Design, synthesis, and invitro antifungal activity of 1-[(4-substituted-benzyl)methylamino]-2-(2,4-difluorophenyl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ols

Article abstract of DOI:10.1002/cmdc.201000530

As part of our studies focused on the design of 1-[((hetero)aryl- and piperidinylmethyl)amino]-2-phenyl-3-(1H-1,2,4-triazol-1-yl)propan-2-ols as antifungal agents, we report the development of new extended benzylamine derivatives substituted at the para p

Full text of DOI:10.1002/cmdc.201000530

MED
DOI: 10.1002/cmdc.201000530
Design, Synthesis, and in vitro Antifungal Activity of 1-[(4-
Substituted-benzyl)methylamino]-2-(2,4-difluorophenyl)-3-
(1H-1,2,4-triazol-1-yl)propan-2-ols
Rꢀmi Guillon,[a] Fabrice Pagniez,[b] Francis Giraud,[a] Damien Crꢀpin,[a] Carine Picot,[b]
Marc Le Borgne,[a] Florent Morio,[b] Muriel Duflos,[a] Cꢀdric Logꢀ,*[a] and Patrice Le Pape[b]
As part of our studies focused on the design of 1-[((hetero)aryl-
and piperidinylmethyl)amino]-2-phenyl-3-(1H-1,2,4-triazol-1-yl)-
propan-2-ols as antifungal agents, we report the development
of new extended benzylamine derivatives substituted at the
para position by sulfonamide or retrosulfonamide groups
linked to alkyl or aryl chains. These molecules have broad-spec-
trum antifungal activities not only against Candida spp., includ-
ing fluconazole-resistant strains, but also against a filamentous
species (A. fumigatus). Concerning fluconazole resistance, se-
lected compounds exhibit the capacity to overcome CDR and
ERG11 gene upregulation and to maintain antifungal activity
despite a recognized critical CYP51 substitution in C. albicans
isolates. Synthesis, investigation of the mechanism of action by
sterol analysis in a C. albicans strain, and structure–activity rela-
tionships (SARs) are reported.
Introduction
During the last few decades, fungi have become a major
threat to many hospitalized patients particularly for those who
are severely immunocompromised and are therefore highly
susceptible to systemic fungal infections, most often caused
by Candida and Aspergillus species.[1] Currently, four main
chemical families (having four global cellular targets in fungal
cells) are used for the treatment of invasive fungal infections:
polyenes, pyrimidine analogues, azoles, and echinocandins.
Among them, azoles are the largest class of antifungal agents
in clinical use. They mainly target P450-dependent sterol 14a-
demethylase (CYP51, Erg11), encoded by the ERG11 gene, a
key enzyme in fungal ergosterol biosynthesis.[2] This inhibition
leads to depletion of ergosterol in the cell membrane and ac-
cumulation of toxic intermediate sterols, causing increased
membrane permeability and inhibition of fungal growth. The
‘first-generation’ triazoles (fluconazole and itraconazole) con-
siderably improved the treatment of some serious fungal infec-
tions, such as candidiasis. However, these earlier agents have
limitations related to their narrow-spectrum antifungal activi-
ty.[3] The lack of activity against filamentous fungi and emer-
gence of acquired resistance of some Candida isolates repre-
sent well-known limits of fluconazole, whereas itraconazole
has an unpredictable absorption in oral formulations and con-
siderable metabolic variability.[4] Currently, among the ‘second-
generation’ triazoles, including voriconazole,[5] posaconazole,[6]
ravuconazole,[7] isavuconazole,[8] and albaconazole,[9] only vori-
conazole and posaconazole are available for clinical manage-
ment of invasive fungal infections. Both compounds have a
very broad-spectrum of antifungal activities that includes Can-
dida species, filamentous, and dimorphic fungi. Ravuconazole,
isavuconazole, and albaconazole are currently in various stages
of development and have shown very potent in vitro activity
against species of Candida, Cryptococcus, and Aspergillus.[10]
However, despite the arrival of these new effective drugs,
some therapeutic problems remain, in particular emerging
fungal species, variable drug bioavailability, some toxicity, lack
of either oral or intravenous preparations, significant drug in-
teractions for some agents, and development of resistance.[11]
Antifungal resistance is a serious concern due to the limited
number of available agents. In Candida species, four major
mechanisms of resistance to azoles have been described:
1) decreased intracellular accumulation of azoles due to the
overexpression of genes encoding efflux transporters belong-
ing to the ATP-binding cassette (ABC) superfamily (CDR1 and
CDR2) or major facilitator superfamily (MDR1);[12,13] 2) genetic
alterations in the ERG11 gene leading to amino acid substitu-
tions in the target enzyme CYP51 that decrease drug bind-
ing;[14–16] 3) alterations in the sterol biosynthesis pathway that
bypass the accumulation of toxic sterols;[17,18] and 4) overex-
pression of the ERG11 gene.[19] Therefore, the antifungal agent
is overwhelmed, and routine therapeutic concentrations can
no longer effectively inhibit ergosterol synthesis.[20]
To date, more than 140 different amino acid substitutions
have been detected in CYP51 of clinical isolates of C. albi-
[a] Dr. R. Guillon, Dr. F. Giraud, D. Crꢀpin, Prof. M. Le Borgne, Prof. M. Duflos,
Dr. C. Logꢀ
Universitꢀ de Nantes, Nantes Atlantique Universitꢀs, Dꢀpartement de
Pharmacochimie, Cibles et Mꢀdicaments des Infections, de l’Immunitꢀ et du
Cancer, IICIMED-EA 1155, UFR Sciences Pharmaceutiques, 1 rue Gaston Veil,
44035 Nantes Cedex 1 (France)
Fax: (+33) 240-412-876
E-mail: cedric.loge@univ-nantes.fr
[b] Dr. F. Pagniez, C. Picot, F. Morio, Prof. P. Le Pape
Universitꢀ de Nantes, Nantes Atlantique Universitꢀs, Dꢀpartement de
Parasitologie et Mycologie Mꢀdicale, Cibles et Mꢀdicaments des Infections,
de l’Immunitꢀ et du Cancer, IICIMED-EA 1155, UFR Sciences
Pharmaceutiques, 1 rue Gaston Veil, 44035 Nantes Cedex 1 (France)
816
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ChemMedChem 2011, 6, 816 – 825

Antifungal Agents
As part of our research project
focused on the optimization of
azole antifungal agents based
on the modification of the side
chain attached to the pharmaco-
phore, we recently reported on
the design and synthesis of 1-
[((hetero)aryl- or piperidinylme-
thyl)amino]-2-phenyl-3-(1H-1,2,4-
triazol-1-yl)propan-2-ols showing
various degrees of antifungal ac-
tivity against Candida albicans
and
Aspergillus
fumigatus
strains.[34–37] The benzylamine
series bearing H-bond acceptor
entities in the para position of
the benzyl group and
a N-
methyl group in the linker gave
the most active compounds both in C. albicans and in A. fumi-
gatus strains, in accordance with the receptor-based pharma-
cophore model published by Sheng et al.[29]
cans.[21] Most of these substitutions, instead of being randomly
dispersed, are clustered into three hot spot regions ranging
from amino acids 105 to 165, 266 to 287, and 405 to 488.[22]
The contribution of each individual mutation to azole resist-
ance is, however, difficult to estimate because ERG11 mutations
often occur in combination (between two and four combined
mutations) in the same allele and because resistance mecha-
nisms are often combined in azole-resistant C. albicans iso-
lates.[18,22,23]
Herein, we report the development of new benzylamine de-
rivatives substituted in the para position by sulfonamide (I) or
retrosulfonamide groups (II) linked to alkyl or aryl chains.
