Synthesis and biological evaluation of thiazoline derivatives as new antimicrobial and anticancer agents

European Journal of Medicinal Chemistry 74 (2014) 264e277

Contents lists available at ScienceDirect

European Journal of Medicinal Chemistry

journal homepage: h ttp://www.elsevier.com/locate/ejmech

Original article

Synthesis and biological evaluation of thiazoline derivatives as new

antimicrobial and anticancer agents

,

b

c

Mehlika Dilek Altıntop ^a, Zafer Asım Kaplancıklı ^a, Güls¸ en Akalın Çiftçi , Rasime Demirel

*

^aAnadolu University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, 26470 Eskis¸ehir, Turkey

^bAnadolu University, Faculty of Pharmacy, Department of Biochemistry, 26470 Eskis¸ehir, Turkey

^cAnadolu University, Faculty of Science, Department of Biology, 26470 Eskis¸ehir, Turkey

a r t i c l e i n f o

a b s t r a c t

N⁰-(3,4-Diarylthiazol-2(3H)-ylidene)-2-(arylthio)acetohydrazides were synthesized and evaluated for

their antimicrobial activity and cytotoxicity against NIH/3T3 cells. Compound 22 bearing 1-phenyl-1H-

tetrazole and p-chlorophenyl moieties was found to be the most promising antibacterial agent against

Pseudomonas aeruginosa, whereas compound 23 bearing 1-phenyl-1H-tetrazole and p-bromophenyl

moieties was the most promising antifungal agent against Candida albicans. The most effective de-

rivatives were also evaluated for their cytotoxicity against C6 glioma cells. The results indicated that

Article history:

Received 10 August 2013

Received in revised form

30 December 2013

Accepted 30 December 2013

Available online 10 January 2014

compound 17 bearing 1-phenyl-1H-tetrazole and nonsubstituted phenyl moieties (IC₅₀¼ 8.3 ꢀ 2.6

mL) was more effective than cisplatin (IC₅₀¼ 13.7 ꢀ 1.2 g/mL) against C6 glioma cells. Compound 17

also exhibited DNA synthesis inhibitory activity on C6 cells. Furthermore, compound 17 showed low

toxicity to NIH/3T3 cells (IC₅₀¼ 416.7 ꢀ 28.9 g/mL).

mg/

Keywords:

Thiazoline

Hydrazone

Antimicrobial activity

Cytotoxicity

m

DNA synthesis inhibitory activity

1. Introduction

of interest as privileged scaffolds due to their synthetic and bio-

logical importance. Compounds bearing thiazoline moiety have

been reported to exhibit a wide spectrum of biological effects

including antimicrobial activity [11e18]. In addition, hydrazidese

hydrazones have received considerable attention as important

pharmacophores in medicinal chemistry [19,20]. Isoniazid, which

possesses a hydrazide moiety in its molecular structure, is the

frontline drug employed in the treatment of tuberculosis [21].

Nifuroxazide, which is a nitrofuran antibacterial agent bearing a

hydrazone moiety, is widely used as an intestinal antiseptic. Many

studies have also conﬁrmed that hydrazone derivatives of isoniazid

and other hydrazides exhibit signiﬁcant antimicrobial activity [19e

26].

Infectious diseases caused by bacteria and fungi have emerged

as important causes of morbidity and mortality worldwide due to

their ability to thwart therapeutic regimens by rapidly evolving

resistance to antimicrobial agents [1e6].

Eukaryotic pathogens such as fungi pose a particular therapeutic

challenge since they share a close evolutionary relationship with

their human hosts, limiting the number of drug targets that can be

exploited to selectively kill the pathogen [6]. The treatment of

fungal infections, particularly those caused by drug-resistant fungal

pathogens is often complicated by high toxicity, low tolerability, or

narrow spectrum of activity [3e7].

In the last few decades, the greatly increased incidence of life-

threatening bacterial and fungal infections has resulted in a cor-

responding increase in demand for new effective antimicrobial

agents which inhibit the growth of pathogens or kill them and have

no or least toxicity to host cells [1e8].

1-Substituted-1H-tetrazole-5-thiol and 5-methyl-1,3,4-thiadia

zole-2-thiol have gained great importance in the synthesis of

pharmacologically active drugs. Some synthetic

b-lactam antibi-

otics possess 5-thio-1-methyl-1H-tetrazole (MTT) or thio-linked

thiadiazole as the side chain [10,27].

Thiazoles are found in many biologically active compounds,

including natural products and pharmaceutical agents [9,10]. The

reduced forms of thiazoles, thiazolines, have attracted a great deal

Triazole antifungal agents, play a leading role in the treatment of

systemic fungal infections due to their broad spectrum and

improved safety proﬁle. Fluconazole, itraconazole, voriconazole,

and posaconazole are widely used antifungal drugs bearing triazole

ring for the treatment of systemic fungal infections [28,29].

Medicinal chemists have also carried out considerable research

* Corresponding author. Tel.: þ90 222 3350580/3776; fax: þ90 222 3350750.

E-mail address: zakaplan@anadolu.edu.tr (Z.A. Kaplancıklı).

for novel antimicrobial agents bearing

a pyrimidine moiety.

http://dx.doi.org/10.1016/j.ejmech.2013.12.060

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

265

ClCH₂COOC₂H₅

O

NH₂NH₂. H₂O

Ar SH

S

K₂CO₃

OC₂H₅

Ar

NHNH₂

Ar

B1-6

A1-6

N

S

C

O

R

Br

O

H

N

H

N

S

N

S

R

H

N

Ar

N

Ar

S

N

H

S

O

C1-6

1-40

Scheme 1. The synthetic route for the preparation of the target compounds (1e40).

Trimethoprim and sulfadiazine are chemotherapeutic drugs con-

taining a pyrimidine moiety currently used in the treatment of

infectious diseases [10,30].

On the basis of these ﬁndings, herein we reported the synthesis of

a series of thiazoline derivatives bearing a hydrazone moiety and

focusedontheirpotentialantimicrobial effects and cytotoxicity. Thio-

linked triazole, tetrazole, thiadiazole and pyrimidine were preferred

as the side chain while designing the molecules. Furthermore, all

compounds were evaluated for their cytotoxic effects against NIH/

3T3 cell lines. The most effective antimicrobial derivatives were also

Table 1

Some properties of the compounds (1e40).

Compound

R

Ar

Yield (%)

M.p. (^ꢁC)

Molecular formula

Molecular weight

1

2

3

4

5

6

7

8

H

4-Methyl-4H-1,2,4-triazol-3-yl

1-Methyl-1H-tetrazol-5-yl

1-Phenyl-1H-tetrazol-5-yl

5-Methyl-1,3,4-thiadiazol-2-yl

Pyrimidin-2-yl

70

80

70

71

72

75

76

81

75

80

74

72

75

77

78

81

78

85

75

73

75

83

84

86

79

88

76

75

78

85

86

89

80

89

77

75

79

87

88

90

123e126

100e103

109e112

148e151

167e169

260e262

260e263

246e247

202e206

78e80

224e225

124e125

218e219

223e227

229e230

244e245

71e74

C₂₀H₁₈N₆OS₂

C₂₀H₁₇N₇O₃S₂

C₂₁H₂₀N₆OS₂

C₂₁H₂₀N₆O₂S₂

422

467

436

452

440

456

501

447

423

468

437

453

441

457

502

448

485

530

499

515

503

519

564

510

439

484

453

469

457

473

518

464

419

464

433

449

437

453

498

444

NO₂

CH₃

OCH₃

F

Cl

Br

CN

H

NO₂

CH₃

OCH₃

F

C

₂₀H₁₇FN₆OS₂

₂₀H₁₇ClN₆OS₂

₂₀H₁₇BrN₆OS₂

₂₁H₁₇N₇OS₂

₁₉H₁₇N₇OS₂

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

C₁₉H₁₆N₈O₃S₂

C₂₀H₁₉N₇OS₂

C₂₀H₁₉N₇O₂S₂

C

₁₉H₁₆FN₇OS₂

₁₉H₁₆ClN₇OS₂

₁₉H₁₆BrN₇OS₂

₂₀H₁₆N₈OS₂

₂₄H₁₉N₇OS₂

Cl

Br

CN

H

NO₂

CH₃

OCH₃

F

235e237

232e233

230e231

237

C₂₄H₁₈N₈O₃S₂

C₂₅H₂₁N₇OS₂

C₂₅H₂₁N₇O₂S₂

C

₂₄H₁₈FN₇OS₂

₂₄H₁₈ClN₇OS₂

₂₄H₁₈BrN₇OS₂

₂₅H₁₈N₈OS₂

₂₀H₁₇N₅OS₃

Cl

Br

246e248

242e243

233e234

156e160

250e252

240e241

248e249

228e231

244e245

237e239

254e255

77e78

CN

H

NO₂

CH₃

OCH₃

F

C₂₀H₁₆N₆O₃S₃

C₂₁H₁₉N₅OS₃

C₂₁H₁₉N₅O₂S₃

C

₂₀H₁₆FN₅OS₃

₂₀H₁₆ClN₅OS₃

₂₀H₁₆BrN₅OS₃

₂₁H₁₆N₆OS₃

₂₁H₁₇N₅OS₂

Cl

Br

CN

H

NO₂

CH₃

OCH₃

F

Cl

Br

CN

Pyrimidin-2-yl

260e263

84e87

C₂₁H₁₆N₆O₃S₂

C₂₂H₁₉N₅OS₂

C₂₂H₁₉N₅O₂S₂

218e219

241e242

247e248

245e246

256e258

C

₂₁H₁₆FN₅OS₂

₂₁H₁₆ClN₅OS₂

₂₁H₁₆BrN₅OS₂

₂₂H₁₆N₆OS₂

266

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

investigated for their cytotoxicity against C6 rat glioma cells. Among

these derivatives, the most effective anticancer compounds were

evaluated for their DNA synthesis inhibitory activity.

exhibited more signiﬁcant antimicrobial activity against fungi than

bacteria.

Among bacteria species, Pseudomonas aeruginosa was the most

susceptible bacterium to compounds 5, 22 and 40. These com-

pounds and streptomycin exhibited the same level of antibacterial

2. Chemistry

activity against P. aeruginosa with a MIC value of 125 mg/mL. Among

The synthesis of the thiazoline derivatives (1e40) was carried

out according to the steps shown in Scheme 1. In the initial step,

ethyl 2-[(aryl)thio]acetate derivatives (A1e6) were synthesized via

the reaction of thiols with ethyl chloroacetate in the presence of

potassium carbonate. The treatment of the ester derivatives (A1e6)

with hydrazine hydrate afforded 2-[(aryl)thio]acetohydrazides

(B1e6) [31e34]. 1-(Arylthioacetyl)-4-phenyl thiosemicarbazides

(C1e6) were obtained by the reaction of the hydrazides (B1e6)

with phenyl isothiocyanate [34e36]. In the last step, the ring

closure reaction of the thiosemicarbazide derivatives (C1e6) with

2-bromoacetophenone/4⁰-substituted-2-bromoacetophenone gave

the target compounds (1e40) [18,36]. Some properties of the

compounds were given in Table 1.

these derivatives, compound 22 bearing 1-phenyl-1H-tetrazole and

p-chlorophenyl moieties can be identiﬁed as the most promising

antibacterial agent against P. aeruginosa due to its inhibitory effect

on P. aeruginosa and low toxicity to NIH/3T3 cells

(IC₅₀¼ 313.3 ꢀ 15.3

mg/mL).

Compounds 3 and 23 were more effective than ketoconazole,

whereas other derivatives and ketoconazole exhibited the same

level of antifungal activity against Candida albicans. Compounds 3

and 23 exhibited the inhibitory activityagainst C. albicans with a MIC

value of 125

activity with a MIC value of 250

compounds 3 and 23 were determined as 130.0 ꢀ 20.0

g/mL, respectively. According to MTTassay, compound

m

g/mL, whereas ketoconazole exhibited the inhibitory

g/mL. In addition, IC₅₀values of

g/mL and

m

433.3 ꢀ 28.9

m

23 bearing 1-phenyl-1H-tetrazole and p-bromophenyl moieties

seems to be the most promising anticandidal agent owing to its

inhibitory effect on C. albicans and low toxicity to NIH/3T3 cells.

Compounds 1e40 exhibited the inhibitory activity against

3. Results and discussion

The structures of new compounds were conﬁrmed by FT-IR, ¹H

NMR, ¹³C NMR, mass spectral data, and elemental analysis.

Aspergillus parasiticus with a MIC value of 250

mg/mL, whilst

In the IR spectra of C-2 and C-5, the C]O stretching vibrations

gave rise to a band at 1700e1650 cm^ꢂ1. The stretching bands for Ne

H group were observed in the region 3400e3200 cm^ꢂ1. The

stretching bands for aromatic and aliphatic CeH groups occurred at

3100e3000 cm^ꢂ1and 2980e2900 cm^ꢂ1, respectively. C]N, C]C

stretching and NeH bending vibrations were observed in the re-

Table 2

Antibacterial activity of the compounds (1e40) as MIC values (mg/mL).

Compound

A

B

C

D

E

F

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

250

125

250

500

250

500

250

500

250

500

250

125

250

125

250

125

250

500

125

250

125

250

125

250

125

250

125

250

125

250

125

250

125

250

500

250

125

250

125

250

125

250

125

250

500

250

500

250

500

250

500

250

500

250

500

250

500

250

500

250

500

250

gion 1600e1450 cm^ꢂ1

.

In the IR spectra of compounds 1e40, the compounds had a

strong, characteristic band in the region 1750e1650 cm^ꢂ1due to

the C]O stretching vibration. The stretching bands for NeH group

occurred at 3500e3200 cm^ꢂ1. The aromatic and aliphatic CeH

stretching vibrations gave rise to bands at 3120e3000 cm^ꢂ1and

2990e2700 cm^ꢂ1, respectively. C]N, C]C stretching and NeH

bending vibrations were observed in the region 1620e1450 cm^ꢂ1

In the IR spectra of the compounds bearing cyano substituent, the

.

stretching bands for C^N group occurred at 2250e2200 cm^ꢂ1

.

