Synthesis and Biological Evaluation of Triazole Derivatives as Potential Antifungal Agent

Article abstract of DOI:10.1111/j.1747-0285.2012.01398.x

A series of triazole antifungal agents with piperidine side chains were designed and synthesized. Results of preliminary antifungal tests against eight human pathogenic fungi in vitro showed that all the title compounds exhibited excellent activities with broad spectrum. Moreover, a molecular model for the binding between compound 12 and the active site of CACYP51 was provided based on the computational docking results. The side chain of the compound 12 is oriented into substrate access channel 2 (FG loop) and forms hydrophobic and van der waals interactions with surrounding hydrophobic residues. The phenyl group of the side chain can interact with the phenyl group of Phe380 through the formation of π-π face-to-edge interaction. A series of triazole antifungal agents with piperidine side chains were synthesized. And a molecular model for the binding between compound 12 and the active site of CACYP51 was provided based on the computational docking results.

Full text of DOI:10.1111/j.1747-0285.2012.01398.x

ª 2012 John Wiley & Sons A/S
doi: 10.1111/j.1747-0285.2012.01398.x
Chem Biol Drug Des 2012; 80: 382–387
Research Article
Synthesis and Biological Evaluation of Triazole
Derivatives as Potential Antifungal Agent
Xiaoyun Chai1,†, Guang Yang2,†, Jun
The incidence of systemic fungal infections such as Candidosis,
Cryptococcosis, and Aspergillosis has been increasing in prevalence
recently and is associated with the increase in the number of
immunocompromised hosts. Opportunistic and invasive fungal infec-
Zhang3, Shichong Yu1, Yan Zou1, Qiuye
Wu1, Dazhi Zhang1, Yuanying Jiang4,
Yongbing Cao4,* and Qingyan Sun1,*
tions have become the important causes of morbidity and mortality
1Department of Organic Chemistry, School of Pharmacy, Second
(1,2). In addition, the alarming rates of the growing emergence of
antifungal resistance in hospitals are major concerns to the public
health and scientific communities worldwide (3,4). Therefore, all
these trends have emphasized the urgent need for new, more effec-
tive, and safe antifungal agents.
Military Medical University, Guohe Road 325, Shanghai 200433, China
2Pharmacy Team, Administrative Brigade of Postgraduate, Second
Military Medical University, Zhengtong Road 41, Shanghai 200433,
China
3Overseas Education Faculty of the Second Military Medical
University, Xiangyin Road 800, Shanghai 200433, China
4Drug Research Center, School of Pharmacy, Second Military
Medical University, Guohe Road 325, Shanghai 200433, China
*Corresponding author: Qingyan Sun, sqy_2000@163.com
These authors contributed equally to this work.
Among the attractive approaches to find novel antifungal agents,
the structural modification or optimization of the existing agents
has provoked special interest in the realm of medical chemistry.
One kind of them used widely and efficiently is azoles that are
increasing in number and diversity, such as fluconazole (FCZ), itraco-
nazole (ICZ), ravuconazole (VCZ), and posaconazole. FCZ, the pre-
dominant azole agent, is a water-soluble triazole and also has a
very low incidence of side effects. However, the widespread use of
antifungal drugs and their resistance against fungal infections has
led to serious health problems (5–7). In recent years, some of the
current azole antifungal drugs are designed and used in clinic. But
either the highly toxic or the low bioavailability restrains its usage,
such as ICZ and VCZ. Therefore, the search for a novel, more effec-
tive antifungal agent with lower toxicity continues to be an area of
investigation into medicinal chemistry.
A series of triazole antifungal agents with piperi-
dine side chains were designed and synthesized.
Results of preliminary antifungal tests against
eight human pathogenic fungi in vitro showed that
all the title compounds exhibited excellent activi-
ties with broad spectrum. Moreover, a molecular
model for the binding between compound 12 and
the active site of CACYP51 was provided based on
the computational docking results. The side chain
of the compound 12 is oriented into substrate
access channel 2 (FG loop) and forms hydrophobic
and van der waals interactions with surrounding
hydrophobic residues. The phenyl group of the
side chain can interact with the phenyl group of
Phe380 through the formation of p-p face-to-edge
interaction.