The long-awaited structural information of CYP51 from
pathogenic fungi could certainly accelerate the discovery of
novel antifungal agents as well as provide insight into a better
understanding of the resistance mechanisms. In the absence of
such crystal structures, several three-dimensional models of
CYP51 from C. albicans, A. fumigatus, or Cryptococcus neofor-
mans have been reported[24–27] on the basis of a prokaryotic
CYP51 from M. tuberculosis[28] and interactions with azoles have
been explored by flexible molecular docking. In particular, the
receptor-based pharmacophore model published by Sheng
et al. was used to guide the rational optimization of the azole
antifungal agents and highlighted the importance of Tyr118
and Ser378 residues of C. albicans sterol 14a-demethylase
P450 (CACYP51) in the stabilization of inhibitors.[29] Tyr118, a
highly conserved residue in the CYP51 family, may be a poten-
These structures are easily prepared, retained all the main char-
acteristics mentioned above, and may be of sufficient size to
avoid mutations near the heme site. The synthesis and biologi-
cal in vitro evaluation against various Candida spp. and A. fumi-
gatus strains are described.
tial site for the p–p interaction with the inhibitor, whereas Results and Discussion
Ser378, a conserved amino acid across the fungal CYP51 en-
Chemistry
zymes, could form a specific hydrogen bonding interaction.
The role of the Tyr118 residue in determining azole susceptibil-
ity was also validated by site-directed mutagenesis.[30,31] More-
over, to combat infection more efficiently, one strategy is to in-
crease the size of the inhibitor to cover a larger area within
the active site of fungal CYP51.[32,33] In particular, Xiao et al.
provided a model for the binding of azoles with extended side
chains in CYP51 from C. albicans and A. fumigatus. Specifically,
itraconazole and posaconazole appear to be less affected by
mutations near the heme site than either fluconazole or vori-
conazole and this could be explained by tighter affinity and/or
compensatory adjustments within the active site.[24]
The synthetic route of target compounds 12–20 is outlined in
Scheme 1. These molecules were obtained by nucleophilic sub-
stitution of previously described[36] key intermediate 11 with
appropriate substituted 4-(bromomethyl)benzenesulfonamides
prepared by the reaction of commercially available 4-(bromo-
methyl)benzenesulfonylchloride 1 with amines. Compound 2
was obtained following a synthetic procedure reported in the
literature by the action of aqueous ammonia in THF.[38] Sulfona-
mides 3–6 and 7–10 were respectively synthesized in good
yields by reaction of 1 with alkylamines in the presence of
Hꢂnig’s base in THF or with anilines in dichloromethane and
pyridine. The nucleophilic substitution reaction by the amino
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817

C. Logꢀ et al.
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Antifungal activity
The in vitro antifungal activities
of all compounds were evaluat-
ed against human pathogenic
fungi
(C. albicans,
C. krusei,
C. glabrata, C. parapsilosis, and
A. fumigatus) and are summar-
ized in Table 1. The MIC values
(in mgmLÀ1) of fluconazole, vori-
conazole, and itraconazole are
also shown as reference.
With the C. albicans CA98001
strain, all compounds displayed
a high level of activity, with MIC
values ranging from 1.0 to
69.0 ngmLÀ1, similar to that of
voriconazole or fluconazole. For
the A. fumigatus AF98003 strain,
biological results are promising
Scheme 1. Reagents and conditions: a) NH4OH, THF, RT, 1 h, 98%; b) alkylamines, N,N-diisopropylethylamine, THF,
RT, 1 h, 70–97%; c) anilines, pyridine, CH2Cl2, 08C, 2 h, 81–90%; d) 2–10, N,N-diisopropylethylamine, CH3CN, RT,
24 h, 61–85%.
alcohol 11 on derivatives 2–10 in the presence of Hꢂnig’s base
in acetonitrile afforded nine original compounds (12–20).
Scheme 2 outlines the synthesis of compounds 23–26 start-
ing from the key intermediate 11. Reaction with 4-nitrobenzyl-
bromide using Hꢂnig’s base in acetonitrile gave compound 21
with MIC values mostly ranging from 767.0 to 3570.0 ngmLÀ1
,
only two- to eightfold greater than that of itraconazole. On
both of these strains, compounds bearing the most extreme
side chain from a steric point of view (12 compared with 19 or
20) share the same antifungal values, and comparison of sulfo-
namides 15, 17, 19, and 20 with their retrosulfonamide ana-
logues 23, 24, 25, and 26, did not reveal significant differences
in activity, except for compound 23 (the sole representative in
the alkyl series) on the A. fumigatus strain.
A significant broad-spectrum antifungal activity was also ob-
served for some molecules (15–20 and 25) against C. krusei,
C. glabrata, and C. parapsilosis strains confirming their potential
in targeting fluconazole low-susceptible strains and flucona-
zole-intrinsically resistant Candida species.
Sterol analysis
The mechanism of action of this series of azoles was investigat-
ed by studying inhibition of C. albicans CA98001 ergosterol
biosynthesis after treatment by 17, one of the most active
compounds. As shown in Table 2, this molecule strongly inhib-
ited ergosterol biosynthesis while lanosterol accumulated.
About 60% of ergosterol biosynthesis inhibition was obtained
with a concentration of 0.020 mgmLÀ1, near the MIC value
(4.0 ngmLÀ1). At a higher concentration (0.1 mgmLÀ1), this
effect was maximal showing that inhibition of ergosterol bio-
synthesis by this compound is dose-dependent. Also, methylat-
ed sterols at the C14 position also accumulated (eburicol, 14-
methylfecosterol and 14-methylepisterol) in a dose-dependent
manner. Accumulation of lanosterol and production of 14-me-
thylated sterols are typically a result of a 14a-demethylase
blockage by triazole derivatives. However, the low proportion
of 14-methyl-3,6-diol could also be explained by further inhibi-
tion of D5,6-desaturase encoded by the ERG3 gene.[41]
Scheme 2. Reagents and conditions: a) 4-nitrobenzylbromide, N,N-diisopro-
pylethylamine, CH3CN, RT, 24 h, 77%; b) 5% Pd/C, H2, EtOH, RT, 5 bar, 2 h,
72%; c) isopropylsulfonyl chloride, pyridine, RT, 2 h, 47%; d) benzenesulfonyl
chlorides, In, CH3CN, RT, 15 h, 29–68%.
in 77% yield. Catalytic hydrogenation of the nitro aromatic
group using 5% Pd/C in ethanol afforded the corresponding
amine 22, which was condensed with isopropylsulfonyl chlo-
ride in pyridine to give molecule 23.[39] Targeted aromatic com-
pounds 24–26 were prepared by indium catalysis in acetoni-
trile with benzenesulfonyl chlorides.[40]
818
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Antifungal Agents
Table 1. In vitro antifungal activities of compounds 12–20 and 23–26 against Candida and A. fumigatus strains.
