WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

Volume 3, Issue 3, 1981-1988. Research Article ISSN 2278 – 4357

SYNTHESIS AND ANTIMICROBIAL EVALUATION OF NEW

4{[(ARYL)METHYLENE]AMINO}-2-METHYL-5,6-SUBSTITUTED
THIENO[2,3-D]PYRIMIDINES

K. Naresh*1, Y. Rajeshwar2 and K. N. Jayaveera3

1
Jawaharlal Nehru Technological University Anantapur, Ananthapuramu, Andhra Pradesh,
India.
2
S. R. College of Pharmacy, Ananthasagar, Warangal, Andhra Pradesh, India.
3
Science tech foundation, Bangaluru, Karnataka, India.

Article Received on
12 February 2014,
Revised on 28 February 2014
Accepted on 15 March 2014
*Correspondence for Author
Naresh K
Jawaharlal Nehru
Technological University
Anantapur, Ananthapuramu,
Andhra Pradesh, India.
ABSTRACT
In continuation to our earlier reported work on the synthesis of novel
fused thieno[2,3-d]pyrimidine derivatives, we synthesized two new
series of 4{[(aryl)methylene]amino}-2-methyl-5,6-substituted
thieno[2,3-d]pyrimidines by the condensation of 4-amino-2,5,6–
substituted theino[2,3-d]pyrimidines (ANP and ANH) with nine
aromatic aldehydes. The synthesized compounds were characterized by
physical and spectral data. The compounds were further evaluated for
the antibacterial and antifungal activity employing disc diffusion
method.

Key words: thieno[2,3-d]pyrimidines, antibacterial, thiophene,

aromatic aldehydes

INTRODUCTION
The polysubstituted thiophenes especially when fused to pyrimidine ring; the
thienopyrimidine possess a large number of biological and pharmacological activities.
Thieno[2,3-d]pyrimidines as an isoster of purine with enhanced aromatic character; found to
have a wide variety of biological properties, as an agonists and antagonists to many receptors,
as inhibitors of enzymes such as phospodiesterases[1], VEGFR[2] and ErbB kinases[3]. Some of
the derivatives of thieno pyrimidines were also reported for activities such as antitumor[4],
antioxidant[5] and antibacterial[6]. The interesting chemical and biological profile of
thienopyrimidine prompted us to synthesize novel derivatives of it. We previously reported

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K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

some novel theno[2,3-d]pyrimidine derivatives for antibacterial, antifungal and antioxidant

activities[7]. In the present work we synthesized two series of new 2-methyl substituted
pentenyl/ heptenyl fused thienopyrimidine Schiff bases and evaluated for antibacterial and
antifungal activities.

EXPERIMENTAL
Structures of synthesized compounds were determined from their IR (Shimadzu FTIR), 1H
NMR (Gemini 300 MHz) and elemental analysis (Carlo ebra 1108 elemental analyzer) was
within ± 0.4% of theoretical value. Melting points of all the compounds were determined in
open capillaries using Toshniwal and Cintex melting point apparatus and are uncorrected.
Analytical TLC was performed on Silica Gel F 254 plates (Merck) and the spots were
visualized by UV or iodine vapors.

General procedure for the preparation of 4{[(aryl)methylene]amino}-2-methyl-6,7-

dihydro-5H-cyclopenta/6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidine
derivatives (ANP1-9 and ANH1-9)
2-methyl-6,7-dihydro-5H-cyclopenta/6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-
d]pyrimidine-4-amines (ANP and ANH) were prepared as per our previously reported
procedure. 0.001 mole of the each of the amines were further refluxed with aromatic
aldehyde in absolute ethanol with catalytic amount of glacial acetic acid for about 17 h. The
reaction mixture was then cooled in an ice bath and the crude product thus obtained was
collected by filtration. The crude compound was further purified by recrystallization from
ethanol.

