Chapter II
Synthesis of 1,2,4-triazole derivatives and their
Biological Activities
2o0. Introduction,
Triazole refers to either one of a pair of isomeric chemical compounds with molecular
formula C2H3N3, having a five membered ring of two carbon atoms and three nitrogen
atoms. The two isomers are: 1,2,3-triazole and 1,2,4-triazole.
Chapter II
-IMH ^ N H
1,2,3-triazole 1,2,4-triazole1 2
1,2,4-Triazole and its derivatives represent one of the most biologically active class of
compounds and are associated with diverse pharmacological activities such as
antibacterial, antiftingal, hypoglycemic, antihypertensive and analgesic properties
[1-5], The substituted 1,2,4-triazole nucleus is particularly common and can be found
in marketed drugs such as fluconazole, terconazole, rizatriptan, alperazolame and
triazolame [6].
2,1. Recent AdvaEcements
Recent literature survey demonstrates that the 1,2,4-triazoles are becoming of great
practical significance. 1,2 ,4-triazole derivatives possess analgesic, antipjnretic and
antiphlogistic properties. A group of 1,2,4-triazole-5-ones and their mannich bases
have shown antitubercular activity [7] and have been investigated with regard to their
mode of action. Few examples of biologically active 1,2,4-triazole derivatives are
given in table 1.
14
Table 1. Biologically active 1,2,4-triazole derivatives
Chapter I I
S.NO . lU P A C Nam e Structure B io logica l A ctivity
1
2 -{ (4-methylpiperazin-1 -
yl)methyl I- -4-phenyl-5-
(pyridin-4-yl)-2H -1,2,4-
triazole-3 (4H)-thione3
Antibacterial [ 8 ]
2
2'[5- {(1 -m ethyl-1 F I - p y i T o l -
2-yl)methyr}-4-phenyl-4H-
1,2,4-triazol-3-ylthio]-N-(4-
chlorophenyl)acetamide
“ 64
Antimicrobial [9]
3
7 -(4 -{(lH -I,2 ,4 -triazo l-l-
yl)methyl} benzyl oxy]-4-
methyl-2H-chromen-2-one
CH3
X X a
5
Antibacterial [10]
4
2-[5-{4-(4-brom ophenyl
sulfonyl)phenyl} -4-p-tolyl-
4H -l,2 ,4-triazol-3-yith io]-l-
phenylethanone9 °
H3C
6
Antifungal [11]
5
5-[{(E )-5-(4-
bromobenzylideneamino)-
4H -l,2,4-triazol-3-
yI}m ethyl]-4-(4-
fluorophenyl)-4H-1,2,4-
triazoIe-3-thiol
^ -F
Anticonvulsant [12]
15
Chapter I I
6
7'methoxy-5-phenyI-
[i,2,4]triazolo[4,3-
a]quinoline N
8
Antibacterial [13]
7
4-(4-Brom ophenyl)-5-(3-
clilorophenyl)-2,4'dihydro-
3 H -1,2,4 -triazole-3 - thi one
N—NH
c\ \VB r
9
Anti-inflammatory
[14].
8
3-(4-chlorophenyl)-5-
(ethylsulfonyl)- IH -1,2,4-
triazole
N—NH
CHj1 0
Anti-inflammatory
[15].
In view of the biological impovtance of these 1,2,4-triazoles, several methods have
been developed for the synthesis o f this class o f compounds. Some methods are listed
below,
2.2, Synthesis of 1 ,254-tHiazoIe derivatives.
2.2,1 Thiosemicarbazide (11) on cyclization in alkaline medium afford 1,2,4-triazoles
(12, 13) [16,17].
N -N
IR
11 12
Scheme-113
R = Phenyl, 4-Methoxy phenyl, 3-methyl phenyl, 2-Flouro phenyl
2.2,2 Thiosemicarbazides (14) on heating with triethylamine in ethanol cyclizes to
give 1,2,4-triazoles (15) [18].
16
Chapter II
o,R
N(C2Hg)3
14S chem e-2
R ^substituted Aryl
R
\ N -N H
15
2.23, Thiosemicarbazide on refluxing with 1,1 cyclopropane dicarboxylic acid (16)
and SOCI2 in alkaline medium cyclizes to give I,l-to(3~thio-],2,4-triazol-5-yl)
cyclopropane (17) [19].
HO'
O O i. SOCI2 , H2 NNNCSNH2
ii, NaOHOH
16
HS^ "n H
N -N N-^N'! W II ^
N H
'SH
Scheme-3 17
2,2.4. 2-Chloro-6-niethoxy-4-phenyl~quinoline (18) on refluxing with substituted
acylhydrazides in a nitrogen atmosphere yield 7-Methoxy-l"(4-methoxyphenyl)-5-
phenyl-1,2,4- triazolo [4,3-a] quinoline (19) [20].
i. RCONHNH2 .ii. n-butanol, 140-150 °C 20-40 h.
18Sciieme 4
'OCH3
2.2,5. (2£^-4-phenylquinolin-2(l/f)-one hydrazone (20) when refluxing with formic
acid in ethanol yield 5-phenyl-3,3a-dihydro [1, 2, 4]triazolo[4,3-a]quinoline (21) [21].
-I * * 2 I pormic acid N -^ ^ N ii. Ethanol
20 Scheme-5 21
2.2.6. l,2,4-Triazole-3,5-diamine derivatives (23, 24) were synthesized in moderate
to high yields in one-pot reaction from the corresponding isothiocyanates (2 2 ), mono-
17
Chapter II
substituted hydrazines, and sodium hydrogen cyanamide in the presence o f l-(3-
dimethylaminopropyI)-3 ethylcarbodiimide hydrochloride. Typically, two target
compounds were obtained, but high regioselectivity to one isomer was observed when
aromatic and sterically bulky hydrazines were used. Examples with a detailed
representative procedure are given below [22],
R iN C S
22
i. NaNHCNii, R2 NHNH2 ,
EDCN23
NH,
R2H
N NH2
24
Scheme- 6 R1/R2 = Alkyl, Aryl
2.2.7, Cyclo condensation reactions of phenyl selanyl propionate, using phenyl
selanyl group as the precursor of terminal double bond is critical to the success o f the
reactions [23].
O
PhSe-oH
25
R2-N=P-NH-R,O-dichlorobenzene
Scheme-7
R1/R 2 = Alkyl, Aryl
PhSe-
2.2.8. The N-(4-methylsulfonylphenyl)aryl carbohydrazonamides (27) undergoes a
ring closure using l,r-thiocarbonyldiimidazole and subsequent alkylation to afford 5-
(4-halophenyl)-4-(4-(methylsulfonyl)phenyl)-2H" 1,2,4-triazole-3(4H)-thione (28)
[24].X X
H3CO2S 1 , 1 '-thiocarbonylimidazote H3 CO 2 S
'T >>~-NH
27 Scheme- 8 28
2.2.9. The reaction of N, N-dimethylfomiamide azine (29) with primary amines
mediated by p-toluene sulfonic acid gives the triazole (31) as shown in scheme-9. The
18
driving force o f tliis reaction is the release of dimethyl amine as well as the stability o f
the triazole formed (31) [25].
H
N -N pTosOHyj3 M_// + H2 N~R ------------------- - I N~RMeaN—\ _2 HNMea
H29 30 31
R = Alkyl/Aryl
Scheme-9
1,354-oxadiazole on reaction with the primary amines gives 3,5-disubstituted 1,2,4-
triazole (34) [ 26],
RipTosOH
Chapter I I
p lO S U I - 1N
CH3 CH3
32 33 34
R1 /R2 = Aikyl/Aryl
Scheme-10
2.2.10. (Z)"4-chloro-N’-(ethoxy(p-tolyI)methylene)benzohydrazide (35) undergoes
cyclization in presence of hydrazine monohydrate to produce 4-amino-3-(p"
chlorophenyl)-5-p-tolyl”4H~l,2,4-triazole (36) [27].
ClpHa — / i. Propanol
O-CH2 f A ii. NH2NH2.H2O / = \
NH\ NH,
3 5 Scheme-11 36
2,2.11. Reaction o f resin bound S-methyl isothiourea with carboxylic acids yielded
resin-bound S-methyl-A^- acylisothiourea (37) which reacted with hydrazines under
mild conditions to afford the corresponding resin-bound 3-amino-1,2,4-triazoles (38) with regioselactivity [28].
19
Chapter II
H3Co s o
O N '
3 7
Ri
i. D M Fii. R 2 N H N H 2
iii.TFA /D C M
S chem e-12
H2N-N ~ N
38
■Ri
2.2.12. When substituted phenylthiourea (39) is refluxed with 2"(aryloxy)alkanoic
hydrazides (40) in pi'esence of pyridine, it gave 1,2,4-triazole (41). The completion
o f the reaction was assured by the ceasing o f the methyhiiercaptane evolution [29].
H-N^NHa
O,Pyridine, reflux ^ / / \ ___^ ^ 'N H
N-AoR
39 40
R= Me, Cl, Br etc
Scheme-13
41V
R
2.3. Reactions of 1,2,4-trlazoIes
2,3.1. Triazole (43) on condensation with heteroaromatic acids in presence of
phosphorus oxychloride produces a series o f triazolo thiadiazoles (42, 44, 45) [30].
C OOH
Of>„POCI3
R=Phenyl, Methyl,H
Ar
N'Nn " s h
NH2
43
R’
N"N
POCI3
C O O H N '- fv i
APOCI3
R= OH, Cl, F R'=OH, H
Scheme-14
20
R '" 'N R
44
2.3.2. 1,2,4-triazole-3”thiol (46) on reaction with ethyl bromoacetate in the presence
of sodium ethoxide or triethylamine, hydrazine hydrate, CS2 and KOH leads to the
fonnation of 5-[{(4- phenyl-5-pyi‘idin-4-yl-4H-l,2,4-triazol"3-yI)thio}methyl]-1,3,4-
oxadiazole-2“thiol (47) which on reaction with 2-(4-morpholino) ethylamine in the
presence of formaldehyde solution is converted to the coiresponding mannich base
derivatives (48) [31].
Chapter I I
/ = \ N"nh
N -N iii. CS2, KOH r 11 ii. Formaldehyde
.Abs.Alcohol, Na ''— ^ j O Sii.Ethylbromo acetae i- Ethanol \ __
47
/==\ 7 'V n- n/>— ^ '1
N, NH
46 N S 49
48
Scheme 15
2 A. Present W ork
The 1,2,4-triazole derivatives and their mannich bases represent an important class of
heterocyclic compounds which are known for their broad spectmm of biological
activities including antimicrobial, anti-inflammatory, analgesic and many other uses
[32-38]. As resistance to anti-inflammatory and antimicrobial drugs is widespread,
there is an increasing demand for the identification of novel structure leads that may
be used iii designing new potent and less toxic anti-inflammatory and antimicrobial
agents. Several 1,2,4-triazole based antimicrobial agents have been synthesized to
develop new molecular entities with high potential against various microbial strains.
Further several nonsteroidal anti-inflammatory drugs (NSAIDs) have been developed
so far to heal the inflammation. There are number o f NSAIDs present in the market to
heal the inflammation but majority of them cause adverse side effects like gastric
ulcer [39], kidney damage [40] and some o f the NSAIDs also cause hepatotoxicity
[41]. During the last few decades, a considerable attention has been devoted to the
synthesis of various 1,2,4-triazole derivatives [42-44]. Several examples of NSAIDs
having triazole moieties have been cited in the literature. Among them, 1,2,4-triazole
3-thiol derivatives are of particular interest and have been studied in recent years
[45-48],
21
Most o f the NSAIDs having biphenyl derivatives substituted with an aromatic or
heteroaromatic aryl nucleus show better anti-inflammatory activity. Some o f the
biaryl acid derivatives have been evaluated as potential anti intlammatory agents
through the inhibition of 14 kDa human platelet phospholipase A2 (HP-PLA2) [49].
Some of the biphenylic compounds that contain free carboxyl groups cause ulceration
[51-52]. Moreover quinoline moiety was also found in several bio-active molecules
(Quinine, Ciprofloxacin).
In view of the biological potentials of quinoline, biaryl and 1,2,4-triazole moieties as
anti-inflammatory and anti microbial agents, we considered it of interest to synthesize
some small molecular high affinity ligands by conjugating these bioactive
pharmacophores. The synthesis and biological activities of these conjugates are
presented in the following four sections.
Section 1. Biphenyl based 1,254-triazoIe derivatives and tlieir ijiologkal activities
Section 2. Biphenyl based hydrazones of 1,2,4-triazaIe deiivatives and their biological activities
Section 3. Synthesis of novel 3~mercapto 1,2,4-triazole derived mannich bases and their biological activities
Section 4. QwlnoMne based 1,2,4-triazole derivatives and their biological activities
Chapter II
22
Chapter II
Section 1
Synthesis of biphenyl based 1,2,4-triazole
derivatives and their Biological Activities
Chapter II, Section 1
2.4.1.0. Introdwctfon
As discussed in introductory part of this chapter, triazole derivatives exhibit different
biological activities. In view of the importance of 1,2,4-triazoles and biphenyl
systems, a focused library of 1,2,4-triazoie based conjugates have been synthesized
and evaluated for their anti-inflammatory, analgesic, ulcerogenic and antimicrobial
activities. The synthetic route is given below.
Schenie-I
50
OH
Dry acetone anhydrous K2CO 3
ethylchloroacetate reflux 1 0 hr
/)--- ^O-CHaCOOCaHs
51Abs. alcohol NHzNHs.HaO reflux, 3 hr.
52
Abs. alcohol Arylisothiocyanate reflux, 4-6 hr.
\\ /yOOH,COUHHH~^^53-59
Abs. alcohol TEA, reflux, 6 - 8 hr.
r 'i 'SHO
N"NH
60-66Rl J
Abs. alcohol Na Metalsubstituted phenacylbromide reflux 4-6 hr.
o-r i
'N 'S -R o
67-105R,
Reaction of /?-hydroxybiphenyI (50) with ethylchloroacetate in presence of potassium carbonate in anhydrous acetone afforded ethyl 2-(biphenyl-4-yloxy) acetate (51)
23
Chapter II, Section 1
which readily yielded 2-(biphenyl-4-yloxy) acetohydrazide (52) by refluxing with hydrazine monohydrate in absolute alcohol.Compound (52) was then converted into corresponding thiosemicarbazide (S3-59) by
reacting with different substituted aryl isothiocyanates in absolute alcohol.
The thiosemicarbazides (53-59) were cyclised into respective substituted S-mercapto-
1,2,4-triazoles (60-66) using tri ethyl amine in absolute alcohol. Finally S-alkylation of
3 -mercapto-1,2,4-triazoles with substituted phenacyl bromides and other alkyl halides
leads to the formation of target molecules (67-105). All the synthesized compounds
have been completely characterized on the basis o f their detailed spectral data and this
entire series of new compounds was evaluated for different biological activity. The
physical data o f all the final compounds are given in table 1 .1 .
Table 1.1. Physical data of biphenyl based 1 ,2 ,4-triazoIe derivatives.
Compounds Structures Yield (%) m.p. (°C )
67
6 ^
72 164-166
68 67 170-172
69
6 ^
70 132-134
70
6 “ =
74 138-140
71
6
74 138-140
72
6
68 168-170
73
6
74 174-176
74
°"=».
70 156-158
24
Chapter II, Section 1
75
GHj
79 153-155
76
° " c H3
82 150-152
77 78 157-159
78
° "C H 5
84 168-170
79
° '-C H j
72 173-175
80
° " C H i
78 144-146
81
F
72 160-162
82
F
74 156-158
83 Q _ < Q ^ 0 s
F
85 172-174
84
F
62 144-146
85 O O o - ^ l ' ^ X r ' ^0
F
68 168-172
25
Chapter II, Section 1
86
0F
68 142-144
87
F
69 140-142
88
Cl
68 154-156
89 NI'N 0
Cl
72 183-184
90
Cl
75 178-180
91
Cl
68 192-194
92 82 162-164
93 76 143-145
94 SO 164-166
95Q ^ O ^ C H ,
NOa
78 198-200
96
9HQz
76 158-160
97
NOj
66 168-170
26
Chapter II, Section 1
2,4,1.1. Results and discussion
2.4.1.1J. Analytical
A focused library of thirty-nine new compounds 67-105 were synthesized starting
from p-hydroxybipheny] as outlined in Schenne-I. O-alkylation o f /7-hydroxybiphenyl
with a-chloroethylacetate was confirmed by the 'H-NMR spectrum. The appearance
of two signals at 5 1.85 (t, J - 5.6 Hz, 3H) and 3.92 (q, J ~ 5.8 Hz, 2H) suggested the
presence of ethyl group and appearance o f a strong absorption at 1674 cm"' in IR
spectrum confimis the formation of ethyl 2-(biphenyl-4-yloxy) acetate (5 1 ) .
Compound 51 readily yielded 2-(biphenyl'-4-yloxy) acetohydrazide (52) by its
reaction with hydrazine monohydrate. The formation of this hydrazide was confirmed
by the presence of signals due to NH-NHa group at 5 3.2 (br, s, NHa, 2H) and 6.29 (s,
N-H, IH) and absence of signals due to ethyl group. Compound 52 was then
27
Chapter II. Section 1
converted into corresponding thiosemicarbazide (53-59) which were confuined by
the presence of extra signals in aromatic region due to phenyl ring o f isothiocyanate
along with signals at 5 8.81-9.70 (CSNH), 9.21-10.34 (CONH) and 8.02-9.20 ppm
(Ar-NH-) respectively. Cyclization of compounds 53-59 to 60-66 was confirmed
from the IR and NMR spectral data. Absorption bands in the region 2680-2690 (S-H,
stretching) and 1594-1607 cm"' (C==N of triazole ring stretching) and the presence of
signal as singlet corresponding to thiol proton (-SH) in the range o f S 8.71-13.61 ppni
in 'H NMR spectrum confirmed the formation of 3-mercapto-I,2,4-triazole. Finally
S-alkylation of 3-mercapto-l,2,4-triazoles with substituted phenacyl bromides/benzyl
bromide/alkyl halides leads to the formation o f target molecules 67-105, which were
confirmed from IR (absorption bands for C=0, 1655-1687 , phenacyl substituted),
'■ C NMR spectrum (peaks in the range of 5 190-192 ppm for C=0, Phenacyl
subtituted) and from 'H NMR spectmm (absence of S-H signal).
All the newly synthesized compounds 67-105 were further screened for their
biological activities viz. anti-inflammatory, analgesic, ulcerogenic and antimicrobial
activities. These novel triazole derivatives have shown varied therapeutic efficacy in
vitro and in vivo experiments.
2.4.1.1.2. Biological Activity
2 . 4 . I . I . 2 . I . Anti-inflammatory activityThe results of anti-inflammatory activity are summarized in table 1.2 and figure 1.1
Among thirty nine compounds from this series, seven compounds viz. 68, 74, 88 , 95,
96, 97 and 101 were found to be active and showed significant inhibition (79.84,
64.69, 59.69, 52.12, 52,12, 59.69, 59.69 % and 83.03, 72.52, 52.55, 60.06, 65.01,
57.50, 67.56 %) in paw oedema at 3h and 5h respectively when compared to the
standard drug Tbuprofen (78.93 and 82.58 % at 3h and Sh respectively). Compound
68 has shown better inhibition compared to the standard drug Ibuprofen. From the
biological data, the structure activity relationship (SAR) can be shown as follows.
1, Compounds having no substitution on the aromatic ring attached to the 1,2,4-
triazollyl ring and the electron withdrawing group on the phenacyl aromatic
ring are showing better activity.
28
Chapter II, Section I
2. Substitution of electron donating groups on triazoUyi attached aromatic ring
decreases the activity but is comparable with the standard.
3. Whereas when the same aromatic ring was substituted with electron
withdrawing groups the activity significantly decreases,
4. Compounds having bulkier substitutions like -Br and -NO2 groups on the
phenacyl aromatic ring are showing potential activity.
5. Whereas the simple benzylated 3-mercapto l,2,4-triazoles viz. 69, 70, 71, 72,
73, 80, 84, 865 87, 104 and 105 significantly diminishes the activity.
The data suggests that the carbonyl functionality o f the phenacyl group may be
playing an important role in the enzymatic interactions responsible for anti
inflammatory activity. All data were analyzed by one-way ANOVA test followed
by Dunnett’s test in carrageenan induced rat paw oedema model in rats.
Table 1 .2 . Anti-inflammatory activity of biphenyl based 1,2,4-triazole derivativesCompounds Change in paw oedema volume (tnL)
After drug treatment
Anti-lntlammatory activity
% Inhibition
3h 5h 3h 51i
67 0.300±0.036*‘* 0.283±0.040"'" 54,54 57.50
68 0.133±0.02r** 0.113±0.033**' 79.84 83.03
69 0.466*0.061''“ 0.466±0.033"'' 28.64 29,39
70 0.483±0.032"" 0.460±0.1I8"* 26.81 30,93
71 0.583±0.060“ 0.388±0.068* 11.66 42.49
72 0.360±0.033*‘ 0,316±0.047" 44.54 52,55 -
73 0.416±0,030’ 0.310*0.044’* 36.96 53,45
74 0.233±O.Q49**‘ 0.183±0.040*** 64.69 72.52
75 0.423±0.058** 0,4I3±0.057™ 35.90 37.98
76 0.533±0.042 0.483±0.118™ 19.24 27.24
77 0.330±0,049” * 0.366±0,055’ 50.00 45.04
29
Chapter II, Section 1
78 0.483±0.030"" 0,460*0.117'’' 26.81 30.93
1 79 0.333±0.033 0.300±0.057**’ 49.54 54.95
SO O.SSSiO.OSO?" 0.430±0.034* 41,96 35.43
81 0.300±0.05r” 0,366±0,076" 54.54 45.04
82 0.298±0.057**‘ 0.300±0.057” ’ 54.69 54.95
83 0,295^0.046” * 0.279:t0.036*” 55.30 58.10
84 0,345±0.036” 0.331 ±0.039*’ 47.72 50.30
85 0.350±0,042*’‘ 0.233±0.091*” 46.96 65.01
86 0.400:1:0.044’ 0,4 10±0.091 * 39.39 38.43
87 0.300±0,033’** 0.3l0±0.090” ’ 49,54 53,45
88 0.266±fl.07r** 0.316±0.090*** 59.69 52.55
89 0.566±0.049’''‘ 0,420±0.l25™ 14.24 32.83
90 0.341 ±0.056” * 0,338±0.068"* 48.33 49.24
91 0.319*0.067"" 0.321 ±0.075*” 51,66 51.36
92 0.3!6±0.060" 0,233±0.033” ‘ 52,12 65.01
93 0,350±0.042** 0.300±0,057” 46.96 55.45
94 0.350±0.042" 0.266±0.042*’* 46.96 ~~60.06
95 0.316±0.060*** 0.266±0.049*“ 52.12 60.06
96 0.316±0.047‘“ 0.233±0.033*‘* 52.12 65.01
97 0.266±0,042 0.300±0.036 59.69 57.50
98 0.5S3±0.0S5™ 0.600*0.089"^^ 11.66 09.00
99 0.266^0.061“ * 0,2SO±O.O42‘*‘ 59.69 62.46
100 0.300±0.057"‘ 0.316±0.060’” 54.54 52.55
101 0.266±0.055 *" 0.216±0.030*** 59,69 67.56
102 0.383±0.047” 0.350±0,042*** 41.96 47.44
30
Chapter II, Section 1
103 0.366±0.033** 0.350±0.042 *' 34.54 47.44
104 0.383±0.040 * 0.300±0.036 *’ 41.96 54.54
105 0.316±0.030**‘ 0.283±0.065*’ 52.12 57.50
Ibuprofen 0.130±0.033*” O.llOiO.On*** 78.89 80.10
Control 0.660±0.033 0.666±0.033 - -
Data analyzed by one way ANOVA followed by Dunnett’s 't' test (n=6), *p<0.05, p<0.01 &***p<0.001 significantly different from standard; ns, not significant.
Anti-inflammatory activity
"13 hr U 5 h r
co15Zc5?
67 68 69 70 71 71 7 i 74 75 76 77 78 79 80 81 82 8 i 84 85 86
Compounds
co
JZc
Anti-inflammatory activity
mihr U5hr
Figure 1.1. Anti-inflammatory activity of biphenyl based 1,2,4 triazole derivatives.
31
Chapter II, Section 1
2.4.1.1.2.2. Analsesic activity
The results o f analgesic activity are summarized in table 1.3 and figure 1.2. Among
tested compounds, compounds 99 and 101 showed comparable analgesic activity
(reaction time) at 30 min., 60 min. and 120 min. respectively with the standard drug
Ibuprofen.
Table 1.3. Analgesic activity o f biphenyl based 1,2,4-triazole derivatives.
Compounds Mean value of Tail Flick Latency (sec)±SEM
0 min. 30 min. 60 min. 120 min.
68 2.16±0.298 2.85±0.367 3.91 ±0.529* 2.51±0.323
74 1.88±0.324 3.23±0.370"* 3.60±0.341“ 2.56±0.227“
88 1.93±0.33 3.04±0.284“ 2.84±0.302 “ 2.85±0.372
96 I.98±0.34 2.81±0.364“ 2.86±0.302 “ 2.83±0.371 ■“
97 2.02±0.310 2.90±0.238 “ 3.22±0.282"* 2.81±0.230“
99 2.07±0.368 3.81±0.257* 3.96±0.377“ 3.08±0.499*
101 1.98±0.303 3.88±0.342* 3.98±0.240* 2.95±0.238 “
Control 2.03±0.330 1.94±0.236 2.02±0.192 2.85±0.237
Ibuprofen 1.98±0.307 3.71±0.388* 3.95±0.473* 2.76±0.212”
Data analyzed by one way ANOVA followed by Dunnett’s 't' test (n=6), *p<0.05, p<0.01 &***p<0.001 significantly different from standard; ns, not significant.
Analgesic activity
•'O min *30 min ■•60 mm " * 1 2 0 mm
Figure 1.2. Analgesic activity o f biphenyl based 1,2,4 triazole derivatives.
32
Chapter II, Section I
2.4.1.1.2.3. Ulcerogenic study
When compared with ibiiprofen, compounds 74 did not cause any gastric ulceration
and disruption o f gastric epithelial cells at the given oral doses. Hence gastric
tolerance to this compound was better than that o f Ibuprofen indicating that
carboxylic group present in the Ibuprofen is responsible for ulceration, Bhandari et al.
[52], Stomach wall of Ibuprofen treated group at low power (lOx) photomicrograph
showed damage of the mucosa and the siib mucosa. Stomach wall of the same section
at high power (40x) photomicrograph showed desquamated epithelial cells in the
lumen whereas in tested compounds 68 and 10 1 treated animals, surface epithelial
damage was significant, slightly submucosal damage was seen however there was
lesser damage in comparison to the Ibuprofen. Stomach wall of compound 74 treated
animal showed no damage to any layer. The results are shown in table 1.4 and figure
1.3.
Table 1.4. Haematoxylin and Eosin Immunohistochemical staining of gastric ulcers
after ulcer induction in rats
CompoundsSurface Epithelial
DamageSubniucosal
DamageDeep Mucosal
DamageMuscular
Layer Damage
68 ++ " -
74 - -
88 ++ ++ -
97 ++ •H- “ -
99 ++ - -
101 + ■h " : -
Control - - - -
Ibuprofen +-H- 4-++ ~
Indomethacin -H-H -H-+
+. +++ (increased ulceration), - (no ulceration)
33
Chapter 11, Section 1
68 (lOX) 68 (40X)L U M E N
'4 (lOX) "4 (40X)L U M E N
88 (lOX) 88 (40X)
C ontinued.,.
34
Chapter II, Section 1
-t*-
l U M E N
- f « ; j ' ■
I * ’ InfUmm. c*IU f
101 (lOX)
mL U M C H
^ t l r '4 * K > y ^ v ? i ^ r S
Ibiipi ofeii (lOX)
101 (40X)
I ^ L UMI N * 1
Ibupi ofeii (40X)
;=.fv.
L U M I N
foiiri ol (lOX) conti oi (40X)
Figure 1.3. Haematoxylin and Eosin immunohistochemical staining of gastric ulcers after ulcer
induction in rats.
35
Chapter II, Section 1
2.4.1.1,2.4. Antimicrobial Activity
All the newly synthesized compounds were tested against various microbial strains
for their antimicrobial activity. The results are summarized in table 1,5, Some o f the
compounds showed moderate to good activity against certain strains. Compound 96
exhibited moderate activity against all the tested fungal strains. Compounds 71, 75,
79, 80, 93, and 94 were active against A. flavus. Compounds 75, 79, 92 and 102
exhibited moderate activity against A. niger. Compounds 96, 98 and 102 sliowed
moderate activity against C, albicans whereas compounds 70, 74, 75, 78, 80, 96 and
102 were active against C. krusie.
