Site- and regioselectivity of the reaction of hydrazonoyl chlorides with perimidine ketene aminal. Antimicrobial evaluation of the products

Article abstract of DOI:10.1002/jhet.1985

Reaction of hydrazonoyl chlorides with perimidine ketene aminal derivative in dioxane in the presence of triethylamine afforded either pyrrolo[1,2-a]perimidines or pyrazolyl perimidines depending on the type of hydrazonoyl chloride used. The reaction was

Full text of DOI:10.1002/jhet.1985

Month 2014 Site- and Regioselectivity of the Reaction of Hydrazonoyl Chlorides with
Perimidine Ketene Aminal. Antimicrobial Evaluation of the Products
*
Thoraya A. Farghaly and Huda K. Mahmoud
Chemistry Department, Faculty of Science, University of Cairo, Giza 12613, Egypt
*
E-mail: thoraya-f@hotmail.com
Received July 2, 2012
DOI 10.1002/jhet.1985
Published online 00 Month 2014 in Wiley Online Library (wileyonlinelibrary.com).
Reaction of hydrazonoyl chlorides with perimidine ketene aminal derivative in dioxane in the presence of
triethylamine afforded either pyrrolo[1,2-a]perimidines or pyrazolyl perimidines depending on the type of
hydrazonoyl chloride used. The reaction was found to be site- and regioselective according to the suggested
mechanism. The structure of the newly synthesized compounds was established on the basis of spectral data
and elemental analyses. In addition, the antimicrobial activity of the newly synthesized compounds was
evaluated, and the results showed moderate activity of all compounds against the bacterial species.
J. Heterocyclic Chem., 00, 00 (2014).
INTRODUCTION
1,8-diamino-naphthalene [22,23]. Recently [24], compound 3
was prepared by the reaction of 1,8-diamino-naphthalene 1
with ethyl benzoylacetate 2 under reflux for 6 h in xylene in
the presence of P-toluenesulfonic acid (PTSA catalytic
amount) in good yields (56–73%). In this work, we report here
a new approach for the preparation of 3. Thus, fusion of 1
with ethyl benzoylacetate 2 without solvent for 15 min
afforded compound 3 in very good yield (90%). The structure
assignment of 3 was confirmed using X-ray crystallographic
analysis [25] (Fig. 2 and Table 1).
Hydrazonoyl halides have been widely employed in the
synthesis of heterocyclic derivatives [1–3]. Reaction of
heterocyclic ketene aminal I, which are the cyclic enediamine
analogues bearing secondary amino group(s), with nitrilimine
II led to the formation of pyrazole III [4–6] (Fig. 1). Such
results were considered to indicate that reaction of nitrilimine
with ketene aminal is 1,3-cycloaddition condensation and
stepwise reaction. In addition, perimidines and their deriva-
tives have found a variety of applications ranging from dye-
stuffs [7–11] to fluorescent molecular sensors [12]. Their
biological activity as antimicrobial and antifungal agents,
DNA-binding properties, and antitumor activity of various
perimidines have been studied [8–10,13–15]. Also, they are
found to have the thermochromic properties and EPR spectra
of perimidine (diazaphenalenyl) radicals [12,14–16].
On the basis of the above findings and in continuation
of our recent work in the utility of hydrazonoyl halides in
synthesis of heterocyclic systems with remarkable biological
importance [17–20], we report herein a study to investigate
the site- and regioselectivity of the reaction of hydrazonoyl
chlorides with perimidine ketene aminal. Also, antimicrobial
activity of the newly synthesized compounds was evaluated.
Reaction of compound
3 with N-aryl 2-oxo-2-
phenylaminoethane-hydrazonoyl chlorides 5a–h in diox-
ane solution under reflux in the presence of triethylamine
as a basic catalyst can be afforded theoretically one or more
of seven products 6–12, but this reaction afforded only a
single product as evidenced by TLC (Scheme 2). It
afforded products 7a–h. The structure of compounds 7
was established on the basis of spectral data and elemental
analysis. The mass spectra of all derivatives 7a–h revealed
in each case a molecular ion peak corresponding to elimi-
nation of HCl and water molecules. 1H NMR spectra
of compounds 7 indicated the presence of two singlet
signals at d 10.66–10.43 and 10.94–10.75 ppm for the
two NH groups. In addition, the IR spectra revealed the
presence of absorption bands at 3409–3188, 3200–3128,
and 1659–1627 cmÀ1 characteristic for two NH and amidic
CO groups. On the basis of the above data, products 8, 9,
11, and 12 were discarded. This result indicated that
reaction of ketene aminal 3 with N-aryl 2-oxo-2-
phenylaminoethane-hydrazonoyl chlorides 5a–h proceeded
via intermediate 6 rather than 10.