Azoles exert antifungal activity through inhibition of the cytochrome
P450 14a-demethylase (CYP51), which is crucial in the process of
biosynthesis of ergosterol by a mechanism in which the heterocyclic
nitrogen atom (N-4 of 1,2,4-triazole) binds to the heme iron atom (8).
In our previous research, we have designed highly potent azole deriv-
atives with different C-3 side chains (9–14). In the present study, we
choose compound B1 [with the MIC80 value of 0.25 mg ⁄ mL against
Candida albicans (C. alb)] as the lead compound (Figure 1) (15). We
focused on modifying the side chain without loss of key interactions.
As we all know the importance of the piperazinyl for the antifungal
activity (12,14), we sought to investigate this further with heterocy-
cles piperidinol, containing an oxygen atom (Figure 2). This replace-
ment was based on the following considerations: (i) the oxygen atom
could improve the flexibility of the molecule, making the side chain
more easily lock its proper position; and (ii) the oxygen atom may
interact with the amino acid residues through hydrogen bonding. We
designed all the title compounds to keep the preferred combination
of the piperidinol at C-3. Besides, we focused our attention on
installing various substituted acids containing aromatic ring, such as
benzenes, pyridines, and furans.
Key words: antifungal agent, molecular docking, synthesis, triazole
Abbreviations: CYP51, the cytochrome P450 14a-demethylase;
MICs, the minimal inhibitory concentrations; C. alb, Candida albicans;
C. par, Candida parapsilosis; C. tro, Candida tropicalis; Cry. neo, Crypto-
coccus neoformans; F. com, Fonsecaea compacta; T. rub, Trichophyton
rubrum; M. gyp, Microsporum gypseum; A. fum, Aspergillus fumigatus;
FCZ, fluconazole; ICZ, itraconazole; VCZ, voriconazole; Mp, melt point;
1H NMR, hydrogen nuclear magnetic resonance; 13C NMR, carbon
nuclear magnetic resonance; IR, infrared spectrum; API-ES atmospheric
pressure electrospray ionization mass spectrometry; NCCLS, National
Committee for Clinical Laboratory Standards.
Received 9 November 2011, revised 7 April 2012 and accepted for pub-
lication 17 April 2012
382

Triazole as Potential Antifungal Agent
added and refluxed for 6 h. The reaction was monitored by TLC.
After filtration, the filtrate was evaporated under reduced pressure.
Water (30 mL) was added to the residue, which was then extracted
with ethyl acetate (80 mL · 3). The extract was washed with satu-
rated NaCl solution (20 mL · 3), dried over anhydrous Na2SO4, and
evaporated. The residue was separated and purified readily by chro-
matography on silica gel to afford compound 5, 1.37 g.
Figure 1: Structure of the lead compound B1.
General procedure for the preparation of the
compounds 6–37
Methods and Materials
To a stirred mixture of 1-[2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-
1,2,4-triazol-1-yl)-propyl]-piperidin-4-ol (5) (0.338 g, 0.001 mol),
DMAP (100 mg) and EDCI (200 mg) in 50 mL of dichloromethane
under 0 ꢀC and substituted acid (0.001 mol) were added. After 1 h,
the reaction was heated and refluxed for 8 h. The reaction was
monitored by TLC. After filtration, the filtrate was evaporated under
reduced pressure. The residue was then extracted with ethyl ace-
tate (60 mL · 3). The extract was washed with saturated NaCl
solution (20 mL · 3), dried over anhydrous Na2SO4, and evaporated.
The residue was crystallized from ethyl acetate to afford the title
compounds, in 67.0–81.0% yield (Appendix S1).
