Synthesis of novel fluconazoliums and their evaluation for antibacterial and antifungal activities

Article abstract of DOI:10.1016/j.ejmech.2011.07.010

A series of novel fluconazoliums were synthesized and their bioactive evaluation as potential antibacterial and antifungal agents were described. Some target compounds displayed good and broad-spectrum antimicrobial activities with low MIC values ranging from 0.25 to 64 μg/mL against all the tested strains, including three Gram-positive bacteria (Staphylococcus aureus, MRSA and Bacillus subtilis), three Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Bacillus proteus) as well as two fungi (Candida albicans and Aspergillus fumigatus). Among all tested title compounds, the octyl, dichlorobenzyl, naphthyl and naphthalimino derivatives gave comparable or even better antibacterial and antifungal efficiency in comparison with the reference drugs Fluconazole, Chloromycin and Norfloxacin.

Full text of DOI:10.1016/j.ejmech.2011.07.010

European Journal of Medicinal Chemistry 46 (2011) 4391e4402
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis of novel fluconazoliums and their evaluation for antibacterial
and antifungal activities
,
b
Yi-Yi Zhang a, Jia-Li Mi c, Cheng-He Zhou a , Xiang-Dong Zhou
*
a Laboratory of Bioorganic & Medicinal Chemistry, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, People’s Republic of China
b School of Pharmaceutical Sciences, Third Military Medical University, Chongqing 400038, People’s Republic of China
c People’s Hospital of Suining, Sichuan 629000, People’s Republic of China
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of novel fluconazoliums were synthesized and their bioactive evaluation as potential antibac-
Received 10 March 2011
Received in revised form
9 June 2011
Accepted 3 July 2011
Available online 8 July 2011
terial and antifungal agents were described. Some target compounds displayed good and broad-
spectrum antimicrobial activities with low MIC values ranging from 0.25 to 64 mg/mL against all the
tested strains, including three Gram-positive bacteria (Staphylococcus aureus, MRSA and Bacillus subtilis),
three Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Bacillus proteus) as well as
two fungi (Candida albicans and Aspergillus fumigatus). Among all tested title compounds, the octyl,
dichlorobenzyl, naphthyl and naphthalimino derivatives gave comparable or even better antibacterial
and antifungal efficiency in comparison with the reference drugs Fluconazole, Chloromycin and
Norfloxacin.
Keywords:
Triazolium
Triazole
Antibacterial
Antifungal
Fluconazole
Ó 2011 Elsevier Masson SAS. All rights reserved.
1. Introduction
in largely extended antimicrobial spectrum and effectively enhance
microbial inhibition. It is generally considered that triazolium is
Triazole compounds have been specially paid increasing atten-
tion because of their extensively medicinal applications in anti-
microbial especially antifungal therapy, and a large number of
predominant triazole drugs have been successfully developed and
prevalently used in the treatment of various microbial infections for
many years [1]. Although the widely clinical use of triazole anti-
infective agents has ever revolutionized the therapeutics of many
infectious diseases, some of them are still limited by poor water
solubility, narrow antimicrobial spectrum and multidrug-
resistance [2,3]. Thus, there is a real perceived need for the devel-
opment of novel and more effective antimicrobial agents.
Triazoliums, the quaternization products of triazolyl ring, are
becoming highlighted topic for that they have been frequently
found to exhibit a variety of biological activities, such as antitumor
[4], antimalaria [5], anti-inflammatory [6], anti-virus [7], antifungal
[8] and antibacterial [9] ones and so on. Particularly in antimicro-
bial aspect, some of them have displayed large probability as
antibacterial and antifungal agents. A lot of works have revealed
that the transformation of triazolyl ring into triazolium could result
able to form hydrogen bonds, accept protons and further produce
electrostatic interaction with biological target sites, and thereby
these inherently structural features could affect the diffusion and
interaction in bacterial tissues with the enhancement of water
solubility and membrane permeability [10]. For example, the
formation of triazolium by the quaternization of triazolyl ring in
Ravuconazole resulted in more potent and broad antimicrobial
behavior compared with Ravuconazole due to the water-soluble
improvement [11]. Some structurally simple triazolium deriva-
tives also exhibited antimicrobial efficiency. Triazolium 1a with
dodecyl moiety showed better activities against Staphylococcus
aureus. Pseudomonas aeruginosa and Bacillus proteus than the
reference drug Chloromycin, and the introduction of 2,4-
dichlorobenyl and carbazolyl moieties by the replacement of
dodecyl group, which yielded corresponding triazolium compounds
1bec, displayed significant antimicrobial efficacy and broad-
spectrum bioactivity against both bacterial and fungal strains
[12,13]. More importantly, mono-triazoliums 1dee were also both
sensitive to fungi and bacteria to some extent [14,15], and bis-
triazoliums 2a and 2b gave 16- to 256-fold more potent efficacy
against Gram-positive and Gram-negative strains in comparison
with Chloromycin [12], while hydrochloride 2c showed better anti-
Candida albicans property than Fluconazole [16]. These implied that
* Corresponding author. Tel./fax: þ86 23 68254967.
E-mail address: zhouch@swu.edu.cn (C.-H. Zhou).
0223-5234/$ e see front matter Ó 2011 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2011.07.010

4392
Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
R1
N
CH3
Cryptococcus, which showed broader antifungal spectrum than
1a, X = Br, R1 = dodecyl, R2 = 2,4-difluorobenzyl
CH3
N
1b, X = Cl, R1 = R2 = 2,4-dichlorobenzyl
1c, X = Cl, R1 = 2,4-dichlorobenzyl,
N
Cl
Fluconazole [22]. Moreover, much work has also focused on the
optimization of triazolyl moiety by diverse functional groups or
investigation of supramolecular drugs of Fluconazole with
different metal ions [23e28]. Nevertheless, to our best knowledge,
the quaternization of N-4 position of triazole ring in Fluconazole,
which produced the corresponding triazolium, has been seldom
observed.
In view of above observation, as an extension of our studies on the
development of triazole antimicrobial agents [16,29e33], it is
reasonablefor uswith great interest to prepare fluconazoliums which
are the quaternization derivatives of two triazolyl rings in Flucona-
zole, and evaluate their antibacterial and antifungal behavior.
Herein a series of novel fluconazoliums were prepared for the
first time (Scheme 1). The design for target fluconazolium mole-
cules is based on the following considerations:
H3C
Cl
CH3
X
HN
N
1e
R
2 = 9-phenylene-9H-carbazole
N
N
Cl
1d, X = Br, R1 = R2 = 4-methoxy-2-hydroxybenzoyl
R2
1a-d
N
N
H
N
N
N
N
N
N
R
N
NH
3Cl
2c
Cl
2Br
Cl
Cl
N
N
2a, R = (CH2)4
2b, R = p-phenyl
N
H
Cl
Cl
Cl
Fig. 1. Structures of some antimicrobial mono- and bis-triazoliums.
the positive triazolium could be helpful for antimicrobial activities,
which have been served as impetus to encourage more researches
on triazolium derivatives (Fig. 1).
It is well known that Fluconazole with two triazole rings, the
first-line antifungal drug recommended by World Health Organi-
zation (WHO), has established an exceptional therapeutic record
for Candida infections with low toxicity, excellent safety profile and
favorable pharmacokinetic characteristics [17]. However, its
extensive use and narrow antifungal spectrum especially low
activity against non-Candida fungi have greatly affected the thera-
peutic efficacy in clinic [18]. Furthermore, the poor water solubility
which precludes its development for intravenous administration
also limited the administrable mode. This situation stimulated the
further structural modification of Fluconazole and a large number
of researches, including esterification of hydroxyl group, modifi-
cation of 2,4-difluorobenzyl and triazolyl moiety etc., have been
devoted to improve its water solubility and/or antimicrobial
spectrum [19e21]. A successful example was Fosfluconazole, an
ester prodrug marketed in 2003 to treat with Candida albicans and
(1) The introduction of two-triazolium entities in combination
with the fluorine atoms and hydroxyl groups in target molecule
should be favorable for modulating physicochemical proper-
ties, such as affinity and water solubility, thus conductive to
antimicrobial activity as literatures have reported, and be
expected to broaden antibacterial and antifungal spectrum and
be beneficial for increasing biological activities.
(2) A variety of alkyl, phenyl, naphthyl and naphthalimido moieties
were incorporated in order to study the effect on antimicrobial
property of the types of the pendent substituents in
fluconazolium.
(3) Different alkyl derivatives were synthesized to explore the
influence of chain lengths on biological activities.
(4) Halogen-containing benzyl groups were reported to signifi-
cantly affect antimicrobial efficacy, therefore
a series of
OH
N
N 2Br
OH
OH
2Br
N
N
N
N
N
F
N
N
N
F
N
N
F
N
N
(vii)
(iv)
N
N
N
N
n
n
(CH2)n
(CH2)n
CH3
10
6
O
O
N
O
N
O
F
CH3
F
F
a, n = 3; b, n = 7;
c
d
, n = 11; , n = 15
(v)
(vi)
OH
OH
2X
N
N
N
a, n = 1; b, n = 4
2Br
N
N
N
Br
Br
N
N
13
N
N
N
N
F
H
F
O
N
O
O
O
O
X2
(ii)
X2
O
O
F
F
11
X1
X2 = 2-F
X1
12
, X = Br, X1 = F,
b, X = Cl, X1 = Cl, X2 = 2-Cl
c, X = Cl, X1 = Cl, X2 = 3-Cl
, X = Cl,
e, X = Cl, X1 = H, X2 = 3-Cl
f, X = Cl,
1 = Cl, X2 = H
, X = PF6, X1 = Cl, X2 = 2-Cl
h, X = Br,
i, X = I,
a
8
Br
Br
(iii)
X
1 = H, X2 = 2-Cl
OH
d
O
N
Br
O
X
n
(i)
Br
g
a
, n = 1
b, n = 4
Br
X
1 = Cl, X2 = 2-Cl
X1 = Cl, X2 = 2-Cl
X
1 = H, X2 = 3-CH3
O
7
9
j
, X = Br,
k, X = Br, X1 = H, X2 = H
l, X = Br, X1 = NO2, X2 = H
Reagents and conditions: (i) 5% NaOH, TBAI, BrCH2CH2Br, 70 oC;
(ii) ammonia water, reflux; (iii) Br(CH2)nBr, DMF, 50 oC; (iv)
CH3(CH2)nBr, CH3CN, 80 oC; (v) substituted benzyl bromide or
chloride, CH3CN, 80 oC; (vi) compound , CH3CN, 80 C; (vii) compound , CH3CN, 80 oC
o
7
9
Scheme 1. Synthesis of target fluconazoliums 10ꢀ13.

Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
4393
halobenzyl moieties were coupled with Fluconazole via qua-
ternization of triazolyl rings.
(5) Naphthalene and naphthalimide rings, which have been dis-
closed to lead the increase of antimicrobial efficacy [34,35], as
chloride produced 2-chloro-1-(2,4-difluorophenyl)ethanone (3),
and then further N-alkylated with 1,2,4-triazole to yield 1-(2,4-
Difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethanone (4). The epox-
ylation of compound 4 by trimethylsulfoxonium iodide afforded
epoxide 5, and was further treated with 1,2,4-triazole to give the
desired intermediate 6 [36]. Bromoethoxy naphthalene 7 was
efficiently prepared by the reaction of 2-naphthol with 1,2-
dibromide in 5% NaOH at 78 ꢁC, while intermediate naph-
thalimide bromides 9aeb were conveniently synthesized from
commercial 4-bromo-1,8-naphthalic anhydride by ammonification
with aqueous ammonia and then N-alkylation with dibromides in
DMF at 50 ꢁC. Compound 6 in acetonitrile reacted with a series of
excess alkyl or aryl halides respectively at 80 ꢁC to yield the target
bis-triazolium derivatives 10e13 (except compounds 11gei). The
2,4-dichlorobenyl fluconazolium 11b was treated in water at room
two strong hydrophobic blocks were easy to form
pep inter-
actions with biological molecules and might affect the lipid/
water partition coefficient of target triazoliums, and thus
further influence the biological activities. Rationally, naphtha-
lene and naphthalimide rings were introduced and evaluated
for their contribution of large
crobial activities.
p-conjugated system to antimi-
(6) Anions play important roles in biological system. Several
different inorganic anions as counterions with fluconazo-
liums were selected to investigate their effect on biological
activity. Compound 11g with PFꢀ6 was specially considered
due to the strong ability of forming hydrogen bond by F
atom.
(7) In comparison with the two-triazolium fluconazolium, some
corresponding mono-triazoliums were also synthesized in
order to survey the roles of triazolium number in antimicrobial
efficacy.
temperature
by saturated
aqueous
ammonium
hexa-
fluorophosphate to give almost quantitatively the corresponding
hexafluorophosphate 11g. Tetrabutyl ammonium bromide and
tetrabutyl ammonium iodide in acetnitrile were added respectively
dropwise to the solution of 11g to afford bromide 11h and iodide
11i in more than 90% yield. All mono-triazole compounds 14 and 15
were easily prepared in acetonitrile in the presence of potassium
carbonate starting from 1,2,4-triazole and 2,4-difluorobenzyl
bromide or intermediates 7 and 9b, and then further coupled
with the corresponding excess mono-bromides via quaternization
of triazolyl ring to produce the desired mono-triazoliums 16e19 in
good yields (Scheme 2).
All the structures of target fluconazoliums including alkyl chain
10, phenyl one 11, naphthyl-containing compound 12 and
naphthalimido-based derivative 13 as well as their corresponding
mono-triazoliums 16e19 were shown in Schemes 1 and 2.
2. Results and discussion
2.2. Analysis of spectra
2.1. Chemistry
All the new compounds were characterized by MS, IR, NMR and
HRMS spectra. Their spectral analyses were consistent with the
assigned structures and listed in the experimental section. The
mass spectra for triazolium derivatives gave a major fragment of
[MeX]þ or [Me2X]þ according to their molecular formula.
The preparation of title fluconazoliums was outlined in Scheme
1. The intermediate Fluconazole 6 was obtained in the yield of 19.2%
via four-step reactions from commercial 2,4-difluorobenzene. The
Friedel-Crafts acylation of 2,4-difluorobenzene with chloroethyl
F
(ii)
N
Br
16
N
N
N
CH3
N
7
Br
N
F
F
F
N
Cl
F
F
F
(i)
(iii)
(iv)
N
F
Br
17
N
14
Cl
F
F
N
Br
N
18
N
O
N
N
Br
4
N
4
4
N
Br
O
N
O
N
F
N
O
N
O
O
N
O
(v)
(vi)
F
19
9b
15
Br
Br
Br
Reagents and conditions: (i) 1,2,4-triazole, K2CO3, CH3CN, 40 oC; (ii) CH3(CH2)7Br, CH3CN,
80 oC; (iii) 2,4-dichlorobenzyl bromide, CH3CN, 80 oC; (iv) compound 7, CH3CN, 80 oC;
(v) 1,2,4-triazole, K2CO3, CH3CN, 60 oC; (vi) 2,4-difluorobenzyl bromide, CH3CN, 80 oC.
Scheme 2. Synthesis of mono-triazoliums 16e19.

4394
Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
2.2.1. IR spectra
shifts (2.01e1.75 ppm for Ha and 1.52e1.32 ppm for Hb) compared
to that of Fluconazole ( 8.31, 7.79 ppm), respectively. In contrast
with alkyl ones 10aed, aryl fluconazoliums 11ael also gave larger
downfield chemical shifts at
5.71e4.97 ppm for Hf protons, while
all the Hg methylene protons at N-1 position of triazolium in flu-
conazoliums exhibited a singlet at 5.11e4.25 ppm in DMSO-d6.
In IR spectra, all the target fluconazoliums 10e13 gave a broad
absorption in 3385ꢀ3355 cmꢀ1 and sharp peak in 1162ꢀ1145 cmꢀ1
which indicated the presence of tertiary alcohol. The aryl charac-
teristic CꢀH bands of mono- and bis-triazoliums appeared in the
region between 3069 and 3011 cmꢀ1 as well as the vibrations of
aromatic frame showed peaks at 1600ꢀ1400 cmꢀ1, while the
absorption bands ascribed to aliphatic CꢀH stretching vibrations
d
d
d
Meanwhile, the chemical shifts for Hg proton decreased as the
following order: compound 11 > compound 12 > compound
13 > compound 10. This result was different from that of Hf and
revealed that the positive charge did not distribute on average in
triazolyl ring. No large differences were found in Hc, Hd and He
chemical shifts for the protons of 2,4-difluorophenyl ring. More-
over, the presence of strong electrophilic nitro group in compound
11l resulted in large downfield chemical shifts of Ha, Hb, Hf and Hg
protons. In addition, all the other aromatic and aliphatic protons
appeared at the appropriate chemical shifts and integral values.
The 1H NMR spectral analyses of mono- and bis-triazoliums also
indicated that the quaternization of triazolyl ring by alkyl or aryl
halides took place on the N-4 position of 1,2,4-triazole, neither N-1
nor N-2 atoms, perhaps because of its smaller steric hindrance and
lower energy which were in accordance with the reaction sites on
N-4 position [37].
were seen at two regions of 2976ꢀ2925 and 2867ꢀ2846 cmꢀ1
.
Furthermore, two sharp and strong peaks at 1527 and 1351 cmꢀ1 in
compound 11l were attributed to the vibration of N]O bond. The
tertiary amide group in naphthalimide derivatives 9, 13, 15 and 19
gave IR signals between 1660 and 1658 cmꢀ1. All the other
absorption bands were also observed at expected regions.
2.2.2. 1H NMR spectra
In 1H NMR spectra, all the protons for the triazolium ring and
methylene groups linked with N-4 position showed remarkable
downfield signals due to the presence of triazolium positive
charges. Some 1H NMR data of fluconazoliums were listed in
Table 1. Protons Ha and Hb of triazolyl ring appeared a singlet at
d
10.32e10.06 and 9.31e9.11 ppm, which displayed large downfield
Table 1
Some NMR data (DMSO-d6,
d
/ppm) of Fluconazole and several title fluconazoliums.
Ha
Hf
OH
N
N
Hg
N
N
N
N
F
R
Hc
R
Hb
Hd
He
2Br
F
Compds
R
1H NMR (
d
/ppm)
13C NMR (
d
/ppm)
Tri 5-C
Ha
Hb
Hc
Hd
He
Hf
Hg
Tri 3-C
151.3
Tri N4-C
6
e
8.31
7.79
7.21
7.14
6.87
4.63
145.6
e
10b
10.20
10.27
9.18
9.18
7.36
7.32
7.14
6.91
6.92
5.00
5.63
4.25
5.00
148.3
144.3
144.2
56.9
56.9
H3C
Cl
6
11h
11j
7.11
7.06
134.9
137.9
Cl
10.28
9.29
7.24
6.82
5.59
5.01
143.9
56.8
H3C
O2N
11k
11l
10.21
10.32
9.20
9.31
7.21
7.44
7.11
7.16
6.92
6.98
5.38
5.70
4.96
5.11
144.2
144.8
143.8
141.2
57.0
57.7
O
12
10.23
10.06
9.21
9.11
7.39
7.38
7.18
7.19
7.01
6.96
5.03
4.97
4.79
4.34
144.2
154.9
143.6
144.2
56.4
56.8
O
N
O
Br
13a

Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
4395
2.2.3. 13C NMR spectra
(Staphylococcus aureus ATCC 6538, Methicillin-resistant S. aureus
N315 (MRSA) and Bacillus subtilis ATCC 21216), three Gram-
negative bacteria (Escherichia coli ATCC 8099, P. aeruginosa ATCC
27853 and B. proteus ATCC 13315) and two fungi (C. albicans ATCC
76615 and Aspergillus fumigatus ATCC 96918) using two folds serial
dilution technique recommended by National Committee for Clin-
ical Laboratory Standards (NCCLS) with the positive control of
clinically antimicrobial drugs Fluconazole, Chloromycin and Nor-
floxacin [38]. The values of clogP, a partition coefficient as a kind of
measure for hydrophobicity/lipophilicity, were calculated using
ChemDraw Ultra 10.0 software integrated with Cambridge soft
Software (Cambridge Soft Corporation). The antibacterial and
antifungal data as well as clogP values were depicted in Table 2.
The 13C NMR spectral analyses were consistent with the assigned
structures. No large differences were found in 13C chemical shifts
for the 2,4-difluorobenzyl group and methylene carbons
(d
48.4e47.1 ppm) which linked with N-1 position of 1,2,4-triazole
ring in fluconazoliums in comparison with that of Fluconazole. The
split signals observed at 164.9e161.1 and 160.7e157.3 ppm were
d
assigned to the C-2 and C-4 position of 2,4-difluorobenzyl group
respectively due to the existence of F atom with strong electroneg-
ativity. The tertiary alcohol carbon resonated at
which ascribed to its proximate oxygen atom with strong electron-
withdrawing property.
The 3-position carbon of triazolium ring in fluconazoliums gave
the downfield 13C chemical shifts with respect to the 5-position,
which clearly suggested that the positive charge should not be
averagely distributed in triazolium ring. Compared with Flucona-
zole, the formation of positive charge triazolium in fluconazoliums
resulted in upfield 13C chemical shifts with 0.6e7.4 ppm for the C-3
position and 0.4e12.1 ppm for the C-5 position of triazolium ring,
whose value size depended on the pended substituents in fluco-
nazoliums, except for compounds 13a and 13b with slight down-
field for the 3-position carbon. Furthermore, the methylene carbon
d 73.1e72.4 ppm,
2.3.1. Antibacterial activity
The antibacterial tests indicated that some title compounds
could effectively, to some extent, inhibit the growth of the tested
microorganisms in vitro, especially toward S. aureus and E. coli with
MIC values ranging from 0.25 to 64
mg/mL. Specially, fluconazo-
liums 10bec, 11bec, 11g, 12 and 13aeb exhibited good to excellent
bioactivities against all the bacterial strains. The results clearly
exhibited that Gram-negative bacteria B. subtilis was less sensitive
to most mono- and bis-triazoliums than other strains, and all the
intermediates including bromides 7 and 9aeb as well as mono-
triazoles 14e15 displayed no obvious activities in inhibiting the
growth of microorganisms.
Results in Table 2 demonstrated the significant effects of alkyl
chain lengths on biological activities. Long-chain alkyl fluconazo-
liums 10bed possessed the better antimicrobial activities to most
tested strains, especially for S. aureus and E. coli with MIC values of
(d
57.1e56.9 ppm) linked with N-4 position of triazolium ring
exhibited larger downfield shifts than that of N-1 position (
d
48.4e47.1 ppm) due to the incorporation of positive charge with
inequable distribution in triazole ring. Moreover, there were no
large differences for the carbon chemical shifts for other groups far
away from triazolium ring (as seen in Table 1).
2.3. Biological activity
1e8
equivalent anti-MRSA potency to Chloromycin (MIC ¼ 8
the strongest antibacterial potencyagainst S. aureus (MIC ¼ 1
m
g/mL. Notably, compound 10b with octyl pendant displayed
g/mL) and
g/mL),
m
The in vitro antimicrobial screening for all synthesized fluco-
nazoliums was evaluated for three Gram-positive bacteria
m
Table 2
Antibacterial and antifungal activities in vitro as MIC (
m
g/mL) for compounds 7 and 9ꢀ19.
Compds
ClogP
Gram-positive bacteria Gram-negative bacteria
Fungi
S. aureus
MRSA
B. subtilis
E. coli
P. aeruginosa
B. proteus
C. albicans
A. fumigatus
7
9a
9b
10a
10b
10c
10d
11a
11b
11c
11d
11e
11f
11g
11h
11i
11j
11k
11l
12
3.97
4.67
6.11
256
128
256
>256
1
2
8
64
8
8
>256
>256
>256
>256
8
>256
>256
>256
>256
32
>256
128
256
16
32
256
64
256
32
32
256
256
>256
128
1
2
8
64
4
32
128
32
256
ꢂ0.25
16
>256
>256
>256
256
8
>256
>256
>256
256
64
>256
>256
>256
128
2
2
4
128
16
4
128
4
256
128
64
64
64
256
128
8
>256
128
256
>256
128
>256
>256
>256
128
128
32
256
64
256
128
32
ꢀ5.07
ꢀ0.83
1.18
32
8
8
NCa
128
256
4
64
256
32
>256
32
64
128
>256
4
16
>256
64
16
64
64
32
256
128
0.5
8
ꢀ6.15
ꢀ3.87
ꢀ4.11
ꢀ5.30
ꢀ5.30
ꢀ5.30
ꢀ3.87
ꢀ3.87
ꢀ3.87
ꢀ5.73
ꢀ6.72
ꢀ7.24
ꢀ0.70
0.30
32
4
8
128
256
1
4
4
256
128
128
4
64
32
64
16
16
256
64
4
64
64
4
256
128
256
32
256
256
128
64
128
256
64
2
8
256
128
128
8
256
128
128
128
16
13a
8
4
64
16
32
8
13b
14
15
NC
1.31
4.29
1.11
8
16
64
8
32
64
16
32
>256
>256
8
>256
>256
16
>256
>256
64
>256
>256
16
>256
>256
16
>256
>256
8
>256
>256
32
>256
>256
32
16
17
18
19
ꢀ0.41
>256
16
16
e
>256
64
64
128
64
128
e
128
8
32
128
8
8
64
16
16
e
>256
32
64
0.5
e
>256
128
64
256
e
1.18
1.43
Fluconazole
Norfloxacin
Chloromycin
ꢀ0.44
e
e
e
0.58
8
64
1
8
1
0.5
64
2
1
16
ꢀ1.09
>256
>256
e
e
a
NC ¼ not calculable.