[a]
Compd
Y
MIC [mgmLÀ1
]
CA98001
CK506
CK8
CG468
CAPA1
CAPA2
AF98003
12
13
14
15
16
17
18
19
20
23
24
25
26
-SO2NH2
-SO2-morpholine
-SO2-NH-Pr
-SO2-NH-iPr
-SO2-NH-iBu
-SO2-NH-Ph
-SO2-NH-Ph-(p)Pr
-SO2-NH-Ph-(p)iPr
-SO2-NH-Ph-(p)NO2
-NH-SO2-iPr
-NH-SO2-Ph
-NH-SO2-Ph-(p)iPr
-NH-SO2-Ph-(p)NO2
–
0.013Æ0.001
0.018Æ0.001
13.0Æ0.4 13.2Æ1.1
3.4Æ1.6
0.91Æ0.12
0.88Æ0.17
0.91Æ0.07
0.58Æ0.01
0.038Æ0.030
0.10Æ0.02
0.12Æ0.02
0.10Æ0.01
0.017Æ0.005
4.5Æ1.4
0.75Æ0.16
1.1Æ0.3
2.45Æ0.04
3.25Æ0.10
0.767Æ0.24
2.35Æ0.05
1.88Æ0.20
3.08Æ0.10
1.56Æ0.44
3.33Æ0.06
2.57Æ0.11
29.0Æ0.50
3.29Æ0.05
2.89Æ0.06
3.57Æ0.06
–
13.4Æ0.7
12.6Æ0.4
5.7Æ0.1
2.2Æ0.7
1.6Æ0.1
1.2Æ0.1
1.8Æ0.1
3.5Æ0.7
2.7Æ1.0
1.8Æ0.1
1.8Æ0.36
1.0Æ0.3
1.8Æ0.1
1.8Æ0.1
0.021Æ0.002
1.57Æ0.03
0.014Æ0.001
<0.001
1.5Æ0.5
0.002Æ0.001
<0.001
0.76Æ0.01
0.84Æ0.2
0.12Æ0.06
0.23Æ0.05
0.62Æ0.06
0.73Æ0.16
3.1Æ0.9
<0.001
0.004Æ0.001
0.069Æ0.051
0.030Æ0.003
0.018Æ0.002
0.026Æ0.008
0.015Æ0.004
0.023Æ0.001
0.030Æ0.008
2.0Æ1.0
1.6Æ0.03
0.85Æ0.22 <0.001
1.9Æ0.2
<0.001
14.6Æ0.3
1.40Æ0.03
1.9Æ0.1
18.3Æ0.2
7.7Æ0.1
0.45Æ0.04
–
17.5Æ0.2 15.1Æ0.6
10.6Æ1.9
2.0Æ0.1
1.7Æ0.2
1.5Æ0.1
1.2Æ0.2
0.58Æ0.10
0.18Æ0.01
1.7Æ0.2
0.15Æ0.03
1.91Æ0.36
15.3Æ1.6 18.3Æ2.0
fluconazole
voriconazole
itraconazole
0.036Æ0.021 >30
12.9Æ0.9
0.24Æ0.07
–
>30
>30
–
–
0.005Æ0.001
1.3Æ0.3
0.95Æ0.13
0.36Æ0.01
0.15Æ0.01
0.42Æ0.04
–
–
–
–
[a] Values represent the mean ÆSD of experiments performed in triplicate; C. albicans (CA98001), C. krusei (CK506, CK8), C. glabrata (CG468), C. parapsilosis
(CAPA1, CAPA2), and A. fumigatus (AF98003).
Table 2. Effect of compound 17 on the sterol composition of C. albicans
CA98001.
Table 3. In vitro antifungal activities of compounds 17–19 and 25 against
C. albicans strains with reduced fluconazole susceptibility.
Sterols[a]
Control
17 [mgmLÀ1
0.020
]
0.1
lanosterol
eburicol
zymosterol
ergosta-5,8-dienol
episterol
14-methylfecosterol
14-methylepisterol
14-methyl-3,6-diol
ergosterol
0.5
0.6
0.3
1.3
0.3
–
–
–
97.0
22.5
31.0
0.9
2.6
0.7
2.1
3.2
0.4
36.7
38.3
44.2
–
–
–
5.7
9.4
2.4
0.0
[a]
Compd
Y
MIC [mgmLÀ1
]
DSY735
CAAL-74
17
18
19
-SO2-NH-Ph
-SO2-NH-Ph-(p)Pr
-SO2-NH-Ph-(p)iPr
-NH-SO2-Ph-(p)iPr
–
1.03Æ0.26
0.29Æ0.01
1.05Æ0.31
0.24Æ0.01
12.55
0.06Æ0.01
0.011Æ0.001
0.17Æ0.01
0.12Æ0.03
[a] Sterols of interest were identified by their mass spectra; the area
under the curve (AUC) of each peak was used to calculate the ratio of
(sterol AUC)/(sum of sterols AUC).
25
fluconazole
>30
[a] Values represent the mean ÆSD of experiments performed in tripli-
cate.
Biological evaluation against C. albicans isolates with
decreased fluconazole susceptibility
To evaluate more precisely the biological interest of these
compounds, we investigated their potency against two C. albi-
cans strains having identified mechanisms of resistance. These
include TAC1 (transcriptional activator of CDR genes) mutation
and TAC1 and ERG11 upregulation after formation of isochro-
mosome 5L (DSY735) and amino acid substitutions Tyr132Phe/
Gly448Val (CAAL-74). Compounds 17–19 and 25 were selected
because of their broad-spectrum antifungal activity and rela-
tively long side chain positioned in the benzylamine linker. The
MIC values (in mgmLÀ1) are presented in Table 3.
Interestingly, compared to fluconazole, all these extended
compounds exhibited high antifungal activities against the se-
lected strains, showing that overexpression of efflux pumps
and ERG11 or specific point substitutions in the CYP51 enzyme,
which result in fluconazole resistance, could be rationally over-
come.
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C. Logꢀ et al.
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Molecular modeling
Candida spp. including fluconazole-resistant strains but also
against a filamentous species (A. fumigatus), which are the
most prevalent fungal pathogens. Concerning fluconazole re-
sistance, selected compounds demonstrate a capacity to over-
come CDR and ERG11 gene upregulation and to maintain anti-
fungal activity despite a recognized critical CYP51 substitution
in C. albicans isolates. Additional investigations including
emerging molds (Scedosporium spp., Fusarium spp., and Zygo-
mycetes), a large number of azole-resistant clinical isolates, in
vitro evaluation on recombinant CYP51 from C. albicans and
toxicity studies could be undertaken to check the usefulness of
these compounds for further optimization.
Despite a mechanism of action that could imply both inhibi-
tion of 14a-demethylase (CYP51) and D5,6-desaturase, we per-
formed the docking of compound 17 in our model of C. albi-
cans CYP51.[37,42] According to the receptor-based pharmaco-
phore model,[29] the putative binding solution suggests key hy-
drogen bonding interactions between the sulfonyl group and
both Ser378 and His377 residues (Figure 1). On the C. albicans
Experimental Section
Chemistry
General methods: Melting points were determined using an Elec-
trothermal IA9300 digital melting point apparatus and are reported
uncorrected. 1H and 13C NMR spectra were recorded on a Bruker
AC250 (250 MHz) or on Bruker Avance 400 spectrometer
(400 MHz). Chemical shifts are expressed as d values (ppm) relative
to tetramethylsilane as internal standard (in NMR description, s=
singlet, d=doublet, t=triplet, q=quadruplet, sext=sextuplet,
m=multiplet, and b=broad). Coupling constants J are given in
Hertz. IR spectra were obtained in KBr pellets using a PerkinElmer
Paragon FTIR 1000 PC spectrometer. Only the most significant ab-
sorption bands have been reported. Electrospray mass spectromet-
ric analysis was performed on a Waters Acquity UPLC System ZQ
2000 single quadrupole. All tested compounds displayed more
than 95% purity. All reactions were monitored by thin-layer chro-
matography (TLC) using 0.2 mm-thick silica gel plates 60F-254
(5735 Merck). Column chromatography was carried out using silica
gel 60 (70–230 Mesh, ASTM, Merck). Chemicals and solvents used
are commercially available.
Figure 1. Proposed binding solution of compound (S)-17 in the putative
active site pocket (channel 2) of C. albicans CYP51. Hip377 is the protonated
form of the histidine residue. The backbone is shown in tube representation
(Sybyl v. 8.0, Tripos Associates Inc., St. Louis, MO, USA).
4-(Bromomethyl)benzenesulfonamide (2): NH4OH (28% in H2O)
(0.93 mL, 13.83 mmol) was added to a stirred solution of 4-bromo-
CAAL-74 strain, substitutions of Tyr132Phe is known to confer
azole resistance.[43] Tyr132 is located on the enzyme’s flexible
BC loop, which adopts the open conformation observed in the
crystal structure of CYP51 from M. tuberculosis which is used as
a template. This conformation positions this residue away from
the active site. However, we can assume that, given the recent
crystal structures of CYP51 from human and Trypanosomatidae
in complex with azole antifungal inhibitors,[44,45] this residue
could come into close proximity with the active site as the BC
loop adapts a closed conformation. Thus, the activity observed
for the larger compound 17 relative to fluconazole could be
explained by tighter affinity and/or compensatory adjustments
within the active site. The involvement of the Gly448Val substi-
tution is more difficult to explain because this amino acid is
only described in combination with Tyr132 in clinical isola-
tes,[21,43b] and is part of a large specific loop insertion, located
far from the active site, not modeled in detail.
methylbenzenesulfonyl chloride
1 (1.0 g, 3.71 mmol) in THF
(12 mL) at 08C. The solution was stirred at RT for 1 h. the mixture
was diluted with H2O and the product was extracted with EtOAc.
