5-Nitroimidazole derivatives as possible antibacterial and antifungal agents

Article abstract of DOI:10.1016/S0014-827X(99)00109-3

Some novel 1-[2-[[5-(2-furanyl)-4-substituted 4H-1,2,4-triazol-3- yl]thio]ethyl]-2-methyl-5-nitro-1H-imidazoles (3), 1-[3-[[5-(2-furanyl/2- thienyl)-4-substituted 4H-1,2,4-triazol-3-yl]-thio]-2-hydroxypropyl]-2- methyl-5-nitro-1H-imidazoles (5) and 1-[3-[(N,N-disubstituted thiocarbamoyl)- thio]-2-hydroxypropyl]-2-methyl-5-nitro-1H-imidazoles (7) were synthesized and evaluated for in vitro antibacterial and antifungal activity. Some of 5 were found to be effective against bacteria and fungi (minimum inhibitory concentration (MIC) 7.3-125 μg/ml), whereas 7 were found to be effective against fungi (MIC 3-25 μg/ml).

Full text of DOI:10.1016/S0014-827X(99)00109-3

www.elsevier.nl/locate/farmac
Il Farmaco 54 (1999) 826–831
5-Nitroimidazole derivatives as possible antibacterial and
antifungal agents
a
Nur Sibel Gu¨nay a, Gu¨ltaze C¸ apan a,*, Nuray Ulusoy a, Nedime Ergenc¸ ,
b
c
8
Gu¨lten Otu¨k , Dilek Kaya
a Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Uni6ersity of Istanbul, 34452 Istanbul, Turkey
b Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Uni6ersity of Istanbul, 34452 Istanbul, Turkey
c Department of Microbiology, Istanbul Faculty of Medicine, Uni6ersity of Istanbul, 34390 Istanbul, Turkey
Received 24 March 1999; accepted 15 September 1999
Abstract
Some novel 1-[2-[[5-(2-furanyl)-4-substituted 4H-1,2,4-triazol-3-yl]thio]ethyl]-2-methyl-5-nitro-1H-imidazoles (3), 1-[3-[[5-(2-
furanyl/2-thienyl)-4-substituted 4H-1,2,4-triazol-3-yl]-thio]-2-hydroxypropyl]-2-methyl-5-nitro-1H-imidazoles (5) and 1-[3-[(N,N-
disubstituted thiocarbamoyl)-thio]-2-hydroxypropyl]-2-methyl-5-nitro-1H-imidazoles (7) were synthesized and evaluated for in
vitro antibacterial and antifungal activity. Some of 5 were found to be effective against bacteria and fungi (minimum inhibitory
concentration (MIC) 7.3–125 mg/ml), whereas 7 were found to be effective against fungi (MIC 3–25 mg/ml). © 1999 Elsevier
Science S.A. All rights reserved.
Keywords: 5-Nitroimidazoles; 1,2,4-Triazoles; Dithiocarbamates; Antibacterial activity; Antifungal activity
sis and antimicrobial evaluation of 1,2,4-triazoles and
dithiocarbamic acid esters [12–14], some new nitroimi-
1. Introduction
dazole derivatives bearing a 1,2,4-triazolylthioethyl,
1,2,4-triazolylthiopropyl or N,N-disubstituted dithio-
carbamoyl–propyl residue at N-1 were synthesized and
Metronidazole and related N-1 substituted 5-nitroim-
idazoles like ornidazole, secnidazole and tinidazole are
widely used in the treatment of diseases caused by
evaluated for in vitro antibacterial and antifungal
protozoa and anaerobic bacteria [1–3]. Furthermore,
activity.
some 5-nitroimidazoles have been shown to sensitize
hypoxic tumor cells to the effects of ionizing radiation
[4]. It has been speculated that a reactive intermediate
formed in the microbial reduction of the 5-nitro group
of nitroimidazoles covalently binds to the DNA of the
2. Chemistry
microorganism, triggering the lethal effect. Potential
reactive intermediates include the nitroxide, nitroso,
hydroxylamine and amine. The ability of nitroimida-
1-(2-Chloroethyl)-2-methyl-5-nitro-1H-imidazole (1)
and 1-(3-chloro-2-hydroxypropyl)-2-methyl-5-nitro-1H-
imidazole (ornidazole, 4) was reacted with the anion
zoles, especially metronidazole, to act as radiosensitiz-
ing agents is also related to their reductive potentials.
