Molecular docking, design, synthesis and antifungal activity study of novel triazole derivatives

Article abstract of DOI:10.1016/j.ejmech.2017.10.081

The incidence of life-threatening fungal infections has dramatically increased for decades. In order to develop novel antifungal agents, two series of (2R,3R)-1-(1H-1,2,4-triazol-1-yl)-2-(2,4-difluorophenyl)-3-(N-substitutied)-2-butanols (3a-o, 5a-f, 8a-u), which were analogues of voriconazole, were designed, synthesized and characterized by ¹H NMR, ¹³C NMR and HRMS. The MIC₈₀ values showed that the target compounds 3a-o indicated better activities than fluconazole on three important fungal pathogens except for 3i. Significant activity of compounds 3d, 3k, 3n, 3m and 3o was observed on the Aspergillus fumigatus strain (MIC₈₀ range: 1–0.125 μg/ml). Especially, compound 3k had strong activity to inhibit the growth of ten fungal pathogens. But it didn't exhibit good activity in in vivo value. Molecular docking experiments demonstrated that 3k possessed superior affinity with target enzyme by strong hydrogen bond from morpholine ring.

Full text of DOI:10.1016/j.ejmech.2017.10.081

European Journal of Medicinal Chemistry xxx (2017) 1e7
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Research paper
Molecular docking, design, synthesis and antifungal activity study of
novel triazole derivatives
,
c
,
,
Junqi Wu a 1, Tingjunhong Ni a 1, Xiaoyun Chai a, Ting Wang a, Hongrui Wang a,
Jindong Chen a, Yongsheng Jin a, Dazhi Zhang a, Shichong Yu a *, Yuanying Jiang b
,
, **
a Department of Organic Chemistry, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
b Center for New Drug Research, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
c Student Bridge, Second Military Medical University, Shanghai 200433, China
a r t i c l e i n f o
a b s t r a c t
Article history:
The incidence of life-threatening fungal infections has dramatically increased for decades. In order to
develop novel antifungal agents, two series of (2R,3R)-1-(1H-1,2,4-triazol-1-yl)-2-(2,4-difluorophenyl)-
3-(N-substitutied)-2-butanols (3a-o, 5a-f, 8a-u), which were analogues of voriconazole, were designed,
synthesized and characterized by 1H NMR, 13C NMR and HRMS. The MIC80 values showed that the target
compounds 3a-o indicated better activities than fluconazole on three important fungal pathogens except
for 3i. Significant activity of compounds 3d, 3k, 3n, 3m and 3o was observed on the Aspergillus fumigatus
Received 2 July 2017
Received in revised form
24 October 2017
Accepted 30 October 2017
Available online xxx
strain (MIC80 range: 1e0.125 mg/ml). Especially, compound 3k had strong activity to inhibit the growth of
Keywords:
Molecular docking
Triazole
ten fungal pathogens. But it didn't exhibit good activity in in vivo value. Molecular docking experiments
demonstrated that 3k possessed superior affinity with target enzyme by strong hydrogen bond from
morpholine ring.
Synthesis
© 2017 Elsevier Masson SAS. All rights reserved.
Antifungal activity
CYP51
1. Introduction
from aspergillosis and 450 thousands died per year worldwide [13].
After the first isolation of azole-resistant A. fum in 1989, ICZ resis-
During the last three decades, the increasing morbidity and
emergence of drug-resistance in life-threatening fungal infections
come to be a significant health problem globally, especially among
population with autoimmune diseases, cancer, organ transplants or
AIDS [1e5]. Generally, Candidosis albicans (C. alb.), cryptococcus
neoformans (C. neo.) and Aspergillus fumigatus (A. fum.) are the
major pathogenic agents for systemic fungal infections [6,7]. Azoles
(e.g. Fluconazole(FCZ), Itraconazole(ICZ), Voriconazole(VCZ) and
Posaconazole(PCZ), Fig. 1) are first-line drugs for mycoses, due to its
broad antifungal effect, high potency and low toxicity [8e12].
