Synthesis and structure-antibacterial activity relationship studies of 4-substituted phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-thiones

Article abstract of DOI:10.1007/s00044-010-9457-4

The synthesis and characterization of a series of 4-substituted phenyl-4,5-dihydrobenzo[f][1,4]oxazepin- 3(2H)-thiones were presented. Preliminary in vitro antimicrobial activity of the compounds was assessed against a panel of microorganisms including S.

Full text of DOI:10.1007/s00044-010-9457-4

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2011) 20:1170–1180
DOI 10.1007/s00044-010-9457-4
ORIGINAL RESEARCH
Synthesis and structure–antibacterial activity relationship studies
of 4-substituted phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thiones
•
Hikmet Agirbas Berat Kemal Fatma Budak
•
Received: 9 May 2010 / Accepted: 25 September 2010 / Published online: 10 October 2010
Ó Springer Science+Business Media, LLC 2010
Abstract The synthesis and characterization of a series
of 4-substituted phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-
3(2H)-thiones were presented. Preliminary in vitro anti-
microbial activity of the compounds was assessed against a
panel of microorganisms including S. aureus, E. faecalis,
P. aeruginosa, E. coli, and C. albicans. Some of the
compounds exhibited significantly in vitro antimicrobial
activity. The pMIC values were correlated with physico-
chemical descriptors: Hammett substituent constants (rm
and rp) and the lipophilic constant (p). One statistical
significant 2D-QSAR model was obtained with para-
substituted compounds. The pMIC values were also cor-
related with some theoretical descriptors as independent
variables and four statistical significant 2D-QSAR models
were also obtained with meta-substituted compounds.
effective antimicrobial drugs is a very important objective
and many research programs have been directed toward the
design of new agents.
Oxazepine derivatives have been attracting much interest
due to the wide range of biological activities. Among these
activities, it is worth mentioning antithrombotic (Mishra
et al., 2010), antiepileptic (Pekcec et al., 2009), anticon-
vulsant (Sharma et al., 2008), antiinflammatory (Schridhar
et al., 1979; Verma et al., 2008), progesterone agonist (Dols
et al., 2008), antifungal (Serrano-Wu et al., 2002), antag-
onist and analgesic (Okada et al., 1994; Hallinan et al.,
1994), antipsychotic (Liegeois et al., 1994), anxiolytics
(Effland et al., 1982), antihistaminic (Sleevi et al., 1991),
antiaggregating (Aono et al., 1991), and epidermal growth
factor receptor(EGFR) tyrosine kinase inhibitory (Smith
et al., 2006) activities. Considering the structural charac-
teristics of the benzoxazepine-3-ones, the existence of
seven-membered heterocyclic ring system, fused aromatic
group and the group –N–C(=O)–, similar to protein amide
bond, it is reasonable to expect inherent physiological
activities. Therefore, the study of substituent effects on
antimicrobial activity of these compounds can give better
understanding of their structure–activity relationships.
In this research, we synthesized 4-substituted phenyl-4,5-
dihydrobenzo[f][1,4] oxazepin-3(2H)-thiones (Scheme 1)
to determine their antimicrobial activity against some bac-
teria and fungi and to observe the substituent effects on the
activity. We also applied 2D-QSAR analysis to see the
relations of the molecular descriptors with the activity.
Keywords Benzoxazepine derivatives Á
Antimicrobial activity Á QSAR
Introduction
The rapid development of drug resistance, the unsatisfac-
tory status of present treatments of bacterial and fungal
infections and the drug side effects limit the usage of most
antimicrobial agents. Hence, the synthesis of new and
H. Agirbas (&) Á B. Kemal
Faculty of Arts and Sciences, Department of Chemistry,
Kocaeli University, Umuttepe, 41300 Izmit, Turkey
e-mail: agirbas@kocaeli.edu.tr
Synthesis
F. Budak
Faculty of Medicine, Department of Microbiology,
Kocaeli University, Umuttepe, 41300 Izmit, Turkey
The synthesis of compounds (2–6) was carried out as illus-
trated in Scheme 1. 2-[(E)-(substituted phenylimino)
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Med Chem Res (2011) 20:1170–1180
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Scheme 1 Synthesis of 4-
substituted phenyl-4,5-
dihydrobenzo[f][1,4]oxazepin-
3(2H)-thiones
C6H4X
O
N
NH2C6H4X
NaBH4
OH
OH
(1)
(2)
C6H4X
C6H4X
NH
N
NaOH
ClCH2COCl
O
or K2CO3
OH
OH
(3)
Cl
(4)
X
---------------
a: p-N(CH3)2
b: p-OCH3
c: p-CH3
d: m-CH3
e: H
f: p-F
g: m-OCH3
h: p-I
C6H4X
C6H4X
N
N
P2S5
O
S
i: p-Cl
j: p-Br
O
O
(5)
(6)
k: m-Cl
l: m-CF3
m: m-NO2
n: p-NO2
methyl]phenols (2a–n) were obtained from the reaction of
salicylaldehyde (1) with substituted anilines. Then the imi-
nes (2) were reduced by NaBH4 to give 2-((substituted
phenylamino)methyl)phenols (3a–n) which were reacted
with chloroacetyl chloride to have the corresponding amides
(4a–n). 4-Substituted phenyl-4,5-dihydrobenzo[f][1,4]
oxazepin-3(2H)-ones (5a–n) were obtained in quantita-
tive yields under the basic treatment of the amides.
Compounds (5a–n) were treated with P2S5 to give corre-
sponding 4-substituted phenyl-4,5-dihydrobenzo[f][1,4]ox-
azepin-3(2H)-thiones (6a–n). All the compounds obtained
in this study were analyzed by their IR (Agirbas et al.,
Staphylococcus aureus, Enterococcus faecalis, Pseudo-
monas aeruginosa, Escherichia coli, and Candida albicans
by determining their minimal inhibitory concentrations
(MIC) by broth microdilution susceptibility tests (CLSI,
2002, 2006). The biological activity results of the com-
pounds are given in Table 1. Compound (6j) was found to
be the most active derivative against E. faecalis at MIC
value of 12.5 lg/ml among the tested compounds. All the
compounds showed antimicrobial activity with MIC values
between 25 and 100 lg/ml against E. faecalis and
P. aeruginosa. Most active compounds are 6f, 6g, 6h, 6j,
6l, and 6m against E. coli with the activity of 100 lg/ml.
Compounds 6g and 6j exhibited highest activity (MIC:
100 lg/ml) against C. albicans. All benzoxazepines (6a–
n) displayed low activity against S. aureus.
1
2009) and H NMR spectra and representative compounds
(6a–n) by elemental analysis.
