Re-engineering of PIP3-antagonist triazole PITENIN's chemical scaffold: Development of novel antifungal leads

Article abstract of DOI:10.1039/c5ra25145a

A novel 4-(1-phenyl-1-hydroxyethyl)-1-(o-hydroxyphenyl)-1H-1,2,3-triazole was designed by integrating the structural features of triazole PITENIN anticancer agents and the azole class of antifungal drugs. A two-step protocol comprising the Barbier proparg

Full text of DOI:10.1039/c5ra25145a

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ARTICDLOIE: 10.10T39/YC5RPA251E45A
Re-engineering of PIP3-Antagonist triazole PITENIN’s chemical
scaffold: development of novel antifungal leads†
Sravani Pulya,a Yadagiri Kommagalla,a Duhita G. Sant,b Shweta U. Jorwekar,b Santosh G. Tupe,*b
Mukund V. Deshpande*b and Chepuri V. Ramana*a
5
Received (in XXX, XXX) XthXXXXXXXXX 20XX, Accepted Xth XXXXXXXXX 20XX
DOI: 10.1039/b000000x
angiogenic therapies.6 Similarly, recent Phase II studies have
A
novel 4-(1-phenyl-1-hydroxyethyl)-1-(o-hydroxyphenyl)-
50 revealed the use of antifungal azole drug itraconazole for
advanced prostate cancer treatment.7 We have also developed a
new class of anticancer agents which interfere with the interaction
between phosphatidylinositol (3,4,5)-trisphosphate (PIP3) and
Pleckstrin homology (PH) domains and are termed PITENIN
55 analogs (PIT-1, PIT-2 and DM-PIT-1 having acylthiourea unit)
and subsequent investigations have led to the identification of
1,4-disubstituted 1,2,3-triazoles (Triazole PIT) as promising
anticancer agents in vivo and in vitro.8 Moreover, we have also
been working on the development of novel antifungal agents
60 employing the 1,2,3-triazole moiety as an alternative
pharmacophore in the search for a new azole class of antifungal
drugs.9
1H-1,2,3-triazole was designed by integrating the structural
features of triazole PITENIN anticancer agents and azole
10 class of antifungal drugs. A two-step protocol comprising the
Barbier propargylation and Cu-catalyzed azide-alkyne
cycloaddition was established to synthesise a diverse set of
compounds of this class. Their screening against a wide range
of human fungal pathogens led to identification of several
15 potential antifungal hits and some of them displayed better
antifungal activity than fluconazole against Candida glabrata,
Cryptococcus neoformans, Aspergillus fumigatus and
Aspergillus niger. Mode of action studies revealed that their
antifungal activity was resulting either from the inhibition of
20 lanosterol 14 α-demethylase enzyme (leading to ergosterol
depletion) or by the generation of reactive oxygen species
(ROS).
1. Introduction
Fungal infections cause a continuous and momentous threat to
25 human health, especially in immuno-compromised patients. Drug
resistance among fungal pathogens is an increasing problem,
hence there is continuing demand for safe and effective broad
spectrum anti-fungal agents.1 Among the several conventional
anti-fungal agents used, azoles are the most frequently employed
30 antifungal drugs, due to their high therapeutic index, broad
spectrum of activity and more favourable safety profile.2
Structurally, one of the most important groups of azole
antifungals is triazole alcohols. Fluconazole,
a
bis-
triazolylpropan-2-ol derivative was the first triazole alcohol
35 antifungal drug which was introduced in to the market.3 It has
initiated remarkable progress in the treatment of candidiasis. Its
widespread use has limited its clinical efficacy. Hence, the
replacement of 1,2,4-triazole with 1,2,3-triazole as well as the
incorporation of several functional groups on the triazole ring
40 have provided
a much extended chemical and biological
functionalities on the triazole scaffold.4 In recent years, studies
have revealed that many of the cancer drugs have the potential to
be antifungal agents conversely, antifungal compounds also show
potential anticancer properties.5 For example, Thiabendazole, an
45 orally available antifungal drug in clinical use for 40 years, which
also potently inhibits angiogenesis in animal models and in
human cells and thus acts as a potential complementary
therapeutic agent for use in combination with current anti-
Figure 1: Structures of (i) (Triazole-)PITENIN anticancer agents; (ii)
selected antifungal drugs Fluconazole, Ravuconazole and
Voriconazole indicating structural elements A (red box) and B (green
box); and (iii) novel anticancer pharmacophore integrating the
substructures A and B present in various azole class of antifungals.
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Page 2 of 12
were synthesized by the traditional Cu(I)-catalyzed azide alkyne
35 cycloaddition approach employing CuSO4-sodium ascorbate-
2. Results and Discussion
View Article Online
proline combination in tBuOH-water as solvent.12 The
DOI: 10.1039/C5RA25145A
2.1. Synthesis of compounds
cycloaddition reactions proceeded smoothly at rt within 5–6 h
and provided the requisite 1,2,3-triazoles in very good yields. The
structures of all the final compounds were confirmed by 1H
40 NMR, 13C NMR, and HRMS methods and the purity was
determined (> 98%) with the help of analytical HPLC.
Having these newly developed 1,2,3-triazole anticancer hits in
hand, we reasoned that a slight structural/functional group
modification of Triazole PIT by incorporating some of the key
aspects of 1,2,4-triazole antifungals, shall lead to new anti-fungal
compounds. Figure 1 reveals the salient features of our design. A
dissection of the 1,2,4-traizole antifungals revealed two key
5
2.2. Antifungal screenings
2.2.1. Antifungal activity of the triazoles. Next, we proceeded
for the screening of these novel azoles against different human
45 pathogenic yeasts and filamentous fungi.13 The minimum
inhibitory concentrations (MIC) of all the compounds against
tested human fungal pathogens are presented in Table 1. Many of
these compounds exhibited potent antifungal activity against
several of the tested pathogens. Compounds 3kb, 3kd were most
50 potent against all the fungal pathogens, with MIC in the range of
4–32 μg/mL. Compounds 3lb, 3mb, 3la, 3ma and 3da also
showed very good antifungal activity except against A. niger
(MIC >128 μg/mL). Compounds 3kc, 3id, 3ha and 3ga showed
moderate antifungal activity. Among the pathogens, A. niger was
55 least susceptible with an MIC of >128 μg/mL for 12 out of 35
compounds. Ten compounds exhibited antifungal activity against
filamentous Aspergilli, for which fluconazole was not effective
(MIC >128 μg/mL). All the compounds showed better activity
than fluconazole against Candida glabrata NCYC 388. The
60 minimum fungicidal concentration (MFC) of selected active
compounds was also estimated. It can be noted from Table 2 that
compounds 3kb, 3kd and 3ia exerted strong fungicidal effect
with MFC values being the same as MIC or 2 X MIC. Whereas,
for compound 3ja complete killing was not observed (MFC >256
65 μg/mL) indicating its fungi-static action.
10 central structural elements A (N-N-C-C-OH) and B (N=C-C-C-
OH). A novel hybrid triazole scaffold was then designed by
incorporating the key central structural units in the Azole PIT
scaffold.10
Scheme 1. Synthesis of alkynol precursors
O
Br
Zn, THF
HO
R
R
R'
R'
sat. NH4Cl
rt, 2.5 h
OH
nBu
HO
HO
HO
1a (65%)
1b (62%)
1c (67%)
1d (72%)
HO
HO
HO CF3
HO Ph
F
Cl
1e (70%)
1f (71%)
1g (69%)
1h (74%)
R
HO
HO
HO
HO
F3C
Br
F
F
Cl
Cl
1k (73%)
CF3
1i (64%)
1j (71%)
1l (R=H, 63%)
1m (R=CH3, 67%)
Coming to the structure activity relationship, the antifungal
activity was not significant when there was no substituent on the
left side aromatic ring for both the 4-chloro and 5-chloro series.
The presence of a single halo-substituent on this aryl ring, as in
70 the case of the triazoles 3ga, 3ha, 3ia, 3gb, 3hb, 3ib, was seen to
lead to a moderate improvement in the antifungal activity.
Interestingly, when two chlorine/fluorine atoms were placed on
this ring, as in the case of 3jb and 3kb, it resulted in significant
enhancement in the activity for the 5-chloro series. Similarly, the
75 compounds 3la, 3ma, 3lb and 3mb, where there are two
trifluromethyl groups present on this aromatic motif (that are
important in case of the PITENIN analogues), showed excellent
antifungal activity. The addition of dichloro or bis-trifluoromethyl
groups on the scaffold is expected to increase the lipophilicity
80 and also strongly polarize the parent molecule. In the azido part
of the structure, as evident from the MIC values, the activity
improved from no substitution<5-fluoro<4-chloro<5-chloro with
respect to –OH. Based on MIC values exhibited, the compounds
3da, 3kb, 3kd, 3la, 3lb and 3ma were identified as promising
85 molecules (Figure 2). After having identified these potential hits,
we proceeded for finding their mode of action.
