E-ISSN: 2278-3229

International Journal of Green and Herbal Chemistry

Available online at www.ijghc.org Green Chemistry
IJGHC; June-August, 2012; Vol.1.No.2, 108-119 Research Article

Microwave Assisted Synthesis of Novel Benzotriazole - Pyrazoline derivatives and its Pharmacological Evaluation
Kalmendra Singh Sisodiya1*, Manisha2, Amit Sahu3, V.K.Sharma4 and Bharat Parashar5
Department of Chemistry, Pacific University, Udaipur, India. Skyline Institute of Engineering and Technology, Greater Noida (U.P.), India 3 Department of Chemistry, Techno India NJR instituted of technology Udaipur, India 4 Department of Chemistry, M.L. Sukhadia University, Udaipur, (Rajasthan) 5 Department of pharmaceutics, M.B.University; Solan (H.P.).
2 1

Received: 13 March 2012; Revised: 31 March 2012; Accepted: 8 April 2012

Abstract: A solvent free approach for organic synthesis is described here which involve microwave exposure of neat reactants. A novel and simple method have been developed for the synthesis of some Benzotriazole - Pyrazoline derivatives under microwave irradiation. In addition, these compounds were obtained with conventional heating procedures and microwave irradiation to compare them with those obtained with solid support synthesis. All the compounds synthesized were characterized by running TLC, Elemental analysis, IR, NMR and MS spectra. Consequently, the solid support synthesis method provided nearly the same and higher product yields in a very short period of time. These results suggest that in addition, the use of solid supports in conjunction with microwave leads to a higher yield, remarkable reactions rate enhancement and high catalytic activity with the optimum utilization of energy. Solid support synthesis method is more useful than the microwave irradiation method and conventional method. Keywords: Green chemistry, microwave irradiation, solid supported synthesis,
benzotriazole - pyrazoline derivatives, neat reaction technology

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INTRODUCTION
Classical methods require elaborate apparatus setup, longer heating time and large volume of organic solvents and there is virtually no control over the energy input. Hazardous chemicals and by products of various reactions increasing pollution in the environment. Keeping in view the need for avoiding hazardous chemicals and solvents in chemical reactions, microwave technique was found to be accelerating a wide variety of transformations. Neat reaction technology is a step forward in the direction of solvent free reactions and an alternative approach that eliminates the use of a solid support as well as solvent from the reaction. In recent years design of environmentally benign reactions is an important goal in organic synthesis. Microwave induced Organic Reaction Enhancement (MORE) chemistry offers a simple, non-conventional technique for the synthesis of a wide variety of compounds having medicinal, pharmaceutical and commercial importance. In this expeditious and solvent free approach the reactants were adsorbed over inorganic supports and exposed to microwave irradiation. In recent years the use of microwaves1,2 has been well established as a pollution free technique which allows reaction to occur on a preparative scale in open vessels under solvent free conditions which avoids the risk of high pressures and explosions. Microwave activation rather than conventional heating is preferred, as solid supports are rather poor thermal conductors but strong microwave absorbents, which results in lesser evaporation of solvents preventing pollution3. Further the reactions are generally faster and the products obtained are of high purity4, 5. Pyrazole as well as pyrazolines containing compounds have been reported to show a broad spectrum of biological activities such as antimicrobial6-9, anti-tumor10, and antiinflammatory11, 12 agents. Due to bioactivity associated with pyrazole and pyrazolines containing compounds, researchers and chemist are very much interested in pyrazole chemistry13-15. Pyrazolone are associated with broad spectrum of biological activities16, Pyrazolone exhibit anti-inflammatory17 and analagics activity 18, anticancer activity19, 20 antiamoebic activity21 , antitubercular activity22 and antitumor activity23 . Synthesis of some Benzotriazole - Pyrazoline derivatives under microwave irradiation using diethyl malonate, acetyl acetone, 1- Chloro-hexane 2-4 dione, ethyl 2-cyanoacetate and phenyl isothiocynate by solid support synthesis method, conventional heating procedures and microwave irradiation method were done. The reaction carrierd out in absolute alcohol or DMF using conventional method required 4-10 h, while microwave irradiation method require only 3.30-7 min and solid support method using silica gel require only 4-6.50 min. The synthetic route of above mentioned compound is shown in Scheme 1, (Table 1). MATERIALS AND METHODS Experimental Section: All the irradiation was carried out in a modified microwave oven (Kenstar, model no: OM26.EGO). Melting points of synthesis compounds were determined in open capillaries in liquid paraffin and are uncorrected. Purity of the compounds in addition to elemental analysis were verified by percolated TLC using silica gel G as a adsorbent using ethyl acetate: n-hexane (7:3) as a eluent and spot was detected by using iodine vapours. The IR (KBr pellets) spectra were recorded on a Perkin Elimer1800- spectrophotometer and H1NMR spectra were recorded on BRUKER DRX-300MHz spectrophotometer, (TMS as a internal reference) and chemical shifts are expressed in . Mass spectra were recorded on Jeol D30 spectrophotometer. Elemental analyses for C, H and N were conducted using a Perkin -Elmer C, H, and N analyzer. Their result was found to be an in good agreement with the calculated values (0.4%).

