Synthesis and spectral studies of 1,N-acetyl-3-aryl-5-(1’,N-phenyl-3’-o-methoxyphenyl-pyrazole-4’-yl)-pyrazoline  derivatives and their expected biological activity

 

Mrs. Kokila Parmar*, Kamlesh Kundariya , Sarju Prajapati and  Rinku Patel

Department of Chemistry Hem. North Gujarat University, Patan-384 265 Gujarat, India

*Corresponding Author E-mail: drkap_chem@yahoo.com

 

ABSTRACT:

The synthesis and spectroscopic Characterization of   1, N-Acetyl- 3 Aryl-5-(1’-N-Phenyl-3’-o-Methoxy-Phenyl Pyrazol-4’-yl)-Pyrazolines are reported in this paper. o-Methoxy Phenyl Hydrezone has been used as the Starting material in preparing all compounds. The compounds 1, N-Acetyl- 3 Aryl-5-(1’-N-Phenyl-3’-o-Methoxy-Phenyl Pyrazol-4’-yl)-Pyrazolines were characterized on the basis of elemental analysis and spectral studies. All the compounds have been screened for their in vitro biological assay like antibacterial activity towards gram positive and gram negative bacterial Strains and their antifungal Potential

 

KEYWORDS: Synthesis,  Spectral studies, Biological activity, Pyrazole,  Pyrazoline

 


 

INTRODUCTION:

Heterocyclic compounds show vital role in the field of pharmaceuticals. Pyrazole derivatives exhibit broad spectrum of pharmacological and biological activities (1-10). They have been used as anticancer, antitumor, antiulcer, antimicrobial, antiparasitic, CNS depressant, herbicidal, immunosuppressant, lipoxygenase inhibitor, neurotonsin receptor etc. Carbon Romuald and co-workers (11) have reported Pyrazole derivatives useful as reverse  transcriptase inhibitors for the treatment of HIV infection. Recently Bantwal Holla and co-workers (12) have synthesized pyrazole derivatives as potential antimicrobial agents. Anti-malarial activity of 4-(5-trifluoro methyl-1H-Pyrazole-1-yl)-chloroquine analogues has been evaluated in vitro against a chloroquine resistant plasmodium falciparum clone by Antoniana U. Krettli et.al. (13).

 

The Chemistry of Pyrazolines have been studied extensively for their biodynamic behavior (14) and industrial applications (15) It was revealed that 2- pyrazolines are better therapeutic agents (16-34). They have been used as antiallegic, anticonvalsant, antiimplantation, anti-inflammatory, antitumor, antineoplastic, antimicrobial, analgesic, bactericidal, cardiovascular, diuretic, fungicidal, herbicidal, hypoglycemic, insecticidal, tranquilizer etcAhn. J. H. et. al. (35) have reported the synthesis and DP-IV inhibition of cyno pyrazoline derivatives as potent antidiabetic agents.

 

Thus interesting biological activities of a novel heterocycles like pyrazolepyrazolines have stimulated considerable research work in recent years leading to the synthetic utility of the derivatives of these ring system. In our research for new potential antimicrobial compounds, the reaction series of chalcones with hydrazine hydrate /phenylhydrazine under different conditions has been investigated and the pharmacological interest of the compound have been studied.

 

RESULTS AND DISCUSSION:

With the biodynamic activities of chalcones and it is a good synthon for various heterocyclic rings, the interest has been focused on the synthesis of newer chalcones. With a view to obtaining compounds having better therapeutic activity.

 

The structure of all compounds (a-l) were determined using IR, PMR together with elemental analysis. IR spectra of compounds (a-l ) showed absorption bands around 1603 cm-1 regions resulting from C=N function and 1663 cm-1 regions resulting from C=O function of pyrazoline. PMR spectra of compounds (a-l) showed singlet protpn of 3H at chemical shift around 2.345 ppm for –COCH3 proton (36).