Noticeably, it is estimated that three million people are suffering
tance and cross-resistant to other azoles are becoming a major
clinical concern for aspergillosis control currently [13,14]. The
antifungal resistant profile of A. fum and other pathogenic asper-
gillus undermined the antifungal efficacy and potentially lead to
treatment failure [15,16]. Therefore, it is urgent to develop novel
antifungal azoles with excellent activity against a variety of clinical
fungal pathogens, especially Aspergillus spp.
Azoles antifungal act by competitive inhibition of the lanosterol
14
a
-demethylase (CYP51), the necessary enzyme for catalyzing the
-methyl in sterol biosynthesis of fungi
oxidative removal of the 14
a
[17]. Consequently, CYP51 has been regarded as a target for mo-
lecular docking and widely applied in the rational design of anti-
fungal compounds [18e24]. To our knowledge, the active site of
CYP51 could be divided into four parts [25,26]: (a) a coordination
bond with iron of the heme group, (b) the hydrophobic region, (c)
the hydrophilic H bonding region, (d) the narrow hydrophobic cleft
formed by the residues in the N-terminus of helix I and helix B0-
meander 1 loop.
Abbreviations: C. alb., Candidosis albican; C. neo., Cryptococcosis neoformans;
A. fum., Aspergillus fumigatus; FCZ, Fluconazole; ICZ, Itraconazole; VCZ, Vor-
iconazole; PCZ, Posaconazole; CYP51, lanosterol 14a-demethylase; iPrOH, iso-
propanol; DMSO, dimethyl sulfoxide; MIC, Minimum inhibitory concentration;
T. rub., Trichophyton rubru; M. gyp., Microsporum gypseu.
* Corresponding author. Tel: þ86-21-8187-228-801.
Prior studies had reported pharmacophores of antifungal tri-
azoles [18e24], which containing a linker was formed through a
two carbon chain between a triazole ring and a dihalophenyl ring.
** Corresponding author. Tel: þ86-21-8187-1357.
E-mail addresses: yuscc1008@163.com (S. Yu), 13761571578@163.com (Y. Jiang).
1
Junqi Wu and Tingjunhong Ni contributed equally to this work.
https://doi.org/10.1016/j.ejmech.2017.10.081
0223-5234/© 2017 Elsevier Masson SAS. All rights reserved.
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J. Wu et al. / European Journal of Medicinal Chemistry xxx (2017) 1e7
Fig. 1. Triazole antifungal agents used in clinical therapy.
In addition, the carbon alpha to the phenyl ring bored a hydroxyl
group. This binding pattern was further examined by our 3D
computer modeling of the interaction between VCZ and CYP51 of
A. fum. according to the Ji et al.’s study [27]. As shown in Fig. 2, the
pyrimidine group of VCZ interacted with HIS374, SER375, ILF376,
LEU503, PHE504, and ILE373 through the hydrophobic cleft. The
residues, including ALA307, HEM503, ALA303, PHE130, VAL135,
THR126, TYR136, and TYR122, were further considered as forming
nonbonding interactions with the triazolyl group, hydroxyl group
and difluorophenyl group.
In order to find effective systemic antifungal compounds with a
broad antifungal spectrum and less potential to develop resistance,
two new series of (2R,3R)-1-(1H-1,2,4-triazole-1-yl)-2-(2,4-
difluorophenyl)-3-(N-substitutied)-2-butanols (Fig. 3) were
designed. While these compounds included all the essential phar-
macophores, the introduction of an additional amine or 1,2,3-
triazole group in side chain was expected to potentially result in
Fig. 3. Generic structure of the designed Voriconazole analogues.
preferable systemic antifungal compounds that are less possible to
develop drug resistance. Amines and 1,2,3-triazoles were consid-
ered as privileged building blocks for the synthesis of bioconjugates
Fig. 2. Computed Voriconazole binding to the heme iron atom at the active site of CYP51.
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J. Wu et al. / European Journal of Medicinal Chemistry xxx (2017) 1e7
3
due to their high stability, selectivity and less adverse reactions
3.2. In vivo antifungal activity
[18,20,28e30]. These structures indicated formidable stable under
basic and acid hydrolysis including oxidative and reductive re-
actions. What's more, we systematically altered the side chain
structure which was oriented to interact with the narrow hydro-
phobic cleft, to study how it may further affect the antifungal
activity.