Synthesis of ethyl 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetate (I)

Conventional Method: A mixture of Benzotriazole (0.01 mol), ethyl chloro acetate ( 0.01 mol) and potassium carbonate 3.0 gm in acetone 50.0 mL was stirred for 6 hours. Completion of reaction was 5 109 IJGHC; 2012, Vol.1, No.2, 108-119.

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Synthesis of Compound (I) and (II)

H2N
+

NH O

O O Cl

K2CO3

N N

OO
(I)

NH2NH2.H2O

N N
(II)

HN N

Scheme No. 1
O N HN N O O

N N

O N N N O

N N

1-(2-(1Hbenzo[d][1,2,3]triazol-1O yl)acetyl)pyrazolidine-3,5dione
(1)

N S N NH N O O N N

1-(2-(1H-benzo[d][1,2,3]triazol-1yl)acetyl)-2-acetylpyrazolidine-3,5dione
(2)

CH2 (COOC2H5 )2

(E)-N-(4-oxo-2-(phenylimino)thiazolidin-3yl)-1H-benzo[d][1,2,3]triazole-1carboxamide
(8) (i)Morpholine

H3C N N N N N O CH3
CH 3 CO

2-(1H-benzo[d][1,2,3]triazol-1yl)-1-(3,5-dimethyl-4,5dihydropyrazol-1-yl)ethanone
(3)

CH 2 CO C

O O N N Cl N N N

H3 CH 2C H CO COC 2 ClCH 2

O 2-(1HCN 1-(2-(1H-benzo[d][1,2,3]triazol-1benzo[d][1,2,3]triazol-1- CH2 C yl)acetyl)-4-phenylthiosemicarbazide O OC yl)acetohydrazide H

2

)2 H5 C2 OO (C 2 CH

OH CO CH 3

N
H
3

(ii)ClCH2COOC2H 5

N H2N N
(II)

C6H5NCS

N H N HN N H S

NH O

CH

(7)

CH3COCH2COCH2CH3

O N N N N N

1-(2-(1H-benzo[d][1,2,3]triazol1-yl)acetyl)-3-(chloromethyl)1H-pyrazol-5(4H)-one
(4)

H3C N H2N N N O N N O

1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)3-amino-1H-pyrazol-5(4H)-one
(6)

1-(2-(1H-benzo[d][1,2,3]triazol-1yl)acetyl)-3-methyl-1H-pyrazol-5(4H)one
(5)

Scheme-2

The synthetic route of compounds (1-8) 110 IJGHC; 2012, Vol.1, No.2, 108-120.

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Table: 1- Comparative Analysis of Conventional and Microwave Methods Comp Molecular formula M.wt Condition Conventional method Microwave method m. p o C On Solid Support Yield R time (%) (min) 88 6.10 86 90 88 90 92 90 86 6.30 3.30 5.30 5.20 2.20 2.10 3.10 Rf

In solvent Yield (%) 70 68 71 72 71 73 69 R time(h) 6.00 5.10 8.10 5.22 6.12 4.55 4.11 5.50 Yield (%) 82 80 84 81 83 84 82 80 R time (min) 3.00 5.10 3.10 3.30 3.00 3.00 2.00 2.30

1 2 3 4 5 6 7 8

C11H9N5O3 C13H11N5O4 C13H13N5O C12H10ClN5O2 C12H11N5O2 C11H10N6O2 C15H14N6OS C16H12N6O2S

259 301 257 291 257 258 326 352

A.A A.A A.A -

Morpholin 70 e A.A= Acetic acid

17 2 18 9 16 9 20 1 16 9 18 2 21 0 23 2

0.64 0.62 0.68 0.61 0.68 0.66 0.66 0.66

Synthesis of 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl) acetyl)pyrazolidine-3,5-dione. (1)

