Synthesis and Biological Evaluation of Phenyl Substituted 1H-1,2,4-Triazoles as Non-Steroidal Inhibitors of 17β-Hydroxysteroid Dehydrogenase Type 2

Article abstract of DOI:10.1002/ardp.201200025

A series of disubstituted-1H-1,2,4-triazole derivatives was synthesized with the aim of developing new non-steroidal inhibitors of 17β- hydroxysteroid dehydrogenase type 2 (17βHSD2) - a novel and attractive target for the treatment of osteoporosis. 17βHSD2 catalyzes the oxidation of the highly active estrogen 17β-estradiol (E2) and androgen testosterone (T) into the weak estrone and androstenedione, respectively. Inhibition of this enzyme will locally in the bone lead to an increase in E2 and T levels, two key players in the maintenance of the balance between bone resorption and bone formation. In this study, a new class of 17βHSD2 inhibitors with a 1H-1,2,4-triazole scaffold was identified; the three best compounds 8b, 8f, and 13a showed moderate 17βHSD2 inhibitory activity and a good selectivity toward 17βHSD1. They could be a useful tool to map the unexplored enzyme active site. Copyright

Full text of DOI:10.1002/ardp.201200025

Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
1
Full Paper
Synthesis and Biological Evaluation of Phenyl Substituted
1H-1,2,4-Triazoles as Non-Steroidal Inhibitors of
17b-Hydroxysteroid Dehydrogenase Type 2
Yaseen A. Al-Soud1*, Sandrine Marchais-Oberwinkler1, Martin Frotscher1, and Rolf W. Hartmann1,2
1 Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbru¨cken, Saarland, Germany
2 Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research
(HZI), Saarbru¨cken, Saarland, Germany
A series of disubstituted-1H-1,2,4-triazole derivatives was synthesized with the aim of developing
new non-steroidal inhibitors of 17b-hydroxysteroid dehydrogenase type 2 (17bHSD2) – a novel and
attractive target for the treatment of osteoporosis. 17bHSD2 catalyzes the oxidation of the highly
active estrogen 17b-estradiol (E2) and androgen testosterone (T) into the weak estrone and
androstenedione, respectively. Inhibition of this enzyme will locally in the bone lead to an
increase in E2 and T levels, two key players in the maintenance of the balance between bone
resorption and bone formation. In this study, a new class of 17bHSD2 inhibitors with a 1H-1,2,4-
triazole scaffold was identified; the three best compounds 8b, 8f, and 13a showed moderate 17bHSD2
inhibitory activity and a good selectivity toward 17bHSD1. They could be a useful tool to map the
unexplored enzyme active site.
Keywords: 17bHSD2 / Disubstituted 1H-1,2,4-triazoles / Non-steroidal inhibitor / Osteoporosis / Steroidomimetics
Received: January 16, 2012; Revised: March 13, 2012; Accepted: March 15, 2012
DOI 10.1002/ardp.201200025
Introduction
enzymes like aromatase [3–5], CYP17 [6–9], 17bHSD1 [10–18],
5a-reductase [19–23], CYP11B1 [24, 25], and CYP11B2 [26–29].
An intriguing new application for inhibitors interfering
with tissue specific modulation of hormone activity may be
osteoporosis. This skeletal disease is characterized by low
bone mass and deterioration of bone tissue. The disease often
affects elderly women and can be linked to a drop of 17b-
estradiol (E2) levels after menopause. 17b-Hydroxysteroid
dehydrogenase type 2 [30] (17bHSD2) oxidizes E2 into its
inactive form estrone (E1, Chart 1) and is expressed in osteo-
blastic cells which are responsible for bone formation. In
addition, androgens are also known to have beneficial effects
on bone formation [31, 32]. Testosterone, for example, is also
a substrate of 17bHSD2. This enzyme is, therefore, an attrac-
tive target for the treatment of osteoporosis. It has been
validated by Bagi et al. [33] by using an in vivo monkey model.
Not to counteract the therapeutic concept, the type 1 enzyme
catalyzing the reverse reaction (reduction of E1 to E2, Chart 1)
should not be inhibited. Beside the hydroxyphenylnaphthol
A (Chart 2) described by our group [34, 35], there is only one
17b-Hydroxysteroid dehydrogenases (17bHSDs) catalyze the
NAD(P)(H)-dependent interconversion of 17b-hydroxy- and
17-ketosteroids [1]. As both androgens and estrogens display
their maximal biological activity in the reduced (17b-
hydroxy) form, 17bHSDs play a central role in the regulation
of the biological activity of sex steroids. 17bHSD activity is
not limited to steroidogenic tissues, it is also found in a
number of other tissues. The occurrence of activating and
deactivating 17bHSDs is tissue-specific, thus allowing for
local adjustment of hormone action. Selective inhibition
of enzymes involved in the intracellular adjustment of
hormone action is a valuable tool for the treatment of
hormone-sensitive diseases as it has the prospect of less
side-effects compared to systemic hormonal treatments.
This concept was first described for 11bHSDs [2] and has
already been successfully applied by our group for several
Correspondence: Dr. Rolf W. Hartmann, Pharmaceutical and Medicinal
Chemistry, Saarland University, Saarbru¨cken, Saarland, Germany.
E-mail: rwh@mx.uni-saarland.de
Fax: þ49 681 302 70308
ꢀCurrent address: Department of Chemistry, College of Science,
University of Al al-Bayt, Al-Mafraq, Jordan
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Y. A. Al-Soud et al.
Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
OH
OH
O
N
O
S
17β-HSD2, NAD+
N
17β-HSD1, NADPH
HO
HO
1
N
5
B
E2
E1
N
S
2
O
OH
OH
4
N
N
O
3
R2
17β-HSD2, NAD+
R1
O
O
F
1
N
T
5
4
A-dione
N
2
N
R2
3
Chart 1. Interconversion of E2 into estrone (E1) and of testosterone
(T) into androstenedione (A-dione) by 17bHSD2 and 17bHSD1.
N
N
G
OH
N
R1
E
HO
class of non-steroidal 17bHSD2 inhibitors reported in the
literature: the cis-pyrrolidinones like the in vitro highly active
compound B (IC50 ¼ 50 nM in cell-free assay [36], Chart 2).
The latter compound was tested in the monkey model and
had shown moderate effects [33]. This underlines the need for
the discovery of new, more potent 17bHSD2 inhibitors.
In an earlier study [37], different bis(hydroxyphenyl)sub-
stituted heterocycles were evaluated for inhibition of
17bHSD1 and 2. Interestingly, 1H-1,2,3-triazole C (Chart 2)
Chart 3. Designed compounds.
this compound class more thoroughly, trying to improve
17bHSD2 inhibitory activity. As the target enzyme is associ-
ated with the ER membrane there is unfortunately no 3D-
structure available (neither crystal structure nor homology
model has been described to date). Therefore, we followed a
ligand-based drug design approach focusing on the two cis-
pyrrolidinone derivatives B and F (Chart 3) as the triazole
moiety in D and E is a bioisostere of the amide function
present in the pyrrolidinone B. It was then hypothesized that
the thienopyridyl or the diphenyl moiety present in the cis-
pyrrolidinones B and F, respectively, might be important for
inhibitory activity as well as the methyl group and should be
combined and added to the 1H-1,2,4-triazole moiety leading
to derivatives G (Chart 3).
turned out to be
a moderate inhibitor of 17bHSD1
(IC50 ¼ 830 nM), showing good selectivity toward 17bHSD2
while the substituted 1H-1,2,4-triazole D (Chart 2), differing
from C in the presence of a phenyl group on the triazole
moiety and the shift of a nitrogen in the heterocycle, was
active on the type 2 enzyme only (44% inhibition at 1 mM).
The methylated 1H-1,2,4-triazole analogue E was slightly less
active but still selective toward the type 1 enzyme (21%
inhibition at 1 mM). These results caused us to investigate
Based on this hypothesis, in the present study N-methyl-1H-
1,2,4-triazoles bearing an hydroxyphenyl group at C-3 and a
biphenyl or aryl-2-thienyl residue at C-5 were designed, syn-
thesized, and evaluated for their 17bHSD2 and 1 inhibitory
activities (Chart 3).
OH
OH
N
O
S
N
OH
Results and discussion
A
B
Chemistry
Triazole derivatives 8a–h, 8ia–8iq, and 9 bearing substituted
phenyl rings at the C-3 position and substituted biphenyl
residues at the C-5 position were synthesized according to the
route described in Schemes 1 and 2. The 1H-1,2,4-triazole
moiety was prepared in a two steps reaction: first nucleo-
philic substitution of the ethyl imino ester to the acyl
chloride afforded the N-acyliminoester intermediates 1 and
2 (both not isolated). Subsequently nucleophilic addition of
N
N
N
N
N
N
N
OH
OH
N
OH
N
D
C
E
HO
HO
HO
Chart 2. Described inhibitors of 17bHSD2.
