Relationship between the structure and antimycobacterial activity of substituted salicylanilides

Article abstract of DOI:10.1002/ardp.200390004

A series of 143 salicylanilides substituted in positions 4 and 5 and in positions 3′ and 4′ was synthesized. The compounds were evaluated for in vitro antimycobacterial activity against Mycobacterium tuberculosis, Mycobacterium kansasii, and Mycobacterium

Full text of DOI:10.1002/ardp.200390004

Arch. Pharm. Pharm. Med. Chem. 2003, 1, 53–71
Antimycobacterial Activity of Salicylanilides 53
Karel Waissera,
Relationship between the Structure and
Antimycobacterial Activity of Substituted
Salicylanilides
Otakar Bureša,
Pavel Holy´ a,
Jišrí Kuneša,
Radek Oswalda,
Lucie Jiráskováa,
Milan Poura,
A series of 143 salicylanilides substituted in positions 4 and 5 and in positions 3Ј and
4Ј was synthesized. The compounds were evaluated for in vitro antimycobacterial
activity against Mycobacterium tuberculosis, Mycobacterium kansasii, and Myco-
bacterium avium.To describe the structure-antimycobacterial activity relationships
(QSARs), an approach based on the combination of the Free-Wilson and Hansch
methods was employed (the substituent constants were used in the case of the sub-
stituents on the phenyl ring; indicator parameters were used for the substituents on
the acyl moiety).The relationships between the antimycobacterial activity and physi-
co-chemical parameters of all substituents were also explored.The quadratic repre-
sentation of lipophilicity parameters did not lead to significant correlations.
Všera Klimešováa,
Lenka Kubicováa,
Jarmila Kaustováb
a
Department of Inorganic
and Organic Chemistry,
Faculty of Pharmacy,
Heyrovského 1203,
CZ 500 05 Hradec Králové,
Czech Republic
National Reference
b
Key Words: Salicylanilides; Antimycobacterial activity; Structure-activity relation-
ships; Tuberculostatics; Atypical mycobacterial strains
Laboratory for
Mycobacterium kansasii,
Institute of Hygiene,
Partyzánské nám. 7,
CZ 728 892 Ostrava,
Czech Republic
Received: April 25, 2002 [FP692]
electron-accepting properties of the substituents [5, 6].
When salicylanilides were substituted in position 5, the
Introduction
The search for new antimycobacterial compounds is one
of the most challenging tasks of current medicinal chem-
istry. In particular, a number of new broad-spectrum an-
timycobacterial substances from the class of organic
sulphur compounds were synthesized by Mayer and co-
workers and are reviewed in our previous paper [1]. Our
own research on antimycobacterial salicylanilides was
preceded by a review on the biological activity of these
compounds [2].The study of antimycobacterial activity of
salicylanilides is currently of great importance, because
salicylanilides are inhibitors of the bacterial two-compo-
nent system [3] which is also found in mycobacteria [4].
These substances are convenient starting materials in
the synthesis of antimycobacterial 3-aryl-2H-1,3-ben-
zoxazin-2,4(3H)-diones, and also possess antimyco-
bacterial activity. However, salicylanilides are unlikely to
be effective chemotherapeutic agents themselves due to
their effect on mitochondrial respiration [3].We have pre-
viously found relatively simple SARs in the group of sali-
cylanilides with no substitution in the acyl moiety, where
the activity rises with increasing hydrophobicity and
influence of the substituents on the phenyl ring in the ani-
lide part of the molecule was similar. However, the influ-
ence of the substituents in the acyl moiety was quite
complex [7]. For this reason, we set out to synthesize a
large series of compounds.The goal of this work was to
assess the relationships between the antimycobacterial
activity against Mycobacterium tuberculosis, M.kansasii
and M.avium, and the structure of salicylanilides bearing
substituents in positions 4 and 5, and on the phenyl ring
in the anilide part of their molecules. The influence of
substituents in position 4 has not yet been studied.
Chemistry
The starting salicylanilides (1–10) were prepared by the
treatment of substituted salicylic acids with the appropri-
ate anilines in chlorobenzene.
Microbiology
Compounds 1–10 were tested for their in vitro antimyco-
bacterial activity against Mycobacterium tuberculosis
CNCTC My 331/88, Mycobacterium kansasii CNCTC
My 235/80, and Mycobacterium avium CNCTC My
330/88 obtained from the Czech National Collection of
Type Cultures (CNCTC).
Correspondence: Karel Waisser, Department of Inorganic
and Organic Chemistry, Faculty of Pharmacy, Heyrovského
1203, CZ 500 05 Hradec Králové, Czech Republic.
Phone: +420 49 5067111, fax: +420 49 5210002, e-mail:
waisser@faf.cuni.cz.
© 2003 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
0365-6233/01/0053 $ 17.50+.50/0

54 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Standard strains of mycobacteria from the Czech
National Collection of Type Cultures (CNCTC) were em-
ployed.In several cases, the minimum inhibitory concen-
tration could not be determined due to the limited solubil-
ity of the compounds.In general, the results suggest that
the compounds under study can be considered as pro-
spective broad-spectrum antimycobacterial sub-
stances.
Calculation
To set up the regression equations, we employed the
“Multireg” programme developed by Klemera for Micro-
soft Excel. The values of the substituent constants and
molecular refraction were taken from the literature [8, 9],
with the exception of the Hammett constant for the thio-
amide group, which was taken from the original paper
[10]. There are no significant intercorrelations between
different sets of parameters.
In studying the relationship between the structure and
antimycobacterial activity, we followed up on the conclu-
sions drawn in our previous work [7].The Hammett con-
stants σ of the substituents on the phenyl ring served as
one parameter, and their hydrophobic substituent con-
stants π were used as the second parameter. However,
the influence of the substituents from the acyl part of the
molecule appeared to be more complex, and hence we
expressed them with indicator parameters (In). The
QSAR study of the antimycobacterial activity of salicyl-
Results and discussion
The chemical and physical data of the new salicylani-
lides are given in Table 1. In the spectra of the salicyl-
anilides (1–10), the amide ν(C=O) vibrations are clearly
observed. Compounds described previously [5–7] are
not included in the Table. NMR spectra are given in
Table 2.
Scheme 1. Preparation of the salicylanilides
Scheme 2. Overview of the structures of the salicylanilides.

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 55
Table 1. Characteristics of the compounds.
Compound R1
R2
Formula
ν C=O
Mp [°C]
Mr
[cm–1]
Found
Ref.
2 a
2 b
2 c
2 d
2 e
2 f
4-Cl
H
C13H10ClNO2
247.7
C14H12ClNO2
261.7
C13H9Cl2NO2
282.12
C13H9Cl2NO2
282.12
C13H8Cl3NO2
316.57
C13H9BrClNO2
326.6
C13H9FClNO2
265.7
C13H9FClNO2
265.7
C14H9ClF3NO2
315.67
C13H9ClN2O4
292.7
C15H15ClN2O2
290.8
C14H12ClNO3
277.7
C16H14ClNO4
319.74
C14H9ClN2O2
272.69
C14H13NO2
227.26
C15H15NO2
241.28
C14H12ClNO2
261.7
C14H12ClNO2
261.7
C14H11Cl2NO2
296.15
C14H12BrNO2
306.15
C14H12FNO2
245.25
C14H12FNO2
245.25
C15H12F3NO2
295.26
C14H12N2O4
272.26
1634
218–220
220–222
222–223
216–218
221–223
220–222
205–207
226–227
213–216
225–226
224–234
205–207
216–218
238–240
186–188
202–204
217–219
199–201
208–210
225–227
176–179
137–139
229–231
216–218
201–203
217–220 [12]
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-CH3
4-CH3
4-Cl
1628
–
1611
–
3-Cl
1619
–
3,4-Cl2
4-Br
1617
221–222 [13]
1606
–
2 g
2 h
2 i
4-F
1620
–
3-F
1615
–
4-CF3
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
H
1607
–
2 k
2 l
1641
–
1616
–
2 m
2 n
2 o
3 a
3 b
3 c
3 d
3 e
3 f
1617
–
1648;1685
–
1648
(CN) 2227
1622
–
190–191 [14]
4-CH3 4-CH3
4-CH3 4-Cl
1603
1623
1624
1626
1622
1622
1617
1618
1631
1641
166–167 [14]
–
4-CH3 3-Cl
–
4-CH3 3,4-Cl2
4-CH3 4-Br
4-CH3 4-F
–
–
3 g
3 h
3 i
–
4-CH3 3-F
–
4-CH3 4-CF3
4-CH3 3-NO2
4-CH3 4-N(CH3)2
–
3 k
3 l
120–121 [14]
–
C16H18N2O2
270.33

56 Waisser et al.
Table 1. continued.
Compound R1
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
R2
Formula
ν C=O
Mp [°C]
Mr
[cm–1]
Found
Ref.
3 m
3 n
3 o
4 a
4 b
4 c
4 d
4 e
4 f
4-CH3
4-CH3
4-CH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
5-Br
4-OCH3
4-COOEt
4-CN
C15H15NO3
257.28
C17H17NO4
299.32
C15H12N2O2
252.27
C14H13NO3
243.26
C15H15NO3
257.28
C14H12ClNO3
277.7
C14H12ClNO3
277.7
C14H11Cl2NO3
312.15
C14H12BrNO3
322.15
C14H12FNO3
261.25
C14H12FNO3
261.25
C15H12F3NO3
311.26
C14H12N2O5
288.26
C14H12N2O5
288.26
C15H15NO4
273.29
C17H17NO5
315.32
C15H12N2O3
268.27
C14H9BrF3NO2
360.13
C16H14BrNO4
364.19
C14H9BrN2O2
317.14
C14H9ClF3NO2
315.67
C16H14ClNO4
319.74
C14H9ClN2O2
272.69
C14H9F4NO2
299.22
1628
183–185
194–196
212–213
150–152
146–147
184–186
165–167
203–205
185–187
174–176
182–185
145–146 [14]
1617;1713
–
1641
(CN) 2227
1621
–
H
152–154 [15]
4-CH3
4-Cl
1614
1617
151–152 [16]
–
3-Cl
1618
–
3,4-Cl2
4-Br
1622
–
1617
–
4 g
4 h
4 i
4-F
1618
–
3-F
1622
–
4-CF3
4-NO2
3-NO2
4-OCH3
4-COOEt
4-CN
1621
–
4 j
1626
222–224
189–191
163–165
193–194
218–220
202–203
235–236.5
256–257
217–218
212–214
242–243.5
222–223
193–195
223–224 [16]
4 k
4 m
4 n
4 o
5 i
1643
194–195 [16]
1616
164–165 [16]
1611; 1711
–
–
–
–
–
–
–
–
–
–
1647
(CN) 2224
1635
4-CF3
4-COOEt
4-CN
5 n
5 o
6 i
5-Br
1648; 1686
5-Br
1637
(CN) 2229
1629
5-Cl
4-CF3
4-COOEt
4-CN
6 n
6 o
7 i
5-Cl
1632; 1717
5-Cl
1636
(CN) 2231
1630
5-F
4-CF3
4-COOEt
7 n
5-F
C16H14FNO4
303.28
1640; 1690