RESULTS AND DISCUSSION
Barchet et al. [21] have reported that reaction of
1,8-diamino-naphthalene 1 with ethyl benzoylacetate
2 afforded diazocine derivative 4 (Scheme 1). How-
ever, 2-(benzoylmethylene)-1,2-dihydro-perimidine (3)
has been previously reported from the reaction of the
corresponding diketone or 3,3-dichloro-acryloyl chloride with
On the other hand, when the reaction of compound 3
was repeated with ethyl (N-arylhydrazono)-chloroacetates
© 2014 HeteroCorporation

T. A. Farghaly and H. K. Mahmoud
Vol 000
8a–h revealed in each case a molecular ion peak correspond-
ing to the elimination of HCl and ethanol molecules. The spec-
tra also displayed significant peaks corresponding to M-105
and 105. These ions might originate via expulsion of (PhCO)
from the molecular ion. In addition, IR spectra of 8a–h
revealed four bands at 3288–3158, 3267–3158, 1693–1685,
and 1647 cmÀ1 for two NH and two carbonyl groups.
As shown in Chart 1, compounds 8 can have one of the
four tautomeric forms, the hydrazone tautomeric form 8A
and 8B or azo-tautomeric form 8C and 8D. To distinguish
between the four forms, we record the electronic absorp-
tion spectral data of compounds 8a–h (Table 2). As shown,
each of the compounds 8 in dioxane exhibits two charac-
teristic absorption bands in the regions lmax 485–465 and
342–336 nm. Such an absorption pattern is similar to that
of typical azo chromophore [20]. Furthermore, the spec-
trum of 8c, taken as a typical example of the series studied,
in solvents of different polarities showed little, if any, shift
(Table 2). On the basis of these data, it was suggested that
compounds 8 exist in the solution state in one tautomeric
form 8C or 8D and excludes the hydrazone tautomeric
form 8A and 8B (Scheme 3).
Figure 1. Reaction of heterocyclic ketene aminal I with nitrilimine II led
to the formation of pyrazole III.
Scheme 1. Synthesis of compound 3.
To distinguish between the two azo forms 8C and 8D, the
1H NMR spectra of the products exhibit one NH and
one OH (D2O exchangeable) singlet signals near d 12 ppm
and luck the doublet signal at d 5–6 ppm for C─H of
tautomer 8C [26]. Also, the structure of compounds
8 was further evidenced by alternative synthesis via reac-
tion of ketene aminal 3 and methyl (N-(4-chlorophenyl)
hydrazono)chloroacetate Ae under the same reaction
conditions to afford a product that proved to be identical
in all respects (mp, mixed mp, and IR) with compound
8e (Scheme 3). From all the above data, we can conclude
that the product found in the azo tautomeric form 8D in
the solution, while in the solid state (from IR), is found
in the hydrazone form 8A. The foregoing results suggest
that the reactions of ketene aminal 3 with hydrazonoyl
chloride 5 or 13 under reflux in the presence of basic
catalyst leading to the formation of 7 or 8, respectively,
are site- and regioselective and proceed via the intermedi-
ates 6 and 14 depicted in Schemes 2 and 3. Also the differ-
ence in the products can be attributed to the fact that the
perimidine derivatives are used as nucleophilic catalyst
for ester hydrolysis [27].
Figure 2. Crystal structure of compound 3.
Table 1
Selected bond lengths and bond angles in the ORTEP figure of compound
3 in the crystal. The crystallographic numbering does not reflect
systematic numbering.