15 With this idea, we proceeded further for the synthesis of a
collection of compounds having the designed hybrid triazole
scaffol. The synthesis of these triazoles has been planned from
the Cu-catalyzed [3+2]-cycloaddition of a suitably functionalized
1-aryl-but-3-yn-1-ol (1) with a 2-azidophenol (2). A large set of
20 alkynol intermediates (1a–1m) having functional group diversity
on aryl rings and also varying the substitution such as –CH3, –Et,
-nBu, –Ph and finally –CF3 at the central hydroxyl bearing 3°-
carbon have been synthesized by the Zn-mediated Barbier
reaction using propargyl bromide (Scheme 1).11
25 In case of anticancer DM-PIT analogues and Triazole PIT hits,
the best activities were seen when the N-aryl group brings a
chlorine substituent along with the o-OH group.8 Considering
this, 2-azido-4/5-chlorophenols (2a/2b) have been selected as the
key azidophenols partners for building the library along with 2-
30 azido-phenol (2c) and 2-azido-5-fluorophenol (2d) for preparing
additional series. These 2-azidophenols 2a–2d have been
synthesized from the corresponding 2-aminophenols following
established procedures. The targeted 1,2,3-triazole derivatives 1
2|
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Table 1: Minimium Inhibitory Concentration (MIC90) of triazoles (in µg/mL) against different human pathogenic fungi.
C. albicans
C .albicans
C. neoformans
C. glabrata
A. niger
A. fumigatus
S. cerevisiae ATCC 201390
Compound
ATCC 24433
ATCC 10231
ATCC 34664
NCYC 388
ATCC 10578
NCIM 902
wild strain
erg11 mutant
3aa
3ba
3ca
3da
3ea
64
128
64
16
64
4
128
128
32
16
>128
32
32
8
32
32
>128
16
16
128
128
32
>128
64
128
16
16
32
16
>128
>128
32
16
64
64
64
32
32
128
128
32
8
64
64
64
16
16
32
128
8
64
64
128
>128
>128
32
64
64
64
32
>128
>128
>128
>128
128
64
>128
64
128
64
128
128
32
128
>128
128
32
64
32
32
128
64
>128
128
32
128
>128
64
32
64
8
>128
64
64
64
64
>128
>128
>128
32
128
128
128
>128
64
64
64
64
64
16
128
8
>128
128
128
128
32
3fa
16
16
3ga
3ha
3ia
3ja
3ka
3la
3ma
3ab
3bb
3cb
3eb
3fb
3gb
3hb
3ib
3jb
3kb
3lb
3mb
3gc
3hc
3ic
32
16
32
32
64
16
16
64
>128
32
64
32
32
32
16
16
4
16
16
128
16
32
64
8
128
64
>128
128
16
64
>128
16
16
8
32
16
>128
128
32
32
16
64
32
32
32
128
64
64
64
64
16
32
32
32
16
8
128
128
64
32
16
128
128
64
32
16
16
16
128
128
128
16
32
16
16
128
32
128
64
32
64
64
128
32
32
128
128
128
16
32
16
32
64
16
8
16
8
32
16
>128
>128
>128
128
128
>128
64
>128
128
64
8
>128
128
128
128
32
>128
128
16
>128
128
128
128
32
>128
64
64
128
32
>128
64
16
64
32
64
64
16
>128
16
128
128
128
64
32
64
64
128
32
3jc
3kc
3gd
3hd
3id
128
128
8
128
16
2
>128
64
64
128
64
>128
3jd
128
8
8
128
16
32
>128
32
>128
3kd
Fluconazole
16
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µg/mL) for the mutant as compared to the wild type.
View Article Online
Table 2: Minimum fungicidal concentration (MFC) of the selected
compounds (in μg/mL)
DOI: 10.1039/C5RA25145A
2.2.3. Effect of the compounds on sterol profile of Candida
35 albicans ATCC 24433. The inhibition of lanosterol 14 α-
demethylase by azoles lead to the depletion of ergosterol and the
accumulation of methylated sterol intermediates such as eburicol
and obtusifoliol.16 In spectrophotometric sterols analysis, the
presence of ergosterol and the late sterol intermediate
40 24(28)dehydroergosterol (24(28)DHE) in a sample without any
compound resulted in a characteristic four-peaked curve. In the
presence of 3la, 3ma, 3kb, 3kd and fluconazole, a dose-
dependent decrease in the height of the absorbance peaks was
observed and corresponded to the decrease in the ergosterol
45 concentration in C. albicans ATCC 24433 cells (Figure 3, data
shown for 3kb and fluconazole only). The % ergosterol decrease
was calculated and depicted in Figure 4. The results indicated that
compounds 3la, 3ma, 3kb and 3kd are potent inhibitors of
lanosterol 14 α-demethylase, and that they exerted their
50 antifungal action through ergosterol depletion. No ergosterol
depletion was observed for 3da and 3lb indicating a different
mode of action. As ergosterol is the major sterol component of
the fungal cell membrane Inhibition of lanosterol 14 α-
demethylase and subsequent ergosterol depletion by the
55 compounds leads to loss in cell integrity and function.
Organism
3fa
3kb
3kd
3ia
3ja
3jb
C. albicans
ATCC 24433
C. albicans
ATCC 10231
C. neoformans
ATCC 34664
C. glabrata
NCYC 388
A. niger
128
16
32
32
>256
>256
128
128
64
32
32
32
64
32
64
64
64
32
>256
>256
>256
>256
>256
>256
64
32
32
64
256
256
256
32
256
64
64
ATCC 10578
A. fumigatus
NCIM 902
64
Figure 2. Selected triazoles for their mode of action studies
2.2.2. Haploinsufficiency assay for identification of ergosterol
synthesis inhibitors. yeast genome-wide drug induced
A
5
haploinsufficiency screen using Saccharomyces cerevisiae as a
model organism has been developed by Giaever et al.14 The
screen employs a diploid S. cerevisiae mutant strain which has
only one copy of a specific gene, whereas there are two copies of
all other genes in it. The resulting heterozygote mutant strain
10 becomes sensitive to any drug that acts on the product of this
gene as compared to the wild type strain. This haploinsufficient
phenotype thereby identifies the gene product of the heterozygous
locus as the likely drug target.15 In the present study, a diploid
wild type S. cerevisiae strain and its various mutants (including
15 mutants haploid for the genes erg11 – lanosterol 14 α-
demethylase; CHS2 – chitin synthase; FAB1 - fructose 1,6-bis-
phosphate aldolase; FAS2 – Fatty acid synthase; ILV5 –
Acedohydroxyacid reductoisomerase etc.) were used for the
screening of the lead compounds. For instance, in case of mutant
20 with only one copy of erg11, the effect of azole compounds
(those having lanosterol 14 α-demethylase enzyme as the target)
on growth will be more pronounced on it as compared to the wild
type, whereas other agents will exert similar effect on both the
strains. All the synthesized compounds were checked for
25 antifungal activity against S. cerevisiae wild and mutant strains.
For all the mutants tested except for erg11, the MIC values of all
compounds were similar to the MICs against S. cerevisiae wild
strain (data not shown). In case of erg11 mutant, the
concentration of compounds 3fa, 3ga, 3la and 3ma required to
30 inhibit its growth was half than the MIC for wild type S.
cerevisiae suggesting lanosterol 14 α-demethylase as their target
(Table 1). Standard fluconazole also exhibited half MIC (16
Figure 3. Spectrophotometric sterol analysis of C. albicans ATCC 24433
exposed with different concentrations of compound 3kb and fluconazole.
Figure 4. Depletion of total ergosterol content of C. albicans
ATCC24433 exposed to 8 µg/ml (open bars) and 16 µg/ml (filled bars)
60 concentrations of compounds.
4|
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is difficult to determine which events lead to loss of viability of
2.2.4. Detection of Reactive Oxygen Species (ROS)
cells following damage.19 ROS are also implicated in apoptosis
generation. Apart from lanosterol 14 α-demethylase inhibition,
few azoles like miconazole are known to exert their antifungal
action by reactive oxygen species (ROS) generation.17 In fungal
cells, mitochondria are the source of reactive oxygen species
(ROS). Chemical compounds may be involved in the release of
intracellular ATP and the inhibition of membrane ATPase
activity leading to ROS production.17 Therefore, ROS production
by the lead compounds in C. albicans ATCC 24433 was
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and necrotic death. It is one of the important hallmarks of
DOI: 10.1039/C5RA25145A
40 apoptosis in which DNA fragmentation, chromatin condensation,
externalisation of phosphatidyl serine, membrane damage takes
place.
5
2.2.5. Haemolysis assay. Cellular toxicity of the compounds was
checked by haemolysis assay.20 The concentrations tested were in
45 the range of 4–512 μg/mL. None of the compounds except 3ma
caused haemolysis at the concentrations tested. Even for 3ma, the
haemolysis was observed at much higher concentrations than
MIC i.e. 3.3% and 37.4% at 256 and 512 μg/mL, respectively.
10 evaluated by dichlorofluorescin diacetate (DCFH-DA) staining.
DCFH-DA is a non-fluorescent dye that undergoes intracellular
deacetylation followed by ROS mediated oxidation to
a
fluorescent 2′,7′-dichlorofluorescein and is generally used to
measure ROS generation in the cytoplasm and cellular
15 organelles.18
50 3. Conclusions
In conclusion, founded upon anticancer leads and integrating the
structural features of azole anti-fungal drugs, a novel 4-(1-
phenyl-1hydroxyethyl)-1-(o-hydroxyphenyl)-1H-1,2,3-triazole
was designed and further led to synthesis of corresponding series
55 of compounds employing various functional group modifications
around it. The antifungal activities and mode of action of the
compounds were also studied. From a total of 35 synthesized
compounds, six compounds (3da, 3kb, 3kd, 3la, 3lb and 3ma)
showed promising antifungal activity against human pathogenic
60 fungi. Some of these new entities exhibited better antifungal
activity than fluconazole against C. glabrata, C. neoformans, A.
fumigatus and A. niger. Mode of action studies revealed that
compounds 3ma and 3la inhibited the lanosterol 14 α-
demethylase enzyme, thus exerting antifungal action through
65 ergosterol depletion. Compounds 3lb and 3da caused fungal
death through ROS generation. In case of compounds 3kb and
3kd, generation of ROS and associated ergosterol depletion led to
the death of the fungal pathogen. The observed structure-activity
relationship will be useful in lead optimization. With simple
70 two/three-step synthesis, high overall yield, purity and ease of
functional group diversification, the new series offers an
attractive template for lead discovery. This series of compounds
hold promise for the control of pathogenic fungi and need to be
further investigated for increasing activity and therapeutic
75 potential.
