Synthesis, spectroscopic properties and antipathogenic activity of new thiourea derivatives

Article abstract of DOI:10.3390/molecules16097593

A number of acylthioureas, 2-((4-methylphenoxy)methyl)-N- (arylcarbamothioyl) benzamides (aryl = 3,5-dichlorophenyl, 2,3-dichlorophenyl, 3,4-dichlorophenyl, 2,4,5-trichlorophenyl, 3,4,5-trichlorophenyl, 2-bromophenyl, 2,4-dibromophenyl, 2,5-dibromophenyl,

Full text of DOI:10.3390/molecules16097593

Molecules 2011, 16, 7593-7607; doi:10.3390/molecules16097593
OPEN ACCESS
molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
Synthesis, Spectroscopic Properties and Antipathogenic Activity
of New Thiourea Derivatives
Carmen Limban 1, Luminita Marutescu 2 and Mariana Carmen Chifiriuc 2,3,
*
1
Pharmaceutical Chemistry Department, University of Medicine and Pharmacy Carol Davila,
Bucharest, Romania; E-Mail: carmen_limban@yahoo.com
2
3
Microbiology Department, Faculty of Biology, University of Bucharest, Ale. Portocalelor 1-4,
Bucharest 60101, Romania; E-Mail: lumi_dascalu@yahoo.com
National Institute for Research and Development in Microbiology and Immunology “Cantacuzino”,
Spl. Independentei 103, Bucharest 060031, Romania
* Author to whom correspondence should be addressed; E-Mail: carmen_balotescu@yahoo.com.
Received: 4 August 2011; in revised form: 29 August 2011 / Accepted: 1 September 2011 /
Published: 6 September 2011
Abstract:
A
number of acylthioureas, 2-((4-methylphenoxy)methyl)-N-(aryl-
carbamothioyl)benzamides (aryl = 3,5-dichlorophenyl, 2,3-dichlorophenyl, 3,4-dichloro-
phenyl, 2,4,5-trichlorophenyl, 3,4,5-trichlorophenyl, 2-bromophenyl, 2,4-dibromophenyl,
2,5-dibromophenyl, 2-iodophenyl, 3-fluorophenyl, 2,3,4-trifluorophenyl, 2,4,5-trifluoro-
phenyl, 2,4,6-trifluorophenyl) have been synthesized, characterized by elemental analysis,
IR and NMR spectroscopy and tested for their interaction with bacterial cells in free and
adherent state. The anti-pathogenic activity was correlated with the presence of one iodine,
bromide or fluorine, and two or three chloride atoms on the N-phenyl substituent of the
thiourea moiety, being significant especially on Pseudomonas aeruginosa and
Staphylococcus aureus strains, known for their ability to grow in biofilms. Our results
demonstrate the potential of these derivatives for further development of novel anti-
microbial agents with antibiofilm properties.
Keywords: acylthioureas; benzamides; 2-(4-methylphenoxymethyl)benzoic acid;
antimicrobial activity; bacterial biofilm

Molecules 2011, 16
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1. Introduction
Nowadays antibiotic resistance has become a serious public health problem. As a result of the rapid
development of resistance to the existing portfolio of antimicrobial drugs, there is an increasing need
to design new antibacterial and antifungal agents with better activity profiles and lower toxicity.
In recent years, thiourea derivatives have become a subject of interest due to their antibacterial,
antifungal, antimycobacterial and antileprotic activities. A thiourea system attached to a polycyclic
imide, such as thiourea derivatives of 4-azatricyclo[5.2.2.02,6]undec-8-ene-3,5-dione, showed
significant inhibitory activity against Gram-positive cocci [1]. Some pyridazine derivatives carrying
thiourea moieties exhibit potent inhibitory activity against Staphylococcus aureus, Escherichia coli,
Candida albicans and C. parapsilosis [2]. 1-Aroyl-3-arylthioureas showed moderate to potent activity
against different bacterial strains, especially against E. coli strains resistant to standard drugs [3]. The
antibacterial activity of the 1-aroyl-3-(substituted-2-benzothiazolyl)thioureas makes these substances
promising agents for the treatment of infections. [4]. N1,N2-disubstituted thioureas, such as N-phenyl-
and N-benzoylthioureas, have also attracted attention due to their potential application in medicinal
chemistry as antimicrobial agents [5].
Novel aroyl, thiophenoyl, morpholinoyl and butanoyl thiourea derivatives containing a thiazole
moiety, such as N-[(1,3-thiazol-2-ylamino)carbonothioyl]thiophene-2-carboxamide and N-[(1,3-
thiazol-2-ylamino)carbonothioyl]morpholine-4-carboxamide exhibit broad spectrum antifungal
activity [6]. For example, the 1-(isomeric fluorobenzoyl)-3-(isomeric fluorophenyl)thioureas were
proven to exhibit better antifungal than antibacterial activity, probably due to the inclusion of fluorine
that may increase the lipophilicity and enhance the rate of cell penetration and transport of the drug to
an active site. The higher polarizability due to the C-F bond may provide new possibilities for binding
to the receptor [7].
N-phenyl-N′-[4-(5-cyclohexylamino-1,3,4-thiadiazole-2-yl)phenyl]thiourea is active against
Mycobacterium tuberculosis H37Rv [8]. In an effort to improve the therapeutic value of thiocarlide,
several N-pentofuranosyl-N′′-[p-isoamyloxy)phenyl]thioureas were tested as potential anti-tuberculosis
therapeutic agents. N-D-Aldopentofuranosyl-N′-[p-(isoamyloxy)phenyl]thiourea derivatives, designed
as structural analogues of thiocarlide, have recently been reported to be more potent than thiocarlide
itself [9]. Novel 1-(5-cyclobutyl-1,3-oxazol-2-yl)-3(sub)phenyl/ pyridyl thiourea compounds inhibited
Mycobacterium tuberculosis H37Rv and a clinical isolate of multidrug- resistant M. tuberculosis [10].
Many articles have presented the progress in the investigation of new thiourea derivatives as
anthelmintics [11], antimalarials [12], antitumor agents [13], analgesics [14] and antivirals [15,16],
these compounds proving also useful as insecticides [17,18], fungicides [19], herbicides [20], and
plant-growth regulators [21]. Metals complexes with N-acyl thiourea ligands have been described in
the literature with regard to their synthesis, general characterization and biological activity, such as
cancerostatic activity [22].
Taking into account the improvement of the antimicrobial activity by the inclusion of fluorine in the
organic molecule of thioureas [7], we synthesized, characterized and studied the antimicrobial activity
of the new 2-((4-methylphenoxy)methyl)-N-(arylcarbamothioyl)benzamides, substituted with phenyl
groups carrying one, two or three atoms of F, Cl, Br and I.