Bond length (Å)
Bond length (Å)
Angle (o)
Antimicrobial activities. In vitro antimicrobial screening of
compounds 7a,c,e,f and 8a–h prepared in this study was
carried out using two bacteria species, including Gram
negative bacteria, Escherichia coli (RCMB 000103) (EC),
and Gram positive bacteria, Staphylococcus aureus (RCMB
000106) (SA). Two fungal strains include Aspergillus
fumigatus (RCMB 002003) (AF) and Candida albicans
(RCMB 005002) (CA). The results of the investigation
showed that compounds 7a,c,e,f, 8a–c, and 8e–h displayed
significant activities against E. coli and S. aureus, whereas
N5—C16, 1.345
N3—C16, 1.362
C10—C16, 1.399 N3—C12, 1.397 C12—C8—C37, 120.2
C7—C10, 1.393
C8—C12, 1.414
O1—C7, 1.277
C7—C43, 1.494 C16—N5—C37, 124.2
C12—N3—C16, 123.97
C8—C37, 1.409 C7—C10—C16, 122.7
C8—C9, 1.411 O1—C7—C10, 122.6
(13a–h) under the same reaction conditions, it afforded prod-
ucts 8a–h (Scheme 3). The mass spectra of all derivatives
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2014
Site- and Regioselectivity of the Reaction of Hydrazonoyl Chlorides with Perimidine
Ketene Aminal. Antimicrobial Evaluation of the Products
Scheme 2. Synthesis of compounds 7a–h.
Scheme 3. Synthesis of compounds 8a–h.
Chart 1. The tautomeric structures of compound 8.
Table 2
UV Spectra of compounds 8 in dioxane.
Compound no.
lmax (log e)
8a
8b
8ca
8d
8e
8f
472 (3.95), 341 (3.73)
471 (4.17), 342 (3.99)
471 (4.02), 342 (4.0)
465 (3.82), 341 (3.72)
471 (4.31), 341 (4.03)
466 (3.97), 341 (3.89)
467(4.08), 336 (3.99)
485 (3.89), 342 (3.84)
8g
8h
aSolvent lmax (log e): acetone 463 (4.25), 339 (4.06); chloroform
463 (4.21), 316 (4.05); cyclohexane 468 (4.65), 339 (4.45);
dimethylformamide 463 (4.35), 397 (4.54); ethanol 464 (4.01),
339 (3.98).
all tested compounds have no activity against Aspergillus
flavus and C. albicans (Table 3).
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

T. A. Farghaly and H. K. Mahmoud
Vol 000
Dx = 2.562 mg mÀ3 reflection 12,534 measured,
Figure 1 illustrates the structure as determined [25].
θ
max = 27.51ꢀ.
Table 3
Antimicrobial activity of the newly synthesized compounds 7 and 8.
Synthesis of 2-(benzoylmethylene)-1,2-dihydro-perimidine
(3). A mixture of 1,8-diaminonaphthalene (1) (1.58, 10 mmol) and
ethyl benzoylacetate 2 (2.9g, 15 mmol) was heated to reflux for
15min, then cooled. The solid formed was collected by filtration
and washed with ethanol, then crystallized from dioxane/ethanol
mixture to give compound 3 as a yellowish green crystal, yield
90%, mp 248–250ꢀC [22], (EtOH/Dioxane), IR ύ 3460, 3352
Microorganism/IZD (mm/mg sample)a
Cpd. no.
EC (GÀ)
SA (G+)
AF
CA
Control: DMSO
0.0
13
10
12
11
11
10
11
0.0
10
10
12
10
32
—
0.0
13
11
13
12
13
10
14
0.0
0.0
12
11
11
29
—
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
—
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
—
7a
7c
7e
1
(2NH), 1654 (CO) cmÀ1; H NMR (DMSO-d6) 5.54 (s, 1H, CH),
7f
6.58–7.82 (m, 11H, Ar─H), 10.70 (s, 1H, NH), 13.77 (s, 1H, NH);
ms m/z (%) 286 (M+, 100), 285 (66), 222 (34), 209 (20), 190 (16),
180 (18), 154 (13), 128 (26), 115 (23), 105 (97), 104 (31), 95 (30),
91 (11), 77 (79). Anal. Calcd for C19H14N2O (286.31): C, 79.7; H,
8a
8b
8c
8d
8e
4.92; N, 9.78. Found: C, 79.54; H, 4.86; N, 9.58%.