4396
Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
which was 8- and 64-fold more active than Norfloxacin and Chlor-
omycin, respectively. When the pendant was extended to dodecyl
group, triazolium 10c gave more efficient inhibition against
P. aeruginosa and B. proteus than the standard drug Chloromycin both
much weaker antibacterial property than its corresponding
compound 11b. This seems to reveal that the bis-triazoliums might
be more helpful for bioactivity than mono-triazoliums.
In general, the target fluconazoliums 10bec, 11bec, 11g, 12 and
13aeb showed good or even excellent antibacterial activities in
comparison with the reference drugs against tested bacteria strains.
More importantly, the title compounds have broadened the
potential applications of Fluconazole into antibacterial field.
Meanwhile, the above discussion demonstrated that the antibac-
terial efficacy might be related to hydrophilicity, electropositive
triazolium and various pendants with electron-donor group to
some extent. Further study is necessary to elucidate their mecha-
nism and structure-activity relationships.
at the concentration of 8 mg/mL. Regretfully, butyl-containing fluco-
nazolium 10a was unfavorable for the antibacterial efficiency even at
high concentration. This might be explained by the presence of its
short alkyl chain with low hydrophobicity, making it difficult to
penetrate the cytomembrane and therefore confined the antibacte-
rial activity.
For tested fluconazoliums 11ael bearing a variety of phenyl
derivatives, some compounds gave comparable or superior
inhibitory potency toward the reference drugs Chloromycin and
Norfloxacin, though most were relatively weak to the alkyl ones.
Dichlorobenzyl compounds 11bec displayed better efficacy
against all the tested strains than mono-substituted ones 11def at
2.3.2. Antifungal activity
The antifungal evaluation in vitro exhibited that the activities for
all fluconazoliums were relatively weak compared to their anti-
bacterial activities. Unexpectedly, all target compounds exerted
negative efficacy toward C. albicans when introducing electropos-
itive triazolium. This may be related to the occupation of the
unshared electron pair of 4-N atom in triazole ring, so that the 4-N
atom could not coordinate with Fe2þ ion and thereby result in
unsatisfactory antifungal activity [40]. Despite of this, title
compounds 11d, 11i and 13aeb showed desirable antifungal
activities against A. fumigatus with MIC values between 16 and
the concentrations of 0.5e32 mg/mL, whereas analog 11a showed
weak activities against both Gram-negative and Gram-positive
bacteria. Notably, 2,4-dichlorobenzyl-pended fluconazolium 11b
possessed better efficiency in inhibiting the growth of tested
bacteria than the reference drug Chloromycin to some extent. It
gave low inhibitory concentration (MIC ¼ 4
which was potential as anti-MRSA agent in comparison with
clinical drugs Chloromycin (MIC ¼ 8 g/mL) and Norfloxacin
(MIC ¼ 1 g/mL). Moreover, compound 11b, with prominent MIC
value of 0.5 g/mL against B. proteus, was also confirmed to be the
mg/mL) against MRSA,
m
m
m
32 mg/mL, superior to the first-line antifungal drug Fluconazole.
most active one, 32-fold more potent than Chloromycin. This
manifested that o-substituted halobenzyl derivatives (11b and
11d) might be suitable for antibacterial activity than m-substituted
ones (11c and 11e). Compared with the anion chloride 11b, hex-
afluorophosphate 11g, bromide 11h and iodide 11i were procured
to investigate the effect of different anions on biological activity.
Especially for E. coli, compound 11g gave quite low inhibitory
Otherwise, the antifungal efficiency of mono-triazoliums 16e19
was also tested with less sensitive ability than that of corre-
sponding fluconazolium derivatives, which confirmed the deduc-
tion concluded from the antibacterial test.
2.3.3. Effect of clogP values on antimicrobial activity
These antimicrobial results could also be analyzed by clogP
values. The calculated values of logP for title compounds demon-
strated that compounds with lower absolute values of clogP
showed better antimicrobial activities, and fluconazoliums pos-
sessing comparable clogP values with respect to the reference
drugs exhibited equivalent potency. As seen from Table 2, all flu-
conazoliums showed low clogP values ranging from ꢀ7.24 to 1.18,
meaning that fluconazoliums exhibit good water solubility.
However, lipophilicify was also a very important factor for drug
candidates attributed to its effect on the interaction between
molecule and membrane. The clogP values of the aryl fluconazo-
liums 11ael were at least three orders of magnitude less than that
of the alkyl-pended fluconazoliums 10a (clogP ¼ ꢀ0.83), 12
(clogP ¼ ꢀ0.70) and 13a (clogP ¼ 0.30), in accordance with the
antimicrobial results discussed above, indicating that clogP values
of compounds played significant role in their antimicrobial
efficiency.
concentrations (MIC ꢂ 0.25
m
g/mL) which was 256-fold more
effective than Chloromycin (MIC ¼ 64
mg/mL) and almost equi-
potent to Norfloxacin. However, no large antibacterial difference
for various counterion anions against these tested bacteria was
observed among the compounds 11gei, while compound 11b
displayed a little better potency. On the other hand, compounds
11jel incorporating 3-methylbenzyl, benzyl or 4-nitrobenzyl
group all gave poor activities against tested strains, implying
that the antibacterial activities had been ascribed to the nature of
the pendants to some extent.
Compared to phenyl fluconazoliums 11ael, large aromatic rings
included naphthalene derivative 12 and naphthalimide compounds
13aeb, showed significantly increased antibacterial properties
against most bacteria. Specially, naphthyl group, a large conjugated
system with strong hydrophobic moiety, was incorporated to give
compound 12 with excellent efficacy and broad spectrum at the
concentrations ranging from 2 to 8 mg/mL, which was 4- to 32-fold
more potent than Chloromycin except for MRSA. Naphthalimide
3. Conclusion
fluconazolium 13aeb also displayed good inhibitory abilities
toward tested strains with MIC values of 8e64
m
g/mL, being slightly
In conclusion, a series of novel fluconazoliums including alkyl,
benzyl, naphthethoxyl and naphthalimino moieties were success-
fully synthesized and characterized by NMR, MS, HRMS and IR
spectra. The in vitro antimicrobial activities revealed that most title
compounds could inhibit the growth of tested strains significantly
and even display equipotent or superior activities in comparison
with that of current clinical drugs Fluconazole, Norfloxacin and
weaker than that of naphthyl-containing one 12. This bioactive
difference might be ascribed to the stronger electron-donor of
compound 12, which was an important factor to affect the bio-
logical activity [39]. Furthermore, compound 13a with (CH2)3 linker
was also observed to exhibit the best anti-MRSA efficacy with MIC
value of 4 mg/mL, which was almost equipotent to Norfloxacin.
In order to investigate the influence of number of triazoliums on
antimicrobial activities, mono-triazoliums 16e19 as a library of
controls to fluconazoliums 10e13 were evaluated for their anti-
bacterial activities. All the mono-triazoliums were comparable or
less sensitive to the tested strains in comparison with their corre-
sponding bis-triazoliums, especially mono-triazolium 17 showed
Chloromycin. Noticeably, fluconazoliums 10b (MIC ¼ 1
mg/mL), 11b
g/mL) gave the most
potent antibacterial efficacy against S. aureus, B. proteus and E. coli,
respectively, among all tested compounds including the three
standard drugs. Particularly, compounds 10b, 11b and 13aeb
should also be worthy to be further investigated as potential
(MIC ¼ 0.5
m
g/mL) and 11g (MIC ꢂ 0.25
m

Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
4397
antibacterial agents against MRSA strain, meanwhile compounds
13aeb displayed 8- or 16-fold superior activities toward Flucona-
zole-insensitive A. fumigatus. These results manifested that the
synthesized fluconazoliums had broadened antimicrobial spectrum
into antibacterial field with prominent efficacy compared to the
precursor Fluconazole, which made it a good starting point for the
extension as antimicrobial agents. Further researches, including the
incorporation of various functional groups (ester, ketone, amino
ones etc.), the in vivo bioactive evaluation along with toxicity
investigation as well as the potential prodrug detection are
underway in our group and the findings are expected to report in
due course.
extracted with chloroform. Subsequently, the organic extracts were
collected, dried over anhydrous sodium sulfate and concentrated
under reduced pressure to give the crude material, which was
purified by silica gel column chromatography (eluent, chloroform/
petroleum, 1/8, V/V) to afford the desired naphthalimide bromides
9aeb.
4.1.2.2.1. 2-(4-Bromopropyl)-4-bromo-1,8-naphthalimide
(9a). Compound 9a (1.57 g) was obtained as white solid. Yield:
76.3%; mp: 154e155 ꢁC; IR (KBr)
n
: 3120, 3061 (AreH), 2934, 2866
(CH2), 1658 (C]O), 1588, 1507 (aromatic frame), 1346, 1140, 1051,
864, 754, 750, 610 cmꢀ1; 1H NMR (300 MHz, CDCl3)
: 8.67 (d, 1H,
d
J ¼ 7.1 Hz, NAPH-H), 8.59 (d, 1H, J ¼ 8.3 Hz, NAPH-H), 8.42 (d, 1H,
J ¼ 7.8 Hz, NAPH-H), 8.06 (d, 1H, J ¼ 7.8 Hz, NAPH-H), 7.87 (t, 1H,
J ¼ 7.9 Hz, NAPH-H), 4.24e4.20 (m, 2H, CH2Br), 3.50e3.46 (m, 2H,
Naphthalimide N-CH2), 1.99e1.91 (m, 2H, CH2CH2Br) ppm; ESI-MS
(m/z): 397 [M]þ; HRMS (TOF) calcd. for C15H11Br2NO2 [M þ H]þ,
395.9235; found, 395.9230.
4. Experimental
4.1. General methods
Melting points were uncorrected and were recorded on Xe6
melting point apparatus. TLC analysis was done using pre-coated
silica gel plates. FT-IR spectra were carried out on Bruker RFS100/
S spectrophotometer (Bio-Rad, Cambridge, MA, USA) using KBr
pellets in the 400e4000 cmꢀ1 range. NMR spectra were recorded
on a Bruker AV 300 or Varian 400 spectrometer using TMS as an
4.1.2.2.2. 2-(6-Bromohexyl)-4-bromo-1,8-naphthalimide
(9b). Compound 9b (1.54 g) was obtained as white solid. Yield:
70.0%; mp: 119e120 ꢁC; IR (KBr)
n
: 3107, 3069 (AreH), 2965, 2858
(CH2), 1659 (C]O), 1591, 1570, 1503, 1461 (aromatic frame), 1349,
1231, 1104, 1081, 779, 618 cmꢀ1; 1H NMR (300 MHz, CDCl3)
: 8.66
d
(d, 1H, J ¼ 7.1 Hz, NAPH-H), 8.57 (d, 1H, J ¼ 8.4 Hz, NAPH-H), 8.42 (d,
1H, J ¼ 7.7 Hz, NAPH-H), 8.05 (d, 1H, J ¼ 7.8 Hz, NAPH-H), 7.87 (t, 1H,
J ¼ 7.9 Hz, NAPH-H), 4.20e4.15 (m, 2H, CH2Br), 3.43e3.39 (m, 2H,
Naphthalimide N-CH2), 1.91e1.86 (m, 2H, CH2CH2Br), 1.77e1.73 (m,
2H, Naphthalimide N-CH2CH2), 1.61e1.50 (m, 4H, CH2CH2(CH2)2Br)
ppm; ESI-MS (m/z): 439 [M]þ; HRMS (TOF) calcd. for C18H17Br2NO2
[M þ H]þ, 437.9704; found, 437.9706.
internal standard; Ph
¼
phenyl ring, Tri
¼
triazolyl ring,
NAPH ¼ naphthyl ring, Flu ¼ Fluconazole. The chemical shifts were
reported in parts per million (ppm), the coupling constants (J) are
expressed in hertz (Hz) and signals were described as singlet (s),
doublet (d), triplet (t), quartet (q) as well as multiplet (m). The mass
spectra were recorded on LCMSe2010A and HRMS. All chemicals
and solvents were commercially available and were used without
further purification.
4.1.3. Synthesis of fluconazoliums (10ꢀ13)
4.1.3.1. General procedure for the preparation of alkyl fluconazoliums
(10aꢀd). To a solution of compound 6 (0.12 g, 0.4 mmol) in
acetonitrile (10 mL) was added 1-butyl bromide (0.14 g, 1.0 mmol),
1-octyl bromide (0.19 g, 1.0 mmol), 1-dodecyl bromide (0.25 g,
1.0 mmol), or 1-hexadecyl bromide (0.30 g, 1.0 mmol) respectively.
The mixture was stirred at 80 ꢁC. After the reaction came to the end
(monitored by TLC, eluent, methanol/aqueous ammonia, 10/1, V/V),
the solvent was evaporated under reduced pressure. The residue
was washed three times with petroleum ether and dried in vacuum
to give alkyl fluconazoliums 10aꢀd.
4.1.1. Synthesis of intermediate 2-(2-bromoethoxy)naphthalene (7)
To a solution of 2-naphthol (1.45 g, 10.0 mmol) in 5% NaOH
(10 mL) were added TBAI (0.1 g, 0.27 mmol) and 1,2-dibromoethane
(5.51 g, 30 mmol) successively. The mixture was stirred at 78 ꢁC for
8 h. After the reaction came to the end (monitored by TLC, eluent,
chloroform/petroleum, 1/5, V/V), the mixture was cooled to room
temperature and the solvent was removed. The residue was
extracted with chloroform and the organic extracts were collected,
dried over anhydrous sodium sulfate and purified by silica gel
column chromatography (eluent, petroleum) to afford bromide 7
(1.50 g) as white solid. Yield: 60.2%; mp: 98e99 ꢁC (literature mp:
96e97 ꢁC) [41].
4.1.3.1.1. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-butyl-1H-1,2,4-triazol-4-ium) bromide (10a). The butyl fluco-
nazolium 10a (0.15 g) was obtained as colorless syrup. Yield: 63.8%;
IR (KBr)
n: 3358 (OH), 3110 (AreH), 2925, 2867 (CH2), 1574, 1504,
4.1.2. Synthesis of intermediate naphthalimide bromides (9aeb)
4.1.2.1. 4-Bromo-1,8-naphthalimide (8). A mixture of 4-bromo-1,8-
naphthalic anhydride (2.77 g, 0.01 mol) and aqueous ammonia
(30 mL) was refluxed for 8 h. After the reaction came to the end, the
residue was filtrated. The filter cake was dried in vacuum to give
crude product 4-bromo-1,8-naphthalimide (8, 2.62 g) as brown
solid which was used in following reaction without further purifi-
cation. Yield: 95%; mp: 296e298 ꢁC, in agreement with the
commercial material (mp: 297e298 ꢁC).
1420 (aromatic frame),1383 (CH3),1274,1150 (CeO),1095, 967, 852,
737, 669 cmꢀ1 1H NMR (300 MHz, D2O)
; d: 8.81 (s, 4H, Tri 3,5-H),
7.45e6.61 (m, 3H, 2,4-F2Ph-H), 5.32e5.14 (m, 4H, Tri N1-CH2),
4.32e4.28 (m, 4H, Tri N4-CH2), 1.88e1.70 (m, 4H, Tri N4-CH2CH2),
1.35e1.28 (m, 4H, CH3CH2), 0.86 (t, 6H, J ¼ 6.5 Hz, CH3) ppm; 13C
NMR (100 MHz, DMSO-d6): d 163.5, 161.1 (2,4-F2Ph 4-C), 160.1, 157.7
(2,4-F2Ph 2-C), 150.7 (Tri 3-C), 145.2 (Tri 5-C), 129.7 (2,4-F2Ph 6-C),
123.3 (2,4-F2Ph 1-C), 111.2 (2,4-F2Ph 5-C), 104.1 (2,4-F2Ph 3-C), 73.1
(C-OH), 57.1 (Tri N4-CH2), 47.1 (Tri N1-CH2), 30.8 (Tri N4-CH2CH2),
18.5 (CH2CH3), 13.3 (CH3) ppm; ESI-MS (m/z): 420 [Me2Br]þ;
HRMS (TOF) calcd. for C21H30F2N6O2þ [M þ H]þ, 421.2516; found,
421.2510.
4.1.2.2. General procedure for the preparation of intermediates
naphthalimide bromides (9aeb). To a solution of compound 8
(0.28 g, 1.0 mmol) in DMF (20 mL) was added potassium carbonate
(0.21 g, 1.5 mmol). After the reaction was stirred at 40 ꢁC for an
hour, 1,3-dibromopropane (0.24 g, 1.2 mmol) or 1,6-
dibromopropane (0.29 g, 1.2 mmol) was added, and the resulting
mixture was stirred for another 8 h at 50 ꢁC. After the reaction came
to the end (monitored by TLC, eluent, chloroform/petroleum, 1/4,
V/V), the mixture was cooled to room temperature and the solvent
was removed under reduced pressure, then the residue was
4.1.3.1.2. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-octyl-1H-1,2,4-triazol-4-ium) bromide (10b). The octyl fluco-
nazolium 10b (0.22 g) was obtained as brown syrup. Yield: 78.5%;
IR (KBr)
(aromatic frame), 1383 (CH3), 1274, 1156 (CeO), 1105, 967, 851, 731,
660 cmꢀ1 1H NMR (300 MHz, DMSO-d6)
: 10.20 (s, 2H, Tri 3-H),
n: 3356 (OH), 3117 (AreH), 2927, 2854 (CH2), 1575, 1503
;
d
9.18 (s, 2H, Tri 5-H), 7.40e7.33 (m, 1H, 2,4-F2Ph 6-H), 7.17e7.10 (m,
1H, 2,4-F2Ph 5-H), 6.94e6.89 (m, 1H, 2,4-F2Ph 3-H), 5.00 (s, 4H, Tri