All reactions were monitored by thin-layer chromatography (Huang-
hai, Yantai, China). Compounds were visualized by UV light (Yukang,
Shanghai, China). Melting points were measured on a YamatoMP-
21 melting-point apparatus and are uncorrected. Infrared spectra
were recorded in potassium bromide disks on a HITACHI270-50
spectrophotometer. NMR spectra were recorded on a Bruker AC-
1
500P spectrometer at 500 MHz for H NMR and 125 MHz for 13C
NMR with TMS as the internal standard. Chemical shifts are
expressed in d (ppm). Column chromatography was carried out
using silica gel (300–400 mesh) (Huanghai, Yantai, China). Silica gel
chromatography solvents were of analytical grade. Atmospheric
pressure electrospray ionization mass spectrometry (API-ES) experi-
ment was carried out in an API-3000 LC-MS spectrometer. All reac-
tion solvents were dried prior to use according to standard
procedures.
Biological activity
The in vitro antifungal activities of all the title compounds were
evaluated against eight human pathogenic fungi, C. alb, Candida
parapsilosis (C. par), Candida tropicalis (C. tro), Cryptococcus neofor-
mans (Cry. neo), Fonsecaea compacta (F. com), Trichophyton rubrum
(T. rub), Microsporum gypseum (M. gyp), and Aspergillus fumigates
(A. fum), which are often encountered clinically, and were compared
with FCZ, ICZ, and VCZ. C. alb and Cry. neo were provided by
Shanghai Changzheng Hospital; C. par, C. tro, F. com, T. rub,
M. gyp, and A. fum were provided by Shanghai Changhai Hospital.
C. alb and Cry. neo were purchased from ATCC, and other strains
were clinic isolates. C. alb (ATCCY0109) and Cry. neo (ATCCBLS108)
were used as the quality-controlled strains and tested in each
assay. Intermediate 5 and compound B1 that served as the nega-
tive control were synthesized by us, while FLC, ICZ, and VCZ that
served as the positive control were obtained from their respective
manufacturers.
Chemistry
The general synthetic methodology for the preparation of title com-
pounds (6–37) is outlined in Scheme 1. As a key intermediate of
our designed triazole antifungals, the oxirane compound 4 was syn-
thesized by the reported procedure (16). The intermediate 5 was
synthesized by ring-open reaction of oxirane 4 with 4-piperidinol.
The title compounds were synthesized, and the good yield was
obtained when the reaction was performed in CH2Cl2 in the pres-
ence of DMAP (4-dimethylaminopyridine) and EDCI (1-ethyl-3-(3-dim-
ethylaminopropyl) carbodiimide HCl). All the new compounds (6–37)
described previously were characterized by IR, API-ES, and NMR
spectroscopic analysis.
The procedure for the synthesis of the compound
5 1-[2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-
triazol-1-yl)-propyl]-piperidin-4-ol (5)
To a stirred mixture of 1-[2-(2,4-difluorophenyl)-2,3-epoxypropyl]-1H-
1,2,4-triazole methanesulfonate (4) (1.65 g, 0.005 mol), C2H5OH
(30 mL), and N(C2H5)3 (3 mL), 4-piperidinol (0.70 g, 0.006 mol) was
The antifungal potency was evaluated by means of the minimal inhibi-
tory concentration (MICs), using the serial test in the broth microdilu-
tion modification method published by the National Committee for
Clinical Laboratory Standard (NCCLS) method M27-A2 (17). The MIC80
was defined as the first well with an approximate 80% reduction in
growth compared with the growth of the drug-free well. For assays,
Figure 2: Structure and the modified position of the designed compound.
Chem Biol Drug Des 2012; 80: 382–387
383

Chai et al.
Scheme 1: Conditions: (a)
ClCH2COCl, AlCl3, 50 ꢀC, 5 h, in
87% yield; (b) C6H5CH3, NaHCO3,
1H-1,2,4-triazole, reflux, 5 h, in
47% yield; (c) C6H5CH3, (CH3)3SOI,
NaOH, centylmethylammonium bro-
mide, 60 ꢀC, 3 h, in 56% yield; (d)
CH3SO3H, 0 ꢀC, 1 h, in 89% yield;
(e) CH3CH2OH, Et3N, 4-piperidinol,
reflux, 6 h, in 78% yield; (f) various
acids, DMAP, EDCI, CH2Cl2, reflux,
8 h, in 67–81% yield.
the title compounds to be tested were dissolved in dimethyl sulfoxide
(DMSO), serially diluted in growth medium, and inoculated and incu-
bated at 35 ꢀC. Growth MIC was determined at 24 h for C. alb and at
72 h for Cry. neo. The results of assays are summarized in Table 1.