Compound 67, 78, 80 and 95 exhibited moderate activity against bacterial strains K.
pneumoniae. Compound 6 8 , 90, 94 and 97 were active against P. aeruginosa and E.
coli.
Table 1.5. Antimicrobial activity of biphenyl based 1,2,4-triazole derivatives.Compounds Atttifungal Activity (zone of Inltibition in mm) Antibacterial Activity (zone of inhibition in mm)
A . f l a v u s
200(100)Hg/disc
A . n i g c r
200(100)Hg/clisc
C .a lh i c a n s
200(100)Hg/disc
C .k r u s e i
200 (100) (.ig/disc
K .p n e u m o n i a e
200(100)Hg/diso
P .a e r u g i i t o s a
200(100)Hg/disc
R c o l l
200 (100) |xg/disc
S , a u r e u s
200(100) Hg/disc
67 18±1,5(I0±0,8)
6±0.5(-) 8±0.f)(6±0,4)
8±0.8(6±0,6)
14±!.2(6±0.6)
12±!.2(8±0.6)
8±0.7(6±0,6)
68 18±1.3(I0±I,2)
lOdbl(6±0.5)
8±0.6(-)
8±0.6(-)
8±0,5(6±0.4)
14*1.7 (10±1)
10±1.2(6±0,6)
8±0.7C-)
69 I4±l,6CI0±0,9)
8±0,6(-)
6±0,5(-)
9±1.2(fiiO.g)
I0±1.2(6±0.6)
8±0.6(-)■
■"
70 I0±0,5(6±0,3)
10±0.5(6)
6±0.4(-)
i0±0.6(8±0.4)
8±0.4(')
8±0.4 (-) 6(-) 8±0.4(6)
7J 18±l,6(8±0,<i)
10±l.2(6±0.4)
8±0,9(6±0,5)
8±0,9(6±0,6)
12±1,6(8i0.6)
12±1.2(8±0.7>
10±0.9(6±0,6)
8±0.6(-}
72 *■■ ' ■ ■ ■
73 **‘ ■ ■ ■ ■ ' ■
74 10±0,5(6±0,3)
I2±0.5(6)
6±0.4{-)
12±0,6(8±0.4)
8±0,4
(-)7±0,4 (-) 6 (-) 8
(6)
75 22±t.9(I0±1.2)
I4±0.fi(7±0.5)
lOil.l (6 ±0,4)
!0±0,8(fi±0.4)
I0±0,8(6±0.4)
8±1(-) ■
76 " ' ■’ ,■ ■
77 18±I,2(8±l,6)
I0±0.9(6±0.4)
10±0,9(6±0,4)
8±0.9(6±fl.4)
12±l,l(6±0,6) ■
10±I.2(-)
36
Chapter II, Section I
78 18±1 .«
( l O i l . l )
12±1 .2
(6 ± 0 ,4 )
8±0.8
(6 ± 0 .5 )
10±O .4
(6 ± 0 .5 )
1 4 ± 1 .6
(1 0 ± 1 .2 )
1 1 ± 0 .9
(-)
1 0 = tl.2
(S ± 0 .4 )
-
79 2 4 ± 1 .4
( I S )
!4 ± 1 .2
(10 )
)0 ± 0 .3 (8 ± 0 .
4)
6 ± 0 .3
(4 ± 0 ,2 )
U )± 0 .6
(6 ± 0 .3 )
1 0 ± 0 .6
(6 ± 0 .3 >
- 6
(-)80 2 2 ± I ,6
(1 6 i:S .2 )
I 2 ± M
(8)
lO iI .O
(8 )
lO iI .O
(8 )
14
(1 0 )
14
( iO )
“
81
'' “ -
82 I 8 ± l ,8 (
1 0 ± 1 .2 )
I2 ± 0 .9
(6±Q .4)
G±0.6
(-)
IO iO .6
(C>±0.4)
1 2 ± l .2
(8 ± 0 .7 )
1 2 ± l ,2
(9 ± 0 .6 ) (-)
8 ± 1 ,2
(6±0,6)
83 1 4 ± l,2
(8 ± 0 ,7 )
I 2 ± l . l
(8 ± 0 ,4 )
8± 0 .8
<6±Q,3)
8 ± 1 .2
(6 ± 0 .4 )
8 ± 0 .6
(6 ± 0 .3 )
I 4 ± l .2
(lOiO.O)
8±0,f)
(-)
6±0,6(-)
84
■ ■~ “ " "
85 8 ± 0 ,6
(-)
10 ± 0 .6
(6 ± 0 .4 )
8±fl,7
(-)
8 ± 0 .8
(6 ± 0 .4 )
8 ± 0 .6
(6 ± 0 .4 )
10±0 .8
(6±0.6)
6 ± 0 ,3
(-)
6 ± 0 ,3
(-)
86 M i l . 2
(6 ± 0 .3 )
10± 1 ,2
(6±0.6)
8±(),6
(5 ± 0 ,3 )
9 ± 0 .8
(7 ± 0 .6 )
I 2 ± l .2
(8 ± 0 .6 )
** ■ "
87
■ '■ ~ •* “
88
■" “ “ , *•
8 9 12±1 .6
(fi± 0 .4 )
10±1,4
(6±0,3)
8--bO,3
(6 ± 0 ,4 )
fi± 0 .4
C-)
8 ± 0 .5
(-)
1 2 i l . 3
(6)
1 0 ± 1 .)
(8±0.S)
8 ± 0 .4
(-)
90 I8 ± l .3
(1 0 ± 1 ,2 )
I0±1
(6 ± 0 ,5 )
8 ± 0 .6
(-)
8 ± 0 ,6
(-)
8 ± 0 .5
(6± 0 .4>
1 4 ± ! .7
(IO±l)
I0±1.3
(6±().6)
8 ± 0 .7
(-)
91 I2 ± l .4
(6 ± .4 )
I0 ± l ,5
(6 ± ,5 )
8 ± l , f i
(6 ± .6 )
“■
8 ± ,8 {-) 8± .5
(-)
*•
92 I9 ± I ,6
(8 ± .6 )
I 4 ± l .4
(2 ± ,7 )
8 ± .5
6 ± .3 )
6± .3
(-)
6± .3
(- ) ■
I 0 ± L 2
(6 ± ,5 )
9 ± ,8
(-)
~
93 2 4 ± 1 ,8
(1 2 ± I .2 )
8± 0 .8
(-)
8 ± 0 .6
(6 ± 0 .3 )
8 ± 0 ,4
(6 ± 0 .4 )
10± 0 .6
(6 ± 0 .3 )
6 ± 0 .6
f4 ± 0 .3 )
“ "
94 2 2 ± 1 .6
(1 4 ± 0 ,S )
I2 ± 0 ,6
(8 ± 0 .4 )
8 ± 0 ,4
(6 ± 0 .3 )
f)±0.3
(-)
I 2 ± l .2
(8 ± 0 .6 )
1 4 ± 1 .3
(10±0.6)
10a=1,0
(6 ± 0 .3 )
■
05 lO iI.J
(8 ± 1 .0 )
I2 ± l .6
(10=t0.0)
8 ± 0 .9
(-)
8±I.O
(fi±Q.6)I 9 ± l .6
(! f l± 0 .4 )
9±0.(i f-) I 0 ± ! .2
(')
■*
96 2 2 ± 1 .8
( I 2 ± r .6 )
I8 ± ,8
(1 0 ± ,4 )
1 2 ± 1 .I
(8 ± .5 )
10±.S
(8 ± ,6 )
1 0 ± 1 .2
(6 ± ,4 )
6 ± .5
(-)
■*
97 12± 1 .2
(SiO.O)
I 2 ± I .
(S±0.6)
1 4 ± 1 ,2
(lOiO.6)
10±1,0
(6 ± 0 .3 ) ■
98 10± 0 .6
C 6±0.5)
12 ± 0 ,9
(6 ± 0 ,4 )
1 2 ± 1 .2
(8 ± 0 .4 )
11± 0 .8
(8 ± 0 .6 )
NT NT S ± 0 ,4
(-)
6 ± 0 ,3
(-)
99
‘" ■* “ - *■ *■
100 ** “ ” "■ ■ ■
101 ■ - - - - - - - "
37
Chapter II, Section 1
102 16±l.2(9:1:0.6)
14±0,9
(6±().r>)n±o,8(9=fc0.f,)
NT NT 8±0.6 (-) 6±0.3
(-)103
104 NT NT NT NT NT NT NT NT
105 NT N T 8±<).4
(-)6±0.3
(-)Fluconazole 22±1.8
([2±.9)
I6±l
(I0±.6)I4±1.2
{ 10±.)
I0±I,2
(8±.7)
NT NT NT NT
Amoxicillin NT NT NT NT 16±1
(1l2±.S)
I4±.8
nt»±.5>
12±.7(8±.4)
12±.f>(6±.3.)
C ontrol
NT: not tested; - : no zone o f inhibition
2.5.1.2. Experimental
2,5.1.2.1. Chemistry
All chemicals (reagent grade) used were commercially available. Melting points were
measured on a VEEGO-VMP-DS melting point apparatus and are uncorrected. ’H
NMR was recorded on a Bruker DPX 400, 300 instraments in CDClj/DMSO-dfi using
TMS as internal standard. 'H NMR chemical shifts (5) and coupling constants {J) are
given in ppm and Hz respectively. Mass spectra were recorded on either a Jeol JMS-D
300 instrument fitted with a JMS 2000 data system and operated at 70 eV or Maldi-
MS (AB-4800). Mass-spectrometric (MS) data are reported in m/z. Elemental analysis
was carried out using Elementar Vario EL III elemental analyzer. Elemental analysis
data is reported in % standard.
General procedure fo r synthesis o f Ethyl-2-bipheny-4-yloxy-acetate (5 1 )
To a mixture of ;?-hydroxybiphenyl (10 mmol) and ethylchloroacetate (10 mmol) in
50 mL anhydrous acetone was added 15g K2CO3. The suspension was refluxed for 20
h. After completion of reaction, the reaction mixture was filtered in hot condition, the
filtrate concentrated under reduced pressure and finally the crude product crystallized
from methanol in cold condition in 75% yield as white flakes; m.p, 40-42 ”C, Rf =
0.64 (n-hexane : ethyl acetate; 6:4).
38
Chapter II, Section 1
-o
pHgP “ CH2
o
IR (KBr, cm"’)
‘H NMR (300 MHz, CDCI3)
FAB-MS im/z)
Elemental Analysis
Calculated
Found
: 3055, 2921, 1674, 1108.
: 5 1.85 (t, J - 5.6 Hz, 3H), 3.92 (q, J = 5.8 Hz,
2H), 5.29 (s, O-CH2, 2H), 7.02 (d, J = 8.6 Hz,
2H), 7.24-7.31 (m, IH), 7.39 (d, J = 7.6 Hz,
2H), 7.53-7.54 (m, 4 H, Ar-H).
: 256 (M^), 257 (M^+1).
: Calculated for molecular formula C 16H 16O3:
: C, 74.98; H, 6.2.
; C, 74.68; H, 6.21%.
General procedure fo r synthesis <?/'2-{4-plienyIpheiioxy)acetohyclrazide (52)
The ethanolio solution of (51) (10 mmol) and hydrazine monohydrate (10 mmol) was
refluxed for 4-6 h. After the completion of reaction, the reaction mixture was cooled,
the white flakes so obtained was filtered and recrystallized from alcohol. Yield; 90%;
White flakes; ni.p.165-167 "C, Rf = 0.23 (/^-hexane: ethyl acetate; 4:6).
AO
\\ / - A
IR (KBr) cm'
NM R (300 MHz, CDCI3)
FAB-MS (m/z)
Elemental Analysis
Calculated
Found
: 3350, 3055, 2921, 1674, 1108.
: 5 3.20 (s, NH2,2H), 5.21 (s, 2H, O-CH2), 6.29 (s,
NH, IH), 7.51- 7.52 (m, Ar-H, 4 H), 7.40 (d ,./=
7.8 Hz, 2H), 7.03 ( d , 8.5 Hz, 2H), 7.25-7.32
(m, IH, Ar-H).
: 242 (M""), 243 (M'^+l).
; Calculated for molecular formula C 14H 14N 2O2
: C, 69.41; H, 5.82; N, 11.56
:C , 69.13; H, 5.84; N, 11.51%,
39
Chapter II, Section I
General procedure fo r synthesis o f thiosemicarhazides (53-59)
To a solution of hydrazide (52) (1 mmol) in absolute alcohol (50 ml) was added
ar yl i soth i o cyan ate (1 mmol) and the reaction mixture was refluxed for 4-6 h. After the
completion of reaction monitored by TLC, the reaction mixture was cooled to room
temperature and the white crystals so formed were filtered and crystallized from alcohol to yield the pure thiosemicarbazide (53-59).
l-[2~(4~Biphenyloxy)acetyl]~4-phenylthiosemicarbazide (53) Yield: 77%; White
crystals, m.p.138-140 "C, Rf = 0.25 (n-hexane ; ethyl acetate; 4:6).
.'1IR (KBr) cm'
‘H NMR (300 MHz, DMSO-tlfi)
FAB-MS (w/z)
Elernental Analysis Calculated
Found
; 3324, 2945, 1701, 1212, 1109.
: 5 4.68 (s, 2H, O-CH2), 7.03 (d, J = 8.6 Hz, 2H),
7.25-7.42 (m, 8H, Ar-H), 7.44-7.54 (m, 4H, Ar-
H), 8.92 (brs, IH, ArNH), 9.21 (brs, IH,
CSNH), 10.14 (brs, IH, CONH).
377 (M*'), 378 (M'^+1).
Calculated for molecular formula C21H 19N3O2S.
C, 66.82; H, 5.07; N, 11.13.
C, 66.48; H, 5.15; N, 11.25%.
l-[2-(4~Biphenyloxy)acetyl]-4(4-methoxyphenyl)thiosemicarbazide (54) Yield: 72%;
White crystals, m .p.l 76-178 "C, Rf == 0.24 («-hexane ; ethyl acetate; 4:6),O
.0
IR (KBr) cm"' : 3246, 2953, 1702, 1205, 1109.
'H NMR (300 MHz, DMSO-dfi) : 5 3.88 (s, 3H, O-CH3), 4.68 (s, 2H, 0 -CH2),7 .0 1 -
7.08 (m, 4H, Ar-H), 7.33 (t, / = 7.5 Hz, IH),
7.46 (t, J = 5.7 Hz, 2H), 7.61-7.47 (m, 6H, Ar-
40
Chapter II, Section I
FAB-MS (m/z)
Elemental Analysis
Calculated
Found
H), 8.92 (brs, IH, ArNH), 9.21(brs, lH,CONH),
9.12 (brs, IH, CSNH).
407 (M'"), 408 (M"'+l).
Calculated for molecular formula C22H21N3O3S.
C, 64.85; H, 5.19; N, 10.31.
C, 65.08; H, 5.22; N, 10.26%
]~[2-(4-Biphenyloxy)acetyl]-4(4-fluorophenyl)thiosemicarbazide (55) Yield: 76%;
White crystals, m.p.186-188 ”C, Rf = 0.20 (n-hexane; ethyl acetate; 4:6).
IR (KBr) cm"*
NMR (300 MHz, DMSO-de)
FAB-MS im/7)
Elemental Analysis
Calculated
Found
: 3244, 3070, 2950, 1683, 1202, 1106.
: 5 4.66 (s, 2H, O-CH2), 7.19-7.31 (m, 3H, Ar-H),
7,37-7.42 (m, 4H, Ar-H), 7.46-7.51 (m, 4H, Ar-
H), 7.88 (d, J = 6.4 Hz, 2H), 8.67(brs, IH,
ArNH), 8.96 (brs, IH, CSNH), 9.56 (brs, IH,
CONH).
:395 (M '),396 (M^+1).
: Calculated for molecular formula
C2iH,sFN302S.
: C, 63.78; H, 4.59; N, 10.63.
: C, 63.98; H, 4.53; N, 10.58%.
l-[2-(4-Biphenyloxy)acetyl]~4(4-chlorophenyl)thiosemicarbazide (56) Yield; 74%;
White crystals, m .p,183-185 ”C, R f - 0.23 (n-hexane; ethyl acetate; 4:6).
Cl
IR (KBr) cm -1 3322, 2958,1714,1195,1108.
41
Chapter n , Section 1
‘H NMR (300 MHz, DMSO-dfi)
FAB-MS (m/z)
Elemental Analysis
Calculated
Found
: 5 4.69 (s, 2H, O-CH2). 6.94 (d, J - 6.4 Hz, 2 H),
7.30 (t, J = 5.7 Hz, IH), 7.36-7.43 (m, 4H, Ar-
H), 7.47-7.52 (m, 6H, Ar-H), 9.21 (brs, IH, Ar~
NH), 9.70 (brs, IH, CSNH), 10.34 (brs, IH,
CONH).
: 411 (M^), 413(M"‘+2).
: Calculated for molecular formula
C2|H ,8C1N302S.
: C, 61.23; H, 4.40; N, 10.20
: C, 61.41; H, 4.48; N, 10.18%.
l~[2~(4-Biphenyloxy)acetylJ~4(3-chlorophenyl)thiosemicarbazide (57) Yield: 72%;
White crystals, m.p. 176-178 ”C, Rf = 0.20 («-hexane : ethyl acetate; 4:6).
Cl
IR (KBr) cm"
'H NMR (300 MHz, DMSO-de)
FAB-MS (m/35)
Elemental Analysis
Calculated
Found
: 3322, 2958, 1712, 1198, 1108.
; 5 4.68 (s, 2H, O-CH2), 6.94 (d, J = 6.4 Hz, 2H),
7.30-7.43 (m, 5H, Ar-H), 7.48-7.50 (ra, 6 H, Ar-
H), 9.20 (brs, IH, ArNH), 9.72 (brs, IH, CSNH)
10.34 (brs, IH, CONH).
: 411 (M^), 413 (M'"+2).
: Calculated for molecular fonnula
C2iH,8CIN30 2S.
; C, 61.23; H, 4.40; N, 10.20 ; C, 61.41; H, 4.48; N, 10.18%.
42
Chapter II, Section 1
l~[2-(4-Biphenylaxy)acetyl]~4(4~mtrophenyl)thiosemicarbazide (58) Yield; 69%;
Light yellow crystals, ra.p,198-200 ”C, R f - 0.31 (o-hexane ; ethyl acetate; 3:7),
IR (KBr) cm"*
NMR (300 MHz, DMSO-df,)
FAB-MS {m/z )
Elemental Analysis
Calculated
Found
H HNP
: 3246, 2958, 1693, 1208, 1113.
: 5 4.69 (s, 2H, O-CH2), 6.94 (d, J - 6.6 Hz, 2H),
7.26-7.52 (m, 7H, Ar-H), 7.94 (d, J = 8.6 Hz,
2H), 8.02 (brs, IH, ArNH), 8.38 (d, / == 8.6 Hz,
2H), 8.81 (brs, IH, CSNH), 9.58 (brs, IH,
CONH).
: 422 ( M^), 423 (M'"+l).
: Calculated for molecular formula C21H 1SN4O4S.
C, 59.70; H, 4.29; N, 13.26
: C, 59.92; H, 4.32; N, 13.20%.
l-l2-(4-Biphenyloxy)acetyl]-4(2~methylphenyl)thiosemicarbazide (59) Yield: 71%;
White crystals, m.p.138-140 "C, Rf == 0.26 (n-hexane : ethyl acetate; 4:6),
HaC
IR (KBr) cm'* t 3320, 2948, 1704, 1215, 1254, 1204, 1102.
'H NMR (20 0 MHz, DMSO-de) ; 5 2,38 (s, 3H, CH3), 4.67 (s, 2H, O-CH2), 6.86
(d, / = 6.4 Hz, 2H, Ar-H), 7.30-7.56 (m, IIH ,
Ar-H), 8.92 (brs, IH, ArNH), 9.21 (brs, 1H,CSN
H), 10.12 (brs, IH, CONH),
: 391 (M'^), 392 (M*'+l).
; Calculated for molecular fomiula C22H21N 3O2S.
; C, 67.50; H, 5.07; N, 10.73.
: C, 67,85; H, 5.11; N, 10.68%.
Maldl-MS (w /4
Elemental Analysis
Calculated
Found
43
Chapter II, Section 1
General method fa r synthesis o f S-[(hiphenyl~4~yloxy}methyl]4~aryl-3~mercapto-
(4H)-l,2,4-triazole (60-66)
To a solution of thiosemicarbazide (53-59) (5 mmol) in absolute alcohol (50 mL) was
added triethyl amine (1 mL) and the reaction mixture refluxed for 4-8 h. After
completion of reaction monitored by TLC, the reaction mixture was cooled to room
temperature and concentrated under reduced pressure (25 mL), the concentrated
solution was poured on crushed ice, the precipitate obtained was filtered off and
washed with cold water, dried and crystallized in ethanol to yield the pure
3-mercapto-l,2,4~triazoles (60-66).
S~[(Biphenyl-4-yloxy)inethylJ4~(phenyl)-3-m.ercapta-(4H)-l,2.4~triazoie (60) Yield: 78%; White crystals, m.p. 234-236 "C, Rf = 0.64 («-hexane : ethyl acetate; 4:6).
m (ICBr) cm”
‘ H NMR (300 MHz, CDCla)
Maldi-MS (m/z)
Elemental Analysts
Calculated
Found
Q N- N/y
N '^ S H
2912, 2690, 1607, 1110.
8 4.69 (s, 2H, O-CH2), 7.04 (d, J = 8.7 Hz, 2H,
Ar-H), 7.25-7.44 (m, 8H, Ar-H), 7.52-7.57 (m,
4H, Ar-H), 9.57 (brs, IH, S-H).
359 (M‘"), 360 (M'^+1).
Calculated for molecular formula C21H 17N3OS.
C, 70.17; H, 4.77; N, 11.69.
C, 70.09; H, 4.64; N, 11.58%.
5-[(Biphenyl-4-yloxy)methylJ4-(4~methoxyphenyl)-3~mercapto~(4H)-l,2o4-triazole
(61) Yield: 68%; White crystals, ni.pJ68-170 ”C, Rf = 0.63 (i^z-hexane : ethyl acetate;
4:6).
44
Chapter II, Section 1
• IR (KBr) cm-*
*H NMR (300 MHz, CDCI3)
Maldl-MS (m/z)
Elemental Analysis
Calculated
Found
: 3058, 2688, 1603, 1107.
8 3.88 (s, 3H, O-CH3), 4.93 (s, 2H, O-CH2), 6.91
(d, / = 6.6 Hz, 2H, Ar-H), 7.01 (d, J = 6.6 Hz,
2H, Ar-H), 7.30-7.36 (m, 3H, Ar-H), 7.42 (d, J
= 5.7 Hz, 2H, Ar-H), 7.49 (d, J - 6.9 Hz, 2H,
Ar-H), 7.52 (d, 5.4 Hz, 2H, Ar-H), 8.71 (brs,
IH, SH).
389 (M^), 390 (M’"-fl).
Calculated for molecular foiTnuia C22H 19N3O2S.
C, 67.84; H, 4,92; N, 10.79
C, 67.76; H, 4.89; N, 10.75%.
5-f(Biphenyl~4-yh)xy)methyl]4~(4-Jluorophenyl)"3-mercapto-(4H)~l,2.4"triazole (62) Yield; 76%; White crystals; m.p.186-188 ”C, Rf = 0.66 («~hexane : ethyl acetate;
4:6).
IR (KBr) cm-'
‘H NMR (300 MHz, CDCI3)
Maldi-MS (m/i:)
Elemental Analysis
Calculated
Found
// O\__//N- N
\ J i N ^ S H
y /F
3049,2929,2680,1603,1514,1105.
6 4.73 (s, 2 H, O-CH2), 6.94 (d, J = 6.3 Hz, 2H,
Ar-H), 7.21 (t, J = 8.3 Hz, 2H, Ar-H), 7.28 (t, J
= 5.4 Hz, IH, Ar-H), 7.37-7.41 (m, 4H, Ar-H),
7.51-7.46 (m, 4H, Ar-H), 8.73 (brs, IH, SH).
377 (M''), 378 (M^+1).
Calculated for molecular formula C21H 16FN3OS.
C, 66.83; H, 4.27; N, 11.13
C, 66.92; H, 4.25; N, 11.06%.
45
Chapter II, Section 1
5~f(Biphenyl-4-yloxy)methylj4-(4~chlorophenyl)~3-mercapto-(4H)-l,2,4-triazole
(63) Yield; 85%; White crystals, ni.p. 228-230 "C, Rf = 0.67 (^-hexane : ethyl acetate; 4:6).
IR (KBr) cm-'
‘H NMR (300 MHz, CDCI3)
MaMi-MS im/z)
Elemental Analysis
Calculated
Found
;3056, 2922, 2690, 1606.
: 5 4.52 (s, 2H, O-CH2), 7.03-7.06 (d, 8.7 Hz,
2H, Ar-H), 7.29-7.32 (t, 8,1 Hz, IH, Ar-H),
7.39-7.44 (m, 6H, Ar-H), 7.52-7.54 (d, 6.9
Hz, 2H, Ar-H), 7.54-7.57 (t, J - 8.4 Hz, 2H, Ar-
H), 8.71 (brs, 1H,SH).
: 393 (M'^), 394 (M'"-H).
: Calculated for molecular formula CaiHiGClNsOS.
; C, 64.03; PI, 4.09; N, 10.67.
: C, 64,08; H, 4.12; N, 10.63%.
S-[(Biphenyl~4-yloxy)methylJ4-(3-chlorophenyl)-3~mercapto-(4H)~l,2,4-triazole
(64) Yield: 78%; White crystals, m.p. 220-222 ”C, Rf = 0.68 (/7-hexane : ethyl
acetate; 4:6).
IR (KBr) cm‘
’H NMR (300 MHz, CDCI3)
Maldl-MS im/z)
ElemeBtal Analysis
; 3056, 2922, 2690, 1606.
: 5 4.74 (s, 2H, O-CH2), 6.89 (d, 8.4 Hz, 2H,
Ar-H), 7.25 (s, IH, Ar-H), 7.29-7.43 (m, 4H,
Ar-H), 7.47-7,52 (m, 6H, Ar-H).
: 393 (M^), 394 (M-'+l).
; Calculated for molecular formula C21H 16CIN3OS.
46
Chapter II, Section 1
Calculated
Found
C, 64.03; H, 4.09; N, 10.67.
; C, 64,09; H, 4.10; N, 10.63%.
5~f(Biphenyi~4-yloxy)methylJ4~(4~nitrophenyi)~3~mercapta-(4H)-l,2.4-triazole (65)
Yield: 89%; Light yellow crystals, ni.p. 221-222 ”C, Rf = 0.64 (n-hexane : ethyl
acetate; 4:6).
-1m (KBr) cn f
‘H NMR (300 MH*, CDCI3)
Maldl-MS (m/z)
Elemental AHalysIs
Calculated
Found
O2N
2912, 2690, 1604, 1102.
6 4.81 (s, 2H, O-CH2), 6,96 (d, J = 6.6 Hz, 2H,
Ar-H), 7.28 (t, J = 6.9 Hz, IH, Af-H), 7.32-7.52
(in, 6H, Ar-H), 7.65 (d, J = 8.8 Hz, 2H, Ar-H)
8.39 (d, 8.6 Hz, 2H, Ar-H), 8.93(brs, IH, S-
H).
393 (M"), 394 (M^+1).
Calculated for molecular formula C21H 16N4O3S.
C, 62.60; H, 3.99; N, 13.85.
C, 62.69; H, 4.01; N, 12.79%.
5-[(Biphenyl~4-yloxy)niethyl]4-(2-methylphenyl)~3~mercapto-(4H)-l,2.4-triazole
(66) Yield: 72%; White crystals, m.p. 186-188 °C, Rf = 0.68 («-hexane : ethyl
acetate; 4:6).
IR (KBr) cm"
‘H NMR (300 MHz, CDCI3)
: 2934, 2686, 1604, 1594, 1108,
: 5 2.54 (3H, s, GH3), 4.68 (s, 2H, O-CH2), 6.89
(2H, d, J = 6 .6Hz, Ar-H), 7.43 (m, 7H, Ar-H),
7.51-7.54 (m, 4H, Ar-H), 8.31 (brs, IH, S-H).
47
Chapter II, Section I
MaIdi“MS (m/z) Elemental Analysis
Calculated
Found
373 (M"), 374 (M'"+l).Calculated for molecular formula C22H|qN.')OS.
