Synthesis and antimicrobial activity of novel 5-amino-4-cyano-1H-pyrazole and quinazolin-4(3H)-one derivatives

Article abstract of DOI:10.1007/s12272-010-1202-5

New substituted aroylhydrazones (4a-f) were synthesized from the acid hydrazide (3) and the corresponding aldehyde or aldose. 5-Amino-4-cyano-1H- pyrazole derivatives (6a-f) were prepared by the reaction of the aroylhydrazones (4a-f) with malononitrile. T

Full text of DOI:10.1007/s12272-010-1202-5

Arch Pharm Res Vol 33, No 12, 1891-1900, 2010
DOI 10.1007/s12272-010-1202-5
Synthesis and Antimicrobial Activity of Novel 5-Amino-4-cyano-1H-
pyrazole and Quinazolin-4(3H)-one Derivatives
Mohamed T. Abdel-Aal1, Abdel-Aleem H. Abdel-Aleem1, Laila I. Ibahim2, and Ahmed L. Zein2
2
1Department of Chemistry, Faculty of Science, Menofeia University, Shebin-El-Koom 12345, Egypt and National Organi-
zation of Drug Control and Research, Cairo 11978, Egypt
(Received February 18, 2010/Revised May 17, 2010/Accepted May 23, 2010)
New substituted aroylhydrazones (4a-f) were synthesized from the acid hydrazide (3) and the
corresponding aldehyde or aldose. 5-Amino-4-cyano-1H-pyrazole derivatives (6a-f) were pre-
pared by the reaction of the aroylhydrazones (4a-f) with malononitrile. The synthesized com-
pounds were tested for antimicrobial activity against various bacteria and fungi and showed
moderate to high inhibition activities. Compounds incorporating a sugar moiety as well as a
pyrazolyl ring in their structure displayed the highest antimicrobial activity.
Key words: Aroylhydrazones, 5-Amino-4-cyanopyrazole, Quinazoline-4-one, Antimicrobial
activity
zide and hydrazones are claimed to exhibit appreci-
able antimicrobial activity (Eisa et al., 1991; Gursoy et
INTRODUCTION
Pyrazole containing structural moieties are of al., 1997; Rollas et al., 2002; Vicini et al., 2002). A num-
biological interest because they are the key structures ber of hydrazones have been used in the treatment of
in various therapeutic compounds (Anzai et al., 2004; several diseases such as tuberculosis, leprosy, and
Haddad et al., 2004). These compounds are known to mental disorders and they are also used as herbicides,
display antibacterial, antifungal (Tanitame et al., insecticides, rodenticides, and plant growth regulators
2004), anti-inflammatory, analgesic, and antipyretic (Alcock et al., 1972; Katyal and Dutta, 1975; Toth,
(Menozzi et al., 1992) activities. Pyrazole-containing 1981; El-Sayed et al., 2010).
compounds are also an essential structure in many
Quinazolin-4(3H)-ones are another important class
chemotherapeutic agents with potential antimicrobial of compounds that have widespread and diverse
(Foks et al., 2005; Dardari et al., 2006; Gilbert et al., pharmacological activities (Hori et al., 1990; Rewcastle
2006), antiparasitic (Rathelot et al., 2002; Bernardino et al., 1998). Quinazolin-4(3H)-one ring systems have
et al., 2006), antimalarial (Katiyar et al., 2005), and demonstrated antimalarial, antifungal, antitumor,
antiviral activities (Moukha-Chafiq et al., 2002; Allen anticonvulsant, and antihypertensive activities (Vaidya
et al., 2006). A previous investigation revealed that et al., 1983; Witt and Bergman, 2003; Connolly et al.,
some 5-amino-4-cyanopyrazole derivatives have anti- 2005). For example, albaconazole is one antifungal
bacterial activity (Spassova et al., 1980). For example, agent containing a quinazoline moiety in its structure.
sulfaphenazole, which incorporates a pyrazolyl ring These compounds are also useful in the treatment of
system, is a known potent antibacterial agent (Orth, Parkinson’s disease, depression, epilepsy, and diabetes
1968). Pyrazoles can be synthesized following various (Parmer and Singh, 1979).