1,2,4-Triazoles and dithiocarbamic acid esters are also
reported to show antifungal and antibacterial activity
[5–9]. In view of the wide continued interest in the
activity spectrum and profile of the nitroimidazoles
[10,11] and in continuation of our work on the synthe-
generated from 5-(2-furanyl/2-thienyl)-4-substituted
2,4-dihydro-3H-1,2,4-triazole-3-thiones (2) [12,15,16] in
the presence of K2CO3, to afford 1-[2-[[5-(2-furanyl)-4-
substituted 4H-1,2,4-triazol-3-yl]thio]ethyl]-2-methyl-5-
nitro-1H-imidazoles (3) and 1-[3-[[5-(2-furanyl/2-
thienyl)4-substituted 4H - 1,2,4 - triazol - 3 - yl]thio] - 2-
hydroxypropyl]-2-methyl-5-nitro-1H-imidazoles (5), re-
spectively. Treatment of 4 with potassium N,N-disub-
stituted dithiocarbamates 6 [14] furnished 1-[3-[(N,N-
disubstituted thiocarbamoyl)thio]-2-hydroxypropyl]-2-
* Corresponding author. Fax: +90-212-519-0812.
E-mail address: gulcapan@superonline.com (G. C¸ apan)
0014-827X/99/$ - see front matter © 1999 Elsevier Science S.A. All rights reserved.
PII: S0014-827X(99)00109-3

N.S. Gu¨nay et al. / Il Farmaco 54 (1999) 826–831
827
Table 1
methyl-5-nitro-1H-imidazoles (7) (Scheme 1, Table 1).
Physical constants of compounds 3, 5 and 7
Analytical (CHN) and spectral data (IR, 1H NMR,
EIMS) supported the designed structures.
The OꢀH (3446–3188 cm−1), ꢁCꢀH (3176–3096
cm−1), CꢁC/CꢁN (1610–1421 cm−1) and NO2 (1570–
1515 cm−1, 1366–1358cm−1) stretching vibrations ob-
served in the IR spectra provided substantial proof for
the formation of the desired products 3, 5 and 7.
The 1H NMR spectra of 3 displayed the NꢀCH2
resonances at about l 4.67–4.70 ppm as triplets. The
SꢀCH2 group in 3a was observed as a multiplet to-
gether with the NꢀCH3 singlet at about l 3.56–3.68
ppm while the SꢀCH2 protons of 3d and 3e resonated
as triplets at about l 3.60–3.62 ppm. Compounds 5
and 7 are chiral molecules, thus the geminal hydrogens
on the methylene groups adjacent to the chiral center
experience different magnetic environments. In rigid
systems where averaging is not possible, the geminal
1
hydrogens absorb at different l values on the H NMR
scale and show large couplings in the range of 15 Hz,
which are characteristic of geminal interactions [17].
Thus, the NꢀCH2 protons at N-1 of the imidazole
moiety, both in 5 and 7 absorbed as double doublets
Scheme 1. Synthetic routes to compounds 3, 5 and 7.

828
N.S. Gu¨nay et al. / Il Farmaco 54 (1999) 826–831
3. Results and discussion
centered at about l 4.23–4.56 ppm with coupling con-
stants in the ranges 14.1–13.7 and 9.4–2.5 Hz. The
SꢀCH2 protons also adjacent to the chiral center ab-
sorbed as double doublets centered at about l 3.38–
3.39 ppm due to magnetic unequivalence, but showed
smaller coupling constants in 5 (5.8–5.6 and 2.1–1.7
Hz) where the rotation about the SꢀCH2 bond was not
as much impeded as it was about the NꢀCH2 bond,
which is in direct relation with the rigid imidazole
system. The large coupling observed for the SꢀCH2
protons in 7 (l 3.59 ppm; 13.6 and 4.8 Hz) implicated
restriction of rotation about the SꢀC bond of the
dithiocarbamate moiety. The variation in the vicinal
coupling constants observed for these protons was at-
tributed to the variation in the dihedral angle between
the NꢀCH2 and SꢀCH2 protons and the CHOH proton.