Compound 3k and FCZ were studied in an infection model of
systemic C. alb. SC5314 in mice. ICR mice, weighing between 18 and
22 g (male, SLAC Lab Animal Ltd), were used in the model, with
each group included ten mice. Firstly, mice were operated intra-
peritoneal injection of iv with 0.4 ml cyclophosphamide (100 mg/
kg, 3 days, qd). Then, mice were administered orally iv with 0.4 ml
of a suspension containing 2 ꢁ 109 CFU/ml of C. alb. In this model,
compound 3k and FCZ (as a suspension in 0.5% carboxymethyl-
cellulose in distilled water) were administered orally 2 h after
infection (1 mg/kg, 7 days, qd). The vehicle control group received
only the vehicle. In vivo efficacy was determined by the mean of
survival days for 20 days after infections. The results were sum-
marized in Fig. 4.
2. Chemistry
In Scheme 1, synthesis of the target compounds 3a-o were
achieved by reacting with intermediate oxirane 1 and primary or
secondary amines (2a-o) in the presence of LiClO4 in isopropanol
(iPrOH) at 120 ꢀC. In order to further investigate the effect of
antifungal activity during various linkers, novel azoles 5a-f were
synthesized on the basis of the nucleophilic substitution reaction
from potential compound 3d. Accordingly, compounds 5a-f were
prepared through 3d reacted with substituted alkyl halides in NaH,
KI and Acetonitrile (Scheme 2). In addition, as shown in Scheme 3,
the ring-opening reaction of oxirane 1 with NaN3 to give key in-
termediate 7. This was subsequently treated with substituted al-
kynes to obtain target compounds 8a-u through Cu-catalyzed
azide-alkyne cycloaddition.
4. Results and discussion
The MIC80 values of compounds 3a-o indicated that most of the
compounds were active against three important fungal pathogens
and some of them demonstrated higher activities than FCZ and
VCZ, especially for A. fum. species. The compounds 3d, 3m, 3n,
(MIC80 ¼ 1
ml) were better than FCZ (MIC80 > 64
VCZ (MIC80 ¼ 0.25 g/ml) on the A. fum. strain. Except for com-
pound 3i, the MIC80 range for C. alb. was 0.5 g/ml to 0.0156 g/ml,
mg/ml) 3o (MIC80 ¼ 0.5
m
g/ml) and 3k (MIC80 ¼ 0.125
mg/
m
g/ml) and 3k was preceded
m
3. Pharmacology
m
m
showing that these compounds were comparable or superior to FCZ
3.1. In vitro antifungal activity
and VCZ. Significant activity of them was also observed on the
C. neo strain (MIC80 range: 0.5e0.0156
compounds 3d and 3k (MIC80 ¼ 0.0156
m
g/ml). Particularly, both of
According to the National Committee for Clinical Laboratory
Standards (NCCLS) recommendations [31,32]. In vitro antifungal
activity of target compounds 3a-o, 5a-f and 8a-u were measured by
means of the minimum inhibitory concentration (MIC). FCZ and
VCZ were included in positive controls to compare with target
compounds. Tested pathogenic fungi were obtained from the
American Type Culture Collection (ATCC). MIC80 was defined as the
first well with an approximate 80% reduction in growth compared
to the growth of the drug-free well. For assays, test compounds and
positive controls were dissolved in dimethyl sulfoxide (DMSO) in
growth medium, inoculated and incubated at 35 ꢀC. The growth
MIC80 was determined at 24 h for C. alb., at 72 h for C. neo. and at
5e7 days for filamentous fungi. These data were the mean of three
replicate tests performed with each test compound and summa-
rized in Table 1 and Table 2.
mg/ml) were superior to VCZ
mg/ml). Two representative compounds 3d and 3k
(MIC80 ¼ 0.0625
presented strong active against several fungal pathogens in Table 2.
The compound 3k was the most active one during the target
compounds in particular. On the C. neo., Trichophyton rubrum (T.
rub.), Microsporum gypseu (M. gyp.) and A. fum. strain, the com-
pound 3k was superior to FCZ and VCZ. It is worth to further
evaluation due to its excellent antifungal activity toward azole-
resistant clinical isolate.
Moreover, the compounds 5a-f, the further structural modifi-
cation of compound 3d, did not indicate the improvement from 3d.