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Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
Triazole Derivatives as New Non-Steroidal Inhibitors of 17bHSD2
3
R
O
OEt
O
OEt
N
N
Cl
HN
b
N
N
a
+
X
X
X
R1
R1
R1
Y
Y
Y
3: R=Me, R1= 3-OMe, X=Br, Y=H
4: R=Me, R1= 4-OMe, X=Br, Y=H
5: R=H, R1= 3-OMe, X=Br, Y=H
6: R=H, R1= 4-OMe, X=Br, Y=H
7: R=Me, R1= 3-OMe, X=H, Y=Br
1: R=3-OMe
2: R=4-OMe
B(OH)2
R
R
N
N
N
N
R2
d
N
N
c
R1
HO
R2
OH
8ia: R=Me, R1= 3-OMe, R2=4-OMe
8ib: R=Me, R1= 3-OMe, R2=3-OMe
8ic: R=Me, R1= 4-OMe, R2=4-OMe
8id: R=Me, R1= 4-OMe, R2=3-OMe
8ie: R=H, R1= 3-OMe, R2=4-OMe
8if: R=H, R1= 3-OMe, R2=3-OMe
8ig: R=H, R1= 4-OMe, R2=4-OMe
8ih: R=H, R1= 4-OMe, R2=3-OMe
8a-h
_
Reagents and conditions: a CH2Cl2, NEt3, 30 35 °C, 6 h; b for compounds 3, 4 MeNHNH2 and for compounds 5, 6 NH2NH2.H2O, CH2Cl2,
_
30 40 °C, 4 h; c NaHCO3, Pd(Ph3)4, DMF, microwave irradiation 15 min, 150 °C, 100 W, 15 bar; d BF3.SM2, CH2Cl2, rt, 20 h.
Scheme 1. Synthesis of compounds 8a–h.
hydrazine or methylhydrazine and cyclization led to the
brominated 1H-1,2,4-triazoles 3–7. Suzuki coupling between
the bromo derivatives 3–7 and the appropriate boronic acid
under standard conditions afforded compounds 8ia–8iq.
Ether cleavage using BF3.SMe2 led to the hydroxylated phenyl
derivatives 8a–h and 9 in 44–74 and 68% yield, respectively.
The synthesis of the 1H-1,2,4-triazole arylthiophene deriva-
tives 13a–c was performed following a similar pathway as
described for the biphenyl compounds 8ia–ip in 54–62%
yield (Scheme 3) using the brominated thiophene triazole
11 as key intermediate in the cross-coupling reaction.
The cleavage of the methoxy groups afforded the final
compounds 13a–c using BF3.SMe2.
Biological evaluation and discussion
The synthesized compounds were tested for inhibitory
activity in a cell-free assay using human placental enzymes.
For 17bHSD2 the microsomal fraction and for 17bHSD1
the cytosolic fraction were used as described earlier with
minor modifications [38]. Results are shown in Table 1 as
percentage inhibition measured at a concentration of 1 mM.
Compounds showing less than 10% inhibition at 1 mM were
considered to be inactive. For the sake of clarity in this
manuscript and to be able to distinguish the two hydroxy-
phenyl moities, they were named A- and B-ring, respectively
(Table 1).
The most active compounds 8b, 8f, and 13a show inhi-
bition of the target enzyme of 42, 20, and 30%, respectively
(Table 1). It is interesting to notice that all these compounds
bear one hydroxy group on each phenyl ring (A and B) while
the corresponding methoxy derivatives are all inactive
1
The physical characteristics observed in the H NMR and
13C NMR spectra (including DEPT experiments) and LC-MS
spectra of all new prepared compounds are in total agree-
ment with the suggested structures.
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Y. A. Al-Soud et al.
Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
B(OH)2
Me
R
N
N
N
N
N
N
R2
7
b
a
R2
OMe
OH
R3
OH
R4
9
8ii : R2=H, R3=OMe, R4=H
8ij : R2=H, R3=OH, R4=H
8ik : R2=F, R3=OMe, R4=H
8il: R2=H, R3=F, R4=OMe
8im : R2=H, R3=NH2, R4=H
8in : R2=H, R3=H, R4=OH
8io : R2=H, R3=H, R4=CN
8ip : R2=Me, R3=H, R4=F
8iq : R2=H, R3=F, R4=H
Reagents and conditions: a NaHCO3, Pd(Ph3)4, DMF, microwave irradiation 15 min, 150 °C,
100 W, 15 bar;b BF3.SM2, CH2Cl2, rt, 20 h.
Scheme 2. Synthesis of compounds 8ii–iq and 9.
(results not shown). This characteristic has already been
observed in the development of 17bHSD2 [10, 34] as well
as 17bHSD1 inhibitors [10]. In addition, it is striking that all
the active compounds 8b, 8f, and 13a have both OH groups in
meta-position of the respective aromatic rings. Compound 8d,
with a meta,para-substitution pattern for both OH groups
shows reduced inhibitory activity compared to the meta,meta
8b although the distance between the two OH groups does
Me
OEt
O
OEt
N
N
O
Br
b
HN
N
S
N
a
+
S
S
Cl
Br
Br
OMe
OMe
OMe
11
10
B(OH)2
Me
Me
N
N
N
N
X
R2
d
N
N
S
S
c
X
X
OH
OMe
OH
R
12a: X=CH, R=3-OMe
12b: X=CH, R=4-OMe
12c: X=N, R=4-OMe
_
13a c
_
_
Reagents and conditions: a CH2Cl2, NEt3, 30 40 °C, 6 h;b MeNHNH2, CH2Cl2, 30 40 °C, 4 h;c
NaHCO3, Pd(Ph3)4, DMF, microwave
irradiation 15 min, 150 °C, 100 W, 15 bar; d BF3.SM2,CH2Cl2, rt, 20 h.
Scheme 3. Synthesis of compounds 13a–c.
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Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
Triazole Derivatives as New Non-Steroidal Inhibitors of 17bHSD2
5
Table 1. Human 17bHSD2 and 17bHSD1 inhibition activity of compounds 8a–h, 8ii–iq, 9, and 13a–c
H
3C
R
R
N
N
N N
N
N
N
N
N
B
B
B
S
R1
X
R1
A
A
R
R3
R2
R2
A
_
_
_
a c
a
8
13
h
8ii i
9
,
q
R1
R2
Compounds
R
R1
R2
R3
X
17bHSD2a) % Inhibition
at 1 mM
17bHSD1b) % Inhibition
at 1 mM
8a
8b
8c
8d
8e
8f
8g
8h
8ii
8ij
8ik
8il
8im
8in
8io
8ip
8iq
9
CH3
CH3
CH3
CH3
H
H
H
H
H
H
F
H
H
H
H
CH3
H
H
3-OH
3-OH
4-OH
4-OH
3-OH
3-OH
4-OH
4-OH
OCH3
OH
OCH3
F
NH2
H
H
H
4-OH
3-OH
4-OH
3-OH
4-OH
3-OH
4-OH
3-OH
H
ni
ni
12
ni
14
ni
23
ni
27
ni
ni
ni
ni
ni
ni
ni
ni
ni
ni
ni
ni
ni
42%, IC50 ¼ 1.4 mMc)
ni
14
ni
20%, IC50 ¼ 4.0 mMc)
ni
ni
ni
ni
ni
ni
ni
ni
ni
ni
ni
ni
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OH
H
H
OCH3
H
OH
CN
F
H
H
3-OH
4-OH
4-OH
F
OH
3-OH
3-OH
3-OH
13a
13b
13c
CH3
CH3
CH3
CH
CH
N
30%, IC50 ¼ 2.0 mMc)
ni
ni
Human placenta, microsomal fraction, substrate [3H]-E2 þ E2 [500 nM], cofactor NADþ [1500 mM].
Human placenta, cytosolic fraction, substrate [3H]-E1 þ E1 [500 nM], cofactor NADH [500 mM].
a)
b)
c)
IC50-value (calculated from % inhibition via logit transformation); ni, no inhibition (<10% inhibition at 1 mM).
˚
not vary so much compared to 13a (14.00 A in 13a and
˚
14.81 A in 8d). From this it has to be concluded that
ring and the triazole moieties fixed). The most favorable
conformation has a linear shape as facilitated by the 1,4-
phenyl group.
H-bonding interactions cannot be the main stabilizing force
for the compounds, electronic effects might be involved as
well as Van der Waals interactions.
It can also be noticed that the N-methyl triazole 8b
has a slightly higher inhibitory activity compared to 8f
without methyl group. This hydrophobic group interacts
with a small lipophilic pocket, as hypothesized with the
cis-pyrrolidinone B.
Replacement of the central phenyl ring (8b and 8f) by a
thiophene (13a) seems to have a negligible influence on the
biological activity, indicating that the S of the thiophene
does not achieve any specific interaction. It should also
be noticed that 17bHSD2 inhibitory activity varies with
the substitution pattern of the central ring (1,4-phenyl in
8b being the most active, 2,5-thiophene in 13a and 1,3-phenyl
in 8f weaker). This difference in activity is certainly
linked to the position of the OH-phenyl group which covers
a different area of the active site in each case (having the B
In addition, the overall large size of 8b and 13a indicates
that the active site of the enzyme must be quite elongated
and able to accept big compounds.
Compounds 8ii–8iq with 1,3-phenyl as central ring with a
U-shape were prepared to mimic the conformation of the cis-
pyrrolidinone B. Their inactivity might indicate that either
these compounds do not bind in the same way as B or that the
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Y. A. Al-Soud et al.
Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
flatness and the rigidity of the 1H-1,2,4-triazole is detrimental
for activity.
General procedure for the synthesis of compounds 5, 6,
and 11 – formation of the 1H-1,2,4-triazole moiety
A solution of acyl chloride (1 equiv) in 10 mL dichloromethane
was added dropwise to a mixture of ethyl imino ester (1 equiv)
and anhydrous triethylamine (1 equiv) in 20 mL dichloro-
methane at rt. The reaction mixture was heated to 30–358C for
6 h. After cooling to rt, the material was poured into a 3% sodium
hydrogenocarbonate solution (25 mL). The layers were separated
and the organic layer was washed with water, dried over sodium
sulfate, and evaporated to dryness under reduced pressure. The
resulting N-acylimino esters 1, 2, and 10 were not characterized.
They were used in the next step without further purification. They
were heated to 30–358C with the hydrazine derivative (2 equiv) in
dichloromethane for 4 h. The solvent was removed under reduced
pressure. The 1H-1,2,4-triazoles were purified by recrystallization
using CH2Cl2/diethyl ether for compounds 3, 4, and 11 and EtOH/
petroleum ether for compounds 5 and 6.
Comparing the 1H-1,2,3-triazole C (inhibitor of 17bHSD1)
with the 1H-1,2,4-triazole E or 8b (17bHSD2 inhibitor), it is
striking that the change in position of one N only within the
triazole moiety allows a switch in selectivity. It demonstrates
the high differences in both enzyme binding sites.
The 17bHSD2 inhibitory activity of the newly synthesized
compound is moderate (40% inhibition at 1 mM for 8b) but a
rather good selectivity is achieved toward 17bHSD1 (12%
inhibition at 1 mM for 8b).
Conclusion
In this study, a series of 21 1H-1,2,4-triazole derivatives was
synthesized and tested for 17bHSD2 and 17bHSD1 inhi-
bition. Three compounds 8b, 8f, and 13a were identified as
moderate 17bHSD2 inhibitors, 8b being the best identified
with an IC50 of 1.4 mM. They are selective toward 17bHSD1.
They all share an N-methyl 1H-1,2,4-triazole ring as central
core and two meta,meta-hydroxyphenyl substituents. These
compounds need to be further optimized but they can
already be considered as a good tool for the development
of further 17bHSD2 inhibitors and for the mapping of the
17bHSD2 active site.
5-(4-Bromophenyl)-3-(3-methoxyphenyl)-1H-1,2,4-
triazole 5
The title compound was prepared from 4-bromobenzoyl chloride
(0.219 g, 1.0 mmol), ethyl 3-methoxybenzimidate (0.179 g,
1.0 mmol), and hydrazine hydrate (0.100 g, 2.0 mmol) according
to the procedure described above. The product was crystallized
from EtOH (5 mL)/Petroleum ether (1 mL). Yield: 0.180 g (54%).
C15H12BrN3O; MW: 330; m.p. 191–1938C (dec). 1H NMR (DMSO-d6):
d 9.85 (bs, 1H, NH); 8.02 (d, J ¼ 8.5 Hz, 2H); 7.71 (d, J ¼ 8.5 Hz,
2H); 7.67–7.62 (m, 2H); 7.44 (t, J ¼ 7.9 Hz, 1H); 7.05 (m, 1H); 3.84
(s, 3H, OCH3). 13C NMR (DMSO-d6): d 159.6, 157.4, 131.8, 131.3,
130.1, 129.7, 129.0, 127.9, 122.8, 118.4, 115.7, 111.1, 55.2 (OCH3).
MS (ESI): 329–331 [MþH]þ.
Experimental section
5-(4-Bromophenyl)-3-(4-methoxyphenyl)-1H-1,2,4-
triazole 6
General
Chemical names follow IUPAC nomenclature.
Starting materials were purchased from Aldrich, Acros,
Lancaster, Roth, Merck, or Fluka and were used without
purification.
Purification was performed on preparative thin layer chroma-
tography (TLC) on 1 mm SIL G-100 UV254 glass plates (Macherey-
Nagel, Du¨ren) and reaction progress was monitored by TLC on
Alugram SIL G UV254 (Macherey-Nagel). Visualization was
accomplished with UV light.
Melting points were measured on a Mettler FP1 melting point
apparatus and are uncorrected. 1H NMR and 13C NMR spectra
were measured on a Bruker AM500 spectrometer (500 MHz) at
300 K. Chemical shifts are reported in d (parts per million: ppm),
by reference to the hydrogenated residues of deuteriated solvent
as internal standard (CDCl3: d 7.24 ppm (1H NMR) and d 77 ppm
The title compound was prepared from 4-bromobenzoyl chloride
(0.219 g, 1.0 mmol), ethyl 4-methoxybenzimidate (0.179 g,
1.0 mmol), and hydrazine hydrate (0.100 g, 2.0 mmol) according
to the procedure described above. The product was crystallized
from EtOH (5 mL)/Petroleum ether (1 mL). Yield: 0.175 g (53%).
1
C15H12BrN3O; MW: 330; m.p. 223–2258C (dec). H NMR (DMSO-d6):
d 14.4 (bs, NH); 8.02 (d, J ¼ 3.9 Hz, 2H); 8.00 (d, J ¼ 4.5 Hz, 2H); 7.70
(d, J ¼ 8.1 Hz, 2H); 7.09 (d, J ¼ 8.5 Hz, 2H); 3.82 (s, 3H, OCH3).
13C NMR (DMSO-d6): d 160.6, 160.5, 131.8, 131.3, 129.0, 127.9,
127.6, 122.5, 114.3, 55.3 (OCH3). MS (ESI): 329–331 [MþH]þ.
5-(5-Bromo-2-thienyl)-3-(3-methoxyphenyl)-1-methyl-1H-
1,2,4-triazole 11
The title compound was prepared from 5-bromothiophene-2-
carbonyl chloride (0.225 g, 1.0 mmol), ethyl 3-methoxybenzimi-
date (0.179 g, 1.0 mmol), and methyl hydrazine (0.092 g,
2.0 mmol) according to the procedure described above. The
product was crystallized from CH2Cl2 (5 mL)/Et2O (1 mL). Yield:
0.302 g (86%). C14H12BrN3OS; MW: 350; m.p. 106–1088C (dec).
1H NMR (CDCl3): d 7.71–7.69 (m, 1H); 7.65–7.64 (m, 1H); 7.34
(t, J ¼ 7.9 Hz, 1H); 7.26 (d, J ¼ 3.9 Hz, 1H); 7.13 (d, J ¼ 4.0 Hz,
1H); 6.96–6.94 (m, 1H); 4.06 (s, 3H, NCH3); 3.88 (s, 3H, OCH3).
13C NMR (CDCl3): d 161.0, 159.8, 149.0, 131.8, 131.0, 130. 7, 129.6,
128.2, 118.8, 116.2, 115.8, 111.0, 55.4 (OCH3), 37.1 (NCH3).
MS (ESI): 349–351 [MþH]þ.
(
13C NMR); CD3OD: d 3.35 ppm (1H NMR) and d 49.3 ppm (13
C
NMR); DMSO-d6: d 2.50 ppm (1H NMR), d 39.5 ppm (13C NMR).
Signals are described as s, d, t, dd, and m, for singlet, doublet,
triplet, doublet of doublets, and multiplet, respectively. All
coupling constants (J) are given in Hertz (Hz).
Mass spectra (ESI) were recorded on a TSQ Quantum (Thermo
Fisher) instrument using electrospray (ESI) as ionization source.
All microwave irradiation experiments were carried out in a
CEM-Discover microwave apparatus.
Compounds 3 [39], 4 [39], and 7 [40] were prepared according to
the previously described procedure.
ß 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
Triazole Derivatives as New Non-Steroidal Inhibitors of 17bHSD2
7
General procedure for compounds 8ia–ih, 8ii–iq and
12a–c – Suzuki coupling
5-(40-Methoxybiphenyl-4-yl)-3-(4-methoxyphenyl)-1-
methyl-1H-1,2,4-triazole 8ic
Method A: Boronic acid (0.75 mmol, 1 equiv), 1H-1,2,4-triazolyl-
bromide (1 equiv), and tetrakis(triphenylphosphane)palla-
dium(0) (43 mg, 0.0375 mmol, 5 mol %) were suspended in
1.5 mL DMF in a 10 mL septum-capped tube containing a stirring
magnet. A solution of NaHCO3 (189 mg, 2.25 mmol, 3 equiv)
in 1.5 mL water was added and the vial was sealed with a
Teflon cap. The mixture was irradiated with microwaves
for 15 min at a temperature of 1508C with an initial irradiation
power of 100 W at 15 bar. After the reaction, the vial was
cooled to rt, the crude mixture was partitioned between ethyl
acetate and water, and the aqueous layer was extracted
three times with ethyl acetate. The combined organic layers
were dried over MgSO4 and the solvents were removed in vacuo.
The coupling products were obtained after purification by
preparative TLC.
