Design, Synthesis, and Biological Evaluation of 1-[(Biarylmethyl)methylamino]-2-(2,4-difluorophenyl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ols as Potent Antifungal Agents: New Insights into Structure-Activity Relationships

Article abstract of DOI:10.1002/cmdc.201100262

We recently reported the design and synthesis of azole antifungal agents with a focus on modifications to the side chain appended to the propanol group. Herein we have identified a series of new 1-[(biarylmethyl)methylamino] derivatives with broad-spectrum antifungal activities against the most prevalent human pathogenic fungi (Candida spp. and Aspergillus fumigatus). Compounds containing a flexible benzylamine moiety were clearly shown to yield the best antifungal activities, without the need for a hydrogen-bond acceptor substituent directly attached to the para position. We were also able to determine that selected compounds are able to overcome gene overexpression and point mutations that lead to reduced susceptibility or resistance against current treatments, such as fluconazole. As the minor differences observed with small structural modifications cannot be explain with only a three-dimensional model of CYP51, adequate physicochemical parameters must be evaluated in terms of antifungal potency, bioavailability, and toxicity. Therefore, structure-activity relationship studies such as these reveal new insights for the development of future antifungal therapies. Attacking the fungus among us: 1-[(Biarylmethyl)methylamino] derivatives with broad-spectrum antifungal activities against the most prevalent human pathogenic fungi (Candida spp. and Aspergillus fumigatus) are described, revealing further information about their structure-activity relationships.

Full text of DOI:10.1002/cmdc.201100262

MED
DOI: 10.1002/cmdc.201100262
Design, Synthesis, and Biological Evaluation of
1-[(Biarylmethyl)methylamino]-2-(2,4-difluorophenyl)-3-
(1H-1,2,4-triazol-1-yl)propan-2-ols as Potent Antifungal
Agents: New Insights into Structure–Activity Relationships
Rꢀmi Guillon,[a] Fabrice Pagniez,[b] Charlotte Rambaud,[a] Carine Picot,[b] Muriel Duflos,[a]
Cꢀdric Logꢀ,*[a] and Patrice Le Pape[b]
We recently reported the design and synthesis of azole anti-
fungal agents with a focus on modifications to the side chain
appended to the propanol group. Herein we have identified a
series of new 1-[(biarylmethyl)methylamino] derivatives with
broad-spectrum antifungal activities against the most preva-
lent human pathogenic fungi (Candida spp. and Aspergillus fu-
to determine that selected compounds are able to overcome
gene overexpression and point mutations that lead to reduced
susceptibility or resistance against current treatments, such as
fluconazole. As the minor differences observed with small
structural modifications cannot be explain with only a three-di-
mensional model of CYP51, adequate physicochemical parame-
ters must be evaluated in terms of antifungal potency, bioavail-
ability, and toxicity. Therefore, structure–activity relationship
studies such as these reveal new insights for the development
of future antifungal therapies.
migatus). Compounds containing
a flexible benzylamine
moiety were clearly shown to yield the best antifungal activi-
ties, without the need for a hydrogen-bond acceptor substitu-
ent directly attached to the para position. We were also able
Introduction
Invasive fungal infections (IFIs) are a growing threat to human
health. These infections have become increasingly common
and predominantly occur in the context of aggressive immu-
nosuppressive therapies, such as anticancer chemotherapy,
organ transplants, or acquired immune deficiency syndrome
(AIDS). The most prominent fungal pathogens affecting
humans belong to Candida and Aspergillus species, and overall
mortality for these infections remains at 40–80%.[1] Currently,
the main classes of antifungal drugs for treatment of IFIs are
polyenes, azoles, and echinocandins, which target components
of either the fungal membrane or cell wall.[2] However, despite
a larger therapeutic armamentarium, problems such as intrinsic
or acquired antifungal resistance remain, which require re-
searchers to develop new antifungal drugs with expanded ef-
fectiveness.[3]
Although CYP51 enzymes from C. albicans and A. fumigatus
have recently been expressed in Escherichia coli and purified
from the membrane fraction to investigate azole binding prop-
erties,[13,14] and numerous crystal structures of CYP51 from
human and Trypanosomatidae in complex with azole antifungal
inhibitors have been elucidated,[15,16] no experimental structural
information from pathogenic fungi is yet available. This type of
information is of fundamental importance to study azole bind-
ing interactions and to improve the characteristics of these
compounds in a rational design approach. To date, only three-
dimensional models of CYP51 from C. albicans, A. fumigatus,
and Cryptococcus neoformans were constructed by homology
modeling,[17–20] and the binding modes of antifungal agents
were investigated. Of note, Sheng et al. constructed a pharma-
cophore model on the basis of the C. albicans enzyme and
highlighted the importance of both Tyr118 and Ser378 for
p–p stacking and hydrogen-bonding interactions in the stabili-
Resistance to triazoles is the most serious concern, as these
are commonly the antifungal agents used to treat Candida
species. These drugs (fluconazole, voriconazole, itraconazole,
and posaconazole) mainly target P450-dependent sterol 14a-
demethylase (CYP51, Erg11), encoded by the ERG11 gene, a
key enzyme in fungal ergosterol biosynthesis.[4] Azole resist-
ance in C. albicans has been associated with genetic alterations
in the ERG11 gene, leading to amino acid substitutions in the
target enzyme CYP51 that reduce drug binding.[5–7] Resistance
can also be the result of other mechanisms, such as overex-
pression of the CDR1/2 and MDR1 genes that encode drugs-
efflux pumps,[8,9] development of pathways that bypass target
enzymes,[10,11] and ERG11 gene overexpression.[12]
[a] Dr. R. Guillon, C. Rambaud, 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, Nantes 44035 Cedex 1 (France)
E-mail: cedric.loge@univ-nantes.fr
[b] Dr. F. Pagniez, C. Picot, 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, Nantes 44035 Cedex 1 (France)
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Antifungal Agents
zation of inhibitors.[21–24] Furthermore, Xiao et al. suggested a
model for the binding of azoles with extended side chains in
CYP51 from C. albicans and A. fumigatus. Specifically, itracon-
azole and posaconazole appear to be less affected by substitu-
tions near the heme site than either fluconazole or voricon-
azole, which could be explained by tighter affinity and/or com-
pensatory adjustments within the active site.[17]
Taking into account that series with hydrogen-bond accept-
ors in the para position of a flexible benzyl group lead to
broad-spectrum compounds, we decided to pursue new ben-
zylamine derivatives with extended side chains, including addi-
tional aromatic rings (benzene, pyridine, or pyrazole) in the
meta or para positions (Figure 1). With a diverse selection of
“probe” substituents (methoxy, cyano, nitro, trifloromethyl,
morpholine, N-methylpiperazine, and sulfonamides), this series
has also provided new insights into structure–activity relation-
ships (SAR).
In recent years, most of the work in azole optimization has
been focused on modification of the side chain attached to
the propanol group,[20,21,25,26] and we also reported the synthe-
sis of multiple series of antifungal agents with varying degrees
of antifungal activity against C. albicans and A. fumigatus
strains.[27–31] Among our molecules, the recent series of 4-({2- Results and Discussion
(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl]-
Chemistry
methylamino}methyl)benzenesulfonamides (I, Figure 1) yielded
Synthesis of target compounds 4–14 began from derivative
1,[29] which was combined with 4-iodobenzylbromide or 3-io-
dobenzylbromide in presence of Hunig’s base to form inter-
mediates 2 and 3 (Scheme 1). Suzuki cross-coupling with vari-
ous para-substituted (hetero)arylboronic acids (or esters) af-
forded molecules 4–14. Phenylboronic acid and 4-methoxy-
phenylboronic acid were coupled using standard reaction con-
ditions, whereas introduction of aryls with electron-
withdrawing substituents was performed using microwave irra-
diation.[32]
Figure 1. Lead structure I (left) reported in Reference [31] and general struc-
tures of synthesized compounds (right). For the structures of the aromatic
moiety (Ar), see Table 1.
The synthetic route to target compounds 15–17 and 19–21
is outlined in Scheme 2. 2-Bromopyridine-5-boronic acid was
prepared as described in the literature[33] and utilized in Suzuki
cross-coupling with derivative 2 to afford derivative 15 in 60%
yield.[34] Molecules 16 and 17 were obtained from 15 by opti-
mization of Buchwald–Hartwig amination conditions with N-
methylpiperazine or morpholine using Pd2(dba)3, tBuONa, and
2,2’-bis(diphenylphosphino)-1,1’-binaphthyl (BINAP) in toluene.
Under the same conditions, benzophenone imine was coupled
with 15 to afford intermediate 18, which was deprotected with
the most active compounds against Candida spp. and A. fumi-
gatus strains, consistent with the pharmacophore model.[31]
These compounds also demonstrated the capacity to over-
come upregulation of CDR and ERG11 genes and were able to
maintain antifungal activity despite a recognized and critical
CYP51 substitution in C. albicans isolates.
Scheme 1. Reagents and conditions: a) 4-iodobenzylbromide or 3-iodobenzylbromide, N,N-diisopropylethylamine, CH3CN, RT, 24 h, 54–92%; b) benzeneboronic
acid or 4-methoxybenzeneboronic acid, Pd(PPh3)4, 2m aq Na2CO3, EtOH, toluene, reflux, 3 h, 56–75%; c) boronic acid or boronic acid pinacol ester, Pd(PPh3)4,
2m aq Na2CO3, CH3CN, microwave 100 W, 1208C, 10 min, 41–70%.
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Scheme 2. Reagents and conditions: a) 2-bromopyridine-5-boronic acid, Pd(PPh3)4, 1m aq Na2CO3, DMF, 908C, 2 h, 60%; b) amines or benzophenone imine,
Pd2(dba)3, BINAP, tBuONa, toluene, 1008C, 2 h, 70–82%; c) hydroxylamine hydrochloride, AcONa, MeOH, RT, 2 h, 82%; d) R2SO2Cl, pyridine, 558C, 2 h, 73–76%.
hydroxylamine hydrochloride to give amine 19 in good yield.