Conventional Method: A mixture of (II) (0.01 mol), diethyl malonate (0.01mol) and two to three drops of glacial acetic acid were taken in a round bottom flask in 50 mL of ethyl alcohol and was refluxed for about 6 hr on a water bath. After completion of the reaction as indicated by TLC, the reaction mixture was cooled and treated with ice cold water. The brown colour oily mass obtained was cooled and the crud product was recrystallized from ethanol to give afford 1. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: A mixture of (II) (0.01 mol), diethyl malonate (0.01mol) and two to three drops of glacial acetic acid in 30 mL of ethyl alcohol was subjected to microwave irradiation at 360 W for 3 min with an intermittently at 30 sec intervals. After completion of the reaction as indicated by TLC, the reaction mixture was cooled and treated with cold water. The brown colour oily mass obtained was cooled and the crud product was recrystallized from ethanol to give afford 1. Physical characteristic of compound is recorded in Table No 1. Solid Support Method: A mixture of (II) (0.01 mol), diethyl malonate (0.01mol) were dissolve in alcohol and the mixture was absorbed on acidic alumina and was subjected to microwave irradiation at 500 W for 5.10 min with an intermittently at 10 sec intervals. After completion of the reaction as indicated by TLC, the compound was separate out from di ethylethane and recrystallise from ethyl alcohol. Physical characteristic of compound is recorded in Table No 1. Physical and Elemental Analysis of Compound 1: Mol. Wt. 259; M.P. 172-174 C; Yield: 88 %; Rf 3035 (Aromatic CH-Ar. str. ), 2260 (N=N), 1735, 1730, :0.64; IR ( KBr cm-1): 3303 (-NH), 1 1666,(C=O), ; H NMR (400 MHz, DMSO-d6) : 7.45- 7.80 (4H, m, Ar-H); 8.70 (s, H, NH), 4.75 (s, 111 IJGHC; 2012, Vol.1, No.2, 108-119.

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2H, CH2 ) , 3.35 (s, 2H, CH2), MS (m/z+) [M+] 259; Anal.Calc.for C11H9N5O3; C, 50.97; H, 3.50; N, 27.02; Found: C, 51.26.; H, 3.30; N, 26.82.

Synthesis of 1- (2- (1H-benzo [d][1,2,3] triazol-1-yl)acetyl) -2-acetylpyrazolidine -3,5-dione (2)

Conventional Method: A mixture of (II) (0.01 mol) and diethyl malonate (0.01 mol) in glacial acetic acid (5 mL) were taken in a round bottom flask and was refluxed for about 5 hr on a water bath . After completion of the reaction as indicated by TLC, the reaction mixture was cooled and treated with cold water. The solid that precipitated was filtered, washed with water and recrystallized from ethanol to afford 2. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: A mixture of (II) (0.01 mol) and diethyl malonate (0.01 mol) in glacial acetic acid (5 mL) was subjected to microwave irradiation at 360 W for 5.10 min with an intermittently at 20 sec intervals. After completion of the reaction as indicated by TLC, the reaction mixture was cooled and treated with cold water. The solid that precipitated was filtered, washed with water and recrystallized from ethanol to afford 2. Physical characteristic of compound is recorded in Table No 1. Microwave Method on Solid Support: Compound (II) (0.01 mol) and diethyl malonate (0.01 mol) were dissolved in acetone and the solution was absorbed on acidic alumina and was subjected to microwave irradiation at 500 W for 4.30 min with an intermittently at 10 sec intervals . After completion of the reaction as indicated by TLC compound was separate out from acetone and allows drying. Product was recrystallised with di methyl ether to afford 2. Physical characteristic of compound is recorded in Table No 1. Physical and Spectral Analysis of Compound 2: Mol. Wt. 301; M.P. 189-191 C; Yield: 86 %; Rf :0.62; IR ( KBr cm-1): 3130 (Aromatic CH-Ar, str. ), 2270 (N=N), 1740,1693, 1670, 1640, (C=O), ; 1H NMR (400 MHz, DMSO-d6) : 7.32- 7.82 (4H, m, Ar-H); 4.60 (s, 2H, CH2 ) , 3.30 (s, 2H, CH2), 2.40 (s, 3H, CH3) : MS (m/z+) [M+] 301; Anal. Calc.for C13H11N5O4; C, 51.83; H, 3.68; N, 23.25; Found: C, 52.06.; H, 4.02; N, 23.72;

Synthesis of 2- (1H-benzo [d][1,2,3 ]triazol-1-yl) -1- (3,5-dimethy l-4,5-dihydropyrazol-1-yl) ethanone(3)

