Antimycobacterial 3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H)-ones and 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-dithiones substituted on phenyl and benzoxazine moiety in position 6

Article abstract of DOI:10.2478/s11696-011-0020-7

A series of forty-five derivatives of 3-phenyl-4-thioxo-2H-1,3-benzoxazine- 2(3H)-ones and forty-five derivatives of 3-phenyl-2H-1,3-benzoxazine-2,4(3H)- dithiones was synthesised. The compounds exhibited in-vitro activity against Mycobacterium tuberculos

Full text of DOI:10.2478/s11696-011-0020-7

Chemical Papers 65 (3) 352–366 (2011)
DOI: 10.2478/s11696-011-0020-7
ORIGINAL PAPER
Antimycobacterial
3-phenyl-4-thioxo-2 -1,3-benzoxazine-2(3 )-ones
and 3-phenyl-2 -1,3-benzoxazine-2,4(3 )-dithiones
substituted on phenyl and benzoxazine moiety in position 6
a
b
a
a
aEva Petrlíková*, Karel Waisser, Rafael Doležal, Pavel Holý, Jiří Gregor,
aJiří Kuneš, Jarmila Kaustová
c
aDepartment of Inorganic and Organic Chemistry, Charles University in Prague, Faculty of Pharmacy in Hradec Králové,
Heyrovského 1203, CZ-50003 Hradec Králové, Czech Republic
bFaculty of Science, University of Hradec Králové, Rokitanského 62, CZ-50003 Hradec Králové, Czech Republic
cRegional Institute of Public Health, Partyzánské náměstí 7, CZ-70200 Ostrava, Czech Republic
Received 13 October 2010; Revised 16 January 2011; Accepted 23 January 2011
A series of forty-five derivatives of 3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H)-ones and forty-
five derivatives of 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-dithiones was synthesised. The compounds
exhibited in-vitro activity against Mycobacterium tuberculosis, M. kansasii (two strains), and M.
avium. The most active derivatives were more active than isonicotinhydrazide (INH). The quan-
titative relationships between the structure and antimycobacterial activity were calculated. The
activity against M. tuberculosis increased with the lipophilicity of the substituents.
c
ꢀ 2011 Institute of Chemistry, Slovak Academy of Sciences
Keywords: benzoxazine, thioxo group, tuberculostatics, QSAR, antimycobacterial activity
Introduction
From the perspective of pharmaceutical treatment,
N-benzylsalicylamides, salicylanilides, and their cyclic
derivatives, benzoxazinediones, are promising classes
of compounds (Matyk et al., 2005; Nemeček et al.,
2009; Petrlíková et al., 2011, 2010; Waisser et al.,
2006). This study is oriented towards the derivatives
of benzoxazinediones in which one or both oxo groups
were replaced by the thioxo group. Since the com-
pounds are cyclic derivatives of salicylanilides, they
can be expected to serve as bacterial two-component
system inhibitors (Hlasta et al., 1998; Macielag et
al., 1998). Benzoxazine derivatives could also tar-
get the biosynthesis of menaquinone, a polyisopreny-
lated naphtoquinone, that plays an important role in
the mycobacterial electron transport chain (Li et al.,
2010). Both types of mechanism of action are very
promising since the consequent antibacterial effects
The prognosis that, following the millennium, tu-
berculosis would no longer occur in the developed
world was wrong (O’Brien & Nunn, 2001). The
emergence of multi-drug resistant (MDR-TB) and
extensively-drug resistant (XDR-TB) strains of My-
cobacterium tuberculosis is a serious problem and tu-
berculosis remains one of the leading infectious dis-
eases worldwide (Dye, 2009). This unfavourable state
is also being influenced by an increase in AIDS, which
is often accompanied by the mycobacterial diseases
and with the low standard of living of displaced per-
sons (Aaron et al., 2004; Naidoo et al., 2011). New
mycobacterial diseases are occurring which, until re-
cently, were considered non-transferable to humans
(Tortoli, 2009).
*Corresponding author, e-mail: eva.petrlikova@faf.cuni.cz

E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
353
are probably different from the effects of other antibac-
terial drugs (Schroeder et al., 2002; van den Boogaard
et al., 2009).
300 MHz for 1H NMR and 75 MHz for 13C NMR.
Chemical shifts were recorded as δ values and indi-
rectly referenced to tetramethylsilane via the solvent
1
signal (DMSO) (2.5 for H or 39.5 for 13C).
Theoretical
General procedure for the preparation of
3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H)-
ones (I–V) and 3-phenyl-2H-1,3-benzoxazine-
2,4(3H)-dithiones (VI–X)
Regressions were calculated using Microsoft Ex-
cel Multireg programs. The values of the substituent
π and σ constants were taken from the literature
(Hansch & Leo, 1979). The stability of the QSAR
models was evaluated by cross-validation (leave-one-
out procedure) (Gupta et al., 2009; Golbraikh & Trop-
sha, 2002) in Matlab 7.0 program. The Free–Wilson
method (Free & Wilson, 1964) modified by Fujita–Ban
(Fujita & Ban, 1971) was used to investigate the ac-
tivity contribution in the case of sulphur derivatives.
Since the MIC values after 14 d and 21 d incubation
correlated with each other, only the MICs after 14 d
evaluation were taken for the calculations.
The derivatives of 3-phenyl-2H-1,3-benzoxazine-
2,4(3H)-diones (4 mmol) underwent fusion with P4S10
(10 mmol) for 45 min (180–210◦C). After cooling
to room temperature, a 10 % potassium carbon-
ate solution was poured into the reaction mixture;
the crude product was removed by filtration, and
dissolved in toluene (p.a., a maximum of 40 mL).
Column chromatography using silica gel provided
substituted derivatives of 3-phenyl-4-thioxo-2H-1,3-
benzoxazine-2(3H)-one (I–V) and substituted deriva-
tives of 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-dithione
(VI–X) as orange-yellow and red solids, respectively.
Recrystallisation from ethanol was necessary.
Experimental
Materials and methods
The melting points were determined using the
Kofler apparatus (C. Reichert, Vienna, Austria) and
the temperature data were corrected. The samples for
analyses and antimycobacterial tests were dried over
P2O5 at 61◦C and 66 Pa for 24 h. The elemental anal-
yses (C, H, N, S) were performed on a CHNS-O CE el-
emental analyzer (Fisions EA 1110, Milan, Italy) and
were within 0.4 % of the theoretical values. The IR
spectra of KBr pellets were measured on a Nicolet Im-
pact 400 apparatus (Nicolet, Madison, WI, USA); the
wavenumbers are given in cm−1. The purity of the
compounds was verified by TLC on silica gel plates
pre-coated with a fluorescent indicator Silufol UV 254
+ 366 (Kavalier Votice, Czech Republic) and hexane–
acetone mixture (ϕr = 3 : 1) as the mobile phase. The
1H NMR and 13C NMR spectra of new compounds
were recorded in DMSO-d6 solutions at ambient tem-
perature on a Varian Mercury-Vx BB 300 spectrom-
eter (Varian Inc., Palo Alto, CA, USA) operating at
Antimycobacterial susceptibility testing
For the in-vitro evaluation of the antimycobacte-
rial activity of the substances, the following strains
were used: M. tuberculosis CNCTC My 331/88 (iden-
tical with H37RV and ATCC 27294), M. kansasii
CNCTC My 235/80 (identical with ATCC 12 478),
M. avium CNCTC My 330/88 (identical with ATCC
25291), obtained from the Czech National Collec-
tion of Type Cultures (CNCTC), National Institute
of Public Health, Prague, and a clinical isolate ofM.
kansasii 6509/96. The antimycobacterial activity of
the compounds was determined in the Šula semisyn-
thetic medium (SEVAC, Prague). In order to con-
trol the sterility of the inoculum and its growth, a
Petri dish containing the Lo¨wenstein–Jensen medium
was inoculated with each strain. The compounds were
added to the medium dissolved in DMSO. The final
concentrations were 1000 µmol L−1, 500 µmol L−1
,
O
O
N
N
R1
S
V
O
O
O
R2
P2S5
I
N
R1
O
S
R2
R1
S
R2
VI
X
Fig. 1. Synthesis of 3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H )-ones and 3-phenyl-2H-1,3-benzoxazine-2,4(3H )-dithiones.

354
E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Table 1. Overview and characteristics of 3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H )-ones
wi(calc.)/mass %
wi(found)/mass %
Yield
M.p.
ν˜ (CO)
Comp.
R1
R2
Formula
Mr
C
H
N
S
%
◦C
cm−1
1764
Ia
Ib
Ic
Id
Ie
If
Br
Br
4-H
4-CH3
4-Cl
C14H8BrNO2S
C15H10BrNO2S
334.2
348.2
50.32
50.30
51.74
51.22
45.61
45.31
45.61
45.29
41.72
41.42
40.71
40.43
47.75
47.56
47.75
47.67
44.80
44.63
66.89
66.51
2.41
2.45
2.89
2.62
1.91
1.99
1.91
2.01
1.50
1.85
1.71
1.65
2.00
2.32
2.00
1.88
1.75
2.01
4.12
4,44
4.19
4.12
4.02
3.98
3.80
3.53
3.80
3.62
3.47
3.32
3.39
3.15
3.98
3.63
3.98
3.68
3.48
3.16
5.20
4.98
9.59
9.32
9.21
9.11
8.70
8.52
8.70
8.35
7.96
7.57
7.76
7.51
9.10
8.83
9.10
8.79
7.97
7.68
11.91
11.63
38
240–242
36
31
33
30
37
30
39
35
34
259–260.5
283–284a
199–200
220–222
260–262b
290–291
227–228
247–248
217–219
1759
1759
1759
1961
1771
1763
1764
1751
1756
Br
C14H7BrClNO2S 368.6
C14H7BrClNO2S 368.6
Br
3-Cl
Br
3,4-Cl2 C14H6BrCl2NO2S 403.1
Br
4-Br
4-F
3-F
C14H7Br2NO2S
413.1
Ig
Ih
Ii
Br
C14H7BrFNO2S 352.2
C14H7BrFNO2S 352.2
Br
Br
4-CF3 C15H7BrF3NO2S 402.2
IIa
CH3
4-H
C15H11NO2S
269.3
IIb
IIc
IId
IIe
CH3
CH3
CH3
4-CH3
4-Cl
C16H13NO2S
C15H10ClNO2S
C15H10ClNO2S
C15H9Cl2NO2S
283.3
303.8
303.8
338.2
67.82
67.53
59.31
58.95
59.31
59.63
53.27
53.52
4.62
4.84
3.32
3.65
3.32
3.70
2.68
2,87
4.94
4.75
4.61
4.43
4.61
4.68
4.14
4.53
11.32
11.21
10.56
10.37
10.56
10.25
9.48
30
32
31
37
218–219.5
201–203
175–177
185–188
1755
1755
1756
1754
3-Cl
CH3 3,4-Cl2
9.21
IIf
IIg
IIh
IIi
CH3
CH3
4-Br
4-F
C15H10BrNO2S
C15H10FNO2S
C15H10FNO2S
C16H10F3NO2S
C15H11NO3S
348.2
287.3
287.3
337.3
285.3
51.74
51.52
62.71
62.43
62.71
62.64
56.97
56.64
63.14
62.85
2.89
2.68
3.51
3.79
3.51
3.32
2.99
3.26
3.89
4.01
4.02
5.21
4.88
4.52
4.88
4.65
4.15
3.95
4.91
4.73
9.21
8.94
36
31
38
35
32
207–209
206–207
189–199
237–238
233–235
1759
1757
1758
1760
1754
11.16
10.83
11.16
11.45
9.51
CH3
3-F
CH3
4-CF3
4-H
9.28
IIIa
CH3O
11.24
11,55
IIIb
IIIc
IIId
IIIe
IIIf
IIIg
IIIh
IIIi
IVa
IVb
CH3O 4-CH3
C16H13NO3S
C15H10ClNO3S
C15H10ClNO3S
C15H9Cl2NO3S
C15H10BrNO3S
C15H10FNO3S
C15H10FNO3S
C16H10F3NO3S
C14H8FNO2S
C15H10NO2S
299.3
319.8
319.8
354.2
364.2
303.3
303.3
353.3
273.3
287.3
64.20
63.98
56.34
56.01
56.34
56.22
50.86
51.08
49.47
49.76
59.40
59.21
59.40
59.01
54.39
54.22
61.53
61.34
62.71
62.59
4.38
4.56
3.15
3.36
3.15
2.99
2.56
2.83
2.77
3.05
3.32
3.64
3.32
3.72
2.85
3.24
2.95
3.28
3.51
3.83
4.68
4.52
4.38
3.99
4.38
4.10
3.95
3.74
3.85
3.68
4.62
4.28
4.62
4.39
3.96
4.01
5.13
4.78
4.88
4.64
10.71
10.49
10.03
9.85
31
35
31
38
36
32
36
34
30
32
194–195.5
227–228
234–235
211–213
219–220
223–224
245–246
202
1757
1755
1755
1754
1750
1759
1763
1751
1762
1756
CH3O
CH3O
4-Cl
3-Cl
10.03
9.99
CH3O 3,4-Cl2
9.05
8.87
CH3O
CH3O
CH3O
4-Br
4-F
3-F
8.80
8.45
10.57
10.26
10.57
10.39
9.08
CH3O 4-CF3
8.86
F
F
H
11.73
11.93
11.16
10.98
228–229
211–213
4-CH3