4398
Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
N1-CH2), 4.28e4.23 (m, 4H, Tri N4-CH2), 1.78e1.70 (m, 4H, Tri N4-
CH2CH2), 1.22e1.14 (m, 20H, Tri N4-CH2CH2(CH2)5), 0.84 (t, 6H,
J ¼ 5.7 Hz, CH3) ppm; 13C NMR (100 MHz, DMSO-d6): 163.6, 161.2
(2,4-F2Ph 4-C), 160.2, 157.6 (2,4-F2Ph 2-C), 148.3 (Tri 3-C), 144.2 (Tri
5-C), 129.8 (2,4-F2Ph 6-C), 121.3 (2,4-F2Ph 1-C), 111.3 (2,4-F2Ph 5-C),
104.3 (2,4-F2Ph 3-C), 73.0 (C-OH), 56.9 (Tri N4-CH2), 47.3 (Tri N1-
CH2), 31.7 (Tri N4-CH2CH2), 29.2 (CH2CH2CH2CH2CH2CH3), 29.0
(CH2CH2CH2CH3), 27.5 (CH2CH2CH3), 23.2 (CH2CH3), 14.0 (CH3)
ppm; ESI-MS (m/z): 611 [MꢀBr]þ, 531 [Mꢀ2Br]þ; HRMS (TOF)
calcd. for C29H46Br2F2N6O2þ [M þ H]þ, 533.3768; found, 533.3772.
4.1.3.1.3. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-dodecyl-1H-1,2,4-triazol-4-ium) bromide (10c). The dodecyl
fluconazolium 10c (0.26 g) was obtained as brown oil. Yield: 80.4%;
561 [Mꢀ2Br]þ; HRMS (TOF) calcd. for C27H22F6N6O2þ [M þ H]þ,
561.1827; found, 561.1823.
4.1.3.2.2. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diy
l)bis(4-(2,4-dichlorobenzyl)-1H-1,2,4- triazol-4-ium) chloride (11b).
Prepared according to the general procedure described for 10aꢀd,
starting from compound
6 (0.12 g, 0.4 mmol) and 2,4-
dichlorobenzyl chloride (0.20 g, 1.0 mmol), the pure 2,4-
dichlorobenzyl fluconazolium 11b (0.24 g) was obtained as white
solid. Yield: 86.4%; mp: 120e121 ꢁC; IR (KBr)
n
: 3371 (OH), 3106,
3023 (AreH), 2950, 2852 (CH2), 1507, 1472 (aromatic frame), 1145
(CeO), 768, 620 cmꢀ1; 1H NMR (300 MHz, DMSO-d6)
: 9.92 (s, 2H,
d
Tri 3-H), 8.74 (s, 2H, Tri 5-H), 7.40 (d, 2H, J ¼ 8.0 Hz, 2,4-Cl2Ph 3-H),
7.36 (d, 2H, J ¼ 8.4 Hz, 2,4-Cl2Ph 5-H), 7.28 (d, 2H, J ¼ 8.1 Hz, 2,4-
Cl2Ph 6-H), 6.82e6.75 (m, 1H, 2,4-F2Ph 6-H), 6.67e6.59 (m, 1H,
2,4-F2Ph 5-H), 6.40e6.35 (m, 1H, 2,4-F2Ph 3-H), 5.45e5.30 (m, 4H,
Tri N1-CH2), 5.09e4.81 (m, 4H, Tri N4-CH2) ppm; 13C NMR
IR (KBr)
1462 (aromatic frame), 1383 (CH3), 1274, 1157 (CeO), 1122, 1096,
967, 850, 723, 627 cmꢀ1; 1H NMR (300 MHz, D2O)
: 9.08 (s, 4H, Tri
n: 3355 (OH), 3117 (AreH), 2925, 2855 (CH2), 1575, 1502,
d
3,5-H), 7.16e7.13 (m, 2H, 2,4-F2Ph 5,6-H), 6.93e6.68 (m, 1H, 2,4-
F2Ph 3-H), 5.44e5.24 (m, 4H, Tri N1-CH2), 4.46e4.38 (m, 4H, Tri
N4-CH2), 1.95e1.76 (m, 4H, Tri N4-CH2CH2), 1.22 (s, 36H, Tri N4-
CH2CH2(CH2)9), 0.84 (d, 6H, J ¼ 6.5, CH3) ppm; 13C NMR (100 MHz,
(100 MHz, DMSO-d6): d 163.7, 161.2 (2,4-F2Ph 4-C), 160.2, 157.8 (2,4-
F2Ph 2-C), 144.3 (Tri 3-C), 134.9 (Tri 5-C), 134.1 (2,4-Cl2Ph 4-C), 132.6
(2,4-Cl2Ph 2-C), 130.0 (2,4-Cl2Ph 1-C), 129.3 (2,4-Cl2Ph 3,6-C), 128.0
(2,4-F2Ph 6-C, 2,4-Cl2Ph 5-C), 121.2 (2,4-F2Ph 1-C,), 111.5 (2,4-F2Ph
5-C), 104.3 (2,4-F2Ph 3-C), 72.8 (C-OH), 56.9 (Tri N4-CH2), 48.1 (Tri
N1-CH2) ppm; ESI-MS (m/z): 625 [Mꢀ2Cl]þ; HRMS (TOF) calcd. for
C27H22Cl4F2N6O2þ [M þ H]þ, 625.0645; found, 625.0650.
DMSO-d6): d 163.7, 161.2 (2,4-F2Ph 4-C), 160.2, 157.6 (2,4-F2Ph 2-C),
144.2 (Tri 3-C), 143.7 (Tri 5-C), 129.6 (2,4-F2Ph 6-C), 121.3 (2,4-F2Ph
1-C), 111.2 (2,4-F2Ph 5-C), 104.5 (2,4-F2Ph 3-C), 72.8 (C-OH), 56.9
(Tri N4-CH2), 47.5 (Tri N1-CH2), 35.1 (Tri N4-CH2CH2), 32.2
(CH2(CH2)8CH3), 31.2 (CH2(CH2)7CH3), 29.0 (CH2(CH2)6CH3), 28.1
(CH2CH2CH2(CH2)3CH3), 27.4 (CH2(CH2)2CH3), 25.2 (CH2CH2CH3),
22.0 (CH2CH3), 13.9 (CH3) ppm; ESI-MS (m/z): 645 [Mꢀ2Br]þ;
HRMS (TOF) calcd. for C37H62F2N6O2þ [M þ H]þ, 645.5020; found,
645.5025.
4.1.3.2.3. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl
)bis(4-(3,4-dichlorobenzyl)-1H-1,2,4-triazol-4-ium) chloride (11c).
Prepared according to the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 1-(chloromethyl)-
2-chlorobenzene (0.20 g, 1.0 mmol), pure 3,4-dichlorobenzyl flu-
conazolium 11c (0.23 g) was obtained as syrup. Yield: 82.7%; IR
4.1.3.1.4. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-cetyl-1H-1,2,4-triazol-4-ium) bromide (10d). The cetyl fluco-
(KBr)
(aromatic frame), 1145 (CeO), 1033, 966, 745, 622 cmꢀ1
(300 MHz, D2O)
n
: 3370 (OH), 3106, 3017 (AreH), 2857 (CH2), 1500, 1471, 1402
;
1H NMR
nazolium 10d (0.30 g) was obtained as oil. Yield: 82.7%; IR (KBr)
3356 (OH), 3098 (AreH), 2925, 2854 (CH2), 1503, 1419 (aromatic
frame), 1383 (CH3), 1235, 1151 (CeO), 967, 813, 732, 670 cmꢀ1 1H
NMR (300 MHz, D2O) : 9.04 (s, 4H, Tri 3,5-H), 7.14e6.95 (m, 2H,
n:
d: 8.82 (s, 4H, Tri 3,5-H), 7.57 (d, 2H, J ¼ 7.6 Hz, 3,4-
Cl2Ph 5-H), 7.43 (d, 2H, J ¼ 8.4 Hz, 3,4-Cl2Ph 2-H), 7.18 (d, 2H,
J ¼ 8.3 Hz, 3,4-Cl2Ph 6-H), 6.97e6.90 (m, 1H, 2,4-F2Ph 6-H),
6.84e6.79 (m, 1H, 2,4-F2Ph 5-H), 6.56e6.53 (m, 1H, 2,4-F2Ph 3-H),
5.41 (q, 4H, J ¼ 14.2 Hz, Tri N1-CH2), 5.27e5.02 (m, 4H, Tri N4-CH2)
;
d
2,4-F2Ph 5,6-H), 6.56e6.52 (m, 1H, 2,4-F2Ph 3-H), 5.49e5.21 (m,
4H, Tri N1-CH2), 4.55e4.46 (m, 4H, Tri N4-CH2), 2.03e1.88 (m, 4H,
Tri N4-CH2CH2), 1.78e1.06 (m, 52H, Tri N4-CH2CH2(CH2)13), 0.84 (t,
ppm; 13C NMR (100 MHz, DMSO-d6):
d 163.7, 161.1 (2,4-F2Ph 4-C),
160.2, 157.7 (2,4-F2Ph 2-C), 144.0 (Tri 3-C),134.5 (Tri 5-C),131.8 (3,4-
Cl2Ph 1-C), 131.4 (3,4-Cl2Ph 3-C), 131.0 (3,4-Cl2Ph 2-C), 130.9 (3,4-
Cl2Ph 4-C), 129.1 (3,4-Cl2Ph 6-C), 127.5 (3,4-Cl2Ph 5-C, 2,4-F2Ph 6-
C), 121.2 (2,4-F2Ph 1-C), 111.2 (2,4-F2Ph 5-C), 104.3 (2,4-F2Ph 3-C),
72.7 (C-OH), 57.6 (Tri N4-CH2), 49.0 (Tri N1-CH2) ppm; ESI-MS (m/
z): 625 [Mꢀ2Cl]þ; HRMS (TOF) calcd. for C27H22Cl4F2N6O2þ
[M þ H]þ, 625.0645; found, 625.0649.
6H, J ¼ 6.5 Hz, CH3) ppm; 13C NMR (100 MHz, DMSO-d6):
d 163.6,
161.0 (2,4-F2Ph 4-C), 160.1, 157.7 (2,4-F2Ph 2-C), 148.1 (Tri 3-C),
144.3 (Tri 5-C), 129.4 (2,4-F2Ph 6-C), 121.1 (2,4-F2Ph 1-C), 111.1
(2,4-F2Ph 5-C), 104.7 (2,4-F2Ph 3-C), 72.4 (C-OH), 57.1 (Tri N4-CH2),
48.4 (Tri N1-CH2), 33.5 (Tri N4-CH2CH2), 32.2 (Tri N4-CH2CH2CH2),
29.2 ((CH2)10CH2CH2CH3), 27.4 (CH2CH2CH3), 23.1 (CH2CH3), 13.9
(CH3) ppm; ESI-MS (m/z): 837 [MꢀBr]þ, 757 [Mꢀ2Br]þ; HRMS
4.1.3.2.4. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(2-chlorobenzyl)-1H-1,2,4-triazol- 4-ium) chloride (11d).
Prepared the same way as the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 1-(chloromethyl)-2-
chlorobenzene (0.16 g, 1.0 mmol), pure 2-chlorobenzyl fluconazolium
(TOF) calcd. for C45H78F2N6O2þ [M
757.6265.
þ
H]þ, 757.6272; found,
4.1.3.2. Synthesis of phenyl fluconazoliums (11aꢀj)
4.1.3.2.1. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(2,4-difluorobenzyl)-1H-1,2,4- triazol-4-ium) bromide (11a).
Prepared according to the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 1-(bromo-
methyl)-2,4-difluorobenzene (0.21 g, 1.0 mmol), pure difluor-
obenzyl fluconazolium 11a (0.25 g) was obtained as white solid.
11d (0.21 g) was obtained as syrup. Yield: 82.7%; IR (KBr) n: 3368
(OH), 3113 (AreH), 2950 (CH2), 1501, 1476, 1442 (aromatic frame),
1275, 1150 (CeO), 1056, 967, 748, 623 cmꢀ1; 1H NMR (300 MHz, D2O)
d: 8.83 (s, 4H, Tri 3,5-H), 7.90e7.39 (m, 8H, 2-ClPh-H), 6.89e6.77 (m,
2H, 2,4-F2Ph 5,6-H), 6.65e6.60 (m, 1H, 2,4-F2Ph 3-H), 5.52 (q, 4H,
J ¼ 14.9 Hz, Tri N1-CH2), 5.21e4.97 (m, 4H, Tri N4-CH2) ppm; 13C NMR
Yield: 87.2%; mp: 102e104 ꢁC; IR (KBr)
(AreH), 2846 (CH2), 1508, 1431 (aromatic frame), 1278, 1145
(CeO), 1098, 974, 854, 624 cmꢀ1; 1H NMR (300 MHz, D2O)
: 8.84
n
: 3372 (OH), 3115, 3030
(100 MHz, DMSO-d6): d 163.5, 161.2 (2,4-F2Ph 4-C), 160.2, 158.0 (2,4-
F2Ph 2-C), 144.3 (Tri 3-C), 137.5 (Tri 5-C), 133.7 (2-ClPh 1-C), 132.8
(2-ClPh 2-C), 131.1 (2-ClPh 3-C), 130.6 (2-ClPh 4-C), 129.1 (2-ClPh 6-C),
127.9 (2,4-F2Ph 6-C), 125.4 (2-ClPh 5-C), 121.2 (2,4-F2Ph 1-C) 111.3
(2,4-F2Ph 5-C), 104.4 (2,4-F2Ph 3-C), 72.9 (C-OH), 56.9 (Tri N4-CH2),
49.5 (Tri N1-CH2) ppm; ESI-MS (m/z): 593 [MꢀCl]þ, 557 [Mꢀ2Cl]þ;
HRMS (TOF) calcd. for C27H24Cl2F2N6O2þ [M þ H]þ, 557.1424; found,
557.1428.
d
(s, 4H, Tri 3,5-H), 7.51e6.67 (m, 9H, 2,4-F2Ph-H), 5.49 (q, 4H,
J ¼ 14.7 Hz, Tri N1-CH2), 5.23e4.96 (m, 4H, Tri N4-CH2) ppm; 13C
NMR (100 MHz, DMSO-d6):
d 164.9, 162.5 (2,4-F2Ph 4-C), 160.0,
158.4 (2,4-F2Ph 2-C), 144.9 (Tri 3-C), 133.5 (Tri 5-C), 130.2 (2,4-
F2Ph 6-C), 121.8 (2,4-F2Ph 1-C), 117.8 (2,4-F2Ph 5-C), 112.9 (Flu
2,4-F2Ph 5-C), 112.1 (2,4-F2Ph 3-C), 105.2 (Flu 2,4-F2Ph 3-C), 73.5
(C-OH), 57.8 (Tri N4-CH2), 45.2 (Tri N1-CH2) ppm; ESI-MS (m/z):
4.1.3.2.5. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(3-chlorobenzyl)-1H-1,2,4-triazol- 4-ium) chloride (11e).

Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
4399
Prepared according to the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 1-(chloromethyl)-
3-chlorobenzene (0.16 g, 1.0 mmol) afforded pure 3-chlorobenzyl
stirred at room temperature for 0.5 h. After the reaction came to
the end, the mixture was filtered and washed with acetonitrile for
three times to give pure 2,4-dichlorobenzyl fluconazolium
bromide 11h (0.15 g) as white solid. Yield: 93.3%; mp: 105e107 ꢁC;
fluconazolium 11e (0.20 g) as syrup. Yield: 81.9%; IR (KBr)
(OH), 3111, 3047 (AreH), 2951 (CH2), 1501, 1477, 1430 (aromatic
frame), 1274, 1150 (CeO), 966, 856, 739, 623 cmꢀ1 1H NMR
(300 MHz, D2O) : 8.82 (s, 4H, Tri 3,5-H), 7.46e7.21 (m, 8H, 3-ClPh-
n: 3368
IR (KBr)
1440 (aromatic frame), 1141 (CeO), 967, 861, 735, 623 cmꢀ1
NMR (300 MHz, DMSO-d6) : 10.27 (s, 2H, Tri 3-H), 9.18 (s, 2H, Tri
n
: 3378 (OH), 3110 (AreH), 2927 (CH2), 1573, 1502, 1470,
;
;
1H
d
d
H), 6.97e6.79 (m, 2H, 2,4-F2Ph 5,6-H), 6.65e6.60 (m, 1H, 2,4-F2Ph 3-
5-H), 7.81 (d, 2H, J ¼ 7.9 Hz, 2,4-Cl2Ph 3-H), 7.56 (d, 2H, J ¼ 8.4 Hz,
2,4-Cl2Ph 5-H), 7.48 (d, 2H, J ¼ 8.1 Hz, 2,4-Cl2Ph 6-H), 7.36e7.29
(m, 1H, 2,4-F2Ph 6-H), 7.15e7.07 (m, 1H, 2,4-F2Ph 5-H), 6.94e6.89
(m, 1H, 2,4-F2Ph 3-H), 5.63 (s, 4H, Tri N1-CH2), 5.08e4.92 (m, 4H,
H), 5.44 (q, 4H, J ¼ 13.6 Hz, Tri N1-CH2), 5.26e5.03 (m, 4H, Tri N4-
CH2) ppm; 13C NMR (100 MHz, DMSO-d6):
d 163.4,161.2 (2,4-F2Ph 4-
C),160.1,157.6 (2,4-F2Ph 2-C),144.1 (Tri 3-C),136.0 (Tri 5-C),133.4 (3-
ClPh 1-C), 130.8 (3-ClPh 3-C), 128.9 (3-ClPh 5-C), 128.4 (3-ClPh 2,6-
C), 127.2 (2,4-F2Ph 6-C, 3-ClPh 4-C), 121.2 (2,4-F2Ph 1-C), 111.2 (2,4-
F2Ph 5-C), 104.3 (2,4-F2Ph 3-C), 72.7 (C-OH), 56.9 (Tri N4-CH2), 49.5
(Tri N1-CH2) ppm; ESI-MS (m/z): 593 [MꢀCl]þ, 557 [Mꢀ2Cl]þ; HRMS
(TOF) calcd. for C27H24Cl2F2N6O2þ [M þ H]þ, 557.1424; found,
557.1426.
Tri N4-CH2) ppm; 13C NMR (100 MHz, DMSO-d6):
d 163.6, 161.1
(2,4-F2Ph 4-C), 160.1, 157.7 (2,4-F2Ph 2-C), 144.1 (Tri 3-C), 137.5 (Tri
5-C), 134.1 (2,4-Cl2Ph 4-C), 132.8 (2,4-Cl2Ph 2-C), 130.4 (2,4-Cl2Ph
1-C), 129.0 (2,4-Cl2Ph 3,6-C), 128.2 (2,4-F2Ph 6-C, 2,4-Cl2Ph 5-C),
120.9 (2,4-F2Ph 1-C,), 111.3 (2,4-F2Ph 5-C), 104.3 (2,4-F2Ph 3-C),
72.7 (C-OH), 56.8 (Tri N4-CH2), 48.1 (Tri N1-CH2) ppm; ESI-MS (m/
z): 625 [Mꢀ2Br]þ; HRMS (TOF) calcd. for C27H22Cl4F2N6O2þ
[M þ H]þ, 625.0645; found, 625.0646.
4.1.3.2.6. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(4-chlorobenzyl)-1H-1,2,4-triazol- 4-ium) chloride (11f).
Prepared according to the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 1-(chlor-
omethyl)-4-chlorobenzene (0.16 g, 1.0 mmol) afforded the pure 4-
chlorobenzyl fluconazolium 11f (0.21 g) as syrup. Yield: 85.2%; IR
4.1.3.2.9. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(2,4-dichlorobenzyl)-1H-1,2,4- triazol-4-ium) iodide (11i). To
a solution of compound 11g (0.18 g, 0.2 mmol) in acetonitrile
(15 mL) was added tetrabutyl ammonium iodide (0.079 g,
0.21 mmol, in 2 mL acetonitrile) dropwise. The mixture was stir-
red at room temperature for 0.5 h. After the reaction came to the
end, the mixture was filtered and washed with acetonitrile for
three times to give pure 2,4-dichlorobenzyl fluconazolium iodide
(KBr)
(aromatic frame), 1376, 1151 (CeO), 1092, 962, 851, 743, 618 cmꢀ1
1H NMR (300 MHz, D2O)
: 9.67 (s, 4H, Tri 3,5-H), 7.48e7.23 (m,
n: 3363 (OH), 3109, 3057 (AreH), 2953 (CH2), 1497, 1421
;
d
8H, 4-ClPh-H), 6.99e6.91 (m, 2H, 2,4-F2Ph 5,6-H), 6.74e6.69 (m,
1H, 2,4-F2Ph 3-H), 5.39 (q, 4H, J ¼ 11.1 Hz, Tri N1-CH2), 5.21e5.16
11i (0.16 g) as white solid. Yield: 91.8%; mp: 109e111 ꢁC; IR (KBr)
3370 (OH), 3080 (AreH), 2964 (CH2), 1580, 1474, 1441 (aromatic
frame), 1142 (CeO), 1072, 863, 763, 611 cmꢀ1 1H NMR (300 MHz,
DMSO-d6) : 10.45 (s, 2H, Tri 3-H), 9.17 (s, 2H, Tri 5-H), 7.65 (d, 2H,
n:
(m, 4H, Tri N4-CH2) ppm; 13C NMR (100 MHz, DMSO-d6):
d 163.6,
161.3 (2,4-F2Ph 4-C), 160.3, 158.0 (2,4-F2Ph 2-C), 144.1 (Tri 3-C),
137.7 (Tri 5-C), 133.8 (4-ClPh 1-C), 132.6 (4-ClPh 4-C), 131.1 (4-ClPh
2,6-C), 130.6 (4-ClPh 3,5-C), 129.6 (2,4-F2Ph 6-C), 121.3 (2,4-F2Ph
1-C), 111.4 (2,4-F2Ph 5-C), 104.4 (2,4-F2Ph 3-C), 72.8 (C-OH), 56.9
(Tri N4-CH2), 49.5 (Tri N1-CH2) ppm; ESI-MS (m/z): 593 [MꢀCl]þ,
557 [Mꢀ2Cl]þ; HRMS (TOF) calcd. for C27H24Cl2F2N6O2þ [M þ H]þ,
557.1424; found, 557.1430.
;
d
J ¼ 7.2 Hz, 2,4-Cl2Ph 3-H), 7.54 (d, 2H, J ¼ 5.7 Hz, 2,4-Cl2Ph 5-H),
7.40 (d, 2H, J ¼ 6.4 Hz, 2,4-Cl2Ph 6-H), 7.30e7.23 (m, 1H, 2,4-F2Ph
6-H), 7.15e7.08 (m, 1H, 2,4-F2Ph 5-H), 6.98e6.93 (m, 1H, 2,4-F2Ph
3-H), 5.58 (s, 4H, Tri N1-CH2), 5.06e4.90 (m, 4H, Tri N4-CH2) ppm;
13C NMR (100 MHz, DMSO-d6):
d 163.6, 161.1 (2,4-F2Ph 4-C), 160.1,
4.1.3.2.7. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(2,4-dichlorobenzyl)-1H-1,2,4- triazol-4-ium) hexafluoro-phos-
phate (11g). To a solution of compound 11b (0.28 g, 0.4 mmol) in
water (20 mL) was added aqueous NH4PF6 (0.10 g, 0.6 mmol; in
5 mL water) dropwise. The mixture was stirred at room temper-
ature for 0.5 h. After the reaction came to the end, the solid was
filtered and washed with water for three times and then dried to
give the pure 2,4-dichlorobenzyl fluconazolium hexa-
fluorophosphate 11g (0.35 g) as white powder. Yield: 94.4%; mp:
157.8 (2,4-F2Ph 2-C), 144.1 (Tri 3-C), 134.8 (Tri 5-C), 133.7 (2,4-
Cl2Ph 4-C), 132.4 (2,4-Cl2Ph 2-C), 130.1 (2,4-Cl2Ph 1-C), 129.0
(2,4-Cl2Ph 3,6-C), 128.1 (2,4-F2Ph 6-C, 2,4-Cl2Ph 5-C), 121.1 (2,4-
F2Ph 1-C,), 111.2 (2,4-F2Ph 5-C), 104.4 (2,4-F2Ph 3-C), 72.9 (C-
OH), 56.9 (Tri N4-CH2), 48.1 (Tri N1-CH2) ppm; ESI-MS (m/z): 625
[Mꢀ2I]þ; HRMS (TOF) calcd. for C27H22Cl4F2N6O2þ [M þ H]þ,
625.0645; found, 625.0650.
4.1.3.2.10. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(3-methylbenzyl)-1H-1,2,4- triazol-4-ium) bromide (11j).
Prepared according to the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 1-(bromomethyl)-
3-methylbenzene (0.19 g, 1.0 mmol) afforded the pure 3-
methylbenzyl fluconazolium 11h (0.22 g) as syrup. Yield: 81.6%; IR
115e116 ꢁC; IR (KBr)
1592, 1476, 1451 (aromatic frame), 1145 (CeO), 1072, 867, 773,
623 cmꢀ1 1H NMR (300 MHz, DMSO-d6)
: 10.37 (s, 2H, Tri 3-H),
n: 3378 (OH), 3095, 3054 (AreH), 2976 (CH2),
;
d
9.11 (s, 2H, Tri 5-H), 7.61 (d, 2H, J ¼ 7.2 Hz, 2,4-Cl2Ph 3-H), 7.49 (d,
2H, J ¼ 5.7 Hz, 2,4-Cl2Ph 5-H), 7.38 (d, 2H, J ¼ 6.4 Hz, 2,4-Cl2Ph 6-
H), 7.36e7.31 (m, 1H, 2,4-F2Ph 6-H), 7.17e7.11 (m, 1H, 2,4-F2Ph 5-
H), 6.98e6.93 (m, 1H, 2,4-F2Ph 3-H), 5.67 (s, 4H, Tri N1-CH2),
5.11e4.96 (m, 4H, Tri N4-CH2) ppm; 13C NMR (100 MHz, DMSO-
(KBr)
frame), 1383 (CH3), 1147 (CeO), 1041, 967, 734, 622 cmꢀ1
(300 MHz, DMSO-d6) : 10.21 (s, 2H, Tri 3-H), 9.20 (s, 2H, Tri 5-H),
n
: 3360 (OH), 3009 (AreH), 2925 (CH2), 1503, 1453 (aromatic
;
1H NMR
d
7.45e7.29 (m, 8H, Ph-H), 7.23e7.18 (m, 1H, 2,4-F2Ph 6-H), 7.12e7.09
(m, 1H, 2,4-F2Ph 5-H), 6.95e6.89 (m, 1H, 2,4-F2Ph 3-H), 5.38 (s, 4H,
Tri N1-CH2), 4.98e4.93 (m, 4H, Tri N4-CH2) ppm; 13C NMR (100 MHz,
d6):
d 163.5, 161.1 (2,4-F2Ph 4-C), 160.1, 157.9 (2,4-F2Ph 2-C), 144.5
(Tri 3-C), 134.9 (Tri 5-C), 134.3 (2,4-Cl2Ph 4-C), 132.9 (2,4-Cl2Ph 2-
C), 130.2 (2,4-Cl2Ph 1-C), 129.3 (2,4-Cl2Ph 3,6-C), 127.8 (2,4-F2Ph 6-
C, 2,4-Cl2Ph 5-C), 121.0 (2,4-F2Ph 1-C,), 111.0 (2,4-F2Ph 5-C), 104.6
(2,4-F2Ph 3-C), 72.8 (C-OH), 56.8 (Tri N4-CH2), 48.2 (Tri N1-CH2)
ppm; ESI-MS (m/z): 625 [Mꢀ2PF6]þ; HRMS (TOF) calcd. for
C27H22Cl4F2N6O2þ [M þ H]þ, 625.0645; found, 625.0641.
DMSO-d6):
d 163.6, 161.1 (2,4-F2Ph 4-C), 160.1, 157.6 (2,4-F2Ph 2-C),
143.9 (Tri 3-C),137.9 (Tri 5-C),130.7 (3-CH3Ph 3-C), 129.5 (3-CH3Ph 1-
C), 129.1 (3-CH3Ph 5-C), 128.7 (3-CH3Ph 2-C), 127.5 (3-CH3Ph 6-C),
126.4 (2,4-F2Ph 6-C, 3-CH3Ph 4-C), 121.2 (2,4-F2Ph 1-C), 111.3 (2,4-
F2Ph 5-C), 104.3 (2,4-F2Ph 3-C), 72.7 (C-OH), 56.8 (Tri N4-CH2), 50.1
(Tri N1-CH2), 20.7 (CH3) ppm; ESI-MS (m/z): 515 [Mꢀ2Br] þ; HRMS
(TOF) calcd. for C29H30F2N6O2þ [M þ H]þ, 517.2516; found, 517.2516.
4.1.3.2.11. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-benzyl-1H-1,2,4-triazol-4-ium) bromide (11k). Prepared start-
ing from compound 6 (0.12 g, 0.4 mmol) and 1-bromomethylbenzene
4.1.3.2.8. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(2,4-dichlorobenzyl)-1H-1,2,4- triazol-4-ium) bromide (11h). To
a solution of compound 11g (0.18 g, 0.2 mmol) in acetonitrile
(15 mL) was added tetrabutyl ammonium bromide (0.067 g,
0.21 mmol, in 4 mL acetonitrile) dropwise. The mixture was