Biological Activity
QSAR analysis
Fourteen of the new synthesized compounds (6a–
n) were evaluated for their in vitro antimicrobial activ-
We have performed linear regression studies for molecular
descriptors with the antimicrobial activity of compounds
ity against
a
panel of microorganisms including
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Med Chem Res (2011) 20:1170–1180
Table 1 Antimicrobial activities of compounds(6a–n)
Compound (substituent)
Antibacterial and antifungal activities, lg/ml (lmol/ml)
S. aureus
ATCC25983
E. faecalis
ATCC 29212
P. aeruginosa
ATCC 27853
E. coli
ATCC 25922
C. albicans
ATCC 90028
Ampicillin
Ciprofloxacin
6a (p-NMe2)
6b (p-OMe)
6c (p-Me)
6d (m-Me)
6e (H)
0.78
0.78
–
6.25
–
0.25
0.25
0.25
0.04
–
400 (1.34)
400 (1.40)
200 (0.74)
800 (2.97)
800 (3.13)
800 (2.93)
200 (0.70)
400 (1.05)
400 (1.38)
100 (0.30)
400 (1.38)
400 (1.24)
200 (0.67)
[1600 (5.33)
–
25 (0.08)
50 (0.18)
25 (0.09)
50 (0.19)
100 (0.39)
50 (0.18)
25 (0.09)
50 (0.13)
25 (0.09)
12.5 (0.04)
100 (0.35)
25 (0.08)
25 (0.08)
50 (0.17)
–
100 (0.34)
100 (0.35)
25 (0.09)
100 (0.37)
100 (0.39)
100 (0.37)
50 (0.18)
50 (0.13)
100 (0.35)
25 (0.08)
200(0.69)
50 (0.15)
25 (0.08)
50 (0.17)
–
200 (0.67)
200 (0.70)
200 (0.74)
400 (1.49)
200 (0.78)
100 (0.37)
100 (0.35)
100 (0.26)
400 (1.38)
100 (0.30)
200 (0.69)
100 (0.31)
100 (0.33)
200 (0.67)
–
800 (2.68)
800 (2.80)
200 (0.74)
800 (2.98)
800 (3.13)
400 (1.46)
100 (0.35)
800 (2.1)
800 (2.76)
100 (0.30)
800 (2.76)
400 (1.24)
200 (0.67)
800 (2.66)
0.25
6f (p-F)
6g (m-OMe)
6h (p-I)
6i (p-Cl)
6j (p-Br)
6k (m-Cl)
6l (m-CF3)
6m (m-NO2)
6n (p-NO2)
Fluconazole
(6a–n). Biological activity data, reported as MIC values
(Table 1), are transformed to pMIC(-logMIC) on a molar
basis used as dependent variables to obtain the linear
relationship. The pMIC values were first correlated with
physicochemical descriptors: Hammett substituent con-
stants (rm and rp) (Hansch et al., 1991) and the lipophilic
constant (p) (Hansch et al., 1973). Non-statistical signifi-
cant correlations were obtained when the descriptors were
studied as independent variables. However, only one sta-
tistical significant 2D-QSAR model was obtained (Table 2,
Eq. 1) with para-substituted compounds.
refractivity(MR), polarizability (polar), magnitude of
dipolar moment (l), and the calculated log of octanol–
water partition coefficient (clogP) of the compounds were
also computed by Hyperchem software (Table 3).
Energies of highest occupied molecular orbital (EHOMO
)
and lowest unoccupied molecular orbital (ELUMO) were
calculated using Gaussian 03W program package (Frisch
et al., 2004) by means of DFT (B3LYP) with the
6-311G(d,p) basis set (Table 3). Again, non-statistical
significant correlations were obtained when these theoret-
ical descriptors were studied as independent variables, but
four statistical significant 2D-QSAR models were also
obtained (Table 2, Eq. 2–5). The overall quality of the
obtained 2D-QSAR models was indicated by the correla-
tion coefficients (r and r2), the standard deviation (s) of the
regression equation, F value (F-statistical analysis; Fischer
In order to include theoretical descriptors to the SAR
study, a geometry of all the compounds (6a–n) has been
completely optimized by ab initio (RHF/3-21G) method
incorporated in the Hyperchem package (HyperChem,
2002). Surface area (SA), molecular volume(MV), molar
Table 2 Significant 2D-QSAR models obtained for antimicrobial activity of 4-substituted phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thiones (6a–n)
Equations Regression equation
Statistic parameter
r2
n
r
r2 adj
s
p
F
1
2
3
4
5
a
pMICS.a. = 0.6025( 0.18)p - 0.24( 0.19)rp - 0.38( 0.10)
9
0.817 0.668 0.557 0.224 0.036 6.040
pMICS.a. = -6.36( 4.54)ELUMO ? 0.21( 0.12)Polar - 7.26( 3.62)
pMICS.a. = -4.85( 4.58)ELUMO ? 0.01( 0.01)SA - 4.24( 1.91)
pMICP.a = -8.65( 5.10)ELUMO ? 0.01( 0.01)SA - 3.38( 2.12)
6a 0.819 0.671 0.452 0.215 0.189 3.061
6a 0.829 0.687 0.479 0.209 0.175 3.299
6a 0.846 0.716 0.527 0.233 0.151 3.790
pMICS.a = -22.94( 15.76)EHOMO - 0.33( 0.16)clogP - 4.85( 3.36) 6a 0.822 0.677 0.462 0.212 0.183 3.147
Only meta-substituted compounds
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Med Chem Res (2011) 20:1170–1180
1173
Table 3 Theoretical descriptors of 4-substituted phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-thiones 6a–n used for the regression analyses
3
SA (A )
3
MV (A )
3
MR (A )
3
Polar (A )
˚
˚
˚
˚
Compounds
clogP
EHOMO (au)
ELUMO (au)
l
6a
6b
6c
6d
6e
6f
-0.15
-0.19
0.96
320.98
292.71
280.21
280.17
256.53
261.77
292.62
285.31
273.93
278.33
273.85
292.64
288.33
288.34
276.37
255.32
247.45
247.54
231.01
233.36
255.36
260.66
245.90
252.95
245.98
254.50
249.71
249.59
99.32
92.07
89.98
89.98
85.70
85.83
92.07
98.02
90.42
93.23
90.42
90.91
91.92
91.92
35.08
32.53
31.89
31.89
30.06
29.37
32.53
35.09
31.99
32.68
31.99
31.62
31.90
31.90
-0.1967
-0.2064
-0.2080
-0.2086
-0.2098
-0.2137
-0.2071
–
-0.0417
-0.0464
-0.0472
-0.0476
-0.0488
-0.0528
-0.0459
–
6.91
5.94
6.23
6.05
6.19
6.31
6.33
6.27
6.62
6.27
6.48
6.99
8.12
8.08
0.96
0.80
0.20
6g
6h
6i
-0.19
1.32
0.58
-0.2155
-0.2153
-0.2156
-0.2180
-0.2224
-0.2243
-0.0545
-0.0543
-0.0544
-0.0566
-0.1019
-0.1027
6j
0.85
6k
6l
0.58
1.37
6m
6n
-0.01
-0.01
test) and the number of data points (n). The predictability
of each model was assessed using the cross-validated
correlation coefficient (r2 adj). For the structure-reactivity
models, a value of r2 adj above 0.45 was considered.
In order to evaluate the predictive power of the models,
the data was split into the training, and test sets. The
regression equations of the training sets gave good coeffi-
cient of determination (r2) and internal cross-validation (r2
adj) values (Table 4). A fair predicted values of the test
sets were also obtained.
the meta-substituted aromatic rings of the compounds could
better fit into the pocket in the receptor which should contain
polar groups that interact effectively with the meta substit-
uents. This may possibly be the reason to get Eqs. 2–5
(Table 2).
The correlation matrix for the descriptors was performed
and no cross-relations between the descriptors used in each
equation were obtained. Thus, these parameters are
orthogonal and allowing its safe use in the multilinear
regression relationship (Myers, 1987 and Draper and
Smith, 1981). Squared correlation matrix of theoretical
descriptors used in the equations is given in Table 5. All
the statistical calculations were performed by means of the
SigmaPlot program package.