Figure 5. DCFH-DA staining of Candida albicans ATCC 24433 cells
exposed to A) 32 µg/ml 3kd; B) 8 µg/ml 3kb; A1 and B1 - respective
20 bright field images.
100
90
80
70
60
50
40
30
20
10
0
4. Experimental
4.1. General experimental procedure
All the cycloaddition reactions were carried out employing 100
mg of the alkyne. A representative procedure as follows: to a
80 solution of alkyne 1a (100 mg, 0.68 mmol) and azide 2a (116 mg,
t
3lb
3da
3kb
3kd
0.68 mmol) in BuOH:H2O (4 mL, 3:1) at rt, sodium ascorbate
Figure 6. Reactive oxygen species generation in C. albicans ATCC
24433 exposed to MIC (open bars) and 2 X MIC (filled bars) of
25 compounds.
(97 mg, 0.59 mmol) and CuSO4·5H2O (31 mg, 0.14 mmol) were
added and the resulting brick reddish mixture was stirred
vigorously for 6 h. The reaction mixture was diluted with water
85 (5 mL) and extracted with EtOAc (2 x 10 mL). The organic layer
was dried (Na2SO4), concentrated under reduced pressure. The
crude product was purified by column chromatography with their
respective solvent systems.
The results indicated that 3lb and 3da caused fungal death
through ROS generation. ROS generation was not observed for
3la and 3ma, while compounds 3kb and 3kd apart from
ergosterol inhibition also showed ROS generation in C. albicans
30 ATCC 2443 (Figure 5). The level of ROS production increased in
a dose-dependent manner and >80% fluorescent cells were
observed at 2X MIC for all the compounds except for 3da
(Figure 6). The fungicidal nature of 3kb and 3kd was confirmed
by ROS generation which is in correlation with the MFC values.
35 ROS can damage a wide range of molecules, including nucleic
acids, proteins and lipids, and with these wide range of targets, it
4.1.1.
4-Chloro-2-(4-(2-hydroxy-2-phenylethyl)-1H-1,2,3-
90 triazol-1-yl)phenol (3aa). 178 mg (83% yield); Rf = 0.3
(petroleum ether/EtOAc = 7:3); Colourless solid; Mp: 148–150
°C; 1H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 3.22 (d, J = 5.3
Hz, 2H), 5.04 (t, J = 6.6 Hz, 1H), 7.03 (d, J = 8.8 Hz, 1H), 7.22–
7.43 (m, 6H), 7.71 (d, J = 2.5 Hz, 1H), 8.08 (s, 1H) ppm; 13C
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NMR [50 MHz, CDCl3 + MeOH (D4)]: δ 34.8 (t), 72.5 (d), 117.7
(d), 123.3 (d), 123.8 (s), 123.9 (d), 124.6 (s), 125.3 (d, 2C), 126.9
4.1.7.
4-Chloro-2-(4-(2-(4-fluorophenyl)-2-hydroxypropyl)-
1H-1,2,3-triazol-1-yl)phenol (3ga). 154 mg (79% yield); Rf =
View Article Online
DOI: 10.1039/C5RA25145A
(d), 127.8 (d, 2C), 129.0 (d), 143.2 (s), 143.7 (s), 147.4 (s) ppm; 65 0.3 (petroleum ether/EtOAc = 7:3); Colourless solid; Mp: 164–
HRMS (ESI) calcd for C16H15O2N3Cl (M++H): 316.0847; found:
316.0847.
166 °C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.6 (s, 3H),
3.21 (s, 2H), 6.90−6.99 (m, 3H), 7.17 (dd, J = 8.8, 2.5 Hz,
1H),7.33−7.42 (m, 2H),7.59 (d, J = 2.5 Hz, 1H), 7.77 (s, 1H); 13C
NMR [100 MHz, CDCl3 + MeOH (D4)]: δ 29.0 (q), 39.9 (t), 73.3
70 (s), 114.2 (d), 114.7 (d), 118.3 (d), 123.1 (d), 123.8, 124.3 (s),
126.4 (d, 2C), 126.5 (d, 2C), 129.3 (d, 2C),142.7 (s), 147.5 (s)
ppm; HRMS (ESI+): calcd. For C17H16ClFN3O2(M++H):
348.0910; found 348.0904.
1
5
4.1.2.
4-Chloro-2-(4-(2-hydroxy-2-phenylpropyl)-1H-1,2,3-
triazol-1-yl)phenol (3ba). 164 mg (79% yield); Rf = 0.4
(petroleum ether/EtOAc = 7:3); Colourless solid; Mp: 180–181
1
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.61 (s, 3H),
10 3.28 (s, 2H), 6.98 (d, J = 8.8 Hz, 1H), 7.20–7.37 (m, 4H), 7.43–
7.49 (m, 2H), 7.65 (d, J = 2.5 Hz, 1H), 7.83 (s, 1H) ppm; 13C
NMR [50 MHz, CDCl3 + MeOH (D4)]: δ 27.9 (q), 39.5 (t), 73.0
(s), 117.6 (d), 123.3 (d), 123.7 (s), 124.3 (d, 3C), 124.5 (s), 126.0
(d), 127.4 (d, 2C), 128.9 (d), 142.9 (s), 146.7 (s), 147.4 (s) ppm;
15 HRMS (ESI) calcd for C17H17O2N3Cl (M++H): 330.1004; found:
330.0999.
4.1.8.
4-Chloro-2-(4-(2-(4-chlorophenyl)-2-hydroxypropyl)-
75 1H-1,2,3-triazol-1-yl)phenol (3ha). 154 mg (82% yield); Rf =
0.4 (petroleum ether/EtOAc = 7:3); Orange color solid; Mp: 171–
173 °C; H NMR [400 MHz, CDCl3 + MeOH (D4)]: δ 1.54 (s,
3H), 3.21 (s, 2H), 6.94 (d, J = 8.8 Hz, 1H), 7.17 (dd, J = 8.6, 2.5
Hz, 1H), 7.23 (d, J = 8.6 Hz, 2H), 7.32−7.38 (m, 3H), 7.59 (d, J =
80 2.5 Hz, 1H), 7.81 (br. s., 1H); 13C NMR [100 MHz, CDCl3 +
MeOH (D4)]: δ 29.0 (q), 39.6 (t), 73.3 (s), 118.3 (d), 123.2 (d),
123.9 (s), 124.3 (s), 124.6 (s), 126.3 (d, 2C), 127.9 (d, 3C), 129.3
(d), 132.2 (s), 145.6 (s), 147.6 (s) ppm; HRMS (ESI+): calcd. For
C17H16Cl2N3O2(M++H): 364.0614; found 364.0609.
1
4.1.3.
4-Chloro-2-(4-(2-hydroxy-2-phenylbutyl)-1H-1,2,3-
triazol-1-yl)phenol (3ca). 168 mg (85% yield); Rf = 0.4
(petroleum ether/EtOAc = 7:3); Colourless solid; Mp: 153–155
20 °C; 1H NMR [400 MHz, CDCl3 + MeOH (D4)]: δ 0.80 (t, J = 7.0
Hz, 3H), 1.85–2.02 (m, 2H), 3.32 (s, 2H), 6.97 (d, J = 8.0 Hz,
1H), 7.18–7.22 (m, 2H), 7.31 (t, J = 7.3 Hz, 2H), 7.37–7.42 (m,
2H), 7.57 (s, 1H), 7.71 (s, 1H) ppm; 13C NMR [50 MHz, CDCl3 +
MeOH (D4)]: δ 7.5 (q), 34.7 (t), 38.4 (t), 76.5 (s), 118.7 (d), 122.8
25 (d), 123.4 (d), 124.4 (s, 2C), 125.3 (d, 2C), 126.4 (d), 127.9 (d,
2C), 129.4 (d), 143.5 (s), 144.7 (s), 147.7 (s) ppm; HRMS (ESI)
calcd for C18H19O2N3Cl (M++H): 344.1160; found: 344.1164.
85 4.1.9.
2-(4-(2-(4-Bromophenyl)-2-hydroxypropyl)-1H-1,2,3-
triazol-1-yl)-4-chlorophenol (3ia). 142 mg (83% yield); Rf = 0.3
(petroleum ether/EtOAc = 7:3); Brown color solid; Mp: 183–185
1
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.52 (s, 3H),
3.20 (s, 2H), 6.93 (d, J = 8.8 Hz, 1H), 7.11−7.42 (m, 5H), 7.52 (d,
90 J = 2.5 Hz, 1H), 7.77 (s, 1H); 13C NMR [50 MHz, CDCl3
+
4.1.4.
