Antimycobacterial and antifungal isosters of salicylamides

Article abstract of DOI:10.1002/ardp.200300725

A set of 40 derivatives of 3-hydroxypicolinic acid and 2-sulfanylbenzoic acid, isosteric to salicylanilides was synthesized. The compounds were evaluated for in vitro activity against Mycobacterium tuberculosis, Mycobacterium kansasii and Mycobacterium avium, Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, Trichosporon beigelii, Aspergillus fumigatus, Absidia corymbifera, Trichophyton mentagrophytes and Microsporum gypseum. Structure-activity relationships of antimycobacterial activity and antifungal activity against T. mentagrophytes and M. gypseum were analyzed by the Free-Wilson method. An increase in antimycobacterial activity was observed only for the sulfanylbenzoic acid derivatives, especially those with the benzyl moiety. The antifungal activity was not significant.

Full text of DOI:10.1002/ardp.200300725

322 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Karel Waissera,
Milan Pešinaa,
Pavel Holy´ a,
Antimycobacterial and Antifungal Isosters of
Salicylamides
Milan Poura,
A set of 40 derivatives of 3-hydroxypicolinic acid and 2-sulfanylbenzoic acid, isoster-
ic to salicylanilides was synthesized.The compounds were evaluated for in vitro ac-
tivity against Mycobacterium tuberculosis, Mycobacterium kansasii and Mycobac-
terium avium, Candida albicans, Candida tropicalis, Candida krusei, Candida gla-
brata, Trichosporon beigelii, Aspergillus fumigatus, Absidia corymbifera, Trichophy-
ton mentagrophytes and Microsporum gypseum. Structure-activity relationships of
antimycobacterial activity and antifungal activity against T. mentagrophytes and M.
gypseum were analyzed by the Free-Wilson method.An increase in antimycobacte-
rial activity was observed only for the sulfanylbenzoic acid derivatives, especially
those with the benzyl moiety. The antifungal activity was not significant.
Otakar Bureša,
Jiší Kuneša,
Všra Klimešováa,
Vladimír Buchtab,
Petra Kubanováb,
Jarmila Kaustovác
a
Department of Inorganic
and Organic Chemistry,
Faculty of Pharmacy,
Charles University,
Hradec Králové,
Keywords:Salicylanilides;Antimycobacterialactivity;Fungicidalactivity;Structure-
activity relationships; Tuberculostatics; Atypical mycobacterial strains
Czech Republic
Department of Biological
Received: September 4, 2002; Accepted: October 22, 2002 [FP725]
DOI 10.1002/ardp.200300725
b
and Medical Sciences,
Faculty of Pharmacy,
Charles University,
Hradec Králové, Czech
Republic
c
National Reference
Laboratory for
Mycobacterium kansasii,
Institute of Hygiene,
Ostrava, Czech Republic
picolinic and 2-sulfanylbenzoic acid, and some other iso-
steric structures as well (the derivatives derived from N-
benzylamines).Based on our preliminary studies, we as-
sumed that the compounds isosteric to salicylanilides
would also display antifungal activity [5, 6].
Introduction
The search for new antimycobacterially active com-
pounds is undoubtedly one of the important directions of
current pharmaceutical chemistry. Based on collabora-
tion with H. D. Stachel, we discovered a few new groups
of potential antituberculotics [1, 2]. During the research
on the antimycobacterial activity of salicylanilides [3], we
identified these compounds as broad spectrum antimy-
cobacterial substances, active not only against Myco-
bacterium tuberculosis, but also against other, condi-
tionally pathogenic strains.Salicylanilides are unlikely to
become effective chemotherapeutic agents themselves
due to their effect on mitochondrial oxidation. However,
they can serve as structural templates for the design of
potential drugs, as Macielag with coworkers have re-
cently found that they can act as inhibitors of two-compo-
nent regulatory systems in bacteria [4].The goal of this
work was to study selected isosteric modifications of
salicylanilides, in particular the anilides of 3-hydroxy-
Chemistry
All compounds are summarized in Scheme 1.Salicylani-
lides 1 were described in our previous paper [3]. Deriva-
tives 2–5 were prepared by the treatment of 3-hydroxy-
picolinic acid or 2-sulfanylbenzoic acid with substituted
anilines or benzylamines. The characteristics of the
compounds are presented in Table 1.
Microbiology
Antimycobacterial activity of compounds 2–5 was tested
in vitro against Mycobacterium tuberculosis, Mycobac-
terium kansasii, and Mycobacterium avium. In several
cases, the MIC values could not be determined due to
the limited solubility of the compounds. The antimyco-
bacterial activity of compounds 1 has been published re-
cently [3].
Correspondence: Karel Waisser, Faculty of Pharmacy,
Charles University, Heyrovského 1203, CZ 500 05 Hradec
Králové, Czech Republic. Phone: +42 049 5067246, Fax:
+42 049 5514330, e-mail: waisser@faf.cuni.cz.
© 2003 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
0365-6233/06–07/0322 $ 17.50+.50/0

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Antimycobacterial and antifungal isosters of salicylamides 323
Scheme 1
Table 1. Characteristics of the compounds.
Compound
R
Formula
ν C=O
M.p. (°C)
Mr
(cm–1)
Found
85–86.5
90.5–91.5
135–136
131–132
189–190
156
Ref.
2a
2b
2c
2d
2e
2f
H
C12H10N2O2
214.22
C13H12N2O2
228.25
C12H9ClN2O2
248.66
C12H9ClN2O2
248.66
C12H8Cl2N2O2
283.11
C12H9BrN2O2
293.11
C12H9FN2O2
232.21
C13H9F3N2O2
282.22
C12H9N3O4
259.22
C12H9N3O4
259.22
C13H12N2O3
244.24
C13H12N2O2
228.25
C14H14N2O2
242.27
C13H11ClN2O2
262.69
1658
1655
1656
1670
1664
1658
1656
1650
1662
1668
1655
1647
1652
1650
88–89 [8, 9]
89 [8, 9]
–
4-CH3
4-Cl
3-Cl
131 [8, 9]
3,4-Cl2
4-Br
180 [8]
189 [9]
150 [8, 9]
2g
2i
4-F
135–137
64–65
–
–
4-CF3
4-NO2
3-NO2
4-OCH3
H
2j
269–271
216–218
108–110
88–89
252–253 [8, 9]
–
2k
2l
108 [8, 9]
84 [8, 9]
80–81 [8, 9]
99–100 [8, 9]
3°
3b
3c
4-CH3
4–Cl
85–86
95–96