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 57
Table 1. continued.
Compound R1
R2
Formula
ν C=O
Mp [°C]
Mr
[cm–1]
Found
Ref.
7 o
8 a
8 b
8 c
8 d
8 e
8 f
5-F
4-CN
H
C14H9FN2O2
256.23
C14H13NO2
227.26
C15H15NO2
241.28
C14H12ClNO2
261.7
C14H12ClNO2
261.7
C14H11Cl2NO2
296.15
C14H12BrNO2
306.15
C14H12FNO2
245.25
C14H12FNO2
245.25
C15H12F3NO2
295.26
C14H12N2O4
272.26
C14H12N2O4
272.26
C16H18N2O2
270.33
C15H15NO3
257.28
C17H17NO4
299.32
C15H12N2O2
252.27
C14H13NO3
243.26
C15H15NO3
257.28
C14H12ClNO3
277.7
C14H12ClNO3
277.7
C14H11Cl2NO3
312.15
C14H12BrNO3
322.15
C14H12FNO3
261.25
C14H12FNO3
261.25
1643
(CN) 2229
1629
218–219
159–161
189–191
197–198
170–172
210–212
204–206
166–168
183–184
207–208
234–236
197–199
197–199
182–184
300–302
140–142
167–169
188–190
209–210
162–164
204–206
223–225
181–183
178–180
207
–
5-CH3
160–161 [17]
5-CH3
4-CH3
4-Cl
1631
1633
167–168 [14]
5-CH3
164–165 [17]
5-CH3
3-Cl
1646
–
5-CH3
3,4-Cl2
4-Br
1644
212–213 [18]
5-CH3
1632
210–211 [19]
8 g
8 h
8 i
5-CH3
4-F
1634
–
5-CH3
3-F
1634
–
5-CH3
4-CF3
4-NO2
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
H
1643
–
8 j
5-CH3
1636
236–237 [14]
8 k
8 l
5-CH3
1652
120–121 [14]
5-CH3
1616
–
8 m
8 n
8 o
9 a
9 b
9 c
9 d
9 e
9 f
5-CH3
1629
185–186 [19]
5-CH3
1644; 1698
–
5-CH3
1662
(CN) 2227
1636
–
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
–
4-CH3
4-Cl
1634
1637
1642
1641
1637
1641
1642
1636
–
–
3-Cl
–
3,4-Cl2
4-Br
–
214–215 [19]
9 g
9 h
9 i
4-F
–
–
–
3-F
4-CF3
C15H12F3NO3
311.26

58 Waisser et al.
Table 1. continued.
Compound R1
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
R2
Formula
ν C=O
Mp [°C]
Mr
[cm–1]
Found
Ref.
–
9 j
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
4-NO2
C14H12N2O5
288.26
C14H12N2O5
288.26
C15H15NO4
273.29
C17H17NO5
315.32
1656
1642
243–244
219–221
188–190
196–197
232–233
9 k
9 m
9 n
9 o
3-NO2
–
4-OCH3
4-COOEt
4-CN
1634
–
1637; 1685
–
C15H12N2O3
268.27
1644
–
(CN) 2224
Compounds described previously [5–7] were not included. For other compounds, literature references are given.
Table 2. 1H and 13C NMR spectra of the new salicylanilides.
Compound
NMR, δ
2 a
1H NMR (300 MHz, DMSO) δ 12.11 (bs, 1 H, OH), 10.37 (s, 1 H, NH), 7.97–7.91 (m, 1 H, H6), 7.72–7.65
(m, 2 H, H2Ј, H6Ј), 7.41–7.29 (m, 2 H, H3Ј, H5Ј), 7.17–7.07 (m, 1 H, H4Ј), 7.06–6.98 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ 165.7, 159.2, 138.3, 137.6, 131.1, 129.0, 124.5, 121.1, 119.4, 117.5,
117.0
2 b
1H NMR (300 MHz, DMSO) δ 12.21 (bs, 1 H, OH), 10.31 (s, 1 H, NH), 7.98–7.93 (m, 1 H, H6), 7.61–7.53
(m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.20–7.12 (m, AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.07–6.98 (m, 2 H, H3, H5), 2.27 (s,
3 H, CH3)
13C NMR (75 MHz, DMSO) δ 165.7, 159.4, 137.6, 135.7, 133.6, 130.9, 129.4, 121.2, 119.4, 117.1,
117.0, 20.7
2 c
2 d
1H NMR (300 MHz, DMSO) δ 11.94 (bs, 1 H, OH), 10.43 (s, 1 H, NH), 7.89 (d, 1 H, J = 8.24 Hz, H6),
7.78–7.67 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.45–7.35 (m, AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.08–6.98 (m, 2 H, H3,
H5)
13C NMR (75 MHz, DMSO) δ 165.6, 158.9, 137.6, 137.4, 131.1, 128.9, 128.0, 122.5, 119.5, 117.7, 117.0
1H NMR (300 MHz, DMSO) δ 11.86 (bs, 1 H, OH), 10.45 (s, 1 H, NH), 7.91 (t, 1 H, J = 1.92 Hz, H2Ј),
7.90–7.85 (m, 1 H, H6), 7.59 (ddd, 1 H, J = 8.24 Hz, J = 1.92 Hz, J = 1.10 Hz, H6Ј), 7.38 (t, 1 H, J = 8.24
Hz, H5Ј), 7.18 (ddd, 1 H, J = 8.24 Hz, J = 1.92 Hz, 1.10 Hz, H4Ј), 7.06–7.00 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ 165.6, 158.7, 139.9, 137.6, 133.3, 131.2, 130.6, 124.0, 120.3, 119.5,
119.2, 117.9, 116.9
2 e
1H NMR (300 MHz, DMSO) δ 11.78 (bs, 1 H, OH), 10.50 (s, 1 H, NH), 8.09 (d, 1 H, J = 2.19 Hz, H2Ј), 7.84
(d, 1 H, J = 8.52 Hz, H6), 7.65 (dd, 1 H, J = 8.79 Hz, J = 2.19 Hz, H5Ј), 7.59 (d, 1 H, J = 8.79 Hz, H6Ј),
7.05–6.99 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ 165.6, 158.5, 138.6, 137.6, 131.3, 131.2, 130.8, 125.8, 121.9, 120.8,
119.5, 118.0, 116.9
2 f
1H NMR (300 MHz, DMSO) δ 11.94 (bs, 1 H, OH), 10.43 (s, 1 H, NH), 7.92–7.86 (m, 1 H, H6), 7.72–7.63
(m, AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.59–7.50 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.07–6.98 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ 165.3, 158.6, 137.5, 137.3, 131.5, 130.9, 122.6, 119.2, 117.5, 116.7, 115.9
1H NMR (300 MHz, DMSO) δ 12.02 (bs, 1 H, OH), 10.40 (s, 1 H, NH), 7.92 (d, 1 H, J = 8.52 Hz, H6),
7.76–7.66 (m, 2 H, H2Ј, H6Ј), 7.23–7.16 (m, 2 H, H3Ј, H5Ј), 7.05–7.00 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ 165.7, 160.5 and 157.3 (J = 241.1 Hz), 159.3, 137.6, 134.6 and 134.6 (J =
2.9 Hz), 131.0, 123.1 and 123.0 (J = 8.0 Hz), 119.4, 117.3, 117.0, 115.7 and 115.4 (J = 23.4 Hz)
2 g