Equation 1 (Table 2) may suggest that the resonance
effect, which is shown by para substituents, reveals to have
more influence on the 2D correlation with the electronic and
lipophilic descriptors against S. aureus. On the other hand,
Table 4 The results of the application of training and test sets to 2D-QSAR models (Eqs. 1–5 in Table 2)
Equations Training set Regression equation of training set Test set (MIC) Predicted value Statistic parameter
r2
r2 adj
s
1
6a, 6c, 6e 6j, 6n pMICS.a. = 1.05p - 0.61rp - 0.42
6b (400)
6f (800)
6h (400)
6i (400)
6g (200)
6l (400)
6g (200)
6l (400)
6g (200)
6l (400)
747
512
1519
590
136
161
150
215
561
899
24
0.986
0.972 0.081
2
3
4
5
6d, 6e, 6k, 6m
6d, 6e, 6k, 6m
6d, 6e, 6k, 6m
6d, 6e, 6k, 6m
pMICS.a. = -9.82ELUMO ? 0.09Polar - 3.80
pMICS.a. = -9.68ELUMO ? 0.004SA - 1.90
pMICP.a = -14.09ELUMO ? 0.0003SA - 0.27
0.891
0.838
0.819
0.993
0.672 0.181
0.515 0.219
0.456 0.296
0.979 0.046
pMICS.a = -75.55EHOMO - 0.33clogP - 16.64 6g (200)
6l (400)
755
r2 coefficient of determination, r2 adj internal cross-validation, s the standard deviation
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Synthesis of 2-((phenylamino)methyl)phenol (3e)
(general procedure)
Table 5 Squared correlation matrix of the theoretical descriptors
used in the QSAR study
ELUMO
Surface area 0.08
clogP 0.14
Polarizability 0.02
EHOMO 0.70
r2
1
2-[(E)-(phenylimino)methyl]phenol (2e) (25 mmol, 5 g)
was dissolved in 100 ml of methanol and dioxan at 1:1
ratio. NaBH4 (25 mmol, 0.8 g) was added to the stirred
solution until the yellow color of the Schiff base disap-
peared (1 h). Cold water was added to the solution to give
precipitate. The precipitate was recrystallized from meth-
anol to give 2-((phenylamino)methyl)phenol (3e) (4.25 g,
80%). Mp: 112–113°C. IR(KBr), m (cm-1): 3265 (N–H).
1H NMR (CDCl3), d (ppm): 3.89 (s, 1H, NH); 4.36 (s, 2H,
CH2–NH); 6.75–6.81 (m, aromatic, 2H); 6.84–6.97 (m,
aromatic, 4H); 7.12–7.15 (m, aromatic, 2H); 7.19–7.24 (m,
aromatic, 1H); 8.55 (s, 1H, OH).
1
0.06
0.67
0.08
1
0.19
1
0.0008 0.005
1
ELUMO Surface area clogP Polarizability EHOMO
Conclusion
In conclusion, a series of 14 new 4-substituted phenyl-
4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-thiones (6) were
synthesized from corresponding ones (5). The obtained
compounds have a great interest because there have been
no reports on the antimicrobial studies of these thiones in
the literature. In this study, the in vitro antimicrobial
activity of compounds (6) exhibited good results against
P. aeruginosa, one of the species that show the most dra-
matic resistance problems related to nosocomial infections
and multiresistant strains (Kiska et al., 1999). Moreover,
quantitative structure–activity relationship studies allowed
to draw the following conclusion about the antimicrobial
activity of the synthesized thiones: (i) only para-substi-
tuted thiones (6) showed a fair 2D correlation with the
electronic (rp) and lipophilic (p) descriptors against
S. aureus; (ii) meta-substituted thiones (6) gave fair 2D
correlations with some theoretical descriptors against
S. aureus and P. aeruginosa. The QSAR study has
provided key information regarding the structure of
4-substituted phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thiones which we believe will help to design more potent
antimicrobial compounds.
Spectroscopic and analytical data of compounds (3)
2-((4-Dimethyaminophenylamino)methyl)phenol (3a) Yield:
1
62%; mp: 74–75°C; IR (KBr), m (cm-1): 3329 (N–H); H
NMR (CDCl3), d (ppm): 2.86 (s, 6H, N(CH3)2); 3.47 (s,
1H, NH); 4.35 (s, 2H, CH2–NH); 6.69–6.82 (m, aromatic,
2H); 6.81–6.90 (m, aromatic, 4H); 7.08–7.11 (m, aromatic,
1H); 7.17–7.29 (m, aromatic, 1H); 8.61 (s, 1H, OH).
2-((4-Methoxyphenylamino)methyl)phenol (3b) Yield: 60%;
mp: 128–129°C; IR (KBr), m (cm-1): 3254 (N–H); 1H
NMR (CDCl3), d (ppm): 3.76 (s, 3H, OCH3); 4.37 (s, 2H,
CH2–NH); 6.81-6.83 (m, aromatic, 4H); 6.85–6.90 (m,
aromatic, 2H); 7.10–7.13 (m, aromatic, 1H); 7.18–7.21 (m,
aromatic, 1H).
2-((4-Methylphenylamino)methyl)phenol (3c) Yield: 70%;
1
mp: 122–124°C; IR (KBr), m (cm-1): 3261.74 (N–H); H
NMR (CDCl3), d (ppm): 2.26 (s, 3H, CH3); 4.34 (s, 2H,
CH2–NH); 6.71-6.75 (m, aromatic, 2H); 6.82–6.88 (m,
aromatic, 2H); 7.02–7.04 (m, aromatic, 2H); 7.09–7.12 (m,
aromatic, 1H); 7.16–7.22 (m, aromatic, 1H).
Experimental
Melting points were determined on Electrothermal 9200
apparatus and are uncorrected. The FTIR spectra were
recorded using Shimadzu 8201 spectrometer with KBr
technique, in the region 4000–400 cm-1 that was cali-
2-((3-Methylphenylamino)methyl)phenol (3d) Yield: 64%;
1
mp: 113–115°C; IR (KBr), m (cm-1): 3267.52 (N–H); H
1
brated by polystyrene. H NMR spectra were recorded on
NMR (CDCl3), d (ppm): 2.30 (s, 3H, CH3); 3.87 (s, 1H,
NH); 4.39 (s, 2H, CH2–NH); 6.63–6.66 (m, aromatic, 2H);
6.72–6.75 (m, aromatic, 1H); 6.84–6.90 (m, aromatic, 2H);
7.10–7.15 (m, aromatic, 1H); 7.19–7.24; 8.48 (s, 1H, OH).
Bruker DPX-400 (400 MHz) High Performance Digital
FT-NMR Spectrometer using CDCl3 with Me4Si as an
internal standard. Silica Gel (Fluka or Merck) were used
for column chromatography.
Schiff base (2) (Scheme 1) were synthesized according
to the literature (Vogel, 1972). A band for the azomethine
2-((4-Fluorophenylamino)methyl)phenol (3f) Yield: 60%;
1
mp: 122–123°C; IR (KBr),m (cm-1): 3259.81 (N–H); H
group was observed in IR spectrum at about 1620 cm-1
.
NMR (CDCl3), d (ppm): 3.89 (s, 1H, NH); 4.36 (s, 2H,
CH2–NH); 6.75-6.81 (m, aromatic, 2H); 6.84–6.97 (m,
aromatic, 4H); 7.12–7.24 (m, aromatic, 2H); 8.55 (s, 1H,
OH).
For the synthesis of compounds (3–5), literature methods
(Derieg and Sternbach, 1966; Davion et al., 2004) were
applied with slight modifications.