Organic layers were dried over anhydrous Na2SO4 and concentrat-
ed in vacuo to give compound 2 as a white powder (910 mg,
98%): Rf =0.40 (CH2Cl2/EtOH 19:1); mp: 176–1778C; 1H NMR
(400 MHz, [D6]DMSO): d=4.80 (s, 2H), 7.43 (s, 2H), 7.67 (d, 2H, J=
8.2 Hz), 7.84 ppm (d, 2H, J=8.2 Hz); IR (KBr): n˜ =3361, 2957, 2929,
1569, 1507, 1463, 1329, 1153, 673 cmÀ1; MS (ESI) m/z (%): 78.9 (71),
80.9 (100), 248.0 (58), 250.0 (71) [MÀH]+.
4-{[4-(Bromomethyl)phenyl]sulfonyl}morpholine (3): N,N-diisopro-
pylethylamine (154 mL, 0.89 mmol) was added to a stirred solution
of morpholine (71 mL, 0.82 mmol) in THF (3 mL) at 08C followed by
dropwise addition of 4-bromomethylbenzenesulfonyl chloride 1
(200 mg, 0.74 mmol) in THF (4 mL). The solution was stirred at RT
for 1 h. The mixture was diluted with H2O and the product was ex-
tracted with CH2Cl2. Organic layers were dried over anhydrous
Na2SO4 and concentrated in vacuo. The residue was purified with
silica gel column chromatography (CH2Cl2) to yield compound 3 as
a white powder (166 mg, 70%): Rf =0.40 (CH2Cl2); mp: 147–1488C;
1H NMR (400 MHz, [D6]DMSO): d=2.90 (t, 4H, J=4.6 Hz), 3.66 (t,
4H, J=4.6 Hz), 4.84 (s, 2H), 7.77 ppm (s, 4H); IR (KBr): n˜ =1339,
1164, 1099, 940, 729 cmÀ1; MS (ESI) m/z (%): 82.8 (100), 319.8 (6),
321.9 (6) [M+H]+.
Conclusions
In summary, we report on the synthesis and biological evalua-
tion of novel triazole derivatives. These extended molecules
have broad-spectrum antifungal activities not only against
820
www.chemmedchem.org
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
ChemMedChem 2011, 6, 816 – 825

Antifungal Agents
4-(Bromomethyl)-N-(propyl)benzenesulfonamide (4): Using the
synthetic procedure used for compound 3 starting from 4-bromo-
methylbenzenesulfonyl chloride 1 (300 mg, 1.11 mmol) and n-pro-
pylamine (183 mL, 2.22 mmol) to yield compound 4 as a light
brown powder (265 mg, 82%): Rf =0.35 (CH2Cl2); mp: 71–728C;
1H NMR (400 MHz, [D6]DMSO): d=0.82 (t, 3H, J=7.4 Hz), 1.40
(sext, 2H, J=7.4 Hz), 2.71 (m, 2H), 4.80 (s, 2H), 7.65 (t, 1H, J=
6.0 Hz), 7.69 (d, 2H, J=8.4 Hz), 7.80 ppm (d, 2H, J=8.4 Hz); IR
(KBr): n˜ =3266, 2965, 2878, 1323, 1153, 669 cmÀ1; MS (ESI) m/z (%):
82.9 (100), 291.9 (8), 293.9 (8) [M+H]+.
2.81 (m, 1H), 4.75 (s, 2H), 7.03 (d, 2H, J=7.5 Hz), 7.13 (d, 2H, J=
7.5 Hz), 7.63 (d, 2H, J=7.5 Hz), 7.77 (d, 2H, J=7.5 Hz), 10.24 ppm
(s, 1H); IR (KBr): n˜ =3268, 2963, 1510, 1446, 1326, 1161, 683 cmÀ1
MS (ESI) m/z (%): 82.8 (100), 367.9 (4), 369.9 (4) [M+H]+.
;
4-(Bromomethyl)-N-(4-nitrophenyl)benzenesulfonamide
(10):
Using the synthetic procedure used for compound 7 starting from
4-bromomethylbenzenesulfonyl chloride 1 (200 mg, 0.74 mmol)
and 4-nitroaniline (93 mg, 0.67 mmol) to yield crude product 10 as
a light brown powder (200 mg); MS (ESI) m/z (%): 369.0 (96), 371.0
(100) [MÀH]+.
4-(Bromomethyl)-N-(isopropyl)benzenesulfonamide (5): Using
the synthetic procedure used for compound 3 starting from 4-bro-
momethylbenzenesulfonyl chloride 1 (200 mg, 0.74 mmol) and iso-
propylamine (127 mL, 1.48 mmol) to yield compound 5 as a white
powder (210 mg, 97%): Rf =0.40 (CH2Cl2); mp: 95–968C; 1H NMR
(400 MHz, [D6]DMSO): d=0.97 (d, 6H, J=6.7 Hz), 3.29 (sep, 1H, J=
6.7 Hz), 4.80 (s, 2H), 7.67 (m, 3H), 7.82 ppm (dd, 2H, J=6.4 Hz, J=
1.8 Hz); IR (KBr): n˜ =3299, 1420, 1312, 1253, 676 cmÀ1; MS (ESI) m/z
(%): 82.9 (100), 291.9 (2), 293.9 (2) [M+H]+.
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)benzenesulfonamide (12): N,N-diisopro-
pylethylamine (114 mL, 0.66 mmol) was added to a stirred solution
of 11 (161 mg, 0.60 mmol) in CH3CN (5 mL) followed by the addi-
tion of compound 2 (150 mg, 0.60 mmol). The solution was stirred
at RT for 24 h. The solvent was removed under reduced pressure
and the residue was partitioned between CH2Cl2 and H2O. Organic
layers were dried over anhydrous Na2SO4 and concentrated in va-
cuo. The residue was purified using silica gel column chromatogra-
phy (CH2Cl2 and CH2Cl2/EtOH 99:1) to yield compound 12 as a
white powder (215 mg, 82%): Rf =0.75 (CH2Cl2/EtOH 95:5); mp:
4-(Bromomethyl)-N-(isobutyl)benzenesulfonamide (6): Using the
synthetic procedure used for compound 3 starting from 4-bromo-
methylbenzenesulfonyl chloride 1 (300 mg, 1.11 mmol) and isobu-
tylamine (221 mL, 2.22 mmol) to yield compound 6 as a white
powder (300 mg, 88%): Rf =0.50 (CH2Cl2); mp: 98–998C; 1H NMR
(400 MHz, [D6]DMSO): d=0.84 (d, 6H, J=6.7 Hz), 1.65 (m, 1H, J=
6.7 Hz), 2.59 (d, 2H, J=6.7 Hz), 4.80 (s, 2H), 7.68 (m, 3H), 7.81 ppm
(d, 2H, J=6.4 Hz); IR (KBr): n˜ =3265, 2959, 1416, 1320, 1149,
674 cmÀ1; MS (ESI) m/z (%): 82.9 (100), 305.9 (10), 308.0 (12)
[M+H]+.
1
151–1528C; H NMR (400 MHz, [D6]DMSO): d=2.09 (s, 3H), 2.79 (d,
1H, J=14.6 Hz), 3.10 (d, 1H, J=14.6 Hz), 3.47 (d, 1H, J=14.0 Hz),
3.70 (d, 1H, J=14.0 Hz), 4.57 (s, 2H), 5.83 (s, 1H), 7.01 (ddd, 1H,
J
J
J
H–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.21 (ddd, 1H, JH–F =J’H–F =9.2 Hz,
H–H =2.4 Hz), 7.24 (d, 2H, J=8.3 Hz), 7.32 (s, 2H), 7.46 (ddd, 1H,
H–H =8.4 Hz,
JH–F =J’H–F =6.8 Hz), 7.71 (d, 2H, J=8.3 Hz), 7.80
(s, 1H), 8.33 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=43.7,
56.1, 62.0, 63.3, 75.5, 103.9, 110.8, 125.6 (2C), 126.2, 128.0 (2C),
130.0, 142.8, 143.5, 145.1, 150.7 ppm, (CF not visible); IR (KBr):
n˜ =3312, 1612, 1499, 1417, 1336, 1155 cmÀ1; MS (ESI) m/z (%):
438.0 (100) [M+H]+; UPLC purity 96%.