 

New Pyrazolines have been synthesized by the reaction of 1-aryl-3-(1’,N-phenyl-3’-o-methoxyphenyl-pyrazole-4’-yl)-2-propen-1-one with various aryl groups in 52 to 77%  yield. pyrazolines having high melting points. The structure of compounds are confirmed by IR, NMR and Mass spectral data and are further supported by correct elemental analysis. Newly synthesized compounds of pyrazolines (a, b, c, d, e, f, g, h, i, j, k, l). compound (a to l) have been tested their antibacterial activity against gram positive and gram negative bacteria. The compound b, f, h, i and j were shown significant activities and compound a, c, d, e, g, k and l have shown moderate activity. The same compound were tested for their antifungal activity against A. niger, and C. albicans. The compound a, e, g, i and j were shown significant activities and compound b, c, d, f, h, k and l have shown moderate activity

 

MATERIAL AND METHODS:

Melting Points were determined on Gallen-Kamp melting point apparatus and are uncorrected. All the compounds were routinely checked for their homogeneity by TLC on silica gel-G plates. IR  spectra were recorded in KBr on a Perkin-Elmer BX series FT-IR spectrophotometer, 1H NMR spectra were recorded BRUKER Spectrometer on a 400 MHz in CDCl3 using TMS as internal standard and satisfactory C,H,N and S analyses were obtained for all the compounds. The mass spectra were recorded on (FAB mass), spectromatery used to confirm their structure.

 

Antibacterial and antifungal activity (antimicrobial activity) were carried out by cup – plate agar diffusion method. The bacterial strains studied are identified strains and were obtained from National chemical laboratory (NCL), pune, India.

 

Experimental

1,N-acetyl-3-aryl-5-(1’,N-phenyl-3’-o-methoxyphenyl-pyrazole-4’-yl)-pyrazoline

Synthesis of o-Methoxyphenylhydrazone [1]:

 

A mixture of Phenylhydrazine (1.08 g, 0.01 mole) and o-Methoxyacetophenone (1.64 g, 0.01 mole) in absolute ethanol was refluxed in water bath for 2 hrs. in the presence of 1ml of glacial acetic acid. Product obtained after cooling was crystallized from absolute ethanol. Yield 92%, m.p. 420C (C15H16N2O; Calculated : C, 75.56; H, 7.13, N, 11.01%; Found : C, 75.51; H, 7.09; N,10.95%).

 

Synthesis of 1,N-Phenyl-3-o-MethoxyPhenyl-4-Formyl Pyrazole [2]:

o-Methoxyphenylhydrazone (2.54 g, 0.01 mole) was added into a Vilsmeier-Haack reagent (prepared by drop wise addition of 3 ml POCl3 in ice cooled 25 ml DMF) and refluxed for 6 hrs. The reaction mixture was poured on to crushed ice followed by neutralization using sodium bicarbonate. Crude product was filtrated and crystallised from methanol. Yield 87%, m.p. 1240C (C17H14N2O2; Calculated: C, 73.95; H, 5.52; N, 9.58;  Found : C, 73.89; H, 5.48; N, 9.52%).

 

Synthesis of 1-aryl-3-(1',N-phenyl-3'-o-methoxyphenyl-pyrazol-4'-yl)-2-propen-1-one [3]:

To a solution of 1,N-Phenyl-3-o-Methoxyphenyl-4-Formyl Pyrazole (2.92 g, 0.01 mole), p-Bromoacetophenone (1.99 g, 0.01 mole) in ethanol (25 ml) and 40% NaOH was added till the solution becomes basic. The reaction mixture was stirred for 24 hrs. The contents were poured on to crushed ice. Upon neutralization, the solid was separated and crystallized from ethanol. Yield 65%, m.p.1600C (C25H19BrN2O2; Calculated : C, 65.97; H, 4.47; N, 5.92;  Found : C, 65.91; H, 4.41; N, 5.87%).