E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
355
Table 1. (continued)
wi(calc.)/mass %
wi(found)/mass %
Yield
M.p.
ν˜ (CO)
Comp. R1
R2
Formula
Mr
C
H
N
S
%
33
34
30
31
32
31
33
34
30
32
◦C
cm−1
1756
1756
1756
1755
1768
1755
1765
1764
1766
1754
IVc
IVd
IVe
IVf
IVg
IVh
IVi
Vg
F
F
4-Cl
3-Cl
C14H7ClFNO2S 307.7
C14H7ClFNO2S 307.7
54.64
54.37
54.64
54.24
49.14
48.86
47.75
42.56
57.73
58.01
57.73
57.56
52.79
53.05
54.64
54.48
54.64
54.28
50.36
50.16
2.29
2.48
2.29
2.52
1.77
2.01
2.00
2.26
2.42
2.73
2.42
2.68
2.07
2.38
2.29
2.56
2.29
2.63
1.97
2.35
4.55
4.37
4.55
4.43
4.09
3.98
3.98
3.69
4.81
4.32
4.81
4.46
4.10
3.85
4.55
4.24
4.55
4.19
3.92
3.66
10.42
10.17
10.42
10.36
9.37
211–213
184–185
203–205
230–231
175–178
172–174
207–208
276–277
209–210
235–236
F
3,4-Cl2 C14H6Cl2FNO2S 347.2
8.09
F
4-Br
4-F
C14H7BrFNO2S 352.2
9.10
8.85
F
C14H7F2NO2S
C14H7F2NO2S
C15H7F4NO2S
291.3
291.3
341.3
11.01
10.83
11.01
10.69
9.40
F
3-F
F
4-CF3
4-F
9.73
Cl
Cl
Cl
C14H7ClFNO2S 307.7
C14H7ClFNO2S 307.7
10.42
10.27
10.42
10.18
8.96
Vh
3-F
Vi
4-CF3 C15H7ClF3NO2S 352.7
8.68
a) 280–281◦C (Wagner et al., 1966); b) 256–258◦C (Wagner et al., 1966).
Table 2. Overview and characteristics of 3-phenyl-2H-1,3-benzoxazine-2,4(3H )-dithiones
wi(calc.)/mass %
wi(found)/mass %
Yield
M.p.
Comp.
R1
R2
Formula
Mr
C
H
N
S
%
32
31
33
31
30
33
32
35
32
30
32
33
31
32
◦C
VIa
VIb
VIc
VId
VIe
VIf
Br
Br
4-H
4-CH3
4-Cl
C14H8BrNOS2
C15H10BrNOS2
C15H7BrClNOS2
C15H7BrClNOS2
C14H6BrCl2NOS2
C14H7Br2NOS2
C15H7BrFNOS2
C14H7BrFNOS2
C15H7BrF3NOS2
C16H13NOS2
350.3
364.3
384.7
384.7
419.1
429.2
368.2
368.2
418.3
285.4
299.4
319.8
319.8
354.3
48.01
47.86
49.46
49.23
43.71
43.48
43.71
43.51
40.12
39.87
39.18
38.96
45.66
45.83
45.66
45.24
43.07
42.89
63.13
62.84
64.18
63.82
56.33
55.99
56.33
56.20
50.85
51.17
2.30
2.45
2.77
2.96
1.83
2.03
1.83
1.99
1.44
1.79
1.64
1.91
1.92
2.30
1.92
2.14
1.69
201
4.00
3.82
3.85
3.62
3.64
3.23
3.64
3.46
3.34
2.98
3.26
3.01
3.80
3.64
3.80
3.51
3.35
3.71
4.91
5.21
4.68
5.01
4.38
4.01
4.38
4.25
3.95
3.95
18.31
244–245
246–248
256–258
190–192
203–205
248–251
224–226
231–232
182–184
198–199
225–227
192–193
185–186
175–176
18.26
17.60
17.32
16.67
16.29
16.67
16.55
15.30
14.98
14.94
15.02
17.42
17.67
17.42
17.23
15.33
15.58
22.47
22.28
21.42
21.26
20.05
19.76
20.05
20.36
18.10
17.79
Br
Br
3-Cl
Br
3,4-Cl2
4-Br
Br
VIg
VIh
VIi
Br
4-F
Br
3-F
Br
4-CF3
4-H
VIIa
VIIb
VIIc
VIId
VIIe
CH3
CH3
CH3
CH3
CH3
3.89
3.73
4.38
4.32
3.15
2.87
3.15
3.38
2.56
2.83
4-CH3
4-Cl
C16H13NOS2
C15H10ClNOS2
C15H10ClNOS2
C15H9Cl2NOS2
3-Cl
3,4-Cl2

356
E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Table 2. (continued)
wi(calc.)/mass %
wi(found)/mass %
Yield
M.p.
Comp.
R1
R2
Formula
Mr
C
H
N
S
%
30
33
31
31
32
30
32
33
30
31
32
30
33
30
32
31
33
32
32
30
33
31
32
31
30
◦C
VIIf
VIIg
VIIh
VIIi
VIIIa
VIIIb
VIIIc
VIIId
VIIIe
VIIIf
VIIIg
VIIIh
VIIIi
IXa
CH3
CH3
4-Br
4-F
C15H10CBrNOS2
C15H10FNOS2
C15H10FNOS2
C16H10F3NOS2
C15H11NO2S3
364.3
303.4
303.4
353.3
301.4
315.4
335.8
335.8
370.3
380.3
319.4
319.4
369.4
289.4
303.4
323.8
323.8
358.2
368.2
307.3
307.3
357.4
323.8
323.8
373.8
49.46
49.17
59.39
59.18
59.39
59.17
54.38
54.23
59.78
59.66
60.93
60.68
53.65
53.28
53.65
53.36
48.66
48.24
47.38
47.16
56.41
56.23
56.41
56.28
52.03
51.96
58.11
57.86
59.39
59.19
51.93
51.77
51.93
51.68
46.94
46.62
45.66
46.38
54.71
54.83
54.71
54.64
50.42
50.77
51.93
51.58
51.93
51.88
48.20
47.85
2.77
3.01
3.32
3.17
3.32
3.63
2.85
3.12
3.68
3.97
4.15
4.48
3.00
3.38
3.00
3.27
2.45
2.63
2.65
2.87
3.16
3.24
3.16
3.36
2.73
2.98
2.79
2.93
3.32
3.54
2.18
2.34
2.18
2.42
1.69
2.01
1.92
2.37
2.30
2.44
2.30
2.63
1.97
2.23
2.18
2.14
2.18
1.96
1.89
2.17
3.85
3.63
4.62
4.24
4.62
4.27
3.96
3.73
4.65
4.27
4.44
4.23
4.17
3.82
4.17
3.96
3.78
3.63
3.68
3.37
4.39
4.16
4.39
4.23
3.79
3.56
4.84
4.53
4.62
4.36
4.33
4.14
4.33
4.57
3.91
3.58
3.80
3.52
4.56
4.16
4.56
4.27
3.92
3.68
4.33
4.22
4.33
4.61
4.28
3.91
17.60
17.43
21.14
20.89
21.14
20.78
18.15
17.96
21.28
20.12
20.33
20.17
19.10
18.78
19.10
18.84
17.32
17.12
16.86
16.73
20.08
19.89
20.08
20.15
17.36
17.15
22.16
21.96
21.14
20.83
19.81
19.76
19.81
19.53
17.90
17.65
17.42
17.27
20.87
20.93
20.87
20.74
17.95
17.86
19.81
19.99
19.81
20.02
17.16
17.38
218–219
184–185
193–194
194–195
259–260
190–191
181–183
219–220
170–172
171–173
211–212
278–279
195–196
166–167
187–189
166–168
166–168
162–164
177–178
180–182
189–191
178–179
203–204
213
CH3
3-F
CH3
4-CF3
4-H
CH3O
CH3O
CH3O
CH3O
4-CH3
4-Cl
3-Cl
C16H13NO2S2
C15H10ClNO2S2
C15H10ClNO2S2
C15H9Cl2NO2S2
C15H10BrNO2S2
C15H10FNO2S2
C15H10FNO2S2
C16H10F3NO1S2
C14H8FNO2S2
C15H10NOS2
CH3O 3,4-Cl2
CH3O
4-Br
4-F
CH3O
CH3O
3-F
CH3O
F
4-CF3
H
IXb
F
4-CH3
4-Cl
3-Cl
3,4-Cl2
4-Br
4-F
IXc
F
C14H7ClFNOS2
C14H7ClFNOS2
C14H6Cl2FNOS2
C14H7BrFNOS2
IXd
F
IXe
F
IXf
F
IXg
F
C14
C14
H
H
7F2NOS2
7F2NOS2
IXh
F
3-F
IXi
F
4-CF3
4-F
C15H7F4NOS2
C14H7ClFNOS2
C14H7ClFNOS2
C15H7ClF3NOS2
Xg
Cl
Cl
Cl
Xh
3-F
Xi
4-CF3
175–176
250 µmol L−1, 125 µmol L−1, 62.5 µmol L−1, 32 µmol
L
L
tively. The tests were repeated three times.
−1, 16 µmol L−1, 8 µmol L−1, 4 µmol L−1, 2 µmol
−1, 1 µmol L−1, and 0.5 µmol L−1. The minimum in-
Results and discussion
hibitory concentrations (MIC) were determined after
incubation at 37◦C for 14 days and 21 days, respec-
3-Phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H)-ones

