The utility of carbon disulphide and lawesson's reagent for synthesis of different fused heterocycles for antimicrobial evaluation

Article abstract of DOI:10.1002/jhet.884

Hexahydroquinolines 1a, 1b reacted with carbon disulphide in different conditions to yield the corresponding adducts 2a, 2b and 3a, 3b. Carrying out the same reactions in acetone as solvent produced the modified new products 4a, 4b. The interaction of pyr

Full text of DOI:10.1002/jhet.884

Month 2013
The Utility of Carbon Disulphide and Lawesson's Reagent
for Synthesis of Different Fused Heterocycles for
Antimicrobial Evaluation
a
b
c
Nadia R. Mohamed, * Nahed Y. Khaireldin, Amin F. Fahmy,
b
and Ahmed A. El-Sayed
a
Chemistry Department, El-Aflaj Girls College, El-Kharj University Kingdom of Saudi Arabia
b
Photochemistry Department, National Research Centre, Dokki, Giza, Egypt
c
Chemistry Department Faculty of Science, Ain Shams University, Cairo, Egypt
*
E-mail: nadia_ragab_m@hotmail.com
Received November 26, 2010
DOI 10.1002/jhet.884
Published online 00 Month 2013 in Wiley Online Library (wileyonlinelibrary.com).
Hexahydroquinolines 1a,b reacted with carbon disulphide in different conditions to yield the
corresponding adducts 2a,b and 3a,b. Carrying out the same reactions in acetone as solvent produced
the modified new products 4a,b. The interaction of pyrazolopyridine derivatives 5a–d with carbon
disulphide under the same previous conditions furnished the isolated products 6a–d, 7a–d, and 8a–d.
Studying the behavior of 1a,b or 5a–d toward Lawesson's reagent (LR) formed the final adducts 11a,b
or 12a–d. The structure of synthesized compounds was confirmed with the spectroscopic and microana-
lytical data. The biological activities of 2a, 4a, 4b, 7a, 7c, 8d, 11a, 11b, 12b, and 12c were tested for
antimicrobial evaluation.
J. Heterocyclic Chem., 00, 00 (2013).
INTRODUCTION
heterocycles [14–17], we explain in this report the
reaction of carbon disulphide and LR with
The pyrazolo and pyranopyridopyrimidine derivatives
play an important role in biochemical processes because
the side groups of the most typical and essential constit-
uents of living cells DNA and RNA are based on
aromatic heterocycles [1,2]. They are known to exhibit
pharmacological activates such as CNS depressant,
neuroleptic, and turberculostatic [3–5]. Heterocyclic
compounds containing sulfur, phosphorus, and nitrogen
have maintained the interest of researchers through
decades of historical development of organic synthesis
hexahydroquinolines
1a,
pyrazolopyranes,
and
pyrazolopyridines 5a–d. Different bioactive fused het-
erocycles were synthesized and evaluated as antimicro-
bial agents.
RESULTS AND DISCUSSION
2-Amino-3-cyano-hexahydroquinoline derivatives 1a,b
[18] were allowed to react with carbon disulphide under
different reaction conditions. Interaction of 1a,b with
equivalent amount carbon disulphide (1:1 mole) in alco-
holic potassium hydroxide 10% yielded the corresponding
adducts 2a,b (Scheme 1). The IR spectrum of 2a, taken as
[
6–10]. On the other hand, carbon disulphide and
Lawesson's reagent (LR) have been developed as
powerful and versatile reagents in organic synthesis
[
11–13]. Continuation to our previous work for using
these reagents to synthesize different bioactive
©
2013 HeteroCorporation

N. R. Mohamed, N. Y. Khaireldin, A. F. Fahmy, and A. A. El-Sayed
Vol 000
Scheme 1
produced the final product 4a,b. Similar mechanism was
reported previously which support our suggested point of
view [19] (Scheme 2).
To synthesize other different types of sulfur fused
heterocycles, the previous reactions were taken by using
pyrazolopyridines and pyrazolopyranes 5a–d [20] as
starting materials. Interaction of 5a–d with carbon
disulphide under the same different previous conditions
yielded the corresponding adducts 6a–d, 7a–d, and 8a–d
(Scheme 3). The structure of all adducts were elucidated
according the microanalytical and spectroscopic data (c.f.,
Experimental section).
Treatment of 1a,b with 0.05 mol of LR in dry toluene un-
der reflux led to the formation of the addition intermediated
9
a,b which cyclized to 10a,b under the reaction conditions.
Rearrangement of 10a,b produced the final isolated product
1a,b [21] (Scheme 4). The IR spectrum of 11a revealed the
absence of the NH and CN groups of 1a and the presence of
1
2
−1
the NH at v =3289–3185 cm (Scheme 4).
example, revealed the presence of three NH groups at v =
−
1
3
1
281–3190 cm and two carbonyl groups at v = 1690,
−
1 1
665 cm . H-NMR spectrum showed the presence of
three NH groups, D O exchangeable (c.f., experimental
2
section). Carrying out the previous reaction by using
excess of carbon disulphide furnished the corresponding
products 3a,b (Scheme 1). The structure of latter com-
pounds was confirmed with mass spectrum which revealed
+
m/z [M ] at 369 and 405, and the other spectroscopic data
were in accordance with the suggested structures. Aiming
to study the effect of solvent on the reaction's type of prod-
ucts, the reaction was allowed to take place in acetone as
solvent, and the formed products in this case were 4a,b
(
Scheme 1). The elemental analysis of 4a,b proved the
1
absence of sulfur and the H-NMR of 4a spectrum showed
the presence of the original two methyl groups at δ = 0.86
and 1.01 ppm and the presence of other new methyl groups
at δ = 1.95 and 2.01 ppm. The C-NMR spectrum
supported the presence of the four methyl groups at δ =
Figure 1. Molecular structure of the product 4a or b in the crystal; the
crystallographic numbering does not represent the systematic numbering.
Intermolecular bond lengths and bond angles are shown. Selected bond
lengths [A°] and bond angles [°] limits use covalent radii + 0.020 A°.
1
3
2
8.68 and 30.86 ppm.
One crystal X-ray pattern of 4a was made and supported
[Color figure can be viewed in the online issue, which is available at
wileyonlinelibrary.com.]
our suggested structure under number CCDC 796433
(
Fig. 1).
To specify the role of carbon disulphide in the later reac-
On the other hand, 5a–d was reacted with LR under the same
previous condition to form the final products 12a–d (Scheme 5).
The suggested structure of 11a,b and 12a–b was confirmed
with different spectroscopic data (c.f., Experimental section).