In the ¹H NMR spectra of C-2 and C-5, the signal due to the

thiosemicarbazide protons appeared in the region 9.5e10.5 ppm.

The signal due to the SeCH₂protons was observed in the region

3.99e4.11 ppm.

In the ¹H NMR spectra of compounds 1e40, the signal due to

the hydrazone proton appeared in the region 10e13 ppm. The

signal due to the SeCH₂protons was observed in the region 3.86e

4.37 ppm. In the ¹H NMR spectra of some compounds, NeH and

SeCH₂protons gave rise to two singlet peaks in accordance with

the presence of the E and Z isomers [37,38]. The signal due to the

thiazoline proton was observed in the region 6.5e7.3 ppm. Other

aromatic and aliphatic protons were observed at expected

regions.

In the ¹³C NMR spectra of all compounds, the signal due to the

SeCH₂carbon was observed in the region 32e36 ppm.

In the ¹³C NMR spectra of C-2 and C-5, the signal due to the C]S

carbon was observed at 181.68 and 181.52 ppm, respectively. The

signal due to the C]O carbon was observed at 167e168 ppm.

In the ¹³C NMR spectra of compounds 1e40, the signal due to

the C]O carbon was observed at 160e180 ppm. Other aromatic

and aliphatic carbons were observed at expected regions.

The mass spectral data were also consistent with the assigned

structures. All compounds gave satisfactory elemental analysis.

Compounds 1e40 were tested in vitro against pathogenic bac-

teria and fungi strains. As shown in Tables 2 and 3, the compounds

33

34

35

36

37

38

39

40

Streptomycin

31.25

7.81

31.25

15.625

A: S. aureus (NRRL B-767), B: L. monocytogenes (ATCC-7644), C: E. coli (ATCC-25922),

D: P. aeruginosa (ATCC-254992), E: M. luteus (NRLL B-4375), F: B. subtilis (NRS-744).

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

267

Table 3

of compounds 1, 3, 8, 9, 10, 11, 16, 17, 19, 21, 22, 23, 24, 25, 27, 28, 29,

30, 35, 36, 37, 38, 39 against C6 cells for 24 h were presented in

Tables 4 and 5, respectively. Compound 17 bearing 1-phenyl-1H-

tetrazole and nonsubstituted phenyl moieties can be identiﬁed as

the most promising anticancer agent against C6 glioma cell lines

due to its inhibitory effect on C6 cells and low toxicity to NIH/3T3

Antifungal activity of the compounds (1e40) as MIC values (mg/mL).

Compound

A

B

C

D

E

F

G

H

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

250

125

250

125

250

125

250

125

250

125

250

125

250

125

250

500

250

500

250

125

250

125

250

125

250

125

250

500

250

125

250

125

250

cells (IC₅₀¼ 416.7 ꢀ 28.9

inhibitory effect on C6 cells with an IC₅₀value of 8.3 ꢀ 2.6

whereas cisplatin showed its anticancer activity with an IC₅₀value

of 13.7 ꢀ 1.2 g/mL.

mg/mL). Compound 17 exhibited its

mg/mL,

m

The immunostaining procedure was carried out with speciﬁc

anti-BrdU antibodies in the S-phase of the cell cycle. DNA synthesis

inhibitory activity of compounds 16, 17, 19, 21, 23, 27, 28, 29 and 30

which exhibited signiﬁcant cytotoxic activity in MTT assay on C6

cell lines is presented in Fig. 1. DNA synthesis inhibition activity of

the compounds was evaluated for 24 h. C6 cell lines were incubated

with three different concentrations of the compounds that were

determined according to their IC₅₀values. Cisplatin was used as a

positive control. The tested compounds showed inhibitory activity

on dose dependent manner. These results show that the com-

pounds tested in this assay cause DNA synthesis inhibition. Among

these compounds, the most potent inhibitors of DNA synthesis

against C6 cell lines were found as compounds 16, 17 and 19. DNA

synthesis inhibition caused by the concentration of IC₅₀for com-

pounds 16, 17 and 19 tested in C6 cell line were similar. Although

compounds 16, 17 and 19 had similar inhibitory effects on DNA

synthesis activity, compound 17 was the best among them due to

low IC₅₀value against C6 glioma cells (Table 5). On the other hand,

DNA synthesis inhibition caused by the concentration of IC₅₀for

compounds 21, 23, 27, 28, 29 and 30 were similar.

33

34

35

36

37

38

39

40

All compounds were also evaluated for their cytotoxic effects

against NIH/3T3 cell lines. The MTT assay indicated that compounds

32 and 40 possessed the highest cytotoxicity (IC₅₀< 3.9

mg/mL),

whereas compounds 19, 35 and 37 exhibited the lowest cytotox-

icity (IC₅₀> 500

mg/mL) against NIH/3T3 cells among the com-

pounds (Table 5).

Ketoconazole

7.8

4. Conclusion

A: A. parasiticus (NRRL 465), B: P. chrysogenum (NRRL 1951), C: T. harzianum (NRRL

20565), D: A. ochraceus (NRRL 3174), E: F. solani (NRRL-13414) F: F. moniliforme

(NRLL 1866), G: F. culmorum (wild culture), H: C. albicans (ATCC-22019).

In the present paper, we described the synthesis of thiazoline

derivatives bearing a hydrazone moiety, which were tested in vitro

against pathogenic bacteria and fungi.

ketoconazole exhibited the inhibitory activity with a MIC value of

125

same level of antifungal activity against Aspergillus ochraceus with a

MIC value of 250 g/mL.

m

g/mL. Compounds 1e40 and ketoconazole also showed the

The microbiological results revealed that the compounds

exhibited more signiﬁcant antimicrobial activity against fungi than

bacteria.

m

Compounds 1e40 were more effective than ketoconazole

against Fusarium solani and Fusarium moniliforme. These com-

pounds exhibited the inhibitory activity against F. solani and

Compound 22 bearing 1-phenyl-1H-tetrazole and p-chlor-

ophenyl moieties can be identiﬁed as the most promising anti-

bacterial agent against P. aeruginosa due to its inhibitory effect

on P. aeruginosa and low toxicity to NIH3T3 cells

F. moniliforme with a MIC value of 250

mg/mL, whereas ketocona-

zole exhibited the inhibitory activity with a MIC value of 500

m

g/mL.

(IC₅₀¼ 313.3 ꢀ 15.3

mg/mL), whereas compound 23 bearing 1-

The microbiological results demonstrated that the antifungal ef-

fects of the compounds on A. parasiticus, A. ochraceus, F. solani and

F. moniliforme did not depend on the substituents.

phenyl-1H-tetrazole and p-bromophenyl moieties was found to

be the most promising anticandidal agent owing to its inhibitory

effect on C. albicans and low toxicity to NIH3T3 cells

Compounds 1e40 were found to be more potent than ketoco-

nazole against Fusarium culmorum. Compounds 11, 15, 17, 19, 20 and

(IC₅₀¼ 433.3 ꢀ 28.9

mg/mL). Furthermore, all compounds exhibited

signiﬁcant antifungal activity against T. harzianum, A. ochraceus,

F. solani, F. moniliforme, F. culmorum.

21 showed the antifungal activity with a MIC value of 125

whereas other derivatives showed the antifungal activity against

F. culmorum with a MIC value of 250 g/mL.

mg/mL,

The most effective derivatives were also evaluated for their

cytotoxicity against C6 rat glioma cells. Compound 17 bearing 1-

phenyl-1H-tetrazole and nonsubstituted phenyl moieties was the

most promising anticancer agent against C6 glioma cell lines with

m

Compounds 2, 4, 5, 12, 13, 14, 24, 26, 27, 33, 34, 35, 36, 37, 38, 39,

40 and ketoconazole exhibited the inhibitory activity against Tri-

choderma harzianum with a MIC value of 250

rivatives exhibited the inhibitory activity with a MIC value of

125 g/mL.

The most effective derivatives were also evaluated for their

cytotoxicity against C6 rat glioma cells. The viability and IC₅₀values

m

g/mL, other de-

an IC₅₀value of 8.3 ꢀ 2.6

(IC₅₀¼ 13.7 ꢀ 1.2 g/mL). Compound 17 also exhibited low toxicity

g/mL. The re-

sults also revealed that compounds 16, 17 and 19 were the most

potent inhibitors of DNA synthesis against C6 cell lines.

mg/mL when compared with cisplatin

m

to NIH/3T3 cells with an IC₅₀value of 416.7 ꢀ 28.9

m

268

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

Table 4

The viability of compounds 1, 3, 8, 9, 10, 11, 16, 17, 19, 21, 22, 23, 24, 25, 27, 28, 29, 30, 35, 36, 37, 38, 39 against C6 cell lines.

Compounds

Concentration (

6

m

g/mL)^a

12

25

50

100

200

1

3

8

9

90.0 ꢀ 12.0

74.6 ꢀ 8.3

70.2 þ 1.6

85.2 ꢀ 3.4

80.4 ꢀ 4.6

88.0 ꢀ 9.3

63.5 þ 11.1

52.1 ꢀ 1.9

66.8 ꢀ 7.3

74.9 ꢀ 0.4

77.4 ꢀ 13.0

61.8 ꢀ 10.3

78.8 þ 17.1

72.4 ꢀ 2.9

65.9 ꢀ 6.4

57.9 ꢀ 8.4

74.3 ꢀ 18.1

82.8 ꢀ 10.0

93.2 ꢀ 20.7

89.6 þ 2.1

84.0 þ 16.7

95.9 þ 19.8

76.4 þ 26.0

57.3 þ 2.9

79.6 ꢀ 2.4

72.0 ꢀ 11.9

62.6 ꢀ 0.6

80.7 ꢀ 5.6

76.2 ꢀ 1.5

68.8 ꢀ 4.7

53.9 ꢀ 4.4

47.1 ꢀ 1.8

55.1 ꢀ 7.8

60.0 ꢀ 8.2

69.3 ꢀ 12.3

58.6 ꢀ 8.7

74.1 ꢀ 15.3

72.5 ꢀ 4.6

58.3 ꢀ 5.3

53.6 ꢀ 1.9

73.7 ꢀ 16.8

76.6 ꢀ 9.8

83.0 ꢀ 24.1

78.4 þ 8.1

77.4 þ 13.7

91.6 þ 11.9

72.1 þ 11.9

51.7 þ 2.8

77.3 ꢀ 3.6

70.7 ꢀ 6.5

58.8 ꢀ 2.3

64.2 ꢀ 7.9

69.3 ꢀ 8.1

60.7 ꢀ 6.5

43.9 ꢀ 6.2

42.5 ꢀ 6.3

38.8 ꢀ 3.1

49.3 ꢀ 6.5

62.3 ꢀ 12.4

47.9 ꢀ 4.1

64.2 ꢀ 5.9

65.9 ꢀ 4.8

50.1 ꢀ 3.7

42.6 ꢀ 9.1

50.1 ꢀ 16.5

53.3 ꢀ 15.1

82.7 ꢀ 10.7

74.0 þ 8.4

63.9 þ 3.3

91.1 þ 24.6

72.3 þ 23.6

31.2 þ 8.8

62.4 ꢀ 13.4

53.8 ꢀ 18.9

58.8 ꢀ 3.9

56.2 ꢀ 1.0

69.2 ꢀ 1.8

44.3 ꢀ 3.7

35.6 ꢀ 4.6

36.7 ꢀ 9.1

29.5 ꢀ 3.4

43.7 ꢀ 4.5

60.0 ꢀ 6.8

32.0 ꢀ 4.9

61.1 ꢀ 6.3

63.9 ꢀ 1.7

40.6 ꢀ 5.2

31.5 ꢀ 3.3

37.4 ꢀ 7.1

37.9 ꢀ 3.4

64.4 ꢀ 2.3

66.5 þ 10.7

55.5 þ 1.4

61.8 þ 2.4

69.0 þ 15.7

27.2 þ 4.1

38.6 ꢀ 8.8

39.6 ꢀ 7.6

55.2 ꢀ 5.4

38.8 ꢀ 8.3

64.1 ꢀ 4.9

39.7 ꢀ 5.7

30.9 ꢀ 3.6

35.8 ꢀ 5.9

28.5 ꢀ 4.6

35.8 ꢀ 3.5

54.3 ꢀ 3.4

24.3 ꢀ 2.4

59.8 ꢀ 5.9

61.8 ꢀ 4.8

31.5 ꢀ 11.1

25.9 ꢀ 5.1

34.9 ꢀ 11.1

34.1 ꢀ 5.2

45.9 ꢀ 4.4

51.5 þ 8.1

53.4 þ 0.4

45.5 þ 6.6

67.6 þ 14.9

17.3 þ 2.5

35.5 ꢀ 3.7

27.8 ꢀ 3.4

52.6 ꢀ 2.7

26.8 ꢀ 2.2

58.6 ꢀ 5.0

39.7 ꢀ 6.5

28.6 ꢀ 4.3

22.6 ꢀ 0.9

22.2 ꢀ 3.9

30.6 ꢀ 4.3

52.4 ꢀ 2.6

20.4 ꢀ 2.1

54.0 ꢀ 2.4

59.2 ꢀ 1.9

13.3 ꢀ 2.8

22.2 ꢀ 6.2

26.5 ꢀ 6.3

29.0 ꢀ 5.1

43.7 þ 9.7

37.6 þ 6.6

32.2 þ 6.2

39.7 þ 9.4

63.2 þ 15.1

16.2 þ 2.4

10

11

16

17

19

21

22

23

24

25

27

28

29

30

35

36

37

38

39

Cisplatin

a

Viability % was determined relative to controls.

In the view of the current study, further research can be carried

out on the development of new effective antimicrobial and anti-

cancer agents by the modiﬁcation of these compounds.

5.1.1.3.1. 1-[(1-Methyl-1H-tetrazol-5-yl)thioacetyl]-4-phenyl thi-

osemicarbazide (C-2). M.p.: 168e169 ^ꢁC.