Conventional Method: A mixture of 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (II) (0.01 mol) and acetyl acetone (0.01 mol) was taken in Erlenmeyer flask. The mixture was well stirred and then taken in a round bottom flask and was refluxed for about 2 hr on a water bath. After completion of the reaction as indicated by TLC the reaction- mixture was then allowed to stand at RT and the product formed was washed with cold water and dissolved in methanol then filtered, dried and recrystallized from ethanol to afford 3. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: A mixture of 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (II) (0.01 mol) and acetyl acetone (0.01 mol) was taken in Erlenmeyer flask. The mixture was well stirred subjected to microwave irradiation at 360 W for 3.10 min with an intermittently at 10 sec intervals. After completion of the reaction as indicated by TLC the reaction- mixture was then allowed to stand at RT and the product formed was washed with cold water and dissolved in methanol then filtered, dried and recrystallized from ethanol to afford 3. Physical characteristic of compound is recorded in Table No 1. Microwave Method on Solid Support: A solution of (II) (0.01 mol) and acetyl acetone (0.01 mol) were taken in beaker. The mixture was well stirred and then absorbed on neutral alumina and was subjected to microwave irradiation at 500 W for 3.10 min with an intermittently at 20-25 sec intervals. After completion of the reaction as indicated by TLC the compound was separate out from acetone and recrystallized from ethanol to afford 3. Physical characteristic of compound is recorded in Table No 1. 112 IJGHC; 2012, Vol.1, No.2, 108-120.

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Physical and Spectral Analysis of Compound 3: Mol. Wt. 257; M.P. 169-171 C; Yield: 90 %; Rf :0.68; IR ( KBr cm-1): 3110 (Aromatic CH-Ar, str. ), 2265 (N=N), 1690, (C=O), 1553(C=N), : 1H NMR (400 MHz, DMSO-d6) : 7.32- 7.82 (4H, m, Ar-H); 5.71(s, 1H, CH) , 4.56 (s, 2H, CH2 ) , 3.42 (s, 2H, CH2), 3.35 (s, 3H, CH3), 2.40 (s, 3H, CH3 ): MS (m/z+) [M+] 257 ; Anal. Calc.for C13H13N5O; C, 60.69; H, 5.88; N, 27.22; Found: C, 60.92; H, 5.62; N, 27.50.

Synthesis of 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-3-(chloromethyl)-1H-pyrazol5(4H)-one (4)

Conventional Method: A mixture of (II) (0.01 mol), 1- Chloro-hexane 2-4 dione (0.012 mol) was taken in a round bottom flask and was refluxed for about 5 hr on a water bath. TLC monitored the reaction. After completion of the reaction, reaction- mixture was allowed to stand at RT and the product formed was filtered, washed with water, dried and recrystallized from ethanol to afford 4. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: A mixture of (II) (0.01 mol), 1- Chloro-hexane 2-4 dione (0.07 mol) was taken in a borosil beaker (100 mL). The well-stirred mixture was irradiated inside the microwave oven for a period of 3.30 minutes (at 480 W). TLC monitored the reaction. After completion of the reaction, reaction- mixture was allowed to stand at RT and the product formed was filtered, washed with water, dried and recrystallized from ethanol to afford 4. Physical characteristic of compound is recorded in Table No 1. Microwave Method on Solid Support: Compound (II) (0.01 mol), 1- Chloro-hexane 2-4 dione (0.07 mol) were dissolved in alcohol and the mixture was absorbed on neutral alumina and was irradiated inside the microwave oven for a period of 5.30 minutes (at 500W). TLC monitored the reaction. After completion of the reaction, product was separate out from aceton recrystallized from ethanol to afford 4. Physical characteristic of compound is recorded in Table No 1. Physical and Spectral Analysis of Compound 4: Mol. Wt. 291; M.P. 201-122 C; Yield: 88 %; Rf :0.61; IR ( KBr cm-1): 3080 (Aromatic CH-Ar str. ), 2240 (N=N), 1695,1666 (C=O), 1606 (C=N), : 1H NMR (400 MHz, DMSO-d6) : 7.7-7.8(m, 4H,), 2.80 (s, 2H, CH2), 3.45 (s, 2H, CH2): 7.32- 7.82 (4H, m, Ar-H); 5.71(s, 1H, CH) , 4.30 (s, 2H, CH2 ) , 3.22 (s, 2H, CH2), : MS (m/z+) [M+] 291 ; Anal. Calc.for C12H10 ClN5O2; C, 49.41; H, 3.46; N, 24.01; Found: C, 49.64; H, 3.16; 1-; N, 24.24.