Antimicrobial activity. Evaluation of the antimicrobial
activity of compounds 2a, 4a, 4b, 7a, 7c, 8d, 11a, 11b,
12b, and 12c were performed at the Microanalytical
Center, Faculty of Science, Cairo University. The tested
compounds were evaluated for their activities against the
Gram positive, Gram negative bacteria, and fungi. Tested
tion's mechanistic sequence, the reaction was carried out in
absence of carbon disulphide. It recovered in this case the
starting material 1a which mean the importance of carbon
disulphide for formation of the product. The expected
mechanism of the reaction is the attack of carbon
disulphide on the amino group of 1a,b at first to form the
intermediate A. The later hydrolyzed and reacted with
one molecule of acetone to form the intermediate B. Cycli-
zation of B with loss of carbon disulphide and water
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2013
The Utility of Carbon Disulphide and Lawesson's Reagent for
Synthesis of Different Fused Heterocycles for Antimicrobial Evaluation
Scheme 2
Scheme 3
CONCLUSIONS
In summary, the synthesized compounds of different fused
heterocyclic derivatives exhibit promising antimicrobial ac-
tivity. Substitution of halo, phosphorous, and sulfurs groups
emerged as active in both antibacterial and antifungal screen-
ing. Moreover, this preliminary study is encouraging to fur-
ther explore their broad spectrum pharmacological
activities. The results of antibacterial and fungi studies are
given in Table 1. Compound 8d was showed complete
inhibition at 30 mm/mg for Escherichia coli (Gram negative
bacteria) and Staphylococcus aureus (Gram positive bacteria)
and showed inhibition at 28 mm/mg higher than the
Amphotericin B as antifungal standard for the Candida
albicans (Fungus). Compounds 4b and 11b showed moder-
ate inhibition for the two positive and negative bacteria
between 22 and 14 mm/mg. The rest of the tested com-
pounds 7a, 11a, and 13b showed moderate inhibition for bac-
teria between 17 and 13 mm/mg and have 15–11 mm/mg
effect on fungi. On the other hand, compound 2a showed
low antibacterial inhibition for the two at 10 mm/mg.
EXPERIMENTAL
Melting points were determined on an electrothermal apparatus
(
Buchi 535, Switzerland) in an open capillary tube and are
−
1
uncorrected. IR spectra expressed in (cm ) were recorded in
1
KBr pellets on a PA-9721 IR spectrophotometer. H-NMR and
1
3
C-NMR spectra were obtained on a Varian EM-390 (270 and
5
00 MHz) spectrometer in DMSO-d as solvent, using TMS as
6
internal reference and chemical shifts (δ) are expressed in ppm.
Mass spectra were recorded on Kratos (75 eV) Ms Equipment.
Elemental analyses were carried out by the microanalytical unit at
National Research Centre, Giza, Egypt. All reactions were moni-
tored by thin layer chromatography, carried out on 0.2 mm silica
compounds were added to the monolayer of Gram positive,
Gram negative bacteria, and fungi. A 48 h continuous drug
exposure was used to estimate all availability or growth.
Table 1 represents the activity for each compound.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

N. R. Mohamed, N. Y. Khaireldin, A. F. Fahmy, and A. A. El-Sayed
Vol 000
Scheme 4
Scheme 5
8,8-Dimethyl-5-phenyl-2-thioxo-1,3,5,7,9,10-hexahydropyrimido
4,5-b]quinoline-4,6-dione (2a). Yellow crystals, mp 246–247°C,
[
+
−1
(
3
EtOH), yield (77%); Ms. m/z: 353 [M , 45%]. IR (KBr) v cm :
1
3
281–3190 (NH), 2956 (CH ), 1690 (C¼O), 1665 (C¼O). H-
6 3
NMR (270 MHz, DMSO-d , TMS): δ 0.86 (s, 3H, CH ), 1.01 (s,
3
H, CH ), 1.95 (s, 2H, -CH ), 2.29 (s, 2H, -CH ), 4.81 (s, 1H, -
3 2 2
CH), 7.03–7.16 (m, 6H, 5 aromatic protons and NH, D
exchangeable), 8.52 (s, 1H, NH, D O exchangeable), 12.30 (s,
H, NH, D O exchangeable). Anal. Calcd. For Molecular
Formula C H N O S: C, 64.57%; H, 5.38%; N, 11.89%;
2
O
2
1
2
19 19 3 2
Found: C, 64.19%; H, 5.01%; N, 11.60%.
-(4-Chlorophenyl)-8,8-dimethyl-2-thioxo-1,3,5,7,9,10-
5
hexahydropyrimido[4,5-b]quinoline-4,6-dione (2b). Pale yellow
crystals, mp 290–292°C, (EtOH), yield (62%); Ms. m/z: 389
+
2
37
+
35
−1
[
3
(
M , Cl , 32%]; 387 [M , Cl , 15%]: IR (KBr) v cm : 3281–
1
195 (NH), 2985 (CH ), 1735 (C¼O), 1690 (C¼O). H-NMR
3
270 MHz, DMSO-d , TMS): δ 0.89 (s, 3H, CH ), 1.17 (s, 3H,
6
3
gel 60 F-254 (Merck) plates using UV light (245 and 365 nm) for
detection. Column chromatog-raphy was carried out on a Baker
silica gel powder (60–200 mesh).
CH
3
), 1.98 (s, 2H, -CH
2
), 2.26 (s, 2H, -CH
2
), 4.90 (s, 1H, -CH),
7
.13 (d, 2H, J = 8.9; aromatic), 7.28 (d, 2H, J = 8.9; aromatic),
.52 (s, 1H, NH, D O exchangeable), 10.32 (s, 1H, NH, D
8
2
2
O
General procedure: Synthesis of 2a,b and 6a–d. To a
solution of 1a,b or 5a–d (0.29 or 0.32 g, 0.001 mol), in 15 mL
of ethanol/KOH (10%), carbon disulphide (0.076 g, 0.001 mol)
was added. The reaction mixture was heated under reflux for 4-6 h.
The solvent was evaporated under vacuum, and the remaining
solids were crystallized with the proper solvent.
exchangeable), 11.85 (s, 1H, NH, D O exchangeable). Anal.
2
Calcd. For Molecular Formula C H N O ClS: C 58.83%; H,
19 18 3 2
4
8
.68%; Cl, 9.14%; N, 10.83%; Found: C, 58.43%; H, 4.38%; Cl,
.74%; N, 10.59%.
General procedure: Synthesis of 3a,b and 7a–d. To a
solution of 1a,b or 5a,b (0.29 or 0.32 g, 0.001 mol), in 15 mL
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2013
The Utility of Carbon Disulphide and Lawesson's Reagent for
Synthesis of Different Fused Heterocycles for Antimicrobial Evaluation
Table 1
Antimicrobial activity of compounds.
Inhibition zone diameter (mm/mg sample)
Escherichia
coli (G )
Staphylococcus
aureus (G )
Aspergillus
flavus (fungus)
Candida
albicans (fungus)
−
+
Sample
Solvent: DMSO
Standard
0.0
0.0
0.0
0.0
Tetracycline antibacterial agent
Amphotericin B antifungal agent
a
a
b
a
32
–
30
–
–
16
0.0
0.0
3
6
0.0
5
–
19
2
4
4
7
7
8
1
1
1
1
10
0.0
22
17
0.0
30
12
15
15
0.0
10
0.0
20
16
0.0
30
13
14
16
0.0
0.0
0.0
0.0
15
0.0
28
11
0.0
13
c
d
1a
1b
2b
2c
0.0
1
2
0.0
0.0
G: Gram reaction.
of ethanol/KOH (10%), excess of carbon disulphide was added.