Compounds 5b and 5c showed similar activity with FCZ in three
important fungal pathogens. Only compound 5a demonstrated a
better inhibiting effect in C. alb. (MIC80 ¼ 0.125
mg/ml) and C. neo.
(MIC80 ¼ 0.125 g/ml) than FCZ. The lower activity of Compounds
m
Scheme 1. Synthetic route of the target compounds 3a-o.
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J. Wu et al. / European Journal of Medicinal Chemistry xxx (2017) 1e7
Scheme 2. Synthetic route of the target compounds 5a-f.
Scheme 3. Synthetic route of the target compounds 8a-u.
Table 1
Antifungal activities of the target compounds 3a-o, 5a-f, 7 and 8a-u in vitro. (MIC ¼
m
g/ml).a
Compd
C. abl SC5314
C. neo.
A. fum.
Compd
C. abl SC5314
C. neo.
A. fum.
3a
3b
3c
3d
3e
3f
3g
3h
3i
0.5
0.5
32
8
16
1
4
8
8
4
8
2
0.125
64
1
1
0.5
8
>64
8
8
>64
>64
8a
8b
8c
8d
8e
8f
8g
8h
8i
8j
8k
8l
8m
8n
8o
8p
8q
8r
0.125
0.125
0.5
0.0625
0.0625
0.5
16
0.125
0.25
0.0156
0.125
0.125
0.125
0.125
0.5
0.25
0.03125
2
0.125
0.03125
0.0156
0.125
1
0.0625
0.25
0.0156
0.125
0.125
0.125
0.125
0.5
0.5
0.0156
2
0.125
0.125
0.125
0.125
1
>64
>64
>64
>64
32
>64
32
32
4
8
0.125
0.0625
0.25
0.125
0.125
0.0625
0.0156
0.25
0.5
0.5
0.5
0.0625
0.0625
0.5
0.0625
0.0625
0.0625
0.125
0.0156
0.0625
0.0156
0.25
1
0.5
1
0.0625
0.0625
1
3j
3k
3l
>64
16
>64
>64
>64
>64
16
3m
3n
3o
5a
5b
5c
5d
5e
5f
4
1
8
8
8
2
8
5
4
>64
>64
32
>64
8
8s
8t
8u
7
FCZ
VCZ
1
1
0.0625
0.25
0.0156
1
0.0625
0.25
0.0625
2
FCZ
VCZ
1
2
>64
0.25
>64
0.25
0.03125
0.03125
0.03125
0.03125
Abbreviations: C. alb, Candida albicans; C. neo, Cryptococcus neoformans; A. fum, Aspergillus fumigates. FCZ, fluconazole; VCZ, voriconazole.
a
Minimum inhibitory concentration for 80% inhibition of growth.
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J. Wu et al. / European Journal of Medicinal Chemistry xxx (2017) 1e7
5
Table 2
Antifungal activities of the target compounds 3d and 3k in vitro. (MIC ¼
m
g/ml).a
Compd
C. abl SC5314
C. abl SCY0109
C. gla,
C.neo.
C. kru.
A.fum.
C. tro.
C. par
M. gyp
T. rub
3d
3k
FCZ
VCZ
0.0156
0.03125
1
0.0078
0.0156
0.25
0.0156
0.03125
1
0.0156
0.0156
2
0.5
0.5
32
1
0.5
0.5
32
0.25
0.0625
0.5
0.125
0.0625
8
0.0625
0.0625
1
0.125
>64
0.25
0.03125
0.0156
0.03125
0.0625
0.25
0.25
0.03125
0.25
0.125
Abbreviations: C. alb, Candida albicans; C. neo, Cryptococcus neoformans; C. par, Candida parapsilosis; C. tro, Candida tropicalis; C. kru, Candida krusei; C. gla, Candida glabrata; T. rub,
Trichophyton rubrum; M. gyp, Microsporum gypseu;. A. fum, Aspergillus fumigates. FCZ, fluconazole; VCZ, voriconazole.
a
Minimum inhibitory concentration for 80% inhibition of growth.
placed into the hydrophobic pocket formed by ALA303, ALA307,
VAL135, PHE130, TYR136 and HEM580. Although the hydroxyl
group of compound 3k did not form direct interaction with CYP51,
it might formed hydrogen-bonding interaction with ALA307
mediated by crystal waters. Finally, the morpholine ring could
interact with a hydrophobic pocket built by ILE376, PHE504, ILE373,
SER375 and HIS374. Especially , the strong hydrogen bond existing
between morpholine ring and LEU503 might be the cause of the
effective antifungal activity and broad antifungal spectrum.