Synthesis of 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-3-methyl-1H-pyrazol--(4H)-one (5)

Conventional Method: Compound (II), hexane-2, 4-dione (0.01 mol), and (0.005 mol) acetic acid was mixed thoroughly. This mixture was taken in a round bottom flask and was refluxed for about 6 hr on a water bath. Upon completion of the reaction (monitored by TLC), the reaction mixture was poured onto crushed ice. The solid mass obtained was filtered and washed several times with water. Purification by recrystallization with alcohol gave product 5. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: Compound (II), hexane-2, 4-dione (0.01 mol), and (0.00- mol) acetic acid was mixed thoroughly. This mixture was subjected to microwave irradiation (by keeping inside a microwave oven) for 3.0 min at 480 W power levels with intermitted irradiation for 30 second. Upon completion of the reaction (monitored by TLC), the reaction mixture was poured onto crushed ice. The solid mass obtained was filtered and washed several times with water. Purification by recrystallization with alcohol gave product 5. Physical characteristic of compound is recorded in Table No 1. Microwave Method on Solid Support: Compound (II), hexane-2,4-dione (0.01 mol), were dissolved in acetone and absorbed on acidic alumina and irradiated inside the microwave oven for a period of 5.20 min at 500 W power levels with intermitted irradiation for 30 second. Upon completion of the reaction (monitored by TLC), the reaction mixture was poured onto crushed ice. The solid mass obtained was 113 IJGHC; 2012, Vol.1, No.2, 108-119.

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filtered and washed several times with water. Purification by recrystallization with alcohol gave product 5. Physical characteristic of compound is recorded in Table No 1. Physical and Spectral Analysis of Compound 5: Mol. Wt. 257; M.P. 169-171 C; Yield: 90 %; Rf :0.68; IR ( KBr cm-1): 3115 (Aromatic CH-Ar str. ), 2262 (N=N), 1680, 1606 (C=O), 1530 (C=N), ; 1H NMR (400 MHz, DMSO-d6) : 7.40- 7.88 (4H, m, Ar-H); 4.56 (s, 2H, CH2 ) , 3.42 (s, 2H, CH2), 3.35 (s, 3H, CH3), : MS (m/z+) [M+] 257 ; Anal. Calc.for C12H11N5O2; C, 56.03; H, 4.31; N, 27.22; Found: C, 56.36; H, 4.64; N, 27.46.

Synthesis of 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-3-amino-1H-pyrazol--(4H)-one (6)

Conventional Method: 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (0.01 mol) (II)and (0.01) and ethyl 2-cyanoacetate (0.01) in -0 mL ethanol were taken in a round bottom flask and was refluxed for about 4hr on a water bath and reaction progress was monitored by TLC. Then crude product so obtained was poured into ice-cold water, filtered, dried, and recrystallized from methanol to give product 6. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (0.01 mol) (II) and (0.01) and ethyl 2-cyanoacetate (0.01) were mixed together to form a paste and then subjected to microwave irradiation for 3.0 min at 400 W power level and reaction progress was monitored by TLC. Then crude product so obtained was poured into ice-cold water, filtered, dried, and recrystallized from methanol to give product 6. Physical characteristic of compound is recorded in Table No 1. Solid Support Method: 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (0.01 mol) (II) and (0.01) and ethyl 2-cyanoacetate (0.01) were dissolved in alcohol and absorbed on alumina. The mixture was subjected to microwave irradiation at 500 W for 4.20 min with an intermittently at 30-35 sec intervals. After completion of the reaction indicated by TLC the compound was separate out from acetone and recrystallize from alcohol to give product 6. Physical characteristic of compound is recorded in Table No 1. Physical and Spectral Analysis of Compound 6: Mol. Wt. 258; M.P. 182-183 C; Yield: 92 %; Rf :0.66; IR ( KBr cm-1): 3334, 3218(-NH2), 3140 (Aromatic CH-Ar str. ), 2262 (N=N), 1698, 1672 (C=O), 1554 (C=N), ; 1H NMR (400 MHz, DMSO-d6) : 7.46- 7.68 (4H, m, Ar-H); 5.12 (s, 2H, NH2), 4.50 (s, 2H, CH2 ) , 3.42 (s, 2H, CH2),: MS (m/z+) [M+] 258 ; Anal. Calc.for C11H10N6O2; C, 51.16; H, 3.90; N, 32.54; Found: C, 51.36; H, 4.20; N, 32.74.