The reaction mixture was heated under reflux for 3–6 h. The
solvent was evaporated under vacuum, and the remaining solids
were crystallized from the proper solvent.
General procedure: Synthesis of 4a,b and 8a–d. To a
solution of 1a,b or 5a–d (0.29 or 0.32 g, 0.001 mol), in 15 mL
of acetone/KOH (10%), excess of carbon disulphide was added
dropwise. The reaction mixtures were stirred under reflux until
no more of the starting materials were detected by TLC (3.5–5
h). The solvent was evaporated under vacuum, and the
remaining solid was crystallized with proper solvent.
8
,8-Dimethyl-5-phenyl-2,4-dithioxo-1,3,5,7,9,10-octahydro-
pyrimido[4,5-b]quinolin-6-one (3a). Pale yellow crystals, mp
+
+
2
40–241°C, (EtOH), yield (62%); Ms. m/z (M ); 369 [M , 50%].
−
1
IR (KBr) v cm : 3280–3202 (NH), 2950 (CH
3
), 1745 (C¼O).
, TMS): δ 0.86 (s, 3H, -CH ),
.01 (s, 3H, -CH ), 1.95 (s, 2H, -CH ), 2.21 (s, 2H, -CH ),
2
,2,8,8-Tetramethyl-5-phenyl-1,3,5,7,9,10-octahydropyrimido
1
H-NMR (500 MHz, DMSO-d
6
3
[
4,5-b]quinoline-4,6-dione (4a). Yellow crystals, mp 276–277°
+
1
4
3
2
2
C, (EtOH), yield (69.5%); Ms. m/z; 351 [M , 35%]. IR (KBr) v
−1
.80 (s, 1H, -CH), 7.01–7.35 (m, 6H, 5 aromatic protons and
O exchangeable), 9.32 (s, 1H, NH, D O exchangeable),
O exchangeable). C-NMR (500 MHz,
DMSO-d , TMS): 26.03 (C-8), 29.68 (2CH ), 30.26 (C-5),
cm : 3281–3186 (NH), 2956 (CH ), 1737 (C¼O), 1650 (C¼O).
3
1
NH, D
1.02 (s, 1H, NH, D
2
2
H-NMR (500 MHz, DMSO-d , TMS): δ 0.85 (s, 3H, -CH ),
6
3
1
3
1
2
1
.01 (s, 3H, -CH
3
), 1.95 (s, 3H, -CH
3
3
), 2.01 (s, 3H, -CH ), 2.25
6
3
2 2
(s, 2H, -CH ), 2.40 (s, 2H, -CH ), 4.80 (s, 1H, -CH), 7.16–7.60
(m, 6H, 5 aromatic protons and NH, D O exchangeable), 9.32 (s,
2
3
1
2.86 (2CH ), 50.10 (C-9), 53.32 (C-7), 52.60 (C-2), 92.83 (C-
1), 106.54 (C-13), 123.83, 128.69, 129.08, 127.75 (6C,
3
1
H, NH, D O exchangeable), 10.43 (s, 1H, NH, D O
2
2
13
benzene ring), 148.43 (C-14), 152.25 (C-12), 162.03 (C¼O),
74.67 (C¼S), 194.02 (C¼S). Anal. Calcd. For Molecular
Formula C H N OS : C, 61.76%, H, 5.18%, N, 11.37%;
exchangeable). C-NMR (500 MHz, DMSO-d , TMS): 26.53
6
1
(
C-8), 28.68 (2CH
3 3
), 29.26 (C-5), 30.86 (2CH ), 42.98 (C-9),
1
9
19
3
2
50.32 (C-7), 52.60 (C-2), 92.83 (C-11), 111.54 (C-13), 119.83,
Found: C, 61.28%, H, 4.95%, N, 11.21%.
-(4-Chlorophenyl)-8,8-dimethyl-2,4-dithioxo-1,3,5,7,9,10-
octahydro-pyrimido[4,5-b]quinolin-6-one (3b). Brown
crystals, mp 220–223°C, (EtOH), yield (69%); Ms. m/z; 405
1
1
25.69, 127.08, 127.75 (6C, phenyl), 146.43 (C-14), 147.25 (C-
5
2), 165.03 (C¼O), 194.67 (C¼O). Anal. Calcd. For Molecular
Formula Molecular C H N O : Composition = C, 71.77%; H,
21 25 3 2
7
.17%; N, 11.96%; Found: C, 71.45%; H, 6.95%; N, 11.72%.
-(4-Chlorophenyl)-2,2,8,8-tetra-methyl-1,3,5,7,9,10-octahy-
+
2
37
+
35
−1
[
3
M , Cl , 20%], 403 [M , Cl , 8%]. IR (KBr) v cm :
5
1
280–3186 (NH), 2960 (CH ), 1692 (C¼O). H-NMR (500
3
dropyrimido[4,5-b]quinolin-6-one (4b). Pale yellow crystals,
mp 295–296°C (decomposed), (EtOH), yield (80%); Ms. m/z
MHz, DMSO-d
2
1
7
6
, TMS): δ 0.85 (s, 3H, -CH
.01 (s, 2H, -CH ), 2.21 (s, 2H, -CH ), 4.45 (s, 1H, -CH), 6.70 (s,
H, NH, D O exchangeable), 7.13 (d, 2H, J = 9.1, 2H aromatic),
.26 (d, 2H, J = 9.1, 2H aromatic), 9.32 (s, 1H, NH, D O
3 3
), 1.00 (s, 3H, -CH ),
+
+2
37
+
35
2
2
(
M ); 387 [M , Cl , 21%], 385 [M , Cl , 10%]. IR (KBr) v
−
1
2
cm : 3280–3195 (NH), 2950 (CH
), 1757 (C¼O), 1670
3
1
2
(C¼O). H-NMR (500 MHz, DMSO-d , TMS): δ 0.85 (s, 3H, -
6
13
exchangeable), 10.92 (s, 1H, NH, D
2
O exchangeable). C-NMR
), 32.26
C-5), 35.86 (2CH ), 51.10 (C-9), 55.32 (C7), 56.60 (C-2), 92.83
CH ), 0.94 (s, 3H, -CH ), 1.94 (s, 3H, -CH ), 2.15 (s, 3H, -
3
3
3
(
500 MHz, DMSO-d
6
, TMS): 25.03 (C-8), 30.68 (2CH
3
CH ), 2.30 (s, 2H, -CH ), 2.45 (s, 2H, -CH ), 4.79 (s, 1H, CH),
6.45 (s, 1H, NH, D O exchangeable), 7.16 (d, 2H, J = 6.11;
3 2 2
(
3
2
(
C-11), 106.54 (C-13), 123.83, 128.69, 130.08, 135.75 (6C,
chlorobenzene), 148.43 (C-14), 152.25 (C-12), 161.03 (C¼O),
70.67 (C¼S), 191.02 (C¼S). Anal. Calcd. For Molecular Formula
Molecular C H ClN OS : C, 56.49%; H 4.46%; N, 10.40%; Cl
aromatic), 7.35 (d, 2H, J = 6.11, aromatic); 8.43 (s, 1H, NH, D O
2
13
exchangeable); 10.32 (s, 1H, NH, D O exchangeable). C-NMR
2
1
(500 MHz, DMSO-d , TMS): 26.53 (C-8), 28.68 (2CH ), 29.26
6
3
(C-5), 32.86 (2CH ), 42.98 (C-9), 50.32 (C-7), 52.60 (C-2), 92.83
19
18
3
2
3
8
.80%; Found: C, 56.09%, H, 4.13%, N, 9.95%; Cl, 8.46%.