324 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Table 1. (continued).
Compound
R
Formula
ν C=O
M.p. (°C)
Mr
(cm–1)
1651
Found
Ref.
–
3d
3e
3f
3-Cl
C13H11ClN2O2
262.69
C13H10Cl2N2O2
297.14
C13H11BrN2O2
307.14
C13H11FN2O2
246.24
C14H11F3N2O2
296.24
C13H11N3O4
273.24
C13H11N3O4
273.24
C14H14N2O3
258.27
C13H11NOS
229.29
C14H13NOS
243.32
C13H10ClNOS
263.74
C13H10ClNOS
263.74
C13H9Cl2NOS
298.18
C13H10BrNOS
308.19
C13H10FNOS
247.28
C13H10FNOS
247.28
C14H13NO2S
259.32
C14H13NOS
243.32
C15H15NOS
257.35
C14H12ClNOS
277.76
C14H12ClNOS
277.76
C14H11Cl2NOS
312.21
C14H12BrNOS
322.21
C14H12FNOS
261.31
60.5–61
3,4-Cl2
4–Br
4-F
1648
1648
1649
1652
1649
1648
1646
1641
1645
1646
1649
1649
1650
1645
1648
1655
1633
1629
1630
1631
1631
1630
1628
1636
98
98 [8, 9]
103–104
69–71
–
–
–
3g
3i
4-CF3
4-NO2
3-NO2
4-OCH3
H
76–78
3j
116–117
106–108
86–87
128–129 [8, 9]
3k
3l
117–123 [8, 9]
94 [8, 9]
4°
102–104
124–126
113–116
118–120
161–163
102–104
129,5–131
87–90
109–110 [10]
4b
4c
4d
4e
4f
4-CH3
4-Cl
130–132 [11]
120–122 [11]
3-Cl
–
3,4-Cl2
4-Br
–
–
4g
4h
4l
4-F
–
3-F
–
4-OCH3
H
149–151
209–210
215–217
243–245
184–186
222–224
231–233
224–226
216–217
–
5a
5b
5c
5d
5e
5f
108 [12]
4-CH3
4-Cl
–
–
–
–
–
–
–
3-Cl
3,4-Cl2
4-Br
5g
5k
4-F
3-NO2
C14H12N2O3S
288.32

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Antimycobacterial and antifungal isosters of salicylamides 325
Table 2. 1H and 13C NMR spectra of new 3-hydroxypicolinanilides (2), 3-hydroxypicolinbenzylamides (3), 2-sulfanyl-
benzanilides (4) and 2-sulfanylbenzylamides (5).
Compounds
NMR, δ
1
1
1
2a
1H NMR (300 MHz, CDCl3) δ 11.96 (s, H, OH), 9.93 (bs, H, NH), 8.13 (dd, H J = 4.00 Hz, J =
1.78 Hz, H6), 7.75–7.69 (m, 2H, H2Ј, H6Ј), 7.45–7.33 (m, 4H, H5, H3Ј, H4Ј, H5Ј), 7.23–7.15 (m, 1H,
H4)
13C NMR (75 MHz, CDCl3) δ = 166.8, 158.2, 139.5, 136.7, 131.3, 129.2, 129.0, 126.5, 124.9, 120.1
2b
1H NMR (300 MHz, CDCl3) δ 12.02 (bs, 1H, OH), 9.86 (bs, 1H, NH), 8.11 (dd, 1H, J = 4.12 Hz, J =
1.65 Hz, H6), 7.64–7.56 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.41–7.32 (m, 2H, H4, H5), 7.24–7.16 (m AAЈ,
BBЈ, 2H, H3Ј, H5Ј), 2.35 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3) δ = 166.6, 158.2, 139.4, 134.6, 134.1, 131.4, 129.6, 128.8, 126.4, 120.0,
20.9
2c
2d
1H NMR (300 MHz, CDCl3) δ 11.78 (s, 1H, OH), 9.93 (bs, 1H, NH), 8.11 (dd, 1H, J = 4.23 Hz, J =
1.69 Hz, H6), 7.70–7.63 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.41–7.38 (m, 2H, H4, H5), 7.37–7.32 (m AAЈ,
BBЈ, 2H, H3Ј, H5Ј)
13C NMR (75 MHz, CDCl3) δ = 166.8, 158.2, 139.5, 135.3, 131.0, 129.9, 129.2, 129.1, 126.6, 121.2
1H NMR (300 MHz, CDCl3) δ 11.74 (s, 1H, OH), 9.95 (bs, 1H, NH), 8.12 (dd, 1H, J = 4.16 Hz, J =
1.25 Hz, H6), 7.85 (t, 1H, J = 2.08 Hz, H2Ј), 7.54 (ddd, 1H, J = 8.12 Hz, J = 2.08 Hz, J = 0.84 Hz, H6Ј),
7.44–7.25 (m, 3H, H4, H5, H5Ј), 7.15 (ddd, 1H, J = 8.12 Hz, J = 2.08 Hz, J = 0.84 Hz, H4Ј)
13C NMR (75 MHz, CDCl3) δ = 166.9, 158.3, 139.6, 137.9, 134.8, 130.9, 130.1, 129.2, 126.6, 124.9,
120.1, 117.9
2e
1H NMR (300 MHz, CDCl3) δ 11.62 (s, 1H, OH), 9.96 (bs, 1H, NH), 8.12 (dd, 1H J = 4.16 Hz, J =
1.67 Hz, H6), 7.97 (d, 1H, J = 2.08 Hz, H2Ј), 7.54 (dd, 1H, J = 8.74 Hz, J = 2.50 Hz, H6Ј), 7.45 (d over-
lapped, 1H, J = 8.74 Hz, H5Ј), 7.42 (dd overlapped, 1H, J = 8.84 Hz, J = 4.16 Hz, H5), 7.37 (dd, 1H J =
8.84 Hz, J = 1.67 Hz, H4)
13C NMR (75 MHz, CDCl3) δ = 166.9, 158.3, 139.7, 136.2, 133.0, 130.7, 130.7, 129.4, 128.1, 126.7,
121.6, 119.1
2f
1H NMR (300 MHz, CDCl3) δ 11.77 (s, 1H, OH), 9.93 (bs, 1H, NH), 8.12 (dd, 1H, J = 4.17 Hz, J =
1.65 Hz, H6), 7.65–7.59 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 7.53–7.47 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.41 (dd,
1
1H, J = 8.45 Hz, J = 4.17 Hz, H5), 7.35 (dd, H, J = 8.45 Hz, J = 1.65 Hz, H4)
13C NMR (75 MHz, CDCl3) δ = 166.8, 158.2, 139.6, 135.8, 132.1, 131.0, 129.2, 126.6, 121.5, 117.6
2g
1H NMR (300 MHz, DMSO) δ 11.88 (bs, 1H, OH), 9.93 (s, 1H, NH), 8.12 (dd, 1H, J = 4.12 Hz, J =
1.65 Hz, H6), 7.73–7.63 (m, 2H, H2Ј, H6Ј), 7.45–7.33 (m, H4, H5), 7.14–7.05 (m AAЈ, BBЈ, 2H, H3Ј,
H5Ј)
13C NMR (75 MHz, DMSO) δ 166.6, 161.3 and 158.1 (J = 244.5 Hz), 158.3, 139.4, 132.8 and 132.7
(J = 2.9 Hz), 131.0, 129.1, 126.7, 121.9 and 121.8 (J = 8.0 Hz), 116.0 and 115.7 (J = 22.6 Hz)
2i
1H NMR (300 MHz, DMSO) δ 11.84 (bs, 1H, OH), 11.20 (s, 1H, NH), 8.27 (d, 1H, J = 4.12 Hz, H6),
8.12–8.05 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 7.77–7.71 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.61 (dd, J = 8.51 Hz,
1
J = 4.12 Hz, H5), 7.5 (dd, H, J = 8.52 Hz, J = 1.38 Hz, H4)
13C NMR (75 MHz, DMSO) δ 168.0, 157.8, 141.2, 140.3, 130.0, 126.6, 126.3, 126.1 (q, J = 3.7 Hz),
125.3 (q, J = 272.6 Hz), 124.8 (q, J = 32.1 Hz), 121.5
2j
1H NMR (300 MHz, DMSO) δ 11.64 (bs, 1H, OH), 11.35 (bs, 1H, NH), 8.31–8.22 (m AAЈBBЈover-
lapped, 3H, H6, H3Ј, H5Ј), 8.18–8.11 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.65–7.58 (m, 1H, H5), 7.53–7.47
(m, 1H, H4)
13C NMR (75 MHz, DMSO) δ 168.1, 157.7, 143.8, 143.4, 140.4, 131.5, 130.1, 126.7, 124.8, 121.3
2k
1H NMR (300 MHz, DMSO) δ 11.74 (bs, 1H, NH), 11.33 (bs, 1H, OH), 8.88 (t, 1H, J = 2.20 Hz, H2Ј),
8.29–8.21 (m, 2H, H6, H4Ј), 8.02–7.97 (m, 1H, H6Ј), 7.69–7.57 (m, 2H, H5, H5Ј), 7.49 (dd, 1H, J =
8.52 Hz, J = 1.37 Hz, H4)
13C NMR (75 MHz, DMSO) δ 168.0, 157.8, 148.0, 140.3, 138.8, 131.3, 130.2, 130.0, 127.6, 126.6,
119.3, 115.7