E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Table 3. NMR spectral data of newly prepared compounds
357
Comp.
NMR spectral dataa
Ia
1H NMR (300 MHz, CDCl3), δ: 8.54 (d, 1H, J = 2.35 Hz, H-5), 7.80 (dd, 1H, J = 8.66 Hz, J = 2.35 Hz, H-7),
7.61–7.50 (m, 3H, H-3ꢀ, H-4ꢀ, H-5ꢀ), 7.28–7.19 (m, 3-H, H-8, H-2ꢀ, H-6ꢀ)
13C NMR (75 MHz, CDCl3), δ: 189.9, 148.3, 144.5, 138.8, 138.4, 134.3, 129.9, 129.3, 127.7, 122.0, 118.8, 118.4
Ib
Ic
Id
Ie
1H NMR (300 MHz, CDCl3), δ: 8.54 (d, 1H, J = 2.45 Hz, H-5), 7.80 (dd, 1H, J = 8.52 Hz, J = 2.45 Hz, H-7),
7.40–7.34 (m, AAꢀ, BBꢀ, 2-H, H-2ꢀ, H-6ꢀ), 7.20 (d, 1H, J = 8.52 Hz, H-8), 7.16–7.11 (m, AAꢀ, BBꢀ, 2-H, H-3ꢀ, H-5ꢀ),
2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 190.0, 148.2, 144.6, 139.5, 138.4, 136.2, 134.3, 130.6, 127.3, 122.0, 118.8, 118.4, 21.5
1H NMR (300 MHz, CDCl3), δ: 8.53 (d, 1H, J = 2.40 Hz, H-5), 7.81 (dd, 1H, J = 8.82 Hz, J = 2.40 Hz, H-7),
7.56–7.49 (m, AAꢀ, BBꢀ, 2-H, H-2ꢀ, H-6ꢀ), 7.21 (d, overlapped, 1H, J = 8.82 Hz, H-8), 7.24–7.16 (m, AAꢀ, BBꢀ, 2-H,
H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, CDCl3), δ: 189.7, 148.2, 144.4, 138.6, 137.1, 135.4, 134.3, 130.2, 129.2, 121.9, 119.0, 118.5
1H NMR (300 MHz, CDCl3), δ: 8.51 (d, 1H, J = 2.41 Hz, H-5), 7.81 (dd, 1H, J = 8.83 Hz, J = 2.41 Hz, H-7),
7.51–7.47 (m, 2-H, H-2ꢀ, H-4ꢀ), 7.28–7.25 (m, 1-H, H-6ꢀ), 7.21 (d, 1H, J = 8.83 Hz, H-8), 7.18–7.12 (m, 1-H, H-5ꢀ)
13C NMR (75 MHz, CDCl3), δ: 189.5, 148.2, 144.2, 139.5, 138.6, 135.3, 134.2, 130.8, 129.7, 128.3, 126.2, 121.9,
119.0, 118.4
1H NMR (300 MHz, CDCl3), δ: 8.51 (d, 1H, J = 2.48 Hz, H-5), 7.82 (dd, 1H, J = 8.71 Hz, J = 2.48 Hz, H-7), 7.63
(d, 1H, J = 8.51 Hz, H-5ꢀ), 7.37 (d, 1H, J = 2.42 Hz, H-2ꢀ), 7.21 (d, 1H, J = 8.71 Hz, H-8), 7.11 (dd, 1H, J = 8.51
Hz, J = 2.42 Hz, H-6ꢀ)
13C NMR (75 MHz, CDCl3), δ: 189.4, 148.1, 144.2, 138.8, 137.5, 134.2, 133.9, 133.9, 131.5, 130.1, 127.4, 121.8,
119.1, 118.5
If
1H NMR (300 MHz, CDCl3), δ: 8.52 (d, 1H, J = 2.42 Hz, H-5), 7.81 (dd, 1H, J = 8.80 Hz, J = 2.42 Hz, H-7),
7.71–7.65 (m, AAꢀ, BBꢀ, 2-H, H-2ꢀ, H-6ꢀ), 7.21 (d, 1H, J = 8.80 Hz, H-8), 7.16–7.09 (m, AAꢀ, BBꢀ, 2-H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, CDCl3), δ: 189.6, 148.2, 144.3, 138.6, 137.6, 134.2, 133.2, 129.5, 123.5, 121.9, 119.0, 118.5
Ig
1H NMR (300 MHz, CDCl3), δ: 8.53 (d, 1H, J = 2.47 Hz, H-5), 7.81 (dd, 1H, J = 8.79 Hz, J = 2.47 Hz, H-7),
7.25–7.19 (m, 4-H, H-2ꢀ, H-3ꢀ, H-5ꢀ, H-6ꢀ), 7.21 (d, overlapped, 1H, J = 8.79 Hz, H-8)
13C NMR (75 MHz, CDCl3), δ: 190.2, 162.7 (d, J = 249.9 Hz), 148.4, 144.7, 138.7, 134.7 (d, J = 3.4 Hz), 134.4,
129.8 (d, J = 8.9 Hz), 122.1, 119.0, 118.5, 117.1 (d, J = 23.2 Hz)
Ih
1H NMR (300 MHz, CDCl3), δ: 8.52 (d, 1H, J = 2.47 Hz, H-5), 7.81 (dd, 1H, J = 8.79 Hz, J = 2.47 Hz, H-7),
7.58–7.49 (m, 1H, H-6ꢀ), 7.27–7.21 (m, 1H, H-2ꢀ), 7.21 (d, overlapped, 1H, J = 8.79 Hz, H-8), 7.09–7.03 (m, 1H,
H-5ꢀ), 7.00 (dt, 1H, J = 8.79 Hz, J = 2.20 Hz, H-4ꢀ)
13C NMR (75 MHz, CDCl3), δ: 189.7, 163.3 (d, J = 248.8 Hz), 148.3, 144.4, 139.8 (d, J = 10.0 Hz), 138.7, 134.3,
131.1 (J = 8.9 Hz), 123.8 (d, J = 3.47 Hz), 122.0, 119.1, 118.5, 116.7 (d, J = 20.9 Hz), 115.9 (d, J = 23.8 Hz)
Ii
1H NMR (300 MHz, DMSO-d6), δ: 8.36 (d, 1H, J = 2.47 Hz, H-5), 8.06 (dd, 1H, J = 8.79 Hz, J = 2.47 Hz, H-7),
7.96–7.90 (m, AAꢀ, BBꢀ, 2-H, H-3ꢀ, H-5ꢀ), 7.69–7.62 (m, AAꢀ, BBꢀ, 2-H, H-2ꢀ, H-6ꢀ), 7.52 (d, 1H, J = 8.79 Hz, H-8)
13C NMR (75 MHz, DMSO-d6), δ: 191.2, 149.2, 144.3, 143.5, 138.9, 132.8, 129.7, 129.5 (q, J = 32.1 Hz), 127.0 (q,
J = 3.7 Hz), 124.2 (q, J = 272.6 Hz), 122.5, 119.5, 117.6
IIa
IIb
1H NMR (300 MHz, (CD3)2CO), δ: 8.22 (d, 1H, J = 1.78 Hz, H-5), 7.60–7.47 (m, 4H, H-7, H-3ꢀ, H-4ꢀ, H-5ꢀ),
7.29–7.24 (m, 2H, H-2ꢀ, H-6ꢀ), 7.20 (d, 1H, J = 8.24 Hz, H-8), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, (CD3)2CO), δ: 192.1, 147.6, 145.3, 139.2, 137.1, 135.9, 131.8, 129.8, 129.2, 127.9, 120.5, 116.3,
20.8
1H NMR (300 MHz, (CD3)2CO), δ: 8.22 (d, 1H, J = 1.92 Hz, H-5), 7.52 (dd, 1H, J = 8.24 Hz, J = 1.92 Hz, H-7),
7.40–7.33 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.20 (d, 1H, J = 8.24 Hz, H-8), 7.17–7.12 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ),
2.45 (bs, 6H, CH3)
13C NMR (75 MHz, (CD3)2CO), δ: 192.2, 147.6, 145.4, 139.3, 137.0, 136.6, 135.9, 131.8, 130.6, 127.5, 120.5, 116.3,
21.4, 20.8
IIc
IId
1H NMR (300 MHz, (CD3)2CO), δ: 8.20 (d, 1H, J = 1.62 Hz, H-5), 7.57–7.49 (m, 3H, H-7, H-2ꢀ, H-6ꢀ), 7.24–7.16
(m, 3H, H-8, H-3ꢀ, H-5ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, (CD3)2CO), δ: 191.9, 147.5, 145.1, 137.5, 137.2, 136.1, 135.2, 131.7, 130.1, 129.4, 120.4, 116.3,
20.8
1H NMR (300 MHz, (CD3)2CO), δ: 8.21–8.18 (m, 1H, H-5), 7.54 (ddd, 1H, J = 8.24 Hz, J = 2.20 Hz, J = 0.57 Hz,
H-7), 7.50–7.45 (m, 2H, H-2ꢀ, H-6ꢀ), 7.29–7.27 (m, 1H, H-5ꢀ), 7.20 (d, overlapped, 1H, J = 8.24 Hz, H-8), 7.20–7.14
(m, 1H, H-4ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, (CD3)2CO), δ: 191.7, 147.5, 145.0, 140.0, 137.3, 136.1, 135.3, 131.7, 130.7, 129.6, 128.5, 126.4,
120.3, 116.3, 20.8
IIe
1H NMR (300 MHz, (CD3)2CO), δ: 8.20–8.17 (m, 1H, H-5), 7.62 (d, 1H, J = 8.52 Hz, H-5ꢀ), 7.54 (dd, 1H, J = 8.52
Hz, J = 2.20 Hz, H-7), 7.38 (d, 1H, J = 2.47 Hz, H-2ꢀ), 7.20 (d, 1H, J = 8.52 Hz, H-8), 7.12 (dd, 1H, J = 8.52 Hz,
J = 2.47 Hz, H-6ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, (CD3)2CO), δ: 191.6, 147.5, 145.0, 138.0, 137.4, 136.2, 133.8, 133.8, 131.7, 131.5, 130.3, 127.6,
120.3, 116.4, 20.8

358
E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Table 3. (continued)
Comp.
NMR spectral dataa
IIf
IIg
IIh
1H NMR (300 MHz, (CD3)2CO), δ: 8.21–8.19 (m, 1H, H-5), 7.71–7.64 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.54 (m, 1H,
H-7), 7.20 (d, 1H, J = 8.24 Hz, H-8), 7.17–7.11 (m, AAꢀ, BBꢀ, 2H, H3ꢀ, H5ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, (CD3)2CO), δ: 191.8, 147.5, 145.1, 138.1, 137.3, 136.1, 133.1, 131.7, 129.7, 123.3, 120.4, 116.3,
20.8
1H NMR (300 MHz, (CDCl3), δ: 8.20 (d, 1H, J = 2.20 Hz, H-5), 7.53 (dd, 1H, J = 8.24 Hz, J = 2.20 Hz, H-7),
7.25–7.22 (m, 4H, H-2ꢀ H-3ꢀ, H-5ꢀ, H-6ꢀ), 7.20 (d, 1H, J = 8.24 Hz, H-8), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 192.1, 162.6 (d, J = 249.4 Hz), 147.5, 145.3, 137.2, 136.0, 135.0 (d, J = 3.5 Hz),
131.8, 129.9 (d, J = 8.8 Hz), 120.4, 117.0 (d, J = 23.2 Hz), 116.3, 20.8
1H NMR (300 MHz, (CDCl3), δ: 8.22–8.19 (m, 1H, H-5), 7.57–7.48 (m, 2H, H-7, H-6ꢀ), 7.25–7.17 (m, 1H, H-2ꢀ),
7.20 (d, overlapped, 1H, J = 8.52 Hz, H-8), 7.09–7.04 (m, 1H, H-5ꢀ), 7.01 (dd, 1H, J = 8.79 Hz, J = 2.19 Hz, H-4ꢀ),
2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 191.7, 163.2 (d, J = 248.8 Hz), 147.5, 145.0, 140.2 (d, J = 10.3 Hz), 137.2, 136.1,
131.7, 131.0 (d, J = 8.9 Hz), 123.9 (d, J = 3.4 Hz), 120.4, 116.5 (d, J = 20.9 Hz), 115.9 (d, J = 23.8 Hz), 20.8
IIi
1H NMR (300 MHz, DMSO-d6), δ: 8.13–8.08 (m, 1H, H-5), 7.96–7.88 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 7.70–7.62 (m,
AAꢀ, BBꢀ overlapped, 2H, H-2ꢀ, H-6ꢀ), 7.70 (dd, overlapped, 1H, J = 8.52 Hz, J = 2.20 Hz, H-7), 7.40 (d, 1H, J =
8.51 Hz, H-8), 2.41 (s, 3H, CH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.7, 148.1, 144.8, 143.8, 137.6, 135.5, 130.8, 129.8, 129.3 (q, J = 31.8 Hz),
126.8 (q, J = 3.8 Hz), 124.2 (q, J = 272.6 Hz), 120.6, 116.7, 20.5
IIIa
IIIb
1H NMR (300 MHz, CDCl3), δ: 7.83 (d, 1H, J = 2.74 Hz, H-5), 7.64–7.45 (m, 3H, H-7, H-2ꢀ, H-6ꢀ), 7.37–7.19 (m,
4H, H-8, H-3ꢀ, H-4ꢀ, H-5ꢀ), 3.90 (s, 3H, OCH3)
13C NMR (75 MHz, CDCl3), δ: 191.7, 157.1, 143.9, 139.3, 130.0, 129.9, 129.3, 127.9, 125.3, 125.0, 117.9, 112.5, 56.0
1H NMR (300 MHz, DMSO-d6), δ: 7.74–7.70 (m, 1H, H-5), 7.56–7.35 (m, 3H, H-7, H-2ꢀ, H-6ꢀ), 7.33–7.20 (m, 3H,
H-8, H-3ꢀ, H-5ꢀ), 3.83 (s, 3H, OCH3), 2.36 (s, 3H, CH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.5, 156.5, 144.3, 138.2, 137.8, 130.0, 129.5, 128.1, 124.4, 121.3, 118.3, 112.6,
56.1, 21.0
IIIc
IIId
IIIe
IIIf
IIIg
IIIh
IIIi
IVa
1H NMR (300 MHz, DMSO-d6), δ: 7.72–7.70 (m, 1H, H5), 7.64–7.55 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.49–7.39 (m,
4H, H-7, H-8, H-3ꢀ, H-5ꢀ), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.4, 156.5, 144.8, 144.3, 139.2, 133.5, 130.5, 129.7, 124.6, 121.9, 118.3, 112.5,
56.1
1H NMR (300 MHz, DMSO-d6), δ: 7.73–7.69 (m, 1H, H5), 7.59–7.50 (m, 3H, H-7, H-2ꢀ, H-6ꢀ), 7.48–7.38 (m, 3H,
H-8, H-4ꢀ, H-5ꢀ), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.3, 156.6, 144.8, 144.2, 141.5, 133.5, 131.2, 129.0, 128.6, 127.6, 124.6, 121.3,
118.3, 112.5, 56.1
1H NMR (300 MHz, DMSO-d6), δ: 7.83 (d, 1H, J = 8.52 Hz, H-5ꢀ), 7.79 (d, 1H, J = 2.20 Hz, H-5), 7.72–7.70 (m,
1H, H-2ꢀ), 7.50–7.43 (m, 3H, H-7, H-8, H-6ꢀ), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.3, 156.6, 144.7, 144.2, 140.0, 131.8, 131.8, 131.7, 130.8, 129.4, 124.7, 121.2,
118.4, 112.4, 56.1
1H NMR (300 MHz, DMSO-d6), δ: 7.75–7.69 (m, 3H, H-5, H-2ꢀ, H-6ꢀ), 7.47–7.44 (m, 2H, H-7, H-8), 7.40–7.34 (m,
2H, H-3ꢀ, H-5ꢀ), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.4, 156.5, 144.8, 144.3, 139.6, 132.7, 130.8, 124.6, 122.1, 121.3, 118.3, 112.5,
56.1
1H NMR (300 MHz, DMSO-d6), δ: 7.71 (dd, 1H, J = 2.47 Hz, J = 0.82 Hz, H-5), 7.50–7.41 (m, 4H, H-7, H-8, H-2ꢀ,
H-6ꢀ), 7.40–7.31 (m, 2H, H-3ꢀ, H-5ꢀ), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 193.1, 162.4 (d, J = 245.1 Hz), 157.0, 145.4, 144.7, 137.0 (d, J = 3.2 Hz), 131.2
(d, J = 8.9 Hz), 125.0, 121.8, 118.9, 117.0 (d, J = 23.2 Hz), 113.1, 56.6
1H NMR (300 MHz, DMSO-d6), δ: 7.73–7.70 (m, 1H, H-5), 7.62–7.42 (m, 3H, H-7, H-2ꢀ, H-6ꢀ), 7.38–7.24 (m, 3H,
H-8, H-4ꢀ, H-5ꢀ), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.3, 162.6 (d, J = 244.0 Hz), 156.6, 144.7, 144.2, 141.5 (d, J = 10.9 Hz), 131.2
(d, J = 8.9 Hz), 125.0, 124.6, 121.3, 118.3, 116.0 (d, J = 22.3 Hz), 112.5, 56.1
1H NMR (300 MHz, DMSO-d6), δ: 7.98–7.88 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 7.74–7.70 (m, 1H, H-5), 7.70–7.64 (m,
AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.49–7.46 (m, 2H, H-7, H-8), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 192.3, 156.6, 144.8, 144.3, 143.9, 129.8, 129.3 (q, J = 32.1 Hz), 126.8 (q, J = 3.8
Hz), 124.7, 124.2 (q, J = 272.3 Hz), 121.3, 118.4, 112.4, 56.1
1H NMR (300 MHz, DMSO-d6), δ: 7.97 (dd, 1H, J = 9.1 Hz, J = 3.0 Hz, H-5), 7.80–7.71 (m, 1H, H-7), 7.60–7.53
(m, 1H, H-8), 7.53–7.47 (m, 2H, H-2ꢀ, H-6ꢀ), 7.47–7.40 (m, 1H, H-4ꢀ), 7.40–7.34 (m, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.5 (d, J = 3.2 Hz), 158.8 (d, J = 241.6 Hz), 146.3 (d, J = 1.7 Hz), 144.6,
140.0, 129.6, 129.0, 128.3, 123.8 (d, J = 24.9 Hz), 122.0 (d, J = 8.9 Hz), 119.4 (d, J = 8.6 Hz), 116.2 (d, J = 26.6
Hz)