Reaction of compound 3 with hydrazonoyl chlorides 5 or
13 or Ae. To a mixture of 3 (1.43g, 5 mmol) and the appropriate
hydrazonoyl chloride 5a–h or 13a–h or Ae (5 mmol of each) in
dioxane (30mL) was added triethylamine (0.7 mL), and the
mixture was heated to reflux for 5 h, then cooled. The solid
produced was collected by filtration and crystallized from the
appropriate solvent to give the corresponding compounds 7a–h or
8a–h, respectively. The products 7a–h and 8a–h together with
their physical constants are listed below.
8f
8g
8h
Teb
Amb
18
19
aIZD, inhibition zone diameter,
bTe = tetracycline as standard antibacterial agent and Am = amphotericin B
as standard antifungal agent.
2-[1-(4-Methylphenyl)-3-phenylcarbamoyl-5-phenyl-pyrazol-
4-yl]-perimidine (7a). Pale green, yield 1.2, 95%, mp 228–230ꢀC
1
(EtOH/Dioxane), IR ύ: 3260, 3128 (2NH), 1655 (CO) cmÀ1; H
CONCLUSIONS
NMR (DMSO-d6) 2.18 (s, 3H, CH3), 6.64 (d, J = 8 Hz, 2H,
Ar─H), 6.95–7.46 (m, 16H, Ar─H), 7.81 (d, J = 8 Hz, 2H,
Ar─H), 10.66 (s, 1H, NH), 10.75 (s, 1H, NH). 1H NMR
(DMSO-d6): 20.74, 101.79, 113.34, 113.34, 117.39, 117.55,
119.01, 120.38, 121.57, 122.77, 123.73, 126.17, 127.82,
127.94, 128.38, 128.50, 128.76, 129.16, 129.28, 129.63,
135.09, 138.42, 138.91, 143.47, 144.62, 145.20, 148.72,
159.30. ms m/z (%) 519 (M+, 16), 258 (18), 164 (20), 154 (15),
143 (16), 118 (35), 105 (100), 77 (55). Anal. Calcd for
C34H25N5O (519.57): C, 78.59; H, 4.84; N, 13.48. Found: C,
78.35; H, 4.64; N, 13.18%.
From all the above data, it was concluded that reaction
of hydrazonoyl chlorides with perimidine ketene aminal
proceeds site and regioselective reactions depending on
the structure of the hydrazonoyl chlorides used. The newly
synthesized perimidines showed moderate activity against
the bacteria species used.
EXPERIMENTAL
2-[1-(3-Methylphenyl)-3-phenylcarbamoyl-5-phenyl-pyrazol-
4-yl]-perimidine (7b). Brown solid, yield 1.25, 96%, mp
200–202ꢀC (EtOH/Dioxane), IR ύ: 3379, 3186 (2NH), 1632
All melting points were determined on an electrothermal
Gallenkamp apparatus and are uncorrected. Solvents were generally
distilled and dried prior their use. The IR spectra were measured on
a Pye-Unicam SP300 (Pye Unicam Ltd., Cambridge, UK) instru-
ment in potassium bromide discs. The 1H NMR spectra were
recorded on a Varian Mercury VXR-300 spectrometer (300 MHz
for 1H NMR), and the chemical shifts were related to that of the sol-
vent DMSO-d6. The mass spectra were recorded on a GCMS-
Q1000-EX Shimadzu and GCMS 5988-A HP spectrometers; the
ionizing voltage was 70 eV. Elemental analyses and antimicrobial
activity of the products were carried out at the Microanalytical Cen-
ter of Cairo University, Giza, Egypt. Hydrazonoyl halides 5 and 13
[28] were prepared following literature procedures.
Crystallographic analysis. The crystals were mounted on a
glass fiber. All measurements were performed on an ENRAF
NONIUS FR 590. The data were collected at a temperature of
25ꢀC using the o scanning technique to a maximum of a 20 of
22.98ꢀ. The structure was solved by direct method using SIR 92
and refined by full-matrix least squares. Nonhydrogen atoms
were refined anisotropically. Hydrogen atoms were located
geometrically and were refined isotropically.