Synthesis of 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-4-phenylthiosemicarbazide (7)

Conventional Method: To a mixture of 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (II) (0.01 mol ) and phenyl isothiocynate (0.01 mol e) in a 40 ml DMF was were taken in a round bottom flask and was refluxed for about 4 hr on a water bath. TLC examined the progress of the reaction. The mixture was poured into ice-cold water. The resulting solid was filtered, dried and recrystallized from benzene to give product 7. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: To a mixture of 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (II) (0.01 mol ) and phenyl isothiocynate (0.01 mol e) in a 20 ml DMF was taken in Erlenmeyer flask. The mixture was irradiated under microwave for 2.00 min at 360 W powers with intermittent radiation of 15 sec interval. TLC examined the progress of the reaction. The mixture was poured into ice-cold water. The resulting solid was filtered, dried and recrystallized from benzene to give product 7. Physical characteristic of compound is recorded in Table No 1. Microwave Method on Solid Support: 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetohydrazide (II) (0.01 mol) and phenyl isothiocynate (0.01 mol) were dissolved in acetone and the mixture was absorbed on neutral alumina and was subjected to microwave irradiation at 500 W for 5.10 min with an intermittently at 15-20 sec intervals. After completion of the reaction the compound was separate out from acetone and 114 IJGHC; 2012, Vol.1, No.2, 108-120.

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recrystallize from alcohol to give product 7. Physical characteristic of compound is recorded in Table No 1. Physical and Spectral Analysis of Compound 7: Mol. Wt. 326; M.P. 210-2111 C; Yield: 90 %; Rf :0.66; IR ( KBr cm-1): 3416,3372, 3312(-NH), 3089 (Aromatic CH-Ar str. ), 2262 (N=N), 1708 (C=O), 1100 (C=S), ; 1H NMR (400 MHz, DMSO-d6) : 10.20 (s,H, HN-CO), 7.48- 7.80 (m, 9H, Ar-H); 4.34 (s,H,HN-Ph), 3.42 (s, 2H, CH2), 2.20 (s,H,HN-N); : MS (m/z+) [M+] 326 ; Anal. Calc.for; C15H14N6OS ; C, 55.20; H, 4.32; N, 25.75; Found: C, 55.20; H, 4.32; N, 25.75.

Synthesis of (E) -N- (4-oxo-2-(phenylimino) thiazolidin-3-yl)-1H-benzo[d][1,2,3]triazole-1carboxamide (8)

Conventional Method: A mixture of 7 (0.01 mol) and ethylchloroacetate in presence of catalytic amount of morpholine in 40 mL of ethanol were taken in a round bottom flask and was refluxed for about 5.50 hr on a water bath. After completion of the reaction as indicated by TLC, the reaction mixture was cooled and treated with ice-cold water and recrystallized from ethanol to give product 8. Physical characteristic of compound is recorded in Table No 1. Microwave Method in solvent: A mixture of 7 (0.01 mol) and ethylchloroacetate in presence of catalytic amount of morpholine were mixed thoroughly. Than the mixture was irradiated under microwave oven for 2.30 min at 480 W with an intermitted irradiation for 30sec. interval. After completion of the reaction as indicated by TLC, the reaction mixture was cooled and treated with ice-cold water and recrystallized from ethanol to give product 8. Physical characteristic of compound is recorded in Table No 1. Microwave Method on Solid Support: A mixture of 7 (0.01 mol) and ethylchloroacetate were mixed thoroughly and dissolved in alcohol and the mixture was absorbed on basic alumina and subjected to microwave irradiation at500 W for 4.10 min with an intermittently at 15-20 sec intervals. After completion of the reaction as indicated by TLC the compound was separate out from acetone and 1,2 dichloro methane. Physical characteristic of compound is recorded in Table No 1. Physical and Spectral Analysis of Compound 8: Mol. Wt. 352; M.P. 232-334 C; Yield: 86 %; Rf :0.66; IR ( KBr cm-1): 3080 (Aromatic CH-Ar str. ), 2282 (N=N), 1698, 1649 (C=O), 740 (S-C-S), ; 1H NMR (400 MHz, DMSO-d6) : 10.20 (s, H, HN-CO), 9.67(s, H, HN) , 7.32- 7.88 (m, 9H, Ar-H); 4.10 (s, 2H, CH2), : MS (m/z+) [M+] 352 ; Anal. Calc. for; C16 H12 N6 O2 S; C, 54.54; H, 3.43; N, 23.85; Found: C, 54.74; H, 3.24; N, 23.62.

BIOLOGICAL ASSAY
All the newly synthesized compounds were tested in vitro for antimicrobial activity against four bacterial strains two-gram positive bacteria (Bacillus subtilis and staphylococcus aureus) and two-gram negative bacteria (Escherichia coli and P.aeruginosa) and antifungal activity against Candida albicans and Asperigillus niger at the concentration of 200,100,50, g/ml by using the cup plate agar disk diffusion method24,25 and determination of minimum inhibitory concentration (MIC) by broth dilution method.The concentration used in screening was choosen after determining MIC of each compound. The dimethylsulfoxide (DMSO) was used as a solvent. For this method, Mueller-Hinton agar was used as the growth medium for the bacterial strains and Sabouraud agar was growth medium for fungal species. The sterilized agar media were poured in to petridishes and allow solidifying. On the surface of the media microbial suspension were spread with the help of sterilized triangular loop. (Inoculums of standard suspension 0.1 mL of the test organism strain which contains 106 bacilli/mL) A stainless steel cylinder of 9 mm diameter (pre sterilized) was used to bore the cavities. In to these well were added 0.1mL portion of the test compounds in solvent DMSO. The drug solution was allowed to diffuse for about an hour in to the medium. The petridishes used for antibacterial screening were 115 IJGHC; 2012, Vol.1, No.2, 108-119.