The absorption positions and splitting patterns of the
other protons were in accordance with the literature
[12,17,18].
The molecular ions at m/z 444, 421 and 334 observed
in the EIMS of representative examples, (3a, 5i and 7b),
provided further evidence for the formation of
the expected structures. The fragmentation routes
primarily involved losses of OH (m/z 17), NO (m/z 30),
NO2 (m/z 46) and HNO2 (m/z 47) from the molecular
ion, which are characteristic of compounds carrying
OH and NO2 functions. Diagnostic fragments derived
from the imidazole and 1,2,4-triazole or N,N-
disubstituted dithiocarbamate systems were in accor-
dance with the structures reported in the literature
[14,19–21].
Compounds 3, 5 (except 5j and 5k) and 7 were
evaluated for antibacterial and/or antifungal activity
against representative bacteria — Staphylococcus au-
reus ATCC 6538, S. epidermidis ATCC 12228, Es-
cherichia coli ATCC 8739, Klebsiella pneumoniae UC
57, Shigella flexneri, Pseudomonas aeruginosa ATCC
1539, Proteus mirabilis and Salmonella typhi; and fungi
— Trichophyton tonsurans NCPF 245, T. mentagro-
phytes var. erinacei ATCC 375, T. mentagrophytes,
Microsporum gypseum NCPF 580, M. canis and M.
audouinii [22,23]. As can be seen in Table 2, although
not as active as the standard ampicillin, 5a–i were
found to be active against S. aureus ATCC 6538 and/
or S. epidermidis ATCC 12228 where ornidazole was
devoid of activity. These results indicate that incor-
poration of the 3-[[5-(2-furanyl)-4-substituted 4H-1,2,4-
triazol-3-yl]thio]-2-hydroxypropyl moiety imparts activ-
ity against aerobic bacteria since 3, which differs
from 5a–i in bearing an ethyl instead of the 2-hydrox-
ypropyl residue between the imidazole and 1,2,4-tria-
zole systems; 5l and 5m, which bear the 2-thienyl
instead of the 2-furanyl moiety at the 5-position of the
1,2,4- triazole ring; and 7, which does not bear the
1,2,4-triazole nucleus were found to be devoid of activ-
ity. Table 2 also contains results of antifungal activity
tests of 5 and 7 where miconazole was used as the
standard. The most active compound was 7a (T. ton-
surans NCPF 245; minimum inhibitory concentration
(MIC) 3 mg/ml) exerting about one half the activity
of onidazole. Although the rest of the compounds
Table 2
Antimicrobial activity of 5 and 7 (MIC mg/ml)
Microorganism a(MICmg/ml)
Comp.
A
B
C
D
E
F
G
H
5a
5b
5c
5d
5e
5f
5g
5h
58.6
58.6
58.6
29.2
7.3
29.2
14.6
29.2
14.6
n.a.
n.a.
n.a.
n.t.
n.a.
n.a.
n.a.
62.5
125
31.2
62.5
3.66
62.5
n.a.
n.a.
n.a.
n.t.
12.5
25
25
25
25
25
25
25
25
3
12.5
25
25
\25
25
25
\25
25
25
25
\25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
\25
\25
\25
25
\25
\25
\25
\25
\25
25
25
25
25
25
25
25
25
25
25
25
25
5I
7a
7b
25
1.6
0.2
n.t.
25
0.8
0.2
n.t.
Ornidazole
Miconazole
Ampicillin
6.2
0.2
n.t.
6.2
0.2
n.t.
6.2
0.2
n.t.
6.2
0.2
n.t.
0.06
1.95
a A=S. aureus ATCC 6538, B=S. epidermidis ATCC 12228, C=T. tonsurans NCPF 245, D=T. mentagrophytes var. erinacei ATCC 375,
E=T. menta-grophytes, F=M. gypseum NCPF 580, G=M. canis, H=M. audounii. n.a.=not active, n.t.=not tested.