326 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Table 2. (continued).
Compounds
NMR, δ
2l
3a
3b
3c
3d
3e
3f
1H NMR (300 MHz, CDCl3) δ 12.09 (bs, 1H, OH), 9.89 (s, 1H, NH), 8.12 (dd, 1H, J = 3.85 Hz, J =
1.65 Hz, H6), 7.67–7.59 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.44–7.34 (m, H4, H5), 7.03–6.85 (m AAЈ, BBЈ,
2H, H3Ј, H5Ј), 3.82 (s, 3H, OCH3)
13C NMR (75 MHz, CDCl3) δ 166.27, 158.3, 156.9, 139.2, 131.2, 129.8, 128.9, 126.8, 121.8, 114.3,
55.5
1H NMR (300 MHz, DMSO) δ 12.51 (bs, 1H, OH), 9.76 (t, 1H, J = 6.18 Hz, NH), 8.16 (dd, 1H, J =
4.39 Hz, J = 1.37 Hz, H6), 7.53 (dd, 1H, J = 8.52 Hz, J = 4.4 Hz, H5), 7.41 (dd, 1H, J = 8.52 Hz, J =
1.38 Hz, H4), 7.37–7.19 (m, 5H, H2Ј, H3Ј, H4Ј, H5Ј, H6Ј), 4.49 (d, 2H, J = 6.59 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 168.9, 157.5, 140.1, 139.0, 131.5, 129.5, 128.6, 127.7, 127.2, 126.1,
42.3
1H NMR (300 MHz, CDCl3) δ 12.2 (bs, 1H, OH), 8.38 (bs, 1H, NH), 8.03 (dd, 1H, J = 3.57 Hz, J =
2.2 Hz, H6), 7.35–7.32 (m, 2H, H4, H5), 7.29–7.23 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.20–7.14 (m AAЈ,
BB, 2H, H3Ј, H5Ј), 4.6 (d, 2H, J = 6.32 Hz, CH2), 2.35 (s, 3H, CH3)
13C NMR (75 MHz, CDCl3) δ 168.4, 157.8, 139.28, 137.4, 134.3, 131.3, 129.4, 128.6, 127.8, 126.3,
42.8, 21.1
1H NMR (300 MHz, DMSO) δ 12.45 (bs, 1H, OH), 9.80 (bs, 1H, NH), 8.16 (dd, 1H, J = 3.44 Hz, J =
1.37 Hz, H6), 7.52 (dd, 1H, J = 7.56 Hz, J = 3.44 Hz, H5), 7.41 (dd, 1H, J = 7.56 Hz, J = 1.38 Hz,
H4), 7.38–7.32 (m, 4H, H2Ј, H3Ј, H5Ј, H6Ј), 4.47 (d, 2H, J = 6.31 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 169, 157.5, 140.1, 138.1, 131.7, 131.4, 129.6, 129.5, 128.5, 126.2,
41.65
1H NMR (300 MHz, DMSO) δ 12.41 (bs, 1H, OH), 9.81 (t, 1H, J = 6.32 Hz, NH), 8.16 (dd, 1H, J =
4.39 Hz, J = 1.38 Hz, H6), 7.52 (dd, 1H, J = 8.52 Hz, J = 4.4, H5), 7.41 (dd, 1H, J = 8.51 Hz, J =
1.38 Hz, H4), 7.38–7.27 (m, 5H, H2Ј, H4Ј, H5Ј, H6Ј), 4.49 (d, 2H, J = 6.59 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 169.1, 157.5, 141.6, 140.1, 133.2, 131.3, 130.5, 129.5, 127.5, 127.2,
126.3, 126.15, 41.8
1H NMR (300 MHz, DMSO) δ 12.45 (bs, 1H, OH), 9.83 (t, 1H, J = 6.04 Hz, NH), 8.16 (dd, 1H, J =
4.39 Hz, J = 1.37 Hz, H6), 7.62–7.49 (m, 3H, H2Ј, H5Ј, H6Ј), 7.41 (dd, 1H, J = 8.38 Hz, J = 1.37 Hz,
H5), 7.32 (dd, J = 8.38 Hz, J = 1.37 Hz, H4), 4.47 (d, 2H, J = 6.32 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 169.1, 157.5, 140.3, 140.2, 131.3, 131.1, 130.8, 129.8, 129.7, 129.5,
128.1, 126.2, 41.3
1H NMR (300 MHz, DMSO) δ 12.44 (bs, 1H, OH), 9.80 (t, 1H, J = 6.32 Hz, NH), 8.16 (dd, 1H, J =
4.12 Hz, J = 1.37 Hz, H6), 7.55–7.52 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.51 (dd, 1H, J = 6.05 Hz, J =
1.37 Hz, H5), 7.41 (dd, 1H, J = 8.51 Hz, J = 1.37 Hz, H4), 7.32 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 4.45 (d,
2H, J = 6.31 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 169, 157.5, 140.1, 138.5, 131.4, 131.4, 129.9, 129.4, 126.2, 120.3, 41.7
1H NMR (300 MHz, DMSO) δ 12.48 (bs, 1H, OH), 9.78 (t, 1H, J = 6.32 Hz, NH), 8.15 (dd, 1H, J =
4.39 Hz, J = 1.38 Hz, H6), 7.43–7.34 (m, 4H, H4, H5, H2Ј, H6Ј), 7.19–7.09 (m, 2H, H3Ј, H5Ј), 4.46
(d, 2H, J = 6.31 Hz, CH2)
3g
3i
13C NMR (75 MHz, DMSO) δ 168.9, 163.1 and 159.9 (J = 242.5 Hz), 157.5, 140.1, 135.2 and 135.2
(J = 2.9 Hz), 131.4, 129.8 and 129.7 (J = 8.1 Hz), 129.4, 126.1, 115.4 and 115.1 (J = 21.2 Hz)
1H NMR (300 MHz, DMSO) δ 12.39 (bs, 1H, OH), 9.88 (t, 1H, J = 6.32 Hz, NH), 8.17 (dd, 1H, J =
4.12 Hz, J = 1.37 Hz, H6), 7.73–7.65 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 7.6–7.49 (m AAЈ, BBЈ overlapped,
3H, H5, H2Ј, H6Ј), 7.41 (dd, 1H, J = 8.51 Hz, J = 1.37 Hz, H4), 4.57 (d, 2H, J = 6.32 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 169.2, 157.5, 143.9, 140.2, 131.4, 129.5, 128.3, 127.8 (q, J = 31.8 Hz),
126.2, 125.5 (q, 3.8 Hz), 124.5 (q, J = 272.0 Hz), 42.0
3j
1H NMR (300 MHz, DMSO) δ 12.34 (bs, 1H, OH), 9.93 (t, 1H, J = 6.32 Hz, NH), 8.24–8.13 (m, 3H,
H6, H3Ј, H5Ј), 7.63–7.5 (m, 3H, H5, H2Ј, H6Ј), 7.41 (dd, 1H, J = 8.51 Hz, J = 1.37 Hz, H4), 4.61 (d,
2H, J = 6.31 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 169.26, 157.5, 147, 146.8, 140.2, 131.3, 129.6, 128.6, 126.2, 123.8,
41.9