1
(CO) cmÀ1; H NMR (DMSO-d6), 2.27 (s, 3H, CH3), 6.17–7.84
(m, 20H, Ar─H), 10.43 (s, 1H, NH), 10.86 (s, 1H, NH). ms m/z
(%) 519 (M+, 51), 426 (90), 425 (100), 191 (18), 140 (16), 119
(51), 105 (6), 93 (51), 91 (88), 77 (24). Anal. Calcd for
C34H25N5O (519.57): C, 78.59; H, 4.84; N, 13.48. Found: C,
78.45; H, 4.66; N, 13.30%.
2-[1,5-Diphenyl-3-phenylcarbamoyl-pyrazol-4-yl]-perimidine
(7c). Yellow solid, yield 1.2, 95%, mp 228–230ꢀC (EtOH/Dioxane),
1
IR ύ: 3409, 3144 (2NH), 1633 (CO) cmÀ1; H NMR (DMSO-d6)
7.02–7.82 (m, 21H, Ar─H), 10.45 (s, 1H, NH), 10.85 (s, 1H, NH).
ms m/z (%) 505 (M+, 4), 412 (4), 308 (2), 190 (7), 178 (8), 166
(12), 154 (10), 139 (8), 126 (10), 105 (12), 91 (38), 77 (25), 69
(56). Anal. Calcd for C33H23N5O (505.54): C, 78.39; H, 4.58; N,
13.85. Found: C, 78.16; H, 4.43; N, 13.68%.
2-[1-(3-Chlorophenyl)-3-phenylcarbamoyl-5-phenyl-pyrazol-4-
yl]-perimidine (7d). Yellow solid, yield 1.32, 98%, mp 224–226ꢀC
1
(EtOH/Dioxane), IR ύ: 3311, 3155 (2NH), 1659 (CO) cmÀ1; H
NMR (DMSO-d6) 6.18–7.82 (m, 20H, Ar─H), 10.46 (s, 1H, NH),
10.88 (s, 1H, NH). ms m/z (%) 541 (M+ + 2, 27), 540 (M+ +1, 14),
539 (M+, 34), 191 (10), 178 (12), 140 (17), 127 (28), 111 (25),
105 (38), 93 (100), 77 (83). Anal. Calcd for C33H22N5ClN5O
Crystal data. For compound 3: C19H14N2O, M= 286.31, triclinic,
a= 14.2459(3), b= 15.1254(5), c= 15.9448(8) Ǻ, v= 2969.9(2),
a = 90.00ꢀ, b = 120.183(18)ꢀ, g =90.00ꢀ, space group: Z=8,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2014
Site- and Regioselectivity of the Reaction of Hydrazonoyl Chlorides with Perimidine
Ketene Aminal. Antimicrobial Evaluation of the Products
(539.98): C, 73.39; H, 4.10; N, 12.97. Found: C, 73.20; H, 4.02;
8-Benzoyl-9-(3-chlorophenylhydrazono)-7H-pyrrolo[1,2-a]
perimidin-10-one (8d). Orange solid, yield 1.12, 96%, mp
278–280ꢀC, (EtOH/Dioxane), IR ύ: 3437, 3210 (2NH), 1693,
N, 12.74%.
2-[1-(4-Chlorophenyl)-3-phenylcarbamoyl-5-phenyl-pyrazol-4-
yl]-perimidine (7e). Yellow solid, yield 1.31, 97%, mp 252–254ꢀC
1
1647 (2CO) cmÀ1, H NMR (DMSO-d6) 6.80–8.46 (m, 15H, Ar
1
(EtOH/Dioxane), IR ύ: 3391, 3200 (2NH), 1644 (CO) cmÀ1; H
H), 11.9 (s, 1H, NH), 12.0 (s, 1H, OH). ms m/z (%), 466 (M+ + 2,
22), 465 (M+ + 1, 23), 464 (M+, 59), 463 (67), 446 (28), 356 (19),
339 (13), 310 (42), 233 (15), 139 (23), 127 (34), 105 (94), 77
(100). Anal. Calcd for C27H17ClN4O2 (464.88): C, 69.75; H, 3.68;
NMR (DMSO-d6) 6.18–7.42 (m, 16H, Ar─H), 7.53 (d, J =8 Hz,
2H, Ar─H), 7.79 (d, J = 8 Hz, 2H, Ar─H), 10.44 (s, 1H, NH),
10.83 (s, 1H, NH). ms m/z (%) 541 (M+ + 2, 33), 540 (M+ + 1, 33),
539 (M+, 78), 446 (100), 178 (35), 140 (30), 126 (10), 111 (50),
92 (23), 77 (88). Anal. Calcd for C33H22ClN5O (539.98): C,
N, 12.05. Found: C, 69.61; H, 3.54; N, 12.17%.