N.S. Gu¨nay et al. / Il Farmaco 54 (1999) 826–831
829
showed varying degrees of inhibition (MIC 12.5–25
mg/ml), none were as effective as miconazole (MIC
0.2 mg/ml) or ornidazole (MIC 0.8–6.2 mg/ml).
3H, ar.), 7.81 (s, 1H, furan C5ꢀH), 8.02 (s, 1H, imida-
zole C4ꢀH).
Spectral data for 3e. IR [w cm−1, KBr]: 3105 (ꢁCꢀH),
1527 (NO2), 1513, 1493, 1464, 1421 (CꢁN/CꢁC), 1364
(NO2). 1H NMR [200 MHz, l ppm, DMSO-d6]: 2.41 (s,
3H, CH3), 3.60 (t, J=6.4 Hz, 2H, SꢀCH2), 4.69 (t,
J=6.4 Hz, 2H, NꢀCH2), 6.16 (d, J=3.4 Hz, 1H, furan
C3ꢀH), 6.51 (dd, J=3.4, 1.6 Hz, 1H, furan C4ꢀH), 7.29
(d, J=8.3 Hz, 2H, ar.), 7.41 (d, J=8.3 Hz, 2H, ar.),
7.75 (s, 1H, furan C5ꢀH), 7.96 (s, 1H, imidazole C4ꢀH).
4. Experimental
4.1. Chemistry
Melting points were determined with a Bu¨chi (Tot-
toli) melting point apparatus in open capillaries and are
1
uncorrected. IR, H NMR and EIMS were recorded on
4.1.3. Synthesis of 1-[3-[[5-(2-furanyl/2-thienyl)-4-
substituted 4H-1,2,4-triazol-3-yl]thio]-2-hydroxy-
propyl]-2-methyl-5-nitro-1H-imidazoles (5a–m)
To a solution/suspension of 2 (0.005 mol) in
CH3COCH3 (30 ml), 4 (0.005 mol) and K2CO3 (0.02
mol) were added. The reaction mixture was refluxed for
21 h, cooled and poured into H2O. The precipitate was
collected by filtration.
Shimadzu 2100S, Perkin–Elmer 1600 FTIR, Bruker
AC 200 (200 MHz) and VG Zab Spec (EI, 70 eV)
instruments, respectively. The reactions were monitored
by TLC (Silica Gel 60 F254 Merck Art. 5735).
4.1.1. Synthesis of 1-(2-chloroethyl)-2-methyl-5-nitro-
1H-imidazole (1)
To a suspension of 1-(2-hydroxyethyl)-2-methyl-5-ni-
tro-1H-imidazole (0.03 mol) in benzene (20 ml), SOCl2
(4.5 ml) was added and the reaction mixture was heated
under reflux for 2 h. After cooling the solvent was
decanted and the remaining crystalline precipitate was
washed with petroleum ether, neutralized by the addi-
tion of saturated NaHCO3 solution, washed with H2O
and filtered to afford 1, which was used without further
purification.
Compounds 5a, 5c, 5f, 5h, 5l and 5m were purified by
washing with H2O; 5b, 5e, 5i, 5j and 5k by recrystalliza-
tion from C2H5OH; 5d from C2H5OH:H2O and 5g by
washing with ether.
Spectral data for 5i. IR [w, cm−1, KBr]: 3188 (OH),
3096 (ꢁCH), 1603 (CꢁN/CꢁC), 1515 (NO2), 1463, 1430
1
(CꢁN/CꢁC), 1362 (NO2). H NMR [200 MHz, l ppm,
DMSO-d6]: 2.34 (s, 3H, CH3), 3.39 (dd, J=5.8, 2.1 Hz,
2H, SꢀCH2), 4.04–4.12 (m, 1H, CH), 4.23 (dd, J=
13.7, 9.2 Hz, 1H, NꢀCH2), 4.55 (dd, J=13.7, 2.5 Hz,
1H, NꢀCH2); 5.58 (d, J=5.3 Hz, 1H, OH); 6.27 (d,
J=3.3 Hz, 1H, furan C3ꢀH), 6.53 (dd, J=3.4, 2.0 Hz,
1H, furan C4ꢀH), 7.47 (d, J=8.5 Hz, 2H, ar.), 7.56 (dd,
J=9.0, 4.8 Hz, 2H, ar.), 7.74 (d, J=1.1 Hz, 1H furan,
C5ꢀH), 7.99 (s, 1H, imidazole C4ꢀH). EIMS [m/z (%)]:
444 (M+, 53), 427 (77), 414 (52), 398 (40), 397 (83), 384
(45), 366 (74), 354 (58), 334 (60), 316 (78), 305 (45), 288
(49), 286 (75), 274 (88), 244 (66), 230 (84), 229 (55), 202
(85), 169 (90), 147 (70), 134 (82), 132 (80) 120 (84), 104
(88), 98 (100), 93 (20), 78 (86), 65 (90).