Therefore, the side chain was the key pharmacophore, and the
spatial orientations of the pharmacophores were just oriented in
the hydrophobic pocket. The introduction of various side chains
was a vital factor of novel compounds.
Fig. 4. In vivo efficacy of compound 3k and FCZ (1 mg/kg) in a systemic infection of an
ICR mouse model (n ¼ 10) with C. ald. SC5314. *, p < 0.05.
5. Conclusion
5a-f probably associated with reduction of the interaction between
compounds and target enzyme caused by the extension of molec-
ular structure.
Comparing with the intermediate 7, establishment of 1,2,3-
triazole and introduction of phenyl groups or heterocycle groups
In conclusion, two series of novel triazole derivatives with
substituted amines or 1,2,3-triazoles as side chain have been syn-
thesized, and their antifungal activities were evaluated for several
human pathogenic fungi. In vitro biological evaluations of the target
compounds 3d and 3k showed strong antifungal activities. Novel
on position 4 of the 1,2,3-triazole did not result in a significantly
compound 3k could against nearly all the tested fungi, particularly,
increased antifungal activity. The MIC80 range for C. alb. and C. neo.
Aspergillus spp.. However, the in vivo evaluation of the target
were 1 mg/ml to 0.0156 mg/ml after excluding 8d. Furthermore, the
compound 3k indicated it possessed inferior activity in the mice.
The docking result demonstrated the side chain of morpholine ring
could form a closely hydrogen-bonding interaction with CYP51,
which may lead to the discovery of a series of compounds for
further optimization.
various group substituted within this phenyl ring derivatives 8a-o
indicated fluorine and chlorine substituted (8f-k) possessed better
activity than bromine and amino substituted (8i-o). In the other
hand, with the extension of carbons in the para-position, anti-
fungal activity became worse (8a-d). In addition, the unsubstituted
pyridine and thiophene ring analogs (8p-s) were tested and proved
to be comparable to those of the phenyl analogs. During this novel
azoles, compound 8k was the potent antifungal agent to C. alb. and
6. Experimental part
Melting points were measured on a Yamato MP-21 melting
point apparatus. 1H and 13C Nuclear magnetic resonance (NMR)
spectra were achieved in Chloroform-D (CDCl3) unless otherwise
indicated with a Bruker AC-300P spectrometer or a Bruker Avance II
600 spectrometer. Tetramethylsilane (TMS) was considered as the
internal standard. ESI mass spectra were produced on an Aglient
Technologies 6538 UHD Accurate-Mass Q-TOF LC/MS. Silica gel
plates GF254 (Yantai Huanghai chemical, china) were applied to
thin-layer chromatography (TLC) analysis. Amines were purchased
from Energy-Chemical. The solvents and reagents were consumed
from commercial vendors and used as received or dried prior to use
as needed.
C. neo. (MIC80 ¼ 0.0156
mg/ml). However, compounds 8a-u were
confirmed as less effective antifungal activity against A. fum.
As summarized in Fig. 4, the selected compound 3k, which
demonstrated significant in vitro activity as well as broad antifungal
spectrum, was evaluated in an ICR mice model infected by C. alb.
SC5314. The efficacy (survival curve) of orally administered com-
pound 3k less than that of orally administered FCZ at a dose of
1 mg/kg. Perhaps the lack of inferior activity of compound 3k could
be related to its poor Pharmacokinetics.
