Synthesis and Antimicrobial activity of Pyrimidine Derivatives from Amine

 

Sanjay Chauhan1*, A. K. Gyananchandran1, Pawan Tiwari1, Sanjeev Sharma2 and S.S. Shukla3

1Pranav Institute of Pharmaceutical Science and Research, Sitholi (Gwalior)

2Shri Ramnath Singh Institute of Pharmaceutical Science and Technology, Sitholi (Gwalior)   

3Columbia Institute of Pharmacy, Tekari, Raipur, Chhattisgarh

*Corresponding Author E-mail: Chauhan.sanjay84@gmail.com

 

ABSTRACT:

Pyrimidine is a heterocyclic aromatic organic compound which containing two nitrogen atoms at positions 1 and 3 of the six-member ring. In step-1, equimolar (0.02 mole) mixtures of Aromatic primary amines were refluxed in 40ml of dry ethanol for 18 hours. In step-2, an equimolar (0.02moles) mixture of Schiff’s base and Thioglycolic acid were refluxed for 18hr in 30ml of dry benzene. Then the excess benzene was distilled off under reduced pressure and the contents were poured over crushed ice and centrifuged. In step-3, Aromatic aldehydes and anhydrous sodium acetate (0.082gm) in 20ml of glacial acetic acid was refluxed for 3hrs. Then the reaction mixture was concentrated, cooled and poured into ice cold water. In step-3 and step-4, Thiourea or ureas were dissolved in ethanol (50ml). To this NaOH dissolved to minimum quantity of water was added. Then the mixture was refluxed for 6hrs and poured into 250ml of cold water. After complete these four steps, we collect the some pyrimidine derivatives compounds and show antibacterial and antimicrobial activity.

 

 


INTRODUCTION:

Pyrimidine is a heterocyclic aromatic organic compound similar to benzene and pyridine, containing two nitrogen atoms at positions 1 and 3 of the six-member ring. Compounds with pyrimidine structures are known to possess antimicrobial1-2, antinflammatory3, cytotoxic4-5, anticancer activity6-7. In the present study some new pyrimidine derivatives (1 to 3) have been synthesized by the reaction of Aromatic Amines and Thiourea. The structures of the various synthesized compounds are assigned on the basis of elemental analysis, IR and 1HNMR Spectral data. These compounds were also screened for their anti-bacterial activity.

 

EXPERIMENTAL:

Step-1   Equimolar (0.02 mole) mixtures of Aromatic primary amines were refluxed in 40ml of dry ethanol for 18 hours. The excess of ethanol was then distilled off under reduced pressure and the resulting solid were washed with ethanol followed by ether. The compound was dried and Recrystallised from benzene.

 

Step-2    An equimolar (0.02moles) mixture of schiff’s base and Thioglycolic acid were refluxed for 18hr in 30ml of dry benzene. Then the excess benzene was distilled off under reduced pressure and the contents were poured over crushed ice and centrifuged. The resulting solids were washed with ice-cold saturated solution of sodium bicarbonate followed by distilled water. Then this compound was recrystallised from methanol.

 

Step-3   Equimolar mixture of thiazolidinones, Aromatic aldehydes and anhydrous sodium acetate (0.082gm) in 20ml of glacial acetic acid was refluxed for 3hrs. Then the reaction mixture was concentrated, cooled and poured into ice cold water. The solid crystal was separated by filtration and the filtrate was washed with water then the compound was washed with water then the compound was dried and recrystalised from glacial acetic acid.

 

 

Step-4   Step-3 and Thiourea or urea was dissolved in ethanol (50ml). To this NaOH dissolved to minimum quantity of water was added. Then the mixture was refluxed for 6hrs and poured into 250ml of cold water. The obtained was filtered, washed with water, and crystallized from ethanol.

 

SCHEME

 

R = Cl, Br                            Ar = C6H4OH(o-), C6H4NO2(p-)

 

Antimicrobial activity:- Antibacterial and antifungal activities were studied by subjecting the compounds to pharmacological screening by standard procedures.

All the compounds synthesized in the present investigation were tested for their antimicrobial activity. The antimicrobial activities were tested on nutrient medium against Pseudomonas aeruginosa, Klebsella Pneumonia, Escherichia coli, Pseudomonas mirabilis8.

 

Preparation of nutrient as a medium:-

The Nutrient agar media was prepared by using the following Ingredient:-

1.         Peptone                                   20g

2.         Beef extract                            5g

3          Sodium chloride                      5g

4.         Agar                                        20g

5.         Distilled water up to               1000ml

 


Table-1 [Physical properties of 2(2’ substituted phenyl)-1(4’ substituted phenyl) Amines

S. No.

Molecular structure

Molecular Formula

Molecular   Weight

M. P.

% Yield

Colour

Solubility

1.a

 

C13H10NOCl

231.5859

70 C

85

Orange Crystal

Benzene

1.b

 

C13H9ClN2O2

260.5

119 C

69

Yellow Crystal

Benzene

1.c

 

C14H12BrNO

290

76 C

75

Greenish grey crystal

Benzene

 

 

 

Table-2- [Physical properties of [2(substituted phenyl)-3(substituted phenyl) Thiazolidine-4-one]

S. No.

Molecular Structure

Molecular Formula

Molecular Weight

M. P.

% Yield

Colour

Solubility

 

2.a

 

C15H12ClNO2S

305.56

101 C

65.24

Orange  Crystal

Methanol

 

2.b

 

C15H11ClN2O3S

334.56

96 C

75

Yellowish

Methanol

 

 

2.c

 