4-(Bromomethyl)-N-(phenyl)benzenesulfonamide (7): Pyridine
(60 mL, 0.74 mmol) was added to a stirred solution of aniline
(62 mL, 0.67 mmol) in CH2Cl2 (5 mL) at 08C followed by the drop-
wise addition of 4-bromomethylbenzenesulfonyl chloride
1
2-(2,4-Difluorophenyl)-1-{methyl[4-(morpholinosulfonylphenyl)-
methyl]amino}-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (13): Using the
synthetic procedure used for compound 12 starting from 11
(100 mg, 0.37 mmol) and 3 (119 mg, 0.37 mmol) to yield com-
pound 13 as a white powder (160 mg, 85%): Rf =0.15 (CH2Cl2/EtOH
98:2); mp: 57–588C; 1H NMR (400 MHz, [D6]DMSO): d=2.15 (s,
3H), 2.79 (d, 1H, J=13.7 Hz), 2.86 (m, 4H), 3.06 (d, 1H, J=13.7 Hz),
3.55 (d, 1H, J=14.0 Hz), 3.66 (m, 4H), 3.73 (d, 1H, J=14.0 Hz), 4.52
(d, 1H, J=14.3 Hz), 4.61 (d, 1H, J=14.3 Hz), 5.84 (s, 1H), 7.00 (ddd,
(200 mg, 0.74 mmol) in CH2Cl2 (3 mL). The solution was stirred at
08C for 2 h. The mixture was diluted with CH2Cl2 and washed with
5m HCl. Organic layers were dried over anhydrous Na2SO4 and con-
centrated in vacuo. The residue was purified on silica gel column
chromatography (CH2Cl2) to yield compound 7 as a white powder
(185 mg, 84%): Rf =0.35 (CH2Cl2); mp: 124–1258C; 1H NMR
(400 MHz, [D6]DMSO): d=4.74 (s, 2H), 7.06 (t, 1H, J=7.2 Hz), 7.12
(d, 2H, J=7.2 Hz), 7.26 (t, 2H, J=7.2 Hz), 7.63 (d, 2H, J=8.4 Hz),
7.77 (d, 2H, J=8.4 Hz), 10.36 ppm (s, 1H); IR (KBr): n˜ =3232, 1594,
1484, 1338, 1163, 698 cmÀ1; MS (ESI) m/z (%): 78.9 (26), 80.8 (25),
324.0 (94), 326.0 (100) [MÀH]+.
1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.17 (ddd, 1H, JH–F =J’H–F
=
9.2 Hz, JH–H =2.4 Hz), 7.44 (m, 3H), 7.64 (d, 2H, J=8.3 Hz), 7.78 (s,
1H), 8.32 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=44.1,
46.1 (2C), 56.1, 62.2, 63.3, 65.5 (2C), 75.5, 103.9, 110.9, 126.3, 127.7
(2C), 129.5 (2C), 130.1, 132.9, 145.1, 145.5, 150.7 ppm, (CF not visi-
ble); IR (KBr): n˜ =3423, 1615, 1503, 1348, 1270, 1167, 1102 cmÀ1; MS
(ESI) m/z (%): 508.0 (100) [M+H]+; UPLC purity 97%.
4-(Bromomethyl)-N-(4-propylphenyl)benzenesulfonamide
(8):
Using the synthetic procedure used for compound 7 starting from
4-bromomethylbenzenesulfonyl chloride 1 (200 mg, 0.74 mmol)
and 4-propylaniline (100 mL, 0.67 mmol) to yield compound 8 as a
light red powder (225 mg, 90%): Rf =0.60 (CH2Cl2); mp: 85–868C;
1H NMR (400 MHz, [D6]DMSO): d=0.86 (t, 3H, J=7.3 Hz), 1.52
(sext, 2H, J=7.3 Hz), 2.46 (t, 2H, J=7.3 Hz), 4.74 (s, 2H), 7.01 (d,
2H, J=8.4 Hz), 7.07 (d, 2H, J=8.4 Hz), 7.62 (d, 2H, J=8.3 Hz), 7.74
(d, 2H, J=8.3 Hz), 10.21 ppm (s, 1H); IR (KBr): n˜ =3274, 2928, 2860,
1508, 1387, 1328, 1159, 686 cmÀ1; MS (ESI) m/z (%): 78.8 (28), 80.9
(26), 366.1 (100), 368.1 (87) [MÀH]+.
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)-N-(propyl)benzenesulfonamide
(14):
Using the synthetic procedure used for compound 12 starting
from 11 (138 mg, 0.51 mmol) and 4 (150 mg, 0.51 mmol) to yield
compound 14 as a light brown powder (150 mg, 61%): Rf =0.40
(CH2Cl2/EtOH 95:5); mp: 52–538C; 1H NMR (400 MHz, [D6]DMSO):
d=0.81 (t, 3H, J=7.2 Hz), 1.38 (m, 2H), 2.13 (s, 3H), 2.69 (q, 2H,
J=7.2 Hz), 2.80 (d, 1H, J=13.6 Hz), 3.07 (d, 1H, J=13.6 Hz), 3.50
(d, 1H, J=14.0 Hz), 3.69 (d, 1H, J=14.0 Hz), 4.53 (d, 1H, J=
4-(Bromomethyl)-N-(4-isopropylphenyl)benzenesulfonamide (9):
Using the synthetic procedure used for compound 7 starting from
4-bromomethylbenzenesulfonyl chloride 1 (200 mg, 0.74 mmol)
and 4-isopropylaniline (92 mL, 0.67 mmol) to yield compound 9 as
a light red powder (205 mg, 81%): Rf =0.45 (CH2Cl2); mp: 100–
1018C; 1H NMR (400 MHz, [D6]DMSO): d=1.14 (d, 6H, J=6.4 Hz),
14.4 Hz), 4.59 (d, 1H, J=14.4 Hz), 5.83 (s, 1H), 7.00 (ddd, 1H, JH–F
H–H =8.4 Hz, JH–H =2.4 Hz), 7.18 (ddd, 1H, JH–F =J’H–F =9.2 Hz, JH–H
2.4 Hz), 7.29 (d, 2H, J=7.6 Hz), 7.45 (ddd, 1H, JH–H =8.4 Hz, JH–F
=
=
=
J
J’H–F =6.8 Hz), 7.54 (m, 1H), 7.68 (d, 2H, J=7.6 Hz), 7.79 (s, 1H),
ChemMedChem 2011, 6, 816 – 825
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemmedchem.org
821

C. Logꢀ et al.
MED
8.33 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=11.3, 22.5,
43.9, 44.5, 56.0, 62.1, 63.2, 75.5, 103.9, 110.8, 126.2, 126.4 (2C),
129.1 (2C), 130.0, 139.2, 144.1, 145.1, 150.7, 159.2, 161.9 ppm; IR
(KBr): n˜ =3452, 3285, 2965, 2873, 1617, 1502, 1323, 1273, 1160,
1092 cmÀ1; MS (ESI) m/z 480.2 (100) [M+H]+; UPLC purity 100%.