E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
359
Table 3. (continued)
Comp.
NMR spectral dataa
IVb
1H NMR (300 MHz, DMSO-d6), δ: 7.96 (dd, 1H, J = 9.2 Hz, J = 3.0 Hz, H-5), 7.78–7.69 (m, 1H, H-7), 7.54 (dd,
1H, J = 9.2 Hz, J = 4.4 Hz, H-8), 7.33–7.27 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.26–7.21 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ),
2.36 (s, 3H, CH3)
13C NMR (75 MHz, DMSO-d6), δ: 191.5 (d, J = 2.9 Hz), 158.7 (d, J = 241.9 Hz), 146.2 (d, J = 1.7 Hz), 144.7,
138.4, 137.5, 130.1, 127.9, 123.8 (d, J = 24.9 Hz), 122.0 (d, J = 8.9 Hz), 119.3 (d, J = 8.3 Hz), 116.2 (d, J = 26.6
Hz), 21.2
IVc
IVd
IVe
IVf
IVg
IVh
1H NMR (300 MHz, DMSO-d6), δ: 7.97 (dd, 1H, J = 9.1 Hz, J = 3.0 Hz, H-5), 7.80–7.71 (m, 1H, H-7), 7.62–7.53
(m, 3H, H-8, H-2ꢀ, H-6ꢀ), 7.46–7.39 (m, 2H, AAꢀ, BBꢀ, H3ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.5 (d, J = 3.1 Hz), 158.8 (d, J = 242.2 Hz), 146.2 (d, J = 1.4 Hz), 144.5,
138.9, 133.6, 130.3, 129.8, 123.9 (d, J = 24.6 Hz), 121.9 (d, J = 8.8 Hz), 119.4 (d, J = 8.3 Hz), 116.2 (d, J = 26.6
Hz)
1H NMR (300 MHz, DMSO-d6), δ: 7.97 (dd, 1H, J = 9.1 Hz, J = 3.0 Hz, H-5), 7.81–7.72 (m, 1H, H-7), 7.62–7.50
(m, 4H, H-8, H-2ꢀ, H-5ꢀ, H-6ꢀ), 7.42–7.36 (m, 1H, H-4ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.4 (d, J = 2.9 Hz), 158.8 (d, J = 242.2 Hz), 146.2 (d, J = 1.7 Hz), 144.5,
141.2, 133.5, 131.3, 129.2, 128.4, 127.4, 124.0 (d, J = 24.7 Hz), 121.9 (d, J = 8.9 Hz), 119.4 (d, J = 8.3 Hz), 116.2
(d, J = 26.6 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 7.98 (dd, 1H, J = 9.1 Hz, J = 3.0 Hz, H-5), 7.82 (d, 1H, J = 8.7 Hz, H-5ꢀ),
7.81–7.73 (m, 2H, H-7, H-2ꢀ), 7.59 (dd, 1H, J = 9.1 Hz, J = 4.4 Hz, H8), 7.44 (dd, 1H, J = 8.7 Hz, J = 2.2 Hz, H-6ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.4 (d, J = 3.1 Hz), 158.8 (d, J = 242.5 Hz), 146.2 (d, J = 1.4 Hz), 144.4,
139.7, 132.0, 131.8, 131.7, 130.6, 129.2, 124.2 (d, J = 24.9 Hz), 121.9 (d, J = 8.9 Hz), 119.5 (d, J = 8.3 Hz), 116.1
(d, J = 26.6 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 7.97 (dd, 1H, J = 9.1 Hz, J = 3.3 Hz, H-5), 7.81–7.68 (m, 3H, H-7, H-2ꢀ, H-6ꢀ),
7.57 (dd, 1H, J = 9.1 Hz, J = 4.4 Hz, H-8), 7.39–7.33 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.4 (d, J = 3.2 Hz), 158.8 (d, J = 241.9 Hz), 146.2 (d, J = 1.7 Hz), 144.5,
139.3, 132.7, 130.6, 124.0 (d, J = 24.6 Hz), 122.2, 121.9 (d, J = 8.8 Hz), 119.4 (d, J = 8.6 Hz), 116.2 (d, J = 26.6
Hz)
1H NMR (300 MHz, DMSO-d6), δ: 7.98 (dd, 1H, J = 9.07 Hz, J = 3.02 Hz, H-5), 7.82–7.73 (m, 1H, H-7), 7.59 (dd,
1H, J = 9.06 Hz, J = 4.39 Hz, H-8), 7.50–740 (m, 2H, H-2ꢀ, H-6ꢀ), 7.40–7.30 (m, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.9, 162.0 (d, J = 245.1 Hz), 158.8 (d, J = 242.2 Hz), 146.3 (d, J = 3.2 Hz),
144.7 (d, J = 2.0 Hz), 136.3 (d, J = 3.2 Hz), 130.6 (d, J = 9.2 Hz), 123.9 (d, J = 24.6 Hz), 122.0 (d, J = 8.9 Hz),
119.4 (d, J = 8.6 Hz), 116.6 (d, J = 23.2 Hz), 116.2 (d, J = 26.3 Hz)
1H NMR (300 MHz, CDCl3), δ: 8.00 (dd, 1H, J = 8.52 Hz, J = 3.02 Hz, H-5), 7.58–7.40 (m, 2H, H-7, H-6ꢀ), 7.31
(dd, 1H, J = 9.07 Hz, J = 4.12 Hz, H-8), 7.22 (ddd, 1H, J = 8.52 Hz, J = 2.47 Hz, J = 0.82 Hz, H-2ꢀ), 7.09–7.04
(m, 1H, H-5ꢀ), 7.01 (dt, 1H, J = 8.51 Hz, J = 2.20 Hz, H-4ꢀ)
13C NMR (75 MHz, CDCl3), δ: 190.1, 163.3 (d, J = 248.9 Hz), 159.7 (d, J = 246.8 Hz), 145.5 (d, J = 2.0 Hz),
144.6, 140.0 (d, J = 10.0 Hz), 131.1 (d, J = 8.8 Hz), 123.8 (d, J = 1.7 Hz), 123.7 (d, J = 23.5 Hz), 121.7 (d, J =
8.9 Hz), 118.7 (d, J = 8.3 Hz), 117.4 (d, J = 26.6 Hz), 116.7 (d, J = 20.9 Hz), 115.9 (d, J = 23.7 Hz)
IVi
1H NMR (300 MHz, DMSO-d6), δ: 7.99 (dd, 1H, J = 9.07 Hz, J = 3.30 Hz, H-5), 7.96–7.90 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ,
H-5ꢀ), 7.84–7.75 (m, 1H, H-7), 7.70–7.64 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.62 (dd, 1H, J = 9.06 Hz, J = 4.39 Hz,
H-8)
13C NMR (75 MHz, DMSO-d6), δ: 191.6 (d, J = 3.1 Hz), 158.9 (d, J = 242.5 Hz), 146.4 (d, J = 1.7 Hz), 144.5,
143.7, 129.7, 129.4 (q, J = 32.1 Hz), 126.9 (d, J = 4.0 Hz), 124.2 (q, J = 272.6 Hz), 124.1 (d, J = 24.9 Hz), 122.0
(d, J = 8.9 Hz), 119.5 (d, J = 8.6 Hz), 116.1 (d, J = 26.6 Hz)
Vg
Vh
Vi
1H NMR (300 MHz, DMSO-d6), δ: 8.24–8.21 (m, 1H, H-5), 7.97–7.91 (m, 1H, H-7), 7.64–7.52 (m, 1H, H-8), 7.38–7.23
(m, 4H, H-2ꢀ, H-3ꢀ, H-5ꢀ, H-6ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.6, 162.0 (d, J = 245.1 Hz), 148.7, 144.5, 136.2 (d, J = 3.1 Hz), 136.0, 130.6
(d, J = 8.9 Hz), 130.0, 129.8, 122.2, 119.3, 116.6 (d, J = 23.2 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 8.24 (m, 1H, H5), 7.96–7.91 (m, 1H, H-7), 7.65–7.53 (m, 2H, H-2ꢀ, H-6ꢀ), 7.39–7.23
(m, 3H, H-8, H-4ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 191.2, 162 (d, J = 244.2 Hz), 148.6, 144.3, 141.2 (d, J = 10.9 Hz), 136.1, 131.3
(d, J = 8.8 Hz), 129.9, 124.8 (d, J = 3.2 Hz), 122.1, 119.3, 116.2 (d, J = 20.7 Hz), 115.8 (d, J = 23.8 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 8.24–8.22 (m, 1H, H-5), 7.97–7.91 (m, 3H, H-7, H-3ꢀ, H-5ꢀ), 7.70–7.62 (m, 2H,
H-2ꢀ, H-6ꢀ), 7.59 (d, 1H, J = 8.79 Hz, H-8)
13C NMR (75 MHz, DMSO-d6), δ: 191.3, 148.7, 144.3, 143.5, 136.1, 129.9, 129.8, 129.7, 129.5 (q, J = 32.1 Hz),
126.9 (d, J = 3.8 Hz), 124.2 (q, J = 272.3 Hz), 122.1, 119.3
VIa
VIb
1H NMR (300 MHz, CDCl3), δ: 8.45 (d, 1H, J = 2.51 Hz, H-5), 7.81 (dd, 1H, J = 8.82 Hz, J = 2.51 Hz, H-7),
7.61–7.48 (m, 3H, H-3ꢀ, H-4ꢀ, H-5ꢀ), 7.29–7.18 (m, 3H, H-8, H-2ꢀ, H-6ꢀ)
13C NMR (75 MHz, CDCl3), δ: 185.4, 176.9, 148.5, 143.0, 138.6, 134.2, 130.0, 129.1, 127.6, 123.3, 119.5, 118.0
1H NMR (300 MHz, CDCl3), δ: 8.45 (d, 1H, J = 2.40 Hz, H-5), 7.81 (dd, 1H, J = 8.77 Hz, J = 2.40 Hz, H-7),
7.39–7.34 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.25 (d, 1H, J = 8.77 Hz, H-8), 7.12–7.06 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ),
2.46 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 185.5, 177.0, 148.5, 140.5, 139.2, 138.6, 134.2, 130.7, 127.2, 123.3, 119.5, 118.0, 21.6