(C-11), 111.54 (C-13), 120.83, 127.69, 130.08, 135.75 (6C,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

N. R. Mohamed, N. Y. Khaireldin, A. F. Fahmy, and A. A. El-Sayed
Vol 000
chlorobenzene), 140.43 (C-14), 145.25 (C-12), 166.03 (C¼O),
92.67 (C¼O). Anal. Calcd. For Molecular Formula
Molecular Formula C21 Cl = C, 65.36%; H, 6.27%;
Cl, 9.19%; N, 10.89%; Found: C, 65.02%; H, 5.98%; Cl,
.99%; N, 10.75%.
-Methyl-4-(4-nitrophenyl)-7-thioxo-1,4,6,8-tetrahydropyra-
zolo[4′,3′:5, 6]pyrano[2,3-d]pyrimidin-5-one (6a). Pale
brown crystals, mp 220–221°C, (EtOH), yield (75%); Ms. m/z:
4-(4-Chlorophenyl)-3-methyl-1,4,6,8-tetrahydropyrazolo[4′,3′:5,6]
pyrano[2,3-d]pyrimidine-5,7-dithione (7b). Pale yellow
crystals, mp 250–252°C, (EtOH), yield (70%); Ms. m/z: 364
1
24 3 2
H N O
+
2
37
+
35
−1
[M , Cl , 19%], 362 [M , Cl , 7%]. IR (KBr) v cm : 3285-
1
8
3185 (NH), 2950 (CH
δ 1.34 (s, 3H, -CH ); 4.56 (s, 1H, -CH), 6.63 (d, 2H, J = 5.4,
aromatic protons), 7.52 (d, 2H, J = 5.4, aromatic protons), 8.01 (s,
1H, NH, D O exchangeable), 11.0 7 (s, 1H, NH, D
exchangeable), 12.10 (s, 1H, NH, D O exchangeable). Anal.
Calcd. For Molecular Formula C15 11ClN OS : C; 49.65%; H,
.03%; Cl, 9.77%; N, 15.44%; Found: C; 49.47%; H, 2.95%; Cl,
.35%; N, 15.21%.
-Methyl-4-phenyl-1,4,6,8,9-pentadropyrazolo[4′,3′:5,6]pyrido
2,3-d]pyrimidine-5,7-dithione (7c). Pale yellow crystals, mp
46–247°C, (EtOH), yield (77%); Ms. m/z: 327 [M , 18%]. IR
3
). H-NMR (270 MHz, DMSO-d , TMS):
6
3
3
2
2
O
+
−1
3
57 [M , 41%]. IR (KBr) v cm : 3280–3195 (NH), 2950 (CH
3
),
2
1
H
4
2
1
690 (C¼O), 1557 (NO ). H-NMR (270 MHz, DMSO-d6,
2
3
9
TMS): δ 1.80 (s, 3H, -CH ), 4.46 (s, 1H, -CH), 7.17-7.50 (m, 4H,
3
aromatic), 10.91 (s, 1H, NH, D
NH, D O exchangeable), 12.10 (s, 1H, NH, D
Anal. Calcd. For Molecular Formula
0.42%; H, 3.10%; N, 19.60%; Found: C, 50.18%; H, 2.98%;
N, 19.23%.
2
O exchangeable), 11.17 (s, 1H,
O exchangeable).
S: C,
3
2
2
[
2
15 11 5 4
C H N O
+
5
−1
1
(
KBr) v cm : 3285–3175 (NH), 2950 (CH ). H-NMR (270
3
MHz, DMSO-d , TMS): δ 1.78 (s, 3H, -CH ), 4.81 (s, 1H, -CH),
4
-(4-Chlorophenyl)-3-methyl-7-thioxo-1,4,6,8-tetrahydropyra-
6
3
7
.18–7.35 (m, 6H, 5 aromatic protons and NH, D O
zolo[4′,3′:5, 6]pyrano[2,3-d]pyrimidin-5-one (6b).
Yellow
2
exchangeable), 10.01 (s, 1H, NH, D
H, NH, exchangeable), 12.23 (s, 1H, NH,
exchangeable). Anal. Calcd. For Molecular Formula C H N S :
2
O exchangeable), 11.95 (s,
crystals, mp 197–198°C, (EtOH), yield (60%); Ms. m/z: 348
+
2
37
+
35
−1
1
2
D O
2
D O
[
3
M , Cl , 14%]; 346 [M , Cl , 6%]. IR (KBr) v cm : 3298–
1
165 (NH), 2950 (CH ), 1690 (C¼O). H-NMR (270 MHz,
15 13 5 2
3
C; 55.02%; H, 4.00%; N, 21.39%; Found: C; 54.95%; H, 3.95%;
N, 21.20%.
DMSO-d
6
, TMS): δ 1.85 (s, 3H, -CH
d, 2H, J = 11.1, aromatic), 7.28 (d, 2H, J = 11.1; aromatic), 9.91
2
s, 1H, NH, D O exchangeable), 10.96 (s, 1H, NH, D O
3
), 4.56 (s, 1H, -CH), 7.15
(
(
4
-(4-Chlorophenyl)-3-methyl-1,4,6,8,9-pentahydropyrazolo
4′,3′:5,6]pyrido[2,3-d]pyrimidine-5,7-dithione (7d).
Yellow crystals, mp 270–271°C, (EtOH), yield (72%); Ms.
2
[
exchangeable), 12.10 (s, 1H, NH, D O exchangeable). Anal.
2
Calcd. For Molecular Formula C H ClN O S: C, 51.95%; H,
1
5
11
4 2
+
2
37
+
35
m/z: 363 [M , Cl ,46%], 361 [M , Cl , 17%]. IR (KBr) v
3
1
.20%; Cl, 10.22%; N, 16.16%; Found: C, 51.61%; H, 2.98%; Cl,
0.01%; N, 15.98%.