IR (KBr) n_max(cm^ꢂ1): 3296.12, 3236.33, 3215.11 (NeH stretch-

ing), 3056.96 (Aromatic CeH stretching), 2975.96, 2937.38

(Aliphatic CeH stretching), 1685.67 (C]O stretching), 1593.09,

1548.73, 1537.16, 1502.44, 1471.59 (C]N, C]C stretching and NeH

bending), 1446.51 (CeH bending), 1269.07 (CeN stretching),

1207.36, 1153.35 (Aromatic CeH in plane bending), 752.19, 703.97

(Aromatic CeH out of plane bending).

5. Experimental

5.1. Chemistry

All reagents were purchased from commercial suppliers and

were used without further puriﬁcation. Melting points were

determined on an Electrothermal 9100 melting point apparatus

(Weiss-Gallenkamp, Loughborough, UK) and are uncorrected. IR

spectra were recorded on Shimadzu 8400 FT-IR spectrophotometer

(Shimadzu, Tokyo, Japan). ¹H NMR spectra were recorded on a

Bruker 300 MHz and 400 MHz spectrometer (Bruker, Billerica, USA),

respectively. ¹³C NMR spectra were recorded on a Bruker 100 MHz

spectrometer (Bruker, Billerica, USA). Mass spectra were recorded

on an Agilent LC-MSD-Trap-SL Mass spectrometer (Agilent, Min-

nesota, USA). Elemental analyses were performed on a Perkine

Elmer EAL 240 elemental analyser (PerkineElmer, Norwalk, USA).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 3.93 (3H, s, CH₃), 4.11

(2H, s, SeCH₂), 7.13 (1H, t, J₁¼ 7.6 Hz, J₂¼ 6.8 Hz, phenyl H₄),

7.30 (2H, t, J₁¼ 8 Hz, J₂¼ 7.6 Hz, phenyl H₃, H₅), 7.43 (2H, d,

J ¼ 7.6 Hz, phenyl H₂, H₆), 9.59 (1H, s, NeH), 9.72 (1H, s, NeH), 10.40

(1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.48 (CH₃), 36.00

(CH₂), 126.01 (2CH), 128.82 (3CH), 139.50 (C), 154.02 (C), 167.11 (C),

181.68 (C).

MS (ESI)(m/z): (M^þꢂ 1) 322.

For C₁₁H₁₃N₇OS₂calculated: C, 40.85; H, 4.05; N, 30.32; Found:

C, 40.84; H, 4.06; N, 30.32.

5.1.1.3.2. 1-[(Pyrimidin-2-yl)thioacetyl]-4-phenyl

semicarbazide (C-5). M.p.: 176e181 ^ꢁC.

thio-

5.1.1. General procedure for the synthesis of the compounds

5.1.1.1. Ethyl 2-[(aryl)thio]acetate derivatives (A1e6). A mixture of

thiol (0.2 mol) and ethyl chloroacetate (0.2 mol) in the presence of

potassium carbonate (0.2 mol) in acetone (60 mL) was reﬂuxed for

24 h. The reaction mixture was cooled, ﬁltered and the crude

product was solved in water and then extracted with diethyl ether

[31e34].

IR (KBr) n_max(cm^ꢂ1): 3307.69, 3261.40 (NeH stretching),

3103.25 (Aromatic CeH stretching), 2933.53 (Aliphatic CeH

stretching), 1681.81 (C]O stretching), 1596.95, 1508.23, 1450.37

(C]N, C]C stretching and NeH bending), 1379.01 (CeH bending),

1317.29, 1274.86 (CeN stretching), 1213.14, 1159.14 (Aromatic CeH

in plane bending), 636.47 (CeS stretching).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 3.99 (2H, s, SeCH₂), 7.14

5.1.1.2. 2-[(Aryl)thio]acetohydrazide derivatives (B1e6). A mixture

of the ester (A) (0.1 mol) and hydrazine hydrate (0.2 mol) in ethanol

(40 mL) was stirred at room temperature for 10 h and then ﬁltered

[31e34].

(1H, t, J₁¼ 7.6 Hz, J₂¼ 6.8 Hz, phenyl H₄), 7.18 (1H, t, J₁, J₂¼ 4.8 Hz,

pyrimidine H₄), 7.31 (2H, t, J₁¼ 8.4 Hz, J₂¼ 7.2 Hz, phenyl H₃, H₅),

7.39 ve 7.48 (2H, 2d, J₁, J₂¼ 7.6 Hz, phenyl H₂, H₆), 8.58 (2H, d,

J ¼ 4.8 Hz, pyrimidine), 9.55 (1H, s, NeH), 9.70 (1H, s, NeH), 10.26

(1H, s, NeH).

5.1.1.3. 1-(Arylthioacetyl)-4-phenyl thiosemicarbazide derivatives

(C1e6). A mixture of the hydrazide (B) (0.05 mol) and phenyl

isothiocyanate (0.05 mol) in ethanol (30 mL) was reﬂuxed for 1 h

and then ﬁltered [34e36].

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 33.64 (CH₂), 118.10

(CH), 125.92 (2CH), 126.53 (CH), 128.81 (2CH), 139.66 (C), 158.48

(2CH), 168.13 (C), 170.87 (C), 181.52 (C).

MS (ESI)(m/z): (M^þꢂ 1) 318.

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

269

Table 5

(2

mmol)

and

2-bromoacetophenone/4⁰-substituted-2-

IC₅₀values of the compounds against NIH/3T3 and C6 cell lines.

bromoacetophenone (2 mmol) in ethanol (15 mL) was reﬂuxed

for 5 h and then ﬁltered [18,36].

Compound

IC₅₀value (mg/mL)

5.1.1.4.1. N⁰-[(3,4-Diphenylthiazol-2(3H)-ylidene)]-2-[(4-methyl-

4H-1,2,4-triazol-3-yl)thio]acetohydrazide (1). IR (KBr) n_max(cm^ꢂ1):

2927.82 (Aliphatic CeH stretching), 1677.23 (C]O stretching),

1572.17, 1511.84, 1488.62 (C]N, C]C stretching and NeH bending),

1442.44, 1356.41 (CeH bending), 1299.82 (CeN stretching),

1198.50, 1155.96, 1092.80, 1072.12, 1011.25 (Aromatic CeH in plane

bending), 917.04, 830.77, 760.55, 691.98 (Aromatic CeH out of plane

bending and CeS stretching).

NIH/3T3 cell lines

C6 cell lines

1

2

3

4

5

6

7

8

216.7 ꢀ 15.3

133.3 ꢀ 11.5

130.0 ꢀ 20.0

166.7 ꢀ 5.8

65.0 ꢀ 17.3

51.7 ꢀ 20.8

58.3 ꢀ 36.8

153.3 ꢀ 5.7

173.3 ꢀ 20.8

166.7 ꢀ 28.8

243.3 ꢀ 40.4

45.0 ꢀ 5.0

113.3 ꢀ 52.0

40.0 ꢀ 13.2

19.3 ꢀ 0.6

213.3 ꢀ 5.7

416.7 ꢀ 28.9

14.3 ꢀ 4.0

>500

11.7 ꢀ 1.5

206.7 ꢀ 15.3

313.3 ꢀ 15.3

433.3 ꢀ 28.9

453.3 ꢀ 53.3

196.7 ꢀ 5.8

176.7 ꢀ 86.6

373.3 ꢀ 30.5

300.0 ꢀ 50.0

383.3 ꢀ 28.9

253.3 ꢀ 25.2

55.0 ꢀ 5.0

<3.9

68.3 ꢀ 13.1

nt

56.0 ꢀ 10.4

nt

>200

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 3.60 (3H, s, CH₃), 3.86

9

67.7 ꢀ 8.8

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

Cisplatin

>200

(2H, s, SeCH₂), 6.54 (1H, s, thiazoline), 7.13e7.42 (10H, m, phenyl),

8.59 (1H, s, triazole), 10.36 (1H, s, NeH).

36.3 ꢀ 4.9

nt

23.3 ꢀ 6.2

8.3 ꢀ 2.6

nt

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 31.61 (CH₃), 36.28

(CH₂), 99.85 (CH), 128.46 (CH), 128.85 (2CH), 128.94 (CH), 129.17

(CH), 129.23 (CH), 129.29 (2CH), 129.54 (2CH), 131.31 (C), 138.02 (C),

140.21 (CH), 146.90 (C), 149.43 (2C), 167.90 (C).

MS (ESI)(m/z): (M^þꢂ 1) 421.

For C₂₀H₁₈N₆OS₂calculated: C, 56.85; H, 4.29; N,19.89; Found: C,

56.85; H, 4.30; N, 19.90.

14.7 ꢀ 3.4

nt

5.1.1.4.2. N⁰-[(4-Nitrophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(4-methyl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (2). IR (KBr)

n_max(cm^ꢂ1): 3105.32 (Aromatic CeH stretching), 1688.90 (C]O

stretching), 1601.78, 1587.64, 1574.24, 1556.46, 1488.71, 1454.19

(C]N, C]C stretching and NeH bending), 1445.83, 1341.62 (CeH

bending), 1318.20, 1300.36, 1287.44 (CeN stretching), 1195.46,

1173.07, 1158.41, 1107.02, 1075.43, 1028.28, 1002.02 (Aromatic CeH

in plane bending), 921.02, 852.97, 765.46, 753.34, 691.94 (Aromatic

CeH out of plane bending and CeS stretching).

24.3 ꢀ 6.3

>200

21.0 ꢀ 3.7

>200

nt

25.3 ꢀ 6.1

18.7 ꢀ 2.9

27.3 ꢀ 10.5

28.0 ꢀ 8.6

nt

85.0 ꢀ 10.8

101.7 ꢀ 16.5

110.0 ꢀ 8.2

83.3 ꢀ 12.5

>200

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.46 and 3.56 (3H, 2s,

106.7 ꢀ 30.5

193.3 ꢀ 11.5

>500

CH₃), 4.06 (2H, s, SeCH₂), 6.70e8.54 (11H, m, thiazoline, phenyl, 4-

nitrophenyl, triazole), 11.30 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 31.40 (CH₃), 35.32

450 ꢀ 50

(CH₂), 107.11 (CH), 121.51 (CH), 124.13 (CH), 124.19 (CH), 124.44

(2CH),128.97 (CH),129.07 (CH),129.68 (CH),129.72 (CH),137.59 (C),

138.56 (C), 143.37 (CH), 146.84 (C), 147.57 (C), 147.90 (C), 150.68 (C),

168.04 (C).

>500

396.7 ꢀ 5.8

273.3 ꢀ 20.8

<3.9

nt

13.7 ꢀ 1.2

MS (ESI)(m/z): (M^þꢂ 1) 466.

nt: not tested.

For C₂₀H₁₇N₇O₃S₂calculated: C, 51.38; H, 3.67; N, 20.97; Found:

C, 51.40; H, 3.68; N, 20.97.

5.1.1.4.3. N⁰-[(4-Methylphenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(4-methyl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (3). IR (KBr)

n_max(cm^ꢂ1): 2920.48 (Aliphatic CeH stretching), 1676.49 (C]O

stretching), 1598.24, 1557.20, 1497.37, 1451.91 (C]N, C]C stretch-

ing and NeH bending), 1354.97 (CeH bending), 1286.79 (CeN

stretching), 1183.91, 1111.26, 1065.51, 1017.97 (Aromatic CeH in

plane bending), 967.30, 812.24, 783.89, 754.20, 693.09 (Aromatic

CeH out of plane bending and CeS stretching).

For C₁₃H₁₃N₅OS₂calculated: C, 48.88; H, 4.10; N, 21.93; Found: C,

48.85; H, 4.10; N, 21.95.

5.1.1.4. N⁰-(3,4-Diarylthiazol-2(3H)-ylidene)-2-(arylthio)acetohy-

drazide derivatives (1e40). A mixture of the thiosemicarbazide (C)

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 2.31 (3H, s, CH₃), 3.53

(3H, s, CH₃, triazole), 4.02 (2H, s, SeCH₂), 7.04 (1H, s, thiazoline),

7.23 (2H, d, J ¼ 8 Hz, 4-methylphenyl), 7.35e7.53 (7H, m, phenyl, 4-

methylphenyl), 8.85 (1H, s, triazole), 10.35 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 21.61 (CH₃), 32.07

(CH₃), 35.48 (CH₂), 101.82 (CH), 124.17 (CH), 124.87 (2CH), 128.44

(C), 129.01 (2CH), 130.01 (2CH), 130.95 (2CH), 139.73 (C), 140.69 (C),

141.10 (CH), 146.83 (C), 149.79 (2C), 167.68 (C).

MS (ESI)(m/z): (M^þþ 1) 437.

For C₂₁H₂₀N₆OS₂calculated: C, 57.78; H, 4.62; N,19.25; Found: C,

57.79; H, 4.60; N, 19.25.

5.1.1.4.4. N⁰-[(4-Methoxyphenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(4-methyl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (4). IR (KBr)

n_max(cm^ꢂ1): 3355.05 (NeH stretching), 3054.18 (Aromatic CeH

stretching), 2871.63 (Aliphatic CeH stretching), 1705.44 (C]O

stretching), 1603.12, 1561.21, 1510.66, 1492.71, 1466.63 (C]N, C]C

Fig. 1. DNA synthesis inhibitory activity of compounds 16, 17, 19, 21, 23, 27, 28, 29, 30

and cisplatin on C6 cells.

270

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

stretching and NeH bending), 1438.95, 1416.76, 1346.04 (CeH

bending), 1300.05, 1258.52, 1230.96 (CeN stretching), 1197.99 (CeO

stretching), 1181.13, 1107.16, 1077.25, 1051.47, 1036.95, 1023.50

(Aromatic CeH in plane bending), 961.96, 878.09, 846.37, 832.55,

819.84, 774.79, 766.03, 744.95, 714.93, 694.16, 656.22 (Aromatic Ce

H out of plane bending and CeS stretching).

and NeH bending), 1422.99, 1393.45, 1351.23, 1374.05 (CeH

bending), 1310.04, 1225.98 (CeN stretching), 1190.48, 1165.12,

1109.10, 1070.32, 1050.98, 1009.23 (Aromatic CeH in plane

bending), 963.01, 945.29, 895.41, 866.11, 827.64, 818.18, 782.74,

760.36, 724.31, 714.07, 693.63, 682.24 (Aromatic CeH out of plane

bending and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 3.57 (3H, s, CH₃), 3.82

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.72 and 3.73 (3H, 2s,

(3H, s, OCH₃), 4.01 (2H, s, SeCH₂), 6.95 (1H, s, thiazoline), 7.02 (2H,

d, J ¼ 9 Hz, 4-methoxyphenyl), 7.38e7.57 (7H, m, phenyl, 4-

methoxyphenyl), 8.75 (1H, s, triazole), 12.08 (1H, s, NeH).