4{[(phenyl)methylene]amino}-2-methyl-6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]
pyrimidine (ANP1)
IR (KBr): cm-1 2924.5 (C-H), 1644.6 (C=N); 1H NMR (CDCl3/ 300 MHz): 8.36( s, 1H, CH
of imino), 7.83 (m, 2H, Ar-H), 7.51 (m, 3H, Ar-H), 2.98 (m, 2H, CH2), 2.85 (m, 2H, CH2),
2.45 (s, 3H, CH3), 2.38 (m, 2H, CH2). Mass: m/z 293.

4{[(4-chlorophenyl)methylene]amino}-2-methyl-6,7-dihydro-5H-cyclopenta[4,5]thieno
[2,3-d]pyrimidine (ANP2)
IR(KBr): cm-1 2946.72 (C-H), 1647.30 (C=N); 1H NMR (CDCl3/ 300 MHz): 8.36 ( s, 1H, CH
of imino), 7.75 (d, 2H, Ar-H), 7.52 (d, 2H, Ar-H), 2.98 (m, 2H, CH2), 2.85 (m, 2H, CH2),
2.45 (s, 3H, CH3), 2.39 (m, 2H, CH2); Mass: m/z 327.

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K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

4{[(phenyl)methylene]amino}-2-methyl-6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno
[2,3-d]
Pyrimidine (ANH1)
IR(KBr): cm-1 2946.72 (C-H), 1647.30 (C=N); 1H NMR (CDCl3/ 300 MHz): 8.36 ( s, 1H, CH
of of imino), 7.83 (m, 2H, Ar-H), 7.51 (m, 3H, Ar-H), 2.85 (m, 2H, CH2), 2.55 (m, 2H, CH2),
2.45 (s, 3H, CH3), 1.84 (m, 2H, CH2), 1.72 (m, 4H, CH2); Mass: m/z 321.

4{[(4-N,N-dimethylaminophenyl)methylene]amino}-2-methyl-6,7,8,9-tetrahydro-5H-
cyclohepta[4,5]thieno[2,3-d]pyrimidine (ANH4)
IR(KBr): cm-1 2950.60 (C-H), 1623.38 (C=N); 1H NMR (CDCl3/ 300 MHz): 8.36 ( s, 1H, CH
of imino), 7.50 (d, 2H, Ar-H), 6.80 (d, 2H, Ar-H), 3.06 (s, 6H, N(CH3)2), 2.85 (m, 2H, CH2),
2.55 (m, 2H, CH2), 2.44 (s, 3H, CH3), 1.84 (m, 2H, CH2), 1.72 (m, 4H, CH2); Mass: m/z 364.

Table 1. Physical data of synthesized compounds

Substituents Molecular Melting point Yield

Compound
R1 R2 Ar weight (gm) (⁰C) (%)
ANP1 -(CH2)3- -C6H5 293 170-171 80
ANP2 -(CH2)3- p-Cl-C6H4 328 156-158 83
ANP3 -(CH2)3- p-Br-C6H4 372 193-196 66
ANP4 -(CH2)3-- p-N(CH3)2-C6H4 336 165-167 75
ANP5 -(CH2)3- p-N(C2H5)2-C6H4 365 110-112 71
ANP6 -(CH2)3- p,m-(OCH3)2-C6H3 353 178-180 63
ANP7 -(CH2)3- p-(OCH3)-C6H4 323 155-158 69
ANP8 -(CH2)3- 3-indolyl 332 149-151 77
ANP9 -(CH2)3- 3-thienyl 299 112-114 61
ANH1 -(CH2)5- -C6H5 321 152-154 86
ANH2 -(CH2)5- p-Cl-C6H4 356 173-175 79
ANH3 -(CH2)5- p-Br-C6H4 400 190-192 65
ANH4 -(CH2)5- p-N(CH3)2-C6H4 365 175-177 83
ANH5 -(CH2)5- p-N(C2H5)2-C6H4 393 165-168 85
ANH6 -(CH2)5- p,m-(OCH3)2-C6H3 381 149-152 71
ANH7 -(CH2)5- p-(OCH3)-C6H4 351 163-166 81
ANH8 -(CH2)5- 3-indolyl 360 170-172 79
ANH9 -(CH2)5- 3-thienyl 327 196-198 62