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 59
Table 2. continued.
Compound
NMR, δ
2 h
1H NMR (300 MHz, DMSO) δ 11.88 (bs, 1 H, OH), 10.48 (s, 1 H, NH), 7.90–7.85 (m, 1 H, H6), 7.70 (dt,
1 H, J = 11.54 Hz, J = 2.20 Hz, H6Ј), 7.49–7.33 (m, 2 H, H2Ј, H5Ј), 7.06–6.91 (m, 3H, H3, H5, H4Ј)
13C NMR (75 MHz, DMSO) δ 165.6, 163.9 and 160.7 (J = 241.3 Hz), 158.6, 140.2 and 140.1 (J = 10.9
Hz), 137.5, 131.3, 130.7 and 130.5 (J = 9.4 Hz), 119.5, 118.0, 116.9, 116.6 and 116.5 (J = 2.6), 110.9
and 110.7 (J = 20.9 Hz), 107.7 and 107.4 (J = 26.0 Hz)
2 i
1H NMR (300 MHz, DMSO) δ 11.83 (bs, 1 H, OH), 10.61 (s, 1 H, NH), 7.96–7.85 (m AAЈ, BBЈ overlapped,
3H, H6, H3Ј, H5Ј), 7.75–7.68 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.07–7.00 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ 165.7, 158.5, 142.2, 137.6, 126.3 (q, J = 4.0 Hz), 124.5 (q, J = 271.4 Hz),
122.7, 124.2 (q, J = 32.1 Hz), 120.6, 119.5, 118.2, 116.9
2 k
1H NMR (300 MHz, DMSO) δ 11.76 (bs, 1 H, OH), 10.70 (s, 1 H, NH), 8.75 (t, 1 H, J = 2.19 Hz, H2Ј),
8.07–8.01 (m, 1 H, H6Ј), 8.00–7.94 (m, 1 H, H4Ј), 7.90–7.84 (m, 1 H, H6), 7.64 (t, 1 H, J = 8.24 Hz, H5Ј),
7.08–7.00 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ 165.9, 158.6, 148.1, 139.7, 137.7, 131.3, 130.4, 126.7, 119.5, 118.8,
118.0, 116.9, 114.8
2 l
1H NMR (300 MHz, DMSO) δ 12.5 (bs, 1 H, OH), 10.24 (s, 1 H, NH), 8.01–7.96 (m, 1 H, H6), 7.51–7.43
(m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.04–6.97 (m, 2 H, H3, H5), 6.76–6.69 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 2.87 (s,
6 H, CH3)
13C NMR (75 MHz, DMSO) δ 165.7, 160.2, 148.0, 137.6, 130.4, 127.4, 122.8, 119.1, 117.1, 116.3,
112.6, 40.6
2 m
1H NMR (300 MHz, DMSO) δ 12.31 (bs, 1 H, OH), 10.29 (s, 1 H, NH), 7.99–7.94 (m, 1 H, H6), 7.63–7.54
(m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.04–6. 99 (m, 2 H, H3, H5), 6.97–6.90 (m AAЈ, BBЈ, H3Ј, H5Ј), 3.74 (s, 3 H,
OCH3)
13C NMR (75 MHz, DMSO) δ 165.8, 159.8, 156.3, 137.6, 131.1, 130.7, 122.9, 119.3, 117.1, 116.7,
114.1, 55.4
2 n
2 o
3 a
1H NMR (300 MHz, DMSO) δ 11.83 (bs, 1 H, OH), 10.55 (s, 1 H, NH), 7.96–7.78 (m, 5 H, H6, H2Ј, H3Ј,
H5Ј, H6Ј), 7.04–6.96 (m, 2 H, H3, H5), 4.25 (q, 1 H, J = 7.14 Hz, CH2), 1.27 (t, 3 H, J = 7.15 Hz, CH3)
13C NMR (75 MHz, DMSO) δ165.6, 165.5, 158.5, 142.9, 137.6, 131.4, 130.4, 125.2, 120, 119.6, 118.2,
116.9, 60.7, 14.4
1H NMR (300 MHz, DMSO) δ 11.76 (bs, 1 H, OH), 10.62 (s, 1 H, NH), 7.94–7.76 (m, 5 H, H6, H2Ј, H3Ј,
H5Ј, H6Ј), 7.05–6.99 (m, 2 H, H3, H5)
13C NMR (75 MHz, DMSO) δ165.6, 158.3, 142.8, 137.6, 133.4, 131.5, 120.6, 119.6, 119.2, 118.5, 116.9,
105.9
1H NMR (300 MHz, DMSO) δ 11.95 (bs, 1 H, OH), 10.34 (bs, 1 H, NH), 7.93–7.88 (m, 1 H, H6), 7.74–7.65
(m, 2 H, H2Ј, H6Ј), 7.40–7.32 (m, 2 H, H3Ј, H5Ј), 7.18–7.08 (m, 1 H, H4Ј), 6.82–6.74 (m, 2 H, H3, H5),
2.29 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.1, 159.3, 144.6, 138.3, 129.0, 129.0, 124.4, 121.3, 120.3, 117.7,
114.3, 21.3
3 b
3 c
1H NMR (300 MHz, DMSO) δ 12.04 (bs, 1 H, OH), 10.26 (s, 1 H, NH), 7.90 (d, 1 H, J = 8.51 Hz, H6),
7.60–7.52 (m, AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.20–7.12 (m, AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.80–6.73 (m, 2 H, H3,
H5), 2.29 (s, 3 H, CH3), 2.27 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.1, 159.5, 144.6, 135.7, 133.5, 129.3, 128.8, 121.4, 120.2, 117.7,
114.1, 21.3, 20.8
1H NMR (300 MHz, DMSO) δ 11.81 (bs, 1 H, OH), 10.41 (s, 1 H, NH), 7.86 (d, 1 H, J = 8.51 Hz, H6),
7.78–7.69 (m, AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.46–7.36 (m, AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.82–6.74 (m, 2 H, H3,
H5), 2.28 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.1, 159.1, 144.7, 137.4, 129.1, 128.8, 128.0, 122.7, 120.3, 117.7,
114.4, 21.3

60 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Table 2. continued.
Compound
NMR, δ
3 d
3 e
3 f
1H NMR (300 MHz, DMSO) δ 11.72 (bs, 1 H, OH), 10.42 (s, 1 H, NH), 7.91 (t, 1 H, J = 2.0 Hz, H2Ј), 7.85
(d, 1 H, J = 8.52 Hz, H6), 7.63–7.57 (m, 1 H, H6Ј), 7.38 (t, 1 H, J = 8.1 Hz, H5Ј), 7.20–7.15 (m, 1 H, H4Ј),
6.82–6.74 (m, 2 H, H3, H5), 2.29 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.1, 158.9, 144.8, 140.0, 133.2, 130.6, 129.2, 124.0, 120.5, 120.4,
119.4, 117.7, 114.6, 21.3
1H NMR (300 MHz, DMSO) δ 11.63 (bs, 1 H, OH), 10.48 (s, 1 H, NH), 8.10 (d, 1 H, J = 2.34 Hz, H2Ј), 7.82
(d, 1 H, J = 7.69 Hz, H6), 7.66 (dd, 1 H, J = 8.79 Hz, J = 2.34 Hz, H6Ј), 7.58 (d, 1 H, J = 8.79 Hz, H5Ј),
6.82–6.74 (m, 2 H, H3, H5), 2.28 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.0, 158.8, 144.8, 138.7, 131.2, 130.8, 129.2, 125.7, 122.1, 120.9,
120.4, 117.7, 114.6, 21.3
1H NMR (300 MHz, DMSO) δ 11.63 (bs, 1 H, OH), 10.40 (s, 1 H, NH), 7.88–7.83 (m, 1 H, H6), 7.71–7.65
(m, AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.56–7.50 (m, AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.81–6.75 (m, 2 H, H3, H5), 2.28 (s,
3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.0, 159.1, 144.7, 137.8, 131.8, 129.1, 123.1, 120.3, 117.7, 116.1,
114.5, 21.4
3 g
3 h
1H NMR (300 MHz, DMSO) δ 11.94 (bs, 1 H, OH), 10.35 (s, 1 H, NH), 7.88 (d, 1 H, J = 8.52 Hz, H6),
7.76–7.65 (m, 2 H, H2Ј, H6Ј), 7.26–7.14 (m, 2 H, H3Ј, H5Ј), 6.82–6.74 (m, 2 H, H3, H5), 2.29 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.2, 160.5 and 157.3 (J = 241.1 Hz), 159.4, 144.7, 134.7 and 134.6 (J =
2.9 Hz), 128.9, 123.3 and 123.2 (J = 8.0 Hz), 120.2, 117.7, 115.7 and 115.4 (J = 22.0 Hz), 114.1, 21.3
1H NMR (300 MHz, DMSO) δ 11.72 (bs, 1 H, OH), 10.45 (s, 1 H, NH), 7.88–7.83 (m, 1 H, H6), 7.74–7.66
(m, 1 H, H6Ј), 7.50–7.32 (m, 2 H, H2Ј, H5Ј), 7.06–6.90 (m, 1 H, H4Ј), 6.83–6.75 (m, 2 H, H3, H5), 2.29 (s,
3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.0, 163.9 and 160.7 (J = 241.4 Hz), 158.8, 144.7, 140.3 and 140.1 (J =
11.2 Hz), 130.6 and 130.5 (J = 9.4 Hz), 129.2, 120.4, 117.7, 116.7 and 116.7 (J = 2.6 Hz), 114.7, 110.8
and 110.6 (J = 20.9 Hz), 107.9 nd 107.6 (J = 26.3 Hz), 21.3
3 i
1H NMR (300 MHz, DMSO) δ 11.69 (bs, 1 H, OH), 10.57 (s, 1 H, NH), 7.98–7.83 (m, 3 H, H6, H3, H5),
7.74–7.68 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.83–6.76 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 2.29 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.1, 158.7, 144.8, 142.2, 129.4, 126.2 (q, J = 3.8 Hz), 124.6 (q, J =
271.4 Hz), 124.1 (q, J = 32.1 Hz), 120.8, 120.4, 117.7, 114.8, 21.3
3 k
1H NMR (300 MHz, DMSO) δ 11.60 (bs, 1 H, OH), 10.68 (s, 1 H, NH), 8.75 (t, 1 H, J = 2.19 Hz, H2Ј), 8.07
(ddd, 1 H, J = 8.24 Hz, J = 2.19 Hz, J = 0.83 Hz, H6Ј), 7.97 (ddd, 1 H, J = 8.24 Hz, J = 2.19 Hz, J = 0.83 Hz,
H4Ј), 7.86 (d, 1 H, J = 8.52 Hz, H6), 7.65 (t, 1 H, J = 8.24 Hz, H5Ј), 6.84–6.77 (m, 2 H, H3, H5), 2.30 (s,
3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.3, 158.8, 148.1, 144.9, 139.7, 130.3, 129.3, 126.9, 120.4, 118.7,
117.7, 115.0, 114.7, 21.4
3 l
1H NMR (300 MHz, DMSO) δ 12.36 (bs, 1 H, OH), 10.13 (s, 1 H, NH), 7.91 (d, 1 H, J = 8.79 Hz, H6),
7.52–7.43 (m, AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 6.80–6.67 (m, 4 H, H3, H5, H3Ј, H5Ј), 2.28 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.2, 160.0, 147.9, 144.4, 128.3, 127.5, 123.0, 120.0, 117.8, 113.6,
112.6, 40.6, 21.3
3 m
1H NMR (300 MHz, DMSO) δ 12.14 (bs, 1 H, OH), 10.24 (s, 1 H, NH), 7.89 (d, 1 H, J = 8.51 Hz, H6),
7.62–7.53 (m, AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 6.98–6.89 (m, AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.80–6.72 (m, 2 H, H3,
H5), 3.74 (s, 3 H, OCH3), 2.28 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.2, 159.8, 156.2, 144.6, 131.1, 128.6, 123.1, 120.1, 117.8, 114.0,
113.8, 55.4, 21.3
3 n
1H NMR (300 MHz, DMSO) δ 11.71 (bs, 1 H, OH), 10.56 (s, 1 H, NH), 7.99–7.92 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.91–7.83 (m, 3 H, H6, H3, H5), 6.83–6.75 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 4.28 (q, 2 H, J = 7.14 Hz,
CH2), 2.29 (s, 3 H, CH3), 1.30 (t, 3 H, J = 7.14 Hz, CH3)
13C NMR (75 MHz, DMSO) δ166.9, 165.5, 158.7, 144.8, 142.9, 130.3, 129.5, 125.1, 120.5, 120.2, 117.7,
114.8, 60.7, 21.3, 14.4