4-Chloro-2-(4-(2-hydroxy-2-phenylhexyl)-1H-1,2,3-
MeOH (D4)]: δ 29.3 (q), 39.6 (t), 73.5 (s), 118.8 (d), 120.5 (s),
122.7 (d), 123.4 (d), 124.4 (s), 124.5 (s), 126.7 (d, 2C), 129.4 (d),
131.0 (d, 2C), 146.2 (s, 2C), 147.6 (s) ppm; HRMS (ESI+): calcd.
For C17H16BrClN3O2 [M++H]: 408.0109; found: 408.0102; calcd.
95 For C17H16Br37ClN3O2 [M++H]: 410.0079; found: 410.0076;
calcd. For C17H1681Br37ClN3O2 [M++H]: 412.0059; found:
412.0043
triazol-1-yl) phenol (3da). 158 mg (86% yield); Rf = 0.5
30 (petroleum ether/EtOAc = 6:4); Colourless solid; Mp: 165–167
°C; 1H NMR [400 MHz, CDCl3 + MeOH (D4)]: δ 0.83 (t, J = 7.3
Hz, 3H), 1.01–1.11 (m, 1H), 1.20–1.36 (m, 3H), 1.81–1.89 (m,
1H), 1.91–1.99 (m, 1H), 3.32 (t, J = 15.3 Hz, 1H), 4.03 (bs, 2H),
6.98 (d, J = 8.8 Hz, 1H), 7.19–7.23 (m, 2H), 7.32 (t, J = 7.8 Hz,
35 1H), 7.40 (d, J = 7.5 Hz, 2H), 7.58 (d, J = 2.3 Hz, 1H), 7.69 (s,
1H) ppm; 13C NMR [100 MHz, CDCl3 + MeOH (D4)]: δ13.4 (q),
22.6 (t), 25.2 (t), 38.7 (t), 41.7 (t), 76.0 (s), 118.2 (d), 123.2 (d),
123.9 (d), 124.1 (s), 124.6 (s), 125.1 (d, 2C), 126.1 (d), 127.7 (d,
2C), 129.2 (d), 143.2 (s), 145.0 (s), 147.6 (s) ppm; HRMS (ESI)
40 calcd for C20H23O2N3Cl (M++H): 372.1473; found: 372.1471.
4.1.10.
4-Chloro-2-(4-(2-(2,4-difluorophenyl)-2-hydroxyl
propyl)-1H-1,2,3-triazol-1-yl)phenol (3ja). 154 mg (82% yield);
100 Rf = 0.5 (petroleum ether/EtOAc = 7:3); Brown color solid; Mp:
1
191–193 °C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.60
(d, J = 1.0 Hz, 3H), 3.26 (d, J = 14.9 Hz, 1H), 3.44 (d, J = 14.8
Hz, 1H), 6.64−6.78 (m, 2H), 6.93 (d, J = 8.8 Hz, 1H), 7.15 (dd, J
= 8.8, 2.5 Hz, 1H), 7.40−7.54 (m, 2H), 7.76 (s, 1H); 13C NMR
105 [125 MHz, CDCl3 + MeOH (D4)]: δ 27.4 (q), 37.4 (t), 72.1 (s),
103.5 (dd, J = 25.7, 27.7 Hz), 110.1 (dd, J = 2.9, 20.0 Hz), 117.8
(d), 123.2 (d), 124.1 (t),124.58 (s), 128.2 (dd, J = 6.5, 9.5 Hz),
129.3 (s), 143.0 (s), 147.4 (s), 157.8 (s), 159.8 (s), 160.7 (s),
4.1.5. 4-Chloro-2-(4-(2-hydroxy-2,2-diphenylethyl)-1H-1,2,3-
triazol-1-yl)phenol (3ea). 147 mg (83% yield); Rf = 0.5
(petroleum ether/EtOAc = 7:3); Colourless solid; Mp: 194–196
1
°C; H NMR [200 MHz, CDCl3]: δ 3.82 (s, 2H), 6.96 (d, J = 8.7
45 Hz, 1H), 7.17–7.35 (m, 7H), 7.46–7.51 (m, 6H), 7.58 (d, J = 2.5
Hz, 1H), 7.66 (s, 1H) ppm; 13C NMR [50 MHz, CDCl3]: δ37.9
(t), 77.2 (s), 118.0 (d), 123.3 (d), 124.0 (s), 124.2 (d), 124.5 (s),
125.8 (d, 4C), 126.5 (d, 2C), 127.6 (d, 4C), 129.1 (d), 143.1 (s),
146.0 (s, 2C), 147.5 (s) ppm; HRMS (ESI) calcd. for
50 C22H19O2N3Cl (M++H): 392.1160; found: 392.1162.
162.6
(s)
ppm;
HRMS
(ESI+):
calcd.
For
110 C17H15ClF2N3O2(M++H): 366.0815; found 366.0816.
4.1.11. 4-Chloro-2-(4-(2-(2,4-dichlorophenyl)-2-hydroxy
propyl)-1H-1,2,3-triazol-1-yl)phenol (3ka). 142 mg (81%
yield); Rf = 0.3 (petroleum ether/EtOAc = 6:4); Orange color
solid; Mp: 178–180 °C; H NMR [500 MHz, CDCl3 + MeOH
4.1.6.
4-Chloro-2-(4-(3,3,3-trifluoro-2-hydroxy-2-
1
phenylpropyl)-1H-1,2,3-triazol-1-yl)phenol (3fa). 154 mg
115 (D4)]: δ 1.92 (s, 3H), 3.64 (d, J = 14.9 Hz, 1H), 3.87 (d, J = 14.9
Hz, 1H), 7.10 (dd, J = 8.9, 2.1 Hz, 1H), 7.20 (d, J = 2.1 Hz, 1H),
7.35 (dd, J = 8.5, 2.1 Hz, 1H), 7.53 (d, J = 2.1 Hz, 1H), 7.71 (d, J
= 8.5 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 8.09 (br. s., 1H); 13C
NMR [125 MHz, CDCl3 + MeOH (D4)]: δ 26.1 (q), 35.7 (t), 73.4
120 (s), 116.5 (d), 119.3 (d), 122.9 (s), 124.6 (d, 2C), 126.2 (d), 128.9
(d, 2C), 129.9 (d), 130.7 (s), 132.6 (s), 134.4 (s), 142.0 (s), 149.5
(s) ppm; HRMS (ESI+): calcd. For C17H15Cl3N3O2 (M++H):
398.0224; found 398.0224.
(86% yield); Rf = 0.4 (petroleum ether/EtOAc = 7:3); Colourless
solid; Mp: 202–206 °C; H NMR [400 MHz, CDCl3 + MeOH
1
55 (D4)]: δ 3.57 (d, J = 15.1 Hz, 1H), 3.77 (d, J = 15.1 Hz, 1H), 6.92
(td, J = 7.7, 1.1 Hz, 1H), 6.98 (dd, J = 8.2, 1.4 Hz, 1H), 7.23–7.38
(m, 4H), 7.50 (dd, J = 8.0, 1.6 Hz, 1H), 7.61 (d, J = 7.33 Hz, 2H),
7.76 (s, 1H); 13C NMR [100 MHz, CDCl3 + MeOH (D4)]: δ 33.1
(t), 118.1 (d), 121.1 (s), 125.7 (s), 126.0 (s), 126.7 (s), 128.3 (s),
60 129.2 (s), 129.5 (s), 131.3 (s), 138.2 (s), 142.2 (s), 150.8 (s) ppm;
HRMS (ESI) calcd for C17H14O2N3ClF3 (M++H): 384.0721;
found: 384.0720.
6|
Journal Name, [year], [vol], 00–00
This journal is © The Royal Society of Chemistry [year]

Page 7 of 12
RSC Advances
(petroleum ether/EtOAc = 7:3); Colourless solid; Mp: 210–212
4.1.12.
2-(4-(2-(3,5-Bis(trifluoromethyl)phenyl)-2-hydroxy
1
ethyl)-1H-1,2,3-triazol-1-yl)-4-chlorophenol (3la). 139 mg
(87% yield); Rf = 0.4 (petroleum ether/EtOAc = 7:3); Colourless
solid ; Mp: 181–183 °C; H NMR [200 MHz, CDCl3 + MeOH
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 3.81 (s, 2H),
View Article Online
6.89 (dd, J = 2.2, 8.6 Hz, 1H), 7.02 (m, 1H), 7.15–7.33 (m, 6H),
1
DOI: 10.1039/C5RA25145A
7.38 (d, J = 1.6 Hz, 1H), 7.43–7.50 (m, 4H), 7.54 (s, 1H) ppm;
5
(D4)]: δ 3.22 (d, J = 6.4 Hz, 2H), 5.19 (t, J = 6.4 Hz, 1H), 7.02 (d, 65 13C NMR [50 MHz, CDCl3 + MeOH (D4)]: δ 38.0 (t), 77.2 (s),
J = 8.8 Hz, 1H), 7.26 (dd, J = 2.7, 8.6 Hz, 1H), 7.76 (d, J = 2.7
Hz, 1H), 7.79 (bs, 2H), 7.90 (bs, 2H), 8.17 (s, 1H) ppm; 13C
NMR [50 MHz, CDCl3 + MeOH (D4)]: δ 34.8 (t), 72.1 (d), 117.7
(d), 120.5 (s), 120.6 (d, J = 4.0 Hz), 123.5 (d, 2C), 123.9 (s, 2C),
117.3 (d), 119.8 (d), 122.8 (s), 123.9 (d), 124.2 (d), 125.9 (d, 4C),
126.7 (d, 2C), 127.8 (d, 4C), 134.7 (s), 143.2 (s), 146.0 (s, 2C),
149.7 (s) ppm; HRMS (ESI) calcd for C22H19O2N3Cl (M++H):
392.1160; found: 392.1158.