synthetic routes and acid hydrazides are either useful
Nucleosides and their acyclic and
C-nucleoside ana-
intermediates or precursors for their synthesis. Hydra- logues possess a wide range of medicinal properties,
including antibacterial, antiviral, and antitumor acti-
vities (Larsson et al., 1983; Remy and Secrist, 1985;
Chu and Cutler, 1986; Holy, 1987; El Ashry and El
Kilany, 1996, 1997a, 1997b; Franchetti et al., 1997;
Hammerschmidt et al., 1997; Makara and Keseru,
Correspondence to: M. T. Abdel-Aal, Department of Chemistry,
Faculty of Science, Menoufia University, Egypt
Tel: 2-048-2235690, Fax: 2-048-2235689
E-mail: mtaha2000_2000@yahoo.com
1891

1892
M. T. Abdel-Aal et al.
1997). The therapeutic importance of these ring sys- reflux for 2 h. The solvent was removed under reduced
tems has triggered a renewed interest in the develop- pressure, and the solid residue that formed during
ment of new synthetic molecules formed by the combi- evaporation was filtered off, washed with water, dried,
nation of a pyrazole and a carbohydrate moiety. An and crystallized from ethanol to produce 2.92 g of
3
interest in the attachment of carbohydrate moieties to (78% yield) as white powder, m.p. 240-242oC; IR (KBr)
heterocycles and the synthesis of nucleoside analogues
has led us to develop new biologically active compounds (NH); H-NMR (DMSO-d6):
ν
cm-1: 1718 (C=O), 1718 (C=O), 1764 (C=O), 3254
1
δ 4.3 (bs, 2H, NH2), 6.92
(Abdel-Aal et al., 2003, 2006, 2008). Therefore, in the (m, 3H, Ar-3H), 7.32 (d, 2H, Ar-2H), 7.43 (m, 2H, Ar-
present work we aimed to synthesize new substituted 2H), 7.48 (d, 2H,
pyrazole and pyrimidine derivatives as well as to 2H), 7.92 (d, 2H,
evaluate their antimicrobial activity.
J
J
= 8.4 Hz, Ar-2H), 7.55 (m, 2H, Ar-
= 8.4 Hz, Ar-2H), 10.1 (s, 1H, NH),
10.6 (s, 1H, NH), 11.3 (s, 1H, NH); 13C-NMR (DMSO-
d6):
δ 167.44, 166.63, 164.63 (3CO), 141.05-120.05 (Ar-
carbons). Anal. calcd. for C21H18N4O3 (374.39): C,
67.37; H, 4.85; N, 14.96. Found: C, 67.14; H, 4.78; N,
MATERIALS AND METHODS
Melting points were determined using a Kofler block 14.80%.
instrument (BÜCHI Labortechnik AG). 1H-NMR spec-
tra were recorded with a Bruker AC 250 FT-NMR
spectrometer (FT-NMR; IBM Instruments Co.) at 250
MHz with TMS as an internal standard. The micro-
General procedure for the synthesis of aroyl-
hydrazone derivatives (4a-d)
A mixture of hydrazino compound
3 (1.87 g, 5 mmol)
analyses were performed at the Microanalytical Unit and the corresponding aldehyde (thiophene-2-carbox-
at Cairo University, Egypt, and were found to agree aldehyde, furan-2-carboxaldehyde, D-glucose, D-xylose,
favorably with the calculated values. The antimicro- D-galactose or D-arabinose) (5 mmol) in ethanol (20
bial activity was measured at the National Organiza- mL) with a catalytic amount of glacial acetic acid was
tion of Drug Control and Research, Cairo, Egypt. The heated under reflux for 2 h. After concentration and
starting compound 2-phenyl-1,3,4-benzoxazinone (
1)
cooling, the separated precipitate was filtered off,
was prepared from the reaction of anthranilic acid dried, and finally recrystallized from ethanol to afford
with benzoylchloride in the presence of dry pyridine the corresponding aroylhydrazone derivatives 4a-f
according to a previously reported procedure (Sammour (77-80% yields)
et al., 1971)
2-(Benzoylamino)-N-[4-(2-(thiophen-2-ylmethylene)
hydrazine)-carbonyl]phenylbenzamide (4a)
2-(Benzoylamino)-
(2)
N
-carbonyl-methylbenzoate
White powder (1.87 g, 80%), m.p. 250-252oC; IR (KBr)
A mixture of 2-phenyl-1,3,4-benzoxazinone (
g, 20 mmol) and of -aminomethylbenzoate (3.2 g, 20 6.82 (m, 4H, Ar-H), 6.94 (d, 2H,
mmol) in glacial acetic acid (15 mL) was heated under (m, 4H, Ar-H), 7.36 (d, 2H, = 8.4 Hz, Ar-H), 7.45 (m,
reflux for 6 h, cooled, and then the precipitate was 3H, Ar-H), 7.98 (d, 1H, = 8.4 Hz, Ar-H), 8.64 (s, 1H,
filtered and crystallized from glacial acetic acid to N=C ), 10.73 (s, NH), 11.42 (s, 1H, NH), 11.75 (s,
(75% yield) as a white powder, m.p. NH); 13C-NMR (DMSO-d6):
167.50, 164.66, 162.34
cm-1: 1660 (C=O), 1720 (C=O), (3C=O), 142.59 (C=N), 141.74-120.10 (Ar-carbons).