8-Benzoyl-9-(4-chlorophenylhydrazono)-7H-pyrrolo[1,2-a]
73.39; H, 4.10; N, 12.97. Found: C, 73.16; H, 4.05; N, 12.88%.
perimidin-10-one (8e). Orange solid, yield 1.16, 94.6%,
mp 289–300ꢀC, (EtOH/Dioxane), IR ύ: 3401, 3217 (2NH),
2-[1-(4-Bromophenyl)-3-phenylcarbamoyl-5-phenyl-pyrazol-
1690, 1647 (2CO) cmÀ1
,
1H NMR (DMSO-d6) 6.84
4-yl]-perimidine (7f). Pale orange solid, yield 1.35, 92%,
mp 238–240ꢀC (EtOH/Dioxane), IR ύ: 3375, 3190 (2NH), 1642
(d, J = 8 Hz, 2H, Ar H), 7.13 (d, J = 8 Hz, 2H, Ar H), 7.48–
8.45 (m, 11H, Ar H), 11.84 (s, 1H, NH) 12.04 (s, 1H, OH),
ms m/z (%), 466 (M+ + 2, 21), 465 (M+ + 1, 19), 464 (M+, 48),
359 (16), 311 (14), 310 (28), 255 (11), 191 (12), 127 (28),
111 (29), 105 (89), 77 (100). Anal. Calcd for C27H17ClN4O2
(464.88): C, 69.75; H, 3.68; N, 12.05. Found: C, 69.58; H,
1
(CO) cmÀ1; H NMR (DMSO-d6) 6.16–7.39 (m, 16H, Ar─H),
7.67 (d, J = 8 Hz, 2H, Ar─H), 7.79 (d, J = 8 Hz, 2H, Ar─H),
10.47 (s, 1H, NH), 10.85 (s, 1H, NH). ms m/z (%) 586 (M+ + 2,
23), 585 (M+ + 1, 68), 584 (M+, 70), 490 (98), 489 (100), 260
(23), 190 (16), 178 (19), 166 (15), 156 (15), 154 (17), 140 (27),
126 (7), 77 (44). Anal. Calcd for C33H22BrN5O (584.43): C,
67.81; H, 3.79; N, 11.98. Found: C, 67.61; H, 3.67; N, 11.82%.
2-[1-(3-Nitrophenyl)-3-phenylcarbamoyl-5-phenyl-pyrazol-4-
yl]-perimidine (7g). Dark brown solid, yield 1.36, 99%, mp
130–132ꢀC (EtOH/Dioxane), IR ύ: 3188, 3051 (2NH), 1627
3.49; N, 11.95%.
8-Benzoyl-9-(4-bromophenylhydrazono)-7H-pyrrolo[1,2-a]
perimidin-10-one (8f). Orange solid, yield 1.2, 94%, mp
292–294ꢀC (EtOH/Dioxane), IR ύ: 3432, 3267 (2NH), 1689,
1
1647 (2CO) cmÀ1, H NMR (DMSO-d6), 6.79 (d, J = 9 Hz, 2H,
(CO) cmÀ1 1H NMR (DMSO-d6) 7.11–8.34 (m, 20H, Ar─H),
;
Ar H), 7.27 (d, J = 9 Hz, 2H, Ar H), 7.50–8.48 (m, 11H, Ar H),
11.86 (s, 1H, NH), 12.04 (s, 1H, OH), ms m/z (%) 510 (M+ + 1,
19), 509 (M+, 28), 491 (24), 339 (30), 268 (10), 191 (11), 155
(15), 140 (16), 126 (9), 105 (100), 77 (88). Anal. Calcd for
C27H17BrN4O2 (509.33): C, 63.66; H, 3.36; N, 11.0. Found: C,
10.59 (s, 1H, NH), 10.94 (s, 1H, NH). ms m/z (%) 550 (M+, 16),
520 (4), 191 (14), 167 (22), 140 (13), 126 (16), 123 (43), 105
(61), 93 (80), 77 (100). Anal. Calcd for C33H22N6O3 (550.57): C,
71.99; H, 4.03; N, 15.26. Found: C, 71.80; H, 4.15; N, 15.13%.