10 124.2 (d), 124.6 (s), 125.7 (d, J = 2.9 Hz), 129.1 (d), 130.6 (s, d, J
= 33.3 Hz ), 131.6 (s, d, J = 33.3 Hz), 142.8 (s), 146.7 (s), 147.5
(s) ppm; HRMS (ESI) calcd for C18H13O2N3ClF6 (M++H):
452.0595; found: 452.0600.
70 4.1.18.
5-Chloro-2-(4-(3,3,3-trifluoro-2-hydroxy-2-
phenylpropyl)-1H-1,2,3-triazol-1-yl)phenol (3fb). 148 mg
(82% yield); Rf = 0.4 (petroleum ether/EtOAc = 7:3); Colorless
solid; Mp: 203–205 °C; H NMR [400 MHz, CDCl3 + MeOH
1
(D4)]: δ 3.56 (d, J = 15.6 Hz, 1H), 3.71 (d, J = 15.6 Hz, 1H), 6.88
75 (dd, J = 8.5, 2.1 Hz, 1H), 6.97 (d, J = 2.3 Hz, 1H), 7.24−7.34 (m,
3H), 7.47 (d, J = 8.2 Hz, 1H), 7.57 (d, J = 7.8 Hz, 2H), 7.7 (s,
1H); 13C NMR [100 MHz, CDCl3 + MeOH (D4)]: δ 32.8 (t),
117.9 (d), 120.7 (s), 124.3 (s), 125.9 (s), 126.1 (s), 127.7 (s),
128.9 (s), 129.2 (s), 135.9 (s), 137.5 (s), 141.8 (s), 150.9 (s) ppm;
80 HRMS (ESI) calcd for C17H14O2N3ClF3 (M++H): 384.0721;
found: 384.0720.
4.1.13.
2-(4-(2-(3,5-Bis(trifluoromethyl)phenyl)-2-hydroxy
15 propyl)-1H-1,2,3-triazol-1-yl)-4-chlorophenol (3ma). 134 mg
(85% yield); Rf = 0.4 (petroleum ether/EtOAc = 7:3); Colourless
solid; Mp: 164–166 °C; H NMR [200 MHz, CDCl3 + MeOH
1
(D4)]: δ 1.67 (s, 3H), 3.30 (s, 2H), 7.00 (d, J = 8.8 Hz, 1H), 7.24
(dd, J = 2.5, 8.8 Hz, 1H), 7.69 (d, J = 2.5 Hz, 1H), 7.76 (bs, 1H),
20 7.97 (s, 2H), 8.08 (s, 1H) ppm; 13C NMR [50 MHz, CDCl3 +
MeOH (D4)]: δ28.1 (q), 39.4 (t), 72.7 (s), 117.7 (d), 119.9 (d, J =
4.0 Hz), 120.3 (s), 123.4 (d, 2C), 123.9 (s, 2C), 124.5 (d), 125.1
(d, d, J = 2.9 Hz), 125.7 (s), 129.1 (d), 130.7 (s, d, J = 32.9 Hz),
143.2 (s, 2C), 147.7 (s) 150.3 (s) ppm; HRMS (ESI) calcd for
25 C19H15O2N3ClF6 (M++H): 466.0752; found: 466.0755.
4.1.19. 5-Chloro-2-(4-(2-(4-fluorophenyl)-2-hydroxypropyl)-
1H-1,2,3-triazol-1-yl)phenol (3gb). 157 mg (87% yield); Rf =
0.4 (petroleum ether/EtOAc = 7:3); Orange color solid; Mp: 176–
1
85 178 °C; H NMR [400 MHz, CDCl3 + MeOH (D4)]: δ 1.55 (s,
3H), 3.20 (s, 2H), 6.87 (d, J = 2.2 Hz, 1H), 6.90−6.92 (m, 1H),
6.94 (s, 1H), 6.97−7.00 (m, 2H), 7.32−7.40 (m, 2H), 7.46 (d, J =
4.1.14.
5-Chloro-2-(4-(2-hydroxy-2-phenylethyl)-1H-1,2,3-
triazol-1-yl)phenol (3ab). 178 mg (83% yield); Rf = 0.3
8.6 Hz, 1H), 7.69 (br. s., 1H); 13C NMR [50 MHz, CDCl3
+
(petroleum ether/EtOAc = 7:3); Colourless solid; Mp: 171–172
1
MeOH (D4)]: δ 24.6 (q), 35.3 (t), 68.7 (s), 109.7 (d, J = 21.2 Hz,
90 2C), 112.8 (d), 115.3 (d), 119.6 (d, 2C), 121.8 (d, J = 8.1 Hz, 2C),
130.2 (s), 138.2 (s), 145.1 (s, 2C), 154.4 (s), 161.9 (s) ppm;
HRMS (ESI+): calcd. For C17H16ClFN3O2 (M++H): 348.0910;
found 348.0906.
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 3.19 (s, 2H),
30 5.01 (s, 1H), 6.95 (d, J = 8.3 Hz, 1H), 7.06 (s, 1H), 7.25–7.36 (m,
5H), 7.58 (d, J = 8.0 Hz, 1H), 8.01 (s, 1H) ppm; 13C NMR [50
MHz, CDCl3 + MeOH (D4)]: δ 34.7 (t), 72.4 (d), 116.4 (d), 119.3
(d), 122.9 (s), 124.7 (d), 125.2 (d, 2C), 126.8 (d), 127.6 (d, 3C),
134.4 (s, 2C), 143.2 (s), 149.5 (s) ppm; HRMS (ESI) calcd. for
35 C16H15O2N3Cl (M++H): 316.0847; found: 316.0854.
4.1.20. 5-Chloro-2-(4-(2-(4-chlorophenyl)-2-hydroxypropyl)-
95 1H-1,2,3-triazol-1-yl)phenol (3hb). 158 mg (83% yield); Rf =
0.3 (petroleum ether/EtOAc = 7:3); Orange color solid; Mp: 202–
4.1.15. 5-Chloro-2-(4-(2-hydroxy-2-phenylpropyl)-1H-1,2,3-
triazol-1-yl)phenol (3bb)
1
204 °C; H NMR [400 MHz, CDCl3 + MeOH (D4)]: δ 1.53 (s,
3H), 3.20 (s, 2H), 6.89 (dd, J = 8.7, 2.1 Hz, 1H), 7.00 (d, J = 2.2
171 mg (83% yield); Rf = 0.4 (petroleum ether/EtOAc = 7:3);
Colourless solid; Mp: 189–190 °C; 1H NMR [200 MHz, CDCl3 +
40 MeOH (D4)]:δ 1.61 (s, 3H), 3.28 (s, 2H), 6.95 (dd, J = 2.3, 8.6
Hz, 1H), 7.05 (d, J = 2.3 Hz, 1H), 7.22–7.36 (m, 3H), 7.44–7.49
(m, 2H), 7.55 (d, J = 8.6 Hz, 1H), 7.76 (s, 1H) ppm; 13C NMR
[50 MHz, CDCl3 + MeOH (D4)]: δ 28.2 (q), 39.6 (t), 73.1 (s),
116.6 (d), 119.4 (d), 122.9 (s), 124.4 (d, 3C), 124.7 (d), 126.1 (d),
45 127.5 (d, 2C), 134.5 (s), 142.9 (s), 146.7 (s), 149.6 (s) ppm;
HRMS (ESI) calcd for C17H17O2N3Cl (M++H): 330.1004; found:
330.1009.
Hz, 1H), 7.22 (m, J = 8.8 Hz, 2H), 7.30−7.36 (m, 2H), 7.45 (d, J
100 = 8.6 Hz, 1H), 7.72 (s, 1H); 13C NMR [100 MHz, CDCl3
+
MeOH (D4)]: δ 29.2 (q), 39.7 (t), 73.4 (s), 117.5 (d), 120.0 (d),
122.9 (s), 124.2 (d), 126.3 (d, 2C), 128.0 (d, 3C), 132.3 (s), 134.8
(s), 145.7 (s, 2C), 149.7 (s) ppm; HRMS (ESI+): calcd. For
C17H16Cl2N3O2 (M++H): 364.0614; found 364.0610.
1054.1.21.
2-(4-(2-(4-Bromophenyl)-2-hydroxypropyl)-1H-1,2,3-
triazol-1-yl)-5-chlorophenol (3ib). 138 mg (81% yield); Rf = 0.5
(petroleum ether/EtOAc = 7:3); Brown color solid; Mp: 196–198
1
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.53 (s, 3H),
3.20 (s, 2H), 6.89 (dd, J = 8.6, 2.2 Hz, 1H), 7.00 (d, J = 2.2 Hz,
110 1H), 7.26−7.33 (m, 2H), 7.36 (s, 1H), 7.40 (m, 1H), 7.47 (d, J =
8.6 Hz, 1H), 7.74 (s, 1H); 13C NMR [50 MHz, CDCl3 + MeOH
(D4)]: δ 29.3 (q), 39.6 (t), 73.5 (s), 117.7 (d), 120.1 (d), 120.5 (s),
123.9 (d), 126.7 (d, 2C), 131.0 (d, 2C), 134.9 (s), 146.2 (s, 2C),
149.7 (s, 2C), 154.7 (s) ppm; HRMS (ESI+): calcd. For
115 C17H16BrClN3O2 (M++H): 408.0109; found 408.0098; calcd. For
C17H16Br37ClN3O2 (M++H): 410.0079; found; 410.0073; calcd.