−
1
1
cm : 3290–3210 (NH), 2980 (CH
DMSO-d , TMS): δ 1.24 (s, 3H, -CH
.20 (s, 1H, NH, D O exchangeable), 7.49 (d, 2H, J = 8.2,
aromatic), 8.45 (d, 2H, J = 8.1; aromatic), 10.73 (s, 1H,
3
). H-NMR (270 MHz,
6
3
), 4.85 (s, 1H, -CH),
3
-Methyl-4-phenyl-7-thioxo-1,4,6,8,9-pentahydropyrazolo
4′,3′:5,6]pyrido[2,3-d]pyrimidin-5-one (6c). Dark yellow
crystals, mp 205–206°C, (EtOH), yield (62%); Ms. m/z: 311 [M ,
7
2
[
+
−
1
NH, D O exchangeable), 11.15 (s, 1H, NH, D O
6
(
1%]. IR (KBr) v cm : 3290–3210 (NH), 2950 (CH
3
), 1695
2
2
1
exchangeable), 12.15 (s, 1H, NH, D
2
O exchangeable). Anal.
C¼O). H-NMR (270 MHz, DMSO-d
6
, TMS): δ 1.80 (s, 3H, -
Calcd. For Molecular Formula C15
3
3
H
12ClN
.34%; Cl, 9.80%; N, 19.35%; Found: C, 49.42%; H,
.18%; Cl, 9.62%; N, 19.20%.
,7,7-Trimethyl-4-(4-nitrophenyl)-1,4,6,8-tetrahydropyrazolo
4′,3′:5,6] pyrano[2,3-d]pyrimidin-5-one (8a). Brown
crystals, mp 291–292°C, (MeOH), yield (65%); Ms. m/z: 355
5 2
S : C, 49.79%; H,
CH ), 4.65 (s, 1H, -CH), 7.15–7.50 (m, 6H, 5 aromatic protons
3
and NH, D O exchangeable), 8.96 (s, 1H, NH, D O
2
2
2
exchangeable), 10.21 (s, 1H, NH, D O exchangeable), 12.15 (s,
3
1
H, NH, D
2
O exchangeable). Anal. Calcd. For Molecular
OS: C, 57.86%; H, 4.21%; N, 22.49%; Found:
[
Formula C15
H
13
N
5
C, 57.66%; H, 3.98%; N, 22.32%.
-(4-Chlorophenyl)-3-methyl-7-thioxo-1,4,6,8,9-pentahydro-
+
−1
[
1
M , 13%]. IR (KBr) v cm : 3290–3185 (NH), 2950 (CH ),
4
3
1
700 (C¼O), 1557 (NO ). H-NMR (500 MHz, DMSO-d6,
pyrazolo[4′,3′:5, 6]pyrido[2,3-d]pyrimidin-5-one (6d). Yellow
crystals, mp 260–262°C, (EtOH), yield (65%); Ms. m/z: 348
2
TMS): δ 1.81 (s, 3H, -CH ), 2.71 (s, 3H, -CH ), 2.90 (s, 3H, -
3
3
+
2
37
+
35
−1
CH ), 4.51 (s, 1H, -CH), 8.21 (d, 2H, J = 13.10, aromatic), 8.42
[
3
M , Cl , 12%]; 346 [M , Cl , 5%]. IR (KBr) v cm : 3290–
3
1
(
d, 2H, J = 13.10; aromatic), 8.65 (s, 1H, NH, D O
198 (NH), 2950 (CH ), 1692 (C¼O). H-NMR (270 MHz,
2
3
DMSO-d , TMS): δ 2.01 (s, 3H, -CH ), 4.95 (s, 1H, -CH), 7.11
exchangeable), 10.94 (s, 1H, NH, D
2
O exchangeable), 11.22 (s,
O exchangeable). C-NMR (500 MHz, DMSO-d
TMS): δ 17.82 (-CH ), 26.52 (C-5), 32.92 (2-CH ), 55 (C-2),
6
3
13
(
d, 2H, J = 7.1, aromatic), 7.26 (d, 2H, J = 7.1; aromatic), 7.92
1H, NH, D
2
,
6
(
s, 1H, NH, D O exchangeable), 8.73 (s, 1H, NH, D
2
2
O
3
3
exchangeable), 11.15 (s, 1H, NH, D O exchangeable), 12.15
88.65 (C-10), 119 (C-12), 126.21, 129.56, 135.20, 139.52
(aromatic carbons), 142.52 (C-13), 149.52 (C-6), 172.50 (C-11),
179.41 (C¼O). Anal. Calcd. For Molecular Formula
2
(
s, 1H, NH, D O exchangeable). Anal. Calcd. For Molecular
2
Formula C15
N, 20.25%; Found: C, 51.97%; H, 3.33%; Cl, 9.98%; N,
9.98%.
-Methyl-4-(4-nitrophenyl)-1,4,6,8-tetrahydropyrazolo[4',3:'5,6]
5
H12ClN OS: C, 52.10%; H, 3.50%; Cl, 10.25%;
17 17 5 4
C H N O : C, 57.46%; H, 4.82%; N, 19.71%; Found: C,
1
57.11%; H, 4.45%; N, 19.53%.
3
4-(4-Chlorophenyl)-3,7,7-trimethyl-1,4,6,8-tetrahydropyra-
zolo[4′,3′:5,6]pyrano- [2,3-d]pyrimidin-5-one (8b). Yellowish
crystals, mp 180–181°C, (EtOH), yield (60%); Ms. m/z: 346
pyrano[2,3-d]pyrimidine-5,7-dithione (7a). Pale brown crystals,
+
mp 220–221°C, (EtOH), yield (75%); Ms. m/z: 373 [M , 72%]. IR
−1
1
+2
37
+
35
−1
(
KBr) v cm : 3280–3195 (NH), 2950 (CH
3
), 1558 (NO
2
). H-
[M , Cl , 17%], 344 [M , Cl , 5%]. IR (KBr) v cm : 3290–
1
NMR (270 MHz, DMSO-d , TMS): δ 2.02 (s, 3H, -CH
6
3
), 4.26 (s,
3158 (NH), 2900–2850 (CH
MHz, DMSO-d , TMS): δ 1.57 (s, 3H, -CH
2.89 (s, 3H, -CH ), 4.65 (s, 1H, -CH), 7.14 (d, 2H, J = 10.30,
aromatic), 7.39 (d, 2H, J = 10.10; aromatic), 8.94 (s, 1H, NH,
O exchangeable), 10.64 (s, 1H, NH, D O exchangeable), 11.52
(s, 1H, NH, D O exchangeable). Anal. Calcd. For Molecular
3
), 1695 (C¼O). H-NMR (270
1
H, -CH), 7.17–7.50 (m, 4H, aromatic), 10.01 (s, 1H, NH, D O
6
3
), 2.73 (s, 3H, -CH ),
3
2
exchangeable), 11.50 (s, 1H, NH, D O exchangeable), 12.03 (s,
3
2
1
C
4
H, NH, D
2
O exchangeable). Anal. Calcd. For Molecular Formula
: C, 48.25%; H, 2.99%; N, 18.75%; Found: C,
7.98%; H, 2.75%; N, 18. 56%.