C 70.75; H 5.13; N 11.25
C 70.82; H 5.10; N 11.30%.
General method for synthesis o f l(Aryl)~2-[S~{(biphenyl-4~yloxy)methyl}4-aryl~3-
mercapto~(4H)~l,2.4~tnazol-3~ylthw)J ethanone/ethane (67-105)To a solution o f 3-mercapto-l ,2,4-triazoie (62-66, 0.5 mmole) in absolute ethanol (50
ml) was added sodium metal (0.5 mmole) and different substituted phenacyl bromide
(0.5 mmole) and the reaction mixture was refluxed for 1-6 h. After completion of
reaction monitored by TLC, the reaction mixture was concentrated under reduced
pressure (25 ml), the concentrated solution cooled and the crystals obtained were
filtered, washed with water, dried and recrystallized firom dichloromethane (DCM)
and methanol to yield the pure compound (67-105).
2-{S-[(4-Biphenyloxy)methylJ~4-(phenyl)-4H-l,2,4-triazol-3-ylthio}~l-(4-hromophenyl) ethanone (67) Yield; 72%; White crystals, m,p: 164-166 "C, Rf = 0.43
(«-hexane : ethyl acetate; 4:6).
IR (K B r)cm "‘ : 2907, 1686, 1570, 1093.
'H NMR (400 MHz, CDCI3) : S 4.97 (s, 2H, S-CH2), 5.02 (s, 2H, O-CH2), 6.87
(d, J = 6.3 Hz, 2H, Ar-H), 7.23 (t, J - 5,4 Hz, IH, At-H), 7.34-7.41 (m, 1IH, Ar-H), 7.56 (d, J
= 6.3 Hz, 2H, Ar-H), 7.83 (d, J = 6.0 Hz, 2H,
Ar-H).
NMR (100 MHz, CDCI3) : 6 41.01, 63.35, 115.18, 126.79, 126.81, 126.91,
126.95, 128,26, 128.78, 129.98, 130.07, 132.24,
132,52, 134.91, 135.14, 147.37, 150.17, 156.99,
192.19.
Maldl-MS (OT/z) ; 556 (M^), 557 (M-"+l).
48
Chapter II, Section 1
Elemental Analysis
Calculated
Found
Calculated for molecular foraiula
C29H22B1N3 O2 S .
C, 62.59; H, 3.98; N, 7.55.
C, 62.64; H, 4.01; N, 7.52%.
2-{S-f(4-Bihenyl(}xy)methylj-4~(phenyl)~4H~l,2,4~triazol~3-yltkio}~l-(4~mtmphenyl)
ethanone (68) Yield: 67 %; Light yellow crystals, m.p, 170-172 "C, Rf = 0.37 (n-
hexane : ethyl acetate; 4;6).
A n
/ / \ V _ o ^ N ^ .
.-1IR (KBr) cm’
’H NMR (400 MHz, CDCb)
NMR (100 MHz, CDCI3)
TOF-MS {m/zd
Elemental Analysis
Calculated
Found
w //
; 2924, 1689, 1525, 1192.
: 5 4.92 (s, 2H, S-CH2), 5.10 (s, 2 H, O-CH2), 6.95
(d, J = 6.3 Hz, 2H Ar-H), 7.30 (t, J = 5.4 Hz,
IH, Ar- H), 7.39-7.42 (m,4H, Ar-H), 7.47 (d, J
= 6.6 Hz, 2H, Ar-H), 7.51 (d, J = 6.3Hz, 2H,
Ar-H), 7.54-7.56 (m, 3H, Ar-H), 8.22 (d, J = 6.6
Hz, 2H, Ar-H), 8.34 (d, 6 .6Hz, 2H, Ar-H).
: 5 39.86, 59.92, 115.17, 124.09, 126.78, 126.92,
128.27, 128.78, 129.72, 130.03, 130.53, 132.43,
134.97, 139.75, 140.46, 150.73, 152.22, 156.95,
191.88.
: 522 (M"'), 523 (M V I).
: Calculated for molecular formula C29H22N4O4S.: C, 66.65; H, 4.24; N, 10.72.
; C, 66.69; H, 4.23; N, 10,6%.
49
Chapter II, Section I
3-(Phenylethyithio)-5-f(4~hiphenyloxy)methyiJ~4-phenyl-4ff-l,2,4~triazoie (69)
Yield: 70%; White crystals, ni.p. 132-134 "C, R|- ~ 0.52 (w-hexane ; ethyl acetate;
4:6),
IR (KBr)
’H NMR (400 M H 2I, CDCia)
13C NMR (100 MHz, CDCI3)
MaMI-MS (m/z)
Elemental Analysis
Calculated
Found
2907,1541, 1523, 1107.
5 3.08 ( t , / = 7.2 Hz, 2H), 3.50 (t, J= 7.8 Hz, 2H
S-CHa), 5.10 ( 2H, s, O-CH2), 6.96 (d, J = 9,0
Hz, 2H,Ar-H), 7.49-7.52 (m, 6H, Ar-H), 7.18
7.25 (m, 6H, Ar-H), 7.31-7.34 (m, 3H, Ar-
H),7.40 (t, 7.2 Hz, 2H, Ar- H).
5 35.86, 36.74, 59.84, 115.16, 123.89, 124.24,
126.88, 128.77, 128.76, 128.79, 129.98, 130.04,
130.34, 130.43, 130.45, 134.91, 142.43, 145.56,
148.34, 148.79, 156.88.
463 (M '), 464 (M"'+1).
Calculated for molecular fonnula C29H2SN3OS.
C, 75.13; H, 5.44; N, 9.06.
C, 75.17; H, 5.42; N, 9.09%.
3-(4-Nitrophenylethylthio)~5-[(4’-biphenyloxy)methyl]'-4-phenyl-4H-l,2,4-
triazole(70) Yield; 74%; Light yellow crystals, m.p. 138-140 “C, Rf = 0.48 (n-hexane
: ethyl acetate; 4:6).
IR (KBr) cm ' : 2907, 1525, 1128.
NM R (400 MHz, CDCI3) : 5 3.12 (t, / = 8.0 Hz, 2H), 3.74 (t, J = 8.4 Hz,
2H), 4.53 (s, 2H, S- CH2), 5.09 (s, 2H, O-CH2),
50
Chapter II, Section I
13C NMR (100 MHz, CDCI3)
Maldl-MS (m/z)
Elementai Analysis
Calculated
Found
6.94 (d, J = 5,7 Hz, 2H, Ar-H), 7.23 (d, J = 6.6
Hz, 2H, Ar-H), 7.30 (t, J = 5.4 Hz, IH, Ar-H),
7.41 (d, J = 7.2 Hz, 2H, Ar-H), 7,47-7.56 (m,
7H, Ar-H), 8.12 (d, / = 6.0 Hz, 2H, Ar-H).
5 35.87, 59.87, 115.17, 123.82, 126.88, 128.77,
129.45, 130.45, 130.11, 130.43, 135.92, 140.45,
144.24, 147.43, 148.45, 156.90.
508 (M"), 509 (M^+1).
Calculated for molecular formula C29H24N4O3S.
C, 68.49; H, 4.76; N, 11.02.
C, 68.55; H, 4.74; N, 11.05%.
3-(4~Nitrophenylmethylthio)-5-[(4-biphenyloxy)methyl]-4'-phenyl-4H-l,2,4-tnazole
i l l ) Yield: 72%; Light yellow crystals, m.p. 136-138 "C, R r= 0.49 (^-hexane : ethyl
acetate; 4:6),
-IIR (KBr) cm'
‘H NMR (400 MMx, CDCI3)
‘• C NMR (100 MHz, CDCI3)
Maldi-MS (m/z)
Elemental Analysis
Calculated
Found
isT/■oO
: 2907, 1516, 1108.
: 5 4.44 (s, 2H, S-CH2), 5,04 (s, 2 H, O-CH2), 6.86
(d, 8.4 Hz, 2H, Ar-H), 7.06-7.14 (m, 5H, Ar-
H), 7.22 (t, 7 = 5,8 Hz, IH, Ar-H), 7.33 (t, J =
7.6 Hz, 2H, Ar-H), 7.38-7.46 (m, 6 H, Ar-H),
8.04 (2H, d, 8,4 Hz, Ar-H).
: 5 36.88, 59.91, 114.98, 115.10, 115.12, 123.31,
126.29, 126.88, 128.80, 129.91, 130.18, 130.29,
130.88, 130.46, 131.26, 134.90, 137.75, 146.26,
146.56, 155.74, 156.86.
494 (M'"), 495 (M'^+l).
Calculated for Molecular formula C28H22N4O3S;
C, 68,00; H, 4.48; N, 11.33.
C, 68.10; H, 4.51; N, 11.36%.
51
Chapter II, Section 1
3~(Methylthio)S-[(4-biphenyloxy)methyl]-4-phenyl-4H-l,2,4~‘triazole (72) Yield; 6 8%; White crystals, m.p. 168-170 "C, R r= 0.28 (n-hexane : ethyl acetate; 4:6).
IR (KBr) cm‘‘
‘h NMR (300 MHz, CDCI3)
!3 C N M R (75 MHz, CDCI3)
Maldi-MS (m/z) Elemental Analysis
Calculated
Found
: 2912, 1541, 1524, 1109.
: 8 2.70 (s, 3H, S-CII3), 5.10 (2 H, s, O-CH2), 6.94
(d, ./= 8.4 Hz, 2H), 7.30-7.52 (m, 12H, Ar-H).
: 6 14.59, 59.99, 115.20, 126.77, 126.86, 126.97,
128.22, 128.75, 129.83, 130.23, 132.83, 134.85,
140.53, 151.86, 154.26, 157.05.: 373 (M"), 374 (M‘"+l).
: Calculated for molecular formula C22H19N3OS.: C, 70.75; H, 5.13;N, 11.25.
: C, 70.69; H, 5.16; N, 11.29%.
3~(Ethylthio)-5-[(4-biphenyloxy)tnethyl]-4-phenyl-4H-l,2,4~triazole (73) Yield:
74%; White crystals, ni.p. 174-176 "C, Rf = 0.26 (n-hexane : ethyl acetate; 4:6).
Q N-// N— < IIN
Ha
-1IR (KBr) cm
NMR (300 MHz, CDCI3)
; 2907,1541, 1523, 1108.
: 5 1.42 (t, J = 5.6 Hz, 3H, -CH3), 3.29 (q, J =
5.4 Hz, 2H, S-CH2-), 5.14 (s, 2H, O-CHs), 6.88
(d, / - 8.3 Hz, 2H), 7.21 (t, J = 7.8 Hz, 2H),
7.44-7.54 (m, 5H, Ar-H), 7.26-7.42 (m, 5 H,
Ar-H).
52
Chapter II, Section 1
13'C NMR (75 MHz, CDCI3)
Maldi-MS (m/z)
Elemental Analysis
Calculated
Found
5 14.60, 29.68, 59.92, 115.06, 116.72, 117.04,
126.74, 126.89, 128.25, 128.68, 129.02, 129.19,
135.06, 140.44, 156.92.
387 (M"‘), 388 (M'^+1).
Calculated for molecular formula C23H21N3OS.
C, 71.29; H, 5.46; N, 10.84.
C, 71.27; H, 5.45; N, 10.32%.
2-f5-{(4-Biphenyioxy)methyl}-4-(4-methoxyphenyl)-4H-l,2,4~triazol~3-ylthio]-l-(4-
hrom oph enyl)eth an on e (74) Yield: 70 %; White crystals, m.p, 156-158 "C, Rf = 0.18
(n-liexane : ethyl acetate; 4:6).
Br
CH.
IR (KBr) cm'V
‘H NMR (CDCI3)
'^C NMR (CDCI3)
126.98,
Maldi-MS {m/z)
Elemental Analysis
Calculated
Found
: 2907, 1686, 1605, 1536, 1105.
: 6 3.88 (s, 3H, O-CH3), 4.85 (s, 2H, S-CH2), 5.15
(s, 2H, O-CH2), 6.90 (d, 6.6 Hz, 111, Ar-H),
7.01 (d, 6.7 Hz, 2H, Ar-H), 7.18- 7.24 (m,
2H, Ar-H), 7.28 (t, J - 5.5 Hz, IH, Ar-H), 7.37-
7.51 (m, 6H, Ar-H), 7.48-7.53 (m, 4H, Ar-H).
; 5 37.58, 56,32, 60.09, 114.94, 122.48, 126.80,
127.22, 128.56, 128.74, 128.84, 128.96, 129.08,
130.65, 132.24, 134.81, 135.89, 140.34, 147.34,
150.25, 156.84, 160.65, 191,45.
: 586 (M^), 587 (M'^'+l).
: Calculated for molecular formula
C3oH24Bi'N303S.
: C, 61.44; H, 4.12; N, 7.16.
:C , 61.48; H, 4.09; N, 7.13%.
53
Chapter II, Section I
2-[5-{(4~Biphenyloxy)methyl)-4-(4-methoxyphenyl)-4H-l,2,4~triazol-3~ylthioJ~l-(4-
chiorophenyl)ethati0ne (75) Yield: 79%; White crystals, ni.p. 153-155 ”C, Rf = 0.21
(«"hexane : ethyl acetate; 4:6).
IR (KBr) cm“‘
'H NMR (400 MHz, CDCI3)
13C NMR (lOOMIfe, CDCI3)
Malcii-MS ( m/z^
Elemental Analysis
Calculated
Found
Cl
C I-I3
: 2907, 1655, 1605, 1239, 1093.
: 5 3.85 (s, 3 H,0 -CH3), 4.91 (s, ZH^S-CHa), 5.12
(s, 2H, O-CH2), 6.96 (d, J = 8,4 Hz, 2H, Ar-H),
7.00 (d, J = 9 Hz, 2H, Ar-H), 7.25-7.32 (m, 5H,
Ar-H), 7.40 (t, y - 7.5 Hz, 2H, Ar-H),7.90 (d, J
= 8.4 Hz, 2H, Ar-H), 7.46-7.52 (m, 4H, Ar-H).
: 5 38.03, 55.35, 60.12, 114.94, 115.27, 126.81,
127.12, 128.08, 128.57, 128.99, 129.03, 130.73,
132.24, 134.81, 135.87, 137.87, 140.21, 147.35,
150.15, 156.84, 160.69, 190.24.
: 542 (M^), 543 (M V I).
; Calculated for molecular formula
C30H24CIN3O3S.:C , 66.47; H, 4.46; N, 7.75.
: C, 66.53; H, 4.44; N, 7.69%.
2-[5~{(4-Biphenyloxy)methyl}-4-(4-methoxyphenyl)-4H~l,2,4-triazol-3-ylthioJ-l-’
(phenyl)ethanone (76) Yield: 82%; White crystals, m.p. 150-152 °C, Rf = 0,22 {n-
hexane : ethyl acetate; 4:6).
CH-,
54
Chapter //, Section 1
fR (KBr) cm '
'H NMR (400 MHx, CDCI3)
'-■'C NMR (lOOMHz, CDCI3)
Maldi-MS (w/^)
Elemental Analysis
Calculated
Found
2907, 1686, 1541, 1102.
5 3.85 (s, 3H, O-CHj), 4.92 (s, 2H, S-CH2), 5.10
(s, 2H, O-CH2), 6.96 (d, . /= 8,4 Hz, 2H, Ar-H),
7.25-7.32 (m, 6H, Ar-H), 7.40 (t, J = 7.5 Hz,
2H, Ar-H), 7.40-7.46 (m, 6H, Ar-H), 7.91 (d, ./
= 8.4 Hz, 2H, Ar-H).
5 36.13, 56.14, 60.24, 114.94, 115.12, 126.79,
127.12, 128.09, 128.56, 128.92, 129.05, 130.76,
132.27, 133.20, 134.83, 135.87, 137.26, 147.37,
150.17, 156.81, 160.69, 190.20.
507 (M^), 508 (M'^+l).
Calculated for molecular formula C30H25N3O3S. C, 70.98; H, 4.96; N, 8.28
C, 71.02; H, 4.95; N, 8.25%.
2-[5-{(4-Biphenyloxy)methyl}~4-(4-methoxyphenyl)-4H-l,2,4-triazol-3-ylthio]-l-'(4
methoxyphenyl)ethanone (77) Yield; 78 % ; White crystals, m.p. 157-159 ”C, Rf = 0.21 (n-hexane : ethyl acetate; 4:6).
-iIR (KBr) cm'
’H NMR (400 MHz, CDCI3)
'•C NMR ( 1 0 0 MHz, CDCb)
: 2907, 1687, 1586, 1087.
: 5 3.87 (s, 6H, O-CH3), 4.92 (s, 2H, S-CH2), 5.15
(s, 2H, O-CH2), 6.88-7.08 (m, 6H, Ar-H), 7.31
(t, J = 5.8 Hz, IH, Ar-H), 7.41 (t, J = 7.5 Hz,
2H, Ar-H), 7.46-7.52 (m, 4H, Ar-H), 7.67 (d, /
= 8.9 Hz, 2H, Ar-H), 7.99 (d, J = 9.0 Hz, 2H,
Ar-H).
: 5 37,10, 55.63, 60.25, 114.92, 114.94, 115.13,
126.80, 127,10, 127,13, 128.56, 128.92, 129.07,
55
Chapter II, Section 1
Maldi-MS im/z)
Elenientsil Analysis
Calculated
Found
129.96, 130.96, 134.54, 135.14, 147.38, 150.19,
160,68, 156.82, 164.56, 191.94,
537 (M^), 538 (M U i ).
Calculated for molecular fonnula C31H27N3O4S. C, 69.25; H, 5.06; N, 7.82,
C, 69.30; H, 5.02; N, 7.79%,
2-[5-((4-Biphenyloxy)methyl}~4-(4-
phenylphenyl)ethanone (78) Yield:
(«"hexane : ethyl acetate; 4:6).
■methoxyphenyl)~4H-l,2i4-triazoi-3-ylthiaJ~l~(4-
: 84 %; White crystals, ni.p.168-170 "C, Rf = 0.20
IR (KBr) cm'*
' H NMR (400 MHz, CDCb)
NMR (100 MHx, CDCI3)
ES-MS (m/z)
Elemental Analysis
Calculated
Found
2907,1688,1586,1 109.
d 3.87 (s, 3H, O-CH3), 4.92 (s, 2 H, S-CH2), 5.15
(s, 2H, O-CH2), 6.88-7.10 (m, 4H, Ar-H), 7.31
(m, 2H, Ar-H), 7.41 (t, J = 7.5 Hz, 2H, Ar-H),
7.48-7.50 (m, lOH, Ar-H), 7.68 (d, J = 8.8 Hz,
2H, Ar-H), 8.09 (d, J= 9.0 Hz, 2H, Ar-H).
5 37.10, 56.64, 61.35, 114.92, 114.94, 115.13,
126.80, 127.10, 127.13, 128.56, 128.92, 129.07,
129.96, 130.96, 134.54, 135.14, 147.38, 150.19,
156.88, 160.68, 164.59, 192.14.
584 (M"'+1), 585 (M'^+2).
Calculated for molecular formula C36H29N3O3S.
C, 74.08; H, 5.01; N, 7.20.
C, 74.12; H, 5.02; N, 7.22%.
56
Chapter n, Section 1
3~(4-Nitrflphenylmethylthlo)-5-[(4-hiphettyloxy)methyl]~4(4-methoxyphenyl)~4H-
h2,4-triazole (79) Yield; 72 %; White crystals, m.p. 173-175 "C, Rf - 0.25 (n-
hexane : ethyl acetate; 4:6).
IR (ICBr) cm‘‘
NMR (400 M Hz, CDCh)
NMR (100 MHz, CDCI3)
ES-MS (m/z) Elemental Analysis Calculated Found
NO2
CH3
2914, 1687, 1556, 1091.
5 3.87 (s, 3H, O-CH3), 4.42 (s, 2H, S-CH2), 5.12
(s, 2H, O-CH2), 6 .86-6.88 (m, 4H, Ar-H), 7.06-
7.14 (m, 2H, Ar-H), 7.22 (t, J - 5.8 Hz, IH, Ar-
H), 7.33 (t, 7.6 Hz, 2H, Ar-H), 7.40-7.44 (m,
6H, Ar-H), 8.04 (d, 8.4 Hz, 2H, Ar-H).
5 36.88, 59.91, 64.34, 164.72, 114.98, 115.16,
114.82, 123.34, 126.31, 126.88, 128.82, 129.94,
130.21, 130.29, 130.84, 131.36, 135.12, 137.75,
146.16, 146.58, 155.74, 156.88.
525 (M^+1), 526 (M'^+2)Calculated for molecular formula C29H24N4O4S. C, 66.40; H ,4 .61;N , 10.68 C, 66.42; H, 4.62; N, 10.67%.
3-(phenylethylthio)~5-[(4-biphenyloxy)methyl]-4(4-methoxyphenyl)-4H-l,2,4-triazole (80) Yield: 78 %; White crystals, m .p.144-146 "C, Rf = 0.32 (n-hexane : ethyl acetate; 4:6),
-1IR (KBr) cm"
NMR (400 MHz, CDCI3)
2916, 1689, 1562, 1096.
5 3.08 (2H, t, J - 7.2 Hz), 3.50 (t, J = 7.8 Hz,
2H, S-CH2), 3.86 (s, 3H, O-CH3), 5.10 ( 2 H, s,
57
Chapter II, Section 1
13C NMR (100 MHz, CDCI3)
Maldl-MS {m/7) Elemental Analysis
Calculated
Found
O-CH2), 6.88-6.96 (m, 4H, Ar-H), 7.16-7.24 (m,
4H, Ar-H), 7.32-7.34 (ra, 2H, Ar-H), 7.40 (t, . / -
7.2 Hz, 2H, Ar-H), 7.48-7.54 (m, 6 H, Ar-H).
: 8 35.86, 36.74, 56.62, 59.84, 115.18, 123.89,
124.22, 126.88, 128.70, 128.74, 128.89, 129.96,
130.04, 130.24,130.40,130.48, 134.92, 142.42,
145,46, 148.24, 148.74,156.88,164.12.
; 493 (M''), 494 (M'"+l),
: Calculated for molecular formula C30H27N3O2S.
: C, 73.00; H, 5.51; N, 8.51.
: C, 73.08; H, 5.52; N, 8.49%.
2-f5-{(4-BiphenyIoxy)methyl}~4-(4-flourophenyl)-4H-l,2,4-triazol~3-ylthioJ~l- 4bromophenyl) ethanone (81) Yield: 72%; White crystals; m,p. 160-162”C, Rf =
0.39 (/?-hexane : ethyl acetate; 4:6).
Br
IR (KBr) cm-‘
‘H NMR (400 MHz, CBCI3)
NMR (100 MHz, CDCI3)
Maldi-^MS (m/z)
Elemental Analysis
Calculated
Found
2907, 1684, 1605, 1108.
6 4.92 (s, 2H, S-CH2), 5.08 (s, 2H, O-CH2),
6.92-7.90 (m, 17 H, Ar-H).
5 36.79, 59.85, 114.97, 115.15, 126.23, 126.79,
126.87, 128.78, 130.09, 130.41, 130.46, 130.76,
130.87, 131.29, 134.89, 137.24, 145.58, 145.69,
156.87, 162,98, 192.34.
574 (M""), 575 (M'^+l).
Calculated for molecular fomiula
CzoHsiBrFNaOzS.
C, 60.63; H, 3,68; N, 7,31
C, 60.67; H, 3.69; N, 7.32%.
58
Chapter II, Section J
2-fS~{(4-Biphenyloxy)methyi}-4~(4’-fluorophenyl)-4H-I,2,4-triazol-3~ylthio]~l~
(phenyl) ethanone (82) Yield; 74%; White crystals, m.p,156-158“C, Rf = 0.29 (n~
hexane : ethyl acetate; 4:6).
IR (KBr) cm"'
’H NMR (300 MH^, CDCI3)
13C NM R (100 MHz, CDCI3)
M aldi-M S (m/z)
Elemental Analysis
Calculated
Found
: 2933, 1698, 1515, 1105.
: 8 4.92 (s, 2H, S-CH2), 5.08 (s, 2H, O-CH2), 6.93
(d, J = 6.3Hz, 2H, Ar-H), 7,29 (t, J = 5.4 Hz,
IH), 7.41 (t, J = 8.4Hz, 2H, Ar-H), 7.37-7.41
(m, 4H, Ar-H), 7.46-7.51 (m, 5H, Ar-H), 7.63
(d, y - 6 Hz, 2H),7.89 (d, J= 6.3 Hz, 2H).
: 5 37.03, 58.97, 114.79, 115.13, 126.25, 128.77,
129.89, 130.10, 130.12, 130.45, 130.67, 130.87,
130.88, 133.89, 134.34, 136.15, 137.15, 145.27,
145.63, 156.78, 162.97, 191.36.
; 495 (M"'), 496 (M’ +l).
: Calculated for molecular formula
C29H22GIFN3O2S.
: C, 70.29; H, 4.47; N, 8.48.
: C, 70.28; H, 4.43; N, 8.50%.
2-[5-((4~biphenyloxy)methyl}-4~(4-fluorophenyl)-4H~l,2,4-triazol-3~ylthio]-l-(4-
methoxyphenyl) ethanone (83) Yield: 85%; White crystals, m.p.172-174 "C, Rf =
0.30 (n-hexane : ethyl acetate; 4:6).
CHi
59
Chapter II, Section 1
IR (KBr) cm'*
u C NMR (100 MHz, CDCI.,)
MaMi-MS (w/z)
Calculated
Found
; 2907, 1686, 1605, 1093.
; 5 3.87 (s, 3H, O-CH3), 4.91 (s, 2H, S-CHa,),
5.14 (s, 2H O-CH2.), 6.87-7.89 (m, 17 H, Ar-H).
: 5 37.47, 55.87, 58.98, 114.15, 114.78, 114.96,
115.14, 126.26, 128.00, 129.87, 130.11, 130.12,
130.15, 130.26, 130.89, 133.91, 145.27, 146.00,
156.87, 161.67, 162.97, 192.05.
; 525 (M'"), 526 (M^+1).
: Calculated for molecular fonnula
C30H24FN3O3S.
: C, 68.56; H, 4.60; N, 7.99.
:C , 68.60; H, 4.58; N, 8.01%.
3-(Phenylethylthia)-'5-[(4-biphenyloxy)methylJ-4-(4-fluorophenyl)-4H-l,2,4-triazoie
(84) Yield: 62%; White crystals, m.p. 144-146 "C, Rf = 0.41 (n-hexane : ethyl
acetate; 4:6),
-1IR (KBr) cm'
NM R (300 MHz, CDCI3)
NM R (100 MHz, CDCI3)
Maldi-MS (m/z)
: 2907, 1520, 1517, 1108.
; 8 3.08 (t, 8.1 Hz, 2H, -CH2-), 3.50 ( t ,J = 7.5
Hz, 2H, S-CH2), 5.08 (s, 2H, O-CH2), 6.96 (d, J
= 9.0 Hz, 2H, Ar-H), 7.18- 7.34 (m, 8H, Ar-H),
7.40 (t, / = 7.8 Hz, 2H, Ar-H), 7.49-7.52 (m,
6H, Ar-H).
5 37.49, 55.58, 59.12, 114.56, 114.78, 123.07,
127.99, 129.17, 129.85, 130.03, 130,08, 130.25,
130.45, 130.98, 134.79, 135.14, 145.14, 147.18,
149.00,156.98,161.69.
481 (M""), 482 (M M ).
60
Chapter II, Section 1
Elemental Analysis
Calculated
Found
; Calculated for molecular formula C29H24FN3OS.
: C, 72.33; H, 5.02; N, 8.73,
: C, 72.29; H, 5.04; N, 8.69%.
3~(4~Nitrophenylmethylthio)~5{(4~hiphenyloxy)methyll-4-(4~fluwophenyl)~4H-
1,2,4-triazole (85) Yield." 6 8 %; Light yellow crystals; m ,p.l68-172 "C, Rf = 0.29 {n-
hexane ; ethyl acetate; 4:6).
NO2
IR (KBr) cnf*
’H NMR (400 MHz, CDCI3)
NM R (100 MHz, CDCI3)
Maldi-M S {m/i)
Elemental Analysis
Calculated
Found
: 2916, 1524, 1106.
; S 4.45(s, 2H, S-CH2), 5.01 (s, 2H, O-CH2), 6.87
(d, J - 8.4 Hz, 2H, Ar-H), 7.08-7.16 (m, 4H, Ar-
H), 7.23(t, J - 5.7 Hz, IH, Ar-H), 7.33 (t, J =
7.6 Hz, 2H, Ar-H), 7.40-7.49 (m, 6H, Ar-H),
8.04 (d, 8.4 Hz, 2H, Ar-H).