3.80 (s, 3H, CH3), Anal. calcd. for C26H20N4O3S (468.53): C, 66.65; H,
4.30; N, 11.96. Found: C, 66.52; H, 4.22; N, 11.85%.
1) (4.5
ν δ
cm-1: 1670 (C=O), 3250 (NH); 1H-NMR (DMSO-d6):
p
J = 7.4 Hz, Ar-H), 7.05
J
J
H
afford 5.61 g of
2
δ
220-221oC; IR (KBr)
ν
1
3220 (NH); H-NMR (DMSO-d6):
δ
6.55 (m, 2H, Ar-H), 6.72 (m, 3H, Ar-H), 6.91 (d, 2H,
= 8.4 Hz, Ar-H), 6.98 (m, 2H, Ar-H), 7.15 (m, 2H, Ar-
H), 7.48 (d, 2H, = 8.4 Hz, Ar-H), 10.82 (s, 1H, NH), 2-(Benzoylamino)-
11.35 (s, 1H, NH); 13C-NMR (DMSO-d6):
J
J
N-[4-[2-(furan-2-ylmethylene)
δ
167.54, hydrazino]-carbonyl]phenylbenzamide (4b)
165.72, 164.71 (3CO), 143.15-120.01 (Ar-carbons), 51.86
(CH3). Anal. calcd. for C22H18N2O4 (374.39): C, 70.58; H,
4.85; N, 7.48. Found: C, 70.27; H, 4.79; N, 7.39%.
White powder (1.74 g, 77%), m.p. 230-232oC; IR (KBr)
1
ν
cm-1: 1659 (C=O), 1716 (C=O), 3285 (NH); H-NMR
(DMSO-d6):
Hz, Ar-H), 7.12 (m, 1H, Ar-H), 7.54 (d, 2H,
Ar-H), 7.77 (d, 1H, = 7.8 Hz, Ar-H), 7.81 (m, 2H, Ar-
H), 7.81 (m, 2H, Ar-H), 7.88 (d, 2H, = 8.4 Hz, Ar-H),
(3.74 g 10 mmol) in etha- 8.05 (d, 2H, = 8.4 Hz, Ar-H), 8.34 (s, 1H, N=C ),
δ
6.91 (m, 3H, Ar-H), 6.93 (d, 1H,
J = 7.8
J
= 8.4 Hz,
J
2-(Benzoylamino)-N-carbonyl-benzoylhydrazide
J
(3)
To a solution of 1.87 g of
2
J
H
nol (20 mL), 2 g of hydrazine hydrate (0.5 g, 10 mmol) 10.55 (s, 1H, NH), 11.26 (s, 1H, NH), 11.62 (s, 1H,
was added. The reaction mixture was heated under NH). Anal. calcd. for C26H20N4O4 (452.46): C, 69.02; H,

Antimicrobial Activity of Pyrazole Derivatives
1893
Scheme 1
4.46; N, 12.38. Found: C, 70.14; H, 4.59; N, 12.21%.
1H, H-5), 3.75 (m, 2H, H-3,4), 3.86 (m, 1H, H-2), 4.27
(m, 2H, 2OH), 5.13 (d, 1H,
1H, = 5.2 Hz, OH), 5.13 (m, 1H, OH), 6.95 (m, 3H,
Ar-H), 7.49 (d, 2H,
J = 4.6 Hz, OH), 5.13 (t,
D-Glucose-2-(benzoylamino)-
4-carbonyl-hydrazone (4c)
N
-phenylbenzamide-
J
J
= 8.4 Hz, Ar-H), 7.53 (d, 2H, J =
Yellow powder (2.09 g, 78%), m.p. 210-211oC; IR 8.4 Hz, Ar-H), 7.85 (m, 2H, Ar-H), 7.88 (m, 2H, Ar-H),
1
(KBr)
ν
cm-1: 1960 (C=O), 3250 (NH), 3270 (OH); H- 7.89 (d, 1H,
J
= 8.2 Hz, H-1), 8.12 (d, 2H,
J
= 8.4 Hz,
NMR (DMSO-d6):
δ
3.41 (m, 2H, H-6, H-6), 3.63 (m, Ar-H), 10.52 (s, 1H, NH), 11.22 (s, 1H, NH), 11.65 (s,

1894
M. T. Abdel-Aal et al.
Scheme 2
1H, NH). Anal. calcd. for C27H28N4O8 (536.53): C, 60.44; NH), 11.75 (s, 1H, NH). Anal. calcd. for C26H26N4O7
H, 5.26; N, 10.44. Found: C, 60.31; H, 5.18; N, 10.38%. (506.18): C, 61.65; H, 5.17; N, 11.06. Found: C, 61.57;
H, 5.19; N, 10.96%.