CH₃), 4.01 and 4.10 (2H, 2s, SeCH₂), 7.14e7.51 (10H, m, thiazoline,

phenyl, 4-bromophenyl), 9.29 and 9.38 (1H, 2s, triazole), 11.14 (1H,

s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d

(ppm): 31.95 (CH₃), 35.35

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 32.76 (CH₃), 35.42

(CH₂), 56.07 (CH₃), 100.77 (CH), 114.91 (2CH), 119.98 (CH), 124.11

(C), 128.32 (2CH), 130.72 (2CH), 130.96 (2CH), 140.06 (C), 140.94

(CH), 146.90 (C), 149.70 (2C), 161.25 (C), 167.75 (C).

(CH₂), 105.53 (CH), 123.66 (C), 128.85 (CH), 129.03 (2CH), 129.40

(2CH), 129.77 (C), 130.61 (2CH), 131.85 (CH), 131.99 (CH), 135.19 (C),

140.95 (CH), 146.39 (C), 151.29 (2C), 165.81 (C).

MS (ESI)(m/z): (M^þþ 1) 453.

MS (ESI)(m/z): (M^þþ 1) 501.

For C₂₁H₂₀N₆O₂S₂calculated: C, 55.73; H, 4.45; N, 18.57; Found:

C, 55.75; H, 4.45; N, 18.58.

For C₂₀H₁₇BrN₆OS₂calculated: C, 47.91; H, 3.42; N, 16.76; Found:

C, 47.90; H, 3.40; N, 16.75.

5.1.1.4.5. N⁰-[(4-Fluorophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(4-methyl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (5). IR (KBr)

n_max(cm^ꢂ1): 3421.07 (NeH stretching), 3061.60 (Aromatic CeH

stretching), 2890.79 (Aliphatic CeH stretching), 1704.58 (C]O

stretching), 1600.44, 1584.15, 1567.31, 1505.80, 1496.56, 1467.04

(C]N, C]C stretching and NeH bending), 1410.18, 1380.91,

1344.37 (CeH bending), 1311.88, 1276.96, 1232.00 (CeN stretch-

ing), 1200.92, 1162.99, 1103.17, 1028.52, 1013.09 (Aromatic CeH in

plane bending), 962.40, 832.02, 780.42, 763.32, 747.72, 712.36,

689.84, 654.90 (Aromatic CeH out of plane bending and CeS

stretching).

5.1.1.4.8. N⁰-[(4-Cyanophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(4-methyl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (8). IR (KBr)

n_max(cm^ꢂ1): 3433.06 (NeH stretching), 3037.68 (Aromatic CeH

stretching), 2908.45 (Aliphatic CeH stretching), 2229.56 (C^N

stretching), 1716.53 (C]O stretching), 1598.88,1488.94 (C]N, C]C

stretching and NeH bending), 1317.29 (CeN stretching), 1110.92

(Aromatic CeH in plane bending), 833.19 (Aromatic CeH out of

plane bending).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 3.76 (3H, s, CH₃), 4.06

and 4.15 (2H, 2s, SeCH₂), 7.28e7.52 (8H, m, thiazoline, phenyl, 4-

cyanophenyl), 7.76e7.81 (2H, m, 4-cyanophenyl), 9.52 and 9.56

(1H, 2s, triazole), 11.15 (1H, s, NeH).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.59 and 3.68 (3H, 2s,

CH₃), 4.03 (2H, d, J ¼ 6 Hz, SeCH₂), 7.05e7.57 (10H, m, thiazoline,

phenyl, 4-ﬂuorophenyl), 8.84 and 9.02 (1H, 2s, triazole), 12.10 (1H,

s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 32.29 (CH₃), 34.72

(CH₂), 106.14 (CH), 111.64 (C), 118.01 (C), 128.19 (CH), 128.54 (2CH),

129.01 (2CH), 129.73 (CH), 129.79 (CH), 131.05 (2CH), 132.08 (C),

136.55 (C), 139.40 (CH), 145.35 (C), 145.44 (C), 151.05 (C), 164.69 (C).

MS (ESI)(m/z): (M^þþ 1) 448.

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 32.28 (CH₃), 35.47

(CH₂), 102.79 (CH), 116.49 (CH), 116.71 (CH), 124.11 (CH), 124.14

(CH), 124.23 (CH), 128.59 (C), 131.02 (2CH), 131.80 (CH), 131.89 (CH),

139.54 (C), 140.02 (CH), 146.74 (C), 150.14 (C), 150.99 (C), 162.50 (C),

167.65 (C).

For C₂₁H₁₇N₇OS₂calculated: C, 56.36; H, 3.83; N, 21.91; Found: C,

56.37; H, 3.83; N, 21.90.

5.1.1.4.9. N⁰-[(3,4-Diphenylthiazol-2(3H)-ylidene)]-2-[(1-methyl-

1H-tetrazol-5-yl)thio]acetohydrazide (9). IR (KBr) n_max(cm^ꢂ1):

3053.11 (Aromatic CeH stretching), 2860.16 (Aliphatic CeH

stretching), 1723.44 (C]O stretching), 1595.18, 1573.85, 1557.58,

1514.48, 1458.36 (C]N, C]C stretching and NeH bending),

1445.50, 1397.08, 1368.05 (CeH bending), 1310.69, 1227.67 (CeN

stretching), 1210.93, 1194.17, 1179.66, 1124.94, 1076.71, 1019.72,

1003.86 (Aromatic CeH in plane bending), 963.44, 936.93, 917.06,

896.68, 835.43, 811.36, 776.09, 745.91, 691.14 (Aromatic CeH out of

plane bending and CeS stretching).

MS (ESI)(m/z): (M^þþ 1) 441.

For C₂₀H₁₇FN₆OS₂calculated: C, 54.53; H, 3.89; N, 19.08; Found:

C, 54.54; H, 3.90; N, 19.08.

5.1.1.4.6. N⁰-[(4-Chlorophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(4-methyl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (6). IR (KBr)

n_max(cm^ꢂ1): 3037.87 (Aromatic CeH stretching), 2851.33 (Aliphatic

CeH stretching), 1718.10 (C]O stretching), 1585.11, 1553.42,

1505.64, 1487.76, 1453.15 (C]N, C]C stretching and NeH

bending), 1422.87, 1395.31, 1376.38 (CeH bending), 1310.58 (CeN

stretching), 1190.98, 1164.76, 1110.75, 1090.99, 1051.16, 1013.89

(Aromatic CeH in plane bending), 945.69, 963.72, 895.74, 866.25,

829.02, 820.88, 783.59, 760.19, 710.09, 696.14, 684.73 (Aromatic Ce

H out of plane bending and CeS stretching).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 3.84 and 3.95 (3H, 2s,

CH₃), 4.20 and 4.23 (2H, 2s, SeCH₂), 7.17 (1H, s, thiazoline), 7.39e

7.57 (10H, m, phenyl), 11.38 and 12.41 (1H, 2s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.44 (CH₃), 35.75

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 3.71 and 3.72 (3H, 2s,

(CH₂), 103.23 (CH), 124.57 (CH), 127.38 (2CH), 128.97 (CH), 129.06

(CH), 129.17 (CH), 129.26 (2CH), 129.41 (2CH), 130.99 (C), 131.10 (C),

141.21 (2C), 153.37 (C), 167.12 (C).

CH₃), 3.87 and 4.00 (2H, 2s, SeCH₂), 7.17e7.41 (10H, m, thiazoline,

phenyl, 4-chlorophenyl), 9.04 (1H, s, triazole), 11.80 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d

(ppm): 32.95 (CH₃), 35.39

MS (ESI)(m/z): (M^þþ 1) 424.

(CH₂), 106.05 (CH), 128.37 (2CH), 129.03 (CH), 129.09 (2CH), 130.69

(C), 130.91 (2CH), 131.77 (2CH), 134.99 (2C), 141.07 (CH), 146.33 (C),

151.57 (2C), 166.05 (C).

For C₁₉H₁₇N₇OS₂calculated: C, 53.88; H, 4.05; N, 23.15; Found: C,

53.90; H, 4.03; N, 23.15.

5.1.1.4.10. N⁰-[(4-Nitrophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(1-methyl-1H-tetrazol-5-yl)thio]acetohydrazide (10). IR (KBr) n_max

(cm^ꢂ1): 3197.61 (NeH stretching), 1721.69 (C]O stretching),

1589.15, 1552.80, 1516.69, 1489.05, 1455.35 (C]N, C]C stretching

and NeH bending), 1408.01, 1387.21, 1345.52 (CeH bending),

1288.86 (CeN stretching), 1172.14, 1108.09, 1075.04, 1014.94 (Aro-

matic CeH in plane bending), 967.57, 915.36, 853.61, 758.06,

738.42, 697.12 (Aromatic CeH out of plane bending and CeS

stretching).

MS (ESI)(m/z): (M^þþ 1) 457.

For C₂₀H₁₇ClN₆OS₂calculated: C, 52.57; H, 3.75; N, 18.39; Found:

C, 52.58; H, 3.75; N, 18.40.

5.1.1.4.7. N⁰-[(4-Bromophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(4-methyl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (7). IR (KBr)

n_max(cm^ꢂ1): 3108.56, 3035.01 (Aromatic CeH stretching), 2850.52,

2734.33 (Aliphatic CeH stretching), 1717.73 (C]O stretching),

1580.57, 1553.70, 1505.47, 1485.83, 1452.55 (C]N, C]C stretching

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

271

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 3.87 (3H, s, CH₃), 4.08

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 3.91 (3H, s, CH₃), 4.19

(2H, s, SeCH₂), 6.76e8.25 (10H, m, thiazoline, phenyl, 4-

nitrophenyl), 11.81 (1H, s, NeH).

and 4.24 (2H, 2s, SeCH₂), 6.99 (1H, s, thiazoline), 7.25e7.57 (9H, m,

phenyl, 4-ﬂuorophenyl), 12.04 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d

(ppm): 34.34 (CH₃), 35.69

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.40 (CH₃), 35.60

(CH₂), 104.06 (CH), 121.84 (CH), 122.21 (CH), 123.66 (CH), 124.13

(CH), 124.38 (CH), 128.84 (CH), 128.93 (CH), 129.13 (CH), 129.25

(CH), 137.55 (C), 138.34 (C), 147.57 (C), 148.03 (C), 148.16 (C), 153.24

(C), 167.37 (C).

(CH₂), 102.50 (CH), 116.63 (2CH), 124.01 (CH), 124.40 (2CH), 128.81

(C), 131.09 (2CH), 131.79 (CH), 131.89 (CH), 140.19 (3C), 153.37 (C),

162.52 (C), 167.17 (C).

MS (ESI)(m/z): (M^þþ 1) 442.

MS (ESI)(m/z): (M^þþ 1) 469.

For C₁₉H₁₆FN₇OS₂calculated: C, 51.69; H, 3.65; N, 22.21; Found:

C, 51.70; H, 3.65; N, 22.20.

For C₁₉H₁₆N₈O₃S₂calculated: C, 48.71; H, 3.44; N, 23.92; Found:

C, 48.70; H, 3.45; N, 23.90.

5.1.1.4.14. N⁰-[(4-Chlorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-methyl-1H-tetrazol-5-yl)thio]acetohydrazide (14). IR (KBr)

n_max(cm^ꢂ1): 2879.95 (Aliphatic CeH stretching), 1715.30 (C]O

stretching), 1571.56, 1458.25 (C]N, C]C stretching and NeH

bending), 1092.79, 1011.35 (Aromatic CeH in plane bending),

752.01 (Aromatic CeH out of plane bending).

5.1.1.4.11. N⁰-[(4-Methylphenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-methyl-1H-tetrazol-5-yl)thio]acetohydrazide (11). IR (KBr)

n_max(cm^ꢂ1): 3033.02 (Aromatic CeH stretching), 2861.21 (Aliphatic

CeH stretching), 1715.69 (C]O stretching), 1604.28, 1570.13,

1509.78, 1497.50, 1457.38 (C]N, C]C stretching and NeH

bending), 1387.32, 1359.70 (CeH bending), 1298.45, 1281.47,

1261.97, 1246.99, 1230.14 (CeN stretching), 1192.95, 1171.02,

1127.83,1077.32,1018.13,1005.46 (Aromatic CeH in plane bending),

959.96, 883.71, 842.61, 827.41, 812.77, 794.83, 782.15, 772.95,

753.42, 736.62, 698.75 (Aromatic CeH out of plane bending and Ce

S stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.91 (3H, s, CH₃), 4.23

(2H, d, J ¼ 12 Hz, SeCH₂), 7.03 (1H, s, thiazoline), 7.33e7.56 (9H, m,

phenyl, 4-chlorophenyl), 12.19 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.39 (CH₃), 35.49

(CH₂), 102.40 (CH), 124.11 (2CH), 126.63 (CH), 128.34 (2CH), 129.51

(C), 130.94 (2CH), 131.03 (2CH), 135.72 (2C), 139.82 (2C), 153.41 (C),

167.25 (C).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 2.36 (3H, s, CH₃), 3.89

(3H, s, CH₃, tetrazole), 4.24 and 4.32 (2H, 2s, SeCH₂), 7.10 (1H, s,

thiazoline), 7.25 (2H, d, J ¼ 8.4 Hz, aromatic), 7.37 (2H, d, J ¼ 7.6 Hz,

aromatic), 7.43e7.48 (3H, m, aromatic), 7.57e7.61 (2H, m, aro-

matic), 12.34 (1H, s, NeH).