4.1.2. Synthesis of 1-[2-[[5-(2-furanyl)-4-substituted
4H-1,2,4-triazol-3-yl]thio]ethyl]-2-methyl-5-
nitro-1H-imidazoles (3a–f)
To a solution/suspension of 2 (0.005 mol) in
CH3COCH3 (30 ml), 1 (0.005 mol) and K2CO3 (0.02
mol) were added. The reaction mixture was refluxed for
21 h, cooled and poured into ice water. The precipitate
was collected by filtration. Compounds 3a, 3b and 3d
were purified by washing with H2O and 3c, 3e and 3f
were recrystallized from C2H5OH.
Spectral data for 3a. IR [w cm−1, KBr]: 3107 (ꢁCꢀH),
1610 (CꢁN/CꢁC), 1522 (NO2), 1481, 1460, 1430 (CꢁN/
CꢁC), 1364 (NO2). 1H NMR [200 MHz, l ppm,
DMSO-d6]: 2.47 (s, 3H, CH3), 3.56–3.68 (m, 5H,
NꢀCH3 and SꢀCH2), 4.67 (t, 3H, NꢀCH2), 6.47 (dd,
J=3.0, 1.6 Hz, 1H, furan C4ꢀH), 7.10 (d, J=3.4
Hz,1H, furan C3ꢀH), 7.94–7.99 (m, 2H, furan C5ꢀH
and imidazole C4ꢀH). EIMS [m/z (%)]: 334 (M+, 2),
319 (5), 288 (100), 220 (7), 208 (27), 181(57), 154 (12),
139 (8), 121 (7), 108 (88), 94 (30), 80 (22), 67 (16).
Spectral data for 3d. IR [w cm−1, KBr]: 3124 (ꢁCꢀH),
1595 (CꢁN/CꢁC), 1524 (NO2), 1498, 1477, 1467 (CꢁN/
CꢁC), 1358 (NO2). 1H NMR [200 MHz, l ppm,
DMSO-d6]: 2.44 (s, 3H, CH3), 3.62 (t, J=6.3 Hz, 2H,
SꢀCH2), 4.70 (t, J=6.3 Hz, 2H, NꢀCH2), 6.15 (d,
J=3.2 Hz, 1H, furan C3ꢀH), 6.52 (dd, J=3.2, 1.6 Hz,
1H, furan C4ꢀH), 7.43–7.50 (m, 2H, ar.), 7.59–7.71 (m,
Spectral data for 5j. IR [w, cm−1, KBr]: 3230 (OꢀH),
3121 (ꢁCH), 1529 (NO2), 1514, 1467, 1427 (CꢁN/CꢁC),
1
1366 (NO2). H NMR [200 MHz, l ppm, DMSO-d6]:
2.43 (s, 3H, CH3), 2.44 (s, 3H, CH3), 3.39 (dd, J=5.8,
2.1 Hz, 2H, SꢀCH2), 4.04–4.12 (m, 1H, CH), 4.23 (dd,
J=13.7, 9.2 Hz, 1H, NꢀCH2), 4.56 (dd, J=13.7, 2.5
Hz, 1H, NꢀCH2), 5.00 (d, J=5.4 Hz, 1H, OH), 6.15 (d,
J=3.4 Hz, 1H, furan C3ꢀH), 6.51 (dd, J=8.4, 1.8 Hz,
1H, furan C4ꢀH), 7.29 (d, J=8.4 Hz, 2H, ar.), 7.34 (d
J=8.4 Hz, 2H, ar.), 7.82 (d, J=1.2 Hz, 1H, furan
C5ꢀH), 8.01 (s, 1H, imidazole C4ꢀH).