The data point from the mean of replicates. All of our susceptibility
tests were performed three times by each antifungal agent. After the
antifungal test, none of the tested compounds were changed or
hydrolyzed, which were detected by TLC, 1H NMR, and LC-MS.
spectrum against all the tested fungal pathogens. The minimal
inhibitory concentration (MIC80) value indicated that the title com-
pounds showed excellent antifungal activities against C. alb, which
is the most common cause of life-threatening fungal infections.
Most of the compounds were more potent than compound B1,
even more potent than FCZ and ICZ with their MIC80 value in the
range of 0.0039–0.0625 lg ⁄ mL. Noticeably, the MIC80 value of
compound 6, 9, 11, 12, 13, 17, 18, 23, 26, and 27 was 64 times
lower than that of compound B1 (with the MIC80 value of
0.25 lg ⁄ mL) against C. alb in vitro and even the same as the posi-
tive control ICZ (with the MIC80 value of 0.0039 lg ⁄ mL). Moreover,
these compounds also showed excellent inhibitory activity against
other Candida spp., such as C. tro and C. par, with their MIC80
value in the range of 0.0039–1 lg ⁄ mL (compound B1 only showed
0.25 lg ⁄ mL against the two fungi). Cry. neo has a worldwide distri-
bution and is the most common cause of life-threatening fungal
infections. The inhibitory activity of the title compounds against
Cry. neo was superior to that of compound B1. The MIC80 value of
compound 28 (with the MIC80 value of 0.0039 lg ⁄ mL) was 1025
times higher than that of compound B1. Compound B1 was not
effective against Aspergillus fumigates (A. fum), while our com-
pounds showed moderate activity. The MIC80 value of compounds
11, 12, 18, 27, and 28 against A. fum was 0.25 lg ⁄ mL. More-
over, the designed compounds also showed good activity against
dermatophytes (T. rub and M. gyp). For example, compounds 10
and 28 (with the MIC80 value of 0.0156 lg ⁄ mL) were 256 fold
more potent against M. gyp than compound B1.
Protocol of docking study
Based on the previous study (18), we constructed and modified the
3D model of CACYP51. All the molecular modeling calculations
were performed using SYBYL 6.9 version (Tripos, St. Louis, MO, USA).
The structures of the compounds were assigned with Gasteiger–
Hꢀckle partial atomic charges. Energy minimization was performed
using the Tripos force field, Powell optimization method, and MAXI-
MIN2 minimizer with a convergence criterion of 0.001 Kcal ⁄ mol.
Simulated annealing was then performed. The system was heated
to 1000 K for 1.0 ps and then annealed to 250 K for 1.5 ps. The
annealing function was exponential; 50 such cycles of annealing
were run, and the resulting 50 conformers were optimized using
methods described previously. The lowest energy conformation was
selected. All the other parameters were default values.