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K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

O R 1 CN
R 1 CN
S u lp h u r
+ H 2C
M o r p h o li n e
CN R NH 2
R 2 S
2
C y c lo p e n ta n o n e / P and H
C y c lo h e p ta n o n e
M a le n o n itrile

N aO C H 3 A c e to n itrile

NH 2
R 1

N
R 2
S N R 3

A N P an d A N H

R 1 = R 2 = - ( C H 2 ) 3 , - ( C H 2 )5
R 3 = -C H 3
E th a n o l A r-C H O
A c e t i c a c id

Ar = C 6 H 5 , p - C l- C 6 H 4 , p -B r - C 6 H 4 ,
p - N ( C H 3 ) 2 C 6H 4 , p -N ( C 2 H 5 )C 6 H 4 ,
p ,m - ( O C H 3 ) 2 -C 6 H 3 , p - ( O C H 3 ) - C 6 H 4 ,
3 - i n d o ly l , 3 - T h i e n y l CH Ar
N
R 1

N
R 2
S N R 3

A N P 1-9 a n d A N H 1-9

Scheme-I
BIOLOGICAL ACTIVITY
Antibacterial activity
All the newly synthesized 4{[(aryl)methylene]amino}-2-methyl-6,7-dihydro-5H-cyclopenta/
6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidine derivatives (ANP1-9 and
ANH1-9) were evaluated for antibacterial activity against S.aureus, B.subtilis, E.coli and
P.aeruginosa by employing the disc diffusion method[8]. The antibacterial activity of each of
the test compounds was carried out at 500 µg/ ml and 1000 µg/ ml concentrations in DMSO.
Streptomycin was used as standard drug to compare the antibacterial activity. The diameter of
the zone of inhibition of the microorganism caused by the test and standard compounds was
measured in mm. All tests were carried out in triplicate and results are presented as mean and
standard deviation in table 2.

Antifungal activity
All the synthesized 4{[(aryl)methylene]amino}-2-methyl-6,7-dihydro-5H-cyclopenta/
6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidine derivatives were also screened
for antifungal activity against Aspergillus fumigatus, Aspergillus flavus, Candida albicans
and Candida krusei. Disc diffusion method[9, 10] was used for the antifungal study as per the
reported procedures. Test dilutions 500 µg/ ml and 1000 µg/ ml were prepared for each of the
test compounds in DMSO and clotrimazole was taken as standard drug. All tests were
performed in triplicate and the diameter of the zone of inhibition was recorded and results are
presented as mean and standard deviation in table 3.

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K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

Table 2. Anti bacterial activity of new 4{[(aryl)methylene]amino}-2-methyl-6,7-dihydro-

5H-cyclopenta/6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidine derivatives
(ANP1-9 and ANH1-9)