For C17H1681Br37ClN3O2 (M++H): 412.0059; found: 412.0051.
4.1.16.
5-Chloro-2-(4-(2-hydroxy-2-phenylbutyl)-1H-1,2,3-
triazol-1-yl)phenol (3cb). 158 mg (80% yield); Rf = 0.4
50 (petroleum ether/EtOAc = 6:4); Yellow solid; Mp: 129–131 °C;
1H NMR [400 MHz, CDCl3 + MeOH (D4)]: δ 0.79 (t, J = 7.4 Hz,
3H), 1.83–2.05 (m, 2H), 3.32 (s, 2H), 6.88 (dd, J = 8.5, 2.3 Hz,
1H), 7.05 (d, J = 2.3 Hz, 1H), 7.18–7.22 (m, 1H), 7.28–7.34 (m,
3H), 7.38–7.40 (m, 2H), 7.57 (bs, 1H) ppm; 13C NMR [50 MHz,
55 CDCl3 + MeOH (D4)]: δ 7.8 (q), 35.1 (t), 38.5 (t), 76.7 (s), 118.6
(d), 120.2 (d), 122.0 (d), 125.4 (d, 3C), 126.6 (d), 128.1 (d, 2C),
134.8 (s, 2C), 144.7 (s, 2C), 149.8 (s) ppm; HRMS (ESI) calcd
for C18H19O2N3Cl (M++H): 344.1161; found: 344.1164.
4.1.22.
5-Chloro-2-(4-(2-(2,4-difluorophenyl)-2-hydroxy
propyl)-1H-1,2,3-triazol-1-yl)phenol (3jb). 151 mg (81%
4.1.17. 5-Chloro-2-(4-(2-hydroxy-2,2-diphenylethyl)-1H-1,2,3-
60 triazol-1-yl)phenol (3eb). 143 mg (81% yield); Rf = 0.5
120 yield); Rf = 0.4 (petroleum ether/EtOAc = 7:3); Brown color
1
solid; Mp: 187–189 °C; H NMR [400 MHz, CDCl3 + MeOH
This journal is © The Royal Society of Chemistry [year]
Journal Name, [year], [vol], 00–00 |7

RSC Advances
Page 8 of 12
(D4)]: δ 1.59 (s, 3H), 3.26 (d, J = 14.9 Hz, 1H), 3.41 (d, J = 14.9
(petroleum ether/EtOAc = 7:3); Orange color solid; Mp: 159–161
1
°C; H NMR [500 MHz, CDCl3 + MeOH (D4)]: δ 1.56 (br. s.,
Hz, 1H), 6.65−6.74 (m, 2H), 6.85 (dd, J = 8.6, 2.1 Hz, 1H), 6.98
(d, J = 1.9 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.42−7.49 (m, 1H),
View Article Online
3H), 3.22 (br. s., 2H), 6.89−6.97 (m, 1H), 7.00 (d, J = 7.6 Hz,
7.73 (s, 1H) ppm; 13C NMR [125 MHz, CDCl3 + MeOH (D4)]: δ 65 1H), 7.20−7.31 (m, 3H), 7.38 (d, J = 7.3 Hz, D2OHI):,170..150439(d/C, 5JR=A275.1045A
5
28.2 (q), 37.4 (t), 72.8 (s), 103.7 (s), 103.9 (dd, J = 26.1, 27.7
Hz), 110.6 (s), 110.7 (dd, J = 3.4, 20.20 Hz), 117.8 (d), 120.1 (d),
122.6 (s), 123.0 (d), 123.6 (d), 128.5 (s), 134.8 (s), 143.6 (s),
149.7 (s), 157.9 (s), 159.8 (s), 160.9 (s), 162.9 (s) ppm; HRMS
Hz, 1H), 7.83 (br. s., 1H); 13C NMR [125 MHz, CDCl3 + MeOH
(D4)]: δ 28.4 (q), 39.6 (t), 72.9 (s), 116.7 (d), 119.4 (d), 123.6 (d),
123.9 (s), 124.2 (d), 126.1 (d, 2C), 127.6 (d, 2C), 129.5 (d), 131.9
(s), 142.7 (s), 145.6 (s, 2C), 148.9 (s) ppm; HRMS (ESI+): calcd.
(ESI+): calcd. For C17H15ClF2N3O2(M++H): 366.0815; found 70 For C17H17ClN3O2 (M++H): 330.1004; found 330.0998.
10 366.0814.
4.1.28. 2-(4-(2-(4-Bromophenyl)-2-hydroxypropyl)-1H-1,2,3-
triazol-1-yl)phenol (3ic). 132 mg (84% yield); Rf = 0.3
(petroleum ether/EtOAc = 7:3); Brown color solid; Mp: 164–166
4.1.23.
5-Chloro-2-(4-(2-(2,4-dichlorophenyl)-2-hydroxy
propyl) -1H-1,2,3-triazol-1-yl)phenol (3kb). 145 mg (83%
yield); Rf = 0.5 (petroleum ether/EtOAc = 7:3); Orange color
solid; Mp: 192–194 °C; H NMR [500 MHz, CDCl3 + MeOH
1
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.53 (s, 3H),
1
75 3.21 (s, 2H), 6.86−6.96 (m, 1H), 6.97−7.04 (m, 1H), 7.16−7.24
(m, 1H), 7.27−7.33 (m, 2H), 7.34−7.46 (m, 3H), 7.71 (s, 1H); 13
C
15 (D4)]: δ 1.68 (s, 3H), 3.34 (d, J = 15.1 Hz, 1H), 3.78 (d, J = 14.7
Hz, 1H), 6.9 (dd, J = 8.7, 2.3 Hz, 1H), 6.97 (d, J = 2.3 Hz, 1H),
7.09 (dd, J = 8.7, 2.3 Hz, 1H), 7.26 (d, J = 2.3 Hz, 1H), 7.37 (d, J
= 8.7 Hz, 1H), 7.61 (d, J = 8.7 Hz, 1H), 7.71 (s, 1H); 13C NMR
[100 MHz, CDCl3 + MeOH (D4)]: δ 27.0 (q), 35.9 (t), 74.3 (s),
20 117.8 (d), 120.1 (d), 122.7 (s), 123.6 (d), 126.9 (d), 129.4 (d),
130.5 (d), 130.9 (s), 133.2 (s), 134.8 (s), 142.0 (s), 149.7 (s) ppm;
HRMS (ESI+) calcd for C17H15O2N3ClF6 (M++H): 398.0224;
found: 398.0225.
NMR [50 MHz, CDCl3 + MeOH (D4)]: δ 29.3 (q), 39.6 (t), 73.4
(s), 117.8 (d), 120.0 (d), 120.4 (s), 122.6 (d), 123.1 (d), 126.7 (d,
2C), 129.7 (d), 131.0 (d, 2C), 146.3 (s, 2C), 149.0 (s, 2C), 160.9
80 (s) ppm; HRMS (ESI+): calcd. For C17H17BrN3O2 (M++H):
374.0499; found 374.0495.
4.1.29.
2-(4-(2-(2,4-Difluorophenyl)-2-hydroxypropyl)-1H-
1,2,3-triazol-1-yl)phenol (3jc). 139 mg (82% yield); Rf = 0.4
(petroleum ether/EtOAc = 7:3); Brown color solid; Mp: 177–179
1
85 °C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.68 (s, 3H),
4.1.24.
2-(4-(2-(3,5-Bis(trifluoromethyl)phenyl)-2-
3.38 (s, 1H), 3.47 (s, 1H), 6.70−6.87 (m, 2H), 6.92−7.10 (m, 2H),
7.23−7.36 (m, 1H), 7.43−7.62 (m, 2H), 7.82 (s, 1H); 13C NMR
[50 MHz, CDCl3 + MeOH (D4)]: δ 28.2 (q), 37.4 (t), 72.7 (s),
103.3 (dd, J = 25.6, 27.4 Hz)), 110.6 (dd, J = 3.3, 23.8 Hz), 117.8
90 (d), 119.9 (d), 122.5 (d), 122.9 (d), 123.7 (s), 128.4 (dd, J =6.4,
9.1 Hz), 129.7 (d), 149.0 (s, 2C), 156.3 (s), 159.2 (s), 161.4 (s)
ppm; HRMS (ESI+): calcd. For C17H16F2N3O2 (M++H):
332.1205; found 332.1199.
25 hydroxyethyl)-1H-1,2,3-triazol-1-yl)-5-chlorophenol (3lb). 119
mg (82% yield); Rf = 0.3 (petroleum ether/EtOAc = 6:4);
Colourless solid; Mp: 192–194 °C; 1H NMR [500 MHz, CDCl3 +
MeOH (D4)]: δ 3.21 (d, J = 6.4 Hz, 2H), 5.19 (t, J = 6.3 Hz, 1H),
6.98 (dd, J = 2.2, 8.6 Hz, 1H), 7.08 (d, J = 2.2 Hz, 1H), 7.60 (d, J
30 = 8.6 Hz, 1H), 7.79 (bs, 2H), 7.88 (bs, 2H), 8.09 (bs, 1H) ppm;
13C NMR [50 MHz, CDCl3 + MeOH (D4)]: δ 34.9 (t), 71.2 (d),
116.7 (d), 119.6 (d, 2C), 120.3 (s), 120.6 (d, t, J = 3.3 Hz), 122.9
(s), 124.8 (d, 2C), 125.8 (d, J = 3.3 Hz), 130.3 (s, d, J = 33.3 Hz ),
131.6 (s, d, J = 32.9 Hz), 134.7 (s, 2C), 146.7 (s, 2C), 149.7 (s)
35 ppm; HRMS (ESI+) calcd for C18H13O2N3ClF6 (M++H):
452.0595; found: 452.0599.