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 61
Table 2. continued.
Compound
NMR, δ
3 o
1H NMR (300 MHz, DMSO) δ 11.56 (bs, 1 H, OH), 10.6 (s, 1 H, NH), 7.95–7.88 (m, AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.85–7.77 (m, 3 H, H6, H3, H5), 6.83–6.76 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 2.29 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ166.9, 158.4, 144.8, 142.9, 133.4, 129.6, 120.8, 120.5, 119.2, 117.6, 115.1,
105.8, 21.3
4 a
1H NMR (300 MHz, DMSO) δ 12.43 (bs, 1 H, OH), 10.23 (bs, 1 H NH), 7.99 (d, 1 H, J = 8.79 Hz, H6),
7.72–7.63 (m, 2 H, H2Ј, H6Ј), 7.42–7.30 (m, 2 H, H3Ј, H5Ј), 7.17–7.08 (m, 1 H, H4Ј), 6.55 (dd, 1 H, J =
8.79 Hz, J = 2.74 HZ, H5), 6.49 (d, 1 H, J = 2.74 Hz, H3), 3.79 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.5, 164.0, 162.0, 138.3, 130.3, 128.9, 124.4, 121.5, 109.1, 106.5,
101.5, 55.6
4 b
4 c
4 d
1H NMR (300 MHz, DMSO) δ 12.54 (bs, 1 H, NH), 10.17 (bs, 1 H, OH), 7.99 (d, 1 H, J = 8.79 Hz, H6),
7.60–7.52 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.20–7.11 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.54 (dd, 1 H, J = 8.79 Hz,
J = 2.47 Hz, H5), 6.49 (d, 1 H, J = 2.48 Hz, H3), 3.78 (s, 3 H, OCH3), 2.27 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 167.5, 164.0, 162.2, 135.7, 133.5, 130.1, 129.3, 121.6, 109.0, 106.5,
101.5, 55.6, 20.7
1H NMR (300 MHz, DMSO) δ 12.29 (bs, 1 H, NH), 10.30 (bs, 1 H, OH), 7.96 (d, 1 H, J = 8.79 Hz, H6),
7.76-7.68 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.45-7.37 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.55 (dd, 1 H, J = 8.79 Hz,
J = 2.19 Hz, H5), 6.49 (d, 1 H, J = 2.19 Hz, H3), 3.78 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.4, 164.1, 161.9, 137.4, 130.4, 128.8, 128.0, 122.9, 109.1, 106.6,
101.5, 55.7
1H NMR (300 MHz, DMSO) δ 12.20 (bs, 1 H, NH), 10.33 (bs, 1 H, OH), 7.95 (d, 1 H, J = 8.79 Hz, H6), 7.89
(t, 1 H, J = 1.92 Hz, H2Ј), 7.60 (ddd, 1 H, J = 8.24 Hz, J = 1.92 Hz, J = 0.82 Hz, H6Ј), 7.37 (t, 1 H, J =
8.24 Hz, H5Ј), 7.17 (ddd, 1 H, J = 8.24 Hz, J = 1.92 Hz, J = 0.82 Hz, H4Ј), 6.56 (dd, 1 H, J = 8.79 Hz, J =
2.47 Hz, H5), 6.50 (d, 1 H, J = 2.47 Hz, H3), 3.78 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.4, 164.1, 161.7, 139.9, 133.2, 130.6, 130.5, 123.9, 120.7, 119.6,
109.3, 106.7, 101.5, 55.7
4 e
4 f
1H NMR (300 MHz, DMSO) δ 12.09 (bs, 1 H, NH), 10.36 (bs, 1 H, OH), 8.07 (d, 1 H, J = 2.47 Hz, H2Ј),
7.92 (d, 1 H, J = 8.79 Hz, H6), 7.66 (dd, 1 H, J = 8.79 Hz, J = 2.47 Hz, H5Ј), 7.58 (d, 1 H, J = 8.79 Hz, H6Ј),
6.55 (dd, 1 H, J = 8.79 Hz, J = 2.47 Hz, H5), 6.49 (d, 1 H, J = 2.47 Hz, H3), 3.78 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.4, 164.2, 161.6, 138.6, 131.1, 130.8, 130.6, 125.7, 122.3, 121.1,
109.3, 106.7, 101.5, 55.7
1H NMR (300 MHz, DMSO) δ 12.27 (bs, 1 H, NH), 10.30 (bs, 1 H, OH), 7.95 (d, 1 H, J = 8.79 Hz, H6),
7.70–7.63 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.57–7.50 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.55 (dd, 1 H, J = 8.79 Hz,
J = 2.47 Hz, H5), 6.49 (d, 1 H, J = 2.47 Hz, H3), 3.78 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.4, 164.1, 161.8, 137.8, 131.7, 130.4, 123.2, 116.1, 109.2, 106.6,
101.5, 55.7
4 g
4 h
1H NMR (300 MHz, DMSO) δ 12.41 (bs, 1 H, NH), 10.26 (bs, 1 H, OH), 7.96 (d, 1 H, J = 8.79 Hz, H6),
7.73–7.64 (m, 2 H, H2Ј, H6Ј), 7.25–7.14 (m, 2 H, H3Ј, H5Ј), 6.55 (dd, 1 H, J = 8.79 Hz, J = 2.47 Hz, H5),
6.49 (d, 1 H, J = 2.47 Hz, H3), 3.78 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.5, 164.1, 162.1, 160.5 and 157.3 (J = 240.8 Hz), 134.6 and 134.6 (J =
2.6 Hz), 130.2, 123.5 and 123.4 (J = 8.0 Hz), 115.7 and 115.4 (J = 22.3 Hz), 108.9, 106.6, 101.5, 55.7
1H NMR (300 MHz, DMSO) δ 12.20 (bs, 1 H, OH), 10.35 (bs, 1 H, NH), 7.96 (d, 1 H, J = 8.79 Hz, H6), 7.68
(dt (J = 11.54 Hz, J = 2.20 Hz, H6Ј), 7.50–7.33 (m, 2 H, H2Ј, H5Ј), 6.99–6.90 (m, 1 H, H4Ј), 6.56 (dd, 1 H,
J = 8.79, J = 2.47 Hz, H5), 6.50 (d, 1 H, J = 2.47 Hz, H3), 3.79 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.3, 164.1, 163.9 and 160.7 (J = 241.4 Hz), 161.6, 140.3 and 140.1 (J =
11.2 Hz), 130.6 and 130.4 (J = 11.5 Hz), 116.9 and 116.9 (J = 2.6 Hz), 110.8 and 110.5 (J = 21.2 Hz),
109.4, 108.1 and 107.8 (J = 26.3 Hz), 106.6, 101.5, 55.7

62 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Table 2. continued.
Compound
NMR, δ
4 i
1H NMR (300 MHz, DMSO) δ 12.14 (bs, 1 H, OH), 10.48 (s, 1 H, NH), 8.01–7.88 (m, 3 H, H6, H3Ј, H5Ј),
7.78–7.64 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 6.57 (dd, 1 H, J = 9.06 Hz, J = 2.47 Hz, H6), 6.51 (d, 1 H, J = 2.47
Hz, H8), 3.79 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.4, 164.2, 161.5, 142.2, 130.8, 126.2 (q, J = 3.8 Hz), 124.6 (q, J = 271.4
Hz), 124.1 (q, J = 32.4 Hz), 120.9, 109.5, 106.7, 101.5, 55.7
4 j
1H NMR (300 MHz, DMSO) δ 10.66 (bs, 1 H, NH), 8.27–8.20 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 8.01–7.91 (m,
3 H, H6, H2Ј, H6Ј), 6.60–6.55 (m, 1 H, H5), 6.53–6.50 (m, 1 H, H3), 3.79 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.2, 164.3, 161.2, 144.9, 142.7, 131.1, 125.0, 120.4, 109.8, 106.8,
101.5, 55.7
4 k
1H NMR (300 MHz, DMSO) δ 12.10 (bs, 1 H, OH), 10.55 (bs, 1 H, NH), 8.71 (t, 1 H, J = 2.19 Hz, H2Ј), 8.07
(ddd, 1 H, J = 8.24 Hz, J = 2.20 Hz, J = 0.82 Hz, H6Ј), 7.99–7.92 (m, 2 H, H6, H4Ј), 7.63 (t, 1 H, J =
8.25 Hz, H5Ј), 6.57 (dd, 1 H, J = 8.79 Hz, J = 2.47 Hz, H5), 6.50 (d, 1 H, J = 2.47 Hz, H3), 3.79 (s, 3 H,
OCH3)
13C NMR (75 MHz, DMSO) δ 167.6, 164.3, 161.7, 148.1, 139.7, 130.6, 130.3, 127.0, 118.6, 115.2,
109.2, 106.7, 101.5, 55.7
4 m
4 n
1H NMR (300 MHz, DMSO) δ 12.63 (bs, 1 H, OH), 10.14 (bs, 1 H, NH), 7.97 (d, 1 H, J = 8.79 Hz, H6),
7.59–7.52 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 6.97–6.89 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 6.53 (dd, 1 H, J = 8.79 Hz,
J = 2.47 Hz, H5), 6.47 (d, 1 H, J = 2.47 Hz, H3), 3.78 (s, 3 H, OCH3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 167.6, 164.0, 162.4, 156.2, 131.1, 129.9, 123.4, 114.0, 108.8, 106.5,
101.5, 55.6, 55.4
1H NMR (300 MHz, DMSO) δ 12.15 (bs, 1 H, OH), 10.46 (s, 1 H, NH), 7.98 (d, 1 H, J = 8.79 Hz, H5),
7.97–7.92 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.89–7.82 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 6.57 (dd, 1 H, J = 8.79 Hz,
J = 2.47 Hz, H6), 6.51 (d, 1 H, J = 2.47 Hz, H8), 4.28 (q, 2 H, J = 7.10 Hz, CH2), 3.79 (s, 3 H, OCH3), 1.30
(t, 3 H, J = 7.10 Hz, CH3)
13C NMR (75 MHz, DMSO) δ167.3, 165.5, 164.2, 161.4, 142.9, 130.8, 130.3, 125, 120.3, 109.6, 106.7,
101.5, 60.7, 55.7, 14.4
4 o
5 i
1H NMR (300 MHz, DMSO) δ 12.01 (bs, 1 H, OH), 10.5 (s, 1 H, NH), 7.94 (d, 1 H, J = 9.07, H5), 7.93–7.87
(m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.84–7.76 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 6.57 (dd, 1 H, J = 9.07 Hz, J = 2.47 Hz,
H6), 6.51 (d, 1 H, J = 2.47 Hz, H8), 3.79 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ167.2, 164.2, 161.2, 142.9, 133.4, 131, 120.9, 119.3, 109.7, 106.8, 105.8,
101.5, 55.7
1H NMR (300 MHz, DMSO) δ 11.58 (bs, 1 H, OH), 10.59 (s, 1 H, NH), 7.95 (d, 1 H, J = 2.47 Hz, H6),
7.92–7.85 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.71–7.65 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.53 (dd, 1 H, J = 8.79 Hz,
J = 2.47 Hz, H4), 6.93 (d, 1 H, J = 8.79 Hz, H3)
13C NMR (75 MHz, DMSO) δ 165.2, 156.9, 142.0, 136.1, 131.7, 126.3 (q, J = 3.7 Hz), 124.5 (q, J = 271.7
Hz), 124.3 (q, 32.1 Hz), 121.1, 120.7, 119.6, 110.4
5 n
1H NMR (300 MHz, DMSO) δ 11.65 (bs, 1 H, OH), 10.59 (s, 1 H, NH), 8.00 (d, 1 H, J = 2.47 Hz, H6),
7.98–7.92 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.71–7.65 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.65 (dd, 1 H, J = 8.79 Hz,
J = 2.48 Hz, H4), 6.96 (d, 1 H, J = 8.79 Hz, H3), 4.28 (q, 2 H, J = 7.14 Hz, CH2), 1.30 (t, 3 H, J = 7.14 Hz,
CH3)
13C NMR (75 MHz, DMSO) δ 165.5, 165.1, 156.9, 142.8, 136.1, 131.7, 130.4, 125.2, 121.1, 120.0,
119.6, 110.5, 60.7, 14.4
5 o
1H NMR (300 MHz, DMSO) δ 11.52 (bs, 1 H, OH), 10.66 (s, 1 H, NH), 7.94 (d, 1 H, J = 2.47 Hz, H6),
7.93–7.87 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.84–7.79 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.56 (dd, 1 H, J = 8.79 Hz,
J = 2.48 Hz, H4), 6.96 (d, 1 H, J = 8.79 Hz, H3)
13C NMR (75 MHz, DMSO) δ 165.1, 156.6, 142.8, 136.0, 133.4, 131.8, 121.4, 120.6, 119.6, 119.2,
110.4, 106.0