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incubated at 37 1C for 24 h, while those for antifungal activity were incubated at 28 1C for 48-72 h. DMSO was used as a control for all the type of microorganisms. The control showed no activity against the strains of microorganisms used. Antimicrobial activity and antifungal activity was measured as a function of diameter of zone of inhibition (mm). The result was compared with standard drugs Ciprofloxacin for antibacterial activity and fluconazole for antifungal activity by measuring the zone of inhibition in mm at 200,100 and 50 g/mL (Table -2 and Table-3). The lowest concentration, which showed no visible growth, was taken as an end point minimum inhibitory concentration (MIC). For antibacterial activity, in present protocol 50 g/ml is considered as active as compared to the standard drug Ciprofloxacin. For antifungal activity, 50 g/mL is considered as active as compared to standard fluconazole

Table: 2. Antifungal Activities of the synthesized compounds (1-8).

Antifungal activity C. albicans 100 200 g/ml g/ml ++ +++ ++ ++++ ++ +++ ++ ++ ++ ++ +++ +++ +++ +++ +++ +++ ++++ ++++

Compounds 50g /ml + + ++ ++ ++ + ++++

1 2 3 4 5 6 7 8 Fluconazole

50g /ml ++ + ++ ++ +++ ++++

A. niger 100g /ml ++ +++ ++ ++ ++ ++ +++ ++ ++++

200g /ml ++ ++++ +++ ++ ++ +++ +++ +++ ++++

(-)< 6mm; (+) = 7 - 10 mm; (++) =11 15 mm; (+++) = 16 - 21mm; (++++) = 22 - 28mm.

RESULTS AND DISCUSSIONS

The compound ethyl 2-(1H-benzo[d][1,2,3]triazol-1-yl) acetate (I) was synthesized by microwave radiation of mixture of Benzotriazole, ethyl chloro acetate and potassium carbonate in acetone. The M.P. of the synthesized compound was checked by the given literature. The compound 2-(1Hbenzo[d][1,2,3]triazol-1-yl) acetohydrazide (II) was synthesized by ethanolic solution of compound (I) and hydrazine hydrate under microwave radiation of 240W. The purity of the compound was analyzed by TLC using ethyl acetate: n-hexane (7:3) as a mobile phase. The final compounds (1-8) were synthesized as given below. The structure of the synthesized compounds was confirmed on the basis of spectral and elemental analysis. The compound (1) 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl) acetyl)pyrazolidine-3,5-dione is synthesized by irradiating mixture of compound (II), diethyl malonate and 2-3 drops of glacial acetic acid. The IR spectrum exhibited bands due to 1735, 1730, 1666 (C=O), and 1H NMR gives shift at 4.75 (s, 2H, CH2), 3.35 (s, 2H, CH2 o) and molecular ion peak [M+] was found at 259 which confirmed the formation of compound (1). The compound (2) 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-2-acetylpyrazolidine-3,5-dione was synthesized by irradiating mixture of compound (II), diethyl malonate and 5ml of glacial acetic acid. The IR spectrum exhibited bands due to 1740,1693,1670,1640 (C=O), 1H NMR gives shift at 4.60 (s, 2H, CH2 ) , 3.30 (s, 2H, CH2), 2.40 (s, 3H, CH3) molecular ion peak [M+] was found at 301 which confirmed the formation of compound (2).

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Kalmendra Singh et al. Table: 3. Antibacterial Activities of the synthesized compounds (1-8).