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1175
2-((3-Methoxyphenylamino)methyl)phenol (3g) Yield: 68%;
mp: 68–69°C; IR (KBr), m (cm-1): 3269.45 (N–H); 1H
NMR (CDCl3), d (ppm): 3.74 (s, 3H, OCH3); 3.95 (s, 1H,
NH); 4.37 (s, 2H, CH2–NH); 6.37-6.47 (m, aromatic, 3H);
6.84–6.89 (m, aromatic, 2H); 7.11–7.24 (m, aromatic, 3H);
8.24 (s, 1H, OH).
(s, 1H, NH); 6.65–6.68 (m, aromatic, 2H); 6.83–6.86 (m,
aromatic, 1H); 6.90–6.96 (m, aromatic, 1H); 7.19–7.25 (m,
aromatic, 2H); 8.08–8.11 (m, aromatic, 2H).
Synthesis of 2-Chloro-N-(2-hydroxybenzyl)-N-
phenylacetamide (4e) (general procedure)
2-((4-Iodophenylamino)methyl)phenol (3h) Yield: 66%;
1
2-[(E)-(phenylamino)methyl]phenol (3e) (20 mmol, 4 g)
was dissolved in benzene (150 ml). Triethylamine
(20 mmol, 2.05 g) was added to the solution. Chloroacetyl
chloride (20 mmol, 2.26 g) in benzene (50 ml) was added
dropwise and the reaction mixture was stirred for 2 h at
room temperature. Then, precipitated triethylammonium
chloride salt was filtered off and the filtrate was evaporated
under vacuo. The residual oily matter was crystallized from
diethyl ether to give 2-chloro-N-(2-hydroxybenzyl)-
N-phenylacetamide (4e) (4.82 g, 87%) Mp: 104–105°C.
IR (KBr), m (cm-1): 3115(OH), 1635(C=O). 1H NMR
(CDCl3), d (ppm): 3.83 (s, 2H, CH2–N–Ar); 4.79 (s, 2H,
CH2–Cl); 6.59–6.61 (m, aromatic, 1H); 6.66–6.72 (m,
aromatic, 1H); 6.98–7.01 (m, aromatic, 1H); 7.07–7.10
(m, aromatic, 2H); 7.20–7.25 (m, aromatic, 1H); 7.43–7.47
(m, aromatic, 3H); 9.07 (s, 1H, OH).
mp: 130–132°C; IR (KBr), m (cm-1): 3255.95 (N–H); H
NMR (CDCl3), d (ppm): 4.00 (s, 1H, NH); 4.36 (s, 2H,
CH2–NH); 6.56–6.61 (m, aromatic, 2H); 6.87–6.92 (m,
aromatic, 2H); 7.15–7.25 (m, aromatic, 2H); 7.47–7.52 (m,
aromatic, 2H); 7.82 (s, 1H, OH).
2-((4-Chlorophenylamino)methyl)phenol (3i) Yield: 70%;
1
mp: 122–123°C; IR (KBr), m (cm-1): 3257.88 (N–H); H
NMR (CDCl3), d (ppm): 3.97 (s, 1H, NH); 4.36 (s, 2H,
CH2–NH); 6.71–6.76 (m, aromatic, 2H); 6.85–6.91 (m,
aromatic, 2H); 7.13–7.24 (m, aromatic, 4H); 8.00 (s, 1H,
OH).
2-((4-Bromophenylamino)methyl)phenol (3j) Yield: 67%;
1
mp: 125–127°C; IR (KBr), m (cm-1): 3257.88 (N–H); H
NMR (CDCl3), d (ppm): 3.98 (s, 1H, NH); 4.37 (s, 2H,
CH2–NH); 6.68–6.73 (m, aromatic, 2H); 6.87–6.92 (m,
aromatic, 2H); 7.15–7.25 (m, aromatic, 2H); 7.30–7.35 (m,
aromatic, 2H); 7.90 (s, 1H, OH).
Spectroscopic and analytical data of compounds (4)
2-Chloro-N-(2-hydroxybenzyl)-N-(4-dimethylaminophenyl)
acetamide (4a) Yield: 72%; mp: 106–107°C; IR (KBr), m
(cm-1): 3144.07 (OH), 1627.97 (C=O); 1H NMR (CDCl3),
d (ppm): 3.00 (s, 6H, N(CH3)2); 3.87 (s, 2H, CH2–N–Ar);
4.74 (s, 2H, CH2–Cl); 6.65–6.71 (m, aromatic, 4H);
6.86–6.89 (m, aromatic, 2H); 6.97–7.00 (m, aromatic, 1H);
7.19–7.25 (m, aromatic, 1H); 9.22 (s, 1H, OH).
2-((3-Chlorophenylamino)methyl)phenol (3k) Yield: 63%;
1
mp: 112–114°C; IR (KBr), m (cm-1): 3259.81 (N–H); H
NMR (CDCl3), d (ppm): 4.01 (s, 1H, NH); 4.36 (s, 2H,
CH2–NH); 6.65–6.69 (m, aromatic, 1H); 6.79–6.80 (m,
aromatic, 1H); 6.83–6.92 (m, aromatic, 3H); 7.10–7.25 (m,
aromatic, 3H); 7.62 (s, 1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(4-methoxyphenyl)acetam-
ide (4b) Yield: 79%; mp: 133–134°C; IR (KBr), m
(cm-1): 3151.79 (OH), 1629.90 (C=O); 1H NMR (CDCl3),
d (ppm): 3.84 (s, 2H, CH2–N–Ar); 3.85 (s, 3H, OCH3);
4.75 (s, 2H, CH2–Cl); 6.61–6.73 (m, aromatic, 2H);
6.91–7.00 (m, aromatic, 5H); 7.20–7.25 (m, aromatic, 1H);
9.07 (s, 1H, OH).
2-((3-Trifluorophenylamino)methyl)phenol (3l) Yield: 76%;
mp: 80–82°C; IR (KBr), m (cm-1): 3269.45 (N–H); 1H
NMR (CDCl3), d (ppm): 4.15 (s, 1H, NH); 4.41 (s, 2H,
CH2–NH); 6.72–6.85 (m, aromatic, 1H); 6.88–6.97 (m,
aromatic, 2H); 7.03–7.08 (m, aromatic, 1H); 7.10–7.13
(m, aromatic, 1H); 7.18–7.26 (m, aromatic, 2H); 7.30–7.35
(m, aromatic, 1H); 7.47 (s, 1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(4-methylphenyl)acetam-
ide (4c) Yield: 59%; mp: 102–104°C; IR (KBr),m (cm-1):
3169.15 (OH), 1629.90 (C=O); 1H NMR (CDCl3), d (ppm):
2.41 (s, 1H, CH3); 3.83 (s, 2H, CH2–N–Ar); 4.77 (s, 2H,
CH2–Cl); 6.61–6.70 (m, aromatic, 2H); 6.94–7.00 (m,
aromatic, 3H); 7.22–7.26 (m, aromatic, 3H); 9.10 (s, 1H,
OH).
2-((3-Nitrophenylamino)methyl)phenol (3m) Yield: 97%;
1
mp: 117–118°C; IR (KBr), m (cm-1): 3263.66 (N–H); H
NMR (CDCl3), d (ppm): 4.37 (s, 1H, NH); 4.44 (s, 2H,
CH2–NH); 6.69–6.82 (m, aromatic, 2H); 6.81–6.90
(m, aromatic, 4H); 7.08–7.11 (m, aromatic, 1H); 7.17–7.29
(m, aromatic, 1H); 8.61 (s, 1H, OH).