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Antimycobacterial and antifungal isosters of salicylamides 327
Table 2. (continued).
Compounds
NMR, δ
3k
1H NMR (300 MHz, DMSO) δ 12.33 (bs, 1H, OH), 9.95 (t, 1H, J = 6.32 Hz, NH), 8.23–8.2 (m, 1H,
1H2Ј), 8.18 (dd, 1H, J = 4.12 Hz, J = 1.38 Hz, H6), 8.15–8.08 (m, 1H, H4Ј), 7.8 (d, 1H, J = 7.69 Hz,
1H, H6Ј), 7.67–7.58 (m, 1H, H5Ј), 7.54 (dd, J = 8.51 Hz, J = 4.12 Hz, H5), 7.41 (dd, 1H, J = 8.51 Hz,
J = 1.37 Hz, H4), 4.61 (d, 2H, J = 6.32 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 169.2, 157.5, 148, 141.4, 140.2, 134.5, 131.3, 130.2, 129.6, 126.2,
122.4, 122.3, 41.7
3l
1H NMR (300 MHz, DMSO) δ 12.56 (bs, 1H, OH), 9.67 (t, 1H, J = 6.32 Hz, NH), 8.14 (dd, 1H, J =
4.40 Hz, J = 1.37 Hz, H6), 7.51 (dd, 1H, J = 8.52 Hz, J = 4.40 Hz, H5), 7.40 (dd, J = 8.52 Hz, J =
1.33 Hz, H4), 7.31–7.24 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 6.91–6.84 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 4.41 (d,
2H, J = 6.60 Hz, CH2), 3.70 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO) δ 168.8, 158.6, 157.5, 140, 131.5, 131, 129.4, 129.2, 126.1, 113.9, 55.3,
41.7
4a
4b
1H NMR (300 MHz, DMSO) δ 10.37 (bs, 1H, NH), 7.77–7.67 (m, 2H, H2Ј, H6Ј), 7.60 (dd, 1H, J =
7.42 Hz, J = 1.37 Hz, H6), 7.49 (dd, 1H, J = 7.42 Hz, J = 1.37 Hz, H3), 7.39–7.30 (m, 3H, H3Ј, H4Ј,
H5Ј), 7.24 (dt, 1H, J = 7.42, J = 1.37 Hz, H4), 7.14–7.06 (m, 1H, H5), 5.24 (bs, 1H, SH)
13C NMR (75 MHz, DMSO) δ 166.7, 139.3, 134.3, 133.3, 130.7, 128.9, 128.8, 124.9, 124.0, 120.2
1H NMR (300 MHz, DMSO) δ 10.30 (bs, 1H, NH), 7.65–7.56 (m, 3H, H6, H2Ј, H6Ј), 7.51–7.45 (m,
1H, H3), 7.34 (dt, 1H, J = 7.69 Hz, J = 1.37 Hz, H4), 7.23 (dt, 1H, J = 7.70 Hz, J = 1.38 Hz, H5),
7.18–7.11 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 5.33 (bs, 1H, SH), 2.27 (s, 3H, CH3)
13C NMR (75 MHz, DMSO) δ 166.5, 136.8, 134.3, 133.4, 132.9, 130.7, 129.3, 128.7, 124.9, 120.3,
120.2, 20.7
4c
4d
1H NMR (300 MHz, DMSO) δ 10.52 (bs, 1H, NH), 7.81–7.72 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.61 (dd,
1H, J = 7.69 Hz, J = 1.37 Hz, H6), 7.50 (dd, 1H, J = 7.69 Hz, J = 1.37 Hz, H3), 7.43–7.37 (m AAЈ,
BBЈ, 2H, H3Ј, H5Ј), 7.35 (dt overlapped, 1H, J = 7.69 Hz, J = 1.37 Hz, H4), 7.24 (dt, 1H, J = 7.69 Hz,
J = 1.37 Hz, H5), 5.26 (bs, 1H, SH)
13C NMR (75 MHz, DMSO) δ 166.7, 138.2, 133.9, 133.5, 130.9, 130.8, 128.9, 128.8, 127.5, 124.9,
121.7
1H NMR (300 MHz, DMSO) δ 10.55 (bs, 1H, NH), 7.93 (t, 1H, J = 1.92 Hz, H2Ј), 7.67–7.58 (m, 2H,
H6, H6Ј), 7.51 (dd, 1H, J = 7.97 Hz, J = 1.10 Hz, H3), 7.41–7.32 (m, 2H, H4, H5Ј), 7.25 (dt, 1H, J =
7.41 Hz, J = 1.10 Hz, H5), 7.16 (ddd, 1H, J = 7.97 Hz, J = 2.20 Hz, J = 1.10 Hz, H4Ј), 5.26 (bs, 1H,
SH)
13C NMR (75 MHz, DMSO) δ 166.9, 140.7, 133.8, 133.5, 133.2, 131.0, 130.8, 130.6, 128.8, 125.0,
123.7, 119.5, 118.5
4e
4f
1H NMR (300 MHz, DMSO) δ 10.66 (bs, 1H, NH), 8.11 (s, 1H, J = 2.19 Hz, H2Ј), 7.71–7.57 (m, 3H,
H6, H5Ј, H6Ј), 7.51 (dd, 1H, J = 7.42 Hz, J = 0.83 Hz, H3), 7.37 (dt, 1H, J = 7.42 Hz, J = 1.38 Hz,
H4), 7.25 (dt, 1H, J = 7.41 Hz, J = 1.38 Hz, H5), 5.27 (bs, 1H, SH)
13C NMR (75 MHz, DMSO) δ 166.9, 139.4, 133.8, 133.5, 131.2, 131.1, 130.9, 130.9, 128.9, 125.4,
125.0, 121.2, 120.1
1H NMR (300 MHz, DMSO) δ 10.50 (bs, 1H, NH), 7.75–7.67 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.61 (dd,
1H, J = 7.42 Hz, J = 1.37 Hz, H6), 7.56–7.47 (m AAЈ, BBЈ, 2H, H3Ј, H5Ј), 7.36 (dt, 1H, J = 7.42 Hz,
J = 1.37 Hz, H4), 7.24 (dt, 1H, J = 7.41 Hz, J = 1.37 Hz, H5), 5.26 (bs, 1H, SH)
13C NMR (75 MHz, DMSO) δ 166.7, 138.6, 133.9, 133.5, 131.7, 130.9, 130.8, 128.8, 125.0, 122.0,
115.6
4g
1H NMR (300 MHz, DMSO) δ 10.44 (bs, 1H, NH), 7.81–7.70 (m, 2H, H2Ј, H6Ј), 7.61 (dd, 1H, J =
7.56 Hz, J = 1.37 Hz, H6), 7.49 (dd, 1H, J = 7.56 Hz, J = 1.37 Hz, H3), 7.35 (dt, 1H, J = 7.56 Hz, J =
1.37 Hz, H4), 7.25 (dt, 1H, J = 7.56 Hz, J = 1.37 Hz, H5), 7.23–7.14 (m, 2H, H3Ј, H5Ј), 5.25 (bs, 1H,
SH)
13C NMR (75 MHz, DMSO) δ 166.5, 158.5 (d, J = 240.5 Hz), 135.6 (d, J = 2.6 Hz), 134.0, 133.4, 130.8,
128.8, 124.9, 122.0 (d, J = 7.7 Hz), 115.5 (d, J = 22.3 Hz)