Molecules 2011, 16
7595
2. Results and Discussion
2.1. Chemistry
Our decision to synthesize and evaluate this kind of substances for their antimicrobial activity was
influenced by several reports disclosing the antimicrobial activities of thiourea-based compounds. We
have previously synthesized and demonstrated the microbicidal activities of 2-(4-methoxyphenoxy-
methyl)benzoic acid [23], 2-(4-chlorophenoxymethyl)benzoic acid [24], 2-(4-fluorophenoxymethyl)-
benzoic acid [25], 2-(4-ethylphenoxymethyl)benzoic acid [26,27], and 2-(4-methylphenoxymethyl)-
benzoic acid thioureides [28].
Scheme 1. Synthesis of the new compounds.
-
CH O
+
HO
CH3
2
HCl
KCl
+
-
O
KOH
-
+
CH3
COO K
O
6
5
-
CH O
-
CH
O
+ SOCl2
2
+ NH4SCN
CH3
2
CH3
COOH
COCl
2
3
13
11
14
15
8
4
7
9
12
-
CH
O
3
CH2-O
H
CH3
+
R
H2N
2
5
6
10
2
1
N
S
R
CH3
C
C N=C=S
16 C
1a- m
O
O
4
N
H
Cl
Cl
Cl
Cl
Cl
19
Cl
18
,
,
,
,
,
Cl
Cl
20
Cl
R=
17
21
22
Cl
Cl
Cl
1e
1d
1b
1c
1a
Br
Br
I
Br
F
F
F
,
,
,
,
,
,
Br
F
Br
1j
1i
1g
1h
1k
1f
F
F
,
F
F
F
F
1m
1l

Molecules 2011, 16
7596
The new derivatives 1a-m were synthesized in good yields following the method described by us
(Scheme 1). 2-(4-Methylphenoxymethyl)benzoic acid (2) was converted into the corresponding acid
chloride 3 using thionyl chloride and anhydrous 1,2-dichloroethane as reaction medium, then treated
with a solution of ammonium thiocyanate in acetone to afford arylisothiocyanate 4 in situ, followed by
refluxing with amines to provide acylthioureas 1a-m in good yields. The compounds were purified by
recrystallization from isopropanol. The acid 2 was synthesized with the best yield by using phtalide 5,
which was treated with potassium para-cresolate in xylene under reflux. The obtained potassium salt 6
of 2-(4-methylphenoxymethyl)benzoic acid, due to its good solubility in 10% aqueous potassium
hydroxide solution allowed its facile separation from xylene. The acid 2 was then precipitated using a
mineral acid solution. The necessary potassium para-cresolate was obtained from the corresponding
phenol and potassium hydroxide in xylene. The resulting water was removed by azeotropic distillation.
The new compounds have all been characterized by their melting point, elemental analysis, infrared
and NMR spectral studies. All spectroscopic and elemental analyses data confirm the proposed
structures of the new compounds.
2.2. Biological Activity
2.2.1. Antimicrobial Activity
The emergence of bacterial resistance and multiresistance to antibiotics has been declared by the
European Center for Disease Control (ECDC) as one of the major public health problems, besides HIV
infection, tuberculosis and influenza. The increasing occurrence of multiresistant pathogenic bacterial
strains has gradually rendered traditional antimicrobial treatment ineffective. Consequently, there is a
pressing need to develop new antibiotics in order to overcome the bacterial resistance [29].
The qualitative screening of the susceptibility spectra of various microbial strains to the newly
synthesized compounds showed that only five of the tested compounds (1c, 1d, 1f, 1i and 1j) exhibited
an antimicrobial effect quantified by the occurrence of a growth inhibition zone (Table 1). The
compounds 1c (substituted with two chloride atoms in positions 3 and 4) and 1j (containing one
fluorine atom in position 3) exhibited activity against S. aureus ATCC 25923 strain, while 1i
(containing one iodine atom in position 2) exhibited activity against S. aureus 1694 clinical strain. The
compound 1d (substituted with three chloride atoms in positions 2, 4 and 5) exhibited anti-
Pseudomonas activity, and 1f (containing one bromide atom in position 2) was active on P. aeruginosa
and K. pneumoniae strains. The compound 1j was the only one active both on Gram negative
P. aeruginosa and the Gram positive S. aureus.
The results of the quantitative assay revealed that the serial dilutions of the compounds that proved
to be active on different microbial strains in the qualitative screening assay, exhibited a low inhibitory
activity on the bacterial growth, with high MIC values of 1,000–500 μg/mL (Table 2). Despite the high
MIC values obtained for the tested compounds, we further tested them for their anti-pathogenic
features, taking into account that, in many cases, the sub-inhibitory concentrations of the antimicrobial
substances interfere with the expression of different virulence features, such are adhesins or toxins [30].

Molecules 2011, 16
Table 1. Results of the qualitative screening of the susceptibility spectra of various
7597
microbial strains to the newly synthesized compounds (diameter of the growth inhibition
zone expressed in mm).
Microbial strain
Compound
1a
1b
1c
1d
1e
1f
1g
1h
1i
0
0
10
0
0
0
0
0
0
10
0
0
0
0
0
0
0
0
0
11
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
6
0
12
0
0
0
6
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
10
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
1j
1k
0
0
0
0
0
0
0
0
0
0
0
0
1l
1m
DMSO
0
0
0
Table 2. Results of the quantitative assay of MIC for the compounds proving to be active
in the qualitative screening on the respective bacterial strains (MIC values expressed in
μg/mL).
Compound
Strain
1c
1d
1f
1i
1j
S. aureus ATCC 25923
S. aureus 1694
500
500
500
K. pneumoniae 1771
P. aeruginosa 1671
500
500
1000
1000
2.2.2. Antipathogenic Activity
The genetic resistance of different microbial strains to numerous antimicrobial agents is amplified
when one organism is found growing in a biofilm. Antimicrobial resistance is a trait typical of most
biofilm organisms and it has been speculated that biofilms are the causative agent of up to 65% of
bacterial infections [31]. Biofilms are thought to become recalcitrant to antimicrobial assault through a
number of different mechanisms.