Method B: A mixture of arylbromide (1 equiv), boronic
acid derivative (1.2 equiv), cesium carbonat (4 equiv), and tetra-
kis(triphenylphosphine) palladium (0.03 equiv) in an oxygen-
free DME/water (1:1) mixture was stirred at 1508C for 6 h under
nitrogen atmosphere. The reaction mixture was cooled to rt. The
aqueous layer was extracted with ethyl acetate. The combined
organic layers were washed with brine, dried over sodium
sulfate, filtered, and concentrated to dryness. The product was
purified by preparative TLC.
The title compound was prepared by reaction of 5-(4-bromophenyl)-3-
(4-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (4) (0.344 g, 1.0 mmol)
with 4-methoxybenzene boronic acid (0.114 g, 0.75 mmol) accord-
ing to method A. The product was purified by preparative TLC
(CH2Cl2/MeOH, 10:0.25); yield: 0.200 g (54%). C23H21N3O2; MW:
1
371; m.p. 137–1398C (dec), white powder. H NMR (CDCl3): d 8.07
(d, J ¼ 8.8 Hz, 2H); 7.76 (d, J ¼ 8.0 Hz, 2H); 7.68 (d, J ¼ 8.3 Hz, 2H);
7.57 (d, J ¼ 8.6 Hz, 2H); 6.99 (d, J ¼ 8.6 Hz, 2H); 6.95 (d, J ¼ 8.8 Hz,
2H); 4.02 (s, 3H, NCH3); 3.85 (s, 3H, OCH3); 3.84 (s, 3H, OCH3).
13C NMR (CDCl3): d 161.2, 160.7, 160.0, 155.5, 142.7, 132.7, 130.3,
129.4, 128.5, 128.0, 127.2, 124.0, 114.7, 114.2, 55.6 (OCH3), 55.5
(OCH3), 37.2 (NCH3). MS (ESI): 372 [MþH]þ.
5-(30-Methoxybiphenyl-4-yl)-3-(4-methoxyphenyl)-1-
methyl-1H-1,2,4-triazole 8id
The title compound was prepared by reaction of 5-(4-bromo-
phenyl)-3-(4-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (4)
(0.344 g, 1.0 mmol) with 3-methoxybenzene boronic acid
(0.114 g, 0.75 mmol) according to method A. The product was
purified by preparative TLC (CH2Cl2/MeOH, 10:0.25); yield:
0.220 g (59%). C23H21N3O2; MW: 371; m.p. 154–1568C (dec), white
powder. 1H NMR (CDCl3): d 8.09 (d, J ¼ 8.8 Hz, 2H); 7.80
(d, J ¼ 8.3 Hz, 2H); 7.74 (d, J ¼ 8.3 Hz, 2H); 7.40 (t, J ¼ 8.0 Hz,
1H); 7.24–7.17 (m, 2H); 6.97 (d, J ¼ 8.8 Hz, 2H); 6.95 (m, 1H); 4.05
(s, 3H, NCH3); 3.89 (s, 3H, OCH3); 3.86 (s, 3H, OCH3). 13C NMR
(CDCl3): d 161.4, 160.7, 160.33, 155.5, 142.9, 141.8, 130.2, 129.4,
128.0, 127.8, 127.3, 124.1, 119.9, 114.2, 113.5, 113.3, 55.6 (OCH3),
55.5 (OCH3), 37.2 (NCH3). MS (ESI): 372 [MþH]þ.
5-(40-Methoxybiphenyl-4-yl)-3-(3-methoxyphenyl)-1-
methyl-1H-1,2,4-triazole 8ia
The title compound was prepared by reaction of 5-(4-bromo-
phenyl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole
(3)
5-(40-Methoxybiphenyl-4-yl)-3-(3-methoxyphenyl)-1H-
1,2,4-triazole 8ie
(0.344 g, 1.0 mmol) with 4-methoxybenzene boronic acid
(0.114 g, 0.75 mmol) according to method A. The product was
purified by preparative TLC (CH2Cl2/MeOH, 10:0.25); yield:
0.227 g (61%). C23H21N3O2; MW: 371; m.p. 147–1498C (dec), white
powder; 1H NMR (DMSO-d6): d 7.90 (d, J ¼ 6.8 Hz, 2H); 7.83
(d, J ¼ 8.4 Hz, 2H); 7.72 (d, J ¼ 8.8 Hz, 2H); 7.65–7.64 (m, 1H);
7.41–7.38 (m, 1H); 7.40 (t, J ¼ 7.9 Hz, 1H); 7.07 (t, J ¼ 8.0 Hz, 2H);
7.02–7.00 (m, 1H); 4.05 (s, 3H, NCH3); 3.83 (s, 3H, OCH3); 3.81
(s, 3H, OCH3). 13C NMR (DMSO-d6): d 159.5, 159.5, 159.4, 154.6,
141.2, 132.3, 131.3, 129.9, 129.1, 127.9, 126.4, 125.8, 118.1, 115.0,
114.5, 110.6, 55.2 (OCH3), 55.1 (OCH3), 37.2 (NCH3); MS (ESI): 372
[MþH]þ.
The title compound was prepared by reaction of 5-(4-bromo-
phenyl)-3-(3-methoxyphenyl)-1H-1,2,4-triazole
(5)
(0.330 g,
1.0 mmol) with 4-methoxybenzene boronic acid (0.114 g,
0.75 mmol) according to method A. The product was purified
by preparative TLC (CH2Cl2/MeOH, 10:1); yield: 0.158 g (44%).
C22H19N3O2; MW: 357; m.p. 178–1808C (dec), white powder.
1H NMR (CD3OD þ CDCl3; 2:1): d 8.08 (d, J ¼ 8.2 Hz, 2H); 7.68
(d, J ¼ 8.2 Hz, 2H); 7.64 (d, J ¼ 6.7 Hz, 2H); 7.58 (d, J ¼ 8.7 Hz,
2H); 7.50–7.45 (m, 1H); 7.39 (t, J ¼ 8.1 Hz, 1H); 7.00–6.98
(m, 2H); 3.88 (s, 3H, OCH3); 3.83 (s, 3H, OCH3). 13C NMR
(CD3OD þ CDCl3; 2:1):
d 160.1, 159.7, 142.3, 132.5, 129.8,
129.7, 128.6, 127.8, 126.7, 126.6, 126.3, 118.7, 118.6, 115.8,
5-(30-Methoxybiphenyl-4-yl)-3-(3-methoxyphenyl)-1-
methyl-1H-1,2,4-triazole 8ib
114.1, 111.4, 54.7 (OCH3), 54.6 (OCH3). MS (ESI): 358 [MþH]þ.
The title compound was prepared by reaction of 5-(4-bromo-
phenyl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole
5-(30-Methoxybiphenyl-4-yl)-3-(3-methoxyphenyl)-1H-
1,2,4-triazole 8if
(3)
(0.344 g, 1.0 mmol) with 3-methoxybenzene boronic acid
(0.114 g, 0.75 mmol) according to method A. The product
was purified by preparative TLC (CH2Cl2/MeOH, 10:0.25);
yield: 0.215 g (58%). C23H21N3O2; MW: 371; m.p. 123–1258C
(dec), white powder. 1H NMR (DMSO-d6): d 7.93 (d, J ¼ 8.5 Hz,
2H); 7.88 (d, J ¼ 8.5 Hz, 2H); 7.66–7.65 (m, 1H); 7.59–7.57 (m, 1H);
7.44–7.39 (m, 2H); 7.34–7.29 (m, 2H); 7.02–6.99 (m, 2H); 4.06
(s, 3H, NCH3); 3.85 (s, 3H, OCH3); 3.83 (s, 3H, OCH3). 13C NMR
(DMSO-d6): d 159.8, 159.5, 154.5, 141.5, 140.6, 132.2, 130.1,
130.1, 129.1, 127.6, 127.1, 126.7, 119.1, 118.1, 115.0, 113.7,
112.3, 110.6, 55.2 (OCH3), 55.1 (OCH3), 37.2 (NCH3). MS (ESI):
372 [MþH]þ.
The title compound was prepared by reaction of 5-(4-bromo-
phenyl)-3-(3-methoxyphenyl)-1H-1,2,4-triazole
(5)
(0.330 g,
1.0 mmol) with 3-methoxybenzene boronic acid (0.114 g,
0.75 mmol) according to method A. The product was purified
by preparative TLC (CH2Cl2/MeOH, 10:1); yield: 0.211 g
(59%). C22H19N3O2; MW: 357; m.p. 108–1108C (dec), white pow-
der. 1H NMR (CD3OD þ CDCl3; 2:1): d 8.10 (d, J ¼ 8.2 Hz, 2H); 7.69
(d, J ¼ 8.3 Hz, 2H); 7.64–7.60 (m, 2H); 7.46–7.36 (m, 1H); 7.31
(t, J ¼ 7.9 Hz, 1H); 7.19–7.14 (m, 2H); 6.99–6.89 (m, 2H); 3.85
(s, 3H, OCH3); 3.81 (s, 3H, OCH3). 13C NMR (CD3OD þ CDCl3;
2:1): d 161.5, 161.5, 143.9, 143.0, 131.4, 131.2, 131.1, 130.1,
ß 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

8
Y. A. Al-Soud et al.
Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
129.2, 128.7, 128.2, 127.8, 120.7, 120.1, 117.3, 114.4, 113.9, 112.9,
56.3 (OCH3), 56.2 (OCH3). MS (ESI): 358 [MþH]þ.