360
E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Table 3. (continued)
Comp.
NMR spectral dataa
VIc
1H NMR (300 MHz, CDCl3), δ: 8.44 (d, 1H, J = 2.43 Hz, H-5), 7.82 (dd, 1H, J = 8.77 Hz, J = 2.43 Hz, H-7),
7.55–7.49 (m, 2H, H-2ꢀ, H-6ꢀ), 7.26 (d, 1H, J = 8.77 Hz, H-8), 7.17–7.11 (m, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, CDCl3), δ: 185.3, 176.7, 148.5, 141.3, 138.8, 135.1, 134.2, 130.4, 129.1, 123.2, 119.6, 118.1
VId
1H NMR (300 MHz, CDCl3), δ: 8.43 (d, 1H, J = 2.43 Hz, H-5), 7.82 (dd, 1H, J = 8.80 Hz, J = 2.43 Hz, H-7),
7.51–7.45 (m, 2H, H-2ꢀ, H-4ꢀ), 7.25 (d, 1H, J = 8.80 Hz, H-8), 7.23–7.21 (m, 1H, H-6ꢀ), 7.14–7.08 (m, 1H, H-5ꢀ)
13C NMR (75 MHz, CDCl3), δ: 185.1, 176.5, 148.4, 143.6, 138.8, 135.4, 134.1, 130.9, 129.4, 128.2, 126.1, 123.1,
119.6, 118.1
VIe
1H NMR (300 MHz, CDCl3), δ: 8.43 (d, 1H, J = 2.46 Hz, H-5), 7.83 (dd, 1H, J = 8.76 Hz, J = 2.46 Hz, H-7), 7.62
(d, 1H, J = 8.70 Hz, H-5ꢀ), 7.32 (d, 1H, J = 2.45 Hz, H-2ꢀ), 7.25 (d, 1H, J = 8.76 Hz, H-8), 7.06 (dd, 1H, J = 8.70
Hz, J = 2.45 Hz, H-6ꢀ)
13C NMR (75 MHz, CDCl3), δ: 185.1, 176.4, 148.4, 141.6, 138.9, 134.1, 134.0, 133.6, 131.7, 130.0, 127.4, 123.0,
119.7, 118.1
VIf
1H NMR (300 MHz, CDCl3), δ: 8.44 (d, 1H, J = 2.38 Hz, H-5), 7.82 (dd, 1H, J = 8.80 Hz, J = 2.38 Hz, H-7),
7.70–7.66 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.25 (d, 1H, J = 8.80 Hz, H-8), 7.10–7.06 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, CDCl3), δ: 185.3, 176.6, 148.5, 141.8, 138.8, 134.2, 133.4, 130.0, 129.5, 123.2, 119.6, 118.1
VIg
1H NMR (300 MHz, CDCl3), δ: 8.44 (d, 1H, J = 2.47 Hz, H-5), 7.82 (dd, 1H, J = 8.79 Hz, J = 2.47 Hz, H-7),
7.29–7.13 (m, 5H, H-8, H-2ꢀ H-3ꢀ, H-5ꢀ, H-6ꢀ)
13C NMR (75 MHz, CDCl3), δ: 185.7, 177.1, 162.5 (d, J = 249.9 Hz), 148.6, 138.9, 138.9, 134.3, 129.7 (d, J = 8.9
Hz), 123.3, 119.7, 118.1, 117.3 (d, J = 23.5 Hz)
VIh
VIi
1H NMR (300 MHz, CDCl3), δ: 8.43 (d, 1H, J = 2.47 Hz, H-5), 7.82 (dd, 1H, J = 8.79 Hz, J = 2.47 Hz, H-7),
7.57–7.48 (m, 1H, H-6ꢀ), 7.26 (d, overlapped, 1H, J = 8.79 Hz, H-8), 7.25–7.17 (m, 1H, H-2ꢀ), 7.04–6.99 (m, 1H,
H-5ꢀ), 6.96 (dt, 1H, J = 8.52 Hz, J = 2.20 Hz, H-4ꢀ)
13C NMR (75 MHz, CDCl3), δ: 185.4, 176.7, 163.5 (d, J = 248.8 Hz), 148.6, 144.1 (d, J = 10.3 Hz), 138.9, 134.2,
131.3 (d, J = 8.9 Hz), 123.8 (d, J = 3.4 Hz), 123.3, 119.7, 118.2, 116.5 (d, J = 20.9 Hz), 115.8 (d, J = 24.1 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 8.28 (d, 1H, J = 2.47 Hz, H-5), 8.09 (dd, 1H, J = 8.79 Hz, J = 2.48 Hz, H-7),
7.96–7.89 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 7.65–7.59 (m, AAꢀ, BBꢀ overlapped, 2H, H-2ꢀ, H-6ꢀ), 7.60 (d, overlapped,
1H, J = 8.79 Hz, H-8)
13C NMR (75 MHz, DMSO-d6), δ: 186.9, 177.6, 149.2, 147.1, 139.2, 132.2, 129.5, 129.3 (q, J = 32.1 Hz), 127.2 (q,
J = 3.7 Hz), 124.2 (q, J = 278.6 Hz), 124.0, 119.1, 118.7
VIIa
VIIb
1H NMR (300 MHz, CDCl3), δ: 8.16–8.13 (m, 1H, H-5), 7.62–7.46 (m, 4H, H-7, H-3ꢀ, H-4ꢀ, H-5ꢀ), 7.29–7.19 (m, 3H,
H-8, H-2ꢀ, H-6ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 187.6, 178.0, 148.0, 143.4, 137.2, 136.8, 131.6, 129.9, 129.0, 127.9, 121.9, 116.0, 20.9
1H NMR (300 MHz, CDCl3), δ: 8.15–8.13 (m, 1H, H-5), 7.57–7.52 (m, 1H, H-7), 7.40–7.32 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ,
H-6ꢀ), 7.26 (d, 1H, J = 8.52 Hz, H-8), 7.14–7.07 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 2.46 (s, 3H, CH3), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 187.7, 178.1, 148.0, 140.9, 139.1, 137.2, 136.7, 131.7, 130.7, 127.5, 121.9, 116.0, 21.4,
20.9
VIIc
VIId
VIIe
1H NMR (300 MHz, CDCl3), δ: 8.14–8.11 (m, 1H, H-5), 7.55 (dd, 1H, J = 8.52 Hz, J = 2.20 Hz, H-7), 7.54–7.49
(m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.26 (d, 1H, J = 7.51 Hz, H-8), 7.18–7.12 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 2.45 (s,
3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 187.5, 177.8, 148.0, 141.7, 137.4, 136.9, 134.9, 131.6, 130.3, 129.4, 121.7, 116.0, 20.9
1H NMR (300 MHz, CDCl3), δ: 8.14–8.11 (m, 1H, H-5), 7.58–7.53 (m, 1H, H-7), 7.51–7.43 (m, 2H, H-2ꢀ, H-6ꢀ), 7.26
(d, 1H, J = 8.51 Hz, H-8), 7.25–7.22 (m, 1H, H-5ꢀ), 7.15–7.10 (m, 1H, H-4ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 187.3, 177.6, 148.0, 144.0, 137.4, 136.9, 135.4, 131.6, 130.9, 129.3, 128.5, 126.4,
121.7, 116.1, 20.9
1H NMR (300 MHz, CDCl3), δ: 8.12–8.10 (m, 1H, H-5), 7.61 (d, 1H, J = 8.52 Hz, H-5ꢀ), 7.59–7.54 (m, 1H, H-7),
7.33 (d, 1H, J = 2.47 Hz, H-2ꢀ), 7.26 (d, 1H, J = 8.24 Hz, H-8), 7.08 (dd, 1H, J = 8.52 Hz, J = 2.47 Hz, H-6ꢀ), 2.45
(s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 187.3, 177.5, 148.0, 142.1, 137.6, 137.1, 134.0, 133.5, 131.7, 131.6, 130.3, 127.7,
121.6, 116.1, 21.0
VIIf
VIIg
VIIh
1H NMR (300 MHz, CDCl3), δ: 8.14–8.11 (m, 1H, H-5), 7.71–7.64 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.55 (ddd, 1H,
J = 8.38 Hz, J = 2.19 Hz, J = 0.55 Hz, H-7), 7.26 (d, 1H, J = 8.38 Hz, H-8), 7.12–7.06 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ,
H-5ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 187.4, 177.7, 148.0, 142.2, 137.4, 136.9, 133.3, 131.6, 129.7, 123.1, 121.8, 116.1, 21.0
1H NMR (300 MHz, (CDCl3), δ: 8.14–8.11 (m, 1H, H5), 7.55 (ddd, 1H, J = 8.52 Hz, J = 2.20 Hz, J = 0.50 Hz,
H-7), 7.27–7.15 (m, 4H, H-2ꢀ H-3ꢀ, H-5ꢀ, H-6ꢀ), 7.25 (d, overlapped, 1H, J = 8.52 Hz, H-8), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3), δ: 187.7, 178.0, 162.4 (d, J = 249.6 Hz), 148.0, 139.2 (d, J = 3.5 Hz), 137.3, 136.9,
131.7, 129.8 (d, J = 8.6 Hz), 121.8, 117.1 (d, J = 23.2 Hz), 116.0, 20.9
1H NMR (300 MHz, DMSO-d6), δ: 8.14–8.11 (m, 1H, H-5), 7.58–7.47 (m, 2H, H-2ꢀ, H-6ꢀ), 7.25 (d, 1H, J = 8.24 Hz,
H-8), 7.20 (tdd, 1H, J = 8.24 Hz, J = 2.47 Hz, J = 0.82 Hz, H-5ꢀ), 7.05–7.00 (m, 1H, H-7), 6.97 (dt, 1H, J = 8.79
Hz, J = 2.20 Hz, H-4ꢀ), 2.45 (s, 3H, CH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.3, 177.5, 163.5 (d, J = 248.6 Hz), 148.0, 144.2 (d, J = 10.3 Hz), 137.4, 136.9,
131.5, 131.1 (d, J = 8.9 Hz), 123.9 (d, J = 3.1 Hz), 121.7, 116.2 (d, J = 26.0 Hz), 116.1 (d, J = 23.8 Hz), 20.9