Molecules 2011, 16
7598
Giving the increasing risk of antimicrobial resistance, research is focusing on the discovery of
anti-pathogenic drugs that block the coordinated expression of virulence factors, such as adherence
capacity or production of toxins, thus rendering the bacteria harmless. Contrary to antibiotics, these
new anti-pathogenic drugs do not kill the bacteria, and since the survival of the bacteria is not
threatened by this approach, the development of resistance, like that to antibiotics, is not anticipated to
be a serious problem.
The inert substrata including the prosthetic medical devices represent risk factors for the occurrence
of biofilm associated infections. Taking into account the differences in physiology and susceptibility to
antibiotics of biofilm-embedded bacteria [32], the compounds active against different bacterial strains
in free state were further chosen and investigated concerning their efficiency against the adherent cells
grown in biofilms developed in plastic wells. The experimental model uses mini volumes and multiple
well plastic plates, allowing the simultaneous testing of a large spectrum of antibiotic concentrations.
Our results showed that the five compounds selected for the quantitative MIC assay proved to be
extremely active, inhibiting the ability of the S. aureus, Ps. aeruginosa and K. pneumoniae strains to
adhere and colonize the inert substratum at very low concentrations (with one exception, ranging
between 13 µg/mL and 27 µg/mL), probably due to their interference with adhesins expression
(Table 3). Thus, despite their high MICs, the tested substances seem to be promising as antipathogenic
compounds, which may be used in low concentrations for prophylaxis or therapy by themselves or in
combination with an antibiotic.
Table 3. Results of the quantitative assay of the influence of the tested compounds on the
adherence ability and biofilm development on the inert substratum (values of inhibitory
concentrations of biofilm development are expressed in μg/mL).
Compound
Strain
1c
1d
1f
1i
1j
S. aureus ATCC 25923
S. aureus 1694
13
437
13
K. pneumoniae 1771
P. aeruginosa 1671
27
13
13
13
3. Experimental
3.1. General
All chemicals used for the preparation of the compounds were of reagent grade quality and were
obtained from Merck, Fluka or Sigma- Aldrich. p-Methylphenol was used freshly distilled. Acetone
was dried over K2CO3 and then distilled and ammonium thiocyanate was treated by heating at 100 °C
before use. The necessary liquid amines were dried with potassium hydroxide and afterwards distilled.
Melting points were determined in open capillary tubes on an Electrothermal 9100 apparatus and
uncorrected. C, H, N and S analysis were carried out on a Perkin Elmer CHNS/O Analyzer Series II
2400 elemental analyzer. The room temperature attenuated total reflection Fourier transform infrared
(FT-IR ATR) spectra of the all synthesized compounds were registered using a Bruker Vertex 70
spectrophotometer. The IR bands are given as w – weak, m – medium, s – intense, vs – very intense.

Molecules 2011, 16
7599
13
1H-NMR and C-NMR spectra were recorded in DMSO-d6 on a Gemini 300BB instrument, at room
1
13
temperature, operating at 300 MHz for H and 75 MHz for C and a Unity Inova 400 instrument,
operating at 400 MHz for 1H and 100 MHz for 13C. Chemical shifts were given as δ values in parts per
million (ppm) relative to tetramethylsilane as internal standard. Coupling constants J were given in Hz.
Spin multiplets are given as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd (double
doublet), td (triple doublet) and br (broad). The chemical shifts for hydrogen and carbon atoms were
established also by G-COSY, G-HSQC, G-HMBC experiments. The 2-(4-methylphenoxymethyl)-
benzoic acid and 2-(4-methyl-phenoxymethyl)benzoyl chloride derivatives were obtained in good
yields similar to those reported in the previous article [33].
3.1.1. General Synthesis Procedure of the New Thioureides
A solution of 2-(4-methylphenoxymethyl)benzoyl chloride (0.01 mol) in acetone (15 mL) was
added to a solution of ammonium thiocyanate (0.01 mol) in acetone (5 mL). The reaction mixture was
heated under reflux for 1 h, and then cooled at the room temperature. A solution of primary amine
(0.01 mol) in acetone (2 mL) was added to the mixture and heated under reflux for 1 h. The thioureide
was precipitated after the cooled reaction mixture was poured into 500 mL water. The solid product
was purified by recrystallization from isopropanol with active carbon.
3.1.2. Spectral Data
2-((4-Methylphenoxy)methyl)-N-(3,5-dichlorophenylcarbamothioyl)benzamide (1a); yield 70%, mp
1
197.1–198.9 °C. H-NMR: 12.31 (s, 1H, NH, D2O exchangeable); 11.98 (br s, 1H, NH, D2O
exchangeable); 7.67 (d, J = 1.7 Hz, 2H, H-18, H-22); 7.61 (bd, J = 7.4 Hz, 1H, H-7); 7.58 (dd, J = 1.6 Hz,
J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.4 Hz, J = 7.4 Hz, 1H, H-5); 7.50 (t, J = 1.7 Hz, 1H, H-20); 7.46
(td, J = 1.4 Hz, J = 7.8 Hz, 1H, H-6); 7.05 (d, J = 8.6 Hz, 2H, H-11, H-13); 6.87 (d, J = 8.6 Hz, 2H,
13
H-10, H-14); 5.26 (s, 2H, H-8); 2.22 (s, 3H, H-15); C-NMR: 179.34 (C-16); 169.92 (C-1); 156.05
(C-9); 140.19 (C-3); 135.80 (C-19, C-21); 133.52 (C-2); 133.18 (C-12); 130.98 (C-5); 129.69 (C-11,
C-13); 129.58 (C-17); 128.35 (C-4); 128.23 (C-7); 127.68 (C-6); 125.51 (C-20); 123.07 (C-18, C-22);
114.42 (C-10, C-14); 67.41 (C-8); 19.99 (C-15); FT-IR (ν cm−1): 3278m; 3075w; 3018w; 2914w;
2857w; 1675m; 1621w; 1575m; 1543s; 1513vs; 1440m; 1386m; 1315m; 1292m; 1243s; 1160s;
1107w; 1042m; 988w; 885w; 854w; 815w; 798w; 752m; 689m; 651w; 607w; Anal. Calcd for
C22H18Cl2N2O2S (445.36): C, 59.33; H, 4.07; N, 6.29; S, 7.20%; Found: C, 59.65; H, 4.23; N, 6.34;
S 7.25%.
2-((4-Methylphenoxy)methyl)-N-(2,3-dichlorophenylcarbamothioyl)benzamide (1b); yield 65%, mp
1
152.4–153.2 °C; H-NMR: 12.43 (s, 1H, NH, D2O exchangeable); 12.12 (br s, 1H, NH, D2O
exchangeable); 7.87 (dd, J = 1.2 Hz, J = 8.2 Hz, 1H, H-22); 7.64 (dd, J = 1.2 Hz, J = 8.2 Hz, 1H, H-20);
7.61 (bd, J = 7.4 Hz, 1H, H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.4 Hz,
J = 7.4 Hz, 1H, H-5); 7.46 (td, J = 1.4 Hz, J = 7.8 Hz, 1H, H-6); 7.41 (t, J = 8.2 Hz, 1H, H-21); 7.05
(d, J = 8.6 Hz, 2H, H-11, H-13); 6.88 (d, J = 8.6 Hz, 2H, H-10, H-14); 5.27 (s, 2H, H-8); 2.21 (s, 3H,
13
H-15); C-NMR: 180.15 (C-16); 170.17 (C-1); 156.02 (C-9); 137.35 (C-3); 135.77 (C-2); 133.13
(C-12); 131.77 (C-19); 129.61 (C-17); 127.33 (C-18); 131.02 (C-5); 129.69 (C-11, C-13); 128.51