MS (ESI)(m/z): (M^þþ 1) 458.

For C₁₉H₁₆ClN₇OS₂calculated: C, 49.83; H, 3.52; N, 21.41; Found:

C, 49.82; H, 3.50; N, 21.42.

5.1.1.4.15. N⁰-[(4-Bromophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-methyl-1H-tetrazol-5-yl)thio]acetohydrazide (15). IR (KBr)

n_max(cm^ꢂ1): 3049.57 (Aromatic CeH stretching), 2878.02

(Aliphatic CeH stretching), 1710.14 (C]O stretching), 1609.00,

1600.29, 1571.18, 1488.78, 1463.73 (C]N, C]C stretching and NeH

bending), 1398.02 (CeH bending), 1286.25, 1230.88 (CeN stretch-

ing), 1195.72, 1170.19, 1115.87, 1071.96, 1027.90, 1004.77 (Aromatic

CeH in plane bending), 958.17, 915.24, 887.94, 827.97, 805.46,

748.09, 713.05, 700.50, 690.56 (Aromatic CeH out of plane bending

and CeS stretching).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 21.64 (CH₃), 34.39

(CH₃), 35.70 (CH₂), 102.11 (CH), 124.46 (CH), 124.69 (2CH), 128.85

(C), 129.07 (2CH), 129.97 (2CH), 131.09 (2CH), 138.96 (C), 140.76

(2C), 141.30 (C), 153.35 (C), 167.12 (C).

MS (ESI)(m/z): (M^þþ 1) 438.

For C₂₀H₁₉N₇OS₂calculated: C, 54.90; H, 4.38; N, 22.41; Found:

C, 54.89; H, 4.35; N, 22.42.

5.1.1.4.12. N⁰-[(4-Methoxyphenyl-3-phenylthiazol-2(3H)-yli-

dene)]-2-[(1-methyl-1H-tetrazol-5-yl)thio]acetohydrazide

(12).

IR (KBr) n_max(cm^ꢂ1): 3360.96 (NeH stretching), 2713.08 (Aliphatic

CeH stretching), 1699.11 (C]O stretching), 1601.61, 1573.64,

1557.76,1508.14,1456.13 (C]N, C]C stretching and NeH bending),

1410.34 (CeH bending), 1299.85, 1256.06 (CeN stretching), 1204.33

(CeO stretching), 1179.66, 1135.14, 1115.76, 1023.98 (Aromatic CeH

in plane bending), 962.51, 839.78, 816.69, 766.97, 720.80, 697.73

(Aromatic CeH out of plane bending and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.91 (3H, s, CH₃), 4.24

(2H, d, J ¼ 9 Hz, SeCH₂), 7.03 (1H, s, thiazoline), 7.33e7.66 (9H, m,

phenyl, 4-bromophenyl), 12.05 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.41 (CH₃), 35.56

(CH₂), 103.04 (CH), 124.30 (C), 124.59 (CH), 126.79 (2CH), 128.65

(2CH),131.06 (C), 131.16 (2CH),132.43 (2CH),139.96 (3C),153.40 (C),

167.22 (C).

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 3.69 and 3.81 (3H, 2s,

MS (ESI)(m/z): (M^þþ 1) 503.

OCH₃), 3.90 and 3.98 (3H, 2s, CH₃), 4.12e4.22 (2H, m, SeCH₂), 6.65e

7.60 (10H, m, thiazoline, phenyl, 4-methoxyphenyl), 11.81 (1H, s,

NeH).

For C₁₉H₁₆BrN₇OS₂calculated: C, 45.42; H, 3.21; N,19.52; Found:

C, 45.42; H, 3.21; N, 19.52.

5.1.1.4.16. N⁰-[(4-Cyanophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-methyl-1H-tetrazol-5-yl)thio]acetohydrazide (16). IR (KBr)

n_max(cm^ꢂ1): 3429.20 (NeH stretching), 3058.89 (Aromatic CeH

stretching), 2894.95 (Aliphatic CeH stretching), 2227.63 (C^N

stretching), 1720.39 (C]O stretching), 1604.66, 1571.88, 1477.37

(C]N, C]C stretching and NeH bending), 1176.50 (Aromatic CeH

in plane bending), 856.34 (Aromatic CeH out of plane bending),

667.32 (CeS stretching).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.34 (CH₃), 35.58

(CH₂), 55.96 (CH₃), 99.90 (CH), 114.74 (2CH),123.36 (CH),129.20 (C),

130.09 (2CH), 130.35 (2CH), 130.70 (2CH), 130.86 (C), 140.50 (2C),

153.38 (C), 160.96 (C), 167.17 (C).

MS (ESI)(m/z): (M^þþ 1) 454.

For C₂₀H₁₉N₇O₂S₂calculated: C, 52.96; H, 4.22; N, 21.62; Found:

C, 52.95; H, 4.20; N, 21.63.

5.1.1.4.13. N⁰-[(4-Fluorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-methyl-1H-tetrazol-5-yl)thio]acetohydrazide (13). IR (KBr)

n_max(cm^ꢂ1): 3503.26 (NeH stretching), 3116.58 (Aromatic CeH

stretching), 1715.79 (C]O stretching), 1602.24, 1566.64, 1496.42,

1455.49 (C]N, C]C stretching and NeH bending),1407.64,1371.82

(CeH bending), 1278.53,1228.28 (CeN stretching), 1173.37, 1160.55,

1101.28,1075.68,1025.17,1010.42 (Aromatic CeH in plane bending),

963.54, 889.11, 855.21, 822.06, 792.38, 777.12, 757.78, 745.09,

703.94, 684.82, 656.09 (Aromatic CeH out of plane bending and Ce

S stretching).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 3.91 (3H, s, CH₃), 4.22

(2H, d, J ¼ 16 Hz, SeCH₂), 6.96e7.11 (1H, thiazoline), 7.28e8.04 (9H,

m, phenyl, 4-cyanophenyl), 12.02 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 33.56 (CH₃), 34.56

(CH₂), 110.88 (CH), 112.20 (C), 118.25 (C), 122.67 (CH), 128.77 (2CH),

128.93 (2CH), 130.08 (2CH), 131.44 (CH), 131.80 (CH), 132.44 (C),

138.22 (3C), 152.55 (C), 166.47 (C).

MS (ESI)(m/z): (M^þþ 1) 449.

For C₂₀H₁₆N₈OS₂calculated: C, 53.56; H, 3.60; N, 24.98; Found:

C, 53.55; H, 3.63; N, 24.98.

272

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

5.1.1.4.17. N⁰-[(3,4-Diphenylthiazol-2(3H)-ylidene)]-2-[(1-phenyl-

stretching), 1599.73, 1575.50, 1561.78, 1497.12, 1457.11 (C]N, C]C

stretching and NeH bending), 1442.55, 1427.94, 1403.56, 1385.67

(CeH bending),1317.02,1280.76,1249.12,1239.06 (CeN stretching),

1203.67 (CeO stretching), 1181.70, 1155.57, 1110.39, 1079.63,

1063.49, 1041.70, 1024.21 (Aromatic CeH in plane bending), 986.34,

964.04, 916.01, 889.10, 836.98, 812.86, 782.06, 761.19, 744.75,

716.73, 699.26, 685.00 (Aromatic CeH out of plane bending and Ce

S stretching).

1H-tetrazol-5-yl)thio]acetohydrazide (17). IR (KBr) n_max(cm^ꢂ1):

3108.98 (Aromatic CeH stretching), 1671.06 (C]O stretching),

1616.69, 1578.50, 1551.16, 1497.07 (C]N, C]C stretching and NeH

bending), 1445.30, 1386.11, 1357.86 (CeH bending), 1300.61,

1242.75 (CeN stretching), 1155.47, 1090.59, 1074.06, 1014.15 (Aro-

matic CeH in plane bending), 966.93, 916.68, 830.89, 758.47, 691.94

(Aromatic CeH out of plane bending and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 4.20e4.29 (2H, m, Se

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.78 (3H, s, OCH₃), 4.36

CH₂), 6.97e7.71 (16H, m, phenyl, thiazoline), 10.46 and 11.34 (1H,

2s, NeH).

(2H, d, J ¼ 9 Hz, SeCH₂), 6.92e6.96 (3H, m, thiazoline, 4-

methoxyphenyl), 7.36e7.70 (12H, m, phenyl, 4-methoxyphenyl),

12.14 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 35.30 (CH₂), 102.50

(CH), 121.93 (CH), 125.05 (2CH), 125.16 (2CH), 128.30 (CH), 128.94

(2CH), 129.16 (2CH), 129.27 (CH), 129.63 (CH), 129.81 (CH), 130.26

(2CH), 130.74 (C), 133.62 (C), 139.66 (C), 140.40 (2C), 154.03 (C),

167.04 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 35.53 (CH₂), 55.94

(CH₃, OCH₃), 101.40 (CH), 114.78 (2CH), 119.68 (CH), 121.80 (C),

124.43 (2CH), 124.98 (2CH), 130.81 (3CH), 131.07 (2CH), 131.46

(2CH), 133.50 (C), 141.17 (3C), 153.95 (C), 161.23 (C), 166.88 (C).

MS (ESI)(m/z): (M^þþ 1) 516.

MS (ESI)(m/z): (M^þþ 1) 486.

For C₂₄H₁₉N₇OS₂calculated: C, 59.36; H, 3.94; N, 20.19; Found: C,

59.35; H, 3.95; N, 20.21.

For C₂₅H₂₁N₇O₂S₂calculated: C, 58.24; H, 4.11; N, 19.02; Found:

C, 58.25; H, 4.13; N, 19.01.

5.1.1.4.18. N⁰-[(4-Nitrophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(1-phenyl-1H-tetrazol-5-yl)thio]acetohydrazide (18). IR (KBr) n_max

(cm^ꢂ1): 3051.47 (Aromatic CeH stretching), 2998.42, 2865.67,

2713.80 (Aliphatic CeH stretching), 1706.42 (C]O stretching),

1601.19, 1583.64, 1567.32, 1518.32, 1493.90, 1459.06 (C]N, C]C

stretching and NeH bending), 1432.77, 1405.65, 1385.86, 1340.41

(CeH bending), 1281.46, 1236.60 (CeN stretching), 1205.51, 1179.21,

1116.79, 1107.17, 1077.76, 1059.36, 1026.94, 1012.84, 1003.01 (Aro-

matic CeH in plane bending), 986.22, 964.12, 921.26, 887.34, 868.17,

850.76, 840.40, 807.86, 761.05, 738.39, 710.65, 683.77, 656.04 (Ar-

omatic CeH out of plane bending and CeS stretching).

5.1.1.4.21. N⁰-[(4-Fluorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-phenyl-1H-tetrazol-5-yl)thio]acetohydrazide (21). IR (KBr)

n_max(cm^ꢂ1): 3042.79 (Aromatic CeH stretching), 2878.76, 2789.99

(Aliphatic CeH stretching), 1718.32 (C]O stretching), 1601.64,

1580.79, 1563.79, 1495.43, 1459.73 (C]N, C]C stretching and NeH

bending), 1433.58, 1410.86, 1392.56 (CeH bending), 1302.22,

1278.20, 1229.76 (CeN stretching), 1197.61, 1161.24, 1126.60,

1090.12, 1072.59, 1055.48, 1022.55, 1011.93 (Aromatic CeH in plane

bending), 972.60, 960.57, 919.97, 910.78, 881.99, 834.64, 824.13,

799.66, 756.33, 694.23, 685.64 (Aromatic CeH out of plane bending

and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 4.32 (2H, s, SeCH₂),

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 4.35 (2H, d, J ¼ 9 Hz, Se

7.13e7.68 (15H, m, thiazoline, phenyl, 4-nitrophenyl), 12.04 (1H, s,

NeH).

CH₂), 7.00 (1H, s, thiazoline), 7.22e7.68 (14H, m, phenyl, 4-

ﬂuorophenyl), 12.12 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d

(ppm): 35.41 (CH₂), 104.37

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.70 (CH₂), 101.58

(CH), 123.96 (CH), 124.39 (2CH), 124.88 (2CH), 130.43 (2CH), 130.78

(2CH), 130.97 (2CH), 131.40 (2CH), 133.51 (CH), 133.93 (C), 138.99

(2C), 148.67 (3C), 153.86 (C), 166.92 (C).

(CH), 115.57 (CH), 115.79 (CH), 123.36 (CH), 123.48 (2CH), 124.21

(2CH), 127.74 (C), 130.04 (2CH), 130.26 (CH), 130.67 (2CH), 130.98

(CH), 131.07 (CH), 132.74 (C), 139.35 (3C), 153.15 (C), 161.68 (C),

166.14 (C).

MS (ESI)(m/z): (M^þþ 1) 531.

For C₂₄H₁₈N₈O₃S₂calculated: C, 54.33; H, 3.42; N, 21.12; Found:

C, 54.35; H, 3.40; N, 21.11.

MS (ESI)(m/z): (M^þþ 1) 504.

For C₂₄H₁₈FN₇OS₂calculated: C, 57.24; H, 3.60; N, 19.47; Found:

C, 57.23; H, 3.59; N, 19.46.

5.1.1.4.19. N⁰-[(4-Methylphenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-phenyl-1H-tetrazol-5-yl)thio]acetohydrazide (19). IR (KBr)

n_max(cm^ꢂ1): 3054.94 (Aromatic CeH stretching), 2865.96

(Aliphatic CeH stretching), 1719.31 (C]O stretching), 1604.11,

1595.64, 1566.99, 1511.36, 1497.15, 1458.35 (C]N, C]C stretching

and NeH bending), 1430.63, 1400.19, 1388.33, 1372.96 (CeH

bending), 1335.31, 1304.67, 1276.09, 1241.89 (CeN stretching),

1198.38, 1126.98, 1091.18, 1075.17, 1059.47, 1028.44, 1012.88,

1004.22 (Aromatic CeH in plane bending), 977.13, 959.48, 917.70,

881.47, 842.12, 813.89, 793.92, 744.85, 756.30, 693.44 (Aromatic Ce

H out of plane bending and CeS stretching).