Results and Discussions
In vitro antifungal activity assay (Table 1) indicates that all the syn-
thesized compounds (6–37) showed excellent activity and broad
Compared with the lead compound B1, the title compounds exhibit
excellent antifungal activity against C. alb. Maybe the oxygen atom
384
Chem Biol Drug Des 2012; 80: 382–387

Triazole as Potential Antifungal Agent
Table 1: Antifungal activities of the title compounds in vitro (MIC80, lg ⁄ mL)
Compound
C. alb Y0109
C. par 0306392
C. tro
C. neo BLS108
F. com
T. rub 0501124
M. gyp 0310388
A. fum 0504656
5
6
8
4
16
64
32
16
8
>64
1
1
1
1
0.0039
0.0156
0.0156
0.0039
0.0156
0.0039
0.0039
0.0039
0.0625
0.0625
0.0156
0.0039
0.0039
0.0156
0.0156
0.0625
0.0625
0.0039
0.0156
0.0625
0.0039
0.0039
0.0625
0.0156
0.0625
0.0156
0.0625
0.0625
0.0625
0.0625
0.0625
0.0156
0.25
0.0156
0.0156
0.0625
0.0039
0.0625
0.0156
0.0156
0.0156
0.0625
0.0625
0.0156
0.0156
0.0039
0.25
0.0156
0.0625
0.0625
0.0156
0.0625
0.0156
0.0156
0.0625
0.0625
0.25
0.0625
0.0156
0.0156
0.0625
0.0625
0.25
0.0625
0.0625
0.0625
0.0156
1
0.0625
0.0156
0.25
0.0625
0.0625
0.0625
0.0625
0.0625
0.25
0.0625
1
0.25
0.0625
0.25
0.25
0.25
0.0625
1
0.25
0.25
0.0625
0.25
0.25
1
0.25
0.0625
0.0625
0.25
1
0.25
0.25
1
0.0625
0.0625
0.25
0.0625
0.0156
0.0625
0.0625
0.0625
0.25
0.25
0.25
0.0625
0.0625
0.25
0.25
1
0.25
0.0625
0.0625
1
7
8
9
0.0625
0.25
0.0625
0.0625
0.0625
0.25
0.0625
0.25
0.0625
0.25
0.0625
1
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
B1
FCZ
ICZ
VCZ
4
0.25
0.25
4
>64
1
4
16
0.25
1
1
1
4
1
4
16
4
0.25
0.25
1
1
1
1
4
4
1
0.0625
0.25
0.25
1
1
1
0.0625
0.0625
0.0625
0.25
0.25
0.0625
0.25
1
0.25
0.0156
0.0625
0.25
0.25
1
0.25
0.0625
0.0625
1
0.0625
0.0625
0.0625
0.25
0.0625
0.25
0.25
0.25
1
0.0156
0.0625
0.0156
0.0625
0.0039
0.0039
0.0625
0.0625
0.25
1
1
1
1
0.25
0.25
0.25
0.25
0.25
0.0156
0.25
0.25
0.25
0.0156
0.0039
0.0625
0.0625
0.25
0.25
0.25
1
0.25
0.0156
0.0039
0.0625
0.0625
0.0156
0.25
1
1
0.25
0.25
0.0625
0.0156
0.0625
0.0625
0.25
0.25
0.25
0.25
0.25
0.25
0.25
4
1
0.25
0.0156
1
1
4
1
0.25
1
0.25
4
0.25
0.0625
0.25
4
0.25
4
16
0.25
0.0625
1
1
>64
>64
1
0.25
4
1
0.25
0.25
0.0625
0.0039
1
4
0.0625
0.0156
0.0625
0.0156
0.0156
0.25
in the side chains could improve the flexibility of the molecule and
make the side chain more easily lock its proper position. All the
designed compounds esters are structurally flexible functional
groups because rotation about the C–O–C bonds has a low barrier.
Their flexibility and low polarity is manifested in their physical prop-
erties. Esters are more polar than ethers but less polar than alco-
hols. They participate in hydrogen bonds as hydrogen-bond
acceptors. As the hydrogen bond and hydrophobic interactions of
the ester, the title compounds can enhance the binding capacity
with the active site of CYP51. As the hydrogen-bond acceptor, the
title compounds esters have a good water solubility. It will be ben-
eficial for the development of new drugs.
to the R configuration of the compounds. The docking results
revealed that the compound 12 binded to the active site of
CACYP51 through the formation of a coordination bond with iron of
heme group. The difluorophenyl group was located in the hydropho-
bic binding cleft lined with Phe126, Leu121, Tyr118, Met306, and
Gly307. The side chain of the compound 12 was oriented into sub-
strate access channel 2 (FG loop) and formed hydrophobic and van
der waals interactions with Ala117, Tyr118, Phe228, Pro230,
Leu376, Ile379, Phe380, and Met508. Moreover, the phenyl group
attached to the piperidinol of the side chain interacted with the
phenol group of Phe380 through the formation of p-p face-to-edge
interaction. There are some differences between the molecular
docking result of compound B1 (Figure 4) and compound 12. The
difluorophenyl group of compound B1 could not locate the common
hydrophobic binding cleft. Although the phenyl group of the side
chain could interact with the phenol group of Phe380 through the
formation of p-p face-to-edge interaction, the side chain could not
embed into substrate access channel 2 (FG loop). All these affect
the combination of compound B1 and the enzyme.