mide (18): Using the synthetic procedure used for compound 12
starting from 11 (87 mg, 0.33 mmol) and 8 (120 mg, 0.33 mmol) to
yield compound 18 as a white powder (130 mg, 72%): Rf =0.25
(CH2Cl2/EtOH 98:2); mp: 77–788C; 1H NMR (400 MHz, [D6]DMSO):
d=0.84 (t, 3H, J=7.2 Hz), 1.51 (sext, 2H, J=7.2 Hz), 2.08 (s, 3H),
2.45 (t, 2H, J=7.2 Hz), 2.76 (d, 1H, J=14.0 Hz), 3.02 (d, 1H, J=
14.0 Hz), 3.45 (d, 1H, J=14.0 Hz), 3.64 (d, 1H, J=14.0 Hz), 4.51 (d,
1H, J=13.2 Hz), 4.57 (d, 1H, J=13.2 Hz), 5.79 (s, 1H), 6.95–7.06 (m,
5H), 7.12 (ddd, 1H, JH–F =J’H–F =9.2 Hz, JH–H =2.4 Hz), 7.22 (d, 2H,
J=7.6 Hz), 7.42 (ddd, 1H, JH–H =8.4 Hz, JH–F =J’H–F =6.8 Hz), 7.61 (d,
2H, J=7.6 Hz), 7.78 (s, 1H), 8.31 (s, 1H), 10.09 ppm (s, 1H);
13C NMR (100 MHz, [D6]DMSO): d=13.7, 24.1, 36.6, 43.9, 56.0, 62.1,
63.2, 75.5, 103.9, 110.8, 120.7 (2C), 126.1, 126.6 (2C), 129.1 (4C),
130.1, 135.4, 138.2, 138.3, 144.7, 145.1, 150.1 ppm, (CF not visible);
IR (KBr): n˜ =3455, 2929, 2860, 1615, 1507, 1460, 1335, 1277, 1159,
1093 cmÀ1; MS (ESI) m/z (%): 556.1 (100) [M+H]+; UPLC purity
99%.
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)-N-(isopropyl)benzenesulfonamide (15):
Using the synthetic procedure used for compound 12 starting
from 11 (130 mg, 0.49 mmol) and 5 (142 mg, 0.49 mmol) to yield
compound 15 as a white powder (mg, 80%): Rf =0.20 (CH2Cl2/
EtOH 98:2); mp: 135–1368C; 1H NMR (400 MHz, [D6]DMSO): d=
0.95 (d, 6H, J=6.4 Hz), 2.13 (s, 3H), 2.79 (d, 1H, J=13.7 Hz), 3.06
(d, 1H, J=13.7 Hz), 3.25 (m, 1H), 3.50 (d, 1H, J=13.4 Hz), 3.69 (d,
1H, J=13.4 Hz), 4.51 (d, 1H, J=14.3 Hz), 4.60 (d, 1H, J=14.3 Hz),
5.82 (s, 1H), 7.00 (ddd, 1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.18
(ddd, 1H, JH–F =J’H–F =9.2 Hz, JH–H =2.4 Hz), 7.29 (d, 2H, J=8.0 Hz),
7.45 (ddd, 1H, JH–H =8.4 Hz, JH–F =J’H–F =6.8 Hz), 7.54 (d, 1H, J=
7.3 Hz), 7.70 (d, 2H, J=8.0 Hz), 7.78 (s, 1H), 8.32 ppm (s, 1H);
13C NMR (100 MHz, [D6]DMSO): d=23.3 (2C), 44.0, 45.4, 56.0, 62.1,
63.2, 75.5, 103.9, 110.8, 126.1, 126.4 (2C), 129.2 (2C), 130.1 (CH),
140.5, 144.0, 145.1, 150.7, 159.2, 161.9 ppm; IR (KBr): n˜ =3440,
3281, 2970, 1616, 1504, 1429, 1316, 1148 cmÀ1; MS (ESI) m/z (%):
480.0 (100) [M+H]+; UPLC purity 95%.
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)-N-(4-isopropylphenyl)benzenesulfon-
amide (19): Using the synthetic procedure used for compound 12
starting from 11 (87 mg, 0.33 mmol) and 9 (120 mg, 0.33 mmol) to
yield compound 19 as a white powder (135 mg, 75%): Rf =0.20
(CH2Cl2/EtOH 98:2); mp: 67–688C; 1H NMR (400 MHz, [D6]DMSO):
d=1.13 (d, 6H, J=6.8 Hz), 2.08 (s, 3H), 2.74–2.82 (m, 2H), 3.03 (d,
1H, J=13.6 Hz), 3.45 (d, 1H, J=14.4 Hz), 3.65 (d, 1H, J=14.4 Hz),
4,51 (d, 1H, J=14.0 Hz), 4.57 (d, 1H, J=14.0 Hz), 5.79 (s, 1H), 6.97
(m, 1H), 7.00 (d, 2H, J=8.0 Hz), 7.10 (m, 3H), 7.23 (d, 2H, J=
7.6 Hz), 7.42 (ddd, 1H, JH–H =8.4 Hz, JH–F =J’H–F =6.8 Hz), 7.63 (d, 2H,
J=7.6 Hz), 7.77 (s, 1H), 8.31 (s, 1H), 10.12 ppm (s, 1H); 13C NMR
(100 MHz, [D6]DMSO): d=24.9 (2C), 32.8, 44.0, 56.0, 62.1, 63.2,
75.5, 103.9, 110.8, 120.7 (2C), 126.2, 126.6 (2C), 127.0 (2C), 129.1
(2C), 130.0, 135.5, 138.4, 144.4, 144.7, 145.1, 150.1, 159.1,
161.8 ppm; IR (KBr): n˜ =3448, 2960, 2864, 1616, 1508, 1461, 1327,
1275, 1160, 1092 cmÀ1; MS (ESI) m/z (%): 556.1 (100) [M+H]+; UPLC
purity 97%.
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)-N-(isobutyl)benzenesulfonamide (16):
Using the synthetic procedure used for compound 12 starting
from 11 (131 mg, 0.49 mmol) and 6 (150 mg, 0.49 mmol) to yield
compound 16 as a colorless oil (195 mg, 81%): Rf =0.25 (CH2Cl2/
EtOH 98:2); 1H NMR (400 MHz, [D6]DMSO): d=0.82 (d, 6H, J=
6.7 Hz), 1.63 (m, 1H), 2.12 (s, 3H), 2.78 (d, 1H, J=13.7 Hz), 3.07 (d,
1H, J=13.7 Hz), 3.35 (m, 2H), 3.49 (d, 1H, J=13.7 Hz), 3.70 (d, 1H,
J=13.7 Hz), 4.52 (d, 1H, J=14.3 Hz), 4.60 (d, 1H, J=14.3 Hz), 5.82
(s, 1H), 7.00 (ddd, 1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.18 (ddd,
1H, JH–F =J’H–F =9.2 Hz, JH–H =2.4 Hz), 7.28 (d, 2H, J=8.5 Hz), 7.45
(ddd, 1H, JH–H =8.4 Hz, JH–F =J’H–F =6.8 Hz), 7.56 (d, 1H, J=6.1 Hz),
7.68 (d, 2H, J=8.5 Hz), 7.79 (s, 1H), 8.32 ppm (s, 1H); 13C NMR
(100 MHz, [D6]DMSO): d=20.1 (2C), 28.2, 44.0, 50.2, 56.0, 62.1,
63.1, 75.5, 103.9, 110.8, 126.2, 126.4 (2C), 129.2 (2C), 130.2, 139.3,
144.1, 145.1, 150.7, 159.2, 161.9 ppm; IR (NaCl): n˜ =3267, 2937,
1600, 1472, 1313, 1138 cmÀ1; MS (ESI) m/z (%): 494.0 (100) [M+H]+;
UPLC purity 97%.