Molecules 2011, 16
7600
(C-20); 128.48 (C-4); 128.40 (C-7); 127.73 (C-21); 127.64 (C-6); 126.80 (C-22); 114.51 (C-10, C-14);
67.51 (C-8); 19.99 (C-15); FT-IR (ν cm−1): 3252w; 3120w; 3026w; 2912w; 2855w; 1680m; 1573m;
1510vs; 1444m; 1418m; 1381m; 1318m; 1294m; 1240s; 1197m; 1163s; 1124m; 1108m; 1038m;
968w; 873w; 856w; 818w; 804w; 777w; 747m; 727m; 696m; 658m; 607w; 571w; Anal. Calcd for
C22H18Cl2N2O2S (445.36): C, 59.33; H, 4.07; N, 6.29; S, 7.20%; Found: C, 59.68; H, 4.15; N, 6.24;
S 7.37%.
2-((4-Methylphenoxy)methyl)-N-(3,4-dichlorophenylcarbamothioyl)benzamide (1c); yield 55%, mp
1
172.1–174.3 °C; H-NMR: 12.33 (s, 1H, NH, D2O exchangeable); 11.98 (br s, 1H, NH, D2O
exchangeable); 7.97 (d, J = 2.3 Hz, 1H, H-18); 7.65 (d, J = 8.8 Hz, 1H, H-21); 7.54 (dd, J = 2.3 Hz,
J = 8.8 Hz, 1H, H-22); 7.61 (bd, J = 7.4 Hz, 1H, H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H, H-4);
7.57 (dd, J = 1.6 Hz, J = 8.2 Hz, 1H, H-22); 7.55 (td, J = 1.4 Hz, J = 7.4 Hz, 1H, H-5); 7.46 (td,
J = 1.4 Hz, J = 7.8 Hz, 1H, H-6); 7.05 (d, J = 8.6 Hz, 2H, H-11, H-13); 6.87 (d, J = 8.6 Hz, 2H,
13
H-10, H-14); 5.26 (s, 2H, H-8); 2.21 (s, 3H, H-15); C-NMR: 179.18 (C-16); 169.90 (C-1); 156.04
(C-9); 137.88 (C-18); 135.80 (C-2); 133.16 (C-12); 130.58 (C-19); 130.98 (C-5); 130.29 (C-22);
129.70 (C-11, C-13); 129.59 (C-17); 128.39 (C-4); 128.26 (C-7); 128.07 (C-20); 127.65 (C-6); 125.92
(C-18); 124.73 (C-21); 114.46 (C-10, C-14); 67.40 (C-8); 19.98 (C-15); FT-IR (ν cm-1): 3330m;
2969m; 1679m; 1601m; 1584m; 1505vs; 1469s; 1443s; 1384m; 1332m; 1293m; 1227s; 1173s; 1144s;
1050w; 1026s; 971w; 873w; 853w;821m; 787w; 749m; 709m; 664m; 608w; 567w; Anal. Calcd for
C22H18Cl2N2O2S (445.36): C, 59.33; H, 4.07; N, 6.29; S, 7.20%; Found: C, 59.76; H, 3.96; N, 6.21;
S 7.23%.
2-((4-Methylphenoxy)methyl)-N-(2,4,5-trichlorophenylcarbamothioyl)benzamide (1d); yield 58%, mp
1
171.8–173.4 °C; H-NMR: 12.47 (br s, 1H, NH); 12.20 (br s, 1H, NH); 8.28 (s, 1H, H-22); 7.97
(s, 1H, H-19); 7.62 (bd, J = 7.4 Hz, 1H, H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td,
J = 1.4 Hz, J = 7.4 Hz, 1H, H-5); 7.46 (td, J = 1.4 Hz, J = 7.5 Hz, 1H, H-6); 7.04 (d, J = 8.6 Hz, 2H,
13
H-11, H-13); 6.86 (d, J = 8.62 Hz, H, H-10, H-14); 5.26 (s, 2H, H-8); 2.21 (s, 3H, H-15); C-NMR:
179.89 (C-16); 170.23 (C-1); 156.01 (C-9); 135.75 (C-3); 135.49 (C-2); 133.11 (C-12); 131.05 (C-5);
130.40 (C-19); 129.66 (C-11, C-13); 129.58 (C-20); 129.32 (C-17); 129.31 (C-7); 128.71 (C-21);
128.48 (C-4); 128.27 (C-22); 127.87 (C-18); 127.77 (C-6); 114.44 (C-10, C-14); 67.51 (C-8); 19.99
(C-15); FT-IR (ν cm−1): 3553w; 3094w; 2989w; 2917w; 1678m; 1603w; 1566s; 1511vs; 1462m;
1446m; 1391w; 1364w; 1311m; 1260m; 1242s; 1165s; 1127w; 1078w; 1070w; 1052m; 1040m; 895w;
878w; 857w; 835w; 814w; 799m; 753m; 699m; 668w; 613w; 564w. Anal. Calcd for C22H17Cl3N2O2S
(479.80): C, 55.04; H, 3.57; N, 5.84; S, 6.68%; Found: C, 55.31; H, 3.59; N, 5.71; S 6.60%.
2-((4-Methylphenoxy)methyl)-N-(3,4,5-trichlorophenylcarbamothioyl)benzamide (1e); yield 67%, mp
158.7–160.8 °C. 1H-NMR: 12.16 (br s, 2H, NH); 7.90 (s, 2H, H-18, H-22); 7.62 (bd, J = 7.4 Hz, 1H,
H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.4 Hz, J = 7.4 Hz, 1H, H-5); 7.46 (td,
J = 1.4 Hz, J = 7.5 Hz, 1H, H-6); 7.05 (d, J = 8.6 Hz, 2H, H-11, H-13); 6.87 (d, J = 8.6 Hz, 2H, H-10,
13
H-14); 5.26 (s, 2H, H-8); 2.21 (s, 3H, H-15). C-NMR: 179.51 (C-16); 169.87 (C-1); 156.04 (C-9);
137.90 (C-3); 135.82 (C-2); 133.10 (C-12); 132.32 (C-19, C-21); 129.59 (C-17); 126.79 (C-20);
131.03 (C-5); 129.69 (C-11, C-13); 128.36 (C-7); 128.29 (C-4); 127.69 (C-6); 125.06 (C-18, C-22);
114.42 (C-10, C-14); 67.40 (C-8); 20.00 (C-15). FT-IR (ν cm−1): 3290w; 3079w; 2912w; 1682m;