On the basis of the molecular docking, we constructed a 3D
model of Aspergillus' CYP51. A likely binding mode of compound 3k
in the active site of CYP51 was proposed based on docking result
(Fig. 5). Sheng et al. found that the interaction between isomers (R
and S) of compound and CYP51 was produced by a similar binding
mode. However, S isomer showed higher interaction energy with
CYP51 than the R isomer, which demonstrated that the R isomer
might had better antifungal activity than the S isomer [25]. In our
study, the docked conformation refereed to the R configuration of
the compounds. As shown in Fig. 5, the N(4) atom of the 1,2,4-
triazole moiety coordinated with the heme Fe-atom, meanwhile
the 2,4-difluorophenyl group in the designed compound could be
6.1. General procedure for the synthesis of derivatives 3a-o
To a solution of oxirane 1 (200 mg, 0.796 mmol) in iPrOH (4 ml)
were added LiClO4 (170 mg, 1.592 mmol) and 2a (143 mg,
3.184 mmol). The mixture was stirred continuously for 9 h in an
oven-dried pressure-tight reaction tube and heated at 120 ꢀC. The
solvent was distilled off under reduced pressure. Then, the crude
residue was extracted with dichloromethane (100 ml). The organic
layer was washed with H2O (75 ml ꢁ 2) and brine (75 ml ꢁ 2), dried
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J. Wu et al. / European Journal of Medicinal Chemistry xxx (2017) 1e7
Fig. 5. Computed compound 3k binding to the heme iron atom at the active site of CYP51.
on anhydrous Na2SO4 and filtrated. Then the solvent was evapo-
rated under reduced pressure. The crude residue was purified by
flash chromatography on silica gel (PE: EA 10:1e3:1) to compound
3a as a white solid. Yield: 70.8%; m.p. 100.1e102.9 ꢀC; 1H NMR
procedure as the compound 5a.
6.3. General procedure for the synthesis of derivatives 7
(600 MHz, CDCl3)
d 7.95 (1H, s, triazole-H), 7.74 (1H, s, triazole-H),
A mixture of intermediate oxirane 1 (200 mg, 0.796 mmol),
NaN3 (103 mg, 1.260 mmol), NH4Cl (84 mg, 1.260 mmol) and DMF
(10 mL). The reaction mixture was stirred continuously for 8 h in an
oven-dried pressure-tight reaction tube and heated at 100 ꢀC. The
mixture was extracted with dichloromethane (2 ꢁ 80 mL), the
organic layers were gathered and washed with water (2 ꢁ 100 mL)
and brine (2 ꢁ 100 mL), dried on anhydrous Na2SO4, Then the
solvent was evaporated under reduced pressure. The crude residue
was purified by flash chromatography on silica gel (PE: EA
10:1e5:1) to intermediate 7 as a white solid. Yield: 90.8%; m.p.
7.41e7.37 (1H, m, Ar-H), 6.76e6.73 (2H, s, Ar-H), 4.86e4.72 (2H, m,
CH2), 3.08e3.04 (1H, m, eCH), 2.89e2.84 (1H, m, eCH2), 2.60e2.55
(1H, m, eCH2), 1.12e1.09 (3H, m, eCH2), 0.92 (3H, d,
J ¼ 6.0 Hz,eCH3); 13C NMR (150 MHz, CDCl3) d: 162.03, 158.23,
150.69, 143.69, 129.89, 123.20, 110.87, 103.28, 56.90, 55.55, 42.12,
29.18, 15.21, 15.05; HRMS (ESI) m/z calcd for C14H18F2N4O [MþH]þ:
297.1521, found: 297.1523
The target compounds 3b-o were synthesized by the same
procedure as the compound 3a.
136.7e137.9 ꢀC; 1H NMR (300 MHz, CDCl3)
d 7.80e7.75 (2H, m,
triazole-H), 7.39e7.33 (1H, m, Ar-H), 6.76e6.69 (2H,m, Ar-H),
4.86e4.72 (2H, m, eCH2), 3.78e3.75 (1H, m, eCH), 1.13 (3H, d,
J ¼ 6.0 Hz, eCH3); 13C NMR (75 MHz, CDCl3) d: 162.95, 158.03,
151.93, 143.90, 130.45, 126.460, 111.94, 104.14, 77.80, 59.81, 55.30,
12.98; HRMS (ESI) m/z calcd for C12H12F2N6O [MþH]þ: 295.1113,
found: 295.1112
6.2. General procedure for the synthesis of derivatives 5a-f
The solution of compound 3d (200 mg, 0.649 mmol) in aceto-
nitrile (5 ml) were added NaH (31.2 mg,1.298 mmol) and KI (1.6 mg,
0.009 mmol) followed by iodomethane 4a (184 mg, 1.298 mmol).