15 11 5
H N O
3
S
2
D
2
2
2
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2013
The Utility of Carbon Disulphide and Lawesson's Reagent for
Synthesis of Different Fused Heterocycles for Antimicrobial Evaluation
Formula C H ClN O : C, 59.22%; H, 4.97%; Cl, 10.28%; N,
aromatic), 7.35 (d, 2H, JH-H = 9.95, aromatic), 7.75 (d, 2H, JH-H
=
17
17
4 2
1
6.25%; Found: C, 58.99%; H, 4.76%; Cl, 10.12%; N, 15.98%.
,7,7-Trimethyl-4-phenyl-1,4,6,8,9-pentahydropyrazolo[4′,3′:5,6]
pyrido[2,3-d]pyrimidin-5-one (8c). Pale brown crystals, mp
25–226°C, (toluene), yield (65%); Ms. m/z: 309 [M , 62%]. IR
9.99, aromatic), 8.32 (s, 1H, NH, D O exchangeable), 10.55
2
3
1
3
(s, 1H, NH, D
, TMS): δ +51.56 (s, 1P). Anal. Calcd. For Molecular
Formula C25 25ClN PS : C, 56.65%; H, 4.75%; Cl; 6.69%;
2
O exchangeable). P-NMR (500 MHz, DMSO-
d
6
+
2
H
3
O
2
2
−1
1
(
KBr) v cm : 3299–3215 (NH), 2958 (CH ), 1689 (C¼O). H-
N, 7.93%; P, 5.84%; Found: C, 56.32%; H, 4.51%; Cl; 6.50%;
N, 7.78%; P, 5.66%.
3
NMR (500 MHz, DMSO-d , TMS): δ 2.01 (s, 3H, -CH ), 2.75
6
3
(
(
(
s, 3H, -CH
m, 6H, 5 aromatic protons and NH, D
s, 1H, NH, D O exchangeable), 9.94 (s, 1H, NH, D O
3
), 2.93 (s, 3H, -CH
3
), 4.81 (s, 1H, -CH), 7.03–7.25
2-(4-Methoxyphenyl)-5-(4-(N,N-dimethylamino)phenyl)-6-
dimethyl-1,3,5,8-tetrahydropyrazolo[4′,3′:5,6]pyrano[2,3-d]
[1,3,2]diazaphosphinine-4-thione-2-sulfide (12a). Yellow
crystals, mp 297–298°C, (benzene), yield (50%); Ms. m/z: 497
2
O exchangeable), 8.75
2
2
13
exchangeable), 11.55 (s, 1H, NH, D O exchangeable). C-NMR
2
+
−1
(
(
1
(
500 MHz, DMSO-d
2CH ), 55.25 (C-2), 88.25 (C-10), 109.35 (C-12), 120.21,
25.89, 129.70, 136.52 (aromatic carbons), 140.72 (C-13), 159.32
C-6), 169.59 (C-11), 175.91 (C¼O). Anal. Calcd. For Molecular
Formula C H N O: C, 66.00%; H, 6.19%; N, 22.64%; Found:
6
, TMS): δ 17.52 (-CH
3
), 25.91 (C-5), 35.62
[M , 19%]. IR (KBr) v cm : 3295–3188 (NH), 2950–2890
1
3
(CH
DMSO-d
(s, 3H, -CH ), 3.77 (s, 3H, -OCH ), 4.62 (s, 1H, -CH), 6.73
3
), 1442 (P-C), 821 (P-N), 791 (P¼S). H-NMR (500 MHz,
6
, TMS): δ 1.77 (s, 3H, -CH ), 2.11 (s, 3H, -CH ), 2.28
3
3
3
3
(s, 1H, NH, D O exchangeable), 6.97 (d, 2H, J = 12.8,
17
19
5
2
H-H
C, 65.96%; H, 5.98%; N, 22.28%.
-(4-Chlorophenyl)-3,7,7-trimethyl-1,4,6,8,9-pentahydropy-
aromatic), 7.19 (d, 2H, JH-H = 12.8, aromatic), 7.29 (d, 2H, JH-H
11.30; aromatic), 7.80 (d, 2H, JH-H = 11.30; aromatic), 9.90 (s, 1H,
NH, D O exchangeable), 12.10 (s, 1H, NH, D O exchangeable).
P-NMR (500 MHz, DMSO-d , TMS): δ +49.65 (s, 1P); Anal.
=
4
razolo[4′,3′:5,6]py- rido[2,3-d]pyrimidin-5-one (8d). Greenish
yellow crystals, mp 275–276°C, (Toluene), yield (70%); Ms. m/z:
2
2
31
6
+
2
37
+
35
−1
3
3
45 [M , Cl , 35%], 343 [M , Cl , 14%]. IR (KBr) v cm :
Calcd. For Molecular Formula C H N O PS : C, 55.52%; H,
23 24 5 2 2
1
290–3185 (NH), 2945 (CH
, TMS): δ 1.98 (s, 3H, -CH
.90 (s, 3H, -CH ), 4.81 (s, 1H, -CH), 6.98 (s, 1H, NH, D
exchangeable), 7.78 (d, 2H, J = 7.60, aromatic); 8.39 (d, 2H, J =
3
), 1699 (C¼O). H-NMR (270
4.86%; N, 14.08%; P, 6.22%; Found: C, 55.35%; H, 4.50%; N,
13.98%; P, 5.97%.
2-(4-Methoxyphenyl)-5-(4nitro-phenyl)-6-dimethyl-1,3,5,8-
tetrahydropyrazolo[4′, 3′:5,6]pyrano[2,3-d][1,3,2]diazaphosphinine-
MHz, DMSO-d
6
3
), 2.72 (s, 3H, -CH ),
3
2
3
2
O
7
1
.30; aromatic protons), 9.65 (s, 1H, NH, D O exchangeable),
4-thione-2-sulfide (12b).
(benzene), yield (71%); Ms. m/z: 499 [M , 35%]. IR (KBr) v cm :
White crystals, mp 310–311°C,
2
+
−1
0.32 (s, 1H, NH, D
2
O exchangeable), 11.55 (s, 1H, NH, D
2
O
exchangeable). Anal. Calcd. For Molecular Formula
C H ClN O: C, 59.39%; H, 5.28%; Cl, 10.31%; N, 20.37%;
Found: C, 58.98%; H, 4.99%; Cl, 10.11%; N, 20.12%.
3295–3158 (NH), 2950–2890 (CH ), 1557 (NO ), 1450 (P-C), 825
3
2
1
17
18
5
(P-N), 740 (P¼S). H-NMR (500 MHz, DMSO-d , TMS): δ 2.01
6
(s, 3H, -CH ), 3.76 (s, 3H, -OCH ), 4.82 (s, 1H, -CH), 6.53 (s, 1H,
3
3
General procedure: Synthesis of 11a,b and 12a–d. To a
solution of 1a,b or 5a–d (0.29 or 0.32 g, 0.001 mol), in dry
toluene (15 mL), LR (0.404 g, 0.001 mol) was added. The
reaction mixture was stirred at reflux temperature until no more
of the starting materials were detected by TLC (4–8 h). The
solvent was evaporated under vacuum, and the remaining solids
were crystallized with the proper solvent.