Molecules 2011, 16
7601
1588w; 1561m; 1536m; 1509vs; 1431m; 1381m; 1309m; 1248s; 1238s; 1992w; 1169s; 1150m;
1053w; 1039m; 887w; 860w; 813w; 804w; 782w; 752m; 718w; 688w; 666m; 611w. Anal. Calcd for
C22H17Cl3N2O2S (479.80): C, 55.04; H, 3.57; N, 5.84; S, 6.68%; Found: C, 55.29; H, 3.52; N, 5.91;
S 6.76%.
2-((4-Methylphenoxy)methyl)-N-(2-bromophenylcarbamothioyl)benzamide (1f); yield 62%, mp
144.5–145.7 °C. 1H-NMR: 12.38 (br s, 1H, NH); 12.02 (br s, 1H, NH); 7.85 (dd, J = 1.5 Hz, J = 8.1 Hz,
1H, H-22); 7.71 (dd, J = 1.5 Hz, J = 8.1 Hz, 1H, H-19); 7.62 (dd, J = 1.7 Hz, J = 6.6 Hz, 1H,
H-7); 7.58 (dd, J = 2.4 Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.7 Hz, J = 8.9 Hz, 1H, H-5); 7.47
(ddd, J = 2.4 Hz, J = 6.6 Hz, J = 8.9 Hz, 1H, H-6); 7.43 (ddd, J = 1.5 Hz, J = 7.3 Hz, J = 8.1 Hz, 1H,
H-21); 7.24 (ddd, J = 1.5 Hz, J = 7.3 Hz, J = 8.1 Hz, 1H, H-20); 7.06 (d, J = 8.6 Hz, 2H, H-11, H-13);
6.88 (d, J = 8.6 Hz, 2H, H-10, H-14); 5.27 (s, 2H, H-8); 2.21 (br s, 3H, H-15). The doublet signal of
1
the protons H-11 and H-13 in the H-NMR spectrum is further split by the methyl group. Scalar
4
13
coupling is experimentally measured and the value has J(H11,H13-CH3)=0.6 Hz. C-NMR: 179.99
(C-16); 170.17 (C-1); 156.01 (C-9); 136.60 (C-3); 135.70 (C-2); 133.19 (C-12); 129.63 (C-17); 118.96
(C-18); 135.70 (C-19); 132.53 (C-20); 130.99 (C-5); 129.69 (C-11, C-13); 128.51 (C-7); 128.49 (C-
22); 128.42 (C-21); 127.73 (C-6); 127.64 (C-20); 114.52 (C-10, C-14); 67.45 (C-8); 19.99
(C-15). FT-IR (ν cm−1): 3232w; 3027w; 2918w; 2857w; 1675m; 1578w; 1513vs; 1441m; 1387w; 1317w;
1287w; 1242s; 1161s; 1042m; 853w; 801w; 742m; 707m; 672w; 608w. Anal. Calcd for C22H19BrN2O2S
(455.36): C, 58.02; H, 4.21; N, 6.15; S, 7.04%; Found: C, 57.89; H, 4.28; N, 6.11; S 7.16%.
2-((4-Methylphenoxy)methyl)-N-(2,4-dibromophenylcarbamothioyl)benzamide (1g); yield 67%, mp
1
169.3–170.6 °C. H-NMR: 12.33 (br s, 1H, NH); 12.10 (br s, 1H, NH); 7.95 (d, J = 1.4 Hz, 1H,
H-19); 7.80 (d, J = 8.6 Hz, 1H, H-22); 7.64 (dd, J = 1.4 Hz, J = 8.6 Hz, 1H, H-21); 7.62 (bd, J = 7.4
Hz, 1H, H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.4 Hz, J = 7.4 Hz, 1H, H-5);
7.46 (td, J = 1.4 Hz, J = 7.5 Hz, 1H, H-6); 7.05 (d, J = 8.6 Hz, 2H, H-11, H-13); 6.87 (d, J = 8.6 Hz,
13
2H, H-10, H-14); 5.26 (s, 2H, H-8); 2.21 (br s, 3H, H-15). C-NMR: 180.00 (C-16); 170.12 (C-1);
155.99 (C-9); 136.36 (C-3); 135.74 (C-2); 133.11 (C-12); 129.58 (C-17); 120.22 (C-20); 119.64 (C-18);
134.41 (C-19); 131.03 (C-5); 130.64 (C-21); 129.82 (C-22); 129.69 (C-4); 129.69 (C-11, C-13); 128.49
(C-7); 127.72 (C-6); 114.51 (C-10, C-14); 67.43 (C-8); 19.99 (C-15). FT-IR (ν cm−1): 3261w; 3130w;
3032w; 2913w; 2855w; 1679m; 1615w; 1560m; 1512vs; 1443w; 1376w; 1295w; 1246s; 1163s; 1078w;
1038m; 972w; 942w; 860w; 801w; 781w; 755w; 672m; 601w; 541w. Anal. Calcd for C22H18Br2N2O2S
(534.26): C, 49.46; H, 3.40; N, 5.24; S, 6.00%; Found: C, 49.79; H, 3.29; N, 5.33; S 5.89%.
2-((4-Methylphenoxy)methyl)-N-(2,5-dibromophenylcarbamothioyl)benzamide (1h); yield 69%, mp
160.2–161.5 °C. 1H-NMR: 12.39 (br s, 1H, NH); 12.12 (br s, 1H, NH); 8.04 (d, J = 2.1 Hz, 1H, H-22);
7.66 (d, J = 8.5 Hz, 1H, H-19); 7.62 (bd, J = 7.4 Hz, 1H, H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H,
H-4); 7.55 (td, J = 1.4 Hz, J = 7.4 Hz, 1H, H-5); 7.46 (td, J = 1.4 Hz, J = 7.5 Hz, 1H, H-6); 7.43 (dd,
J = 2.1 Hz, J = 8.5 Hz, 1H, H-20); 7.05 (d, J = 8.6 Hz, 2H, H-11, H-13); 6.87 (d, J = 8.6 Hz, 2H,
H-10, H-14); 5.26 (s, 2H, H-8); 2.20 (br s, 3H, H-15). 13C-NMR: 179.93 (C-16); 170.15 (C-1); 156.00
(C-9); 138.18 (C-3); 135.68 (C-2); 133.21 (C-12); 129.57 (C-17); 119.60 (C-18); 118.25 (C-21);
134.09 (C-19); 131.00 (C-5); 130.95 (C-20); 130.61 (C-22); 129.67 (C-11, C-13); 128.50 (C-4);
128.43 (C-7); 127.77 (C-6); 114.44 (C-10, C-14); 67.50 (C-8); 20.03 (C-15). FT-IR (ν cm−1): 3241m;