129.0, 120.3, 120.2, 116.8, 116.6, 115.8, 113.0, 55.4 (OCH3), 55.4
(OCH3), 37.0 (NCH3). MS (ESI): 358 [MþH]þ.
5-(40-Methoxybiphenyl-4-yl)-3-(4-methoxyphenyl)-1H-
1,2,4-triazole 8ig
5-(20-Fluoro-30-methoxybiphenyl-3-yl)-3-
(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole 8ik
The title compound was prepared by reaction of 5-(4-bromophenyl)-
3-(4-methoxyphenyl)-1H-1,2,4-triazole (6) (0.330 g, 1.0 mmol) with
4-methoxybenzene boronic acid (0.114 g, 0.75 mmol) according to
method A. The product was purified by preparative TLC (CH2Cl2/
MeOH, 10:1); yield: 0.185 g (52%). C22H19N3O2; MW: 357; m.p. 202–
2048C (dec), white powder. 1H NMR (CD3OD): d 8.01 (d, J ¼ 7.9 Hz,
2H); 8.00 (d, J ¼ 8.3 Hz, 2H); 7.72 (d, J ¼ 8.0 Hz, 2H); 7.63 (d, J ¼
8.4 Hz, 2H); 7.07 (d, J ¼ 8.4 Hz, 2H); 7.02 (d, J ¼ 8.6 Hz, 2H); 3.87
(s, 3H, OCH3); 3.84 (s, 3H, OCH3). 13C NMR (CD3OD): d 161.2, 161.1,
133.8, 133.1, 133.1, 130.0, 129.9, 129.2, 129.1, 128.0, 127.9, 127.6,
118.7, 115.4, 55.9 (OCH3), 55.8 (OCH3). MS (ESI): 358 [MþH]þ.
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
2-fluoro-3-methoxyphenylboronic acid (0.204 g, 1.2 mmol) accord-
ing to method B. The product was purified by preparative TLC
(CH2Cl2); yield: 0.266 g (68%). C22H19N3O2; MW: 389; m.p. 119–
1218C (dec); white powder. 1H NMR (CDCl3): 7.80–7.79 (m, 1H);
7.66–7.64 (m, 2H); 7.60–7.56 (m, 2H); 7.94 (t, J ¼ 7.6 Hz, 1H,);
7.75–7.70 (m, 4H); 7.62 (t, J ¼ 7.8 Hz, 1H); 7.24 (t, J ¼ 7.9 Hz,
1H); 7.06–7.03 (m, 1H); 6.96–6.83 (m, 3H); 3.95 (s, 3H, NCH3); 3.83
(s, 3H, OCH3); 3.78 (s, 3H, OCH3). 13C NMR (CDCl3): d 161.1, 159.9,
155.4, 150.6, 148.6, 148.3, 136.3, 132.3, 130.7, 129.6, 129.4, 129.1,
128.1, 124.2, 124.1, 121.9, 118.8, 115.8, 112.8, 110.8, 56.4 (OCH3),
55.4 (OCH3), 37.0 (NCH3). MS (ESI): 390 [MþH]þ.
5-(30-Methoxybiphenyl-4-yl)-3-(4-methoxyphenyl)-1H-
1,2,4-triazole 8ih
5-(30-Fluoro-40-methoxybiphenyl-3-yl)-3-
The title compound was prepared by reaction of 5-(4-bromophenyl)-
3-(4-methoxyphenyl)-1H-1,2,4-triazole (6) (0.330 g, 1.0 mmol) with
3-methoxybenzene boronic acid (0.114 g, 0.75 mmol) according to
method A. The product was purified by preparative TLC (CH2Cl2/
MeOH, 10:1); yield: 0.171 g (48%). C22H19N3O2; MW: 357; m.p. 131–
1338C (dec), white powder. 1H NMR (CD3OD þ CDCl3; 2:1): d 8.08–
8.02 (m, 2H); 7.96–7.91 (m, 2H); 7.65 (d, J ¼ 8.3 Hz, 2H); 7.45–7.41
(m, 1H); 7.29 (t, J ¼ 7.9 Hz, 1H); 7.17–7.12 (m, 1H); 6.97
(d, J ¼ 8.6 Hz, 2H); 6.87–6.85 (m, 1H); 3.79 (s, 3H, OCH3); 3.78
(s, 3H, OCH3). 13C NMR (CD3OD): d 162.7, 161.5, 143.7, 143.0,
131.1, 130.0, 129.3, 128.5, 128.1, 127.7, 122.4, 122.3, 120.6, 115.5,
114.3, 113.8, 56.1 (OCH3), 56.0 (OCH3). MS (ESI): 358 [MþH]þ.
(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole 8il
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
3-fluoro-4-methoxyphenylboronic acid (0.204 g, 1.2 mmol) accord-
ing to method B. The product was purified by preparative TLC
(CH2Cl2); yield: 0.278 g (71%). C22H19N3O2; MW: 389; m.p. 99–
1018C (dec); yellow powder. 1H NMR (CDCl3): 8.04 (bs, 1H); 7.85
(d, J ¼ 7.7 Hz, 1H); 7.78 (d, J ¼ 7.7 Hz, 1H); 7.69–7.56 (m, 5H); 7.40
(t, J ¼ 7.9 Hz, 1H); 7.27 (t, J ¼ 7.7 Hz, 1H); 7.00 (d, J ¼ 8.2 Hz, 1H,);
4.05 (s, 3H, NCH3); 3.90 (s, 3H, OCH3); 3.83 (s, 3H, OCH3). 13C NMR
(CDCl3): d 159.5, 159.4, 154.7, 152.7, 150.8, 146.9, 139.2, 132.2,
129.8, 129.4, 128.3, 127.9, 127.4, 126.3, 123.0, 118.1, 114.9, 114.4,
114.2, 110.6, 56.4 (OCH3), 55.4 (OCH3), 37.0 (NCH3). MS (ESI): 390
[MþH]þ.
5-(30-Methoxybiphenyl-3-yl)-3-(3-methoxyphenyl)-1-
methyl-1H-1,2,4-triazole 8ii
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
3-methoxybenzene boronic acid (0.182 g, 1.2 mmol) according to
method B. The product was purified by preparative TLC (CH2Cl2);
30-[3-(3-Methoxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
biphenyl-3-amine 8im
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
3-aminophenylboronic acid (0.164 g, 1.2 mmol) according to
method B. The product was purified by preparative TLC (CH2Cl2);
yield: 0.258 g (72%). C22H20N4; MW: 356; m.p. 142–1448C (dec);
yellow powder. 1H NMR (CD3COCD3): 8.05 (t, J ¼ 1.7 Hz, 1H);
7.81–7.79 (m, 1H); 7.78–7.74 (m, 2H); 7.72–7.71 (m, 1H); 7.63
(t, J ¼ 7.9 Hz, 1H); 7.38 (t, J ¼ 7.9 Hz, 1H); 7.18 (t, J ¼ 7.8 Hz,
1H); 7.06 (t, J ¼ 2.0 Hz, 1H); 6.99–6.95 (m, 2H); 6.73–6.71 (m, 1H);
4.76 (bs, 2H, NH2); 4.11 (s, 3H, NCH3); 3.87 (s, 3H, OCH3). 13C NMR
(CD3COCD3): d 162.2, 161.9, 157.2, 151.0, 144.3, 142.8, 134.9, 131.5,
131.5, 131.0, 130.8, 130.1, 129.1, 129.0, 120.3, 117.4, 116.6, 115.8,
114.8, 113.0, 56.6 (OCH3), 38.7 (NCH3). MS (ESI): 357 [MþH]þ.
1
yield: 0.250 g (67%). C23H21N3O2; MW: 371; oily product. H NMR
(CDCl3): d 7.95 (s, 1H); 7.77 (d, J ¼ 7.7 Hz, 1H,); 7.73–7.63 (m, 3H);
7.59 (t, J ¼ 7.8 Hz, 1H); 7.40–7.34 (m, 2H); 7.23–7.17 (m, 2); 6.97–6.94
(m, 2H); 4.05 (s, 3H, NCH3); 3.89 (s, 3H, OCH3); 3.88 (s, 3H, OCH3).
13C NMR (CDCl3): d 161.2, 160.1, 159.9, 155.6, 142.0, 141.7, 132.4,
130.0, 129.7, 129.3, 129.0, 128.6, 127.7, 127.6, 119.8, 118.9, 115.9,
113.2, 113.1, 110.9, 55.4 (OCH3), 55.4 (OCH3), 37.0 (NCH3). MS (ESI):
372 [MþH]þ.