328 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Table 2. (continued).
Compounds
NMR, δ
4h
1H NMR (300 MHz, DMSO) δ 10.59 (bs, 1H, NH), 7.80–7.65 (m, 1H, H2Ј), 7.61 (dd, 1H, J = 7.70 Hz,
J = 1.38 Hz, H6), 7.54–7.45 (m, 2H, H3, H6Ј), 7.44–7.32 (m, 2H, H4, H5Ј), 7.25 (dt, 1H, J = 7.70 Hz,
J = 1.38 Hz, H5), 6.98–6.89 (m, 1H, H4Ј), 5.26 (bs, 1H, SH)
13CNMR(75MHz, DMSO)δ 166.9, 166.1, 162.3(d, J= 241.1Hz), 141.0(d, J=10.9Hz), 133.7(d, J =
23.8 Hz), 130.9 (d, J = 7.5 Hz), 130.6 (d, J = 9.7 Hz), 128.8, 125.0, 115..9 (d, J = 2.6 Hz), 110.4 (d, J =
20.9 Hz), 107.0, 106.6
4l
1H NMR (300 MHz, DMSO) δ 10.25 (bs, 1H, NH), 7.68–7.60 (m AAЈ, BBЈoverlapped, 2H, H2Ј, H6Ј),
7.59 (dd overlapped, 1H, J = 7.56 Hz, J = 1.10 Hz, H6), 7.48 (dd, 1H, J = 7.56 Hz, J = 1.10 Hz, H3),
7.33 (dt, 1H, J = 7.56 Hz, J = 1.10 Hz, H4), 7.23 (dt, 1H, J = 7.56 Hz, J = 1.10 Hz, H5), 7.00–6.87 (m
AAЈ, BBЈ, 2H, H3Ј, H5Ј), 5.25 (bs, 1H, SH), 3.73 (s, 3H, OCH3)
13C NMR (75 MHz, DMSO) δ 166.2, 155.8, 134.2, 133.4, 132.4, 130.7, 130.6, 128.7, 124.9, 121.7,
114.0, 55.4
5a
5b
1H NMR (300 MHz, DMSO) δ 9.21 (t, 1H, J = 6.04 Hz, NH), 7.71 (dd, 1H, J = 7.56 Hz, J = 1.10 Hz,
H6), 7.65 (d, 1H, J = 7.56 Hz, H3), 7.48–7.31 (m, 7H, H4, H5, H2Ј, H3Ј, H4Ј, H5Ј, H6Ј), 4.50 (d, 1H, J
= 6.04 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 167.1, 139.5, 137.1, 133.8, 131.4, 128.6, 128.2, 127.5, 127.1, 126.2,
126.0, 42.8
1H NMR (300 MHz, DMSO) δ 9.16 (t, 1H, J = 6.04 Hz, NH), 7.68 (dd, 1H, J = 7.76 Hz, J = 1.30 Hz,
H6), 7.64 (dd, 1H, J = 7.76 Hz, J = 1.30 Hz, H3), 7.42 (dt, 1H, J = 7.76 Hz, J = 1.30 Hz, H4), 7.30 (dt
overlap, 1H, J = 7.76 Hz, J = 1.30, H5) 7.28–7.22 (m AAЈBBЈoverlap, 2 H, H2Ј, H6Ј), 7.17–7.12 (m
AAЈ, BBЈ, 2H, H3Ј, H5Ј), 4.44 (d, 2H, J = 6.05 Hz, CH2), 2.27 (s, 3H, CH3)
13C NMR (75 MHz, DMSO) δ 167.1, 137.1, 136.5, 136.1, 133.9, 131.4, 129.1, 128.2, 127.5, 126.2,
125.9, 42.6, 20.9
5c
5d
1H NMR (300 MHz, DMSO) δ 9.24 (t, 1H, J = 5.77 Hz, NH), 7.71 (dd, 1H, J = 7.83 Hz, J = 0.96 Hz,
H6), 7.63 (dd, 1H, J = 7.83 Hz, J = 0.96 Hz, H3), 7.49–7.35 (m, 5H, H4, H2Ј, H3Ј, H5Ј, H6Ј), 7.35–
7.27 (m, 1H, H5), 4.47 (d, 2H, J = 6.05 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 167.2, 128.6, 137.1, 133.7, 131.7, 131.5, 129.4, 128.5, 128.2, 126.3,
126.0, 42.2,
1H NMR (300 MHz, DMSO) δ 9.26 (t, 1H, J = 6.05 Hz, NH), 7.71 (dd, 1H, J = 7.69 Hz, J = 1.10 Hz,
H6), 7.67–7.62 (m, 1H, H3), 7.49–7.26 (m, 6H, H4, H5, H2Ј, H4Ј, H5Ј, H6Ј), 4.48 (d, 2H, J = 6.05 Hz,
CH2)
13C NMR (75 MHz, DMSO) δ 167.3, 142.1, 137.0, 133.7, 133.2, 131.5, 130.5, 128.2, 127.3, 127.1,
126.3, 126.2, 126.0, 42.4
5e
5f
1H NMR (300 MHz, DMSO) δ 9.26 (t, 1H, J = 6.05 Hz, NH), 7.71 (dd, 1H, J = 7.69 Hz, J = 1.10 Hz,
H6), 7.66–7.58 (m, 3H, H3, H2Ј, H5Ј), 7.48–7.28 (m, 4H, H4, H5, H6Ј), 4.48 (d, 2H, J = 6.05 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 167.3, 140.8, 137.0, 133.6, 131.6, 131.1, 130.8, 129.6, 129.5, 128.3,
127.9, 126.4, 126.0, 41.9
1H NMR (300 MHz, DMSO) δ 9.25 (t, 1H, J = 6.04 Hz, NH), 7.70 (dd, 1H, J = 7.76 Hz, J = 1.10 Hz,
H6), 7.65–7.60 (m, 1H, H3), 7.57–7.50 (m AAЈ, BBЈ, 2H, H2Ј, H6Ј), 7.44 (dt, 1H, J = 7.76 Hz, J =
1.10, H4) 7.37–7.27 (m, 3H, H5, H3Ј, H5Ј), 4.45 (d, 2H, J = 6.05 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 167.2, 139.0, 137.1, 133.6, 131.5, 131.4, 129.8, 129.7, 128.2, 126.3,
120.1, 42.3
5g
1H NMR (300 MHz, DMSO) δ 9.22 (t, 1H, J = 6.04 Hz, NH), 7.70 (dd, 1H, J = 7.69 Hz, J = 1.10 Hz,
H6), 7.63 (dd, 1H, J = 7.69 Hz, J = 1.10 Hz, H3), 7.47–7.36 (m, 3H, H4, H2Ј, H6Ј), 7.30 (dt, 1H, J =
7.69 Hz, J = 1.10, H5) 7.22–7.11 (m, 2H, H3Ј, H5Ј), 4.47 (d, 2H, J = 6.04 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 167.1, 161.5 (d, J = 242.2 Hz), 137.1, 135.7 (d, J =2.9 Hz), 133.8, 131.5,
129.5 (d, J = 8.0 Hz), 128.2, 126.3, 126.0, 115.3 (d, J = 21.1 Hz), 42.2