The reaction was stirred continuously for 4 h at room temperature.
The mixture was extracted with dichloromethane (2 ꢁ 80 mL), the
organic layers were gathered and washed with water (2 ꢁ 100 mL)
and brine (2 ꢁ 100 mL), dried on anhydrous Na2SO4, filtered and the
solvent was evaporated under reduced pressure. The crude residue
was purified by flash chromatography on silica gel (PE: EA
10:1e2:1) to afford compound 5a as canary yellow solid. Yield:
6.4. General procedure for the synthesis of derivatives 8a-u
A mixture of intermediate 7 (200 mg, 0.680 mmol), phenyl-
acetylene (128 mg, 1.260 mmol) and DMSO (10 mL). The catalyst of
sodium ascorbate (20 mg, 0.100 mmol) and CuSO4$5H2O (25 mg,
0.100 mmol) in H2O (1 mL) was added into the reaction solution
quickly. The mixture was stirred continuously for 2 h, Then the
reaction solution was put into NH3$H2O, extracted with dichloro-
methane (80 mL). The organic layers were gathered and washed
with water (2 ꢁ 150 mL) and brine (2 ꢁ 100 mL), dried on anhy-
drous Na2SO4 and filtered. The solvent was evaporated in a vacuum
and crude residue was purified by flash chromatography on silica
gel (PE: EA 10:1e2:1) to afford compound 8a as a white solid. Yield:
75.4%; m.p. 119.5e120.9 ꢀC; 1H NMR (300 MHz, CDCl3)
d 7.90 (1H, s,
triazole-H), 7.75 (1H, s, triazole-H), 7.30e7.25 (1H, m, Ar-H),
6.87e6.78 (2H, s, Ar-H), 5.06e5.01 (1H, m, eCH2), 4.78e4.73 (1H,
m, eCH2), 3.36e3.31 (1H, m, eCH), 2.38e2.35 (4H, m, eCH3, eCH),
0.88 (3H, d, J ¼ 6.0 Hz, eCH3), 0.51e0.39 (4H, m, eCH2, eCH2); 13
C
NMR (75 MHz, CDCl3) d: 164.26, 160.77, 150.64, 144.40, 131.16,
111.35, 104.60, 83.84, 66.36, 54.99, 53.58, 38.59, 37.30, 8.04, 7.64,
6.92; HRMS (ESI) m/z calcd for C16H20F2N4O [MþH]þ: 323.1678,
found: 323.1678
92.5%; m.p. 104.2e105.1 ꢀC; 1H NMR (600 MHz, CDCl3)
triazole-H), 7.83e7.82 (2H, d, Ar-H), 7.77 (1H, s, triazole-H), 7.66
d 8.15 (1H, s,
The target compounds 5b-f were synthesized by the same
Please cite this article in press as: J. Wu, et al., Molecular docking, design, synthesis and antifungal activity study of novel triazole derivatives,
European Journal of Medicinal Chemistry (2017), https://doi.org/10.1016/j.ejmech.2017.10.081

J. Wu et al. / European Journal of Medicinal Chemistry xxx (2017) 1e7
7
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(1H, s, triazole-H), 7.46e7.38 (3H, m, Ar-H), 7.31e7.29 (1H, m, Ar-H),
6.79e6.74 (2H, m, Ar-H), 5.48 (1H, s, eOH), 5.46e5.44 (1H, m, eCH),
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151.85, 147.95, 144.09, 130.55, 130.40, 128.88, 128.88, 128.31, 125.68,
125.68, 122.20, 119.80, 112.05, 104.42, 77.44, 60.52, 54.77, 15.44;
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397.1582
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Acknowledgements
This work was supported by Chinese natural science fund (No.
81330083, 81673280, 81573473) Creativity and Innovation Training
Program of Second Military Medical University (No. FH2016064,
FH2016065).
Appendix A. Supplementary data
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Please cite this article in press as: J. Wu, et al., Molecular docking, design, synthesis and antifungal activity study of novel triazole derivatives,
European Journal of Medicinal Chemistry (2017), https://doi.org/10.1016/j.ejmech.2017.10.081