: 5 36.86, 59.89, 114.97, 115.14, 123.27, 126.27,
126.89, 128.79, 129.89, 130.12, 130.27, 130.48,
130.89, 131.27, 134.91, 137.73, 146.23, 146.57,
155.76, 156.89, 162.97,
: 512 (M’"), 513 (M‘*'+l).
: Calculated for molecular fonnula
C28H21FN4O3S.
: C, 65.61; H, 4.13; N, 10.93.
: C, 65.67; H, 4.09; N, 10.90%.
61
Chapter II, Section 1
3~(EthyltM0)S[(4~biphemyloxy)methylj-4-(4-fluor0phenyl)~4H-lf2,4-triazole (8 6 )
Yields 6 8 %; White crystals; m ,p.l42-144 ”C, Rf = 0.25 («-hexane ; ethyl acetate;
4:6).
IR (KBr) cm"'
’H NMR (300 MHz, CDCIj)
NM R (75 MHz, CDCI3)
M aldi-M S {m/z)
E lem ental Analysis
Calculated
Found
2920, 1526, 1108.
5 1,42 (t, J = 5 .6 Hz, 3H, -CH3), 3.29 (q, ^ - 6.4
Hz, 2 H, S-CHz"), 5.11 (s, 2H, O-CH2), 6.95 (d,
.7 -8 .1 Hz, 2H), 7.20 (t, . / - 7.8 Hz, 2H), 7.30-
7.43 (m, 5 H, Ar-H),7.46-7.52 (m, 4H, Ar-H).
5 14.66, 29.68, 59.90, 115.07, 116.76, 117.07,
126.75, 126.89, 128.27, 128.73, 129.07, 129.19,
135.01, 140.42, 156.86, 164.94.
405 (M'"), 406 (M^+1).
Calculated for molecular formula C23H20FN3OS. C, 68.13; H, 4.97; N, 10.36.
C, 68.27; H, 4.95; N, 10.32%.
3-(Methylthio)-5[(4~biphenyloxy)methylJ~4~(4~fluorophenyl)-4H-l,2,4-triazole (87)
Yield: 69 %; Light yellow crystals; m.p.140-142 "C, Rf = 0.24 («-hexane : ethyl
acetate; 4:6)
-IIR (KBr) cm’*H NMR (300 MHz, CDCI3)
; 2916, 1524, 1106.
: 5 2.70 (s, 3H, S-CH3), 5.09 (s, 2H, O-CH2), 6,95-
7.49 (m, 13 H).
62
Chapter II, Section 1
13C NMR (75 MHz, CDCI3)
Maldl^MS (m/z)
Elem ental Analysis
Calculated
Found
5 14.54, 29.72, 59.95, 115.11, 116.81, 117.11,
126.78, 126.92, 128.29, 128.71, 128.77, 129.01,
129.14, 134.96, 140.47, 151.92, 154.43, 156.93,
161.59, 164.92.
391 (M‘"), 392 (M V i).
Calculated for molecular formula C22H 18FN3OS.
C, 67.50; H, 4.63; N, 10.73.
C, 67.57; H, 4.66; N, 10.76%.
2-[5-((4-Biphenyloxy)methyl}-4-(4~chlorophenyl)-4H~l,2,4-triazol-3-ylthio]~l-(4-
brotftophenyl)ethanone (88) Yield: 6 8%; white crystals, m.p.154-156 "C, Rf = 0.37
(«!-hexane ; ethyl acetate; 4;6)
IR (KBr) cm'*
‘H NMR (400 MHz, CDCI3)
NM R (100 MHz, CDCI3)
Maldi-M S {m/z:)
E lem ental Analysis
Calculated
Found
: 2912, 1683, 1607, 1584, 1093.
: 5 4.92 (s, 2 H, S-CH2), 5.12 (s, 2H, O-CH2), 6.94
(d, J = 6.3 Hz, 2H), 7.29 (t, J = 5.7 Hz, IH),
7.34 (d, J = 6.3 Hz, 2H), 7.39 (t, J = 5.7 Hz, 2H,
Ar-H), 7.46-7.51 (m, 6H, Ar-H), 7.62 (d, J =
6.0 Hz, 2H, Ar-H),7.88 (d, J-= 6 Hz, 2H, Ar-H).
: 5 35.87, 59.96, 115.18, 126.88, 126.92, 127.65,
128.76, 128.82, 128.87, 129.27, 129.35, 130.06,
130.26, 130.85, 134.76, 135.17, 142.00, 145.25,
148.24, 156.95, 192.64.
: 590 (M'"), 591(M'"+1).
: Calculated for molecular formula
C29H2iClBrN302S.
: C, 58.94; H, 3.58; N, 7.11.
: C, 59.02 H, 3.59; N, 7.08%.
63
Chapter II, Section 1
2-[5-((4-Biphenyloxy)methyl}-4-(4-chlorophenyl)-4H-l,2,4~trlazol-3-ylthioJ~l-(4~ mtrophenyl)ethanone (89) Yield: 72%; light yellow crystals, m,p,183-184 ”C, Rf =
0.35 (n-hexane : ethyl acetate; 4:6).
‘-■’C NM R (100 MHz, DMSO-df,)
IR (KBr) cm"' : 2905, 1684, 1605, 1525, 1320, 1093.
'H NMR (400 M H z, CDCb) : 8 4.91 (s, 2H, S-CH2), 5.10 (s, 2H, O-CH2), 6.95
(d, J = 6.3 Hz, 2H, Ar-H), 7.31 (t, J = 5.7 Hz,
IH, Ar-H), 7.37(d, 6.6 Hz, 2H, Ar-H), 7.41
( t ,y = 5.7Hz, 2H), 7.47-7.53 (m, 6H, Ar-H),8.20
(d, J - 6.6 Hz, 2H, Ar-H), 8.34 (d, J - 6.6 Hz,
2H, Ar-H).
5 36.75, 59.87, 115.10, 124.09, 126.79, 126.97,
128.31, 128.78, 130.28, 130.92, 131.12, 135.17,
136.00, 139.72, 140.41, 147.00, 150.00, 156.81,
191.65.
557 (M^), 558 (M^+1).
Calculated for molecular formula
C29H21CIN4O4S.
C, 62.53; H, 3.80; N, 10.06.
C, 62.62; H, 3.82; N, 9.96%.
2-[5-{(4-Biphenyloxy)methyl}-4-(4-chlorophenyl)-4H-l,2,4-triazol-3-ylthio]~l-(4-
methoxyphenyl) ethanone (90) Yield: 75%; Wliite crystals, m.p. 178-180 °C, Rf =
0.34 (n-hexane : ethyl acetate; 4:6).
Maldi-M S {m/z)
Elem ental Analysis
Calculated
Found
'CH,
IR (KBr) cm' : 2912, 1681, 1607, 1093.
64
Chapter II, Section 1
‘H NMR (400 MHz, CDCI3)
13C NMR (100 MHz, DMSO-dg)
Maldi“MS {m/z)
Calculated
Found
: 6 3.83 (s, 3H, O-CH3), 4.53 (s, 2H, S-CH2),
5.01 (s, 2H, O-CH2), 6.91-6.96 (m, 4H, Ar-H),
7.27-7.33 (m, 3H, Ar-H), 7.46-7.51 (m, 6 H, Ar-
H), 7.61 (d, J = 6.4 Hz, 2H, Ar-H),7.85 (d, J =
6.4 Hz, 2H, Ar-H).
: 5 36.96, 56.24, 60.07, 114.98, 117.56, 126.88,
127.24, 127.85, 128.95, 128.96, 129.57, 129.86,
130.16, 130.26, 131.05, 134.78, 135.16, 145.75,
148.47, 156,34, 156.78, 192.94.
: 542 (M^), 543 (M'^+1).
; Calculated for molecular formula
C30H24CIN3O3S.
: C, 66.47; H, 4.46; N, 7.75.
: C, 66.50; H, 4.47; N, 7.72%.
2-l5-{(4-Biphenyloxy)methyl}-4-(4-chlorophenyl)-4H-l,2,4-triazol~3-ylthio]~l-
(phenyl) ethanone (91) Yield: 68 %; White crystals, m.p.192-194 "C, Rf = 0.36 (n-
hexane : ethyl acetate; 4:6).
IR (KBr) cm '
'H NM R (400 MHz, CDCI3)
Elemental Analysis
Calculated
Found
: 2924, 1680, 1584, 1107.
: 5 4.51 (s^+1).
: Calculated for molecular formula
C29H22CIN3O2S.
:C , 68.03; H, 4.33; N, 8.21.
: C, 68.09; H, 4.30; N, 8.19%.
65
Chapter II, Section 1
2-[5-{(4-Biphenyloxy)methyl}~4~(3~chl0mphenyl)-4H~ly2,4-triazol'-3-ylthio]~l’-(4-
methoxyphenyl) ethanone (92) YJeld; 82%; White Crystals, iti.p. 162-164 “C, Rf =
0,20 («-hexane : ethyl acetate; 4:6).
IR (KBr) cm'*
‘H NMR (400 MHz, CDCI3)
NM R (100 MHz, DMSO-d<;)
ES-MS (w/z)
Elemental Analysis
Calculated
Found
: 2917, 1526, 1109.
: 5 3.88 (s, 3H, O-CH3), 4.93 (s, 2H, S-CH2), 5.11
(s, 2H, O-CH2), 6.94-6.97 (m, 4H), 7.31 (d, J =
6.6 Hz, IH), 7.38-7.53 (m, lOH, Ar-H), 8.01 (d,
J = 8 .7 H z , 2H).
: 5 36.96, 55.89, 60.07, 114.98, 117.56, 126.88,
127.24, 127.85, 128.95, 129.86, 128.96, 129.57,
130.16, 130.26, 131.05, 134.78, 135.16, 146.15,
148.42, 156.38, 156.72, 191.92.
; 542 (M'"+l), 543 (MV2).
: Calculated for molecular foimula
C30H24CIN3O3S.: C, 66.47; H, 4.46; N, 7.75.
: C, 66.58; H, 4.49; N, 7.75%.
2-[5-{(4~Biphenyloxy)methyl}-4-(3-chlorophenyl)-4H-l,2,4-triazol-3-ylthioJ-
1 (phenyl) ethanone (93) Yield: 76%; White Crystals, m.p. 143-145 °C, Rf == 0.31 {n-
hexane ; ethyl acetate; 4:6).
6 6
Chapter II, Section 1
-1IR (KBr) cm
% NMR (400 MHz, CDCI3)
13C NMR (100 MHz, DMSO-dg)
Maldl-MS (m/z)
Elemental Analysis
Calculated
Found
: 2914, 1520, 1108.
: 5 4.98 (s, 2H,S-CH2), 5.12 (s, 2H,0 -CH2>, 6.96
(d, J = 7.9 Hz, 2H), 7.32 (d, J = 7.2 Hz, 2H),
7.39-7.43 (m, 3H, Ar-H), 7.51-7.59 (m, 8H, Ar-
H), 7.62 (t, J = 7.2 Hz, IH, Ar-H), 8.03 (d, J
= 8.1 Hz, 2H).
8 38.12, 60.12, 114.89, 126.83, 127.26, 127.84,
128.66, 128.96, 128.97, 129.19, 129.72, 130.27,
132.75, 131.04, 135.16, 136.45, 145.73, 148.45,
156.42, 191.82.
512 (M'"'), 513 (M '+I).
Calculated for molecular fomnula
C 2 9 H 2 2 C IN 3O 2 S .
C, 68,03; H, 4.33; N, 8.21.
C, 68.58; H, 4.36; N, 8.25%.
2-fS-{(4~Biphenyloxy)methyl}-4-(3~chlorophenyl)-4H~l,2,4~triazol-3-ylthioJ-l~(4~
phenyl-phenyl)ethanone (94) Yield: 80%; White Crystals, m.p. 164-166 "C, Rf
0.41 («-hexane ; ethyl acetate; 4:6).
-IIR (KBr) cm"
‘H NMR (400 MHz, CDCI3)
NMR (100 MHz, CDCI3)
: 2916, 1556, 1108.
: 5 5.0 (s, 2H, S-CH2), 5.12 (s, 2H, O-CH2), 6.96
(d, 8.7 Hz, 2H), 7.28-7.50 (m, 14H), 7.58 (d,
J = 6.9 Hz, 2H), 7.72 (d, J - 8.1 Hz, 2H), 8.10
(d, J= 8 .4 H z ,2 H ).
; 5 37.10, 56.64, 61.33, 114.90, 114.92, 115.10,
126,80, 127.06, 127.10, 128.54, 128.90, 129.17,
67
Chapter II, Section 1
Maldi-MS (/k/ x)
Elemental Analysis
Calculated
Found
129.94, 130.94, 134.51, 135.14, 146.36, 150.19,
156.84, 160.68, 163.88, 192.14,
: 588 (M^), 589 (M"’+2).
: Calculated for molecular formula
C35H26CIN3O2S.
; C, 71.48; H, 4.46; N, 7.14.
C, 71.62; H, 4.51; N, 7.19%.
2-f5~{(4"Biphenyloxy)methyl}-4~(4~mtrophenyl)"4H-l,2,4-triazol~3-ylthw]-l~(4-
methoxyphenyl)ethanone (95) Yield: 78%; Light yellow crystals; m .p .l98-200 "C,
R f - 0 . 19 («-hexane : ethyl acetate; 4:6).
.-sIR (KBr) cm'
'H NM R (400 M fe , CDCI3)
NM R (100 MHz, CDCI3)
M aldi-M S (m/z)
Elem ental Analysis
Calculated
Found
: 2912, 1686, 1518, 1528, 1105.
; 5 3.89 (s, 3H, O-CH3), 4.92 (s, 2H, S-CH2), 5.12
(s, 2H, O-CH2), 6.98 (d, J = 6 .6 Hz, 2H, Ar-H),
7.28-7.52 (m, 9H, Ar-H), 7.66 (d, J = 8.9 Hz,
2H, Ar-H), 7.98 ( d ,J = 9.0 Hz, 2H, Ar-H),
8.42 (d, J = 8.7 Hz, 2H, Ar-H).
: 8 35.79, 56.82, 59.84, 115.15, 122.36, 124.06,
126.74, 128.30, 128.65, 130.25, 130.64, 135.74,
135.84, 138.54, 141.44, 147.32, 150.10, 156.86,
192.22.
552 (M^), 553 (M ‘ + 1).
Calculated for molecular formula C30H24N4O5S.
C, 65.20; H, 4.38; N, 10.14
C, 65.22; H, 4.37; N, 10.12%.
6 8
Chapter II, Section 1
2-[5-{(4-Biphenyloxy)methyl}-4-(4-mtrophenyl)~4H~l,2,4~triazol-3-ylthi0]~l-(4-
chlorophenyl)ethanone (96) YieM'o 76%; Light yellow crystals, m .p. 158-160 ‘'C, Mf
= 0.33 («-hexane ; ethyl acetate; 4:6).
Cl
IR (KBr) cm”
'H NMR (300 MHz, CDCI3)
‘- C NMR (100 MHz, CDCI3)
Maldi-MS {m/z}
Elemental Aiaalysis
Calculated
Found
: 2912, 1686, 1604, 1556, 1105.
: 5 4.91 (s, 2H, S-CH2), 5.14 (s, 2H, O-CH2), 6.92
(d, J = 8.7 Hz, 2H, Ar-H), 7.32 (d, J - 7.5 Hz,
2H), 7.41 (t, 7.5 Hz, IH, Ar-H), 7.47-7.68
(m, 8 H, Ar-H), 7.87 (d, 8.4 Hz, 2H, Ar-H),
8.42 (d, J - 8.7 Hz, 2H, Ar-H).
: 6 34.56, 59.96, 79.82, 115.23, 122.54, 124.19,
126,83, 128.36, 128.42, 128.59, 130.32, 130.71,
135.81, 135.86, 134.59, 141.52, 148.14, 150.24,
156.92, 191.26.
: 557 (M'") 559 (M^+1).
: Calculated for molecular formula
C29H21CIN4O4S.: C, 62.53; H, 3.80; N, 10.06.
: C, 62.50; H, 3.81; N, 10.08%.
2~[5-{(4-Biphenyloxy)methyl}-4-(4-nitrophenyl)-4H-l,2,4~triazol-3-ylthio]-l~
(phenyl)ethanone (97) Yield: 66%; Light yellow crystals, m.p. 168-170 "C, Rf =
0.27 (n-hexane : ethyl acetate; 4:6).
//^"NAO N - ^
IR (KBr) cm"*H NMR (400 MHz, CDCI3)
\ JO2N
: 2907,1686, 1515, 1524, 1093.
: 5 4.91 (s, 2 H, S-CHz), 5.14 (s, 2H, O-CH2), 6.96 (d, J = 6.6 Hz, 2H, Ar-H), 7.28-7.52 (m, lOH,
69
Chapter II, Section 1
'•"'C NMR (100 MHz, CDCI3)
Maldl-M S im/z)
Elemental Analysis
Calculated
Found
Ar-H), 7.66 (d, 8.9 Hz, 2H, Ar-H), 7.98 (d,J = 9.0 Hz, 2H, Ar-H), 8.41 (d, J = 8.7 Hz, 2H, Ar-H).5 35.00, 59.86, 79.72, 115.17, 122.34, 124.09, 126.79, 128.31, 128.32, 128.65, 130.27, 130.68,135.76, 135.86, 138.56, 141.46, 147.34, 150.12,156.88, 191.24.
522 (M"‘), 523 (M"'+1).
Calculated for molecular formula C29H22N4O4S.
C, 66.65; H, 4.24; N, 10.72
C, 66.72; H, 4.26; N, 10.68%.
2~[5-{(4-Biphenyloxy)methyl}-4-(4-mtrophenyl)~4H~l,2,4~triaz.ol-3~ylthioJ~l-(4-
phenylphenyl)ethanone (98) Yield; 70%; Light yellow crystals, m .p ,175-177 "C, Rf
= 0.35 (n-hexane : ethyl acetate; 4:6).
IR (KBr) cm'
’H NMR (300 MHz, CDCI3)
NMR (100 MHz, CDCI3)
Maldi-MS (m/z)
Elemental Analysis
Calculated
Found
2907, 1671, 1601, 1558, 1093.
5 5.00 (s, 2 H, S-CH2), 5.15 (s, 2H, O-CH2), 7.31
(t, 5.7 Hz, IH, Ar~H), 6.93 (d, / = 7,8 Hz,
2H, Ar-H), 7.63-7.73 (m, 7H, Ar-H), 7.39-7.52
(m, 8H, Ar-H), 8.07-8.10 (d, J = 7.8 Hz, 2H,
Ar-H), 8.41 (d, J = 8.4 Hz, 2H, Ar-H).
5 37.06, 61.09, 115.16, 122.36, 124.09, 126.80,
126.87, 127.34, 128.32, 128.37, 130.29, 130.69,
130.70, 130.73, 135.12,135.38, 135.84, 138.57,
138.86, 141.47, 147.33, 150.14, 156.98, 191.28.
598 (M'") 599 (M'"+l).
Calculated for molecular formula C35H26N4O4S.
C, 70.22; H, 4.38; N, 9.36.
C, 70.14; H, 4.35; N, 9.34%.
70
Chapter II, Section 1
3-(4-Nitrophenylmethylt!no)-S[(4~biphenyloxy)methyl]-4~(4-nitrophenyl)-4H‘-ls2,4-"
triazole (99) Yield: 78%; Light yellow crystals, m.p. 160-162 °C, Rf = 0.28 («-
hexane : ethyl acetate; 4:6).
NO2
IR (KBr) cm’*
'H NMR (300 MHz, CDCI3)
13C NMR (1§0 MHz, CBCI3)
; 2912, 1546, 1106.
: 5 4.56 (s, 2H, S-CH2), 5.13 (s, 2H, O-CH2), 6.92
(d, 8.7 Hz, 2H, Ar-H), 7.31 (t, / = 7.2 Hz,
IH), 7.39-7.58 (m, lOH, Ar-H), 8.13 (d, 7 - 8.7
Hz, 2H, Ar-H), 8.37 (d, J = 9.0 Hz, 2H).
: 5 36.86, , 59.78, 114.97, 115.14, 123.25, 126.26,
126,84, 128.74, 129.82, 130.15, 130.26, 130.46,
130.86, 131.24, 134.92, 137,83, 146.53, 146.58,
155.86, 156.
: 539 (M"'), 540 (M" + l).
: Calculated for molecular formula C28H21N5O5S.
; C, 62.33; H, 3.92; N, 12.98.
: C, 62.48; H, 3.89; N, 12.96 %.
2-f5-{(4-Biphenyloxy)methyl}-4-(2-methylphenyl)~4H-l,2,4-triazol-3-ylthio]-l-(4-
bromophenyl)ethanone (100) Yield; 82% ; White Crystals, m.p= 197-199 ®C, Rf =
0.40 («-hexane : ethyl acetate; 4:6)
Maldi-MS {m/z)
Elemental Analysis
Calculated
Found
Br
IR (KBr) cm '
'H NMR (300 MHz, CDCI3)
2916,1534,1106.
6 2.10 (s, 3H, Ar-CHa), 4.90-4.93 (m, 2H, S-
CH2) 5.05 (s, 2H,0 -CH2), 7.90 (d, ./ = 8.4 Hz,
2H), 7,63 (d, J - 7.5Hz, 2H), 7.30-7.50 (m,l OH,
71
Chapter II, Section 1
13C NMR (75 MHz, CDCI3)
Maldi-M S {m/z}
Elemental Analysis
Calculated
Found
Ar-H),7.19 (d, J = 8.4Hz, IH), 6.88 (d, . / - 8.7
Hz, 2H),
: 5 17.32, 38.41, 60.02, 115.05, 126.76, 126.87,
127.41, 128.20, 128.75, 129,35, 130.05, 130.91,
131.39, 131.64, 132.21, 133.94, 134.83, 136.40,
140.50, 152.11, 152,71, 157.11, 192.13.
: 570 (M '), 571 (M'"+l).
; Calculated for molecular fonnula
C3oH24BrN302S.
: C, 63.16; H, 4.24; N, 7.37.
: C, 63.21; H, 4.21; N, 7.40%.
2-[S-{(4-Biphenyloxy)methyl}-4-(2~methylphenyl)-4H-l,2,4-triazol-3-ylthio]-l-(4- methoxyphenyl)ethanone (101) Yield: 80%; White Crystals, m .p. 194-196 “C, Rf ==
0.41 («-hexane : ethyl acetate; 4:6).
-1.IR (KBr) cm
'H NM R (300 MHz, CDCI3)
NM R (75 M Hz, CDCI3)
ES-MS (m/z)
E lem ental Analysis
Calculated
Found
CH,
2926, 1686, 1534, 1107.
5 2.10 (s, 3H, Ar-CHj), 3.87 (s, 3H, O-CH3),
5.01-5.03 (m, 2H, S-CH2), 5.06 (s, 2H, O-CH2),
6,88 (d, J = 8.1 Hz, 2H), 6.95 (d, J - 8.4 Hz,
IH), 7.21 (d, J - 6.3 Hz, 2H), 7.29 (t, J = 6.3
Hz, IH), 7.37-7,50 (m, 9H, Ar-H), 8.04 (d, J =
6.0H z,2H ).
6 17.39, 41,31, 55.55, 59.89, 114.02, 115.02,
126.72, 126.84, 127.42, 127.79, 128.18, 128.71,
130.95, 131.65, 134.86, 136.35, 140.46, 157.02,
164,25, 191.29.
522 (M’"+l), 523 (M^+l).
Calculated for molecular formula C3 jH27N3 0 3 S.
C, 71.38; H, 5.22; N, 8.06.
C, 71.31; H, 5.18; N, 8.04%.
72
Chapter II, Section }
2-[5~{(4~Biphenyloxy)methyl}-4-(2~methylphenyl)-4H~l,2,4"triazol-3-yhhioJ~I~(4~
phenyl)ethanone (102) Yield; 6 8%; White Crystals, m.p. 158-156 "C, Rf = 0.43 («-
hexane ; ethyl acetate; 4;6).
IR (KBr) cm"'
'H NMR (300 MHz, CDCI3)
NMR (100 MMz, CDCI3)
Maldf-MS (m/z)
Elemental Analysis
Calculated
Found
2926, 1520, 1108.
5 2.11 (s, 3H, Ar-CHa), 4.96-5.00 (m, 4H, O-
CH2, S-CH2), 6.89 (d, ./= 8.7 Hz, 2H), 7.21 (d, J
= 7.8 Hz, IH), 7.22 (t, J = 6.9 Hz, IH), 7.32-
7.52 (m, 8H), 7.62 ( d, J - 6 . 9 Hz, 2H,), 7.71 (d,
J= 8 .1 Hz, 2H), 8.11 ( d , J = 8.1 Hz, 2H).
5 16.46, 36.86, 59.89, 114.97, 115.14, 123.17,
126.28, 126.78, 128.72, 129.82, 130.10, 130.22,
130.40, 130.84, 131.20, 137.72, 134.82, 146.20,
146.55, 155.73, 191.62.
491 (M^), 492 (M++1).
Calculated for molecular formula C36H29N3O2S.
C, 73.30; H, 5.13; N, 8.55.
C, 73.34; H, 5.10; N, 8.53%.
2-[S~{(4-Biphenyloxy)methyl}-4-(2-methylphenyl)-4H-I,2,4-triazol-3-ylthio]-l~(4-
phenyl-phenyl)ethanone (103) Yield: 74%; White Crystals, m.p. 168-170 °C, Rf =
0.45 (rt-hexane : ethyl acetate; 4:6).
.-1IR (KBr) cm’
‘H NMR (300 M Hz, CDCI3)
2926, 1524, 1109.
5 2.11 (s, 3H, Ar-CHa), 4.96-5.00 (m, 4H,
O-CH2, S-CH2), 6.89 (d, J = 8.7 Hz, 2 H), 7.21
(d, J = 7.8 Hz, IH), 7.22 (t, J = 6.9 Hz, 2H),
73
Chapter II, Section 1
13C NM R (100 MHz, CDCI3)
M aldi-M S (zw/z)
Elem ental Analysis
Calculated
Found
7.37-7.51 (m, 11H), 7.63 (d, J - 6,9 Hz, 2H),
7.71 (d, J = 8.1 Hz, 2H), 8.11 (d, 8.1 Hz,
2H).
5 16.46, 36.86, 59.89, 114.97, 115.14, 123.17,
126.28, 126.78, 128.72, 129.82, 130.10, 130.22,
130.40, 130.84,131.20,137.72, 134.82, 146.20,
146.55, 155.73, 156.84,162.94,191.62.
567 (M"), 568 (M"'+l).
Calculated for molecular formula C36H29N 3O2S.
C, 76.16; H, 5.15; N, 7.40.
C, 76.22; H, 5.16; N, 7.41%.
5[~{(4-Biphenyloxy)methyl}-3-(ethylthio)-4-o-tolyl]~4H-l,2,4"triazole (104) Yield:
76%; White Crystals, m.p. 174-176 "C, Rf = 0.40 («-hexane : ethyl acetate; 4:6)
. - 1IR (KBr) cm'
NM R (300 MHz, CDCI3)
13C NMR (1 0 0 MHz, CDCI3)
M aldi-M S (m/z)
Elemental Analysis
Calculated
Found
: 2926, 1524, 1109.
: 5 2.08 (s, 3H, Ar-CHa), 2.74 (t, J = 3 .8 Hz, 3H,
S-CHa), 3.29 (q, J = 4.6 Hz, 2 H, S-CH2-), 5.05
(s, 2H, O-CH2), 6.88 (d, 8.4 Hz, 2 H), 7.17
(d, J= 8,4 Hz, IH), 7.25-7.51 (m, I OH).
: 5 15.24, 16.36, 29.12, 59.89, 114.97, 115.14,
123.27, 126.27, 126.89, 128.79, 129.89, 130.12,
130.27, 130.48,130.89,131.27, 134.91, 137.73,
146.23, 146.57,155.72, 155.84,162.92.
401 (M'"), 402 (M'^+1).
Calculated for molecular formula C24H23N3OS:
C, 71.79; H, 5.77;N, 10.47
C, 71.69; H, 5.75; N, 10.46%.
74
Chapter II, Section 1
5[-((4-Eiphenyloxy)methyl}-3-(methylthio)-4-o~tolyl]-4H~l,2,4~triazole (105) Yield:
72%; White Crystals, m.p, 110-112 °C, Rf = 0.41 (M-hexane : ethyl acetate; 4:6)
IR (K B r) cm-' ; 2916, 1534, 1106.