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 63
Table 2. continued.
Compound
NMR, δ
6 i
1H NMR (300 MHz, DMSO) δ 10.66 (bs, 1 H, NH), 7.97–7.89 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.88 (d, 1 H, J =
2.74 Hz, H6), 7.75–7.68 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.46 (dd, 1 H, J = 8.79 Hz, J = 2.74 Hz, H4), 7.02 (d,
1 H, J = 8.79 Hz, H3)
13C NMR (75 MHz, DMSO) δ 165.3, 156.5, 142.1, 133.3, 128.8, 126.3, 126.2, 124.5 (q, J = 271.4 Hz),
124.3 (q, J = 32.1 Hz), 123.0, 120.6 (d, J = 6.9 Hz), 120.6
6 n
1H NMR (300 MHz, DMSO) δ 10.65 (bs, 1 H, NH), 7.98–7.92 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.88 (d over-
lapped, 1 H, J = 2.75 Hz, H6), 7.88–7.83 (m AAЈ, BBЈ overlapped 2 H, H2Ј, H6Ј), 7.45 (dd, 1 H, J =
8.79 Hz, J = 2.74 Hz, H4), 8.05 (d, 1 H, J = 8.79 Hz, H3), 4.29 (q, 2 H, J = 7.14 Hz, CH2), 1.31 (t, 3 H, J =
7.15 Hz, CH3)
13C NMR (75 MHz, DMSO) δ 165.5, 165.1, 156.5, 142.8, 133.2, 130.4, 128.9, 125.2, 123.0, 120.6,
120.0, 119.2, 60.7, 14.4
6 o
7 i
1H NMR (300 MHz, DMSO) δ 11.49 (bs, 1 H, OH), 10.67 (bs, 1 H, NH), 7.94–7.88 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.85–7.79 (m AAЈ, BBЈoverlapped, 2 H, H2Ј, H6Ј), 7.82 (d overlapped, 1 H, J = 2.47 Hz, H6), 7.46
(dd, 1 H, J = 8.79 Hz, J = 2.47 Hz, H4), 7.02 (d, 1 H, J = 8.79 Hz, H3)
13C NMR (75 MHz, DMSO) δ 165.2, 156.3, 142.8, 133.5, 133.2, 128.9, 123.0, 120.9, 120.6, 119.2,
119.1, 106.0
1H NMR (300 MHz, DMSO) δ 11.41 (bs, 1 H, OH), 10.66 (bs, 1 H, NH), 7.98–7.89 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.75–7.70 (m AAЈ, BBЈ overlapped, 2 H, H2Ј, H6Ј), 7.68 (dd overlapped, 1 H, J = 9.20 Hz, J = 3.02
Hz, H6), 7.36–7.26 (m, 1 H, H4), 7.01 (dd, 1 H, J = 9.20 Hz, J = 4.67 Hz, H3)
13C NMR (75 MHz, DMSO) δ 165.3 (d, J = 2.3 Hz), 155.1 (d, J = 235.4 Hz), 154.0, 142.1, 126.3 (q, J = 4.0
Hz), 124.5 (q, J = 271.4 Hz), 124.3 (q, J = 32.3 Hz), 120.7 (d, J = 23.2 Hz), 120.6, 119.4 (d, J = 6.7 Hz),
118.7 (d, J = 7.7 Hz), 115.3 (d, J = 24.6 Hz)
7 n
1H NMR (300 MHz, DMSO) δ 11.43 (bs, 1 H, OH), 10.64 (bs, 1 H, NH), 8.00–7.91 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.90–7.82 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.69 (dd, 1 H, J = 9.28 Hz, J = 3.16 Hz, H6), 7.35–7.25 (m,
1 H, H4), 7.01 (dd, 1 H, J = 9.28 Hz, J = 4.53 Hz, H3), 4.29 (q, 2 H, J = 7.14 Hz, CH2), 1.31 (t, 3 H, J = 7.15
Hz, CH3)
13C NMR (75 MHz, DMSO) δ 165.5, 165.2 (d, J = 2.3 Hz), 155.2 (d, J = 235.4 Hz), 154.0, 142.8, 130.5 (d,
J = 12.9 Hz), 125.2, 120.7 (d, J = 23.5 Hz), 120.0, 119.5, 118.7 (d, J = 7.5 Hz), 115.3 (d, J = 24.3 Hz),
60.7, 14.4
7 o
1H NMR (300 MHz, DMSO) δ 11.32 (bs, 1 H, OH), 10.69 (bs, 1 H, NH), 7.95–7.88 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.85–7.79 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.64 (dd, 1 H, J = 9.61 Hz, J = 3.29 Hz, H6), 7.35–7.25 (m,
1 H, H4), 7.02 (dd, 1 H, J = 9.07 Hz, J = 4.67 Hz, H3)
13C NMR (75 MHz, DMSO) δ 165.2 (d, J = 2.0 Hz), 155.2 (d, J = 235.4 Hz), 153.7 (d, J = 1.7 Hz), 142.7,
133.5, 120.7 (d, J = 23.2 Hz), 120.6, 119.8 (d, J = 6.9 Hz), 119.2, 118.7 (d, J = 7.7 Hz), 115.4 (d, J = 24.3
Hz), 106.0
8 g
8 h
1H NMR (300 MHz, DMSO) δ 11. 56 (bs, 1 H, OH), 10.39 (bs, 1 H, NH), 7.76 (d, 1 H, J = 1.65 Hz, H6),
7.74–7.67 (m, 2 H, H2Ј, H6Ј), 7.27–7.14 (m, 3 H, H4, H3Ј, H5Ј), 6.87 (d, 1 H, J = 8.52 Hz, H3), 2.27 (s,
3 H, CH3)
13C NMR (75 MHz, DMSO) δ 166.9, 158.8 (d, J = 240.8 Hz), 156.7, 134.7 (d, J = 2.6 Hz), 134.6, 129.0,
127.9, 123.1 (d, J = 8.1 Hz), 117.3, 117.0, 115.6 (d, J = 22.3 Hz), 20.3
1H NMR (300 MHz, DMSO) δ 11.37 (bs, 1 H, OH), 10.48 (bs, 1 H, NH), 7.77–7.68 (m, 2 H, H2Ј, H6Ј),
7.50–7.33 (m, 2 H, H6, H5Ј), 7.27–7.21 (m, 1 H, H4Ј), 6.99–6.91 (m, 1 H, H4), 6.88 (d, 1 H, J = 8.24 Hz,
H3), 2.26 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 166.7, 162.3 (d, J = 241.4 Hz), 156.1, 140.3 (d, J = 11.2 Hz), 134.6, 130.5
(d, J = 9.4 Hz), 129.3, 128.0, 117.6, 117.3, 116.6 (d, J = 2.9 Hz), 110.7, (d, J = 20.9), 107.7 (d, J =
26.3 Hz), 20.3