5.1.1.4.22. N⁰-[(4-Chlorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-phenyl-1H-tetrazol-5-yl)thio]acetohydrazide (22). IR (KBr)

n_max(cm^ꢂ1): 3041.64 (Aromatic CeH stretching), 2873.45 (Aliphatic

CeH stretching), 1715.75 (C]O stretching), 1611.51, 1595.09,

1570.70, 1557.71, 1489.96, 1458.35 (C]N, C]C stretching and NeH

bending), 1427.60, 1402.09, 1390.57 (CeH bending), 1300.05,

1278.84, 1241.02 (CeN stretching), 1198.92, 1125.86, 1090.33,

1073.86, 1055.84, 1025.05, 1010.90 (Aromatic CeH in plane

bending), 975.60, 959.49, 913.82, 882.48, 847.42, 830.06, 808.10,

778.74, 751.28, 709.97, 691.25, 683.47 (Aromatic CeH out of plane

bending and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 2.30 (3H, s, CH₃), 4.36

(2H, d, J ¼ 9 Hz, SeCH₂), 6.98 (1H, s, thiazoline), 7.18e7.68 (14H, m,

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 4.35 (2H, d, J ¼ 6 Hz, Se

phenyl, 4-methylphenyl), 12.16 (1H, s, NeH).

CH₂), 7.03 (1H, s, thiazoline), 7.33e7.70 (14H, m, phenyl, 4-

chlorophenyl), 12.09 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 21.64 (CH₃), 35.59

(CH₂), 102.08 (CH), 124.46 (CH), 124.73 (2CH), 124.98 (2CH), 128.81

(C), 129.10 (CH), 129.93 (2CH), 130.85 (2CH), 131.09 (2CH), 131.46

(2CH), 133.55 (2C), 140.69 (C), 141.30 (2C), 153.92 (C), 166.87 (C).

MS (ESI)(m/z): (M^þþ 1) 500.

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 35.41 (CH₂), 102.00

(CH), 124.99 (2CH), 126.77 (3CH), 129.45 (2CH), 130.82 (C), 130.88

(2CH), 130.96 (3CH), 131.45 (2CH), 133.54 (2C), 135.63 (C), 139.73

(2C), 153.96 (C), 167.01 (C).

For C₂₅H₂₁N₇OS₂calculated: C, 60.10; H, 4.24; N,19.62; Found: C,

60.12; H, 4.25; N, 19.63.

MS (ESI)(m/z): (M^þþ 1) 520.

For C₂₄H₁₈ClN₇OS₂calculated: C, 55.43; H, 3.49; N, 18.85; Found:

C, 55.42; H, 3.51; N, 18.84.

5.1.1.4.20. N⁰-[(4-Methoxyphenyl-3-phenylthiazol-2(3H)-yli-

dene)]-2-[(1-phenyl-1H-tetrazol-5-yl)thio]acetohydrazide

(20).

5.1.1.4.23. N⁰-[(4-Bromophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-phenyl-1H-tetrazol-5-yl)thio]acetohydrazide (23). IR (KBr)

n_max(cm^ꢂ1): 3053.59 (Aromatic CeH stretching), 2873.39

IR (KBr) n_max(cm^ꢂ1): 3047.43, 3003.90 (Aromatic CeH stretching),

2839.26, 2755.74 (Aliphatic CeH stretching), 1712.55 (C]O

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

273

(Aliphatic CeH stretching), 1715.31 (C]O stretching), 1611.83,

1595.08, 1570.56, 1495.96, 1457.97 (C]N, C]C stretching and NeH

bending), 1399.60 (CeH bending), 1299.59, 1279.05, 1247.77 (CeN

stretching), 1199.44, 1124.69, 1087.91, 1073.75, 1056.14, 1023.65,

1007.92 (Aromatic CeH in plane bending), 976.79, 958.57, 912.47,

882.81, 846.65, 828.60, 807.02, 751.10, 711.60, 699.15, 689.58,

682.91 (Aromatic CeH out of plane bending and CeS stretching).

909.14, 888.78, 865.71, 852.39, 837.60, 810.92, 778.23, 765.70,

740.97, 708.49, 691.18 (Aromatic CeH out of plane bending and CeS

stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 2.59 (3H, s, CH₃), 4.21

(2H, d, J ¼ 9 Hz, SeCH₂), 7.14 (1H, s, thiazoline), 7.27e7.54 (5H, m,

phenyl), 7.74 (2H, d, J ¼ 9 Hz, 4-nitrophenyl), 8.24 (2H, d, J ¼ 9 Hz, 4-

nitrophenyl), 11.99 (1H, s, NeH).

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 4.35 (2H, d, J ¼ 3 Hz, Se

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 15.71 (CH₃), 35.48

CH₂), 6.99 (1H, s, thiazoline), 7.32e7.70 (14H, m, phenyl, 4-

bromophenyl), 12.02 (1H, s, NeH).

(CH₂), 102.80 (CH), 123.63 (CH), 124.39 (2CH), 127.64 (2CH), 130.22

(2CH), 130.91 (2CH), 134.21 (C), 138.87 (2C), 148.51 (3C), 164.15 (C),

166.26 and 167.35 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 35.51 (CH₂), 103.00

(CH), 124.20 (C), 124.56 (CH), 125.00 (2CH), 125.07 (2CH), 126.90

(2CH), 129.01 (CH), 130.83 (2CH), 131.02 (C), 131.15 (2CH), 131.46

(2CH), 132.40 (C), 133.53 (C), 139.95 (2C), 153.97 (C), 166.99 (C).

MS (ESI)(m/z): (M^þþ 2) 566.

MS (ESI)(m/z): (M^þþ 1) 485.

For C₂₀H₁₆N₆O₃S₃calculated: C, 49.57; H, 3.33; N, 17.34; Found:

C, 49.55; H, 3.34; N, 17.36.

5.1.1.4.27. N⁰-[(4-Methylphenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(5-methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (27). IR (KBr)

n_max(cm^ꢂ1): 3037.33 (Aromatic CeH stretching), 2890.43 (Aliphatic

CeH stretching), 1706.73 (C]O stretching), 1602.97, 1557.64,

1509.59, 1480.58, 1455.68 (C]N, C]C stretching and NeH

bending), 1427.62, 1370.00 (CeH bending), 1259.28, 1229.94 (CeN

stretching), 1214.55, 1186.58, 1073.63, 1046.71, 1030.98, 1017.64,

1002.56 (Aromatic CeH in plane bending), 955.19, 880.59, 839.81,

819.18, 808.22, 781.67, 759.38, 739.61, 694.39 (Aromatic CeH out of

plane bending and CeS stretching).

For C₂₄H₁₈BrN₇OS₂calculated: C, 51.07; H, 3.21; N, 17.37; Found:

C, 51.05; H, 3.20; N, 17.36.

5.1.1.4.24. N⁰-[(4-Cyanophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(1-phenyl-1H-tetrazol-5-yl)thio]acetohydrazide (24). IR (KBr)

n_max(cm^ꢂ1): 3438.84 (NeH stretching), 3053.11 (Aromatic CeH

stretching), 2864.09, 2752.23 (Aliphatic CeH stretching), 2227.63

(C^N stretching), 1708.81 (C]O stretching), 1602.74, 1577.66,

1496.66 (C]N, C]C stretching and NeH bending), 1406.01 (CeH

bending), 1284.50 (CeN stretching), 1116.71 (Aromatic CeH in

plane bending), 854.41, 759.90, 694.33 (Aromatic CeH out of plane

bending and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 2.34 (3H, s, CH₃), 2.66

(3H, s, CH₃, thiadiazole), 4.23 (2H, s, SeCH₂), 6.98 (1H, s, thiazoline),

7.22e7.59 (9H, m, phenyl, 4-methylphenyl), 12.11 (1H, s, NeH).

¹H NMR (400 MHz) (DMSO-d₆)

d

(ppm): 4.42 (2H, d, J ¼ 10.4 Hz,

SeCH₂), 7.28 (1H, s, thiazoline), 7.40e7.43 (3H, m, phenyl), 7.55e

7.59 (5H, m, phenyl), 7.64e7.73 (4H, m, phenyl, 4-cyanophenyl),

7.90 (2H, d, J ¼ 8.4 Hz, 4-cyanophenyl), 12.37 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 15.91 (CH₃), 21.65

(CH₃), 35.93 (CH₂), 102.05 (CH), 124.48 (3CH), 124.72 (C), 128.84

(2CH),129.11 (2CH), 129.96 (2CH), 131.08 (C), 140.66 (C), 141.37 (3C),

164.47 (C), 166.42 and 167.15 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 34.73 (CH₂), 103.48

(CH), 112.52 (C), 118.21 (C), 123.31 (CH), 124.18 (2CH), 127.58 (2CH),

129.11 (2CH),129.70 (CH),130.04 (2CH),130.22 (2CH),130.66 (2CH),

132.44 (C), 132.74 (C), 138.60 (3C), 153.11 (C), 166.14 (C).

MS (ESI)(m/z): (M^þþ 1) 511.

MS (ESI)(m/z): (M^þþ 1) 454.

For C₂₁H₁₉N₅OS₃calculated: C, 55.60; H, 4.22; N,15.44; Found: C,

55.61; H, 4.20; N, 15.45.

5.1.1.4.28. N⁰-[(4-Methoxyphenyl-3-phenylthiazol-2(3H)-yli-

dene)]-2-[(5-methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (28).

IR (KBr) n_max(cm^ꢂ1): 2998.70, 2901.02 (Aliphatic CeH stretching),

1717.46 (C]O stretching), 1602.24, 1560.36, 1509.10, 1489.85,

1455.92 (C]N, C]C stretching and NeH bending), 1424.03,

1397.60, 1372.00, 1354.23 (CeH bending), 1294.48, 1251.05, 1226.20

(CeN stretching), 1182.55, 1133.69, 1064.62, 1035.52, 1020.08 (Ar-

omatic CeH in plane bending), 955.01, 884.79, 852.09, 838.10,

822.02, 783.55, 762.97, 743.91, 695.88 (Aromatic CeH out of plane

bending and CeS stretching).

For C₂₅H₁₈N₈OS₂calculated: C, 58.81; H, 3.55; N, 21.95; Found: C,

58.82; H, 3.52; N, 21.97.

5.1.1.4.25. N⁰-[(3,4-Diphenylthiazol-2(3H)-ylidene)]-2-[(5-

methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (25) [36]. IR (KBr)

n_max(cm^ꢂ1): 3089.35 (Aromatic CeH stretching), 2867.03 (Aliphatic

CeH stretching), 1706.41 (C]O stretching), 1601.98, 1581.41,

1518.32, 1490.97 (C]N, C]C stretching and NeH bending),

1443.60, 1385.33, 1342.49 (CeH bending), 1282.24, 1236.05 (CeN

stretching), 1195.45, 1153.90, 1116.47, 1076.69, 1026.93, 1012.08

(Aromatic CeH in plane bending), 959.19, 921.19, 888.11, 868.06,

850.63, 807.40, 761.50, 738.33, 690.85 (Aromatic CeH out of plane

bending and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 2.65 (3H, s, CH₃), 3.80

(3H, s, OCH₃), 4.24 (2H, s, SeCH₂), 6.96e6.99 (3H, m, thiazoline, 4-

methoxyphenyl), 7.39e7.59 (7H, m, phenyl, 4-methoxyphenyl),

12.13 (1H, s, NeH).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 2.61 and 2.68 (3H, 2s,

CH₃), 4.05e4.12 (2H, m, SeCH₂), 6.96e7.43 (11H, m, thiazoline,

phenyl), 10.36 and 11.29 (1H, 2s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 15.84 (CH₃), 36.04

(CH₂), 56.04 (CH₃), 101.66 (CH), 114.85 (2CH), 119.71 (CH), 124.55

(C),128.93 (2CH),130.88 (2CH),131.10 (2CH),138.79 (C), 141.30 (3C),

161.31 (C), 164.49 (C), 166.49 and 167.15 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 15.96 (CH₃), 36.09

(CH₂), 102.84 (CH), 124.51 (3CH), 127.54 (CH), 129.29 (2CH), 129.43

(2CH), 130.89 (2CH), 131.09 (C), 138.88 (C), 141.29 (3C), 164.45 (C),

166.52 and 167.14 (C).

MS (ESI)(m/z): (M^þþ 1) 470.

For C₂₁H₁₉N₅O₂S₃calculated: C, 53.71; H, 4.08; N, 14.91; Found :

C, 53.70; H, 4.10; N, 14.92.

MS (ESI)(m/z): (M^þþ 1) 440.

For C₂₀H₁₇N₅OS₃calculated: C, 54.65; H, 3.90; N,15.93; Found: C,

54.66; H, 3.92; N, 15.90.

5.1.1.4.29. N⁰-[(4-Fluorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(5-methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (29). IR (KBr)

n_max(cm^ꢂ1): 3058.84 (Aromatic CeH stretching), 2899.54

(Aliphatic CeH stretching), 1708.38 (C]O stretching), 1601.57,

1582.91, 1563.21, 1506.61, 1493.66, 1450.70 (C]N, C]C stretching

and NeH bending), 1384.77, 1351.30 (CeH bending), 1225.19 (CeN

stretching), 1185.93, 1163.54, 1103.92, 1070.29, 1026.05 (Aromatic

CeH in plane bending), 883.45, 856.84, 821.97, 791.26, 777.42,

746.95, 694.88 (Aromatic CeH out of plane bending and CeS

stretching).

5.1.1.4.26. N⁰-[(4-Nitrophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(5-methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (26). IR (KBr)

n_max(cm^ꢂ1): 3068.23 (Aromatic CeH stretching), 2902.74 (Aliphatic

CeH stretching), 1725.01 (C]O stretching), 1603.82, 1566.58,

1513.90, 1495.24, 1475.36, 1459.37 (C]N, C]C stretching and NeH

bending), 1409.26, 1381.52, 1346.75 (CeH bending), 1291.94,

1232.17 (CeN stretching), 1191.79, 1165.64, 1121.52, 1108.97,

1069.62,1022.03,1013.07 (Aromatic CeH in plane bending), 954.42,

274

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 2.65 (3H, s, CH₃), 4.22

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 15.13 (CH₃), 34.92

(2H, s, SeCH₂), 7.00 (1H, s, thiazoline), 7.24e7.57 (9H, m, phenyl, 4-

ﬂuorophenyl), 12.04 (1H, s, NeH).