Spectral data for 5l. IR [w, cm−1, KBr]: 3247 (OꢀH),
3110 (ꢁCH), 1570 (NO2), 1524, 1498, 1464, 1444 (CꢁN/
CꢁC), 1365 (NO2). 1H NMR [200 MHz, l ppm,
DMSO-d6]: 2.44 (s, 3H, CH3), 3.38 (dd, J=5.6, 1.7 Hz,
2H, SꢀCH2), 4.04–4.12 (m, 1H, CH), 4.23 (dd, J=
14.1, 9.4 Hz, 1H, NꢀCH2), 4.55 (dd, J=14.1, 2.7 Hz,

830
N.S. Gu¨nay et al. / Il Farmaco 54 (1999) 826–831
1H, NꢀCH2), 5.65 (s, 1H, OH), 6.72 (d, J=3.4 Hz, 1H,
thiophene C3ꢀH), 7.00 (dd, J=3.9, 3.8 Hz, 1H, thio-
phene C4ꢀH), 7.52–7.66 (m, 5H, ar.), 8.01 (s, 1H,
imidazole C4ꢀH), 8.31 (s, 1H, thiophene, C5ꢀH).
ml nutrient broth (NB, Diagnostic Pasteur) and incu-
bated for 3–5 days at 25°C. At the end of the incuba-
tion period these strains were transferred into
screwcapped bottles containing sterilized beads and
shaken for 4–5 min in a vortex (IKA-VF, Germany).
The suspensions of the cultures were adjusted to have
an absorbance degree of 0.6 at 450 nm in the spec-
trophotometer. Eight different dilutions of the test
compounds between 25 and 0.2 mg/ml were prepared in
microplates by serial dilutions from top to bottom.
Then all the wells except the 12th wells (positive con-
trol) were filled with 10 ml of the standardized strains.
These plates were incubated at 25°C for 5 or 6 days.
The minimum concentration at which no growth was
observed was taken as the MIC value.
4.1.4. Synthesis of 1-[3-[(N,N-disubstituted
thiocarbamoyl)thio]-2-hydroxypropyl]-2-
methyl-5-nitro-1H-imidazoles (7a–b)
To a solution of 6 (0.0055 mol) in CH3COCH3 (30
ml), (0.0055 mol) 4 was added and the mixture was
refluxed for 4 h. The solid was separated by filtration
and the filtrate was allowed to cool to room tempera-
ture. The precipitate formed after cooling was collected
by filtration and recrystallized from C2H5OH.
Spectral data for 7b. IR [w, cm−1, KBr]: 3446 (OH),
3176 (ꢁCH), 1600 (CꢁN/CꢁC), 1533 (NO2), 1471, 1431
1
(CꢁN/CꢁC), 1361 (NO2). H NMR [200 MHz, l ppm,
Acknowledgements
DMSO-d6]: 2.46 (s, 3H, CH3), 3.28–3.46 (m, CH2
piperazine and solvent H2O), 3.59 (dd, J=13.6, 4.8 Hz,
2H, SꢀCH2), 3.94–4.09 (m, 1H, CH), 4.25 (dd, J=
13.9, 8.9 Hz, 5H, piperazine NꢀCH2 and CH2), 4.54
(dd, J=13.9, 3.5 Hz, 1H, NꢀCH2), 5.56 (d, J=5.8 Hz,
1H, OH), 6.81 (t, J=7.2 Hz, 1H, ar.), 6.95 (d, J=8.1
Hz, 2H, ar.), 7.25 (d, J=7.3 Hz, 2H, ar.), 8.00 (s, 1H,
imidazole C4ꢀH). EIMS [m/z (%)]: 421 (M+, 10), 404
(32), 375 (7), 352 (4), 293 (5), 263 (4), 238 (25), 205 (41),
189 (10), 184 (5), 171 (7), 162 (50), 133 (20), 132 (98),
120 (100), 104 (52), 91 (52), 91 (24), 85 (14), 78 (34), 77
(47), 63 (32).