NH, D O exchangeable), 6.97 (d, 2H, J
(d, 2H, JH-H = 12.8, aromatic), 8.29 (d, 2H, JH-H = 13.30,
aromatic); 8.80 (d, 2H, JH-N = 13.30, aromatic), 10.90 (s, 1H, NH,
= 12.8, aromatic), 7.19
2
H-H
31
D O exchangeable), 11.10 (s, 1H, NH, D O exchangeable). P-
2
2
NMR (500 MHz, DMSO-d , TMS): δ +50.23 (s, 1P); Anal.
6
Calcd. For Molecular Formula C H N O PS : C, 50.50%; H,
2
1
18
5
4
2
3.63%; N, 14.02%; P, 6.20%; Found: C, 50.23%; H, 3.21%; N,
13.95%; P, 5.95%.
2
-(4-Methoxyphenyl)-8,8-dimethyl-5-phenyl-4-thioxo-1,3,5,7,9,10-
octahydro[1,3, 2]diazaphosphinino[4,5-b]quinolin-6-one-2-sulfide
11a). Pale yellow crystals, mp 298–299°C, (MeOH), yield
56%); Ms: m/z: 495 [M , 14%]. IR (KBr) v cm : 3295–3185
NH), 2945 (CH
), 1689 (C¼O), 1439 (P-C), 820 (P-N), 785
P¼S). H-NMR (500 MHz, DMSO-d , TMS): δ 0.86 (s, 3H, -
CH ), 1.01 (s, 3H, -CH ), 1.95 (s, 2H, -CH ), 2.29 (s, 2H, -CH ),
2-(4-Methoxyphenyl)-6-methyl-5-phenyl-1,3,5,8,9-pentahydro-
pyrazolo[4′,3′:5,6] pyrido[2,3-d][1,3,2]diazaphosphinine-4-thione-
2-sulfide (12c). White crystals, mp 220–222°C, (benzene), yield
(
(
+
−1
+
−1
(
(
3
(37%); Ms. m/z: 453 [M , 25%]. IR (KBr) v cm : 3295–3210
1
1
6
(NH), 2980–2875 (CH
NMR (500 MHz, DMSO-d , TMS): δ 1.76 (s, 3H, -CH ), 3.76 (s,
3
), 1443 (P-C), 826 (P-N), 782 (P¼S). H-
3
3
2
2
6
3
3
.75 (s, 3H, -OCH ), 4.75 (s, 1H, -CH); 6.98 (s, 1H, NH, D O
3H, -OCH ), 4.57 (s, 1H, -CH), 6.86 (s, 1H, NH, D O
3 2
3
2
exchangeable), 7.03 (d, 2H, J = 9.60, aromatic), 7.39 (d, 2H, J =
.69; aromatic), 7.55–7.98 (m, 5H, aromatic), 10.02 (s, 1H, NH,
exchangeable), 6.97 (d, 2H, JH-H = 12.10, aromatic), 7.13 (d, 2H,
H-H = 12.10, aromatic), 7.25–7.47 (m, 5H, aromatic), 8.95 (s, 1H,
NH, D O exchangeable), 10.06 (s, 1H, NH, D O exchangeable),
9
J
31
D O exchangeable), 10.85 (s, 1H, NH, D O exchangeable). P-
2
2
2
2
13
NMR (500 MHz, DMSO-d , TMS): δ +52.20 (s, 1P) ppm. Anal.
12.10 (s, 1H, NH, D O exchangeable). C-NMR (500 MHz,
6
2
Calcd. For Molecular Formula C25
5
8
H
26
N
3
O
2
PS
.29%; N, 8.48%; P, 6.25%; Found: C, 60.21%; H, 4.98%; N,
.12%; P, 5.99%.
-(4-Methoxyphenyl)-5-(4-chloro-phenyl)-8,8-dimethyl-4-
thioxo-1,3,5,7,9,10-octahydro[1,3,2]diazaphosphinino[4,5-b]
quinolin-6-one-2-sulfide (11b). White crystals, mp 250–
2
: C, 60.59%; H,
6 3 3
DMSO-d , TMS): δ 10.42 (-CH ), 31.78 (C-5), 100.54 (OCH ),
104.81 (C-10), 111.87 (C-12), 112.30, 118.59, 131.18, 154.00
(p-methoxyphenyl), 127.90, 128.54, 132.30, 137.17 (Phenyl
carbons), 148.63 (C-6), 155.43 (C-13), 161.16 (C-11), 192.32
2
31
(C-4). P-NMR (500 MHz, DMSO-d , TMS): δ +48.83 (s, 1P);
6
20 5 2
Anal. Calcd. For Molecular Formula C21H N OPS : C, 55.61%;
H, 4.44%; N, 15.44%; P, 6.83%; Found: C, 55.33%; H, 4.21%;
N, 14.99%; P, 6.59%.
5-(4-Chlorophenyl)-2-(4-methoxyphenyl)-6-methyl-1,3,5,8,9-
pentahydropyrazolo [4′,3′:5,6]pyrido[2,3-d][1,3,2]diazaphosphinine-
+
2
37
2
5
51°C, (toluene), yield (66%); Ms. m/z: 532 [M , Cl , 6%],
30 [M , Cl , 4%]. IR (KBr) v cm : 3290–3195 (NH), 2945
+
35
−1
1
(
CH ), 1696 (C¼O), 1441 (P-C), 825 (P-N), 805 (P¼S). H-
3
NMR (500 MHz, DMSO-d , TMS): δ 0.86 (s, 3H, -CH ), 1.01
6
3
(
-
6
s, 3H, -CH
OCH
.97 (d, 2H, JH-H = 12.20, aromatic), 7.13 (d, 2H, JH-H = 12.10;
3
), 1.95 (s, 2H, -CH
), 4.65 (s, 1H, CH), 6.78 (s, 1H, NH, D O exchangeable),
2
2
), 2.29 (s, 2H, -CH
2
), 3.75 (s, 3H,
4-thione-2-sulfide (12d).
White crystals, mp 270–272°C,
+2
37
+
3
(toluene), yield (65%); Ms. m/z: 489 [M , Cl , 10%], 487 [M ,
35
−1
3
Cl , 4%]. IR (KBr) v cm : 3290–3201 (NH), 2955–2892 (CH ),
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

N. R. Mohamed, N. Y. Khaireldin, A. F. Fahmy, and A. A. El-Sayed
Vol 000
1
1
445 (P-C), 830 (P-N), 800 (P¼S). H-NMR (500 MHz, DMSO-d ,
the zone diameters were measured with slipping calipers of the
National Committee for Clinical Laboratory Standards.
Agar-based methods such as test and disk diffusion can be
good alternatives because they are simpler and faster than
both-based methods [25].