E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
361
Table 3. (continued)
Comp.
NMR spectral dataa
VIIi
1H NMR (300 MHz, DMSO-d6), δ: 8.05–8.02 (m, 1H, H-5), 7.95–7.87 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 7.74 (dd, 1H,
J = 8.38 Hz, J = 2.20 Hz, H-7), 7.66–7.59 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.49 (d, 1H, J = 8.38 Hz, H-8), 2.42 (s,
3H, CH3)
13C NMR (75 MHz, DMSO-d6), δ: 188.3, 178.1, 148.3, 147.3, 137.9, 136.7, 130.7, 129.7, 129.1 (q, J = 32.1 Hz),
127.1 (q, J = 3.8 Hz), 124.2 (q, J = 272.3 Hz), 122.1, 116.3, 20.6
VIIIa
VIIIb
VIIIc
VIIId
VIIIe
1H NMR (300 MHz, DMSO-d6), δ: 7.63 (d, 1H, J = 2.74 Hz, H-5), 7.57–7.46 (m, 4H, H-7, H-8, H-2ꢀ, H-6ꢀ), 7.45–7.30
(m, 3H, H-3ꢀ, H-4ꢀ, H-5ꢀ), 3.84 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.9, 178.0, 157.3, 144.8, 144.1, 129.8, 128.8, 128.2, 124.9, 122.9, 118.2, 112.2,
56.2
1H NMR (300 MHz, DMSO-d6), δ: 7.61 (d, 1H, J = 2.75 Hz, H-5), 7.56–7.44 (m, 2H, H-7, H-8), 7.33–7.25 (m, AAꢀ,
BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.23–7.15 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 3.84 (s, 3H, OCH3), 2.36 (s, 3H, CH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.9, 178.0, 157.3, 144.7, 141.6, 138.1, 130.4, 127.9, 124.9, 122.9, 118.1, 112.3,
56.1, 21.1
1H NMR (300 MHz, DMSO-d6), δ: 7.64–7.47 (m, 5H, H-5, H-7, H-8, H-2ꢀ, H-6ꢀ), 7.42–7.36 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ,
H-5ꢀ), 3.85 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 188.0, 177.9, 157.3, 144.8, 142.9, 133.3, 130.4, 130.0, 125.0, 122.9, 118.2, 112.1,
56.2
1H NMR (300 MHz, DMSO-d6), δ: 7.62 (d, 1H, J = 2.75 Hz, H5), 7.59–7.46 (m, 5H, H-7, H-8, H-2ꢀ, H-5ꢀ, H-6ꢀ),
7.39–7.33 (m, 1H, H-4ꢀ), 3.85 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.9, 177.9, 157.3, 145.1, 144.7, 133.7, 131.5, 128.9, 128.5, 127.4, 125.0, 122.9,
118.2, 112.1, 56.2
1H NMR (300 MHz, DMSO-d6), δ: 7.82 (d, 1H, J = 8.79 Hz, H-5ꢀ), 7.77 (d, 1H, J = 2.19 Hz, H-5), 7.62 (d, 1H, J
= 3.03 Hz, H-2ꢀ), 7.58 (d, 1H, J = 8.52 Hz, H-8), 7.52 (dd, 1H, J = 8.79 Hz, J = 3.02 Hz, H-6ꢀ), 7.43 (dd, 1H, J =
8.52 Hz, J = 2.20 Hz, H-7), 3.85 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.9, 177.8, 157.4, 144.7, 143.6, 132.0, 131.9, 131.6, 130.7, 129.2, 125.1, 122.9,
118.2, 112.1, 56.2
VIIIf
1H NMR (300 MHz, DMSO-d6), δ: 7.75–7.69 (m, AAꢀ, BB, 2H, H-2ꢀ, H-6ꢀ), 7.62 (d, 1H, J = 2.75 Hz, H-5), 7.56
(dd, 1H, J = 9.07 Hz, J = 0.55 Hz, H-8), 7.50 (dd, 1H, J = 9.06 Hz, J = 2.75 Hz, H-7), 7.36–7.30 (m, AAꢀ, BBꢀ,
2H, H-3ꢀ, H-5ꢀ), 3.85 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.9, 177.9, 157.3, 144.8, 143.4, 133.0, 130.7, 125.0, 122.9, 121.9, 118.2, 112.1,
56.2
VIIIg
VIIIh
VIIIi
1H NMR (300 MHz, DMSO-d6), δ: 7.62 (d, 1H, J = 3.02 Hz, H5), 7.58–7.47 (m, 2H, H-2ꢀ, H-6ꢀ), 7.44–7.32 (m, 4H,
H-7, H-8, H-3ꢀ, H-5ꢀ), 3.85 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 188.1, 178.1, 161.9 (d, J = 245.3 Hz), 157.3, 144.7, 140.4 (d, J = 3.1 Hz), 130.5
(d, J = 8.9 Hz), 125.0, 122.9, 118.2, 116.8 (d, J = 23.2 Hz), 112.2, 56.2
1H NMR (300 MHz, DMSO-d6), δ: 7.63 (d, 1H, J = 3.03 Hz, H-5), 7.61–7.54 (m, 2H, H-2ꢀ, H-6ꢀ), 7.51 (dd, overlapped,
1H, J = 9.07 Hz, J = 3.03 Hz, H7), 7.36–7.20 (m, 3H, H-8, H-4ꢀ, H-5ꢀ), 3.85 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.8, 177.8, 162.9 (d, J = 243.9 Hz), 157.3, 145.2 (d, J = 11.2 Hz), 144.7, 131.5
(d, J = 8.8 Hz), 124.9 (d, J = 16.1 Hz), 124.9, 122.9, 118.2, 116.0 (d, J = 22.6 Hz), 112.1, 56.2
1H NMR (300 MHz, DMSO-d6), δ: 7.95–7.88 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ), 7.67–7.60 (m, AAꢀ, BBꢀ overlapped,
2H, H-2ꢀ, H-6ꢀ), 7.61 (d, overlapped, 1H, J = 9.48 Hz, H-8), 7.56 (bs, 1H, H-5), 7.52 (dd, 1H, J = 9.48 Hz, J = 3.02
Hz, H-7), 3.85 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO-d6), δ: 188.0, 177.8, 157.4, 147.4, 144.8, 129.7, 129.1 (q, J = 31.8 Hz), 127.1 (q, J = 3.8
Hz), 125.1, 124.2 (q, J = 272.3 Hz), 123.0, 118.2, 112.0, 56.2
IXa
IXb
1H NMR (300 MHz, DMSO-d6), δ: 7.97 (dd, 1H, J = 8.8 Hz, J = 3.0 Hz, H-5), 7.83–7.74 (m, 1H, H-7), 7.68–7.62
(m, 1H, H-8), 7.55–7.46 (m, 2H, H-2ꢀ, H-6ꢀ), 7.45–7.37 (m, 1H, H-4ꢀ), 7.36–7.29 (m, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.1 (d, J = 3.1 Hz), 177.8, 159.3 (d, J = 243.4 Hz), 146.5 (d, J = 1.4 Hz),
143.8, 129.9, 128.9, 128.1, 124.2 (d, J = 24.9 Hz), 123.6 (d, J = 9.1 Hz), 119.2 (d, J = 8.5 Hz), 116.2 (d, J = 26.6
Hz)
1H NMR (300 MHz, DMSO-d6), δ: 7.87 (dd, 1H, J = 8.9 Hz, J = 3.0 Hz, H-5), 7.81–7.72 (m, 1H, H-7), 7.63 (dd,
1H, J = 8.9 Hz, J = 4.4 Hz, H-8), 7.33–7.26 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.21–7.15 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ),
2.35 (s, 3H, CH3)
13C NMR (75 MHz, DMSO-d6), δ: 187.0 (d, J = 3.2 Hz), 177.8, 159.3 (d, J = 243.4 Hz), 146.4 (d, J = 1.4 Hz),
141.3, 138.3, 130.4, 127.7, 124.2 (d, J = 24.9 Hz), 123.5 (d, J = 9.2 Hz), 119.2 (d, J = 8.3 Hz), 116.3 (d, J = 26.6
Hz), 21.2
IXc
1H NMR (300 MHz, DMSO-d6), δ: 7.89 (dd, 1H, J = 8.9 Hz, J = 3.0 Hz, H-5), 7.83–7.75 (m, 1H, H-7), 7.66 (dd,
1H, J = 8.9 Hz, J = 4.4 Hz, H-8), 7.62–7.54 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.42–7.33 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.1 (d, J = 3.2 Hz), 177.7, 159.3 (d, J = 243.6 Hz), 146.5 (d, J = 1.4 Hz),
142.6, 133.4, 130.2, 130.1, 124.3 (d, J = 24.9 Hz), 123.5 (d, J = 9.1 Hz), 119.2 (d, J = 8.6 Hz), 116.2 (d, J = 26.6
Hz)

362
E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Table 3. (continued)
Comp.
NMR spectral dataa
IXd
IXe
IXf
1H NMR (300 MHz, DMSO-d6), δ: 7.90 (dd, 1H, J = 9.0 Hz, J = 3.0 Hz, H-5), 7.84–7.75 (m, 1H, H-7), 7.67 (dd,
1H, J = 9.0 Hz, J = 4.4 Hz, H-8), 7.60–7.46 (m, 3H, H-2ꢀ, H-5ꢀ, H-6ꢀ), 7.38–7.32 (1H, m, H-4ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.1 (d, J = 3.2 Hz), 177.7, 159.4 (d, J = 243.6 Hz), 146.5 (d, J = 1.7 Hz),
144.8, 133.8, 131.6, 129.1, 128.3, 127.3, 124.4 (d, J = 25.2 Hz), 123.5 (d, J = 9.2 Hz), 119.3 (d, J = 8.6 Hz), 116.2
(d, J = 26.6 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 7.91 (dd, 1H, J = 9.1 Hz, J = 2.8 Hz, H-5), 7.86–7.77 (m, 2H, H-7, H-5ꢀ), 7.74
(d, 1H, J = 2.3 Hz, H-2ꢀ), 7.68 (dd, 1H, J = 9.1 Hz, J = 4.4 Hz, H-8), 7.41 (dd, 1H, J = 8.5 Hz, J = 2.3 Hz, H-6ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.1 (d, J = 3.2 Hz), 177.7, 159.4 (d, J = 244.0 Hz), 146.4 (d, J = 1.4 Hz),
143.3, 132.1, 132.0, 131.8, 130.5, 129.1, 124.5 (d, J = 24.9 Hz), 123.5 (d, J = 9.1 Hz), 119.3 (d, J = 8.3 Hz), 116.2
(d, J = 26.6 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 7.89 (dd, 1H, J = 8.9 Hz, J = 3.3 Hz, H-5), 7.83–7.75 (m, 1H, H-7), 7.75–7.69
(m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ), 7.66 (dd, 1H, J = 8.9 Hz, J = 4.1 Hz, H-8), 7.36–7.27 (m, AAꢀ, BBꢀ, 2H, H-3ꢀ, H-5ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.0 (d, J = 3.1 Hz), 177.6, 159.3 (d, J = 243.7 Hz), 146.5 (d, J = 1.4 Hz),
143.1, 133.0, 130.5, 124.3 (d, J = 24.9 Hz), 123.5 (d, J = 9.1 Hz), 122.0, 119.2 (d, J = 8.3 Hz), 116.2 (d, J = 26.6
Hz)
IXg
IXh
1H NMR (300 MHz, CDCl3), δ: 8.00 (dd, 1H, J = 8.52 Hz, J = 3.02 Hz, H-5), 7.50–7.42 (m, 1H, H-7), 7.37 (dd,
1H, J = 9.07 Hz, J = 4.40 Hz, H-8), 7.28–7.14 (m, 4H, H-2ꢀ, H-3ꢀ, H-5ꢀ, H-6ꢀ)
13C NMR (75 MHz, CDCl3), δ: 186.2 (d, J = 3.2 Hz), 177.3, 162.5 (d, J = 249.9 Hz), 160.0 (d, J = 247.9 Hz),
146.0, 139.0 (d, J = 3.4 Hz), 129.7 (d, J = 8.8 Hz), 123.8 (d, J = 24.9 Hz), 123.2 (d, J = 8.8 Hz), 118.5 (d, J = 8.3
Hz), 117.4 (d, J = 26.6 Hz), 117.3 (d, J = 23.2 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 8.00 (dd, 1H, J = 8.52 Hz, J = 3.02 Hz, H-5), 7.57–7.42 (m, 2H, H-2ꢀ, H-6ꢀ),
7.37 (dd, 1H, J = 9.06 Hz, J = 4.40 Hz, H-8), 7.21 (tdd, 1H, J = 8.24 Hz, J = 2.47 Hz, J = 0.83 Hz, H-5ꢀ), 7.04–6.99
(m, 1H, H-7), 6.96 (dt, 1H, J = 8.24 Hz, J = 2.47 Hz, H-4ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 185.8, 176.9, 163.6 (d, J = 248.9 Hz), 160.0 (d, J = 247.7 Hz), 146.0, 144.0 (d,
J = 10.3 Hz), 131.3 (d, J = 8.9 Hz), 123.9 (d, J = 20.1, Hz), 123.7 (d, J = 7.8 Hz), 123.2 (d, J = 9.1 Hz), 118.5 (d,
J = 8.3 Hz), 117.3 (d, J = 26.6 Hz), 116.5 (d, J = 20.9 Hz), 115.9 (d, J = 24.1 Hz)
IXi
1H NMR (300 MHz, DMSO-d6), δ: 7.96–7.89 (m, AAꢀ, BBꢀ overlapped, 3H, H-5, H-3ꢀ, H-5ꢀ), 7.88–7.79 (m, 1H,
H-7), 7.72 (dd, 1H, J = 9.07 Hz, J = 4.40 Hz, H-8), 7.66–7.60 (m, AAꢀ, BBꢀ, 2H, H-2ꢀ, H-6ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.3 (d, J = 3.2 Hz), 177.8, 159.4 (d, J = 244.2 Hz), 147.3, 146.6 (d, J = 1.7
Hz), 129.6, 129.2 (q, J = 31.8 Hz), 127.2 (d, J = 3.7 Hz), 124.4 (d, J = 25.2 Hz), 124.2 (q, J = 270.9 Hz), 123.6 (d,
J = 9.2 Hz), 119.3 (d, J = 8.6 Hz), 116.1 (d, J = 26.6 Hz)
Xg
Xh
Xi
1H NMR (300 MHz, DMSO-d6), δ: 8.10 (d, 1H, J = 2.47 Hz), 7.92 (dd, 1H, J = 8.79 Hz, J = 2.47 Hz, H-7), 7.62
(d, 1H, J = 8.79 Hz), 7.40–7.27 (m, 4H, H-2ꢀ, H-3ꢀ, H-5ꢀ, H-6ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.2, 178.0, 161.9 (d, J = 245.1 Hz), 148.8, 140.1 (d, J = 3.5 Hz), 136.3, 130.8,
130.5 (d, J = 8.9 Hz), 130.0, 123.6, 119.0, 116.9 (d, J = 23.2 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 8.14 (d, 1H, J = 2.75 Hz, H-5), 7.97 (dd, 1H, J = 8.79 Hz, J = 2.74 Hz, H-7),
7.66 (d, 1H, J = 8.79 Hz, H-8), 7.62–7.52 (m, 1H, H-6ꢀ), 7.34–7.25 (m, 2H, H-2ꢀ, H-5ꢀ), 7.25–7.20 (m, 1H, H-4ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 186.8, 177.6, 162.9 (d, J = 244.0 Hz), 148.7, 144.8 (d, J = 10.9 Hz), 136.4, 131.6
(d, J = 8.8 Hz), 130.8, 129.9, 124.7 (d, J = 2.9 Hz), 123.5, 119.0, 116.1 (d, J = 20.7 Hz), 115.7 (d, J = 24.1 Hz)
1H NMR (300 MHz, DMSO-d6), δ: 8.16–8.14 (m, 1H, H-5), 8.01–7.89 (m, 3H, H-7, H-3ꢀ, H-5ꢀ), 7.68 (d, 1H, J =
8.79 Hz, H-8), 7.65–7.59 (m, 2H, H-2ꢀ, H-6ꢀ)
13C NMR (75 MHz, DMSO-d6), δ: 187.0, 177.6, 148.8, 147.2, 136.4, 130.9, 129.8, 129.5, 129.2 (q, J = 32.1 Hz),
127.2 (q, J = 3.7 Hz), 124.2 (q, J = 272.3 Hz), 123.6, 119.0
a) Atom numbering corresponds to the numbering in Fig. 1.
and 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-dithiones
were synthesised by treating 3-phenyl-2H-1,3-benz-
oxazine-2,4(3H)-diones with phosphorous pentasul-
phide (Fig. 1). The published methods include thion-
ation using P2S5 and Lawesson’s reagent (Wagner
et al., 1966; Saeed & Ashraf, 2008). The start-
ing 3-phenyl-2H-1,3-benzoxazine-2(3H )-diones are de-
scribed in the literature (Waisser et al., 2001a, 2001b).
An overview, characterisation, and spectral data of the
compounds as prepared are summarised in Tables 1–
3. The structural evidence and the physical properties
of compounds Va–Vf and Xa–Xf can be found in the
literature (Waisser et al., 2000).
compounds was investigated against M. tuberculo-
sis CNCTC My 331/88 (identical with H37RV and
ATCC 27294), M. kansasii CNCTC My 235/80 (iden-
tical with ATCC 12 478, resistant to INH), M. avium
CNCTC My 330/88 (identical with ATCC 25291, re-
sistant to INH) and M. kansasii 6509/96. All the
strains were obtained from the Czech National Col-
lection of Type Cultures (CNCTC), National Institute
of Public Health, Prague. The only exception was M.
kansasii 6509/96, which was clinically isolated. The
dilution micromethod was used for determination of
the minimum inhibitory concentrations (MIC) and the
MIC values for standard INH were included for com-
parison. An overview of the biological activity along
The in-vitro antimycobacterial activity of the