30-[3-(3-Methoxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
biphenyl-3-ol 8ij
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
3-hydroxybenzene boronic acid (0.166 g, 1.2 mmol) according to
method B. The product was purified by preparative TLC (CH2Cl2);
yield: 0.284 g (79%). C22H19N3O2; MW: 357; m.p. 180–1828C (dec);
white powder. 1H NMR (CD3COCD3): 8.50 (bs, 1H, OH); 8.08
(t, J ¼ 1.7 Hz, 1H); 7.85–7.79 (m, 2H); 7.75–7.71 (m, 2H); 7.66
(t, J ¼ 7.8 Hz, 1H); 7.38 (t, J ¼ 7.8 Hz, 1H); 7.33 (t, J ¼ 8.0 Hz, 1H);
7.23–7.21 (m, 2H); 6.99–6.97 (m, 1H); 6.91–6.88 (m, 1H); 4.13 (s, 3H,
NCH3); 3.87 (s, 3H, OCH3). 13C NMR (CD3COCD3): d 162.2, 162.0, 159.9,
157.1, 143.5, 143.5, 134.9, 132.0, 131.5, 131.2, 130.9, 130.2, 129.5,
30-[3-(3-Methoxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
biphenyl-4-ol 8in
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
4-hydroxybenzene boronic acid (0.166 g, 1.2 mmol) according to
method B. The product was purified by preparative TLC (CH2Cl2);
yield: 0.260 g (73%). C22H19N3O2; MW: 357; m.p. 211–2138C (dec),
white powder. 1H NMR (DMSO-d6): 9.63 (bs, 1H, OH); 7.98
(t, J ¼ 1.6 Hz, 1H); 7.79–7.76 (m, 1H); 7.74–7.71 (m, 1H); 7.66–
ß 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
Triazole Derivatives as New Non-Steroidal Inhibitors of 17bHSD2
9
7.57 (m, 5H); 7.40 (t, J ¼ 8.0 Hz, 1H); 7.01–6.99 (m, 1H); 6.89
(d, J ¼ 8.6 Hz, 2H); 4.06 (s, 3H, NCH3); 3.83 (s, 3H, OCH3).
13C NMR (DMSO-d6): d 159.5, 157.5, 155.0, 140.8, 132.3, 130.0,
129.9, 129.4, 128.2, 128.0, 127.6, 126.6, 126.1, 118.1, 115.8, 115.0,
110.7, 55.1 (OCH3), 37.6 (NCH3). MS (ESI): 358 [MþH]þ.
(m, 1H); 7.19–7.18 (m, 1H); 6.97–6.95 (m, 1H); 6.91–6.89 (m, 1H);
4.14 (s, 3H, NCH3); 3.90 (s, 3H, OCH3); 3.87 (s, 3H, OCH3). 13C NMR
(CDCl3): d 160.9, 160.1, 159.8, 149.9, 147.5, 134.6, 132.0, 130.1,
129.6, 129.1, 128.3, 123.9, 118.9, 118.6, 115.9, 114.0, 111.7, 111.0,
55.4 (OCH3), 55.4 (OCH3), 37.3 (NCH3). MS (ESI): 378 [MþH]þ.
40-[3-(3-Methoxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
biphenyl-4-carbonitrile 8io
3-(3-Methoxyphenyl)-5-[5-(4-methoxyphenyl)-2-thienyl]-1-
methyl-1H-1,2,4-triazole 12b
The title compound was prepared by reaction of 5-(3-bromo-
phenyl)-3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g,
1.0 mmol) with 4-cyanobenzene boronic acid (0.176 g,
1.2 mmol) according to method B. The product was purified
by preparative TLC (CH2Cl2); yield: 0.256 g (69%). C23H18N4O;
MW: 366; m.p. 134–1368C (dec), white powder. 1H NMR
(CDCl3): 7.99 (t, J ¼ 1.5 Hz, 1H); 7.78–7.70 (m, 7H); 7.70
(t, J ¼ 1.4 Hz, 1H); 7.65 (t, J ¼ 7.7 Hz, 1H); 7.36 (t, J ¼ 8.0 Hz,
1H); 6.96 (m, 1H); 4.06 (s, 3H, NCH3); 3.89 (s, 3H, OCH3).
13C NMR (CDCl3): 161.2, 159.9, 155.1, 144.5, 140.1, 132.7, 132.1,
129.7, 129.6, 129.0, 128.9, 128.5, 127.9, 127.9, 118.8, 118.7, 115.9,
111.6, 111.0, 55.4 (NCH3), 37.1 (OCH3). MS (ESI): 367 [MþH]þ.
The title compound was prepared by reaction of 5-(5-bromo-
2-thienyl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole
(11)
(0.350 g, 1.0 mmol) with 4-methoxybenzene boronic acid
(0.114 g, 0.75 mmol) according to method A. The product was
purified by preparative TLC (CH2Cl2/MeOH, 10:0.25); yield:
0.237 g (63%). C21H19N3O2S; MW: 377; m.p. 87–898C (dec), white
powder. 1H NMR (CDCl3): d 7.76–7.74 (m, 1H); 7.70–7.69 (m, 1H);
7.59 (d, J ¼ 8.7 Hz, 2H); 7.52 (d, J ¼ 3.8 Hz, 1H); 7.36
(t, J ¼ 7.9 Hz, 1H); 7.26 (s, 1H); 6.97–695 (m, 3H); 4.14 (s, 3H,
NCH3); 3.91 (s, 3H, OCH3); 3.86 (s, 3H, OCH3). 13C NMR (CDCl3):
d 160.8, 159.9, 159.8, 150.0, 147.7, 132.0, 129.6, 129.3, 127.4,
127.1, 126.2, 122.7, 118.9, 115.9, 114.5, 111.0, 55.4 (OCH3),
55.4 (OCH3), 37.3 (NCH3). MS (ESI): 378 [MþH]þ.
5-(40-Fluoro-20-methylbiphenyl-4-yl)-3-(3-methoxyphenyl)-
1-methyl-1H-1,2,4-triazole 8ip
2-Methoxy-5-{5-[3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-
triazol-5-yl]-2-thienyl}pyridine 12c
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
4-fluoro-2-methylbenzene boronic acid (0.184 g, 1.2 mmol) accord-
ing to method B. The product was purified by preparative TLC
(CH2Cl2); yield: 0.278 g (74%). C23H20FN3O, MW: 373; m.p. 98–
1008C (dec), white powder. 1H NMR (CDCl3): 7.76–7.74 (m, 3H);
7.66 (t, J ¼ 1.5 Hz, 1H); 7.57 (t, J ¼ 7.5 Hz, 1H); 7.45–7.43 (m, 1H);
7.35 (t, J ¼ 8.0 Hz, 1H); 7.24–7.21 (m, 1H); 7.01–6.94 (m, 3H); 4.05
(s, 3H, NCH3); 3.89 (s, 3H, OCH3); 2.29 (s, 3H, CH3). 13C NMR (CDCl3):
163.2, 161.1, 159.9, 155.4, 141.8, 132.3, 131.2, 131.2, 131.0, 129.6,
128.7, 128.1, 117.3, 118.8, 117.0, 116.9, 115.9, 112.8, 112.7, 110.9,
55.4 (NCH3), 37.6 (OCH3), 20.6 (CH3). MS (ESI): 374 [MþH]þ.
The title compound was prepared by reaction of 5-(5-bromo-
2-thienyl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (11) (0.350 g,
1.0 mmol) with 2-methoxypyridine boronic acid (0.115 g,
0.75 mmol) according to method A. The product was purified by
preparative TLC (CH2Cl2/MeOH, 10:0.25); yield: 0.237 mg
(56%). C20H18N4O2S; MW: 378; m.p. 133–1358C (dec), white powder.
1H NMR (CDCl3): d 8.47 (d, J ¼ 2.3 Hz, 1H); 7.80 (m, 1H); 7.73–7.71
(m, 1H); 7.67–7.65 (m, 1H); 7.49 (d, J ¼ 3.8 Hz, 1H); 7.34 (t, J ¼ 7.9 Hz,
1H); 7.25 (d, J ¼ 2.3 Hz, 1H); 6.95–6.93 (m, 1H); 6.79 (d, J ¼ 8.5 Hz,
1H); 4.12 (s, 3H, NCH3); 3.97 (s, 3H, OCH3); 3.88 (s, 3H, OCH3). 13C NMR
(CDCl3): d 164.1, 161.0, 159.8, 149.8, 144.2, 144.0, 136.4, 132.0, 129.6,
129.1, 128.2, 123.5, 123.1, 118.9, 115.8, 111.3, 111.0, 55.4 (OCH3),
53.7 (OCH3), 37.3 (NCH3). MS (ESI): 379 [MþH]þ.
5-(30-Fluorobiphenyl-4-yl)-3-(3-methoxyphenyl)-1-methyl-
1H-1,2,4-triazole 8iq
Ether cleavage – general procedure for compounds 8a–h,
9, and 13a–c
The title compound was prepared by reaction of 5-(3-bromophenyl)-
3-(3-methoxyphenyl)-1H-1,2,4-triazole (7) (0.330 g, 1.0 mmol) with
3-fluorobenzene boronic acid (0.168 g, 1.2 mmol) according to
method B. The product was purified by preparative TLC (CH2Cl2);
To a solution of bis(methoxyphenyl) derivative (1 equiv) in dry
dichloromethane at room temperature, borontrifluoride
dimethyl sulfide complex (75 equiv) was added dropwise.