Finally, reaction of 19 with methane- or benzenesulfonylchlor-
ides in pyridine led to compounds 20 and 21.
unsubstituted biphenyl molecule 4 (MIC <0.001 ngmLÀ1) was
as active toward the C. albicans strain as compounds 5–8,
which have additional electron-donating or electron-withdraw-
ing groups, suggesting that these substituents have only
minor influence. Additionally, the biological results obtained
for compounds 9–11, which contain pyridine or pyrazole moi-
eties, were comparable to those for derivative 4, showing that
the presence of such azaheterocycles is not necessary for
potent antifungal activity against C. albicans.
Antifungal activity
The in vitro antifungal activities of all compounds were evalu-
ated against human pathogenic fungi (C. albicans, C. krusei,
C. glabrata, C. parapsilosis, and A. fumigatus) and are summar-
ized in Table 1. Minimum inhibitory concentration (MIC)values
of fluconazole, voriconazole, and itraconazole are shown as ref-
erence compounds.
Sterol analysis
Regardless of the aromatic rings linked the benzylamine
group or the size and nature of various substituents, all of the
synthesized compounds displayed a high level of activity
toward the C. albicans CA98001 strain, with MIC values ranging
from 1.0 to 20 ngmLÀ1 and comparable to that of voriconazole.
Significant broad-spectrum antifungal activity was also ob-
served for the majority of these molecules against C. krusei
CK8, C. glabrata, and C. parapsilosis strains, confirming their po-
tential against Candida strains with low susceptibility or intrin-
sic resistance to fluconazole. Compounds 6 and 7, with either
a 4-cyano or a 4-nitrophenyl substituent, exhibited the most
potent antifungal activity against the C. krusei CK506 strain
(MIC values of 1.32 and 1.60 mgmLÀ1, respectively). A majority
of the compounds (4–11 and 15–19) exhibited promising bio-
logical results toward the A. fumigatus AF98003 strain, with
MIC values ranging from 2.2 to 3.6 mgmLÀ1, only five- to nine-
fold higher than that of itraconazole.
The mechanism of action for this series of azoles was investi-
gated by studying inhibition of C. albicans CA98001 ergosterol
biosynthesis following treatment with 6, the most active com-
pound. As shown in Table 2, 60% inhibition of ergosterol bio-
synthesis was obtained with a concentration of 4.59 ngmLÀ1
,
near the MIC value (1.0 ngmLÀ1), and lanosterol accumulation
was observed. At a higher concentration (22.98 ngmLÀ1), this
effect was maximal with virtually undetectable production of
ergosterol. Depletion of ergosterol and significant accumula-
tion of 14-methyl-3,6-diol (a well-known toxic metabolite ob-
tained from 14-methylfecosterol and catalyzed by the D5,6-de-
saturase encoded by the ERG3 gene)[35] are typically the results
of growth inhibition following azole treatment and confirm
inhibition of the CYP51 enzyme.
Biological evaluation against C. albicans isolates with
reduced fluconazole susceptibility
Compounds 12–14 are of particular interest, as compared to
their immediate analogues 4, 6, and 9. Overall, the introduc-
tion of aromatic substituents at the para versus the meta posi-
tion of the benzylamine group yielded less active compounds,
especially against A. fumigatus, C. parapsilosis, and C. glabrata
strains. The biological activities against the C. albicans strain
were less sensitive to these steric changes. Interestingly, the
The biological effects of these compounds were also investi-
gated against two C. albicans strains with known mechanisms
of resistance (DSY735 and CAAL-74).[31] Compounds 6 and 7
were selected for their broad-spectrum antifungal activity, with
MIC values presented in Table 3. Both compounds exhibited
high antifungal activities against these strains, with MIC values
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Table 1. In vitro antifungal activities of compounds 4–14, 15–17, and 19–21 against Candida spp. and A. fumigatus strains.
[a]
Compd
Ar
MIC values [mgmLÀ1
]
CA98001
<0.001
0.001Æ0.001 >40
CK506
CK8
CG468
CAPA1
CAPA2
AF98003
4
5
6
7
8
>40
1.55Æ0.06
0.97Æ0.25
0.154Æ0.001
0.156Æ0.002
0.152Æ0.035
0.167Æ0.014
0.092Æ0.009
0.149Æ0.014
1.180Æ0.151
0.143Æ0.008
0.110Æ0.032
0.151Æ0.030
0.134Æ0.043
1.11Æ0.27
2.66Æ0.13
2.36Æ0.05
2.76Æ0.09
3.16Æ0.05
3.07Æ0.15
0.001Æ0.001
0.002Æ0.001
1.32Æ0.02 0.193Æ0.005 0.133Æ0.009
1.60Æ0.10 0.88Æ0.09
2.40Æ0.50 1.54Æ0.08
0.158Æ0.005
1.52Æ0.09
<0.001
9
0.013Æ0.003
0.001Æ0.001
12.8Æ0.7
13.8Æ0.2
1.9Æ0.1
1.2Æ0.1
0.113Æ0.004
0.122Æ0.004
0.875Æ0.135
0.191Æ0.037
0.196Æ0.078
0.218Æ0.073
2.96Æ0.04
2.22Æ0.04
10
11
0.008Æ0.001
29.4Æ5.0
8.7Æ1.3
1.15Æ0.13
1.263Æ0.175
0.740Æ0.019
2.89Æ0.04
12
13
0.012Æ0.001
0.002Æ0.001
35.8Æ0.1
30.3Æ2.4
3.6Æ1.2
0.786Æ0.434
1.19Æ0.14
1.230Æ0.035
0.873Æ0.184
1.53Æ0.15
1.51Æ0.11
24.7Æ0.9
20.2Æ1.4
0.16Æ0.01
14
15
16
17
19
20
21
0.012Æ0.002 >40
0.002Æ0.001 >50
14.3Æ0.2
1.2Æ0.1
1.54Æ0.12
2.0Æ0.1
1.4Æ0.1
27.9Æ4.1
5.1Æ1.6
1.42Æ0.01
0.139Æ0.001
1.48Æ0.04
11.3Æ1.5
9.42Æ1.34
8.53Æ1.17
>40
0.195Æ0.001
0.080Æ0.021
0.118Æ0.031
0.149Æ0.011
0.078Æ0.026
0.203Æ0.090
2.71Æ0.25
2.98Æ0.05
3.62Æ0.11
2.92Æ0.05
2.93Æ0.09
29.6Æ1.1
0.020Æ0.053
<0.001
15.8Æ0.8
>50
<0.05
0.002Æ0.001
15.3Æ0.4
0.401Æ0.121
12.7Æ2.3
0.369Æ0.203
0.010Æ0.002 >50
1.22Æ0.20
1.14Æ0.12
0.002Æ0.001 >50
0.036Æ0.021 >30
1.91Æ0.14
1.35Æ0.13
>60
Fluconazole
Voriconazole
Itraconazole
–
–
–
12.9Æ0.9
0.24Æ0.07
–
7.7Æ0.1
0.45Æ0.04
–
>30
>30
–
0.005Æ0.001
1.3Æ0.3
0.95Æ0.13
0.36Æ0.01
0.15Æ0.01
0.42Æ0.04
–
–
–
–
[a] Values represent the meanÆSD of experiments performed in triplicate; C. albicans (CA98001), C. krusei (CK506, CK8), C. glabrata (CG468), C. parapsilosis
(CAPA1, CAPA2), and A. fumigatus (AF98003).
Molecular modeling
ranging from 22–91 ngmLÀ1, implying that they are able to
overcome overexpression of CDR and ERG11 genes or specific
point substitutions in the CYP51 enzyme which result in flu-
conazole resistance, as observed in the previously described
benzenesulfonamide series (I, Figure 1).[31]
In an attempt to understand the observed SAR, we first per-
formed docking of compounds 6 (MIC=1.0 ngmLÀ1) and 13
(MIC=2.0 ngmLÀ1) with our homology model of CYP51 from
C. albicans (Figure 2a).[30,36] Despite different side chain confor-
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Table 2. Effect of compound
6 on sterol composition of C. albicans
CA98001.
Sterols[a]
Control
6 [ngmLÀ1
]
4.59
22.98
Lanosterol
Eburicol
Zymosterol
Episterol
14-Methylfecosterol
14-Methylepisterol
14-Methyl-3,6-diol
Ergosterol
6.0
3.4
2.0
3.7
–
–
–
84.7
27.6
21.8
–
14.5
7.9
–
–
–
2.4
6.7
8.8
32.7
7.1
7.2
62.7
0.7
[a] Sterols of interest were identified by mass spectrometry. The area
under the curve (AUC) of each peak was used to calculate a ratio: [sterol
AUC/sum of sterols AUC]ꢃ100.
Table 3. In vitro antifungal activities of compounds 6 and 7 against C. al-
bicans strains with reduced fluconazole susceptibility.
[a]
Compd
Ar
MIC [mgmLÀ1
]
DSY735
CAAL-74
6
0.073Æ0.018
0.022Æ0.004
0.091Æ0.023
7
0.086Æ0.005
Fluconazole
–
12.55
>30
[a] Values represent the meanÆSD of experiments performed in tripli-
cate.
Figure 2. a) Predicted binding of compounds (S)-6 (magenta) and (S)-13
mations, both compounds adopt a similar binding interaction
with the enzyme. The benzonitrile groups are in close proximi-
ty to residues His377, Tyr64, and Phe380 but not to Ser378 (d
>5 ꢂ), the amino acid conserved across the fungal CYP51
enzyme and proposed in the pharmacophore model to form a
key hydrogen bonding interaction with inhibitors.[21]
(orange) in the proposed active site pocket (channel 2) of CYP51 from C. al-
bicans. Hip377 is the protonated form of histidine residue. b) Predicted
binding of compounds (S)-4, (S)-10, and (S)-19 in the proposed active site
pocket (channel 2) of CYP51 from C. albicans. Hydrogen bonds are indicated
as yellow dotted lines.