64 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Table 2. continued.
Compound
NMR, δ
8 i
1H NMR (300 MHz, DMSO) δ 11.32 (bs, 1 H, OH), 10.61 (s, 1 H, NH), 7.98 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј),
7.77–7.66 (m, 3 H, H6, H2Ј, H6Ј), 7.24 (dd, 1 H, J = 8.24 Hz, J = 1.65 Hz, H4), 6.89 (d, 1 H, J = 8.24 Hz,
H3), 2.27 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 166.8, 155.9, 142.3, 134.6, 129.5, 128.1, 126.2 (d, J = 3.7 Hz), 124.6 (q,
271.4 Hz), 124.1 (q, J = 32.4 Hz), 120.7, 117.8, 117.2, 20.2
8 n
8 o
9 a
9 b
9 c
9 d
9 e
9 f
1H NMR (300 MHz, DMSO) δ 10.60 (d, 1 H, J = 5.22 Hz, NH), 8.01–7.91 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј),
7.91–7.81 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.73 (s, 1 H, H6), 7.24 (dd, 1 H, J = 8.24 Hz, J = 1.93 Hz, H4), 6.90
(d, 1 H, J = 8.24 Hz, H3), 4.28 (q, 2 H, J = 7.14 Hz, CH2), 2.26 (s, 3 H, CH3), 1.30 (t, 3 H, J = 7.14 Hz, CH3)
13C NMR (75 MHz, DMSO) δ 167.0, 165.5, 156.0, 143.0, 134.7, 130.4, 129.5, 128.1, 125.1, 120.1,
117.8, 117.3, 60.8, 20.3, 14.5
1H NMR (300 MHz, DMSO) δ 11.20 (bs, 1 H, OH), 10.64 (bs, 1 H, NH), 7.95–7.89 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.84–7.78 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.68 (d, 1 H, J = 2.20 Hz, H6), 7.24 (dd, 1 H, J = 8.38 Hz, J =
2.20 Hz, H7), 6.90 (d, 1 H, J = 8.38 Hz, H8), 2.26 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 166.7, 155.6, 143.0, 134.6, 133.4, 129.6, 128.1, 120.6, 119.2, 118.2,
117.2, 105.8, 20.2
1H NMR (300 MHz, DMSO) δ 11.36 (bs, 1 H, OH), 10.42 (bs, 1 H, NH), 7.73–7.65 (m, 2 H, H2Ј, H6Ј), 7.50
(d, 1 H, J = 3.02 Hz, H6), 7.41–7.33 (m, 2 H, H3Ј, H5Ј), 7.19–7.09 (m, 1 H, H4Ј), 7.06 (dd, 1 H, J = 8.93 Hz,
J = 3.02 Hz, H4), 6.92 (d, 1 H, J = 8.93 Hz, H3), 3.75 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.2, 152.4, 152.0, 138.3, 129.0, 124.4, 121.2, 120.8, 118.4, 117.7,
112.8, 55.9
1H NMR (300 MHz, DMSO) δ 11.44 (bs, 1 H, OH), 10.35 (bs, 1 H, NH), 7.59–7.53 (m AAЈ, BBЈ, 2 H, H2Ј,
H6Ј), 7.50 (d, 1 H, J = 3.30 Hz, H6), 7.20–7.14 (m AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.05 (dd, 1 H, J = 8.93 Hz, J =
3.30 Hz, H4), 6.91 (d, 1 H, J = 8.93 Hz, H3), 3.75 (s, 3 H, OCH3), 2.28 (s, 3 H, CH3)
13C NMR (75 MHz, DMSO) δ 166.2, 152.7, 152.0, 135.7, 133.5, 129.4, 121.3, 120.8, 118.4, 117.5,
112.7, 55.9, 20.7
1H NMR (300 MHz, DMSO) δ 11.21 (bs, 1 H, OH), 10.47 (bs, 1 H, NH), 7.77–7.69 (m AAЈ, BBЈ, 2 H, H2Ј,
H6Ј), 7.45 (d, 1 H, J = 3.02 Hz, H6), 7.44–7.37 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 7.06 (dd, 1 H, J = 8.79 Hz, J =
3.02 Hz, H4), 6.92 (d, 1 H, J = 8.80 Hz, H3), 3.75 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.2, 152.2, 152.1, 137.4, 128.9, 128.0, 122.6, 120.8, 118.3, 117.9,
112.9, 55.9
1H NMR (300 MHz, DMSO) δ 11.14 (bs, 1 H, OH), 10.50 (bs, 1 H, NH), 7.92 (t, 1 H, J = 1.92 Hz, H2Ј),
7.62–7.57 (m, 1 H, H6Ј), 7.44 (d, 1 H, J = 3.02 Hz, H6), 7.39 (t, 1 H, J = 8.24 Hz, H5Ј), 7.21–7.16 (m, 1 H,
H4Ј), 7.06 (dd, 1 H, J = 8.93 Hz, J = 3.02 Hz, H4), 6.93 (d, 1 H, J = 8.93 Hz, H3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.2, 152.1, 152.0, 139.9, 133.3, 130.6, 124.0, 120.8, 120.4, 119.3,
118.3, 118.1, 113.0, 55.9
1H NMR (300 MHz, DMSO) δ 11.06 (bs, 1H, OH), 10.56 (bs, 1 H, NH), 8.11 (d, 1 H, J = 2.20 Hz, H2Ј), 7.66
(dd, 1 H, J = 8.79 Hz, J = 2.20 Hz, H6Ј), 7.60 (d, 1 H, J = 8.79 Hz, H5Ј), 7.41 (d, 1 H, J = 3.02 Hz, H6), 7.06
(dd, 1 H, J = 8.93 Hz, J = 3.02 Hz, H4), 6.93 (d, 1 H, J = 8.93 Hz, H3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.2, 152.1, 151.9, 138.6, 131.2, 130.8, 125.7, 122.1, 120.9, 120.8,
118.3, 118.2, 113.0, 55.9
1H NMR (300 MHz, DMSO) δ 10.48 (bs, 1 H, NH), 7.72–7.63 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.59–7.50 (m
AAЈ, BBЈ, 2 H, H3Ј, H5Ј), 7.44 (d, 1 H, J = 3.02 Hz, H6), 7.05 (dd, 1 H, J = 8.79 Hz, J = 3.02 Hz, H4), 6.92
(d, 1 H, J = 8.79 Hz, H3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.1, 152.2, 152.1, 137.8, 131.8, 123.0, 120.8, 118.4, 118.0, 116.1,
112.9, 55.9
9 g
1H NMR (300 MHz, DMSO) δ 11.33 (bs, 1 H, OH), 10.43 (bs, 1 H, NH), 7.77 (m, 2 H, H2Ј, H6Ј), 7.48 (d,
1 H, J = 3.02 Hz, H6), 7.27–7.15 (m, 2 H, H3Ј, H5Ј), 7.06 (dd, 1 H, J = 8.93 Hz, J = 3.02 Hz, H4), 6.91 (d,
1 H, J = 3.02 Hz, H3), 3.75 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.3, 158.9 (d, J = 248.8 Hz), 152.5, 152.0, 134.7 (d, J = 2.6 Hz), 123.2 (d,
J = 8.0 Hz), 120.8, 118.4, 117.5 115.6 (d, J = 22.3 Hz), 112.7, 55.9

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 65
Table 2. continued.
Compound
NMR, δ
9 h
9 i
1H NMR (300 MHz, DMSO) δ 10.56 (bs, 1 H, NH), 7.77–7.67 (m, 1 H, H6Ј), 7.50–7.32 (m, H6, H2Ј, H4Ј,
H5Ј), 7.06 (dd, 1 H, J = 9.07 Hz, J = 3.02 Hz, H4), 6.99–6.90 (m, 1 H, H3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.1, 162.3 (d, J = 241.4 Hz), 152.1, 152.0, 140.2 (d, J = 10.9 Hz), 130.5
(d, J = 9.5 Hz), 120.7, 118.4, 118.2, 116.6 (d, J = 2.6 Hz), 113.0, 110.7 (d, J = 20.9 Hz), 107.7 (d, J =
26.0 Hz), 55.8
1H NMR (300 MHz, DMSO) δ 11.12 (bs, 1 H, OH), 10.67 (bs, 1 H, NH), 7.97–7.90 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.77–7.68 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.45 (d, 1 H, J = 3.02 Hz, H6), 7.07 (dd, 1 H, J = 9.07 Hz, J =
3.02 Hz, H4), 6.94 (d, 1 H, J = 9.07 Hz, H3), 3.75 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.2, 159.1, 151.9, 142.2, 126.2 (q, J = 3.7 Hz), 124.6 (q, J = 271.4 Hz),
124.1 (q, J = 32.1 Hz), 120.8, 120.7, 118.4, 113.2, 55.8
9 j
1H NMR (300 MHz, DMSO) δ 10.99 (bs, 1 H, OH), 10.84 (bs, 1 H, NH), 8.30–8.20 (m, AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 8.03–7.93 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.39 (d, 1 H, J = 3.02 Hz, H6), 7.07 (dd, 1 H, J = 9.06 Hz, J =
3.02 Hz, H4), 6.95 (d, 1 H, J = 9.07 Hz, H3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.1, 152.2, 151.4, 144.9, 142.8, 125.1, 120.8, 120.3, 118.9, 118.3,
113.3, 55.8
9 k
9 m
9 n
1H NMR (300 MHz, DMSO) δ 11.06 (bs, 1 H, OH), 10.72 (bs, 1 H, NH), 8.76 (t, 1 H, J = 1.92 Hz, H2Ј),
8.09–8.02 (m, 2 H, H4Ј, H6Ј), 7.65 (t, 1 H, J = 8.24 Hz, H5Ј), 7.44 (d, 1 H, J = 3.02 Hz, H6), 7.07 (dd, 1 H,
J = 8.79 Hz, J = 3.02 Hz, H4), 6.94 (d, 1 H, J = 8.79 Hz, H3), 3.75 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.5, 152.1, 152.0, 148.1, 139.7, 130.3, 126.9, 120.9, 118.7, 118.4,
118.1, 114.9, 113.0, 55.9
1H NMR (300 MHz, DMSO) δ 11.54 (bs, 1 H, OH), 10.32 (bs, 1 H, NH), 7.61–7.54 (m AAЈ, BBЈ, 2 H, H2Ј,
H6Ј), 7.51 (d, 1 H, J = 3.02 Hz, H6), 7.05 (dd, 1 H, J = 8.79 Hz, J = 3.02 Hz, H4), 6.98–6.92 (m AAЈ, BBЈ,
2 H, H3Ј, H5Ј), 6.90 (d, 1 H, J = 8.79 Hz, H3), 3.75 (s, 3 H, OCH3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.4, 156.3, 153.0, 151.9, 131.1, 123.1, 120.8, 118.4, 117.1, 114.1,
112.4, 55.9, 55.4
1H NMR (300 MHz, DMSO) δ 11.13 (bs, 1 H, OH), 10.65 (bs, 1 H, NH), 7.99–7.93 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.89–7.83 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.45 (d, 1 H, J = 3.16 Hz, H6), 7.06 (dd, 1 H, J = 9.06 Hz, J =
3.16 Hz, H4), 6.94 (d, 1 H, J = 9.07 Hz, H3), 4.29 (q, 2 H, J = 7.14 Hz, CH2), 3.75 (s, 3 H, OCH3), 1.31 (t,
3 H, J = 7.14 Hz, CH3)
13C NMR (75 MHz, DMSO) δ 166.1, 165.5, 152.2, 151.8, 142.9, 130.4, 125.1, 120.8, 120.1, 118.4,
118.3, 113.2, 60.7, 55.8, 14.4
9 o
1H NMR (300 MHz, DMSO) δ 11.01 (bs, 1 H, OH), 10.69 (bs, 1 H, NH), 7.96–7.88 (m AAЈ, BBЈ, 2 H, H3Ј,
H5Ј), 7.86–7.77 (m AAЈ, BBЈ, 2 H, H2Ј, H6Ј), 7.40 (d, 1 H, J = 3.02 Hz, H6), 7.06 (dd, 1 H, J = 8.92 Hz, J =
3.02 Hz, H4), 6.94 (d, 1 H, J = 8.92 Hz, H3), 3.74 (s, 3 H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.1, 152.2, 151.5, 142.9, 133.5, 120.8, 120.7, 119.2, 118.7, 118.3,
113.3, 105.9, 55.8
Table 3. Minimum inhibitory concentrations of salicylanilides substituted in position 4 and 5.
MIC (µmol/L)
Incubation 14 d/21 d
Compounds
R1
R2
M. tuberculosis
M. kansasii
M. avium
1 ac
1 bc
1 cc
1 dc
1 ec
H
H
H
H
H
H
4-CH3
4-Cl
3-Cl
3,4-Cl2
62.5/62.5
62.5/62.5
31/31
16/16
8/8
125/250
62.5/125
31/31
8/8
62.5/125
31/62.5
31/31
31/31
16/31
4/8