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)-N-(4-nitrophenyl)benzenesulfonamide
(20): Using the synthetic procedure used for compound 12 starting
from 11 (145 mg, 0.54 mmol) and 10 (200 mg, 0.54 mmol) to yield
compound 20 as a yellow powder (125 mg, 33% in two steps):
Rf =0.25 (CH2Cl2/EtOH 98:2); mp: 96–978C; 1H NMR (400 MHz,
[D6]DMSO): d=2.09 (s, 3H), 2.75 (d, 1H, J=13.6 Hz), 3.02 (d, 1H,
J=13.6 Hz), 3.47 (d, 1H, J=14.4 Hz), 3.65 (d, 1H, J=14.4 Hz), 4.48
(d, 1H, J=14.0 Hz), 4.56 (d, 1H, J=14.0 Hz), 5.79 (s, 1H), 6.96 (ddd,
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)-N-(phenyl)benzenesulfonamide (17):
Using the synthetic procedure used for compound 12 starting
from 11 (82 mg, 0.31 mmol) and 7 (100 mg, 0.31 mmol) to yield
compound 17 as a light brown powder (130 mg, 82%): Rf =0.20
(CH2Cl2/EtOH 98:2); mp: 128–1298C; 1H NMR (400 MHz,
[D6]DMSO): d=2.08 (s, 3H), 2.76 (d, 1H, J=13.6 Hz), 3.03 (d, 1H,
J=13.6 Hz), 3.45 (d, 1H, J=14.4 Hz), 3.64 (d, 1H, J=14.4 Hz), 4.51
(d, 1H, J=14.0 Hz), 4.57 (d, 1H, J=14.0 Hz), 5.79 (s, 1H), 6.97 (ddd,
1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.04 (t, 1H, J=7.2 Hz), 7.09–
1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.11 (ddd, 1H, JH–F =J’H–F
9.2 Hz, JH–H =2.4 Hz), 7.29–7.33 (m, 4H), 7.41 (ddd, 1H, JH–H =8.4 Hz,
H–F =J’H–F =6.8 Hz), 7.76–7.78 (m, 3H), 8.16 (d, 2H, J=8.8 Hz), 8.29
=
J
(s, 1H), 11.26 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=44.0,
56.0, 62.0, 63.2, 75.5, 103.7, 111.0, 118.1 (2C), 125.6 (2C), 126.1,
126.7 (2C,), 129.5 (2C), 130.1, 137.6, 142.7, 144.5, 145.1, 145.6,
150.6 ppm, (CF not visible); IR (KBr): n˜ =3444, 1599, 1500, 1342,
1276, 1161, 1090 cmÀ1; MS (ESI) m/z (%): 559.0 (100) [M+H]+; UPLC
purity 99%.
7.16 (m, 3H), 7.23–7.26 (m, 4H), 7.42 (ddd, 1H, JH–H =8.4 Hz, JH–F
=
J’H–F =6.8 Hz), 7.64 (d, 2H, J=8.0 Hz), 7.78 (s, 1H), 8.31 (s, 1H),
10.24 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=43.9, 56.0,
62.1, 63.3, 75.5, 103.8, 110.7, 120.2 (2C), 124.2, 126.2, 126.6 (2C),
129.1 (2C), 129.3 (2C), 130.1, 137.9, 138.1, 144.8, 145.1, 150.6 ppm,
(CF not visible); IR (KBr): n˜ =3465, 1613, 1498, 1339, 1271, 1159,
1092 cmÀ1; MS (ESI) m/z (%): 514.1 (100) [M+H]+; UPLC purity
95%.
2-(2,4-Difluorophenyl)-1-[methyl(4-nitrobenzyl)amino]-3-(1H-
1,2,4-triazol-1-yl)propan-2-ol (21): Using the synthetic procedure
used for compound 12 starting from 11 (1.24 g, 4.63 mmol) and 4-
nitrobenzylbromide (1.00 g, 4.63 mmol) to yield compound 21 as
an orange oil (1.44 g, 77%): Rf =0.25 (CH2Cl2/EtOH 98:2); 1H NMR
(400 MHz, [D6]DMSO): d=2.13 (s, 3H), 2.79 (d, 1H, J=13.7 Hz),
3.10 (d, 1H, J=13.7 Hz), 3.56 (d, 1H, J=14.3 Hz), 3.77 (d, 1H, J=
14.3 Hz), 4.53 (d, 1H, J=14.0 Hz), 4.61 (d, 1H, J=14.0 Hz), 5.85 (s,
1H), 7.00 (ddd, 1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.19 (ddd, 1H,
4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)pro-
pyl]methylamino}methyl)-N-(4-propylphenyl)benzenesulfona-
822
www.chemmedchem.org
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
ChemMedChem 2011, 6, 816 – 825

Antifungal Agents
J
H–F =J’H–F =9.2 Hz, JH–H =2.4 Hz), 7.38 (d, 2H, J=8.5 Hz), 7.46 (ddd,
1H, JH–H =8.4 Hz, JH–F =J’H–F =6.8 Hz), 7.56 (d, 2H, J=7.6 Hz), 7.63
(s, 1H), 7.75–7.77 (m, 3H), 8.30 (s, 1H), 10.19 ppm (s, 1H); 13C NMR
(100 MHz, [D6]DMSO): d=43.8, 56.0, 62.2, 63.0, 75.2, 103.8, 110.7,
120.2 (2C), 126.3, 126.8 (2C), 129.4 (2C), 129.5 (2C), 130.0, 133.0,
135.0, 136.5, 139.7, 145.0, 150.6 ppm, (CF not visible); IR (KBr): n˜ =
3450, 1619, 1506, 1331, 1158 cmÀ1; MS (ESI) m/z (%): 514.0 (100)
[M+H]+; UPLC purity 95%.
1H, JH–H =8.4 Hz, JH–F =J’H–F =6.8 Hz), 7.79 (s, 1H), 8.13 (d, 2H, J=
8.5 Hz), 8.32 ppm (s, 1H); IR (NaCl): n˜ =3381, 2948, 1609, 1514,
1421, 1347, 1272, 1102 cmÀ1. MS (ESI) m/z (%): 404.0 (100) [M+H]+;
UPLC purity 100%.
2-(2,4-Difluorophenyl)-1-[methyl(4-aminobenzyl)amino]-3-(1H-
1,2,4-triazol-1-yl)propan-2-ol (22): Active charcoal-supported Pd
(5%, 140 mg) was added to a stirred solution of 21 (1.4 g,
3.47 mmol) in EtOH (100 mL). The solution was stirred at RT for 2 h
under H2 atmosphere (5 bar). The mixture was filtered through a
pad of Celite, and the filtrate was concentrated. The residue was
purified using silica gel column chromatography (CH2Cl2 and
CH2Cl2/EtOH 98:2) to give compound 22 as a light brown powder
N-[4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)-
propyl]methylamino}methyl)phenyl]-4-isopropylbenzenesulfon-
amide (25): Using the synthetic procedure used for compound 24
starting from with 22 (100 mg, 0.27 mmol) and 4-isopropylbenze-
nesulfonyle chloride (48 mL, 0.27 mmol) to yield compound 25 as a
light brown powder (100 mg, 67%): Rf =0.50 (CH2Cl2/EtOH 95:5);
mp: 92–938C; 1H NMR (400 MHz, [D6]DMSO): d=1.19 (d, 6H, J=
6.8 Hz), 2.04 (s, 3H), 2.73 (d, 1H, J=13.6 Hz), 2.93–2.98 (m, 2H),
3.31 (d, 1H, J=13.2 Hz), 3.45 (d, 1H, J=13.2 Hz), 4.48 (d, 1H, J=
14.4 Hz), 4.54 (d, 1H, J=14.4 Hz), 5.72 (s, 1H), 6.94–7.01 (m, 5H),
7.11 (ddd, 1H, JH–F =J’H–F =9.2 Hz, JH–H =2.4 Hz), 7.38–7.44 (m, 3H),
7.69 (d, 2H, J=8.4 Hz), 7.76 (s, 1H), 8.30 (s, 1H), 10.19 ppm (s, 1H);
13C NMR (100 MHz, [D6]DMSO): d=23.5 (2C), 33.5, 43.8, 56.0, 62.2,
63.2, 75.2, 103.8, 110.7, 119.8 (2C), 126.3, 126.9 (2C), 127.3 (2C),
129.6 (2C), 130.0, 134.8, 136.7, 137.4, 145.0, 150.6, 153.7 ppm, (CF
not visible); IR (KBr): n˜ =3450, 2958, 1618, 1508, 1328, 1162,
1087 cmÀ1; MS (ESI) m/z (%): 556.2 (100) [M+H]+; UPLC purity
98%.