The docking of representative molecules, such as the non-
substituted biphenyl compound 4 or compounds 10 and 19,
which possess a terminal pyridin-4-yl or a 6-aminopyridin-3-yl
group, respectively, should confirm that the binding of those
azoles to the CYP51–C. albicans enzyme occurs mostly through
hydrophobic interactions (Figure 2b). In fact, even if an addi-
tional hydrogen-bonding interaction seems possible between
the imidazole side chain of His377 and both nitrogen atoms in
the 3- or 4-position of the pyridine ring, the fact that similar
biological results are obtained for compound 4 (which does
not have an electron-withdrawing substituent) clearly demon-
strates the critical importance of key residue Tyr118 and the
class-specific residue Phe380 in p–p stacking interactions with
the bi(hetero)aryl groups of these antifungal agents. Interac-
tions such as these contribute to improved stabilization of the
inhibitors within the active site.
group, this manuscript clearly confirms that compounds con-
taining a flexible benzylamine moiety yielded the best results
against Candida spp. and A. fumigatus strains, without the
need for a hydrogen-bond acceptor substituent directly at-
tached to the para position. Selected biaryl compounds 6 and
7, with a 4-cyano or a 4-nitro substituent, are also able to over-
come overexpression of CDR and ERG11 genes in a C. albicans
strain with reduced susceptibility to fluconazole and can main-
tain activity despite a known CYP51 point mutation from a flu-
conazole-resistant C. albicans strain. However, close attention
must be paid to observed SAR based on MIC data in the ab-
sence of biological results regarding CYP51. Although all of the
synthesized compounds exhibited excellent antifungal activity
against C. albicans, minor differences observed with small
structural modifications cannot be explained with a three-
dimensional model of CYP51 alone. Adequate physicochemical
parameters must be considered in terms of antifungal potency,
bioavailability, and toxicity. In particular, the relatively high lip-
ophilicity observed for these compounds (6, clogP=3.77; 7,
clogP=4.08; octanol–water partition coefficient calculated
Conclusions
Among the several azoles synthesized in our laboratory, with a
focus on modification of the side chain linked to the propanol
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Antifungal Agents
0.01 mmol) was added under argon to a stirred solution of 2
(200 mg, 0.41 mmol) in toluene (2 mL), and the solution was stirred
at RT for 20 min. Benzeneboronic acid (60 mg, 0.50 mmol) in EtOH
(0.1 mL) was added, followed by the addition of 2m aq Na2CO3
(0.248 mL, 0.50 mmol). The resulting mixture was stirred at reflux
for 3 h. The solvent was removed in vacuo, and the residue was
partitioned between CH2Cl2 and H2O. The organic layers were dried
over anhyd Na2SO4 and concentrated in vacuo. The residue was
purified using silica gel column chromatography (CH2Cl2 and
CH2Cl2/EtOH, 99:1) to yield compound 4 as a white powder
(120 mg, 67%): Rf =0.10 (CH2Cl2/EtOH, 98:2); mp: 117–1188C;
1H NMR (400 MHz, [D6]DMSO): d=2.13 (s, 3H), 2.82 (d, 1H, J=
13.2 Hz), 3.08 (d, 1H, J=13.2 Hz), 3.48 (d, 1H, J=13.6 Hz), 3.64 (d,
1H, J=13.6 Hz), 4.55 (d, 1H, J=14.0 Hz), 4.61 (d, 1H, J=14.0 Hz),
5.80 (s, 1H), 7.01 (ddd, 1H, JHÀF =JHÀH =8.4 Hz, JHÀH =2.4 Hz), 7.17–
7.23 (m, 3H), 7.36–7.40 (m, 1H), 7.45–7.50 (m, 3H), 7.57 (d, 2H, J=
8.0 Hz), 7.67 (d, 2H, J=8.0 Hz), 7.78 (s, 1H), 8.33 ppm (s, 1H);
13C NMR (100 MHz, [D6]DMSO): d=43.8, 56.1, 62.4, 63.2, 75.2,
103.5, 110.3, 126.4, 126.5 (2C), 126.7 (2C), 127.4, 129.1 (2C), 129.3
(2C), 130.1, 138.4, 138.9, 140.2, 144.8, 150.6 ppm (CF not visible); IR
(KBr): n˜ =3450, 1614, 1495, 1267, 1132 cmÀ1; MS (ESI) m/z (%):
435.5 (100) [M+H]+; UPLC purity 99%.
using the Biobyte program: http://biobyte.com.index.html) is
certainly of utmost importance for further development.
Experimental Section
Chemistry
General methods: Melting points were determined using an Elec-
trothermal IA9300 digital melting point apparatus and are uncor-
1
rected. H and 13C NMR spectra were recorded on a Bruker AC250
(250 MHz) or Bruker Avance 400 spectrometer (400 MHz). Chemi-
cal shifts are expressed as d values (ppm) relative to tetramethyl-
silane (TMS) as an internal standard (s=singlet, d=doublet, t=
triplet, q=quadruplet, sext=sextuplet, m=multiplet and b=
broad). Coupling constants (J) are given in Hertz (Hz). IR spectra
were obtained in KBr pellets using a Perkin–Elmer Paragon FTIR
1000 PC spectrometer. Only the most significant absorption bands
have been reported. Electrospray ionization (ESI) mass spectromet-
ric analysis was performed on a Waters Acquity UPLC System ZQ
2000 single quadrupole. All compounds tested displayed more
than 96% purity. All reactions were monitored by thin-layer chro-
matography (TLC) using 0.2 mm 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 were
commercially available. Focused microwave irradiations were car-
ried out with a CEM Discover focused microwave reactor (300 W,
2455 MHz, monomode system).
2-(2,4-Difluorophenyl)-1-{[4-(4-methoxyphenyl)benzyl]methyl-
amino}-3-(1H-,2,4-triazol-1-yl)propan-2-ol (5): The synthetic pro-
cedure for compound 4 was used, beginning from 2 (200 mg,
0.41 mmol)
and
4-methoxybenzeneboronic
acid
(75 mg,
0.50 mmol) to yield compound 5 as a white powder (145 mg,
75%): Rf =0.25 (CH2Cl2/EtOH 98:2); mp: 109–1108C; 1H NMR
(400 MHz, [D6]DMSO): d=2.12 (s, 3H), 2.81 (d, 1H, J=13.6 Hz),
3.07 (d, 1H, J=13.6 Hz), 3.46 (d, 1H, J=13.2 Hz), 3.61 (d, 1H, J=
13.2 Hz), 3.82 (s, 3H), 4.55 (d, 1H, J=14.4 Hz), 4.61 (d, 1H, J=
2-(2,4-Difluorophenyl)-1-[(4-iodobenzyl)methylamino]-3-(1H-
1,2,4-triazol-1-yl)propan-2-ol
(2):
N-N-Diisopropylethylamine
(774 mL, 4.47 mmol) was added to a stirred solution of 1 (1 g,
3.73 mmol) in CH3CN (31 mL), followed by addition of 4-iodoben-
zylbromide (1.107 g, 3.73 mmol). The solution was stirred at RT for
24 h. The solvent was removed in vacuo, and the residue was parti-
tioned between CH2Cl2 and H2O. The organic layers were dried
over anhyd Na2SO4 and concentrated in vacuo. The residue was
purified using silica gel column chromatography (CH2Cl2 and
CH2Cl2/EtOH, 98:2) to yield compound 2 as a white powder (1.67 g,
92%): Rf =0.30 (CH2Cl2/EtOH, 98:2); mp: 98–998C; 1H NMR
(400 MHz, [D6]DMSO): d=2.09 (s, 3H), 2.77 (d, 1H, J=13.7 Hz),
3.04 (d, 1H, J=13.7 Hz), 3.38 (d, 1H, J=13.5 Hz), 3.55 (d, 1H, J=
13.5 Hz), 4.53 (d, 1H, J=14.3 Hz), 4.58 (d, 1H, J=14.3 Hz), 5.78 (s,
1H), 6.92 (d, 2H, J=8.0 Hz), 6.99 (ddd, 1H, JHÀF =JHÀH =8.4 Hz,
14.4 Hz), 5.79 (s, 1H), 6.99 (ddd, 1H, JHÀF =JHÀH =8.4 Hz, JHÀH
=
2.4 Hz), 7.04 (d, 2H, J=8.4 Hz), 7.16 (d, 2H, J=8.0 Hz), 7.20 (ddd,
1H, JHÀF =J’HÀF =9.2 Hz, JHÀH =2.4 Hz), 7.48 (ddd, 1H, JHÀH =8.4 Hz,
J
HÀF =J’HÀF =6.8 Hz), 7.51 (d, 2H, J=8.0 Hz), 7.61 (d, 2H, J=8.4 Hz),
7.78 (s, 1H), 8.33 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=
43.7, 55.3, 56.1, 62.5, 63.2, 75.2, 103.9, 110.8, 114.5 (2C), 126.0 (2C),
126.4, 127.8 (2C), 129.3 (2C), 130.0, 132.5, 137.6, 138.6, 145.1,
150.6, 159.0 ppm (CF not visible); IR (KBr): n˜ =3450, 1612, 1497,
1260, 1123 cmÀ1; MS (ESI) m/z (%): 82.9 (35), 197.1 (24), 465.1 (100)
[M+H]+; UPLC purity 98%.