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Antimycobacterial and antifungal isosters of salicylamides 329
Table 2. (continued).
Compounds
NMR, δ
5k
1H NMR (300 MHz, DMSO) δ 9.36 (t, 1H, J = 6.05 Hz, NH), 8.25 (t, 1H, J = 1.65 Hz, H2Ј), 8.16–8.10
(m, 1H, H6Ј), 7.87–7.81 (m, 1H, H4Ј), 7.73 (dd, 1H, J = 7.41 Hz, J = 1.10 Hz, H6), 7.69–7.61 (m, 2H,
H3, H5Ј), 7.45 (dt, 1H, J = 7.42 Hz, J = 1.10 Hz, H4), 7.33 (dt, 1H, J = 7.42 Hz, J = 1.10, H5), 4.62 (d,
2H, J = 6.05 Hz, CH2)
13C NMR (75 MHz, DMSO) δ 167.4, 148.0, 141.9, 137.0, 134.3, 133.5, 131.6, 130.2, 128.3, 126.3,
126.0, 122.2, 122.1, 42.3
Compounds 1–5 were evaluated for in vitro antifungal
activity against nine strains of human pathogenic fungi.
With the exceptions of Trichophyton mentagrophytes
445 and Microsporum gypseum 27339, the antifungal
activities of the derivatives were low and, in some cases,
the values of minimum inhibitory concentration (MIC)
could not be determined due to the limited solubility of
the compounds. For this reason, quantitative structure-
antifungal activity relationships were studied only for the
above dermatophytic strains.
Results and discussion
The characteristics of the compounds are presented in
Table 1. The compounds were characterized by NMR
and IR spectroscopy. In the IR spectra, the carbonyl vi-
bration ν(C=O) was in the region 1650–1670 cm–1 for pi-
colinanilides (2), in the region 1652–1646 cm–1 for 2-hy-
droxypicolinbenzylamides (3), in the region 1641–
1655 cm–1 for 2-sulfanylbenzanilides (4), and in the re-
gion 1628–1636 cm–1 for N-benzyl-2-sulfanylbenz-
amides (5). For NMR data, see Table 2.
The antimycobacterial activity is given in Table 3. The
most active compounds could be considered superior to
INH, given by their broad spectrum of activity. Thus, we
have found 4 groups of potentially new antituberculotics,
i.e. 3-hydroxypicolinanilides 2, N-benzyl-3-hydroxypico-
Calculation
Quantitative structure-activity relationships were deter-
mined by the Free-Wilson method modified according to
Fujita and Ban [7].
Table 3. Antimycobacterial activities (MIC(ìmol/L)) of substances 1–5.
Compounds
MIC (µmol/L)
Incubation time 14 d/21 d
R
M. tuberculosis
M. kansasii
M. avium
1ab
1bb
1cb
1db
1eb
1fb
1gb
1hb
1ic
H
4-CH3
4-Cl
3-Cl
3,4-Cl
4-Br
62.5/62.5
62.5/62.5
31/31
16/16
28/8
16/31
62.5/62.5
31/31
8/8
8/16
16/166
62.5/62.5
250/250
125/a
125/250
62.5/125
31/31
8/8
4/8
16/31
62.5/125
62.5/62.5
16/16
16/16
2.5/a
250/250
125/125
62.5/62.5
62.5/62.5
62.5/125
31/62.5
31/31
31/31
16/31
31/31
4-F
3-F
31/62.5
62.5/62.5
16/32
31/31
31/a
62.5/125
125/250
62.5/125
32/62.5
4-CF3
4-NO2
3-NO2
4-OCH3
H
1jb
1kb
1lb
2a
2b
4-CH3
4-Cl
2c
62.5/62.5

330 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Table 3. (continued).
Compounds
MIC (µmol/L)
Incubation time 14 d/21 d
R
M. tuberculosis
M. kansasii
M. avium
2d
2e
2f
2g
2i
2j
2k
2l
3-Cl
3,4-Cl2
4-Br
62.5/62.5
a/a
62.5/62.5
128/125
32/62.5
a/a
62.5/62.5
a/a
32/62.5
a/a
32/62.5
62.5/125
62.5/62.5
a/a
62.5/62.5
122/125
62.5/62.5
>1000/>1000
a/>1000
32/62.5
125/500
62.5/125
32/62.5
32/62.5
16/32
32/62.5
32/62.5
62.5/62.5
125/250
62.5/125
125/125
32/32
16/16
16/16
62.5/62.5
32/32
16/16
32/32
62.5/62.5
16/16
4-F
4-CF3
4-NO2
3-NO2
4-OCH3
H
4-CH3
4-Cl
3-Cl
3,4-Cl2
4-Br
4-F
4-CF3
4-NO2
3-NO2
4-OCH3
H
4-CH3
4-Cl
3-Cl
3,4-Cl2
4-Br
4-F
3-F
4-OCH3
H
4-CH3
4-Cl
3-Cl
a/a
a/a
a/a
125/125
500/1000
a/a
125/125
125/125
62.5/62.5
a/a
250/500
125/125
125/250
250/500
a/a
32/32
16/16
32/32
32/32
62.5/62.5
62.5/62.5
16/16
32/32
16/16
8/8
3°
250/500
125/125
62.5/125
62.5/125
32/62.5
32/62.5
125/2
62.5/125
125/125
125/250
a/a
62.5/62.5
32/32
32/32
62.5/62.5
62.5/62.5
16/32
62.5/62.5
62.5/62.5
16/16
3b
3c
3d
3e
3f
3g
3i
3j
3k
3l
4°
4b
4c
4d
4e
4f
4g
4h
4l
5a
5b
5c
5d
5e
5f
2/2
4/4
4/4
4/4
4/8
4/4
8/8
16/16
16/16
8/16
16/32
8/16
8/8
8/8
8/16
32/32
8/8
3,4-Cl2
4-Br
5g
5j
4-F
4-NO2
3-NO2
8/8
8/16
8/8
½
2/4
8/8
8/8
8/16
16/16
16/16
250/250
5k
INH
250/250
a: MIC not determined due to a low solubility of the compound
b: data were taken from Ref. [3]
linamides 3, 2-sulfanylbenzanilides 4, and N-benzyl-2-
sulfanylbenzamides 5. Quantitative structure-antimyco-
bacterial activity relationships were determined by the
Free-Wilson method (seeTable 4).As regards the substi-
tution on the phenyl ring in the amide part of the mole-
cule, the introduction of lipophilic substituents, e.g.
4-bromo-, 3-chloro-, 4-chloro-, and 4-trifluoromethyl
moiety led to the highest increase of antimycobacterial
activity. The activity increased upon isosteric replace-
ment of the hydroxy group in the acyl part with the sulfa-
nyl substituent.The derivatives of picolinic acid 2 and 3,
however, were less active than the corresponding deriv-