To validate the hypothesis, compound 12 was docked into the
active site of CACYP51 (Figure 3). Literature reported that the chiral
compounds were docked into the active site. The R isomer showed
lower interaction energy with CYP51 than the S isomer, which indi-
cated that the R isomer might have better antifungal activity than
the S isomer (19). In our study, all the docked conformations refer
Chem Biol Drug Des 2012; 80: 382–387
385

Chai et al.
ogenic fungi, which were often encountered clinically. Compared
with the lead compound B1, our present results clearly showed
that the replacement of the piperazinyl side chain with the piperidi-
nol side chain greatly enhanced the antifungal activity of these title
compounds against Candida species and broadened the antifungal
spectrum. This observation was explained by a molecular model
resulting from the computational docking simulation. The side chain
of the compound 12 was oriented into substrate access channel 2
(FG loop) and formed hydrophobic and van der waals interactions
with surrounding hydrophobic residues. The phenyl group of the
side chain can interact with the phenyl group of Phe380 through
the formation of p-p face-to-edge interaction. Further evaluations
are necessary to determine the antifungal activities of these title
compounds in vivo and help us to optimize these new leading com-
pounds, which are continuing in our laboratories.
Figure 3: The binding mode of compound 12 in the active site
of CACYP51.
Acknowledgments
This work was supported by the National Natural Science Founda-
tion of China (Grant No. 30300437), the Eleventh Five Year Military
Medicine and Public Health Research Projects (Grant No.
06MB206), and by Shanghai Leading Academic Discipline Project
(Project No. B906).
References
1. Fridkin S.K., Jarvis W.R. (1996) Epidemiology of nosocomial fun-
gal infections. Clin Microbiol Rev;9:499–511.
2. Wingard J.R., Leather H. (2004) A new era of antifungal therapy.
Biol Blood Marrow Transplant;10:73–90.
3. Dismukes W.E. (2006) Antifungal therapy: lessons learned over
the past 27 years. Clin Infect Dis;42:1289–1296.
4. Goossens H. (2005) European status of resistance in nosocomial
infections. Chemotherapy;51:177–181.
5. Heidemann H.T., Gerkens F.F., Spickard W.A., Jackson E.K.,
Branch R.A. (1983) Amph-otericin B nephrotoxicity in humans
decreased by salt repletion. Am J Med;75:476–481.
6. Garcia-Rodriguez L.A., Duque A., Castellsague J., Pꢁrez-Gutthann
S., Stricker B. H. (1999) A cohort study on the risk of acute liver
injury among users of ketoconazole and other antifungal drugs.
Br J Clin Pharmacol;48:847–852.
7. Lavrijsen A. P., Balmus K. J., Nugteren-Huying W. M., Roldaan
A. C., Van't Wout J. W., Stricker B. H. (1992) Hepatic injury
associated with itraconazole. Lancet;340:251–252.
8. Georgopapadakou N.H., Walsh T.J. (1996) Antifungal agents:
chemotherapeutic targets and immunologic strategies. Antimic-
rob Agents Chemother;40:279–291.
Figure 4: The binding mode of compound B1 in the active site
of CACYP51.
Besides, the negative control intermediate 5 showed poor activity.
After forming ester, the activity of the title compounds was greatly
improved. That might be because the short side chain could not ori-
ent into the substrate access channel 2. Besides, the intermediate 5
could not form p-p face-to-edge interaction. Meanwhile, compound
31–35 showed less activity than others. Furthermore, the longer the
alkyl side chain was, the poorer the activity was. That is because the
long side chain might not be oriented into the substrate access chan-
nel properly. It can also affect the interaction of the other groups
with the target enzyme. So the phenyl group of compound 31–35
cannot form the p-p face-to-edge interaction with Phe380.
9. Chai X. Y., Zhang J., Yu S. C., Hu H. G., Zou Y., Zhao Q. J., Dan
Z. G., Zhang D. Z., Wu Q. Y. (2009) Design, synthesis, and bio-
logical evaluation of novel 1-(1H-1,2,4-triazole-1-yl)-2-(2,4-difluor-
ophenyl)-3- substituted benzylam-ino-2-propanols. Bioorg Med
Chem Lett;19:1811–1814.