Molecules 2011, 16
7602
2997w; 2916w; 1675m; 1567m; 1513vs; 1395m; 1313m; 1240s; 1160s; 1075w; 1043m; 957w; 874w;
814w; 790m; 754m; 700m; 608w; 562w. Anal. Calcd for C22H18Br2N2O2S (534.26): C, 49.46; H, 3.40;
N, 5.24; S, 6.00%; Found: C, 49.21; H, 3.28; N, 5.15; S 6.07%.
2-((4-Methylphenoxy)methyl)-N-(2-iodophenylcarbamothioyl)benzamide (1i); yield 94%, mp
1
134.4–136.2 °C. H-NMR: 12.22 (br s, 1H, NH); 12.00 (br s, 1H, NH); 7.92 (dd, J = 1.4 Hz, J = 7.9
Hz, 1H, H-19); 7.65-7.55(m, 4H, H-4, H-5, H-7, H-22); 7.46 (td, J = 1.4 Hz, J = 7.5 Hz, 1H, H-6);
7.44 (td, J = 7.9 Hz, 1H, H-21); 7.07 (dd, J = 1.5 Hz, J = 7.9 Hz, 1H, H-20); 7.05 (d, J = 8.6 Hz, 2H,
13
H-11, H-13); 6.87 (d, J = 8.6 Hz, 2H, H-10, H-14,); 5.27 (s, 2H, H-8); 2.22 (s, 3H, H-15). C-NMR:
180.13 (C-16); 170.07 (C-1); 156.01 (C-9); 140.06 (C-17); 139.77 (C-19); 135.69 (C-3); 133.19 (C-2);
131.02 (C-5); 129.72 (C-11, C-13); 129.56 (C-12); 128.74 (C-22); 128.57 (C-20); 128.52 (C-4); 128.45
(C-7); 128.36 (C-21); 127.72 (C-6); 114.56 (C-10, C-14); 96.89 (C-18); 67.35 (C-8); 19.98 (C-15). FT-IR
(ν cm−1): 3389s; 3153w; 3029w; 2924w; 1684m; 1608w; 1511vs; 1485vs; 1379m; 1354m; 1308m; 1237s;
1142s; 1068w; 1019m; 877w; 795w; 743m; 716w; 653m; 595w; 515w. Anal. Calcd for C22H19IN2O2S
(502.36): C, 52.60; H, 3.81; N, 5.58; S, 6.38%; Found: C, 52.86; H, 3.97; N, 5.43; S 6.34%.
2-((4-Methylphenoxy)methyl)-N-(3-fluorophenylcarbamothioyl)benzamide (1j); yield 76%, mp
1
3
116.9–119 °C. H-NMR: 12.48 (br s, 1H, NH); 11.89 (br s, 1H, NH); 7.69 (bd, J(F-H)= 10.9 Hz, 1H,
H-18); 7.62 (bd, J = 7.4 Hz, 1H, H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.4
3
Hz, J = 7.4 Hz, 1H, H-5); 7.46 (td, J = 1.4 Hz, J = 7.5 Hz, 1H, H-6); 7.43 (dd, J(H-22-H-21)= 8.2 Hz,
3J(H-20-H-21)= 8.6 Hz, 1H, H-21); 7.34 (ddd, J = 1.1 Hz, J = 1.9 Hz, J = 8.2 Hz, 1H, H-22); 7.10 (tdd,
3J(F-H)= 8.6 Hz, 3J(H-20-H-21)= 8.6 Hz, 3J(H 20-H 21)= 1.9 Hz, J(H-20-H-18)= 1.4 Hz, 1H, H-20); 7.05 (d, J = 8.6
3
Hz, 2H, H-11, H-13); 6.87 (d, J = 8.6 Hz, 2H, H-10, H-14); 5.27 (s, 2H, H-8); 2.20 (s, 3H, H-15).
13C-NMR: 178.91 (C-16); 170.00 (C-1); 161.57 (d, J(F-C)= 241.0 Hz, C-19); 156.05 (C-9); 139.48 (C-3);
135.81 (C-2); 133.16 (C-12); 129.60 (C-17); 130.98 (C-5); 130.19 (C-21); 129.71 (C-11, C-13);
128.43 (C-7); 128.27 (C-4); 127.64 (C-6); 120.05 (C-22); 114.49 (C-10, C-14); 112.80 (d, J(F-C)= 21.2
Hz, C-20); 110.95 (d, J(F-C)= 25.7 Hz, C-18); 67.40 (C-8); 19.96 (C-15). FT-IR (ν cm−1): 3345w;
3129m; 2921w; 2857w; 1675m; 1601m; 1551s; 1504vs; 1445s; 1380w; 1331m; 1302m; 1283m;
1271m; 1256m; 1227vs; 1176m; 1140s; 1056w; 1024m; 985w; 958w; 899w; 860w; 829w; 800w;
781w; 727m; 669m; 605w; 555w; 512w. Anal. Calcd for C22H19FN2O2S (394.46): C, 66.99; H, 4.85;
N, 7.10; S, 8.13%; Found: C, 67.33; H, 4.72; N, 7.03; S 8.15%.
2-((4-Methylphenoxy)methyl)-N-(2,3,4-trifluorophenylcarbamothioyl)benzamide (1k); yield 71%, mp
1
111.2–112.3 °C. H-NMR: 12.13 (br s, 1H, NH); 12.10 (br s, 1H, NH); 7.63 (bd, J = 7.4 Hz, 1H,
H-7); 7.62- 7.54 (m, 3H, H-4, H-5, H-22); 7.47 (td, J = 1.4 Hz, J = 7.5 Hz, 1H, H-6); 7.36 (ddd,
J(F-20-H-21)= 9.5 Hz, 3J(H-22-H-21)= 8.2 Hz, 4J(H-22-H-21)= 2.3 Hz, 1H, H-21); 7.04 (d, J = 8.6 Hz, 2H, H-11,
H-13); 6.86 (d, J = 8.6 Hz, 2H, H-10, H-14); 5.27 (s, 2H, H-8); 2.22 (s, 3H, H-15). 13C-NMR: 179.89
(C-16); 170.23 (C-1); 156.01 (C-9); 135.74 (C-2); 135.49 (C-12); 135.75 (C-3); 133.11; 131.05 (C-5);
130.40 (C-19); 129.66 (C-11, C-13); 129.58; 129.32; 129.31 (C-7); 128.48 (C-4); 128.27 (C-22); 127.87;
127.77 (C-6); 114.44 (C-10, C-14); 67.51 (C-8); 19.99 (C-15). FT-IR (ν cm−1): 3243w; 2990w; 2927w;
1680m; 1620m; 1578m; 1511vs; 1477m; 1445m; 1385w; 1324s; 1237s; 1191s; 1178m; 1152m; 1054w;
1035m; 1005m; 902w; 857w; 804m; 783w; 758w; 741m; 673m; 604w. Anal. Calcd for C22H17F3N2O2S
(430.44): C, 61.39; H, 3.98; N, 6.51; S, 7.45%; Found: C, 61.57; H, 4.08; N, 6.59; S 7.48%.