J
HÀH =2.4 Hz), 7.18 (ddd, 1H, JHÀF =J’HÀF =9.2 Hz, JHÀH =2.4 Hz), 7.44
1-{[4-(4-Cyanophenyl)benzyl]methylamino}-2-(2,4-difluorophen-
yl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (6): To a 10 mL vial were
added under argon 2 (200 mg, 0.41 mmol), 4-cyanobenzeneboron-
ic acid (61 mg, 0.41 mmol), Pd(PPh3)4 (24 mg, 0.02 mmol), 2m aq
Na2CO3 (2 mL) and acetonitrile (2 mL). The resulting mixture was
heated at 1208C under microwave irradiation (100 W) for 10 min.
The reaction mixture was cooled to RT and diluted with H2O. Prod-
uct was extracted with CH2Cl2, organic layers were dried over
anhyd Na2SO4 and concentrated in vacuo. The residue was purified
on silica gel column chromatography (CH2Cl2 and CH2Cl2/EtOH,
99:1) to yield compound 6 as a white powder (123 mg, 65%): Rf =
0.30 (CH2Cl2/EtOH, 98:2); mp: 132–1348C; 1H NMR (400 MHz,
[D6]DMSO): d=2.13 (s, 3H), 2.81 (d, 1H, J=13.6 Hz), 3.09 (d, 1H,
J=13.6 Hz), 3.49 (d, 1H, J=13.2 Hz), 3.66 (d, 1H, J=13.2 Hz), 4.55
(d, 1H, J=14.0 Hz), 4.61 (d, 1H, J=14.0 Hz), 5.81 (s, 1H), 7.01 (ddd,
(ddd, 1H, JHÀH =8.4 Hz, JHÀF =J’HÀF =6.8 Hz), 7.62 (d, 2H, J=8.0 Hz),
7.78 (s, 1H), 8.31 ppm (s, 1H); IR (KBr): n˜ =3451, 1613, 1495, 1272,
1127 cmÀ1; MS (ESI) m/z (%): 485.0 (100) [M+H]+.
2-(2,4-Difluorophenyl)-1-[(3-iodobenzyl)methylamino]-3-(1H-
1,2,4-triazol-1-yl)propan-2-ol (3): The synthetic procedure for
compound 2 was used, beginning from 1 (1.0 g, 3.73 mmol) and 3-
iodobenzylbromide (1.1 mg, 3.73 mmol), to yield compound 3 as a
white powder (975 mg, 54%): Rf =0.15 (CH2Cl2/EtOH, 98:2); mp:
85–868C; 1H NMR (400 MHz, [D6]DMSO): d=2.08 (s, 3H), 2.73 (d,
1H, J=13.7 Hz), 3.10 (d, 1H, J=13.7 Hz), 3.30 (d, 1H, J=13.5 Hz),
3.61 (d, 1H, J=13.5 Hz), 4.52 (d, 1H, J=14.2 Hz), 4.57 (d, 1H, J=
14.2 Hz), 5.84 (s, 1H), 7.00 (ddd, 1H, JHÀF =JHÀH =8.4 Hz, JHÀH
2.4 Hz), 7.04–7.12 (m, 2H), 7.19 (ddd, 1H, JHÀF =J’HÀF =9.2 Hz,
HÀH =2.4 Hz), 7.29 (s, 1H), 7.46 (ddd, 1H, JHÀH =8.4 Hz, JHÀF
=
J
=
1H, JHÀF =JHÀH =8.4 Hz, JHÀH =2.4 Hz), 7.20 (dd, 1H, JHÀF =J’HÀF
9.2 Hz), 7.24 (d, 2H, J=8.0 Hz), 7.48 (ddd, 1H, JHÀH =8.4 Hz, JHÀF
=
=
J’HÀF =6.8 Hz), 7.57 (d, 1H, J=7.6 Hz), 7.80 (s, 1H), 8.32 ppm (s,
1H); IR (KBr): n˜ =3428, 3020, 1613, 1498, 1415, 1277, 1141 cmÀ1
MS (ESI) m/z (%): 485.0 (100) [M+H]+.
;
J’HÀF =6.8 Hz), 7.67 (d, 2H, J=8.0 Hz), 7.79 (s, 1H), 7.90 (d, 2H, J=
7.6 Hz), 7.95 (d, 2H, J=7.6 Hz), 8.32 ppm (s, 1H); 13C NMR
(100 MHz, [D6]DMSO): d=43.6, 55.9, 62.1, 63.0, 75.1, 103.7, 109.7,
110.5, 118.9, 126.7 (2C), 127.3 (2C), 128.0, 129.3 (2C), 130.0, 132.8
2-(2,4-Difluorophenyl)-1-[(biphenyl-4-ylmethyl)methylamino]-3-
(1H-1,2,4-triazol-1-yl)propan-2-ol
(4):
Pd(PPh3)4
(14 mg,
ChemMedChem 2011, 6, 1806 – 1815
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemmedchem.org
1811

C. Logꢀ et al.
MED
(2C), 136.7, 139.8, 144.4, 144.9, 150.4 ppm (CF not visible); IR (KBr):
n˜ =3451, 2360, 1615, 1503, 1272, 1108 cmÀ1; MS (ESI) m/z (%):
460.1 (100) [M+H]+; UPLC purity 98%.
[D6]DMSO): d=2.13 (s, 3H), 2.81 (d, 1H, J=14.5 Hz), 3.10 (d, 1H,
J=14.5 Hz), 3.49 (d, 1H, J=13.6 Hz), 3.67 (d, 1H, J=13.6 Hz), 4.55
(d, 1H, J=14.2 Hz), 4.61 (d, 1H, J=14.2 Hz), 5.81 (s, 1H), 7.02 (dd,
1H, JHÀF =JHÀH =8.4 Hz), 7.19 (ddd, 1H, JHÀF =J’HÀF =9.2 Hz, JHÀH
2.4 Hz), 7.25 (d, 2H, J=8.4 Hz), 7.48 (ddd, 1H, JHÀH =8.4 Hz, JHÀF
=
=
2-(2,4-Difluorophenyl)-1-{[4-(4-nitrophenyl)benzyl]methylamino}-
3-(1H-1,2,4-triazol-1-yl)propan-2-ol (7): The synthetic procedure
for compound 6 was used, beginning from 2 (200 mg, 0.41 mmol)
and 4-nitrobenzeneboronic acid (68 mg, 0.41 mmol), to yield com-
pound 7 as a yellow powder (123 mg, 62%): Rf =0.20 (CH2Cl2/EtOH,
99:1); mp: 125–1268C; 1H NMR (400 MHz, [D6]DMSO): d=2.12 (s,
3H), 2.81 (d, 1H, J=14.0 Hz), 3.10 (d, 1H, J=14.0 Hz), 3.50 (d, 1H,
J=13.6 Hz), 3.68 (d, 1H, J=13.6 Hz), 4.55 (d, 1H, J=14.4 Hz), 4.61
(d, 1H, J=14.4 Hz), 5.81 (s, 1H), 7.02 (ddd, 1H, JHÀF =JHÀH =8.4 Hz,
J’HÀF =6.8 Hz), 7.72 (m, 4H), 7.79 (s, 1H), 8.33 (s, 1H), 8.65 ppm (d,
2H, J=9.6 Hz); 13C NMR (100 MHz, [D6]DMSO): d=43.8, 56.1, 62.4,
63.2, 75.4, 104.6, 111.0, 118.0 (2C), 126.7 (2C), 127.0, 129.5 (2C),
130.4, 135.8, 140.6, 145.1, 146.9, 150.4 (2C), 150.7 ppm (CF not visi-
ble); IR (KBr): n˜ =3447, 1626, 1497, 1267, 1112 cmÀ1; MS (ESI) m/z
(%): 69.6 (18), 175.1 (100), 195.6 (33), 436.1 (58) [M+H]+; UPLC
purity 99%.
J
J
HÀH =2.4 Hz), 7.20 (m, 1H), 7.26 (d, 2H, J=8.0 Hz), 7.48 (ddd, 1H,
HÀH =8.4 Hz, JHÀF =J’HÀF =6.8 Hz), 7.71 (d, 2H, J=8.8 Hz), 7.79 (s,
2-(2,4-Difluorophenyl)-1-{[4-(1-methylpyrazol-4-yl)benzyl]meth-
ylamino}-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (11): The synthetic
procedure for compound 6 was used, beginning from 2 (200 mg,
0.41 mmol) and 1-methylpyrazole-4-boronic acid pinacol ester
(86 mg, 0.41 mmol) to yield compound 11 as a white powder
1H), 7.99 (d, 2H, J=8.8 Hz), 8.32 (s, 1H), 8.33 ppm (d, 2H, J=
8.8 Hz); 13C NMR (100 MHz, [D6]DMSO): d=43.8, 56.1, 62.3, 63.2,
75.3, 104.1, 110.9, 124.3 (2C), 126.0, 127.1 (2C), 127.8 (2C), 129.5
(2C), 130.3, 137.3, 140.4, 145.1, 146.6, 148.3, 150.7 ppm (CF not visi-
ble); IR (KBr): n˜ =3442, 1632, 1508,1496, 1338 cmÀ1; MS (ESI) m/z
(%): 480.1 (100) [M+H]+; UPLC purity 96%.
1
(97 mg, 53%): Rf =0.10 (CH2Cl2/EtOH, 98:2); mp: 84–858C; H NMR
(400 MHz, [D6]DMSO): d=2.11 (s, 3H), 2.79 (d, 1H, J=13.6 Hz),
3.05 (d, 1H, J=13.6 Hz), 3.40 (d, 1H, J=13.2 Hz), 3.56 (d, 1H, J=
13.2 Hz), 3.88 (s, 3H), 4.53 (d, 1H, J=14.2 Hz), 4.59 (d, 1H, J=
14.2 Hz), 5.78 (s, 1H), 7.00 (dd, 1H, JHÀF =JHÀH =8.4 Hz), 7.07 (d, 2H,
J=8.0 Hz), 7.19 (dd, 1H, JHÀF =J’HÀF =9.2 Hz), 7.44–7.47 (m, 3H),
7.78 (s, 1H), 7.85 (s, 1H), 8.12 (s, 1H), 8.32 ppm (s, 1H); 13C NMR
(100 MHz, [D6]DMSO): d=38.8, 43.8, 56.1, 62.6, 63.1, 75.3, 103.0,
110.9, 122.0, 124.8 (2C), 126.5, 127.8, 129.3 (2C), 130.1, 131.4,
136.1, 136.7, 145.1, 150.6 ppm (CF not visible); IR (KBr): n˜ =3448,
1630, 1497, 1272, 1133 cmÀ1; MS (ESI) m/z (%): 82.9 (100), 171.1
(24), 439.1 (73) [M+H]+; UPLC purity 98%.