4.1.30.
2-(4-(2-(2,4-Dichlorophenyl)-2-hydroxypropyl)-1H-
95 1,2,3-triazol-1-yl)phenol (3kc). 128 mg (81% yield); Rf = 0.4
(petroleum ether/EtOAc = 7:3);Brown color solid; Mp: 155–157
1
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.71 (s, 3H),
3.39 (d, J = 15.0 Hz, 1H), 3.77 (d, J = 14.9 Hz, 1H), 6.86−7.03
(m, 2H), 7.09−7.25 (m, 2H), 7.28−7.34 (m, 1H), 7.44 (dd, J = 8.0,
100 1.6 Hz, 1H), 7.64 (d, J = 8.6 Hz, 1H), 7.78 (s, 1H); 13C NMR [50
MHz, CDCl3 + MeOH (D4)]: δ 26.8 (q), 35.9 (t), 74.1 (s), 117.5
(d), 119.8 (d), 122.8 (d), 123.1 (d), 126.8 (d), 129.3 (d), 129.6 (d),
130.4 (d), 130.9 (s), 133.0 (s), 142.1 (s), 148.9 (s, 2C), 151.6 (s)
ppm; HRMS (ESI+): calcd. For C17H16Cl2N3O2 (M++H):
105 364.0614; found 364.0611.
4.1.25.
2-(4-(2-(3,5-Bis(trifluoromethyl)phenyl)-2-
hydroxypropyl)-1H-1,2,3-triazol-1-yl)-5-chlorophenol (3mb).
115 mg (80% yield); Colourless solid; Rf = 0.4 (petroleum
40 ether/EtOAc = 6:4); Mp: 156–158 °C; 1H NMR [200 MHz,
CDCl3 + MeOH (D4)]: δ 1.68 (s, 3H), 3.29 (s, 2H), 6.96 (dd, J =
2.2, 8.6 Hz, 1H), 7.07 (d, J = 2.2, 1H), 7.58 (d, J = 8.6 Hz, 1H),
7.76 (bs, 1H), 7.97 (s, 2H), 8.01 (s, 1H) ppm; 13C NMR [50 MHz,
CDCl3 + MeOH (D4)]: δ 28.1 (q), 39.4 (t), 72.8 (s), 116.6 (d),
45 119.4 (d), 119.9 (d, J = 4.0 Hz), 120.3 (s), 122.9 (s), 124.6 (d),
124.8 (d, 2C), 125.1 (dd, J = 2.9 Hz), 125.7 (s), 130.1 (s), 131.4
(s,), 134.6 (s), 142.2 (s), 149.6 (s), 150.2 (s) ppm; HRMS (ESI+)
calcd. for C19H15O2N3ClF6 (M++H): 466.00752; found: 466.0750.
4.1.31. 5-Fluoro-2-(4-(2-(4-fluorophenyl)-2-hydroxypropyl)-
1H-1,2,3-triazol-1-yl)phenol (3gd). 154 mg (83% yield); Rf =
0.3 (petroleum ether/EtOAc = 7:3);Brown color solid; Mp: 186–
1
188 °C; H NMR [500 MHz, CDCl3 + MeOH (D4)]: δ 1.54 (s,
110 3H), 3.21 (s, 2H), 6.87−7.00 (m, 4H), 7.29 (s, 1H), 7.32−7.42 (m,
2H), 7.76 (s, 1H); 13C NMR [125 MHz, CDCl3 + MeOH (D4)]: δ
29.3 (q), 39.9 (t), 73.4 (s), 109.7 (dd, J = 27.7 Hz), 114.6 (dd, J =
21.0 Hz, 2C), 116.1 (dd, J = 22.9 Hz), 118.3 (dd, J = 8.6 Hz),
123.5 (d), 126.5 (dd, J = 8.6 Hz, 2C), 142.8 (s), 144.9 (s), 154.7
115 (s), 156.6 (s), 160.5 (s), 162.5 (s) ppm; HRMS (ESI+): calcd. For
C17H16F2N3O2(M++H): 332.1205; found 332.1199.
4.1.26. 2-(4-(2-(4-Fluorophenyl)-2-hydroxypropyl)-1H-1,2,3-
50 triazol-1-yl)phenol (3gc). 137 mg (78% yield); Rf = 0.3
(petroleum ether/EtOAc = 6:4); Brown color solid; Mp: 156–158
1
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.55 (s, 3H),
3.21 (s, 2H), 6.84−7.05 (m, 4H), 7.14−7.28 (m, 1H), 7.30−7.48
(m, 3H), 7.66 (s, 1H); 13C NMR [50 MHz, CDCl3 + MeOH (D4)]:
55 δ 29.4 (q), 39.9 (t), 73.4 (s), 114.6 (d, J = 21.2 Hz, 2C), 117.8 (d),
119.9 (d), 122.6 (d), 123.1 (d), 123.7 (s), 126.4 (d, J = 7.7 Hz,
2C), 129.7 (d), 142.8 (s), 149.0 (s, 2C), 159.1 (s), 163.9 (s) ppm;
HRMS (ESI+): calcd. For C17H17FN3O2 (M++H): 314.1299;
found 314.1294.
4.1.32. 2-(4-(2-(4-Chlorophenyl)-2-hydroxypropyl)-1H-1,2,3-
triazol-1-yl)-5-fluorophenol (3hd). 135 mg (82% yield); Rf =
0.3 (petroleum ether/EtOAc = 7:3);Light orange color solid; Mp:
1
120 179–181 °C; H NMR [500 MHz, CDCl3 + MeOH (D4)]: δ 1.53
(s, 3H), 3.20 (s, 2H), 6.94 (d, J = 5.8 Hz, 2H), 7.30 (s, 2H), 7.25
60 4.1.27. 2-(4-(2-(4-Chlorophenyl)-2-hydroxypropyl)-1H-1,2,3-
triazol-1-yl)phenol (3hc). 143 mg (84% yield); Rf = 0.3
(s, 1H), 7.31−7.38 (m, 2H), 7.80 (s, 1H); 13C NMR [125 MHz,
8|
Journal Name, [year], [vol], 00–00
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RSC Advances
CDCl3 + MeOH (D4)]: δ 29.6 (q), 39.7 (t), 73.5 (s), 109.2 (m, 60 CLSI broth microdilution assay. For the assay, growth medium
1C), 116.1 (d, J = 22.7 Hz), 118.7 (d, J = 8.2 Hz), 122.9 (d),
123.6 (s), 126.3 (d, 2C), 128.1 (d, 2C), 132.4 (s), 143.6 (s), 145.0
(s), 145.7 (s), 154.8 (s) ppm; HRMS (ESI+): calcd. For
C17H16ClFN3O2 (M++H): 348.0910; found 348.0903.
used was YPG. Briefly, appropriate amount of compounds were
dissolved in dimethyl sulfoxide to get 100X final strength. The
View Article Online
stock was then diluted 1:50 in YPG medDiuOmI: 1a0n.1d03290/0C5μRLA2f5r1o4m5A
this was added to the first row of a 96-well microtitre plate. The
65 compounds were serially diluted two fold in successive wells to
get a range of 1–128 µg/mL. Yeast cells (~2 x 103 cfu/mL),
freshly grown in YPG broth in logarithmic phase, were
suspended in the medium and inoculated (100 μL) in the wells of
the plate. For filamentous fungi, 2 x 104 spores/mL were added.
70 The microtitre plate was incubated for 24 and 48 h for yeasts and
filamentous fungi, respectively. The absorbance was measured at
600 nm by using microtitre plate reader (xMark™ Microplate
Absorbance Spectrophotometer, Bio-Rad, CA, USA) to assess
cell growth. The MIC was defined as the lowest concentration
75 exhibiting >90% inhibition of visible growth as compared to
growth of the control. For determination of MFC, medium from
the MIC and above wells was spot inoculated on 1% YPG agar
plates. The plates were incubated for 48 h, and the minimum
concentration showing no growth was reported as MFC.
5
4.1.33. 2-(4-(2-(4-Bromophenyl)-2-hydroxypropyl)-1H-1,2,3-
triazol-1-yl)-5-fluorophenol (3id). 143 mg (83% yield); Rf = 0.3
(petroleum ether/EtOAc = 7:3); Brown color solid; Mp: 158–160
1
°C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.63 (s, 3H),
10 3.32 (d, J = 1.6 Hz, 2H), 6.97−7.06 (m, 2H), 7.10−7.17 (m,
1H),7.30−7.38 (m, 2H),7.41−7.49 (m, 2H), 7.68 (s, 1H), 9.62 (s,
1H); 13C NMR [50 MHz, CDCl3 + MeOH (D4)]: δ 29.4 (q), 39.6
(t), 73.5 (s), 109.1 (d), 109.6 (d), 115.9 (d), 116.4 (d), 118.5 (dd, J
= 8.8 Hz), 120.5 (s),123.1 (d),126.7 (d, 2C), 131.1 (d, 2C), 143.5
15 (s), 146.2 (s) ppm; HRMS (ESI+): calcd. For C17H16BrFN3O2
(M++H): 392.0404; found 392.0398.
4.1.34.