Antibacterial activity Comp E. coli ounds 50 100 g g /ml /ml 1 _ ++ 2 _ ++ 3 + ++ 4 + +++ 5 _ + 6 _ ++ 7 _ + 8 _ +++ Ciprofl ++++ ++++ oxacin

P. aeruginosa 200 g /ml ++ +++ ++++ ++++ +++ +++ +++ ++++ ++++ 50 g /ml _ _ _ _ _ _ _ + ++++ 100 g /ml + _ ++ + + + + ++ ++++ 200 g /ml ++ + ++ ++ +++ + +++ +++ ++++

B. subtitis 50 g /ml _ _ _ _ _ _ _ + ++++ 100 g /ml ++ ++ ++ ++ ++ ++ ++ ++ ++++ 200 g /ml ++ +++ +++ ++++ ++ +++ +++ +++ ++++

S. aureus 50 g /ml _ _ _ _ _ _ _ _ ++++ 100 g /ml _ ++ ++ ++ ++ _ + + ++++ 200 g /ml + +++ ++ +++ +++ +++ +++ +++ ++++

(-)< 6mm; (+) = 7 - 10 mm; (++) =11 15 mm; (+++) = 16 - 21mm; (++++) = 22 - 28mm.

The compound (3) 2-(1H-benzo[d][1,2,3]triazol-1-yl)-1-(3,5-dimethyl-4,5-dihydropyrazol-1-yl)ethanone was synthesized by irradiating mixture of compound (II) and acetyl acetone. The IR spectrum exhibited bands due to 1553(C=N) ,1H NMR (400 MHz, DMSO-d6) showed peaks due to 3.35(s, 3H, CH3), 2.40 (s, 3H, CH3 ) ) and molecular ion peak [M+] was found at 257 which confirmed the formation of compound (3). The compound (4) 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-3-(chloromethyl)-1H-pyrazol-5(4H)-one was synthesized by microwave irradiation of mixture of compound (II) and 1- Chloro-hexane 2-4 dione. The IR spectrum exhibited bands due to 1695, 1666 (C=O), 1606(C=N), 1H NMR (400 MHz, DMSO-d6) showed peaks due to 4.30 (s, 2H, CH2 ) , 3.22 (s, 2H, CH2) and molecular ion peak [M+] was found at 291; which confirmed formation of compound (4). The compound (5) 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-3-methyl-1H-pyrazol-5(4H)-one was synthesized by microwave irradiation of mixture of compound (II), hexane-2, 4-dione and acetic acid. The IR spectrum exhibited bands due to 1680,1606(C=O),1530(C=N) and 1H NMR (400 MHz, DMSOd6) showed peaks due to 3.42 (s, 2H, CH2), 3.35 (s, 3H, CH3) and molecular ion peak [M+] was found at 257 which confirmed formation of compound (5). The compound (6) 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-3-amino-1H-pyrazol-5(4H)-one was synthesized by microwave irradiation of mixture of compound (II) and ethyl 2-cyanoacetate. The IR spectrum exhibited bands due to 1698, 1672 (C=O), 1554(C=N) and NMR (400 MHz, DMSO-d6) showed peaks due to 5.12 (s, 2H, NH2), 4.50 (s, 2H, CH2 ) , 3.42 (s, 2H, CH2) and molecular ion peak [M+] was found at 258 which confirmed formation of compound (6). The compound (7) 1-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetyl)-4-phenylthiosemicarbazide was synthesized by microwave irradiation of mixture of compound (II) and phenyl isothiocynate in DMF . The IR spectrum exhibited bands due to 1100(C=S), NMR (400 MHz, DMSO-d6) showed peaks due to (s, 117 IJGHC; 2012, Vol.1, No.2, 108-119.

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2H, CH2), 2.20 (s, H, HN-N), and molecular ion peak [M+] was found at 326 which confirmed formation of compound (7). The compound (8) (E)-N-(4-oxo-2-(phenylimino) thiazolidin-3-yl)-1Hbenzo[d][1,2,3]triazole-1carboxamide was synthesized by irradiating mixture of compound (7) andethylchloroacetate in presence of catalytic amount of morpholine. The IR spectrum exhibited bands due to1698, 1649(C=O), 740(S-C-S), NMR (400 MHz, DMSO-d6) showed peaks due to 4.10 (s, 2H, CH2) and molecular ion peak [M+] was found at 352 which confirmed formation of compound(8).

CONCLUSION
Most of the synthesized compounds were found to possess mild to moderate antibacterial and antifungal activity except a couple of compounds which showed excellent activity, almost equivalent to the compounds will take place which may lead to a potentially improved compounds.

ACKNOWLEDGEMENTS
The authors are thankful to Dr.Suman Jain, Director of shri ram college of Pharmacy, for providing laboratory facilities authors are also thankful to Director, CDRI Lucknow, India for providing spectral and analytical data. Special thanks to Dr. Ashok Kumar Sharma (A.D.) Regional Disease Diagnostic Center, Kota (Rajasthan) for antimicrobial screening.

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*Correspondence Author: Kalmendra Singh Sisodiya; Department of Chemistry,

Pacific University, Udaipur, India.

Email:kalmendrasisodiya@yahoo.in

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