10. Guan Z.J., Chai X.Y., Yu S.C., Hu H.G., Jiang Y.Y., Meng Q.G.,
Wu Q.Y. (2010) Synthesis, molecular docking and biological eval-
uation of novel triazole derivatives as antifungal agents. Chem
Biol Drug Des;76:496–504.
Conclusions
In summary, a series of triazole antifungal agents were synthesized,
and their antifungal activities were screened for eight human path-
386
Chem Biol Drug Des 2012; 80: 382–387

Triazole as Potential Antifungal Agent
11. Yu S.C., Chai X.Y., Hu H.G., Yan Y.Z., Guan Z.J., Zou Y., Sun Q.Y.,
Wu Q.Y. (2010) Synthesis and antifungal evaluation of novel
triazole derivatives as inhibitors of cytochrome P450 14a-de-
methylase. Eur J Med Chem;45:4435–4445.
12. Chai X.Y., Zhang J., Cao Y.B., Zou Y., Wu Q.Y., Zhang D.Z., Jiang
Y.Y., Sun Q.Y. (2011) New azoles with antifungal activity: design,
synthesis, and molecular docking. Bioorg Med Chem
Lett;21:686–689.
17. National Committee for Clinical Laboratory Standards (2002) Ref-
erence Method for Broth Dilution Antifungal Susceptibility Test-
ing of Yeasts Approved Standard. Document M27-A2. Wayne,
PA: National Committee for Clinical Laboratory Standards.
18. Ji H. T., Zhang W. N., Zhou Y. J., Zhang M., Zhu J., Song Y. L.,
Lꢀ J. G., Zhu J. (2000) A three-dimensional model of lanosterol
14a-demthylase of Candida albicans and its interaction with
azole antifungals. J Med Chem;43:2493–2505.
13. Jiang Y.W., Cao Y.B., Zhang J., Zou Y., Chai X.Y., Hu H.G., Zhao
Q.J., Wu Q.Y., Zhang D.Z., Jiang Y.Y., Sun Q.Y. (2011) Design,
synthesis and antifungal evaluation of 1-(2-(2,4-difluorophenyl)-2-
hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)-1H-1,2,4-triazol-5(4H)-one.
Eur J Med Chem;46:3135–3141.
19. Sheng C.Q., Zhang W.N., Ji H.T., Zhang M., Song Y.L., Xu H.,
Zhu J., Miao Z.Y., Jiang Q.F., Yao J.Z., Zhou Y.J., Zhu J., Lv J.G.
(2006) Structure-based optimization of azole antifungal agents
by CoMFA, CoMSIA, and molecular docking.
Chem;49:2512–2525.
J
Med
14. Chai X.Y., Zhang J., Cao Y.B., Zou Y., Wu Q.Y., Zhang D.Z., Jiang
Y.Y., Sun Q.Y. (2011) Design, synthesis and molecular docking
studies of novel triazole as antifungal agent. Eur J Med
Chem;46:3167–3176.
15. Sheng C.Q., Zhang W.N., Ji H.T., Song Y.L., Zhang M., Zhou Y.J.,
Lv J.G., Zhu J. (2004) Design, synthesis and antifungal activity
of novel triazole derivatives. Chin Chem Lett;15:404–407.
16. Aoyama Y., Yoshida Y., Sonoda Y., Sato Y. (1987) Metabolism of
32-hydroxy-24, 25-dihydrolanosterol by purified cytochrome P-
45014DM from yeast. Evidence for contribution of the cyto-
chrome to whole process of lanosterol 14a-demethylation. J Biol
Chem;262:1239–1243.
Supporting Information
Additional Supporting Information may be found in the online ver-
sion of this article:
Appendix S1. Supplementary material.
Please note: Wiley-Blackwell is not responsible for the content or
functionality of any supporting materials supplied by the authors.
Any queries (other than missing material) should be directed to the
corresponding author for the article.
Chem Biol Drug Des 2012; 80: 382–387
387