2-(4-(2-(2,4-Difluorophenyl)-2-hydroxypropyl)-1H-
1,2,3-triazol-1-yl)-5-fluorophenol (3jd). 143 mg (81% yield); Rf
= 0.3 (petroleum ether/EtOAc = 7:3);Brown color solid; Mp:
1
20 174–176 °C; H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 1.66
80 4.4. Haploinsufficiency assay
(s, 3H), 3.33 (d, J = 14.9 Hz, 1H), 3.52 (d, J = 14.9 Hz, 1H),
6.64−6.86 (m, 2H), 6.88−7.09 (m, 2H), 7.14−7.36 (m, 1H),
7.40−7.65 (m, 1H), 7.85 (s, 1H); 13C NMR [50 MHz, CDCl3 +
MeOH (D4)]: δ 28.4 (q), 37.4 (t), 72.8 (s), 104.0 (dd, J = 25.3,
25 27.4 Hz), 108.3 (d), 108.8 (d), 110.8 (dd, J = 3.3, 20.5 Hz), 116.0
(d), 116.4 (d), 119.1 (d, J = 8.42 Hz), 122.2 (d), 128.6 (dd, J =
6.2, 9.5 Hz), 129.4 (s), 143.9 (s), 145.0 (s), 153.4 (s), 158.2 (s)
ppm; HRMS (ESI+): calcd. For C17H15F3N3O2 (M++H):
350.1111; found 350.1104.
In the haploinsufficiency assay, diploid wild type S. cerevisiae
strain and its mutants (including mutants haploid for the genes
erg11 – lanosterol 14 α-demethylase; CHS2 – chitin synthase;
FAB1 - fructose 1,6-bis-phosphate aldolase; FAS2 – Fatty acid
85 synthase; ILV5 – Acedohydroxyacid reductoisomerase etc.) were
used for target identification. The synthesized compounds were
checked for antifungal activity against all the S. cerevisiae strains
in 2X YPG medium by broth microdilution assay (CLSI M27-
A3).13
30 4.1.35.
2-(4-(2-(2,4-Dichlorophenyl)-2-hydroxypropyl)-1H-
1,2,3-triazol-1-yl)-5-fluorophenol (3kd). 144 mg (86% yield);
Rf = 0.4 (petroleum ether/EtOAc = 7:3); Orange color solid; Mp:
156–158 °C; 1H NMR [200 MHz, CDCl3 + MeOH (D4)]: δ 2.3 (s,
3H), 4.03 (d, J = 15.0 Hz, 1H), 4.36 (d, J = 15.0 Hz, 1H),
35 7.50−7.58 (m, 2H), 7.74 (dd, J = 8.6, 2.3 Hz, 1H), 7.88−8.00 (m,
2H), 8.26 (d, J = 8.6 Hz, 1H), 8.6 (s, 1H); 13C NMR [50 MHz,
CDCl3 + MeOH (D4)]: δ 26.6 (q), 35.9 (t), 73.9 (s), 109.7 (d),
110.2 (d), 115.6 (d), 116.1 (d), 117.9 (d), 123.7 (d), 126.6 (d),
129.1 (d), 130.3 (d), 130.9 (s), 132.9 (s), 142.0 (s), 143.2 (s),
40 144.8 (s), 153.1 (s), 157.9 (s) ppm; HRMS (ESI+): calcd. For
C17H15Cl2FN3O2 (M++H): 382.0520; found 382.0515.
90 4.5. Effect of the compounds on sterol profile of C. albicans
ATCC 24433
Depletion of ergosterol as a result of inhibition of lanosterol 14 α-
demethylase by the compounds was quantified by
spectrophotometry.16 Briefly, overnight grown C. albicans ATCC
95 24433 (1 x 106 cfu/mL) cells were inoculated in a series of flasks
with 50 mL of YPG broth containing 8 and 16 μg/mL of the
compounds 3da, 3kb, 3kd, 3la, 3lb, 3ma and 0.125–2 μg/mL
fluconazole. The flasks were incubated on rotary shaker (180
rpm, 30 °C) for 24 h. After incubation, the cells were harvested
100 by centrifugation (3000 rpm for 5 min) and washed with sterile
distilled water. The net wet weight of the cell pellet was
determined. Three mL of 25% alcoholic potassium hydroxide
solution (25 g of KOH and 35 mL of sterile distilled water,
brought to 100 mL with 100% ethanol), was added to 125 mg of
105 pellet and vortex mixed for 1 min. Cell suspensions were
transferred to sterile borosilicate glass screw-cap tubes and were
incubated in an 85 °C water bath for 1 h. Following incubation,
tubes were allowed to cool to room temperature. Sterols were
then extracted by addition of a mixture of 1 mL of sterile distilled
110 water and 3 mL of n-heptane followed by vigorous mixing for 3
min. The heptane layer was scanned between 220 and 300 nm
4.2. Microorganisms and growth conditions
Human pathogens Candida albicans ATCC 24433, Candida
albicans ATCC 10231, Candida glabrata NCYC 388,
45 Cryptococcus neoformans ATCC 34664, Aspergillus fumigatus
NCIM 902, Aspergillus niger ATCC 10578 were maintained on
YPG (yeast extract, 0.3%; peptone, 0.5%; and glucose, 1%) agar
slants at 4 °C. Saccharomyces cerevisiae ATCC 201390 wild
type and various S. cerevisiae haploinsufficient mutants were
50 maintained on 2X YPG agar slants. For different assays, the
inoculum of the yeasts was prepared by growing them in YPG
broth at 28 °C for 24 h.
with
a
spectrophotometer
(Spectrascan
UV-2600
4.3. Antifungal susceptibility assay
Spectrophotometer, Chemito). Ergosterol content was determined
as a percentage of the wet weight of the cells using following
115 equations:
Antifungal activities of the synthesized compounds (in terms of
55 Minimum Inhibitory Concentration; MIC) against C. albicans
ATCC 24433, C. albicans ATCC 10231, C. glabrata NCYC 388,
C. neoformans ATCC 34664, (CLSI - Clinical Laboratory
Standards Institute document M27-A3)13 and A. fumigatus NCIM
902, A. niger ATCC 10578 (CLSI M38-A2)13 were determined by
% ergosterol + % 24(28)DHE = [(A281.5/290) x F]/pellet weight,
% 24(28)DHE = [(A230/518) x F]/pellet weight,
% ergosterol = [% ergosterol + %24(28)DHE] - %24(28)DHE
This journal is © The Royal Society of Chemistry [year]
Journal Name, [year], [vol], 00–00 |9

RSC Advances
Page 10 of 12
Where, F is the dilution factor in ethanol 290 and 518 are the E
G. Medoff, Ann. Rev. Microbiol. 1977, 31, 291; k) H. E. Elphick
values for ergosterol and 24(28)DHE, respectively.
and K. W. Southern, Cochrane Database of Systematic Reviews
2014,
John
Wiley
and View ArtiScloenOsn,line
DOI: 10.1039/C5RA25145A
4.6. Detection of Reactive Oxygen Species (ROS) production
60
DOI:10.1002/14651858.CD002204.pub 3; l) M. V. Castelli, E.
Butassi, M. C. Monteiro, L. A. Svetaz, F. Vicente, and S. A.
Zacchino, Expert Opin. Ther. Pat. 2014, 24, 323.
Intracellular reactive oxygen species (ROS) generation was
assessed by DCFH-DA staining.17 After incubation of the cells of
C. albicans ATCC 24433 with different concentrations of the
compounds for 120 min, 10 μM DCFH-DA in Phosphate Buffer
Saline (PBS, pH 7.4) was added and incubated further for 30 min.
Cells were then harvested, washed with PBS and directly viewed
10 using epifluorescence microscope (LeitzLaborlux S, Germany)
equipped with a 50 W mercury lamp and a filter set (I3 filter
block with excitation filter BP 450–490, and suppression filter
LP520). The digital images were acquired with a Canon
Powershot S80 camera and ZoomBrowser EX 5.5 software for
15 image acquisition and management. The % of fluorescent cells
for each treatment was determined from the number of total
(>400) and fluorescent cells counted.
5
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4.7. Hemolysis assay
All the compounds were tested for RBC hemolysis as described
20 previously.20 The concentrations tested were in the range of 4–
512 μg/mL.
80
Acknowledgements
The antifungal program at CSIR-NCL, Pune, is funded by
Department of Biotechnology, India (grant BT/PR7442/MED/
25 29/680/2012). Y.K. and S.G.T. thank CSIR, India for Research
fellowships.
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*E-mail: sgtupe@yahoo.co.in .
*E-mail: mv.deshpande@ncl.res.in.
*E-mail: vr.chepuri@ncl.res.in
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Graphical Abstract
Re-engineering of PIP3-antagonist triazole PITENIN’s chemical scaffold: development of novel
antifungal leads
Sravani Pulya, Yadagiri Kommagalla, Duhita G. Sant, Shweta U. Jorwekar, Santosh G. Tupe, Mukund
V. Deshpande and Chepuri V. Ramana
CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pune- 411 008, India
designed antifungal agents
anti-cancer agent
Cl
O
Fluconazole
A
B
HO
N
N
Voriconazole
Tebuconazole
Posaconazole
Albaconazole
Ravuconazole
F3C
R
N
OH
Ar
R
HO
Ar
R
N
Ravuconazole
Voriconazole
N
N
N
N
N
N
HO
HO
CF3
Triazole PITENIN
1,2,3-triazoles integrated
with substructural units A and B