2-[1-(4-Nitrophenyl)-3-phenylcarbamoyl-5-phenyl-pyrazol-4-
63.54; H, 3.21; N, 11.24%.
yl]-perimidine (7h). Brown solid, yield 1.13, 82%, mp 140–142ꢀC
(EtOH/Dioxane), IR ύ: 3387 (br, 2 NH), 1632 (CO) cmÀ1. ms m/z
(%) 550 (M+, 3), 458 (1), 430 (2), 399 (1), 166 (7), 154 (8), 140 (8),
126 (7), 105 (100), 77 (77). Anal. Calcd for C33H22N6O3 (550.57):
8-Benzoyl-7H-9-(3-nitrophenylhydrazono)-pyrrolo[1,2-a]
perimidin-10-one (8g). Red solid, yield 1.13, 96%, mp 284–286ꢀC,
( EtOH/Dioxane), IR ύ: 3262, 3158 (2NH), 1685, 1647 (2CO)
cmÀ1 1H NMR (DMSO-d6) 7.25–8.46 (m, 15H, Ar─H), 11.9
,
(s, 1H, NH), 12.21 (s, 1H, OH). ms m/z (%), 475 (M+, 70),
457 (11), 445 (6), 339 (13), 310 (48), 233 (14), 166 (14),
140 (12), 127 (11), 105 (100), 77 (100). Anal. Calcd for
C27H17N5O4 (475.44): C, 68.2; H, 3.6; N, 14.73. Found: C,
C, 71.99; H, 4.03; N, 15.26. Found: C, 71.86; H, 4.16; N, 15.09%.
8-Benzoyl-7H-9-(4-methylphenylhydrazono)-pyrrolo[1,2-a]
perimidin-10-one (8a). Red solid, yield 1.04, 94.5%, mp
288–290ꢀC, (EtOH/Dioxane), IR ύ: 3444, 3205 (2NH), 1689,
1
1647 (2CO) cmÀ1, H NMR (DMSO-d6) 2.49 (s, 3H, CH3), 6.82
68.05; H, 3.50; N, 14.69%.
(d, J = 8 Hz, 2H, Ar H), 7.15 (d, J = 8 Hz, 2H, Ar H), 7.49–8.45
(m, 11H, Ar H) 11.89 (s, 1H, NH), 12.04 (s, 1H, OH); ms m/z
(%), 445 (M+, 7), 388 (3), 309 (4), 205 (4), 191 (3), 178 (4), 126
(12), 105 (22), 91 (7), 84 (16), 77 (22), 60 (100). Anal. Calcd for
C28H20N4O2 (444.46): C, 75.66; H, 4.53; N, 12.6. Found: C,
8-Benzoyl-7H-9-(4-nitrophenylhydrazono)-pyrrolo[1,2-a]
perimidin-10-one (8h). Red solid, yield 1.15, 97%, mp >310ꢀC,
(EtOH/Dioxane), IR ύ: 3380, 3200 (2NH), 1690, 1647 (2CO)
cmÀ1; ms m/z (%) 476 (M+ + 1, 30), 475 (M+, 43), 474 (32), 339
(34), 233 (23), 155 (30), 152 (34), 129 (32), 122 (16), 111 (43),
110 (36), 108 (39), 105 (100), 91 (39), 84 (31), 77 (93). Anal.
Calcd for C27H17N5O4 (475.44): C, 68.2; H, 3.6; N, 14.73.
Found: C, 68.03; H, 3.42; N, 14.53%.
75.48; H, 4.41; N, 12.49%.