1-{[4-(4-Trifluoromethylphenyl)benzyl]methylamino}-2-(2,4-di-
fluorophenyl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (8): The syn-
thetic procedure for compound 6 was used, beginning from 2
(200 mg, 0.41 mmol) and 4-trifluoromethylbenzeneboronic acid
(78 mg, 0.41 mmol), to yield compound 8 as a white powder
(135 mg, 65%): Rf =0.20 (CH2Cl2/EtOH, 98:2); mp: 92–938C; 1H NMR
(400 MHz, [D6]DMSO): d=2.13 (s, 3H), 2.81 (d, 1H, J=13.9 Hz),
3.10 (d, 1H, J=13.9 Hz), 3.49 (d, 1H, J=13.1 Hz), 3.66 (d, 1H, J=
13.1 Hz), 4.55 (d, 1H, J=13.8 Hz), 4.61 (d, 1H, J=13.8 Hz), 5.81 (s,
1H), 7.01 (ddd, 1H, JHÀF =JHÀH =8.4 Hz, JHÀH =2.4 Hz), 7.18–7.25 (m,
3H), 7.47 (ddd, 1H, JHÀH =8.4 Hz, JHÀF =J’HÀF =6.8 Hz), 7.65 (d, 2H,
J=6.8 Hz), 7.79 (s, 1H), 7.83 (d, 2H, J=7.2 Hz), 7.91 (d, 2H, J=
7.2 Hz), 8.33 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=43.8,
56.1, 62.4, 63.2, 75.3, 103.9, 110.9, 125.8, 125.9 (2C), 126.4, 126.9
(2C), 127.5 (2C), 127.9, 129.5 (2C), 130.0, 137.3, 139.7, 144.1, 145.1,
150.7 ppm (CF not visible); IR (KBr): n˜ =3452, 1626, 1497,
1123 cmÀ1; MS (ESI) m/z (%): 503.1 (100) [M+H]+; UPLC purity
98%.
2-(2,4-Difluorophenyl)-1-[(biphenyl-3-ylmethyl)methylamino]-3-
(1H-1,2,4-triazol-1-yl)propan-2-ol (12): Using the synthetic proce-
dure used for compound 4 starting from 3 (200 mg, 0.41 mmol)
and benzeneboronic acid (60 mg, 0.50 mmol) to yield compound
12 as a white powder (100 mg, 56%): Rf =0.20 (CH2Cl2/EtOH, 98:2);
1
mp: 74–758C; H NMR (400 MHz, [D6]DMSO): d=2.11 (s, 3H), 2.79
(d, 1H, J=13.6 Hz), 3.16 (d, 1H, J=13.6 Hz), 3.43 (d, 1H, J=
13.2 Hz), 3.72 (d, 1H, J=13.2 Hz), 4.53 (d, 1H, J=14.0 Hz), 4.58 (d,
1H, J=14.0 Hz), 5.84 (s, 1H), 7.00 (ddd, 1H, JHÀF =JHÀH =8.4 Hz,
J
HÀH =2.4 Hz), 7.10 (d, 1H, J=7.6 Hz), 7.17 (ddd, 1H, JHÀF =J’HÀF =
2-(2,4-Difluorophenyl)-1-{[4-(pyridin-3-yl)benzyl]methylamino}-3-
(1H-1,2,4-triazol-1-yl)propan-2-ol (9): The synthetic procedure for
compound 6 was used, beginning from 2 (200 mg, 0.41 mmol) and
pyridine-3-boronic acid (51 mg, 0.41 mmol), to yield compound 9
as a white powder (73 mg, 41%): Rf =0.05 (CH2Cl2/EtOH, 98:2); mp:
87–888C; 1H NMR (400 MHz, [D6]DMSO): d=2.09 (s, 3H), 2.77 (d,
1H, J=14.0 Hz), 3.05 (d, 1H, J=14.0 Hz), 3.45 (d, 1H, J=13.2 Hz),
3.61 (d, 1H, J=13.2 Hz), 4.51 (d, 1H, J=14.0 Hz), 4.57 (d, 1H, J=
14.0 Hz), 5.77 (s, 1H), 6.97 (dd, 1H, JHÀF =JHÀH =8.4 Hz), 7.15 (m,
1H), 7.19 (d, 2H, J=7.6 Hz), 7.45 (m, 1H), 7.46 (m, 1H), 7.60 (d, 2H,
J=7.6 Hz), 7.75 (s, 1H), 8.04 (d, 1H, J=7.2 Hz), 8.29 (s, 1H), 8.54
(m, 1H), 8.86 ppm (m, 1H); 13C NMR (100 MHz, [D6]DMSO): d=
43.8, 56.0, 62.4, 63.2, 75.3, 103.5, 110.3, 124.0, 126.7 (2C), 127.6,
129.5 (2C), 130.1, 134.1, 135.5, 135.8, 139.3, 145.1, 147.2, 148.5,
150.6 ppm (CF not visible); IR (KBr): n˜ =3452, 1638, 1502, 1272,
1107 cmÀ1; MS (ESI) m/z (%): 82.9 (100), 175.1 (3) 436.2 (5) [M+H]+;
UPLC purity 97%.
9.2 Hz, JHÀH =2.4 Hz), 7.26 (s, 1H), 7.36 (dd, 1H, J=7.6 Hz), 7.40 (d,
1H, J=7.6 Hz), 7.34–7.60 (m, 6H), 7.79 (s, 1H), 8.32 ppm (s, 1H);
13C NMR (100 MHz, [D6]DMSO): d=43.7, 56.2, 62.6 63.3, 75.5, 103.8,
110.8, 125.4, 126.4, 126.8 (2C), 126.9, 127.5, 127.9, 128.8, 129.0
(2C), 130.1, 140.0, 140.1, 140.3, 145.1, 150.6, 158.9, 161.6 ppm; IR
(KBr): n˜ =3446, 1618, 1501, 1267, 1133 cmÀ1; MS (ESI) m/z (%):
435.2 (100) [M+H]+; UPLC purity 100%.
1-{[3-(4-Cyanophenyl)benzyl]methylamino}2-(2,4-difluorophen-
yl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (13): The synthetic proce-
dure for compound 6 was used, beginning from 3 (200 mg,
0.41 mmol) and 4-cyanobenzeneboronic acid (61 mg, 0.41 mmol)
to yield compound 13 as a white powder (135 mg, 70%): Rf =0.20
1
(CH2Cl2/EtOH, 98:2); mp: 45–468C; H NMR (400 MHz, [D6]DMSO):
d=2.12 (s, 3H), 2.78 (d, 1H, J=13.6 Hz), 3.16 (d, 1H, J=13.6 Hz),
3.44 (d, 1H, J=13.4 Hz), 3.73 (d, 1H, J=13.4 Hz), 4.52 (d, 1H, J=
14.1 Hz), 4.58 (d, 1H, J=14.1 Hz), 5.85 (s, 1H), 7.00 (m, 1H), 7.14–
7.19 (m, 2H), 7.31 (s, 1H), 7.41 (t, 1H, J=7.6 Hz), 7.48 (m, 1H), 7.60
(d, 1H, J=7.6 Hz), 7.78–7.80 (m, 3H), 7.95 (d, 2H, J=8.0 Hz),
8.32 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=43.8, 56.2,
62.4, 63.2, 75.5, 103.9, 110.1, 110.8, 119.1, 125.7, 126.4, 127.2, 127.7
(2C), 129.0, 129.1, 130.0, 133.0 (2C), 138.2, 140.4, 144.8, 145.1,
150.6, 159.2, 161.8 ppm; IR (KBr): n˜ =3457, 2216, 1618, 1497, 1272,
2-(2,4-Difluorophenyl)-1-{[4-(pyridin-4-yl)benzyl]methylamino}-3-
(1H-1,2,4-triazol-1-yl)propan-2-ol (10): The synthetic procedure
for compound 6 was used, beginning from 2 (200 mg, 0.41 mmol)
and pyridine-4-boronic acid pinacol ester (85 mg, 0.41 mmol), to
yield compound 10 as a light brown powder (95 mg, 53%): Rf =
0.03 (CH2Cl2/EtOH, 98:2); mp: 104–1058C; 1H NMR (400 MHz,
1812
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ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
ChemMedChem 2011, 6, 1806 – 1815

Antifungal Agents
1133 cmÀ1; MS (ESI) m/z (%): 460.2 (100) [M+H]+; UPLC purity
100%.
8.4 Hz), 7.16 (d, 2H, J=8.0 Hz), 7.20 (m, 1H), 7.47 (m, 1H), 7.52 (d,
2H, J=8.0 Hz), 7.78 (s, 1H), 7.86 (dd, 1H, J=8.8 Hz, J=1.6 Hz), 8.33
(s, 1H), 8.46 ppm (d, 1H, J=1.6 Hz); 13C NMR (100 MHz,
[D6]DMSO): d=43.7, 44.8 (2C), 46.0, 54.5 (2C), 55.2, 62.5, 63.1, 75.2,
101.5, 107.2, 111.3, 124.5, 125.5 (2C), 126.4, 129.4 (2C), 129.0, 135.8,
136.3, 137.5, 145.1, 145.5, 150.6, 158.4 ppm (CF not visible); IR
(KBr): n˜ =3445, 1622, 1492, 1246, 1133 cmÀ1; MS (ESI) m/z (%):
195.6 (50), 224.1(85), 244.6 (50), 267.7 (100), 288.2 (54), 534.2 (16)
[M+H]+; UPLC purity 98%.