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Antimycobacterial and antifungal isosters of salicylamides 331
Table 4. Structure – antimycobacterial activity relationships.Results of the Free-Wilson analysis, modification accord-
ing to Fujita and Ban (incubation 14 and 21 d).
⌬ log MIC (µmol/L)
Fragment
M. tuberculosis
M. kansasii
M. avium
331/88
235/80
330/88
14d
21d
14d
21d
14d
0
21d
OH
SH
0
0
0
0
0
–0.1425
–0.1728
–0.265
–0.3574
–0.0593
–0.1913
C
N
0
0
0
0
0
0
0.7708
0.8024
0.557
0.4776
0.484
0.5093
NH
0
0
0
0
0
0
NHCH2
–0.0919
–0.0426
–0.4228
–0.444
–0.1889
–0.1313
HЈ
4Ј-CH3
4Ј-Cl
3Ј-Cl
3Ј,4Ј-Cl2
4Ј-Br
0
0
0
0
0
0
–0.2443
–0.301
–0.2557
–0.3612
–0.3584
–0.2778
–0.2919
–0.1806
–0.1004
–0.6069
–0.2643
–0.2643
–0.1969
1.6848
–0.1814
–0.2998
–0.299
–0.4887
–0.3572
–0.061
0.2548
–0.3035
–0.0858
0.0098
0.0228
1.6717
–0.0976
–0.2769
–0.2762
–0.3912
–0.2769
0.0228
0.2466
–0.1595
–0.0668
0.0073
0.1092
1.7261
–0.1786
–0.2969
–0.299
–0.2725
–0.4755
–0.1806
0.1207
–0.3639
–0.1211
–0.1211
–0.0246
1.7028
–0.2396
–0.36
–0.3585
–0.2205
–0.3187
–0.1806
–0.0756
–0.6264
–0.4921
–0.2914
–0.1566
1.6449
–0.3018
–0.223
–0.4202
–0.1212
–0.0143
–0.4003
–0.325
–0.1306
–0.0072
1.9039
4Ј-F
3Ј-F
4Ј–CF3
4Ј-NO2
3Ј-NO2
4Ј-OCH3
µ0
R = 0.8363 R = 0.8215
R = 0.8117 R = 0.8081
R = 0.7787 R = 0.8405
s = 0.334
F = 5.15
n = 46
s = 0.3684
F = 4.45
n = 45
s = 0.3728
F = 4.69
n = 49
s = 0.385
F = 4.57
n = 48
s = 0.265
F = 3.52
n = 47
s = 0.2529
F = 5.33
n = 46
Table 5. Structure – antifungal activity relationships. Results of the Free-Wilson analysis, modification according to
Fujita and Ban (incubation 72 and 120 h).
⌬ log MIC (µmol/L)
Fragment
M. gypseum
T. mentagrophytes
120 h
72 h
120 h
72 h
OH
SH
0
0
0
0
0.2822
–0.0645
–0.2527
0.0251
C
N
0
0
0
0
0.5644
0.473
0.7739
0.7525
HЈ
0
0
0
0
4Ј-CH3
4Ј-Cl
3Ј-Cl
8.9E-16
–0.3011
–0.4014
–0.1505
–0.6021
–0.4515
–0.1003
–0.8151
–0.5142
–2E-16
–0.5185
–0.4431

332 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Table 5. (continued).
⌬ log MIC (µmol/L)
Fragment
M. gypseum
T. mentagrophytes
120 h
72 h
120 h
72 h
3Ј,4Ј-Cl2
4Ј-Br
4Ј-F
–0.7208
–0.4014
–0.1003
–0.4203
–0.4012
–0.4203
–0.4203
0.3512
–1.1177
–0.5159
–0.301
–0.5159
–0.7524
–0.8173
–0.5159
0.1505
–1.03
–0.7295
–0.2006
–0.4282
–0.6646
–0.4282
–
–1.0448
–0.4431
–0.2007
–0.4431
–0.6687
–0.4431
–
3Ј-F
4Ј-CF3
4Ј-NO2
3Ј-NO2
4Ј-OCH3
µ0
–0.1004
1.6221
0.2341
1.637
1.313
1.7099
n = 23
n = 20
n = 22
n = 20
r = 0.9503
s = 0.2113
F = 6.45
r = 0.9273
s = 0.3218
F = 2.83
r = 0.9288
s = 0.3062
F = 4.71
r = 0.9399
s = 0.3121
F = 4.42
atives from the other groups. In all subseries studied, N-
benzylderivatives displayed better activities than N-phe-
nylderivatives.To evaluate the relationships between the
structure and antifungal activity against T. mentagro-
phytes and M. gypseum, the Free-Wilson method was
used (see Table 5).The antifungal activity of the deriva-
tives under study is relatively low, and only the activity
against the dermatophytic strains is significant.This pa-
per completed the results of our previous paper [3], and
the conclusions will be further utilized in our future work
towards the development of new antituberculotics.
Czech Republic), cyclohexane-acetone (3:1) was used as the
mobile phase.The 1H NMR and 13C NMR spectra of new com-
pounds were recorded in [D6]DMSO solutions at ambient tem-
perature on a Varian Mercury-Vx BB 300 spectrometer operat-
ing at 300 MHz for 1H, and 75 MHz for 13C.Chemical shifts were
recorded as δ values in parts per million (ppm) and were indi-
rectly referenced to tetramethylsilane via the solvent signal
1
(2.49 for H or 39.7 for 13C). The data are given in Table 2.
General Procedure for the Preparation of 3-hydoxypicolinani-
lides 2, N-benzyl-3-hydroxypicolinamides 3, 2-sulfanyl-
benzanilides 4 and, N-benzyl-2-sulfanylbenzamides 5:
a suspension of a substituted acid (0.02 mol) and a substituted
aniline or substituted benzylmides (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 solvents evaporated.The product was crys-
tallized from ethanol-water (yields in the range 75–90 %).
Acknowledgements
This work was supported by the research projects MSM
111600001, MSM 111600002, and grant No. 234/2000/
BCh/FaF from the Grant Agency of Charles University.
Microbiology
Antimycobacterial susceptibility testing. The following strains,
obtained from the Czech National Collection of Type Cultures
(CNCTC), National Institute of Public Health, Prague, were
used for the evaluation of in vitro antimycobacterial activity: M.
tuberculosis CNCTC My 331/8, M. kansasii CNCTC My 235/
80, and M. avium CNCTC My 330/88. The antimycobacterial
activity of the compounds was determined in the Sula semisyn-
thetic medium (SEVAC, Prague).This medium (with bovine se-
rum) is routinely used in the Czech Republic. Each strain was
simultaneously inoculated into a Petri dish containing the Lö-
wenstein-Jensen medium for the control of the sterility of the
inoculum and its growth. The compounds were added to the
medium in DMSO solutions. The final concentrations were
1000, 500, 250, 125, 62.5, 31, 16, 8, 4, 2 µmol/L. The MICs
were determined after incubation at 37 °C for 14 and 21 d (see
Table 3). MIC was the lowest concentration of an antimycobac-
terially effective substance (on the above concentration scale),
at which inhibition of the growth of the mycobacteria occurred.
Dedicated to Prof. Dr. Hans-Dietrich Stachel, München,
on the occasion of his 75th birthday.
Experimental
Chemistry
The melting points were determined on a Kofler apparatus.The
samples for analysis and antimycobacterial tests were dried
over P4O10 at 61 °C and 66 Pa for 24 h. Elemental analyses (C,
H, N) were performed on a CHNS-O CE elemental analyzer
(Fisions EA 1110, Milan) and were within 0.4 % of the theoreti-
cal values. The IR spectra were measured in KBr pellets on a
Nicolet Impact 400 apparatus; the wavenumbers are given in
cm–1.TLC was performed on silica gel plates precoated with a
fluorescent indicator Silufol UV 254 + 366 (Kavalier Votice,

Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Antimycobacterial and antifungal isosters of salicylamides 333
Table 6. Antifungalactivities(MIC, (µmol/L))of salicylanilides andtheirisosters against ninestrains ofpotentially patho-
genic fungi.
Ozn.
R
CA
CT
CK
CG
TB
Incubation time
24h
AF
AC
TM
MG
24h
48h
24h
48h
24h
48h
24h
48h
24h
48h
24h
48h
72h
120h
72h
120h
48h
1a
1b
1c
1d
1e
1f
H
4-CH3
4-Cl
125
250
250
500
250
500
500
500
250
500
125
250
250
250
62.5
62.5
31.25
62.5
62.5
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
31.25
62.5
31.25
62.5
15.63
62.5
62.5
>125
31.25
31.25
62.5
62.5
31.25
>125
7.81
31.25
31.25
31.25
3.91
7.81
7.81
7.81
3-Cl
31.25
>62.5
>62.5
>62.5
31.25
>62.5
>62.5
>62.5
>62.5
>62.5
31.25
>62.5
>62.5
>62.5
15.63
15.63
7.81
15.63
3,4-Cl2
4-Br
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
3.91
3.91
3.91
3.91
15.63
62.5
>125
>125
31.25
62.5
>125
>125
31.25
>125
62.5
>125
31.25
31.25
7.81
15.63
7.81
15.63
1g
1h
1i
4-F
62.5
125
125
>125
125
>125
125
125
125
>125
62.5
125
125
125
31.25
31.25
15.63
31.25
3-F
62.5
125
125
125
32.5
125
125
125
62.5
125
62.5
125
62.5
62.5
15.63
15.63
7.81
15.63
4-CF3
4-NO2
3-NO2
4-OCH3
H
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
31.25
>125
3.91
3.91
3.91
3.91
1j
31.25
62.5
>125
>125
62.5
62.5
62.5
125
31.25
>125
62.5
>125
62.5
62.5
15.63
15.63
7.81
7.81
1k
1l
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
7.81
15.63
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
62.5
125
62.5
125
2a
2b
2c
2d
2e
2f
125
250
500
500
250
250
125
250
500
500
250
500
250
250
125
250
62.5
125
4-CH3
4-Cl
125
125
250
>250
250
250
125
250
250
>250
250
250
125
125
125
125
62.5
62.5
62.5
>125
>125
>125
62.5
>125
>125
>125
>125
>125
>125
>125
125
>125
62.5
125
31.25
62.5
3-Cl
62.5
>125
>125
>125
>125
>125
62.5
>125
>125
>125
>125
>125
125
>125
62.5
>125
31.25
62.5
3,4-Cl2
4-Br
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
31.25
>250
2g
2i
4-F
62.5
125
250
>500
250
250
125
125
250
>500
125
125
125
125
125
125
62.5
62.5
4-CF3
4-NO2
125
250
500
500
250
500
62.5
125
500
500
125
250
125
125
125
125
62.5
62.5
2j
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250

334 Waisser et al.
Arch. Pharm. Pharm. Med. Chem. 2003, 336, 322–335
Table 6. (continued).
Ozn.
R
CA
CT
CK
CG
TB
Incubation time
24h
AF
AC
TM
MG
24h
48h
24h
48h
24h
48h
24h
48h
24h
48h
24h
48h
72h
120h
72h
120h
48h
2k
2l
3-NO2
4-OCH3
H
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
125
250
>500
>500
250
>500
125
250
>500
>500
250
>500
250
250
250
250
125
125
4a
4b
4c
4d
4e
4f
15.63
>125
125
>125
62.5
>125
>125
>125
15.63
>125
>125
>125
>125
>125
31.25
31.25
31.25
>125
4-CH3
4-Cl
7.81
15.63
31.25
31.25
31.25
>250
15.63
>250
15.63
31.25
>250
>250
>250
>250
31.25
62.53
1.25
31.25
125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
31.25
>125
3-Cl3
3,4-Cl2
4-Br
1.25
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>500
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>250
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
4g
4h
4l
4-F
7.81
>62.5
>62.5
>62.5
>62.5
>62.5
>62.5
>62.5
15.63
>62.5
>62.5
>62.5
>62.5
>62.5
15.63
31.25
>62.5
>62.5
3-F
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
>125
4-OCH3
7.81
>250
7.81
>250
31.25
>250
>250
>250
3.91
7.81
15.63
>250
>250
>250
7.81
>250
250
>250·
CA-Candida albicans ATCC 44859, CT-Candida tropicalis 156, CK-Candida krusei E28, CG-Candida glabrata 20/I,
TB-Trichosporon beigelii 1188, AF-Aspergillus fumigatus 231, AC-Absidia corymbifera 272,TM-Trichophyton menta-
grophytes 445, MG-Microsporum gypseum 27339.
Antifungal susceptibility testing. The compounds were dis-
solved in dimethylsulfoxide (DMSO). Nine fungal isolates (Tri-
chophyton mentagrophytes 445, Microsporum gypseum, As-
pergillus fumigatus 231, Absidia corymbifera 272, Candida al-
bicans ATTCC 44859 and Microsporum gypseum) from pa-
tients with suspected or proven mycosis were selected for in vit-
ro testing by the microdilution broth test. All strains were sub-
cultured on the Sabouraud dextrose agar (SDA) and main-
tained on the same medium at 4 °C.Prior to testing, each strain
was passaged onto SDA and fungal inocula were prepared by
suspending yeasts or spores in sterile 0.85 % saline. The cell
density was adjusted by means of the Bürker’s chamber to yield
a stock suspension of 5.0 × 105 CFU/mL. The final inoculum
was made by 1:20 dilution of the stock suspension with the test
medium. The in vitro antifungal activity of the compounds was
determined in the RPMI 1640 medium (Sevapharma, Prague)
buffered to pH 7.0 with 0.165 M morpholinopropansulfonic acid
(MOPS, Serva). The results are summarized in Table 6.
Calculations
All calculations were carried out with the use of the Multireg H
programme (Klemera) for Microsoft Excel.
References
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