4400
Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
(0.19 g,1.0 mmol), the benzyl fluconazolium 11i (0.19 g) was obtained
following the general procedure described for 10aꢀd as brown syrup.
(0.12 g, 0.4 mmol) and 2-(4-bromopropyl)-4-bromo-1,8-
naphthalimide 9a (0.41 g, 1.0 mmol). Yield: 55.2%; mp:
Yield: 74.7%; IR (KBr)
n
: 3364 (OH), 3062 (AreH), 2848 (CH2), 1619,
238e240 ꢁC; IR (KBr)
n
: 3371 (OH), 3134, 3060 (AreH), 2965
(CH2), 1660 (C]O), 1588, 1572, 1501 (aromatic frame), 1346, 1236,
1152 (CeO), 783, 634 cmꢀ1; 1H NMR (300 MHz, DMSO-d6)
: 10.06
1572, 1500, 1455, 1425 (aromatic frame), 1274, 1150 (CeO), 967, 855,
722, 628 cmꢀ1; 1H NMR (300 MHz, DMSO-d6)
d: 10.28 (s, 2H, Tri 3-H),
d
9.29 (s, 2H, Tri 5-H), 7.49e7.33 (m, 10H, Ph-H), 7.28e7.21 (m, 1H, 2,4-
F2Ph 6-H), 7.09e7.01 (m, 1H, 2,4-F2Ph 5-H), 6.85e6.80 (m, 1H, 2,4-
(s, 2H, Tri 3-H), 9.11 (s, 2H, Tri 5-H), 8.55 (d, 4H, J ¼ 6.9 Hz, NAPH-
H), 8.31 (d, 2H, J ¼ 7.8 Hz, NAPH-H), 8.22 (d, 2H, J ¼ 7.8 Hz, NAPH-
H), 8.00 (t, 2H, J ¼ 8.0 Hz, NAPH-H), 7.41e7.34 (m, 1H, 2,4-F2Ph 6-
H), 7.23e7.15 (m, 1H, 2,4-F2Ph 5-H), 6.99e6.93 (m, 1H, 2,4-F2Ph 3-
H), 5.04e4.91 (m, 4H, Tri N1-CH2), 4.34 (t, 4H, J ¼ 7.5 Hz, Tri N4-
CH2), 4.02 (t, 4H, J ¼ 6.0 Hz, Naphthalimide N-CH2), 2.26e2.16 (m,
4H, Naphthalimide N-CH2CH2) ppm; 13C NMR (100 MHz, DMSO-
F2Ph 3-H), 5.59 (s, 4H, Tri N1-CH2), 5.01 (s, 4H, Tri N4-CH2) ppm; 13
C
NMR (100 MHz, DMSO-d6): d 163.7, 161.3 (2,4-F2Ph 4-C), 160.2, 157.7
(2,4-F2Ph 2-C),144.2 (Tri 3-C),143.8 (Tri 5-C),133.7 (Ph 1-C),129.1 (Ph
2,6-C),128.4(Ph 3,5-C),128.0(2,4-F2Ph 6-C),126.4(Ph 4-C),121.0 (2,4-
F2Ph 1-C),111.6 (2,4-F2Ph 5-C),104.8 (2,4-F2Ph 3-C), 72.9 (C-OH), 57.0
(Tri N4-CH2), 50.4 (Tri N1-CH2) ppm; ESI-MS (m/z): 569 [MꢀBr]þ, 487
[Mꢀ2Br]þ; HRMS (TOF) calcd. for C27H26F2N6O2þ [M þ H]þ,
489.2203; found, 489.2204.
d6):
d 163.8 (C]O), 162.5, 161.2 (2,4-F2Ph 4-C), 160.8, 157.7 (2,4-
F2Ph 2-C), 154.9 (Tri 3-C), 144.2 (Tri 5-C), 143.6, 134.4 (NAPH
3,4,9-C), 129.8 (NAPH 10-C), 129.1, 128.0 (NAPH 6,7-C), 127.3 (NAPH
2-C), 124.3 (2,4-F2Ph 1-C), 121.5 (2,4-F2Ph 1-C), 118.9 (NAPH 1-C),
111.8 (NAPH 8-C), 107.5 (2,4-F2Ph 5-C), 104.8 (2,4-F2Ph 3-C), 73.1
(C-OH), 56.8 (Tri N4-CH2), 46.3 (Tri N1-CH2), 37.7 (Naphthalimide
N-CH2), 28.6 (CH2CH2CH2) ppm; ESI-MS (m/z): 1019 [MꢀBr]þ, 939
[Mꢀ2Br]þ; HRMS (TOF) calcd. for C43H34Br2F2N8O25þ [M þ H]þ,
939.1054; found, 939.1046.
4.1.3.2.12. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(4-nitrobenzyl)-1H-1,2,4-triazol-
4-ium) bromide (11l).
Prepared according to the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 1-(bromo-
methyl)-4-nitrobenzene (0.22 g, 1.0 mmol), the pure nitrobenzyl
fluconazolium 11j (0.21 g) was obtained as white solid. Yield:
71.0%; mp: 221e222 ꢁC; IR (KBr)
n
: 3385 (OH), 3181, 3017 (AreH),
2960 (CH2), 1570, 1527, 1422 (aromatic frame), 1351, 1162 (CeO),
1114, 963, 858, 727, 620 cmꢀ1 1H NMR (300 MHz, DMSO-d6)
4.1.3.4.2. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(6-(4-bromo-1,8-naphthalimide)- hexyl)-1H-1,2,4-triazol-4-ium)
bromide (13b). Prepared according to the general procedure
described for 10aꢀd, compound 13b (0.25 g) was obtained as
white solid starting from compound 6 (0.12 g, 0.4 mmol) and 2-
(6-bromohexyl)-4- bromo-1,8-naphthalimide 9b (0.44 g,
;
d:
10.32 (s, 2H, Tri 3-H), 9.31 (s, 2H, Tri 5-H), 8.41e8.39 (m, 4H, 4-
NO2Ph 3,5-H), 7.70e7.68 (m, 4H, 4-NO2Ph 2,6-H), 7.46e7.41 (m,
1H, 2,4-F2Ph 6-H), 7.19e7.12 (m, 1H, 2,4-F2Ph 5-H), 7.02e6.96 (m,
1H, 2,4-F2Ph 3-H), 5.70 (s, 4H, Tri N1-CH2), 5.09e4.94 (m, 4H, Tri
1.0 mmol). Yield: 53.1%; mp: 230e232 ꢁC; IR (KBr)
n: 3378 (OH),
N4-CH2) ppm; 13C NMR (100 MHz, DMSO-d6):
d
163.6, 161.2 (2,4-
3218, 3030 (AreH), 2935, 2860 (CH2), 1658 (C]O), 1501
(aromatic frame), 1349, 1236, 1147 (CeO), 965, 853, 786,
F2Ph 4-C), 160.1, 157.6 (2,4-F2Ph 2-C), 148.3 (4-NO2Ph 4-C), 144.8
(Tri 3-C), 144.6 (4-NO2Ph 1-C), 141.2 (Tri 5-C), 132.7 (4-NO2Ph 2,6-
C), 130.3 (2,4-F2Ph 6-C), 124.4 (4-NO2Ph 3,5-C), 121.5 (2,4-F2Ph 1-
C), 112.1 (2,4-F2Ph 5-C), 105.1 (2,4-F2Ph 3-C), 73.3 (C-OH), 57.7 (Tri
N4-CH2), 50.1 (Tri N1-CH2) ppm; ESI-MS (m/z): 601
[Mꢀ2Br þ Na]þ; HRMS (TOF) calcd. for C27H24F2N8O25þ [M þ H]þ,
579.1905; found, 579.1902.
621 cmꢀ1 1H NMR (300 MHz, DMSO-d6)
; d: 10.05 (s, 2H, Tri 3-H),
9.06 (s, 2H, Tri 5-H), 8.56e8.52 (m, 4H, NAPH-H), 8.29 (d, 2H,
J ¼ 6.0 Hz, NAPH-H), 8.21 (d, 2H, J ¼ 9.0 Hz, NAPH-H), 7.99 (t, 2H,
J ¼ 7.5 Hz, NAPH-H), 7.32e7.29 (m, 1H, 2,4-F2Ph 6-H), 7.19e7.16
(m, 1H, 2,4-F2Ph 5-H), 6.94e6.92 (m, 1H, 2,4-F2Ph 3-H), 4.73 (s,
4H, Tri N1-CH2), 4.22 (t, 4H, J ¼ 6.9 Hz, Tri N4-CH2), 4.00 (t, 4H,
J ¼ 7.2 Hz, Naphthalimide N-CH2), 1.77e1.73 (m, 4H, Tri N4-
CH2CH2), 1.63e1.59 (m, 4H, Naphthalimide N-CH2CH2), 1.35e1.31
(m, 4H, Tri N4-CH2CH2CH2), 1.22e1.20 (m, 4H, Naphthalimide N-
4.1.3.3. Synthesis of naphthyl fluconazolium (12). The desired 1,10-(2-
(2,4-difluorophenyl)-2-hydroxypropane-1,3-diyl)bis(4-(2-(naphthal-
en-2-yloxy)ethyl) -1H-1,2,4-triazol-4-ium) bromide (12) was
prepared according to the general procedure described for 10aꢀd,
starting from compound 6 (0.12 g, 0.4 mmol) and 2-(2-bromoethoxy)
naphthalene 7 (0.25 g, 1.0 mmol), the pure naphthyl fluconazolium 12
(0.14 g) was obtained as white solid. Yield: 41.8%; mp: 116e118 ꢁC; IR
CH2CH2CH2) ppm; 13C NMR (100 MHz, DMSO-d6):
d 163.7 (C]O),
162.4, 161.1 (2,4-F2Ph 4-C), 160.7, 157.8 (2,4-F2Ph 2-C), 154.5 (Tri
3-C), 144.0 (Tri 5-C), 143.1, 133.9 (NAPH 3,4,9-C), 129.6 (NAPH 10-
C), 128.9, 128.1 (NAPH 6,7-C), 127.2 (NAPH 2-C), 124.5 (2,4-F2Ph 6-
C), 121.4 (2,4-F2Ph 1-C), 118.8 (NAPH 1-C), 111.7 (NAPH 8-C), 107.4
(2,4-F2Ph 5-C), 104.6 (2,4-F2Ph 3-C), 73.2 (C-OH), 56.7 (Tri N4-
CH2), 46.2 (Tri N1-CH2), 37.7 (Naphthalimide N-CH2), 27.2 (Tri N4-
CH2CH2), 26.9 (Naphthalimide N-CH2CH2), 25.4 (Tri N4-
CH2CH2CH2CH2) ppm; ESI-MS (m/z): 1021 [Mꢀ2Br]þ; HRMS
(TOF) calcd. for C49H46Br2F2N8O25þ [M þ H]þ, 1023.1993; found,
1023.1994.
(KBr)
1399 (aromatic frame), 1150 (C-O), 966, 616 cmꢀ1; 1H NMR (300 MHz,
DMSO-d6) : 10.23 (s, 2H, Tri 3-H), 9.21 (s, 2H, Tri 5-H), 7.87e7.78 (m,
n: 3380 (OH), 3210, 3086 (Ar-H), 2942 (CH2), 1600, 1575, 1463,
d
7H, NAPH-H), 7.49e7.27 (m, 8H, NAPH-H, 2,4-F2Ph 6-H), 7.20e7.16 (m,
1H, 2,4-F2Ph 5-H), 7.07e6.96 (m, 1H, 2,4-F2Ph 3-H), 5.03 (s, 4H, Tri N1-
CH2), 4.79 (s, 4H, Tri N4-CH2), 4.48e4.41 (m, 4H, CH2O) ppm; 13C NMR
(100 MHz, DMSO-d6): d 163.1 (2,4-F2Ph 4-C), 160.7, 157.3 (2,4-F2Ph 2-
C), 154.9 (NAPH 2-C), 144.2 (Tri 3-C), 143.6 (Tri 5-C), 133.3 (NAPH
4,9,10-C), 128.9 (2,4-F2Ph 6-C), 127.0 (NAPH 5-C), 123.4 (NAPH 7,8-C),
120.5 (2,4-F2Ph 1-C), 118.0 (NAPH 6-C), 110.5 (NAPH 3-C), 106.6 (2,4-
F2Ph 5-C), 104.1 (2,4-F2Ph 3-C), 72.4 (C-OH), 64.6 (OCH2), 56.4 (Tri
N4-CH2), 46.8 (Tri N1-CH2) ppm; ESI-MS (m/z): 727 [MꢀBr]þ, 647
[Mꢀ2Br]þ; HRMS (TOF) calcd. for C37H34F2N6O32þ [M þ H]þ, 649.2728;
found, 649.2733.
4.1.4. General procedure for the preparation of intermediate mono-
triazoles (14ꢀ15)
To
a stirred suspension of potassium carbonate (1.04 g,
7.5 mmol) in acetonitrile (10 mL) was added 1H-1,2,4-triazole
(0.50 g, 7.0 mmol). The mixture was stirred at 60 ꢁC for 1 h.
When the reaction was cooled to room temperature, 2,4-
difluorobenzyl bromide (1.45 g, 7.0 mmol) or compound 9b
(3.07 g, 7.0 mmol) was added, and the resulting mixture was stirred
at 45 ꢁC. After the reaction came to the end (monitored by TLC,
eluent, chloroform/methanol, 10/1, V/V), the solvent was evapo-
rated and the mixture was partitioned between chloroform and
saturated aqueous sodium bicarbonate, the combined organic
extracts were washed with brine, dried over sodium sulfate and
filtered, the filtrate was concentrated. The crude product was
4.1.3.4. Synthesis of naphthalimide-based fluconazoliums (13aꢀb)
4.1.3.4.1. 1,10-(2-(2,4-Difluorophenyl)-2-hydroxypropane-1,3-diyl)
bis(4-(3-(4-bromo-1,8-naphthalimide)- propyl)-1H-1,2,4-triazol-4-
ium) bromide (13a). Prepared according to the general procedure
described for 10aꢀd, naphthalimide-derived fluconazolium 13a
(0.25 g) was obtained as white solid starting from compound 6

Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
4401
purified by silica gel column chromatography (eluent, chloroform/
methanol, 20/1, V/V) to give mono-triazoles 14ꢀ15 respectively.
367 [MꢀBr]þ; HRMS (TOF) calcd. for C21H18F2N3Oþ [M þ H]þ,
367.1491; found, 367.1494.
4.1.4.1. 1-(2,4-Difluorobenzyl)-1H-1,2,4-triazole (14). Mono-triazole
14 (0.85 g) was obtained as white solid. Yield: 62.0%; mp: 56e58 ꢁC;
4.1.5.4. 1-(6-(4-Bromo-1,8-naphthalimide-2-yl)hexyl)-4-(2,4-difluor
obenzyl)-1H-1,2,4-triazol-4-ium bromide (19). Naphthalimide-deri-
ved mono-triazolium 19 (0.88 g) was obtained as white solid
according to the general procedure described for 10aꢀb starting
from 2,4-difluorobenzylbromide (1.26 g, 5.0 mmol) and compound
1H NMR (400 MHz, CDCl3)
d: 8.13 (s, 1H, Tri 3-H), 7.96 (s, 1H, Tri 5-
H), 7.29e7.27 (m, 1H, 2,4-F2Ph 6-H), 6.93e6.86 (m, 2H, 2,4-F2Ph 3,5-
H), 5.36 (s, 2H, 2,4-F2PhCH2) ppm; ESI-MS (m/z): 195 [M]þ; HRMS
(TOF) calcd. for C9H7F2N3 [M þ H]þ, 196.0686; found, 196.0685.
15 (0.78 g, 4.0 mmol). Yield: 69.0%; mp: 212e213 ꢁC; IR (KBr)
3090, 3011 (AreH), 2946 (CH2), 1661 (C]O), 1509 (aromatic frame),
1381, 1234, 1147, 1095, 1019, 856, 779, 637 (CeBr) cmꢀ1 1H NMR
(400 MHz, DMSO-d6) : 10.17 (s, 1H, Tri 3-H), 9.29 (s, 1H, Tri 5-H),
n:
4.1.4.2. 2-(6-(1H-1,2,4-Triazol-1-yl)hexyl)-4-bromo-1,8-naphthalimide
;
(15). Naphthalimide-derived mono-triazole 15 (2.27 g) was obtained
d
as white solid. Yield: 76.0%; mp: 181e182 ꢁC; IR (KBr)
n
: 3100 (AreH),
2981, 2934 (CH2), 1655 (C]O), 1589, 1507, 1457 (aromatic frame),
1362, 1223, 869, 779, 749, 679 cmꢀ1 1H NMR (300 MHz, CDCl3)
8.58 (d, 2H, J ¼ 6.4 Hz, NAPH-H), 8.35 (d, 1H, J ¼ 10.0 Hz, NAPH-H),
8.25 (d, 1H, J ¼ 6.0 Hz, NAPH-H), 8.02 (t, 1H, J ¼ 8.0 Hz, NAPH-H),
7.68e7.64 (m, 1H, 2,4-F2Ph 6-H), 7.42e7.37 (m, 1H, 2,4-F2Ph 5-H),
7.23e7.20 (m, 1H, 2,4-F2Ph 3-H), 5.54 (s, 2H, Tri N4-CH2), 4.36 (t, 2H,
J ¼ 7.2 Hz, Tri N1-CH2), 4.01 (t, 2H, J ¼ 7.2 Hz, Naphthalimide N-CH2),
1.88e1.85 (m, 2H, Tri-CH2CH2), 1.65e1.62 (m, 2H, Naphthalimide N-
CH2CH2), 1.37e1.31 (m, 4H, Tri-(CH2)2CH2CH2) ppm; ESI-MS (m/z):
555 [MeBr]þ; HRMS (TOF) calcd. for C27H24BrF2N4Oþ2 [M þ H]þ,
554.1123; found, 554.1119.
;
d:
8.65 (d, 1H, J ¼ 6.3 Hz, NAPH-H), 8.57 (d, 1H, J ¼ 7.2 Hz, NAPH-H), 8.41
(d,1H, J ¼ 5.7 Hz, NAPH-H), 8.08 (s,1H, Tri 3-H), 8.04 (d,1H, J ¼ 6.0 Hz,
NAPH-H), 7.93 (s, 1H, Tri 5-H), 7.87e7.83 (m, 1H, NAPH-H), 4.19e4.13
(m, 4H, Tri-CH2(CH2)4CH2), 1.95e1.90 (m, 2H, Tri-CH2CH2), 1.75e1.69
(m, 2H, Naphthalimide N-CH2CH2), 1.48e1.36 (m, 4H, Tri-
(CH2)2CH2CH2) ppm; ESI-MS (m/z): 451 [M þ Na]þ, 427 [M þ H]þ;
HRMS (TOF) calcd. for C20H19BrN4O2 [M þ H]þ, 427.0770; found,
427.0770.
4.2. Antibacterial and antifungal assays
4.1.5. Synthesis of mono-triazoliums (16ꢀ19)
Minimal inhibitory concentration (MIC, mg/mL) is defined as
4.1.5.1. 1-(2,4-Difluorobenzyl)-4-octyl-1H-1,2,4-triazol-4-ium bromide
(16). Mono-triazolium 16 was synthesized according to the
general procedure described for 10aꢀb, starting from the
compound 14 (0.78 g, 4.0 mmol) and 1-octyl bromide (0.97 g,
5.0 mmol), the pure product 16 (1.06 g) was obtained as white
solid. Yield: 68.3%; mp: 219e224 ꢁC; 1H NMR (400 MHz, DMSO-d6)
the lowest concentration of target compounds that completely
inhibited the growth of bacteria, by means of standard two folds
serial dilution method in 96-well microtest plates according to
the NCCLS. The tested microorganism strains were provided by
the School of Pharmaceutical Sciences, Southwest University and
the College of Pharmacy, Third Military Medical University. Flu-
conazole, Chloromycin and Norfloxacin were used as control
drugs. DMSO with inoculation bacterial not medicine was used as
positive control to ensure that the solvent had no effect on
bacteria growth. All the bacteria and fungi growth was moni-
tored visually and spectrophotometrically, and the experiments
d
: 11.63 (s, 1H, Tri 3-H), 9.05 (s, 1H, Tri 5-H), 7.73e7.69 (m, 1H, 2,4-
F2Ph 6-H), 6.90e6.84 (m, 1H, 2,4-F2Ph 5-H), 6.82e6.79 (m, 1H, 2,4-
F2Ph 3-H), 5.80 (s, 2H, 2,4-F2PhCH2), 4.51e4.43 (t, 2H, J ¼ 7.6 Hz,
NCH2CH2), 1.94e1.89 (t, 2H, J ¼ 7.2 Hz, NCH2CH2), 1.27e1.18 (br,
18H, (CH2)9CH3), 0.84e0.81 (t, 3H, J ¼ 6.0 Hz, CH3) ppm; ESI-MS
(m/z): 364 [MeBr]þ; HRMS (TOF) calcd. for C17H24F2N3þ
[M þ H]þ, 309.2011; found, 309.2014.
were performed in triplicate. The MIC values in
summarized in Table 2.
mg/mL were
4.1.5.2. 4-(2,4-Dichlorobenzyl)-1-(2,4-difluorobenzyl)-1H-1,2,4-triazol
-4-ium bromide (17). Compound 17 (1.04 g) was prepared following
the general procedure described for 10aꢀb, starting from 2,4-
dichloro-1-(chloromethyl)benzene (1.0 g, 5.0 mmol) and compound
14 (0.78 g, 4.0 mmol), the pure mono-triazolium 17 was obtained as
white solid. Yield: 60.0%; mp: 176e178 ꢁC; 1H NMR (400 MHz,
4.2.1. Antibacterial assays
The prepared compounds 7 and 9e19 were evaluated for their
antibacterial activities against Gram-positive bacteria (S. aureus
ATCC 6538, Methicillin-resistant S. aureus N315 (MRSA) and B. sub-
tilis ATCC 21216), Gram-negative bacteria (E. coli ATCC 8099, P.
aeruginosa ATCC 27853 and B. proteus ATCC 13315). The bacterial
suspension was adjusted with sterile saline to a concentration of
1 ꢃ 105 CFU. Initially the compounds were dissolved in DMSO to
prepare the stock solutions, then the tested compounds and
reference drugs were prepared in MuellereHinton broth (Guang-
dong huaikai microbial sci. & tech co., Ltd, Guangzhou, Guangdong,
China) to obtain the required concentrations of 256, 128, 64, 32, 16,
DMSO-d6) d: 11.85 (s, 1H, Tri 3-H), 8.70 (s, 1H, Tri 5-H), 8.05 (d, 1H,
J ¼ 8.4 Hz, 2,4-Cl2Ph 6-H), 7.70e7.63 (m, 1H, 2,4-F2Ph 6-H), 7.36 (d,
1H, J ¼ 2.0 Hz, 2,4-Cl2Ph 5-H), 7.30e7.27 (m, 1H, 2,4-F2Ph 5-H),
7.00e6.86 (m, 1H, 2,4-Cl2Ph 3-H), 6.79e6.73 (m, 1H, 2,4-F2Ph 3-H),
5.91 (s, 2H, 2,4-F2PhCH2), 5.72 (s, 2H, 2,4-Cl2PhCH2); ESI-MS (m/z):
354 [MeBr]þ; HRMS (TOF) calcd. for C16H12Cl2F2Nþ3 [M þ H]þ,
355.0449; found, 355.0452.
8, 4, 2, 1, 0.5, 0.25
mg/mL. These dilutions were inoculated and
incubated at 37 ꢁC for 24 h.
4.1.5.3. 1-(2,4-Difluorobenzyl)-4-(2-(naphthalen-2-yloxy)ethyl)-1H-
1,2,4-triazol-4-ium bromide (18). Naphthalene-derived mono-
triazolium 18 (1.45 g) was obtained as brown solid according to
the general procedure described for 10aꢀb starting from
compound 7 (1.26 g, 5.0 mmol) and compound 14 (0.78 g,
4.2.2. Antifungal assays
The newly synthesized compounds 7 and 9e19 were evalu-
ated for their antifungal activities against C. albicans ATCC 76615
and A. fumigatus ATCC 96918. A spore suspension in sterile
distilled water was prepared from 1-day old culture of the fungi
growing on Sabouraud agar (SA) media. The final spore concen-
tration was 1e5 ꢃ 103 spore mLꢀ1. From the stock solutions of the
tested compounds and reference antifungal drug Fluconazole,
dilutions in sterile RPMI 1640 medium (Neuronbc Laboraton
Technology CO., Ltd, Beijing, China) were made resulting in
4.0 mmol). Yield: 81.3%; mp: 218e219 ꢁC; IR (KBr)
(AreH), 2964 (CH2), 1570, 1452, 1400 (aromatic frame), 1112, 978,
770, 616 cmꢀ1; 1H NMR (300 MHz, DMSO-d6)
: 10.40 (s, 1H, Tri 3-
n: 3218, 3090
d
H), 9.33 (s, 1H, Tri 5-H), 7.87e7.79 (m, 3H, NAPH-H), 7.70e7.62 (m,
1H, NAPH-H), 7.49e7.44 (m, 1H, 2,4-F2Ph 6-H), 7.40e7.36 (m, 3H,
NAPH-H), 7.24e7.15 (m, 2H, 2,4-F2Ph 3,5-2H), 5.72 (s, 2H, Tri N4-
CH2), 4.77 (s, 2H, Tri N1-CH2), 4.50 (s, 2H, CH2O) ppm; ESI-MS (m/z):
eleven wanted concentrations (0.25e256 mg/mL) of each tested

4402
Y.-Y. Zhang et al. / European Journal of Medicinal Chemistry 46 (2011) 4391e4402
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Acknowledgments
This work was partially supported by Natural Science Founda-
tion of Chongqing (CSCT: 2007BB5369, 2009BB5296) and South-
west University (SWUB2006018, XSGX0602).
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