The reaction mixture was stirred for 20 h at rt. Water was added
to quench the reaction and the aqueous layer was extracted with
ethyl acetate. The combined organic layers were washed with
brine, dried over sodium sulfate, evaporated to dryness under
reduced pressure, and purified by preparative TLC.
1
yield: 0.254 g (70%), C22H18FN3O; MW: 359; oily product. H NMR
(CDCl3): 7.95 (s, 1H); 7.77–7.76 (d, J ¼ 7.6 Hz, 1H); 7.72–7.70 (m, 3H);
7.63–7.60 (m, 1H); 7.46–7.41 (m, 2H); 7.38–7.33 (m, 2H); 7.11–7.07 (m,
1H); 6.98–6.95 (m, 1H); 4.06 (s, 3H, NCH3); 3.90 (s, 3H, OCH3). 13C NMR
(CDCl3): d 161.2, 159.9, 155.3, 140.8, 132.3 130.5, 130.4, 129.6, 129.4,
128.8, 127.9, 127.7, 122.9, 118.9, 115.9, 114.8, 114.6, 114.2, 114.1,
110.9, 55.4 (OCH3), 37.0 (NCH3). MS (ESI): 360 [MþH]þ.
40-[3-(3-Hydroxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
3-(3-Methoxyphenyl)-5-[5-(3-methoxyphenyl)-2-thienyl]-1-
methyl-1H-1,2,4-triazole 12a
biphenyl-4-ol 8a
The title compound was prepared by reaction of 5-(40-methoxy-
biphenyl-4-yl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (8ia)
(0.100 g, 0.269 mmol) according to the general procedure
described above. The product was purified by preparative TLC
(CH2Cl2/MeOH, 10:0.25); yield: 41 mg (44%). C21H17N3O2; MW:
The title compound was prepared by reaction of 5-(5-bromo-
2-thienyl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole
(11)
(0.350 g, 1.0 mmol) with 3-methoxybenzene boronic acid
(0.114 g, 0.75 mmol) according to method A. The product was
purified by preparative TLC (CH2Cl2/MeOH, 10:0.25); yield:
0.248 g (66%). C21H19N3O2S; MW: 377; m.p. 109–1118C (dec),
white powder. 1H NMR (CDCl3): d 7.75–7.73 (m, 1H); 7.69–7.68
(m, 1H); 7.52 (d, J ¼ 3.8 Hz, 1H); 7.73–7.32 (m, 3H); 7.27–7.25
343. 1H NMR (DMSO-d6):
(d, J ¼ 8.3 Hz, 2H); 7.59 (d, J ¼ 8.5 Hz, 2H); 7.50–7.47 (m, 2H);
7.26 (t, J ¼ 7.8 Hz, 1H); 6.90 (d, J ¼ 8.5 Hz, 2H); 6.81–6.83 (m, 1H);
4.04 (s, 3H, CH3). 13C NMR (DMSO-d6): d 159.6, 157.8, 157.6, 154.5,
d
7.86 (d, J ¼ 8.3 Hz, 2H); 7.78
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10
Y. A. Al-Soud et al.
Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
141.6, 132.0, 129.7, 129.5, 129.0, 127.8, 126.0, 125.4, 116.4, 116.1,
115.9, 112.5, 37.1 (NCH3). MS (ESI): 344 [MþH]þ.
1.0 mmol) according to the general procedure described above.
The product was purified by preparative TLC (CH2Cl2/MeOH,
10:1); yield: 250 mg (76%). C20H15N3O2; MW: 329. 1H NMR
(CD3OD þ CD3COCD3, 2:1):
d
8.15 (d, J ¼ 8.4 Hz, 2H); 7.74
40-[3-(3-Hydroxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
(d, J ¼ 8.5 Hz, 2H); 7.57–7.55 (m, 2H); 7.33 (t, J ¼ 8.1 Hz, 1H);
7.28 (d, J ¼ 7.9 Hz, 1H); 7.17–7.12 (m, 2H); 6.93–6.91 (m, 1H);
6.83–6.81 (m, 1H). 13C NMR (CD3OD þ CD3COCD3, 2:1): d 160.5,
160.5, 159.3, 159.2, 143.8, 143.0, 131.6, 131.2, 131.2, 129.7, 128.4,
128.1, 119.3, 118.8, 118.1, 115.9, 114.8, 114.6. MS (ESI): 330
[MþH]þ.
biphenyl-3-ol 8b
The title compound was prepared by reaction of 5-(30-methoxy-
biphenyl-4-yl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (8ib)
(0.100 g, 0.269 mmol) according to the general procedure
described above. The product was purified by preparative TLC
(CH2Cl2/MeOH, 10:0.25); yield: 48 mg (52%). C21H17N3O2; MW:
343. 1H NMR (DMSO-d6): d 8.00–8.02 (m, 2H); 7.91–7.80 (m, 2H);
7.50–7.41 (m, 2H); 7.31–7.29 (m, 2H); 7.22–7.12 (m, 2H); 7.00–6.82
(m, 2H); 4.05 (s, 3H, CH3). 13C NMR (DMSO-d6): d 159.7, 157.9, 157.6,
154.4, 141.9, 140.5, 132.1, 130.1, 129.8, 129.1, 126.9, 126.6, 117.5,
116.5, 116.1, 115.0, 113.5, 112.5, 37.2 (NCH3). MS (ESI): 344 [MþH]þ.
40-[3-(4-Hydroxyphenyl)-1H-1,2,4-triazol-5-yl]biphenyl-
4-ol 8g
The title compound was prepared by reaction of 5-(40-meth-
oxybiphenyl-4-yl)-3-(4-methoxyphenyl)-1H-1,2,4-triazole
(8ig)
(0.357 g, 1.0 mmol) according to the general procedure
described above. The product was purified by preparative TLC
(CH2Cl2/MeOH, 10:1); yield: 243 mg (74%). C20H15N3O2; MW: 329.
1H NMR (CD3OD þ CD3COCD3, 2:1): d 8.13 (d, J ¼ 8.2 Hz, 2H);
7.95 (d, J ¼ 8.6 Hz, 2H); 7.69 (d, J ¼ 8.2 Hz, 2H); 7.55 (d, J ¼ 8.6 Hz,
2H); 6.92 (d, J ¼ 8.6 Hz, 2H); 6.89 (d, J ¼ 8.6 Hz, 2H). 13C NMR
(CD3OD þ CD3COCD3, 2:1): d 160.1, 160.1, 158.4, 142.9, 132.2,
132.1, 128.8, 128.8, 127.5, 127.2, 117.4, 116.5, 116.4. MS (ESI):
330 [MþH]þ.
40-[3-(4-Hydroxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
biphenyl-4-ol 8c
The title compound was prepared by reaction of 5-(40-methoxy-
biphenyl-4-yl)-3-(4-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (8ic)
(0.100 g, 0.269 mmol) according to the general procedure
described above. The product was purified by preparative TLC
(CH2Cl2/MeOH, 10:0.25); yield: 60 mg (64%). C21H17N3O2; MW:
343. 1H NMR (CD3OD): d 7.80 (d, J ¼ 8.6 Hz, 2H); 7.70–7.66
(m, 4H); 7.45 (d, J ¼ 8.5 Hz, 2H); 6.80 (d, J ¼ 8.5 Hz, 2H); 6.77
(d, J ¼ 8.5 Hz, 2H); 3.91 (s, 3H, CH3). 13C NMR (CD3OD): d 162.2,
160.1, 159.0, 156.9, 144.5, 132.3, 130.4, 129.2, 129.0, 127.8, 126.5,
123.2, 116.9, 116.5, 37.3 (NCH3). MS (ESI): 344 [MþH]þ.
40-[3-(4-Hydroxyphenyl)-1H-1,2,4-triazol-5-yl]biphenyl-3-
ol 8h
The title compound was prepared by reaction of 5-(30-meth-
oxybiphenyl-4-yl)-3-(4-methoxyphenyl)-1H-1,2,4-triazole
(8ih)
(0.357 g, 1.0 mmol) according to the general procedure
described above. The product was purified by preparative TLC
(CH2Cl2/MeOH, 10:1); yield: 214 mg (65%). C20H15N3O2; MW: 329.
1H NMR (CD3OD): d 8.11 (d, J ¼ 8.4 Hz, 2H); 7.89 (d, J ¼ 8.7 Hz,
2H); 7.71 (d, J ¼ 8.4 Hz, 2H); 7.27 (t, J ¼ 7.9 Hz, 1H); 7.15–7.10
(m, 2H); 6.92 (d, J ¼ 8.7 Hz, 2H); 6.81 (dd, J ¼ 2.4 Hz, 1H).