 

Synthesis of 1,N-acetyl-3-aryl-5-(1’,N-phenyl-3’-o-methoxyphenyl-pyrazole-4’-yl)-pyrazoline [4]:

A mixture of 1-(p-Bromophenyl)-3-(1',N-Phenyl-3'-o-Methoxyphenyl-Pyrazol-4'-yl)-2-Propen-1-one (4.73 g, 0.01 mole) in 25ml absolute alcohole, hydrazine hydrate (1 g,0.02 mole) and glacial acetic acid (10 ml) was refluxed for 8 hrs.  The reaction mixture was poured on to crushed ice and product was isolated and crystallized from Ethanol. Yield 46%, m.p. 89oC (C27H23BrN4O2; Calculated : C, 63.52; H, 4.76; N, 10 .58;  Found : C, 63.46; H, 4.71; N, 10.54%). IR[KBr] Vmax Cm-1 : 2925, 2853, 1459, 1394 [C-Hstretching(asym.), C-H stretching(sym.), C-H i p.def.(asym.), C-H o.o.p.def. (sym.), -CH3 alkane], 3057, 1557, 1123, 825 [C=C stretching, C-H i p.def., C-H o.o.p.def, aromatic], 1597, 1123 [C=N stretching, C-N stretching],1245,1039, [C-O-C str. (asym.),C-O-C str.(sym.) ether], 1663,1603 [C=O stretching,C=N stretching, Pyrazoline], 753[C-Cl stretching,Halide]  PMR Spectra : [δ CDCl3], 2.34[3H s, -COCH3], 3.07-3.13[1H d, CH], 3.39-3.47[1H d, CH], 3.95[3H s, -OCH3],5.75-5.79[1H d, CH], 6.83-6.85[1H  d, Ar-H], 6.98-7.01[1H t, Ar-H],7.22-7.31[2H m, Ar-H], 7.38-7.45[5H m, Ar-H], 7.48-7.50[2H d, Ar-H], 7.68-7.70 [2H d,Ar-H],7.82[1H s,CH Similarly other substituted Pyrazolines have been prepared. The physical data are recorded in table-I.

 

Biological Evaluation

Anti-bacterial activity of 1,N-acetyl-3-aryl-5-(1’,N-phenyl-3’-o-methoxyphenyl-pyrazole-4’-yl)-pyrazoline:

Newly synthesized  compound (a to l) have been tested their antibacterial activity against gram positive and gram negative bacteria B. coccus, S. aureus, E. aerogenes, P. aeruginosa by  the help of borer in agar medium and filled with 0.04ml (40µg) solution of sample in DMF and Amoxicillin, Benzoylpenicillin , Ciprofloxacin, Erythromycin, Greseofulvin were used as a reference compound. The compound b, f, h, i and j were shown significant activities and compound a, c, d, e, g, k and l have shown moderate activity.

 

The plates were incubated at 37o C for 24 hours and the control was also maintained with 0.04 ml of DMF in a similar manner and the zones of inhibition of the bacterial

growth were measured in millimeter and are recorded data in table-II.

 

Anti Fungal activity:

The same compound were tested for their antifungal activity against A. niger, and C. albicans. The compound a, e, g, i and j were shown significant activities and compound b, c, d, f, h, k and l have shown moderate active

 

 

 


Table – I : PHYSICAL CONSTANTS OF 1,N-ACETYL-3-aryl-5-(1’,N-PHENYL-3’-o-METHOXYPHENYL-  PYRAZOLE-4’-YL)-PYRAZOLINE

Sr.

No

R

Molecular

Formula

Molecular

Weight

M.P.