'H NM R (300 MHz, CDCI3) ; 5 2.08 (s, 3H, Ar-CHa), 2.70 (s, 3H, S-CH3), 5.05
is, 2H, O-CH2), 6.88 (d, J - 8.4 Hz, 2H), 7.17
(d ,, / - 8.4 Hz, 1H), 7.25-7.51 (m, 1 OH).
NMR (100 MHz, CDCI3) : 5 14.54, 15.38, 59,89, 114.97, 115.14, 123.27,
126.27, 126.89, 162.97, 128.79, 129.89, 130.12,
130.27, 130.48, 130.89, 131.27, 134.91, 137.73,
146.23, 146,57, 155.76, 156.89.
M aldi-M S (m/z) : 387 (M^), 388 (M^+1).
Elem ental Analysis ; Calculated for molecular formula C23H2iN3 0 S.
Calculated : C, 71.29; H, 5.46; N, 10.84.
Found ; C, 71.62; H, 4.51; N, 10.69%.
2,5.1,2.2. Biological activity
All the novel synthesized compounds were screened for their biological activities.
Albino Wistar rats of either sex (150-200 g) were obtained from Central Animal
House, Hanidard University, New Delhi. Fourteen hours before the start o f the
experiment, the animals were sent to lab and fed only with water ad libitum. The
experiments were performed in accordance with the rules of Institutional Animals
Ethics Committee (registration number 173-CPCSEA). The animals were divided into
groups of six animals each. Bacterial and fungal cultures were obtained from
Microbiology laboratory, Batra Hospital, New Delhi, India and the screening tests
were carried out at the Microbiology laboratory, Majeedia hospital, Jamia Hamdard,
New Delhi.
75
Chapter II, Section 1
2.5.1.2.2.1. Anti-inflammatorv activity
All the newly synthesized compounds were evaluated for their anti-inflammatory
activity [53] against carrageenan-induced acute paw oedema. The first group of rats
was treated with 0.1 w L o f 0.5% CMC suspension orally (control), second group was
administered with a dose of 10 mg/kg of the suspension of Ibuprofen (standard) and
the test group was treated with equimolar dose o f the suspension of test compounds
relative to standard dmg. After 30 min the animals were injected with 0.1 mL o f 1%
caiTageenan in normal saline subcutaneously to the sub-plantar region of right hind
paw. The paw volume was measured at 0 h, 1 h, 3 h and 5 h, by using
plethysmometer. The amount o f oedema in the drug-treated groups was compared in
relation to the control group with the coiTesponding time intervals.
The percent oedema inhibition was calculated from the mean effect in the control and
treated animals according to the following equation.
Percentage inhibition = 100 (1 -- Vt/Vc),
Vt = mean increase in paw volume of test
Vc = mean increase in paw volume of control gi'oup of rats.
The results of anti-inflammatory activity are presented in table 1.2 and figure 1,1.
2.5.1.2.2.2, Analsesic activity
The analgesic activity was detennined using tail flick method [54]. The animals were
weighed and divided into control, standard, and test groups and each group contained
six rats. The first group of rats was treated with 0.1 mL o f 0.5% CMC suspension
orally (control), second group was administered with a dose o f 10 mg/kg of the
suspension of Ibuprofen (standard) and the test group was treated with equimolar dose
o f the suspension of test compounds relati ve to standard drug. After administration of
drug, reaction time was measured at 0 min., 30 min., 60 min., and 120 min., by
immersing the tail in hot water (55 ± 0.5 ‘’C). Analgesic activity was expressed as
reaction time of tail flicking, compounds having analgesic activity will show more
reaction time than the control. The results are presented in table 1.3 and figure 1.2.
2.5.1.2.23. Histovatholoslcal studyFor the histopathological study, rats were sacrificed under light anesthesia after 4 h of
the doses (3 times to the dose used for anti-inflammatory) and their stomach
76
Chapter II, Section 1
specimens were removed and kept into 10 % formalin solution. A longitudinal section
of stomach along the greater curvature, which included the ulcer base and both sides
of the ulcer margin, was taken and fixed in 10% fonnalin for 24 h at 4 "C and
embedded in white solid paraffin. Moiphological examination was performed with
Haematoxylin and eosin staining to analyze histological changes and examined under
microscope. The results are given in table 1.4 and figure 13.
2.5.1.2.2.4. h t vitro Antimicrobial activity
The antimicrobial studies of the synthesized compounds were carried out against
different bacterial and fungal strains. All bacterial and fungal strains were obtained
from the Batra Hospital, New Delhi, India and Department of Biochemistry, Jamia
Hamdard, New Delhi, India as follows
Bacterial Strains
1. Pseudomonas aeruginosa (ATCC 27853)
2. Klebsiella pneumoniae (ATCC 700603)
3. Escherichia coli (ATCC 25922)
4. Staphylococcus aureus (ATCC 25923)
Fungal Strains
1. Candida albicans (ATCC 10231)
2. Candida krusie (ATCC 6258)
3. Aspergilus niger (MTCC8189)
4. Aspergilus flavus (MTCC 277)
All the newly synthesized compounds were dissolved in dimethyl sulfoxide (DMSO)
to prepare chemicals of stock solution of 2 mg/mL.
2.5.1.2.2.4.I. A^ar-well diffusion method
Simple susceptibility screening test using agar-well diffusion method [55] was used.
Each microorganism was suspended in Mueller Hinton (MH) (Difco, Detroit, MI)
broth and diluted approximately to 10'’ colony foi-ming unit (cfu)/mL. They were
“ flood-inoculated” onto the surface of MH agar and Sabouraud Dextrose Agar
(SDA) and then dried. For Candida albicans, Candida krusie, Aspergilus niger and
Aspergilus flavus, SDA were used. For Pseudomonas aeruginosa, Klebsiella
77
Chapter II, Section I
pneumoniae, Macconcy agar was used and for Escherichia coli and Staphylococcus
aureus Muller Hinton agar were used. Six-millimeter diameter wells were cut from
the agar using a sterile cork-borer and 2 00 ng and 100 iig o f the test compounds were
delivered into the wells. The plates were incubated for 18 h at 35 "C for bacteria and
48 h for fungal strains. Antimicrobial activity was evaluated by measuring the zone of
inhibition against the test organism. Ampicillin (200, 100 ^ig) and Fluconazole (200,
100 (ag) were used as standard drugs. Dimethyl sulfoxide was used as solvent
(controls). The antimicrobial activity results are summarized in table l.S.
78
Chapter II, Section I
2.5oL3. ConclusfoeIn conclusion a focused library of biphenyl based 1,2,4-triazoie derivatives has been
successfully synthesized and evaluated for their anti-inflammatox'y, analgesic and
antimicrobial activities. From the results, it is concluded that seven compounds 68 ,
74, 8 8 , 96, 97, 99, and 101, out of thirty-nine compounds showed good anti
inflammatory activity against carrageenan induced rat paw oedema method.
Compounds 68 showed better anti-inflammatory activity (79.84, 83.03%) than
standard drug Ibuprofen (78.89 and 80.10%) at 3 h and 5 h respectively. Compounds
99 and 101 showed better analgesic activities (3.81, 3.96 and 3,08) and (3.88, 3.98
and 2.95 sec) respectively compared to the standard drug Ibuprofen (3.71, 3.95 and
2.76 sec) at 30, 60 and 120 min. All the tested compounds showed less ulceration than
standard drug Ibuprofen whereas compound 74 did not caused any ulceration. The
antimicrobial activity revealed that most of the compounds showed moderate to
significant activity. Compounds having nitro, chloro, bromo and fluoro group showed
better activity.
79
.i.ert ™1 &3 »i 83
OSB,i\ S7
m
■O'
h
>5Sg^
Hr‘?3 ■ iS ,
f|H " | j
r i' . i
*? 4 s *fj ” Hr* % 6S.W KM |.3«
f .; i p i ; ,- 'ji :,!..
rI f
'H NmR spectrum o f compound 68
§iSSSSSSSSS
i '3*'
,r..v • .V '•' : i I '• 4 m i • r; . -'s ? -4 I
■U ■ ... . v . ■ ■ «■ : ■''.■■■ . ' ■ ■■ ■ . .■
; *'' (. "’ r ' ,«£ J £«.. « I-,., 5 k l
r a “ .« 'c i | I."’ ^ - r - - ^ ‘I ^f I * I F i ^i i r i i
' C NMR spectrum of compound 68
Oo
.is.oo
Oioo
CO
8 ooCf>oo
w050501
k)<7>
-O'4
N3.. CO
K>o
, . pw>05
p)K>01
01M4h.cn
Mass spectrum of compound 68
■o
"H» Wma)
f i gi'll i - I «2 i
5
P
TransRittaftoe
26’£«S-<
o ifmm
.......1
Les'SOTi.-
ere€#i*™f fS S S I-- “-*»««= s= = r:
s 's r i ,- " -
m m z i ..
. >w '« re i-
oN3
ftzetj»~««essi£::: 6frir‘#9£.' " ‘■«*“ssssss;2J
§S9‘a®^ ' ■
'"H:ssn
..." : : : j
IR spectrum of compound 68
Chapter II
Section 2
Synthesis of biphenyl based hydrazones of 1,2,4-
triazole derivatives and their Biological Activities
Chapter II, Section 2
2 .4JJ , Introduction
As discussed in introductory part o f this chapter, hydrazone, triazole and biphenyl
derivatives exhibit different biological activities. In view of the biological importance
of hydrazones, 1 ,2,4-triazoles and biphenyl systems, a focused library o f conjugates
have been synthesized and evaluated for their anti-inflammatory, analgesic and
ulcerogenic studies. The synthetic route is given below.
SCHEM E II
o- T \N -SH
63,65
o-N"NH
Abs. alcohol Na Metalethylbromoacetate reflux 4-6 hr.
v\ / - \ \ o
Abs. alcohol NH2NH2.H2O reflux, 4-6 hr.
Abs. alcohoi Aromatic aldehyde reflux, 4-6h.
V\ //> ^ \
106,107
N "N
N
110-123
HN-NH2
o
108, 109
N-N7 >
N
H N -N
O
80
Chapter 11, Section 2
Reaction of triazoles (63, 65) with ethylbromoacetate in presence o f sodium metal in
absolute alcohol afforded ethyl esters (106) and (107), which readily yielded
hydrazides (108) and (109) by refluxing with hydrazine monohydrate in absolute
alcohol. The hydrazides when further reacted with different aromatic aldehydes in
absolute alcohol yielded hydrazones (110-123). All the synthesized compounds have
been completely characterized on the basis o f their detailed spectral data and all the
novel compounds of this series were evaluated for their biological activities.
The physical data o f all the novel synthesized compounds are given in table 2.1.
Table 2.1 Physical data of biphenyl based hydrazones o f 1,2,4-triazole derivatives
Comp. no. S tr u c tu r e % Y ield m.p. ( “C )
110
01
68 146-148
111
0Cl
72 187-189
112
< ) °C(
71 168 -170
113
< )
Cl
68 144 -146
114
< ) ?Cl * 3
78 173-175
115
9 ^Cl CHg
72 171-173
81
Chapter II, Section 2
1 1 6 H HO ^ N-N >=\ .CH ,
O 0 - C H 3
Cl
6 6 1 9 3 - 1 9 5
1 1 7 69 1 7 9 - 1 8 1
1 1 8
<) «
Cl
6 8 1 7 6 - 1 7 8
1 1 9
QCl
7 2 1 8 4 - 1 8 6
1 2 0
< )Cl
6 8 1 8 1 - 1 8 3
1 2 1
O O - C H j
Cl
15 1 8 0 - 1 8 2
1 2 2
0O -C H g
8 0 1 6 8 - 1 7 0
1 2 3
0° ~ C H 3
7 8 1 6 5 - 1 6 7
82
Chapter II, Section 2
2.4.2.1, Results and discussion2.4.2.1.1, Analytical
A focused library of fourteen compounds 110-123 were synthesized starting from 3-
mercapto-l,2,4-triazole as outlined in scheme-II. S~alkylation of 3-mercapto-l,2,4-
triazole 63 and 65 to 106 and 107 were confirmed from the 'H NMR spectmm. The
appearance of two signals at 6 1.25 and 1.26 each (t, J ~ 7.2 Hz, 3H, -CH3) and at 6
4.20 (q, J — 7.2 Hz, 2H, 0 -CH2) integration for tlve protons suggested the presence o f
ethoxy group. The presence of strong absoiption band at 1673-1675 cm '' and absence
of bands for S-H, at 2680-2690 cm’’ confirms the S-alkylation of thiol group. The
compounds 106 and 107 readily yielded their hydrazides 108 and 109 by their
reaction with hydrazine monohydrate. The formation of these hydrazides were
confirmed from the appearance of signals at 6 8.91 (s, IH, N-FI) and 5 1.62 (s, 2H,
NH2) and disappearance of ethyl group signals. These hydrazides were then converted
into the target molecules 110-123 which was confirmed from IR (absorption bands for
N=C, 1655-1687 cm"'), extra signals in aromatic region o f 'h NMR spectmm due to
introduction of phenyl ring of aldehydes to the target molecules (hydrazones) and
finally confinned from their mass spectra.
All the newly synthesized compounds 110-123 were screened for their biological
activities (anti-inflammatory, analgesic and ulcerogenic activities). These novel
hydrazones showed therapeutic efficacy.
2.4.2.1.2, Biological activity
2,4.2.1.2.1. Anti-inflammatorv activity
The results o f anti-inflammatory activity are summarized in table 2.2 and figure 2,1.
Among the tested compounds, two compounds 115 and 120 were found significantly
active (83,76, 75.64 % and 82.84, 81.13 % inhibition) when compared to the
standard drug Ibuprofen (78.89 and 80.10 % ) in paw oedema at 3 h and 5 h
respectively whereas compounds 116 and 121 showed moderate activity. All the data
were analyzed by one-way ANOVA test followed by Dunnett’s test in carrageenan
induced rat paw oedema model in rats.
83
Chapter 11, Section 2
Table 2.2. Anti-inflammatory activity of biphenyl based hydrazones of 1,2,4-triazole derivatives
Compounds Change in paw oedema volume (mL) After drug treatment
Anti-inflammatory activity
% Inliibition
3h 5h 3h 51i110 0.35±0.022’ 0.32±0.033' 43.18 45.11111 0.40±0.044"“ 0.45±0.022“ 35.00 42.49112 0.48±0.022"’‘ 0.45±0.022"* 22.07 22.81113 0.40±0.039“ 0.43±0.042"^ 35.06 26.24
114 0.35±0.022' 0.41±0.030“ 43.18 29.67
115 0.10±0.036"' 0.10±0.025"' 83.76 82.84116 0.33±0.033" 0.18±0.030'” 46.42 69.12117 0.35±0.042' 0.25±0.021'' 43.18 57.11
118 0.46±0.036“ 0.43±0.033“ 25.32 26.24119 0.40±0.036'‘® 0.43±0.021"" 35.06 26.24
120 0.15±0.022” ' 0.11±0.026'” 75.64 81.13121 0.25±0.036'" 0.18±0.030"’ 59.41 69.12122 0.33±0.05' 0.30±0.033' 46.42 48.54123 0.36±0.033' 0.30±0.036' 41.55 48.54
Ibuprofen 0.13±0.033'" O.lliO.OH'" 78.89 80.10Control 0.615±0.047 0.582±0.030 — —
Data analyzed by one way ANOVA followed by Dunnett’s Y test (n=6), *p<0.05, **p<0.01 & **‘P<0.001 significantly different from standard; ns, not significant
Antiinflammatory Activity
■ 3 hr fcJShr
Figure 2.1. Anti-inflammatory activity of biphenyl based hydrazones of 1,2,4- triazole derivatives
84
Chapter 11, Section 2
2.4.2.1.2.2. A n a ls e s ic a c t iv ity
The results of analgesic activity are summarized in table 2.3 and figure 2.2. The
tested compounds 115 and 120 showed comparable analgesic activity (reaction time)
with the standard drug Ibuprofen. All data were analyzed by one-way ANOVA test
followed by Dunnett’s test.
Table 2.3. Analgesic activity o f biphenyl based hydrazones o f 1,2,4 triazole
derivativesCompounds Tail flick latency in second.
0 min 30 min. 60 min. 120 min
115 I.93±0.136 3.86±0-346’ 3.88±0.243’ 2.85±0.237“
120 2.08±0.320 3.8(W).233' 3.22±0.264' 2.94i0.232 “
121 2.03±0.320 2.64±0.136“ 2.84±0.190”* 2.96±0.230“
Ibuprofen 1.98±0.307 3.71±0.388* 3.95±0.470* 2.76±0.212"*
Control 2.03±0.330 1.94±0.236 2.02±0.192 2.76±0.212
Data analyzed by one way ANOVA followed by Dunnett’s 'V test (n=6), *p<0.05, **p<0.01 & ***p<0.001 significantly different from standard; ns, not significant
Analgesic activity
xOiiiiii * 3 0 mill ‘<60iiiin n i2 0 iii ii i
5
4.5
4
1V 3 EV 2.5 cI 2i 1 . 5
1
0.5
0
115 120 121 ibiipiofmi
Compounds
Figure 2.2. Analgesic activity of biphenyl based hydrazones of 1,2,4-triazole derivatives
85
Chapter II, Section 2
2,4.2,1.2.3, Histooatholoeical Study
When compared with Ibuprofen, compound 115 did not cause any gastric ulceration
and disruption of gastric epithelial cells at the given doses. Hence gastric tolerance to
this com.pouud was better than, that of Ibuprofew indicating that carbox,ylic group
pi-esent in the Ibuprofen is responsible for ulceration, Bhandari et al. [52]. Stomach
wall of Ibuprofen treated group at low power (lOx) photomicrograph showed damage
of the mucosa and the sub mucosa. Stomach wall o f the same section at high power
(40x) photomicrogi'aph showed desquamated epithelial cells in the lumen whereas
tested compounds 115, 120 and 121 treated animals showed significant surface
epithelial damage and slightly submucosal damage however there was lesser damage
in comparison to the Ibuprofen, Stomach wall o f compound 115 treated animal
showed no damage of any layer. The results are shown in table 2.4 and figure 2.3.
Table 2.4. Haematoxylin and Eosin immunohistochemical staining of gastric ulcers
after ulcer induction in rats
Compounds Surface Epithelial Damage
SubmucosalDamage
DeepMucosalDamage
MuscularLayer
Damage
115 “ - - -
120 ++ + - -
121 ++ + - -
Control _ - -
Ibuprofen +++ +++ - -
(+; ++; +++) Increase ulceration; {-) no ulceration
Continued,
86
Chapter II, Section 2
. WVI f l .* .
5
* - r ‘- •--^
115 (lOX)
I I
120 (lOX)
«5r s / i
115 (40X)
120 (40X)
121 (lOX) 121 (40X)
C on tin u e d .
87
Chapter II, Section 2
Ib u p i ofnt <10X> Ibnpi ofm (40X)
L U M E N L U M E N
£ \ * -t h M y 'I -V Mg -I < 1 HR*
ronti ol <40\')
Figure 2.3 Haematoxylin and Eosin immunohistochemical staining of gastric ulcers after ulcer induction in rats.
88
Chapter 11, Section 2
IA.2.2. Expeiim entel2.4.2.2.L Chemistry
All chemicals (reagent gi'ade) used were commercially available. Melting points were
measured on a VEEGO-VMP-DS melting point apparatus and are uncorrected. ’H
NMR was recorded on a Bruker DPX 400, 300 instruments respectively in CDCI3/
DMSO-df, using TMS as internal standard. 'H NMR chemical shifts (6) and coupling
constants ( J ) are given in ppm and Hz respectively. Mass spectra were recorded on a
Jeol JMS-D 300 instrument fitted with a JMS 2000 data system and operated at 70 eV
and Maldi-MS (AB-4800). Mass-spectrometric (MS) data are reported in m/z.
Elemental analysis was earned out using Elementar Vario EL III elemental analyzer.
Elemental analysis data are reported in % standard.
General Procedure fo r synthesis o f ethyl ester o f 3-mercapto-l,2,4-triazole (106,
107)
To a solution of 3-mercapto-l,2,4-triazole (61, 63, 10 mmol each) in absolute alcohol
(50 mL) was added sodium metal and ethyl bromoacetate (equimolar) and refluxed
for 4-6 hr. The resulting solution was concentrated under reduced pressure and poured
on crushed ice; the precipitate so obtained was filtered, washed with cold water, dried
and recrystallized from alcohol.
Ethyl2-[5((4-biphenyloxy)methyl}-4-(4-methoxyphenyl)-4H-l,2,4-triazol-3-ylthio]
acetate (106) Yield: 78%; White Crystals, m.p. 138-140 ""C; Rf - 0.56 (CHCI3:
M eOH;9;l).
IR (KBr) cm"’ ; 3062, 2989, 1673, 1611, 1241, 1093.
'H NMR (300 MHz, CDCI3) : 5 1.26 (t, J = 7.2 Hz, 3H, CH2-CH 3), 3.85 (s,
3H, O-CH3), 4.08 (s, 2H, S-CH2), 4.20 (q, ,/ =
7.2 Hz, 2H, O-CH2), 5,09 (s, 2H, O-CH2), 6.95-
89
Chapter II, Section 2
Maldi-MS (m/?:)
Elemental Attaiysis
Calculated
Found
7.01 (m, 4H), 7.26-7.31 (m, 3H ), 7.40 (t, 7.5
Hz, 2H), 7.50-7.53 (m, 4H, Ar-H).
475 (M"-), 476 (M'"+l)
Calculated for molecular formula C26H25N 5O4S;
C, 65.67; H, 5.30; N, 8.84
C, 65.57; H, 5.29; N 8.83%
Ethyl-2~[5 {(4-biph enyloxy) m ethyl}-4~(4-ch lorophenyl)-4H-l, 2,4-triazol-3-
ylthio]acetate (107) Yield: 72% ; White Crystals, m.p. 128-130“C; Rf
(CHCI3: MeOH; 9:1).
- 0.55
Q N
IR (KBr) cm-’
*H NM R (300 MHz, CDCI3)
Maldl-MS (w /4
Elemental Analysis
Calculated
Found
: 3058,3025, 1675, 1603, 1241, 1107.
: 5 1.26 (t, 7.2 Hz, 3H, CH2-CH3), 4.09 (s, 2H,
S-CH2), 4.20 (q, J = 7.2 Hz, 2H), 5.09 (s, 2H, O-
CH2), 6.94 (d, J = 8.7 Hz, 2H), 7.25-7.43 (m,
5H ), 7.46-7.52 (m, 6H, Ar-H).
480 (M^), 482 (M'"+2)
Calculated for molecular formula C26H25NSO4S:
C, 65.67; H, 5.30; N, 8.84
C, 65.57; H, 5.29; N 8.83%
General procedure fo r synthesis o f hydrazide (108, 109)
The ethanolic solution of compounds 106, 107 (10 mmol) and hydrazine monohydrate
(10 mmol) was refluxed for 4-5 h. After the completion o f reaction, the reaction
mixture was cooled, poured on crashed ice, the precipitate so obtained was filtered,
washed with cold water, dried and recrystallized from alcohol.
90
Chapter 11, Section 2
2-[5 {(4-hiph enyioxy) m ethyl}-
acetohydrazide (108) Yield
(CHCI3 : MeOH; 9:1).
'4-(4-methoxyphenyi)-4M-l,2,4-triazol-3-yithioJ
:78%; White Crystals, m.p. 241-143 °C; Rf = 0.51
- IIR (KBr) cm"
*H NMR (300 MHz, CDCI3)
MaMWMS (m/z)
Elemental Analysis
Calculated
Found
°\ J '-N \ | iN HN-NH2
o
QC H 3
: 3503, 1683, 1603, 1507.
: 5 1.62 (brs, 2H, -NH2), 3.85 (s, 3H, O-CH3 ), 4.08
(s, 2 H, S-CH2), 5.07 (s, 2 H, O-CH2), 6.95-7.01
(m, 4H), 7.25-7.32 (m, 3H), 7.40 (t, J = 7.5 Hz,
2H), 7.46-7.52 (m, 4H, Ar-H), 8.91 (s, IH, N-
H).
: 461 (M”"), 462 (M-'+l)
: Calculated for molecular formula C24H23N5 O3S
: C, 62.46; H, 5.02; N, 15.17
: C 62.39; H 5.01; N 15.15%
2-[5{(4-biphenyloxy)methyl}~4-(4-methoxyphenyl)-4H-l,2,4-triazol-3"ylthio]
acetohydrazide (109) Y ie ld ; 8 8 %; White crystals, m.p, 137-139 °C; Rf = 0.53
(CHCb; MeOH; 9:1).
'— \ A ^-NHa
y / /Cl
IR (KBr) cm-
'H NM R (300 MHz, CDCI3)
Maldi-MS {m/z)
3246, 3058, 2690, 1668, 1241, 1107.
6 2.95 (brs, 2H, NH-NHj), 3.94 (s, 2 H,S-CH2),
5.11 (s, 2H, O-CH2), 6.96 (d, 8.4 Hz, 2H),
7.30-7.32 (m, 3H), 7.43 (t, / = 7.8 Hz, 2H),7.49-
7.54 (m, 6 H, Ar-H), 8.14 (s, IH, N-H).
465 (M""), 467 (M^+2)
91
Chapter II, Section 2
Elemeifttal Analysis
Calculated
Found
Calculated for molecular fomiula C23H20CIN5O2S
C, 59.29; H, 4.33; N, 15.03
C 59.21; H 4.31; N 15.05%.
General procedure fo r synthesis o f hydrazones o f 1,2,4-triazole (110-123)
To a solution of liydrazide 108, 109 (Immol) in absolute alcohol (50 mL) was added
different aromatic aldehydes (equimolar) with few drops o f glacial acetic acid and the
reaction mixture was refluxed for 4-6 hr. After the completion of reaction monitored
by TLC, the reaction mixture was cooled to RT and concentrated under reduced
pressure, the concentrated solution was poured on crushed ice, and the precipitate so
obtained was filtered off and washed with cold water, dried and crystallized in ethanol
to yield the pure hydrazone 110-123.
N'-(3~Methylbenzylidene)-2-f5-{(4-phenylphenoxy)methyl}~4-(4-chlorophenyl)-4H~
l,2,4-triazol-3«yithio]acetohydrazide (110) Y ield’. 6 8 %; White crystals, m .p .146-148
“C; R,- = 0.50 (CHCI3 : MeOH; 9:1).
IR (KBr) cm"
’H NM R (300 MHz, CDCI3)
NM R (75 MHz, CDCia)
Maidl-MS im/z)
Elemental Analysis
^ V _ / \N
; 3254, 3058, 2912, 1608, 1241, 1107.
: 8 2.29 (s, 3H, Ar-CHs), 3.96 (s, 2H, S-CH2),
5.04 (s, 2H, O-CH2), 6 . 8 6 (d, / = 7.8 Hz, 2H),
7.15 (d, J=1.S Hz, 2H), 7.22 (t, 7 = 7.2 Hz, IH),
7.33 ( t , J = 7.2 Hz, 2H), 7.43-7.40 (m, 9 H), 9,41
(s, IH, N-H), 7.56 (d, 2H, J= 7.5 Hz).
: 5 21.47, 39,35, 59.12, 115.01, 115.05, 134.92,
135.23, 136.38, 136.95, 140.28, 140.41, 140.73,
140.83, 145. 52, 148.98, 154.10, 156.67, 156.85,
164.48, 169.14.
: 568 (M‘"), 569 (M’ +l)
: Calculated for molecular fomiula C31H26
CIN5O2S
92
Chapter II, Section 2
Calculated
Found
: C, 65.54; H, 4.61; N, 12.33
: C 65.57; H 4.62; N 12.34%
N'-(4-Chlorobenzylidene)-2-[5-((4-phenylphenox}>)methyl}-4-(4-chlorophenyl)-4H-
l,2,4~triazol-3-ylthio]acetohydrazide (111) Yield: 68%; White crystals; m.p.187-189
"C; R f - 0.53 (CHCb; MeOH; 9:1).
IM (KBr) cm '‘
NM R (300 MHz, CDCI3)
’•'C N M R (75 MHz, CDCI3)
Maldi-MS ( m/z)
Elemental Analysts
Calculated
Found
: 3089,2977, 1683, 1608, 1491, 1396, 1233,
1092,835,763.
; 5 4.09 (s, 2H, S-CHa), 5.06 (s, 2H, O-CH2),
6.87(d, 5.7 Hz, 2H), 7 .2 1 -7 .3 5 (m, 6H), 7.41-
7.42 (m, 8H), 8.06 (d, J = 6.6 Hz, IH), 8.26 (s,
IH, N-H), 8.52 (s, IH).