2-((4-Nitrophenylamino)methyl)phenol (3n) Yield: 64%;
1
2-Chloro-N-(2-hydroxybenzyl)-N-(3-methylphenyl)acetam-
ide (4d) Yield: 47%; mp: 102–104°C; IR (KBr), m
(cm-1): 3126.71 (OH), 1631.83 (C=O); 1H NMR (CDCl3),
mp: 135–136°C; IR (KBr), m (cm-1): 3365.90 (N–H); H
NMR (CDCl3), d (ppm); 4.46 (s, 2H, CH2–NH); 4.85
123

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Med Chem Res (2011) 20:1170–1180
d (ppm): 2.37 (s, 1H, CH3) 3.84 (s, 2H, CH2–N–Ar); 4.78
(s, 2H, CH2–Cl); 6.61–6.72 (m, aromatic, 2H); 6.83–6.92
(m, aromatic, 2H); 6.98–7.01 (m, aromatic, 1H); 7.20–7.35
(m, aromatic, 3H); 9.12 (s, 1H, OH).
6.95–7.00 (m, aromatic, 2H); 7.16–7.17 (m, aromatic, 1H);
7.21–7.27 (m, aromatic, 1H); 7.37–7.48 (m, aromatic, 2H);
8.86 (s, 1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(4-trifluorophenyl)acetam-
ide (4l) Yield: 47%; mp: 93–94°C; IR (KBr), m (cm-1):
3275.24 (OH), 1639.55 (C=O); 1H NMR (CDCl3), d (ppm):
3.80 (s, 2H, CH2–N–Ar); 4.81 (s, 2H, CH2–Cl); 6.56–6.59
(m, aromatic, 1H); 6.68–6.74 (m, aromatic, 1H); 6.98–7.01
(m, aromatic, 1H); 7.22–7.29 (m, aromatic, 2H); 7.41 (m,
aromatic 1H); 7.58–7.64 (m, aromatic, 1H); 7.73–7.76 (m,
aromatic, 1H); 8.78 (s, 1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(4-fluorophenyl)acetamide
(4f) Yield: 45%; mp: 91–93°C; IR (KBr), m (m (cm-1):
3221.23 (OH), 1633.76 (C=O); 1H NMR (CDCl3), d (ppm):
3.82 (s, 2H, CH2–N–Ar); 4.77 (s, 2H, CH2–Cl); 6.59–6.62
(m, aromatic, 1H); 6.68–6.73 (m, aromatic, 1H); 6.97–7.07
(m, aromatic, 1H); 7.08–7.26 (m, aromatic, 5H); 8.91 (s,
1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(3-methoxyphenyl)acetam-
ide (4g) Yield: 49%; mp: 69–70°C; IR (KBr), m (cm-1):
3157.58 (OH), 1633.76 (C=O); 1H NMR (CDCl3), d (ppm):
3.77 (s, 1H, OCH3) 3.87 (s, 2H, CH2–N–Ar); 4.78 (s, 2H,
CH2–Cl); 6.58–6.60 (m, aromatic, 1H); 6.64–6.74 (m,
aromatic, 3H); 6.97–7.00 (m, aromatic, 2H); 7.20–7.26 (m,
aromatic, 1H); 7.32–7.38 (m, aromatic, 1H); 9.06 (s, 1H,
OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(3-nitrophenyl)acetamide
(4m) Yield: 63%; mp: 105–106°C; IR (KBr), m (cm-1):
3221.23 (OH), 1635.69 (C=O); 1H NMR (CDCl3), d (ppm):
3.83 (s, 2H, CH2–N–Ar); 4.85 (s, 2H, CH2–Cl); 6.56–6.60
(m, aromatic, 1H); 6.68–6.74 (m, aromatic, 1H); 6.97–7.00
(m, aromatic, 1H); 7.22–7.28 (m, aromatic, 1H); 7.41–7.45
(m, aromatic, 1H); 7.65–7.70 (m, aromatic, 1H); 8.06–8.08
(m, aromatic, 1H); 8.33–8.36 (m, aromatic, 1H); 8.59 (s,
1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(4-iodophenyl)acetamide
(4h) Yield: 30%; mp: 122–123°C; IR (KBr), m (cm-1):
3178.79 (OH), 1643.41 (C=O); 1H NMR (CDCl3), d (ppm):
3.82 (s, 2H, CH2–N–Ar); 4.77 (s, 2H, CH2–Cl); 6.60–6.62
(m, aromatic, 1H); 6.63–6.74 (m, aromatic, 1H); 6.82–6.85
(m, aromatic, 2H); 6.97–7.00 (m, aromatic, 1H); 7.21–7.26
(m, aromatic, 1H); 7.78–7.81 (m, aromatic, 2H); 8.87 (s,
1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(4-nitrophenyl)acetamide
(4n) Yield: 40%; mp: 109–111°C; IR (KBr), m (cm-1):
3180.72 (OH), 1643.41 (C=O); 1H NMR (CDCl3), d (ppm):
3.84 (s, 2H, CH2–N–Ar); 4.85 (s, 2H, CH2–Cl); 6.56–6.59
(m, aromatic, 1H); 6.68–6.74 (m, aromatic, 1H); 6.97–7.00
(m, aromatic, 1H); 7.22–7.27 (m, aromatic, 1H); 7.32–7.36
(m, aromatic, 2H); 8.32–8.36 (m, aromatic, 2H); 8.59 (s,
1H, OH).
2-Chloro-N-(2-hydroxybenzyl)-N-(4-chlorophenyl)acetam-
ide (4i) Yield: 41%; mp: 115–117°C; IR (KBr), m (cm-1):
3221.23 (OH), 1637.62 (C=O); 1H NMR (CDCl3), d (ppm):
3.82 (s, 2H, CH2–N–Ar); 4.77 (s, 2H, CH2–Cl); 6.60–6.62
(m, aromatic, 1H); 6.69–6.71 (m, aromatic, 1H); 6.97–7.06
(m, aromatic, 3H); 7.21–7.27 (m, aromatic, 1H); 7.42–7.46
(m, aromatic, 2H); 8.88 (s, 1H, OH).
Synthesis of 4-phenyl-4,5-dihydrobenzo[f]
[1,4]oxazepin-3(2H)-one (5e) (general procedure,
except 5n)
2-Chloro-N-(2-hydroxybenzyl)-N-phenylacetamide
(4e)
(16 mmol, 4.4 g) was dissolved in 100 ml of ethanol and
13 ml of 5% NaOH solution was added. The reaction
mixture was stirred for 1 h at room temperature. To sep-
arate the inorganic materials, water was added to the
solution and the organic part was extracted with chloro-
form. Then, the organic layer was dried with anhydrous
CaCl2. The solvent was evaporated under vacuo. The
residual matter was crystallized from ethanol to give
4-phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-one (5e)
(2.1 g, 55%). Mp: 148–149°C, Lit. (Derieg and Sternbach,
2-Chloro-N-(2-hydroxybenzyl)-N-(4-bromophenyl)acetam-
ide (4j) Yield: 51%; mp: 113–115°C; IR (KBr), m (cm-1):
3201.94 (OH), 1643.41 (C=O); 1H NMR (CDCl3), d (ppm):
3.82 (s, 2H, CH2–N–Ar); 4.77 (s, 2H, CH2–Cl); 6.59–6.62
(m, aromatic, 1H); 6.68–6.73 (m, aromatic, 1H); 6.96–7.00
(m, aromatic, 3H); 7.21–7.26 (m, aromatic, 1H); 7.58–7.61
(m, aromatic, 2H); 8.86 (s, 1H, OH).