(CH₂), 103.47 (CH), 112.37 (C), 118.24 (C), 120.80 (CH), 123.32 (2CH),

127.61 (2CH), 129.09 (2CH), 130.23 (2CH), 132.44 (C), 138.66 (4C),

163.40 (C), 165.57 and 166.50 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 15.83 (CH₃), 35.89

(CH₂), 102.82 (CH), 116.36 (CH), 116.58 (CH), 121.75 (CH), 124.39

(2CH), 128.75 (C), 130.48 (2CH), 131.06 (CH), 131.91 (CH), 137.88 (C),

140.28 (3C), 162.50 (C), 164.29 (C), 166.44 and 167.21 (C).

MS (ESI)(m/z): (M^þþ 1) 458.

MS (ESI)(m/z): (M^þþ 1) 465.

For C₂₁H₁₆N₆OS₃calculated: C, 54.29; H, 3.47; N,18.09; Found: C,

54.32; H, 3.45; N, 18.11.

5.1.1.4.33. N⁰-[(3,4-Diphenylthiazol-2(3H)-ylidene)]-2-[(pyr-

imidin-2-yl)thio]acetohydrazide (33). IR (KBr) n_max(cm^ꢂ1): 3116.21

(Aromatic CeH stretching), 2927.91 (Aliphatic CeH stretching),

1673.26 (C]O stretching), 1618.49, 1583.35, 1550.63, 1489.09 (C]

N, C]C stretching and NeH bending), 1445.64, 1379.64 (CeH

bending), 1304.73, 1239.08 (CeN stretching), 1172.91, 1072.89,

1014.78 (Aromatic CeH in plane bending), 958.97, 925.10, 881.62,

829.55, 806.50, 766.78, 747.03, 727.95, 693.79 (Aromatic CeH out of

plane bending and CeS stretching).

For C₂₀H₁₆FN₅OS₃calculated: C, 52.50; H, 3.52; N, 15.31; Found:

C, 52.52; H, 3.51; N, 15.29.

5.1.1.4.30. N⁰-[(4-Chlorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(5-methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (30). IR (KBr)

n_max(cm^ꢂ1): 3468.63 (NeH stretching), 3056.87 (Aromatic CeH

stretching), 2962.51, 2918.10 (Aliphatic CeH stretching), 1708.68

(C]O stretching), 1600.34, 1567.72, 1557.93, 1489.71, 1451.18 (C]N,

C]C stretching and NeH bending), 1428.07, 1406.25, 1383.78,

1350.71 (CeH bending), 1225.28 (CeN stretching), 1186.78, 1155.33,

1087.85, 1070.70, 1052.86, 1026.74, 1010.63 (Aromatic CeH in plane

bending), 949.91, 918.06, 883.88, 854.58, 832.85, 806.38, 779.61,

759.95, 749.89, 707.23, 694.01 (Aromatic CeH out of plane bending

and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.91 and 3.94 (2H, 2s,

SeCH₂), 6.98e7.44 (12H, m, thiazoline, phenyl, pyrimidine), 8.48

(1H, d, J ¼ 6 Hz, pyrimidine), 8.66 (1H, d, J ¼ 3 Hz, pyrimidine),10.30

and 11.16 (1H, 2s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 33.03 (CH₂), 101.00

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 2.67 (3H, s, CH₃), 4.23

(CH), 118.09 (CH), 121.61 (CH), 121.88 (2CH), 128.28 (CH), 128.94

(CH), 128.99 (CH), 129.26 (2CH), 129.63 (CH), 129.68 (CH), 130.25

(C), 139.95 (C), 140.38 (2C), 158.34 (CH), 158.50 (CH), 168.16 (C),

170.20 (C).

(2H, s, SeCH₂), 7.03 (1H, s, thiazoline), 7.34e7.57 (9H, m, phenyl, 4-

chlorophenyl), 12.04 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 15.89 (CH₃), 35.69

(CH₂), 102.81 (CH), 121.53 (2CH), 124.07 (CH), 126.51 (2CH), 128.43

(C), 129.46 (2CH), 130.42 (2CH), 131.02 (C), 135.65 (C), 139.90 (3C),

164.29 (C), 166.41 and 167.25 (C).

MS (ESI)(m/z): (M^þþ 1) 420.

For C₂₁H₁₇N₅OS₂calculated: C, 60.12; H, 4.08; N, 16.69; Found: C,

60.10; H, 4.11; N, 16.71.

MS (ESI)(m/z): (M^þþ 1) 474.

5.1.1.4.34. N⁰-[(4-Nitrophenyl-3-phenylthiazol-2(3H)-ylidene)]-2-

[(pyrimidin-2-yl)thio]acetohydrazide (34). IR (KBr) n_max(cm^ꢂ1):

3099.76, 3048.66 (Aromatic CeH stretching), 2916.81 (Aliphatic Ce

H stretching), 1720.72 (C]O stretching), 1602.77, 1588.89, 1571.48,

1552.18, 1513.61, 1491.23, 1455.61 (C]N, C]C stretching and NeH

bending), 1445.73, 1351.48 (CeH bending), 1318.33, 1287.98,

1253.72 (CeN stretching), 1195.34, 1173.10, 1158.16, 1116.35,

1075.68, 1026.26, 1002.42 (Aromatic CeH in plane bending),

920.62, 866.11, 852.66, 810.07, 764.91, 739.15, 691.88 (Aromatic Ce

H out of plane bending and CeS stretching).

For C₂₀H₁₆ClN₅OS₃calculated: C, 50.68; H, 3.40; N, 14.77; Found:

C, 50.69; H, 3.42; N, 14.76.

5.1.1.4.31. N⁰-[(4-Bromophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(5-methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (31). IR (KBr)

n_max(cm^ꢂ1): 3061.03 (Aromatic CeH stretching), 2882.43, 2750.05

(Aliphatic CeH stretching), 1716.93 (C]O stretching), 1597.84,

1587.35, 1571.34, 1555.38, 1508.37, 1482.05, 1453.65 (C]N, C]C

stretching and NeH bending), 1421.97, 1396.34, 1379.30 (CeH

bending), 1260.27 (CeN stretching), 1217.93, 1186.32, 1127.90,

1073.22, 1060.82, 1031.15, 1007.78 (Aromatic CeH in plane

bending), 970.35, 957.25, 925.19, 878.29, 852.92, 830.73, 810.64,

772.15, 755.12, 701.88, 670.49 (Aromatic CeH out of plane bending

and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.93 and 4.08 (2H, 2d, J₁,

J₂¼ 15 Hz, SeCH₂), 7.11e7.56 (7H, m, thiazoline, phenyl, pyrimi-

dine), 7.70 (2H, d, J ¼ 9 Hz, 4-nitrophenyl), 8.08 (2H, d, J ¼ 9 Hz, 4-

nitrophenyl), 8.45 (2H, d, J ¼ 6 Hz, pyrimidine), 11.77 (1H, s, NeH).

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 2.68 (3H, s, CH₃), 4.24

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 33.20 (CH₂), 104.77

(2H, s, SeCH₂), 7.05 (1H, s, thiazoline), 7.34e7.64 (9H, m, phenyl, 4-

bromophenyl), 12.04 (1H, s, NeH).

(CH), 118.10 (CH), 124.06 (3CH), 124.14 (2CH), 128.48 (2CH), 130.51

(CH), 131.08 (CH), 133.97 (C), 139.24 (C), 148.47 (3C), 158.36 (2CH),

168.27 (C), 169.83 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 16.06 (CH₃), 35.89

(CH₂), 103.31 (CH), 124.43 (C), 124.64 (CH), 126.73 (2CH), 128.81

(2CH), 131.09 (C), 131.25 (2CH), 132.19 (2CH), 132.43 (C), 140.15 (3C),

164.34 (C), 166.44 and 167.28 (C).

MS (ESI)(m/z): (M^þþ 1) 465.

For C₂₁H₁₆N₆O₃S₂calculated: C, 54.30; H, 3.47; N, 18.09; Found:

C, 54.32; H, 3.46; N, 18.11.

MS (ESI)(m/z): (M^þþ 1) 519.

5.1.1.4.35. N⁰-[(4-Methylphenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(pyrimidin-2-yl)thio]acetohydrazide (35). IR (KBr) n_max(cm^ꢂ1):

3178.95 (NeH stretching), 1678.05 (C]O stretching), 1609.67,

1584.42, 1553.13, 1489.35 (C]N, C]C stretching and NeH

bending), 1446.33, 1380.68 (CeH bending), 1307.59, 1235.57 (CeN

stretching), 1183.19, 1070.10, 1027.89, 1015.99 (Aromatic CeH in

plane bending), 956.22, 883.18, 811.97, 765.62, 746.57, 695.04 (Ar-

omatic CeH out of plane bending and CeS stretching).

For C₂₀H₁₆BrN₅OS₃calculated: C, 46.33; H, 3.11; N, 13.51; Found:

C, 46.34; H, 3.12; N, 13.50.

5.1.1.4.32. N⁰-[(4-Cyanophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(5-methyl-1,3,4-thiadiazol-2-yl)thio]acetohydrazide (32). IR (KBr)

n_max(cm^ꢂ1): 3444.63 (NeH stretching), 3045.39 (Aromatic CeH

stretching), 2902.67 (Aliphatic CeH stretching), 2223.77 (C^N

stretching), 1724.24 (C]O stretching), 1610.45, 1569.95, 1477.37

(C]N, C]C stretching and NeH bending), 1382.87 (CeH bending),

1292.22 (CeN stretching), 1191.93, 1122.49 (Aromatic CeH in plane

bending), 833.19, 757.97 (Aromatic CeH out of plane bending).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 2.30 (3H, s, CH₃), 3.93

(2H, s, SeCH₂), 6.34 (1H, phenyl), 6.97e7.39 (10H, m, thiazoline,

phenyl, 4-methylphenyl, pyrimidine), 8.49 (2H, d, J ¼ 3 Hz, py-

rimidine), 11.09 (1H, s, NeH).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 2.69 (3H, s, CH₃), 4.29

(2H, d, J ¼ 5.2 Hz, SeCH₂), 7.27 (1H, s, thiazoline), 7.42 (3H, d,

J ¼ 7.2 Hz, phenyl), 7.57 (2H, t, J₁¼ 7.6 Hz, J₂¼ 8 Hz, phenyl), 7.70

(2H, d, J ¼ 8.4 Hz, 4-cyanophenyl), 7.88e7.90 (2H, d, J ¼ 8 Hz, 4-

cyanophenyl), 11.51 and 12.28 (1H, 2s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 21.55 (CH₃), 33.02

(CH₂), 95.00 (CH), 118.06 (CH), 121.96 (3CH), 128.21 (C), 129.55

(2CH), 129.69 (2CH), 130.29 (2CH), 139.24 (2C), 140.05 (2C), 158.31

(2CH), 168.15 (C), 170.18 (C).

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

275

MS (ESI)(m/z): (M^þþ 1) 434.

¹³C NMR (100 MHz) (DMSO-d₆)

d

(ppm): 32.26 (CH₂), 103.32

For C₂₂H₁₉N₅OS₂calculated: C, 60.95; H, 4.42; N, 16.15; Found: C,

60.95; H, 4.40; N, 16.16.

(CH), 118.23 (CH), 124.46 (2CH), 126.37 (CH), 128.90 (2CH), 129.18

(C), 131.05 (2CH), 131.11 (2CH), 135.67 (C), 138.96 (C), 140.34 (2C),

158.34 (2CH), 168.14 (C), 169.75 (C).

5.1.1.4.36. N⁰-[(4-Methoxyphenyl-3-phenylthiazol-2(3H)-yli-

dene)]-2-[(pyrimidin-2-yl)thio]acetohydrazide (36). IR (KBr) n_max

(cm^ꢂ1): 3458.23 (NeH stretching), 3090.59, 3061.82, 3006.60 (Ar-

omatic CeH stretching), 2906.63 (Aliphatic CeH asymmetric

stretching), 2839.64 (Aliphatic CeH symmetric stretching), 1707.44

(C]O stretching), 1603.05, 1571.51, 1548.99, 1509.43, 1495.36,

1455.61 (C]N, C]C stretching and NeH bending), 1426.26,

1381.49, 1363.53 (CeH bending), 1309.03, 1296.15, 1249.38, 1228.42

(CeN stretching), 1204.89 (CeO stretching), 1180.18, 1147.29,

1070.66, 1049.92, 1037.73, 1070.66, 1021.86 (Aromatic CeH in plane

bending), 919.16, 884.81, 847.85, 831.51, 814.28, 781.39, 756.20,

736.26, 718.95, 693.25 (Aromatic CeH out of plane bending and Ce

S stretching).

MS (ESI)(m/z): (M^þþ 1) 454.

For C₂₁H₁₆ClN₅OS₂calculated: C, 55.56; H, 3.55; N, 15.43; Found:

C, 55.55; H, 3.55; N, 15.44.

5.1.1.4.39. N⁰-[(4-Bromophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(pyrimidin-2-yl)thio]acetohydrazide (39). IR (KBr) n_max(cm^ꢂ1):

3448.17, 3222.00 (NeH stretching), 3104.75 (Aromatic CeH

stretching), 2896.91 (Aliphatic CeH stretching), 1711.55 (C]O

stretching), 1606.93, 1586.30, 1570.01, 1552.24, 1494.72, 1470.70,

1450.75 (C]N, C]C stretching and NeH bending), 1423.41,

1402.49, 1378.75 (CeH bending), 1267.03, 1232.97 (CeN stretch-

ing), 1183.85, 1165.79, 1141.66, 1074.38, 1053.64, 1024.00, 1008.74

(Aromatic CeH in plane bending), 906.46, 877.71, 823.54, 811.65,

783.10, 772.48, 754.67, 744.71, 702.92, 688.55 (Aromatic CeH out of

plane bending and CeS stretching).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.77 (3H, s, OCH₃), 4.01

and 4.13 (2H, 2d, J₁, J₂¼ 15 Hz, SeCH₂), 6.81e6.84 (3H, m, phenyl, 4-

methoxyphenyl), 6.89 (1H, s, thiazoline), 7.18e7.58 (7H, m, phenyl,

4-methoxyphenyl, pyrimidine), 8.49 (2H, d, J ¼ 6 Hz, pyrimidine),

11.97 (1H, s, NeH).