2-(2,4-Difluorophenyl)-1-{[3-(pyridin-3-yl)benzyl]methylamino}-3-
(1H-1,2,4-triazol-1-yl)propan-2-ol (14): The synthetic procedure
for compound 6 was used, beginning from 3 (200 mg, 0.41 mmol)
and pyridine-3-boronic acid (51 mg, 0.41 mmol), to yield com-
pound 14 as a colorless oil (100 mg, 56%): Rf =0.45 (CH2Cl2/EtOH,
1
95:5); H NMR (400 MHz, [D6]DMSO): d=2.07 (s, 3H), 2.74 (d, 1H,
J=13.6 Hz), 3.09 (d, 1H, J=13.6 Hz), 3.41 (d, 1H, J=13.6 Hz), 3.67
(d, 1H, J=13.6 Hz), 4.48 (d, 1H, J=14.0 Hz), 4.53 (d, 1H, J=
2-(2,4-Difluorophenyl)-1-{[4-(6-morpholin-4-ylpyridin-3-yl)benz-
yl]methylamino}-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (17): The
synthetic procedure for compound 16 was used, beginning from
15 (150 mg, 0.29 mmol) and morpholine (30 mL, 0.35 mmol), to
yield compound 17 as a yellow powder (110 mg, 72%): Rf =0.30
(CH2Cl2/EtOH, 90:10); mp: 150–1518C; 1H NMR (400 MHz,
[D6]DMSO): d=2.12 (s, 3H), 2.81 (d, 1H, J=13.6 Hz), 3.08 (d, 1H,
J=13.6 Hz), 3.45 (d, 1H, J=13.2 Hz), 3.52 (t, 4H, J=4.4 Hz), 3.61 (d,
1H, J=13.2 Hz), 3.75 (t, 4H, J=4.0 Hz), 4.55 (d, 1H, J=14.0 Hz),
4.60 (d, 1H, J=14.0 Hz), 5.79 (s, 1H), 6.94 (d, 1H, J=8.4 Hz), 7.01
(ddd, 1H, JHÀF =JHÀH =8.4 Hz), 7.16 (d, 2H, J=8.0 Hz), 7.20 (dd, 1H,
14.0 Hz), 5.79 (s, 1H), 6.94 (ddd, 1H, JHÀF =JHÀH =8.4 Hz, JHÀH
=
2.4 Hz), 7.08–7.12 (m, 2H), 7.29 (s, 1H), 7.35 (t, 1H, J=7.6 Hz), 7.40–
7.49 (m, 2H), 7.54 (d, 1H, J=7.6 Hz), 7.73 (s, 1H), 7.93 (dd, 1H, J=
7.6 Hz, J=1.2 Hz), 8.27 (s, 1H), 8.56 (d, 1H, J=7.6 Hz), 8.80 ppm (s,
1H); 13C NMR (100 MHz, [D6]DMSO): d=43.8, 56.2, 62.5, 63.2, 75.4,
103.8, 110.7, 124.0, 125.6, 126.4, 127.1, 128.5, 129.0, 130.0, 134.2,
135.7, 137.1, 140.3, 145.1, 147.8, 148.6, 150.6, 159.1, 161.8 ppm; IR
(NaCl): n˜ =3411, 1610, 1497, 1456, 1272, 1133 cmÀ1; MS (ESI)
m/z (%): 69.9 (30), 175.2 (36), 195.7 (100), 239.2 (46), 436.2 (69)
[M+H]+; UPLC purity 98%.
J
HÀF =J’HÀF =9.6 Hz), 7.47 (m, 1H), 7.53 (d, 2H, J=8.0 Hz), 7.78 (s,
1-{[4-(6-Bromopyridin-3-yl)benzyl]methylamino}-2-(2,4-difluoro-
1H), 7.89 (dd, 1H, J=8.4 Hz, J=1.6 Hz), 8.33 (s, 1H), 8.48 ppm (d,
1H, J=1.6 Hz); 13C NMR (100 MHz, [D6]DMSO): d=43.7, 45.3 (2C),
56.4, 62.5, 63.4, 66.1 (2C), 75.3, 104.1, 107.1, 110.8, 124.5, 125.5
(2C), 127.0, 129.4 (2C), 130.3, 134.3, 135.3, 137.0, 145.1, 145.5,
150.6, 158.4 ppm (CF not visible); IR (KBr): n˜ =3453, 1626, 1482,
1231, 1108 cmÀ1; MS (ESI) m/z (%): 82.8 (78), 217.6 (100), 261.1 (50),
521.2 (12) [M+H]+; UPLC purity 96%.
phenyl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol
(15):
Pd(PPh3)4
(378 mg, 0.33 mmol) was added under argon to a stirred solution
of 2 (2.29 g, 4.73 mmol) in DMF (67 mL), and the solution was
stirred at RT for 20 min. 2-Bromopyridine-5-boronic acid (1.5 g,
7.43 mmol) was added, followed by the addition of 1m aq Na2CO3
(23.7 mL, 23.70 mmol). The resulting mixture was heated at 908C
for 2 h, then cooled to RT and diluted with H2O. The product was
extracted with EtOAc, and the organic layers were dried over
anhyd Na2SO4 and concentrated in vacuo. The residue was purified
using silica gel column chromatography (CH2Cl2 and CH2Cl2/EtOH,
99:1) to yield compound 15 as a white powder (1.467 g, 60%): Rf =
0.25 (CH2Cl2/EtOH, 98:2); mp: 127–1288C; 1H NMR (400 MHz,
[D6]DMSO): d=2.12 (s, 3H), 2.80 (d, 1H, J=13.5 Hz), 3.10 (d, 1H,
J=13.5 Hz), 3.48 (d, 1H, J=13.5 Hz), 3.64 (d, 1H, J=13.5 Hz), 4.55
(d, 1H, J=14.3 Hz), 4.60 (d, 1H, J=14.3 Hz), 5.81 (s, 1H), 7.02 (dd,
1H, JHÀF =JHÀH =8.8 Hz), 7.19 (m, 1H), 7.23 (d, 2H, J=8.0 Hz), 7.49
(ddd, 1H, JHÀH =8.8 Hz, JHÀF =J’HÀF =6.8 Hz), 7.65 (d, 2H, J=8.0 Hz),
7.75 (d, 1H, J=8.4 Hz), 7.79 (s, 1H), 8.07 (dd, 1H, J=8.4 Hz, J=
2.0 Hz), 8.33 (s, 1H), 8.73 ppm (d, 1H, J=2.0 Hz); 13C NMR
(100 MHz, [D6]DMSO): d=43.8, 46.5, 62.3, 62.9, 75.3, 103.5, 110.4,
126.7 (2C), 128.0, 128.2, 129.5 (2C), 130.0, 134.3, 135.3, 137.0,
137.6, 139.8, 145.1, 148.4, 150.7, 154.9, 157.8 ppm; IR (KBr): n˜ =
3484, 1641, 1497, 1272, 1082 cmÀ1; MS (ESI) m/z (%): 514.0 (47),
516.0 (50) [M+H]+; UPLC purity 97%.
2-(2,4-Difluorophenyl)-1-({4-[6-(diphenylmethylideneamino)-pyri-
din-3-yl]benzyl}methylamino)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol
(18): The synthetic procedure for compound 16 was used, begin-
ning from 15 (100 mg, 0.19 mmol) and benzophenone imine
(39 mL, 0.23 mmol), to yield compound 18 as a yellow powder
1
(83 mg, 70%): Rf =0.10 (CH2Cl2/EtOH, 98:2); mp: 71–728C; H NMR
(400 MHz, [D6]DMSO): d=2.10 (s, 3H), 2.79 (d, 1H, J=13.5 Hz),
3.07 (d, 1H, J=13.5 Hz), 3.45 (d, 1H, J=13.2 Hz), 3.62 (d, 1H, J=
13.2 Hz), 4.54 (d, 1H, J=14.4 Hz), 4.60 (d, 1H, J=14.4 Hz), 5.79 (s,
1H), 6.79 (d, 1H, J=8.4 Hz), 7.00 (ddd, 1H, JHÀF =JHÀH =8.4 Hz,
J
HÀH =2.4 Hz), 7.19 (m, 5H), 7.36 (m, 2H), 7.44 (ddd, 1H, JHÀH =
8.4 Hz, JHÀF =J’HÀF =6.8 Hz), 7.59 (m, 6H), 7.74 (m, 2H), 7.78 (s, 1H),
7.79 (dd, 1H, J=8.4 Hz, J=1.6 Hz), 8.33 (s, 1H), 8.59 ppm (d, 1H,
J=1.6 Hz); IR (KBr): n˜ =3448, 1630, 1497, 1272 cmÀ1
.