C15H11BrN2O3S

379.06

133 C

88.5

Gray

Methanol

 

 

 

Table-3- [Physical properties of Knoveneagel reaction [2(substituted phenyl)-3(substituted phenyl)-5 (Substituted Benzylidine) Thiazolidine-4-one]

S. No.

Molecular Structure

Molecular Formula

Molecular Weight

M. P.

% Yield

Colour

Solubility

3.a

 

C22H16NO3SCl

409.9399

93 C

48

Yellowish gray

Glacial acetic acid

3.b

 

C22H15N2O4SCl

438.056

133 C

60

Grayish yellow

Glacial acetic acid

3.c

 

C22H15N2O4SBr

483.7273

70 C

41

Yellowish

Glacial acetic acid

 

Table-4- [Physical properties of step-lV]

S. No.

Molecular Structure

Moleculr Formula

Molecular Weight

M. P.

% Yied

Colour

Solubiliy

 

4.a

 

C22H18N3O2S2Cl

457.189

98 C

52

Yellowish

Ethanol

 

 

4.b

 

C23H17N4O2S2Cl

481.0938

125 C

62

Brownish yellow

Ethanol

 

 

 

4.c

 

C23H17N4O3S2Br

542.7178

103  C

68

Grayish yelloow

Ethanol

 

Table – V Spectral Data of the Compound (1 to 3)

Compounds

IR (KBr, cm-1)

1H NMR

4. (a)

3427.12(ArOH), 1611.22(C=N), 1360.12 (C-N), 1272.57(ArNH2), 1150.38(C=S), 757.65(C-Cl)

4.09, 5.48[C-H(S)], 1.2(S-H), 5.69,5.69(Phenolic OH bulk aromatic), 6.73-7.47(Proton)

4. (b)

3419.19(ArOH),1339.36(C-N),1397.46(ArNO2),1089.96(C=S),667.64(C-Cl)

4.09,5.48[C-H(S)],1.2(S-H),5.69(Phenolic OH bulk aromatic),7.05-7.69(Proton)

4. (c)

3432.50(ArOH), 1600.71(C=N), 1309.23(C-N), 1175.33(C=S), 619.19(C-Br)

4.09,5.48{C-H(S)},1.32(S-H),5.69(Phenolic OH bulk aromatic),7.05-7.69(Proton)

 

 


Weighed quantities of peptone, beef extract were dissolved in distilled water and pH was adjusted to 7.2-7.4 using pH paper. Then the specified amount of agar was added kept the beaker on hot water bath and allowed the agar to melt, it was dispensed in suitable containers and plugged them with non-adsorbent cotton they were sterilized by autoclave at 121c for20 minutes.

 

Preparation of solution of test compounds:-

Now 1mg/ml concentration of the test compounds was prepared using DMF as solvent it is chisonsidered, as stock solution from this 100g/ml concentration is prepared using DMF as solvent and this was used for antimicrobial activity studies9-10.

 

Preparation Standard Antibiotic Solution:-

Ampicillin 20 was used as a standard antibiotic for comparison and it was prepared by using sterile water.

Procedure:-  Sterile nutrient agar medium was cooled to 45 c this media was inoculated with 18-24 hours old bacterial culture under aseptic conditions mixed well by gentle shaking then it was poured in to sterile Petri dishes and allowed the medium to set. After setting all the seeded Petri dishes were transferred to laminar flow unit and 5 cups were made by using sterile cork borer. Out of 5 cups, 2 cups were added with 50 of the standard antibiotic (Ampicillion) and solvent control one in each bore, test compounds were added to the remaining 3 bores one in each bore. Then they were allowed for diffusion for 2 hours and incubated at 37ᵒC for 24hrs11-12. The inhibition zone diameters were measured and the results are shown in table-

 

 

S. No.

Compounds Code

Molecular Formula

P. Aurogenosa

K. Pneumonia

E.-Coli

P. Mirabilish

1.

4a

C22H18N3O2S2Cl

16

17

18

16.5

2

4b

C23H16N4O4S2Cl

15

13

14

13

3.

4c

C22H17N4O3S2Br

14

13

15

16

Bacteria used-

Pseudomonas aeruginosa,           Klebsella Pneumonia

Escherichia coli,        Pseudomonas mirabilis

 

RESULT AND DISCUSSION:

The resulted compounds were screened for antibacterial activity studies at a concentrated of 1.00 mg/ml using DMF as a control against P. Aurogenesa, K. Pneumonia, E-coli, P. Mirabilish by cup-plate method on nutrient agar Himedia, ampicillin 20 µg/ml used at standard against Gram positive and Gram negative bacteria. The data in table indicate that (4a1 and 4a2) compounds were found to passes a broad-spectrum activity, while other compounds were found to exhibit moderate activity. From this antibacterial screening it was found that the compounds showed significant activity and moderate activity at the given concentration levels. Perhaps pyrimidine base {2-(substituted phenyl)-3-(substituted phenyl) 2, 3, 7,7a-Tetrahydro [1, 3] thiazolo [4, 5 d] pyrimidine-5-(substituted) thione} may be largely responsible for the marked bactericidal.

The above results established the fact that the compound {2-(substituted phenyl)-3-(substituted phenyl) 2, 3, 7,7a- Tetrahydro [1, 3] thiazolo [4, 5 d] pyrimidine-5- (substituted thione} can be studied for the further investigation to search for the new antimicrobial compound.

 

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Received on 29.12.2010       Modified on 23.01.2011

Accepted on 27.01.2011      © RJPT All right reserved

Research J. Pharm. and Tech. 4(4): April 2011; Page 624-628