; 5 39.16, 59.64, 114.70, 126.25, 126.47, 127.77,
128.09, 129.80, 127.93, 128.34, 128.46, 129.61,
130.79, 131.89, 132.13, 134.37, 135.85, 139.91,
143.09, 146.76, 151.74, 151.39, 156.36, 156.48,
164.05, 168.59.
: 587 (M'"), 588 (M^+1)
; Calculated for molecular fomiula
C30H23N3CI2N5O2S
:C , 61.23; H, 3.94; N, 11.90
;C 61.25; H 3.95; N 11.92%
93
Chapter II, Section 2
N^-(3-Chloroben7.ylidene)-2-[S-{(4~phenylphenoxy)methyl}-4-(4-chlorophenyl)-4H-
1,2,4- triazolS-ylthioJacetohydrazide (112) Yield; 71%; White flakes; m,p.l68-170
"C; Rf=0.54(CHCb: MeOH; 9:1).
.-IIR (KBr) cm"
NMR (300 MHz, CDCI3)
13C NMR (100 MHz, CDCI3)
Maldi-MS {m/z)
Elemental Analysis
Calculated
Found
: 3066, 2940, 1684, 1605, 1491, 1234, 1093.
: 5 4.02 (s, 2H, S-CH2), 5.06 (s, 2H, O-CH2),
6 .8 6 (d, J = 7.2 Hz, 2H), 7.25-7.37 (m, 7H),
7.41-7.47 (m, 7H), 7.63 (d, J - 7.2 Hz, 2H),
8.11 (s, IH, N-H).
: 5 34.28, 59,74, 115.10, 126,79, 127.00, 127,18,
128.04, 128.26, 128.29, 128.35, 128.48, 128.75,
129.66, 130.39, 131.28, 131.36, 134.42, 135.00,
135.26, 136.54, 137.00, 140.37, 140.49, 141.97,
145.12, 156.72,156.86, 164.80, 169.28.
: 588 (M'"), 589 (M-^+1).
: Calculated for molecular formula
C30H23 CI2N5O2S.
: C, 61.23; H, 3.94; N, 11.90
: C 61.26; H 3.95; N 11.89%.
N'-(2-Chlorobenzylidene)-2-[5-{(4~phenylphenoxy)methyl}-4-(4-chlorophenyl)~4H~
1,2,4-triazol~3-yltliioJacetohydrazide (113) Yield: 6 8 %; White crystals; m .p.144-146
”C; Rf = 0.52 ( C H C I3 : MeOH; 9:1).
94
Chapter II, Section 2
IR (KBr) cm"*
‘H NM R (3§(S M Hz, CDCI3)
NM R (100 MHz, CDCI3)
Maldi-MS {m/z)
Elemental Analysis
Calculated
Found
3204,3066, 2940, 1684, 1605, 1491, 1233, 1093.
6 4.09 (s, 2H, S-CH2), 5.06 (s, 2H, O-CH2), 6.87
(d, J = 5.7 Hz, 2H), 7.21-7.35 (m, 7H), 7.4-
7.42 (m, 7H), 8.52 (s, IH), 8.26 (s, IH, N-H),
8.06 ( d ,y = 6.6 Hz, IH).
8 34.28, 59.74, 115.04, 115.12, 126.79, 126.99,
127.16, 128.03, 128.24, 128.29, 128.35, 128.46,
128.77, 129.68, 130.37, 131.26, 131.36, 134.41,
135.00, 135.25, 136.50, 137.00, 140.35, 140.47,
141.97, 145.24, 156.72, 156.89, 164.82, 169.20.
587 (M''), 588 (M'^+1).
Calculated for molecular formula
C30H23N3CI2N5O2S
C, 61.23; H, 3.94; N, 11.90
C 61.25; H 3.95; N 11.92%.
N'~(4~methoxybenzylidene)-2-[5-{(4-phenylphenoxy)methyl}-4~(4~chlorophenyl)-
4H-l,2,4-triazol~3-ylthiJacetohydrazide (114) Yield: 78 %; white crystals; ra.p. 173-
175"C; Rf = 0.28 (CHCI3; MeOH; 9;1).
Q
HN ,
N 'O C H ,
O H
IR (KBr) cm '
NMR (300 MHz, CDCI3)
3216,3066, 1675, 1603, 1515, 1492, 1398, 1276,
1097,1005,835,764.
5 3.84 (s, 3H, O-CH3), 4.08 (s, 2H, S-CH2), 5.13
(s, 2H, O-CH2), 6.89 (d, / = 8.4 Hz, 2H), 6.96
(d, J - 8.4 Hz, 2H), 7.31-7.52 (m, 5H), 7.50-
7.55 (m, 6 H, Ar-H), 7.86 (d, / = 8.4 Hz, 2H),
8.14 (s, lH , N-H), 11.52 (s, IH, OH).
95
Chapter 11, Section 2
NMR (100 M Hz, CDCI3)
Maldi-MS (w/s)
Elemental Analysis
Calculated
Found
: 5 35.01, 55.37, 59.28, 114.51, 114.67, 122.64,
124.91, 125.21, 126.28,127,50, 127.95, 128.16,
129.55, 130.66, 134.11, 135.56, 139.69, 144.69,
\47.26, 147.33, 148.21, 148.63, 151.10, 152.54,
156.21, 156.31, 163.37, 168.02.
; 584 (M'X 585 (M^'+l)
: Calculated for molecular formula
C31H26CIN5O3S
;C , 63.75; H, 4.49; N, 11.49
:C , 63.78; H, 4.50; N, 11.51%.
N ’-(3,4-dim eth oxybenzylidene) -2-[S-{(4-phenylph en oxy) m ethyl}-4'-(4-ch lorophenyl)
~4H-l,2,4-triazol~3-ylthio]acetohydrazide (115) Yield: 68%; Wliite crystals;
m.p.183-185 ”C; Rf ^ 0.46 (CHCI3; MeOH; 9:1) .
IR (KBr) cm’’
’H NM R (300 MHz, CDCI3)
'^C NM R (100 MHz, CDCI3)
Maldi-MS {m/i)
Elemental Analysis
; 3216, 3054, 2936, 1684, 1603, 1515, 1492, 1377,
1268, 1134, 1093, 1024, 835, 764, 699.
: 5 3.91-3.99 (m, 6H, 2 O-CH3), 4.15 (s, 2H, S-
CH2), 5.14 (s, 2H, O-CH2), 6.82-7.30 (m, 6H),
7.33-7.62 (m, IIH ), 8.13 (s, IH, N-H).
: 5 34.17, 55.91, 59.89, 108.46, 110.47, 115.05,
123,01, 126.46, 127.02, 128.28,128.36, 128.48,
130.10, 130.39, 135.28, 137.04, 140.29, 145.59,
149.14, 151.92, 152.23, 152.72, 154.16, 156.86,
164.51, 169.01.
:614 (M'"), 616(M'"+l).
: Calculated for molecular formula
C32H28 CIN5O4S.
96
Chapter II, Section 2
Calculated
Found
; C, 62.58; H, 4.60; N, 11.40
: C 62.56; H 4.61; N, 11.38%.
N'-(3)S~di-methoxy-4~hydmxybenzylidene)~2-[S~{(4-phenylpheni>xy)methyl}~4-(4-
chlorophenyl)-4H~l,2,4-triazol-3-ylthio]acetohydrazide (116) Yield: 62%; White
crystals; m ,p.l90-192 "C; Rf = 0.40 (n-liexane ; ethyl acetate; 4:6).
.-IIR (KBr) cm'
'H NM R (300 MHz, CDCI3)
‘'"’C NMR (75 MHz, CDCI3)
M aldi-MS ( w/ j:)
Elemental Analysis
Calculated
Found
; 3180, 3054, 2932, 1662, 1592, 1519, 1494, 1378,
1129, 1005, 836, 760.
: 5 3.89-3.84 (s, 0 -CH3. 6 H), 3.90 (s, 2H, S-CH2),
5.09 (s, 2H, O-CH2), 6.89 (s, IH), 7.03-
6.95 (m, 5H), 7.31 -7.27 (m, 4H), 7.40 (t, J =
7.2 Hz, 2H), 7.51-7.48 (m, 4H), 8.07 (s, IH, N-
H), 11.51 (s, IH, OH).
: 5 38.56, 55.71, 59.15, 114.44, 124.10, 125.93,
126.20, 127.43, 127.78, 127.90, 128.09, 129.29,
129.45, 134.25, 135.51, 139.65, 144.77, 147.21,
152.29,156.23,163.09,167.90.
: 630 (M‘ ), 632 (M‘"+2)
: Calculated for molecular formula
CjiHaaClNsOsS.
: C, 61.00; H, 4.48; N, 11.11
: C 61.04; H 4.47; N 11.12%.
97
Chapter II, Section 2
N'-(3'-ethoxy,4~hydroxybenzylidene)~2-[S-{(4~phenylphenoxy)methyl}-4-(4~
chlomphenyl)- 4H-lf2,4-triazol~3~ylthioJacetohydrazide (117) Yield; 68%; White
crystals; m.p.174-176 "C; Rf = 0.53 (CHCI3: MeOH; 9:1).
-1IR (KBr) cn f
‘H NM R (300 M H z, CDClj)
NM R (100 MHz, CDCI3)
Maldl-MS (w/i:)
Elemental Analysis
Calculated
Found
OH
; 3204, 3066, 1678, 1603, 1492, 1378, 1276, 1089,
834, 764.
: 5 1.31 (t, J = 6.9 Hz, 3 H,-CH3), 3.93 (q, 6.8
Hz, 2H, O-CH2), 4.05 (s, 2H, S-CH2), 5.14 (s,
2 H, O-CH2), 6.80-6.88 (ra, 4H), 6.94 (d, J = 7.8
Hz, IH), 7.23 (d, J = 7.2 Hz, 2H), 7.31-7.43 (m,
lOH), 8.00 (s, IH, N-H), 11.37(s, IH, OH).
: 5 14.61, 35.33, 59.67, 64.26, 114.91, 121.60,
125.55, 126.36, 126.66, 127.86, 128.29, 128.38,
128.55, 129.73, 129.85, 131.07, 134.38, 135.83,
139.97, 144.99, 146.83, 146.92, 148.84, 149.21,
151.47, 152.53, 156.64, 156.70, 163.51, 168.00.
: 614(M'"), 615 (M^+1).
: Calculated for molecular formula
C 3 2 H 2 8 C IN 5 O 4 S .
:C , 62.58; H, 4.60; N, 11.40
; C 62.59; H 4.61; N 11.41%
(E)~N'-(lH-indol~3-yl)methylene-2-f5-{(4-phenylphenoxy)methyl}-4~(4-
chlorophenyl)-4H-l,2,4-triazol-3-ylthioJacetohydrazide (118) Yield: 68%; light
yellow flakes, m.p. 176-178 "C; R, = 0.37 ( C H C I 3 : MeOH; 9:1).
NO ^ //
98
Chapter II, Section 2
IR (KBr) cm-’
'H NM R (300 M Hz, CDCI3)
NM R (100 MHz, CDCI3)
Maldi-MS (?«A)
Elemental Analysis
Calculated
Found
: 3474, 1671, 1613, 1248, 1092.
, 5 4.54 (s, 2H, S-CHa), 4.98 (s, 2H, O-CH2), 6.82
(d, y = 6 Hz, 2H), 7.20-7.05 (m, 3H), 7.40-7.29
(m, 14H), 8.00 (s, IH, N-H), 9.88 (IH , s, OH).
5 35,30, 59.90, 111.28, 111.93, 115.28, 120.61,
121.59, 122.67, 124.03, 126.26, 126.86, 127.75,
128.85, 129.08, 129.83, 130.66, 131.61, 133.57,
134.83, 137.11, 139.61, 141.45, 144.38, 151.65,
151.98, 156.91, 162.26,167.38.
593 (M" ), 595 (M'^+2).
Calculated for molecular fomiula
C32H25CIN6O2S.
C, 64.80; H, 4.25;N , 14.17
C 64.83; H 4.26; N 14.19%.
N ’-(S-nitrobemylidene)~2-[S-{(4-phenylphenoxy)methyl}-4-(4-chlorophenyl)-4H-
l,2,4-tnazol-3-ylthio]acetohydrazide (119) Yield: 68%; White crystals, m.p.184-186
"C; R f=0.31 (CHCI3; MeOH; 9:1).
IR (KBr) cMi'
'h NM R (300 MHz, C D C I 3 )
NMR (100 MHz, CDCI3)
H
; 3305, 1678, 1607, 1516, 1450, 1239, 1092.
: 6 4.06 (s, 2H, S-CH2), 5.14 (s, 2H, O-CH2), 6.96
(d, J = 8.4 Hz, 2H), 7.32-7.56 (m, IIH ), 8.01-
8.08 (m, 2H), 8.14 (d, J== 6.9 Hz, IH), 8.19 (t, J
== 7.5 Hz, IH), 8.31 (s, IH, N-H), 8.49 (s, IH,
Ar-H).
: 5 34.94, 59.47, 114.71, 121.50, 123.80, 126.25,
126.52, 127.79, 127,94, 128.11, 128.35, 129.38,
129.64, 129.82,130.80, 132,04, 134.44, 135.50,
99
Chapter II, Section 2
Maldi-MS {m/z)
Elemental Analysis
Calculated
Found
135.36, 135.97, 139.85, 141.64, 148.08, 151.37,
152.21, 156.49, 167.98, 168.35.; 599 (M'"), 6 0 0 (M V i )
: Calculated for molecular fon-nula
C3oH23C1N604S.
;C , 60.15; H, 3.87;N, 14.03
: C 60.17; H 3.88; N 14.04%.
N ’-(2~Nitrobenzylidene)-2-[S'-((4~phenylphenoxy)mtethyl}~4~(4-chlorophenyl)-4H-
l,2,4-triazol-3-ylthio]acetohydrazide (120) Yield; 6 8 %; Wliite crystals, m.p,181-183
“C; Rf = 0.37 (CHCI3; MeOH; 9:1).
-1IR (KBr) cm'
'H NM R (300 MHz, CDCI3)
NM R (100 MHz, CDCI3)
ES-MS (w/j:)
Elemental Analysis
Calculated
Found
: 3058, 2932, 1682, 1607, 1522, 1492, 1378, 1352,
1235, 1093.
; 6 4.06 (s, 2H, S-CH2), 5.14 (s, 2H, O-CH2), 6.96
(d, J= 8.4 Hz, 2H), 7.32-7.56 (m, 12H), 8.01 (d,
/ - 6.8 Hz, IH), 8.14 (d, 6.9 Hz, IH), 8.19
(t,V = 7.5 Hz, IH), 8.31 (s, IH, N-H), 8.49 (s,
IH, Ar-H).
: 5 34.36, 59.87, 115.03, 124.63, 126.73, 126.90,
128.25, 128.29, 128.46, 128.71, 129.41, 129.82,
130.10, 130.37, 133.49, 134.96, 135.14, 136.95,
140.33, 140.66, 144.17, 148.05, 151.95, 152.30,
153.82, 156.70, 164.93, 169.35.
: 599 (M''), 601(M^+2).
: Calculated for molecular fonnula
C30H23CIN6O4S.
: C, 60.15; H, 3.87; N, 14.03
; C 60.19; H 3.85; N 14.04%.
100
Chapter II, Section 2
N'-(3,4,5-trimethoxybestzyUdene)~2-[5-{(4-phenylphenoxy)methyl}-4-(4~
chl0raphenyl)-4H'~l,2,4-triazol~3~ylthioJacetohydrazide (121) Yield, 75%; White
crystals, m .p.180-182 "C; Rf = 0.44 (w-hexane : etliylacetate; 4:6).
-IIR (KBr) cm
’H NM R (300 MHz, CDCI3)
‘^C NMR (100 MHz, CDCI3)
Maldi-MS (m/z) Elemental Analysis
CalculatedFound
; 3054, 2932, 1662, 1519, 1494, 1378, 1239, 1129, 1005, 836.
; 6 3.93 (s, 6H, O-CH3), 3.99 (s, 3H, 0-CH3),4.09 (s, 2H, S-CH2), 5.14 (s, 2 H,0 -CH2>, 6.95 (d, . / -7.2 Hz, 2H), 7.02 (s, 2H), 7.17 (s,lH), 7.35 (d, J = 7.2 Hz,lH), 7.42 (t, 7.5 Hz, 2H), 7.57-7.50(m, 8 H), 8.13 (s, IH, N-H).
: 5 38.58, 55.76, 59.21, 108.12, 114.48, 124.16, 125.93, 126.20, 127.43, 127.68, 127.90, 128.13, 129,39, 128.84, 134.23, 135.54, 139.61, 144.74,146.81, 152.23, 156.20, 163.06,167.94.
; 644 (M^), 645 (M'^+1).: Calculated for molecular formula C33H30N3CIN5O5S.
: C, 61.53; H, 4.69; N, 10.87: C 61.57; H 4.68; N 10.86%.
(E)-N'-(4-nitrobenzylidene)-2-f5-{(4-phenylphenoxy)methyl}~4-(4-methoxyphenyl)~
4H~l,2,4-triazol-3-ylthio]acetohydrazide (122) Yield: 80%; light yellow flakes, m.p,
168-170 "C; R f - 0.32 (CHCI3: MeOH; 9:1).
NO
N O ,
101
Chapter II, Section 2
IR (KJBr) cn f'
'H NMR (300 M Hz, CDCb)
13C NM R (100 MHz, CDCI3)
Maldi-MS (m/z)
Elemental Analysis
Calculated
Found
3058, 1684, 1603, 1241, 1107.
5 3.80 (s, 3H, O-CH3), 4.54 (s, 2H, S-CH2), 4.98
(s, 2H, O-CH2), 6.82 (d, J = 6 Hz, 2H), 7.20-7.0
5 (m, 3H), 7.40-7.29 (m, 13H), 8.00 (s, IH, N-
H), 9.88 (s, IH, OH).
5 34.36, 56.54, 59.86, 115.03, 124.62, 126.70,
126.90, 128.25, 128.27, 128.44, 128.70, 129.40,
129.82, 130.10, 130.36,133.46, 134.94, 135.14,
136.92, 140.30, 140.64, 144.17, 148.05, 151.94,
152.32, 153.82, 156.70, 156.84, 164.91, 169.32.
593 (M^), 594 (M'"+l).
Calculated for molecular formula C31H26N6O5S.
C, 62.61; H, 4.41; N, 14.13
C 62.65; H 4.42; N 14.10%.
(E)-N'-(4-ChlorohenzyUdene)-2-[5-{(4-phenylphenoxy)methyl}-4~(4- ethoxyphenyl)-
4H~l,2,4-triazol-3~ylthio]acetohydrazide (123) Yield: 78%; White crystals, m.p.166-
168 "C; M,-= O.SlCCHCls; MeOH; 9:1).
-IIR (KBr) cm'
’H NM R (300 MHz, CDCI3)
13C NM R (75 MHz, CDCI3)
HN,
NO
Cl
: 3056, 1679, 1603, 1454, 1252, 1 105.
: 6 3.91 (s, 3H, O-CH3), 4.62 (s, 2 H,S-CH2), 5.08
(s, 2H, O-CH2), 6.96 (d, J = 7.2 Hz, 2H), 6.99
(d, J - 7.2 Hz, 2H), 7.38-7.26 (m, 5H), 7.43 (t, J
= 6.9 Hz, 2H), 7.51-7.45 (m, 4H),7.68 (d, J =
7.5 Hz, 2H), 7.81 (s, IH, NH), 8.10 (s, IH,
CH).
; 5 36.56, 56.54, 59.86, 114.70, 115.12, 126.25,
126.47, 127.77, 128.09, 129.80, 127.93, 128.34,
128.46, 129.61, 130.79, 131.89, 132.13, 134.37,
102
Chapter II, Section 2
Maldi-MS {m/7)
Elemental Analysis
Calculated
Found
135,85, 139.91, 143.09, 146.76, 151.74, 151.39,
156.36,156.48, 164.05,168.50.
584 (M""), 585 (M^'+l).
Calculated for molecular formula
CsiHseClNsOaS.
C, 63.75; H, 4.49; N, 11.99
C 63.78; H 4.50; N 11.91%
2.4.2.2.2. Biological activity
2A.2.2.2.1. Anti-inflammatorv activity
All the targeted synthesized compounds were evaluated for their anti-inflammatory
activity against carrageenan-induced paw oedema in Albino Wistar rats weighing
150-200 g. Same method was used as discussed in section 1 of this chapter. The
results of anti-inflammatory activity are presented in table 2 . 2 and figure 2 .1 ,
2.4.2.2.2.2. Analsesic activity
The same method was used as given in section 1 o f this chapter. The results are
presented in table 2 3 and figure 2.2.
2.4.2.2.2.3. HistODatholosical Study
For the histopathological study, same methods was used as given in section 1 o f this
chapter, the results are presented in table 2.4 and figwre 2.3.
103
Chapter //, Section 2
2,4.23, ConclusionFrom the above results, it was concluded that two compounds 115 and 120 out o f
fourteen compounds from tliis series were found to possess better anti-inflammatory
activity (83.765 75,64 % and 82.84J 81.13 % inhibition) compared to the standard
drug Ibuprofen (78.89 and 80.10 Vo ) at 3 h and 5 h respectively. Compounds 115 and
120 showed comparable analgesic activity {(3.86, 3.88 and 2.85 sec.)} and {(3.80,
3.22 and 2.94 sec.)} with standard drug Ibuprofen (3.71, 3.95 and 2.76 sec.) at 30, 60
and 120 min. respectively. Compound 115 was found to be more potent causing no
ulceration than the standard drug Ibuprofen which showed ulceration.
104
tJJ?
f,l»{\m
m :
w C
Si * I ; t-.v, -).r'
i ? ?
% A.,.. ,::?i
:5 '/
1s-'
jA.:
. . i ii .
3 „ i „ t i i i t s i s t . '. ■ '■f V :■ r i; i PtS 1 “ 3
i-i < 1'. rt
ir
-i .ifi
ffj?
a
‘H NMR spectrum of compound 115
o
mo
m
i j p i i l p . |S-
tSfi .Hdt?.t (fj153'71
ii$i,'!>3.
i-t‘.» i-l
i-w :,s‘3 ' j.t->,o,|
!.»?<,as
..-■ i}a.3(y lilt. HI
i M . M ■ iM.if' t2»2H
i V | . V J ; y 2
IIM7HM.46
1 -o ■'« •■» I'3 r„ii C 3 * 1 5 ^ E» f cI ' ! t
■■ » U * itI I i l tS? wi a o X
; i*-j : <s ill s
g5; 1 S . ^ 1
... dldg ^S o S o .i i la ^ i s s g g s j s :
If I I i g ® i |
I ^W 'Qs k s i iS . |
1” !-...
s s gSSSH.».:"U5S!r , lS?iS?P.?r.SSJI5SS S;‘ tti ■;> C’*' ^:r‘ *-l’< c*.t> 's« »*•■ i * i
i s i l l i « r
( i~4r2') PI
t - i i j. r»-
,.'sBE
* C NMR spectrum of compound 115
gr
i
s *
J 8g; . K
fr:' ■'i'*
g
II»
■Ml ;I s S
J iS'.
i 's
r.
.„g
:. ® I: I '"
-:n’ s
>lf<'
I
I
i ;
H] | - g i 'fe
ss:S
A
el
ife2
Ig
,3;g
.mu. . -4
Mass spectrum of compound 115
T fa r» fiilte ii© e
i -
i -
c/>
'z/z
p
z x
o o<? o z
« ”0 » l» 'S f ci r
■S08‘'f*£-
s s > e t g -
___ „................. .. ^ . . - - - 3 ^
IR spectrum of compound 115
Chapter II
Section 3
Synthesis of 3-mercapto 1,2,4-triazole derived
mannich bases and their Biological Activities
Chapter II, Section 3
2,4.3»(l. Introduction
As discussed in introductory part of this chapter, mannich bases, biphenyl and
triazole derivatives exhibit different biological activities. In view of this mannich
bases of 1,2,4-triazoles and biphenyl systems, a focused library o f conjugates have
been synthesized and evaluated for their anti-inflammatory, analgesic, ulcerogenic
and antimicrobial activities. The synthetic route is given below.
SCHEME III
N“ N
N SH
63 R1 = 4 -0 IV le
65 R1 = 4-CI
68 R1 = 2-M e
Ri
different 2“ amines, HCHO
Alcohol, stirring, 24 h
Q
Rg = H3C-N,/ \\__/N-5
R2
124-128
Q f
The above synthesized 3-mercapto-l,2,4-triazoles (63, 65, 68) were treated with
formaldehyde and secondary amines under mannich reaction conditions to form the
novel mannich bases (124-128). Various secondary amines including morpholine,
pyrrolidine, N-methylpiperazine were used to check the generality of the reaction and
also to bring the functional group variations. All the synthesized compounds have
been completely characterized on the basis o f their detailed spectral data and the
entire novel compounds of this series were evaluated for their biological activities.
The physical data are given in table 3.1.
105
Chapter II, Section 3
Table 3.1 Physical data of 3-mercapto 1,2,4-triazole derived mannich bases
Compoundno.
Structure % Yield m.p (°C)
124 .n-n' - OO - O o - ^ ^ - ^ s
9G!
58 180-182
125
p°'CHo
54 184-186
126 80 174-176
127
o
64 132-134
128 52 126-128
2.43 J . Results and Discussion
2 .4 .3 .U . Analytical
Five novel compounds 124-128 were synthesized starting from triazole 63, 65 and 68
as outlined in scheme-III. Reaction of triazole 63, 65 and 68 with formaldehyde and
secondary amines in absolute alcohol afforded mannich bases (124-128). The 1,2,4-
triazole ring exists in two tautomeiic forms, under the mannich reaction conditions,
the -NH proton of the triazole ring will participate in the reaction and will fonn the
final mannich bases (124-128). Fonnation of mannich bases was confirmed from 'H
106
Chapter II, Section 3
NMR spectrum exhibiting a singlet at 5 4.47”4.93 ppm due to methylene proton (N-
CH2-N) integrating for two protons. The '^C NMR spectrum o f compounds 124-128
displayed peaks in the range 3 68.76-68.82 for the carbon bonded with two nitrogen
(N-CH2-N) and the peaks in the range of S 170-172 ppm for the carbon attached with
sulphur (C=S) in the triazole ring and the spectral data are in line with Barbuceanu et
al. [9] which confirms the formation of the compounds 124-128.
All the newly synthesized compounds 124-128 were screened for their biological
activities (anti-inflammatory, analgesic, ulcerogenic and antimicrobial).
2.4.3.1.2. Biological Activity
2,4.3.1.2.1. Anti-inflammatory activity
The results of anti-inflammatory activity are summarized in table 3.2 and figure 3.1.
Among five compounds from this series, two compounds 125 and 128 were found to
be active and exhibited significant activity (54.54, 79.69 % and 60.06, 84.08 %
inhibition) compared to the standard drug Ibuprofen (78.93 and 82.58 %) at 3 h and
5 h respectively. Compound 128 was found to be more potent than the standard dmg
Ibuprofen. All data were analyzed by one-way ANOVA test followed by Dunnett’s
test in carrageenan induced rat paw oedema.
Table 3.2. Anti-inflammatory activity of 3-mercapto 1,2,4-triazole derived mannich bases
Com pds. Change in paw oedema vo lum e (m L )
A fte r d ru g trea tm en t
A n ti- in f la m m a to ry a c tiv ity
% In h ib it io n
3ii Sh 3h 5h
124 0.387±0,043” 0,367±0.047” 41.36 44.89
125 o.sooiG.oee*** 0.266±0.088**‘ 54.54 60.06
126 0 ,410±0,072"’ 0,436±0.074 37.87 34.53
127 0.566±0,042'"‘ 0.516±0.119"'^ 14.24 22.52
128 0.134±0.055**‘ 0.106±0.047*” 79.69 84.08
Ibuprofen 0 ,f3 9 ± 0 ,0 2 r ” 0 , l l6 ± 0 .0 5 r " 78.93 82.58
Control 0.660±0.042 0.666±0.088 — —
Data analyzed by one way A N O V A fo llow ed by D unnett’ s ' t ' test (n=6), *p<0.05, *p<0.01 & **‘p<0.001 s ign ifican tly d ifferent from standard; ns, not significant,
107
Chapter II, Section 3
90
80
70
_ 60 O£ SO
J 40
^ BO
20
10
0
Anti-inflam m atory activity %
M B li U 5 h
ir124
\ i4125 126 127
Compounds
128 Ibuproftn
Figure 3.1. Anti-inflammatory activity of 3-mercapto 1,2,4-triazole derived mannich bases
2.4.3.I.2.2. A n a ls e s ic A c t iv ity
The results o f analgesic activity are summarized in table 3.3 and figure 3.2. Both the
tested compounds, 125 and 128 showed comparable analgesic activity (reaction time)
with the standard drug (Ibuprofen). All data were analyzed by one-way ANOVA test
followed by Dunnett’s test.