Diameter of zone of inhibition (mm) Mean ± SD

Compound
P. aeruginosa E. coli S. aureus B. subtilis
500 µg - 13±0.4 14±1.0 12±0.3
ANP1
1000 µg 16±0.4 16±0.7 16±0.4 17±0.4
500 µg 16±0.5 16±0.5 16±0.5 15±0.5
ANP2
1000 µg 19±0.2 19±0.5 20±0.6 19±0.4
500 µg 12±0.4 14±0.9 13±0.3 12±0.4
ANP3
1000 µg 18±0.3 18±0.3 16±0.9 14±0.9
500 µg 13±0.5 14±0.4 14±0.5 14±0.8
ANP4
1000 µg 17±0.1 16±0.9 19±0.3 16±1.0
500 µg 16±0.4 16±0.4 16±0.7 15±0.3
ANP5
1000 µg 19±0.4 17±0.6 18±0.4 19±0.7
500 µg 12±0.5 15±0.8 14±0.7 13±0.8
ANP6
1000 µg 17±0.6 18±0.4 18±0.7 16±0.6
500 µg 14±0.3 14±0.7 14±0.8 13±0.6
ANP7
1000 µg 19±0.4 19±0.4 16±1.5 18±0.4
500 µg 15±0.5 14±0.8 16±1.1 16±0.4
ANP8
1000 µg 19±0.3 21±0.3 16±0.3 18±0.3
500 µg 16±0.6 15±0.4 16±0.4 15±0.5
ANP9
1000 µg 19±0.5 21±0.2 20±0.3 21±0.6
ANH1 500 µg 13±1.9 14±0.6 13±0.3 12±0.6
1000 µg 16±0.8 18±0.8 16±0.9 17±0.7
ANH2 500 µg 15±0.7 14±0.6 16±0.7 15±0.7
1000 µg 19±0.5 21±0.5 20±0.6 21±0.4
ANH3 500 µg 13±0.8 13±0.7 12±0.8 11±0.6
1000 µg 18±0.8 16±0.7 13±0.3 17±0.5
ANH4 500 µg 14±0.3 13±0.3 13±1.7 13±0.6
1000 µg 17±0.5 19±0.6 16±1.5 17±0.4
ANH5 500 µg - 14±0.5 14±1.0 12±0.9
1000 µg 18±0.7 19±0.8 15±0.7 15±0.6
ANH6 500 µg 12±0.6 13±0.4 13±0.4 14±0.8
1000 µg 16±0.5 16±0.8 15±0.6 16±0.4
ANH7 500 µg 14±0.6 13±0.8 13±0.6 12±0.3
1000 µg 14±0.4 18±0.4 15±0.8 16±0.7
ANH8 500 µg 16±0.6 14±0.6 15±0.5 16±0.8
1000 µg 19±0.8 19±0.5 17±0.9 18±0.5
ANH9 500 µg 13±0.8 14±0.7 14±1.1 13±1.9
1000 µg 17±0.7 18±0.9 17±0.7 18±0.9
S* 100 µg 23±0.4 24±0.1 23±0.4 24±0.1
S* = Streptomycin

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K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

Table 3. Antifungal activity of new 4{[(aryl)methylene]amino}-2-methyl-6,7-dihydro-

5H-cyclopenta/6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidine derivatives
(ANP1-9 and ANH1-9)
Diameter of zone of inhibition(mm) Mean ± SD
Compound
A. fumigatus A. flavus C. albicans C. krusei
500 µg 12±0.7 - 12±0.3 12±0.7
ANP1
1000 µg 15±0.8 13±0.3 13±0.5 14±0.8
500 µg 12±0.7 13±0.5 13±0.6 12±0.3
ANP2
1000 µg 16±0.6 13±0.6 15±0.7 15±0.5
500 µg 12±0.5 12±0.6 13±0.8 12±0.5
ANP3
1000 µg 16±0.8 14±0.5 13±0.8 16±0.4
500 µg 12±0.7 13±0.4 12±0.8 13±0.6
ANP4
1000 µg 12±0.5 15±0.7 15±0.5 14±0.5
500 µg 14±0.4 12±0.5 13±0.7 12±0.5
ANP5
1000 µg 14±0.6 16±0.7 14±0.7 14±0.2
500 µg 13±0.5 12±0.4 12±0.3 12±0.6
ANP6
1000 µg 15±0.7 18±0.8 14±0.5 14±0.6
500 µg 13±0.6 12±0.6 11±0.8 12±0.7
ANP7
1000 µg 14±0.7 16±0.3 15±0.8 14±0.5
500 µg 12±0.5 12±0.7 13±0.2 11±0.6
ANP8
1000 µg 13±0.3 14±0.8 14±0.4 14±0.8
500 µg 12±0.5 11±0.6 13±0.4 12±0.6
ANP9
1000 µg 15±0.7 16±0.5 15±0.5 16±0.7
500 µg 12±0.3 13±0.8 12±0.6 12±0.8
ANH1
1000 µg 14±0.8 16±0.9 16±0.7 15±0.7
500 µg 12±0.6 12±0.7 12±0.7 12±0.7
ANH2
1000 µg 15±0.5 14±0.4 14±0.7 14±0.7
500 µg 13±0.7 12±0.9 13±0.8 13±0.8
ANH3
1000 µg 16±0.4 16±0.3 16±0.8 17±0.7
500 µg 14±0.4 14±0.4 13±0.9 14±1.0
ANH4
1000 µg 18±0.5 18±0.5 15±0.9 16±1.2
500 µg 12±0.5 14±0.4 13±0.5 12±0.9
ANH5
1000 µg 18±0.6 18±0.6 17±0.5 18±0.7
500 µg 13±0.4 13±0.6 12±0.5 12±0.7
ANH6
1000 µg 18±0.4 18±0.7 18±0.7 19±0.7
500 µg 12±0.3 12±0.6 12±0.7 14±0.8
ANH7
1000 µg 18±0.7 14±0.3 16±0.5 16±0.4
500 µg 15±0.3 15±0.9 12±0.9 12±0.8
ANH8
1000 µg 20±0.7 17±0.6 18±0.9 18±0.3
500 µg 12±0.3 13±0.7 15±0.1 12±0.4
ANH9
1000 µg 17±0.9 18±0.6 16±1.8 18±0.8
S* 100 µg 24±0.4 20±0.5 22±0.3 23±0.5