D-Xylose-2-(benzoylamino)-N-phenylbenzamide-
4-carbonyl-hydrazone (4d)
D-Galactose-2-(benzoylamino)-N-phenylbenza-
White powder (1.95 g, 77%), m.p. 215-217oC; IR mide-4-carbonyl-hydrazone (4e)
1
(KBr)
NMR (DMSO-d6):
2H, H-3,4), 3.83 (m, 1H, H-2), 4.25 (m, 1H, OH), 5.13 NMR (DMSO-d6):
(d, 1H, = 5.6 Hz, OH), 5.13 (t, 1H, = 5.8 Hz, OH), 1H, H-5), 3.71 (m, 2H, H-3,4), 3.82 (m, 1H, H-2), 4.21
5.17 (m, 1H, OH), 6.55 (m, 3H, Ar-H), 6.88 (d, 1H, (m, 2H, 2OH), 5.13 (d, 1H, = 6.8 Hz, OH), 5.17 (t,
7.8 Hz, H-1), 7.56 (d, 2H, Ar-H), 7.62 (d, 2H, = 8.4 Hz, 1H, = 5.8 Hz, OH), 5.17 (m, OH), 6.78 (m, 3H, Ar-H),
Ar-H), 7.71 (m, 2H, Ar-H), 7.82 (m, 2H, Ar-H), 8.43 (d, 6.85 (m, 2H, Ar-H), 7.51 (d, 2H, = 8.4 Hz, Ar-H),
2H, = 8.4 Hz, Ar-H), 9.92 (s, 1H, NH), 10.63 (s, 1H, 7.85 (m, 2H, Ar-H), 7.87 (m, 2H, Ar-H), 7.89 (d, 1H,
ν
cm-1: 1690 (C=O), 3220 (NH), 3250 (OH); H-
White powder (2.12 g, 79%), m.p. 222-223oC; IR
1
δ
3.44 (m, 2H, H-5, H-5), 3.75 (m, (KBr)
ν
cm-1: 1690 (C=O), 3310 (NH), 3335 (NH); H-
δ
3.45 (m, 2H, H-6, H-6), 3.68 (m,
J
J
J
=
J
J
J
J
J
J

Antimicrobial Activity of Pyrazole Derivatives
1895
= 8.5 Hz, H-1), 8.15 (d, 2H,
J
= 8.4 Hz, Ar-H), 10.52 (s, 2H,
J
= 8.4 Hz, Ar-H), 7.65 (m, 3H, Ar-H), 7.72 (m,
1H, NH), 11.25 (s, 1H, NH), 11.66 (s, 1H, NH). Anal. 1H, Ar-H), 7.85 (m, 2H, Ar-H), 7.88 (m, 2H, Ar-H),
calcd. for C27H28N4O8 (536.53): C, 60.44; H, 5.26; N, 8.16 (d, 2H,
= 8.4 Hz, Ar-H), 10.67 (s, 1H, NH); 13C-
10.44. Found: C, 60.29; H, 5.16; N, 10.36%. NMR (DMSO-d6): 118.45-148.32 (Ar-carbons), 167.24,
J
δ
165.23, 164.53 (3C=O), 155.25 (CN). Analysis calcd. for
C29H20N6O4 (516.51): C, 67.44; H, 3.90; N, 16.27.
Found: C, 67.21; H, 3.79; N, 16.15%.
D-Arabinose-2-(benzoylamino)-
mide-4-carbonyl-hydrazone (4f)
N-phenylbenza-
White powder (1.97 g, 78%), m.p. 205-207oC; IR
1
(KBr)
NMR (DMSO-d6):
2H, H-3,4), 3.91 (m, 1H, H-2), 4.35 (m, 1H, OH), 5.25 (5c)
(d, 1H, = 6.8 Hz, OH), 5.39 (t, 1H, = 5.8 Hz, OH),
White powder (0.47 g, 78%), m.p. 219-220oC; IR
5.41 (m, 1H, OH), 6.95 (m, 3H, Ar-H), 7.14 (d, 1H, (KBr)
cm-1: 1690 (C=O), 2280 (CN), 3285 (NH), 3390
8.2 Hz, H-1), 7.51 (m, 2H, Ar-H), 8.66 (d, 2H, = 8.4 (OH); H-NMR (DMSO-d6):
Hz, Ar-H), 7.82 (m, 2H, Ar-H), 7.85 (m, 2H, Ar-H), 8.15 3.55 (m, 1H, H-4), 3.72 (m, 2H, H-2,3), 3.89-4.12 (m,
(d, 2H, = 8.4 Hz, Ar-H), 9.96 (s, 1H, NH), 10.63 (s, 1H, 2H, 2OH), 4.87 (s, 2H, NH2), 5.13 (d, 1H, = 5.4 OH),
NH), 11.75 (s, 1H, NH). Anal. calcd. for C26H26N4O7 5.13 (t, 1H, = 5.6 Hz, OH), 5.18 (m, 1H, H-4), 5.89 (d,
(506.18): C, 61.65; H, 5.17; N, 11.06. Found: C, 61.54; 1H, = 7.5 Hz, H-1), 6.92 (m, 3H, Ar-H), 7.15 (d, 2H,
= 8.4 Hz, Ar-H), 7.75 (m, 2H, Ar-H), 7.82 (m, 2H, Ar-
H), 7.85 (m, 2H, Ar-H), 8.16 (d, 2H, = 8.4 Hz, Ar-H),
ν
cm-1: 1690 (C=O), 3290 (NH), 3370 (NH); H- 2-(Benzoylamino)-
N
-[4-(5-amino-4-cyano-3-glu-
δ
3.39 (m, 2H, H-5, H-5), 3.88 (m, cosyl-1H-pyrazol-1-yl)carbonylphenyl]benzamide
J
J
J
=
ν
1
J
δ 3.48 (m, 2H, H-5, H-5),
J
J
J
J
H, 5.12; N, 10.95%.