Molecules 2011, 16
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2-((4-Methylphenoxy)methyl)-N-(2,4,5-trifluorophenylcarbamothioyl)benzamide (1l); yield 81%, mp
1
137.3–138.6 °C. H-NMR: 12.31 (br s, 1H, NH); 12.11 (br s, 1H, NH); 8.08 (m, 1H, H-22); 7.70 (td,
3J(F-18,20-H-19)= 10.6 Hz, 4J(F-21-H-19)= 7.4 Hz, 1H, H-19); 7.62 (bd, J = 7.4 Hz, 1H, H-7); 7.58 (dd, J = 1.6
Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.4 Hz, J = 7.4 Hz, 1H, H-5); 7.46 (td, J = 1.4 Hz, J = 7.5 Hz,
1H, H-6); 7.05 (d, J = 8.6 Hz, 2H, H-11, H-13); 6.87 (d, J = 8.6 Hz, 2H, H-10, H-14); 5.26 (s, 2H, H-
8); 2.21 (s, 3H, H-15). The resonance signal of H-22 is a multiplet, because it is coupled with all three
fluorine atoms with different constants, and the ratio between the diference of chemical shifts and
coupling constant value is less than 5 (Δν/J). 13C-NMR: 180.38 (C-16); 170.27 (C-1); 156.01
(C-9); 152.04 (ddd, J(F-18-C-18) = 243.8 Hz, J(F-20-C-18) = 7.4 Hz, J(F-21-C-18) = 2.2 Hz, C-18); 148.02 (ddd,
J(F-20-C-20) = 248.1 Hz, J(F-21-C-20) = 14.7 Hz, J(F-18-C-20) = 11.8 Hz, C-20); 145.73 (ddd, J(F-21-C-21) = 241.6 Hz,
2J(F-20-C-21)= 13.3 Hz, 3J(F-18-C-21) = 3.7 Hz, C-21); 135.84 (C-3); 133.02 (C-2); 131.08 (C-5); 129.71 (C-
11, C-13); 129.64 (C-12); 128.51 (C-7); 128.40 (C-4); 127.71 (C-6); 122.83 (m, C-17); 115.25 (dd,
2J(F-21-C-22) = 22.0 Hz, 3J(F-20-C-22) = 13.2 Hz, C-22); 114.55 (C-10, C-14); 105.93 (dd, 2J(F-18-C-19) = 21.9 Hz,
2J(F-20-C-19) = 26.3 Hz, C-19); 67.47 (C-8); 19.96 (C-15). FT-IR (ν cm−1): 3249m; 3075w; 2919m; 1676m;
1601w; 1563vs; 1508vs; 1438s; 1384w; 1316s; 1235s; 1215vs; 1204s; 1155s; 1102m; 1054m; 1036m;
942w; 868m; 829w; 804w; 786w; 748w; 703m; 677m; 635w; 585w. Anal. Calcd for C22H17F3N2O2S
(430.44): C, 61.39; H, 3.98; N, 6.51; S, 7.45%; Found: C, 61.71; H, 3.82; N, 6.58; S 7.47%.
2-((4-Methylphenoxy)methyl)-N-(2,4,6-trifluorophenylcarbamothioyl)benzamide (1m); yield 82%, mp
1
137.3–138.6 °C. H-NMR: 12.10 (br s, 1H, NH); 11.56 (br s, 1H, NH); 7.64 (bd, J = 7.4 Hz, 1H,
H-7); 7.58 (dd, J = 1.6 Hz, J = 7.4 Hz, 1H, H-4); 7.55 (td, J = 1.4 Hz, J = 7.4 Hz, 1H, H-5); 7.46 (td,
J = 1.4 Hz, J = 7.5 Hz, 1H, H-6); 7.30 (m, 2H, H-19, H-21, syst. AA’BB’C); 7.07 (d, J = 8.6 Hz, 2H,
13
H-11, H-13); 6.88 (d, J = 8.6 Hz, 2H, H-10, H-14); 5.26 (s, 2H, H-8); 2.22 (s, 3H, H-15). C-NMR:
3
182.69 (C-16); 170.26 (C-1); 160.08 (dt, J(F-C-20)= 248.2 Hz, J(F-18,22-C-20) = 15.0 Hz, C-20); 159.92
(ddd, J(F-18-C-18)= 257.8 Hz, J(F-20-C-18)= 6.6 Hz, J(F-22-C-18)= 12.3 Hz, C-18, C-22); 156.50 (C-9); 136.39
(C-3); 133.54 (C-2); 130.22 (C-12); 131.62 (C-5); 130.28 (C-11, C-13); 129.20 (C-7); 129.04 (C-4);
128.29 (C-6); 115.24 (C-10, C-14); 101.36 (t, J(F-C)= 27.8 Hz, C-19, C-21); 68.11 (C-8); 20.60 (C-15).
The C-19 and C-21 signal is a triplet due to their scalar coupling with the orto fluorine atoms. FT-IR
(ν cm−1): 3193m; 3071w; 2922w; 1673s; 1644w; 1603w; 1586w; 1527vs; 1509vs; 1447s; 1382w;
1341m; 1289w; 1243s; 1229s; 1174vs; 1122s; 1037s;998w; 853m; 810m; 787w; 753m; 733m; 672m;
509w. Anal. Calcd for C22H17F3N2O2S (430.44): C, 61.39; H, 3.98; N, 6.51; S, 7.45%; Found: C,
61.74; H, 3.79; N, 6.48; S 7.37%.
3.2. Biological Assays
3.2.1. Assessment of the Antimicrobial and Anti-Pathogenic Activity of the Newly Synthesized
Compounds
The in vitro qualitative screening of the antimicrobial activity was carried out by an adapted agar-
disk diffusion technique using a bacterial suspension of 0.5 McFarland obtained from 24 hour cultures
[33,34]. The antimicrobial activities of the newly synthesized compounds were determined against
ATCC reference microbial strains, i.e., S. aureus ATCC 25923, Enterococcus faecalis ATCC 29212,
E. coli ATCC 25922, as well as clinical strains, recently isolated from different clinical specimens, i.e.,