E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
363
Table 4. In-vitro antimycobacterial activity of 3-phenyl-4-
thioxo-2H-1,3-benzoxazine-2(3H )-ones
Table 5. In-vitro antimycobacterial activity of 3-phenyl-2H-
1,3-benzoxazine-2,4(3H )-dithiones
a
a
MIC/(µmol L−1
)
MIC/(µmol L−1
)
Comp.
M. tuberculosis M. avium
My 331/88 My 330/88
Comp.
M. kansasii M. kansasii
M. tuberculosis M. avium
M. kansasii M. kansasii
My 235/80 6509/96
My 235/80
6509/96
My 331/88
My 330/88
Ia
Ib
Ic
Id
Ie
If
Ig
Ih
Ii
IIa
IIb
IIc
IId
IIe
IIf
IIg
IIh
IIi
IIIa
IIIb
IIIc
IIId
IIIe
IIIf
IIIg
IIIh
IIIi
IVa
IVb
IVc
IVd
IVe
IVf
IVg
IVh
IVj
Vad
Vbd
Vcd
Vdd
Ved
Vfb
Vg
0.5/1
0.5/1
0.5/1
0.5/1
1/4
1/2
0.5/2
2/2
8/16
32/ndb
16/32
16/32
32/62.5
32/32
ndb
16/32
32/62.5
4/4
1/1
2/2
4/4
8/16
4/4
4/4
8/16
32/32
32/32
32/32
32/ndb
62.5/62.5
62.5/ndb
62.5/62.5
32/62.5
62,5/125
125/125
16/ntc
8/ntc
4/8
4/8
8/8
4/8
8/16
16/16
32/32
8/32
16/32
16/16
16/32
8/16
2/4
2/4
4/8
2/4
4/8
4/4
8/16
8/8
8/16
2/4
2/4
2/2
2/4
4/4
2/4
4/8
4/4
8/16
8/8
8/16
2/4
2/2
VIa
VIb
VIc
VId
VIe
VIf
VIg
VIh
VIi
VIIa
VIIb
VIIc
VIId
VIIe
VIIf
VIIg
VIIh
VIIi
VIIIa
VIIIb
VIIIc
VIIId
VIIIe
VIIIf
VIIIg
VIIIh
VIIIi
IXa
IXb
2/2
0.5/1
1/2
1/1
2/2
1/2
1/2
2/4
4/4
0.5/1
0.5/1
0.5/1
0.5/0.5
0.5/0.5
0,5/0,5
2/8
4/8
4/8
ndb
8/16
8/16
16/32
16/32
8/16
16/32
62.5/ndb
16/32
4/4
1/2
2/2
2/4
1/2
1/2
1/2
16/16
2/4
1/1
8/16
16/32
32/32
16/32
32/62.5
32/32
16/16
16/32
62.5/125
4/4
2/2
4/4
2/4
8/16
4/4
4/4
8/8
32/32
ndb
16/32
62.5/62.5
ndb
62.5/ndb
62.5/62.5
32/ndb
ndb
62.5/125
8/ntc
16/ntc
16/ntc
8/ntc
16/ntc
16/ntc
16/16
16/16
32/32
8/16
16/ntc
16/16
1/2
2/2
4/4
4/4
2/4
4/8
2/4
4/4
4/4
2/4
2/4
4/4
8/16
8/8
32/32
2/4
2/2
4/4
8/16
16/16
16/ndb
2/4
2/4
4/4
2/2
8/8
4/4
8/16
16/16
16/16
ndb
16/16
32/32
16/32
32/32
32/32
16/16
ndb
32/32
4/ntc
8/8
2/4
4/ntc
4/4
2/4
1/2
0.5/1
0.5/0.5
0.5/0.5
0.5/0.5
0.5/0.5
4/8
2/4
2/4
4/8
2/4
2/4
2/4
8/8
4/4
4/8
2/4
8/8
4/4
8/16
16/32
16/32
16/32
16/16
16/16
16/32
62.5/62.5
32/32
16/32
32/32
32/62.5
0.5/ntc
1/ntc
2/ntc
1/ntc
2/ntc
2/ntc
8/16
16/16
8/8
8/8
4/8
4/8
4/8
16/16
16/32
8/16
8/8
16/16
16/32
62.5/62.5
32/32
16/16
32/32
32/62.5
0.5/1
1/2
1/2
0.5/1
2/2
1/4
8/8
16/16
8/8
0.5/1
1/1
1/1
1/1
8/16
16/16
ndb
8/16
16/32
8/8
32/32
32/32
16/16
ndb
32/32
1/2
2/4
4/4
2/2
1/2
2/4
8/8
16/16
8/8
1/1
0.5/1
1/2
2/4
1/1
1/2
8/8
8/8
16/16
4/4
16/32
16/32
8/16
16/32
16/32
32/32
16/16
16/32
32/32
2/4
1/2
1/1
2/2
0.5/1
1/2
IXc
IXd
IXe
IXf
IXg
4/ntc
8/16
16/16
8/8
2/4
2/4
2/8
16/32
2/4
1/2
8/16
2/4
IXh
IXj
1/1
2/4
4/8
2/4
2/4
2/4
2/4
Xad
Xbd
Xcd
Xdd
Xed
Xfd
0.5/1
0.5/0.5
0.5/1
0.5/1
0.5/1
2/4
2/4
4/4
0.5/1
0.5/0.5
0.5/1
0.5/1
1/2
0.5/0.5
1/2
2/4
4/4
0.5/1
1/1
1/2
16/16
8/8
16/16
4/4
16/16
16/16
16/16
62.5/62.5
62.5/62.5
>250/>250
8/32
2/8
62.5/62.5
32/32
32/32
>250/>250
16/16
16/16
16/16
>250/>250
Xg
Xh
Xi
INHe
8/16
8/16
16/16
>250/>250
Vh
Vi
INHe
a) Values given for 14 days/21 days of incubation; b) nd – not
determined due to low solubility of the compound; c) nt – not
tested; d) the MIC values taken from Waisser et al. (2000); e)
isoniazid (isonicotinhydrazide) used as a reference drug.
a) Values given for 14 days/21 days of incubation; b) nd – not
determined due to low solubility of the compound, c) nt – not
tested; d) the MIC values taken from Waisser et al. (2000); e)
isoniazid (isonicotinhydrazide) used as a reference drug.
with the activity of INH is shown in Tables 4 and 5.
The antimycobacterial activities of the starting 3-
benzyl-2H-1,3-benzoxazine-2,4(3H)-diones were in the
range of 4–16 µmol L−1. The replacement of one oxo
group with a thioxo group significantly increased the
antimycobacterial activity. The further replacement
had a small effect on the activity. Also the intro-
duction of the third sulphur atom by substituting
the phenyl ring with ethoxythiocarbonyl strongly de-
creased the antimycobacterial activity (Waisser et al.,
2009). The MIC values of the sulphur compounds were
generally in the range of 0.5–32 µmol L−1 but the ma-

364
E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Table 6. Activity contribution of the Free–Wilson analyses of of 3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H )-ones and statistical
significance of correlation
∆log(MIC/(µmol L−1))
Parameter
M. tuberculosis
M. avium
M. kansasii
M. kansasii
My 331/88
My 330/88
My 235/80
6509/96
bR1: Br
CH3
F
Cl
OCH3
R2: 4-CH3
4-Cl
–1.333 ( 0.109)
–1.200 ( 0.109)
–1.067 ( 0.109)
–1.267 ( 0.109)
0
–0.180 ( 0.146)
–0.240 ( 0.146)
–0.120 ( 0.146)
–0.180 ( 0.146)
–0.060 ( 0.146)
0.120 ( 0.146)
0.420 ( 0.146)
0.420 ( 0.149)
0
–0.379 ( 0.125)
–0.103 ( 0.121)
–0.767 ( 0.121)
–0.467 ( 0.121)
0
0.00 ( 0.162)
–0.060 ( 0.162)
0.060 ( 0.162)
0.180 ( 0.162)
0.060 ( 0.162)
0.218 ( 0.176)
0.300 ( 0.162).
0.600 ( 0.162)
0
–0.733 ( 0.102)
–0.733 ( 0.102)
–0.933 ( 0.102)
–0.700 ( 0.102)
0
0.120 ( 0.137)
0.180 ( 0.137)
0.060 ( 0.137)
0.360 ( 0.137)
0.180 ( 0.137)
0.660 ( 0.137)
0.840 ( 0.137)
0.780 ( 0.170)
0
–0.700 ( 0.118)
–0.633 ( 0.118)
–1.000 ( 0.118)
–0.967 ( 0.118)
0
0.120 ( 0.159)
0.240 ( 0.159)
0.120 ( 0.159)
0.540 ( 0.159)
0.360 ( 0.159)
0.840 ( 0.159)
0.960 ( 0.159)
0.960 ( 0.159)
0
3-Cl
3,4-Cl2
4-Br
4-F
3-F
3-CF3
4-H
µo
1.213 ( 0.124)
1.549 ( 0.138)
0.98 ( 0.116)
0.803 ( 0.134)
R
s
F
n
0.943
0.230
21.22
45
0.883
0.256
9.12
44
0.926
0.216
15.81
45
0.923
0.251
15.30
45
jority of MICs was in the range of 0.5–4 µmol L−1
.
The following equation illustrates the structure–
activity relationship of 3-phenyl-2H-1,3-benzoxazine-
2,4(3H )-dithiones (VI–X ):
The activities against M. tuberculosis of most of the
compounds tested were comparable with the activity
of INH but the activities against M. kansasii 6509/96
are higher.
The Hansch approach was carried out to study the
relationship between the structure of the compounds
and their activity against M. tuberculosis. The cor-
relation of the logarithm of the minimum inhibitory
concentration for 14 d and 21 d is statistically signifi-
cant so the QSAR study included only the results ob-
tained after 14 d of incubation. The reliability of the
calculation is confirmed by the leave-one-out (LOO)
method.
log(MICM.tub.14d) = – (1.028 0.166)π1 +
+ (1.206 0.302)σ2 – (0.604 0.170)π2 +
+ (0.830 0.127)
(2)
(r = 0.762, Q2 = 0.485, s = 0.366, n = 44, F = 18.52,
Fα=0.05 = 2.84)
Eq. (3) illustrates the structure–activity relation-
ship of 3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H )-
ones (I–V) and 3-phenyl-2H-1,3-benzoxazine-2,4(3H )-
dithiones (VI–X) combined:
Equation 1 illustrates the structure–activity re-
lationship of 3-phenyl-4-thioxo-2H-1,3-benzoxazine-
2(3H )-ones (I–V ):
log(MICM.tub.14d) = – (1.116 0.089)π1 +
+ (1.046 0.219)σ2 – (0.564 0.123)π2 +
+ (0.866 0.089)
(3)
log(MICM.tub.14d) = – (1.202 0.176)π1 +
+ (0.886 0.327)σ2 – (0.522 0.182)π2 +
(r = 0.757, Q2 = 0.529, s = 0.375, n = 89, F = 37.99,
Fα=0.05 = 2.71)
+ (0.898 0.130)
(1)
(r = 0.760, Q2 = 0.486, s = 0.396, n = 45, F = 18.69,
Fα=0.05 = 2.83)
Index 1 represents the parameters of R1 sub-
stituents (in position 6 of the benzoxazine ring) and in-
dex 2 the parameters of R2 substituents on the phenyl
ring. Both the equations have a significance level lower
than 0.05. Both regression equations satisfy the statis-
tical criteria for rejecting the null hypothesis at a sig-
nificance level lower than 0.05. Cross-validation (Q2)
proved the acceptable stability of these QSAR mod-
els. The equations show that activity increases with
the lipophilicity of the substituents. The Free–Wilson
where r is the correlation coefficient, Q2 is the cross-
validation value, s is the standard deviation, n is the
number of compounds used for calculation, F is the
Fischer–Snedecor distribution, F-test, symbol π1 rep-
resents Hansch constants of R1 substituents, π2 Han-
sch constants of R2 substituents, and σ2 Hammett
constants of R2 substituents on the phenyl ring.