1
(940 mg, 72%): Rf =0.40 (CH2Cl2/EtOH 95:5); mp: 86–878C; H NMR
(400 MHz, [D6]DMSO): d=2.07 (s, 3H), 2.78 (d, 1H, J=14.0 Hz),
2.93 (d, 1H, J=14.0 Hz), 3.26 (d, 1H, J=14.0 Hz), 3.34 (d, 1H, J=
14.0 Hz), 4.48 (d, 1H, J=14.3 Hz), 4.56 (d, 1H, J=14.3 Hz), 4.95
(s, 2H), 5.70 (s, 1H), 6.47 (d, 2H, J=8.2 Hz), 6.77 (d, 2H, J=8.2 Hz),
6.98 (ddd, 1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.17 (ddd, 1H,
J
H–F =J’H–F =9.2 Hz, JH–H =2.4 Hz), 7.43 (ddd, 1H, JH–H =8.4 Hz, JH–F =
J’H–F =6.8 Hz), 7.75 (s, 1H), 8.31 ppm (s, 1H); IR (KBr): n˜ =3308,
3211, 1617, 1507, 1280, 1129 cmÀ1; MS (ESI) m/z (%): 105.7 (100),
374.1 (12) [M+H]+.
N-[4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)-
propyl]methylamino}methyl)phenyl]propane-2-sulfonamide (23):
Isopropylsulfonyl chloride (49 mL, 0.44 mmol) was added to a
stirred solution of 22 (150 mg, 0.40 mmol) in pyridine (2 mL). The
solution was stirred at RT for 2 h. The solvent was removed under
reduced pressure and the residue was partitioned between CH2Cl2
and H2O. Organic layers were dried over anhydrous Na2SO4 and
concentrated in vacuo. The residue was purified using silica gel
column chromatography (CH2Cl2 and CH2Cl2/EtOH 99:1) to give
compound 23 as a white powder (90 mg, 47%): Rf =0.10 (CH2Cl2/
N-[4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)-
propyl]methylamino}methyl)phenyl]-4-nitrobenzenesulfonamide
(26): Using the synthetic procedure used for compound 24 starting
from 22 (150 mg, 0.40 mmol) and 4-nitrobenzenesulfonyle chloride
(89 mg, 0.40 mmol) to yield compound 26 as a light yellow
powder (65 mg, 29%): Rf =0.10 (CH2Cl2/EtOH 98:2); mp: 159–
1
1608C; H NMR (400 MHz, [D6]DMSO): d=2.07 (s, 3H), 2.72 (d, 1H,
J=14.0 Hz), 2.93 (d, 1H, J=14.0 Hz), 3.33 (d, 1H, J=13.2 Hz), 3.45
(d, 1H, J=13.2 Hz), 4.42 (d, 1H, J=14.0 Hz), 4.54 (d, 1H, J=
1
EtOH 98:2); mp: 60–618C; H NMR (400 MHz, [D6]DMSO): d=1.24
14.0 Hz), 5.73 (s, 1H), 6.94–7.00 (m, 5H), 7.12 (ddd, 1H, JH–F =J’H–F
9.2 Hz, H–H =2.4 Hz), 7.40 (ddd, 1H, H–H =8.4 Hz, H–F =J’H–F
=
=
(d, 6H, J=6.8 Hz), 2.10 (s, 3H), 2.77 (d, 1H, J=13.6 Hz), 2.98 (d, 1H,
J=13.6 Hz), 3.20 (m, 1H), 3.40 (d, 1H, J=13.2 Hz), 3.50 (d, 1H, J=
13.2 Hz), 4.47 (d, 1H, J=14.4 Hz), 4.56 (d, 1H, J=14.4 Hz), 5.74 (s,
1H), 7,00 (ddd, 1H, JH–F =JH–H =8.4 Hz, JH–H =2.4 Hz), 7.04 (d, 2H,
J=8.8 Hz), 7.14 (d, 2H, J=8.8 Hz), 7.17 (ddd, 1H, JH–F =J’H–F
9.2 Hz, H–H =2.4 Hz), 7.44 (ddd, 1H, H–H =8.4 Hz, H–F =J’H–F
J
J
J
6.8 Hz), 7.75 (s, 1H), 7.99 (d, 2H, J=8.8 Hz), 8.30 (s, 1H), 8.39 (d,
2H, J=8.8 Hz), 10.54 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO):
d=43.9, 55.6, 61.8, 62.8, 75.3, 103.5, 110.5, 120.8 (2C), 124.8 (2C),
126.4, 128.4 (2C), 129.8 (2C), 130.1, 135.7, 135.8, 145.0, 145.1, 150.6,
150.1 ppm, (CF not visible); IR (KBr): n˜ =3450, 1623, 1528, 1350,
1166 cmÀ1; MS (ESI) m/z (%): 559.2 (100) [M+H]+; UPLC purity 96%.
=
=
J
J
J
6.8 Hz), 7.77 (s, 1H), 8.31 (s, 1H), 9.70 ppm (s, 1H); 13C NMR
(100 MHz, [D6]DMSO): d=16.3 (2C), 43.9, 51.3, 56.0, 62.2, 63.0,
75.1, 103.7, 110.9, 119.4 (2C), 126.4, 129.8 (2C), 130.1, 134.4, 137.4,
145.0, 150.6 ppm, (CF not visible); IR (KBr): n˜ =3445, 1620, 1508,
1326, 1140 cmÀ1; MS (ESI) m/z (%): 480.0 (100) [M+H]+; UPLC
purity 95%.
Biological assays
Antifungal activity: All these compounds were screened for their
antifungal activity against C. albicans (CA98001, DSY735, CAAL-74),
C. krusei (CK506, CK8), C. glabrata (CG468), C. parapsilosis (CAPA1,
CAPA2), and A. fumigatus (AF98003) strains. DSY735 was a gift
from Pr. D. Sanglard (Institute of Microbiology, University of Lau-
sanne, Switzerland). All other strains were issued from our collec-
tion. Inhibition growth was measured as previously described.[46]
Fluconazole, voriconazole, and itraconazole were used as positive
controls. MICs were defined as the concentrations that inhibit
growth of Candida spp. or A. fumigatus by 50% or 80% respective-
ly as recommended by the Clinical and Laboratory Standards Insti-
tute document M27A3 and M38A2 guidelines.[47] MIC values are
N-[4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)-
propyl]methylamino}methyl)phenyl]benzenesulfonamide
(24):
Indium powder (5 mg, 0.04 mmol) was added to a stirred solution
of 22 (150 mg, 0.40 mmol) in CH3CN (3 mL) under argon followed
by the addition of benzenesulfonyle chloride (52 mL, 0.40 mmol).
The solution was stirred at RT overnight. The solvent was removed
under reduced pressure and the residue was partitioned between
CH2Cl2 and H2O. Organic layers were dried over anhydrous Na2SO4
and concentrated in vacuo. The residue was purified using silica
gel column chromatography (CH2Cl2 and CH2Cl2/EtOH 99:1) to
yield compound 24 as a white powder (140 mg, 68%): Rf =0.15
(CH2Cl2/EtOH 98:2); mp: 125–1268C; 1H NMR (400 MHz,
[D6]DMSO): d=2.04 (s, 3H), 2.73 (d, 1H, J=13.6 Hz), 2.95 (d, 1H,
J=13.6 Hz), 3.31 (d, 1H, J=13.2 Hz), 3.44 (d, 1H, J=13.2 Hz), 4.47
(d, 1H, J=14.4 Hz), 4.54 (d, 1H, J=14.4 Hz), 5.72 (s, 1H), 6.94–7.00
(m, 5H), 7.13 (ddd, 1H, JH–F =J’H–F =9.2 Hz, JH–H =2.4 Hz), 7.41 (ddd,
expressed in mgmLÀ1
.
Sterol extraction and analysis: To study sterol synthesis, C. albicans
CA98001 cells were incubated in 50 mL Sabouraud broth medium
(Sigma–Aldrich) for 18 h at 358C with stirring. Compound 17 was
introduced into the culture medium before incubation. Cells were
ChemMedChem 2011, 6, 816 – 825
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemmedchem.org
823

C. Logꢀ et al.
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Published online on March 2, 2011
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ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemmedchem.org
825