Table 3.3 Analgesic activity o f 3-mercapto 1,2,4-triazole derived mannich bases
Compounds Mean value of TaU Flick Latency (sec)±SEM
0 min. 30 min. 60 min. 120 min.
125 2.08±0.32 3.80±0.233* 3.22±0.264* 2.94±0.232“
128 1.93±0.303 3.86±0.346* 3.88±0.243* 2.85±0.237“
Ibuprofen 1.98±0.307 3.71±0.38S’ 3.95±0.473* 2.76±0.212“*
Control 2.03±0.330 1.94±0.236 2.02±0.192 2.85±0.237
*p<0.001 significantly different from standard; ns, not significant.
108
Chapter II, Section 3
Figure 3.2. Analgesic activity of 3-mercapto 1,2,4-triazole derived mannich bases
2.4.3.I.2.3. H is to p a th o lo s ic a lS tu d y
Histopathological results revealed that compounds 125 and 128 did not cause any
gastric ulceration and disruption of gastric epithelial cells at the given oral doses when
compared to standard Ibuprofen. Hence gastric tolerance o f these compounds was
better than that o f Ibuprofen indicating that carboxylic group present in Ibuprofen is
responsible for ulceration. Stomach wall of Ibuprofen treated group at low power
(lOx) photomicrograph showed damage o f the mucosa and the sub mucosa. Stomach
wall o f the same section at high power (40x) photomicrograph showed desquamated
epithelial cells in the lumen. The results are shown in table 3.4 and figure 3.3.
Table 3.4. Haematoxylin and Eosin immunohistochemical staining o f gastric ulcers
after ulcer induction in rats.
Compds. Surface Epithelial
Damage
Submucosal
Damage
Deep Mucosal
Damage
Muscular
Layer
Damage
125 - - - -
128 - - - -
Control - - - -
Ibuprofen -H -+ -H -l- - -
Indomethacin +++ +++ +++ +
indicates no ulceration; +, ++, +++ indicates increased ulceration.
109
Chapter II, Section 3
125 iWK^LUMCN
4'L-i
125 (lOX)C WMC M
Ibiipi ofni (lOX)LU M C N
i-". #- -:
(oiitiol (lOX)
i:5 (40X)
*■ I . f ,» ' I f /
128 (40X)
/l u m b n
Ibnpi ofni (40X)L U M B M
_ ' ^ V i 7tt<
ronti ol (40X)
Figure 3.3 Haematoxylin and Eosin immunohistochemical staining of gastric ulcers after ulcer induction in rats.
110
Chapter II, Section 3
2 .4 3 .1 ,2 .4 . Antimicrobial Activity
All the newly synthesized compounds from this series 12 4 -12 8 were tested against
various microbial strains at the concentration of 200 fa,g/disc, 100 )o,g/disc for their
antimicrobial activity. The results are summarized in ta b le 3 .5 . Some o f the
compounds from this series showed moderate to good activity against bacterial stains.
Compounds 124, 125 and 128 showed good activity against S. aureus.
Table 3.5. Antimicrobial activity o f 3-mercapto 1,2,4-triazole derived mannich bases
Compds Antifiingal Activity (zone of inhibition in mm) Antibacterial Activity (zone of inhibition in mm)A.Jlavus 200 (100) (!g/disc
A.niger 200 (100) (.ig/di.sc
C .a lh i c a n s
200(100)fig/disc
C.krusei200(100)jig/disc
K.pneiimonia if 200 (100)
Hg/disc
P .a c m g i n o s a
200(100)jiig/disc
E .c o l i
200(100)Hg/disc
S.aureus200(100)
fig/disc
124 8± 0.6
(6±0.4)
8±0.6
(-)
6±0.4
(-)
9±0.6
(6±0.4)
8±0.4
(-) ■
12± 1.2
(6±0.3)
125 6± 0,6
(-)
8±.7
(-)
10± 0.8
(8± 0 .6)
9±0.4
(-)
“
■
12± 1.2
(6± 0 .6 )
126 6±0.4
(-)
8± 0.8
(-)
10± 1,2
(8±0.5)
8± 1.2
(8± 0 .6)
10±0.9
(6±0,4)
8± 0.6
(-)
10± 0.8
(8±0,7)
127 12± i .6
(6±0.4)
10± 1.6
(-)
10± 1.6
(6±0 ,8)
10± 1.6
(8± 1.2 )
10± 1,6
(5±1.6)
8± 1.6
(4)
8± 1.6
(6± 1.6)
10± 1.6
(6± 1,6)
128 10±!,2
C8±0.6)
11±1
(6±0.3)
8± 0.6
(-)
8±0.6
(6± 0 .6)
10± 1.2
(6±0.4)
S±OA
(-)
lO il .2
(6± 0 .6)
12±0.9
(6±0,S)
Flucon. 22± 1.8
(12±0,9)
1 6 il
( 10±0 .6 )
M i l . 2
(lO iO .l)
lOi-1.2
(8±0,7)
N T TMT N T N T
Control - - - - - - - -
Am oxcil N T N T N T NT 16±1
( 12± 0 .8)
14±0.8
(I0±0,5)
12±0.7
(8±0,4)
I 2±0.6
(6±0.3)
Flucon: Fluconazole; A m oxcil.: Am oxcillin ; N T : not tested; no zone o f inh ib ition.
I l l
Chapter II, Section 3
2.43.2„ Experimental
2.4.3.2.1. Chemistry
General procedure fo r synthesis o f 4-(substitiitedphenyl)-3-(amme-4-ylmethylthw)~
S~(4-hiphenyloxymethyl)-4H-I,2,4-triazole (124-128)
To 50 mL absolute alcohol was added appropriate triazole 63, 65 and 68 (5 mmol)
formaldehyde (5 mmol) and appropriate amine (5 mmol), the reaction mixture was
stirred for 24 h, and allowed to stand over night in refrigerator. The precipitate so
obtained was filtered, washed with cold alcohol and recrystallized from alcohol.
5-l(4-Eiphenyloxy)methyl]-4-(4-chlorophenyl)-2-(morpholinomethyl)-2H-l,2,4~
triazole~3(4H)-thione (124) Yield: 58%; White crystals, ni.p. 180-182 "C; Rf == 0,62
(CHCI3: MeOH; 9:1).
-1IR (KBr) cm’
‘H NMR (3§0,MHz, CDCI3)
13C NMR (100 MHz, CDCI3)
ES-MS {m/z)
Elemental Analysis
Calculated
Found
QV N/ / -N
2909, 1608, 1381, 1115.
5 2.86 [t, J= 4.5 Hz, 4H, N-(CH2)2] 3.72 [(t, J =
3.9 Hz, 4H, 0 -(CH2>2 ], 4.93 (s, 2H, N-CH2-N),
5.18 (s, 2H, O-CH2), 6.89 (d, = 8.7 Hz, 2H),
7.29-7.86 (m, 11H).
5 50.74, 59.99, 66.81, 69.86, 115.11, 126.76,
127.03, 128.34, 129.26, 129.95, 132.08, 135.51,
136.29, 140.22, 146.12, 156.41, 170.72.
493 (M' ).
Calculated for molecular fonnula C26H25N4O2S.
C, 63.34; H, 5.11; N, 11.36.
C, 63.52; H, 5.18; N, 11.41%.
112
Chapter II, Section 3
5-f(4~Biphenyloxy)methylJ-(4-methoxyphenyl)-3~(N~methyl~piperazme~4-
ylmethylthio)-4H-I,2,4-triazole (125) Yield: 54%; White crystals, ni.p. 184-186 ”C;
R f - 0.59 (CHCI3: MeOH; 9:1).
- 1IR (KBr) cm'
‘H NMR (200 MHz, CDCI3)
NMR (100 MHz, CDCI3)
Maldi-MS {m/z) Elemental Analysis
Calculated
Found
C H ,
CHo
: 2907, 1557, 1326, 1109.
: 5 2.25 (s, 3H, N-CH3), 2.5l(t, J = 9 Hz, 4H),
2.71 (t, J = 7.8 Hz, 4H), 3.81 (s, 3H, O-CH3),
4.49 (s, 2H, N-CH2-N), 5.07 (s, 2H 0-CH2),6.96
-7.11 (m, 4H), 7.36-7.60 (m, 9H).
: 5 44.14, 49.26, 56.24, 57.15, 68.45, 68.82, 15.45,
126.92, 127.62, 127.89, 128.69, 129.61, 129.64,
130.29, 135.69, 149.66, 156.68, 155,64, 163.49,
171.82.
501 (M^), 502 (M'^+1).
Calculated for molecular fomiula C28H31N5O2S.
C, 67.04; H, 6.23;N, 13.96.
C, 67.01; H, 6.21; N, 13.93%
5-[(4-Biphenyloxy)methyl]-4-(4-methoxyphenyl)-2-[(pyrrolidin-l~yl)methyl]-2H~
l,2,4-triazole-3(4H)-thione (126) Yield: 80 %; White crystals, m,p. 174-176°C; Rf
0.50 (CHCI3: MeOH; 9:1).
IR (KBr) cm - 1
CH,
: 2907, 1607, 1516, 1316, 1104.
113
Chapter II, Section 3
'H NMR (200 MHz, CDCI3) ; 5 1.53 (t, J - 7.5 Hz, 4H, CH2-CH2), 2.24 ( t , ./ -
8.0 Hz, 4H, N-CH2), 3.82 (s, 3H, O-CH3), 4.47
(s, 2H, N-CH2-N), 5.22 (s, 2H, O-CH2), 7.54 -
6.91 (m, 13H, Ar-H).
‘■'’C NMR (100 M H2, DMSO-dfi) ; 8 26.41, 52.42, 56.24, 68.45, 68.76,15.42,126.75,
126.89, 127.79, 128.69, 128.89, 129,58, 129.67,
130.27, 133.89, 135.65, 149.68, 155.76, 163.54,
172.86,
Maldi-MS (m/z)
Elemental Analysis
Calculated
Found
472 (M^), 473 (M'^+1).
Calculated for molecular formula C27H28N4O2S.
C, 68.62; H, 5.97; N, 11.85.
C, 6 8 .68; H, 5.99; N, 11.87%.
5-f(4-Biphenyloxy)methyl]-4-(phenyl)-2~(morpholinomethyl)-2H-l,2,4~triazole~
3(4H)~thione (127) Yield; 64 %; White crystals, m.p. 132-134 °C; Rf == 0.53 (CHCI3:
MeOH; 9:1).
N- N
IR (KBr) cm '
NMR (200 MHz, DMSO-dg)
v i:2912, 1607, 1324, 1103.
: 5 2.90 (t, J = 7.0 Hz, 4H), 4.76 (t, J = 6.7 Hz,
4H), 4.97 (s, 2H, N-CH2-N), 5.22 (s, 2 H,0 -CH2)
6.93 (d ,/ = 8.8 Hz, 2H), 7.40-7.54 (m,12H).
NM R (100 MHz, DMSO-df,) : 5 49,23, 64.86, 68.45, 68.76,64, 115.42, 126.89,
127.79, 128.69, 128.89, 129.58, 129.67, 130.27,
133.89, 135.65, 149.68, 155.76, 163.49, 170.56. 458 (M""), 459(M^+1).
Calculated for molecular formula C26H26N4O2S.
C, 68,10; H, 5.71;N, 12.22
C, 68.07; H, 5.69; N, 12.20%
Maldi-MS {m/7)
Elemental Analysis
Calculated
Found
114
Chapter II, Section 3
5~f(4-Biphenyloxy)methyl]~4~(2->methylphenyl)~2-(morpholmomethyl)-2H~l,2y4-
triazole-3(4H)-thione (128) Yield: 52%; White crystals, m .pJ26-128 "C; Rf = 0.51
(CHCI3: MeOH; 9:1).
V jCH.
-1IR (KBr) cm
’H NMR (300 MHz, CDCI3)
NMR (100 MHz, DMSO-ds)
MaMi-MS {m/z)
Elemental Analysis
Calculated
Found
2.4.3.2.2. Biological activity
: 2917, 1516, 1324, 1108.
: 5 2.50 (s, 3H, Ar-CHs), 2.72 (t, J - 6.9 Hz, 4H),
3.57 (t, / = 6.7 Hz, 4H), 4.93 (s, 2H, N-CH2-N),
5.23 (s, 2H, O-CH2), 6.90 (d, J = 8.7 Hz, 2H),
7.30-7.58 (m, IIH).
: 5 14.42, 49.24, 64.87, 68.56, 68.79, 115.43,
26.88, 126.97, 126,98, 128.81, 128.86, 129.57,
129.63, 130.26, 134.43, 135.66, 136.84, 149.66,
155.86, 163.51, 170.58.
472 (M^), 473(MV1)
Calculated for molecular formula C27H28N4O2S.
C, 68.62; H, 5.97; N, 11.85
C, 68.65; H, 5.95; N, 11.83%.
2.4.3.2.2.1. Anti-inflammatory activityAll the synthesized compounds were evaluated for their anti-inflammatory activity
against carrageenan-induced rat paw oedema in albino Wistar rats weighing 150-200
g. Same method was used as discussed in section 1 of this chapter. The results of anti
inflammatory activity are presented in table 3.2 and figure 3.1.
2.4.3.2.2.2. Analsesic activity
The analgesic activity was detennined using tail flick method in Albino Wistar rats
weighing 150-200 g. The same method was used as given in section-1 o f this Chapter.
The results are presented in table 3.3 and figure 3,2.
115
Chapter II, Section 3
2.4.3.2.2.3, HistoDatholo2icai study
For the histopatliological study, same method was used as given in section 1 o f this
chapter. The results are presented in table 3,4 and figure 3.3,
2.4.3.2.2.4, Antimicrobial activity
All the synthesized compounds were evaluated for their antimicrobial activity against
the various bacterial and fungal strains. Same methods were used as given in section-1
of this chapter. The results are presented in table 3.5.
116
Chapter II, Section 3
2,43.3, ConclusionThe designed molecules were successfully synthesized using mannich reaction. All
the novel mannich bases synthesized were tested for their anti-inflammatory,
analgesic, ulcerogenic and antimicrobial activities- From the results, it was concluded
that two compounds 125 and 128 showed good anti-inflammatory activity (54.54,
79.69 at 3h and 60.00, 84.08 % at 5 h respectively) compared to the standard drug
ibuprofen. Among the tested compounds, compound 128 showed moderate analgesic
activity without any gastric ulceration whereas the standard drug ibuprofen caused
ulceration. From antimicrobial studies it was concluded that the compounds from this
series showed moderate activity against the tested bacterial strains.
117
«
i l to I^ ‘
m
I S ,
i W
««
1.01 ■w
1
■<.-”• 3 J J 6
S..723 J.7«i s n'%M2 2,PS
>ife 4a= ?•' 4 > -ta f ^ >-t ^ ‘h
i / t r t i '■ i e f * > ‘ K.l »>
13*'* »‘ l ?. n j. 'ifi
s
1«?
1If-
O
P ^
':■ ' ■ .■■ ^ A - ■ ■ '■
r p ' i mwr- f »■•• #-•»tr. ».V i.*-.« » '»
k-.H ,
i f 1 ■ t» ■!
" f, i
m i'.".V :
'£h '. .r .r - .“V-’r
t»/ w t j -JZ’I t % % "■
U ! <t- t fA ‘-Z »-\ U wti ii't<’< 7>
m ■ tti: ■P><{-.M.'Jki C4 i • •‘-1-;3
*ily ^
i "
■IS
'H NMR spectrum of compound 124
MOO
©3O
<3?O
o
MO
oo
0 3o
o
I7U.72
128.34127.03126.76IIJ.ll
. 77,43 77.00 7S.38 -— 6')M ■•—'-<>6,8!
----- -
-------- 30.7-1
10 8:1 l ig a ;
■.' O ' . j ' .
> <j> Q O ih-*; )-* nip 0V«? 0 KC3iCn::ii'
03 Q Ow o ,.lio <>.o r\i> I'j Kj cao Q -J U» i<'*' »vjVP o'«£>'^ <?i' .<N> >a>- -tft'i'''. »£» 0 ' 0 'c > 'f ' 0 3 i J i t P iJS^'-OV
,.VC3' C ? C> V . O O Q........)'.o o "" ................
g., iv.'rrr:H ■
w -w'ii ci. I' vca. .rc'js;.'■f& (T> «a crt ,(1j M :W:'O. a.-'D;; ;<». :■■ ■. a ■'■;■■■.■■
'■•Mi;
‘ ■C NMR spectrum of compound 124
S3JL ^
8
§
J , . , ....
2o
sfels
iU5
Ia
& : mk
z ^ : s
CO
£!
Ilf
? ri
Mass spectrum of compound 124
W ^ m
/CO
o
IR spectrum of compound 124
Chapter II
Section 4
Synthesis of Quinoline based 1,2,4-triazole
derivatives and their Biological Activities
Chapter II, Section 4
2.4,4.0. Introduction
In view of the biological importance 1,2,4-triazoles and quinoline as discussed in
introductory part of this chapter, we intended to conjugate two moieties in order to
develop novel antiinflammatory and antimicrobial agents. A focused library of
conjugates have been synthesized and evaluated for their anti-inflammatory,
analgesic, ulcerogenic and antimicrobial activities. The synthetic route is given below.
SCHEME-IV
129
ClCHzCOOEt di-y acetone K2CO3, reflux
CH2COOC2H5
Abs. alcohol NH2NH2.H2O
130
CH2CONHNH2O
131
Abs.alcoholArylisothiocyanatereflux
O'CH2CONHNH
'= S NH
132-135
Abs.alcohol TEA, reflux
N -n
fsj' SH
Ri
Dry acetoneK2CO3Phenacylbromidereflux
136-139 140-149
Reaction of 8-hydroxyquinoline (129) with ethyl chloroacetate in presence of
potassium carbonate in anhydrous acetone afforded Ethyl 2-(quinolin-8-yloxy)acetate
(130) which readily yielded 2-(quinolin-8-yloxy)acetohydrazide (131) by refluxing
with hydrazine monohydrate in absolute alcohol. Compound 131 was converted into
corresponding thiosemicarbazide (132-135) by reacting with different substituted aryl
isothiocyanate in absolute alcohol. The thiosemicarbazides (132-135) were cyclised
118
Chapter II, Section 4
into respective 4-substituted phenyl-5-[(quinolin-8-yloxy)methyl]-4H-l,2,4-triazoie-
3-thiol (136-139) using triethyl amine in absolute alcohol. Finally S-alkylation of 3-
inercapto-l,2,4-triazoles with substituted phenacyl bromides leads to the formation of
target molecules 140-149. All the synthesized compounds have been completely
characterized on the basis of their detailed spectral data and the entire novel
compounds of this series were evaluated for their biological activities.
The physical data o f all the final compounds are given in table 4.1.
Table 4.1 Physical data of quinoline based 1,2,4-triazole derivatives.
Compounds Structure % Y e ild m.p, ( "O
140
'C l
54 170-172
141N - X ,
0 OH
62 228-230
142 72 218-220
143 63 188-190
144
Cl
72 180-182
145
Cl
68 168-170
146
° 'C H j
49 172-174
147 62 160-162
119
Chapter II, Section 4
148i;J 's-vAJ 68 230-232
149
c t S 6 °74 158-160
2.4«4.1. Results and discussion
2.4.4.1.1. Analytical
A focused library of new compounds 140-149 was synthesized starting from 8-
hydroxy quinoline as outlined in schenie-IV. O-alkylation of 8-hydroxy quinoline
with a-chloro ethylacetate was confinned by the 'H NMR spectiTtm. The appearance
of two signals at 5 1.52 (t, J = 5.6 Hz, 3H) and 3.88 (q, J ~ 5.8 Hz, 2H) supported the
presence of ethyl gi'oup and appearance of strong absorption at 1678 cm”' in IR
spectrum confirms the formation of ethyl 2-(quinolin-8-yloxy)acetate (1 3 0 ).
Compound 130 readily yielded 2-(quinolin-8"yloxy)acetohydrazide (1 3 1 ) by its
reaction with hydrazine monoliydrate. The formation of this hydrazide was confirmed
by the presence of signals due to NH-NHj group at 5 4.09 (br, s, NH2, 2H) and 9,71
(s, N-H, IH) and absence of signals due to ethyl group. Compound 1 3 1 was then
converted into corresponding thiosemicarbazides (1 3 2 -1 3 5 ) which was confirmed by
the presence of extra signals in aromatic region due to phenyl ring o f isothiocyanate
along with signals at 5 8.44-8.62 (CSNH), 9.08-9.21 (CONH) and 8.02-8.08 ppm (Ar-
NH-) in the 'H NMR spectrum. It is interesting to note that thiocarbonyl compounds
are present in their thionethiol tautomeric forms in solution as indicated by their IR
and 'H NMR spectra [56]. These tautomeric fonns are also present in dimethyl
sulfoxide as suggested by NMR spectral data. It is already reported that the acid (HCl,
H2SO4) mediated cyclization of thiosemicarbazides leads to the fomiation of 1,3,4-
thiadiazoles [57]. Cyclization of compounds 1 3 2 -1 3 5 to 1 3 6 -1 3 9 was confirmed from
the IR and NMR spectral data. Absorption bands in the region 1601-1605 cm'' (C=N
of triazole ring stretching) and the presence of signal as singlet corresponding to thiol
120
Chapter II, Section 4
proton (-SH) in the range of 8 14.10-14.18 ppm in ‘H NMR spectrum confirmed the
formation of 3-mercapto-l,2,4-triazole.
Formation of alkylated derivatives 140-149 from the cyclized compound 136-139
were confirmed from the disappearance of -SH signal. Instead, new signals that
originated from methylene (4.88-5.02, -S-CH2) and carbonyl group (190-193, C=0)
were observed in the 'H and '^C NMR spectra of compounds 140-149. The -SH
proton is acidic enough and some substitution reaction could be achieved on this
group in the presence of a base [16, 34].
All the newly synthesized compounds 140-149 were screened for their biological
activities (anti-inflammatory, analgesic, ulcerogenic and antimicrobial activities).
These novel triazole derivatives have shown varied therapeutic efficacy in vitro and
in vivo.
2,4.4.!.2. Btologlcal Activity
2.4.4.1.2.1. Anti inflammatory activity
Compounds 142, 143 and 149 from S-substituted phenacyl 1,2,4-triazole exhibited
significant anti-inflammatory activity with 6 8 .4 O5 63.34, 68.40 % and 72.33, 71.87,
75.00 % inhibition at the end of 3 h and 5 h respectively compared to standard drug
hidomethacin (55.29 and 69.50 %). The active compounds have been further
investigated for their analgesic activity and ulcerogenic studies. Among the above
synthesized library of compounds, compound 149 (68.40 and 75.00 % inhibition at
3 h and 5 h respectively) showed highest inhibition in rat paw oedema. The results are
given in table 4.2 and figure 4.1.
121
Chapter II, Section 4
Table 4.2. Anti-inflammatory activity of quinoline based 1,2,4-triazole derivatives.
Compounds Change in paw oedema volume (mL) After drug treatment
Anti-inflammatory activity % Inhibition
3h 5h 3h 5h
140 0.533± 0.03“ 0.433 ±0.07"* 15.79 27.83
141 0.316±0.04“ 0.283±0.06*’ 50.07 52.83
142 0.200 ±0.16“ * 0.166±0.04*** 68.40 72.33
143 0.232 ± 0.18*** 0.178 ±0.08*** 63.34 71.87
144 0.416±0.07“ 0.400±0.07"^ 34.28 33.33
145 0.283±0.04** 0.200±0.03** 55.29 66.66
146 0.616±0.08"* 0.466±0.12“ 20.00 22.33
147 0.633±0.08"* 0.483±0.08“ 00.00 19.50
148 0.533±0.10”® 0.466±0.09“ 15.79 22.33
149 0.200±0.05*** 0.150±0.03*** 68.40 75.00
Control 0.633±0.05 0.600±0.03 — —
Indomethacin 0.283±0.03” 0.183±0.30*’* 55.29 69.50
*p<0.001 significantly different from standard; ns, not significant.
ISR
Anti-inflam m atory activity
■ 3 h r U 5 h r
Com pounds
Figure 4.1. Anti-inflammatory activity of quinoline based 1,2,4-trijizole derivatives.
122
Chapter II, Section 4
2.4.4.I.2.2. A n a ls e s ic a c t iv ity
The results o f this series o f compounds, v iz . S-substituted phenacyl 1,2,4-triazoles
(140-149) conclude that none o f the compounds is superior than the standard drug
Indomethacin but all the compounds showed moderate to comparable activity.
Among the tested compounds, compound 143 and 149 showed comparable analgesic
activity (4.58, 5.27 and 4.88, 5.41) with the standard drug Indomethacin (5.79, 6.42)
at 30 min. and 60 min respectively. The results are summarized in table 4.3 and
figure 4.2. All the data were analyzed by one way ANOVA test followed by
Dunnett’s test.
Table 4.3. Analgesic activity o f quinoline based 1,2,4-triazole derivatives.
Compounds Mean value of Tail Flick Latency (sec)±SEM
0 min. 30 min. 60 min. 120 min.
142 3.50±0.247 4.38±0.287* 4.77±0.264* 3.50±0.247"‘'
143 3.46±0.247 4.58±0.399* 4.88±0.285‘ 3.46±0.247"*
149 3.60±0.450 5.27±0.365*‘ 5.41±0.570** 3.60±0.450“
Control 2.89±0.170 2.93±0.283 3.16±0.254 2.89±0.170
Indomethacin 3.38±0.321 5.79±0.360’“ 6.47+0.527’*’ 3.38*0.321"“
*p<0.001 significantly different from standard; ns, not significant.
s
7
6
A S
E
I 3
ar2
1
0
142
A n a lg e s ic activ ity
•^Omin ■30m in 'JSOmin ' ’ 120min
T T
1
L.\
143 149 control lndom«thacin
Compounds
Figure 4.2. Analgesic activity o f quinoline based 1,2,4-triazole derivatives
123
Chapter 11, Section 4
2.4.4=1.2.3. Ulcerocenic study
Screening of stomach wall of Indomethacin treated group at low power (lOx)
photomicrograph showed ulceration and necrosis of the mucosa, the necrosed area is
marked by arrows. Stomach wall of indomethacin treated high power (40x)
photomicrograph of the same section showed necrotic epithelial cells surrounded by
fibrinoid matrix of the mucosa, which indicated significant surface epithelial damage,
submucosal damage, deep mucosal damage and slightly mucular layer damage.
Whereas the test compound 142 treated animal showed significant surface epithelial
damage, slightly submucosal damage and no damage of deep mucosal and muscular
layer. Compound 149 treated animal exhibited no damage of any layer. All the results
are presented in table 4.4 and Figure 4.3.
Table 4,4. Histopathological studies of quinoline based 1,2,4-triazole derivatives
G roup Siii-face
E p ithe lia l
Damage
Submucosal
Damage
Deep M ucosal
Daniage
M uscu la r
L aye r
Damage
142 +++ ++ ~ “
149 -
■ ■ ■
Control “
■ ■ ■
Ibuprofen +++ +++ “ "
Indomethacin +++ +++ +++ +
No damtige; +, ++, +++; indicates increasing degree o f damage
Continued.
124
Chapter II, Section 4
IndomMfaacm, 10 X Jndom vH tariii, -40X
Figure 4.3 Haematoxylin and Eosin immunohistochemical staining o f gastric ulcers after
ulcer induction in rats.
125
Chapter II, Section 4
2.4,4.1.2.4. Antimicrobial activity
The antifungal results showed that some of the compounds exhibited good antifungal
activity against A. niger and A. Jlavus. Compounds 140 and 141 showed good growth
of inhibition against A. niger. Compounds 143, 144 and 149 were active against A.
flavus. It was observed that the maximum antimicrobial activity was observed by the
tested compounds having Ri= 3-Cl, 4-Cl, methoxy group in phenyl ring of the 1,2,4-
triazole system. Replacement of chloro group in the phenyl ring diminishes the
antifungal activity
The antibacterial results showed that some of the compounds displayed good
antibacterial activity against K. pneumoniae and P. aeruginosa. Compounds 142 and
144 showed good growth of inhibiton against both the bacterial strains. It can be
concluded that compound 143 was superior to positive controls against P. aeruginosa.
