chapter ii synthesis of 1,2,4-triazole derivatives and...

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


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


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


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


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


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


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


mihr U5hr

Figure 1.1. Anti-inflammatory activity of biphenyl based 1,2,4 triazole derivatives.

31


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


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


-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


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


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


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


-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


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


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).


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


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


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


• 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-'


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


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-'


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


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"


: 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


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


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


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



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).


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


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


; 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


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).


C3oH24Bi'N303S.

: C, 61.44; H, 4.12; N, 7.16.

:C , 61.48; H, 4.09; N, 7.13%.

53


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'


'•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


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).


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-


IR (ICBr) cm‘‘

NMR (400 M Hz, CDCh)


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"


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



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)


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~


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).


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 =


CHi

59


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).


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'



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


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*



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).


C28H21FN4O3S.

: C, 65.61; H, 4.13; N, 10.93.

: C, 65.67; H, 4.09; N, 10.90%.

61


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)


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



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'*



Maldi-M S {m/z:)


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).


C29H2iClBrN302S.

: C, 58.94; H, 3.58; N, 7.11.

: C, 59.02 H, 3.59; N, 7.08%.

63


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 =


‘-■’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 =


Maldi-M S {m/z)

Elem ental Analysis

Calculated

Found

'CH,

IR (KBr) cm' : 2912, 1681, 1607, 1093.

64



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).


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-


IR (KBr) cm '

'H NM R (400 MHz, CDCI3)

Elemental Analysis

Calculated

Found

: 2924, 1680, 1584, 1107.

: 5 4.51 (s^+1).


C29H22CIN3O2S.

:C , 68.03; H, 4.33; N, 8.21.

: C, 68.09; H, 4.30; N, 8.19%.

65


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'*


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


-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"



: 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


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).


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)


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).


C, 65.20; H, 4.38; N, 10.14

C, 65.22; H, 4.37; N, 10.12%.

6 8


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”


‘- 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).


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 =


//^"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


'•"'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).


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'



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).


C, 70.22; H, 4.38; N, 9.36.

C, 70.14; H, 4.35; N, 9.34%.

70


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 («-


NO2

IR (KBr) cm’*


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 '


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



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


NM R (75 M Hz, CDCI3)

ES-MS (m/z)


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"'


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).


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



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'


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


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


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


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


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


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


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


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


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


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


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


% 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


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


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


. 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


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


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


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


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-


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


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


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 '‘


’•'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


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"



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).


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


IR (KBr) cm"*

‘H NM R (3§(S M Hz, 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).


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 '


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


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)


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’’


'^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).


C32H28 CIN5O4S.

96


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'


‘'"’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)


CjiHaaClNsOsS.

: C, 61.00; H, 4.48; N, 11.11

: C 61.04; H 4.47; N 11.12%.

97


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)


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).


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


IR (KBr) cm-’

'H NM R (300 M Hz, 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 )


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


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'



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).


C30H23CIN6O4S.

: C, 60.15; H, 3.87; N, 14.03

; C 60.19; H 3.85; N 14.04%.

100


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


‘^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


IR (KJBr) cn f'

'H NMR (300 M Hz, CDCb)


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).


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'



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


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).


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 ,


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


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


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 ^


Chapter II

Section 3

Synthesis of 3-mercapto 1,2,4-triazole derived

mannich bases and their Biological Activities


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


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


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.


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


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


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


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


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


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)


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


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'



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


'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).


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).


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


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


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)


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.


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


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


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^ ‘

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I S ,

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m ■ tti: ■P><{-.M.'Jki C4 i • •‘-1-;3

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'H NMR spectrum of compound 124

MOO

©3O

<3?O

o

MO

oo

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o

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128.34127.03126.76IIJ.ll

. 77,43 77.00 7S.38 -— 6')M ■•—'-<>6,8!

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> <j> Q O ih-*; )-* nip 0V«? 0 KC3iCn::ii'

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‘ ■C NMR spectrum of compound 124

S3JL ^

8

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Chapter II

Section 4

Synthesis of Quinoline based 1,2,4-triazole

derivatives and their Biological Activities


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


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


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


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].


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


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


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


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"“


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


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


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


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


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


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



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


‘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).


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) .


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


'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


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).


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


-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).


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


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).


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


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).


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


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


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).


C27H22CIN4O3S.

: C 67 ,2 ,H 4.6 ,N 11.61.

135


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


''^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


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).


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


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


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



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142

chapter ii synthesis of 1,2,4-triazole derivatives and...

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