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.95 and 4.09 (2H, 2d, J₁,

J₂¼ 15 Hz, SeCH₂), 7.02 (1H, s, thiazoline), 7.20e7.58 (10H, m,

phenyl, 4-bromophenyl, pyrimidine), 8.51 (2H, d, J ¼ 3 Hz, pyrim-

idine), 11.76 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 33.29 (CH₂), 55.97

(CH₃), 102.00 (CH), 114.52 (2CH), 118.18 (2CH), 119.62 (C), 124.61

(2CH), 130.81 (2CH), 131.12 (2CH), 138.45 (C), 141.57 (2C), 158.37

(2CH), 161.08 (C), 168.07 (C), 169.83 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 33.27 (CH₂), 103.19

(CH), 118.26 (CH), 124.45 (C), 124.52 (CH), 126.80 (2CH), 128.86

(2CH), 131.12 (C), 131.22 (2CH), 132.11 (2CH), 140.40 (3C), 158.37

(2CH), 168.17 (C), 169.77 (C).

MS (ESI)(m/z): (M^þþ 1) 450.

For C₂₂H₁₉N₅O₂S₂calculated: C, 58.78; H, 4.26; N, 15.58; Found:

C, 58.80; H, 4.25; N, 15.58.

MS (ESI)(m/z): (M^þþ 1) 499.

For C₂₁H₁₆BrN₅OS₂calculated: C, 50.60; H, 3.24; N,14.05; Found:

C, 50.60; H, 3.25; N, 14.02.

5.1.1.4.37. N⁰-[(4-Fluorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(pyrimidin-2-yl)thio]acetohydrazide (37). IR (KBr) n_max(cm^ꢂ1):

3439.55, 3214.83 (NeH stretching), 3111.10 (Aromatic CeH

stretching), 2897.96 (Aliphatic CeH stretching), 1710.06 (C]O

stretching), 1605.03, 1582.01, 1565.56, 1548.99, 1507.25, 1496.78,

1468.84 (C]N, C]C stretching and NeH bending), 1449.85,

1413.21, 1379.79, 1364.77 (CeH bending), 1252.01, 1232.50 (CeN

stretching), 1204.74, 1190.26, 1165.02, 1147.37, 1106.85, 1067.77,

1051.30, 1024.36, 1012.48 (Aromatic CeH in plane bending), 908.86,

879.03, 843.44, 831.39, 808.43, 792.62, 772.88, 754.16, 745.95,

707.60 (Aromatic CeH out of plane bending).

5.1.1.4.40. N⁰-[(4-Cyanophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(pyrimidin-2-yl)thio]acetohydrazide (40). IR (KBr) n_max(cm^ꢂ1):

3419.56 (NeH stretching), 3085.89 (Aromatic CeH stretching),

2912.31, 2827.45 (Aliphatic CeH stretching), 2229.56 (C^N

stretching), 1720.39 (C]O stretching), 1604.66, 1562.23, 1548.73,

1477.37 (C]N, C]C stretching and NeH bending), 1390.58 (CeH

bending), 1255.57 (CeN stretching), 1182.28, 1122.49, 1033.77 (Ar-

omatic CeH in plane bending), 802.33, 765.69, 628.75 (Aromatic Ce

H out of plane bending and CeS stretching).

¹H NMR (400 MHz) (DMSO-d₆)

d (ppm): 3.99 and 4.16 (2H, 2d, J₁,

¹H NMR (300 MHz) (DMSO-d₆)

d

(ppm): 3.96 and 4.09 (2H, 2d,

J₂¼ 11.7 Hz, SeCH₂), 7.23 (1H, t, J₁¼ 5.2 Hz, J₂¼ 4.4 Hz, pyrimidine),

7.27 (1H, s, thiazoline), 7.41e7.45 (3H, m, phenyl), 7.58 (2H, t,

J₁¼ 7.2 Hz, J₂¼ 8.4 Hz, phenyl), 7.66 (2H, d, J ¼ 8 Hz, 4-

cyanophenyl), 7.76 (2H, d, J ¼ 8.4 Hz, 4-cyanophenyl), 8.52 (2H, d,

J ¼ 4.8 Hz, pyrimidine), 11.29 and 11.99 (1H, 2s, NeH).

J₁¼ 15 Hz, J₂¼ 18 Hz, SeCH₂), 7.01 (1H, s, thiazoline), 7.09e7.59

(10H, m, phenyl, 4-ﬂuorophenyl, pyrimidine), 8.52 (2H, d, J ¼ 3 Hz,

pyrimidine), 11.79 (1H, s, NeH).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 33.20 (CH₂), 102.88

(CH), 116.08 (CH), 116.30 (CH), 118.24 (CH), 123.99 (CH), 124.49

(2CH),128.91 (C),131.12 (2CH),131.81 (CH),131.89 (CH),140.52 (3C),

158.42 (2CH), 162.31 (C), 168.13 (C), 169.79 (C).

¹³C NMR (100 MHz) (DMSO-d₆)

d (ppm): 32.43 (CH₂), 103.61

(CH), 112.37 (C), 117.46 (CH), 118.06 (C), 120.84 (CH), 123.42 (2CH),

127.78 (2CH), 129.16 (2CH), 130.28 (2CH), 132.16 (C), 138.87 (3C),

157.62 (2CH), 167.41 (C), 169.03 (C).

MS (ESI)(m/z): (M^þþ 1) 438.

For C₂₁H₁₆FN₅OS₂calculated: C, 57.65; H, 3.69; N, 16.01; Found:

C, 57.65; H, 3.71; N, 16.02.

MS (ESI)(m/z): (M^þþ 1) 445.

For C₂₂H₁₆N₆OS₂calculated: C, 59.44; H, 3.63; N,18.91; Found: C,

59.43; H, 3.61; N, 18.90.

5.1.1.4.38. N⁰-[(4-Chlorophenyl-3-phenylthiazol-2(3H)-ylidene)]-

2-[(pyrimidin-2-yl)thio]acetohydrazide (38). IR (KBr) n_max(cm^ꢂ1):

3446.68, 3221.62 (NeH stretching), 3106.47 (Aromatic CeH

stretching), 2896.97 (Aliphatic CeH stretching), 1712.54 (C]O

stretching), 1608.42,1571.18, 1551.32, 1492.20, 1470.47, 1450.21 (C]

N, C]C stretching and NeH bending), 1422.93, 1406.31, 1378.72,

1365.82 (CeH bending),1266.95,1240.30 (CeN stretching),1186.87,

1166.73, 1143.08, 1094.72, 1071.21, 1053.18, 1024.40, 1011.91 (Aro-

matic CeH in plane bending), 907.41, 878.07, 832.54, 810.88, 784.21,

772.79, 755.25, 744.55, 712.70, 692.71 (Aromatic CeH out of plane

bending and CeS stretching).

5.2. Microbiology

5.2.1. Antibacterial and anticandidal activity

The antimicrobial activities of compounds 1e40 were tested

using the microbroth dilution method [39]. Tested microorganism

strains were Micrococcus luteus (NRLL B-4375), Bacillus subtilis

(NRS-744), P. aeruginosa (ATCC-254992), Staphylococcus aureus

(NRRL B-767), Escherichia coli (ATCC-25922), Listeria monocytogenes

(ATCC-7644) C. albicans (ATCC-22019). Microbroth dilution-

susceptibility assay was used for antimicrobial evaluation of the

compounds. Stock solutions of the samples were prepared in

dimethyl sulfoxide. Dilution series using sterile distilled water were

prepared from 4 mg/mL to 0.0039 mg/mL in micro-test tubes that

¹H NMR (300 MHz) (DMSO-d₆)

d (ppm): 3.95 and 4.09 (2H, 2d, J₁,

J₂¼ 15 Hz, SeCH₂), 7.03 (1H, s, thiazoline), 7.19e7.58 (10H, m,

phenyl, 4-chlorophenyl, pyrimidine), 8.50 (2H, d, J ¼ 3 Hz, pyrim-

idine), 11.78 (1H, s, NeH).

276

M.D. Altıntop et al. / European Journal of Medicinal Chemistry 74 (2014) 264e277

were transferred to 96-well microtiter plates. Overnight-grown

bacterial and C. albicans suspensions in double-strength Mueller-

Hinton broth were standardized to 10⁸CFU/mL using McFarland

No: 0.5 standard solutions. Hundred microliter of each microor-

ganism suspension was then added into the wells. The last well-

chain without a microorganism was used as a negative control.

Sterile distilled water and the medium served as a positive growth

control. After incubation at 37 ^ꢁC for 18e24 h, antimicrobial activity

was detected by spraying of 0.5% TTC (triphenyl tetrazolium chlo-

ride, Merck) aqueous solution. MIC was deﬁned as the lowest

concentration of compounds that inhibited visible growth, as

indicated by the TTC staining.

removed, the formazan crystals formed by MTT metabolism were

solubilized by addition of 200 L DMSO to each well and absor-

bance was read at 540 nm with a microtitre plate spectropho-

tometer (Bio-Tek plate reader). Each concentration was repeated in

three wells and IC₅₀values were deﬁned as the drug concentrations

that reduced absorbance to 50% of control values.

m

5.3.3. Analysis of DNA synthesis

Analysis of DNA synthesis was measured by BrdU Cell Prolifer-

ation ELISA (colorimetric) kit (Roche). This immunostaining pro-

cedure is based on the use of speciﬁc anti-BrdU antibodies for

measuring the incorporation of bromodeoxyuridine (BrdU) into

nuclear DNA in place of thymidine during the S-phase of the cell

cycle [43]. Hence, this method provides a colorimetric measure-

ment for DNA synthesis inhibition ratio of the carcinogenic cells.

After cells were seeded into 96 well ﬂat-bottomed microtiter plates

at a density of 2 ꢃ 10³, various doses of compounds which were

chosen after MTT test (compounds 16, 17, 19, 21, 23, 27, 28, 29, 30)

and cisplatin were administrated into C6 tumor cells. Microtiter

plates were incubated at 37 ^ꢁC in a 5% CO₂/95% air humidiﬁed at-

5.2.2. Antifungal activity

The antifungal activities of compounds 1e40 were tested using

the microbroth dilution method with some modiﬁcations [39,40].

Tested fungal strains were A. parasiticus (NRRL-465), A. ochraceus

(NRRL 3174) Penicillium chrysogenum (NRRL 1951), Trichoderma

harzianum (NRRL 20565), F. solani (NRRL-13414), F. culmorum (wild

culture) and F. moniliforme (NRRL 1866). Microbroth dilution-

susceptibility assay was used for antimicrobial evaluation of the

compounds. Stock solutions of the samples were prepared in

dimethyl sulfoxide. Dilution series using sterile distilled water were

prepared from 4 mg/mL to 0.0039 mg/mL in micro-test tubes that

were transferred to 96-well microtiter plates. Fungal strains grown

on PDA at 25 ^ꢁC for 5 suspensions in double-strength Potato

Dextrose Broth (PDB) were standardized to 10⁵spores/mL. Hun-

dred microliter of each spore suspension was then added into the

wells. The last well-chain without a fungus was used as a negative

control. Sterile distilled water and the medium served as a positive

growth control. After incubation at 25 ^ꢁC for 48e72 h, antifungal

activity was detected by investigation of mycelia growing under

stereo microscope.

mosphere for 24 h. The cells were labeled with 10

mL BrdU solution

for 2 h and then ﬁxed. Anti-BrdU-POD (100 L) was added and

m

incubated for 90 min. After washing steps with 1xPBS, cells were

incubated with substrate. Absorbance of the samples was measured

with an ELX808-IU Bio-Tek apparatus at 450 nm. For each com-

pound dose, triplicate wells were used.

5.3.4. Statistical analyses

Statistical analyses were evaluated by Statistical Package for the

Social Science (SPSS) for Windows 15.0. Data was expressed as

Mean ꢀ SD. Comparisons were performed by one way ANOVA test

for normally distributed continuous variables and Tukey test was

used for post hoc analyses of group differences.

Streptomycin was used as standard antibacterial agent, whereas

ketoconazole was used as an antifungal agent.

Acknowledgments

5.3. Biochemistry

This study was supported by Anadolu University Scientiﬁc

Research Projects Commission under the grant no: 1105S087.

5.3.1. Cell culture and drug treatment

C6 rat glioma cells and NIH/3T3 mouse embryonic ﬁbroblast cells

were obtained from the American Type Culture Collection (ATCC,

USA). The cells were incubated in Dulbecco’s modiﬁed Eagle’s me-

dium (DMEM) supplemented with 10% fetal calf serum (Life Tech-

nologies, UK), 100 IU/mL penicillin (Gibco, Paisley, Scotland) and

100 mg/mL streptomycin (Gibco) at 37 ^ꢁC in a humidiﬁed atmo-

sphere of 95% air and 5% CO₂. Exponentially growing cells were

plated at 2 ꢃ 10⁴cells/mL into 96-well microtiter tissue culture

plates (Nunc, Denmark) and incubated for 24 h before the addition

of the drugs (the optimum cell number for cytotoxicity assays was

determined in preliminary experiments). Stock solutions of com-

pounds were prepared in dimethyl sulfoxide (DMSO; Sigmae

Aldrich, Poole, UK) and further dilutions were made with fresh

culture medium (the concentration of DMSO in the ﬁnal culture

medium was <0.1% which had no effect on the cell viability).

Appendix A. Supplementary data

Supplementary data related to this article can be found at http://

dx.doi.org/10.1016/j.ejmech.2013.12.060.

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Article Doi

Synthesis and biological evaluation of thiazoline derivatives as new antimicrobial and anticancer agents

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