E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
365
Table 7. Activity contribution of the Free–Wilson analyses of 3-phenyl-2H-1,3-benzoxazine-2,4(3H )-dithiones and statistical sig-
nificance of correlation
∆log(MIC/(µmol L−1))
Parameter
M. tuberculosis
M. avium
M. kansasii
M. kansasii
My 331/88
My 330/88
My 235/80
6509/96
R1: Br
CH3
–1.037 ( 0.106)
–1.203 ( 0.106)
–0.870 ( 0.106)
–1.203 ( 0.106)
0
–0.366 ( 0.147)
–0.246 ( 0.144)
–0.187 ( 0.147)
–0.126 ( 0.147)
–0.306 ( 0.147)
0.066 ( 0.147)
0.474 ( 0.147)
0.294 ( 0.147)
0
–0.126 ( 0.148)
–0.762 ( 0.148)
–0.291 ( 0.148)
–0.292 ( 0.148)
0
0.120 ( 0.186)
0.360 ( 0.186)
–0.225 ( 0.194)
0.360 ( 0.186)
0.360 ( 0.186)
0.240 ( 0.186)
0.450 ( 0.194).
0.660 ( 0.186)
0
–0.688 ( 0.119)
–0.721 ( 0.119)
–0.621 ( 0.119)
–0.755 ( 0.119)
0
0.145 ( 0.157)
0.206 ( 0.157)
0.206 ( 0.157)
0.266 ( 0.157)
0.206 ( 0.157)
0.387 ( 0.157)
0.750 ( 0.164)
0.686 ( 0.157)
0
–0.587 ( 0.126)
–0.620 ( 0.126)
–0.787 ( 0.126)
–0.920 ( 0.126)
0
0.004 ( 0.167)
0.304 ( 0.167)
0.124 ( 0.167)
0.244 ( 0.167)
0.304 ( 0.167)
0.604 ( 0.167)
0.825 ( 0.175)
0.964 ( 0.167)
0
F
Cl
OCH3
R2: 4-CH3
4-Cl
3-Cl
3.4-Cl2
4-Br
4-F
3-F
3-CF3
4-H
µo
1.228 ( 0.124)
1.200 ( 0.186)
1.071 ( 0.152)
0.879 ( 0.162)
R
s
F
n
0.940
0.218
19.90
44
0.852
0.275
6.35
42
0.869
0.232
7.73
43
0.904
0.247
11.17
43
method modified by Fujita–Ban was used to investi-
gate the activity contribution (Tables 6 and 7). The
alkyl derivatives are proposed for future screening be-
cause of the decrease in the activity with an increasing
value of σ2 constants and the Free–Wilson analysis re-
sults.
Vincent, V., Marchal, G., Dupont, B., Bouchaud, O., Valeyre,
D., & Lortholary, O. (2004). Tuberculosis in HIV-infected pa-
tients: a comprehensive review. Clinical Microbiology and In-
fection, 10, 388–398. DOI: 10.1111/j.1469-0691.2004.00758.x.
Dye, C. (2009). Doomsday postponed? Preventing and revers-
ing epidemics of drug-resistant tuberculosis. Nature Reviews
Microbiology, 7, 81–87. DOI: 10.1038/nrmicro2048.
Free, S. M., & Wilson, J. W. (1964). A mathematical contri-
bution to structure-activity studies. Journal of Medicinal
Chemistry, 7, 395–399. DOI: 10.1021/jm00334a001.
Conclusions
Fujita, T.,
& Ban, T. (1971). Structure-activity relation.
The derivatives of 3-phenyl-4-thioxo-2H-1,3-benz-
oxazine-2(3H )-one (I–V ) and 3-phenyl-2H-1,3-benz-
oxazine-2,4(3H )-dithione (VI–X ) possess a biological
activity similar to INH, the standard used for compar-
ison. The replacement of one oxo group with a thioxo
group strongly increased the antimycobacterial activ-
ity. The replacement of a further oxo group had a small
effect on the activity. 3-(4-Chlorophenyl)-6-methyl-
4-thioxo-2H-1,3-benzoxazine-2(3H )-one (IIc) is the
most promising compound, as it shows the best activ-
ity against all the mycobacterial strains under exam-
ination. This compound has been selected for future
investigation.
3. Structure-activity study of phenethylamines as sub-
strates of biosynthetic enzymes of sympathetic transmit-
ters. Journal of Medicinal Chemistry, 14, 148–152. DOI:
10.1021/jm00284a016.
Golbraikh, A., & Tropsha, A. (2002). Beware of q2! Journal
of Molecular Graphics and Modelling, 20, 269–276. DOI:
10.1016/S1093-3263(01)00123-1.
Gupta, R. A., Gupta, A. K., Soni, L. K., & Kaskhedikar, S. G.
(2009). Study of physicochemical properties–antitubercular
activity relationship of naphtalene-1,4-dione analogs:
QSAR approach. Chemical Papers, 63, 723–730. DOI:
10.2478/s11696-009-0080-0.
Hansch, C., & Leo, A. J. (1979). Substituent constants for cor-
relation analysis in chemistry and biology. New York, NY,
USA: Wiley.
Hlasta, D. J., Demers, J. P, Foleno, B. D, Frago-Spano, S. A.,
Guan, J., Hilliar, J. J., Macielag, M. J., Ohemeng, K. A.,
Sheppard, C. M., Sui, Z., Webb, G. C., Weidner-Wells, M.
A., Werblood, H., & Barrett, J. F. (1998). Novel inhibitors
of bacterial two-component systems with gram positive an-
tibacterial activity: Pharmacofore identification based on the
screening hit closantel. Bioorganic & Medicinal Chemistry
Letters, 8, 1923–1928. DOI: 10.1016/S0960-894X(98)00326-
6.
A
Acknowledgements. This work was financially supported by
project No. MSM 0021620822 of the Ministry of Education
of the Czech Republic, Grant Agency of Charles University
(GAUK 11809/2009/B-CH/FAF) and by grant No. SVV-2010-
261-001.
References
Aaron, L., Saadoun, D., Calatroni, I., Launay, O., Mémain, N.,

366
E. Petrlíková et al./Chemical Papers 65 (3) 352–366 (2011)
Li, X., Liu, N., Zhang, H., Knudson, S. E., Slayden, R.
A., Tonge, P. J. (2010). Synthesis and SAR stud-
ies of 1,4-benzoxazine MenB inhibitors: Novel antibac-
terial agents against Mycobacterium tuberculosis. Bioor-
ganic & Medicinal Chemistry Letters, 20, 6306–6309. DOI:
10.1016/j.bmcl.2010.08.076.
Macielag, M. J., Demers, J. P., Fraga-Spano, S. A., Hlasta, D.
J., Johnson, S. G., Kanojia, R. M., Russell, R. K., Sui, Z.,
Weidner-Wells, M. A., Werblood, H., Foleno, B. D., Gold-
schmidt, R. M., Loeloff, M. J., Webb, G. C., & Barrett,
J. F. (1998). Substituted salicylanilides as inhibitors of two-
component regulatory systems in bacteria. Journal of Medic-
inal Chemistry, 41, 2939–2943. DOI: 10.1021/jm9803572.
Matyk, J., Waisser, K., Dražková, K., Kuneš, J., Klimešová, V.,
Palát, K., Jr., Kaustová, J. (2005). Heterocyclic isosters of
antibacterial salicylanilides. II Farmaco, 60, 399–408. DOI:
10.1016/j.farmac.2005.02.002.
Tortoli, E. (2009). Clinical manifestations of nontuberculous
mycobacteria infections. Clinical Microbiology and Infection,
15, 906–910. DOI: 10.1111/j.1469-0691.2009.03014.x.
van den Boogaard, J., Kibiki, G. S., Kisanga, E. R., Boeree, M.
J., & Aarnoutse, R. E. (2009). New drugs against tubercu-
losis: Problems, progress, and evaluation of agents in clinical
development. Antimicrobial Agents and Chemotherapy, 53,
849–862. DOI: 10.1128/AAC.00749-08.
Wagner, G., Singer, D., & Weuffen, W. (1966). Studies on
2-hydroxythiobenzamide and 2-hydroxythiobenzanilide. 1.
Synthesis of the compounds. Pharmazie, 21, 161–166.
Waisser, K., Čižmárik, J., Holý, P., Petrlíková, E., Kuneš, J.,
& Kaustová, J. (2009). Antimycobacterial 3-(4-ethoxythio-
carbonylphenyl)-4-thioxo-2H-1,3-benzoxazine-2(3H )-ones
and 3-(4-ethoxythiocarbonylphenyl)-2H-1,3-benzoxazine-
2,4(3H )-dithiones. Acta Facultatis Pharmaceuticae Univer-
sitatis Comenianae, 56, 171–179.
&
Naidoo, K., Naidoo, K., Padayatchi, N., & Karim, Q. A.
(2011). HIV-associated tuberculosis. Clinical and Develop-
mental Immunology, 2011, Article ID 585919, 8 pages. DOI:
10.1155/2011/585919.
Nemeček, P., Ďurčeková, T., Mocák, J., & Waisser, K. (2009).
Chemometrical analysis of computed QSAR parameters and
their use in biological activity prediction. Chemical Papers,
63, 84–91. DOI: 10.2478/s11696-008-0089-9.
Waisser, K., Gregor, J., Kubicová, L., Klimešová, V., Kuneš,
J., Macháček, M.,
& Kaustová, J. (2000). New groups
of antimycobacterial agents: 6-chloro-3-phenyl-4-thioxo-2H-
1,3-benzoxazine-2(3H )-ones and 6-chloro-3-phenyl-2H-1,3-
benzoxazine-2,4(3H )-dithiones. European Journal of Medici-
nal Chemistry, 35, 733–741. DOI: 10.1016/S0223-5234(00)00
174-4.
Waisser, K., Hladůvková, J., Holý, P., Macháček, M., Karajan-
nis, P., Kubicová, L., Klimešová, V., Kuneš, J., & Kaustová,
J. (2001a). 2H-1,3-benzoxazine-2,4(3H )-diones substituted in
position 6 as antimycobacterial agents. Chemical Papers, 55,
323–334.
Waisser, K, Hladůvková, J, Kuneš, J, Kubicová, L, Klimešová,
V, Karajannis, P, & Kaustová, J. (2001b). Synthesis and an-
timycobacterial activity of salicylanilides substituted in po-
sition 5. Chemical Papers, 55, 121–129.
Waisser, K., Matyk, J., Divišová, H., Husáková, P., Kuneš, J.,
Klimešová, V., Kaustová, J., M¨ollmann, U., Dahse, H.-M., &
Miko, M. (2006). The oriented development of antitubercu-
lotics: Salicylanilides. Archiv der Pharmazie, 339, 616–620.
DOI: 10.1002/ardp.200600093.
O’Brien, R. J., & Nunn, P. P. (2001). The need for new drugs
against tuberculosis. American Journal of Respiratory and
Critical Care Medicine, 163, 1055–1058.
Petrlíková, E., Waisser, K., Jílek, P., & Dufková, I. (2010). An-
tibacterial activity of N-nenzylsalicylthioamides, Folia Mi-
crobiologica, 55, 418–421. DOI: 10.1007/s12223-010-0070-1.
Petrlíková E., Waisser K., Palát, K., Kuneš, J., & Kaustová
J. (2011). A new group of potential antituberculotics: N-(2-
pyridylmethyl)salicylamides and N-(3-pyridylmethyl)salicyl-
amides, Chemical Papers, 65, 52–59. DOI: 10.2478/s11696-
010-0084-9.
Saeed, A., & Ashraf, Z. (2008). Synthesis of some 3-aryl-1H-
isochromene-1-thiones. Journal of Heterocyclic Chemistry,
45, 679–682. DOI: 10.1002/jhet.5570450307.
Schroeder, E. K., de Souza, O. N., Santos, D. S., Blan-
chard, J. S., & Basso, L. A. (2002). Drugs that inhibit
mycolic acid biosynthesis in mycobacterium tuberculosis.
Current Pharmaceutical Biotechnology, 3, 197–225. DOI:
10.2174/1389201023378328.