8-Benzoyl-7H-9-(3-methylphenylhydrazono)-pyrrolo[1,2-a]
perimidin-10-one (8b). Orange solid, yield 1.01, 91%, mp
262–264ꢀC, (EtOH/Dioxane), IR ύ: 3420, 3220 (2NH), 1690,
Biological activity (sensitivity tests) by Kirby–Bauer
method. Antimicrobial activity of the tested samples was
determined using a modified Kirby–Bauer disc diffusion method
[29]. Briefly, 100 mL of the test bacteria/fungi were grown in
10mL of fresh media until they reached a count of approximately
108 cells/mL for bacteria or 105cells/mL for fungi [30]. Of
the microbial suspension, 100mL was spread onto agar plates
corresponding to the broth in which they were maintained.
Isolated colonies of each organism that might be playing a
pathogenic role should be selected from primary agar plates and
tested for susceptibility by disc diffusion method [31,32]. Of the
many media available, NCCL recommends Mueller–Hinton agar
because it results in batch-to-batch reproducibility. Disc diffusion
method for filamentous fungi was tested by using an approved
standard method (M38-A) developed by [33] for evaluating the
1647 (2CO) cmÀ1 1H NMR (DMSO-d6) 2.20 (s, 3H, CH3),
,
6.67–8.48 (m, 15H, Ar H), 11.85 (s, 1H, NH), 12.04 (s, 1H, OH);
ms m/z (%) 445 (M+ + 1, 23), 444 (M+, 68), 426 (23), 339 (31),
325 (5), 310 (28), 105 (80), 91(50), 77 (100). Anal. Calcd for
C28H20N4O2 (444.46): C, 75.66; H, 4.53; N, 12.6. Found: C,
75.50; H, 4.62; N, 12.38%.
8-Benzoyl-7H-9-phenylhydrazono-pyrrolo[1,2-a]perimidin-10-
one (8c). Orange solid, yield 1.02, 95%, mp 282–284ꢀC, (EtOH/
Dioxane), IR ύ: 3325, 3244 (2NH), 1693, 1647 (2CO) cmÀ1, 1H
NMR (DMSO-d6) 6.83–8.48 (m, 21H, ArH), 11.84 (s, 1H, NH),
12.11 (s, 1H, OH). ms m/z (%), 430 (M+, 50), 429 (33), 339
(13), 338 (13), 325 (20), 311 (16), 309 (14), 233 (17), 152
(10), 126 (7), 105 (56), 93 (21), 77 (100). Anal. Calcd for
C27H18N4O2 (430.46): C, 75.34; H, 4.21; N, 13.01. Found: C,
75.25; H, 4.05; N, 13.27%.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

T. A. Farghaly and H. K. Mahmoud
Vol 000
[8] Liu, K. C. Zhonghua Yaoxue Zazhi 1988, 40, 203.
susceptibility of filamentous fungi to antifungal agents. Disc
diffusion method for yeasts was developed by using an approved
standard method (M44-P) by [34]. Plates were inoculated with
filamentous fungi as A. flavus at 25ꢀC for 48h; Gram (+) bacteria
as S. aureus and Bacillus subtilis and Gram (À) bacteria as E. coli
and Pseudomonas aeuroginosa, which were incubated at 35–37ꢀC
for 24–48 h, and yeast as C. albicans incubated at 30ꢀC foe
24–48 h, and then the diameter of the inhibition zone was measured
in millimeters [29]. Standard disc of tetracycline (antibacterial
agent) and amphotericin B (antifungal agent) served as positive
control for antimicrobial activity, but filter discs impregnated with
10mL of solvent (distilled water, chloroform, DMSO) were used
as negative control. The agar used is Meuller–Hinton agar that is
rigorously tested for composition and pH. Further, the depth of the
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 discs (Schueicher and Schuel, Spain) with a diameter of
8.0 mm were impregnated with 10mL of the tested concentration of
the stock solutions. When a filter paper disc impregnated with a
tested chemical is placed on agar, the chemical will diffuse from
the disc into the agar. This diffusion will place the chemical in the
agar only around the disc. The solubility of the chemical and its
molecular size will determine the size of the area of chemical
infiltration around the disc. If an organism is placed on the agar, it
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chemical. This area of no growth around the disc is known as a
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Journal of Heterocyclic Chemistry
DOI 10.1002/jhet