J
J
10.88 (s, 1H, NH). Anal. calcd. for C30H28N6O8 (600.58):
C, 60.00; H, 4.70; N, 13.00. Found: C, 60.21; H, 4.79;
N, 12.89%.
General Procedure for the synthesis of substi-
tuted 5-amino-4-cyanopyrazole derivatives (5a-
d)
To a solution of aroylhydrazone 4a-d (1 mmol) and
malononitrile (0.07 g, 1 mmol) in absolute ethanol (10 2-(Benzoylamino)-
N
-[4-(5-amino-4-cyano-3-xylosyl-
-pyrazol-1-yl)carbonylphenyl]benzamide (5d)
White powder (0.41 g, 72%) m.p. 220-221oC; IR
cm-1: 1680 (C=O), 2250 (CN), 3370 (NH), 3420
mL) a catalytic amount of triethylamine was added
and then the mixture was heated under reflux for 6 h.
1H
The excess of solvent was removed by vacuum evapo- (KBr)
ν
1
ration, and the residue was chromatographed on a (OH); H-NMR (DMSO-d6):
silica gel using chloroform/methanol (90:10, v/v) as 3.87 (m, 2H, H-2,3), 4.47 (m, 1H, OH), 4.85 (s, 2H, NH2),
eluent to give 5-amino-4-cyanopyrazole derivatives 4.95 (m, 1H, H-1), 5.18 (d, 1H, = 5.6 Hz, OH), 5.54 (t,
5a-d in 72-78% yields. 1H, = 6.4 Hz, OH), 5.76 (m, 1H, OH), 6.93 (m, 3H,
Ar-H), 7.76 (d, 2H, = 8.4 Hz, Ar-H), 7.80 (m, 2H, Ar-
-[4-(5-amino-4-cyano-3-thio- H), 7.91 (m, 2H, Ar-H), 8.56 (d, 2H, = 8.4 Hz, Ar-H),
δ 3.39 (m, 2H, H-4, H-4),
J
J
J
2-(Benzoylamino)-
N
J
phen-2-yl-1H-pyrazol-1-yl)carbonylphenyl]benza- 10.39 (s, 1H, NH). Anal. calcd. for C29H26N6O7 (570.55):
mide (5a)
C, 61.05; H, 4.59; N, 14.73. Found: C, 61.27; H, 4.71;
White powder (0.4 g, 75%), m.p. 265-266oC; IR (KBr) N, 14.59%.
ν
cm-1: 1664 (C=O), 2220 (CN), 3430 (NH), 3457 (NH2);
1H-NMR (DMSO-d6):
Ar-H), 6.95 (d, 1H,
8.4 Hz, Ar-H), 7.44 (m, 3H, Ar-H), 7.49 (d, 1H,
Hz, Ar-H), 7.86 (m, 2H, Ar-H), 7.88 (m, 2H, Ar-H), g, 10 mmol) and
8.05 (d, 2H,
= 8.4 Hz, Ar-H), 11.78 (s, 1H, NH). Anal. mmol) was fused at 160oC for 1.5 h, then the tempera-
calcd. for C29H20N6O3S (532.57): C, 65.40; H, 3.79; N, ture was elevated to 220oC until the evolution of water
δ
5.12 (s, 2H, NH2), 6.72 (m, 3H,
= 7.8 Hz, Ar-H), 7.38 (d, 2H,
= 7.8
4-[4-Oxoquinazolin-2-phenyl-3(4H)-yl]-N-methyl-
J
J =
benzoate (6)
A mixture of 2-phenyl-1,3,4-benzoxazinone (
J
1) (2.23
p-aminomethylbenzoate (1.51 g, 10
J
15.78. Found: C, 65.22; H, 3.65; N, 15.59%.
vapor ceased. The reaction mixture was left to cool, a
few drops of methanol were added, and it was left to
stand overnight. The separated solid was successively
purified using column chromatography (methylene
2-(Benzoylamino)-N-[4-(5-amino-4-cyano-3-furan-
2-yl-1
(5b)
H-pyrazol-1-yl)carbonylphenyl]benzamide
chloride/methanol) to afford 2.67 g of
6 (75%) as a
White powder (0.38 g, 74%), m.p. 245-246oC; IR white powder, m.p. 170-171oC; IR (KBr)
ν
cm-1: 1675
cm-1: 1660 (C=O), 2260 (CN), 3280 (NH), 3390 (C=O), 1748 (C=O); 1H-NMR (DMSO-d6):
ν δ 3.95 (s, 3H,
1
(KBr)
(NH2); H-NMR (DMSO-d6):
(m, 3H, Ar-H), 6.97 (d, 1H,
δ
4.85 (s, 2H, NH2), 6.9 CH3), 6.66 (m, 2H, Ar-H), 6.78 (m, 3H, Ar-H), 7.80 (m,
= 8.2 Hz, Ar-H), 7.58 (d, 2H, Ar-H), 6.92 (d, 2H, = 8.4 Hz, Ar-H), 7.18 (m, 2H,
J
J

1896
M. T. Abdel-Aal et al.
Ar-H), 7.45 (d, 2H,
C22H16N2O3 (356.37): C, 74.15; H, 4.53; N, 7.86. Found:
C, 74.11; H, 4.48; N, 7.69%.
J = 8.4 Hz, Ar-H). Anal. calcd. for
Antimicrobial activity
Growth media
Nutrient agar medium used in the evaluation of
antibacterial activity consisted of (g/L): peptone 5.0 g,
beef extract 3.0 g, sodium chloride 5.0 g, glucose 10.0
g, agar 18.0 g and water 1000 mL.
4-[4-Oxoquinazolin-2-phenyl-3(4
ylhydrazide (7)
H)-yl]-N-benzo-
Hydrazine hydrate (0.5 g, 10 mmol) was added to a
solution of compound
Sabouraud Dextrose agar medium used in the
(3.56 g, 10 mmol) in ethanol evaluation of antifungal activity consisted of (g/L):
6
(20 mL). The reaction mixture was heated under mycological peptone 10.0 g, glucose 40.0 g. Agar 15.09
reflux for 2 h. The solvent was evaporated under a and water 1000 mL with pH 5.0. Yeast extract medium
vacuum; the resulting solid was filtered off, washed used to measure anti-yeast activity consisted of (g/L):
with water, and then recrystallized from ethanol to yeast extract 400 g, malt extract 10.0 g, glucose 4.0 g
afford 2.67 g (75%) of hydrazide
m.p. 210-211oC; IR (KBr) cm-1: 1560 (C=O), 1720 claved in 121oC for 30 min under 1.56 atp.
(C=O), 3420 (NH); 1H-NMR (DMSO-d6):
3.95 (s, 3H, The antimicrobial activities of the compounds were
CH3), 4.88 (s, 2H, NH2), 6.66 (m, 2H, Ar-H), 6.81 (m, tested using the disc plate technique as described in
3H, Ar-H), 6.95 (d, 2H, = 8.4 Hz, Ar-H), 7.15 (m, 2H, British Pharmacopeia (1968). The specific media used
Ar-H), 7.19 (m, 2H, Ar-H), 7.56 (d, 2H,
= 8.4 Hz, Ar- for each test organism was melted, cooled to 45oC, and
7 as a white powder, and water 1000 mL with pH 7.3. All media were auto-
ν
δ
J
J
H), 12.55 (s, 1H, NH). Anal. calcd. for C21H16N4O2: C, then incubated with 1 mL of the bacteria, fungi. The
70.77; H, 4.53; N, 15.72. Found: C, 70.57; H, 4.42; N, flask was shaken well and the suspension was poured
15.40%.
into a 10 cm Petri dish. Five 6 mm disks were dipped
in different concentrations of compound solution (10,
100, 1000 µg), the solvent was allowed to evaporate,
and the disks were placed upon the surface incubated
with medium, which were kept in a refrigerator for 1
4-[4-Oxoquinazolin-2-phenyl-3(4H)-yl]-N-[thio-
phen-2-ylmethylene]-benzohydrazone (8)
Method A
To a solution of hydrazide
7 (1.78 g, 5 mmol) and h to permit diffusion of the antimicrobial substance.
thiophene-2-carboxaldehyde (0.56 g, 5 mmol) in etha- The plates were incubated at 37oC for 24 h for the
nol (20 mL) glacial acetic acid (1 mL) was added. The bacteria and 28oC for the fungi and yeast. The clear
mixture was heated under reflux for 2 h and then zone of inhibition was measured in mm, for each test
concentrated under reduced pressure. The separated the main values of inhibition zones were read in tri-
solid product was filtered off and crystallized from plicate and the calculated results were classified as
ethanol to give 1.57 g of
8
(70% yield)
follows:
No clearing zone: inactive
Method B
Small clearing zone: slightly active 12 mm
Medium clearing zone: moderately active 18 mm
Large clearing zone: highly active 30 mm
Aroylhydrazone 4a (2.34 g, 5 mmol) was added
portion wise to phosphorous oxychloride (10 mL) at
0oC with stirring for 1 h. The temperature was ele-
vated gradually to 60oC and kept at this tempera-
ture for 2 h. After cooling, the reaction mixture was
poured onto crushed ice and the solid product was
filtered off, washed with 5% NH4OH and water, and
RESULTS AND DISCUSSION
The reaction of the benzoxazinone derivative
-aminomethylbenzoate in acetic acid at reflux tem-
perature did not give the expected oxaquinazoline de-
(68% yield) as a white powder, m.p. 223-225oC; IR rivative; instead 2-benzoylamino-
-carbonylmethyl-
(KBr) ) was formed. The IR spectrum showed two
cm-1: 1684 (C=O), 3409 (NH); 1H-NMR (DMSO- benzoate (
d6): 3.95 (s, 3H, CH3), 4.88 (s, 2H, NH2), 6.66 (m, 2H, characteristic absorption bands in the carbonyl fre-
Ar-H), 6.81 (m, 3H, Ar-H), 6.95 (d, 2H,
= 8.4 Hz, Ar- quency region at 1660 and 1720 cm-1, which correspond
H), 6.97 (d, 1H, = 8.2 Hz, Ar-H), 7.31 (d, 2H, = 8.4 to the carbonyl amide and ester groups, respectively.
Hz, Ar-H), 7.45 (d, 1H, Ar-H), 7.49 (m, 3H, Ar-H), 7.56 The H-NMR spectrum showed peaks corresponding
1 with
p
then crystallized from ethanol to afford 1.53 g of
8
N
ν
2
δ
J
J
J
1
(d, 2H,
J = 8.4 Hz, Ar-H), 8.25 (s, 1H, N=CH), 12.5 (s, to a methyl group, the NH groups, and the aromatic
1H, NH). Anal. calcd. for C26H18N4O2S (450.51): C, protons. The 13C-NMR spectrum showed a methyl
69.32; H, 4.03; N, 12.44. Found: C, 69.17; H, 3.91; N, ester peak in addition to the aromatic carbons. Treat-
12.32%.
ment of 2-benzoylamino-N-carbonylmethylbenzoate
(2) with hydrazine hydrate in ethanol at reflux tem-

Antimicrobial Activity of Pyrazole Derivatives
1897
perature gave 2-benzoylamino-
drazide (
vative (
carboxaldehyde, D-glucose, D-arabinose, D-xylose, and mental conditions. Interestingly, the reaction of com-
D-galactose in ethanol and a catalytic amount of gla- pound with -aminomethylbenzoate in an oil bath at
cial acetic acid afforded the corresponding hydrazones 160ºC afforded 4-(4-oxoquinazolin-2-phenyl-3(4H)-yl)-
N-carbonylbenzoylhy-
In order to obtain the desired quinazoline derivative
3
). The reaction of the benzoylhydrazide deri- from the reaction of the benzoxazinone derivative (
1)
3
) with thiophene-2-carboxaldehyde, furan-2- with -aminomethylbenzoate we modified the experi-
p
1
p
1
(
4a-f). The structures of the hydrazones (4a-f) were
well characterized by the spectral data of IR, 1H-NMR, compound showed the protons of the methyl ester at
13C-NMR, and elemental analysis. The assignment of 4.14 ppm and aromatic protons in the range of
6.15-
NH and OH groups in these compounds was deter- 7.15 ppm. Hydrazinolysis of the quinazoline methyl
mined by D2O exchange. The IR spectra of the sugar ester derivative ( ) in refluxing ethanol gave the hy-
hydrazones (4c-f) showed that the hydroxyl groups of drazide compound
the sugar moiety were in the range of 3250-3420 cm-1. acid hydrazide showed the presence of a peak corres-
N-methylbenzoate (6). The H-NMR spectrum of this
δ
δ
7
1
7
. The H-NMR spectrum of this
The 1H-NMR spectra of these compounds showed that ponding to the NH2 at
the sugar protons were in the range of 3.45-5.65 and protons in the range of
that H-1 was in the range of 7.65-8.11 ppm. In the the hydrazide (
reaction of the 5-amino-4-aroylhydrazone derivatives ethanol in the presence of few drops of glacial acetic
4a-f) with malononitrile in ethanol the corresponding acid produced the hydrazone compound . In addition,
5-amino-4-cyano-3-arylpyrazole derivatives (5a-d) were compound was also successfully obtained by stirring
obtained after column chromatography purification. compound 4a with phosphorous oxychloride at 60oC.
The IR spectra of these compounds showed a charac- The structure of the hydrazone derivative synthe-
δ
4.7 ppm as well as aromatic
δ 7.1-8.6 ppm. The reaction of
δ
δ
7) with thiophene-2-carboxaldehyde in
(
8
8
8
teristic absorption band corresponding to the CN sized by these two different methods was confirmed by
1
group as well as the carbonyl group band. The 1H and IR, H-NMR spectra, and elemental analysis, which
13C spectra of these compounds were in full agreement were in agreement with the assigned structure.
with their assigned structures.
Table I. Antibacterial activity of newly synthesized compounds
Gram negative bacteria
Salmonella Escherichia coli
Gram positive bacteria
Staphylococcus aureus Bacillus subtilis
Compound
Cc
Cb
Ca
Cc
Cb
Ca
Cc
Cb
Ca
Cc
Cb
Ca
2
3
+
++
++
++
+
++
++
+
+
+
+
+
+
+
+
-
-
+
+
+
+
+
+
+
+
-
+
+
-
+
+
+
+
+
+
-
-
+
-
+
+
-
+
++
++
++
+++
++
++
+
+
+
+
-
++
+
+
+
-
+
+
++
+
+
+
+
+
+++
-
+
-
+
+++
+
+
+
+
+
-
-
-
-
-
-
-
-
+
+
-
+
+
+
+
-
+
-
-
+
-
+
+
+
+++
-
++
+
++
+++
++
++
+
+++
++
++
+
++
+
-
+
-
+
+++
+
+
+
+
+
-
-
-
-
-
-
-
-
++
++
++
++
++
++
+
+++
++
++
++
++
+
4a
4b
4c
4d
4e
4f
5a
5b
5c
5d
5e
5f
6
+
++
+
+
+
+
+
-
-
+
-
+
-
+
++
+
++
+
+
+
+
+
-
-
+
-
+
-
+
++
+
+
+
++
++
+
++
+
++
-
++
++
++
+++
++
+
++
+
++
+
++
+++
+
-
+
+++
++
+
++
+++
+
-
+
+++
7
8
++
+++
Sulfaphenazole +++
+++
++
Ca Activity of synthesized compounds in concentration of 10 ppm.
Cb Activity of synthesized compounds in concentration of 100 ppm.
Cc Activity of synthesized compounds in concentration of 1000 ppm.
- No antimicrobial effect
+ Low antimicrobial effect (4 mm)
++ Moderate antimicrobial effect (8-10 mm)
+++ High antimicrobial effect (15-18 mm)

1898
M. T. Abdel-Aal et al.
wide spectrum of activity, especially at concentrations
Antimicrobial activity
The newly synthesized compounds were tested for of 100 and 1000 ppm.
their antimicrobial action against Gram positive bac-
These results and the structure activity relationship
teria (Bacillus subtilis and Staphylococcus aureus), suggest that the introduction of a sugar moiety as well
Gram negative bacteria (Escherichia coli and Salmo- as a pyrazolyl ring into the quinazolin-4(3H)-one
nella), two fungal strains (Aspergillus flavus and As- structure enhances the antimicrobial activity against
pergillus niger), and one yeast strain (Candida albi- both Gram positive and Gram negative bacteria even
cans). Most of the synthesized products showed high at the lower concentrations. Both the acyclic nucleo-
bactericidal, fungicidal, and anti-yeast activity. The side analogous 5c and 5d of the substituted pyrazole
antibacterial and antifungal activities were studied nucleobase displayed high inhibition activity against
using sulfaphenazole and albaconazole as positive Gram negative bacteria (Escherichia coli). The most
controls, respectively.
effective derivatives have the carbon ring substituted
The results of antimicrobial screening revealed that with a methyl or methoxy group. Consequently, it is
compound 4c showed the highest activity against both expected that they could be the basis for further re-
types of bacteria (Table I and II). Compounds
4c 4d, and 4f were found to be active against both
Gram positive and Gram negative bacteria, whereas amino-4-cyano-1
compound 5a was only active against Gram positive zoline-4-one derivatives were synthesized and evalu-
bacteria and compound was only active against ated for their antimicrobial activity. Compounds in-
3,
4a
,
search.
In conclusion, new substituted aroylhydrazones, 5-
-pyrazole, and -substituted quina-
,
H
N
2
Gram negative bacteria. Most of the synthesized com- corporating a sugar moiety as well as a pyrazolyl ring
pounds did not show activity against fungi or yeast in their structure displayed high antimicrobial activity
except for compounds 5c
,
5d and
8
that showed a against both Gram positive and Gram negative bac-
teria.
Table II. Antifungal activity of newly synthesized com-
pounds
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-
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