1-{[4-(6-Aminopyridin-3-yl)benzyl]methylamino}-2-(2,4-difluoro-
phenyl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (19): Sodium acetate
(255 mg, 3.11 mmol) was added under argon to a strirred solution
of 18 (210 mg, 0.34 mmol) in MeOH (21 mL), followed by the addi-
tion of hydroxylamine hydrochloride (163 mg, 2.34 mmol). The so-
lution was stirred at RT for 2 h. The solvent was removed in vacuo
and the residue was diluted with CH2Cl2. Organic layers were
washed with 0.1m aq NaOH, dried over anhyd Na2SO4, and concen-
trated in vacuo. The residue was triturated in Et2O to yield com-
pound 19 after filtration as a white powder (127 mg, 82%): Rf =
0.45 (CH2Cl2/EtOH, 98:2); mp: 151–1528C; 1H NMR (400 MHz,
[D6]DMSO): d=2.11 (s, 3H), 2.80 (d, 1H, J=13.6 Hz), 3.06 (d, 1H,
J=13.6 Hz), 3.45 (d, 1H, J=13.6 Hz), 3.59 (d, 1H, J=13.6 Hz), 4.56
(d, 1H, J=14.4 Hz), 4.58 (d, 1H, J=14.4 Hz), 5.78 (s, 1H), 6.06 (s,
2H), 6.54 (d, 1H, J=8.8 Hz), 7.01 (dd, 1H, JHÀF =JHÀH =8.8 Hz), 7.13
(d, 2H, J=8.0 Hz), 7.19 (dd, 1H, JHÀF =J’HÀF =9.2 Hz), 7.46 (d, 2H,
J=8.0 Hz), 7.47 (m, 1H), 7.70 (dd, 1H, J=8.8 Hz, J=2.0 Hz), 7.78 (s,
1H), 8.24 (d, 1H, J=2.0 Hz), 8.33 ppm (s, 1H); 13C NMR (100 MHz,
[D6]DMSO): d=43.7, 56.1, 62.5, 63.2, 75.2, 103.9, 108.1, 110.8, 123.9,
125.2 (2C), 126.4, 129.4 (2C), 130.1, 135.4, 136.9, 137.1, 145.1,
2-(2,4-Difluorophenyl)-1-({4-[6-(4-methylpiperazin-1-yl)pyridin-3-
yl]benzyl)}methylamino)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (16):
Compound 15 (150 mg, 0.29 mmol), Pd2(dba)3 (5.4 mg,
0.006 mmol), (Æ)-BINAP (7.3 mg, 0.012 mmol), tBuONa (39 mg,
0.41 mmol), N-methylpiperazine (38 mL, 0.35 mmol) and toluene
(2.6 mL) were added to a 10 mL vial under argon. The resulting
mixture was heated at 1008C for 2 h. The reaction mixture was
cooled to RT and diluted with H2O. The product was extracted with
CH2Cl2, and the organic layers were dried over anhyd Na2SO4 and
concentrated in vacuo. The residue was purified using silica gel
column chromatography (CH2Cl2 and CH2Cl2/EtOH, 99:1) to yield
compound 16 as a white powder (128 mg, 82%): Rf =0.45 (CH2Cl2/
EtOH, 90:10); mp: 112–1138C; 1H NMR (400 MHz, [D6]DMSO): d=
2.12 (s, 3H), 2.27 (s, 3H), 2.45 (s, 4H), 2.81 (d, 1H, J=13.6 Hz), 3.08
(d, 1H, J=13.6 Hz), 3.45 (d, 1H, J=14.4 Hz), 3.56 (s, 4H), 3.61 (d,
1H, J=14.4 Hz), 4.55 (d, 1H, J=14.8 Hz), 4.60 (d, 1H, J=14.8 Hz),
5.80 (s, 1H), 6.93 (d, 1H, J=8.8 Hz), 7.01 (ddd, 1H, JHÀF =JHÀH
=
ChemMedChem 2011, 6, 1806 – 1815
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemmedchem.org
1813

C. Logꢀ et al.
MED
145.8, 150.6, 159.3 ppm (CF not visible); IR (KBr): n˜ =3451, 1638,
1492, 1272, 1133 cmÀ1; MS (ESI) m/z (%): 182.6 (100), 203.1 (43),
226.1 (7), 451.1 (9) [M+H]+; UPLC purity 100%.
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 while stirring. Compound 6 was
introduced into culture medium before incubation. Cells were col-
lected by centrifugation at 1500 g. The pellet was suspended in
3 mL of saponification medium (25 g KOH, 36 mL of distilled water
and brought to 100 mL with 100% ethanol). Then suspension was
mixed by vortex for 1 min and incubated at 808C for 120 min. Ster-
ols were then extracted by addition of 4 mL n-hexane (Merck,
Darmstadt, Germany), and the hexane extract was evaporated.
Samples were derivatized with 100 mL of silylating mixture (Fluka,
Saint Quentin Fallavier, France) at RT for 30 min, evaporated, and
diluted in 500 mL n-hexane. Aliquots of sample (2 mL) was injected
into a gas chromatograph (model 6890N, Agilent Technologies,
Palo Alto, CA, USA) coupled with a quadrupole mass spectrometer
(5973i, Agilent Technologies, Palo Alto, CA, USA). Analyses were
carried out in a splitless mode with helium as carrier gas (constant
rate of 1.2 mLminÀ1) at an injector temperature of 2508C. The
transfer line between gas chromatograph and mass spectrometer
was operated at 2908C, and the EI source was held at 2808C. The
GC capillary column was an HP-5MS (30 mꢃ0.25 mm ID, 0.25 mm
film thickness, Agilent Technologies). The GC oven was pro-
grammed as follows: initial temperature 1508C, hold for 0.5 min,
N-{5-[4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-
yl)propyl]methylamino}methyl)phenyl]pyridin-2-yl}methanesul-
fonamide (20): MsCl (42 mL, 0.53 mmol) was added under argon to
a stirred solution of 19 (100 mg, 0.22 mmol) in pyridine (3.4 mL),
and the solution was heated at 558C for 2 h. Solvent was removed
in vacuo, and the residue was diluted with H2O. The product was
extracted with CH2Cl2, and the organic layers were dried over
anhyd Na2SO4 and concentrated in vacuo. The residue was purified
using silica gel column chromatography (CH2Cl2 and CH2Cl2/EtOH,
99:1) to yield compound 20 as a white powder (86 mg, 73%): Rf =
0.40 (CH2Cl2/EtOH, 95:5); mp: 120–1218C; 1H NMR (400 MHz,
[D6]DMSO): d=2.12 (s, 3H), 2.81 (d, 1H, J=13.7 Hz), 3.09 (d, 1H,
J=13.7 Hz), 3.35 (s, 3H), 3.49 (d, 1H, J=13.3 Hz), 3.64 (d, 1H, J=
13.3 Hz), 4.55 (d, 1H, J=14.4 Hz), 4.60 (d, 1H, J=14.4 Hz), 5.79 (s,
1H), 7.01 (ddd, 1H, JHÀF =JHÀH =8.4 Hz, JHÀH =2.4 Hz), 7.09 (d, 1H,
J=8.8 Hz), 7.18 (m, 1H), 7.20 (d, 2H, J=8.0 Hz), 7.48 (ddd, 1H,
J
HÀH =8.8 Hz, JHÀF =J’HÀF =6.8 Hz), 7.58 (d, 2H, J=8.0 Hz), 7.78 (s,
1H), 8.07 (dd, 1H, J=8.8 Hz, J=2.4 Hz), 8.33 (s, 1H), 8.58 (d, 1H,
J=2.4 Hz), 10.70 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=
41.9, 43.8, 56.1, 62.4, 63.2, 75.2, 103.9, 110.9, 112.5, 126.2 (2C),
126.5, 127.0, 129.5 (2C), 130.1, 135.4, 137.1, 138.7, 145.1, 147.8,
150.6, 151.7 ppm (CF not visible); IR (KBr): n˜ =3457, 1646, 1497,
1374, 1272, 1124 cmÀ1; MS (ESI) m/z (%) 529.2 (100) [M+H]+; UPLC
purity 98%.
increase to 2808C at 408CminÀ1, increase to 3008C at 58CminÀ1
,
then hold for 6 min. Sterols of interest were identified by mass
spectrometry. In order to study the influence of treatment on
sterol abundance, the area under the curve (AUC) of each peak
was used to calculate a ratio: sterol AUC/sum of sterols AUC.
N-{5-[4-({[2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-
yl)propyl]methylamino}methyl)phenyl]pyridin-2-yl}benzenesul-
fonamide (21): The synthetic procedure for compound 20 was
used, beginning from 19 (115 mg, 0.26 mmol) and benzenesulfonyl
chloride (78 mL, 0.61 mmol), to yield compound 21 as a yellow
powder (115 mg, 76%): Rf =0.40 (CH2Cl2/EtOH, 95:5); mp: 105–
Molecular modeling
Molecular modeling studies were performed using Sybyl software
version 8.0[39] running on a Dell Precision T3400 workstation. The
structure of CYP51 from Mycobacterium tuberculosis complexed
with fluconazole (PDB code: 1A1)[40] was used as the template for
the homology model of CYP51–C. albicans using a previously de-
scribed protocol.[30,36] Flexible docking of azoles into the enzyme
active site was performed using GOLD software.[41] A distance con-
straint was applied from N4 of the triazole ring to the heme iron
(2.0< d <2.4 ꢂ). For each compound, the most stable docking
model was selected according to the best scored conformation as
predicted by the GoldScore scoring function.
1
1068C; H NMR (400 MHz, [D6]DMSO): d=2.05 (s, 3H), 2.74 (d, 1H,
J=14.0 Hz), 3.02 (d, 1H, J=14.0 Hz), 3.40 (d, 1H, J=13.6 Hz), 3.57
(d, 1H, J=13.6 Hz), 4.49 (d, 1H, J=14.0 Hz), 4.54 (d, 1H, J=
14.0 Hz), 5.74 (s, 1H), 6.96 (m, 1H), 7.11–7.30 (m, 4H), 7.44 (m, 1H),
7.48 (d, 2H, J=7.6 Hz), 7.54–7.62 (m, 3H), 7.73 (s, 1H), 7.91 (d, 2H,
J=7.6 Hz), 8.01 (d, 1H, d, J=8.8 Hz), 8.27 (s, 1H), 8.35 (s, 1H),
11.75 ppm (s, 1H); 13C NMR (100 MHz, [D6]DMSO): d=43.7, 56.0,
62.4 63.1, 75.2, 103.9, 110.8, 113.3, 125.7, 126.0 (2C), 126.3, 126.8
(2C), 129.2 (2C), 129.4 (2C), 130.0, 132.7, 134.7, 138.0, 138.8, 141.7,
145.1, 147.8, 151.8, 159.3, 159.4, 161.8 ppm; IR (KBr): n˜ =3443,
1640, 1605, 1500, 1370, 1274, 1142, 1089 cmÀ1; MS (ESI) m/z (%):
591.3 (100) [M+H]+; UPLC purity 100%.