This work was supported by Istanbul University
Research Fund, Project Number 686/301194.
References
[1] M. Bock, Ergebnisse experimenteller Versuche mit 1-(Hydroxy-
aethyl)-2-methyl-5-nitroimidazol an Trichomonas 6aginalis und
Entamoeba histolytica, Arzneim. Forsch. Drug Res. 11 (1961)
587–590.
[2] M. Hoffer, E. Grunberg, Synthesis and antiprotozoal activity of
1-(3-chloro-2-hydroxypropyl)-substituted nitroimidazoles, J.
Med. Chem. 17 (1974) 1019–1020.
[3] S.C. Bhatia, V.D. Shanbhag, Electron-capture gas chromato-
graphic assays of 5-nitroimidazole class of antimicrobials in
blood, J. Chromatogr. 305 (1984) 325–334.
4.2. Microbiology
4.2.1. Antibacterial acti6ity [22]
The disk diffusion method was used for the prelimi-
nary antibacterial evaluation of 3, 5 and 7. The MICs
of 5, which showed inhibition in the preliminary tests,
were determined by the microbroth dilution technique
using Mueller–Hinton broth (Difco Laboratories, De-
troit, MI). Serial two-fold dilutions of the test com-
pounds (235–0.6 mg/ml and 250–0.4 mg/ml) in DMSO
were prepared. The inoculum was prepared in broth,
which had been kept at 37°C overnight, and was di-
luted with broth to give a final concentration of 105
cfu/ml in the test tray. The trays were covered to
prevent drying. After incubation at 37°C for 18–20 h
the trays were examined for growth. The lowest concen-
tration of the test compounds inhibiting visible growth
was taken as the MIC value.
[4] M.P. Hay, W.R. Wilson, J.W. Moselen, B.D. Palmer, W.A.
Denny, Hypoxia-selective antitumor agents. 8. Bis(nitroimida-
zolyl)alkanecarboxamides: a new class of hypoxia-selective cyto-
toxins and hypoxic cell radiosensitizers, J. Med. Chem. 37 (1994)
381–391.
[5] S.S. Papakonstantinou-Garoufalias, O.G. Todoulou, E.C. Filip-
patos, A.E. Papadaki-Valiraki, A. Chytiroglou-Lada, Synthesis,
antifungal activity and antibacterial evaluation of some 3-piper-
azinylmethyl-5-aryl-1H-1,2,4-triazoles, Arzneim. Forsch. Drug
Res. 48 (1998) 1019–1023.
[6] S. Jantova´, G. Greif, R. Pavloviova´, L. Cipa´k, Antibacterial
effects of some 1-substituted 1,2,4-triazoles, Folia Microbiol.
(Praha) 43 (1998) 75–78.
[7] B.S. Holla, M.K. Shivananda, M.S. Shenoy, G. Antony, Studies
on arylfuran derivatives. Part VII. Synthesis and characterization
of some Mannich bases carrying halophenylfuryl moieties as
promising antibacterial agents, Farmaco 53 (1998) 531–535.
[8] C.H. Fuchsmann, W.H. Meek, Bactericidal and fungicidal hy-
droxythioaromatic acids and their derivatives, US 3557174;
1971. [Chem. Abstracts, 74 87636 m (1971)].
4.2.2. Antifungal acti6ity [23]
All the compounds to be tested were dissolved in
DMSO at a concentration of 4000 mg/ml and the final
concentration was reduced to 200 mg/ml with sterile
distilled water. No effect of DMSO (5%) was observed
upon growth of dermatophytes.
8
[9] A. Gu¨rsoy, O. Ates, N. Karali, N. Cesur, M. Kiraz, Synthesis
and antifungal activity of new carbamodithioic acid esters
derived from 3-acetylcoumarin, Eur. J. Med. Chem. 31 (1996)
643–646.
[10] L. Zaprutko, U. Wrzeciono, M. Gajdzinsky, M. Bartkowiak, K.