6
TMS): δ 1.79 (s, 3H, CH ), 3.77 (s, 3H, -OCH ), 4.60 (s, 1H, CH),
3
3
6
.86 (s, 1H, NH, D
aromatic), 7.30 (d, 2H, JH-H = 11.9, aromatic), 7.58 (d, 2H, JH-H
.30, aromatic), 8.80 (d, 2H, JH-H = 9.30; aromatic), 9.03 (s, 1H,
NH, D O exchangeable), 10.90 (s, 1H, NH, D O exchangeable),
2
O exchangeable), 6.97 (d, 2H, JH-H = 11.90,
=
9
2
2
31
1
1.10 (s, 1H, NH, D O exchangeable). P-NMR (500 MHz,
2
DMSO-d
6
, TMS): δ +50.23 (s, 1P); Anal. Calcd. For Molecular
19ClN OPS : C, 51.69%; H, 3.92%; Cl, 7.27%; N,
4.35%; P, 6.35%; Found: C, 51.45%; H, 3.52%; Cl, 7.03%; N,
4.02%; P, 5.98%.
Antimicrobial activity. Antimicrobial activity of the tested
samples was determined using a modified Kirby-Bauer disc
diffusion method [22]. Briefly, 100 μL of the test bacteria/fungi
were grown in 10 mL of fresh media until they reached a count
of ∼108 cells/mL for bacteria or 105 cells/mL for fungi. A total
of 100 μL of microbial suspension was spread onto agar plates
Formula C21
H
5
2
REFERENCES AND NOTES
1
1
[1] Valverde, M. G.; Torroba, T.; Sulfur-Nitrogen Heterocycl Mol
2005, 10, 318.
[
2] Balaba, A. T. N.; Oniciu, D. C.; Katritzky, A.R. Chem Rev
2
004, 104, 2777.
[
[
[
3] Abdou, M. I.; Saleh, A. M.; Zohdi, H. F. Molecules 2004, 9, 109.
4] Timothy, W.; Joseph, L. Curr Pharm Des 2007, 13, 1.
5] Alhiari, M. Y.; Abu-Dahab, R.; El-Abadekah, M. M.
Molecules 2008, 13, 11.
corresponding to the broth in which they were maintained.
Isolated colonies of each organism that might be playing a
[6] Oleg, A. P. Arkivoc (i) 2009, ISSN 1551–7012, 129.
[
7] Stanetty, P.; Turner, M.; Mihovilovic, M. Molecules 2005, 10, 367.
[8] Ram, V. J.; Goel, A.; Sarkhel, S.; Maulik, P. R. Bioorg Med
Chem 2002, 10, 1275.
9] Hino, T.; Nakagawa, M. J Heterocycl Chem 2009, 31, 625.
10] Polshettiwar, V.; Varma, S.-R. Pure Appl Chem 2008, 80, 777.
pathogenic role should be selected from primary agar plates and
tested for susceptibility by disc diffusion method. Of the many
media available, NCCLS recommends Mueller-Hinton agar due
to: it results in good batch-to-batch reproducibility. Disc diffusion
method for filamentous fungi tested by using approved standard
method (M38-A) [23] for evaluating the susceptibilities of fila-
mentous fungi to antifungal agents. Disc diffusion method for
yeasts developed by using approved standard method (M44-P)
[
[
[
[
[
11] Zayed, E. S. Phosphorous Sulfur Silicon Relat Elem 2007, 182, 2193.
12] Tominaga, Y. S. J Heterocycl Chem 2009, 26, 1167.
13] Elkateb, A.; Abd-AlRahman, N. M. Phosphorous Sulfur
Silicon Relat Elem 2006, 181, 249.
[14] KhirEldin, N. Y.; Gad, W. A.; Ali, Y. M.; Mohamed, N. R.
Egypt J Chem 2007, 50, 531.
[
24]. Plates inoculated with filamentous fungi as Aspergillus
[
15] Mohamed, N. R.; Elmegeed, G. A.; Abd-ElMalek, H. A.;
Younis, M. Steroids 2005, 70, 131.
16] Mohamed, N. R.; Elmegeed, G.; Younis, M. Phosphorous Sul-
fur Silicon Relat Elem 2003, 178, 2003.
17] Mohamed, N. R.; Elsaidi, M. M. T.; Abdallah, T.; Nada, A. A.;
Phosphorous Sulfur Silicon Relat Elem 2004, 179, 2387.
[18] Wang, X. S.; Zhang, M. M.; Jiang, H.; Yao, C. S.; Tu, S. J.
Tetrahedron 2007, 63, 4439.
Flavus at 25°C for 48 h; Gram positive bacteria as Staphylococcus
Aureus, Bacillus Subtilis; Gram negative bacteria as Escherichia
Coli, Pseudomonas Aeuroginosa they were incubated at 35–37°C
for 24–28 h and yeast as Candida Albicans incubated at 30°C for
[
[
2
4–28 h then the diameters of the inhibition zones were measured
in millimeters [22].
Standers discs of Tetracycline (Antibacterial agent),
Amphotericin B (Antifungal agent) served as positive controls
for antimicrobial activity but filter discs impregnated with 10 μL
of solvent (distilled water, chloroform, DMSO) were used as a
negative control. The agar used is Meuller-Hinton agar in the
plate is a factor to be considered in the disc diffusion method.
This method is well documented and standard zones of inhibition
have been determined for susceptible and resistant values.
Blank paper disks (Schleicher & Schuell, Spain) withy a
diameter of 8.0 mm were impregnated 10 μL of tested concen-
tration of the stock solutions. When a filter paper disc impreg-
nated with a tested chemical is placed on agar the chemical in
the agar only around the disc. The solubility of the chemical
infiltration around the disc. If an organism is placed on the agar,
it will not grow in the area around the disc if it is susceptible to
the chemical. This area of on growth around the disc is known as
a “Zone of inhibition” or “Clear zone.” For the disc diffusion,
[19] Meryers, A. I.; Ford, M. E. J Org Chem 1976, 41, 1735.
[20] Mohamed, N. R.; Khaireldin, N. Y.; Fahmy, A. F.; El-Sayed,
A. A. Pharm Chem 2010, 2, 400.
[
[
21] Deng, S.-L.; Chen, R.-Y. Monatsh Chem 2004, 135, 1113.
22] Bauer, A. W.; Kirby, W. M.; Sherris, J. C..; Turk, M. Am J
Clin Pathol 1966, 45, 493.
[23] National Committee for Clinical Laboratory Standards.
Reference Method for Broth Dilution Antifungal Susceptibility testing
of Conidium-forming Filamentous Fungi: Proposed Standard M38-A.
NCCLS, PA, USA. National Committee for Clinical Laboratory Stan-
dards, 2002.
[24] National Committee for Clinical Laboratory Standards.
Method for Antifungal Disk Diffusion Susceptibility testing of Yeast: Pro-
posed Guideline M44-P. NCCLS, Wayne, PA, USA. National Committee
for Clinical Laboratory Standards, 2003.
[25] Matar, M. J.; Ostrosky-Zeichner, L.; Paetznick, V. L.;
Rodriguez, J. R.; Chen E.; Rex, J. H. Antimicrob Agents Chemother
2003, 47, 1647.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet