An investigation of the biological effect of structural modifications of isothiosemicarbazones and their cyclic analogues

Article abstract of DOI:10.1016/S0014-827X(03)00154-X

Several arylideneisothiosemicarbazones and arylidenehydrazothiazoles have been synthesised to obtain new antimicrobial agents. Their activity against both bacteria and fungi has been tested and some interesting informations about their biological activity

Full text of DOI:10.1016/S0014-827X(03)00154-X

Il Farmaco 58 (2003) 951ꢀ959
/
www.elsevier.com/locate/farmac
An investigation of the biological effect of structural modifications of
isothiosemicarbazones and their cyclic analogues
E. Maccioni a,*, M.C. Cardia a, S. Distinto a, L. Bonsignore a, A. De Logu b
a Dipartimento Farmaco Chimico Tecnologico, Universita degli Studi di Cagliari, Via Ospedale, 72, 09124 Cagliari, Italy
b Dipartimento di Scienze Chirurgiche e Trapianti d’Organo, Via Palabanda, 14, 09123 Cagliari, Italy
Received 5 December 2002; accepted 30 March 2003
Abstract
Several arylideneisothiosemicarbazones and arylidenehydrazothiazoles have been synthesised to obtain new antimicrobial agents.
Their activity against both bacteria and fungi has been tested and some interesting informations about their biological activity have
been obtained.
´
# 2003 Editions scientifiques et me´dicales Elsevier SAS. All rights reserved.
Keywords: Arylideneisothiosemicarbazones; Arylidenehydrazothiazoles; Anti-fungal and anti-microbial activity
1. Introduction
several species of Candida (C. albicans and C. krusei)
with MIC values comparable to those exhibited by
miconazole and clortrimazole.
To obtain further information on the influence of
structural modification on the biological activity, we
synthesised and tested a new series of 2-arylidency-
In previous communications we reported the synthesis
and antimicrobial activity of new arylideneisothiosemi-
carbazones. In particular we investigated the influence
of structural modification, such as the introduction of
substituents on the sulfur atom, the aromatic ring, or the
terminal nitrogen atom, on their biological activity
(Compounds A in Scheme 1) [1].
Most of the synthesised compounds show interesting
MIC values against some strains of Staphylococcus,
Bacillus, and Streptococcus, while none was active
against fungi.
cloalkylisothiosemicarbazones (3ꢀ
/
27), 2-hydrazothia-
62).
zoles (33ꢀ44), and arylidenehydrazothiazoles (45ꢀ
/
/
The group on the sulfur atom is either a mono- or di-
substituted benzyl, these being the most effective sub-
stitutions in the previously tested compounds [1,2].
More recently we synthesised and studied some
isothiosemicarbazones [2], where the arylidene group
was replaced with a cycloalkyl and the sulfur atom
either differently substituted or enclosed in a thiazole
ring (Compounds B and C in Scheme 1).
In most cases the cycloalkylisothiosemicarbazones (B)
were inactive against the tested bacteria and fungi,
except S. agalactiae. Interestingly we observed that the
higher the lipophilicity, the higher the activity.
On the contrary the corresponding cyclic analogues
(cycloalkyl hydrazo-thiazoles (C)) are active against
2. Results and discussion
2.1. Chemistry
2-Benzalcyclopentyliden-isothiosemicarbazones (3ꢀ
/
27) (Scheme 2) were synthesised by reacting isothiose-
micarbazides with the appropriate 2-arylidencyclopen-
tanone in refluxing isopropyl alcohol and a catalytic
amount of acetic acid.
The starting cyclopentanones were synthesised either
by reacting aldehydes with ketones in basic media or via
enamines as described in the literature.
In the case of compounds 28ꢀ
hydes are directly reacted with thiosemicarbazide, and
/
62, ketones or alde-
* Corresponding author.
E-mail address: maccione@unica.it (E. Maccioni).
´
0014-827X/03/$ - see front matter # 2003 Editions scientifiques et me´dicales Elsevier SAS. All rights reserved.
doi:10.1016/S0014-827X(03)00154-X

952
E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ959
/
Also in this case, isopropyl alcohol proved to be the
best solvent for our purpose. In fact, the reaction
products precipitate on cooling down, and can be
filtered and purified by crystallisation from ethanol or
ethanol/isopropanol.
All the synthesised products have been characterised
by analytical methods, Infrared (IR), 1H NMR, and
Mass Spectrometry.
2.2. Microbiology
All the synthesised compounds were tested against
several microbial species to investigate the influence of
structural modifications on biological activity.
In particular the antibacterial activity of compounds
3ꢀ62 was tested against five Gram-positive species
/
Scheme 1.
(Staphylococcus aureus, S. epidermidis, Streptococcus
agalactiae, S. faecalis, and B. subtilis), and five Gram-
negative species (Escherichia coli, Pseudomonas aerugi-
the obtained thiosemicarbazones reacted with a-halo-
genoketones to yield the 4-substituted thiazole ring.
Scheme 2.

E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ
/959
953
nosa, Salmonella typhi, Proteus mirabilis, and Klebsiella
pneumoniae), while their antifungal activity was evalu-
ated against C. albicans, C. glabrata, C. krusei, C.
parapsilosis, Saccharomyces cerevisiae, and C. tropica-
lis. All the microorganisms were isolated from clinical
specimens.
Nevertheless, interesting data can be obtained. In
particular, in the case of arylideneisothiosemicarba-
zones, the reduction in molecular flexibility and the
introduction of the arylidene moiety led to an almost
complete loss of antibacterial activity, while, in the case
of thiazole derivatives the antifungal activity is still
present in some of the tested compounds, meaning that
most probably the 2-hydrazo-4-substituted thiazole is
the active part of the molecule. However, the introduc-
tion of substituents with high steric hindrance reduces
biological activity.
In the case of compounds 3ꢀ27 a good antibacterial
/
activity was expected. As a matter of fact, in a previous
study on cycloalkylisothiosemicarbazones, we observed
that the higher the lipophilicity, the lower the MICs [2].
Compounds 3ꢀ27 have a higher lipophilic character
/
than the previously synthesised derivatives, due to the
introduction of the arylidene group on the cycloalkane
ring. For this reason an increase in antibacterial activity
was predictable.
4. Experimental
Unfortunately a dramatic decrease in activity was
observed, the MICs being higher than 200 mg/ml for all
the tested microorganisms. This suggests that though
important lipophilicity is not the only parameter that
4.1. Materials and methods
Melting points are uncorrected and were determined
on a Reichert Kofler thermopan apparatus. IR spectra
plays a key role in the structureꢀ
/
activity relationships of
these derivatives.
were recorded on a PerkinꢀElmer 1640 FT spectrometer
/
Other factors deriving from the introduction of the
arylidene group, such as an increase in both molecular
weight and conformational constriction, can seriously
affect the biological activity of these molecules.
Compounds 28ꢀ32 are intermediates and have not
been tested for antimicrobial activity.
As well as the previously synthesised thiazole deriva-
(KBr discs, in 1/cm). 1H NMR spectra were recorded on
a Bruker AMX (300 MHz) using tetramethylsilane
(TMS) as internal standard (chemical shifts in d values).
Electron ionisation (EI) mass spectra were obtained by a
Fisons QMD 1000 mass spectrometer (70 eV, 200 mA,
ion source temperature 200 8C). The samples were
introduced directly into the ion source. Elemental
/
tives [2], compounds 33ꢀ/62 were not active against the
analyses were obtained on a PerkinꢀElmer 240 B
/
tested bacteria, but in contrast with previous observa-
tions, almost no activity against fungi was measured.
Only compounds 34 and 44 exhibited antifungal
activity, but only at relatively high concentrations.
As a matter of fact, their MIC against C. albicans was
25 mg/ml, while Miconazole and Clotrimazole, used as
reference compounds, each exhibit a MIC of 3.12 mg/ml.
microanalyser.
4.2. Chemistry
The structures of all compounds were assigned on the
basis of IR, NMR, Mass spectra, and elemental
analysis.
Analytical data of the synthesised compounds are
reported in Tables 1ꢀ3.
In compounds 45ꢀ62 the cycloalkane ring was
/
eliminated and the arylidene group was directly con-
densed with the hydrazo-thiazole moiety to obtain more
active compounds.
/
Also in this case unfortunately the activity was lower
than expected, not only against bacteria but also against
fungi.
4.3. Synthesis of starting enamines, arylidencycloketones
and isothiosemicarbazides
Once more the presence of the planar aromatic ring,
with or without interposition of a cycloalkane ring, lead
to a complete loss both of antibacterial and antifungal
activity.
1-N-morpholinocyclopent-1-ene [3], 2-benzylidency-
clopentanone [4ꢀ6], and isothiosemicarbazides were
/
synthesised according to previously reported methods
[2]: 1 equiv. of thiosemicarbazide is refluxed in isopropyl
alcohol with 1 equiv. of the appropriate benzyl chloride.
The mixture is refluxed until complete dissolution (30ꢀ
/
3. Conclusions
90 min) and then allowed to cool down to r.t. The
required isothiosemicarbazide precipitates. After filtra-
tion, the product can be used without further purifica-
tion, or crystallised for characterisation with analytical
and spectroscopic methods.
Several arylideneisothiosemicarbazones and arylide-
nehydrazothiazoles were synthesised.
Unfortunately, none of the synthesised compounds
exhibited interesting biological properties, in particular
compared to analogous derivatives.
General method for the synthesis of compounds 3ꢀ
/
27.

954
E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ959
/
Table 1
2-Benzalcyclopentyliden-S-benzylisothiosemicarbazones
Comp.
Formula
R
X
m/z
Yield %
M.p., 8C
C, %
H, %
N, %
3
C
20H20N3ClS
C20H20N3ClS
20H19N3Cl2S
C20H20N3Cl2S
22H25N3S
C20H18N4Cl2O2S
20H18N4Cl2O2S
C20H18N3Cl3S
20H17N3Cl4S
H
2-Cl
4-Cl
369
369
404
404
363
449
449
438
479
438
449
449
438
479
414
414
414
404
438
408
408
414
404
397
432
89
88
95
83
81
90
88
92
95
95
88
92
95
87
93
87
85
90
95
85
81
86
92
87
89
218ꢀ
221
206ꢀ
218ꢀ
206ꢀ
/
219
64.94 (65.05)
64.94 (64.71)
59.41 (59.37)
59.41 (59.35)
72.69 (72.55)
53.46 (53.29)
53.46 (53.32)
54.74 (54.92)
50.76 (50.61)
54.74 (54.89)
53.46 (53.61)
53.46 (53.64)
54.74 (54.60)
50.76 (50.86)
57.90 (57.73)
57.90 (58.00)
57.90 (57.72)
59.41 (59.55)
54.74 (54.63)
64.68 (64.75)
64.68 (64.80)
57.90 (58.07)
59.41 (59.18)
66.40 (66.29)
61.11 (61.27)
5.45 (5.48)
5.45 (5.41)
4.74 (4.71)
4.74 (4.77)
6.93 (6.90)
4.04 (4.01)
4.04 (4.07)
4.13 (4.16)
3.58 (3.61)
4.13 (4.11)
4.04 (4.01)
4.04 (4.07)
4.13 (4.15)
3.58 (3.61)
4.62 (4.64)
4.62 (4.59)
4.62 (4.59)
4.74 (4.76)
4.13 (4.10)
5.92 (5.96)
5.92 (5.90)
4.62 (4.66)
4.74 (4.72)
6.08 (6.10)
5.36 (5.38)
11.36 (11.43)
11.36 (11.31)
10.39 (10.35)
10.39 (10.42)
11.55 (11.60)
12.47 (12.45)
12.47 (12.50)
9.58 (9.61)
4
H
5
C
H
3,4-Cl
2,4-Cl
3,4-CH3
3,4-Cl
3,4-Cl
3,4-Cl
3,4-Cl
4-Cl
/
210
219
208
220
6
H
/
7
C
H
/
8
4-NO2
3-NO2
4-Cl
218ꢀ
/
9
C
225
214
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
C
2,4-Cl
2,4-Cl
4-NO2
3-NO2
4-Cl
206ꢀ
204ꢀ
221ꢀ
228ꢀ
211ꢀ
218ꢀ
220ꢀ
229ꢀ
/
210
206
222
229
213
219
221
231
221
8.77 (8.80)
9.58 (9.62)
C20H18N3Cl3S
/
C20H18N4Cl2O2S
C20H18N4Cl2O2S
2,4-Cl
2,4-Cl
2,4-Cl
2,4-Cl
4-Cl
/
12.47 (12.50)
12.47 (12.43)
9.58 (9.54)
/
C
20H18N3Cl3S
/
C20H17N3Cl4S
C20H19N4ClO2S
2,4-Cl
4-NO2
4-NO2
3-NO2
4-Cl
/
8.77 (8.83)
/
13.50 (13.47)
13.50 (13.45)
13.50 (13.48)
10.39 (10.36)
9.58 (9.62)
C
20H19N4ClO2S
C20H19N4ClO2S
20H19N3Cl2S
C20H18N3Cl3S
22H24N4O2S
2-Cl
2-Cl
/
220ꢀ
/
C
2-Cl
232
2,4-Cl
4-NO2
3-NO2
3-NO2
4-Cl
2-Cl
223ꢀ
222ꢀ
233ꢀ
/
226
224
235
221
C
3,4-CH3
3,4-CH3
4-Cl
/
13.72 (13.69)
13.72 (13.78)
13.50 (13.56)
10.39 (10.44)
10.56 (10.52)
9.72 (9.69)
C22H24N4O2S
C20H19N4ClO2S
C20H19N3Cl2S
/
218ꢀ
/
4-Cl
217
C22H24N3ClS
C22H23N3Cl2S
4-Cl
2,4-Cl
3,4-CH3
3,4-CH3
216ꢀ
/
218
210
4.4. Synthesis of 2-benzalcyclopentyliden-S-(2-
chlorobenzyl)-isothiosemicarbazone (3)
1H NMR (DMSO): d 1.91ꢀ
2.90 (m, 4H, CH2); 4.86 (s, 2H, CH2Ã
); 10.05 (s, 1H, NH2, D-
/
2.00 (m, 2H, CH2); 2.47ꢀ
/
/
S); 7.39ꢀ8.42 (m,
/
10H, phenyl-H and phenyl-CHÄ
/
exch.); 12.63 (s, 1H, NH2, D-exch.).
According to the same procedure, the following listed
compounds have been synthesised.
In a flask equipped with a refluxing condenser and a
magnetic stirrer, 2-benzalcyclopentanone (5 mmol) and
isothiosemicarbazide are reacted in 60 ml of isopropyl
alcohol and a catalytic amount of acetic aid. The
mixture is stirred at room temperature for 5ꢀ10 min,
/
and then refluxed to complete dissolution. The solution
is further refluxed for 20 min. On cooling, a solid is
formed, which can be filtered, washed with water, and
crystallised from ethanol.
4.4.1. 2-Benzalcyclopentyliden-S-(4-chlorobenzyl)-
isothiosemicarbazone (4)
1H NMR (DMSO): d 1.95ꢀ
/
2.00 (m, 2H, CH2); 2.46ꢀ
/
2.90 (m, 4H, CH2); 4.88 (s, 2H, CH2ÃS); 7.36ꢀ8.40 (m,
/
/
Table 2
Thiazoles-2,4-substituted 33ꢀ44
/
Comp.
Formula
R
R?
m/z
M.p., 8C
C, %
H, %
N, %
33
34
35
36
37
38
39
40
41
42
43
44
C
21H19N3S
C16H17N3S
H
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
CH3
C6H5
345
283
390
328
390
328
414
352
379
317
241
303
228ꢀ
219ꢀ
260ꢀ
217ꢀ
249ꢀ
218ꢀ
190ꢀ
206ꢀ
230ꢀ
206ꢀ
247ꢀ
220ꢀ
/
230
220
262
218
250
220
191
207
231
208
248
223
73.02 (72.75)
67.81 (68.03)
64.60 (64.32)
58.52 (58.70)
64.60 (64.91)
58.52 (58.72)
60.87 (61.03)
54.55 (54.36)
66.39 (66.15)
60.46 (60.72)
49.77 (50.01)
59.39 (59.59)
5.54 (5.57)
6.05 (6.02)
4.65 (4.68)
4.91 (4.89)
4.65 (4.62)
4.91 (4.94)
4.14 (4.11)
4.29 (4.32)
4.77 (4.79)
5.07 (5.04)
6.27 (6.31)
5.65 (5.68)
12.16 (12.20)
14.83 (14.77)
14.35 (14.40)
17.06 (16.99)
14.35 (14.29)
17.06 (17.10)
10.14 (10.10)
11.93 (11.98)
11.06 (11.08)
13.22 (13.16)
17.41 (17.16)
13.85 (13.91)
H
/
C21H18N4O2S
C16H16N4O2S
4-NO2
4-NO2
3-NO2
3-NO2
2,4-Cl
2,4-Cl
4-Cl
/
/
C
21H18N4O2S
/
C16H16N4O2S
C21H17N3Cl2S
C16H15N3Cl2S
/
/
/
C
21H18N3ClS
/
C16H16N3ClS
C10H15N3O2S
C15H17N3O2S
4-Cl
/
ꢀ/
ꢀ/
/
/

E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ
/959
955
Table 3
Thiazoles 3,4-disobstituted 45ꢀ
/62
Comp.
Formula
R
R1
m/z
M.p., 8C
C, %
H, %
N, %
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
C
11H11N3S
C16H13N3S
11H10N3ClS
C16H12N3ClS
11H9N3Cl2S
C16H11N3Cl2S
11H10N4O2S
C16H12N4O2S
11H10N4O2S
H
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
CH3
C6H5
217
279
251
313
286
348
262
324
262
324
262
324
247
309
277
339
231
293
192ꢀ
252ꢀ
207ꢀ
246ꢀ
224ꢀ
244ꢀ
215ꢀ
263ꢀ
205ꢀ
241ꢀ
231ꢀ
245
199ꢀ
213ꢀ
223ꢀ
236ꢀ
181ꢀ
202ꢀ
/
194
255
209
248
226
246
217
265
206
244
232
60.80 (61.02)
68.79 (68.58)
52.49 (52.55)
61.24 (61.07)
46.17 (45.98)
55.18 (54.98)
50.37 (50.26)
59.25 (59.13)
50.37 (50.29)
59.25 (59.34)
50.37 (50.30)
59.25 (59.29)
58.28 (58.19)
66.00 (65.95)
56.30 (56.22)
63.70 (63.63)
62.31 (62.19)
69.60 (69.48)
5.10 (5.14)
4.69 (4.73)
4.00 (4.03)
3.86 (3.84)
3.17 (3.19)
3.18 (3.20)
3.84 (3.87)
3.73 (3.70)
3.84 (3.82)
3.73 (3.71)
3.84 (3.81)
3.73 (3.75)
5.30 (5.27)
4.89 (4.93)
5.45 (5.43)
5.05 (5.08)
5.66 (5.63)
5.15 (5.12)
19.34 (19.28)
15.04 (14.99)
16.70 (16.65)
13.39 (13.44)
14.68 (14.64)
12.07 (12.04)
21.36 (21.31)
17.27 (17.23)
21.36 (21.30)
17.27 (17.29)
21.36 (21.41)
17.27 (17.30)
16.99 (17.04)
13.58 (13.62)
15.15 (15.09)
12.38 (12.34)
18.17 (18.12)
14.32 (14.37)
H
/
C
4-Cl
/
4-Cl
/
C
2,4-Cl
2,4-Cl
2-NO2
2-NO2
3-NO2
3-NO2
4-NO2
4-NO2
4-OCH3
4-OCH3
3,4-OCH3
3,4-OCH3
4-CH3
4-CH3
/
/
C
/
/
C
/
C16H12N4O2S
C11H10N4O2S
C16H12N4O2S
/
/
C
12H13N3OS
/
203
217
226
238
184
205
C17H15N3OS
C13H15N3O2S
/
/
C
18H17N3O2S
C12H13N3S
17H15N3S
/
/
C
/
10H, phenyl-H and phenyl-CHÄ
/
); 10.00 (s, 1H, NH2, D-
4.4.6. 2-(3-Nitrobenzalcyclopentyliden)-S-(3,4-
dichlorobenzyl)-isothiosemicarbazone (9)
exch.); 12.60 (s, 1H, NH2, D-exch.).
1H NMR (DMSO): d 1.87ꢀ
/
1.95 (m, 2H, CH2); 2.67ꢀ
2.86 (m, 4H, CH2); 4.68 (s, 2H, CH2ÃS); 7.50ꢀ8.27 (m,
); 9.95 (s, 1H, NH2, D-
/
/
/
4.4.2. 2-Benzalcyclopentyliden-S-(3,4-dichlorobenzyl)-
isothiosemicarbazone (5)
8H, phenyl-H and phenyl-CHÄ
/
exch.); 12.65 (s, 1H, NH2, D-exch.).
1H NMR (DMSO): d 1.99ꢀ
/
2.07 (m, 2H, CH2); 2.48ꢀ
2.95 (m, 4H, CH2); 4.87 (s, 2H, CH2ÃS); 7.32ꢀ8.39 (m,
); 10.03 (s, 1H, NH2, D-
/
/
/
4.4.7. 2-(4-Chlorobenzalcyclopentyliden)-S-(3,4-
9H, phenyl-H and phenyl-CHÄ
/
dichlorobenzyl)-isothiosemicarbazone (10)
exch.); 12.69 (s, 1H, NH2, D-exch.).
1H NMR (DMSO): d 1.84ꢀ
/
2.20 (m, 2H, CH2); 2.51ꢀ
2.99 (m, 4H, CH2); 4.78 (s, 2H, CH2ÃS); 7.35ꢀ7.99 (m,
); 9.96 (s, 1H, NH2, D-
/
/
/
8H, phenyl-H and phenyl-CHÄ
/
4.4.3. 2-Benzalcyclopentyliden-S-(2,4-dichlorobenzyl)-
isothiosemicarbazone (6)
exch.); 12.58 (s, 1H, NH2, D-exch.).
1H NMR (DMSO): d 1.94ꢀ
/
1.99 (m, 2H, CH2); 2.60ꢀ
2.95 (m, 4H, CH2); 4.75 (s, 2H, CH2ÃS); 7.36ꢀ8.33 (m,
9H, phenyl-H and phenyl-CHÄ); 10.25 (s, 1H, NH2);
12.58 (s, 1H, NH2).
/
4.4.8. 2-(2,4-Dichlorobenzalcyclopentyliden)-S-(3,4-
dichlorobenzyl)-isothiosemicarbazone (11)
/
/
/
1H NMR (DMSO): d 1.90ꢀ
/
1.95 (m, 2H, CH2); 2.60ꢀ
2.85 (m, 4H, CH2); 4.74 (s, 2H, CH2ÃS); 7.45ꢀ7.92 (m,
); 9.79 (s, 1H, NH2, D-
/
/
/
7H, phenyl-H and phenyl-CHÄ
/
exch.); 12.52 (s, 1H, NH2, D-exch.).
4.4.4. 2-Benzalcyclopentyliden-S-(3,4-dimethylbenzyl)-
isothiosemicarbazone (7)
4.4.9. 2-(2,4-Dichlorobenzalcyclopentyliden)-S-(4-
chlorobenzyl)-isothiosemicarbazone (12)
1H NMR (CDCl3): d 1.88ꢀ
/
1.99 (m, 2H, CH2); 2.22ꢀ
2.85 (m, 10H, CH2 and phenyl-CH3); 4.53 (s, 2H, CH2Ã
S); 7.12ꢀ ); 9.95
/
/
1H NMR (DMSO): d 1.89ꢀ
/
2.02 (m, 2H, CH2); 2.56ꢀ
2.98 (m, 4H, CH2); 4.77 (s, 2H, CH2ÃS); 7.44ꢀ8.00 (m,
); 9.97 (s, 1H, NH2, D-
/
/
7.45 (m, 9H, phenyl-H and phenyl-CHÄ
/
/
/
(s, 1H, NH2, D-exch.); 12.42 (s, 1H, NH2, D-exch.).
8H, phenyl-H and phenyl-CHÄ
/
exch.); 12.57 (s, 1H, NH2, D-exch.).
4.4.5. 2-(4-Nitrobenzalcyclopentyliden)-S-(3,4-
dichlorobenzyl)-isothiosemicarbazone (8)
4.4.10. 2-(4-Nitrobenzalcyclopentyliden)-S-(2,4-
dichlorobenzyl)-isothiosemicarbazone (13)
1H NMR (DMSO): d 1.96ꢀ
/
2.10 (m, 2H, CH2); 2.40ꢀ
/
1H NMR (DMSO): d 1.98ꢀ
/
2.17 (m, 2H, CH2); 2.22ꢀ
2.97 (m, 4H, CH2); 4.89 (s, 2H, CH2ÃS); 7.42ꢀ8.40 (m,
8H, phenyl-H and phenyl-CHÄ); 10.04 (s, 1H, NH2, D-
exch.); 12.69 (s, 1H, NH2, D-exch.).
/
3.05 (m, 4H, CH2); 4.86 (s, 2H, CH2Ã
8H, phenyl-H and phenyl-CHÄ
exch.); 12.76 (s, 1H, NH2, D-exch.).
/
S); 7.35ꢀ
/
8.40 (m,
/
/
/
); 10.10 (s, 1H, NH2, D-
/

956
E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ959
/
4.4.11. 2-(3-Nitrobenzalcyclopentyliden)-S-(2,4-
dichlorobenzyl)-isothiosemicarbazone (14)
4.4.19. 2-(4-Nitrobenzalcyclopentyliden)-S-(3,4-
dimethylbenzyl)-isothiosemicarbazone (22)
1H NMR (DMSO): d 1.99ꢀ
/
2.04 (m, 2H, CH2); 2.38ꢀ
2.99 (m, 4H, CH2); 4.74 (s, 2H, CH2ÃS); 7.50ꢀ8.40 (m,
); 9.69 (s, 1H, NH2, D-
/
1H NMR (DMSO): d 1.90ꢀ
/
2.85 (m, 12H, CH2 and
phenyl-CH3); 4.70 (s, 2H, CH2ÃS); 7.21ꢀ8.30 (m, 8H,
); 10.10 (s, 1H, NH2, D-
/
/
/
/
8H, phenyl-H and phenyl-CHÄ
/
phenyl-H and phenyl-CHÄ
/
exch.); 12.61 (s, 1H, NH2, D-exch.).
exch.); 12.77 (s, 1H, NH2, D-exch.).
4.4.12. 2-(4-Chlorobenzalcyclopentyliden)-S-(2,4-
dichlorobenzyl)-isothiosemicarbazone (15)
4.4.20. 2-(3-Nitrobenzalcyclopentyliden)-S-(3,4-
dimethylbenzyl)-isothiosemicarbazone (23)
1H NMR (DMSO): d 1.93ꢀ
/
2.05 (m, 2H, CH2); 2.50ꢀ
3.20 (m, 4H, CH2); 4.78 (s, 2H, CH2ÃS); 7.50ꢀ7.60 (m,
); 9.83 (s, 1H, NH2, D-
/
1H NMR (DMSO): d 2.01ꢀ
/
2.04 (m, 2H, CH2); 2.27ꢀ
2.96 (m, 10H, CH2 and phenyl-CH3); 4.63 (s, 2H, CH2Ã
S); 7.20ꢀ ); 9.79
/
/
/
/
8H, phenyl-H and phenyl-CHÄ
/
/
8.38 (m, 8H, phenyl-H and phenyl-CHÄ
/
exch.); 12.48 (s, 1H, NH2, D-exch.).
(s, 1H, NH2, D-exch.); 12.71 (s, 1H, NH2, D-exch.).
4.4.13. 2-(2,4-Dichlorobenzalcyclopentyliden)-S-(2,4-
dichlorobenzyl)-isothiosemicarbazone (16)
4.4.21. 2-(3-Nitrobenzalcyclopentyliden)-S-(4-
chlorobenzyl)-isothiosemicarbazone (24)
1H NMR (DMSO): d 1.60ꢀ
/
2.20 (m, 2H, CH2); 2.45ꢀ
3.20 (m, 4H, CH2); 4.79 (s, 2H, CH2ÃS); 7.30ꢀ8.35 (m,
); 10.30 (s, 1H, NH2, D-
/
1H NMR (DMSO): d 1.90ꢀ
/
1.93 (m, 2H, CH2); 2.66ꢀ
/
/
/
2.85 (m, 4H, CH2); 4.62 (s, 2H, CH2Ã
/
S); 7.40ꢀ
/
8.27 (m,
); 9.68 (s, 1H, NH2, D-
7H, phenyl-H and phenyl-CHÄ
/
9H, phenyl-H and phenyl-CHÄ
/
exch.); 12.60 (s, 1H, NH2, D-exch.).
exch.); 12.48 (s, 1H, NH2, D-exch.).
4.4.14. 2-(4-Nitrobenzalcyclopentyliden)-S-(4-
chlorobenzyl)-isothiosemicarbazone (17)
4.4.22. 2-(4-Chlorobenzalcyclopentyliden)-S-(4-
chlorobenzyl)-isothiosemicarbazone (25)
1H NMR (DMSO): d 1.88ꢀ
/
1.93 (m, 2H, CH2); 2.88ꢀ
3.27 (m, 4H, CH2); 4.88 (s, 2H, CH2ÃS); 7.33ꢀ8.37 (m,
); 9.65 (s, 1H, NH2, D-
/
1H NMR (DMSO): d 1.89ꢀ
/
2.03 (m, 2H, CH2); 2.37ꢀ
2.84 (m, 4H, CH2); 4.70 (s, 2H, CH2ÃS); 7.52ꢀ8.33 (m,
); 9.77 (s, 1H, NH2, D-
/
/
/
/
/
9H, phenyl-H and phenyl-CHÄ
/
9H, phenyl-H and phenyl-CHÄ
/
exch.); 12.85 (s, 1H, NH2, D-exch.).
exch.); 12.42 (s, 1H, NH2, D-exch.).
4.4.15. 2-(4-Nitrobenzalcyclopentyliden)-S-(2-
chlorobenzyl)-isothiosemicarbazone (18)
4.4.23. 2-(4-Chlorobenzalcyclopentyliden)-S-(3,4-
dimethylbenzyl)-isothiosemicarbazone (26)
1H NMR (DMSO): d 1.92ꢀ
/
2.03 (m, 2H, CH2); 2.40ꢀ
3.09 (m, 4H, CH2); 4.79 (s, 2H, CH2ÃS); 7.35ꢀ8.46 (m,
); 10.01 (s, 1H, NH2, D-
/
1H NMR (DMSO): d 1.96ꢀ
/
2.00 (m, 2H, CH2); 2.20ꢀ
2.88 (m, 10H, CH2 and phenyl-CH3); 4.79 (s, 2H, CH2Ã
S); 7.23ꢀ ); 9.80
/
/
/
/
9H, phenyl-H and phenyl-CHÄ
/
/
7.95 (m, 8H, phenyl-H and phenyl-CHÄ
/
exch.); 12.68 (s, 1H, NH2, D-exch.).
(s, 1H, NH2, D-exch.); 12.44 (s, 1H, NH2, D-exch.).
4.4.16. 2-(3-Nitrobenzalcyclopentyliden)-S-(2-
chlorobenzyl)-isothiosemicarbazone (19)
4.4.24. 2-(2,4-Dichlorobenzalcyclopentyliden)-S-(3,4-
dimethylbenzyl)-isothiosemicarbazone (27)
1H NMR (DMSO): d 1.87ꢀ
/
1.92 (m, 2H, CH2); 2.40ꢀ
2.85 (m, 4H, CH2); 4.64 (s, 2H, CH2ÃS); 7.43ꢀ8.30 (m,
); 10.00 (s, 1H, NH2, D-
/
1H NMR (DMSO): d 1.97ꢀ
/
2.05 (m, 2H, CH2); 2.13ꢀ
2.91 (m, 10H, CH2 and phenyl-CH3); 4.70 (s, 2H, CH2Ã
S); 7.13ꢀ8.35 (m, 7H, phenyl-H and phenyl-CHÄ); 9.82
(s, 1H, NH2, D-exch.); 12.73 (s, 1H, NH2, D-exch.).
General method for the synthesis of compounds 33ꢀ
62.
/
/
/
/
9H, phenyl-H and phenyl-CHÄ
/
/
/
exch.); 12.77 (s, 1H, NH2, D-exch.).
/
4.4.17. 2-(4-Chlorobenzalcyclopentyliden)-S-(2-
chlorobenzyl)-isothiosemicarbazone (20)
1H NMR (DMSO): d 1.91ꢀ
/
2.01 (m, 2H, CH2); 2.39ꢀ
2.72 (m, 4H, CH2); 4.73 (s, 2H, CH2ÃS); 7.38ꢀ8.22 (m,
); 10.09 (s, 1H, NH2, D-
/
4.5. Synthesis of 2-[2-(benzalcyclopentyliden)hydrazo]-
4-phenylthiazole (33)
/
/
9H, phenyl-H and phenyl-CHÄ
/
exch.); 12.53 (s, 1H, NH2, D-exch.).
2-Benzalcyclopentylidenthiosemicarbazide (0.02 mol)
and v-bromo-acetophenone (0.02 mol) are stirred in
4.4.18. 2-(2,4-Dichlorobenzalcyclopentyliden)-S-(2-
chlorobenzyl)-isothiosemicarbazone (21)
refluxing isopropyl alcohol (70ꢀ100 ml) to complete
/
dissolution, and then until a white foaming product is
formed. The mixture is then allowed to cool down and
the solid is filtered, washed with saturated NaHCO3
water solution and then with cool water, dried, and
crystallised from ethanol.
1H NMR (DMSO): d 1.96ꢀ
/
2.03 (m, 2H, CH2); 2.43ꢀ
2.79 (m, 4H, CH2); 4.83 (s, 2H, CH2ÃS); 7.41ꢀ8.34 (m,
8H, phenyl-H and phenyl-CHÄ); 10.13 (s, 1H, NH2, D-
exch.); 12.64 (s, 1H, NH2, D-exch.).
/
/
/
/

E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ
/959
957
1H NMR (CDCl3): d 1.98ꢀ
2.95 (m, 4H, CH2); 6.74 (s, 1H, H5-thiaz.); 7.25ꢀ
11H, phenyl-H and phenyl-CHÄ); 12.18 (s, 1H, NH, D-
exch.).
According to this method, the following listed com-
pounds have been synthesised.
/
2.09 (m, 2H, CH2); 2.83ꢀ
/
4.5.8. 2-[2-(4-Chlorobenzalcyclopentyliden)hydrazo]-4-
phenylthiazole (41)
1H NMR (CDCl3): d 1.98ꢀ
2.88 (m, 4H, CH2); 6.74 (s, 1H, H5-thiaz.); 7.25ꢀ
10H, phenyl-H and phenyl-CHÄ); 12.47 (s, 1H, NH, D-
exch.).
/7.75 (m,
/
/
2.08 (m, 2H, CH2); 2.63ꢀ
/
/
8.00 (m,
/
4.5.9. 2-[2-(4-Chlorobenzalcyclopentyliden)hydrazo]-4-
methylthiazole (42)
4.5.1. 2-[2-(Benzalcyclopentyliden)hydrazo]-4-
methylthiazole (34)
1H NMR (CDCl3): d 1.94ꢀ
/
2.15 (m, 2H, CH2); 2.37 (s,
2.84 (m, 4H, CH2); 6.72 (s, 1H, H5-
);
1H NMR (CDCl3): d 2.02ꢀ
/
2.22 (m, 2H, CH2); 2.41 (s,
2.91 (m, 4H, CH2); 6.25 (s, 1H, H5-
);
3H, CH3); 2.62ꢀ
/
3H, CH3); 2.81ꢀ
/
thiaz.); 7.54ꢀ
/
7.85 (m, 5H, phenyl-H and phenyl-CHÄ
/
thiaz.); 7.32ꢀ
/
7.52 (m, 6H, phenyl-H and phenyl-CHÄ
/
13.28 (s, 1H, NH, D-exch.).
12.56 (s, 1H, NH, D-exch.).
4.5.10. 2-[1-(Carbetoxymethylethyliden)hydrazo]-4-
methylthiazole (43)
4.5.2. 2-[2-(4-Nitrobenzalcyclopentyliden)hydrazo]-4-
phenylthiazole (35)
1H NMR (CDCl3): d 1.16 (t, 3H, OCH2CH3); 2.29 (s,
1H NMR (CDCl3): d 1.98ꢀ
/
2.00 (m, 2H, CH2); 2.75ꢀ
2.80 (m, 4H, CH2); 6.64 (s, 1H, H5-thiaz.); 7.07ꢀ
8.14 (m,
10H, phenyl-H and phenyl-CHÄ); 12.47 (s, 1H, NH, D-
exch.).
/
3H, CH3); 2.35 (s, 3H, CH3); 3.54 (s, 2H, CH2CO);
3.96ꢀ
4.02 (q, 2H, OCH2); 6.70 (s, 1H, H5-thiaz.); 12.58
(s, 1H, NH, D-exch.).
/
/
/
4.5.11. 2-[1-(Carbetoxymethylethyliden)hydrazo]-4-
phenylthiazole (44)
4.5.3. 2-[2-(4-Nitrobenzalcyclopentyliden)hydrazo]-4-
methylthiazole (36)
1H NMR (CDCl3): d 1.29 (t, 3H, OCH2CH3); 2.29 (s,
1H NMR (CDCl3): d 1.99ꢀ
/
2.16 (m, 2H, CH2); 2.37 (s,
2.93 (m, 4H, CH2); 6.24 (s, 1H, H5-
);
3H, CH3); 3.41 (s, 2H, CH2CO); 4.17ꢀ
OCH2); 6.73 (s, 1H, H5-thiaz.); 7.25ꢀ
7.73 (m, 5H,
phenyl-H); 12.63 (s, 1H, NH, D-exch.).
/4.24 (q, 2H,
3H, CH3); 2.82ꢀ
/
/
thiaz.); 7.25ꢀ
/
8.25 (m, 5H, phenyl-H and phenyl-CHÄ
/
13.02 (s, 1H, NH, D-exch.).
4.5.12. 2-Benzylidenhydrazo-4-methylthiazole (45)
1H NMR (DMSO): d 2.27 (s, 3H, CH3); 6.74 (s, 1H,
4.5.4. 2-[2-(3-Nitrobenzalcyclopentyliden)hydrazo]-4-
phenylthiazole (37)
H5-thiaz); 7.45ꢀ
phenyl-CHÄ
/7.79 (m, 5H, phenyl-H); 8.47 (s, 1H,
1H NMR (CDCl3): d 2.03ꢀ
/
2.13 (m, 2H, CH2); 2.88ꢀ
2.95 (m, 4H, CH2); 6.77 (s, 1H, H5-thiaz.); 7.25ꢀ
8.31 (m,
10H, phenyl-H and phenyl-CHÄ); 12.64 (s, 1H, NH, D-
exch.).
/
/); 10.46 (s, 1H, NH, D-exch.).
/
/
4.5.13. 2-Benzylidenhydrazo-4-phenylthiazole (46)
1H NMR (CDCl3): d 6.77 (s, 1H, H5-thiaz.); 7.43ꢀ
7.74 (m, 10H, phenyl-H); 8.20 (s, 1H, phenyl-CHÄ);
10.39 (s, 1H, NH, D-exch.).
/
/
4.5.5. 2-[2-(3-Nitrobenzalcyclopentyliden)hydrazo]-4-
methylthiazole (38)
1H NMR (CDCl3): d 2.05ꢀ
/
2.15 (m, 2H, CH2); 2.43 (s,
2.99 (m, 4H, CH2); 6.30 (s, 1H, H5-
);
3H, CH3); 2.87ꢀ
/
4.5.14. 2-(4-Chlorobenzyliden)hydrazo-4-methylthiazole
(47)
thiaz.); 7.31ꢀ
/
8.35 (m, 5H, phenyl-H and phenyl-CHÄ
/
1H NMR (DMSO): d 2.27 (s, 3H, CH3); 6.72 (s, 1H,
12.99 (s, 1H, NH, D-exch.).
H5-thiaz.); 7.51ꢀ
phenyl-CHÄ
/
7.83 (m, 4H, phenyl-H); 8.40 (s, 1H,
/
); 11.02 (s, 1H, NH, D-exch.).
4.5.6. 2-[2-(2,4-Dichlorobenzalcyclopentyliden)-
hydrazo]-4-phenylthiazole (39)
1H NMR (CDCl3): d 2.02ꢀ
/
2.10 (m, 2H, CH2); 2.85ꢀ
2.93 (m, 4H, CH2); 6.72 (s, 1H, H5-thiaz.); 7.18ꢀ
7.55 (m,
9H, phenyl-H and phenyl-CHÄ); 12.70 (s, 1H, NH, D-
exch.).
/
4.5.15. 2-(4-Chlorobenzyliden)hydrazo-4-phenylthiazole
(48)
/
1H NMR (DMSO): d 7.40ꢀ
/
7.99 (m, 9H, phenyl-H);
/
7.46 (s, 1H, H5-thiaz.); 8.15 (s, 1H, phenyl-CHÄ
(s, 1H, NH, D-exch.).
/); 12.36
4.5.7. 2-[2-(2,4-Dichlorobenzalcyclopentyliden)-
hydrazo]-4-methylthiazole (40)
4.5.16. 2-(2,4-Dichlorobenzyliden)hydrazo-4-
methylthiazole (49)
1H NMR (CDCl3): d 1.90ꢀ
/
2.15 (m, 2H, CH2); 2.44 (s,
2.71 (m, 4H, CH2); 6.08 (s, 1H, H5-
7.42 (m, 4H, phenyl-H and phenyl-CHÄ);
12.85 (s, 1H, NH, D-exch.).
3H, CH3); 2.60ꢀ
/
1H NMR (DMSO): d 2.27 (s, 3H, CH3); 6.72 (s, 1H,
thiaz.); 7.10ꢀ
/
/
H5-thiaz.); 7.52ꢀ
phenyl-CHÄ); 11.85 (s, 1H, NH, D-exch.).
/8.07 (m, 3H, phenyl-H); 8.66 (s, 1H,
/

958
E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ959
/
4.5.17. 2-(2,4-Dichlorobenzyliden)hydrazo-4-
phenylthiazole (50)
4.5.26. 2-(3,4-Dimethoxybenzyliden)hydrazo-4-
methylthiazole (59)
1H NMR (DMSO): d 6.80 (s, 1H, H5-thiaz.); 7.42ꢀ
/
1H NMR (DMSO): d 2.32 (s, 3H, CH3); 3.85 (s, 6H,
8.05 (m, 8H, phenyl-H); 8.46 (s, 1H, phenyl-CHÄ
(s, 1H, NH, D-exch.).
/); 12.30
OCH3); 6.74 (s, 1H, H5-thiaz.); 7.05ꢀ
/
7.50 (m, 3H,
); 11.75 (s, 1H,
phenyl-H); 8.41 (s, 1H, phenyl-CHÄ
/
NH, D-exch.).
4.5.18. 2-(2-Nitrobenzyliden)hydrazo-4-methylthiazole
(51)
4.5.27. 2-(3,4-Dimethoxybenzyliden)hydrazo-4-
phenylthiazole (60)
1H NMR (DMSO): d 2.26 (s, 3H, CH3); 6.72 (s, 1H,
1H NMR (DMSO): d 3.90 (s, 6H, OCH3); 7.11ꢀ
/
7.97
(m, 8H, phenyl-H); 7.41 (s, 1H, H5-thiaz.); 8.08 (s, 1H,
phenyl-CHÄ); 11.70 (s, 1H, NH, D-exch.).
H5-thiaz); 7.50ꢀ
phenyl-CHÄ
/
8.47 (m, 4H, phenyl-H); 8.74 (s, 1H,
/); 11.75 (s, 1H, NH, D-exch.).
/
4.5.19. 2-(2-Nitrobenzyliden)hydrazo-4-phenylthiazole
(52)
4.5.28. 2-(4-Methylbenzyliden)hydrazo-4-methylthiazole
(61)
1H NMR (DMSO): d 7.42ꢀ
/
8.14 (m, 9H, phenyl-H);
7.51 (s, 1H, H5-thiaz.); 8.55 (s, 1H, phenyl-CHÄ
); 12.50
(s, 1H, NH, D-exch.).
1H NMR (DMSO): d 2.31 (s, 3H, CH3); 2.51 (s, 3H,
/
phenyl-CH3); 6.70 (s, 1H, H5-thiaz); 7.27ꢀ
/7.69 (m, 4H,
phenyl-H); 8.40 (s, 1H, phenyl-CHÄ
/); 10.83 (s, 1H, NH,
D-exch.).
4.5.20. 2-(3-Nitrobenzyliden)hydrazo-4-methylthiazole
(53)
4.5.29. 2-(4-Methylbenzyliden)hydrazo-4-phenylthiazole
(62)
1H NMR (DMSO): d 2.27 (s, 3H, CH3); 6.70 (s, 1H,
H5-thiaz.); 7.65ꢀ
phenyl-CHÄ
/
8.56 (m, 4H, phenyl-H); 8.64 (s, 1H,
); 11.63 (s, 1H, NH, D-exch.).
1H NMR (CDCl3): d 2.41 (s, 3H, CH3); 6.75 (s, 1H,
H5-thiaz.); 7.26 (s, 1H, phenyl-CHÄ
phenyl-H); 10.96 (s, 1H, NH, D-exch.).
/
); 7.43ꢀ7.74 (m, 9H,
/
/
4.5.21. 2-(3-Nitrobenzyliden)hydrazo-4-phenylthiazole
(54)
4.6. Microbiology
1H NMR (CDCl3); 7.50 (s, 1H, H5-thiaz.); 7.56ꢀ
/
8.34
); 10.36 (s,
(m, 9H, phenyl-H); 8.59 (s, 1H, phenyl-CHÄ
/
4.6.1. Compounds
Compounds for antimicrobial studies were dissolved
in dimethylsulfoxide at 10 mg/ml and stored at ꢁ20 8C.
1H, NH, D-exch.).
/
The working solutions were prepared in the same
medium used for the tests. To avoid interference with
the solvent [7], the highest DMSO concentration was
1%.
4.5.22. 2-(4-Nitrobenzyliden)hydrazo-4-methylthiazole
(55)
1H NMR (DMSO): d 2.25 (s, 3H, CH3); 6.70 (s, 1H,
H5-thiaz); 8.13ꢀ
CHÄ
/
8.36 (m, 5H, phenyl-H and phenyl-
/); 11.74 (s, 1H, NH, D-exch.).
4.6.2. Microorganisms
The antimicrobial activity of compounds 3ꢀ
/
27 and
4.5.23. 2-(4-Nitrobenzyliden)hydrazo-4-phenylthiazole
(56)
33ꢀ62 was evaluated against five Gram-positive species
/
(Staphylococcus aureus, S. epidermidis, Streptococcus
agalactiae, S. faecalis, and B. subtilis), and five Gram-
negative species (Escherichia coli, Pseudomonas aerugi-
nosa, Salmonella typhi, Proteus mirabilis, and Klebsiella
pneumoniae) isolated from clinical specimens.
1H NMR (CDCl3): d 6.84 (s, 1H, H5-thiaz.); 7.25ꢀ
/
8.30 (m, 9H, phenyl-H); 8.59 (s, 1H, phenyl-CHÄ
/
); 12.48
(s, 1H, NH, D-exch.).
C. albicans, C. glabrata, C. krusei, C. parapsilosis,
Saccharomyces cerevisiae, and C. tropicalis, were all
isolated from clinical specimens, and were used in the
evaluation of antifungal activity.
4.5.24. 2-(4-Methoxybenzyliden)hydrazo-4-
methylthiazole (57)
1H NMR (DMSO): d 2.26 (s, 3H, CH3); 3.80 (s, 3H,
OCH3); 6.71 (s, 1H, H5-thiaz.); 7.02ꢀ
phenyl-H); 8.43 (s, 1H, phenyl-CHÄ); 11.80 (s, 1H,
NH, D-exch.).
/7.75 (m, 4H,
/
4.6.3. Determination of MICs
The MICs of the compounds against Gram-positive
bacteria, Gram-negative bacteria, and fungi, were de-
termined by a standard broth macrodilution method
[8,9]. Tests with Gram-positive and Gram-negative
bacteria were carried out in Mueller Hunton broth
(Difco Laboratories, Detroit, MI). Antifungal activity
was evaluated in Sabouraud Dextrose broth (Difco
4.5.25. 2-(4-Methoxybenzyliden)hydrazo-4-
phenylthiazole (58)
1H NMR (DMSO): d 3.91 (s, 3H, OCH3); 7.42 (s, 1H,
H5-thiaz.) 7.10ꢀ
phenyl-CHÄ
/
7.97 (m, 9H, phenyl-H); 8.13 (s, 1H,
/); 11.42 (s, 1H, NH, D-exch.).

E. Maccioni et al. / Il Farmaco 58 (2003) 951ꢀ
/959
959
[4] A. Maccioni, E. Marongiu, Alcuni derivati del ciclopentanone*
Nota II, Ann. Chim. 49 (1959) 1283ꢀ1287.
[5] L. Birkofer, S.M. Kim, H.D. Engels, Aldehydaddition an
enamine, Chem. Ber. 95 (1962) 1495ꢀ1504.
[6] C. Anchisi, B.M. Baroli, A.M. Fadda, A.M. Maccioni, E.
Maccioni, C. Sinico, Synthesis of cis- and trans-2-arylmethylcy-
cloalkylamines: potential dopaminergic agents, Farmaco 52
/
Laboratories) [10]. In the case of fungi MFC (minimum
fungicidal concentration) values, or more commonly
MLC (minimum lethal concentration) values were also
measured.
/
/
The compounds were diluted in the test medium to
obtain a final concentration ranging between 100 and
0.19 mg/ml. Tubes containing 1 ml of the diluted
(1997) 55ꢀ59.
/
compounds were inoculated with 1ꢂ
105 bacteria and
/
[7] C. Jagannath, V.M. Reddy, P.R. Gangadharam, Enhancement of
drug susceptibility of multi-drug resistant strains of Mycobacter-
ium tuberculosis by ethambutol and dimethyl sulfoxide, J.
incubated at 37 8C for 18 or 24 h.
Antimicrob. Chemother. 35 (1995) 381ꢀ390.
/
[8] L.D. Trhupp, Susceptibility testing of antibiotics in liquid media,
in: V. Lorian (Ed.), Antibiotics in Laboratory Medicine, Williams
References
and Wilkins, Baltimore, MD, 1986, pp. 93ꢀ150.
/
[9] J.A. Washington, V.L. Sutter, Dilution susceptibility test: agar
and macro-broth dilution procedures, in: E.H. Lennette, E.H.
Spaulding, J.I. Truant (Eds.), Manual of Clinical Microbiology,
American Society for Microbiology, Washington, DC, 1980, pp.
[1] M.C. Cardia, M. Begala, A. DeLogu, E. Maccioni, A. Plumitallo,
Synthesis and antimicrobial activity of novel arylidenisothiosemi-
carbazones, Farmaco 55 (2000) 93ꢀ98.
/
[2] E. Maccioni, M.C. Cardia, L. Bonsignore, A. Plumitallo, M.L.
Pellerano, A. De Logu, Synthesis and anti-microbial activity of
isothiosemicarbazones and cyclic analogues, Farmaco 57 (2002)
533ꢀ544.
/
[10] M.A. Pfaller, M.G. Rinaldi, J.N. Galgiani, M.S. Bartlett, B.A.
Body, A. Espinel-Ingroff, R.A. Fromtling, G.S. Hall, C.E.
Hughes, F.C. Odds, A.M. Sugar, Collaborative investigation of
variables in susceptibility testing of yeasts, Antimicrob. Agents
809ꢀ817.
/
[3] G. Stork, A. Brizzolara, H. Landesman, J. Szmuszkovicz, R.
Terrel, The enamine alkylation and acylation of carbonyl
compounds, J. Am. Chem. Soc. 85 (1963) 207ꢀ
/222.
Chemother. 34 (1990) 1648ꢀ1654.
/