Lutomsky, W. Michalska, K. Pietkiewicz, Azole 34. Mitteilung:
Uber Nitroimidazolderivate und ihre antibakterielle und fun-
gizide Wirkung, Pharmazie 47 (1992) 258–261.
The dermatophyte strains which were grown on slant
medium of Sabouraud (Difco) were transferred to 3.5
8

N.S. Gu¨nay et al. / Il Farmaco 54 (1999) 826–831
831
[11] S¸. Demirayak, N. Kiraz, Some nitroimidazole derivatives as
possible antibacterial agents, Farmaco 48 (1993) 443–446.
[17] D.H. Williams, I. Fleming, Spectroscopic Methods in Organic
Chemistry, McGraw-Hill, London, 1989, pp. 85–101.
[18] M.M.A. Hassan, A.I. Jado, EI-Dine El-Shazly, M. Bahaa, Spec-
trophotometric quantitation of ornidazole and its formulations,
Spectrosc. Lett. 22 (1989) 111–121.
[19] H. Llempen, E. Salwinska, J. Suwinski, W. Szczepankiewicz, E.
Dziwinski, Nitroimidazoles. Part XVIII. EI Mass Spectrometry
of 1-aryl-4-nitroimidazoles in standard conditions, Polish J.
Chem. 67 (1993) 1745–1753.
[20] A.J. Blackman, J.H. Bowie, Electron impact studies. LXIX.
Mass spectra of 1,2,4-triazoles. I. Methyl- and phenyl-1,2,4-tria-
zoline-5-thiones, Aust. J. Chem. 25 (1972) 335–343.
[21] A. Bernardini, P. Viallefont, J. Daunis, Determination of the
structure of halo-1,a,4-triazoles by mass spectrometry, J. Hetero-
cycl. Chem. 12 (1975) 655–659.
8
[12] G. C¸ apan, N. Ergenc¸, G. Otu¨k-Sanis, Synthesis and biological
evaluation of 4-alkyl/aryl-1-(2-furoyl)-3-thiosemicarbazides and
4-alkyl/aryl-2,4-dihydro-5-(2-furyl)-3H-1,2,4-triazole-3-thiones,
J. Fac. Pharm. Istanbul (1990–92) 26–28; 23–30.
8
[13] N. Ergenc¸, N. Ulusoy, G. C¸ apan, G. Otu¨k Sanis, M. Kiraz,
Synthesis and antimicrobial properties of new 4-(alkylidene/
arylidene)amino-5-(2-furanyl)-2,4-dihydro-3H-1,2,4-triazole-3-
thiones and 6-aryl-3-(2-furanyl)-7H-1,2,4-triazolo[3,4-b][1,3,4]-
thiadiazines, Arch. Pharm. Pharm. Med. Chem. 329 (1996) 427–
430.
[14] G. C¸ apan, N. Ergenc¸, S. Bu¨yu¨ktimkin, N. Yulug, Synthesis
characterization and biological evaluation of 2-methyl-3-(N-sub-
stituted thiocarbamoylthio)acetamido-4(3H)-quinazolinones, Sci.
Pharm. 61 (1993) 243–250.
[22] R.N. Jones, A.L. Bary, T.L. Gavon, J.A. Washington, in: A.
Balows, W.J. Hausler, H.J. Shadomy (Eds.), Manual of Clinical
Microbiology, ASM, Washington, DC, 1985, p. 972.
[23] T.C. Granade, W.M. Artis, Antimycotic susceptibility testing of
dermatophytes in microcultures with a standardized fragmented
mycelial inoculum, Antimicrob. Agents Chemother. 17 (1980)
725–729.
[15] E.B. Akerblom, D.E.S. Campbell, Nitrofuryltriazole derivatives
as potential urinary tract antibacterial agents, J. Med. Chem. 16
(1973) 312–319.
[16] V.B. Jigajinni, S.C. Bennur, R.S. Bennur, V.V. Badiger, Synthe-
sis of 1-(2-furoyl)-4-substituted thiosemicarbazides and their
transformation to certain 5-membered heterocycles, J. Karnatak
Univ. Sci. 20 (1975) 1–10.
.