1
2-Chloro-N-(2-hydroxybenzyl)-N-(3-chlorophenyl)acetam-
ide (4k) Yield: 50%; mp: 114–116°C; IR (KBr), m
(cm-1): 3128.64 (OH), 1637.62 (C=O); 1H NMR (CDCl3),
d (ppm): 3.84 (s, 2H, CH2–N–Ar); 4.78 (s, 2H, CH2–Cl);
6.61–6.64 (m, aromatic, 1H); 6.69–6.75 (m, aromatic, 1H);
1966): 147–148°C; IR (KBr), m (cm-1): 1661(C=O); H
NMR (CDCl3), d (ppm): 4.87 (s, 2H, N–CH2); 4.89 (s, 2H,
O–CH2); 7.06–7.13 (m, aromatic, 2H); 7.16–7.25 (m,
aromatic, 1H); 7.26–7.34 (m, aromatic, 3H); 7.36–7.44 (m,
aromatic, 3H).
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Med Chem Res (2011) 20:1170–1180
1177
4-(4-Nitrophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)
one (5n) 2-Chloro-N-(2-hydroxybenzyl)-N-(4-nitrophenyl)
acetamide (4n) (2 mmol, 0.5 g) was dissolved in 100 ml of
acetone and K2CO3 (6 mmol, 0.83 g) was added. The reac-
tion mixture was refluxed for 2 h. K2CO3 was filtered off and
acetone was evaporated. The residue was washed with water
and then dissolved in chloroform. The solution was dried
with anhydrous CaCl2 and the solvent was evaporated under
vacuo. The residual matter was crystallized from ethanol
to give 4-(4-nitrophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-
3(2H)-one (5n) (0.25 g, 35 %). Mp: 194–195°C; IR (KBr), m
(cm-1): 1676.20 (C=O); 1H NMR (CDCl3), d (ppm): 4.91 (s,
2H, N–CH2); 4.98 (s, 2H, O–CH2); 7.09–7.14 (m, aromatic,
2H); 7.20–7.22 (m, aromatic, 1H); 7.34–7.40 (m, aromatic,
1H); 7.46–7.50 (m, aromatic, 2H); 8.26–8.29 (m, aromatic,
2H).
aromatic, 4H); 7.16–7.24 (m, aromatic, 3H); 7.32–7.37 (m,
aromatic, 1H).
4-(3-Methoxyphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5g) Yield: 63%; mp: 104–105°C; IR (KBr), m
1
(cm-1): 1666.55 (C=O); H NMR (CDCl3), d (ppm): 3.80
(s, 3H, OCH3) 4.86 (s, 2H, N–CH2); 4.88 (s, 2H, O–CH2);
6.79–6.86 (m, aromatic, 3H); 7.08–7.19 (m, aromatic, 3H);
7.29–7.37 (m, aromatic, 2H).
4-(4-Iodophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-one
(5h) Yield: 57%; mp: 176–178°C; IR (KBr), m (cm-1):
1
1668.48 (C=O); H NMR (CDCl3), d (ppm): 4.85 (s, 2H,
N–CH2); 4.86 (s, 2H, O–CH2); 7.06–7.18 (m, aromatic,
5H); 7.32–7.37 (m, aromatic, 1H); 7.51–7.55 (m, aromatic,
2H).
4-(4-Chlorophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5i) Yield: 45%; mp: 154–155°C; IR (KBr), m
Spectroscopic and analytical data of compounds (5)
1
(cm-1): 1668.48 (C=O); H NMR (CDCl3), d (ppm): 4.85
4-(4-Dimethylaminophenyl)-4,5-dihydrobenzo[f][1,4]oxaze-
pin-3(2H)-one (5a) Yield: 62%; mp: 213–214°C; IR
(KBr), m (cm-1): 1660.77 (C=O); 1H NMR (CDCl3), d
(ppm): 2.95 (s, 6H, N(CH3)2); 4.85 (s, 2H, N–CH2); 4.86
(s, 2H, O–CH2); 6.70–6.73 (m, aromatic, 2H); 7.03–7.17
(m, aromatic, 5H); 7.29-7.35 (m, aromatic, 1H).
(s, 2H, N–CH2); 4.86 (s, 2H, O–CH2); 7.08–7.12 (m, aro-
matic, 2H); 7.15–7.21 (m, aromatic, 3H); 7.32–7.39 (m,
aromatic, 3H).
4-(4-Bromophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5j) Yield: 60%; mp: 161–163°C; IR (KBr),m (cm-1):
1
1668.05 (C=O); H NMR (CDCl3), d (ppm): 4.85 (s, 2H,
4-(4-Methoxyphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5b) Yield: 80%; mp: 175–176°C, Lit. (Davion et al.,
2004, 175°C); IR (KBr), m (cm-1): 1653.05 (C=O); 1H
NMR (CDCl3), d (ppm): 3.81 (s, 3H, OCH3); 4.85 (s, 2H,
N–CH2); 4.85 (s, 2H, O–CH2); 6.90–6.93 (m, aromatic,
2H); 7.07–7.18 (m, aromatic, 5H); 7.30–7.34 (m, aromatic,
1H).
N–CH2); 4.86 (s, 2H, O–CH2); 6.99–7.03 (m, aromatic,
2H); 7.05–7.12 (m, aromatic, 2H); 7.15–7.18 (m, aromatic,
1H); 7.31–7.37 (m, aromatic, 1H); 7.70–7.75 (m, aromatic,
2H).
4-(3-Chlorophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5k) Yield: 71%; mp: 129–130°C; IR (KBr), m
1
(cm-1): 1668.48 (C=O); H NMR (CDCl3), d (ppm): 4.85
(s, 2H, N–CH2); 4.87 (s, 2H, O–CH2); 7.07–7.20 (m, aro-
4-(4-Methylphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5c) Yield: 50%; mp: 142–143°C; IR (KBr), m
1
matic, 4H); 7.26–7.38 (m, aromatic, 4H).
(cm-1): 1654.98 (C=O); H NMR (CDCl3), d (ppm): 2.36
(s, 3H, CH3); 4.85 (s, 2H, N–CH2); 4.86 (s, 2H, O–CH2);
7.05–7.16 (m, aromatic, 5H); 7.17–7.22 (m, aromatic 2H);
7.30–7.36 (m, aromatic 1H).
4-(3-Trifluoromethylphenyl)-4,5-dihydrobenzo[f][1,4]oxaze-
pin-3(2H)-one (5l). Yield: 49%; mp: 95–96°C; IR (KBr),
m (cm-1): 1670.41 (C=O); 1H NMR (CDCl3), d (ppm): 4.87
(s, 2H, N–CH2); 4.91 (s, 2H, O–CH2); 7.10–7.25 (m, aro-
matic, 3H); 7.34–7.39 (m, aromatic, 1H); 7.44–7.47 (m,
aromatic, 1H); 7.51–7.55 (m, aromatic, 3H).
4-(3-Methylphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5d) Yield: 80%; mp: 123–125°C; IR (KBr), m
1
(cm-1): 1668.48 (C=O); H NMR (CDCl3), d (ppm): 2.36
(s, 3H, CH3) 4.85 (s, 2H, N–CH2); 4.86 (s, 2H, O–CH2);
4-(3-Nitrophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5m) Yield: 60%; mp: 117–118°C; IR (KBr), m
7.02–7.18 (m, aromatic, 5H); 7.26–7.36 (m, aromatic, 3H).