66 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Table 3. continued.
MIC (µmol/L)
Incubation 14 d/21 d
Compounds
R1
R2
M. tuberculosis
M. kansasii
M. avium
1 fc
1 gc
1 hc
1 id
1 jc
1 kc
1 lc
1 mc
1 nd
1 od
1 pd
2 a
2 b
2 c
2 d
2 e
2 f
2 g
2 h
2 i
2 k
2 l
2 m
2 n
2 o
3 a
3 b
3 c
3 d
3 e
3 f
3 g
3 h
3 i
3 k
3 l
3 m
3 n
3 o
4 a
4 b
4 c
4 d
4 e
4 f
H
H
H
H
H
H
H
H
H
H
H
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Cl
4-Br
4-F
3-F
16/31
62.5/62.5
31/31
8/8
16/31
62.5/125
62.5/62.5
16/16
16/16
62.5/a
250/a
250/250
32/62.5
62.5/125
32/62.5
8/16
31/31
31/62.5
62.5/62.5
16/32
31/31
31/a
125/250
62.5/125
32/32
32/62.5
62.5/62.5
16/16
16/16
8/8
16/16
16/16
16/16
32/32
32/32
8/8
16/16
16/16
16/16
16/16
16/32
a/a
4-CF3
4-NO2
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
4-CSNH2
H
4-CH3
4-Cl
3-Cl
3,4-Cl2
4-Br
4-F
8/16
16/16
250/250
62.5/62.5
16/32
32/32
32/32
8/16
16/31
4/4
4/4
4/4
4/4
16/16
8/16
4/4
8/8
a/a
8/16
4/8
4/8
8/8
4/4
16/32
32/32
4/4
16/16
a/a
16/31
8/8
16/16
a/a
a/a
a/a
a/a
8/16
a/a
a/a
62.5/62.5
a/a
31/31
a/a
a/a
a/a
16/32
a/a
a/a
3-F
4-CF3
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
H
4-CH3
4-Cl
3-Cl
8/16
8/8
8/8
31/62
250/a
62/a
a/a
4-Cl
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-CH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
a/a
a/a
16/a
8/8
a/a
a/a
32/62.5
a/a
31/62
a/a
3,4-Cl2
4-Br
4-F
4/8
a/a
16/a
8/16
16/a
4/8
31/62
16/31
8/8
16/16
16/16
16/32
16/32
4/4
8/16
32/32
16/16
8/8
3-F
4-CF3
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
H
4-CH3
4-Cl
3-Cl
3,4-Cl2
4-Br
4-F
3-F
4-CF3
62/a
a/a
16/32
62.5/125
62.5/62,5
32/a
32/32
32/32
a/a
a/a
62.5/62.5
a/a
a/a
a/a
62.5/a
a/a
4 g
4 h
4 i
32/a
a/a
a/a
a/a

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 67
Table 3. continued.
MIC (µmol/L)
Incubation 14 d/21 d
Compounds
R1
R2
M. tuberculosis
M. kansasii
M. avium
4 j
4 k
4 m
4 n
4-OCH3
4-OCH3
4-OCH3
4-OCH3
4-OCH3
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Br
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-Cl
5-F
4-NO2
3-NO2
4-OCH3
4-COOEt
4-CN
H
4-CH3
4-Cl
3-Cl
8/8
4/4
8/8
32/32
62.5/62.5
a/a
16/16
32/32
a/a
a/a
a/a
31/62.5
a/a
16/16
31/31
8/8
16/16
31/31
31/31
1/1
31/31
a/a
a/a
a/a
8/8
16/31
16/16
8/8
8/16
16/16
8/8
4/8
a/a
a/a
4 o
16/16
16/31
16/16
4/4
4/8
4/4
4/4
8/8
8/8
1/1
5 ac
5 bc
5 cc
5 dc
5 ec
5 fc
5 gc
5 hc
5 i
31/31
16/a
8/16
16/16
8/8
8/8
8/8
8/16
2/4
16/16
a/a
a/a
8/8
8/8
4/4
4/4
8/8
4/8
4/4
4/4
4/4
8/8
1/1
4/4
8/8
8/8
4/8
16/32
16/16
31/62.5
31/31
8/8
a/a
4/4
4/8
8/8
3,4-Cl2
4-Br
4-F
3-F
4-CF3
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
H
4-CH3
4-Cl
3-Cl
5 kc
5 lc
5 mc
5 n
8/8
a/a
a/a
4/4
5 o
2/4
6 ac
6 bc
6 cc
6 dc
6 ec
6 fc
6 gc
6 hc
6 i
31/31
16/31
4/4
4/8
4/8
8/16
16/16
8/8
2/2
4/8
3,4-Cl2
4-Br
4-F
16/16
31/31
8/8
3-F
4-CF3
4-NO2
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
H
4-CH3
4-Cl
3-Cl
3,4-Cl2
4-Br
4-F
6 jc
8/8
6 kc
6 lc
8/8
a/a
a/a
8/8
16/16
31/62.5
31/31
32/32
32/32
31/62.5
16/31
16/16
16/16
8/8
6 mc
6 n
6 o
4/8
7 ac
7 bc
7 cc
7 dc
7 ec
7 fc
7 gc
7 hc
7 i
31/62.5
31/62.5
8/8
8/16
4/4
5-F
5-F
5-F
5-F
5-F
5-F
5-F
5-F
5-F
5-F
5-F
5-F
4/8
8/16
16/16
31/31
4/4
8/16
31/a
62.5/62.5
8/16
8/16
16/16
31/31
16/16
16/31
a/a
16/16
62.5/a
32/32
3-F
16/16
8/8
16/31
a/a
62.5/62.5
32/32
16/32
4-CF3
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
7 kc
7 lc
7 mc
7 n
7 o
5-F

68 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Table 3. continued.
MIC (µmol/L)
Incubation 14 d/21 d
Compounds
R1
R2
M. tuberculosis
M. kansasii
M. avium
8 a
8 b
8 c
8 d
8 e
8 f
8 g
8 h
8 i
8 j
8 k
8 l
8 m
8 n
8 o
9 a
9 b
9 c
9 d
9 e
9 f
9 g
9 h
9 i
9 j
9 k
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-CH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-OCH3
5-NO2
5-NO2
5-NO2
5-NO2
5-NO2
5-NO2
5-NO2
5-NO2
5-NO2
5-NO2
5-NO2
INH
H
4-CH3
4-Cl
3-Cl
3,4-Cl
4-Br
62.5/62.5
32/62.5
8/16
16/32
28/16
16/32
b/b
b/b
4/8
16/32
32/32
a/a
62.5/125
250/a
32/a
16/16
8/16
32/a
a/a
a/a
8/16
8/8
62.5/62.5
32/32
32/62.5
32/a
16/a
32/32
b/b
b/b
8/16
16/a
62.5/a
a/a
62.5/a
>1000/>1000
1000/1000
125/125
a/a
4-F
3-F
4-CF3
4-NO2
3-NO2
4-N(CH3)2
4-OCH3
4-COOEt
4-CN
H
4-CH3
4-Cl
3-Cl
3,4-Cl2
4-Br
4-F
a/a
a/a
a/a
125/a
>1000/>1000
500/1000
a/a
>1000/>1000
1000/1000
a/a
a/a
a/a
a/a
a/a
a/a
a/a
a/a
125/a
a/a
a/a
a/a
a/a
a/a
a/a
8/16
8/16
8/8
8/16
8/8
62.5/a
62.5/62.5
62.5/62.5
a/a
a/a
125/125
16/a
a/a
a/a
a/a
a/a
62.5/a
125/a
a/a
a/a
a/a
3-F
4-CF3
4-NO2
3-NO2
4-OCH3
4-COOEt
4-CN
H
4-CH3
4-Cl
3-Cl
a/a
a/a
a/a
a/a
9 m
9 n
9 o
a/a
a/a
a/a
a/a
10 ac
10 bc
10 cc
10 dc
10 ec
10 fc
10 gc
10 hc
10 kc
10 lc
10 mc
16/31
16/16
16/16
16/16
8/8
16/16
16/31
31/31
31/31
31/31
31/31
4/4
250/250
125/125
62.5/62.5
62.5/62.5
31/62.5
31/62.5
125/125
62.5/125
125/125
500/1000
250/250
500/500
3,4-Cl2
4-Br
4-F
8/8
8/8
3-F
16/16
8/16
8/16
8/16
500/500
3-NO2
4-N(CH3)2
4-OCH3
a
MIC could not be determined due to a low solubility.
not tested against the given strain.
data from ref. [7].
b
c
d
data from ref. [6].