°C

Rf*

Value

Yield

%

% of C, H, N, S Cal / Found

C

H

N

S

a

C6H5-

C27H24N4O2

436

186

0.73

57

79.08

5.59

11.56

-

79.01

5.53

11.52

-

b

4-NH2-C6H4-

C27H25N5O2

451

112

0.68

52

76.73

5.61

14.02

-

76.01

5.56

13.95

-

c

4-Cl-C6H4-

C27H23ClN4O2

470.5

78

0.64

70

73.80

4.89

10.79

-

73.74

4.83

10.73

-

d

4-Br-C6H4-

C27H23Br N4O2

515

89

0.57

46

67.79

4.49

9.94

-

67.75

4.42

9.89

-

e

4-F-C6H4-

C27H23FN4O2

454

106

0.60

64

76.17

5.16

11.15

-

76.11

5.12

11.12

-

f

2-OH-C6H4-

C27H24N4O3

452

158

0.49

53

76.59

5.39

11.19

-

76.54

5.34

11.14

-

g

4-OH-C6H4-

C27H24N4O3

452

132

0.55

61

76.51

5.29

11.19

-

76.43

5.24

11.12

-

h

4-OCH3-C6H4-

C28H26N4O3

466

97

0.47

76

76.86

5.64

10.89

-

76.80

5.57

10.83

-

i

4-CH3-C6H4-

C28H26N4O3

450

123

0.53

51

79.27

5.83

11.24

-

79.22

5.77

11.20

-

j

3-NO2-C6H4-

C27H23N5O4

481

137

0.59

74

72.17.

4.89

13.22

-

72.11

4.78

13.18

-

k

4-NO2-C6H4-

C27H23N5O4

481

156

0.54

77

72.27

4.82

13.22

-

72.23

4.78

13.15

-

l

2-C4H3S-

C25H22N4O2S

442

103

0.61

65

73.03

5.09

11.42

7.20

72.96

5.04

11.38

7.23

 

Tabel - II:- Antibacterial and Antifungal activity.

Compounds

Antibacterial and Antifungal activity MIC in μg/ml

B. coccus

P. aeruginosa

S. aureus

E. aerogenes

A. niger

C. albicans

a

18

16

16

14

18

19

b

21

20

18

18

16

14

c

15

14

14

15

15

13

d

16

17

17

14

14

16

e

15

17

15

18

19

18

f

18

19

19

20

15

14

g

18

19

16

16

20

19

h

19

21

21

18

17

15

i

20

21

18

19

18

19

j

17

18

19

19

19

21

k

16

14

16

17

15

14

l

18

17

16

12

15

17

Amoxicillin

25

22

25

20

00

00

Benzoylpenicillin

18

21

19

21

00

00

Erythromycin

22

23

21

19

00

00

Ciprofloxacin

20

16

15

22

00

00

Greseofulvin

00

00

00

00

26

27

 

 


Reaction Scheme of 1,N-acetyl-3-aryl-5-(1’,N-phenyl-3’-o-methoxyphenyl-pyrazole 4’-yl)-pyrazoline

 


 


CONCLUSION:

Newly synthesized compounds of pyrazolines (a to l)  have been tested for their anti bacterial activity against gram positive bacteria B. coccus, S. aureus, E. aerogenes and gram negative bacteria P. aeruginosa by the help of borer in agar medium and filled with 0.04ml (40µg) solution of sample in DMF. Amoxicillin, Benzoylpenicillin, Ciprofloxacin, Erythromycin, Greseofulvin were used as a reference compound. The compound . The compound b, f, h, i and j were shown significant activities and compound a, c, d, e, g, k and l have shown moderate activity. The compounds (a to l)  were tested for their anti fungal activity against A. niger and C. albicans using cup-plate method. The compound a, e, g, i and j were shown significant activities and compound b, c, d, f, h, k and l have shown moderate activity.  All the other compounds did not show significant activity against the fungi at the concentration used.

ACKNOWLEDGEMENT:

The authors are thankful to Director, C.D.R.I., Lucknow (U.P.) and National chemical laboratory (NCL), pune, for providing spectral and analytical data of the compounds. They are also thankful to the microcare laboratory, Surat (Gujarat) for the biological active

 

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Received on 09.02.2011         Modified on 05.03.2011

Accepted on 19.03.2011         © AJRC All right reserved

Asian J. Research Chem. 5(5): May 2012; Page 571-575