1
(cm-1): 1680.05 (C=O); H NMR (CDCl3), d (ppm): 4.89
4-(4-Fluorophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
one (5f) Yield: 65 %; mp: 139–140°C; IR (KBr),m
(s, 2H, N–CH2); 4.96 (s, 2H, O–CH2); 7.09–7.15 (m, aro-
matic, 2H); 7.20–7.23 (m, aromatic, 1H); 7.34–7.40 (m,
aromatic, 1H); 7.55–7.66 (m, aromatic, 2H); 8.13–8.17 (m,
aromatic, 2H).
1
(cm-1): 1662.69 (C=O); H NMR (CDCl3), d (ppm): 4.85
(s, 2H, N–CH2); 4.86 (s, 2H, O–CH2); 7.06–7.13 (m,
123

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Synthesis of 4-phenyl-4,5-dihydrobenzo[f]
[1,4]oxazepin-3(2H)-thione (6e) (general procedure)
Anal. Calcd for C16H15NOS: C, 71.34; H, 5.61; N, 5.20;
Found: C, 71.46; H, 5.67; N, 5.02.
4-Phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-one (5e)
(6 mmol, 1.45 g) was dissolved in 150 ml of xylene. P2S5
(3 mmol, 0.67 g) was added to the solution. The reaction
mixture was refluxed for 3 h at 140°C and then filtered
instantly. The filtrate was evaporated under vacuo. The
residual oily matter was subjected to flash column chroma-
tography (eluent, ethyl acetate:petroleum ether, 1:6) and
crystallized from ethyl acetate:petroleum ether (1:9) to
give 4-phenyl-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-thi-
one (6e) (0.95 g, 62%). Mp: 102–103°C. Selected IR data
4-(4-Fluorophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6f) Yield: 21%; mp: 127–128°C; IR (KBr), m
1
(cm-1): 1346.36 (C=S); H NMR (CDCl3), d (ppm): 5.10
(s, 2H, N–CH2); 5.34 (s, 2H, O–CH2); 7.01–7.22 (m, aro-
matic, 7H); 7.29–7.33 (m, aromatic, 1H) 7.30–7.40 (m,
aromatic, 1H). Anal. Calcd for C15H12FNOS: C, 65.91; H,
4.43; N, 5.12; Found: C, 65.75; H, 4.67; N, 5.11.
4-(3-Methoxyphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6g) Yield: 44%; mp: 115–117°C; IR (KBr), m
1
(cm-1): m = 1346 (C=S). H NMR (CDCl3), d (ppm): 5.12
1
(cm-1): 1344.43 (C=S); H NMR (CDCl3), d (ppm): 3.78
(s, 2H, N–CH2); 5.34 (s, 2H, O–CH2); 7.00–7.08 (m, aro-
matic, 3H); 7.20–7.23 (m, aromatic, 2H); 7.28–7.40 (m,
aromatic, 2H); 7.43–7.48 (m, aromatic, 2H). Anal. Calcd for
C15H13NOS: C, 70.56; H, 5.13; N, 5.49. Found: C, 70.83; H,
5.04; N, 5.28.
(OCH3), 5.11 (s, 2H, N–CH2); 5.33 (s, 2H, O–CH2);
6.75–6.82 (m, aromatic, 2H); 6.89–6.93 (m, aromatic, 1H);
7.00–7.08 (m, aromatic, 3H); 7.29–7.39 (m, aromatic, 2H).
Anal. Calcd for C16H15NO2S: C, 67.34; H, 5.30; N, 4.91;
Found: C, 67.38; H, 5.48; N, 4.84.
Spectroscopic and analytical data of compounds (6)
4-(4-Iodophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6h) Yield: 15%; mp: 163–164°C; IR (KBr), m
1
(cm-1): 1340.57 (C=S). H NMR (CDCl3), d (ppm): 5.09
4-(4-Dimethyaminophenyl)-4,5-dihydrobenzo[f][1,4]oxaze-
pin-3(2H)-thione (6a) Yield: 10%; mp: 195–196°C; IR
(KBr), m (cm-1): 1342.50 (C=S); 1H NMR (CDCl3), d
(ppm): 2.97 (s, 6H, N(CH3)2); 5.11 (s, 2H, N–CH2); 5.34
(s, 2H, O–CH2); 6.70–6.73 (m, aromatic, 2H); 6.98–7.08
(m, aromatic, 5H); 7.27–7.30 (m, aromatic, 1H). Anal.
Calcd for C17H18N2OS: C, 68.42; H, 6.08; N, 9.39. Found:
C, 68.13; H, 5.95; N, 9.29.
(s, 2H, N–CH2); 5.33 (s, 2H, O–CH2); 6.97–7.08 (m, aro-
matic, 5H); 7.30–7.34 (m, aromatic, 1H); 7.76–7.80 (m,
aromatic, 2H). Anal. Calcd for C15H12INOS: C, 47.26; H,
3.17; I, 33.29; N, 3.67; Found: C, 47.17; H, 3.27; N, 3.71.
4-(4-Chlorophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6i) Yield: 33%; mp: 138–139°C; IR (KBr), m
1
(cm-1): 1340.57 (C=S); H NMR (CDCl3), d (ppm): 5.09
4-(4-Methoxyphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6b) Yield: 18%; mp: 152–154°C; IR(KBr), m
(s, 2H, N–CH2); 5.33 (s, 2H, O–CH2); 7.00–7.08 (m, aro-
matic, 3H); 7.15–7.19 (m, aromatic, 2H); 7.30–7.40 (m,
aromatic, 1H); 7.41–7.44 (m, aromatic, 2H). Anal. Calcd
for C15H12ClNOS: C, 62.17; H, 4.17; N, 4.83; Found: C,
62.21; H, 4.35; N, 4.71.
1
(cm-1): 1346.36 (C=S); H NMR (CDCl3), d (ppm): 3.82
(s, 3H, O–CH3); 5.11 (s, 2H, N–CH2); 5.34 (s, 2H, O–
CH2); 6.94–7.07 (m, aromatic, 5H); 7.11–7.17 (m, aro-
matic, 2H); 7.28–7.34 (m, aromatic, 1H). Anal. Calcd for
C16H15NO2S: C, 67.34; H, 5.30; N, 4.91; Found: C, 67.17;
H, 5.14; N, 4.98.
4-(4-Bromophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6j) Yield: 32%; mp: 150–152°C; IR (KBr),m
1
(cm-1): 1340.57 (C=S). H NMR (CDCl3), d (ppm): 5.10
4-(4-Methylphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-thi-
one (6c) Yield: 26%; mp: 155–156°C; IR (KBr), m
(s, 2H, N–CH2); 5.33 (s, 2H, O–CH2); 6.98–7.05 (m, aro-
matic, 3H); 7.08–7.13 (m, aromatic, 2H); 7.26–7.34 (m,
aromatic, 1H); 7.51–7.60 (m, aromatic, 2H). Anal. Calcd
for C15H12BrNOS: C, 53.90; H, 3.62; N, 4.19; Found: C,
53.83; H, 3.81; N, 4.11.
1
(cm-1): 1346.36 (C=S); H NMR (CDCl3), d (ppm): 2.38
(s, 3H, CH3); 5.10 (s, 2H, N–CH2); 5.34 (s, 2H, O–CH2);
6.99–7.24 (m, aromatic, 5H); 7.27–7.34 (m, aromatic, 3H).