Acknowledgements
We wish to express our thanks to Prof. Dominique Sanglard (In-
stitute of Microbiology, University of Lausanne, Switzerland) for
providing the DSY735 strain used in this study.
Biological assays
Antifungal activity: All of the compounds were screened for anti-
fungal activity against C. albicans CA98001, C. krusei (CK506, CK8),
C. glabrata (CG468), C. parapsilosis (CAPA1, CAPA2), and A. fumiga-
tus (AF98003) strains. Two C. albicans strains (DSY735, CAAL74)
known to have reduced susceptibility to fluconazole were also in-
vestigated.[31] DSY735 was a gift from Prof. D. Sanglard at the Insti-
tute of Microbiology (University of Lausanne, Switzerland). All
other strains were issued from our collection, and growth inhibi-
tion was measured as previously described.[37] Fluconazole, vorico-
nazole, and itraconazole were used as reference compounds. MIC
values were defined as the concentrations that inhibit growth of
Candida spp. or A. fumigatus by 50 or 80%, respectively, as recom-
mended by the Clinical and Laboratory Standards Institute M27 A3
Keywords: antifungal agents · azoles · Candida albicans ·
CYP51 inhibitors · structure–activity relationships
[1] B. C. Monk, A. Goffeau, Science 2008, 321, 367–369.
[2] P. Chandrasekar, J. Antimicrob. Chemother. 2011, 66, 457–465.
[3] ꢄ. Tꢅrel, Expert Rev. Anti-Infect. Ther. 2011, 9, 325–338.
[4] S. L. Kelly, A. Arnoldi, D. E. Kelly, Biochem. Soc. Trans. 1993, 21, 1034–
1038.
[5] S. L. Kelly, D. C. Lamb, D. E. Kelly, FEMS Microbiol. Lett. 1999, 180, 171–
175.
[6] S. L. Kelly, D. C. Lamb, J. Loeffler, H. Einsele, D. E. Kelly, Biochem. Biophys.
Res. Commun. 1999, 262, 174–179.
and M38 A2 guidelines.[38] MIC values are expressed in mgmLÀ1
.
1814
www.chemmedchem.org
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
ChemMedChem 2011, 6, 1806 – 1815

Antifungal Agents
[7] D. C. Lamb, D. E. Kelly, T. C. White, S. L. Kelly, Antimicrob. Agents Chemo-
ther. 2000, 44, 63–67.
[26] W. Wang, S. Wang, Y. Liu, G. Dong, Y. Cao, Z. Miao, J. Yao, W. Zhang, C.
Sheng, Eur. J. Med. Chem. 2010, 45, 6020–6026.
[8] G. Del Sorbo, H. Schoonbeek, M. A. De Waard, Fungal Genet. Biol. 2000,
30, 1–15.
[27] F. Giraud, C. Logꢀ, F. Pagniez, D. Crꢀpin, P. Le Pape, M. Le Borgne,
Bioorg. Med. Chem. Lett. 2008, 18, 1820–1824.
[9] J. Morschhꢆuser, Fungal Genet. Biol. 2010, 47, 94–106.
[10] A. S. Chau, M. Gurnani, R. Hawkinson, M. Laverdiere, A. Cacciapuoti,
P. M. McNicholas, Antimicrob. Agents Chemother. 2005, 49, 3646–3651.
[11] D. Sanglard, A. Coste, S. Ferrari, FEMS Yeast Res. 2009, 9, 1029–1050.
[12] J. L. Lopez-Ribot, R. K. McAtee, L. N. Lee, W. R. Kirkpatrick, T. C. White, D.
Sanglard, T. F. Patterson, Antimicrob. Agents Chemother. 1998, 42, 2932–
2937.
[13] A. G. S. Warrilow, N. Melo, C. M. Martel, J. E. Parker, W. D. Nes, S. L. Kelly,
D. E. Kelly, Antimicrob. Agents Chemother. 2010, 54, 4225–4234.
[14] A. G. S. Warrilow, C. M. Martel, J. E. Parker, N. Melo, D. C. Lamb, W. D.
Nes, D. E. Kelly, S. L. Kelly, Antimicrob. Agents Chemother. 2010, 54,
4235–4245.
[15] G. I. Lepesheva, H. W. Park, T. Y. Hargrove, B. Vanhollebeke, Z. Wawrzak,
J. M. Harp, M. Sundaramoorthy, W. D. Nes, E. Pays, M. Chaudhuri, F. Vil-
lalta, M. R. Waterman, J. Biol. Chem. 2010, 285, 1773–1780.
[16] C. K. Chen, S. S. F. Leung, C. Guilbert, M. Jacobson, J. H. Mckerrow, L. M.
Podust, Plos Negl. Trop. Dis. 2010, 4, e651.
[28] F. Giraud, R. Guillon, C. Logꢀ, F. Pagniez, C. Picot, M. Le Borgne, P.
Le Pape, Bioorg. Med. Chem. Lett. 2009, 19, 301–304.
[29] R. Guillon, F. Giraud, C. Logꢀ, M. Le Borgne, C. Picot, F. Pagniez, P.
Le Pape, Bioorg. Med. Chem. Lett. 2009, 19, 5833–5836.
[30] R. Guillon, C. Logꢀ, F. Pagniez, V. Ferchaud-Roucher, M. Duflos, C. Picot,
P. Le Pape, J. Enzyme Inhib. Med. Chem. 2011, 26, 261–269.
[31] R. Guillon, F. Pagniez, F. Giraud, D. Crꢀpin, C. Picot, M. Le Borgne, F.
Morio, M. Duflos, C. Logꢀ, P. Le Pape, ChemMedChem 2011, 6, 816–825.
[32] Y. Gong, W. He, Org. Lett. 2002, 4, 3803–3805.
[33] A. Bouillon, J. C. Lancelot, V. Collot, P. R. Bovy, S. Rault, Tetrahedron
2002, 58, 2885–2890.
[34] P. R. Parry, C. Wang, A. S. Batsanov, M. R. Bryce, B. Tarbit, J. Org. Chem.
2002, 67, 7541–7543.
[35] A. Lupetti, R. Danesi, M. Campa, M. D. Tacca, S. Kelly, Trends Mol. Med.
2002, 8, 76–81.
[36] F. Giraud, Ph. D. Thesis, Universitꢀ de Nantes, Nantes Atlantique Univer-
sitꢀs (France), 2007.
[17] L. Xiao, V. Madison, A. S. Chau, D. Loebenberg, R. E. Palermo, P. M. McNi-
cholas, Antimicrob. Agents Chemother. 2004, 48, 568–574.
[18] C. Sheng, W. Zhang, M. Zhang, Y. Song, H. Ji, J. Zhu, J. Yao, J. Yu, S.
Yang, Y. Zhou, J. Zhu, J. Lu, J. Biomol. Struct. Dyn. 2004, 22, 91–99.
[19] C. Sheng, Z. Miao, H. Ji, J. Yao, W. Wang, X. Che, G. Dong, J. Lu, W. Guo,
W. Zhang, Antimicrob. Agents Chemother. 2009, 53, 3487–3495.
[20] C. Sheng, W. Wang, X. Che, G. Dong, S. Wang, H. Ji, Z. Miao, J. Yao, W.
Zhang, ChemMedChem 2010, 5, 390–397.
[21] C. Sheng, W. Zhang, H. Ji, M. Zhang, Y. Song, H. Xu, J. Zhu, Z. Miao, Q.
Jiang, J. Yao, Y. Zhou, J. Zhu, J. Lꢅ, J. Med. Chem. 2006, 49, 2512–2525.
[22] C. Sheng, W. Zhang, Curr. Med. Chem. 2011, 18, 733–766.
[23] S. Chen, C. Sheng, H. Xu, Y. Jiang, W. Zhang, C. He, Biol. Pharm. Bull.
2007, 30, 1246–1253.
[37] F. Pagniez, P. Le Pape, J. Mycol. Med. 2001, 11, 73–78.
[38] Clinical and Laboratory Standards Institute a) Reference method for
broth dilution antifungal susceptibility testing of yeasts; Approved stan-
dard, 3rd ed, CLSI document M27-A3; b) Reference method for broth di-
lution antifungal susceptibility testing of filamentous fungi; Approved
standard, 2nd ed, CLSI document M38-A2, Clinical and Laboratory
Standards Institute, Wayne, PA 2008.
[39] Sybyl 8.0, Tripos International, 1699 South Hanley Rd., St. Louis, Missou-
ri, 63144, USA, 2007.
[40] L. M. Podust, T. L. Poulos, M. R. Waterman, Proc. Natl. Acad. Sci. USA
2001, 98, 3068–3073.
[41] G. Jones, P. Willet, R. C. Glen, J. Mol. Biol. 1997, 267, 727–748.
[24] C. Sheng, S. Chen, H. Ji, G. Dong, X. Che, W. Wang, Z. Miao, J. Yao, J. Lꢅ,
W. Guo, W. Zhang, J. Mol. Model. 2010, 16, 279–284.
Received: May 26, 2011
[25] X. Che, C. Sheng, W. Wang, Y. Cao, Y. Xu, H. Ji, G. Dong, Z. Miao, J. Yao,
W. Zhang, Eur. J. Med. Chem. 2009, 44, 4218–4226.
Published online on July 11, 2011
ChemMedChem 2011, 6, 1806 – 1815
ꢁ 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.chemmedchem.org
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