It was observed that the maximum antibactei'ial activity was displayed by the tested
compounds having 4-Cl, 4-Br groups present in phenyl ring of the 1,2,4-triazole
system. Replacement of chloro group in the phenyl ring decreases the antibacterial
activity.
Table 4.5. Antimicrobial activity of quinoline based 1,2,4-triazole derivatives.
CompdsA n tifu n g a l A c t iv ity
(zone of inhibition in mm)A n tib a c te ria l A c t iv ity
(zone of inhibition in mm)
A.flavus 200(100) i^g/disc
A. niger 200 (100) |4,g/disc
Kpneum oniae 200 (100) jag/disc
P.aeroginosa 200 (100) |ag/disc
140 10 (8) 14 (12) 14(8 ) (-)
141 12(8) 18 (11) 15 (10) (-)
142 10 (8) 8 (6) 12 (7) 14 (8)
143 14 (10) 12(10) 9 (5 ) 20 (12)
144 14 (12) 11(8) 18 (10) 14 (8)
145 (-) 6 10 (6) 14 (8 )
146 (-) (-) 6 (-) 8 (6)
147 12(10) 9 (6) 10(10) 12(8 )
148 10(9) 9 10(8) 6149 14 (12) 10 (8) 14 (10) 6
DMSO - - -
Fluconazole 14 12 NT NT
A m o x ic iliin NT NT 12 10
- : no zone o f inh ib ition ; nt: not tested
126
Chapter II, Section 4
2A A 2 , Experlifieiital2.4.4.2.L Chemistry
All chemicals (reagent grade) used were commercially available. Melting points were
measured on a VEEGO-VMP-DS melting point apparatus and are uncorrected. 'H
NMR was recorded on a Biuker DPX 500, 400, 300 instruments in CDCI3/DMSO-df,
using TMS as internal standard for protons. 'H NMR chemical shifts and coupling
constants J are given in ppm and Hz respectively. Mass spectra were recorded on a
Jeol JMS-D 300 instrument fitted with a JMS 2000 data system at 70 eV and Maldi-
MS, Mass-spectrometric (MS) data is reported in m/z. Elemental analysis was carried
out using Elemental Vario EL III elemental analyser. Elemental analysis data is
reported in % standard.
General procedure fo r synthesis o f Ethyl 2-(quinolin-8-yloxy)acetate (130) Yield:
75%; Light yellow crystals, ra,p= 51-52 "C, Rf = 0.42 («-hexane : ethylacetate; 6:4).
- 1IR (KBr) cm
’H NMR (300 MHz, CDCI3)
FAB-MS (m/z) Elemental Analysis
Calculated
Found
3052, 2921, 1678, 1109.
8 1,52 (t, 3H, / = 5.6 Hz), 3.92 (q, 2H, J = 5.8
Hz), 5.29 (s, 2H, O-CH2), 7.02 (d, 2H, ./ = 8 . 6
Hz), 7.24(s, IH, 7.8 Hz), 7.36-7.54 (m, 3 H,
Ar-H).
231 (M' ), 232 (M'^+1).
Calculated for molecular formula C 13H13NO3 .
C, 67.52; H, 5.67; N, 6.06.
C, 67.53; H, 5.68; N, 6.07%.
2-(quinolin-8-yloxy)acetohydrazide (131) Yield: 84%; Light yellow crystals, m.p,
150-152 *’C, Rf = 0.25(CHCl3: MeOH; 9:1).
.N X O
IR (KBr) cm'-1 3352,3054, 2921, 1684, 1109.
127
Chapter II, Section 4
’H NMR (300 MHz, CDCI3)
FAB-MS {m/z)
Elemental Analysis
Calculated
Found
5 4.09 (s, 2H, NH2,), 4.87 (s, 2H, O-CH2), 9.71
(s, IH, N- H), 8.94 (d, 7 = 3 .9 Hz, IH), 8.20 (d,
8,4 Hz, IH), 7.17-7.60 (m, 4H, Ar-H).
217(M"),218(M'"+1).
Calculated for molecular formula C 12H12N2O2.
C, 66,65; H, 5.59; N, 12.96
C, 66.71; H, 5,56; N, 12.92%.
General procedure fo r synthesis o f thiosemicarbazide (132-135)
To a solution of hydrazide 131 in absolute alcohol (50 mL) was added different
arylisotliiocyanates and the reaction mixture refluxed for 5-8 h. After completion of
reaction monitored by TLC, the reaction mixture was cooled to RT and concentrated
under reduced pressure, the concentrated solution (25mL) was poured on crushed ice,
the precipitate so obtained was filtered off and washed with cold water, dried and
crystallized in ethanol to yield the pure thiosemicarbazides (132-135).
General procedure fo r the synthesis o f 3-mercapto~l,2,4-triazoles (136-139)
To a solution of thiosemicarbazide (132-135) in absolute alcohol (50 mL) was added
triethyl amine (ImL) and the reaction mixture refluxed for 5-8 h. After completion of
reaction monitored by TLC, the reaction mixture was cooled to RT and concentrated
under reduced pressure, the concentrated solution (25 mL) was poured on crushed ice,
the precipitate so obtained was filtered off, washed with cold water, dried and
crystallized in ethanol to yield the pure 3-mercapto-1,2,4-triazoles (136-139).
4-(3-chlorophenyl)-5-[{quinolin-8-yloxy}methyl]-4H-l,2,4~triazole-3-thiol (136)
Yield: 55 %; White crystals, m.p. 218-220 "C, Rf = 0.37 (CHCI3: MeOH; 9:1).
IR (KBr) cm -I : 3042, 2926, 1604, 1452, 1374, 1107
128
Chapter II, Section 4
‘H NMR (300 MHz, DMSO-dr,)
Maldi-MS (m/z)
Elenientai Analysis
Calculated
Found
: 8 5.12 (s, 2H, O-CH2), 7.21-7.60 (m, 8H, Ar-H),
8.35 (d, J - 8.4Hz, IH, Ar-H), 8.87 (d, J = 3.6
Hz, IH, Ar-H), 14.18 (s, IH, -SH).
: 368 (M^), 369 (M^+1).
; Calculated for molecular formula
C 18H13CIN4OS.
: C, 58.61; H, 3.55; N, 15.19.
: C, 58.24; H 3.46; N, 15.68%
4-(4-chlorophenyl)-5-[{quinolin-8-yloxy}methyl]-4H-l,2,4~triazole~3-thiol (137)
Yield: 65 %; White crystals, m.p. 230-232 "C, Rf = 0.34 (CHCI3: MeOH; 9:1).
IR (KBr) cm'' : 3038, 2926, 1603, 1452, 1374, 1109.
‘H NMR (300 MHz, DMSO-dg) : 5 5.13 (s, 2H, O-CH2), 6.98-7.54 (m, 7H, Ar-H),
8.33-8.36 (d, J == 9.3Hz, 2H, Ar-H), 8.87 (d, J
= 5.7Hz, IH, Ar-H), 14.14 (s, IH, -SH).
: 368 (M+), 369 (M ^ l) .
: Calculated for molecular formula
C|8H,3C1N40S.
: C, 58.61; H, 3.55; N, 15.19.
;C , 58.28; H, 3.49; N 15.45%
4-(4-methoxyphenyl)-5-[{quinoUn-8-yloxy}methyl]-4H-l,2,4-triazole~3-thiol (138)
Yield: 72 %; White crystals, m.p. 237-238 “C, Rf = 0.34 (CHCI3: MeOH; 9:1).
Maldi-MS (m/z)
Elemental Analysis
Calculated
Found
O-CH3
IR (KBr) cm-I 3042, 2922, 1601, 1450, 1371, 1108.
129
Chapter II, Section 4
'H NMR (300 MHz, DMSO-dfi)
Malcli-MS {m/7)
Elemental Analysis
Calcalated
Found
5 3.73 (s, 3H, O-CH3), 5.12 (s, 2 H, O-CH2), 6.89
(d, J - 8.7 Hz, 2H), 7.24 (d, ./== 7.2Hz, 2H, Ar-
H), 7.41-7.58 (m, 4H, Ar-H), 8.35 (d, 8.4
Hz, IH, Ar-H), 8.86 (d, 2.8Hz, IH, Ar- H),
14.10 (s, IH, -SH).
364 (M'‘), 365 (M^+1).
Calculated for molecular formula C |9H|f,N402S.
C, 62.62; H, 4.43; N, 15.37.
C, 62.34; H, 4.36; N, 15.70%
4-phenyl-5-[(quinolin-8~yloxy)methyl]-4H-l,2,4-triazole-3-thiol (141) Yield: 55%;
White crystals, m.p. 224-226 "C, Rf = 0.34 (CHCI3: MeOH; 9:1).
-IIR (KBr) cm
’H NMR (300 MHz, DMSO-dfi)
Maldi-MS {m/z)
Elemental Analysis
Calculated
Found
: 3056, 2927, 1605, 1454, 1376, 1107.
: 5 5.13 (s, 2H, O-CH2), 6.92-7.56 (m, 8H, Ar-H),
8.35 (d, J = 6.8 Hz, IH), 8.87-8.88 (d, J = 3.4
Hz, IH, Ar-H), 14.13 (s, IH, -SH).
334 (M"), 335 (M'^+1).
Calculated for molecular formula C 18H 14N4OS.
C, 64.65; H, 4.22; N, 16.75
C, 64.51; H, 4.19; N, 16.63%
General procedure for synthesis o f Quinoline based S-substituted 1,2,4-triazole
r'140-149;
To a solution of 3-mercapto-l,2,4-triazole (136-139, 0.5 mmol) in dry acetone (50
niL) was added different substituted phenacyl bromide (0.5 mmol), potassium
carbonate (1 mmol) and the reaction mixture refluxed for 4-8 h. After completion of
reaction monitored by TLC, the reaction mixture was filtered in hot condition,
concentrated under reduced pressure, the concentrated solution (25 mL) was poured
130
Chapter II, Section 4
on crushed ice, the precipitate so obtained was filtered, washed with cold water, dried
and crystallized in methanol to yield the pure compound (140-149).
2-[4~(3-chlorophenyl)-5-{(quinolin-8-yloxy)methyl}-4H-l,2,4-triazol-3-ylthioJ-l~(4-‘
chlorophenyl)ethanone (140) Yield: 54 %; White crystals, ra.p. 170 "C, Rf = 0.25
(CHCb: MeOH; 9:1).
,N.
,ci
NO
'Cl
IR (KBr) cm ' : 1677, 1587, 1557, 1486, 1455, 1375, 1302, 1112.
'H NMR (500 MHz, DMSO-dfi) : 8 4.96 (s, 2H, S-CH2), 5.21 (s, 2H, O-CH2), 7.01-
7.02 (d, 2H, 7.4 Hz), 7.17-7.19 (d, 2H, .7 -
6.7 Hz), 7.40-7.57 (m, 8H, Ar-H), 8.31-8.32 (s,
IH, ./= 3.6 Hz), 8.86-8,87 (d, IH, 3.4 Hz).
'■'’C-NMR (125 MHz, CDCI3)
Maldi-MS {m/z)
Elemental Analysis
Calculated
Found
: 8 41.42, 61.00, 111.98, 121.47, 121.92, 126.42,
127.22, 128.36, 129.05, 129.47, 130.47, 133.05,
134.41, 135.86, 139.77, 147.99, 149.41, 152.72,
168.65, 190.00.
: 520 (M+), 521 (M"+l).
: Calculated for molecular formula
C26H,8Cl2N402S.
: C 59.89, H 3.48, N 10.75.
: C 59.29, H 3.27, N 10.42%
2~[4-(3~chlorophenyl)-5-{(qumolin-8-yloxy)methyl}-4H-l,2,4-triazol-3-ylthio]-l-
(2,4-dihydroxyphenyl)ethanone (141) Yield; 62 %; White Crystals, m.p. 228-230
"C, R f - 0.32 (CHCI3: MeOH; 9:1).
"Cl
131
Chapter II, Section 4
-1IR (KBr) cm
‘H NMR (500 MHz, DMSO-dfi)
13C-NMR (125 MHz:, DMSO-de)
Maldl-MS {m/z) Elemental Analysis
Calculated
Found
: 3455, 3042, 2922, 1677, 1605, 1456, 1374, 1105.
: 5 4.91 (s, 2H, S-CH2), 5.18 (s, 2H, O-CH2), 6.86
(d, ./ = 8.6 Hz, 2H), 7.01-7.02 (d, J = 7.5 Hz,
IH), 7.29 (d, ./ = 6.6 Hz, IH), 7.41-7.49 (m,
4H), 7.52 (t, ./ = 4.5 Hz, 2H), 7.58 (d, ./ = 2.0
Hz, IH), 8.19-8.21 (d, y - 8.2 Hz,lH), 9.03-9.04
(d, 2.6 Hz, IH), 10.52 (s,lH, OH ), 10.61 (s,
IH, OH).
: 5 58.90, 63.90, 105.00, 121.35, 121.76, 122.10,
125.50, 126.50, 126.71, 126.76, 128.50, 128.80,
130.10, 130.30, 130.38, 134.79, 136.31, 146.36,
148.97,149.36,152.56,191.32.
: 518 (M' ), 520 (M‘ +2).
: Calculated for molecular formula
C26H,9C1N404S.
: C 60.17, H 3.69, N 10.80.
; C 60.67, H 3.44, N 11.02%
2-f4-(3-chlorophenyl)-5-{(quinolin-8-yloxy)methyl}~4H-lf2,4-triazol-3-ylthio]-l-(4-hydroxyphenyl)ethanone (142) Yield; 72 %; White Crystals, m,p. 218-220 "C, R f-
0.20 ( C H C I3 : MeOPI; 9:1).
'Cl
OH
.-IIR (KBr) cm'
‘H NMR (200 MHz, DMSO-df,)
13C-NMR (125 MHz, DMSO-de)
3450, 3046, 2926, 1674, 1601, 1450, 1371, 1108.
5 4.89 (s, 2H, S-CH2), 5.31 (s, 2H, O-CH2), 6.88
(d, 9.0Hz, 2H), 7.26 (d, 6.5Hz, IH), 7.82
(s, IH), 7.42-7.62 (m, 7H), 7.90 (d, J = 8.0 Hz,
2H), 8.88 (d, 2.6 Hz, IH), 10.53 (s, IH, OH).
5 42.34, 60.00, 111.00, 115.00, 121.00, 122.00,
126.00, 127.00, 129.00, 130.00, 131.00, 133.00,
132
Chapter II, Section 4
Makli-MS (m/z)
Elemental Analysis
Calculated
Found
135.00, 139.00, 149.00, 151.00, 152.00, 162.00,
191.00.
502 (M^), 503 (M '+l).
Calculated for molecular formula
C26H 19CIN4O3S.
C 62.09, H 3.81, N 11.14.
C 61.69, H 3.76, N 11.02%
2~[4-(3~chlorophenyl)S-{(qumolin-8-yloxy)methyl}-4H-l,2,4~triaz,ol~3-ylthio]-I~(4- bromophenyl)ethanone (143) Yield: 63 %; White Crystals, m.p.188-190 ”C; Rf =
0.22 (CHCI3: MeOH; 9:1).-Br
IR (KBr) cm -1
'H NMR (200 MHz, DMSO-dfi)
3046, 2926, 1674, 1601, 1450, 1371, 1108.
5 4.95 (s, 2H, S-CH2), 5.31 (s, 2H, O-CH2), 7.23
(d, J= 7.5Hz, 2H, Ar-H), 7.42-7.61 (m, 4H, Ar-
H), 7.76 (s, IH, Ar-H ), 7.78 (d, 7 - 8 .6Hz, 2H),
7.96 (d, ./= 8.5Hz, 2H, Ar-H), 8.32 (d, 4.7
Hz, 2H, Ar-H), 8.87 (s, 1H, Ar-H).
’^C-NMR (125 MHz, DMSO-de) ; 5 40.70, 61.28, 111.00, 121,00, 122.00, 126.62,
126.94, 127.84, 130.55, 130.87, 131.50, 132.36,
136.30, 149.86, 150.00, 151.00, 162.00, 190.00.
: 565 (M""), 566 (M'^+l).
; Calculated for molecular formula
C26H,8ClBrN402S.
; C 55.19, H 3.21, N 9.9.
: C 54.68, H 3.16, N 9.62%
Maidi-MS {m/z)
Elemental Analysis
Calculated
Found
133
Chapter II, Section 4
2-‘[4-(4-chiowphenyl)-5-{(quinolin-8-yiflxy)methyl}-4H-l,2,4-triazoi~3~yithioJ~I‘(4~
hydroxyphenyl)ethanone (14 4) Y ie ld ; 72 %; White Crystals, m .p. 180-182 Rf =
0.24 (CHCI3: MeOH; 9:1).
. N 'NS
Cl
I R ( K B r ) cm "’ : 3456, 3045, 2922, 1679, 1605, 1495, 1452, 1370,
1106.
'H NMR (200 MHz, DMSO-dfi) : 5 4.88 (s, 2H, S-CH2), 5.38 (s, 2H, O-CH2), 7.20
(d, J= 7.2 Hz, 2H), 7.43-7.55 (m, 3H), 7.63 (d,
J = 8.8Hz, 2H), 7.75 (d, J = 8.4Hz, 2H), 7.92
(d, J = 9.0 Hz, 2H), 8.29-8.31 (d, 2H, / = 4.4
Hz), 8 .81(d,lH ,J= 3.95Hz).
'-^C-NMR (125 MHz, DMSO-dg) : 5 48.5, 79.00, 111.00, 122, 126.42, 129.06,
129.48, 130.32, 131.80, 139.76, 149.65, 150.95,
153.30, 192.34.
Maldi-MS {m/z) : 502 (M‘"), 503 (M'*'+l).
Elemental Analysis : Calculated for molecular formula
C26H19CIN4O3S.
Calculated : C 62.09, H 3.81, N 11.14.
Found : C 61.69, H 3.76, N 11.02%
2-[4~(4-chlorophenyl)-5-{(quinolin-8-yloxy)methyl}-4H-l,2,4~triazol-3-ylthioj-l~(4-bromophenyl)ethanone (145) Yield: 68 %; White Crystals, m.p. 168-170 ”C; Rf =
0.28 (CHCI3: MeOH; 9:1).
IR (KBr) cm ’
"H NMR (200 MHz, DMSO-ds)
-Br
; 3045, 2926, 1681, 1601, 1495, 1455, 1370, 1104
: 8 4.91 (s, 2H, S-CH2), 5.28 (s, 2H, O-CH2), 7.20
(d, 2H, 7.2 Hz), 7.43-7.55(m, 3H), 7.63 (d.
134
Chapter II, Section 4
2H, J = 8.8 Hz), 7.75 (d, 2H, J = 8.4Hz), 7.92
(d, 2H, J = 9.0 Hz), 8.30 (d, 2H, J - 4.4 Hz),
8.83 (d, IH, J - 3 .9 Hz).
‘-^C-NMR (125 MHz, DMSO-dfi) ; 5 44.5, 79.00, 112 .00 , 1 2 1 .0, 128.00, 126.42,
128.48, 129.00, 131.32, 132.80, 137.72, 148.65,
151.95, 152.32, 191.34.
MALDI MS {m/z) : 565 (M"''), 566 (M ^ l) .
Elemental Analysis : Calculated for molecular formula
C26Hi8ClBrN402S.
Calculated ; C 62.09, H 3.81, N 11.14.
Found : C 61.69, H 3.76, N 11.06%
2-f4-(4-methoxyphenyl)~S-((quinolm-8~yloxy)methyl}~4H~I,2,4-tria7.ol~3-ylthio]-I-
phenylethanone (146) Yield: 49 %; White Crystals, m.p.172-174 "C, Rf = 0.31
(CHCI3: MeOH;9:l).
IR (KBr) cm'*
'H NMR (200 MHz, DMSO-de)
'■^C-NMR (125 MHz, DMSO-d<;)
MALDI MS (wA)
Elemental Analysis
Calculated
: 3042, 2927, 1678, 1515, 1445, 1408, 1379, 1262,
1111.: 5 3.72 (s, 3H, O-CH3 ), 4.86 (s, 2H, S-CH2),4.98
(s, 2H, O-CH2), 6.81 (d, 8.8 Hz, 2H), 7,24 (d,
J = 7.3Hz, IH), 7.36 (d, J = 7.3 Hz, 2H), 7.46-
7.59 (m, 7H), 8.33(d, J = 6.2 Hz, 2H), 8.87 (d, J = 4.1Hz, IH).
: 5 40.50, 56.00, 79.00, 114.14, 121.12, 126.42,
128.00, 129.00, 128.48, 131.32, 132.80, 137.72,
148.65,151.95,152.32,191.38.: 482 (M^), 483 (M'^+l).
: Calculated for molecular formula
C27H22CIN4O3S.
: C 67 ,2 ,H 4.6 ,N 11.61.
135
Chapter //, Section 4
Found : C 67.69, H 4.46, N 11.52%
2~(4-phenyl-5-[(qmnolm-8~yloxy)methyl]~4H-l,2,4~triaz,ol~3~ylthio)-l-
phenyletkamme (147) Yield: 62 %; White Crystals, m.p.160-162 ”C, Rf = 0.25 {n-
liexane : ethylacetate; 4:6).
IR (KBr) cm-1
'H NMR (200 MHz, DMSO-dg)
‘'^C-NMR (125 MHz, DMSO-de)
MALDI MS {m/z)
Elemental Analysis
Calculated
Found
-N
N'-n0 ' ^ X
3450, 3046, 2926, 1674, 1601, 1450, 1371, 1108.
5 5.02 (s, 2H, S-CH2), 5.32 (s, 2H, 0-CH2),7-26-
7.28 (d, J = 7.36 Hz, 2H), 7.47-7.70 (m, 9H),
7.69 (d, J = 8.3 Hz, 2H), 8.00 (d, J = 8.30 Hz,
2H), 8.37 (d, J = 7.80 Hz,lH), 8.93(1H, J= 2.65
Hz).
8 42.50, 57.00, 78.00, 121.00, 126.42, 128.00,
128.48, 129.00, 130.32, 132.80, 138.72, 149.65,
151.95, 152.82, 191.34.
452 (M-"), 453 (M'^+1).
Calculated for molecular formula C26H20N4O2S
C 69.01, H 4.45, N 12.38.
C 68.69, H 4.52, N 12.18%
2-[4-(4-methoxyphenyl)~5-{(quinolin-8-yloxy)methyl}-4H-l,2,4-triazol-3-ylthio]-l-
(4~hydroxyphenyl)ethanone (148) Yield: 68 %; White Crystals, m.p. 230-232 "C, Rf
- 0.18 (CHCI3 : MeOH; 9:1),
N-n9 ^ ^ ^ X
IR (KBr) cm’’
'H NMR (200 MHz, DMSO-df,)
; 3450,3046, 2926, 1674, 1601, 1450, 1371, 1108.
; 5 3.74 (s, 3H, O-CH3), 4.88 (s, 2H, S-CH2), 5.25
(s, 2H, O-CH2), 6.88 (d, / = 8.60 Hz, 2H), 6.96
(d, J - 8.80 Hz, 2H), 7.24 (d, J - 6.80 Hz, IH),
136
Chapter II, Section 4
''^C-NMR (125 MHz, DMSO-dr.)
MALDI MS {m/z)i Elemental Analysis
Calculated
Found
7.42-7.89 (m, 5H), 7.92 (d, J = 8.65 Hz, 2H),
8.34 (d, 8.1 Hz, IH), 8 .8 8 (d, J - 3.00 Hz,
IH).
; 5 34.46, 56.54, 78.00, 111.00, 114.00, 121.00,
125.42, 129.06, 129.48, 130.32, 131.80, 139.76,
149.65, 150.95, 155.30, 159.00, 162.00, 191.00.
498 (M ), 499 (M ^l).
: Calculated for molecular formula C27H22N4O4S.: C 65.05, H 4.45, N 11.24.
: C 65.63, H 4.66, N 11.12%
2-f4-(4-methoxyphenyl)-5-{(quinolin~8-yloxy)methyl}-4H~l,2,4-triazol-3-ylthio)-l- (4-bromophenyl)ethanone (149) Yield: 74 %; White Crystals, m.p. 158-160 "C, Rf =
0.25 (CHCI3: MeOH; 9:1),N''N
-IIR (KBr) cm
'H NMR (200 MHz, DMSO-dfi)
13C-NMR (125 MHz, DMSO-ds)
MALDI MS {m/z)
Elemental Analysis
Calculated
Found
CH3
: 3056, 2926, 1674, 1601, 1450, 1371, 1108.
: 8 3.79 (s, 3H, O-CH3), 4.96 (s, 2H, S-CH2), 5.27
(s, 2H, O-CH2), 6.85 (d, ./= 8 .6 Hz, 2 H), 7.23
(d, J = 6 .6 Hz, IH), 7.41-7.65 (m, 6 H), 7.79 (d,/
= 8.4 Hz, 2H), 7.97 (d, 7 = 8.5 Hz, 2H), 8.30 (d,
1.3 Hz, IH).
; 5 42.50, 55.43, 78.00, 114.00, 120.00, 126.42,
128.00, 128.48, 131.32, 133.00, 137.72, 148.65,
150.00, 152.32, 193.34.
: 561 (M""), 562 (M^+1).
: Calculated for molecular formula
C 2 7 H 2 iC lB r N 4 0 3 S .
; C 57.76, H 3.77, N 9.98.
: C 57,24, H 3.68, N 10.04%
137
Chapter II, Section 4
2=4.4.2.2. Biotoglcal activity
2.4.4.2.2.1. A nti~inflammatorv activityAll the synthesized compounds were evaluated for their anti-inflammatory activity against carrageenan-induced rat paw oedema in Albino Wistar rats weighing 150-200 g. The standard dmg Indom ethacin was used at dose of 10 mg/kg body weight and test drugs were used equimolar to the standard dmg. The method used was the same as discussed in section-1 of this chapter. The results of anti- inflammatory activity are presented in table < .2 and figure 4.1.
2.4.4.2.2.2. Analeesic activityThe analgesic activity was determined using tail flick method in Albino Wistar rats weighing 150-200 g. Indomethacin (10 mg/kg body weight) was used as standard dmg and test drugs were used equimolar to the standard drags. The same method was used as given in section-1 of this chapter. The results are presented in table 4.3 and
figure 4.2.
2.4.4.2.2.3. Histopatholocical studiesFor the h istopathological study, same m ethods were used as given in section-1 of this chapter. The dmgs were used at doses three times to the doses used for antiinflammatory activity. The results are presented in table 4.4 and figmre 4.3,
2.4.4.2.2.4. Antimicrobial activityAll the synthesized compounds were evaluated for their antimicrobial activity against various bacterial and fungal strains. Same methods were used as given in section-1 of this chapter. The results are presented in table 4.5.
138
Chapter II, Section 4
2.4.43o ConclusionIn conclusion, we have synthesized a small library of ten compounds 140-149 successfully using the standard methodology. The newly synthesized compounds were evaluated for their anti-inflammatory, analgesic, ulcerogenic and antimicrobial studies. Compound 149 has exhibited potent anti-inflammatory, comparable analgesic activity with no ulcer. Other members 142 and 143 from this series also exhibited significant anti-inflammatory activity with reduced ulcer. Compound 149 may be considered as promising candidate for development of new anti-inflammatory agent. From antimicrobial screening it was concluded that most of the compounds from this series showed moderate antimicrobial activity.
139
c :
■I i i c C
r '
..134 .
*1.30*
| |a t » | t t | | I t » |' :i .. ■: | V
I
8 i« » » « |« 8 « |n || |f |f l | | | :
iMiiiii s.s.Jij IksJ iilg iiJ il.ih lld} -.1| | g |» | ; » I i I S i m I|M| I
'H NMR spectrum of compound 149
' C NMR spectrum of compound 149
f I
*1
i
mmm
Mass spectrum of compound 149
IR spectrum of compound 149
Chapter II, Section 4
2,5. References
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A. and Meral, O., Bioorg. Med. Chem., 2007, 15, 5738-5751.
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[10] Shi, Y. and Zhau, C., Bioorg. Med. Chem. Lett., 2011, 21, 956-960.
[11] Guan, L., Sun, X. Y., Tian, G. R., Chi, K. Y. and Quan, Z. S., Turk J. Chem..,
2008,32, 181-189.
[12] Tomasz, P., Monika, W., Agata, S., Kosikowska, U. and Anna, M., Eur. J. Med.
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1808-1814.
[14] Navidpour, L., Hameed, S., Abdi, K., Amini, M,, Ghahremani, M. H,,
Dehpourd, A. R. and Abbas, S., Bioorg. Med. Chem., 2006, 14, 2507-2517.
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