Anal. Calcd for C16H15NOS: C, 71.34; H, 5.61; N, 5.20;
Found: C, 71.25; H, 5.83; N, 5.22.
4-(3-Chlorophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6k) Yield: 10%; mp: 157–159°C; IR (KBr),m
4-(3-Methylphenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6d) Yield: 28%; mp: 129–130°C; IR (KBr),m
1
(cm-1): 1346.36 (C=S); H NMR (CDCl3), d (ppm): 5.10
1
(cm-1): 1344.43 (C=S); H NMR (CDCl3), d (ppm): 2.37
(CH3), 5.10 (s, 2H, N–CH2); 5.33 (s, 2H, O–CH2);
(s, 2H, N–CH2); 5.32 (s, 2H, O–CH2); 7.00–7.08 (m, aro-
matic, 3H); 7.11–7.15 (m, aromatic, 1H); 7.23–7.26 (m,
aromatic, 1H); 7.30–7.42 (m, aromatic, 3H). Anal. Calcd
6.98–7.20 (m, aromatic, 6H); 7.29–7.37 (m, aromatic, 2H).
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Med Chem Res (2011) 20:1170–1180
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Dilutions of the compounds
for C15H12ClNOS: C, 62.17; H, 4.17; N, 4.83; Found: C,
62.13; H, 4.44; N, 4.76.
For antibacterial activities, all of the dilutions were done
with Mueller–Hinton Broth (Oxoid) in the wells of mic-
rodilution plates. The concentrations of the tested com-
pounds were 1600, 800, 400, 200, 100, 50, 25, 12.5, 6.25,
3.12, 1.56, 0.78, 0.39, 0.19, 0.09, and 0.04 lg/ml. The
highest ampicillin and ciprofloxacin were used as a refer-
ence compounds which were obtained from the
manufacturers.
4-(3-Trifluoromethylphenyl)-4,5-dihydrobenzo[f][1,4]oxaze-
pin-3(2H)-thione (6l) Yield: 44%; mp: 112–113°C; IR
(KBr), m (cm-1): 1346.36 (C=S); 1H NMR (CDCl3), d
(ppm): 5.13 (s, 2H, N–CH2); 5.34 (s, 2H, O–CH2);
7.00–7.10 (m, aromatic, 3H); 7.34–7.37 (m, aromatic, 1H);
7.42–7.45 (m, aromatic, 1H); 7.52–7.56 (m, aromatic, 1H);
7.59–7.65 (m, aromatic, 2H). Anal. Calcd for
C16H12F3NOS: C, 59.43; H, 3.74; N, 4.33; Found: C,
59.43; H, 3.66; N, 4.27.
For antifungal activity, all the dilutions were done with
RPMI medium with L-glutamine buffered, pH 7, with
MOPS (Sigma) in the wells of microdilution plates. The
concentrations of 14 compounds tested are the same as
above. The highest fluconazole was used as a reference
compound which was also obtained from the manufacturer.
4-(3-Nitrophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6m) Yield: 16%; mp: 158–159°C; IR (KBr), m
1
(cm-1): 1346.36 (C=S); H NMR (CDCl3), d (ppm): 5.17
(s, 2H, N–CH2); 5.36 (s, 2H, O–CH2); 7.04–7.11 (m, aro-
matic, 3H); 7.33–7.36 (m, aromatic, 1H); 7.59–7.68 (m,
aromatic, 2H); 8.14–8.15 (m, aromatic, 1H); 8.22–8.26 (m,
aromatic, 1H). Anal. Calcd for C15H12N2O3S: C, 59.99; H,
4.03; N, 9.33; Found: C, 60.03; H, 3.52; N, 9.30.
Inoculum preparation
After diluting the compounds, standardized inoculum of
each bacterium (0.5 Mc Farland standard unit, 1 9 108
CFU/ml; (colony forming unit/ml) was prepared. Then, the
compounds were diluted once more (1/10), and final con-
centrations became 1 9 107 CFU/ml. Five microliters from
each dilution was placed into each well containing 100 ll
of dilutions of compounds so that each well contained
5 9 105 CFU/ml of inoculum. All the inoculated plates
were incubated at 35°C for 16–20 h. The lowest concen-
tration of compounds that prevents visible growth was
considered to be the minimal inhibitory concentration
(MIC). Ampicillin and ciprofloxacin are used as reference
antimicrobial reagents to compare their parameters with the
data that obtained from the method applied in this study
and to control the reliability of the results.
4-(4-Nitrophenyl)-4,5-dihydrobenzo[f][1,4]oxazepin-3(2H)-
thione (6n) Yield: 16%; mp: 182–184°C; IR (KBr), m
1
(cm-1): 1350.22 (C=S); H NMR (CDCl3), d (ppm): 5.14
(s, 2H, N–CH2); 5.35 (s, 2H, O–CH2); 7.04–7.10 (m, aro-
matic, 3H); 7.33–7.46 (m, aromatic, 3H); 8.31–8.34 (m,
aromatic, 2H). Anal. Calcd for C15H12N2O3S: C, 59.99; H,
4.03; N, 9.33; Found: C, 59.83; H, 4.19; N, 9.26.
Biological assays
The antibacterial activities of 14 compounds were deter-
mined using broth microdilution susceptibility test outlined
by the Clinical and Laboratory Standards Institute M7-
A713. Minimal inhibitory concentrations for each com-
pound were investigated against E. faecalis (ATCC
29212), S. aureus (ATCC 25983), E. coli (ATCC 25922),
and P. aeruginosa (ATCC 27853). For broth microdilution
procedures, sterile, disposable, multiwell microdilution
plates (96 U-shaped wells) were used. The stock solutions
were prepared in pure ethanol (Sigma). Ethanol had no
effect on the microorganisms in the concentrations studied.
The antifungal activities of the compounds were deter-
mined using broth microdilution susceptibility test outlined
by Clinical and Laboratory Standards Institute M27-A214.
Minimal inhibitory concentrations for each compound
were investigated against Candida albicans (ATCC
90028). For broth microdilution procedures, sterile, dis-
posable, multiwell microdilution plates (96 U-shaped
wells) were used. The stock solutions were prepared in
pure ethanol (Sigma) and again ethanol had no effect on the
microorganisms in the concentrations studied.
For antifungal activity, candida isolate were subcultured
in SDA plates, incubated at 35°C for 24–48 h prior to
antifungal susceptibility testing and passaged at least twice
to ensure purity and viability. An inoculum suspension was
prepared from individual five colonies (diameter 1 mm).
The suspension was adjusted to 0.5 MacFarland Standard
(1–5 9 106 CFU/ml) and further diluted 1/20 (1–5 9 105
CFU/ml) then 1/50 (0.5–2.5 9 105 CFU/ml) in RPMI
medium. 100 ll from each dilution was placed into each
well containing 100 ll of dilutions of compounds so that
each well contained 1 9 103 CFU/ml of inoculum. The
MIC plates were incubated at 37°C for 48 h. For the end
point was determined as the concentration producing
optically clear wells (MIC-0) compared with that of drug-
free growth control. Fluconazole was used as reference
antifungal reagent to compare its parameters with the data
that obtained from the method applied in this study and to
control the reliability of the results.
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Every experiment for the antibacterial and antifungal
assays was replicated twice. MIC values for the activities
are given in Tables 1 and 2, respectively.
Acknowledgment The authors thank Kocaeli University Research
Fund for financial support (Grant No. 2004/34).
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