S* = Clotrimazole

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K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

RESULTS AND DISCUSSION

Two series of new 4{[(aryl)methylene]amino}-2-methyl-5,6 substituted thieno[2,3-
d]pyrimidine (ANP1-9 and ANH1-9) were synthesized by condensing 2-methyl-6,7-dihydro-
5H-cyclopenta/6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidine-4-amines
(ANP and ANH) with nine aromatic aldehydes benzaldehyde, p-chloro benzaldehyde, p-
bromobenzaldehyde, p-N,N-dimethyl amino benzaldehyde, p-N,N-diethyl amino
benzaldehyde, p-methoxy benzaldehyde, 3,4-dimethoxy benzaldehyde, indole-3-
carboxaldehyde and thiophene-3-carboxaldehyde. The thieno[2,3-d]pyrimidine-4-amines
were prepared by following our earlier reported procedures (scheme-1) using cyclopentanone
and cycloheptanone as ketones.

The antibacterial activity of the synthesized compounds revealed that the compounds ANP2,
ANP5, ANP9 and ANH2 showed highest activity on all the tested strains of bacteria than the
other compounds of the two series. However, compounds ANP3, ANP7, ANP8, ANH4,
ANH6, ANH8 and ANH9 exhibited appreciable antibacterial activity; whereas compounds
ANP1, ANP4, ANP6, ANH1, ANH3, ANH5 and ANH7 showed a considerable antibacterial
activity. It was also noticed that most of the tested compounds were found to be active
against E.coli and P.aeruginosa and none of them were as active as standard drug
streptomycin at the tested concentrations.

All the tested compounds were evaluated for antifungal activity and from the results, it was
observed that compounds ANP6 and ANH4 exhibited highest activity against Aspergillus
species; whereas, the activity is more on Candida species with the compounds ANP2, ANP9
and ANH6. All the remaining compounds except ANH1 and ANH2 exhibited a considerable
activity when compared with the standard drug clotrimazole.

CONCLUSION
In view of the reported interested biological properties of thienopyrimidines, two series of
4{[(aryl)methylene]amino}-2-methyl-5,6 substituted thieno[2,3-d]pyrimidine (ANP1-9 and
ANH1-9) were prepared and screened. From the antibacterial and antifungal investigations of
these derivatives, it could be concluded that the compounds ANP2, ANP5, ANP9 and ANH2
are valuable scaffolds for potential antibacterial agents, whereas compounds ANP2, ANP6,
ANP9, ANH4 and ANH6 are scaffolds for the future antifungal agents.

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K. Naresh et al. World Journal of Pharmacy and Pharmaceutical Sciences

ACKNOWLEDGMENTS
Authors are thankful to the Director, Research and Development, JNTUA, Ananthapuramu,
India. Authors are also thankful to Director, CSIR-IICT, Hyderabad, India for the spectral
data.

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