a second inoculation). After 45 days, the systemic
arthritis developed in both hind paws.22 Most of the previous compounds are insoluble in water.
These were suspended in 0.5% sodium carboxymethyl cellulose [CMC]. The determined doses
were injected intraperitoneally. The doses are calculated according to reference .23
Measurement of joint inflammation
Rheumatoid index was based on severity and extent of the erythema and edema of the
periarticular tissue, and the enlargement, distortion or ankylosis of the joints. Its inflammation
was graded from 1 to 4 .24 Grading of 4 was when the joint was distorted and ankylosed, 3 when
markedly enlarged, 2 when erythematous with edema, and 1 when normal. 24 Each of the six nonarthritic, non-treated rats had a score of 1. Right paw pad thickness and joint scoring were
measured on the 7th day after starting drug treatment (45 days after complete Freund’s adjuvant
injection).
Edema thickness
The adjuvant induced inflammation in the contralateral paw pad (hind right paw pad) was
measured by the paw edemameter (paw thickness). It was carried out 45 days after complete
Freund's adjuvant injection .24,25
Pain tolerance measurement (Analgesic effect)
Pain tolerance measurement (Analgesic effect) by pain tolerance measurement right paw pad
pressure tolerance (Analgesimeter: Ugo Basile, Italy).
For assessment of the analgesic activity of the used drugs, pressure was applied by the
analgesimeter on the rat pad of the right paw. The pressure was increased gradually (a certain
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number of grams per second until the rat either squeaks or tries to withdraw its limb). The force
of pressure was continuously monitored by a pointer moving along a linear scale. Increased
pressure tolerance of drug treated rats indicates analgesic activity of the administered drug.
25
24,
This measurement of pressure tolerance was done at the 7th day of drug treatment (45 days after
complete Freund’s adjuvant injection).
Proximal joint (right ankle) mobilization tolerance (pain scoring)
This mobilization tolerance was graded from 1 to 4. Degree 1 corresponds to tolerance of
complete flexion 90o; degrees 2, 3 and 4 correspond to increasing degrees of maltolerance
according to a rat hind limb withdrawal, squeaking and when the flexion becomes painful.
Degree 4 corresponds to squeaking with just initiation of flexion. Each of the six non-arthritic,
non-treated rats had a score of 1.24 This measurement of mobilization tolerance was done at the
7th day of drug treatment (45 days after complete Freund’s adjuvant injection).
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Figure S 2: 1H-NMR for compound (3)
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Figure S 3: 13C NMR for compound (3)
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Figure S 4: 1H NMR for compound (5)
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Figure S 5: 13C NMR for compound (5)
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Figure S 6: 1H NMR for compound (7)
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Figure S 7:13C NMR for compound (7)