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 69
Table 4. Regression coefficients of the indicator parameters In in equations (1)–(3).
Equation
M. tubeculosis.
M. kansasii
M. avium
21 d
21 d
21 d
(1)–(3)
(1)–(3)
(1)–(3)
(1)–(3)
(1)–(3)
(1)–(3)
(1)–(3)
(1)–(3)
(1)–(3)
I(5-Br)
–0.611
( 0.106)
–0.435
( 0.104)
–0.224
( 0.109)
–0.209
( 0.109)
0.212
–0.568
( 0.121)
–0.895
( 0.111)
–0.474
( 0.118)
–0.619
( 0.122)
–0.019
–0.226
( 0.120)
–0.396
( 0.108)
–0.261
( 0.113)
0.478
( 0.17)
0.221
( 0.142)
0.296
( 0.305)
–0.390
( 0.106)
–0.028
( 0.167)
0.048
I(5-Cl)
I(5-F)
I(5-NO2)
I(5-CH3)
I(5-OCH3)
I(4-Cl)
( 0.113)
0.607
( 0.147)
a
a
( 0.177)
–0.545
( 0.104)
–0.241
( 0.120)
–0.373
( 0.109)
–0.578
( 0.109)
–0.160
( 0.172)
0.019
I(4-CH3)
I(4-OCH3)
( 0.193)
( 0.142)
a
Evaluation hampered by the low solubility of the compounds.
anilides includes the results obtained after 21 days of
incubation, as those obtained after 14 days of incubation
were similar.Equations (1)–(3) are given without regres-
sion coefficients of the indicator parameters; these are
summarised in Table 4. The numbers of compounds in
the equations analysing structure-activity relationships
against various strains are different, as in a number of
cases the minimum inhibitory concentration could not be
determined due to the limited solubility of the com-
pounds.
Upon using the indicator parameters, the biological ac-
tivity can be predicted, but only within the framework of
the selection of substituents employed. Thus, we at-
tempted to find correlations with physical and chemical
parameters.A classical Hansch-type equation, using the
second power of the hydrophobic parameters, was not
statistically significant. Salicylanilides possess the hy-
droxy and amide functional groups.The influence of the
substituents in the acyl part of the molecule was thus ex-
pressed with the corresponding Hammett constant val-
ues with regard to both centers, and their hydrophobic
–
constants π (with regard to the hydroxy group) and mo-
log MICM..tuber.Ј21d = – 0.498 ( 0.078) σ
– 0.249 ( 0.050) π + In
lecular refraction (MR) values were used as additional
parameters. The results are summarised in equations
(4)–(6).
+ 1.713 ( 0.074)
(1)
r = 0.811
s = 0.278 n = 110
F = 17.1
log MICM..tuber. 21d = 0.577 ( 0.182) (σ1)CO
– 0.777 ( 0.194) (σ1)OH
log MICM..kans.21d = – 0.205 ( 0.100) σ
–
– 0.336 ( 0.058) π + In
+ 1.890 ( 0.087)
–
– 0.578 ( 0.098) π1
(2)
(3)
– 0.494 ( 0.087) σ2
– 0.246 ( 0.056) π2
+ 0.025 ( 0.014)RM
r = 0.797
s = 0.299
n = 89
F = 13.6
log MICM..avium 21d = – 0.321 ( 0.084) σ
– 0.262 ( 0.056) π + In
+ 1.658 ( 0.073)
s = 0.312 n = 110
(4)
+ 1.801 ( 0.078)
r = 0.765
s = 0.295
n = 94
F = 10.5
r = 0.739
F = 20.6

70 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
log MICM..kans. 21d = –0.680 ( 0.256) (σ1)CO
(parameter π) and electron-accepting properties of the
substituents (parameter σ).In conclusion, even though
salicylanilides are relatively simple compounds, they
have a broad spectrum of antimycobacterial effect, and
their structure is different from those of standard anti-
tuberculotics. Moreover, salicylanilides are superior to
INH as regards their activity against conditionally patho-
genic strains of mycobacteria. The mechanism of their
biological activity is currently being studied at the Hans
Knöll Institute at the University of Jena.
– 0.047 ( 0.275) (σ1)OH
–
– 0.539 ( 0.136) π1
–
– 0.202 ( 0.105) σ2
– 0.321 ( 0.061) π2
+ 0.029 ( 0.022)RM
+ 1.793 ( 0.084)
(5)
r = 0.752
s = 0.318
n = 89
F = 17.8
Acknowledgements
log MICM..avium 21d = 0.332 ( 0.222) (σ1)CO
This work was supported by the Ministry of Education of
the Czech Republic (project No. MSM 111600001) and
by the Grant Agency of the Czech Republic (grant No.
203/02/0082).We thank Mrs. D. Karlícšková (Department
of Pharmaceutical Chemistry and Drug Analysis, Facul-
ty of Pharmacy, Charles University) for performing ele-
mental analyses, Mrs. J. ZŠišzková (Department of Inor-
ganic and Organic Chemistry, Faculty of Pharmacy,
Charles University) for recording the IR spectra, and Ms
B.Janíšcková and M.Malíková (National Reference Labo-
ratory for Mycobacterium Kansasii, Regional Institute of
Hygiene, Ostrava) for their assistance in the antimyco-
bacterial assays.
+ 0.097 ( 0.238) (σ1)OH
–
– 0.820 ( 0.115) π1
–
– 0.331 ( 0.090) σ2
– 0.254 ( 0.060) π2
+ 0.074 ( 0.018)RM
+ 1.693 ( 0.078)
(6)
r = 0.700
s = 0.318
n = 94
F = 13.9
The results show that the influence of the substituents on
the phenyl ring in the amide part of the molecule is ap-
parent: antitubercular activity rises with increasing val-
ues of the Hammett constants σ and the hydrophobic
substituent constants π. The influence of the substitu-
ents from the acyl moiety is more complex.The correla-
tions using the indicator parameters (1–3) are more sig-
nificant than those including the substituent constants
(4–6). In determining the correlation equations for the
parameters of the acyl part of the molecule, we found
that more significant results can be obtained upon using
Experimental
Melting points were determined on a Kofler block and are un-
corrected. The IR spectra were measured in KBr pellets or in
CHCl3 solutions on a Nicolet Impact 400 apparatus; the wave-
numbers are given in cm–1.The NMR spectra were recorded on
a Varian Mercury-Vx BB 300 spectrometer operating at
300 MHz for 1H and 75 MHz for 13C in d6-DMSO. Chemical
shifts were recorded as δ values in ppm, and were indirectly ref-
erenced to tetramethylsilane via the solvent signal (7.26 for 1H
and 77.0 for 13C).The coupling constants J are given in Hz.Ele-
mental analyses were done on a CHNS-O CE elemental ana-
lyser (FISONS EA1110, Milano). Analyses of the C, H, N, S
content were within 0.4 % of the theoretical values. TLC was
performed on silica gel plates precoated with a fluorescent indi-
cator, Silufol UV 254 + 366 (Kavalier, Votice, the Czech Repub-
lic), in cyclohexane/acetone 3:1, to check the purity of the prod-
ucts.
–
the hydrophobic substituent constants π (with regard to
the hydroxy group) than the substituent constants π.The
differences in the regression coefficients of the σm and σp
parameters in correlations against various strains of my-
cobacteria are surprising. From the statistical point of
view, all equations can be considered proven on the level
of significance of 0.5. It has been shown [11] that upon
using the twofold serial dilution scale in antimicrobial
tests, the standard residual deviation up to the value of
0.3 can be explained by the methodical error in measure-
ment. For the acyl part of the molecule, the conclusion
can be drawn that the antimycobacterial activity increas-
General procedure for the preparation of salicylanilides
A suspension of a substituted salicylic acid (0.02 mol) and a
substituted aniline (0.02 mol) in chlorobenzene (100 mL) was
heated under reflux in the presence of PCl3 (0.01 mol) for three
hours.The reaction mixture was filtered while hot, and the sol-
vents evaporated. The product was crystallized from ethanol-
water (yields in the range 75–90 %, Table 1).
–
es with increasing lipophilicity (parameter π ) and de-
Microbiological assay
creasing molecular refraction (parameter MR).The influ-
ence of the electronic parameters in the acyl part of the
molecule cannot be evaluated in a simple way (parame-
ters σm and σp).As regards the amide moiety, the activity
is always favourably influenced by higher lipophilicity
For the evaluation of in vitro antimycobacterial activity of the
substances, the following strains were used: M. tuberculosis
CNCTC My 331/88, M. kansasii CNCTC My 235/80, M. avium
CNCTC My 330/88, obtained from the Czech National Collec-
tion of Type Cultures (CNCTC), National Institute of Public

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 53–71
Antimycobacterial Activity of Salicylanilides 71
[7] K. Waisser, J. Hladùvková, J. Kunes, L. Kubicová, V.
Klimesová, P. Karajannis, J. Kaustová, Chem. Pap. 2001,
56, 121–129.
Health, Prague. Antimycobacterial activity of the compounds
against these strains was determined in the Šula semisynthetic
medium (SEVAC, Prague).
[8] C. Hansch, A. Leo, Substituent Constants For Correlation
Analysis in Chemistry and Biology, John Wiley & Sons,
New York 1979.
The compounds were added to the medium in dimethyl sulfox-
ide solutions. The following concentrations were used: 1000,
500, 250, 125, 62, 31, 16, 8, and 4 µmol/L.The minimum inhibi-
tory concentrations were determined after incubation at 37 °C
for 14 days and 21 days (see Table 3). MIC was the lowest con-
centration of an antimycobacterially active substance (see the
above concentrations), at which the inhibition of the growth of
mycobacteria occurred.
°
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