Molecules 2011, 16
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S. aureus 1694, Salmonella sp. 9246, K. pneumoniae 1771, P. aeruginosa 1671, C. albicans 128 and
Aspergillus niger IC 13534. The microbial strains identification was confirmed by aid of VITEK I
automatic system. VITEK cards for identification and susceptibility testing (GNS-522) were
inoculated and incubated according to the manufacturer's recommendations. The results were
interpreted by using software version AMS R09.1. The compounds were solubilized in dimethyl
sulfoxide to a final concentration of 10 mg/mL. The obtained solutions were further sterilized by
filtration. A volume of 10 μL of each tested compounds solution was distributed directly on the solid
medium previously seeded with the microbial inocula. The inoculated plates were incubated for 24 hrs
at 37 °C. Antimicrobial activity was assessed by measuring the growth inhibition zones diameters
expressed in mm [36,37]. The quantitative assay of the minimal inhibitory concentration (MIC,
μg/mL) was based on liquid medium two-fold microdilutions and performed in 96 multi-well plates
[34,35]. In this purpose, serial binary dilutions of the tested compounds (ranging between 1,000 and
0.975 µg/mL) were performed in a 200 μL volume of nutrient broth/YPG and each well was seeded
with 20 μL microbial inocula of 0.5 McFarland density. The plates were incubated for 24 hours at 37
°C for bacterial strains, respectively for 48 hours at 28 °C for fungal strains and MICs were read as the
last concentration of compound which inhibited the microbial growth.
3.2.2. Study of the Influence of the Selected Compounds on the Ability of P. aeruginosa, S. aureus
and K. pneumoniae to Colonize the Inert Substratum and Form Biofilms
The anti-biofilm activity of the tested compounds was tested by the microtiter method. For this
purpose, the microbial strains have been grown in the presence of two-fold serial dilutions of the tested
compounds performed in liquid nutrient broth/YPG, distributed in 96-well plates and incubated for
24 hours at 37 °C for bacterial strains, respectively for 48 hours at 28 °C for fungal strains. At the end
of the incubation period, the plastic wells were emptied, washed three times with phosphate buffered
saline (PBS), fixed with cold methanol and stained with 1% violet crystal solution for 30 minutes. The
biofilm formed on plastic wells was resuspended in 30% acetic acid. The intensity of the colored
suspensions was assessed by measuring the absorbance at 490 nm. The last concentration of the tested
compound that inhibited the development of microbial biofilm on the plastic wells was considered the
minimum inhibitory concentration of the biofilm development and was also expressed in μg/mL
[36,37].
4. Conclusions
New 2-((4-methylphenoxy)methyl)-N-(arylcarbamothioyl)benzamides were prepared from 2-(4-
methylphenoxymethyl)benzoyl chloride via isothiocyanate formation, followed by treatment with
various substituted amines. Our results showed that some of the newly synthesized thiourea derivatives
carrying aryl groups substituted with one iodine, bromide, fluorine or with two or three chloride atoms
could be used for the further development of novel antimicrobial agents with anti-biofilm properties.

Molecules 2011, 16
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Acknowledgments
The present study was supported by the financial aid of the following research projects: PN2
41043/2007, PN2 42150/2008 and Capabilities/ Module III –Romanian- Turkey bilateral project no. 378/
2010).
References and Notes
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of Thiourea Derivatives of Azatricyclo[5.2.2.02,6]undec-8-ene-3,5-dione. Arch. Pharm. Res. 2010,
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2. Doğruer, D.; Urlu, Ş.; Önkol, T.; Özçelik, B.; Şahin, M.F. Synthesis of some pyridazine
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Turk. J. Chem. 2010, 34, 57-65.
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antibacterial activity of some 1-aroyl-3-aryl-thioureas. Chemistry 2009, 18, 152-158.
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new 1-aroyl-3-(substituted-2-benzothiazolyl)thioureas. Pharmaceut. Chem. J. 2008, 42, 191-195.
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L.C.; Vencato, I.; Lariucci, C.; Sa´, F.P. Antimicrobial activity and structural study of
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Sample Availability: Samples of the compounds are available from the authors.
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