13C NMR (CD3OD): d 161.1, 160.0, 159.1, 143.8, 143.0, 131.0,
129.9, 129.3, 128.3, 128.0, 120.8, 119.3, 118.6, 116.9, 115.8,
114.8. MS (ESI): 330 [MþH]þ.
40-[3-(4-Hydroxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
biphenyl-3-ol 8d
The title compound was prepared by reaction of 5-(30-methoxy-
biphenyl-4-yl)-3-(4-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (8id)
(0.100 g, 0.269 mmol) according to the general procedure described
above. The product was purified by preparative TLC (CH2Cl2/MeOH,
10:0.25); yield: 63 mg (68%). C21H17N3O2; MW: 343. 1H NMR (CD3OD):
d 7.80 (d, J ¼ 8.7 Hz, 2H); 7.73–7.68 (m, 4H); 7.19 (t, J ¼ 7.9 Hz, 1H);
7.07–7.01 (m, 2H); 6.77 (d, J ¼ 7.0 Hz, 2H); 6.74–6.72 (m, 1H); 3.91
(s, 3H, CH3). 13C NMR (CD3OD): d 162.3, 160.1, 159.2, 156.7, 144.6,
142.6, 131.1, 130.4, 129.0, 128.4, 127.6, 123.2, 119.4, 116.5, 116.1,
114.9, 37.3 (NCH3). MS (ESI): 344 [MþH]þ.
30-[3-(3-Hydroxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
biphenyl-3-ol 9
The title compound was prepared by reaction of 5-(30-methoxy-
biphenyl-3-yl)-3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazole (8ii)
(0.372 g, 1.0 mmol) according to the general procedure described
above. The product was purified by preparative TLC (CH2Cl2/
MeOH, 10:1); yield: 233 mg (68%). C21H17N3O2; MW: 343;
1H NMR (CD3COCD3): d 8.53 (bs, 1H, OH); 8.42 (bs, 1H, OH); 8.09
(t, J ¼ 1.6 Hz, 1H); 7.86–7.97 (m, 2H); 7.68–7.63 (m, 3H); 7.33
(t, J ¼ 8.1 Hz, 1H); 7.29 (t, J ¼ 7.8 Hz, 1H); 7.23–7.21 (m, 1H);
6.91–6.88 (m, 2H); 4.12 (s, 3H, NCH3). 13C NMR (CD3COCD3):
d 162.3, 159.9, 159.5, 159.4, 157.0, 143.5, 143.5, 134.9, 132.0,
131.5, 131.2, 130.2, 129.4, 129.0, 120.2, 119.2, 117.8, 116.7,
115.8, 114.8, 38.7 (NCH3).
40-[3-(3-Hydroxyphenyl)-1H-1,2,4-triazol-5-yl]biphenyl-
4-ol 8e
The title compound was prepared by reaction of 5-(40-methoxy-
biphenyl-4-yl)-3-(3-methoxyphenyl)-1H-1,2,4-triazole (8ie) (0.357 g,
1.0 mmol) according to the general procedure described above.
The product was purified by preparative TLC (CH2Cl2/MeOH,
10:1); yield: 184 mg (56%). C20H15N3O2; MW: 329. 1H NMR
(CD3OD): d 8.07 (d, J ¼ 8.4 Hz, 2H); 7.68 (d, J ¼ 8.3 Hz, 2H);
7.53–7.51 (m, 4H); 6.80 (t, J ¼ 8.2 Hz, 1H); 6.92–6.87 (m, 3H).
13C NMR (CD3OD): d 160.8, 160.5, 159.2, 158.8, 143.9, 132.7,
131.6, 131.1, 129.1, 128.3, 128.0, 127.7, 118.8, 118.1, 116.9,
114.6. MS (ESI): 330 [MþH]þ.
3-{5-[3-(3-Hydroxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
2-thienyl}phenol 13a
The title compound was prepared by reaction of 3-(3-methoxy-
phenyl)-5-[5-(3-methoxyphenyl)-2-thienyl]-1-methyl-1H-1,2,4-triazole
(12a) (0.377 g, 1.0 mmol) according to the general procedure
described above. The product was purified by preparative TLC
40-[3-(3-Hydroxyphenyl)-1H-1,2,4-triazol-5-yl]biphenyl-
3-ol 8f
The title compound was prepared by reaction of 5-(30-methoxy-
biphenyl-4-yl)-3-(3-methoxyphenyl)-1H-1,2,4-triazole (8if) (0.357 g,
ß 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Arch. Pharm. Chem. Life Sci. 2012, 000, 1–12
Triazole Derivatives as New Non-Steroidal Inhibitors of 17bHSD2
11
(CH2Cl2/MeOH, 10:1); yield: 218 mg (62%). C19H15N3O2S; MW: 349.
1H NMR (CD3OD): d 7.45–7.39 (m, 3H); 7.28 (d, J ¼ 3.8 Hz, 1H); 7.16–
7.09 (m, 2H); 7.04–7.00 (m, 2H); 6.74 (m, 1H); 6.68 (m, 1H); 3.95
(s, 3H, CH3). 13C NMR (CD3OD): d 161.9, 159.2, 158.8, 151.3,
149.2, 135.8, 133.0, 131.3, 131.1, 130.8, 128.5, 125.1, 118.8,
118.3, 117.6, 116.7, 114.3, 113.7, 37.8 (NCH3). MS (ESI): 350 [MþH]þ.
were dissolved in acetonitrile. E1 and E2 were separated
using acetonitrile/water (45:55) as mobile phase in a C18 RP
chromatography column (Nucleodur C18 Gravity, 3 mm,
Macherey-Nagel) connected to a HPLC system (Agilent 1100
Series, Agilent Technologies, Waldbronn). Detection and
quantification of the steroids were performed using a radioflow
detector (Berthold Technologies, Bad Wildbad). The conversion
rate was calculated according to following equation:
%conversion ¼ (%E1/(%E1 þ %E2) ꢁ 100). Each value was calcu-
lated from at least three independent experiments.
3-{5-[5-(4-Hydroxyphenyl)-2-thienyl]-1-methyl-1H-1,2,4-
triazol-3-yl}phenol 13b
The title compound was prepared by reaction of 3-(3-methoxy-
phenyl)-5-[5-(4-methoxyphenyl)-2-thienyl]-1-methyl-1H-1,2,4-triazole
(12b) (0.377 g, 1.0 mmol) according to the general procedure
described above. The product was purified by preparative TLC
(CH2Cl2/MeOH, 10:1); yield: 189 mg (54%). C19H15N3O2S; MW:
349; white powder. 1H NMR (CD3OD): d 7.48 (d, J ¼ 3.9 Hz,
1H); 7.45–7.39 (m, 4H); 7.22 (d, J ¼ 3.9 Hz, 1H); 7.16
(t, J ¼ 7.9 Hz, 1H); 6.76–6.73 (m, 3H); 3.99 (s, 3H, CH3).
13C NMR (CD3OD): d 161.9, 159.4, 158.9, 151.5, 149.9, 133.0,
131.3, 130.8, 128.4, 127.1, 126.2, 123.6, 118.7, 117.5, 117.0,
114.3, 37.8 (NCH3). MS (ESI): 350 [MþH]þ.
Inhibition of 17bHSD1
The 17bHSD1 inhibition assay was performed similarly to the
17bHSD2 procedure. The cytosolic fraction was incubated with
NADH (500 mM), test compound and a mixture of unlabeled- and
[2,4,6,7-3H]-E1 (final concentration: 500 nM, 0.15 mCi) for
10 min. Further treatment of the samples and HPLC separation
was carried out as mentioned above.
We are grateful to the Deutsche Forschungsgemeinschaft (HA1315/8-1) for
financial support. We thank Beate Geiger and Jannine Ludwig for their
help in performing the enzyme inhibition tests (17bHSD1 and 17bHSD2).
Dr. Yaseen A. Al-Soud was grateful to the Alexander von Humboldt
Foundation (AvH) for a fellowship.
5-{5-[3-(3-Hydroxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-
2-thienyl}pyridin-2-ol 13c
The title compound was prepared by reaction of 2-methoxy-5-
{5-[3-(3-methoxyphenyl)-1-methyl-1H-1,2,4-triazol-5-yl]-2-thienyl}-
pyridine (12c) (0.378 g, 1.0 mmol) according to the general
procedure described above. The product was purified by
preparative TLC (CH2Cl2/MeOH, 10:1); yield: 216 mg
(62%). C18H14N4O2S; MW: 350; white powder. 1H NMR
(CD3OD þ CD3COCD3, 2:1): d 8.53 (d, J ¼ 2.0 Hz, 1H); 8.03
(m, 1H); 7.70–7.63 (m, 2H); 7.58–7.54 (m, 1H); 7.52
(d, J ¼ 3.9 Hz, 1H); 7.27 (t, J ¼ 8.1 Hz, 1H); 6.88–6.85 (m, 2H);
3.93 (s, 3H, CH3). 13C NMR (CD3OD þ CD3COCD3, 2:1): d 165.0,
161.2, 158.5, 150.4, 144.9, 144.3, 137.4, 133.2, 133.1, 132.7, 132.6,
130.5, 130.3, 125.0, 124.1, 118.1, 116.9, 113.7, 111.9, 37.8 (NCH3).
MS (ESI): 351 [MþH]þ.
The authors have declared no conflict of interest.
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Biological assays
[2,4,6,7-3H]-E2 and [2,4,6,7-3H]-E1 were bought from Perkin-
Elmer, Boston. Quickszint Flow 302 scintillator fluid was bought
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cipitation with ammonium sulfate from the cytosolic fraction.
´ ´
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